Discovery of pyrazole derivatives as potent inhibitor of NF-?B for possible benefit in abdominal aortic aneurysms

Article abstract of DOI:10.1007/s11696-021-01707-7

Numerous 3-(4-substitutedyphenyl)-5-methyl-1-phenyl-1H-pyrazole-4-carboxylate derivatives have been synthesized using H₂SO₄.SiO₂ as a catalyst. These derivatives were subsequently screened for NF-?B transcription inhibitor

Full text of DOI:10.1007/s11696-021-01707-7

Chemical Papers
https://doi.org/10.1007/s11696-021-01707-7
ORIGINAL PAPER
Discovery of pyrazole derivatives as potent inhibitor of NF‑ĸB
for possible beneft in abdominal aortic aneurysms
Yun Guo1 · Dongming Bao1 · Chaogang Zhang1
Received: 17 December 2020 / Accepted: 18 May 2021
© Institute of Chemistry, Slovak Academy of Sciences 2021
Abstract
Numerous 3-(4-substitutedyphenyl)-5-methyl-1-phenyl-1H-pyrazole-4-carboxylate derivatives have been synthesized using
H2SO4.SiO2 as a catalyst. These derivatives were subsequently screened for NF-ĸB transcription inhibitory activity in
RAW264.7 cells, where they showed mild to signifcant inhibition. Among the tested derivatives, Compound 4e was identifed
as a most potent NF-ĸB transcription inhibitor. The efect of compound 4e was further studied in abdominal aortic aneurysm
(AAA) animal model in BALB/c mice. The AAA was induced in mice by sub-cuteneous administration of Angiotensin-II
(Ang-II). Results suggest that compound 4e signifcantly inhibited infammation and oxidative stress in AAA mice as com-
pared to disease control. It also inhibits NF-ĸB and COX-2 in AAA mice as shown by western blot analysis. Collectively, it
was concluded that, compound 4e might acts as protective agent against AAA.
Keywords Pyrazole · One-pot synthesis · Silica · NF-ĸb · COX-2
Introduction:
arrangement of the aortic media. The human AAA tissue
showed higher expression of infammatory infltrates in both
the media and adventitia (Freestone et al. 1995; Shimizu
et al. 2006). Nuclear factor (NF)-kappaB (NF-ĸB), a tran-
scription factor responsible for controlling the action of vari-
ous genes linked with infammatory response in AAA (Saito
et al. 2013). It is found aberrantly activated in many experi-
mental studies of AAA to promote chronic infammation of
aortic walls, activation of MMPs (matrix metalloproteases)
and degradation of extra cellular matrix (ECM) (Rafetto and
Khalil 2008; Rabkin 2017).
Abdominal aortic aneurysm (AAA) is a life threatening
dwindling illness which often leads to death of the patient if
not treated well in time. The exact diagnosis of AAA is quite
cumbersome process due to its asymptomatic behaviour until
the rupture of aorta occurs. The AAA can be characterised
by the expansion of abdominal aortic diameter by 50% of
the original (Keisler and Carter 2015). The recent epide-
miological data suggests that men aged more than 65 year
are more prone to this illness and approximately 80% of the
mortality of this disease is due to rupture of aorta. The cur-
rent therapeutic modality to treat AAA is entirely based on
reconstructive surgery if the aortic diameter is over 5.5 cm
(Aggarwal et al. 2011; Setacci et al. 2016). However, there is
no medication for the patients with small AAA, as surgery is
fruitful only in major AAA. Thus, is worthwhile to discover
new agents for small AAA.
Pyrazole, a privileged heterocyclic scafold associ-
ated with numerous pharmacological activities ranging
from anti-infective agents to treat lifestyle diseases, such
as cardiovascular, diabetes (Faria et al. 2017). Various
studies have shown strong anti-infammatory activity of
pyrazole by inhibiting COX-2 and NF-ĸB in micro to
nano-molar range (Abdelgawad et al. 2017; Hassan et al.
2019),(Pippione et al. 2018; Masih et al. 2020b). The
clinical signifcance of pyrazole against infammation is
understood by the fact that, some of the best anti-infam-
matory agents are derived from pyrazole nucleus, such
as, Celecoxib (Gong et al. 2012), Lonazolac (Raulf and
König 1990) and Mepirizole (Boonyaratavej 1979; Tanaka
et al. 1989). However, the synthesis of substituted pyrazole
is quite tedious and difcult process due to involvement
Various studies have confrmed the role of infammation
in the development and progression of AAA. It is temporally
and spatially linked with disorder of the orderly lamellar
* Yun Guo
17358507362m@sina.cn
1
Department of Interventional, Qionglai Medical Center
Hospital, Qionglai 611530, Sichuan, China
Vol.:(0123456789)
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Chemical Papers
of multi-step reaction and associated purifcation, low
yield and complex workup. In this regard, various scien-
tist have developed facile synthesis of pyrazole to tackle
above-mentioned drawback by one-pot multi-step reac-
tion using numerous catalysts, such as, sodium ascorbate
(Kiyani and Bamdad 2018), molecular iodine (Mahdavi
et al. 2016), ionic liquid (Singh et al. 2013), nanoparticle
(Ghorbani-Vaghei et al. 2017), graphene oxide-phosphoric
acid (Zakeri et al. 2019). In a report by Jawale et al. they
have synthesized some pyrazole derivatives using SSA
as a catalyst in a facile and excellent yield (Jawale et al.
2011). Therefore, in the present study, we have employed
same methodology for the synthesis of numerous pyra-
zoles using SSA as a catalyst for potential beneft in AAA.
General procedure for the synthesis of substituted
pyrazole 4 (a‑g)
Aromatic aldehyde (1, 1 mmol), phenyl hydrazine (2, 1 mmol)
and ethyl acetoacetate (3, 1 mmol) was mixed together. To
this above mixture H2SO4–SiO2 (15 mol%) was added at
room temperature for the appropriate time as ascertained with
the help of TLC. The reaction mixture after the completion
of reaction was diluted with EtOAc (20 ml), fltered, water
(30 ml) added, the solution extracted with EtOAc (3×15 ml),
and the combined organic layers were dried over anhydrous
Na2SO4 and concentrated. The residue was subjected to col-
umn chromatography to obtain the pure desired product.
Ethyl 3‑(4‑hydroxyphenyl)‑5‑methyl‑1‑phenyl‑1H‑pyra‑
zole‑4‑carboxylate (4e)
Experimental
Yield: 89%; M.p: 176–178 °C; MW: 322.36; Rf (Benzene
and Ethyl acetate 1:1): 0.69; FTIR (νmax; cm−1 KBr): 3242
(O–H stretching), 3047 (Aromatic C-H stretching), 2945
(CH2 stretching), 2874 (CH3 stretching), 1716 (C=O stretch-
ing), 1635 (C=C stretching), 1573 (N–N stretching), 1462
(CH3 bending), 1405 (COO stretching), 1372 (CH2 bending),
1286 (C–O stretching), 1218 (C–N stretching), 784; 1H-NMR
(400 MHz, DMSO-d6, TMS) δ ppm: δ 1.23 (t, 3H, J=7.1 Hz,
CH3), 2.58 (s, 3H, CH3), 4.08 (q, 2H, J=7.1 Hz, CH2), 7.23
(d, 2H,, J=1.3 Hz, Ar–H), 7.41 (t, 1H, J=1.3 Hz, Ar–H),
7.58 (d, 2H, J=1.56 Hz, Ar–H), 7.62 (d, 2H, J=1.86 Hz,
Ar–H), 7.83 (d, 2H, J=1.3 Hz, Ar–H), 9.59 (s, 1H, Ar-OH);
13C-NMR (100 MHz, DMSO-d6, TMS) δ ppm: 162.5, 158.6,
153.7, 144.9, 133.8, 129.4, 128.7, 126.2, 125.7, 124.8, 116.3,
110.8, 60.9, 14.1, 11.6; Mass: 323.36 (M+H)+; Elemental
analysis for C19H18N2O3: Calculated: C, 70.79; H, 5.63; N,
8.69.Found: C, 70.76; H, 5.67; N, 8.73.
Chemistry
The chemicals used in the present study were obtained from
1
Sigma-Aldrich (USA). H NMR spectra were recorded in
DMSO-d6 on a Bruker Avance-400 NMR spectrometer
with TMS as the internal reference. 13C NMR spectra were
recorded on a Bruker Avance-100 NMR spectrometer in
DMSO-d6 on the same spectrometers with TMS as the inter-
nal reference. The multiplicity of a signal is indicated as: s
– singlet, d – doublet, t – triplet, q – quartet, m – multiplet,
br – broad, dd – doublet of doublets. Coupling constants (J)
are quoted in Hz and reported to the nearest 0.1 Hz. Infrared
spectra were recorded as a neat thin flm on a Perkin-Elmer
Spectrum One FT-IR spectrometer using Universal ATR
sampling accessories. Melting points were obtained using
MEL-TEMP (model 1001D). MS spectra were recorded on
an Agilent 1100 LC/MS. Elemental analysis was performed
with Vario elemental analyser and were in agreement with
the proposed structures within 0.4% of the theoretical
values.
Pharmacological activity
NF‑kB transcription inhibitory activity
The entire synthesized compound (at 10 µM) was tested for
their efect on the relative NF-ĸB transcriptional activity in
RAW264.7 cells using Dual‐Luciferase Reporter Assay Sys-
tem (Promega) as per the instruction provided by manufacturer.
Synthesis of H2SO4·SiO2
The synthesis of sulfuric acid adsorbed on silica gel was per-
formed with the earlier reported procedure, where silica gel
(29.5 g, 230–400 mesh size) was suspended in ethyl acetate
(60 mL) and then H2SO4 (1.5 g, 15.5 mmol, 0.8 mL of a 98%
aq. solution of H2SO4) was added to the above mixture. The
whole mixture was stirred magnetically for 30 min at rt and
after stirring the ethyl acetate was removed under reduced
pressure. The resulting mass was heated under vacuum for
72 h at 100–120 °C under vacuum to aford H2SO4–SiO2 as
a free fowing powder.
In vivo biological activity
Angiotensin II‑induced AAA mice model and treatment
The Male BALB/C male mice (n=18) after procuring from
the central animal house were kept in controlled tempera-
ture and humidity with alternate light and dark cycles mice.
They were fed with laboratory diet and water ad libitum. The
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Chemical Papers
following groups of mice were created for assessment of the
role of compound 4e.
the all-atom CHARMm (version c32b1) forcefeld with
Adopted Basis set Newton Raphson (ABNR) minimiza-
tion algorithm until the root mean square (r.m.s) gradi-
ent for potential energy was less than 0.05 kcal/mol/Å.
Using the ’Binding Site’ tool panel available in DS 3.5,
the minimized transcription factor NK-ĸB p50 homodi-
mer bound to a palindromic ĸB site was defned as recep-
tor, and the largest cavity was selected as a binding site for
docking runs. The centre of the sphere was created around
the identifed binding site, and side chains of the residues
in the binding site within the radius of the sphere were
assumed to be fexible during refnement of post-docking
poses.
Group 1: Control: Normal saline solution (0.5 mL, i.p.)
daily for 4 weeks (n=6).
Group 2: Angiotensin II-treated, (1,000 ng/kg/min angio-
tensin II, s.c.)+saline daily for 4 weeks (i.p.)
Group 3: Compound 4e (25 mg/kg) suspended in 0.5%
CMC i.p. daily for 4 weeks.
The mice was euthanized by an excess of anesthetic
(500 µl 3% pentobarbital sodium; Sigma-Aldrich).
Estimation of angiotensin II‑induced AAA mice
The periadventitial tissue from the aortic wall was removed
after fxing aorta 4% cold paraformaldehyde by using dis-
secting microscope. The aorta was macroscopically exam-
ined to record maximal external diameter of the suprarenal
aorta using MultiGuage 3000 imaging processing software.
The state of aortic aneurysm was achieved when the aortic
outer diameter was found enlarged>50%.
Ligand setup Using the built-and-edit module of DS 3.5,
the compound 4e was built, all-atoms CHARMm forcefeld
parameterization assigned and then minimized using the
ABNR method. A conformational search of the ligand was
carried out using a stimulated annealing molecular dynam-
ics (MD) approach. The ligand was heated to a temperature
of 700 K and then annealed to 200 K. Thirty such cycles
were carried out. The transformation obtained at the end of
each cycle was further subjected to local energy minimiza-
tion, using the ABNR method. The 30 energy-minimized
structures were then superimposed and the lowest energy
conformation occurring in the major cluster was taken to be
the most probable conformation.
Measurement of infammation
TNF-α, IL-1β and IL-6 serum levels were determined using
the eBioscience mouse ELISA antibody set. Absorbance was
determined at 450 nm wavelength using an ELISA reader
(Spectramax Plus, Molecular Devices, LLC, Sunnyvale, CA,
USA).
Western blot analysis of NF‑κB and COX‑2
Docking and scoring
The isolated protein from the whole aorta was probed to 10%
SDS-PAGE for 50 min and transferred to PVDF membrane.
The membrane was blocked with 5% defat milk in tris-buf-
ered saline with 0.1% Tween 20 for 1 h at 4 °C and then
incubated with rabbit anti-mouse NF-κB, COX-2 (1:1,000),
and β-actin (1:2,000) primary antibodies at 4 °C for over-
night. Afterwards, the membrane was incubated with anti-
mouse IgG HRP-conjugated secondary antibodies (1:5000).
Protein expression was detected with an enhanced chemilu-
minescence detection kit.
CDOCKER protocol of DS 2.5 was used for the docking of
compound 4e with transcription factor NK-ĸB p50 homodi-
mer bound to a palindromic ĸB site. The receptor protein
conformation was kept fxed during docking, and the docked
poses were further minimized using all-atom CHARMm
(version c32b1) forcefeld and smart minimization method
(steepest descent followed by conjugate gradient) until r.m.s
gradient for potential energy was less than 0.05 kcal/mol/Å.
The atoms of ligand and the side chains of the residues of the
receptor within 5 Å from the centre of the binding site were
kept fexible during minimization. Finally, the best docked
pose of compound 4e was analysed for it interaction with
receptor protein using the Visualizer of the software.
Docking
The 3D X-ray crystal structure of transcription factor NK-
kappa B p50 homodimer bound to a palindromic kappa B
site was used as a target protein model for this study (1nfk.
pdb). All computational analyses were carried out using Dis-
covery Studio 3.5.
Statistical analysis
Data are presented as means standard error Statistical
analysis was performed using Student’s t-test for paired or
unpaired data, and data were assessed using Graphpad Prism
(version 5). P<0.05 was considered to indicate a statistically
signifcant diference.
Preparation of receptor The water was removed from the
protein, and the bond orders were corrected. The hydro-
gen atoms were added, their positions optimized using
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Chemical Papers
Scheme 1 Synthesis of pyra-
zole derivatives 4 (a-g)
Table 1 Efect of catalyst loading on the reaction
Table 2 Efect of Temperature on the reaction condition
Entry
H2SO4·SiO2 (in
mol %)
Time (in min)
Yield (in %)b
Entry
Temperature (in C°)
Time (in min)
Yield (in %)b
1
2
3
4
5
10
1260
31
20
11
6
23
92
62
40
32
1
2
3
4
5
10
15
20
650
265
31
30
47
92
60
Room temperature
50
100
140
19
aReaction conditions: Benzaldehyde (1 mmol), phenylhydrazine
(1 mmol), and ethyl acetoacetate (1 mmol); catalyst, room tempera-
ture, solvent free
aReaction conditions: Benzaldehyde (1 mmol), phenylhydrazine
(1 mmol), and ethyl acetoacetate (1 mmol); H2SO4·SiO2 (15 mol %),
solvent free
bIsolated and unoptimized yield
bIsolated and unoptimized yield
Results and discussion
nearly 30 min to complete. In the next runs (Table 2, entry
3, 4 and 5), it was marked to note that reaction took shorter
time to aford the product upon increasing the temperature,
however, it does not found favourable to the product yield,
which found signifcantly reduced as the temperatures
rises. Thus, reaction conducted at room temperature seems
favourable as compared to the other temperature condi-
tions. Recyclability/reusability of catalyst is a vital point
which needs to be considered for the applicability of the
catalyst for any reaction. The reusability of a catalyst has
a signifcant impact in terms, limiting waste generation,
economically viable, and environment friendly. Thus, in
the next instance, the recyclability of the catalyst was then
evaluated. The catalyst was recovered by fltration, washed
with ethyl acetate (10 mL), dried and reused for a further
catalyzing the reaction. As shown in Table 3, the catalyst
can be used for six-consecutive runs without much loss
of ability. However, the signifcant reduction in product
yield was obtained in seventh-run of catalyst reusability.
Thus, it could be suggested that catalyst can be recovered
and reused for 6 cycles at least, fully preserving its activity
and selectivity. On the basis of the above observation, it
could be suggested that, 15 mol % loading of the catalyst
at a room temperature was efcient to catalyze the reaction
with maximum yield.
Synthesis of Target molecules 4 (a‑j)
The synthesis of pyrazole derivatives has been achieved
using catalyst (H2SO4.SiO2, silica-supported sulfuric
acid), Scheme 1. Initially, the optimal catalyst loading of
silica-supported sulfuric acid catalyst for the reaction was
identifed using benzaldehyde, phenylhydrazine and ethy-
lacetoacetate in equi-molar quantity as a model reaction.
As shown in Table 1, at the initial catalytic load of 5 mol
%, the product yield was found lowest, the product conver-
sion being only 30% in approximately 11 h, the longest
reaction time (Table 1, entry 1). The twofold increment of
the catalytic load to 10% (Table 1, entry 2) showed marked
improvement in the reaction time with marginal increment
in the product yield. The highest conversion of the product
was observed at 15 mol % catalyst loading with subsequent
reduction in the reaction time (Table 1, entry 3). A further
increase in catalyst load (20 mol %) seems to hamper the
reaction as suggested by signifcant reduction in the prod-
uct yield, albeit it improves the reaction time (Table 4).
Thus, based on the above observation, 15 mol % is identi-
fed as an optimal catalyst load for the efcient synthesis of
pyrazole. The reaction conditions were further optimized
studying the efect of the temperature on the catalytic ef-
ciency and product yield using the same model reaction
template. As shown in Table 2, at 10 °C, the reaction took
maximum time to complete with the least product yield.
However, the maximum product yield was obtained at the
reaction conducted at the room temperature which took
Finally, with optimized conditions in our hands, we
checked the scope of this catalytic methodology for the
synthesis of substituted pyrazole using numerous electron-
donating and withdrawing substituents as a starting mate-
rial. As shown in Table 4, the compounds containing elec-
tron-withdrawing group were obtained in excellent yield as
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Table 3 Recyclability of
Table 5 Efect of compound 4 (a-g) on NF-ĸB transcriptional activ-
Entry
Yield (in %)b
Run
catalyst for reaction
ity in LPS-stimulated RAW264.7 cells
1
2
3
4
5
6
92
92
92
91
90
89
68
1
2
3
4
5
6
7
Compound
Substituent
Relative NF-ĸB transcrip-
tional activity (NF-ĸB/TK,
fold)a
4a
H
4-Cl
4-F
4-Br
4-OH
4-OCH3
4-CH3
5.03 1.05#
3.76 1.32#
4.87 0.97#
4.80 0.69#
1.34 0.47#
2.45 0.36#
3.02 1.02#
1.82 0.21
0.39 0.07
5.67 0.95**
4b
4c
4d
aReaction conditions: Benzal-
dehyde (1 mmol), phenylhydra-
zine (1 mmol), and ethyl ace-
toacetate (1 mmol); H2SO4·SiO2
(15 mol %), room temperature,
solvent free
4e
4f
4 g
Celecoxib
Control
LPS
bIsolated and unoptimized yield
LPS, lipopolysaccharide; NF-ĸB, nuclear factor kappa B
N=3
compared to their electron donating counterparts. Moreover,
compound with no-substitution showed highest yield among
the synthesized derivatives. The formation of pyrazoles was
established on the basis of a melting point which was found
in the agreement with earlier reported literature (Meng et al.
2016).
aAt 10 µM
#P<.01 versus LPS
**P<.01 versus control
inhibitory activity was further found reduced upon intro-
duction of methoxy (4f) and methyl (4g). On close inspec-
tion of comparative inhibitory activity of tested derivatives,
compound 4e found highly active among the tested deriva-
tives showing approximately similar inhibitory activity to
Celecoxib as a standard.
NF‑ĸB Transcriptional inhibition activity
The synthesized pyrazole derivative 4 (a-g) were evaluated
for relative NF-ĸB transcriptional activity in LPS stimu-
lated RAW264.7 cells. As shown in Table 5, entire set of
compounds displayed signifcant to moderate NF-ĸB tran-
scriptional inhibitory activity. The compound 4a showed
least NF-ĸB transcriptional inhibitory activity. The inhibi-
tory activity was considerably improved on the introduc-
tion chloro group (4b). The replacement of chloro with the
fuoro (4c) showed reduced activity. However, no signif-
cant change in activity was reported by the introduction of
bromo (4d) in place of fuoro. The activity was signifcantly
increased upon the insertion of hydroxy (4e) which render
compound highly potent among the tested derivatives. The
Pharmacological activity of Compound 4e
against abdominal aortic aneurysm (AAA) in mice
Impressed by the potent NF- ĸB transcriptional efect of
compound 4e, in the next part, we aimed to investigate
whether compound 4e has any efect on abdominal aortic
aneurysm in the rats. The AAA in mice was induced by
administration of angiotensin II (Ang-II). Various studies
have shown a clear link between Ang-II and AAA, where
Table 4 Synthesis of pyrazole
Entry
Substituent
Yield (in %)
M.P. (in °C, found)
M.P. (in °C, reported)
(Meng et al. 2016)
derivatives 4 (a-j)
4a
4b
4c
4d
4e
4f
H
4-Cl
4-F
4-Br
4-OH
4-OCH3
4-CH3
92
90
90
91
86
85
87
142–144
175–178
109–112
175–178
176–178
124–128
172–175
143–144
176–177
110–111
178–179
178–179
124–125
175–176
4 g
aReaction conditions: Substituted Benzaldehyde (1 mmol), phenylhydrazine (1 mmol), and ethyl acetoac-
etate (1 mmol); H2SO4·SiO2 (15 mol%), room temperature, solvent free
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it causes induction VCAM-1 expression by activating
NF-κB–dependent gene expression. In addition, Ang-II
stimulates production of reactive oxygen species (ROS) by
inducing a vascular NADH oxidase (Daugherty et al. 2000;
Golledge et al. 2006; Weintraub 2009; Cao et al. 2010). As
shown in Fig. 1, the mice treated with Ang-II showed higher
incidence of AAA in mice together with high mortality and
max aortic aneurysm as compared to control. However, upon
administration of compound 4e (25 mg/kg) showed signif-
cant reduction in these indices as compared with Ang-II
model group. The efect of compound 4e was then inves-
tigated on the level of various pro-infammatory cytokines
(TNF-α, IL-1β and IL-6) in AAA mice. As shown in Fig. 2,
the level of these cytokines was found elevated in AAA-
model group as compared to control, whereas, their level
were found signifcantly reduced in compound 4e treated
mice. Oxidative stress is the characteristic hallmark of AAA
(McCormick et al. 2007; Kuivaniemi et al. 2015). Thus,
the efect of compound 4e was investigated on the level of
various biomarkers of oxidative stress in AAA mice, such as
MDA, SOD and GSH. It has been found that compound 4e
causes signifcant reduction in MDA content and increases
SOD and GSH activity (Fig. 3). Various studies showed
excellent correlation between the higher expressions NF-ĸB
and COX-2 and incidence of AAA (Guo et al. 2006). Thus,
the efect of compound 4e was further investigated on the
expression of NF-ĸB and COX-2 using western blot analy-
sis. As shown in Fig. 4, compound 4e signifcantly reduces
the expression of both NF-ĸB and COX-2 in AAA rats.
Molecular docking analysis of compound 4e with NF‑ĸB
Now days, molecular docking has attracted wide atten-
tion from scientists across the globe working on drug dis-
covery. It is a powerful tool that can able to predict the
3D-alignment of ligand into the active site of receptor of
interest which allows us to illustrate the action of small
molecules into the binding site of aimed proteins as well
Fig. 1 Efect of compound 4e (25 mg/kg) on the indices of abdominal
aortic aneurysms in rats induced by Angiotension-II. a incidence of
abdominal aortic aneurysms, b max. aortic aneurysms, and c mortal-
ity rate. ##P<0.01 vs. the control group and **P<0.01 vs. the angio-
tensin II group. Data are presented as means SEM
Fig. 2 Efect of compound 4e (25 mg/kg) on various pro-infam-
matory cytokines in abdominal aortic aneurysms in rats induced by
Angiotension-II. a TNF-α, b IL-1β, and c IL-6. ##P<0.01 vs. the con-
trol group and **P<0.01 versus the angiotensin II group. Data are
presented as means SEM
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Fig. 3 Efect of compound 4e (25 mg/kg) on various indices of oxi-
dative stress in abdominal aortic aneurysms in rats induced by Angi-
otension-II. a MDA, b SOD, and c GPx. ##P<0.01 vs. the control
group and **P<0.01 versus the angiotensin II group. Data are pre-
sented as means SEM
Fig. 4 Efect of compound
4e (25 mg/kg) on various
pro-infammatory cytokines in
abdominal aortic aneurysms in
rats induced by Angiotension-
II. a TNF-α, b IL-1β, and c
IL-6. ##P<0.01 vs. the control
group and **P<0.01 versus the
angiotensin II group. Data are
presented as means SEM
as to explicate elementary biochemical processes (Meng
et al. 2012; Sliwoski et al. 2014). Therefore, encouraged
by the excellent inhibitory activity of compound 4e in the
both in-vitro and in-vivo experiments, we envisage to per-
form molecular docking experiment of 4e with 3D-crystal
structure of NF-ĸB. Results of the docking study presented
in Figs. 5 and 6. It has been found that compound 4e cre-
ated numerous interactions into the active site of NF-ĸB
by creating inter-atomic contacts with neighbouring amino
acid residues (shown in color coded form in Fig. 6). The
oxygen atom of ketone and hydroxy group and pyrazole
Nitrogen atom of compound 4e showed formation of van
der Waals and Hydrogen bond with Lys145 of Chain A and
B and Thr143 of Chain B. Moreover, the methyl of com-
pound 4e interacted with Cys59 of Chain A of receptor via
alkyl bond. Two pi-alkyl interactions was reported by phe-
nyl linked to Nitrogen atom of pyrazole with Lys146 and
Val58 of Chain B of the NF-ĸB. The interaction shown by
compound 4e found similar with earlier reported results
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Conclusion
Our study demonstrated the usefulness of synthesized
pyrazole. The compounds found excellent inhibitor of
NF-ĸB and showed protective efect against aortic aneu-
rysms via inhibition of oxidative stress and infammation.
However, much studies are need on these derivative make
them viable lead for clinical use.
Declarations
Conflict of interest The authors declare that they have no confict of
interest.
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