Design, synthesis, biological evaluation, and molecular docking study on triazine based derivatives as anti-inflammatory agents

Article abstract of DOI:10.1016/j.molstruc.2021.130760

In an attempt to develop new anti-inflammatory agents, design, synthesis, pharmacological activities, and docking study of two groups of triazine-based derivatives were reported. Nine compounds (5a-5d and 10a-10e) consisting of triazine, vanillin, and phenylpyrazole were synthesized through the pharmacophore hybridization method. After confirmation of the structure of the synthesized compounds using spectroscopic methods (FT-IR, and NMR spectral data), their anti-inflammatory activity was evaluated using carrageenan-induced paw edema model in male Wistar rats (200–220 g) administered intraperitoneally at doses of 100 and 200 mg/kg. A group of rats received indomethacin (10 mg/kg) as the standard drug. Among compounds 5a to 5d, only compounds 5c and 5d showed a significant anti-inflammatory effect (p < 0.01). Also compound 10a at a dose of (200 mg/kg) and compounds 10b, 10c, 10d and 10e at both doses showed significant anti-inflammatory activity and this effect for 10a (200 mg/kg) and both doses of 10b and 10e was comparable with indomethacin. While indomethacin reduced paw edema by 90%, 10b as the most potent tested compound reduced edema by 93%. The synthesized compounds were docked into the binding sites of both cyclooxygenase-1- and 2- isoenzymes (COX-1 and COX-2) to explore their binding mode and possible interactions of these ligands.

Full text of DOI:10.1016/j.molstruc.2021.130760

Journal of Molecular Structure 1243 (2021) 130760
Contents lists available at ScienceDirect
Journal of Molecular Structure
journal homepage: www.elsevier.com/locate/molstr
Revised
Design, synthesis, biological evaluation, and molecular docking study
on triazine based derivatives as anti-inflammatory agents
Parvin Asadia,b, Mohsen Alvania, Valiollah Hajhashemib,c, Mahboubeh Rostamia,b
,
Ghadamali Khodarahmia,b,∗
a Department of Medicinal Chemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, 81746-73461, Iran
b Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Science, Isfahan University of Medical Sciences, Isfahan, Iran
c Department of Pharmacology and Toxicology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran
a r t i c l e i n f o
a b s t r a c t
Article history:
In an attempt to develop new anti-inflammatory agents, design, synthesis, pharmacological activities,
and docking study of two groups of triazine-based derivatives were reported. Nine compounds (5a-5d
and 10a-10e) consisting of triazine, vanillin, and phenylpyrazole were synthesized through the pharma-
cophore hybridization method. After confirmation of the structure of the synthesized compounds using
spectroscopic methods (FT-IR, and NMR spectral data), their anti-inflammatory activity was evaluated
using carrageenan-induced paw edema model in male Wistar rats (200–220 g) administered intraperi-
toneally at doses of 100 and 200 mg/kg. A group of rats received indomethacin (10 mg/kg) as the standard
drug. Among compounds 5a to 5d, only compounds 5c and 5d showed a significant anti-inflammatory ef-
fect (p < 0.01). Also compound 10a at a dose of (200 mg/kg) and compounds 10b, 10c, 10d and 10e at
both doses showed significant anti-inflammatory activity and this effect for 10a (200 mg/kg) and both
doses of 10b and 10e was comparable with indomethacin. While indomethacin reduced paw edema by
90%, 10b as the most potent tested compound reduced edema by 93%. The synthesized compounds were
docked into the binding sites of both cyclooxygenase-1- and 2- isoenzymes (COX-1 and COX-2) to explore
their binding mode and possible interactions of these ligands.
Received 6 April 2021
Revised 19 May 2021
Accepted 21 May 2021
Available online 28 May 2021
Keywords:
Triazine derivatives
Anti-inflammatory agent
Docking
Cyclooxygenase-2 isoenzymes
© 2021 Elsevier B.V. All rights reserved.
1. Introduction
inflammatory agents with a potential for clinical use, associating
with minimal or even without any adverse effects [9-12].
Non-steroidal anti-inflammatory drugs (NSAIDs) are the most
widely used medicines for the treatment of different types of
inflammations which exert their action by inhibition of cy-
clooxygenase enzymes [1,2]. Traditional NSAIDs, inhibit the COX-
1 isoenzyme which is mainly responsible for the gastrointesti-
nal side effects and/or inhibits both COX-1 and COX-2 isoenzymes
[3,4]. Inhibition of the COX-2 isoenzyme leads to a reduction of
prostaglandins and thromboxanes which are responsible for in-
flammatory effects [5,6]. Therefore, to have an anti-inflammatory
drug without any gastrointestinal side effects, there is a contin-
ual need for a selective COX-2 inhibitor. Drugs such as celecoxib
(Scheme 1), valdecoxib, and rofecoxib are the most important mar-
keted selective COX-2 inhibitor drugs [7,8]. But their administra-
tion is associated with some myocardial thrombotic side effects.
So it is an important goal in medicinal chemistry to find new anti-
Merging two or more pharmacophores into a single molecule
is an approach, frequently used in drug design and discovery that
has recently been widely used to introduce new anti-inflammatory
agents with potent activity and diminished side effects [13-15].
Triazines, as analog to the benzene ring, are heterocyclic scaf-
folds used in diverse compounds with
a variety of biological
activities such as analgesic, anti-tuberculosis, anti-fungal, anti-
cancer, anti-protozoal, antimalarial, anti-viral, anti-microbial, and
anti-inflammatory activity [16,17]. Due to their versatile structure
(three isoforms depending on the positions of the nitrogen atoms)
and also the different derivatives that can be synthesized from
them, this heterocyclic ring has been increasingly studied in recent
years. According to a review article published in 2019, about 35.17%
of evaluated triazines were revealed promising anti-inflammatory
properties [18]. So in designing new anti-inflammatory agents, the
triazine ring could be a proper candidate. Additionally, vanillin
and phenylpyrazole derivatives also represent important classes of
compounds due to their highly pronounced pharmacological and
biological activities such as anti-inflammatory effects [19-21].
∗
Corresponding author.
E-mail address: khodarahmi@pharm.mui.ac.ir (G. Khodarahmi).
https://doi.org/10.1016/j.molstruc.2021.130760
0022-2860/© 2021 Elsevier B.V. All rights reserved.

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
Scheme 1. Structures of vanillin (I), celecoxib (II), the anti-inflammatory derivative of triazine (III), and designed vanillin-triazine derivatives (IV) and phenylpyrazole-triazine
derivatives (V).
The present work is a part of our ongoing research program
aiming at developing a variety of heterocyclic systems for bio-
logical and pharmacological evaluation. Considering celecoxib as
a selective COX-II inhibitor (Scheme 1) which is consisted of two
aryl moieties attached to a central heteroaryl (pyrazole) ring, two
groups of compounds were designed and synthesized. To design
the target compounds, vanillin and triazine moieties were selected
since the anti-inflammatory properties of vanillin as a natural sub-
stance have already been proven, and also this fact that triazine
derivatives are key structures for the development of the new
chemical class of anti-inflammatory agents [22,23]. In series 1, the
central pyrazole ring of the celecoxib was replaced by the tri-
azine ring, and one of the aryl rings was exchanged with vanillin
(Scheme 1). In the other series, the triazine ring was placed instead
of one of the two aryl moieties and binds to phenylpyrazole, which
is present in the structure of celecoxib (Scheme 1). The structures
of all synthesized compounds were characterized by their melting
points, FT-IR, and NMR spectral data. The anti-inflammatory activ-
ity of the newly synthesized compounds was evaluated using the
rat carrageenan-induced foot paw edema model [24], with the use
of indomethacin as comparable reference drugs or positive control.
Finally, synthesized compounds were docked into the binding sites
of both COX-1 and COX-2 to explore their complementarity with
the specified binding pocket.
2. Experimental
2.1. Material and method
Melting points were determined on an electrothermal 9200
melting point instrument (UK) and are uncorrected. Spectroscopic
methods were applied for the characterization of the synthesized
compounds. Infrared (IR) spectra were recorded as KBr plates using
a Jasco-680 FT-IR spectrometer (Japan). 1H NMR spectra were mea-
sured on an NMR Spectrophotometer (Bruker 500 MHz, Germany)
with TMS as the internal standard, where J (coupling constant) val-
ues are estimated in Hertz (Hz). Thin-layer chromatography tech-
niques with Merck silica gel 60 F254 plates (Germany) were used
for evaluation of reaction progression and checking the purity of
synthesized compounds.
2

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
Scheme 2. Synthesis of vanillin-triazine and phenylpyrazole-triazine derivatives.
2.2. Synthesis of vanillin-triazine and phenylpyrazole-triazine
derivatives
mixture at 0–5 °C. The resulting mixture was then stirred for a
further 1 hour at the same temperature (0–5 °C) and finally, the
solid product was filtered, washed with water, and dried under
vacuum to obtain 4,6-dichloro-1,3,5-triazine-2-ylphenylamine
derivatives as white solids.
The designed target compounds were synthesized according to
scheme 2.
2.2.1. General procedure for the synthesis of
2.2.2. General procedure for the synthesis of vanillin-triazine
compounds 5a-5f
4,6-dichloro-1,3,5-triazine-2-ylphenylamine derivatives (3a-3d)
4,6-Dichloro-1,3,5-triazin-2-ylphenylamine derivatives were
prepared following the earlier reported procedures [25]. A solution
of cyanuric chloride (9.22 g, 0.05 mol) in 25 mL of acetone was
placed in an ice-water bath to keep the temperature between 0
and 5 °C. Then 0.05 mol of different amines were added slowly
and after 30 min 7.2 g (0.05 mol) of NaHCO3 was added to the
In this series of compounds, the second chlorine group of the
cyanuric chloride was replaced by vanillin. To a solution of syn-
thesized compounds (3a-3d) (25.5 mmol) in 15 mL of DMF, 3.8 g
(25.5 mmol) of vanillin, and 3.5 g (25.5 mmol) potassium carbon-
ate were added. The reaction mixture was then stirred at 60 °C
for 12 h. The progress of the reaction was monitored by TLC. Upon
3

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
completion, the mixture was decanted in crushed ice; the sepa-
rated solid was collected and recrystallized from ethanol.
(15 g in 10–15 mL of distilled water) was added to furnish a yel-
low solid which was filtered off and washed with ice-cold water.
The obtained solid dissolved in a warm solution of NaOH (6 g in
100 mL of H2O) at 50 °C and then the pH of the solution was ad-
justed to pH = 5 by dropwise addition of 10% HCl. In the next step
hydrazine hydrate (4 mL, 0.13 mol) was added dropwise and the
solution was stirred overnight. Finally, the resulting 4-phenyl-1H-
pyrazole was filtered off as a white crystal and dried.
2.2.2.1. 4-(4–chloro-6-(phenylamino)−1,3,5-triazin-2-yloxy)−2-
methoxybenzaldehyde (5a). C17 H13ClN4O3; Yield: 83%, White
Solid, m.p. 145–146 °C, IR (KBr, Cm−1), 3362 (N H, st), 3062,
–
–
–
–
(C H, aromatic, st), 2932 (C H, aliphatic, st), 2742 (C H, aldehyde,
st), 1714 (C = O, aldehyde, st), 1557 (C = N), 1415–1612 (C = C
1
–
aromatic), 768 (C Cl). H NMR: (400 MHz; DMSO: d6): 11.2 (1H, s,
CHO), 8.0 (1H, d, ArH, j = 8), 7.7 (1H, S, ArH),δ 7.4 (2H, d, ArH,
j = 6.8), 7.2 (4H, m, ArH), 6.5 (1H, NH, br), 3.72 (3H, s, OCH3).
2.2.5. General procedure for the synthesis of phenylpyrazole-triazine
derivatives (10a-10e)
For the synthesis of phenylpyrazole-triazine derivatives 10a-10e,
compounds 8a-8d (20 mmol) were dissolved in DMF (20 mL) and
then phenyl pyrazole (2.88 g, 20 mmol) was added and the so-
lution was stirred at 60 °C for 3 h in presence of K2CO3 (2.7 g,
20 mmol). After completion of the reaction by monitoring with
TLC, the reaction mixture was poured in water and the result-
ing precipitate was filtered and dried. As another derivative of
this class, phenylpyrazole (2.88 g, 20 mmol) was reacted with an
equimolar ratio with cyanuric chloride (3.70 g, 20 mmol) in ace-
tone (15 ml) at room temperature. The product, which was precip-
itated in the mixture, was used for further investigations. The ob-
tained products were recrystallized in ethyl acetate and n-heptane.
2.2.2.2. 4-(4-(4-bromophenylamino)−6–chloro-1,3,5-triazin-2-
yloxy)−2-methoxybenzaldehyde
(5b). C17 H12BrClN4O3;
86%, Yellow Solid, m.p. 140–142 °C, IR (KBr, Cm−1), 3350 (N H, st),
Yield:
–
–
–
3132, (C H, aromatic, st), 2824 (C H, aldehyde, st), 1711 (C = O,
aldehyde, st), 1651 (C = N), 1391–1680 (C = C aromatic), 1259
(C O, st), 749 (C Cl, st). 1H NMR: (400 MHz; DMSO: d6): 11.12
(1H, s, CHO), 8.1 (1H, d, ArH, j = 8), 7.7(3H, m, ArH), 7.42 (2H, d,
ArH, j = 7), 7.2 (1H, d, ArH, j = 8), 6.7 (1H, NH, br), 3.69 (3H, s,
OCH3).
–
–
2.2.2.3. 4-(4-(4-formyl-3-methoxyphenoxy)−6–chloro-1,3,5-triazin-
2-ylamino)benzonitrile (5c). C18 H12ClN5O3; Yield: 83%, White
Solid, m.p. 150–152 °C, IR (KBr, Cm−1), 3270 (N H, st), 3087,
2.2.5.1. 4–chloro-N-methyl-6-(4-phenyl-1H-pyrazol-1-yl)−1,3,5-
–
triazin-2-amine (10a). C13H11 ClN6; Yield: 61%, White Solid, m.p.
–
–
–
(C H, aromatic, st), 2925 (C H, aliphatic, st) 2784 (C H, aldehyde,
110–111 °C, IR (KBr, Cm−1), 3221 (N H, st), 3091 (C H, aromatic,
–
–
≡
st), 2259 (C N, st), 1717 (C = O, aldehyde, st), 1571 (C = N),
1400–1600 (C = C aromatic), 1310 (C O, st), 750 (C Cl, st). 1H
NMR: (400 MHz; DMSO: d6): 11.18 (1H, s, CHO), 8.1 (1H, d, ArH,
j = 8), 7.7 (3H, m, ArH), δ 7.36 (2H, d, ArH, j = 6.8), 7.2 (1H, d,
ArH, j = 8), 6.4 (1H, NH, br), 3.70 (3H, s, OCH3).
–
st), 2921 (C H, aliphatic, st), 1552 (C = N), 1400–1600 (C = C
–
–
aromatic), 770 (C Cl, st). 1H NMR: (400 MHz; DMSO: d6): 8.1(1H,
–
s, ArH, br), 7.9 (1H, s, ArH, br), 7.5 (2H, d, ArH, j = 7), 7.30 (2H, t,
ArH, j = 7), 7.1 (1H, t, ArH, j = 7), 5.67 (1H, s, NH, br), 2.08 (3H, s,
CH3).
2.2.2.4. 4-(4-(p-tolylamino)−6–chloro-1,3,5-triazin-2-yloxy)−2-
methoxybenzaldehyde (5d). C18 H15ClN4O3; Yield: 87%, White
2.2.5.2. 4–chloro-N-ethyl-6-(4-phenyl-1H-pyrazol-1-yl)−1,3,5-triazin-
2-amine(10b). C14 H13ClN6; Yield: 60%, White Solid, m.p. 125–
Solid, m.p. 140–143 °C, IR (KBr, Cm−1), 3363 (N H, st), 3067,
–
126 °C, IR (KBr, Cm−1), 3278 (N H, st), 3103 (C H, aromatic,
–
–
–
–
–
(C H, aromatic, st), 2936 (C H, aliphatic, st) 2816 (C H, aldehyde,
–
st), 2948 (C H, aliphatic, st), 1650 (C = N), 1400–1600 (C = C
st), 1717 (C = O, aldehyde, st), 1579 (C = N), 1371–1600 (C = C
aromatic), 730 (C Cl, st). 1H NMR: (400 MHz; DMSO: d6): 8.3 (1H,
aromatic), 1252 (C O, st), 770 (C Cl, st). 1H NMR: (400 MHz;
DMSO: d6): 11.12 (1H, s, CHO), 7.89(1H, d, ArH, j = 8), 7.73 (1H,
s, ArH), δ 7.10–7.30 (5H, m, ArH), 5.7 (1H, NH, br), 3.70 (3H, s,
OCH3), 1.25 (3H, s, CH3).
–
–
–
s, ArH, br), 7.9 (1H, s, ArH, br), 7.8 (2H, d, ArH, j = 6.9), 7.4 (2H,
t, ArH, j = 6.9), 7.2 (1H, t, ArH, j = 6.9), 4.83 (1H, s, NH, br), 2.90
(2H, q, CH2, j = 3), 1.20 (3H, t, CH3, j = 3).
2.2.5.3. N2,N4-dimethyl-6-(4-phenyl-1H-pyrazol-1-yl)−1,3,5-triazine-
2,4-diamine (10c). C14 H15N7; Yield: 60%, White Solid, m.p. 115–
2.2.3. General procedure for the synthesis of mono or diamino
substituted triazine compounds 8a-8d
116 °C, IR (KBr, Cm−1), 3278 (N H, st), 3086 (C H, aromatic,
–
–
The mono amino (ethyl or methyl) substituted derivatives (8a
and 8b) were synthesized using the protocol reported for com-
pound 3a-3d [26]. In diamino substituted triazine compounds, two
chlorine groups of the cyanuric chloride were replaced by amino
(ethyl or methyl) groups. For this purpose, trichloro-1,3,5-triazine
(5.0 g, 27 mmol) was dissolved in acetone (35 mL) and its temper-
ature keeps at 0–5 °C by putting it in an ice-water bath. Then a
solution of methylamine or ethylamine (54 mmol) was added in a
dropwise manner and the solution was stirred for 30 min at room
temperature. To this mixture, NaHCO3 (54 mL, 2 N 108 mmol) was
added and stirred for 6 h at 40 °C. Finally, the product as a white
powder was filtered, washed with water, and dried under vacuum
[26].
–
st), 2948 (C H, aliphatic, st), 1650 (C = N), 1400–1600 (C = C
aromatic). 1H NMR: (400 MHz; DMSO: d6): 8.4 (1H, s, ArH, br), 8.1
(1H, s, ArH, br), 7.54 (2H, d, ArH, j = 8), 7.22 (2H, t, ArH, j = 8),
7.15 (1H, t, ArH, j = 8), 5.20 (2H, s, 2NH, br), 2.50 (6H, s, 2CH3).
2.2.5.4. N2,N4–diethyl-6-(4-phenyl-1H-pyrazol-1-yl)−1,3,5-triazine-
2,4-diamine (10d). C16 H19 N7; Yield: 56%, White Solid, m.p. 131–
133 °C, IR (KBr, Cm−1), 3266 (N H, st), 3096 (C H, aromatic,
–
–
–
st), 2983 (C H, aliphatic, st), 1618 (C = N), 1399–1550 (C = C
aromatic). 1H NMR: (400 MHz; DMSO: d6): 8.1 (1H, s, ArH, br),
7.9(1H, s, ArH, br), 7.68(2H, d, ArH, j = 8), 7.4 (2H, t, ArH, j = 8),
7.2 (1H, t, ArH, j = 8), 4.53 (2H, s, 2NH, br), 3.00 (4H, q, 2CH2,
j = 3), 1.10 (6H, t, 2CH3, j = 3).
2.2.4. Synthesis of 4-Phenyl-1H-pyrazole (9)
4-Phenyl-1H-pyrazole (9) was synthesized by the earlier re-
2.2.5.5. 2,4-dichloro-6-(4-phenyl-1H-pyrazol-1-yl)−1,3,5-triazine
(10e). C12H7Cl2N5; Yield: 60%, White Solid, m.p. 110–112 °C, IR
ported procedure [27]. To
a dry and cooled DMF (25 mL),
POCl3 (10 mL) was added dropwise. Then phenylacetic acid (5 g,
37 mmol) was added and the solution was stirred at 85 °C for 3 h.
After cooling to room temperature, the mixture was poured onto
50 mL of ice and kept for 0.5 h. Then a saturated solution of NaBF4
(KBr, Cm−1), 3093 (C H, aromatic, st), 1560 (C = N), 1400–1600
–
(C = C aromatic), 742 (C Cl). 1H NMR: (400 MHz; DMSO: d6):
–
8.4(1H, s, ArH), 8.1 (1H, s, ArH), 7.70 (2H, d, ArH, j = 7.9), 7.4 (2H,
t, ArH, j = 7.9), 7.3 (1H, t, ArH, j = 7.9).
4

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
**
***
***
***
***
Fig. 1. The anti-inflammatory activity of vanillin-triazine derivatives in the carrageenan-induced paw edema test in the rat (n = 6). Vehicle (0.5% Tween 80 in saline) and
two doses of each compound (100 and 200 mg/kg as shown in parenthesis) were administered 30 min before to sub plantar injection of carrageenan, and the volume of the
paw (mL) was measured immediately before carrageenan injection and 4 h after injection. Indomethacin (10 mg/kg, i.p.) was used as the reference drug. Data are mean
SEM of six animals in each group. ∗P <0.05 and ∗∗∗P < 0.001 and show significant difference from the control group (ANOVA with Scheffe post hoc test).
2.3. Biological activity
+2 manually. This structure was protonated using the AutoDock
Tools program and was used for docking. The synthesized com-
pounds 5a-5d and 10a-10e were drawn with the hyperchem soft-
ware and optimized by using the semi-empirical PM3 method.
Then the most stable conformation was utilized in docking calcula-
tion. The AutoDock Tools program was used to generate the dock-
ing input files. For docking of the synthesized compounds into the
COX-1 and COX-2 structures, Auto Dock 1.5.6 software was used
[31]. The size of the binding pocket was detected 50 × 50 × 50
points for both enzymes. According to the position of celecoxib in
these structures, for the COX-2 enzyme, x (31.144), y (−22.960) and
z (−16.605) dimensions were built, while x (−32.344), y (43.754),
and z (−4.433) dimensions were used in COX-1 enzyme docking
studies. The grid and docking parameter was set according to de-
fault parameters and finally, the obtained results files were ana-
lyzed using Accelrys Discovery Studio Visualizer 3.0 program and
PyMOL Molecular Graphics System. By re-docking of celecoxib as
a co-crystallized inhibitor back into respective enzymes COX-2 and
2.3.1. Anti-inflammatory activity
The anti-inflammatory activity of the newly synthesized com-
pounds was evaluated using the carrageenan-induced paw edema
model in rats [28]. Experiments were performed on male Wistar
rats, weighing 200–220 g. All animals were maintained under stan-
dard laboratory conditions in the animal house of the School of
Pharmacy, Isfahan University of Medical Sciences (Isfahan, Iran).
These animals were euthanized immediately after each experi-
ment. All experiments were carried out by local guidelines for the
care of laboratory animals of Isfahan University of Medical Sciences
(Isfahan, Iran). Animals are weighed, randomized into 6 groups,
and kept for 1 week to acclimatize to the laboratory conditions and
to minimize the stress level. The synthesized compounds were ad-
ministered intraperitoneally (i.p.) at doses of 100 and 200 mg/kg to
rats. The control group received only vehicles. The reference group
or positive control received indomethacin (10 mg/kg). Thirty min-
utes after administration, the rats received a sub-plantar injection
of 100 μL of a 1% (w/v) suspension of carrageenan lambda in the
right hind paw [29]. The volume of the paw was measured by a
mercury plethysmometer (Ugo Basil, Italy) immediately before and
4 h after carrageenan injection. The data were expressed as mean
SEM of the volume difference (mL) of carrageenan-treated and
control paw. The data obtained in the experimental groups were
analyzed by one-way analysis of variance (ANOVA) followed by a
Scheffe post hoc test. P <0.05was considered significant. The result
of the anti-inflammatory activity of compounds is shown in Figs. 1
and 2.
˚
COX-1 with root mean square deviation (RMSD) of below 1 A val-
ues, the molecular docking protocol was validated. The free ener-
gies of binding (ꢀGb) and inhibition constants (Ki) and H-bond in-
teraction obtained from the docking studies of the compounds as
well as celecoxib as reference ligand with COX-1 and COX-2 by us-
ing Autodock4 is presented in Table 1.
3. Results and discussion
3.1. Preparation of novel vanillin-triazine and phenylpyrazole-triazine
derivatives
2.4. Molecular docking
Both vanillin-triazine and phenylpyrazole-triazine derivatives
were prepared from the multistep condensation reaction of
cyanuric chloride (1) and different nucleophiles as shown in
Scheme 2. Because of the different reactivity of three chloro groups
on triazines, the reactions were performed at different tempera-
tures. At low temperature (below 5 °C) only the first chloro of cya-
nuric chloride can be substituted, while at room temperature two
of them can participate in substitution reactions, and under high
temperatures, all three chloro groups can be replaced with nucle-
ophiles [32]
Molecular docking studies were performed with AutoDock 4.2.
The methodology used for molecular docking protocol was sim-
ilar to our previous reports [30]. The structures of COX-2 (PDB
˚
code: 3LN1, resolution: 2.4 A) and COX-1 (PDB code: 3KK6, resolu-
˚
tion: 2.75 A) were obtained from the protein databank. Celecoxib
as a co-crystallized inhibitor and water molecules were removed
together with β-octylglucoside and N-acetyl-d-glucosamine in the
structure; at the same time. The charge of the Fe atom was set to
5

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
*
**
*
**
***
***
***
***
***
***
Fig. 2. The anti-inflammatory activity of phenylpyrazole-triazine derivatives in the carrageenan-induced paw edema test in the rat (n = 6). Vehicle (0.5% Tween 80 in saline)
and two different doses of each compound (100 and 200 mg /kg as shown in parenthesis) were administered 30 min before to subplantar injection of carrageenan and the
volume of the paw (mL) was measured immediately before carrageenan injection and 4 h afterward. Indomethacin (10 mg/kg, i.p.) was used as the reference drug. Data are
mean
SEM of six animals in each group. ∗P <0.05 and ∗∗∗P < 0.001 show significant differences from the control group (ANOVA with the Scheffe post hoc test).
Table 1
Free binding energy (kcal/mol), interactions as well as inhibition constants (Ki) of synthesized compounds calculated by AutoDock.
COX2
Binding energy1
COX1
Binding energy1
compound
Inhibition constants
H-bond
Hydrophobic interaction
Inhibition constants
H-bond
2
2
5a
−8.69
−8.78
−9.23
−9.12
−7.14
−7.4
423.9
Arg499
Arg 499
PHe504
Arg499
Tyr355, Ser530, Leu352
−8.34
−7.27
−8.38
−7.74
−6.6
768.30
–
2
5b
367.85
Ser530, Tyr348, Val349
4.73(μm)
–
2
2
5c
172.25
Leu352, Ser530, Val349
423.08
–
2
3
5d
205.36
His90, Ser530, Leu352, tyr348
Ser530, Phe518, Leu531
2.11
–
10a
10b
10c
10d
10e
5.873
3.773
18.683
54.513
41.223
28.183
21.103
–
Leu352, Ile523, Tyr355, Trp387
Leu352, Tyr355, Ile523, Trp387
Phe518, Ile523, Tyr355, Ala527
Ser530, His90, Ser353, Tyr355,
−5.89
−5.98
−6.21
−6.41
Tyr371
3
−7.27
−7.41
−8.41
4.66
Ser530
Ser530
Tyr371
3
3.69
–
–
3
668.67
1
: (kcal/mol),.
2
: nanomolar,.
3
: micromolar.
tively. The absorptions at 2259 and 770–749cm−1 are related to
In vanillin-triazine series, substituted 4,6-dichloro-1,3,5-triazin-
2-ylphenylamine derivatives (5a-d) were synthesized by nucle-
ophilic attack of different aromatic amines(2a-2d) to cyanuric chlo-
ride (1) at 0–5 °C. In the next step disubstituted s-triazine com-
pounds (5a-5d) were obtained by the reaction of vanillin (4)
with 3a-3d derivatives. In phenylpyrazole-triazine compounds, one
and/or two chlorine of cyanuric chloride (1) was replaced by
methylamine (6) and/ or ethylamine (7) to produce (8a-8f) and the
next chloro was replaced with 4-Phenyl-1H-pyrazole (9) which was
prepared through Vilsmeier reaction, using a protocol reported in
earlier work to obtain final products (10a-10e). For the preparation
of tri-substituted triazines, the reactions were performed at 60 °C.
≡
–
N and C Cl groups, respectively. FT-IR spectrum of compound
C
5c is shown as an example in Fig. 3. The 1H NMR spectral data of
the final vanillin-triazine products recorded in (D6) DMSO along
with its possible assignments was reported in the experimental
part. All the aromatic and aliphatic H-atoms were found in their
expected regions. The absorption of aromatic H-atoms of vanillin
and phenyl rings appeared in the range of 8.1–7.2 ppm. The NH
of the phenylamine ring exhibited a broad singlet at 6.7–5.7 ppm.
The H-atoms of aldehyde and methoxy groups on the vanillin ring,
present in derivatives, were found in the region at 11.20–11.12 and
3.72–3.69 ppm, respectively. 1HNMR spectrum of compound 5c is
shown as an example in Fig. 4. It should be mentioned that the H-
atoms of methyl substitution on the phenyl rings in 5d, was found
in the region at 1.25 ppm.
3.2. Characterizations of synthesized compounds
FT-IR spectrum of phenylpyrazole-triazine compounds 10a-10e
also confirmed the structure of synthesized compounds. The main
IR vibrational bands of the compounds appeared around 1650–
1552 and 1600–1400 confirm the presence of C = N and C = C
groups, respectively. In derivatives containing the methylamine or
ethylamine group, absorptions at 2983–2921 and 3278–3221 cm−1
Novel vanillin-triazine and phenylpyrazole-triazine derivatives
were characterized by spectral techniques such as 1H NMR and FT-
IR techniques (the results are presented in the experimental part).
In the vanillin-triazine series, the FT-IR spectrum of compound
5a-5d showed absorptions at 3362–3270 and 1671–1557 cm−1
which indicate the presence of NH and C = N groups, respec-
tively. The vibrations in the range of 1717–1711cm−1 and also
2824–2742 cm−1, showed stretch vibration of the aldehyde group
of vanillin. The bands at 3132–3062 and 2936–2925 cm−1 also
–
indicated the presence of the aliphatic C H and NH in the struc-
tures. FT-IR spectrum of compound 10b is shown as an example in
Fig. 3. The chemical structure of the synthesized phenylpyrazole-
triazine compounds was further confirmed by the NMR technique.
–
confirm stretch vibration of aromatic and aliphatic C H, respec-
6

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
Fig. 3. FT-IR spectrum of compounds (a) 5c and (b) 10b.
In this series, the absorption of aromatic H-atoms of derivatives
appeared in the range of 8.4–7.1 ppm. In the 1H NMR spectrum of
compounds 10a-10f, the NH group of methyl and or ethylamine
was presented at 5.67–4.53 ppm. Methyl and methylene groups
related to methyl and ethylamine also were observed at 2.5–1.1
and 3.0–2.9 ppm, respectively. 1HNMR spectrum of compound 10b
is shown as an example in Fig. 4. To summarize, the NMR spec-
tra showed characteristic peaks for the synthesized hybrid com-
pounds.
activity (reduced carrageenan-induced paw inflammation by 93%)
at doses of 100 and 200 mg/kg significantly (P < 0.001). In
vanillin-triazine derivatives, compounds 5a and 5b didn’t exert
good anti-inflammatory properties in comparison to Indomethacin
as illustrated in Fig. 1. In this series, compound 5c at doses of
200 mg/kg significantly (P < 0.001) showed comparable anti-
inflammatory effects to indomethacin by a significant reduction
in paw size (81%) after carrageenan injection. This compound at
a dose of 100 mg/kg inhibited carrageenan-induced paw edema
by 54% (Fig. 1). In this series of compounds, 5d doses of 100
and 200 mg/kg resulted in significantly (P < 0.001) inhibition
of edema of 69% and 74% respectively. Phenylpyrazole-triazine
derivatives (10a-10e) showed better anti-inflammatory effects than
the vanillin-triazine derivatives 5a-5d Compound 10a at a dose
of 200 mg/kg significantly (P < 0.001) showed comparable anti-
inflammatory effects by a significant reduction in paw size (89.6%)
after carrageenan injection. Compound 10b at doses of 100 and
200 mg/kg significantly (P < 0.001) showed a greater inhibitory
effect (93%) than indomethacin and it was the best compound in
both series. Phenylpyrazole-triazine derivatives compound 10e at
doses of 200 mg/kg significantly (P < 0.001) showed comparable
anti-inflammatory effects by a significant reduction in paw size
(85%) after carrageenan injection.
3.3. Anti-inflammatory activity
A well-known model of acute inflammation is Carrageenan-
induced paw edema which consists of a biphasic inflammatory
response produced by several inflammatory mediators [33]. Af-
ter the injection of carrageenan into the rat paw, several medi-
ators such as histamine, serotonin, and bradykinin are sequen-
tially released in the initial phase (0–1 h), and then the produc-
tion of prostaglandins (PGs) are increased through the activation
of cyclooxygenase-2 (COX-2) and the release of nitric oxide (NO)
in the second phase (1–6 h) which are considered as inflammatory
factors and play important roles in the tissue damage by inflamma-
tion [33,34]. The earlier work showed that derivatives of s-triazine
exerted anti-inflammatory effects. About 35.17% of evaluated tri-
azines revealed promising which is related to the inhibition of the
p38 MAPK pathway or in the production of several inflammatory
mediators [18]
Data from Carrageenan-induced paw edema models suggest
that the designed triazine derivatives showed a very good anti-
inflammatory effect compared to the similar previously studied
compounds, reported in the Table 2. The anti-inflammatory effect
of the compounds may be due to a decrease in the production of
PGs, NO, bradykinin, or other inflammatory mediators. However,
more investigations are needed to find out their exact mechanism
of action.
In this investigation Indomethacin as
a
standard anti-
inflammatory drug inhibited carrageenan-induced paw edema
by 90% [Figs. 1 and 2]. Compound 10e as the best derivatives
in the studied compounds showed significant anti-inflammatory
7

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
Fig. 4. 1HNMR spectrum of compounds (a) 5c and (b) 10b in DMSO.
3.4. Discussion of docking
derivatives, and balanced its placing with the key amino acid,
Arg499, by forming proper H-bonds. Meanwhile, some H-bonds
are stabilized by the binding of the Phe504 amino acids with
the carbonyl oxygen of the vanillin in aminobenzonitryl derivative.
The phenyl ring of aniline derivatives and vanillin formed proper
hydrophobic interactions with His90, Tyr355, Ser530, Tyr385,
and Leu352.
One of the computationally inexpensive methods for predict-
ing if and how a ligand will bind to a protein binding pocket,
followed by an estimation of how strong is the ligand-binding
affinity, is molecular docking [40-42]. To explain the COX-1/COX-
2 selectivity of newly synthesized compounds, a docking study
was used to estimate the possible inhibitory activities and mecha-
nism of binding to COX enzymes. In general, the binding energy of
both synthetic compounds with cyclooxygenase II was higher than
the cyclooxygenase I enzyme. The result of docking of vanillin-
triazine derivatives showed a relatively better selectivity to COX-
II active site which can be attributed to having more functional
groups on this group of compounds. In vanillin-triazine deriva-
tives by holding constant the aniline core, substitution of bromine,
methyl, and nitrile groups on phenyl rings may increase bind-
ing energy through hydrophobic and H-bonding interactions. The
aldehyde groups on the vanillin ring played a conserved role in
stabilizing the docking of aniline, bromoaniline, and methylamine
Phenylpyrazole-triazine derivatives also showed better selectiv-
ity to COX-II active site. In this series, when two methylamine
or two ethylamine groups are placed on a tri-azine ring, the
compound can form a hydrogen bond with Ser530 amino acid.
The triazine and phenylpyrazole ring in his group also formed
proper hydrophobic interactions with His90, Tyr355, Typr387, and
Leu352.
Molecular docking studies have shown that compounds 4-
aminobenzonitryl derivative from vanillin-triazine series, 5c (ꢀG
binding= −9.23) and aminoethyl triazine- phenylpyrazole, 10c (ꢀG
binding= −7.41) are the best compounds in each category, respec-
tively (Fig. 5).
8

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
Table 2
Anti-inflammatory effect of similar previously studied compounds.
Triazine based
compound
Anti-inflammatory
test
Route of
% Inhibition of
Edema.
Degree of paw
administration
Std. drug
thickness
Ref.
7-methyl-4-
phenylpyrazolo[1,5-
α][1,3,5]triazine-
2(8H)-
Carrageenan
induced rat paw
edema
oral
Phenylbutazone
Std.=60
–
[35]
Comp= 54.28
thione
N-(2, 6-di
Carrageenan
induced rat paw
edema
oral
Diclofenac
Std.=63.00
–
[36]
(piperazin-1-yl)
pyrimidin-4-yl)−1-
(4,
Comp=68.00
8-dimethoxy-3-
methylbenzo
[1,2-b: 5,4-b‘]
difuran-2-yl)−1-
(piperazin-1-yl)
methanimine
N-arylidene-N’- [5-
(4-isobutylphenyl)-
[1,2,4]-triazin-3-yl]
hydrazines
Carrageenan
induced rat paw
edema
oral
oral
Ibuprofen
Std.= 50.51
–
–
[37]
[38]
Comp=58.37
2- (5,6-diphenyl-3-
thioxo-1,2,4-
triazine-2-
Carrageenan
induced rat paw
edema
Indomethacin
Std.= 79.28
Comp=58.24
yl)−3,4,5-
trimethoxy-
phenilic
acid
vanilline
Carrageenan
induced rat paw
edema
i.p.
ip
diclofenac
–
–
Std.=0.4
[19]
[39]
Comp=0.4
(2,3-Dihydro-
benzofuran-5-yl)-
acetic acid
Carrageenan
induced rat paw
edema
Diclofenac sodium
Std.=1.16
Comp=1.00
ꢀ
[4-(4 -fluoro-
biphenyl-4-
ylmethoxy)−3-
methoxybenzylidene]-
hydrazide
ip: Intraperitoneal injection.
–: not reported.
Fig. 5. The binding mode of compounds 5c (A) and 10c (B) in the active site of COX-II was obtained from autodock4.
4. Conclusion
methods (FT-IR, and NMR spectral data). The anti-inflammatory ac-
tivity of these derivatives was evaluated using the carrageenan-
induced paw edema model in male Wistar rats (200–220 g) ad-
ministered intraperitoneally at doses of 100 and 200 mg/kg. Com-
pounds 5c, 5d, 10a, 10e at doses of 200 mg/kg significantly (P
Two series of triazine-based derivatives were synthesized and
evaluated for in vivo anti-inflammatory activities. The structure of
nine synthesized compounds (5a-5d and 10a-10e) consisting of tri-
azine, vanillin, and phenylpyrazole was confirmed by spectroscopic
<
0.001) showed anti-inflammatory effects comparable to the
9

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
standard drug indomethacin (90%) by a significant reduction in
paw size (74–89.6%) after carrageenan injection. Compound 10b
at doses of 100 and 200 mg/kg significantly (P < 0.001) showed
a greater anti-inflammatory activity (reduced carrageenan-induced
paw inflammation by 93%) than indomethacin and it was the best
compound in both series. From the docking studies, compounds 5c
and 10c are the most active analogs, with the lowest binding ener-
gies. These two compounds exhibited inconsequential recognition
at the binding pocket of COX-II. Further studies are necessary to
understand the underlying and implicated mechanisms of the ob-
served pharmacologic effects.
[14] M.M.M. El-Miligy, A.A. Hazzaa, H. El-Messmary, R.A. Nassra, S.A.M. El-Hawash,
New hybrid molecules combining benzothiophene or benzofuran with rhoda-
nine as dual COX-1/2 and 5-LOX inhibitors: synthesis, biological evaluation and
docking study, Bioorg. Chem. 72 (2017) 102–115.
[15] L.W. Mohamed, O.M. El-Badry, A. El-Ansary, A. Ismael, Design & synthesis of
novel oxazolone
& triazinone derivatives and their biological evaluation as
COX-2 inhibitors, Bioorg. Chem. 72 (2017) 308–314.
[16] A.G. Banerjee, N. Das, S.A. Shengule, R.S. Srivastava, S.K. Shrivastava,
Synthesis, characterization, evaluation and molecular dynamics studies
of 5, 6-diphenyle1,2,4etriazine3(2H)-one derivatives bearing 5-substituted
1,3,4-oxadiazole as potential anti-inflammatory and analgesic agents, Euro. J.
Med. Chem. 101 (2015) 81–95.
[17] H.A.H. Elshemy, E.K.A. Abdelall, A.A. Azouz, A. Moawad, W.A.M. Ali,
N.M. Safwat, Synthesis, anti-inflammatory, cyclooxygenases inhibitions assays
and histopathological study of poly-substituted 1,3,5-triazines: confirmation
of regiospecific pyrazole cyclization by HMBC, Eur. J. Med. Chem 127 (2017)
10–21.
Declaration of Competing Interest
[18] L. Marin-Ocampo, L.A. Veloza, R. Abonia, J.C. Sepulveda-Arias, Anti-inflamma-
tory activity of triazine derivatives: a systematic review, Euro. J. Med. Chem.
162 (15) (2019) 435–447.
None.
[19] J. Niazi, N. Kaur, R.K. Sachdeva, Y. Bansal, V. Gupta, Anti-inflammatory and
antinociceptive activity of vanillin, Drug Develop. Therap. 5 (2) (2014) 143–147.
[20] Yu.A. Boiko, M.V. Nesterkina, A.A. Shandra, I.A. Kravchenko, Analgesic and An-
ti-Inflammatory Activity of Vanillin Derivatives, Pharm. Chem. J. 53 (2019)
650–654.
CRediT authorship contribution statement
Parvin Asadi: Software, Visualization, Writing – original draft,
[21] S. Celik, F. Ozkok, A.E. Ozel, E. Cakir, S. Akyuz, F.T.-.I.R. Synthesis, N.M.R. Char-
acterization, Antibacterial and Antioxidant Activities, and DNA Docking Anal-
ysis of a New Vanillin-Derived imine Compound, J. Mole. Struct. 1236 (15)
(2021) 130288.
Conceptualization, Validation, Formal analysis, Resources, Writing
–
review & editing. Mohsen Alvani: Writing – original draft,
Formal analysis, Investigation. Valiollah Hajhashemi: Conceptual-
ization, Validation, Formal analysis, Writing – review & editing.
Mahboubeh Rostami: Conceptualization, Validation, Formal anal-
ysis. Ghadamali Khodarahmi: Supervision, Project administration,
Conceptualization, Validation, Resources, Writing – review & edit-
ing.
[22] C. Barja-Fidalgo, I.M. Fierro, A.C.B. Lima, E.T.D. Silva, C.D.A. Camara, E.J. Bar-
reiro, Anti-inflammatory Profile of N-Phenylpyrazole Arylhydrazone Derivatives
in Rats, J. Pharm. Pharmacol 51 (1999) 703–707.
[23] K. Ando, M.L. Minich, J. Li, S.M. Sakya, K.M. Lundy, H. Cheng, B.S. Bronk,
K. Kawamura, T., Kato, Heteroaryl phenyl pyrazole compounds as anti-
inflammatory/analgesic agents, US6649636B1 United States, (2003)
[24] E. Banoglu, C. Akoglu, S. Unlu, E. Kupeli, E. Yesilada, Amide derivatives
of [6-(5-Methyl-3-phenylpyrazole-1-yl)-3(2H)-pyridazinone-2-yl]acetic acids as
potential analgesic and anti-inflammatory compounds, Arch Pharma 337 (1)
(2004) 7–14.
References
[25] H.R. Bhat, U.P. Singh, P. Gahtori, S.K. Ghosh, K. Gogoi, A. Prakash, R.K. Singh,
Antimalarial activity and docking studies of novel bi-functional hybrids
derived from 4-aminoquinoline and 1,3,5-triazine against wild and mu-
[1] S.A. Khan, S.M. Imam, A. Ahmad, S.H. Basha, A. Husain, Synthesis, molecular
docking with COX 1 & II enzyme, ADMET screening and in vivo anti-inflam-
matory activity of oxadiazole, thiadiazole and triazole analogues of felbinac, J.
Saudi. Chem. Soc. 22 (2018) 469–484.
tant malaria parasites as pf-DHFR inhibitor, RSC Adv
3 (9) (2013)
2942–2952.
[26] X.Z. Zheng, J.L. Zhou, J. Ye, P.P. Guo, C.S. Lin, Cardioprotective effect of novel
sulphonamides-1,3,5-triazine conjugates against ischaemic–reperfusion injury
via selective inhibition of MMP-9, Chem. Biol. Drug Design. 88 (5) (2016)
756–765.
[2] A.A. Fadda, K.M. Elatta, Design and synthesis of some enaminonitrile deriva-
tives of antipyrine as potential novel anti-inflammatory and analgesic agents,
J. Biosci. Med. 3 (2015) 114–123.
[3] H. Yoshimura, S. Sekine, H. Adachi, Y. Uematsu, A. Mitani, N. Futaki, N. Shimizu,
High levels of human recombinant cyclooxygenase-1 expression in mammalian
cells using a novel gene amplification method, Protein Expr. Purif. 80 (2011)
41–46.
[27] J. Olguin, S. Brooker, Synthesis of 3- and 5-formyl-4-phenyl-1H-pyrazoles:
promising head units for the generation of asymmetric imine ligands
and mixed metal polynuclear complexes, New J. Chem. 35 (2011)
1242–1253.
[4] C.S. Manohar, A. Manikandan, P. Sridhar, A. Sivakumar, B.S. Kumar, S.R. Reddy,
Drug repurposing of novel quinoline acetohydrazide derivatives as potent
COX-2 inhibitors and anti-cancer agents, J. Mol. Structure. 1154 (2018)
437–444.
[28] K.H. Kim, H.W. Im, M.B. Karmacharya, S. Kim, B.H. Min, S.R. Park, B.H. Choi,
Low-intensity ultrasound attenuates paw edema formation and decreases vas-
cular permeability induced by carrageenan injection in rats, J. Inflam. 17 (1)
(2020) 1–8.
[29] L. Rafiee, V. Hajhashemi, S.H. Javanmard, Maprotiline inhibits COX2 and iNOS
gene expression in lipopolysaccharide-stimulated U937 macrophages and car-
rageenan-induced paw edema in rats, Central-Euro, J. immune. 44 (1) (2019)
15.
[5] C.S. Li, C. Brideau, C.C. Chan, C. Savoie, D. Claveau, S. Charleson, R. Gordon,
G. Greig, J.Y. Gauthier, C.K. Lau, D. Riendeau, M. Thérien, E. Wong, P. Pra-
sit, Pyridazinones as selective cyclooxygenase-2 inhibitors, Bioorg. Med. Chem.
Lett. 24 (2003) 597–600.
[6] I.G. Rantish, K. Javed, K. Ahmed, S. Bano, M.S. Alam, K.K. Pillai, S. Singh,
V. Bagchi, Synthesis and anti-inflammatory activity of some new 1,3,5- trisub-
stituted pyrazolines bearing benzene sulfonamide, Bioorg. Med. Chem. Lett. 19
(2009) 255–258.
[7] A. Bertolini, A. Ottani, M. Sandrini, Selective COX-2 inhibitors and dual act-
ing anti-inflammatory drugs: critical remarks, Curr. Med. Chem. 9 (10) (2002)
1033–1043.
[8] K.M. Qiu, R. Yan, M. Xing, et al., Synthesis, biological evaluation and molecu-
lar modeling of dihydro-pyrazolyl-thiazolinone derivatives as potential COX-2
inhibitors, Bioorg. Med. Chem. 20 (22) (2012) 6648–6654.
[30] M. Dinari, F. Gharahi, P. Asadi, Synthesis, spectroscopic characterization, an-
timicrobial evaluation and molecular docking study of novel triazine-quinazoli-
none based hybrids, J. Mole. Struct. 1156 (2018) 43–50.
[31] R. Huey, G.M. Morris, in: Using AutoDock 4 with AutoDocktools: a Tutorial, The
Scripps Research Institute, USA, 2008, pp. 54–56.
[32] M. Dinari, M.M. Momeni, M. Afshari, Fabrication and characterization of hy-
brid films based on polyaniline and graphitic carbon nitride nanosheet, J. Appl.
Polym. Chem. 133 (2016) 44059.
[33] V. Hajhashemi, H. Mojiri-Froshani, L. Saghaei, A. Fassihi, A study on the an-
ti-inflammatory effects of new derivatives of 3-hydroxy pyridine-4-one, Adv.
Biomed. Res. 3 (2014) 134.
[9] D. Dogne, C.T. Supuran, D. Pratico, Adverse cardiovascular effects of the coxibs,
J. Med. Chem. 48 (2005) 2251–2257.
[34] M. Zabihi, V. Hajhashemi, M. Minaiyan, A. Talebi, Evaluation of the central
and peripheral effects of doxepin on carrageenan-induced inflammatory paw
edema in rat, RPS 12 (4) (2017) 337.
[10] G. Mariappan, B.P. Saha, L. Sutharson, et al., The diverse pharmacological im-
portance of pyrazolone derivatives, J. Pharm. Res. 3 (2010) 2856–2859.
[11] K. NAnkali, J. Rangaswamy, M. Shalavadi, N. Naik, G. Krishnamurthy, Synthesis
and Molecular Docking of novel 1,3-Thiazole Derived 1,2,3-Triazoles and In vivo
Biological Evaluation for their Anti anxiety and Anti inflammatory Activity, J.
Mol. Struct. 1236 (15) (2021) 130357.
[35] N.V.
Patel,
D.J.
Sen,
Synthesis
of
Substituted
7-Methyl-2,8-
Dihydropyrazolo-[1,5-α][1,3,5]-Triazine Derivatives for Anti-Inflammatory
and AntiMicrobial Screening, AJADD1 2 (2013) 157–173.
[36] A.A. Abu-Hashem, S.A. Al-Hussain, M.E.A. Zaki. Synthesis of Novel Benzodi-
furanyl; 1,3,5-Triazines;1,3,5-Oxadiazepines; and Thiazolopyrimidines Derived
from Visnaginone and Khellinone as Anti-Inflammatory and Analgesic Agents
Molecules. 25(2020), 220.
[12] N.S. Akhmadiev, E.S. Mescheryakova, V.R. Akhmetova, V.R. Khairullina,
L.M. Khalilov, A.G. Ibragimov, Synthesis, Crystal Structure and Docking Studies
as Potential Anti-Inflammatory Agents of Novel Antipyrine Sulfanyl Derivatives,
J. Mol. Struc. 1228 (15) (2021) 129734.
[37] A.A. Makhlouf, Y.A. Makladb, Synthesis and Analgesic-Antiinflammatory Activ-
ities of Some 1,2,4-Triazine Derivatives, Drug Res 54 (1) (2004) 42–49.
[38] P. Mullick, S.A. Khan, T. Begum, S. Verma, D. Kaushik, O. Alam, Synthesis
of 1,2,4-triazine derivatives as potential anti-anxiety and anti-inflammatory
agents, Acta Poloniae Pharm. - Drug Res. 66 (6) (2009) 379–385.
[13] A.G. Banerjee, N. Das, S.A. Shengule, P.A. Sharma, R.S. Srivastava, S.K. Shrivas-
tava, Design, synthesis, evaluation and molecular modelling studies of some
novel 5,6-diphenyl-1,2,4-triazin-3(2H)-ones bearing five-member heterocyclic
moieties as potential COX-2 inhibitors:
Bioorg. Chem. 69 (2016) 102–120.
a hybrid pharmacophore approach,
10

P. Asadi, M. Alvani, V. Hajhashemi et al.
Journal of Molecular Structure 1243 (2021) 130760
[39] L.V. Kumar Reddy, E. Kathale, Synthesis and Anti-Inflammatory Activity of Hy-
drazones Bearing Biphenyl Moiety and Vanillin Based Hybrids, Orient. J. Chem.
33 (2) (2017) 971–978.
[40] J.Q. Liu, X.F. Li, C.Y. Gu, J.C.S.d. Silva, A.L. Barros, S.A. Jr, B.H. Li, F. Ren, S.R. Bat-
ten, T.A. Soares, A combined experimental and computational study of novel
nanocage-based metal–organic frameworks for drug delivery, Dalton Trans. 44
(2015) 19370–19382.
[41] F. Li, B. Li, C. Wang, Y. Zeng, J. Liu, C.Y. Gu, P. Lu, L. Mei, Encapsulation of
pharmaceutical ingredient linker in metal–organic framework: combined ex-
perimental and theoretical insight into the drug delivery, RSC Adv. 6 (2016)
47959–47965.
[42] D.Y. Ma, Z. Li, J.X. Xiao, R. Deng, P.F. Lin, R.Q. Chen, Y.Q. Liang, H.F. Guo, B. Liu,
J.Q. Liu, Hydrostable and Nitryl/Methyl-Functionalized Metal–Organic Frame-
work for Drug Delivery and Highly Selective CO2 Adsorption, Inorg. Chem. 54
(14) (2015) 6719–6726.
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