A.R. Nesaragi et al.
Bioorganic & Medicinal Chemistry Letters 43 (2021) 128112
Fig. 3. COX-2 inhibition (%) of compounds 8i-t at different concentrations.
*STD- Mefenamic acid.
Fig. 2. RMSD of COX-2 enzyme and compound 8l.
compounds together, compound 8s and Mefenamic acid (standard) with
the enzyme (PDB ID: 5IKR; A-Chain), hydrophobic and hydrophilic
amino acids of the enzyme surrounding the compounds 8l and 8s are
provided in the supplementary information.
Table 3
COX-1/COX-2 inhibitory activity of the compounds 8i-t.
Compound
COX-1 (IC50 µg/mL)
COX-2 (IC50 µg/
Selectivity Index
We have executed the computation of Root Mean Square Deviation
(RMSD) which appraises resemblance flanked by two superimposed
atomic coordinates. The acquired plot of RMSD v/s simulation time of
COX-2 enzyme and compound 8l is embodied in Fig. 2, which suggested
that docking study was affirmative and our devised molecules man-
ifested significant affinity towards 5IKR _ligand.
mL)
8i
245.93 ± 1.36
104.24 ± 0.86
106.82 ± 1.16
435.26 ± 1.02
87.26 ± 3.52
506.80 ± 2.32
206.13 ± 2.62
576.87 ± 4.16
326.35 ± 1.53
283.58 ± 2.16
145.18 ± 1.08
245.32 ± 0.96
186.34 ± 1.21
19.95 ± 0.91
32.93 ± 1.35
24.59 ± 1.21
12.82 ± 0.98
23.74 ± 1.61
25.02 ± 1.14
54.42 ± 0.86
44.54 ± 2.10
17.70 ± 1.76
11.35 ± 1.52
13.67 ± 2.72
13.23 ± 1.04
25.21 ± 1.16
12.32
3.16
8j
8k
4.34
8l
33.95
3.67
8m
8n
20.25
3.78
8o
In vitro anti-inflammatory assay
8p
12.95
18.43
24.98
10.62
18.54
7.39
8q
8r
The synthesized molecules were evaluated for in vitro COX-1 and
COX-2 inhibition. The results obtained in the form of IC50 and are
tabulated in Table 3. All the compounds were active as COX-1 and COX-
2 inhibitors. It is evident from the obtained data that the synthesized
compounds are more selective as COX-2 inhibitors than COX-1 in cor-
relation with the docking studies, which revealed the title compounds
prompted better uniformity towards COX-2 than COX-1. The compounds
possessing halo substituent exhibited the most potent and discerning
inhibition of COX-2. The compounds 8i (methyl at C6 position of the
coumarin), 8l (benzo at C7,8 position of the coumarin), 8q (Cl at C5
position of benzoxazole and benzo at C5,6 position of the coumarin), 8r
(Cl at C5 position of benzoxazole and benzo at C7,8 position of the
coumarin), 8s (Cl at C5 position of benzoxazole and methyl at C7 posi-
tion of the coumarin) and 8t (Cl at C5 position of benzoxazole and
methyl at C5,7 position of the coumarin) emerged as most potent and
selective COX-2 inhibitors with IC50 values of 19.95, 12.82, 17.70,
11.35, 13.67 and 13.23 µg/mL respectively (Fig. 3). The selectivity
index of 8l, 8n and 8r was found to be 33.95, 20.25 and 24.98 respec-
tively, manifested their high selectivity against COX-2. Overall, this
study paves a way for the development of 2,5-substituted 1,3,4-oxa-
diazoles as potent and selective COX-2 inhibitors.
8s
8t
Mefenamic acid
COX-1 and COX-2 selectivity and their binding approaches. The com-
pounds were comprised in hydrogen bonding interactions with the hy-
droxyl group of SER530 which have been reported to play a vital part in
the inhibition of COX-2 enzyme.
The biological outcomes were well supported by the consequences of
the computational study where the docked compounds were instituted
to reinforce the examined inhibitory results. This suggests that the
compounds are originated to have elevated resemblance towards COX-2
enzyme than COX-1 thus exhibiting utmost selectivity towards COX-2.
The docking study divulged that compound 8l acts as good inhibitor
of cyclooxygenase 2 enzyme. As portrayed in Fig. 1, compound 8l
fabricated five hydrogen bonding interactions at the dynamic location of
the enzyme (PDB ID: 5IKR; A-Chain) out of which nitrogen atom present
on the 3rd position of oxadiazole ring frames two bonding interactions
with hydrogen atoms of ARG120 (-N—— H-ARG120, 2.09 Å; 1.75 Å),
the nitrogen atom present on the 4th position of oxadiazole ring makes
two interactions with hydrogen atoms of ARG120 (-N—— H-ARG120,
2.23 Å; 2.76 Å) and oxygen atom present on the 2nd position of chro-
mene ring forms an interaction with hydrogen atom of SER530 (-O——
H-SER530, 2.55 Å).
In vitro anti-oxidant assay
The reactive oxygen species play conclusive role in inflammatory
provisions. Oxidative stress is essential for gastrointestinal ulceration.
The compounds possessing antioxidant activity besides anti-
inflammatory activity may offer a feasible direction to more secure
anti-inflammatory agents.22
The consensus score acquired in the range 4.17–9.51 implied that
docking study was optimistic and the devised molecules manifested
substantial affinity towards 5IKR_ligand. The docked poses revealed that
the studied compounds have demonstrated the same kind of interaction
with amino acid residue (SER530) as that of 5IKR_ligand. Eventually, we
conclude that synthesized molecules preferentially bind to enzyme in
comparison to the 5IKR_ligand (Table 2) which substantiates the
observed preferential in vitro COX-2 inhibition. The interaction of all the
Antioxidants that evince DPPH radical scavenging activity is pro-
gressively accepting consideration as they have been reported to
encompass interesting anticancer, anti-ageing and anti-inflammatory
activities. Accordingly, compounds with antioxidant property could be
5