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

Article abstract of DOI:10.1016/j.molstruc.2021.130357

Heterocyclic rings such as thiazole and triazole are considered as privileged moieties, since they constitute several drugs for biological treatment. In this work, a novel series of 1, 3-thiazole linked 1,2,3-triazole derivatives were designed and synthesized according to convenient synthetic procedures. All the synthesized compounds are characterized by ¹H NMR, ¹³C NMR and LCMS techniques. The molecular docking was studied to illustrate the binding interactions of target molecules with GABA_A receptor. The synthesized compounds containing 1,3-thiazole and 1,2,3-triazole ring, the presence of these rings in each molecule may lead to have potential in vivo anti-anxiety and anti-inflammatory properties. The in vivo activity result revealed that some of the compounds possessed statistical significant therapeutic efficacy. Anti-anxiety screening on mice indicated that all the target compounds (5mg/kg) exhibited certain extent of an anxiolytic effect by increasing time spent on open arms and the percentage of open arm entries as compared to controlled group. More importantly, among the newly synthesized compounds (6h) and (6i) have strong anti-anxiety against mice. The non steroidal anti-inflammatory activity drugs (NSAIDs) are plays a very important role to prevent the growth of cyclooxygenase enzymes which are responsible for inflammation and pain. The results show that the following compounds 6e, 6g, 6h, and 6k are having maximum anti inflammatory activity against carrageenan induced acute inflammation in rats comparable to diclofenac as reference drug. Molecular docking simulations were employed to find out the important binding modes responsible for the anti anxiety activity, thus supporting their effective anti-anxiety efficacy.

Full text of DOI:10.1016/j.molstruc.2021.130357

Journal of Molecular Structure 1236 (2021) 130357
Contents lists available at ScienceDirect
Journal of Molecular Structure
journal homepage: www.elsevier.com/locate/molstr
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
a
a
b
a,∗
Kariyappa N Ankali , Javarappa Rangaswamy , Mallappa Shalavadi , Nagaraja Naik ,
c
Ganga naik Krishnamurthy
a
Department of Studies in Chemistry, University of Mysore, Manasagangotri, Mysuru, 570006, Karnataka, India
b
Department of Pharmacology, HSK College of Pharmacy, Bagalkot, 587101, Karnataka, India
c
Department of P.G. studies in Chemistry, Sahyadri Science College, A constituent college of Kuvempu University, Shivamogga, 577203, Karnataka, India
a r t i c l e i n f o
a b s t r a c t
Article history:
Heterocyclic rings such as thiazole and triazole are considered as privileged moieties, since they con-
stitute several drugs for biological treatment. In this work, a novel series of 1, 3-thiazole linked 1,2,3-
triazole derivatives were designed and synthesized according to convenient synthetic procedures. All the
synthesized compounds are characterized by 1H NMR, 13 C NMR and LCMS techniques. The molecular
docking was studied to illustrate the binding interactions of target molecules with GABAA receptor. The
synthesized compounds containing 1,3-thiazole and 1,2,3-triazole ring, the presence of these rings in each
molecule may lead to have potential in vivo anti-anxiety and anti-inflammatory properties. The in vivo
activity result revealed that some of the compounds possessed statistical significant therapeutic efficacy.
Anti-anxiety screening on mice indicated that all the target compounds (5mg/kg) exhibited certain extent
of an anxiolytic effect by increasing time spent on open arms and the percentage of open arm entries
as compared to controlled group. More importantly, among the newly synthesized compounds (6h) and
Received 5 September 2020
Revised 8 December 2020
Accepted 22 March 2021
Available online 26 March 2021
Keywords:
Thaizole
1,2,3-triazole
anti-inflammatory
anti-anxiety
molecular docking
(
6i) have strong anti-anxiety against mice. The non steroidal anti-inflammatory activity drugs (NSAIDs)
are plays a very important role to prevent the growth of cyclooxygenase enzymes which are responsible
for inflammation and pain. The results show that the following compounds 6e, 6g, 6h, and 6k are having
maximum anti inflammatory activity against carrageenan induced acute inflammation in rats comparable
to diclofenac as reference drug. Molecular docking simulations were employed to find out the impor-
tant binding modes responsible for the anti anxiety activity, thus supporting their effective anti-anxiety
efficacy.
©
2021 Elsevier B.V. All rights reserved.
1. Introduction
olytic compounds in which such activity has not yet been observed
4]. The pharmacotheraphy of anxiety disorders remains the most
[
Anxiety is the common neurological disorder characterized by
cognitive, emotional, and somatic behaviour changes. These symp-
toms may create an unpleasant feeling that is associated with
apprehension, fear which results in disturbance of daily routine
life [1]. The aetiology of these disorders is the dysfunction of
monoamine neurotransmitters in CNS, such as gamma-amino bu-
tyric acid (GABA), serotonin (5-HT), dopamine (DA) and nore-
pinephrine (NE) [2-3]. Therefore, to prevent the probable symp-
toms of neurological disorder, there is need to find novel anxi-
widespread and efficacious treatment, and the development of new
anti-anxiety drugs is trending active field. These psychiatric issues
are with an incidence of 18% and a life prevalence of 29% in the
population [5].
Inflammation is one of the common condition in which body
tissue are affected by heat, redness, swelling and pain. In order to
protect against injury, inflammation, the activation of macrophages
is an important part of the body immunity system. The activated
macrophages releases the many inflammatory cytokines, such as
interlukin-1ß (IL-1ß), interlukin-6 (IL-6) and tumor necrosis factor
(
TNF) are very essential in fighting against inflammation. An excess
release of these cytokines may causes acute and chronic diseases
6].
∗
Corresponding author at: Department of Studies in Chemistry, University of
[
Mysore, Manasagangotri, Mysuru, 570006, Karnataka, India.
E-mail address: drnaikchem@gmail.com (N. Naik).
https://doi.org/10.1016/j.molstruc.2021.130357
0
022-2860/© 2021 Elsevier B.V. All rights reserved.

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
Figure 1. The proposed structure of 1,3-thiazole derived 1,2,3-triazoles 6(e-p)
Heterocyclic organic chemistry is one of the most important
and well-studied branches of medicinal chemistry. An important
feature of heterocyclic bioactive compounds is their various con-
stituent heteroatoms, including nitrogen, sulphur, oxygen, and oth-
ers. The most promising heterocyclic compounds are azoles, which
are five-membered nitrogen heterocycles [7].
coated with silica gel 60F-254 with visualization under UV (254
nm) and SiO , 200-400 mesh (Merck) was used for column chro-
2
matography Hexane: Ethyl acetate (EA), 8:2 used as mobile phase.
1H NMR (400 MHz) and 13C NMR (100 MHz) was obtained AC
Bruker spectrometer in the appropriate (CDCl ) solvent. Melting
3
points were obtained on a reichert thermopan melting point ap-
paratus, equipped with a microscope and are uncorrected. Mass
spectra were obtained on an Electron Impact mass spectrometer
at 70 ev ionizing beam and using a direct insertion probe.
Triazole ring is the core structure and its derivatives are known
to exhibit various pharmacological properties such as, antimicro-
bial, antitubercular, anticancer, anticonvulsant, antiinflammatory,
analgesic and antiviral. Triazoles have also been incorporated in a
wide variety of therapeutically interesting drugs including H /H
2.2. Synthetic procedure
1
2
histamine receptor blockers, CNS stimulants, anti-anxiety agents
and sedatives. Many commercial antifungal drugs contain triazole
functional moiety. Triazoles are used in the control of variety of
fungal diseases in fruits, vegetables, legumes and grain crops, both
as pre- and postharvest applications [8].
2.2.1. General procedure for the synthesis of azidobenzene derivatives
3(e-p)
The substituted aniline derivatives 1(e-p)(1eq) was dissolved in
water and conc. HCl under vigorous stirring in an ice/water bath.
A freshly prepared cold sodium nitrite (1 eq) solution in water was
added drop wise to the reaction mixture by keeping the tempera-
ture between 0-5 °C. After completion of addition, stirred the reac-
tion for an additional 10 min, then freshly prepared sodium azide
(1 eq) solution in water was added slowly to the reaction mixture
via additional funnel while keeping the internal temperature of the
reaction mixture below 5 °C. After compete addition of the sodium
azide solution, the reaction mixture was stirred for about 30 min
at 0-5 °C, followed by stirring at room temperature for 2 hrs. The
reaction mixture was extracted with ethyl acetate (EtOAc) (25 ml)
three times. The combined organic layer was washed with water
(25 ml) and brine (25 ml), dried over anhydrous Na SO and con-
On the other hand, thiazole motif is a fascinating building block
in pharmaceutical chemistry for designing and synthesizing mul-
tiple bioactive agents [9-16]. Different studies investigated that
many thiazole analogues exhibited very potent antitumor activity
targeting specific pathways such as; suppressing microtubule poly-
merization as well as inhibition of various molecular targets such
as receptor tyrosine kinases (RTKs), non-receptor tyrosine kinases
(
nRTKs), phosphatidylinositol-3- kinases (PI3Ks), serine/threonine
kinases (STKs), Bcl-2 family, histone deacetylases (HDACs) and tu-
mor necrosis factor-α (TNF-α) [17-19]. In addition to this, The
substituted thaizole derivatives have been reported with various
biological activities such as anti-inflammatory, antipsychotropic,
antibacterial, antifungal, anticancer, antiallergic [20]. Compounds
containing substituted 1, 3-thiazole and 1, 2, 3-triazole heterocyclic
rings play a significant role in drug discovery and in both organic
and pharmaceutical chemistry. The rational design of new potent
biological agents of more selectivity towards various diseases takes
place through the molecular hybridization of two or more phar-
macophoric units recognized and derived from known bioactive
molecules to reach a final compound that would progress to clini-
cal trials and development. [17, 21].
2
4
centrated under reduced pressure to get crude azidobenze deriva-
tives 3(e-p), they were then purified by column chromatography
over silica gel (60-120 mesh) using hexane/EtOAc (9;1) as eluent.
The pure azidobenzene derivatives can stored at 2-5 °C in the re-
frigerator.
2.2.2. Synthesis of 1-phenylthiourea (6b)
To aniline (1eq) in water, concentrated HCl was added and the
solution was warmed. A solution of potassium thiocyanate (4eq) in
water was slowly added with stirring to above hydrochloride salt
solution. Then the solution was heated for 4-5 hrs until the solu-
tion became turbid and after cooling to room temperature the tur-
bid solution was poured in cold water. The separated precipitate
was filtered, washed with water and recrystallized from ethanol,
so as to obtain pure phenylthiourea compound (yield: 80-90%).
Thus, in view of the above factors and the recent focus of re-
searchers on 1,2,3 traizole and thiazole. In continuation of our re-
search interest on heterocyclic moieties and their biological stud-
ies [22-24], the strategy of the present study was directed towards
construction of new derivatives bearing 1,3-thiazole derived 1,2,3-
triazoles motif hybridized (Figure 1) by click chemistry reaction
via 1,3-dipolar cycloaddition of azidobenznes with alkyne.
2
.2.3. Synthesis of 4-(4-chlorophenyl)-N-phenylthiazol-2-amine (6c)
2. Experimental
A mixture of the 2-bromo-1-(4-chlorophenyl) ethanone (1eq),
1
-phenylthiourea (1eq), in the presence of K CO3 (2 eq) is stirred
2
2
.1. Materials and Methods
in DMF as a solvent for 1 hr, then the progress of the reaction is
monitored by TLC (n-hexane: EtOAc 7:3). After the completion of
the reaction, the reaction mixture is concentrate on a rotary evap-
orated under reduced pressure and the solid product was obtained
and dissolved in hot water, extracted with ether (three times x 30
All chemicals, reagents and solvents were purchased from Avra
synthesis, AR grade and were used as provided. Thin Layer Chro-
matography (TLC) analysis was performed on alumina sheets pre-
2

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
ml). The organic layer was then concentrated on a rotary evapora-
C24H18 ClN S: C, 64.93; H,4.09; N, 15.78; Found: C, 64.91; H,4.05;
5
tor under reduced pressure to furnished the desire product (6c).
N, 15.81.
4
-(4-chlorophenyl)-N-((1-(4-methyl-3-nitrophenyl)-1H-1,2,3-
triazol-4-yl)methyl)-N-phenylthiazol-2-amine (6g): NMR
1H
2
.2.4. Synthesis of 4-(4-chlorophenyl)-N-phenyl-N-(prop-2-yn-1-yl)
(
CDCl , 400MHz), δ (ppm): 8.77 (1H, s), 8.73 (1H, d, J=8Hz),
3
thiazol-2-amine (6d)
8
.5 (1H, s), 8.34-8.36 (2H, d, J=8Hz), 8.06-8.10 (3H, m), 7.98-8.02
The starting material 4-(4-chlorophenyl)-N-Phenylthiazol-2-
amine (1 eq) was added to NaH (2 eq) suspended in THF and re-
fluxed it for 20 min. then propargyl bromide(2 eq) was added and
the resulting solution was refluxed for 4 hr. When TLC of reaction
mixture showed that the starting material was completely con-
verted to product. The mixture was cooled to room temperature
and water was added. Then the solution was extracted with diethyl
ether (three times x 20 ml). Organic layer were dried over anhy-
(
(
2H, t, J=8Hz), 7.88-7.89 (3H, m), 7.26 (1H, s), 5.9 (2H, s), 2.82
3H, s); 13C NMR (CDCl , 100MHz) δ(ppm): 150.1, 138.6, 134.8,
3
1
33.5, 132.0, 132.0, 130.0, 129.1, 129.0, 123.7, 120.3, 47.3, 21.0;
+
+2
LCMS(m/z): 503(M ), 505(M ); Anal.calcd. for C25H19 ClN O S: C,
6
2
5
9.70; H.3.81; N, 16.71; Found: C, 59.68; H.3.83; N, 16.69.
-(4-chlorophenyl)-N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-
4
4
4
-yl)methyl)-N-phenylthiazol-2-amine (6h): 1H NMR (CDCl3,
00MHz), δ (ppm): 8.09 (1H, s), 7.82-7.84 (2H, d, J=8Hz), 7.61-7.63
drous Na SO filtered and evaporated to dryness to yield product.
2
4,
(
2H, d, J=8Hz), 7.42-7.51 (6H, m), 7.39-7.39 (2H, d, J=8Hz), 7.32-
Then product was purified by column chromatography in Hexane:
EtOAc (9:1) as eluent.
13
7
.34 (1H, d, J=8Hz), 6.72 (1H, s), 6.39 (2H, s); C NMR (CDCl ,
3
1
00MHz) δ(ppm): 169.2, 150.2, 145.0, 135.6, 134.6, 133.5, 133.4,
1
30.0,128.9, 127.5, 127.2, 126.2, 121.7, 121.5, 102.7, 48.2; LCMS(m/z):
+
+2
2
.2.5. General procedure for the synthesis of 1,3-thiazole derived
477.9 (M ), 479.9 (M ); Anal.calcd. for C24H17 Cl2N5S: C, 60.25; H,
3.58; N, 14.64; Found: C, 60.22; H, 3.62; N, 14.60.
1,2,3-triazoles 6(e-p)
To the stirred solution of azidobenzene derivatives 3(e-p)
1eq) and 4-(4-chlorophenyl)-N-phenyl-N-(prop-2-yn-1-yl) thiazol-
N-((1-(4-bromophenyl)-1H-1,2,3-triazol-4-yl)methyl)-4-(4-
chlorophenyl)-N-phenylthiazol-2-amine (6i): 1H NMR (CDCl3,
400MHz), δ (ppm): 8.03 (1H, s), 7.75-7.76 (2H, d, J=4Hz), 7.49-7.53
(
2
-amine (6d) (1 eq) in DCM and H O (2:1) ratio, the CuSO .5H O
2 4 2
13
(
0.3 eq) and sodium ascorbate (0.6 eq) was mixed at ambient tem-
perature. The reaction mixture was stirred at room temperature for
-6 hr. The progress of the reaction was monitored by using TLC.
(4H, d, J=16Hz), 7.19-7.41 (7H, m), 6.65 (1H, s), 5.32 (2H, s);
C
NMR (CDCl3, 100MHz) δ(ppm): 149.9, 146.5, 141.6, 134.0, 133.5,
132.0, 129.1, 127.8, 127.5, 124.2, 123.8, 120.4, 47.2; LCMS(m/z):
4
+
+2
After completion of reaction, the mixture was poured into ice cold
water (10 ml), then extracted with ethyl acetate (3 × 10 ml). The
combined organic layer was washed with water (10 ml), followed
by brine (10 ml), and dried over anhydrous sodium sulphate, con-
centrated under reduced pressure to get crude 1,3-thiazole derived
523.9 (M ), 525.9 (M ); Anal.calcd. for C24H17 BrClN5S: C, 55.13;
H, 3.28; N, 13.39; Found: C, 55.15; H, 3.25; N, 13.42.
4-(4-bromophenyl)-N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-
4-yl)methyl)-N-phenylthiazol-2-amine (6j): 1H NMR (CDCl3,
400MHz), δ (ppm): 8.11 (1H, s), 7.76-7.78 (2H, d, J=8Hz), 7.61-7.63
(2H, d, J=8Hz), 7.33-7.54 (8H, m), 7.27-7.33 (1H, d, J=16Hz), 6.74
(1H, s), 5.4 (1H, s), 5.4 (2H, s); 13C NMR (CDCl3, 100MHz) δ(ppm):
149.8, 133.7, 133.5, 132.5, 132.1, 132.1, 131.1 129.1, 123.8, 122.1,
1,2,3-triazoles 6(e-p), which were purified by column chromatog-
raphy over silica gel (60-120 mesh) using hexane: EtOAc (8:2) as
eluent.
+
+2
4
-(4-chlorophenyl)-N-Phenylthiazol-2-amine (6c): Yellow solid,
120.2, 119,3, 47.1; LCMS(m/z): 523 (M ), 525.9 (M ); Anal.calcd.
for C24H17 BrClN5S: C, 55.13; H, 3.28; N, 13.39; Found: C, 55.10; H,
3.32; N, 13.41.
1
H NMR(CDCl , 400MHz), δ(ppm): 7.75-7.78 (2H, d, J=12Hz), 7.33-
3
7
.36 (7H, m), 7.24 (1H, s), 6.79 (1H, s); 13C NMR (CDCl , 100MHz)
3
δ(ppm): 160.6, 150.2, 140.5, 134.3, 131.1, 129.3, 122.4, 117.8, 105.2;
4-(4-bromophenyl)-N-((1-(4-bromophenyl)-1H-1,2,3-triazol-
4-yl)methyl)-N-phenylthiazol-2-amine (6k): 1H NMR (CDCl3,
400MHz), δ (ppm): 8.11 (1H, s), 7.76=7.78 (2H, d, J=8Hz), 7.61-7.64
(2H, d, J=12Hz), 7.43-7.58 (9H, m), 7.31-7.34 (1H, s), 5.4 (2H, s);
+
+2
LCMS (m/z): 287(M ), 289.1(M ); Anal.calcd. for C15H11 ClN S: C,
2
6
2.82; H, 3.87; N, 9.77; Found: C, 62.86; H, 3.91; N, 9.75.
-(4-chlorophenyl)-N-phenyl-N-(prop-2-yn-1-yl)thiazol-2-
amine (6d): Pale yellow; 1H NMR(CDCl3, 400MHz), δ(ppm):
.79-7.81 (2H, d, J=8Hz), 7.43-7.51 (4H, m), 7.32-7.36 (3H, m),
.69 (1H, s), 4.80 (2H, s), 2.65 (1H. s); 13C NMR (CDCl , 100MHz)
4
13
C NMR (CDCl3, 100MHz) δ(ppm): 163.5, 161.0, 150.0, 146.9,
7
133.5, 133.3, 132.1, 129.1, 123.8, 122.3, 120.6, 120.3, 116.6, 116.3,
+
+2
6
116.1, 115.9, 47.2; LCMS(m/z): 567.9 (M ), 569.9(M ),; Anal.calcd.
for C24H17 Br2N5S: C, 50.81; H, 3.02: N, 12.17; Found: C, 50.80; H,
2.99: N, 12.19.
3
δ(ppm): 166.2, 150.2, 149.41, 134.3, 129.5, 120.8, 113.5, 105.0 78.0,
+ +2
7
3.2, 42.6; LCMS (m/z): 324 (M ), 326 (M ); Anal.calcd. for
C18 H13ClN S: C, 66.56; H, 4.03; N, 8.62; Found: C, 66.59; H, 4.00;
4-(4-chlorophenyl)-N-phenyl-N-((1-(p-tolyl)-1H-1,2,3-triazol-
4-yl)methyl)thiazol-2-amine (6l): 1H NMR (CDCl3, 400MHz), δ
(ppm): 8.09 (1H, s), 7.83-7.85 (2H, d, J=8Hz), 7.50-7.56 (4H, m),
7.42-7.46 (2H, t, J=8Hz), 7.37-7.39 (2H, d, J=8Hz), 7.27-7.30 (2H,
2
N, 8.60.
4
-(4-chlorophenyl)-N-((4-nitrophenyl)-1-H-1,2,3-triazole-4-
yl) methyl) -N-phenylthiazole-2-amine (6e): 1H NMR (CDCl3,
00MHz), δ (ppm): 8.34-8.36 (2H, d, J=8Hz), 8.22 (1H, s), 7.86-
.89 (2H, d, J=12Hz), 7.80-7.82 (2H, d, J=8Hz), 7.41-7.48 (4H, m),
13
4
m), 6.72 (1H, s), 5.41 (2H, s), 2.41 (3H, s);
C NMR (CDCl3,
7
100MHz) δ(ppm): 149.8, 146.8, 133.5, 133.4, 132.0, 130.4, 130.2,
129.1, 127.7, 125.3, 124.0, 123.8, 120.4, 47.2, 21.8; LCMS(m/z): 457.9
13
7.31-7.37 (4H,m, J=8Hz); 6.71 (1H, d, J=8Hz), 5.39 (2H, s); C NMR
+
+2
(CDCl , 100MHz) δ(ppm): 169.1, 150.2, 146.2, 144.9, 141.2, 133.5,
(M ), 459.9 (M ), Anal.calcd. for C25H20ClN5S: C, 65.56; H, 4.40;
N, 15.29; Found: C, 65.58; H, 4.38; N, 15.31.
3
130.2, 128.9, 127.6, 127.1, 126.2, 125.6, 121.4, 120.5, 102.9, 48.11;
+
+2
+3
LCMS(m/z): 489.1 (M ), 491.1 (M ), 492.1 (M ); Anal.calcd. for
C24H17 ClN O S: C, 58.95; H, 3.50; N, 17.19; Found: C, 58.96; H,
4-(4-chlorophenyl)-N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-
4-yl)methyl)-N-phenylthiazol-2-amine (6m): 1H NMR (CDCl3,
400MHz), δ (ppm): 8.01 (1H, s), 7.81-7.83 (2H, d, J=8Hz), 7.53-
7.55 (2H, d, J=8Hz); 7.48-7.50 (2H, d, J=8Hz), 7.40.7.43 (2H, t,
J=12Hz), 7.34-7.76 (2H, d, J=8Hz), 7.25-7.31 (1H, m), 6.95-6.97
6
2
3
.45; N, 17.14.
-(4-chlorophenyl)-N-Phenyl-N-((1-phenyl-1H-1,2,3- triazole-4-
yl)methyl)thiazole-2-amine (6f): 1H NMR (CDCl3, 400MHz), δ
ppm): 8.11-8.13 (1h, d, J=8Hz), 7.80-7.87(3H, m), 7.65-7.69 (2H,
4
13
(
(2H, t, J=8Hz), 6.70-6.72 (1H, s), 5.38 (2H, s), 3.82 (3H, s);
C
t, J=8Hz), 7.35-7.54 (5H, m), 7.30(2H, t, J=8Hz), 7.26 (1H, d,
NMR (CDCl3, 100MHz) δ(ppm): 169.2, 159.9, 150.2, 145.1, 133.6,
133.4, 130.6, 130.1, 128.8, 127.4, 127.2, 126.2, 121.6, 114.8, 102.7,
13
J=12Hz), 6.72 (1H, s), 5.38-5.40 (2H, s); C NMR (CDCl , 100MHz)
3
+
+2
δ(ppm): 169.3, 159.6, 150.2, 145.3, 137.1, 133.6, 131.6, 130.4, 129.8,
55.7, 48.2, 29.8; LCMS(m/z): 473.11 (M ), 475.11(M ); Anal.calcd.
for C24H17 ClFN5S: C, 63.35; H, 4.25; N, 14.78; Found: C, 63.31; H,
4.29; N, 14.81.
1
29.1,128.9, 128.5, 127.6, 126.3, 125.2, 122.6, 121.8, 120.8, 114.7,
+
+2
4
8.2, 29.8; LCMS(m/z): 444.1(M ), 446.1(M ); Anal.calcd. for
3

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
4
-(4-bromophenyl)-N-phenyl-N-((1-(p-tolyl)-1H-1,2,3-triazol-
treated groups is calculated by the equation:
1
4
-yl)methyl)thiazol-2-amine (6n): H NMR (CDCl , 400MHz), δ
3
VolumeofControl − −VolumeofTest × 100
(
ppm): 8.11 (1H, s), 7.89-7.91 (1H, d, J=8Hz), 7.76=7.78 (2H, d,
Inhibition% =
VolumeofControl
J=8Hz), 7.48-7.56 (7H, m), 7.27-7.32 (2H, m), 7.01-7.03 (2H, d,
1
3
J=8Hz), 6.90-6.94 (1H, t, J= 8Hz), 6.73 (1H, s), 3.41 (3H, s);
C
2
.3.2. In vivo anti-anxiety activity
.3.2.3. Animals. Albino rats (Wistar Strain) of either sex weighing
50-200 gram respectively were used for studies. The animals were
NMR (CDCl , 100MHz) δ(ppm): 169.5, 150.2, 144.9, 139.1, 136.6,
3
2
+
1
20.6, 119.3, 117.7, 102.9, 48.0, 21.2; LCMS(m/z): 457 (M ), 459.9
+
2), Anal.calcd. for C25H20BrN S: C, 59.76; H, 4.01, N, 13.94;
M
5
1
(
housed at room temperature (22-28 °C) with 55 ± 5% relative hu-
midity for 12 hr dark and 12 hr light cycle and given standard lab-
oratory feed (Amruth, Sangli, Maharashtra) and water ad libitum.
Found: C, 59.78; H, 3.98, N, 13.90.
4
-(4-bromophenyl)-N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-
4
4
-yl)methyl)-N-phenylthiazol-2-amine (6o): 1H NMR (CDCl3,
00MHz), δ (ppm): 7.97 (1H, s), 7.78-7.80 (2H, d, J=8Hz), 7.37-
2
.3.2.4. Experimental design of anti-anxiety activity by elevated plus
7
.45(6H, m), 7.32-7.34 (2H, d, J=8Hz), 7.25-7.29 (1H, t, J=8Hz),
13
maze apparatus. Group I: Normal group receive normal saline
6
.89-6.92 (2H, d, J=12Hz), 6.69 (1H, s), 5.38 (2H, s); C NMR
orally.
(
CDCl , 100MHz) δ(ppm): 169.4, 157.1, 150.2, 144.9, 133.5, 133.5,
3
Group II: Standard group receive Diazepam [2 mg/kg] orally.
Group III to X: Treated groups receive 5 mg/kg of compounds
orally.
1
30.1, 130, 128.8, 127.5, 127.2, 126.3, 122.3, 122.4, 121.8, 116.5,
+
+2
1
02.8, 48.15; LCMS(m/z): 506.3 (M ), 508.3 (M ); Anal.calcd. for
C24H17 BrFN S: C, 56.92; H, 3.38; N, 13.83; Found: C, 56.94; H,
5
3
.41; N, 13.79.
-(4-chlorophenyl)-N-((1-(2-nitrophenyl)-1H-1,2,3-triazol-
2
.3.2.5. Elevated plus maze. The elevated plus maze (EPM) was first
proposed as an animal model of anxiety by Handley and Mithani
27] Constructed of black–colored wooden planks consisting of two
4
-yl)methyl)-N-phenylthiazol-2-amine (6p): 1H NMR (CDCl3,
4
4
[
00MHz), δ (ppm): 8.02-8.04 (1H, t, J=12Hz), 7.79-7.81(1H, t,
close arms, 50 cm x 10 cm x 40 cm, and two open arm, 50 cm x 10
cm, connected to a central platform (10cm x 10cm). Covered with
a removable lid, such that the open or closed arms were opposite
to each other [28]. The maze was elevated to a height of 50 cm
above the floor. During the experiment each mouse was placed in
the central compartment facing one of the open arms. The number
of entries and the time spent in open arms were recorded for 5
min at 1st and 2nd hours after dosing. An entry was counted when
all four paws of the mouse entered an open or closed arm. An in-
crease in open arms entries and increase in time spent in open
arms were interpreted as an index of potential anxiolytic activity.
J=8Hz), 7.72-7.75 (1H, t, J=8Hz); 7.63-7.67(1h, t, J=8Hz); 7.56-7.58
(
1H, d, J=8Hz), 7.42-7.48 (4H, m), 7.30-7.36 (3H, m), 6.7 (1H, s),
13
5
.40 (2H, s);
C NMR (CDCl , 100MHz) δ(ppm): 150.2, 145.2,
3
1
44.9, 133.8, 133.6, 133.3, 130.7, 130.4, 130.2, 128.8, 128.0, 127.6,
+
127.2, 126.5, 125.6, 125.0, 102.8, 48.1; LCMS(m/z): 487.9 (M ),
89.9 (M+2); Anal.calcd. for C24H17 ClN O S: C, 58.95; H, 3.50; N,
6 2
4
17.19; Found: C, 58.98; H, 3.46; N, 17.15.
2
.3. Biological evaluation
2
2
.3.1. In vivo anti-inflammatory activity
.3.1.1. Animals. Wister Albino rats (180 – 250 g) were maintained
2
2
.4. Molecular Docking
in identical laboratory conditions (25°- 30°C temperature and rel-
ative humidity of 55-65 % with alternate light and darkness 12
hours each) and fed with commercial pellet diet (Hindustan Lever,
Kolkata, India) and water adlibitum [25]. This study was approved
by the Institutional Animal Ethics Committee (IAEC), B.V.V.S. Hana-
gal Shri Kumareshwar College of Pharmacy, Bagalkot-587101, Kar-
nataka, India with approval No: IAEC/HSKCOP/May 2019/MS1, Date:
.4.1. Docking procedure
The interactions of the target compounds with proteins were
analyzed using the HEX docking software. Molecular docking in-
volves the following steps [29].
• Identify a target protein
2
5 May 2019.
• Downloaded the protein PDB file and saving the PDB file
•
All the ligands were drawn in ChemDraw Ultra software and
minimum energy configuration was set for the 3D structure.
Stored the 3D optimized structure as .PDB file
2
.3.1.2. Experimental design of Carrageenan-induced rat paw edema.
•
•
•
Animals were used in the experiment have been classified into the
following three groups:
Opening both the .PDB files in HEX software
Finally running the Docking software with desired parameters
Group I: Control group receive sub plantar injection 1% suspen-
sion of carrageenan in saline and normal saline orally.
Group II: Standard group receive subplantar injection 1% sus-
pension of carrageenan in saline and Diclofenac [10 mg/kg] orally.
Group III to X: Treated groups receive sub plantar injection 1%
suspension of carrageenan in saline and 5 mg/kg of all 6e, 6g, 6h,
The parameters used for the docking process via HEX docking
software were
•
•
•
•
•
•
Correlation type – Shape only
Grid Dimension – 0.6
Receptor range – 180
Ligand Range – 180
6i, 6k, 6l and 6n compounds orally.
The in vivo anti-inflammatory of compounds (6e, 6g, 6h, 6i, 6k,
l and 6n) were investigated on carrageenan-induced inflamma-
Twist range – 360
6
Distance Range – 40
tory paw edema in mice along with standard drug delcofenac [26].
The saline (0.09%) was given to the control group at the same vol-
ume as vehicle. One hour after administration, 0.1 mL of 0.5% car-
rageen solution was injected into the footpad of the hind paws of
each mouse in all groups. Prior to carrageenan injection, the mice’
paw volume was measured with a plethysmometer. An acute in-
duced inflammation paw volume was measured at ½, 1, 3 and 5hr
since time of injection. The resultant activity of tested compounds
was compared with that of standard drug diclofenac (10mg/kg)
orally. The percentage inhibition of paw edema volume of each
3. Result and Discussion
3.1. Chemistry
Generally, 1,2,3-triazole pharmacophores moiety can be syn-
thesized by the Cu(I)-catalyzed 1,3-dipolar cycloaddition reaction.
The most prominent example of click chemistry is the Cu(I)-
catalyzed azide-alkyne click chemistry (CuAAC) reaction [30]. An
4

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
Scheme 1. General reaction for synthesis of triazole moiety
Scheme 2. Reaction pathway for the synthesis of 1,3 thiazoles derived 1,2,3-traizoles (6c-p).
Reagents and conditions: (i) K2CO3, DMF (ii) Propargyl bromide, K2CO3, DMF, N2 atmosphere, 60°C, 24 hr; (iii) NaNO2, Conc. HCl, 0-5°C 15min; (iv) Aq. NaN3 (v)
AcCN/H2O = 2:1, corresponding azide, 1 M CuSO4.6H2O solution, 1M Sodium Ascorbate, 60°C 8 hr;
azide-functionalized molecule (A) reacts with a terminal Alkyne-
functionalized molecule (B) thereby forming a stable conjugate A-B
via a triazole moiety (Scheme 1).
of 5mg/kg as compared with control group, which proves their po-
tent anti-inflammatory activity. The percentage inhibition of edema
increased in acute inflammation and paw volume was significantly
reduced by target compounds (6e, 6g, 6h, 6i, 6k, 6l, and 6n) at
½, 1, 3 and 5 hr and the % inhibition of edema by target com-
pounds was statistically similar to that of standard drug diclofenac
(Table 2, Figure 2 and 3). They exhibited an excellent reduction in
edema paw volume at first half hour of treatment, which is closer
to the value found for the standard drug used as positive control.
Generally, literature review revealed that the, electron withdrawing
groups are favours for the increasing in the anti-inflammatory ac-
tivity of synthesized compounds [31]. This might be the region the
compounds (6g, 6h 6i and 6k) having Cl, di-NO2 and Br groups on
the phenyl ring showed potent ant inflammatory activity. The com-
pounds (6l and 6n) having CH3 and OH groups showed moderate
to good activity compared to the compounds (6g, 6h and 6k). This
clearly indicates that the compounds with electron withdrawing
The synthetic approaches of contraction of the target 1,3-
thiazole derived 1,2,3-traizole hybrids 6(e-p) (Scheme 2) and de-
tailed was under the experimental section. Initially, the azidoben-
zene derivatives have been synthesized by diazotization of sub-
stituted aniline 1(e-p) and finally converting resultant aryl dia-
zonium salt 2(e-p) into azidobenzene derivatives 3(e-p) by us-
ing sodium azide. The intermediate compound (6c) was afforded
by the reaction of 2-bromo-1-(4-chloro/Bromo phenyl) ethanone
and 1-phenyl thiourea gives 4-(4-chlorophenyl)-N-phenylthiazol-
2
-amine (6c). In the next, the key scaffold (6d) was ob-
tained by base induced reaction between 4-(4-chlorophenyl)-N-
phenylthiazol-2-amine (6c) with propargyl bromide. Finally 1,3
thiazoles derived 1,2,3-traizoles was furnished by reaction of 4-
(
4-chlorophenyl)-N-phenyl-N-(prop-2-yn-1-yl)thiazol-2-amine (6d)
and azidobenzene derivatives 1(e-p) by copper catalysed cycload-
dition reaction offered N,4-diphenyl-N-((1-phenyl-1H-1,2,3-triazol-
groups (Cl, NO ) on the phenyl ring are better anti-inflammatory
2
agents [32] and these groups were also found in NSAIDs such as di-
clofenac, indomethacin, ketoprofen, meclofenamic and nimesulide
(selective for COX-2) [33].
4
-yl)methyl)thiazol-2-amine derivatives 6(e-p) with an good yield
of 65-96% (Table 1). All the compounds 6(c-p) were characterized
properly by 1H NMR, 13C NMR, LCMS and elementary analysis. The
1H NMR spectra of compounds 6(e-p) containing aliphatic (-2H-)
protons are characterized by the presence of signal at high field
3.2.2. In vivo anti-anxiety activity
The investigation of anti-anxiety activity of target compounds
(6e, 6g, 6h, 6i, 6k, 6l, and 6n) on albino mice have contributed
to therapeutic applications. These target molecules associated with
1,2,3-triazole and thaizole have the potential for anxiety. In this
study, we have used EPM model of anxiety to evaluating an anx-
iolytic effect of target compounds. As expected, diazepam gives
significance increased in time spend and number of entries into
open arm and sat same time decrease in time spent and number
of entries in closed arm of EPM model. The behavioural changes
induced by compounds are consistent with an anxiolytic effect,
and it is also statistically similar to that of diazepam. Some tar-
get compounds have shown significant anti anxiety activity and
the substituent’s present on the phenyl ring of thiazole and tria-
zole may be playing a vital role in enhanced anti anxiety of tested
5
3
3
.34-5.98ppm.
.2. Biological evaluation
.2.1. In vivo anti-inflammatory activity
The objective of this study is to screen in vivo anti-
inflammatory activity of compounds (6e-p) and compare with
treated control and diclofenac used as a standard drug . The re-
sults were summarized in table 3 which indicated that the con-
trol group showed increase in paw volume [ml] from half hour
to fifth hour and anti-inflammatory activity of synthesized com-
pounds showed a significant inhibition [p<0.05 to p<0.001] of
edema by decreasing paw volume in experimental rats at a dose
5

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
Table 1
compounds. The earlier structure-activity relationship studies that
compounds having electron withdrawing substituent like chloro,
bromo, nitro, triflouromethyl and cyano groups at phenyl ring,
small polar/hydrophilic substitutions, and chloro, bromo or flouro
substitutions at different position of phenyl ring of the heterocyclic
ring skeleton may enhance anti-anxiety activity [34, 35]. Com-
pound 6i bearing chloro group on thaizole substituted phenyl ring
and bromo group on triazole substituted phenyl ring has shown
the highest anti anxiety activity compared to other target com-
pounds (Figure 4 and 5). Moreover in silico molecular docking was
predicted with docking score -268.36 (6i), which is comparatively
near to the reference drug -285.12 (Diazepam). The ligand bind-
ing interaction diagram reveals the similar binding interaction with
key amino acid residues. Such interactions may have stabilized the
test compounds insides the GABAA receptor binding pocket and to-
tal docking score predicted to be a comparable to before diazepam.
However, the compounds 6e and 6h containing Cl and NO2 groups
on the substituted phenyl rings also produced statistically signifi-
cant anti anxiety activity. Whereas, rest of the compounds 6g, 6k,
Chemical structure, yield and melting point of synthesized compounds 6(c-p).
Sl. No
Structure of Compound
M. Pt
Yield (%)
6
6
c
177-179
72
d
184-186
80
6
6
e
f
252-254
168-170
78
62
6l, and 6n showed moderate activity. The obtained result were ex-
pressed as a Mean ± SEM and summarized in Table 3 and 4.
3
.2.3. Acute toxicity study (LD50
)
The acute toxicity of synthesized compounds was studied ac-
6
6
6
6
6
g
h
i
176-178
190-192
193-195
200-202
208-210
65
cording to the OECD guidelines 425 and the female Swiss albino
mice (25-30 gm) were used for acute toxicity. Limit dose of 2000
mg/kg (p.o.) was administered and followed by behavioural and
toxicological signs were observed for 4 h and monitored up to 14
days and mortality was not occurred with higher limit dose. LD50
was calculated accordingly and found more than 2000 mg/kg. The
same has been included in the manuscript.
77
82
3
.3. Molecular Docking
The results obtained by experimentation of anti-anxiety activ-
ity were further supported by the outcomes of molecular dock-
ing studies. The tested compounds(6e, 6g, 6h, 6i, 6k, 6l, and 6n)
were found to possess tremendous affinity towards GABAA recep-
tors and may therefore be deemed as imperative targets to develop
potential anti anxiety agents. All the test compounds were furth er
docked and their docking scores were predicted in Table 5. The
new 1,3 thiazoles derived 1,2,3 traizole disclosed the diverse note-
worthy interaction with active site of amino acid radius (Ile107;
ala122; Asp121; Lys66; Val63; Lys74; Hoh6020; Arg85; Asn67;
Cys65; Thr64; Cys72; Ser123; Glu61). The scaffolds with elec-
tron withdrawing groups (EWGs) particularly Cl & Br on the tri-
azole and thaizole substituted phenyl rings, exhibited good inter-
actions with amino acids residues and posses the highest dock-
ing energy. Molecular docking studies provided an interaction map
of GABAA with tested compounds. The 1,2,3-traizole derived 1,3-
thiazoles compounds (6h) and (6i) exhibits four H-bond interac-
tion represent in Figure 6, 7 and 8, from that two stronger bond
observed with Asp-121 and Ala122, in the other hand compound
j
80
k
75.5
6
l
196-198
68
6
6
m
152-154
166-168
55
78
(
6e) exhibits two hydrogen bond with Asp-831 and Ala-122. Over-
all 1,2,3 traizole derived 1,3 thiazoles derivatives posses signifi-
cant H-bond interaction with the notable interaction like Pi-Pi-
stacking (dark pink), alkyl (light pink), pi-alkyl (pale pink), Pi-
sulfur (yellow), Pi-lone pair (light green) and pi-anion (orange) in-
teraction were also found [36]. Based on the docking energy, the
test compounds are being more effective inhibitors in following or-
der 6i>6h>6e>6g>6l>6j>6n>6k. Molecular docking is regularly
used for understanding the drug protein receptor interaction with
modern drug design field.
n
6
6
o
p
210-212
155-157
72
65
6

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
Figure 2. Anti-inflammatory effect of tested compounds on carrageenan induced acute inflammation in rats. All values were expressed as a Mean ± SEM. One Way Analysis
∗
∗∗
∗∗ ∗
p<0.001, p<0.01, p<0.05 as compared to control groups.
of Variance (ANOVA), followed by multiple comparison Dunnet’s test. The p value
Figure 3. Anti-inflammatory effect of tested compounds on carrageenan induced acute inflammation in rats. All values were expressed as a Mean ± SEM. One Way Analysis
∗
∗∗
∗∗ ∗
p<0.001, p<0.01, p<0.05 as compared to control groups.
of Variance (ANOVA), followed by multiple comparison Dunnet’s test. The p value
Figure 4. Anti-anxiety effect of tested compounds on number of entries in open arm in elevated plus maze anxiety model. All values were expressed as a Mean ± SEM. One
∗
∗∗
∗∗ ∗
p<0.001, p<0.01, p<0.05 as compared to control groups.
Way Analysis of Variance (ANOVA), followed by multiple comparison Dunnet’s test. The p value
Figure 5. Anti-anxiety effect of synthetic 6 series compounds on time spent in open arm in elevated plus maze anxiety model. All values were expressed as a Mean ± SEM.
∗
∗∗
∗∗ ∗
p<0.001, p<0.01, p<0.05 as compared to control groups.
One Way Analysis of Variance (ANOVA), followed by multiple comparison Dunnet’s test. The p value
7

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
Table 2
Effect of target tested compounds on carrageenan induced acute inflammation in rats.
Treated Groups
Paw edema volume in ml (% of edema inhibition)
1/2hr
1hr
3hr
5hr
Control
0.7433± 0
0.6600± 0
0.5800± 0.1206
0.4967± 0.07860
(79.20%)
0.8766± 0.0002963
∗
∗∗
∗∗∗
∗∗
∗∗
∗∗∗
Diclofenac
0.100± 0.06009
0.123± 0.1418
(81.31%)
0.4533± 0.04702
(
86.54%)
(48.29%)
∗∗∗
∗∗∗
6
5
6
5
6
5
6
5
6
5
6
5
6
5
e
0.2467± 0.06984
0.3400± 0.1739
(48.48%)
0.1767± 0.03844
0.4167± 0.05457
mg/kg
(66.81%)
(69.53%)
(52.46%)
∗
∗∗
0.2167± 0.06333∗
∗∗∗
g
0.2433± 0.1041
0.2100± 0.1305
(68.1%)
0.3767± 0.008819
mg/kg
(67.26%)
(62.63%)
(57.07%)
∗
∗
∗
∗
∗∗
∗∗
∗∗∗
h
0.1567± 0.03844
(78.91%)
0.3033± 0.1572
(54.04%)
0.1533± 0.09770
(73.56%)
0.3400± 0.0200
mg/kg
(61.21%)
∗
∗∗∗
i
0.3533± 0.09597
(52.46%)
0.5200± 0.1484
0.6633± 0.1241
0.5100± 0.08737
mg/kg
k
(21.21%)
(-14.36%)
(41.82%)
∗∗
∗∗
0.1633± 0.05897∗
(75.25%)
0.2300± 0.07211∗
(60.03%)
∗∗∗
0.1000± 0.06245
(86.54%)
0.3633± 0.008819
mg/kg
l
(58.55%)
0.1000± 0.06245∗
0.1167± 0.01856
(96.8%)
∗∗
0.4933± 0.2478
∗∗∗
0.2433± 0.09207
mg/kg
n
(67.26%)
(84.84%)
(43.72%)
∗
∗∗
0.1233± 0.03756∗
(81.31%)
∗∗
∗∗∗
0.067± 0.02963
0.1800± 0.05033
(91.32%)
0.6367± 0.1419
mg/kg
(91.03%)
(27.37%)
All values were expressed as a Mean ± SEM. One Way Analysis of Variance (ANOVA), followed by multiple comparison Dunnet’s test. The
∗
∗∗
∗∗ ∗
p<0.001, p<0.01, p<0.05 as compared to control groups.
p value
Figure 6. 3D and 2D Interactions of compound (6i) with protein receptor (PDB
code:1KJT Crystal Structure of the GABA(A)
Figure 7. 3D and 2D Interactions of compound (6e) with protein receptor (PDB
code:1KJT Crystal Structure of the GABA(A)
Table 3
Anti-anxiety effect of tested compounds on number of entries in
open arm in elevated plus maze anxiety model.
Table 4
Anti-anxiety effect of tested compounds on time spent in open arm in elevated
plus maze anxiety model.
Treated Groups
NO. OF ENTRIES IN OPEN ARM
Treated Groups
TIME SPENT IN OPEN ARM [sec]
2nd hr
1st hr
2nd hr
1st hr
Normal
0.5000± 0.2236
0.3333± 0.2108
∗
∗
∗
∗
∗
∗∗
∗∗
∗∗∗
∗∗∗
∗∗
∗∗∗
∗∗∗
Diazepam 2mg/kg
8.833± 0.6540
3.667± 0.3333
2.833± 0.4014
4.833± 0.6009
7.000± 0.5774
1.500± 0.2236
2.333± 0.4944
2.167± 0.3073
2.167± 0.6009
9.500± 0.3416
4.167± 0.4773
3.500± 0.6191
5.333± 0.6667
6.667± 0.8433
1.333± 0.2108
1.833± 0.4014
1.833± 0.3073
1.667± 0.4944
Normal
7.000± 0.5774
7.833± 0.6540
∗∗∗
6
6
6
6
6
6
6
6
e
g
h
i
Diazepam 2mg/kg
138.3± 8.301
48.00± 7.109
33.50± 5.638
90.33± 3.029
∗∗∗
160.0± 13.45
51.33± 7.592
35.33± 5.239
94.17± 4.672
∗∗∗
∗
∗∗∗
∗∗∗
∗
∗∗∗
6e
6g
6h
6i
∗∗
∗∗
j
116.5± 2.693∗∗∗
118.3± 2.704∗∗∗
k
l
6j
18.67± 3.547
15.67± 2.044
20.33± 5.057
22.00± 5.428
20.50± 4.372
17.67± 2.108
22.17± 4.778
20.50± 5.071
6k
6l
n
6
n
All values were expressed as a Mean ± SEM. One Way Analysis
of Variance (ANOVA), followed by multiple comparison Dunnet’s
All values were expressed as
a
Mean
±
SEM. One Way Analysis of Vari-
∗
∗∗
∗∗
∗
test. The p value
p<0.001, p<0.01, p<0.05 as compared to
ance (ANOVA), followed by multiple comparison Dunnet’s test. The p value
∗∗∗p<0.001, ∗∗p<0.01, ∗p<0.05 as compared to control groups.
control groups.
8

K.N. Ankali, J. Rangaswamy, M. Shalavadi et al.
Journal of Molecular Structure 1236 (2021) 130357
Table 5
The docking energy and hydrogen bond interaction with amino residue of tested compounds.
Compounds
Docking energy in (Kcal/mol)
Docker energy
Hydrogen bonding residues
Diazepam
-285.12
-243.11
-220.52
-262.25
-268.36
-198.15
-192.31
-210.53
-198.10
6
6
6
6
6
6
6
6
e
g
h
i
Val-124, Arg-85, Val-43, Ala-122, Ile-107, Lys-74, Val-63, Thr-64, Tyr-73, Lys-66, Cys-65,74, The-120, Asp-121, Asn-67
Lys-74, Ile-107, Ala-122, Asp-121, Val-63, Lys-66
Ile-107, Ala-122, Asp-121, Lys-62, Cys-74, Val-63 Thr-64, Lys-66,
Glu-61, Lys-74, Ile-107, Ala-122, Asp-121, Lys-62, Cys-74, Val-63 Thr-64, Lys-66, Asn-67
Ala-122, Asp-121, Val-63, Lys-66, Ile-107,
j
k
l
Lys-74, Ile-107, Ala-122, Asp-121, Lys-62, Cys-74, Val-63 Thr-64, Asn-67
Val-43, Ile-107, Lys-74, Val-63, Thr-64, Tyr-73, Asp-121, Ala-122
n
Lys-74, Val-63, Thr-64, Tyr-73, Asp-121, Ala-122, Asp-121, Val-43, Ile-107
Declaration of competing interest
No conflict of interest.
Acknowledgement
Prof. N.N. would like to acknowledge UGC-SAP-DRS-III (UGC F
5
40/10/DRS-III/2016 (SAP-I)), “K. N. A. would like thanks to CSIR-
NET-JRF (09/119(0208)/2018-EMR-I” and J. R. thanks to UGC-PDFSS
UGC Order No. F./PDFSS-2015-17-KAR-11614) for their financial
(
support in the present work.
Supplementary materials
Supplementary material associated with this article can be
found, in the online version, at doi:10.1016/j.molstruc.2021.130357.
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