Design, synthesis, anticancer activity and molecular docking analysis of novel dinitrophenylpyrazole bearing 1,2,3-triazoles

Article abstract of DOI:10.1016/j.molstruc.2021.130865

A novel series of dinitrophenylpyrazole bearing triazole scaffolds were designed and synthesized with appreciable yields. The structures of all the target molecules (9a–j) were confirmed and characterized by ¹H NMR, ¹³C NMR, FT-IR and HR-MS spectral techniques. These novel molecules were tested for their anti-cancer activity (in vitro) using three tumor cell lines indicating breast adenocarcinoma (MCF-7), cervical carcinoma (HeLa), and human epithelial colorectal adenocarcinoma (Caco-2) via the MTT approach. Compounds 9e, 9f, and 9 h showed the high potency growth inhibitory activity against the HeLa cell line (IC₅₀ = 4.0 μM, 5.0 μM and 6.0 μM) higher than that of Combretastatin-A4 (IC₅₀ = 9.0 μM) as reference drug. Besides, compound 9 h was found to be highly potent against MCF-7 cell line IC₅₀ value of 8.0 μM. Additionally, in this manuscript, the structure-activity relationship (SAR) analysis is reported. Molecular docking studies also confirm the experimental findings and explains the most probable interaction pattern.

Full text of DOI:10.1016/j.molstruc.2021.130865

Journal of Molecular Structure 1243 (2021) 130865
Contents lists available at ScienceDirect
Journal of Molecular Structure
journal homepage: www.elsevier.com/locate/molstr
Design, synthesis, anticancer activity and molecular docking analysis
of novel dinitrophenylpyrazole bearing 1,2,3-triazoles
Kotyada Suryanarayanaa, Alice Rinky Roberta, Nagaraju Kerruc, Thangaiyan Pooventhirand,
Renjith Thomasd, Suresh Maddilaa,b,∗, Sreekantha B. Jonnalagaddab
a Department of Chemistry, GITAM Institute of Sciences, GITAM University, Visakhapatnam, Andhra Pradesh, India
b School of Chemistry and Physics, University of KwaZulu-Natal, Westville Campus, Chiltern Hills, Durban 4000, South Africa
c Department of Chemistry, GITAM School of Science, Bengaluru Campus, Karnataka 561203, India
d Department of Chemistry, St Berchmans College, Mahatma Gandhi University, Changanassery, Kerala, India
a r t i c l e i n f o
a b s t r a c t
Article history:
A novel series of dinitrophenylpyrazole bearing triazole scaffolds were designed and synthesized with
appreciable yields. The structures of all the target molecules (9a–j) were confirmed and characterized by
1H NMR, 13 C NMR, FT-IR and HR-MS spectral techniques. These novel molecules were tested for their
anti-cancer activity (in vitro) using three tumor cell lines indicating breast adenocarcinoma (MCF-7), cer-
vical carcinoma (HeLa), and human epithelial colorectal adenocarcinoma (Caco-2) via the MTT approach.
Compounds 9e, 9f, and 9 h showed the high potency growth inhibitory activity against the HeLa cell line
(IC50 = 4.0 μM, 5.0 μM and 6.0 μM) higher than that of Combretastatin-A4 (IC50 = 9.0 μM) as reference
drug. Besides, compound 9 h was found to be highly potent against MCF-7 cell line IC50 value of 8.0 μM.
Additionally, in this manuscript, the structure-activity relationship (SAR) analysis is reported. Molecular
docking studies also confirm the experimental findings and explains the most probable interaction pat-
tern.
Received 27 March 2021
Revised 27 May 2021
Accepted 7 June 2021
Available online 11 June 2021
Keywords:
Synthesis
Pyrazoles
Triazoles
Cytotoxic activity
HeLa
Docking analysis
© 2021 Elsevier B.V. All rights reserved.
1. Introduction
site and are used as positive controls for the tubulin depolymer-
ization assay [7]. However, the survival of peripheral neuropathy is
Cancer is considered one of the most widespread health hitches
to human beings in all developed countries and is one of the most
leading causes of death [1,2]. The discovery of novel therapies is a
principal area of significant medicinal and pharmaceutical research
[3]. Several cancers such as breast, colon, lung, and melanoma are
the most prevalent in developing and underdeveloped countries
[4]. Scientists have primarily been perturbing to discover effective
clinical investigations for tumor treatment and are in pursuit of
new anti-cancer agents for many decades [5]. Microtubules, poised
of α,β-tubulin heterodimers, are key elements in the eukaryotic
cells of the cytoskeleton. Which are pleiotropic in their role, mainly
in coordinating the spatial dissemination of organelles in chromo-
somes and cells during cell division. Owing to their vital positions
in the cell, microtubules provide a desirable drug target [6]. The
ligand binding of microtubules acquires three sites viz., colchicine,
taxan, and vinca domain. Combretastatin A-4 and colchicine are
key tubulin-binding agents by binding at the colchicine binding
a general constraint in the growth of anti-microtubule molecules
as drugs [8]. Therefore, there is a necessity to explore and develop
new anti-cancer medicinal compounds that are efficient in pre-
venting proliferation and neuropathies [9]. Anti-cancer drugs ex-
hibit a crucial role in cancer treatment, whether utilized unaided
and/or coupled with other treatments like irradiation, surgery, and
chemotherapy [10]. Currently, chemotherapy amalgamation using
drugs with various mechanisms of action is being advocated to
cure cancer, thereby diminishing the toxicity of the therapy. Conse-
quently, a single compound having more than one pharmacophore
structure, each with diverse kinds of activity, can be valuable for
cancer therapy.
Among heterocyclic compounds, nitrogen comprising heterocy-
cles have preserved the importance of scientists and academicians
and are key building blocks applied for the advance of a variety of
biological and medical important skeletons [11–13]. Their distinc-
tive constructions have led to various applications in several areas
such as medicinal, pharmaceutical, industrial, computational, and
agricultural fields [14]. Synthesis of these heterocycles has been
among the major research areas in organic chemistry based on the
inclination towards heterocyclic molecules [15]. Pyrazoles are no-
∗
Corresponding author at: Department of Chemistry, GITAM Institute of Sciences,
GITAM University, Visakhapatnam, Andhra Pradesh, India.
E-mail address: sureshmskt@gmail.com (S. Maddila).
https://doi.org/10.1016/j.molstruc.2021.130865
0022-2860/© 2021 Elsevier B.V. All rights reserved.

K. Suryanarayana, A.R. Robert, N. Kerru et al.
Journal of Molecular Structure 1243 (2021) 130865
table examples of aromatic heterocycles containing two nitrogen
atoms in a five-membered ring and are important pharmacophores
in anticancer research [16]. These are very flexible lead compounds
to synthesize strong bioactive moieties for drug revelation and ad-
vancement, especially in malignancy treatment [17]. Pyrazoles and
their derivatives reveal important biological properties like anti-
bacterial, anti-microbial, anti-neoplastic, anti-inflammatory, anti-
convulsant, anti-diabetic, anti-spasmodic, anti-hyperglycemic, anti-
depressive, anti-cancer, analgesic, antipyretic, and anti-malarial ac-
tivities [18–21]. Further, they have been broadly utilized in her-
bicides and pesticides [22,23]. In addition, 2,3-diphenylcycloalkyl
pyrazole and pyrazole-naphthalene scaffolds proved to have potent
anticancer action due to the inhibition of tubulin polymerization
and arrested the cells in the G2/M phase of the cell cycle [24,25].
Some, pyrazole-arylcinnamide derivatives showed a better growth
inhibition impact against HeLa cell lines apart from tubulin poly-
merization inhibitory activity [26]. Among the different FDA per-
mitted therapeutic medicines, pyrazole moiety is a component of
Celebrex, Sildenafil, and Rimonabant drugs [27].
2.1.2. General procedure for the synthesis of compound (4)
To a solution of compound (3) (0.01 mol) in DMF (6 mL), phos-
phoryl chloride (POCl3, 4 mL) was gradually added in a dropwise
manner. The resulting mixture was stirred at 60 °C for about 5 h.
Then, the solvent was evaporated under reduced pressure, and the
resulting mixture was neutralized by adding an aqueous NaOH so-
lution. Further, the crude was filtered, washed with water, dried,
and recrystallized from ethanol.
2.1.3. General procedure for the synthesis of compound (6)
A mixture of equimolar amounts of substituted pyrazole-4-
carbaldehydes (4) (0.01 mol) and para-amino phenol (5) (0.01 mol)
in ethanol solvent (10 mL) was refluxed for 2–3 h. The reaction
progress was observed using TLC at different time intervals. After
completion of the reaction, the solvent was evaporated under re-
duced pressure until the solid separated. Finally, the formed crude
was purified using column chromatography.
2.1.4. General procedure for the synthesis of compound (7)
1,2,3-triazole is a significant class of heterocyclic scaffold con-
sisting of a five-membered ring with three nitrogen atoms [28,29].
It has a large variety of medicinal applications. In several het-
erocyclic drugs, 1,2,3-triazoles are the central core. They have
proven outstanding biological and pharmacological activities like
anti-malarial, anti-convulsant, anti-bacterial, anti-tumor, anti-viral,
anti-inflammatory, anti-cancer, analgesic, antioxidant agents [30–
33]. These varied features of the 1,2,3-triazole moiety have exhila-
rated investigators to design new triazole molecules with improved
potential. Due to their biological and pharmacological attributes,
they have gained prominence in their synthesis.
A mixture of compound (6) (0.01 mol) and propargyl bromide
(0.015) were made to react in the presence of potassium carbon-
ate (0.03 mol) in dimethylformamide solvent for 2 h at R.T. After
the reaction is completed; the reaction mixture was poured into
crushed ice with continuous stirring. The solid mass thus separated
was filtered, dried, and purified by using diethyl ether.
2.1.5. General procedure for the synthesis of pyrazole-triazole
derivatives (9a–j)
To a stirred solution of compound (7) in DCM:H2O (1:1), dif-
ferent substituted azides (8 a–j; 2 mmol) were added, followed
by the addition of copper sulfate (0.055 mmol) and sodium ascor-
bate (0.13 mmol) at room temperature. The progress of the reac-
tion was monitored using TLC until the disappearance of starting
material. Then water (15 mL) was added to the reaction mixture
and extracted with chloroform (50 mL). Combined organic layers
were dried over anhydrous sodium sulfate and concentrated un-
der reduced pressure resulting in the isolation of a crude prod-
uct, which was purified by silica gel chromatography using hex-
ane:ethylacetate (60:40) to get the pure target products. The struc-
tural interpretation of novel molecules was made using various
spectroscopic approaches (1H NMR, 13C NMR and HR-MS) and in-
strumental specifics provided in the supporting information (SI-1
and SI-2).
Our previous work has reported the novel synthesis of vari-
ous fused heterocyclic molecules with different biological activi-
ties like anti-microbial, anti-inflammatory, antioxidant, and anti-
cancer evaluation [34–39]. We have recently developed facile,
environmental-friendly novel heterogeneous catalysts to synthe-
size various heterocyclic molecules under green conditions [40–
44]. The versatile skeletons of microtubule-binding molecules, in
divergence to new types of anticancer agents, is their immense
structural diversity and complexity, which gives various prospects
for the progress of new scaffold design. However, the pyrazole-
based analogs can be developed further to produce better anti-
tubulin scaffolds that could involve tubulin polymerization. En-
couraged with the facts mentioned above and in continuation of
our research studies in this area, we envisioned the combination
of dinitrophenylpyrazole pharmacophore unit and 1,2,3-triazole hy-
brid framework for anti-cancer activity and molecular docking
analysis. To the best of our knowledge, there are no reports on the
synthesis of dinitrophenylpyrazole bearing 1,2,3-triazole scaffolds
along with their anti-cancer activity or molecular docking analysis.
2.1.5.1. 4-((1-(2,4-dichlorophenyl)−1H-1,2,3-triazol-4-yl)methoxy)-N-
((1-(2,4-dinitro phenyl)−3-phenyl-1H-pyrazol-4-yl)methylene)aniline
(9a). FT-IR νmax (cm−1): 3130 (=C H), 2838 (C–H), 1612 (C = C),
–
1532 (C = N); 1333 (NO2); 1H NMR (400 MHz, DMSO–d6) δ 9.86
(s, 1H, CH-triazole), 9.38 (s, 1H, CH), 8.97 (s, 1H, Ar-H), 8.91 (dd,
J = 6.6, 2.5 Hz, 1H, Ar-H), 8.71 (m, 3H, Ar-H), 8.40 (d, J = 8.9 Hz,
2H, Ar-H), 8.32 (d, J = 8.9 Hz, 1H, Ar-H), 7.86 (d, J = 1.8 Hz, 2H,
Ar-H), 7.78 (d, J = 1.8 Hz, 1H, Ar-H), 7.62 (dd, J = 8.3, 1.9 Hz, 1H,
Ar-H), 7.57 (dd, J = 8.2, 2.0 Hz, 1H, Ar-H), 7.46 (d, J = 8.3 Hz, 2H,
Ar-H), 7.00 (s, 1H, CH), 5.26 (s, 2H, CH2); 13C NMR (100 MHz,
DMSO–d6) δ 168.89, 152.31, 150.72, 146.61, 146.34, 143.02, 137.50,
135.58, 135.08, 135.06, 133.75, 131.92, 129.66, 129.25, 128.32,
128.04, 127.95, 127.52, 127.29, 127.08, 126.15, 123.61, 121.28, 121.16,
2. Experimental section
2.1. Chemistry
2.1.1. General procedure for the synthesis of compound (3)
To
a solution of acetophenone, compound (1) (0.01 mol)
taken in 10 mL ethanol was added 2,4-dinitrophenyl hydrazine
(0.02 mol), and the reaction mixture was stirred for 30 min, and
sulphuric acid (1 mL) was then added. The reaction mixture was
stirred continuously at room temperature for 1 h until found com-
plete as monitored by TLC. On completion of the reaction, the mix-
ture was poured into cold water, and the crude solid formed was
filtered off, washed, dried, and finally recrystallized from ethanol
to obtain 1-(2,4-dinitrophenyl)−2-(1-phenylethylidene)- hydrazine
(3).
119.81, 117.90, 71.97; HRMS of [C31H20N8O5Cl2
677.2083; Calcd: 677.2080.
+
Na]+ (m/z)
2.1.5.2. 4-((1-(4-chlorophenyl)−1H-1,2,3-triazol-4-yl)methoxy)-N-((1-
(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-yl)methylene)aniline (9b).
FT-IR νmax (cm−1): 3013 (=C H), 2956 (C–H), 1604 (C = C), 1509
–
(C = N); 1334 (NO2); 1H NMR (400 MHz, DMSO–d6) δ 8.93 (s,
1H, CH-triazole), 8.92 (s, 1H, CH), 8.69 (dd, J = 8.9, 2.5 Hz, 1H,
Ar-H), 8.39 (d, J = 8.9 Hz, 1H, Ar-H), 8.22 (dd, J = 16.4, 5.9 Hz,
2

K. Suryanarayana, A.R. Robert, N. Kerru et al.
Journal of Molecular Structure 1243 (2021) 130865
4H, Ar-H), 7.88 (d, J = 7.3 Hz, 1H, Ar-H), 7.66 – 7.44 (m, 9H, Ar-H),
7.37 (s, 1H, CH), 5.12 (s, 2H, CH2); 13C NMR (100 MHz, DMSO–d6)
δ 168.91, 155.07, 146.11, 142.83, 135.14, 132.09, 130.24, 129.56,
129.50, 129.24, 129.11, 128.95, 128.93, 128.74, 128.64, 128.57,
128.53, 128.39, 127.96, 126.90, 126.31, 121.16, 120.46, 116.73, 78.93;
HRMS of [C31H21N8O5Cl + Na]+ (m/z) 643.2527; Calcd: 643.2533.
124.84, 120.92, 120.43, 118.98, 118.12, 114.98, 112.16, 112.10, 110.73,
71.97, 55.53; HRMS of [C32H24N8O6 + H]+ (m/z) 617.2519; Calcd:
617.2523.
2.1.5.7. N-((1-(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-
yl)methylene)−4-((1-phenyl-1H-1,2,3-triazol-4-yl)methoxy)aniline
(9 g). FT-IR νmax (cm−1): 3054 (=C H), 2839 (C–H), 1582 (C = C),
–
2.1.5.3. 4-((1-(4-bromophenyl)−1H-1,2,3-triazol-4-yl)methoxy)-N-((1-
(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-yl)methylene)aniline (9c).
1501 (C = N); 1322 (NO2); 1H NMR (400 MHz, DMSO–d6) δ 8.98
(s, 1H, CH-triazole), 8.92 (s, 1H, CH), 8.70 (dd, J = 8.9, 2.4 Hz,
1H, Ar-H), 8.41 (d, J = 8.9 Hz, 1H, Ar-H), 8.31 (d, J = 3.9 Hz, 2H,
Ar-H), 7.70 (d, J = 7.9 Hz, 1H, Ar-H), 7.71–7.62 (m, 5H, Ar-H), 7.56
– 7.48 (m, 5H, Ar-H), 7.45 (d, J = 7.4 Hz, 2H, Ar-H), 6.98 (s, 1H,
CH), 5.24 (s, 2H, CH2); 13C NMR (100 MHz, DMSO–d6) δ 167.89,
165.81, 164.43, 153.85, 149.77, 146.38, 143.01, 141.56, 138.68,
135.04, 130.31, 129.63, 129.24, 129.09, 128.64, 128.58, 128.36,
128.19, 126.58, 122.87, 121.22, 119.72, 117.94, 115.11, 71.73; HRMS
of [C31H22N8O5 + Na]+ (m/z) 609.2761; Calcd: 609.2766.
FT-IR νmax (cm−1): 3065 (=C H), 2839 (C–H), 1638 (C = C), 1532
–
(C = N); 1322 (NO2); 1H NMR (400 MHz, DMSO–d6) δ 8.92 (s, 1H,
CH-triazole), 8.87 (s, 1H, CH), 8.67 (dd, J = 17.8, 11.1 Hz, 1H, Ar-H),
8.39 (d, J = 8.9 Hz, 1H, Ar-H), 8.24 (d, J = 9.0 Hz, 2H, Ar-H), 8.19
(d, J = 9.0 Hz, 2H, Ar-H), 7.85 (d, J = 8.2 Hz, 2H, Ar-H), 7.70 – 7.55
(m, 5H, Ar-H), 7.42 (t, J = 8.7 Hz, 1H, Ar-H), 7.35 (s, 1H, CH), 7.19
(d, J = 2.4 Hz, 2H), 5.45 (s, 2H, CH2); 13C NMR (100 MHz, DMSO–
d6) δ 168.86, 164.30, 164.10, 154.08, 146.14, 145.01, 142.82, 135.10,
132.16, 130.82, 130.74, 127.99, 127.72, 126.94, 126.74, 126.46,
124.57, 121.16, 120.33, 116.72, 116.33, 116.11, 115.75, 107.11, 71.97;
HRMS of [C31H21N8O5Br + H]+ (m/z) 665.1849; Calcd: 665.1840.
2.1.5.8. N-((1-(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-
yl)methylene)−4-((1-(4–methoxy
phenyl)−1H-1,2,3-triazol-4-
yl)methoxy)aniline (9 h). FT-IR νmax (cm−1): 3051 (=C H), 2838
(C–H), 1605 (C = C), 1531 (C = N); 1332 (NO2); 1H NMR (400 MHz,
DMSO–d6) δ 10.00 (s, 1H, CH-triazole), 9.49 (s, 1H, CH), 9.38 –
9.29 (m, 1H, Ar-H), 8.89 (d, J = 2.5 Hz, 1H, Ar-H), 8.87 – 8.82 (m,
1H, Ar-H), 8.66 (dd, J = 9.0, 2.5 Hz, 1H, Ar-H), 8.36 (d, J = 8.9 Hz,
2H, Ar-H), 8.18 (d, J = 3.2 Hz, 1H, Ar-H), 7.88 (d, J = 8.7 Hz, 1H,
Ar-H), 7.75 (dd, J = 18.5, 8.7 Hz, 1H, Ar-H), 7.47 (d, J = 8.6 Hz,
2H, Ar-H), 7.35 (s, 1H, CH), 7.18 – 6.95 (m, 5H, Ar-H), 5.18 (s,
2H, CH2), 3.83 (s, 3H, OCH3); 13C NMR (100 MHz, DMSO–d6)
δ 168.92, 162.65, 160.29, 154.91, 145.91, 142.70, 138.99, 138.89,
135.11, 131.83, 131.01, 129.90, 127.87, 126.61, 126.05, 123.78, 122.44,
121.12, 120.65, 116.62, 114.58, 114.06, 113.98, 106.83, 71.97, 55.29;
HRMS of [C32H24N8O6 + H]+ (m/z) 617.1768; Calcd: 617.1789.
–
2.1.5.4. N-((1-(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-
yl)methylene)−4-((1-(4-nitro
phenyl)−1H-1,2,3-triazol-4-
yl)methoxy)aniline (9d). FT-IR νmax (cm−1): 3065 (=C H), 2931
(C–H), 1593 (C = C), 1552 (C = N); 1323 (NO2); 1H NMR (400 MHz,
DMSO–d6) δ 9.73 (s, 1H, CH-triazole), 9.49 (s, 1H, CH), 8.85 (d,
J = 2.3 Hz, 1H, Ar-H), 8.72 – 8.49 (m, 5H, Ar-H), 8.38 (s, 1H, CH),
8.16 (d, J = 9.0 Hz, 1H, Ar-H), 7.63 (d, J = 8.5 Hz, 2H, Ar-H), 7.36
(d, J = 3.5 Hz, 2H, Ar-H), 7.06 (d, J = 7.5 Hz, 1H, Ar-H), 6.94 (d,
J = 8.4 Hz, 2H, Ar-H), 6.84 (d, J = 8.5 Hz, 2H, Ar-H), 5.12 (s, 2H,
CH2); 13C NMR (100 MHz, DMSO–d6) δ 158.25, 154.58, 144.60,
141.90, 135.62, 132.18, 131.86, 130.01, 129.27, 127.91, 127.65, 127.18,
124.10, 123.57, 122.43, 121.31, 121.12, 116.57, 116.23, 115.90, 115.77,
115.59, 115.35, 106.70, 71.96; HRMS of [C31H21N9O7 + H]+ (m/z)
632.2234; Calcd: 632.2230.
–
2.1.5.9. N-((1-(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-
2.1.5.5. N-((1-(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-
yl)methylene)−4-((1-(p-tolyl)−1H-1,2,3-triazol-4-yl)methoxy)aniline
yl)methylene)−4-((1-(3–methoxy
phenyl)−1H-1,2,3-triazol-4-
(9i). FT-IR νmax (cm−1): 3065 (=C H), 2839 (C–H), 1602 (C = C),
–
yl)methoxy)aniline (9e). FT-IR νmax (cm−1): 3065 (=C H), 2961
(C–H), 1582 (C = C), 1532 (C = N); 1323 (NO2); 1H NMR (400 MHz,
DMSO–d6) δ 9.49 (s, 1H, CH-triazole), 9.36 (s, 1H, CH), 8.89 (d,
J = 5.7 Hz, 1H, Ar-H), 8.66 (d, J = 8.6 Hz, 1H, Ar-H), 8.37 (d,
J = 8.9 Hz, 1H, Ar-H), 8.30 (d, J = 8.9 Hz, 1H, Ar-H), 8.18 (t,
J = 8.8 Hz, 1H, Ar-H), 7.48 (t, J = 7.8 Hz, 1H, Ar-H), 7.43 – 7.30
(m, 5H, Ar-H), 7.23 (d, J = 8.7 Hz, 2H, Ar-H), 7.10 (d, J = 7.8 Hz,
1H, Ar-H), 7.05 (s, 1H, CH), 6.96 (d, J = 7.1 Hz, 2H, Ar-H), 5.20
(s, 2H, CH2), 3.81 (s, 3H, OCH3); 13C NMR (100 MHz, DMSO–d6)
δ 168.88, 159.50, 154.77, 146.03, 142.76, 135.07, 131.92, 131.45,
130.29, 130.06, 129.79, 127.89, 126.75, 126.28, 122.96, 121.20,
121.12, 121.07, 120.81, 120.77, 120.41, 116.77, 114.89, 114.11, 111.20,
107.33, 71.97, 55.19; HRMS of [C32H24N8O6 + H]+ (m/z) 617.2476;
Calcd: 617.2475.
–
1531 (C = N); 1336 (NO2); 1H NMR (400 MHz, DMSO–d6) δ 8.89 (s,
1H, CH-triazole), 8.28 (d, J = 7.5 Hz, 2H, Ar-H), 8.11 (d, J = 7.1 Hz,
2H, Ar-H), 7.98 (s, 1H, CH), 7.88 (d, J = 8.7 Hz, 2H, Ar-H), 7.82 (dd,
J = 7.7, 6.6 Hz, 1H, Ar-H), 7.78 – 7.69 (m, 4H, Ar-H), 7.69 – 7.62 (m,
2H, Ar-H), 7.46 (t, J = 7.4 Hz, 1H, Ar-H), 7.27 (s, 1H, CH), 7.14 (d,
J = 8.7 Hz, 2H, Ar-H), 5.41 (s, 2H, CH2), 2.17 (s, 3H, CH3); 13C NMR
(100 MHz, DMSO–d6) δ 166.24, 161.59, 158.76, 154.09, 146.55,
144.46, 144.18, 134.23, 133.93, 131.02, 130.85, 130.16, 128.10,
127.25, 127.21, 126.89, 126.34, 126.14, 126.05, 125.81, 125.69,
124.59, 121.53, 115.27, 61.99, 22.84; HRMS of [C32H24N8O5 + H]+
(m/z) 601.9398; Calcd: 601.9388.
2.1.5.10. N-((1-(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-
yl)methylene)−4-((1-(3-nitro
phenyl)−1H-1,2,3-triazol-4-
yl)methoxy)aniline (9j). FT-IR νmax (cm−1): 3003 (=C H), 2838
–
2.1.5.6. N-((1-(2,4-dinitrophenyl)−3-phenyl-1H-pyrazol-4-
yl)methylene)−4-((1-(2–methoxy
phenyl)−1H-1,2,3-triazol-4-
(C–H), 16,014 (C
=
C), 1532 (C
=
N); 1337 (NO2); 1H NMR
yl)methoxy)aniline (9f). FT-IR νmax (cm−1): 3065 (=C H), 2839
(C–H), 1602 (C = C), 1531 (C = N); 1313 (NO2); 1H NMR (400 MHz,
DMSO–d6) δ 9.54 (s, 1H, CH-triazole), 9.36 (s, 1H, CH), 8.89 (d,
J = 2.3 Hz, 1H, Ar-H), 8.85 (s, 1H, Ar-H), 8.67 (dd, J = 8.9, 2.3 Hz,
1H, Ar-H), 8.39 (d, J = 8.9 Hz, 1H, Ar-H), 8.19 (d, J = 9.0 Hz, 2H,
Ar-H), 8.15 (d, J = 9.5 Hz, 2H, Ar-H), 7.87 (d, J = 2.5 Hz, 1H, Ar-H),
7.54 (t, J = 7.3 Hz, 1H, Ar-H), 7.42 (d, J = 7.4 Hz, 1H, Ar-H), 7.30 –
7.20 (m, 2H, Ar-H), 7.16 – 7.00 (m, 3H, Ar-H), 6.96 (s, 1H, CH), 5.21
(s, 2H, CH2), 3.78 (s, 3H, OCH3); 13C NMR (100 MHz, DMSO–d6)
δ 168.96, 156.65, 156.22, 153.68, 145.94, 142.76, 141.70, 140.59,
135.20, 131.87, 131.56, 131.31, 129.21, 128.80, 128.52, 127.92, 126.68,
(400 MHz, DMSO–d6) δ 8.85 (s, 1H, CH-triazole), 8.70 (s, 1H, CH),
8.28 (dd, J = 9.7, 3.7 Hz, 1H, Ar-H), 7.84 (dd, J = 8.7, 4.3 Hz, 2H,
Ar-H), 7.77 – 7.70 (m, 1H, Ar-H), 7.66 (dd, J = 7.8, 2.2 Hz, 1H,
Ar-H), 7.59 (dd, J = 9.0, 7.3 Hz, 2H, Ar-H), 7.49 – 7.41 (m, 5H,
Ar-H), 7.26 (s, 1H, CH), 7.12 (t, J = 7.5 Hz, 1H, Ar-H), 6.98 (dd,
J = 11.6, 8.8 Hz, 2H, Ar-H), 6.91 (d, J = 7.9 Hz, 1H, Ar-H), 5.22 (s,
2H, CH2); 13C NMR (100 MHz, DMSO–d6) δ 166.30, 161.59, 158.75,
158.53, 146.53, 137.83, 136.88, 134.90, 134.23, 130.75, 129.43,
128.94, 128.75, 127.21, 126.86, 126.80, 126.76, 126.34, 125.83,
125.44, 123.49, 121.54, 119.77, 117.44, 115.12, 114.99, 67.26; HRMS
of [C31H21N9O7 + Na]+ (m/z) 654.2394; Calcd: 654.2400.
–
3

K. Suryanarayana, A.R. Robert, N. Kerru et al.
Journal of Molecular Structure 1243 (2021) 130865
2.2. Anticancer studies for in vitro screening
only 5 amino acid residues with five hydrogen bonds, 2 electro-
static bonds, and 2 hydrophobic bonds of the breast adenocarci-
noma protein (PDB ID: 6sbo), while 9a found to interact with 11
amino acid residues with 4 hydrogen bonds, 3 electrostatic bonds
and 7 hydrophobic bonds, 9c found to interact with 9 amino acid
residues with 2 hydrogen bonds, 4 electrostatic bonds, 3 hydropho-
bic bonds and 1 other (pi-sulfur) bond, 9e found to interact with
13 amino acid residues with 4 hydrogen bonds, 1 electrostatic
bond and 5 hydrophobic bonds, and 9i found to interact with 14
amino acid residues with 3 hydrogen bonds, 1 electrostatic bond,
11 hydrophobic bonds and 1 other (pi-sulfur) bond. This may be
the reason for the enhanced activity of the compounds. Similarly,
Combretastatin-A4 is found to interact with only 6 amino acid
residues with 3 hydrogen bonds, and 4 hydrophobic bonds of the
cervical carcinoma protein (PDB ID: 2q79), but 9f found to inter-
act with 7 amino acid residues with 6 hydrogen bonds, 1 electro-
static bond, 4 hydrophobic bonds and one other (pi-sulfur) bond,
9i found to interact with 5 amino acid residues with 3 hydro-
gen bonds, 4 hydrophobic bonds and one other (pi-sulfur) bond,
and 9j found to interact with 9 amino acid residues with 7 hy-
drogen bonds and 8 hydrophobic bonds. Likewise, Combretastatin-
A4 is found to interact with only 6 amino acid residues with 4
hydrogen bonds, 2 hydrophobic bonds and 2 other (pi-sulfur) of
the human epithelial colorectal adenocarcinoma protein (PDB ID:
2maw), but 9a found to interact with 8 amino acid residues with
2 hydrogen bonds, and 7 hydrophobic bonds, 9c found to interact
with 7 amino acid residues with 3 hydrogen bonds, 6 hydrophobic
bonds and one other (pi-sulfur) bond, 9i found to interact with 8
amino acid residues with 2 hydrogen bonds, 7 hydrophobic bonds
and one other (pi-sulfur) bond, and 9j found to interact with 7
amino acid residues with 4 hydrogen bonds and 9 hydrophobic
bonds. Figs. S1 to S33 of supplementary information give the 3D
and 2D representation of the docked molecules with the respec-
tive proteins.
2.2.1. Cell culture
The breast adenocarcinoma (MCF-7), cervical carcinoma (HeLa),
and human epithelial colorectal adenocarcinoma (Caco-2) cell lines
were employed for this investigation. MCF-7, HeLa, and Caco-
2 cell lines were developed in EMEM supplemented with 10%
γ -irradiated foetal bovine serum (FBS), penicillin (100 U mL−1),
streptomycin (100 μM), at 37 °C with 5% CO2 in a moistened atmo-
sphere. Cells were subjected to split at ratios (1:3–1:5) every 3–4
days at semi to full convergence. All the cells were newly cultured
and plated 24 h earlier to each test to permit cells’ attachment.
The complete media preparation, cell lines propagation, mainte-
nance, and cell culture process were carried under the class-II Air
volution biological safety cabinet (United Scientific Pvt Ltd). All the
chemical details are provided in supporting information (SI-2).
2.2.2. Procedure
Cytotoxicity evaluation of the synthetic molecules against all
cell lines was measured using MTT reduction process via in
vitro method [45]. MCF-7, HeLa, and Caco-2 cells were dis-
cretely trypsinized and plated in 96-multiwell plates (density 2.1–
2.5 × 104 cells/well) and left overnight at 37° for incubation in
0.1 mL complete growth medium to attain 80% confluence. Next,
the cells were exposed to different concentrations (25 to 200 μM
in DMSO) of target molecules, followed by another 48 h incubation.
After incubation is complete, the medium was removed from the
wells, 0.1 mL of fresh medium, MTT reagent (5 mg/mL in sterile
PBS at pH 7.4) was added and incubated for 4 h. The medium was
removed again, and wells were replaced with DMSO (0.1 mL). Ab-
sorbance readings were noted at 570 nm with a Vacutec MR-96A
microplate reader. All the results were equated to the cytotoxicity
properties of Combretastatin-A4 as a standard drug [46,47].
Table S1 indicates that the unfavorable non-bonds (bump) of
breast adenocarcinoma protein (PDB ID: 6sbo) with 9b, 9c, 9f and
9i, cervical carcinoma protein (PDB ID: 2q79) with 9b, 9c, 9f and
9i, and human epithelial colorectal adenocarcinoma protein (PDB
ID: 2maw) with 9c and Combretastatin-A4. All the molecules’ own
unsatisfied sites are mostly electron-rich or poor sites with all the
proteins. Hydrogen atoms act as always donors, oxygen act as ac-
ceptor and nitrogen act as both donor and acceptor during bind.
2.3. Docking analysis
The molecules are optimized to the minimum geometry us-
ing B3LYP/6–311+G (2d,p) level of theory and this obtained ge-
ometry has been used for docking studies. For docking, proteins
responsible for the various cancer studies has been downloaded
from the RSC protein data bank [48,49]. The docking has been per-
formed using Autodock 4.0 [50] and visualisations using Biodiscov-
ery Studio. Breast adenocarcinoma protein (PDB ID: 6sbo) [51], cer-
vical carcinoma protein (PDB ID: 2q79) [52], and human epithelial
colorectal adenocarcinoma membrane receptor protein (PDB ID:
2maw) [53] are downloaded and prepared before effective dock-
ing. Proteins downloaded from the RCSB site, contain ligand and
water molecules that are cleared. Then polar hydrogen atoms were
added, noted x, y, z values from attributes of the SBD site sphere,
and finally this protein file was saved for the next step to docking
work. The grid box values are set at 30, 30, and 30 for x, y, and
z directions, again delete water molecules, added polar hydrogen
atoms, and added Kollman charges with no other charges. Then
input ligand (optimized) molecule to prepare ligand for docking.
Docking was done in a setting of 300 iterations.
3. Results and discussion
3.1. Chemistry
The target molecules’ synthesis is depicted in Scheme 1. 1-
(2,4-dinitrophenyl)−2-(1-phenylethylidene)-hydrazine (3) was syn-
thesized via condensation of acetophenone with 2,4-dinitrophenyl
hydrazine. Compound (3) on further reaction with a solution of
DMF and POCl3 via cyclization affords pyrazole-4-carbaldehyde (4).
Later, compound (4) on condensation with para-amino phenol un-
der reflux condition yielded pyrazol-4-yl-methyleneaminophenol
(6). Compound (6) was O-alkylated by propargyl bromide in the
presence of a strong base under mild reaction conditions, affording
the corresponding compound (7) in good yields. Further, molecule
(7) on cyclization with various substituted azides produced the de-
sired pyrazole bearing triazole derivatives (9a–j).
Table 3 shows the type of amino acid residues in the respective
proteins interacting with the compounds and the standard drug.
Table S1 (Supporting information) presents the nature of non-
bonding (favourable, unfavorable, and unsatisfied) interactions like
conventional hydrogen bonds, carbon-hydrogen bonds, pi-cation,
pi-anion, alkyl-alkyl, and pi-alkyl, pi-sigma, pi-pi T-shaped, amide-
pi stacked, pi-sulfur, pi-pi stacked, pi-donor hydrogen bonds, and
unfavorable bump (steric). We try to explain the biological activi-
ties of the compounds based on these interactions. Favourable non-
bonds from the tables show that Combretastatin-A4 interacts with
These novel molecules were validated and characterized em-
ploying 1H NMR, 13C NMR, FT-IR and HR-MS analysis. The pro-
ton NMR (1H) spectrum of the molecule 9 h exhibited three sin-
glet peaks at δ 10.00, 9.49 and 7.35 ppm confirming the pres-
ence of CH-triazole, CH-imine and CH-pyrazole groups. The pres-
ence of multiplet peaks at δ 6.95–8.89 ppm approves the presence
of aromatic protons. Two singlet peaks were detected at δ 5.18 and
4

K. Suryanarayana, A.R. Robert, N. Kerru et al.
Journal of Molecular Structure 1243 (2021) 130865
Scheme 1. Synthesis of novel dinitrophenylpyrazole bearing 1,2,3-triazole derivatives.
3.83 ppm which are attributed to CH2 and methoxy groups. Fur-
ther, the structure of the target molecule was confirmed by their
13C NMR data. Moreover, the molecular ion peak was found at m/z
617.1768 and calculated value at 617.1789 (M + H)+ in the high-
resolution mass spectrum, which is in good agreement with the
molecular formula C32H24N8O6 of compound 9 h.
ues are in the range of 4.0–16.0 μM, which manifests their po-
tential anti-cancer activity. The positive control Combretastatin-A4
exhibited the values between (IC50 = 9.0 and 11.0 μM against
screened cell lines. The molecules 9e and f showed the extreme
growth inhibitory activity against the cervical carcinoma (HeLa)
cell line (IC50 = 4.0 μM and 5.0 μM) higher than that of the refer-
ence drug. The molecule 9 h was found to be more potent against
breast adenocarcinoma (MCF-7) cell line (IC50 value 8.0 μM). Fur-
ther, compounds 9a–c exhibited potent cytotoxic activity against
the human epithelial colorectal adenocarcinoma (Caco-2) cell line.
Structure-activity relationship (SAR) investigations reveal that in
compounds 9e and 9f, the methoxy substitution on the phenyl
triazole moiety tagged with pyrazole leads to higher potential ac-
tivity than the reference drug Combretastatin-A4 against the Hela
cell lines. However, the molecule 9 h also exhibited potential cy-
totoxic activity against HeLa and MCF-7 cell lines. The above re-
sults indicated that the molecules 9e, 9f, and 9 h with methoxy
group on phenyl ring at the ortho, meta, or para position of tar-
get molecule possess potent activity. Molecules 9a, 9b, and 9c with
halo functional groups (at ortho and para) have displayed good ac-
tivity against Caco-2 cell line. Therefore, electron-donating groups
like methoxy on the phenyl ring enriched the compounds’ activ-
ity, while electron-withdrawing functional groups such as Cl and
Br retarded the cytotoxic activity. Hence, it can be attributed that
electron-donating groups in the triazole core help improve these
compounds’ anti-cancer activity.
3.2. Biological screening
The in vitro screening of the synthesized molecules (9a–j) was
done against three cancer cell lines, namely breast adenocar-
cinoma (MCF-7), cervical carcinoma (HeLa), and human epithe-
lial colorectal adenocarcinoma (Caco-2) using the MTT assay ap-
proach. The IC50 values of the synthesized molecules compared to
Combretastatin-A4 as a reference drug and DMSO as a negative
control are provided in Table 1. Most of the molecules’ IC50 val-
Table 1
Cytotoxic activity of novel dinitrophenylpyrazole bearing 1,2,3-triazole
scaffolds (9a–j) against human cancer cell lines.
IC50 (mean SD) μMa
Entry Product
MCF-7
HeLa
Caco2
1
9a
14
12
11
11
10
11
15
08
12
12
10
0.22
12
15
10
16
04
05
12
06
11
10
0.44
0.58
0.29
0.35
0.47
0.05
0.11
0.38
0.10
0.21
10
09
10
14
12
15
14
12
15
12
11
0.11
0.23
0.47
0.33
0.19
0.02
0.09
0.21
0.31
0.54
0.35
2
9b
0.15
0.29
0.17
0.57
0.10
0.42
0.19
021
3
9c
4
9d
5
9e
6
9f
3.3. Molecular docking screening
7
9 g
8
9h
Table 2 shows the binding affinity of molecules 9a-9j against
breast adenocarcinoma protein (PDB ID: 6sbo), cervical carcinoma
protein (PDB ID: 2q79), and human epithelial colorectal adenocar-
cinoma protein (PDB ID: 2maw) compared to the standard drug
Combretastatin-A4 using blind-drug docking. All the molecules
9
9i
10
11
9j
0.37
0.40
Combretastatin-A4
9
0.32
a
IC50 value corresponds to the concentration of the respective com-
pound required to effect 50% mortality in net cells.
5

K. Suryanarayana, A.R. Robert, N. Kerru et al.
Journal of Molecular Structure 1243 (2021) 130865
Table 2
around −10 kcal/mol. Combretastatin-A4 showed binding energy
of −5.50 kcal/mol with the cervical carcinoma protein (PDB ID:
2q79B). All the test molecules showed higher affinity than this
molecule and the maximum affinity was showed by 9i, 9f and 9j
with values around −7.50 kcal/mol. The same trend is observed
for human epithelial colorectal adenocarcinoma protein (PDB ID:
2maw) with a binding affinity of −7.50 kcal/mol for the stan-
dard and all molecules showed more affinity than that. All the test
molecules showed higher affinity than this molecule and the max-
imum affinity was showed by 9a, 9c, 9i and 9j with values around
−9.50 kcal/mol.
Binding affinity for molecule 9a–j and standard drug
Combretastatin-A4 against breast adenocarcinoma pro-
tein (PDB ID: 6sbo), cervical carcinoma protein (PDB ID:
2q79), and human epithelial colorectal adenocarcinoma
protein (PDB ID: 2maw).
Binding affinity (in kcal/mol)
Compounds
6sbo
2q79
2maw
9a
−10.1
−9.8
−10.0
−9.8
−10.0
−9.8
−8.1
−7.4
−10.1
−8.4
−6.6
−7.5
−6.9
−6.8
−6.8
−7.6
−7.7
−7.7
−6.6
−7.9
−7.7
−5.5
−10.1
−7.7
−9.5
−8.9
−8.5
−8.2
−8.5
−8.5
−9.8
−9.5
−7.5
9b
9c
9d
Figs. 1–3 give the 3D representation of the interaction between
the drug molecule 9a and the target proteins. The figures repre-
senting 2D interactions are provided in the Supplementary Infor-
mation (Figs. SI–S3).
9e
9f
9 g
9h
9i
The compounds showed better docking when compared to that
of the standard drug Combretastatin-A4, which is widely used
against these cancers. It is found that there are standard drug in-
teracts with GLU A:385, ASN A:455, TYR A:459, LEU A:511, and
ARG A:515 amino acid residues (5 nos) of the PDB:6sbo. The drug
with maximum efficiency is 9a and it interacts with the same pro-
tein as follows: GLU A:323, PRO A:324, ILE A:326, GLU A:353, MET
A:357, ILE A:386, LEU A:387, TRP A:393, ARG A:394, HIS A:398, and
LYS A:449 (11 nos). The interaction between 9a and the protein
residues is more than twice that of the standard drug, which is
the reason for enhanced activity.
9j
Combretastatin-A4
showed more significant activity and binding towards the three
cancer proteins under study in agreement with experimental re-
sults. The standard drug’s estimated binding affinity towards ade-
nocarcinoma protein (PDB ID: 6sbo) was −6.60 kcal/mol. All the
test molecules showed higher affinity than this molecule and the
maximum affinity was showed by 9a, 9c, 9e and 9i with values
Table 3
Label for breast adenocarcinoma protein (PDB ID: 6sbo), cervical carcinoma protein (PDB ID: 2q79), and human epithelial colorectal adenocarcinoma protein (PDB ID:
2maw) residues interacting with the molecules and the standard drug.
Compounds
Protein residues
Molecules 9a-j against breast adenocarcinoma protein 6sbo residues
9a
9b
GLU A:323, PRO A:324, ILE A:326, GLU A:353, MET A:357, ILE A:386, LEU A:387, TRP A:393, ARG A:394, HIS A:398, and LYS A:449
LEU A:346, THR A:347, ALA A:350, ASP A:351, GLU A:353, LEU A:387, LEU A:391, ARG A:394, PHE A:404, MET A:421, ILE A:424, LEU
A:525, VAL A:534, PRO A:535, and LEU A:539
9c
9d
9e
GLUA:323, PRO A:324, ILE A:326, TRP A:393, ARG A:394, MET A:396, LEU A:397, and LYS A:449
GLU A:323, PRO A:324, PRO A:325, ILE A:326, GLY A:390, TRP A:393, ARG A:394, and LYS A:449
LEU A:346, THR A:347, ALA A:350, ASP A:351, LEU A:387, PHE A:404, MET A:421, ILE A:424, LEU A:525, ASN A:532, PRO A:535, SER
A:536, and LEU A:539
9f
LEU A:346, THR A:347, ALA A:350, ASP A:351, GLU A:353, LEU A:387, ARG A:394, PHE A:404, MET A:421, ILE A:424, LEU A:425, ASN
A:532, PRO A:535, and LEU A:539
9 g
9h
9i
GLU A:323, PRO A:324, GLU A:353, TRP A:393, ARG A:394, GLN A:441, and LYS A:449
GLU A:323, PRO A:324, ILE A:326, TRP A:393, ARG A:394, LEU A:403, GLN A:441, GLY A:442, and LYS A:449
MET A:343, LEU A:346, THR A:347, ALA A:350, ASP A:351, GLU A:353, LEU A:387, ARG A:394, PHE A:404, MET A:421, ILE A:424, LEU
A:525, PRO A:535, and LEU A:539
9j
GLU A:323, PRO A:324, PRO A:325, ILE A:326, GLY A:390, TRP A:393, ARG A:394, and LYS A:449
GLU A:385, ASN A:455, TYR A:459, LEU A:511, and ARG A:515
Combretastatin-A4
Molecules 9a-j with cervical carcinoma protein 2q79 residues
9a
ALA A:312, SER A:314, SER A:315, TRP A:317, and MET A:361
9b
ARG A:302, ILE A:350, PRO A:351, LYS A:352, and VAL A:356
9c
ARG A:302, ILE A:350, PRO A:351, LYS A:352, and VAL A:356
9d
ARG A:302, ILE A:350, PRO A:351, LYS A:352, THR A:355, and VAL A:356
ARG A:302, LYS A:349, ILE A:350, PRO A:351, LYS A:352, ILE A:354, and VAL A:356
HIS A:288, SER A:314, SER A:315, TRP A:317, THR A:332, THR A:334, and MET A:361
HIS A:288, ALA A:312, SER A:315, TRP A:317, and MET A:361
9e
9f
9 g
9h
ALA A:312, SER A:314, SER A:315, TRP A:317, and MET A:361
9i
HIS A:288, ALA A:312, TRP A:317, THR A:334, MET A:361
9j
ILE A:286, HIS A:288, THR A:311, ALA A:312, VAL A:313, SER A:314, SER A:315, TRP A:317, and MET A:361
ILE A:286, HIS A:288, ALA A:312, THR A:332, THR A:334, and MET A:361
Combretastatin-A4
Molecules 9a-j with human epithelial colorectal adenocarcinoma protein 2maw residues
9a
TYR A:72, SER A:75, ILE A:78, ILE A:79, VAL A:118, and PHE A:119
9b
ASN A:12, PRO A:16, LEU A:19, LEU A:46, SER A:47, VAL A:50, PHE A:51, and LEU A:54
VAL A:18, ALA A:22, LEU A:25, ARG A:110, SER A:117, VAL A:118, and ILE A:121
LEU A:7, GLY A:10, LEU A:14, VAL A:18, THR A:76, SER A:117, THR A:120, ILE A:121, ILE A:127, and LEU A:128
VAL A:18, ALA A:22, LEU A:25, ARG A:110, MET A:114, SER A:117, VAL A:118, and ILE A:121
TYR A:72, SER A:75, ALA A:115, VAL A:118, ILE A:122, GLY A:126, and MET A:129
GLY A:10, LEU A:11, LEU A:14, VAL A:18, ILE A:69, THR A:120, ILE A:121, THR A:124, and ILE A:127
GLY A:10, LEU A:11, LEU A:14, CYS A:17, VAL A:18, SER A:117, THR A:120, ILE A:121, and THR A:124
TYR A:72, SER A:75, ILE A:79, VAL A:118, PHE A:119, ILE A:122, GLY A:126, and MET A:129
VAL A:18, LEU A:19, ALA A:22, LEU A:23, LEU A:25, SER A:117, THR A:120
9c
9d
9e
9f
9 g
9h
9i
9j
Combretastatin-A4
LEU A:13, CYS A:17, PHE A:73, THR A:76, CYS A:123, and THR A:124
6

K. Suryanarayana, A.R. Robert, N. Kerru et al.
Journal of Molecular Structure 1243 (2021) 130865
4. Conclusion
In conclusion, herein we report the synthesis of a novel dini-
trophenylpyrazole bearing 1,2,3-triazole derivatives (9a–j). All the
obtained molecules were evaluated and characterized for their in-
fluence on cellular feasibility against three cancer cell lines MCF-7,
HeLa, and Caco-2, via the MTT approach. Amongst all the screened
molecules, 9e, 9f, and 9 h were found to have the potent growth
inhibitory activity against the HeLa cell line (IC50 = 4.0 μM,
5.0 μM and 6.0 μM). Besides, compound 9 h was found most po-
tent against MCF-7 cell line IC50 value of 8.0 μM. This investigation
concludes that further structural improvement is a prerequisite for
developing a novel therapeutic agent. Molecular docking studies
indicate that all the test molecules show considerably high activity
against breast adenocarcinoma, cervical carcinoma, and human ep-
ithelial colorectal adenocarcinoma cancer proteins. Hence, all these
molecules seem to be ideal candidates to be further investigated
as preferred anti-cancer drugs.
Declaration of Competing Interest
Fig. 1. Skeletal structure of interactions between 9a and breast adenocarcinoma
protein.
The authors have declared no conflict of interest
CRediT authorship contribution statement
Kotyada Suryanarayana: Investigation, Conceptualization.
Alice Rinky Robert: Writing - review & editing, Conceptualiza-
tion. Nagaraju Kerru: Investigation, Writing - review & editing.
Thangaiyan Pooventhiran: Writing
- review & editing, Inves-
tigation. Renjith Thomas: Resources, Formal analysis. Suresh
Maddila: Conceptualization, Supervision. Sreekantha B. Jonnala-
gadda: Writing - review & editing.
Acknowledgments
The authors are very grateful to the University of KwaZulu
Natal, Durban, South Africa, and GITAM Deemed to be Univer-
sity, Bengaluru and Visakhapatnam, India for financial and research
support.
Supplementary materials
Supplementary material associated with this article can be
found, in the online version, at doi:10.1016/j.molstruc.2021.130865.
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