Design, synthesis, molecular docking, admet studies and biological evaluation of pyrazoline incorporated 1, 2, 3-triazole benzene sulphonamides

Article abstract of DOI:10.22159/ijpps.2019v11i6.32684

Objective: The main objective of this work was to design, synthesize and evaluate the novel pyrazoline incorporated 1,2,3-triazole benzene sulphonamides for cytotoxic and anti-gout activities also to perform Insilco molecular docking studies. Methods: Des

Full text of DOI:10.22159/ijpps.2019v11i6.32684

International Journal of Pharmacy and Pharmaceutical Sciences
Print ISSN: 2656-0097 | Online ISSN: 0975-1491
Vol 11, Issue 6, 2019
Original Article
DESIGN, SYNTHESIS, MOLECULAR DOCKING, ADMET STUDIES AND BIOLOGICAL
EVALUATION OF PYRAZOLINE INCORPORATED 1, 2, 3-TRIAZOLE BENZENE
SULPHONAMIDES
SRAVANTHI SILIVERI1,2*, NAGARAJU BASHABOINA1, HARINADHA BABU VAMARAJU1, SHIVA RAJ3
1Department of Pharmaceutical Chemistry, G. Pulla Reddy College of Pharmacy, Mehdipatnam, Hyderabad, Telangana, 500028, India,
2Department of Pharmacy, OUCT, Osmania University, Hyderabad, 500007, India, 3Department of Chemistry, Osmania University,
Hyderabad, 500007, India
Email: sravanthisiliveri@gmail.com
Received: 19 Feb 2019 Revised and Accepted: 16 Apr 2019
ABSTRACT
Objective: The main objective of this work was to design, synthesize and evaluate the novel pyrazoline incorporated 1,2,3-triazole benzene
sulphonamides for cytotoxic and anti-gout activities also to perform Insilco molecular docking studies.
Methods: Designed compounds were synthesized by condensation of different substituted chalcones (3a-i) with hydrazine hydrate and substituted
phenylhydrazines. All the synthesized compounds were characterized on the basis of physical and spectral data. To predict the affinity and activity
of the ligand molecule Libdock program was employed to generate different bioactive binding poses of designing molecules at the active site of
protein Phosphatidylinositol 3-kinase (PI3Kα). Title compounds were evaluated for cytotoxic activity by using 3-(4, 5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide (MTT) assay and anti-gout activity by potassium oxonate induced assay.
Results: All the synthesized compounds showed characteristic peaks in FTIR, 1H, 13C NMR and MASS spectral analysis. In molecular docking studies,
compound 3i has shown good binding affinity to the active site of PI3Kα with a docking score of 145.031 and 4 hydrogen bonding interactions with
least hepatotoxicity and good bioavailability when compared with that of reference ligand KKR exhibited a Libdock score of 88.35. Remaining
compounds also have a good binding affinity with a minimum of 2 bonding interactions and having better absorption, distribution, metabolism,
elimination and toxicity (ADMET) profile. The same compound (3i) exhibited the highest cytotoxic activity with an IC50 value of 4.54µg/ml.
Compound 4d was evaluated for anti-inflammatory activity and it has significantly ameliorated against potassium oxonate induced gout in mice
when compared with that of standard drug allopurinol due to its anti-inflammatory property.
Conclusion: We designed and synthesized a novel series of title compounds in quantitative yields and performed docking studies. New derivatives
have a good binding affinity towards PI3Kα enzyme, good bioavailability, least hepatotoxicity and significant cytotoxic activity.
Keywords: ADMET, Cancer, Docking, Malignant, PI3Kα, Proliferation
© 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open-access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijpps.2019v11i6.32684
INTRODUCTION
drug discovery and therefore pyrazole ring constitutes a relevant
synthetic target in the pharmaceutical industry. On the other hand,
1,2,3-triazole is an important heterocycle which has gained a lot of
interest for researchers in view of its high potency, low toxicity with
broad spectrum of activities. Triazole derivatives have been
reported to have anticancer [22], anti-inflammatory [23],
antibacterial [24], antiviral [25], anti-human immuno virus (HIV)
[26], fungicidal [27] and insecticidal [28-30] activities. The objective
of the current work was to synthesize novel pyrazoline
incorporating 1,2,3-triazole benzene sulphonamide derivatives as
potent antiproliferative and anti-inflammatory agents. We were well
known about the synthesis of simple pyrazolines from chalcones but
we designed pyrazoline incorporating 1,2,3-triazolyl benzene
sulphonamide derivatives. Inspired by the biological properties of
both pyrazole and triazole heterocycles in the present study, we
thought of synchronizing both these moieties into a single molecule
in order to obtain new hybrid molecules with improved biological
activity and low toxicity.
Cancer is a multifactorial disease, arising from the uncontrolled
proliferation of a cell with the potential to invade to other organs of
the body [1]. PI3Kα is one of the proto-oncogenes, which has a vital
role in the regulation of many important cell signaling pathways
including cellular replication, cell proliferation leading to growth
and apoptosis [2, 3]. The enzyme PI3Kα appeared to involve in 40%
of all types of human cancers including breast, gastric, cervical,
urinary tract, colon, non-small-cell lung and squamous cell lung
carcinomas [4]. Considerable evidence suggest that PI3K (P110α)
subunit protein is expected to mutate in these cancers that bring
about constitutive downstream pathways leading to defective
control of cellular proliferation and malignant transformation, thus
PI3Kα is drawing attention in cancer biology [5,6]. Hence targeting
PI3Kα may be considered as a promising approach in the design of
new anticancer drugs.
Pyrazoles are a class of heterocyclic compounds characterized by 5-
membered aromatic ring structure composed of three carbon atoms
and two nitrogen atoms in adjacent positions. Pyrazole derivatives
have a long history of applications in agriculture as herbicides and
insecticides as well as in the pharmaceutical industry as antipyretic
and anti-inflammatory agents [7-12]. Pyrazole derivatives have been
reported to show a broad spectrum of biological activity including
antibacterial [13], antifungal [14], analgesic [15], anti-inflammatory
[16-18], neuroprotective [19], estrogen receptor binding [20],
antineoplastic [21], activities. Due to their wide range of biological
activities, pyrazoles received considerable interest in the field of
MATERIALS AND METHODS
All the chemicals and solvents used were of synthetic grade from SD
fine chemicals Ltd., (Mumbai, India), and avra chemicals pvt ltd
Hyderabad. Completion of the reaction was monitored by analytical
thin layer chromatography (TLC) using E-Merck 0.25 mm silica gel
plates. Visualization was accomplished with ultraviolet (UV) light
(256 nm) and iodine chamber. Synthesized compounds were
purified by the re-crystallization process. The purity of the
compounds was checked by a single spot in TLC and solvent system

Siliveri et al.
Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
for TLC was determined on trial and error basis. Melting points were
determined in open capillary tubes using ANALAB melting point
apparatus and were uncorrected. All the 1H NMR spectra were
recorded on varian 400 MHz spectrometer using DMSO-d6 as solvent
and tetramethylsilane (TMS) as an internal standard. Chemical shift
values are listed in δ scale. The FT-IR spectra were recorded on
schimadzu FT-IR spectrophotometer by using 1% potassium
bromide discs. Mass spectra of the compounds were recorded on
electronic spin ionization mass spectra (ESI-MS) on aglient 1100
series.
further for 4h at 37 °C in presence of 5% CO2. Finally, the cells were
solubilized by adding solubilizing solution and allowed to incubate
at 37 °C overnight. After complete solubilization of the formazan
crystals the absorbance was read at 540 nm in a microplate reader
(Bio-Rad, USA). The results (mean OD±SD) obtained from
quadruplicate wells were used in the calculation to determine the
cytotoxicity (50% of inhibitory concentration, IC50) of the test
compounds.
Evaluation of anti-inflammatory effect of compound 4d against
potassium oxonate induced gout in mice
Synthesis of 4-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl) benzene
sulfonamide (2)
Anti-inflammatory activity of compound 4d in potassium oxonate
induced gout in mice was investigated in the present study. Intra-
peritoneal injections of potassium oxonate 250 mg/kg daily for 28 d
has lead to increased levels of serum concentration of creatinine,
uric acid, blood urea nitrogen, and xanthine oxidase enzyme level as
an indicator of impaired kidney function and increased plasma
concentration of the blood parameters such as total leukocyte count,
differential leukocyte count and erythrocyte sedimentation rate
(ESR) indicates inflammation. Treatment with compound 4d in
doses 50 mg/kg and 100 mg/kg for 28 d exhibited a significant
improvement in gout disease in mice as evidenced by the decrease
in biochemical parameters and inflammation in joints. The total
leukocyte count, differential leukocyte count and erythrocyte
sedimentation rate were estimated as per reported methods.
Creatinine level was determined in serum by using modified jaffe’s
reaction. The level of xanthine oxidase was determined by. Uric acid
level was determined in serum by using peroxidase endpoint assay.
Blood urea nitrogen level was determined by using berthelot end
point assay. Present study indicated that the group of mice given
standard reference compound allopurinol at 5 mg/kg body weight
significantly (p<0.05) reduced serum creatinine, uric acid, blood urea
nitrogen, and xanthine oxidase enzyme level in hyperuricemic mice.
Sodium metal was dissolved in 10 ml of absolute ethanol in a dry
flask to which azido benzene sulfonamide and 2 ml of acetylacetone
were added and stirred for 1 h on magnetic stirrer. Transferred to
the round-bottomed flask (RBF) and refluxed at 140 °C for 12-16 h.
The completion of the reaction was monitored by TLC. The reaction
mixture was poured into crushed ice to obtain the solid product.
Then the precipitate was filtered under suction, washed thoroughly
with water and recrystallized from aq. methanol.
Synthesis of substituted 4-(4-(3-(phenyl)acryloyl)-5-methyl-
1H-1,2,3-triazol-1-yl)benzenesulfonamide derivatives 3(a-i)
Different substituted benzaldehydes (0.01 mol) and 4-(4-acetyl-5-
methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide 2 (0.01 mol) were
dissolved in 95% of ethanol in a conical flask to this added NaOH
pellet the reaction mixture was stirred for 30 min. The solid
obtained was collected and recrystallized from absolute ethanol.
Synthesis of substituted 4-(4-(5-(phenyl)-4,5-dihydro-1H-pyrazol-
3-yl)-5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide
derivatives 4(a-l)
Substituted chalcones 3(a-o) (0.01 mol), hydrazine hydrate (0.015
mol) were mixed in ethanol in a RBF and refluxed for 3 h. The
completion of the reaction was monitored by TLC. The reaction
mixture was poured into crushed ice to obtain the solid product.
Solid was collected and recrystallized from methanol.
Male albino mice were obtained from Sainath Agencies Hyderabad.
They were acclimatized to laboratory conditions (22±2 ˚C of
temperature, 12-hr light/dark cycle), food and water were given ad
libitum. After an acclimatization period of 2 w, they were randomly
divided into 5 experimental groups.
Synthesis of substituted 4-(5-methyl-4-(5-phenyl-4,5-dihydro-
1H-pyrazol-3-yl)-1H-1,2,3-triazol-1-yl)benzenesulfonamide
5(a-i)
To evaluate Anti-inflammatory effect of Quinoline-3-carbonitrile
derivative against potassium oxonate induced gout in mice. 30 Male
albino mice weighing 25-30 gm were randomly divided into five
groups of six mice in each group such as:
Substituted chalcones 3(a-o) (0.01 mol), substituted phenyl
hydrazines (0.01 mol) and a catalytic amount of glacial acetic acid
were refluxed in ethanol in RBF for 4-5 h After completion of the
reaction, the reaction mixture was poured into crushed ice to obtain
the solid product. Then solid was collected and recrystallized from
aq. ethanol.
Group 1: Normal control (received saline p. o daily for 28 d).
Group 2: Disease control (received potassium oxonate 250 mg/kg i.
p for 28 d).
Group 3: Standard control (received potassium oxonate 250 mg/kg
i. p and allopurinol 5 mg/kg p. o for 28 d).
Molecular docking and ADMET studies
The molecular docking study of synthesized compounds was done
by employing the Libdock protocol in order to identify binding
interactions with the target protein PI3Kα (PDB ID: 3ZIM). ADMET
studies were carried out by using discovery studio software.
Group 4: Low dose group (received potassium oxonate 250 mg/kg i.
p and Quinoline-3-carbonitrile derivative 50 mg/kg p. o for 28 d).
Group 5: High dose group (received potassium oxonate 250 mg/kg
i. p and Quinoline-3-carbonitrile derivative 100 mg/kg p. o for 28 d).
Cytotoxic activity
All the groups except normal control have received potassium
oxonate 250 mg/kg intraperitoneally for 28 d and 1 hour later,
standard group and treatment groups have received allopurinol (5
mg/kg p. o) and quinoline-3-carbonitrile (low dose-50 mg/kg and
high dose-100 mg/kg p. o) respectively. Differential leukocyte count
and total leukocyte count were estimated on 7th, 14th and 29th day by
collecting blood through retro-orbital plexus. On 29thday blood
samples were collected for estimating biochemical parameters and
later on all animals were sacrificed for histopathological studies.
The title compounds were evaluated for In vitro antiproliferative
activity via MTT ([3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium bromide]) based cytotoxic assay[31] against MCF-7
breast cancer cell line with taxol as standard reference. Cell lines
were purchased from the national institute of nutrition (NIN)
Hyderabad. Cells were harvested from the logarithmic phase of
cultures and re-suspended in Dulbecco’s Modified Eagle’s Medium
supplemented with 10% fetal bovine serum (FBS). The cell counts
were adjusted and equal number of cells were plated into each well
of 96-well culture plates and allowed to grow overnight at 37 °C, in
presence of 5% CO2. The cells were treated with test substances at
various concentrations as indicated for 72h. In vehicle control
All the experimental procedures were carried out in accordance
with the committee for the purpose of control and supervision of
experiments on animals (320/CPCSCEA dated 03-01-2001)
guidelines. The study was reviewed and approved by the
Institutional Animal Ethics Committee (GPRCP/IAEC/ 10/18/
02/PCL/AE-2-Mice-M/F-30), G. Pulla Reddy College of Pharmacy,
Mehdipatnam, Hyderabad, India.
culture wells,
a maximum of 0.5% DMSO was added. Culture
medium was renewed at every 24h with fresh culture medium
supplemented with test substances. Thereafter, 0.5 mg/ml of MTT
reagent was added to each well and the microplate was incubated
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Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
RESULTS AND DISCUSSION
Chemistry
pyrazol-3-yl)-1H-1,2,3-triazol-1-yl) benzene sulfonamides 4(a-l) (Step
4A) in good yields. Simultaneously, intermediate (3) condensed with
different substituted phenylhydrazines in ethanol under reflux for 4-
5 h to obtain 4-(5-methyl-4-(5-phenyl-4,5-dihydro-1H-pyrazol-3-
yl)-1H-1,2,3-triazol-1-yl)benzenesulfonamides of series-2 [5(a-i)]
(Step 4B) in good yields. All the derivatives were characterized by 1H
NMR, IR and ESI-MASS spectral data.
The synthesis of compounds 4(a-l) and 5(a-i) is given under fig 1. In
this scheme, the intermediate 4-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-
yl)benzenesulfonamide (2) was synthesized by diazotization of 4-
amino benzene sulphonamide and followed by condensation of 4-
azidobenzene sulphonamide with acetylacetone in presence of sodium
ethoxide under reflux for 12-15 h. Claisen Schmidt condensation
reaction of intermediate (2) with various substituted aromatic
aldehydes under basic conditions gave chalcones 3(a-l). In the final
step, reaction of chalcones 3(a-l) with hydrazine hydrate in ethanol
under reflux for 3-4 h gave 4-(5-methyl-4-(5-phenyl-4,5-dihydro-1H-
1
The H NMR spectrum of final compounds contains a singlet around
2.61 ppm for CH3 protons, protons of CH2 and CH of pyrazoline ring
appeared as doublet of doublets, triplet of doublets in the range of 3-
5 ppm, aromatic protons appeared in the range of 6.5-8.2 ppm and
two protons of–NH2 appeared as a singlet at 7.6 ppm whereas one
proton of pyrazoline NH appeared as singlet at 7.7 ppm.
Fig. 1: Synthesis of pyrazoline incorporated 1,2,3-triazole benzene sulphonamide derivatives
Spectral characterization of synthesized compounds
4-(4-(3-(4-Chlorophenyl)acryloyl)-5-methyl-1H-1,2,3-
triazol-1-yl)benzene sulfonamide (3b): Yield 97%, mp 360-
361 °C. IR spectrum, ν, cm-1: 1091, 1159, 1315, 1664, 3304,
3350. 1H NMR (400 MHz, DMSO-d6): 2.61 (s, 3H, CH3), 7.5 (d, 2H,
ArH, J= 6.9 Hz), 7.6 (d, 2H, ArH), 7.8-7.95 (m, 7H, ArH and NH2,
olefinic), 8.02 (d, 1H, olefinic). ESI-MS: m/z 403 (M+1) observed
for C18H15ClN4O3S.
4-(4-Acetyl-5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide
(2): Yield 95%, mp 129-131 °C. IR spectrum, ν, cm-1: 1174, 1357,
1689, 3192, 3273. 1H NMR (400 MHz, DMSO-d6): 2.56 (s, 3H, CH3),
2.65 (s, 3H, COCH3), 7.61 (s, 2H,-NH2), 7.88 (dd, 2H, ArH, J= 6.9 Hz),
8.08 (dd, 2H, ArH, J=9.0Hz). ESI-MS: m/z 281 (M+1) observed for
C11H12N4O3S.
4-(4-(3-(4-Methoxyphenyl)acryloyl)-5-methyl-1H-1,2,3-
triazol-1-yl)benzene sulfonamide (3c): Yield 98%, mp 260-
262 °C. IR spectrum, ν, cm-1: 1031, 1164, 1345, 1654, 3213,
3396. 1H NMR (400 MHz, DMSO-d6): 2.61 (s, 3H, CH3), 3.81 (s,
3H, OCH3), 6.9-7.1 (m, 4H, olefinic and ArH), 7.78-7.9 (m, 6H,
ArH, NH2), 8.0 (d, 2H, ArH, J= 7.0 Hz). ESI-MS: m/z 399 (M+1)
observed for C19H18N4O4S.
4-(4-Cinnamoyl-5-methyl-1H-1,2,3-triazol-1-yl)benzene
sulfonamide (3a): Yield 95%, mp 340-342 °C. IR spectrum, ν, cm-1:
1174, 1357, 1656, 3192, 3273. 1H NMR (400 MHz, DMSO-d6): 2.61 (s,
3H,-CH3), 7.51 (t, 3H, ArH), 7.6 (s, 2H,-NH2), 7.75 (d, 2H, olefinic,
ArH), 7.8-8.05 (m, 6H, ArH and NH2). ESI-MS: m/z 369 (M+1)
observed for C18H16N4O3S.
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Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
4-(5-Methyl-4-(3-(thiophen-2-yl)acryloyl)-1H-1,2,3-triazol-1-
yl)benzene sulfonamide (3d): Yield 98%, mp 350-352 °C. IR
spectrum, ν, cm-1: 1101, 1305, 1656, 3298, 3340. 1H NMR (400 MHz,
DMSO-d6): 2.61 (s, 3H, CH3), 7.2 (d, 1H, olefinic), 7.57-7.65 (m, 4H,
ArH), 7.71 (s, 2H, NH2), 7.8 (d, 1H, ArH), 7.9 (d, 2H, ArH), 8.02 (d, 1H,
olefinic). ESI-MS: m/z 375 (M+1) observed for C16H14N4O3S2.
145.2 (C-SO2). ESI-MS: m/z 461 (M+1) observed for C18H17BrN6O2S,
Anal calcd: C, 46.86; H, 3.71; Br, 17.32; N, 18.22; O, 6.94; S, 6.95; found:
C, 45.95; H, 4.71; Br, 17.22; N, 18.36; O, 6.94; S, 6.82;
4-(4-(5-(4-Methoxyphenyl)-4,5-dihydro-1H-pyrazol-3-yl)-5-
methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4f): Yield 73
%, mp 165-167 °C. IR spectrum, ν, cm-1: 1029, 1153, 1338, 1593,
2362, 3259, 3324. 1H NMR (400 MHz, DMSO-d6) 2.59 (s, 3H, CH3), 2.9
(dd, 1H, CH2), 3.58 (dd, 1H, CH2), 3.77 (s, 3H, OCH3), 4.8 (td, 1H, CH),
6.9 (d, 2H, ArH), 7.3 (d, 2H, ArH), 7.55-7.65 (m, 3H, NH2, NH), 7.9 (d,
2H, ArH), 8.13 (d, 2H, ArH). [13]C NMR (100 MHz, DMSO-d6) δ: 10.4
(CH3), 42.3 (C-Py), 55.6 (OCH3), 62.2 (C-N), 123.9, 127.5, 128.8,
128.9, 132.0, 132.3, 139.6, 142.3, 143.8 (C-Ar), 145.2 (C-SO2), 151.9
(ArC-OCH3). ESI-MS: m/z 413 (M+1) observed for C19H20N6O3S, Anal
calcd: C, 55.33; H, 4.89; N, 20.38; O, 11.64; S, 7.76; found: C, 56.33; H,
3.77; N, 21.64; O, 10.38; S, 7.88;
4-(4-(3-(Furan-2-yl)acryloyl)-5-methyl-1H-1,2,3-triazol-1-yl)
benzenesulfonamide (3e): Yield 95%, mp 210-212 °C. IR
spectrum, ν, cm-1: 1163, 1344, 1654, 3255, 3334. 1H NMR (400 MHz,
DMSO-d6): 2.61 (s, 3H, CH3), 7.2 (d, 1H, olefinic), 7.57-7.6 (m, 4H,
ArH), 7.61 (s, 2H, NH2), 7.8 (d, 1H, ArH), 7.9 (d, 2H, ArH), 8.02 (d, 1H,
olefinic). ESI-MS: m/z 359 (M+1) observed for C16H14N4O4S.
4-(4-(3-(4-(Dimethylamino)phenyl)acryloyl)-5-methyl-1H-1,2,3-
triazol-1-yl) benzenesulfonamide (3f): Yield 90%, mp 252-254 °C.
IR spectrum, ν, cm-1: 1134, 1357, 1653, 3253, 3334. 1H NMR (400 MHz,
DMSO-d6): 2.61 (s, 3H, CH3), 2.95 (s, 6H, N(CH3)2), 7.41-7.44 (m, 3H,
ArH and olefinic), 7.7-7.95 (m, 6H, ArH, NH2, olefinic), 7.96-8.15 (m,
3H, ArH). ESI-MS: m/z 412 (M+1) observed for C20H21N5O3S.
4-(5-Methyl-4-(5-(p-tolyl)-4,5-dihydro-1H-pyrazol-3-yl)-1H-
1,2,3-triazol-1-yl)benzenesulfonamide (4g): Yield 73 %, mp 210-
213 °C. IR spectrum, ν, cm-1: 1153, 1352, 1596, 2362, 3259, 3324. 1H
NMR (400 MHz, DMSO-d6) 2.59 (s, 6H,2-CH3), 2.9 (dd, 1H, CH2), 3.48
(dd, 1H, CH2), 4.77 (td, 1H, CH), 6.9 (d, 2H, ArH), 7.3 (d, 2H, ArH),
7.55-7.65 (m, 3H, NH2, NH), 7.9 (d, 2H, ArH), 8.13 (d, 2H, ArH). [13]C
NMR (100 MHz, DMSO-d6) δ: 10.4 (CH3), 21.2, 42.3 (C-Py), 62.2 (C-
N), 125.9, 127.5, 128.8, 128.9, 132.0, 132.3, 138.4, 139.6, 142.3,
143.8 (C-Ar), 145.2 (C-SO2). ESI-MS: m/z 397 (M+1) observed for
C19H20N6O2S, Anal calcd: C, 57.56; H, 5.08; N, 21.20; O, 8.07; S, 8.09;
found: C, 56.08; H, 6.56; N, 20.09; O, 9.07; S, 8.20;
4-(5-Methyl-4-(5-phenyl-4,5-dihydro-1H-pyrazol-3-yl)-1H-
1,2,3-triazol-1-yl) benzene sulfonamide (4a): Yield 69%, mp 203-
205 °C. IR spectrum, ν, cm-1: 761, 1165, 1346, 1592, 2370, 3259,
3352. 1H NMR (400 MHz, DMSO-d6):2.61 (s, 3H, CH3), 3.2 (dd, 1H,
CH2), 4.1 (dd, 1H, CH2), 5.9 (td, 1H,-CH), 7.15-7.41 (m, 5H, ArH), 7.6
(s, 2H, NH2), 7.7-7.95 (m, 3H, ArH, NH), 8.0-8.17 (m, 2H, ArH). [13]C
NMR (100 MHz, DMSO-d6): δ 10.4 (CH3), 42.3 (C-Py), 62.2 (C-N),
124.5, 125.5, 127.9, 128.4, 128.8, 132.0, 132.3, 139.6, 142.3, 145.8(C-
Ar), 145.2 (C-SO2). ESI-MS: m/z 383 (M+1) observed for
C18H17N6O2S, Anal calcd: C, 56.53; H, 4.74; N, 21.97; O, 8.37; S, 8.38,
found: C, 55.53; H, 5.38; N, 20.38, O, 8.97; S, 9.74;
4-(5-Methyl-4-(5-(4-nitrophenyl)-4,5-dihydro-1H-pyrazol-3-
yl)-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4h): Yield 75 %,
mp 205-207 °C. IR spectrum, ν, cm-1: 1174, 1357, 1595, 3262, 3373.
1H NMR (400 MHz, DMSO-d6) 2.61 (s, 3H,-CH3), 2.9 (dd, 1H,-CH2), 3.6
(dd, 1H,-CH2), 4.85 (td, 1H,-CH), 7.4 (m, 4H, ArH), 7.6 (s, 2H,-NH2),
7.7 (s, 1H,-NH), 7.9 (d, 2H, ArH), 8.1 (d, 2H, ArH). [13]C NMR (100
MHz, DMSO-d6) δ: 10.4 (CH3), 42.3 (C-Py), 62.2 (C-N), 125.9, 127.5,
128.8, 128.9, 132.0, 132.3, 139.6, 142.3, 143.8(C-Ar),, 145.2(C-SO2),
149.2 (C-NO2). ESI-MS: m/z 428 (M+1) observed for C18H17N7O4S,
Anal calcd: C, 50.58; H, 4.01; N, 22.94; O, 14.97; S, 7.50; found: C,
50.50; H, 5.58; N, 22.01; O, 14.94; S, 6.97;
4-(4-(5-(4-Chlorophenyl)-4,5-dihydro-1H-pyrazol-3-yl)-5-methyl
-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4b): Yield 78%, mp
155-158 °C. IR spectrum, ν, cm-1: 836, 1161, 1306, 1593, 2372, 3259,
3332. 1H NMR (400 MHz, DMSO-d6); 2.61 (s, 3H,-CH3), 2.9 (dd, 1H,-
CH2), 3.6 (dd, 1H,-CH2), 4.85 (td, 1H,-CH), 7.4 (m, 4H, ArH), 7.6 (s, 2H,-
NH2), 7.7 (s, 1H,-NH), 7.9 (d, 2H, ArH), 8.1 (d, 2H, ArH). [13]C NMR
(100 MHz, DMSO-d6): δ 10.4 (CH3), 42.3 (C-Py), 62.2 (C-N), 125.9,
127.5, 128.8, 128.9, 132.0, 132.3, 138.4, 139.6, 142.3, 143.8 (C-Ar),
145.2 (C-SO2). ESI-MS: m/z 417 (M+1) observed for C18H17ClN6O2S,
Anal calcd: C, 51.86; H, 4.11; Cl, 8.50; N, 20.16; O, 7.68; S, 7.69; found: C,
52.11; H, 3.86; Cl, 8.60; N, 21.16; O, 7.68; S, 6.59;
4-(4-(5-(2,4-Dimethoxyphenyl)-4,5-dihydro-1H-pyrazol-3-yl)-5-
methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4i): Yield 65
%, mp 210-212 °C. IR spectrum, ν, cm-1: 1174, 1357, 1594, 3232, 3363.
1H NMR (400 MHz, DMSO-d6) 2.61 (s, 3H,-CH3), 2.9 (dd, 1H,-CH2), 3.6
(dd, 1H,-CH2),3.9 (s, 6H, 2-OCH3), 4.85 (td, 1H,-CH), 7.4 (m, 4H, ArH),
7.6 (s, 2H,-NH2), 7.7 (s, 1H,-NH), 7.9 (d, 2H, ArH), 8.1 (d, 2H, ArH).
[13]C NMR (100 MHz, DMSO-d6) δ: 10.4 (CH3), 42.3 (C-Py), 55.6, 57.4,
62.2 (C-N), 108.2, 110.5, 120.2, 128.8, 128.9, 132.0, 132.3, 139.6, 142.3,
143.8 (C-Ar), 145.2 (C-SO2), 151.9, 153.2 (ArC-OCH3). ESI-MS: m/z 443
(M+1) observed for C20H22N6O4S, Anal calcd: C, 54.29; H, 5.01; N, 18.99;
O, 14.46; S, 7.25; found: C, 53.01; H, 6.29; N, 17.46; O, 14.99; S, 8.25;
4-(4-(5-(2-Chlorophenyl)-4,5-dihydro-1H-pyrazol-3-yl)-5-
methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4c): Yield 76%,
mp 158-160 °C. IR spectrum, ν, cm-1: 762, 1159, 1352, 1593, 2361,
3258, 3352. 1H NMR (400 MHz, DMSO-d6): 2.61 (s, 3H,-CH3), 2.9 (dd,
1H,-CH2), 3.6 (dd, 1H,-CH2), 4.85 (td, 1H,-CH), 7.4 (m, 4H, ArH), 7.6 (s,
2H,-NH2), 7.7 (s, 1H,-NH), 7.9-8.1 (m, 4H, ArH). [13]C NMR (100 MHz,
DMSO-d6): δ 10.4 (CH3), 42.3 (C-Py), 62.2 (C-N), 125.9, 127.5, 128.8,
128.9, 132.0, 132.3, 138.4, 139.6, 142.3, 143.8 (C-Ar), 145.2 (C-SO2).
ESI-MS: m/z 417 (M+1) observed for C18H17ClN6O2S, Anal calcd: C,
51.86; H, 4.11; Cl, 8.50; N, 20.16; O, 7.68; S, 7.69; found: C, 52.11; H,
3.86; Cl, 8.60; N, 21.16; O, 7.68; S, 6.59;
4-(4-(5-(4-(Dimethylamino)phenyl)-4,5-dihydro-1H-pyrazol-3-
yl)-5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4j): Yield
62 %, mp 220-222 °C. IR spectrum, ν, cm-1: 1174, 1357, 1590, 3232,
3373. 1H NMR (400 MHz, DMSO-d6) 2.61 (s, 3H,-CH3), 2.9 (s, 6H,-
N(CH3)2),3.05 (dd, 1H,-CH2), 3.6 (dd, 1H,-CH2), 4.9 (td, 1H,-CH), 7.4 (m,
4H, ArH), 7.6 (s, 2H,-NH2), 7.7 (s, 1H,-NH), 7.9 (d, 2H, ArH), 8.1 (d, 2H,
ArH). [13]C NMR (100 MHz, DMSO-d6) δ: 10.4(CH3), 25.2 (N(CH3)2),
42.3 (C-Py), 62.2 (C-N), 125.9, 127.5, 128.8, 128.9, 132.0, 132.3, 139.6,
141.2, 142.3, 143.8 (C-Ar), 145.2 (C-SO2),. ESI-MS: m/z 426 (M+1)
observed for C20H23N7O2S, Anal calcd: C, 56.45; H, 5.45; N, 23.04; O,
7.52; S, 7.54; found: C, 55.45; H, 6.45; N, 22.04; O, 8.54; S, 7.52;
4-(4-(5-(4-Fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl)-5-methyl
-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4d): Yield 95%, mp
156-158 °C. IR spectrum, ν, cm-1: 839, 1167, 1305, 1594, 2361, 3265,
3352. 1H NMR (400 MHz, DMSO-d6) 2.61 (s, 3H,-CH3), 2.9 (dd, 1H,-
CH2), 3.6 (dd, 1H,-CH2), 4.85 (td, 1H,-CH), 7.4 (m, 4H, ArH), 7.6 (s, 2H,-
NH2), 7.7 (s, 1H,-NH), 7.9 (d, 2H, ArH), 8.1 (d, 2H, ArH). [13]C NMR
(100 MHz, DMSO-d6): δ 10.4 (CH3), 42.3 (C-Py), 62.2 (C-N), 119.5,
127.5, 128.8, 128.9, 132.0, 132.3, 139.6, 142.3, 143.8 (C-Ar), 145.2 (C-
SO2). 147.5 (C-F). ESI-MS: m/z 401 (M+1) observed for C18H17FN6O2S,
Anal calcd: C, 53.99; H, 4.28; F, 4.74; N, 20.99; O, 7.99; S, 8.01; found: C,
52.28; H, 5.99; F, 5.00; N, 20.74; O, 8.99; S, 7.00;
4-(5-Methyl-4-(5-(thiophen-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)-
1H-1,2,3-triazol-1-yl)benzenesulfonamide (4k): Yield 95 %, mp
170-172 °C. IR spectrum, ν, cm-1: 1174, 1346, 1591, 3232, 3333. 1H
NMR (400 MHz, DMSO-d6) 2.58 (s, 3H, CH3), 2.95 (dd, 1H,-CH2), 3.6
(dd, 1H,-CH2), 4.85 (td, 1H, CH), 7.3 (d, 2H, ArH), 7.42-7.47 (m, 1H,
ArH), 7.6 (s, 2H, NH2), 7.7 (s, 1H, NH), 7.9 (d, 2H, ArH), 8.14 (d, 2H,
ArH). [13]C NMR (100 MHz, DMSO-d6) δ: 10.4 (CH3), 42.3 (C-Py),
62.2 (C-N), 125.9, 127.5, 128.8, 128.8, 132.0, 132.3, 138.4, 139.6,
142.3, 143.8 (C-Ar), 145.2 (C-SO2). ESI-MS: m/z 389 (M+1) observed
for C16H16N6O2S2, Anal calcd: C, 49.47; H, 4.15; N, 21.63; O, 8.24; S,
16.51; found: C, 48.15; H, 5.47; N, 20.63; O, 8.51; S, 17.24;
4-(4-(5-(4-Bromophenyl)-4,5-dihydro-1H-pyrazol-3-yl)-5-
methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (4e): Yield
72%, mp 165-167 °C. IR spectrum, ν, cm-1: 826, 1154, 1305, 1594,
2361, 3265, 3352. 1H NMR (400 MHz, DMSO-d6) 2.61 (s, 3H,-CH3), 2.9
(dd, 1H,-CH2), 3.6 (dd, 1H,-CH2), 4.85 (td, 1H,-CH), 7.4 (m, 4H, ArH), 7.6
(s, 2H,-NH2), 7.7 (s, 1H,-NH), 7.9 (d, 2H, ArH), 8.1 (d, 2H, ArH). [13]C
NMR (100 MHz, DMSO-d6) δ: 10.4 (CH3), 42.3 (C-Py), 62.2 (C-N), 124.4,
126.9, 127.5, 128.8, 128.9, 132.3, 138.4, 139.6, 142.3, 143.8 (C-Ar),
9

Siliveri et al.
Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
4-(4-(5-(Furan-2-yl)-4,5-dihydro-1H-pyrazol-3-yl)-5-methyl-1H-
123.1, 124.4, 125.1, 126.9, 127.5, 128.8, 128.9, 131.9, 132.3, 137.9,
138.4, 139.6, 142.3, 143.8 (Ar-C), 145.2 (C-SO2). ESI-MS: m/z 537
(M+1) observed for C24H21BrN6O2S, Anal calcd: C, 53.64; H, 3.94; Br,
14.87; N, 15.64; O, 5.95; S, 5.97; found: C, 53.94; H, 3.97; Br, 15.87; N,
14.64; O, 5.94; S, 5.64;
1,2,3-triazol-1-yl)benzenesulfonamide (4l): Yield 78 %, mp 190-
1
192 °C. IR spectrum, ν, cm-1: 1092, 1174, 1346, 1591, 3232, 3333. H
NMR (400 MHz, DMSO-d6) 2.58 (s, 3H, CH3), 2.95 (dd, 1H,-CH2), 3.6
(dd, 1H,-CH2), 4.85 (td, 1H, CH), 7.3 (d, 2H, ArH), 7.42-7.47 (m, 1H,
ArH), 7.6 (s, 2H, NH2), 7.7 (s, 1H, NH), 7.9 (d, 2H, ArH), 8.14 (d, 2H,
ArH). [13]C NMR (100 MHz, DMSO-d6) δ: 10.4 (CH3), 42.3 (C-Py), 62.2
(C-N), 125.95, 127.59, 128.84, 128.84, 132.09, 132.35, 138.45, 139.64,
142.34, 143.81 (C-Ar), 145.28 (C-SO2). ESI-MS: m/z 373 (M+1)
observed for C16H16N6O3S, Anal calcd: C, 51.60; H, 4.33; N, 22.57; O,
12.89; S, 8.61; found: C, 50.33; H, 5.60; N, 21.61; O, 13.89; S, 8.57;
4-(5-Methyl-4-(5-(4-nitrophenyl)-1-phenyl-4,5-dihydro-1H-
pyrazol-3-yl)-1H-1,2,3-triazol-1-yl)benzenesulfonamide (5e):
Yield 80 %, mp 154-156 °C. IR spectrum, ν, cm-1: 1114, 1357, 1592,
3192. 1H NMR (400 MHz, DMSO-d6): 2.59 (s, 3H, CH3), 2.9 (dd, 1H,
CH2), 3.6 (dd, 1H, CH2), 4.85 (td, 1H, CH), 6.95-7.15 (m, 3H, ArH),
7.18 (d, 2H, ArH), 7.4 (m, 4H, ArH), 7.6 (s, 2H, NH2), 7.7 (d, 2H, ArH),
7.9 (d, 2H, ArH). [13]C NMR (100 MHz, DMSO-d6): δ 10.4 (CH3), 42.3
(C-Py), 63.2 (C-N), 123.1, 125, 125.9, 127.5, 128.8, 128.9, 131.9,
132.0, 132.3, 137.9, 139.6, 142.3, 143.8 (Ar-C), 145.2 (C-SO2), 149.2
(ArC-NO2). ESI-MS: m/z 504 (M+1) observed for C24H21N7O4S, Anal
calcd: C, 57.25; H, 4.20; N, 19.47; O, 12.71; S, 6.37; found: C, 56.20; H,
5.25; N, 20.71; O, 12.37; S, 5.47;
4-(4-(1,5-Diphenyl-4,5-dihydro-1H-pyrazol-3-yl)-5-methyl-1H-
1,2,3-triazol-1-yl)benzenesulfonamide (5a): Yield 75 %, mp 120-
1
122 °C. IR spectrum, ν, cm-1: 1163, 1336, 1593, 3192, 3223. H NMR
(400 MHz, DMSO-d6): 2.7 (s, 3H, CH3), 3.2 (dd, 1H, CH2), 4.13 (dd, 1H,
CH2), 5.5 (td, 1H, CH), 6.7 (m, 1H, ArH), 6.95 (d, 2H, ArH), 7.15-7.25
(m, 5H, ArH), 7.26-7.7 (m, 8H, ArH, NH2). [13]C NMR (100 MHz,
DMSO-d6) δ: 10.4 (CH3), 42.3 (C-Py), 63.2 (C-N), 123.1, 125.1, 125.9,
127.5, 128.8, 128.9, 131.9, 132.0, 132.3, 137.9, 138.4, 139.6, 142.3,
143.8 (Ar-C), 145.2 (C-SO2). ESI-MS: m/z 459 (M+1) observed for
C24H22N6O2S, Anal calcd: C, 62.86; H, 4.84; N, 18.33; O, 6.98; S, 6.99;
found: C, 61.84; H, 5.86; N, 17.98; O, 7.99; S, 6.33;
4-(4-(5-(4-Methoxyphenyl)-1-phenyl-4,5-dihydro-1H-pyrazol-
3-yl)-5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (5f):
Yield 78 %, mp 125-127 °C. IR spectrum, ν, cm-1: 1106, 1343, 1600,
3233. 1H NMR (400 MHz, DMSO-d6): 2.25 (s, 3H, CH3), 3.2 (dd, 1H,
CH2), 3.78 (s, 3H, OCH3), 4.15 (dd, 1H, CH2), 5.5 (td, 1H, CH), 7.05 (m,
1H, ArH), 7.2 (d, 2H, ArH), 7.3-7.6 (m, 9H, ArH, NH2), 7.85-8.14 (m,
3H, ArH). [13]C NMR (100 MHz, DMSO-d6): δ 10.446 (CH3), 42.3 (C-
Py), 55.6 (OCH3), 63.2 (C-N), 123.1, 123.9, 125.1, 127.5, 128.8, 128.9,
131.9, 132.0, 132.3, 137.9, 139.6, 142.3, 143.8 (Ar-C), 145.2 (C-SO2),
151.9 (ArC-OCH3). ESI-MS: m/z 489 (M+1) observed for C25H24N6O3S,
Anal calcd: C, 61.46; H, 4.95; N, 17.20; O, 9.82; S, 6.57; found: C,
60.56; H, 4.20; N, 17.82; O, 9.95; S, 7.47;
4-(4-(5-(4-Chlorophenyl)-1-phenyl-4,5-dihydro-1H-pyrazol-3-yl)-
5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (5b): Yield 76
%, mp 129-131 °C. IR spectrum, ν, cm-1: 756, 1163, 1338, 1596, 3213. 1H
NMR (400 MHz, DMSO-d6): 2.7 (s, 3H, CH3), 3.2 (dd, 1H, CH2), 4.13 (dd,
1H, CH2), 5.5 (td, 1H, CH), 6.7 (m, 1H, ArH), 6.95 (d, 2H, ArH), 7.15-7.25
(m, 4H, ArH), 7.26-7.7 (m, 8H, ArH, NH2). [13]C NMR (100 MHz, DMSO-
d6): δ 10.4 (CH3), 42.3 (C-Py), 63.2 (C-N), 123.1, 125.1, 125.9, 127.5,
128.9, 129.95, 131.95, 132.09, 132.35, 137.95, 139.5, 139.6, 142.3, 143.8
(Ar-C), 145.2 (C-SO2). ESI-MS: m/z 493 (M+1) observed for
C24H21ClN6O2S, Anal calcd: C, 58.47; H, 4.29; Cl, 7.19; N, 17.05; O, 6.50; S,
6.50; found: C, 58.29; H, 4.50; Cl, 7.47; N, 16.05; O, 7.49; S, 6.20;
4-(4-(1-(4-Fluorophenyl)-5-phenyl-4,5-dihydro-1H-pyrazol-3-yl)-
5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide (5g): Yield 75
%, mp 120-122 °C. IR spectrum, ν, cm-1: 1174, 1357, 1689, 3192, 3273.
1H NMR (400 MHz, DMSO-d6): 2.65 (s, 3H, CH3), 3.15 (dd, 1H, CH2), 4.15
(dd, 1H, CH2), 5.38 (td, 1H, CH), 6.9-7.17 (m, 4H, ArH), 7.5 (d, 2H, ArH),
7.6 (s, 2H, NH2), 7.8-8.16 (m, 7H, ArH). [13]C NMR (100 MHz, DMSO-d6):
δ 10.4 (CH3), 42.3 (C-Py), 63.2 (C-N), 119.5, 123.1, 125.1, 127.5, 128.8,
128.9, 131.9, 132.0, 132.3, 137.9, 139.6, 142.3, 143.8 (Ar-C), 145.2 (C-
SO2), 147.5 (C-F). ESI-MS: m/z 477 (M+1) observed for C24H21FN6O2S,
Anal calcd: C, 60.49; H, 4.44; F, 3.99; N, 17.64; O, 6.72; S, 6.72; found: C,
61.44; H, 3.49; F, 4.64; N, 16.99; O, 6.73; S, 6.71;
4-(4-(5-(4-Fluorophenyl)-1-phenyl-4,5-dihydro-1H-pyrazol-3-
yl)-5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide
(5c):
Yield 75 %, mp 160-162 °C. IR spectrum, ν, cm-1: 832, 1163, 1332,
1600, 3233. 1H NMR (400 MHz, DMSO-d6): 2.7 (s, 3H, CH3), 3.2 (dd,
1H, CH2), 4.13 (dd, 1H, CH2), 5.5 (td, 1H, CH), 6.7 (m, 1H, ArH), 6.95
(d, 2H, ArH), 7.15-7.25 (m, 4H, ArH), 7.26-7.7 (m, 8H, ArH, NH2).
[13]C NMR (100 MHz, DMSO-d6): δ 10.4 (CH3), 42.3(C-Py), 63.2 (C-
N), 119.5, 123.1, 125.1, 127.5, 128.8, 128.9, 131.9, 132.0, 132.3,
137.9, 139.6, 142.3, 143.1 (Ar-C), 145.2 (C-SO2), 147.5 (C-F). ESI-MS:
m/z 477 (M+1) observed for C24H21FN6O2S, Anal calcd: C, 60.49; H,
4.44; F, 3.99; N, 17.64; O, 6.72; S, 6.72; found: C, 61.44; H, 3.49; F,
4.64; N, 16.99; O, 6.73; S, 6.71;
4-(4-(1-(4-Bromophenyl)-5-phenyl-4,5-dihydro-1H-pyrazol-3-
yl)-5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide
(5h):
Yield 77 %, mp 135-137 °C. IR spectrum, ν, cm-1: 856, 1163, 1338,
1596, 3213. 1H NMR (400 MHz, DMSO-d6): 2.65 (s, 3H, CH3), 3.15
(dd, 1H, CH2), 4.15 (dd, 1H, CH2), 5.38 (td, 1H, CH), 6.9-7.17 (m, 4H,
ArH), 7.5 (d, 2H, ArH), 7.6 (s, 2H, NH2), 7.8-8.16 (m, 7H, ArH). [13]C
NMR (100 MHz, DMSO-d6): δ 10.4 (CH3), 42.3 (C-Py), 63.2 (C-N),
123.1, 124.4, 125.1, 126.9, 127.5, 128.8, 128.9, 131.9, 132.3, 137.9,
138.4, 139.6, 142.3, 143.8 (Ar-C), 145.2 (C-SO2). ESI-MS: m/z 537
(M+1) observed for C24H21BrN6O2S, Anal calcd: C, 53.64; H, 3.94; Br,
14.87; N, 15.64; O, 5.95; S, 5.97; found: C, 53.94; H, 3.97; Br, 15.87; N,
14.64; O, 5.94; S, 5.64;
4-(4-(5-(4-Bromophenyl)-1-phenyl-4,5-dihydro-1H-pyrazol-3-
yl)-5-methyl-1H-1,2,3-triazol-1-yl)benzenesulfonamide
(5d):
Yield 73 %, mp 132-134 °C. IR spectrum, ν, cm-1: 840, 1153, 1340,
1600, 3233. 1H NMR (400 MHz, DMSO-d6): 2.7 (s, 3H, CH3), 3.2 (dd,
1H, CH2), 4.13 (dd, 1H, CH2), 5.5 (td, 1H, CH), 6.7 (m, 1H, ArH), 6.95
(d, 2H, ArH), 7.15-7.25 (m, 4H, ArH), 7.26-7.7 (m, 8H, ArH, NH2). 13C
NMR (100 MHz, DMSO-d6): δ 10.4(CH3), 42.3 (C-Py), 63.2 (C-N),
Table 1: Calculated docking scores, binding energies, H-bond count of the targeted compounds inside the PI3Kα active site
Compound
3j
3i
Libdock score
128.405
145.031
130.302
129.849
129.803
128.22
113.056
143.264
120.09
125.889
113.992
132.497
130.953
143.264
124.359
132.217
122.165
H-Bond count
3
4
3
2
4
1
1
1
5
3
3
3
1
2
3
0
4
3a
3e
3b
5b
4b
3d
4d
3l
4l
3f
5f
4f
3k
5i
4k
10

Siliveri et al.
Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
4-(5-Methyl-4-(1-phenyl-5-(thiophen-2-yl)-4,5-dihydro-1H-
pyrazol-3-yl)-1H-1,2,3-triazol-1-yl)benzenesulfonamide
toxicity as level 0. An ADMET plot was generated for the blood-brain
barrier penetration and the intestinal absorption using descriptors
AlogP98 and 2D polar surphase area (PSA) that comprise confidence
ellipses of 95% and 99%. These ellipses elucidate zones where well-
occupied compounds are expected to be settled. The compounds are
found to be in the range of 95 and 99 % confidence ellipse for both
the intestinal absorption and BBB as shown in fig. 4.
(5i):
Yield 73 %, mp 100-102 °C. IR spectrum, ν, cm-1: 1161, 1334, 1595,
3292. 1H NMR (400 MHz, DMSO-d6): 2.59 (s, 3H, CH3), 2.9 (dd, 1H,
CH2), 3.6 (dd, 1H, CH2), 4.85 (td, 1H, CH), 6.96-7.14 (m, 3H, ArH),
7.18 (d, 2H, ArH), 7.38 (d, 2H, ArH), 7.5 (d, 2H, ArH), 7.55-7.8 (m, 5H,
ArH, NH2). [13]C NMR (100 MHz, DMSO-d6): δ 10.446 (CH3), 42.36
(C-Py), 62.29 (C-N), 123.11, 125.11, 125.95, 127.59, 128.84, 128.84,
131.95, 132.09, 132.35, 137.95, 138.45, 139.64, 142.34, 143.81 (Ar-
C), 145.28 (C-SO2). ESI-MS: m/z 465 (M+1) observed for
C
22H20N6O2S2, Anal calcd: C, 56.88; H, 4.34; N, 18.09; O, 6.89; S, 13.80;
found: C, 56.34; H, 4.09; N, 18.89; O, 6.80; S, 13.88;
Results of docking and ADMET
Table 1 depicts the Libdock scores of top-ranked complex pose per
compound along with their binding energies and number of
hydrogen bonds. The molecular docking results of synthesized
compounds revealed that among all the docked compounds,
compound 3i possessed the high Libdock score of 145.031
compared with the reference ligand KKR exhibiting a Libdock score
of 88.35. Docking confirmation of reference ligand KKR with the
active site of protein was shown in fig 2. From the table 1, it is
revealed that all the compounds displayed convincing dock scores
ranging from 145.031 to 122.165, which implies that the designed
molecules possess a related mode of binding and valid binding
affinity against the protein just as the reference ligand. The
compound 3i is well fitted into the binding site of PI3Kα by
hydrogen bonds and other close interactions. The interaction of the
protein PI3Kα and compound 3i is shown in fig 3. From results it
was observed that four hydrogen bond interactions were presented
in the docked pose with compound 3i, two bonds with VAL851 and a
single bond with each of the residues GLN859, LYS802. N10 and N11
of the compound 3i interact with H-atom of the amine group of
VAL851 forming two hydrogen bonds (A: VAL851: HN-3i: N10 and
A: VAL851: HN-3i: N11) with a hydrogen bond distance of 1.941 Å
and 2.359 Å respectively. Third hydrogen bond was formed when
the hydrogen atom of GLN859 interacted with the oxygen atom of
the compound 3i (A: GLN859:HE22-3i: O23) with a hydrogen bond
distance of 1.882 Å. Another hydrogen bond was formed between
the hydrogen atom of LYS802 and the oxygen atom of the compound
3i (A: LYS802:HZ3-3i: O) with a hydrogen bond distance of 2.449 Å.
A few non-bonded interactions were found between the compound
3i and the residues TYR836, SER854, GLU849, VAL850, ILE932,
ILE848, ASP810, ILE800, MET772, ASP933, MET858, and CYS862.
Fig. 3: The hydrogen bond interactions of the compound 3i with
the protein PI3Kα
From the analyzed results, all the synthesized compounds showed a
BBB level of 4 except for 4k which is 3. The BBB level 4 showing
undefined penetration and level 3 indicating a little penetration
across the central nervous system (CNS) hence it reduces the side
effects linked to CNS. The absorption level was found to be 0 and 1
for all the compounds revealing good and moderate intestinal
absorption. For all the compounds, the calculated hepatotoxic level
was 1 implying the compounds as toxic. The solubility level 3
indicates very good solubility, level 2 indicates low solubility and
level
1 indicates very low solubility or no solubility. All the
compounds are found to be having the solubility level 2 except 3l,
which was 3, compound 5b was 1. Similarly, compounds having level
0 were found to be satisfactory with respect to CYP 450 2D6 liver
enzyme, suggesting that the compounds are non-inhibitors of the
metabolic enzyme and those having level 1 suggests that all the
compounds are inhibitors of the metabolic enzyme. Finally, the PPB
value found to be 2 for most of the compounds indicates that the
compounds have binding ≥ 90 % and the compounds 3i, 4l and 5k
are found to have 0 which denotes that the compounds have
binding ≤90 % clearly reveal that the compounds have good
bioavailability and are not likely to be highly bound to carrier
proteins in the blood.
Fig. 2: The docking conformation of reference drug KKR inside
the protein PI3Kα binding site
The ADMET screening results obtained for synthesized compounds
are summarized in table 2 and compared with standard levels. As
per discovery studio (DS) parameters, standard comparable values
of human intestinal absorption as level 0, solubility as level 3 and 4,
blood-brain barrier (BBB) penetration as level 3, non-inhibitory
property with cytochrome-P(CYP450 2D6) as level 0, and non-
Fig. 4: Plot of PSA versus LogP for candidate compounds
showing the 95 and 99% confidence limit ellipses
corresponding to the blood–brain barrier and intestinal
absorption models
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Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
Table 2: Predicted ADMET values of synthesised compounds
Name
BBB
level
4
4
4
4
4
4
4
4
4
4
4
4
4
4
Absorption
level
0
1
0
0
0
1
0
1
0
0
0
0
1
1
Solubility
level
2
2
2
2
2
1
2
2
2
3
2
2
2
2
Hepato
toxicity
CYP2D6
PPB
level
2
0
2
2
2
2
2
2
2
2
0
2
2
2
2
AlogP98
2D PSA
3j
3i
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
0
0
0
0
0
1
0
0
0
0
0
0
1
0
0
1
0
2.757
2.562
2.594
3.343
3.259
4.612
2.83
3.179
2.371
1.99
1.454
2.578
3.932
3.179
2.32
109.665
124.172
106.312
106.312
106.312
103.687
113.145
121.367
113.145
118.866
125.699
115.242
112.617
121.367
106.312
103.687
113.145
3a
3e
3b
5b
4b
3d
5c
3l
4l
3f
5f
4f
4k
5i
4k
3
4
4
0
0
0
2
2
2
2
0
3.567
1.785
Cytotoxic activity
Among all the tested compounds, 3b and 4d exhibited significant
percentage inhibition in cell proliferation at an IC50 value of
4.54µg/ml and 7.75µg/ml as shown in fig. 5.
Cytotoxic activity results were summarized in table 3. All tested
compounds have shown significant cytotoxic activity.
Table 3: Cytotoxicity of synthesized compounds on MCF-7 cell line
Compound IC50 value (µg/ml)
S. No.
1
2
3
4
5
6
7
3b
3f
3i
3k
3l
4d
4f
22.5
15.5
4.54
34.3
25.8
7.75
26
8
9
10
11
4k
4l
5b
19.25
55.8
43.4
15 nmol
Taxol (standard)
Fig. 5: Graphical representation of IC50 values of compounds 3i and 4d
Table 4: Effect of compound 4d on total leukocyte count (103/µl) in potassium oxonate induced gout in mice
Groups
On 7thday
On 14thday
7.862±0.729
110.0±3.443
77.80±5.001
97.02±5.100
66.46±2.954
On 29thday
7.862±0.729
120.9±7.868
24.69±0.786
62.37±5.006
13.59±1.762
Normal Control
Disease Control
Standard Control
4d (50 mg/kg)
4d(100 mg/kg)
7.862±0.729
125.0±5.510
101.7±4.728
129.5±3.521
107.1±5.094
All the data are expressed as mean±SEM (n=6).
12

Siliveri et al.
Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
Table 4.1: Effect of compound 4d on neutrophils count (103/µl) in potassium oxonate induced gout in mice
Groups
On 7th day
On 14th day
2.228±0.3582
26.73±2.672
29.48±2.917
38.07±4.850
22.16±1.875
On 29th day
Normal Control
Disease Control
Standard Control
4d (50 mg/kg)
4d (100 mg/kg)
2.228±0.3582
54.98±2.284
49.12±3.132
58.76±1.151
48.07±4.850
2.228±0.3582
10.50±0.9429
3.237±0.3677
16.35±2.999
6.305±1.037
All the data are expressed as mean±SEM (n=6).
Table 4.2: Effect of compound 4d on monocyte count (103/µl) in potassium oxonate induced gout in mice
Groups
On 7thday
On 14thday
0.18±0.043
68.06±4.847
34.19±2.433
38.73±2.883
33.69±3.270
On 29thday
0.18±0.043
97.88±6.664
19.07±1.032
29.36±2.665
3.457±0.418
Normal Control
Disease Control
Standard Control
4d (50 mg/kg)
4d (100 mg/kg)
0.18±0.043
48.57±4.362
36.72±2.663
49.16±3.324
38.73±2.883
All the data are expressed as mean±SEM (n=6).
Table 4.3: Effect of compound 4d on lymphocyte count (103/µl) in potassium oxonate induced gout in mice
Groups
On 7thday
On 14thday
5.47±0.554
14.74±1.792
13.91±1.444
19.98±2.146
10.38±0.733
On 29thday
5.47±0.554
11.93±1.313
2.338±0.406
16.35±3.332
4.910±0.5303
Normal Control
Disease Control
Standard Control
4d (50 mg/kg)
4d (100 mg/kg)
5.47±0.554
20.93±3.005
15.64±1.958
21.06±1.286
19.98±2.146
All the data are expressed as mean±SEM (n=6).
Table 4.4: Effect of compound 4d on eosinophil count (103/µl) in potassium oxonate induced gout in mice
Groups
On 7thday
0.03±0.006
0.47±0.058
0.22±0.034
0.49±0.043
0.29±0.040
On 14thday
0.03±0.006
0.47±0.067
0.21±0.036
0.19±0.040
0.21±0.030
On 29thday
0.03±0.006
0.55±0.085
0.040±0.013
0.31±0.085
0.24±0.078
Normal Control
Disease Control
Standard Control
4d (50 mg/kg)
4d 100 mg/kg)
All the data are expressed as mean±SEM (n=6)
Table 4.5: Effect of compound 4d on basophil count (103/µl) in potassium oxonate induced gout in mice
Groups
On 7thday
On 14thday
0.001±0.0004
0.023±0.008
0.023±0.006
0.037±0.011
0.018±0.002
On 29thday
Normal Control
Disease Control
Standard Control
4d (50 mg/kg)
4d (100 mg/kg)
0.001±0.0004
0.024±0.006
0.029±0.006
0.028±0.004
0.020±0.003
0.001±0.0004
0.035±0.008
0.003±0.0009
0.023±0.009
0.005±0.003
All the data are expressed as mean±SEM (n=6).
Table 5: Effect of compound 4d on erythrocyte sedimentation rate (mm/hr) in potassium oxonate induced gout in mice
Animal no
Normal control
Disease control
Standard control
Compound 4d
(50 mg/kg)
Compound 4d
(100 mg/kg)
1
1.0
8.5
2.0
3.2
2.0
2
3
2.0
0.5
5.9
9.3
4.6
2.4
4.9
6.5
4.0
1.5
4
5
2.0
1.5
10.0
8.4
3.7
4.2
7.3
6.8
2.4
1.8
6
1.0
11.0
8.850±0.7112
1.1
5.5
0.9
mean±SEM
1.333±0.2472
3.000±0.5615
5.700±0.6148
2.100±0.4320
All the data are expressed as mean±SEM (n=6).
13

Siliveri et al.
Int J Pharm Pharm Sci, Vol 11, Issue 6, 6-15
Table 6: Effect of compound 4d on xanthine oxidase (U/ml) in potassium oxonate induced gout in mice
Animal no
Normal control
Disease control
Standard control
Compound 4d
(50 mg/kg)
1.67
Compound 4d
(100 mg/kg)
1.57
1
1.69
4.94
1.94
2
3
0.59
1.59
2.29
3.01
1.40
2.23
3.36
2.36
1.62
1.91
4
5
1.22
1.64
2.16
2.31
0.78
1.67
2.04
3.05
1.37
1.45
6
0.54
1.212±0.2155
4.35
3.177±0.4860
1.52
1.590±0.2029
4.10
2.763±0.3700
2.33
1.708±0.1455
mean±SEM
All the data are expressed as mean±SEM (n=6).
Table 7: Effect of compound 4d on serum creatinine levels (mg/dl) in potassium oxonate induced gout in mice
Animal no
Normal control
Disease control
Standard control
Compound 4d
(50 mg/kg)
4.03
Compound 4d
(100 mg/kg)
1.44
1
2.5
4.69
3.21
2
3
2.0
1.5
3.38
3.23
2.94
1.06
2.89
5.23
0.96
3.50
4
5
2.5
1.0
2.61
5.30
1.72
1.39
3.12
4.27
1.70
2.40
6
1.0
6.01
4.203±0.5437
2.58
2.150±0.3598
1.98
3.587±0.4708
1.30
1.883±0.3787
mean±SEM
1.750±0.2814
All the data are expressed as mean±SEM (n=6).
Table 8: Effect of compound 4d on serum uric acid (mg/dl) in potassium oxonate induced gout in mice
Animal no
Normal control
Disease control
Standard control
Compound 4d
(50 mg/kg)
6.69
Compound 4d
(100 mg/kg)
2.68
1
2.74
9.65
5.50
2
3
2.82
2.1
11.17
11.01
5.57
5.37
6.38
6
2.94
3.62
4
5
2.54
2.34
13.05
11.44
5.43
6.43
6.56
6.86
2.99
3.07
6
1.91
2.408±0.1466
11.95
11.38±0.4579
6.06
5.727±0.1729
6.21
6.450±0.1295
3.44
3.123±0.1412
mean±SEM
All the data are expressed as mean±SEM (n=6).
Table 9: Effect of compound 4d on blood urea nitrogen (mg/dL) in potassium oxonate induced gout in mice
Animal no
1
Normal control
16.93
Disease control
32.72
Standard control
23.24
Compound 4d (50 mg/kg)
27.12
Compound 4d (100 mg/kg)
13.25
2
3
17.26
17.08
60.70
58.59
30.74
28.05
31.58
35.83
18.38
15.22
4
5
16.8
16.3
47.97
53.88
33.28
25.93
33.05
42.67
17.92
22.67
6
15.01
46.95
25.04
38.87
21.84
mean±SEM
16.56±0.3379
50.14±4.144
27.71±1.534
34.85±2.251
18.21±1.491
All the data are expressed as mean±SEM (n=6). Data analysed by one way ANOVA followed by tukeys test. αP<0.05, when compared to normal
control. βP<0.05 when compared to disease control. γP<0.05 when compared to low dose.
AUTHORS CONTRIBUTIONS
All authors had equally contributed the work
CONFLICTS OF INTERESTS
CONCLUSION
In this study, we have designed and synthesized a novel series of
pyrazoline incorporated 1, 2, 3-triazole benzene sulphonamides and
performed docking simulations in order to identify their binding affinity
towards the selected target protein PI3Kα and tested for their ADMET
profiles. Among all the compounds, compound 3i displayed preferred
binding orientations along with strong affinities towards the active site
of PI3Kα with better ADMET profile. The synthesized compounds were
examined for cytotoxic activity and compound 3i exhibited higher
activity with an IC50 value of 4.54 µg/ml. In Anti-inflammatory activity
compound, 4d has significantly shown an anti-inflammatory effect on
potassium oxonate induced gout and this was mediated by suppressing
the inflammatory responses.
Authors declare no conflicts of interest
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