This document describes a study that developed a micellar/niosomal vesicular nanoformulation using squalene and Tween 80 (ST8MNV) to deliver the drug naproxen (NPX) for potential anti-inflammatory and anticancer applications. The ST8MNV exhibited high encapsulation efficiency of 99.5% for NPX. In vitro tests showed the ST8MNV nanoformulation significantly reduced the half maximal inhibitory concentration of NPX in four cancer cell lines, suggesting it is a promising drug delivery system to improve the cytotoxic effects of NPX for future studies.

1. ST8 micellar/niosomal vesicular nanoformulation for delivery of
naproxen in cancer cells: Physicochemical characterization and
cytotoxicity evaluation
Vahid Erfani-Moghadam a, b, 1
, Mehrdad Aghaei c, 1
, Alireza Soltani c, *
,
Nafiseh Abdolahi c, **
, Ali Ravaghi c
, Marco Cordani d
, Shahin Shirvani c
,
Sahar Moazen Rad c
, Hanzaleh Balakheyli c
a
Medical Cellular and Molecular Research Center, Golestan University of Medical Sciences, Gorgan, Iran
b
Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran
c
Golestan Rheumatology Research Center, Golestan University of Medical Science, Gorgan, Iran
d
Institute for Advanced Studies in Nanoscience (IMDEA Nanociencia), Madrid, Spain
a r t i c l e i n f o
Article history:
Received 12 October 2019
Received in revised form
5 February 2020
Accepted 6 February 2020
Available online 13 February 2020
Keywords:
Naproxen
Niosome
Micelles
Drug delivery
Cytotoxicity
ST8MNV nanoformulation
a b s t r a c t
Naproxen (NPX) is a non-steroidal anti-inflammatory drug (NSAID) used against a variety of diseases,
including autoimmune disorders and chronic inflammations. However, low water solubility limits its
therapeutic efficacy and novel nanoformulations are required to bypass its poor bioavailability to reach
its therapeutic effect. The purpose of the study was to investigate the role of the nanoformulation of
biocompatible molecules; Squalene (S) and Tween 80 (T8) Micellar/Niosomal Vesicles (ST8MNV) pre-
pared, by thin-film hydration method and their potential as a drug delivery system for NPX. The per-
centage of encapsulation efficiency was calculated to be 99.5 ± 0.2% for 5% of NPX weight in total
ingredients of micellar/niosomal vesicles (w/w). The ST8MNV nanoformulation exhibited a slower rate of
NPX release from the drug encapsulated over seven days, suggesting a stable complex of NPX. Finally, cell
toxicity assay demonstrated that the half-maximal inhibitory concentrations (IC50) of NPX were drasti-
cally reduced by ST8MNV nanoformulation in MCF-7, A549, HeLa, and MDA-MB-231 cancer cell lines.
Our data show this micellar/niosomal naproxen nanoformulation is a great candidate for the future
in vitro and in vivo studies for potential clinical anti-inflammatory and anticancer applications.
© 2020 Elsevier B.V. All rights reserved.
1. Introduction
Non-steroidal anti-inflammatory drugs (NSAIDs) are used to
relieve pain and fever and reduce inflammation. However, some
studies reported that the use of NSAIDs increases the risk of
digestive, cardiovascular and renal complications. Their chronic
uses are usually accompanied by serious side effects such as renal,
cardiovascular and gastrointestinal complications [1,2]. Strong ev-
idence suggests that these associated NSAID therapeutic effects are
dose-dependent, since treatments with lower doses of NSAIDs may
reduce the analgesic and anti-inflammatory effect. Naproxen ((S)
-6-methoxy-a-methyl-2-naphthaleneacetic acid) (NPX) is a NSAID
used to remedy rheumatoid arthritis, osteoarthritis, chronic juve-
nile arthritis, ankylosing spondylitis, and reduced mild to moderate
pain such as headache, pain due to dysmenorrhea, postoperative
pain, and acute musculoskeletal pain [3,4]. Recent studies show
that NPX reduces the level of prostaglandins, which are responsible
for causing pain, fever, and inflammation. However, Naproxen has a
low water solubility and permeability, exhibits low absorption and
poor bioavailability which limit its therapeutic efficacy [5]. There-
fore, the development of new drug delivery systems may improve
bioavailability and thereby improve NPX pharmacokinetics.
Nanotechnology has been used in various drug production
systems to improve pharmacokinetic and pharmacodynamics
properties [6]. The low dosage treatments of nanoscale NSAIDs
were shown increasing the body drug levels, drug absorption and
* Corresponding author. Golestan Rheumatology Research Center, Golestan Uni-
versity of Medical Science, Gorgan, Iran.
** Corresponding author. Golestan Rheumatology Research Center, Golestan Uni-
versity of Medical Science, Gorgan, Iran.
E-mail addresses: alireza.soltani46@yahoo.com, alireza.soltani@goums.ac.ir
(A. Soltani), n_abdolahi2002@yahoo.com (N. Abdolahi).
1
V.E.M. and M.A contributed equally to this research.
Contents lists available at ScienceDirect
Journal of Molecular Structure
journal homepage: http://www.elsevier.com/locate/molstruc
https://doi.org/10.1016/j.molstruc.2020.127867
0022-2860/© 2020 Elsevier B.V. All rights reserved.
Journal of Molecular Structure 1211 (2020) 127867

2. represent efficient treatment by reducing undesirable side effects
[7].
Niosomes or nonionic surfactant vesicles (NSV’s) are biocom-
patible, non-immunogenic systems and have the potential to
deliver hydrophilic and hydrophobic drugs with improved chemi-
cal stability and less production cost comparing with liposomes
[8e11].
Nowadays Niosomes are broadly investigated as drug vehicles
for encapsulation and release control of different drugs such as
fluconazole [12], doxycycline hyclate [13], vancomycin [14], and
itraconazole [15]. In a recent study by Kumar and colleagues [16]
observed that by decreasing the particle size of poorly water-
soluble NSAIDs, such as ibuprofen (IBP), ketoprofen (KP), and NPX
drastically improved water solubility, anticancer properties and
declined side effects of over dosage NSAIDs.
Here, we investigated the structural conformation and appli-
cation of Squalene- Tween 80 -micellar/niosomal vesicles
(ST8MNV) prepared from the biocompatible surfactant tween 80
and the squalene, as an excipient. Interestingly, we showed that the
ST8MNV nanoformulation was efficient as drug delivery system
when strongly increase cytotoxicity of NPX in four cancer cell lines,
(MCF-7, A549, HeLa, MDA-MB-231), by significantly decreasing half
maximal inhibitory concentration (IC50) of NPX. Therefore, our
study suggests that ST8MNV nanoformulation is a potential
candidate for further anti-inflammatory and anticancer therapy
researches.
2. Material and methods
2.1. Materials
Squalene, polysorbate 80 (Tween® 80), naproxen, dicetyl
phosphate (MWCO ¼ 12,000 Da) were purchased from Sigma-
Aldrich Chemicals (Germany). Ethanol and chloroform were pur-
chased from Merck (Germany). All the reagents and solvents were
analytical grade and used without further purification.
2.2. ST8MNV preparation
ST8MNV nanoformulation preparation was done as previously
described by Erfani-Moghadam et al. [17] with some modification.
Briefly, Tween 80 and squalene were dissolved in methanol: chlo-
roform mixture at a volume ratio of 1:2 v/v and the solvent was
removed by a rotary evaporator (Rotavapor® R-114, BUCHI,
Switzerland) under low pressure. The hydration medium was
phosphate buffered saline (PBS, pH 7.4). The vesicle suspension was
sonicated (Sonopuls HD-3100, BANDELIN electronic GmbH & Co.
Germany) using a probe sonicator, in 4 cycles of 180 s “on” and 60 s
“off”, leading to the formation of uniform vesicles.
2.3. Physical properties of particles
Dynamic light scattering (DLS) and transmission electron mi-
croscopy (TEM) studies were carried out to determine the physical
properties of the ST8MNV with encapsulated naproxen. The parti-
cle size distribution (polydispersity index, PDI) of the vesicles were
dispersed in PBS (0.01 M, pH 7.4) and analyzed by DLS (Zetasizer
Nano ZS; Malvern Instruments, Malvern, UK) using an argon laser
beam at 633 nm, and a 90 scattering angle.
2.4. Encapsulation efficiency of NPX
The encapsulation efficiency (EE) of NPX was measured as
previously described by Erfani-Moghadam et al. [17]. Briefly,
different amounts of NPX (2, 5, 8, and 10 mg) and 100 mg of
lyophilized ST8MNV were co-dissolved in 3 ml of acetone in glass
tubes. Thereafter, 3 ml of water was added to each tube with stir-
ring at 35 C. Next, the acetone was evaporated and each prepared
solutions were filtered using a syringe filter (pore size: 0.22 mm) (Jet
Bio-Filtration Products, Co., Ltd., China) to remove any un-
encapsulated NPX, the obtained dispersions lyophilized, and then
10 mg of lyophilized NPX: ST8MNV were dissolved in 1 ml of
methanol to disrupt micelles/niosome structures. Samples were
then shaken vigorously for 2 min followed by 10 min of sonication
in an ultrasonic water bath. The amount of NPX in the solution was
quantified spectrophotometrically at 425 nm. Finally, the EE was
obtained by dividing the weight of NPX in nanocarrier to the weight
of feeding NPX multiplied by 100 to express the percent of drug
encapsulated in the nanocarrier.
2.5. Cell culture
Culture medium (RPMI-1640), Fetal Bovine Serum (FBS) and
Dulbecco’s PBS were purchased from Thermo Fisher Scientific
(Waltham, MA, USA). MCF-7 (human breast adenocarcinoma cell
line), A549 (human lung carcinoma), HeLa (Henrietta Lacks’
Immortal Cells), and MDA-MB-231 (human invasive ductal carci-
noma) cell lines were obtained from Pasteur Institute (Tehran, Iran).
All cell lines were cultured in (RPMI 1640) culture medium (Thermo
Fisher Scientific) supplemented with 10% FBS and
penicillinestreptomycin solution at 37 C in a humidified atmo-
sphere of 5% CO2. After the third passage, the cells were split and
used for further experiments.
2.6. Cell toxicity assay
Cell toxicity assay was performed with a tetrazolium dye MTT 3-
(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide pro-
vided by a Promega Kit (CellTiter 96⃝R AQueous One Solution Cell
Proliferation Assay). Using four different monolayer cell lines
(MDA-MDB-231, MCF-7, A549, and HeLa) were seeded at
1 Â 104
cells per well in 96-well plate in a final volume of 200 ml/
well approximately 24 h before the assay. Cells were exposed to
different concentrations (0e4000 mM) of treatments; NPX-
ST8MNV. At the end of the incubation time, 20 ml MTT reagent
(5 mg/ml in PBS) was added to the first plate (24 h) and the plate
was incubated for 2 h at 37 C in standard culture conditions.
Subsequently, the supernatant was removed, and the cells and MTT
crystals are dissolved in DMSO. The generated amount of blue
formazan is determined spectrophotometrically at 490 nm using
ELISA reader. Wells without cells serve as a negative control, and
their absorbance has to be subtracted from the other results.
2.7. Statistical analysis
Statistical analysis was performed using SPSS 19.0 software.
Results were expressed as mean ± SD. Statistical significance was
determined by Student’s t-test. P-values0.05 were considered
statistically significant.
3. Results
3.1. Encapsulation efficiency of NPX
Various amounts of NPX (2, 5, 8, and 10 mg) were loaded into
100 mg ST8MNV in dispersion and the encapsulation efficiency (EE
%) and drug loading (DL%) (Fig.1) were determined by equations (1)
and (2). The percent encapsulation efficiency was calculated to be
99.5 ± 0.2% for 5 mg of NPX weight in total ingredients weight of
the ST8MNV (w/w). Moreover, although it seems that the amount
V. Erfani-Moghadam et al. / Journal of Molecular Structure 1211 (2020) 1278672

3. of 6.6% is the maximum achieved DL (Fig. 1), the DL of about 5% for
5 mg NPX in 100 mg of ST8MNV was found to be more stable than
other ratios. These results indicated that the NPX:ST8MNV w/w
ratio of 0.05% was an appropriate ratio for the following experi-
ments due to the lower variance in EE and DL and more drug-
nanocarrier complex stability based on visual observation of the
transparency of the complex over several days.
%EE ¼ Amount of entrapped drug in the ST8
MNV=total amount of drug *100
(1)
%DL ¼ Amount of entrapped drug in the ST8
MNV=the weight of ST8MNV*100
(2)
3.2. Absorption spectra
The UVevis absorption spectra of NPX and NPX encapsulated in
ST8MNV were measured at 200e800 nm (Fig. 2). The main ab-
sorption bands for the free naproxen observed with maxima at 234,
244, 262, 272, and 332 nm, which can be assigned to the pep*
transition. The naproxen encapsulated by micellar/noisomal vesi-
cles spectrum shown in Fig. 2 has a main absorbance band around
232 nm and two weak bands at 270 and 318 nm, which can be
related to the interaction of squalene and tween 80. Zuberi et al.
have shown the main absorption peaks of naproxen are three,
around 230, 260, and 270 nm [18]. Naproxen and ST8MNV with
encapsulated naproxen display interesting photoluminescence
properties [2], the solution photoluminescent spectra of com-
pounds at room temperature have been examined (Fig. 3). It is
found that the free drug exhibit an emission upon excitation at
200 nm with maxima at 360 nm, which might be assigned to the
intraligand p-p* transition. Naproxen encapsulated by micellar/
noisomal vesicles hints blue photoluminescence with emission
peaks at 330, 350, 360, 370, and 380 nm (lex ¼ 200 nm), respec-
tively, which may be attributed to the emission of the intraligands.
3.3. FT-IR analysis
Fourier Transform Infrared (FT-IR) spectra of pure naproxen
drug and ST8MNV encapsulated naproxen have been evaluated to
study any interactions between the drug and components of the
ST8MNV. The FT-IR spectra of naproxen (data are shown in Fig. 4)
represent bands at 1428 and 1575 cmÀ1
for symmetric and asym-
metric CeC stretching vibrations. CeC stretch is observed at
1144 cmÀ1
and a sharp band at 1236 cmÀ1
is due to CeO-
stretching. The bands at 2926, 2959, and 2989 cmÀ1
can be
assigned to CeH aromatic stretching vibration [19] and at
3140 cmÀ1
because of CeH aliphatic stretching vibration [20]. The
sharp band around 1362 cmÀ1
is due to CH3 bending and the bands
at 1428 and 1453 cmÀ1
are because of asymmetrical stretching
vibration of the carboxylic acid group and at 1657 and 1701 cmÀ1
is
because of C]O symmetrical stretching vibration of the carboxylic
acid group [21,22]. The hydroxyl group (-OH) represents a band at
3181 cmÀ1
. The FT-IR spectrum of polysorbate 80 (tween 80)
showed the bands at 2916 and 2862 cmÀ1
which can be assigned to
asymmetric and symmetric vibrations of ethylene (-CH2) groups,
respectively [23]. Additionally, The FT-IR spectrum of ST8MNV
which contained tween 80 and squalene showed the bands of CH2
(2862 cmÀ1
), CH3 (2927 cmÀ1
), CeOeC (1104 cmÀ1
) and a wide
band at 3381 cmÀ1
that can be interpreted by the presence of hy-
droxyl groups [21,24]. The FT-IR spectra of complex formulation,
encapsulation of naproxen in ST8MNV, FT-IR spectra implies a
shaped band at 1150 cmÀ1
that corresponds to CeO- stretching
vibration. Two bands around 2920 and 2970 cmÀ1
can be assigned
to CeH aromatic stretching vibration (see Fig. 3). The strong
broadband at 3500 cmÀ1
could be due to the presence of OH group
of naproxen encapsulated into ST8MNV. Comparing the FTIR
spectrum of NPX encapsulated into ST8MNV with that of pure NPX
demonstrated that the characteristic band of C]O group
(1734 cmÀ1
) shifted to a high wavenumber of about 33 cmÀ1
,
suggesting formation of intermolecular hydrogen bonding can
cause significant changes in stretching vibrations between ST8MNV
and the carboxylic acid group in drug [25,26].
Fig. 1. EE and DL of NPX in ST8MNV. EE of NPX has been shown by blue line and the
left Y-axis scale and DL of NPX has been shown by red line and the right Y-axis scale.
Fig. 2. The absorption spectrum of ST8MNV, Naproxen, and Naproxen encapsulated by ST8MNV.
V. Erfani-Moghadam et al. / Journal of Molecular Structure 1211 (2020) 127867 3

4. 3.4. Physical properties of vesicles: size and morphology
Transmission electron microscopy (TEM) and dynamic light
scattering (DLS) were carried out for the naproxen encapsulated
into ST8MNV for the evaluations of the size and morphology of the
nanoformulation. TEM results demonstrated that the ST8MNV
nanoformulation was constituted by spherical vesicles in shape,
thus the morphology and characteristic size of the ST8MNV remain
unchanged after drug loading [27]. The results collected by TEM
and DLS showed that with a polydispersity index (PDI) of 0.4,
ST8MNV comprised of tween 80 and squalene had two-particle
forms, micelles with 18 ± 4.4 nm (68% of the vesicular popula-
tion), and niosome (or polymersomes) with 281 ± 80 nm (32% of
the vesicular population), both of which were spherical (Fig. 5). This
data conforms with our previous result by monomethoxy poly
[ethylene glycol]-oleate vesicles which showed the smaller vesicles
as named micelles (18.33 ± 5.32 nm) and the larger vesicles as
named polymersomes (99.4 ± 65 nm) [17]. Additionally, it has been
discussed that lowering the concentration of the surfactant leads to
an increase in the size of vesicles from micellar (10e30 nm) to
polymersome range (usually larger than 100 nm) [18,28,29].
Observing micelles beside the niosomes or polymersomes is very
common due to the thermodynamic of emulsions and surfactants
but usually neglected in scientific reports maybe because of
complexity in their interpretation. It seems that where the system
works for the determined goals it can be acceptable to have two
different classes of vesicles, micelles, and niosomes (or
polymersomes).
3.5. FESEM-EDX analysis
The field emission scanning electron microscopy with energy
dispersive X-ray (FESEM-EDX) analysis has also performed to
confirm the formation of the ST8MNV showing the presence of the
drug molecule atoms. We observed that the W% (Weight ratio
percent) was 50.14 and 43.75% whereas the A% (Atomic ratio
percent) was 43.82 and 50.93% of the ST8MNV structure. These
values correspond to the atoms of O and C that match with the
formation of the ST8MNV structure having the naproxen encap-
sulated into it (see Fig. 6).
Fig. 3. The photoluminescent spectra of Naproxen and Naproxen encapsulated by ST8MNV.
Fig. 4. The FT-IR spectra of NPX and NPX encapsulated by ST8MNV.
V. Erfani-Moghadam et al. / Journal of Molecular Structure 1211 (2020) 1278674

5. 3.6. Cell toxicity studies
We performed cell toxicity studies in several cancer cell lines to
compare the effect of NPX free and ST8MNV nanocarriers. Previ-
ously, we showed that this micellar/niosomal nanovesicles
composed of 520 mM for each of Tween 80 and squalene (520 mM:
520 mM) not only do not have significant toxicity but also has
promotional growth effect on MCF-7 and MDA-MB-231 cancer cell
lines and normal monocyte cells which can be attributed to squa-
lene, as a natural lipid [28]. Similarly, Puras et al. showed niosomes
containing squalene is safe for HEK-293, ARPE-19 cells, and for the
rat retina [30]. Here, our result showed that free NPX can kill half of
the A549 cancer cells when used at 3300 mM (mM), but when we
treated the breast cancer cells with our nanoformulation the IC50
decreased until to 400 mM (see Table 1). The significance of these
dates relies on that NPX delivered by the nanocarrier exhibits
toxicity that increased about 8.25 folds (Fig. S1). For HeLa cancer
cell lines, the increase of toxicity is about 5.5 folds and IC50 de-
creases from around 1950 to 350 mM (Fig. S1). Finally, we tested
MDA-MB-231 and MCF-7 cancer cell lines, and we report the half-
maximal inhibitory concentration (IC50) from naproxen was not
observed below the 4000 mM and again, ST8MNV formulation
indicated increased cell toxicity and IC50 were decreased to about
850 and 980 mM for MDA-MB-231 and MCF-7 cancer cell lines,
respectively (Fig. S1).
These strong differences between NPX free and NPX-ST8MNV
can be explained by its increasing of bioavailability which is
probably due to more water solubility and more efficient cell
entrance of drugs. Besides, our result corresponds to Deb et al.
which showed IC50 of three derivatives of naproxen are about 3
milimolars for MCF-7 cells. Additionally, they showed that toxicity
of naproxen sodium did not reach to the IC50 below five milimolars
for MDA-MB-231 cancer cells (IC50 ~ 10 mM) [31], which are similar
to our finding as shown in Fig. S1.
4. Discussion
Here, we showed that NPX-ST8MNV nanoformulation has more
stability and more cytotoxicity in four cancer cell lines. According to
our results, this nanoformulation potentially improves the cyto-
toxicity of NPX molecule and has the potential to increase its anti-
inflammatory effects although future studies are required in this
field. Our results are according to the previous NPX nano-
formulation studies reported in the literature. Kumar et al. used the
evaporation assisted solvent-antisolvent interaction (EASAI)
method by the means of three water-soluble and biocompatible
polymers: polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA) and
Fig. 5. TEM image and DLS of Naproxen encapsulated by ST8MNV.
Fig. 6. FESEM and EDX images of Naproxen encapsulated by ST8MNV.
Table 1
Compared IC50s of free NPX and ST8MNV with encapsulated NPX in four cancer cell
lines.
NPX (mM) NPX-ST8MNV (NPX mM)
A549 3300 400
MCF-7 Not observed below the 4000 980
MDA-MB-231 Not observed below the 4000 850
Hella 1950 350
V. Erfani-Moghadam et al. / Journal of Molecular Structure 1211 (2020) 127867 5

6. hydroxypropyl methylcellulose (HPMC) to stabilize the naproxen
nanoparticles, and they showed that nanoformulation against five
different cancer cell lines (MCF-7, MIA-PA-CA-2, HT-29, Jurkat and
A2780) improved anticancer activity against the Leukemia cancer
cell line, out of which NAP-PVP showed the highest anti-cancer
activities. The anti-Leukemia activity of NAP-PVP was more than
twice doxorubicin which is a standard anticancer drug [16]. Sharma
and colleagues have studied the effect of two anti-inflammatory
agents, naproxen and indomethacin, on the structure, assembly,
and gelation transitions of Pluronic® F127 micelles [32]. The solu-
bility of micelles was experimented with small-angle neutron
scattering methods. An ultraviolet spectroscopy evaluation sug-
gests that F127 micelles may be a better choice for drug loading
than lipid vesicles. According to the results of the study, hydro-
phobic drug solutions could reduce micellar sizes and lowering the
gelatin temperature. For efficient drug delivery, Karami and col-
leagues presented a new conjugated micellar drug carrier for nap-
roxen. Conjugation of the naproxen with diblock methoxy poly
(ethylene glycol)epoly (ε-caprolactone) (MPEGePCL) copolymer
was done through the reaction of a copolymer with naproxen in the
presence of dicyclohexylcarbodiimide and dimethylaminopyridine.
Biocompatibility of the designed drug delivery systems was tested
through cell viability assay and did not show any significant cyto-
toxic effects. In comparison with the conjugated copolymer, con-
jugated micelles had a more sustained release profile. Their result
indicated that the naproxen conjugated micelles have a good po-
tential by improvements in enhanced permeability and retention
(EPR) effect, and pH sensitivity to increase the drug release and
accumulation inside inflammatory sites [33].
5. Conclusions
In this study, we evaluated the ST8MNV nanoformulation of
naproxen, containing squalene and tween 80 which has great
similarity to the formulation of MF59®-adjuvanted influenza vac-
cine (developed by Novartis), tested in millions of people without
significant safety and immunogenicity concerns [34]. Therefore,
this ST8MNV nanoformulation is a great candidate for the future
in vitro and in vivo studies and may be of interest to evaluate the
potential clinical anti-inflammatory and anticancer applications.
Declaration of competing interest
The authors declare that they have no conflict of interest.
Acknowledgements
We gratefully acknowledge financial support from Golestan
University of Medical Science (Research Project Grant No.
970308037). We also thank the Clinical Research Development Unit
(CRDU), Sayad Shirazi Hospital, Golestan University of Medical
Sciences, Gorgan, Iran. We are deeply grateful to Dr. Ayyoob
Khosravi and Ms. Motahareh Heydari for their useful technical help.
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.molstruc.2020.127867.
Author contributions section
NA designed the study and conceived the experiments with the
help of AS and analyzed the results and prepared the manuscript
with the help of VAM, MC, and MA. AS and AR conducted the ex-
periments. SMR, SS, and HBK helped in cancer cell line experiments
and their analysis. All authors contributed toward data analysis,
drafting and critically revising the paper, gave final approval of the
version to be published, and agree to be accountable for all aspects
of the work.
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