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In vitro Antioxidant and Anticholinesterase Activities of Senecio massaicus

Essential Oil and Its Molecular Docking Studies as a Potential Inhibitor of
Covid-19 and Alzheimer’s Dis...

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DOI: 10.1080/22311866.2021.1955006

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In vitro Antioxidant and Anticholinesterase

Activities of Senecio massaicus Essential Oil and Its
Molecular Docking Studies as a Potential Inhibitor
of Covid-19 and Alzheimer’s Diseases

Sara Kebbi, Labib Noman, Ibrahim Demirtas, Chawki Bensouici, Sevki Adem,
Samir Benayache, Fadila Benayache, Ramdane Seghiri & Mesut Gok

To cite this article: Sara Kebbi, Labib Noman, Ibrahim Demirtas, Chawki Bensouici, Sevki Adem,
Samir Benayache, Fadila Benayache, Ramdane Seghiri & Mesut Gok (2021) In�vitro Antioxidant
and Anticholinesterase Activities of Senecio�massaicus Essential Oil and Its Molecular Docking
Studies as a Potential Inhibitor of Covid-19 and Alzheimer’s Diseases, Journal of Biologically Active
Products from Nature, 11:4, 380-394, DOI: 10.1080/22311866.2021.1955006

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 J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394 380

Original Article

In vitro Antioxidant and Anticholinesterase Activities of Senecio massaicus

Essential Oil and Its Molecular Docking Studies as a Potential Inhibitor of
Covid-19 and Alzheimer’s Diseases

Sara Kebbi 1, Labib Noman 2,3*, Ibrahim Demirtas 3, Chawki Bensouici 4, Sevki Adem 5,
Samir Benayache 1, Fadila Benayache 1, Ramdane Seghiri 1 and Mesut Gok 6

1
Unité de Recherche Valorisation des Ressources Naturelles, Molécules Bioactives et
Analyses Physicochimiques et Biologiques. Université des Frères Mentouri, Constantine,
Route d’Aïn El Bey, 25000, Constantine, Algérie
2
Department of Pharmacy, 21 September University of Medical and Applied Sciences,
Sana’a, Yemen
3
Research Laboratories Application and Research Center (ALUM), Igdir University, Igdir,
Turkey
4
Biotechnology Research Center, Ali Mendjli Nouvelle Ville UV03, BP E73, Constantine, Algeria
5
Department of Chemistry, Faculty of Science, Cankiri Karatekin University, 18100 Cankırı,
Turkey
6
Science and Technology Application and Research Center, 56210 Siirt, Turkey
*Corresponding Author: labibnomanali@gmail.com (Labib Noman)

Received 22 January 2021; Received in revised form 23 June 2021; Accepted 06 July 2021

Abstract: The composition of the essential oil obtained from the dried aerial parts of Senecio massaicus was
analyzed by GC/MS. Twenty-two components have been identified and represented 97.41 % of the total oil
composition. The major constituents of the essential oil were m-cymene (30.58 %), n-hexadecanoic acid (14.88
%) and docosane-11-decyl (10.43 %). Four methods were used to determine the antioxidant activity: DPPH,
ABTS, CUPRAC and reducing power assay. The results indicate that the essential oil extract has moderate
to low activity compared to the reference antioxidant compounds. In vitro anticholinesterase activity of the
essential oil has also been studied. It exhibited higher inhibitory activity against butyrylcholinesterase (BChE)
than against acetylcholinesterase (AChE). Docking studies conducted for Alzheimer's disease-related enzymes
have displayed that compounds docosane-11-decyl and octaethyleneglycol monododecyl ether have strong
potency, and compounds 15,15’Bi1,4,7,10,13- pentaoxacyclohexadecane and n-Hexadecanoic acid have
moderate inhibitory potential. In addition, these three compounds (Docosane-11-decyl, octaethyleneglycol
monododecyl ether and 15,15’Bi1,4,7,10,13- pentaoxacyclohexadecane) of the essential oil displayed strong
interaction against SARS-CoV-2 main protease and Nsp15 endoribonuclease. Therefore, it could be useful to
provide anticholinesterase agent and anti-coronavirus candidate drugs.
Keywords: Senecio massaicus, essential oil, COVID-19, antioxidant, anticholinesterase
Introduction caused an acute respiratory syndrome known
Since the World Health Organization’s (WHO) as COVID-19, more than 2,058,227 people have
declaration of the new corona pandemic, which died and about 95,321,880 people were infected
J. Biologically Act. Prod. Nat. 2021, 11, 380-394 © 2021 Har Krishan Bhalla & Sons
DOI:10.1080/22311866.2021.1955006
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394
with the COVID-19 in the world until 21 January
2021, and the numbers are still increasing. The
rise in deaths has become an urgent challenge
for researchers across countries to discover new
vaccines or effective drugs.
The number of COVID-19 researches related
to neuronal complications of Alzheimer’s
disease has increased recently 1, 2. The excessive
inflammation caused by COVID-19 may
accelerate the progression of brain inflammatory
neurodegeneration, which plays a main role in
AD pathology 3. Medicinal plants and their
constituents are known to have antiviral and
anticholinesterase activities 4-7. Several medicinal
herb extracts and their products were screened
as the source of natural compounds for antiviral
activities against Severe Acute Respiratory
Syndrome associated coronavirus (SARS-
CoV), which recommended some promising
compounds such as kaempferol glycosides,
alkaloids and terpenoids 2,9,10. The compounds,
which were isolated from A. annua, L. radiata,
and L. aggregata have been identified to show
antiviral activity against SARS-CoV, along
with anti-neurological inflammation and as a
candidate anti-Alzheimer's disease drugs 11-14.
Senecio genus is one of the most important and
complex genera of the Asteraceae family, which
is composed of approximately 1500 species
distributed around the world 15. Eighteen species
are present in Algeria, five of which are endemic
16
. Several species of genus Senecio have been
reported to be toxic but also for their beneficial
effects 17.
In traditional medicine, the species of the genus
Senecio belonging to the Asteraceae family have
been used for the treatment of wounds and as an
antiemetic, anti-inflammatory and vasodilatory
preparations, and as well as for the treatment
of asthma, coughs, bronchitis, stomach ache,
blood purifiers for skin eruptions and burns 18,19.
The chemical composition of the essential oil of
Senecio species has been the subject of several
studies. They revealed the presence and variation
of constituents that are exhibit biological
activities 29-23. Some Senecio species displayed
antiviral, antioxidant and anticholinesterase
activities 21,24-27. Senecio massaicus is an annual
381
herb that grows in Algeria and is collected from
the region of Bechar in the southwest of Algeria,
It also grows in Morocco.
The aim of this study was to investigate the
chemical composition of the essential oil from
Senecio massaicus and to evaluate in vitro of the
antioxidants and anticholinesterase activities,
alongside to study the molecular docking
interaction of the volatile oil components
with Covid-19 main protease and Nsp15
endoribonuclease, which are effective in virus
replication and infection in the cell, and for
Alzheimer related enzymes acetylcholinesterase
(4EY7), butyrylcholinesterase (4BDS). To the
best of our knowledge, this is the first docking
study report on the chemical composition and
biological activities of the essential oil from
Senecio massaicus.
Materials and methods
Plant material
The aerial parts of Senecio massaicus were
collected during the flowering stage from the
region of Bechar in the southwest of Algeria in
April 2017 and authenticated by Mr. Mohamed
Benabdelhakem, director of the nature pre-
servation agency, Bechar. The voucher specimen
(SM-04-2017) has been deposited in the
Herbarium of the VARENBIOMOL research
unit, Frères Mentouri University, Constantine 1.
Essential oil extraction
Dried aerial parts (150 g) of S. massaicus were
subjected to steam distillation in a Kaiser Lang
apparatus for three hours. The obtained essential
oils were collected and dried over anhydrous
sodium sulphate and kept at 4°C until analysis.
The yield of the oils was calculated in relation to
the dry weight of the plant.
GC-MS analysis
The analysis was carried out in a Trace 1310
gas chromatograph equipped with an ISQ single
quadrupole mass spectrometer (Thermo Fisher
Scientific, Austin, TX, USA). The procedure was
set to an initial temperament 60°C for 6 min, then
ramp at 4°C/min to 230°C, and finally 15 min in
230°C. The ion source and detector temperatures
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394
were 250°C and 250°C, respectively. 1 mL of
sample was dissolved in 5 mL of 100 % CHCl3
and filtered with a 0.22 µm disposable syringe
filter. A volume of 1 μL was injected in splitless
model. Separation of the sample was performed
on a Thermo TG-WAXMS GC column (60 m x
0.25 mm ID x 0.25 µm) using helium as carrier
gas at 1.2 ml/min. Mass spectral scan range was
set at the rate of 55-550 (amu). The relative %
amount of each component was calculated by
comparing its average peak area to the total area,
software adopted to handle mass spectra and
chromatograms was a Xcalibur. The retention
indexes (RI) were determined by referring to
literature (Adams, 2007). Peak identification
was conducted by comparison of the known
components stored in the NIST, Wiley7, Wiley9,
redlip, mainlip, WinRI.
Antioxidant activity
Antioxidant activity by DPPH assay
The free radical scavenging activity of the
essential oil was performed according to Blois 28.
In a 96-well plate, 160 µl of DPPH• solution was
added to 40 µl of sample solutions in methanol at
different concentrations. After incubation for 30
min in dark at room temperature, the absorbance
was measured at 517 nm. BHA and BHT were
used as antioxidant standards. The ability of the
samples to scavenge DPPH was expressed in
percent inhibition calculated using the following
equation:
DPPH ( %) = [(Acontol - Asample / Acontrol)] * 100
Where Acontrol and A sample are the absorbencies
of the reference and sample obtained from the
96-well microplate reader, respectively.
ABTS•+ radical scavenging assay
The ABTS•+ scavenging activity was tested
according to the method of Re et al. 29. The
ABTS•+ was prepared by mixing 7 mM ABTS in
water and 2.45 mM potassium persulfate, kept
in the dark at room temperature for 12 hours.
Before usage, the ABTS•+ solution was diluted
to get an absorbance of 0.703 ± 0.025 at 734
nm with ethanol or water. After that, 160 μL of
ABTS•+ was added to 40 μL of sample solution
prepared in methanol at different concentrations.
382
After incubation for 10 min the absorbance was
measured at 734 nm using a 96-well microplate
reader. BHA and BHT were used as antioxidant
standards. Percentage inhibition was calculated
for each concentration using the following
equation:
ABTS+ scavenging effect ( %) = [(Acontol - Asample
/ Acontrol)] * 100
Where Acontrol and A sample are the absorbance of
the reference and sample obtained from the 96-
well microplate reader, respectively.
Cupric reducing capacity (CUPRAC)
The cupric reducing antioxidant capacity was
determined according to the method of Apak et
al. 30. In 96-well microplate; a volume of 40 μL
of the sample was added to 50 μL of CuCl2 (10
mM). Then, 50 μL of neocuproine solution (7.5
mM) and 60 μL NH4Ac buffer (1 M, pH 7.0)
solution were given for each well. After 60 min,
the absorbance was measured at 450 nm. BHA
and BHT were used as antioxidant standards.
Results were given as A0.50 (μg /mL) which
corresponds to the concentration indicating
0.500 absorbance.
Reducing power assay
The reducing antioxidant power activity was
evaluated by Oyaizu et al. method 31. 10 μl of
sample solutions at different concentrations
were mixed with 40 μl of phosphate buffer (0.2
M, pH 6.6) and 50 μl of potassium ferricyanide
(1 %). The mixture was incubated at 50°C for
20 min, 50 μl of trichloroacetic acid (10 %)
was added. The solutions obtained were mixed
with distilled water (40 μl) and 10 μl of ferric
chloride (0.1 %), and the absorbance was
measured at 700 nm using 96-well microplate
reader. Ascorbic acid and BHA were used
as antioxidant standards. The results were
expressed as IC50 (μg mL-1) related to the
concentration, indicating 50 % absorbance
intensity.
Determination of in vitro anticholinesterase
activity
Acetylcholinesterase and butyrylcholinesterase
inhibitory activities of essential oil were
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394
measured by slightly modifying the spectro-
photometric method of Ellman et al. 32. AChE
from electric eel and BChE from horse serum
were used, while acetylthiocholine iodide and
butyrylthiocholine chloride were employed
as substrates of the reaction. 5, 50-Dithiobis
(2-nitrobenzoic) (DTNB) acid was used for
the measurement of the activity. Galantamine
was used as a positive reference compound.
The results were given as IC50 value (μg mL-1)
corresponding to the concentration shows 50 %
inhibition.
SARS-CoV-2 and anticholinesterase docking
Studies
Possible interaction mode between com-
pounds and targets were studied using the
Molegro Virtual Docker 7 33. The crystal
structures of acetylcholinesterase (4EY7),
butyrylcholinesterase (4BDS), COVID-19
virus main protease (6LU7) 34 and Nsp15
endoribonuclease (6VWW) were downloaded
from in pdb format from protein data bank (http://
www.rcsb.org/pdb). The top binding sites were
used for the docking analysis. The 3D chemical
structures of selected dietary molecules and
reference molecules were retrieved from the
PubChem site with their PubChem IDs: 6654,
440968, 519324, 22311, 7460, 10812, 30874,
5363201, 11564, 5281515, 164888, 91354,
92139, 527087, 1742210, 10219606, 10364,
560684, 123921, 41440, 985, 985. Nelfinavir was
used as a positive control for viral targets and
galanthamine for AChE and BChE.
Results and discussion
Essential oil yield and chemical composition
The chemical composition of essential oil was
analyzed by GC-MS analysis are presented in
(Table 1). The essential oil obtained by steam
distillation of aerial parts of Senecio massaicus has
an intense yellowish orange color with a specific
smell and a yield of 0.18 % (w/w). Twenty-two
compounds were identified, representing 97.41
% of total oil. This oil was characterized by the
domination of monoterpenes (51.71 %) from
which hydrocarbons monoterpenes were the most
present (45.29 %), where the main constituents
383
were m-cymene (30.58 %), n-hexadecanoic
acid (14.88 %), docosane-11-decyl (10.43 %),
α-myrcene (6.56 %), 1-indanone-2-isobutenyl-
3-methoxy (6.04 %), α-pinene (4.82 %) and
caryophyllene oxide (4.80 %). Several previous
studies on medicinal plants have been reported
that the monoterpenes contained in essential
oils are responsible for the anticholinesterase
inhibitory activity 6,8,35,36. The compounds
exist in essential oils such as α-pinene, α and
β-asarone, δ-3-carene, carvacrol 1,8-cineole,
thymohydroquinone, anethol, etc have certain
cholinesterase inhibitory activity 4.
According to the literature, there are no
reports concerning the chemical profiling of
essential oil of Senecio massaicus. Our study
shows that essential oil was characterized by
a high content of m-cymene (30.6 %). This
observation is not consistent with their reported
in previously published results on other
species. For instance, from Algeria, the major
components of S. giganteus were hexadecanoic
acid (17.8 %) followed by isophytol (12.4
%), while the same species in other studies
were α-pinene (19.4 %), 6,10,14-trimethyl-
2-pentadecanone (19.1 %) and pentacosane
(16.9 %) 23,37. Whereas γ-cadinene (15.3 %) as
the major compound of S. perralderianus 38.
From Egypt, the chief components of the oil of
S. glaucus subsp were myrcene (24.0 %) and
dehydrofukinone (I) (21.0 %) 39. In addition,
the major compounds of S. polyanthemoides
Sch. Bip. collected from two different
locations in the city of Mhlathuze, Kwa Zulu-
Natal Province were limonene (3.1-43.0 %)
and p-cymene (4.9-36.3 %) 40.
A previous study of S. leucostachys reported that
Senecio species distinguished by the prevalence
of monoterpene hydrocarbons, however, the
higher concentration components were not
m-cymene by comparison of S. aegyptius var.
discoideus, S. ambavilla, S. squalidus and S.
nutans results 41.
Moreover, the studies of other species of
Senecio from different regions have shown that
the compound m-cymene was not the major
component of the extracted oils S. flammeus,
S. nudicaulis, S. amplexicaulis, S. filaginoides,
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394 384

Table 1. Chemical composition (%) of Senecio massaicus essential oil

Peak No Retention RI a RI b Name of the compound Area %

time Literaure
1 10.58 948 945 α-Pinene 4.82
2 13.46 948 945 α-Pinene isomer 0.53
3 15.35 940 938 α-Myrcene 6.56
4 16.80 1018 1012 dl-Limonene 1.1
5 17.20 969 965 α-Phellandrene 1.70
6 19.56 1042 1037 m-Cymene 30.58
7 26.16 1294 1290 1,12-Tridecadiene 0.91
8 26.58 1324 1318 Non-3-enyl acetate 2.02
9 29.31 1290 1287 Cyclohexene-1,3-dimethyl 1.41
10 30.94 1494 1492 trans-Caryophylene 0.78
11 33.15 1131 1128 cis-Verbenol 2.34
12 34.32 825 822 Aromadendrene 1.16
13 35.92 1524 1521 α-Curcumene 1.23
14 36.87 982 978 α-Phellandrene epoxide 2.28
15 41.98 1507 1503 Caryophyllene oxide 4.80
16 46.55 937 934 Pinanediol 1.03
17 47.02 1262 1258 Carvacrol 0.77
18 47.60 8906 8903 15,15’Bi1,4,7,10,13 0.75
pentaoxacyclohexadecane
19 48.55 2628 2622 Octaethyleneglycol monododecyl ether 222
20 50.34 3138 3134 Docosane-11-decyl 10.43
21 51.46 1968 1962 n-Hexadecanoic acid 14.88
22 52.53 1738 1734 1-Indanone-2-isobutenyl-3-methoxy 6.04
a
Retention Index of references; b
Retention Index calculated from retention times relative to that of
n-alkane series

S. graveolens, S. trapezuntinus Boiss, S. expressed in IC50 μg/mL except for CUPRAC

platyphyllus var. and S. vernalis 20,22,42-47. The
climate change factors have strong effect on plant
secondary compounds 48,49. These differences in
the chemical composition of Senecio massaicus
essential oil by comparison of other species may
be due to regional variations or environmental
conditions 50.
Antioxidant activity
Antioxidant activity of the S. massaicus essential
oil was evaluated by using four different methods:
DPPH, ABTS, CUPRAC and reducing power
assay. The results presented in (Table 2) were
and reducing power methods, which were
given in terms of absorbance (A0.50 μg/mL).
The antioxidant activity of the essential oil was
compared to the standards BHA, BHT and
ascorbic acid. The essential oil exhibited higher
activity of antioxidant in reducing power and
ABTS assays with A0.50= 93.05±2.96 µg/mL and
IC50= 88.78±0.28 µg/mL respectively, compared
to CUPRAC test A0.50= 116.54±1.33 µg/mL.
For DPPH method, the essential oil didn’t have
a radical activity where the inhibition rate didn’t
exceed 37 % in the high concentration tested (200
μg/mL).
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394 385

Table 2. Antioxidant activity of essential oil of S. massaicus

Samples DPPH assay ABTS assay CUPRAC assay Reducing power assay
(IC50 µg/mL) (IC50 µg/mL) (A0.50 µg/mL) (A0.50 µg/mL)
E.O > 200 88.78±0.28 116.54±1.33 93.05±2.96
BHT a 22.32±1.19 1.29±0.30 9.62±0.87 NT
BHA a 5.73±0.41 1.81±0.10 3.64±0.19 8.41±0.67
Ascorbicacid a NT NT NT 9.01±1.46

IC50 and A0.50 values expressed are means ± SD of three parallel measurements (p < 0.05)
a
Reference compounds
NT: Not teste
Table 3. Acetylcholinesterase and butyryl-
cholinesterase inhibitory activities of the
essential oil of S. massaicus
Samples BChE AChE
(IC50 µg/mL) (IC50 µg/mL)
E.O 13.85±0.10 10.34±0.34
Galanthamine a 34.75±1.99 6.27±1.15
Values expressed are means ± S.D. of three
parallel measurements; a Reference compounds
Anticholinesterase activity
Table 3 revealed the results of acetylcholinesterase
and butyrylcholinesterase inhibitory activities
of S. massaicus essential oil, compared with
galanthamine as a positive standard. The essential
oil exhibited a potency activity close to that of
galanthamine against AChE (IC50 = 10.34±0.34
µg/mL) enzyme. In contrast, in the BChE assay,
the essential oil had higher activity than the
control with (IC50 = 13.85±0.10 µg/mL), so that
it could be useful as a good anticholinesterase
agent, especially against butyrylcholinesterase.
Alzheimer’s disease may represent a serious
reason for mortality in COVID-19 patients 2.
Therefore, S. massaicus essential oil could be used
as a possible treatment to reduce the complications
caused by SARS-CoV-2 on the nervous system.
SARS-CoV-2 and anticholinesterase docking
Studies
In order to look into the interactions between
the compounds found in the extracted oil
and the target proteins, molecular docking
of the volatile components in the active
site of the acetylcholinesterase (4EY7),
butyrylcholinesterase (4BDS), SARS-CoV-2
virus main protease (6LU7) and Nsp15
endoribonuclease (6VWW) was carried out
51
. Results presented in (Table 4) and (Table
5) express that the protein-ligand interaction
is the sum of steric effects and hydrogen bond
interactions between proteins and molecules,
while internal interaction indicates steric and
torsional effects in the small molecule. MolDock
Score is the aggregation of these two values.
When examining the docking tables and Fig. 1-4,
it can be stated that compounds has interacted
with non-polar regions the active site of enzymes.
Docosane-11-decyl, Octaethyleneglycol mono-
dodecyl ether, 15,15’Bi1,4,7,10,13 pentaoxa-
cyclohexadecane and n-Hexadecanoic acid
among the tested molecules exhibited the highest
activities against all targets. In silico results show
that these compounds play an important role in
the higher activities of the essential oil.
When comparing results to galantamine, 15,
15’Bi1,4,7,10,13 pentaoxacyclohexadecane and
n-Hexadecanoic acid demonstrated approxi-
mately the same effect as the control against
Alzheimer related enzymes. Compounds
docosane-11-decyl and octaethyleneglycol
monododecyl ether have the highest binding
affinity and bind to both the AChE and BChE
efficiently as strong inhibitors (Fig. 5). When
the in vitro and in silico results of galanthamine
against two enzymes are evaluated together, the
program can give effective results in determining
the affinity of the molecules for AChE and BChE
 Table 4. Results of the docking of essential oil compounds on the crystal
structure of Covid-19 main protease and Nsp15 endoribonuclease

Name Nsp15 endoribonuclease Mpro COVID-19

MolDock Interaction Internal HBond MolDock Interaction Internal HBond
Score Score
Nelfinavir -149.414 -171.698 22.283 -8.909 -148.413 -176.918 28.5042 -6.24452
Docosane-11-decyl -140.901 -155.934 15.033 0.000 -139.760 -156.788 17.028 0.000
Octaethyleneglycol monododecyl ether -124.917 -179.678 54.760 -4.067 -134.486 -166.117 31.630 -3.910
n-Hexadecanoic acid -111.223 -113.844 2.621 -2.522 -117.812 -121.917 4.106 -5.000
15,15’Bi1,4,7,10,13 -115.562 -113.015 -2.546 -1.845 -110.043 -107.493 -2.550 -1.430
pentaoxacyclohexadecane
1-Indanone-2-isobutenyl-3- methoxy -100.072 -93.032 -7.039 -2.686 -103.655 -98.076 -5.579 -5.000
α-Curcumene -95.781 -101.754 5.972 0.000 -99.863 -103.877 4.014 0.000
Non-3-enyl acetate -89.052 -89.215 0.164 -4.845 -97.210 -99.447 2.236 -3.714
1,12-Tridecadiene -90.832 -87.589 -3.243 0.000 -96.478 -93.759 -2.719 0.000
Caryophyllene oxide -97.588 -92.188 -5.400 0.000 -91.842 -86.441 -5.400 0.000
Aromadendrene -90.167 -87.214 -2.953 0.000 -89.990 -87.037 -2.953 0.000
trans-caryophylene -91.236 -90.011 -1.225 0.000 -81.818 -80.593 -1.225 0.000
α-Myrcene -75.070 -75.855 0.785 0.000 -80.960 -80.870 -0.090 0.000
α-phellandrene epoxide -73.467 -79.366 5.899 0.000 -72.612 -78.679 6.066 0.000
m-Cymene -67.381 -73.518 6.137 0.000 -72.599 -79.068 6.469 0.000
Carvacrol -70.624 -78.031 7.406 -2.500 -71.624 -79.166 7.541 -2.535
dl-Limonene -66.707 -72.225 5.518 0.000 -67.303 -72.624 5.321 0.000
α-Phellandrene -66.182 -74.443 8.261 0.000 -65.340 -72.702 7.361 0.000
cis-Verbenol -62.829 -79.506 16.677 -4.778 -63.105 -79.782 16.677 0.000
Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394

Cycloheexene-1,3-dimethyl -59.637 -65.073 5.436 0.000 -61.500 -66.936 5.436 0.000

α-Pinene -57.057 -72.815 15.758 0.000 -60.345 -76.103 15.758 0.000
α-Pinene isomer -57.057 -72.815 15.758 0.000 -60.344 -76.102 15.758 0.000
386

Pinanediol -60.133 -84.278 24.145 -6.409 -56.012 -80.158 24.145 -4.831

Table 5. Results of the docking of essential oil compounds on the crystal structure of Acetylcholinesterase and Butyrylcholinesterase

Name Acetylcholinesterase Butyrylcholinesterase

MolDock Protein- Internal HBond MolDock Protein- Internal HBond
Score ligand Score ligand
Interaction Interaction
Docosane-11-decyl -186.023 -213.230 27.207 0.000 -151.745 -155.155 3.410 0.000
Octaethyleneglycol monododecyl ether -171.485 -194.174 22.689 -5.207 -159.980 -205.805 45.825 -7.199
15,15’Bi1,4,7,10,13 -136.223 -132.934 -2.409 -0.881 -133.289 -130.740 -2.549 0.000
pentaoxacyclohexadecane
n-Hexadecanoic acid -128.250 -132.223 3.974 -1.033 -114.362 -113.239 1.290 2.406
Galanthamine -131.237 -136.234 4.997 -7.851 -103.729 -108.673 4.943 0.000
1-Indanone-2-isobutenyl-3- methoxy -114.774 -108.422 -6.352 -2.872 -100.360 -93.708 -6.652 -1.273
Non-3-enyl acetate -106.087 -107.620 1.533 -5.531 -99.781 -101.144 1.363 -7.981
Caryophyllene oxide -108.274 -102.874 -5.400 0.000 -95.282 -89.881 -5.400 0.000
1,12-Tridecadiene -108.062 -106.456 -1.606 0.000 -93.673 -95.545 1.872 0.000
α-Curcumene -109.415 -112.920 3.505 0.000 -92.166 -101.399 9.233 0.000
trans-caryophylene -93.544 -92.319 -1.225 0.000 -88.090 -86.865 -1.225 0.000
Aromadendrene -103.289 -100.336 -2.953 0.000 -87.864 -84.910 -2.953 0.000
α-Myrcene -77.530 -77.728 0.198 0.000 -71.805 -72.773 0.968 0.000
α-Phellandrene epoxide -72.030 -82.065 10.035 -3.012 -71.407 -77.332 5.925 0.000
Carvacrol -77.551 -85.082 7.532 -4.333 -69.864 -77.622 7.758 -2.500
m-Cymene -68.096 -74.539 6.443 0.000 -64.341 -70.421 6.080 0.000
dl-Limonene -71.647 -77.252 5.605 0.000 -63.795 -69.116 5.321 0.000
α-Phellandrene -70.194 -77.590 7.397 0.000 -63.412 -71.374 7.962 0.000
Pinanediol -59.804 -83.949 24.145 -7.104 -57.520 -81.665 24.145 -6.273
Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394

cis-Verbenol -56.496 -73.173 16.677 -2.500 -56.736 -73.413 16.677 -3.062

α-Pinene isomer -56.584 -72.342 15.758 0.000 -55.627 -71.385 15.758 0.000
α-Pinene -56.585 -72.343 15.758 0.000 -55.626 -71.384 15.758 0.000
387

Cycloheexene-1,3-dimethyl -55.906 -61.342 5.436 0.000 -54.862 -60.297 5.436 0.000

 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394 388

Figure 1. Interaction of Docosane-11-decyl, Octaethyleneglycol monododecyl ether, n-Hexa-

decanoic acid and 15,15’Bi1,4,7,10,13 pentaoxacyclohexadecane with amino acid residues of AChE

Figure 2. Interaction of Docosane-11-decyl, Octaethyleneglycol monododecyl ether, n-Hexa-

decanoic acid and 15,15’Bi1,4,7,10,13 pentaoxacyclohexadecane with amino acid residues of BChE
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394 389

Figure 3. Interaction of Docosane-11-decyl, Octaethyleneglycol monododecyl ether,

n-Hexadecanoic acid and 15,15’Bi1,4,7,10,13 pentaoxacyclohexadecane
with amino acid residues of Mpro of SARS-CoV-2

Figure 4. Interaction of Docosane-11-decyl, Octaethyleneglycol monododecyl ether,

n-Hexadecanoic acid and 15,15’Bi1,4,7,10,13 pentaoxacyclohexadecane
with amino acid residues of Nsp15 endoribonuclease
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394 390

Figure 5. Interactions of octaethyleneglycol monododecyl ether and

docosane-11-decyl with amino acid residues of AChE and BChE

Figure 6. Interactions of octaethyleneglycol monododecyl ether and docosane-11-decyl

with amino acid residues of Mpro of SARS-CoV-2 and Nsp15 endoribonuclease
enzymes. Therefore, these two compounds can viral replication and virulence. Recent studies
be to considering as candidates or lead molecules revealed that nelfinavir could be used to treat
to use in the treatment of Alzheimer’s disease. COVID–19 52. We compared our results with
For in silico viral studies, targets that we it. Docosane-11-decyl and Octaethyleneglycol
selected include the main protease SARS-CoV-2 monododecyl ether with close values to the drug
(6LU7), with the main role in the replication and exhibited a strong molecular interaction with two
transcription of the virus in the cell, and Nsp15 viral targets (Fig. 6). Also, a study reported EC50
endoribonuclease (6VWW) to mediate both values for carmofur and tideglusib as 1.82 and
 Sara Kebbi et al. / J. Biologically Act. Prod. Nat. 11 (4) 2021 pp 380 - 394 391

1.5 µM on main protease SARS-CoV-2 34. We
tested carmofur and tideglusib as the positive
control for 6LU7 using the Docking program and
obtained -115.212 and -121.345 MolDock Score.
The compounds that are actively described
above have better docking scores than we can
suggest from our results that two molecules in
the extracts may be exhibited IC50 values under
1 µM. This virus is caused by severe acute
respiratory syndrome. We think that the direct
accessibility of the compounds to the lungs
due to their volatility properties will give these
compounds an important advantage in the fight
against COVID-19.
Conclusion
The results presented in this study are the first
report on the chemical constituents and evaluation
of the antioxidant and anticholinesterase
activities of essential oil of Senecio massaicus.
The results indicate that essential oil is rich
in hydrocarbons monoterpenes (45.29 %). In
addition, the compound m-cymene (30.58 %) is
reported for the first time as a major constituent
comparing with other samples of species of the
genus Senecio. Although the essential oil showed
moderate to low antioxidant activity, it possesses
a strong capacity as an anticholinesterase
inhibitor, particularly against BChE results.
Molecular docking studies for these
enzymes have shown that Docosane-11-
decyl, Octaethyleneglycol monododecyl ether,
15,15Bi1,4,7,10,13 pentaoxacyclohexadecane,
and n-Hexadecanoic acid compounds play
an important role in the effects of extracts
on enzymes. Therefore, more in vivo tests on
Senecio massaicus essential oil are needed to
explore promising drugs. Docking studies on
two important targets in combating Covid-19
have also shown that these compounds may have
potential antiviral effects.
Acknowledgements
The authors gratefully acknowledge Lecturer
Mesut Gök from Science and Technology
Application and Research Center Siirt University,
TURKEY, for carrying out the GC-MS analysis.

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