Newly synthesized (R)-carvone-derived 1,2,3-triazoles: structural, mechanistic, cytotoxic and molecular docking studies

Article abstract of DOI:10.1080/07391102.2021.1894984

In the current study, natural (R)-carvone was used as starting material for the efficient synthesis of several 1,2,3-triazole derivatives. The produced products were obtained in good yields and characterized by ¹H and ¹³C NMR and HRM

Full text of DOI:10.1080/07391102.2021.1894984

Journal of Biomolecular Structure and Dynamics
ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/tbsd20
Newly synthesized (R)-carvone-derived 1,2,3-
triazoles: structural, mechanistic, cytotoxic and
molecular docking studies
Mouhi Eddine Hachim, Ali Oubella, Said Byadi, Mourad Fawzi, Yassine
Laamari, Lahoucine Bahsis, Aziz Aboulmouhajir, Hamid Morjani, Črtomir
Podlipnik, Aziz Auhmani & My Youssef Ait Itto
To cite this article: Mouhi Eddine Hachim, Ali Oubella, Said Byadi, Mourad Fawzi, Yassine
Laamari, Lahoucine Bahsis, Aziz Aboulmouhajir, Hamid Morjani, Črtomir Podlipnik, Aziz Auhmani
& My Youssef Ait Itto (2021): Newly synthesized (R)-carvone-derived 1,2,3-triazoles: structural,
mechanistic, cytotoxic and molecular docking studies, Journal of Biomolecular Structure and
Dynamics, DOI: 10.1080/07391102.2021.1894984
To link to this article: https://doi.org/10.1080/07391102.2021.1894984
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JOURNAL OF BIOMOLECULAR STRUCTURE AND DYNAMICS
https://doi.org/10.1080/07391102.2021.1894984
Newly synthesized (R)-carvone-derived 1,2,3-triazoles: structural, mechanistic,
cytotoxic and molecular docking studies
Mouhi Eddine Hachima , Ali Oubellab, Said Byadic , Mourad Fawzib, Yassine Laamarib, Lahoucine Bahsisd
a,c
e
f
, Aziz Auhmanib
and My Youssef Ait Ittob
ꢀ
, Aziz Aboulmouhajir , Hamid Morjani , Crtomir Podlipnik
aꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Equipe de modelisation moleculaire et de spectroscopie, Faculte des sciences, Universite de Chouaïb Doukkali, El Jadida, Morocco;
b
ꢁ
ꢁ
ꢂ
ꢁ
Departement de Chimie, Faculte desSciences, Laboratoire de Synthese Organique et Physico-Chimie Moleculaire, Marrakech, Morocco;
cꢁ
ꢂ
ꢁ
Equipe de spectroscopie d’extraction et de valorisation, Synthese organique, Laboratoire d’extraction et de valorisation, Faculte des
d
ꢁ
ꢁ
sciences d’Ain Chock, Universite Hassan II, Casablanca, Morocco; Laboratoire de Chimie de Coordination et d’Analytique, Departement de
Chimie, Faculte des Sciences d’El Jadida, Universite Chouaïb Doukkali, El Jadida, Morocco; BioSpectroscopie Translationnelle, BioSpecT -
EA7506, UFR de Pharmacie, Universite de Reims Champagne-Ardenne, Reims Cedex, France; Faculty of Chemistry and Chemical
e
ꢁ
ꢁ
f
ꢁ
Technology, University of Ljubljana, Ljubljana, Slovenia
Communicated by Ramaswamy H. Sarma
ABSTRACT
ARTICLE HISTORY
Received 17 December 2020
Accepted 20 February 2021
In the current study, natural (R)-carvone was used as starting material for the efficient synthesis of sev-
eral 1,2,3-triazole derivatives. The produced products were obtained in good yields and characterized
by 1H and 13C NMR and HRMS analysis. The newly synthesized monoterpenic 1,2,3-triazole 4 and
derivatives were analyzed by viability tests (MTT) for their cytotoxic activity against three human can-
cer cells. Product 5 showed a medium antitumor activity, for which the IC50 values against selected
cells HT-1080, A-549 and MCF-7 were 29.25 lM, 31.62 lM and 26.02 lM, respectively. The regioselectiv-
ity of the condensation reaction and the molecular structure of the title compounds were determined
by Density Functional Theory (DFT) using B3LYP calculations at 6-311 þ G(d,p) level. The orbitals
HOMO and LUMO were studied to determine the electronic properties of the synthesized compounds.
In addition, the global reactivity indices were used to explain the regioselectivity for the formation of
compound 6, and the theoretical results agree well with the experimental results. Molecular docking
and molecular dynamics confirmed the empirical test results and confirmed the stability of the com-
plex during inhibition of the anti-apoptotic protein for killing cancer cells.
KEYWORDS
(R)-Carvone; 123-triazole;
Click chemistry;
Regioselectivity; DFT
calculation;
Molecular docking
1. Introduction
various diseases (Newman et al., 2000). In this regard, the ter-
penic derivatives occurring from natural sources play an
increasingly important role as therapeutic and prophylactic
The naturally occurring compounds play a significant role in
various applications, such as drug designing and remedies of
agents
for
human
diseases
(Brahmkshatriya
&
ꢁ
ꢁ ꢁ ꢁ
Equipe de modelisation moleculaire et de spectroscopie, Faculte des sciences,
CONTACT Mouhi Eddine Hachim
mouhieddinehachim@gmail.com
ꢁ
ꢁ
Departement de
Universite de Chouaïb Doukkali, El Jadida, Morocco; Ali Oubella oubellaali1@gmail.com; My Youssef Ait Itto
aititto@uca.ac.ma
ꢁ
ꢂ
ꢁ
Chimie, Faculte desSciences, Laboratoire de Synthese Organique et Physico-Chimie Moleculaire, Marrakech, Morocco
Supplemental data for this article can be accessed online at https://doi.org/10.1080/07391102.2021.1894984.
ß 2021 Informa UK Limited, trading as Taylor & Francis Group

2
M. E. HACHIM ET AL.
Brahmkshatriya, 2013). One of the important terpenic deriva- CDCl3 signal of the solvent (7.26 ppm and 77.16 ppm). The
tives is monoterpenes (C10H16) compounds, which are the
simplest type of terpenes and are present in varying
amounts in essential oils (Crowell, 1999). These monoter-
penes were found to be excellent antitumor candidates by
preventing the formation or the progression of cancer and
also by their ability to reduce existing malignant tumors
(Crowell, 1999). Heterocycles have been extensively devel-
oped and are still developing (Zimmermann et al., 2007;
Mansouri et al., 2019; Mansouri et al., 2020). For instance,
heterocyclic derivatives based on monoterpenes, in particular
the (R)-carvone, have been increasingly investigated due to
their diverse applications, especially in biology as anti-cancer
agents (Oubella et al., 2019; Oubella et al., 2020). Indeed,
there is no denying that pentagonal heterocyclic compounds
are of great interest in chemical and biological areas; in par-
ticularly heterocyclic systems combining 1,2,3-triazole or thia-
zolidinone nuclei with a terpenic skeleton, have shown a
broad spectrum of bioactivities such as antitumoral (Taia
et al., 2020) and antibacterial activities (Ousidi et al., 2016).
As part of our ongoing research strategy aiming at the
preparation and theoretical study of heterocyclic systems
with monoterpenic skeleton (Feddouli et al., 2004; Feddouli
HRMS were determined using
spectrometer.
a
Q-TOF micro mass
2.2. Procedure for the synthesis of compound 6
2.2.1. General procedure for the preparation
of compound 2
During 30 min at 25 ꢀC, 40 mL of aqueous NaOCl (5.2%) was
added dropwise to a stirred solution of (R)-carvone 1
(2 mmol) and Fe(Cl3),10H2O (3 mmol) in 50 mL of CH2Cl2/H2O
(1/1). After 30 min stirring (TLC monitored the reaction) at
room temperature, the layers were separated, and the aque-
ous layer was extracted with CH2Cl2 (3 ꢁ 10 mL). The com-
bined organic extracts were dried over anhydrous Na2SO4
and evaporated in vacuo. The residue was purified by
column chromatography using hexane/ethyl acetate mixture
(95:5) as eluent.
2.2.2. General procedure for the preparation
of compound 3
4 mmol of compound 2, 10 mL of toluene and 5 mmol of
sodium azide are placed in a round-bottomed 100 mL flask
and heated with stirring for 2 h (130 ꢀC). The mixture was
concentrated under reduced pressure and extracted with
dichloromethane (3 ꢁ 10 mL). The combined extracts were
dried over anhydrous Na2SO4, the volatiles were evaporated,
and the residue was purified via column chromatography on
silica gel using hexane/ethyl acetate mixture (97:3) as
an eluent.
ꢁ
ꢁ
et al., 2006; Oubella et al., 2020; Rıos-Gutierrez et al., 2020;
Mansouri et al., 2020), we present in this work the hemisyn-
thesis of new 1,2,3-triazoles, using 1,3-dipolar cycloaddition
reaction of azido carvone 2 (prepared in two steps from
(R)-carvone) with dimethyl acetylenedicarboxylate (DMAD),
followed by an hetero-cyclization reaction of a triazolic thio-
semicarbazone with ethyl bromoacetate. The molecular struc-
ture determination of the desired products was carried out
by NMR (1H & 13C) and HRMS analysis. The evaluation of
their cytotoxic activity against HT-1080 fibrosarcoma, lung
carcinoma A-549, and breast adenocarcinoma MCF-7 cells is
also reported here. The HOMO, LUMO and NBO analyses
were performed using DFT/B3LYPmethod at 6-311 þ G(d,p)
level. The local reactivity indexes were also carried out to
investigate the plausible mechanisms for the regioselective
synthesis of the compound 6. We used molecular docking to
evaluate our compounds interaction with one of the critical
anti-apoptotic protein, which plays a crucial role in combat-
ing cancer cells; Molecular dynamic was used to evaluate the
stability of the complexes formed by LIGAND-Bcl-2.
2.2.3. General procedure for the preparation of 1,2,3-tri-
azole-carvone 4
A solution of azido carvone 3 (1 mmol) and dimethyl acetyle-
nedicarboxylate (DMAD) (1.5 mmol) in toluene (20 mL) was
refluxed for three hours (monitoring by TLC) then concen-
trated under reduced pressure. The resulting residue was
purified by column chromatography using Hexane/Ethyl acet-
ate 40:60 as eluent.
Dimethyl(R)-1-(2-(4-methyl-5-oxocyclohex-3-en-1-yl)ally
l)-1H-1,2,3-triazole-4,5-dicarboxylate (4)
Yield 69%; yellow solid: mp ¼ 146-148 ꢀC (Ethanol). HRMS
(TOF-MS ESþ) (m/z): found 334.1349 [M þ H]þ, calculated
334.1343. 1H NMR d (ppm): 1.73 (3H, s); 2.00-3.25 (5H, m);
3.90 (3H, s); 3.99 (3H, s); 4.37 (2H, s); 5.15 (2H, 2 s); 6.32 (1H,
m). 13C NMR d (ppm): 17.97 (CH3); 29.57 (CH2); 30.02 (CH);
40.98 (CH2); 49.71 (CH3); 51.36 (CH2); 51.86 (CH3); 110.95
(H2C¼); 125.07 (C); 132.02 (C); 132.84 (C); 141.20 (C); 145.04
(¼C); 160.02 (C); 160.44 (C); 198.78 (C).
2. Experimental
2.1. Materials and methods
All chemicals and solvents required for the synthesis of com-
pound 6 were obtained from Merck and Sigma-Aldrich. Thin-
layer chromatography (TLC) was performed with F254 perco-
lation plates (0.25 mm) and visualized by U.V. fluorescence.
Column chromatographic purifications were performed on
Silica Gel 60 (230-400 mesh) using hexane/ethyl acetate mix-
tures as eluants. 1H and 13C NMR spectra were recorded at
500 and 125 MHz on Bruker Avance III spectrometer with a
BBFO probe at room temperature using Chloroform-d
(CDCl3) as the solvent. The chemical shifts (d) are given in
2.2.4. General procedure for the preparation of 1,2,3-tri-
azole-carvone thiosemicarbazone 5
Compound 5 was prepared from an equimolecular quantity
of (R)-1,2,3-triazole-carvone 4 and thiosemicarbazide dis-
solved in ethanol. The mixture was heated under reflux for
parts per million (ppm). They were recorded relative to the four hours (the reaction was monitored by TLC) in the

JOURNAL OF BIOMOLECULAR STRUCTURE AND DYNAMICS
3
2.3. Cell culture
The human fibrosarcoma cell line HT-1080 (CCL 121) was
purchased from Sigma Aldrich (ECACC collection, Saint-
Quentin Fallavier, France). The human breast adenocarcin-
oma MCF-7 (HTB-22) and lung carcinoma A-549 (CCL-185)
cell lines were purchased from the American Type Culture
Collection (ATCC). Cells were cultured in MEM with Earle salts
and Glutamax I (Invitrogen, Cergy-Pontoise, France) supple-
mented with 10% fetal bovine serum, (Invitrogen) and 1%
penicillin-streptomycin (Invitrogen). Cultures were maintained
at 37 ꢀC in a humidified atmosphere containing 5% CO2 (v/v).
Cells were routinely passaged at preconfluency using 0.05%
trypsin, 0.53 mM EDTA (In vitro gen) and screened for the
absence of mycoplasma using PCR methods.
Figure 1. Ligand-binding pocket prediction for the Bcl-2 protein using Yasara
software for visualization.
2.4. Cytotoxicity assay
presence of few drops of concentrated H2SO4. Evaporation of
the solvent under reduced pressure gave the corresponding
1,2,3-triazole-carvonethiosemicarbazone 5 which was subse-
quently purified by column chromatography using hexane/
Ethyl acetate (70:30) as eluent.
Dimethyl(R,E)-1-(2-(5-(2-carbamothioylhydrazineylidene)-
4-methylcyclohex-3-en-1-yl)allyl)-1H-1,2,3-triazole-4,5-
dicarboxylate (5)
Cytotoxicity of the synthesized compounds was assessed by
R
V
the CellTiter 96 Non-Radioactive Cell Proliferation Assay
(MTT) (Promega, Charbonnieres les Bain, France). Brie fly, the
cells were plated at a density of 2500 cells/well in a 96-well
plate with 100 mL culture medium per well, containing differ-
ent concentrations of the previously cited molecules (6.25,
12.5, 25, 50 and 100 mM). After 24 h, 15 mL of MTT dye solu-
tion was added in each well. The plates were further incu-
bated for 4 h. Then, 100 mL of the solubilization/stop solution
was added into each well, and the plate was incubated 1 h
at room temperature. The optical density of each well was
measured at 570 nm using a microplate reader Revelation
96-well multi scanner (Dynex Technologies, Chantilly, VA).
Results were represented as a percentage of cell survival
concerning untreated controls. The IC50 was defined as the
drug concentration required for inhibition of cell growth by
50%, relative to untreated controls. IC50 values were esti-
mated from the dose-response curve.
Yield 76%; white solid: mp ¼ 110-112 ꢀC (Ethanol). HRMS
(TOF-MS ESþ) (m/z): found 407.1452 [M þ H]þ, calculated
407.1439. 1H NMR d (ppm): 1.75 (3H, s); 2.00-3.50 (5H, m);
4.10 (3H, s); 4.17 (3H, s); 4.33 (2H, s); 4.90 (2H, 2 s); 6.21 (1H,
m); 6.16 (NH2, s); 9.62 (NH, s). 13C NMR d (ppm): 18.63 (CH3);
29.87 (CH2); 30.06 (CH); 39.95 (CH2); 49.82 (CH3); 51.84 (CH2);
52.03 (CH3); 110.82 (H2C¼); 125.10 (C); 133.26 (C); 135.28 (C);
141.81 (C); 144.92 (¼C); 155.10 (C); 160.13 (C); 160.24 (C);
175.14 (C).
2.2.5. General procedure for the preparation of 1,2,3-tri-
azole thiazolidinone 6
A mixture of thiosemicarbazone 5(1 mmol), ethyl 2-bromoa-
cetate (1.2 mmol) and anhydrous sodium acetate (3.6 mmol) 2.5. Computational methods
in absolute ethanol (30 mL) was stirred under reflux until the
The calculations for DFT were performed using the B3LYP
completion of the reaction (1-3 h), TLC monitored the pro-
gress of the reaction. The reaction mixture was left to cool,
poured into cold water, extracted with ethyl acetate and
purified by column chromatography (SiO2, hexane/ethyl acet-
ate gradient).
Dimethyl1-(2-((R,E)-4-methyl-5-(((E)-4-oxothiazolidin-2-yli-
dene)hydrazineylidene)cyclohex-3-en-1-yl)allyl)-1H-1,2,3-
triazole-4,5-dicarboxylate (6)
method with 6-311 Gþ(d,p) basis sets with Gaussian 09 soft-
ware (Frisch et al., 2009). The optimization of geometries has
been performed at the B3LYP/6-311Gþ(d,p) level. All
obtained frequencies are positive, proving that all structure
corresponds to minimum energy (Singh et al., 2017). The
frontier molecular orbitals, HOMO and LUMO, of the com-
pounds 4, 5 and 6 are calculated theoretically at the same
level. The electronic chemical potential (l) and chemical
hardness (g) are calculated using energies of the frontier
molecular orbital, EHOMO and ELUMO, by the following expres-
sions m¼ (EHOMO þ ELUMO)/2 and g¼ (ELUMO - EHOMO),
respectively. The global electrophilicity (x) and nucleophilic-
ity (N) indexes were measured at the same level, and are
given by the following simple expressions: x ¼ l2/2g;
N ¼ EHOMO ꢂ EHOMO (tetracyanoethylene (TCE)) (Domingo,
2014; Domingo et al., 2016; Geerlings et al., 2003).
Yield 80%; white solid: mp ¼ 154-156 ꢀC (Ethanol). HRMS
(TOF-MS ESþ) (m/z): found 447.1481 [M þ H]þ, calculated
447.1484. 1H NMR d (ppm): 1 H NMR d (ppm): 1.75 (3H, s);
2.00-3.25 (5H, m); 3.77 (2H, s); 4.09 (3H, s); 4.19 (3H, s); 4.67
(2H, s);5.24 (2H, 2 s); 6.24 (1H, m); 9.50 (NH, s).13C NMR d
(ppm): 16.36 (CH3); 29.17 (CH2); 30.10 (CH2); 32.50 (CH); 42.59
(CH); 49.77 (CH2); 52.44 (CH3); 52.74 (CH3); 112.21 (H2C¼);
127.10 (C); 131.34 (C); 134.19 (C); 134.92 (C); 142.31 (¼C);
157.13 (C); 161.20 (C); 165.04 (C); 166.31 (C); 174.28 (C).

4
M. E. HACHIM ET AL.
Scheme 1. The different derivative of the (R)-carvone synthesized in this work
Scheme 2. General schematic for the synthesis of the new thiazolidinone-1,2,3-triazolehybrid 6.
Table 1. NMR Characteristic signals of carvone derivatives 2 and 3 compared with literature.
H3C(1)
HC(2)5C
H2C(3)
H2C(4)
1
1
1
1
d H – d 13C
d H–d 13C
d H – d 13C
d H–d 13C
2
1.74 –15.65
1.76 –15.68
1.74 – 15.08
1.73 – 15.10
6.71 –144.22
6.73 –144.19
6.77 –144.81
6.74 –144.70
4.01– 46.81
4.06– 46.96
3.81– 53.77
3.87– 53.80
5.05-5.27–116.04
5.03-5.23–115.19
5.13-5.13 –113.59
5.09-5.15 –113.40
2 (Weinges & Schwarz, 1993)
3
3 (Galstyan et al., 2018)
Table 2. NMR Characteristic signals of cycloadducts 4, 5 and 6.
The general structure of synthesized products
H3C-O C4’ & C5’ H2C3 H2C4
1
1
1
d H – d 13C
d
13C
d H – d 13C
d H–d 13C
4
5
6
3.75-4.00 – 50.3-54.8
3.77-3.90 – 51.3-53.6
3.76-3.85 – 50.2-53.2
123.4 – 134.2
122.8 – 134.1
123.2 – 135.1
4.37 – 51.3
4.33 – 51.8
4.70 – 49.7
5.10-5.20 – 110.9
4.75-5.00 – 110.8
5.25.5.75 – 112.2

JOURNAL OF BIOMOLECULAR STRUCTURE AND DYNAMICS
5
Maestro software (Moitessier et al., 2008). The molecular
docking was done with Autodock Vina using Yasara software
€
(Turkes¸, 2019; Kilic et al., 2020; Sever et al., 2020). The active
sites were parameterized to be in the 3.5 Å from the co-crys-
tallized ligand’s geometric centroid (Kalaycı et al., 2021).
Ligand docking protocols generated ten different poses for
each filtered molecules by maintaining the docking tolerance
at 0.25. During docking, the scoring function and binding
interaction for every single conformational pose were chosen
as a selection criterion (Fei et al., 2013). The 3 D structure of
the Anti-apoptotic Bcl-2 protein used in our study is pre-
sented in Figure 1, we marked the binding pocket to show
the cavity and how it looks.
2.7. Molecular dynamics
Molecular dynamics (M.D.) simulation were used to analyze
the dynamic behavior of the crystalline structures of the
human anti-apoptotic Bcl-2 protein, which is very important
for the phenomenon of induction of apoptosis at cell death,
selected from Protein Data Bank (www.rcsb.org) and the
selected organic compound after molecular docking. The
complexes consisting of the human Bcl-2 protein and three
compounds (compounds 4, 5 and 6) were used for M.D.
simulation. The M.D. simulation was performed with the
YASARA Dynamics software version 18.4.24.W.64 (Krieger
et al., 2004). It was carried out throughout 25ns in an explicit
water environment, at constant pressure, using an AMBER14
force field periodic cell boundary condition. The model was
simulated at 295 K (25ꢀC), then the complex was equilibrated
for a 2ns period. After equilibration, a production M.D. was
run for 25ns at a constant temperature and pressure and
snapshots were saved at every 100ps toward M.D. trajectory
for more analysis (Islam et al., 2020). Root-mean-square devi-
ation (RMSD), root-mean-square fluctuation (RMSF), the
radius of gyration (Rg), solvent accessible surface area
(SASA), and the total number of H-bond count were calcu-
lated from the M.D. simulations (A. A. & E. B., 2020; Nisha
et al., 2020). The RMSD, RMSF, Rg, total number of hydrogen
bonds and the secondary structure fractions of protein in the
complex values were analyzed for each simulation. The RMSF
for the average equilibrium conformation is often used as an
indicator of protein rigidity, which is considered to be a fea-
ture related to the resistance to unfolding (Kuzmanic &
Zagrovic, 2010).
Figure 2. Optimized geometry of products 4 (a), 5 (b) and 6 (c) at B3LYP/6-
311 þ G(d,p) level.
2.6. Molecular docking
We have chosen the compounds 4, 5 and 6 to evaluate their
interaction toward an anti-apoptotic protein which is block-
ing cells death, inhibiting them will induce apoptosis and
combating cancer cells by killing them with apoptosis phe-
nomenon (Longley & Johnston, 2005; Montero & Letai, 2018).
They play a crucial role also in the cancer of beast, especially
in MCF-7 cancer as U. Brinkmann et al. published in one of
them work Effects of BCL-2 over expression on the sensitivity
of MCF-7 breast cancer cells (Brinkmann et al., 1997; Byadi et
al., 2020).
3. Results and discussion
The first step in synthesizing the desired hybrid
Thiazolidinone-Carvone ꢂ1,2,3-triazole 6 is the FeCl3 x 10H2O
catalyzed chlorination of natural (R)-carvone 1 using bleach
(NaOCl) as chlorine source. The resulting monochloro(R) car-
vone derivative 2 was then treated with sodium azide (NaN3)
under gentle reflux of acetone to lead to the azide car-
vone 3.
The crystal structure of Bcl-2 protein download from the
protein data bank website (http://www.rcsb.org), with a reso-
ꢁ
lution of 2.10 Å (PDB ID 4ieh) (Toure et al., 2013), we pre-
pared for docking with three ligands (4, 5 and 6), the ligands
€
also are prepared (Turkes¸ et al., 2020; Taslimi et al., 2020)
and optimized. During protein preparation, the water mole-
cules were removed along with the addition of hydrogen
atoms by satisfying the MMFF force fields implemented in carvone
The third step of our process to obtain the target product
6 was achieved by a [3 þ 2]cycloaddition reaction of azide
3
(used
as
1,3-dipole)
with
dimethyl

6
M. E. HACHIM ET AL.
Figure 3. HOMO, LUMO orbitals for compounds 4(a), 5(b) and 6 (c).
acetylenedicarboxylate (DMAD) (used as dipolarophile); the They were mainly characterized by the disappearance of
reaction was carried out in dry toluene under reflux for 3 h. the propenylic methyl signals and instead, the appear-
It is worth mentioning that compounds 2, 3 and 4 were
obtained in high yields of 75%, 79% and 69% respectively
(Scheme 1).
ance of methylenic resonances due to CH2Cl and CH2N3
groups for compound 2 and compound 3, respectively
(Table 1).
According to Scheme 2, product 4 was treated with thio-
semicarbazide in the presence of traces of sulphuric acid to
obtain the 1,2,3-triazole thiosemicarbazone 5 in 76% yield.
The final step of the 1,2,3-triazole-thiazole-thiazolidinone-6
synthesis was the heterocyclization of thiosemicarbazone 5
by the ethyl bromoacetate in the presence of sodium acetate
in ethanol under reflux conditions according to a procedure
described in the following paper (Bimoussa et al., 2019)
(Scheme 2).
As regards to the newly prepared 1,2,3-triazole 4, 1,2,3-tri-
azole-thiosemicarbazone 5 and 1,2,3-triazole-thiazolidinone 6,
they were fully characterized by their NMR (1H & 13C) and
HRMS spectral data.
Indeed, HRMS spectra of 4, 5 and 6, show the correspond-
ing pseudo molecular ion [M þ H]þ at m/z ¼ 334.1349 (4),
m/z ¼ 407.1452 (5), and m/z ¼ 447.1481 (6) corroborating
their molecular formula C16H19N3O5 (4), C17H22N6O4S (5), and
C19H22N6O5S (6).
In addition to the signals characterizing the methylene
and methylidene groups H2C3 (d1H: 4.3- 4.7; d 13C: 49-52)
and H2C4 (d1H: 4.7- 5.8; d 13C: 110-112), we notice in com-
pounds 4, 5 and 6 NMR spectra resonances attributable to
C4’ (d 13C: 132-135) and C5’ (d 13C: 122-125) carbons of the
1
3.1. HNMR and13C NMR spectrum analysis
It should be noted here that the spectral data of both
mono-chloro and azido-carvone derivatives 2 and 3 were
in perfect agreement with those described in the litera- triazole nucleus and its two methoxy substituents (d1H: 3.75-
ture (Weinges & Schwarz, 1993; Galstyan et al., 2018). 4.00; d 13C: 50-54) (Table 2).

JOURNAL OF BIOMOLECULAR STRUCTURE AND DYNAMICS
7
Scheme 3. The general mechanism for the synthesis of compound 6.
Table 3. Electronics properties and global reactivity indexes of reagents.
see Figure 2. The called highest occupied molecular orbital
(HOMO) and the lowest unoccupied molecular orbital
(LUMO) are the most important orbitals, which can define
the reactivity ability of a molecule (Rachedi et al., 2019; Uzun
et al., 2019). The LUMO orbital determines the ability of a
compound to accept or donatean electron in the case of
HOMO (Sakthivel et al., 2018; Hachim et al., 2019; Hachim et
al., 2020). HOMO and LUMO orbitals of 4, 5 and 6 were
examined and are shown in Figure 3. The analysis indicates
that the HOMO orbitals are located over the six-membered
ring in both compounds 4 and 5, while LUMO orbitals are
located over the triazole moieties. However, in the case of
compound 6, the HOMO and LUMO orbitals are located over
the substituted carvone ring. The analysis of the energy gap
(Egap) between HOMO energy and LUMO energy shows that
compounds 4 and 5 have relatively high value indicates that
these structures are low reactive compared to compound 6
(see Figure 3).
l(eV)
g (eV)
x (eV)
N (eV)
DMAD
3
P4
Thiosemicarbazide
P5
P6
ꢂ4.97
ꢂ4.32
ꢂ4.48
ꢂ3.18
ꢂ4.03
ꢂ4.05
6.47
5.22
4.99
6.10
4.11
4.18
1.91
1.79
2.01
0.83
1.98
1.96
0.92
2.19
2.14
2.89
3.03
2.98
The triazole derivative 5 is distinguished by the thiocarba-
1
mide moiety revealed in its H RMN spectrum as two singlets
at 6.16 and 9.62 ppm. These signals are absent in the 1H
RMN spectrum of the triazole-thiazolidinone hybrid 6 which
is further characterized by resonances of the methylene
group at C5" position of the thiazolidinone nucleus (d1H:
3.77; d 13C: 30.1). All the above mentioned spectroscopic
data of the 1,2,3-triazoles 4 and 5 and the triazole-thiazolidi-
none hybrid 6 corroborate the corresponding structures and
were in full accord with the literature (Ousidi et al., 2016;
Taia et al., 2020).
3.3. Mechanistic studies
In order to explain these experimental results and to find a
precise understanding of the reaction mechanism, calcula-
tions of the density functional theory (DFT) using the B3LYP/
6-311 þ G(d,p) level (Scheme 3) were performed.
3.2. Optimized molecular structure of compounds 4, 5
and 6
Recently, the analysis of polar reactions using global
reactivity indices has shown that these are powerful tools
The geometric structure of new carvone-1,2,3-triazole deriva-
tives was optimized using DFT at B3LYP/6-311Gþ(d,p)level, for understanding the reaction mechanism measured by

8
M. E. HACHIM ET AL.
Figure 4. DFT calculations on the mechanism for the synthesis of the compound 6 at B3LYP/6-311 þ G(d,p) level.
Figure 5. Optimized structures of all reported structures for the synthesis of compounds 4 and 5.

JOURNAL OF BIOMOLECULAR STRUCTURE AND DYNAMICS
9
the value of the global electron density transfer (GEDT)
value (Domingo et al., 2020). In this regard, the global
electrophilicity and nucleophilicity scales of reactants are
calculated and the results are given in Table 3. The value
of the electronic chemical potential of 3 (m¼ ꢂ4.32 eV) is
considerably higher than that of the DMAD. These results
indicate that along this cycloaddition reaction the global
electron density transfer (GEDT) should take place from 3
to DMAD. The 3 has a strong electrophile (Ȧ ¼ 1.79 eV)
and a moderate nucleophile (N ¼ 2.19 eV) within the elec-
The mechanistic study for the synthesis of compounds 4,
5 and 6 was performed using DFT calculation in the pres-
ence of the corresponding solvent and the results of all
steps are summarized in Figure 4. The first step is the [3 þ 2]
cycloaddition between compound 3 and dimethyl acetylene
dicarboxylate which conduct to the formation of compound
4 and the activation energy in this step is 30.41 kcal/mol.
Further, the compound 4 was reacted with thiosemicarba-
zide forming the compound 5 in two steps, the first one
is the formation of intermediate
1
(IM1) via TS2
ꢁ
trophilicity and nucleophilicity scales (Domingo & Perez,
(DG ¼ 60.13 kcal/mol) and the second one is endothermic by
50.33 kcal/mol.Finally, compound 6 was formed by the reac-
tion between the compound 5 and ethyl bromoacetate via
the formation of intermediate 2 (IM2), see Figure 4. The
optimized structure of TS1, TS2 and TS3 was illustrated in
Figure 5.
2011; Parr et al., 1999).On the other hand, the DMAD has
a
strong electrophile and marginal nucleophile. These
results confirm that along with this reaction the 3 acts as
a nucleophile and DMAD as anelectrophile. The analysis of
global reactivity indexes for both compound 4 and thiose-
micarbazide indicates that along this reaction GEDT should
take place from thiosemicarbazide (m¼ ꢂ3.18 eV) to com-
pound 4 (m ¼ ꢂ4.48 eV) and the compound thiosemicarba-
zide act as a nucleophile and 4 as an electrophile. The
analysis of the global reactivity indexes of compounds 4, 5
and 6 shows that the compounds 5 and 6 are more
nucleophile and less electrophile compared with com-
pound 4.
3.4. Cytotoxic activity evaluation of compounds 4, 5
and 6
Anti-proliferative activities of the 1,2,3-triazole 4 and their
derivatives 5 and 6 have been evaluated in three cancer cell
lines, including fibrosarcoma (HT-1080), lung carcinoma (A-
549) and breast adenocarcinoma (MCF-7) cell lines. Cells
were treated with different concentrations in a range of 6.25-
100 lM during 24 h, using Doxorubicin (DOX) as a positive
control, and cell growth has been quantified using the MTS
assay. The IC50 values are presented in Table 4.
Table 4. Evaluation of the cytotoxic effect of the synthesized compounds 4, 5
and 6 against human HT-1080, A-549 and MCF-7 cancer cells.
IC50 (mM)
Data in Table 4 show that structural modification of the
Compound
HT-1080
A-549
MCF-7
(R)-carvone
1
into 1,2,3-triazole-carvone
4
moderately
1
4
5
6
>100
45.64
29.25
55.73
5.38
>100
39.19
31.62
71.24
6.79
>100
51.37
26.02
66.25
6.12
increased the anti-proliferative activity against the three cell
lines, with IC50 values higher than 100 mM in the case of (R)-
carvone 1 and ranging from 39.19 to 51.37 lM in the case of
the derivative 4. Besides, the 1,2,3-triazole ring was kept, and
Doxorubicin
Figure 6. The 3 D interaction of compound 5 with the binding site of Bcl-2 protein.
Table 5. The results of docking with auto-dock using Yasara software.
Compounds
BE [kcal/mol]
DK [mM]
Contacting receptor residues
1
4
5
6
5.53
6.40
7.08
6.23
88.4
20.5
6.5
ALA 59 ASP62 PHE63 ARG66 TYR67 TRP103 GLY104 VAL107 PHE157 TYR161
ALA59 ASP62 PHE63 ARG66 TYR67 ASN102 TRP103 GLY104 ARG105 VAL107 ALA108 PHE157 TYR161
ALA59 ASP62 PHE63 ARG66 TYR67 ASN102 TRP103 GLY104 VAL107 PHE157 LEU160 TYR161
PHE 63 ASP 70 PHE 71 GLU 73 MET74 GLN77 VAL92 GLU95 LEU96 ALA108 PHE112
27.0

10
M. E. HACHIM ET AL.
we examined the effect of the thiosemicarbazone group. As (water, NaCl, etc.). Molecular dynamics (M.D.) simulation is one
shown in, Table 4, due to the addition of the carbamothioyl
group (-CS-NH2), the anti-proliferative activity the derivative
5 induced the highest level of cytotoxicity with IC50 values
between 26 and 31 lM. Finally, the derivative 6, after incorp-
oration of the thiazolidinone function, showed moderate
cytotoxic effects with IC50 values ranging from 55.73 to
66.25 mM. Altogether, these data show that only the addition
of the carbamothioyl group (-CS-NH2) to derivative 4 in order
to obtain derivative 5 leads to a higher cytotoxic effect.
In terms of Structure-Activity relationship, we can state
from a cytotoxic profile that, the antiproliferative activity of
these compounds could be associated with the combination
of dimethyl 1,2,3-triazole-4,5-dicarboxylate core and carba-
mothioyl group (-CS-NH 2 ). This lets us suggest that a carba-
mothioyl group attached to the terpene skeleton via
carbonyl group is essential for better efficiency in antiproli-
ferative activity while replacing this group by a thiazolidi-
none core, resulting in a loss of the activity. We believe that
such observation is associated with the absence of the differ-
ent interactions between the heteroatomic elements (sulfur
& nitrogen) of product 5 with the target. For this reason, a
more in-depth experimental study of the Structure-Activity
Relationship on product 5 would be necessary to better
understand the mechanism of this anti-cancer activity, par-
ticularly, on the two 1,2,3-triazole and carbamothioyl frag-
ments. Also, molecular modelling could be beneficial to
develop other analogues with improved activity.
between varieties of modern computational tools that are com-
monly used in protein modelling engineering. This area is gaining
tremendous interest as an affordable and effective alternative for
the rational or knowledge-based design of proteins.
To verify the stability of the systems by the molecular
dynamic approach, we have evaluated the mean value of the
total potential energy of the formed complex. The average
of the total potential energy found for the complexes 06-Bcl-
2, 05-Bcl-2 and 04-Bcl-2 were ꢂ392300, ꢂ399500 and
ꢂ382250 Kj/mol.
A standard physical parameter to identify conformational
variations of the structure compared to the original structure
during molecular dynamics is the RMSD (Root Mean Squared
Deviation). At the same time, the Root Mean Square
Fluctuation (RMSF) is used to identify the area of the protein
fluctuating during the simulation. The radius of gyration (Rg),
a parameter related to the tertiary structural volume of the
protein, is an indicator of the compactness of the protein
structure. The hydrogen bond number between the solvent
and the complex is also a reliable indicator of useful pro-
tein folding.
Analyzing the RMSD of the complexes protein-ligand is
shown in the Figure 7, it can be said that the average RMSD
of the complexes 6, 5 and 4 is about 2 Å, 1.8 Å and 2.1 Å
respectively, confirming the stability of the complexes and
the binding affinity between the ligands and the receptors is
very high. Thus, with the RMSD we have investigated the sta-
bility of the molecular structure of the protein compared to
its original conformation. To reveal the complexes, fluctua-
tions occurred during the M.D. time scale, especially for the
complex of compound 5 and Bcl-2 protein.
The plot of RMSF was analyzed in terms of time for the
complex and presented in Figure 8. The RMSF of complex
and each part of interaction is illustrated, confirming stable
complex, the flexibility of large amino acids is required,
including binding site’ amino acid. Such as GLU73, MET74
and LEU160, The RMSF of all-amino acid was 2 0.07, which
is considered as a standard error of RMSF, indicating that the
complexes flexibly were enough and normal.
The average Rg values for 04-Bcl-2 complexes, 05-Bcl-2
complexes and 06-Bcl-2 complexes are found in the average
of 14.7 Å, by following their stability in terms of flexibility of
fluctuations during the simulation, It should be noted that
the Gyration radius indicating good compactness of protein
must be less than 22.2 (Galzitskaya & Garbuzynskiy, 2008).
So, our complexes are stable. The protein has a good attrac-
tion to compounds 4,5 and 6 through its binding site (see
Figure 9).
3.5. Molecular docking
We classified the ligand-based on two criteria, binding energy
and dissociation constant. The interactions between ligands
and the hydrophobic side chains of proteins contribute signifi-
cantly to the binding free energy. The hydrophobic residues
mutually repel water and other polar groups and result in a net
attraction of the non-polar groups of the ligand. Binding affin-
ity is typically measured and announced by the equilibrium dis-
sociation constant (K.D.), which is used to examine and rank
order strengths of bi-molecular interactions. The smaller the
K.D. Value, the greater the binding affinity of the ligand for its
receptor. The binding interaction between the best com-
pounds, which has a good score, is presented in the binding
pocket of Bcl-2 protein in the following Figure 6.
Comparing the result of Table 5 (in silico) with Table 4
(experimental), it can be said that both the results in silico and
the experimental results are correlated: Compound 5 has the
highest anti-cancer activity, followed by compound 4 and com-
pounds 6. At the same time, compound 1 in silico nether was
not active in vitro against the Bcl-2 protein in cancer control. For
further analysis, we will use molecular dynamics to evaluate the
stability of each complex formed by ligands 4, 5 and 6.
We calculated the hydrogen bonds formed between the
complexes and water in the solvent environment within
0.35 nm during 25 ns M.D. simulations and we presented them
in Figure 10. We observed that the complexes 06-Bcl-2 com-
plex, 05-Bcl-2 complex, 04-Bcl-2 complex, Ref-Bcl-2 bind to the
solvent with 260, 267, 265 and 270 hydrogen bonds, respect-
ively, indicating the excellent stability of the complex 5 com-
3.6. Molecular dynamic
The molecular dynamics method is used to describe the complex
stability in its environment formed by the other constitution paring to the reference compound.

JOURNAL OF BIOMOLECULAR STRUCTURE AND DYNAMICS
11
Figure 7. Complex RMSD from the starting structure as a function of simulation time. a: 06-Bcl-2 complex, b: 05-Bcl-2 complex, c: 04-Bcl-2 complex, a-carbon
atom RMSD, the backbone RMSD and of all heavy atoms RMSD.
Figure 8. a-carbon atom RMSF values (Å) as a function of the residue number, during the entire simulation.
cytotoxic activity revealed a moderate to low anti-prolifera-
tive activity of the compounds against the three cancer cell
lines except for compound 5, which showed a higher cyto-
toxic effect. The global reactivity indexes explained the regio-
selectivity for the formation of compound 6, and the
4. Conclusion
New compounds from naturally (R)-carvone containing 1,2,3-
triazole were synthesized and characterized based on their
1H NMR and 13C NMR analysis. The evaluation of the

12
M. E. HACHIM ET AL.
Figure 9. The radius of gyration of the solute as a function of simulation time, a: 06-Bcl-2 complex, b: 05-Bcl-2 complex, c: 04-Bcl-2 complex, d: Ref-Bcl-2 complex.
Figure 10. Number of hydrogen bonds in the solute as a function of simulation time, a: 06-Bcl-2 complex, b: 05-Bcl-2 complex, c: 04-Bcl-2 complex, d: Ref-Bcl-
2 complex.
theoretical results are in agreement with experimental find-
ings. Besides, we estimated the binding affinity of our com-
pounds to Bcl-2, an essential protein that is one of the main
targets in cancer research. Molecular docking and subse-
quent studies on molecular dynamics have shown the higher
stability of the complex between Bcl-2 and compound 5
compared to other compounds. Therefore, we propose com-
pound 5 for further in vitro evaluation.
Acknowledgements
We are grateful to Cadi Ayyad University and Chouaïb Doukkali
University for providing financial support.
Disclosure statement
No potential conflict of interest was reported by the author(s).

JOURNAL OF BIOMOLECULAR STRUCTURE AND DYNAMICS
13
properties and interaction with bovine serum albumin. Molecules,
23(11), 2991. https://doi.org/10.3390/molecules23112991
ORCID
Mouhi Eddine Hachim
http://orcid.org/0000-0001-7692-5323
Galzitskaya, O. V.,
& Garbuzynskiy, S. O. (2008). Molecular Biology,
Said Byadi
http://orcid.org/0000-0003-0913-2897
144–154.
Lahoucine Bahsis
Crtomir Podlipnik
Aziz Auhmani
My Youssef Ait Itto
http://orcid.org/0000-0002-7252-6680
http://orcid.org/0000-0002-8429-0273
http://orcid.org/0000-0002-0459-6935
http://orcid.org/0000-0003-1876-7544
Geerlings, P., De Proft, F., & Langenaeker, W. (2003). Conceptual density
functional theory. Chemical Reviews, 103(5), 1793–1874. https://doi.
org/10.1021/cr990029p
Hachim, M. E., Sadik, K., Byadi, S., & Aboulmouhajir, A. (2020). Electronic
investigation and spectroscopic analysis using DFT with the long-
range dispersion correction on the six lowest conformers of 2.2.3-tri-
methyl pentane. Journal of Molecular Modeling, 26(7). https://doi.org/
10.1007/s00894-020-04430-4
Hachim, M. E., Sadik, K., Byadi, S., Van Alsenoy, C., & Aboulmouhajir, A.
(2019). Ab initio study on the six lowest energy conformers of iso-
octane: conformational stability, barriers to internal rotation, natural
bond orbital and first-order hyperpolarizability analyses, UV and NMR
predictions, spectral temperature sensitivity, and scaled vibrational
assignment. Journal of Molecular Modeling, 25(8). https://doi.org/10.
1007/s00894-019-4105-5
Islam, R., Parves, R., Paul, A. S., Uddin, N., Rahman, S., Al Mamun, A.,
Hossain, N., Ali, A., & Halim, M. A. (2020). Journal of Biomolecular
Structure and Dynamics, 0, 1–12.
ꢀ
References
A. A., E. & E. B., A. (2020). Journal of Biomolecular Structure and Dynamics.
Bimoussa, A., Koumya, Y., Abouelfida, A., Itto, M. Y. A., Benyaich, A.,
Mentre, O., Ketatni, E. M., Auhmani, A.,
& Auhmani, A. (2019).
Hemisynthesis, crystal structure and inhibitory effect of sesquiterpenic
thiosemicarbazones and thiazolidin-4-ones on the corrosion behaviour
of stainless steel in 1 M H2SO4 solution. Acta Crystallographica.
Section C, Structural Chemistry, 75(Pt 6), 623–632. https://doi.org/10.
1107/S2053229619005631
Brahmkshatriya, P. P., & Brahmkshatriya, P. S. (2013). Natural products:
Phytochemistry, botany and metabolism of alkaloids, phenolics and ter-
penes (pp. 2665–2691) (K. G. Ramawat & J.-M. Merillon, Eds.). Springer.
ꢁ
€
Kalaycı, M., Turkes¸, C., Arslan, M., Demir, Y., & Beydemir, S¸. (2021). Novel
Brinkmann, U., Mansfield, E., & Pastan, I. (1997). Effects of BCL-2 overex-
pression on the sensitivity of MCF-7 breast cancer cells to ricin, diph-
theria and Pseudomonas toxin and immunotoxins. Apoptosis : An
International Journal on Programmed Cell Death, 2(2), 192–198. https://
doi.org/10.1023/A:1026468532413
benzoic acid derivatives: Synthesis and biological evaluation as multi-
target acetylcholinesterase and carbonic anhydrase inhibitors. Archiv
Der Pharmazie, 354(3), 2000282. https://doi.org/10.1002/ardp.
202000282
€
Kilic, A., Beyazsakal, L., Is¸ı k, M., Turkes¸, C., Necip, A., Takı m, K., &
ꢀ
Beydemir, S¸. (2020). Mannich reaction derived novel boron com-
plexes with amine-bis(phenolate) ligands: Synthesis, spectroscopy and
in vitro/in silico biological studies. Journal of Organometallic
Chemistry, 927, 121542. https://doi.org/10.1016/j.jorganchem.2020.
121542
Byadi, S., Eddine, H. M., Sadik, K., Podlipnik, C., & Aboulmouhajir, A.
Fingerprint-based 2D-QSAR models for predicting Bcl-2 inhibitors
affinity. Letters in Drug Design & Discovery, 17(10), 1206–1215. https://
doi.org/10.2174/1570180817999200414155403
Crowell, P. L. (1999). Prevention and therapy of cancer by dietary mono-
terpenes. The Journal of Nutrition, 129(3), 775S–778S. https://doi.org/
10.1093/jn/129.3.775S
Krieger, E., Darden, T., Nabuurs, S. B., Finkelstein, A., & Vriend, G. (2004),
683, 678–683. https://doi.org/10.1002/prot.20251
Kuzmanic, A., & Zagrovic, B. (2010). Determination of ensemble-average
pairwise root mean-square deviation from experimental B-factors.
Biophysical Journal, 98(5), 861–871. https://doi.org/10.1016/j.bpj.2009.
11.011
Longley, D. B., & Johnston, P. G. (2005). Molecular mechanisms of drug
resistance. The Journal of Pathology, 205(2), 275–292. https://doi.org/
10.1002/path.1706
Domingo, L. R. (2014). A new C–C bond formation model based on the
quantum chemical topology of electron density. RSC Advances, 4(61),
32415–32428. https://doi.org/10.1039/C4RA04280H
ꢁ
Domingo, L. R., & Perez, P. (2011). The nucleophilicity N index in organic
chemistry. Organic Biomolecular Chemistry, 9(20), 7168–7175.
https://doi.org/10.1039/c1ob05856h
&
ꢁ
ꢁ
ꢁ
Domingo, L. R., Rıos-Gutierrez, M., & Perez, P. (2016). Molecules. https://
Mansouri, A.-E. E., Maatallah, M., Benhassou, H. A., Moumen, A., Mehdi,
A., Snoeck, R., Andrei, G., Zahouily, M., & Lazrek, H. B. (2020). Design,
synthesis, chemical characterization, biological evaluation, and dock-
ing study of new 1,3,4-oxadiazole homonucleoside analogs.
Nucleosides, Nucleotides & Nucleic Acids, 39(8), 1088–1107. https://doi.
org/10.1080/15257770.2020.1761982
Mansouri, A.-E. E., Oubella, A., Maatallah, M., AitItto, M. Y., Zahouily, M.,
Morjani, H., & Lazrek, H. B. (2020). Design, synthesis, biological evalu-
ation and molecular docking of new uracil analogs-1,2,4-oxadiazole
doi.org/10.3390/molecules21060748
ꢁ
ꢁ
Domingo, L., Rıos-Gutierrez, M., & Soriano, J. C. (2020). Understanding
the origin of the regioselectivity in non-polar [3þ 2] cycloaddition
reactions through the molecular electron density theory. Journal of
Physical Organic Chemistry, e4062, 1–11. https://doi.org/10.1002/poc.
4062.
Feddouli, A., Itto, M. Y. A., Ali, M. A., Hasnaoui, A., & Riahi, A. (2006).
Efficient approach for the synthesis of novel functionalized isoxazo-
lines from limonene. Synthetic Communications, 36(23), 3617–3624.
https://doi.org/10.1080/00397910600943709
hybrids as potential anticancer agents. Bioorganic
& Medicinal
Chemistry Letters, 30(19), 127438. https://doi.org/10.1016/j.bmcl.2020.
127438
ꢂ
Feddouli, A., Itto, M. Y. A., Hasnaoui, A., Villemin, D., Jaffres, P.-A., Santos,
J. S.-D. O., Riahi, A., Huet, F., & Daran, J.-C. (2004). One pot diastereo-
selective synthesis of new chiral spiro-1,3,4-thiadiazoles and 1,4,2-oxa-
thiazoles from (1 R )-thiocamphor. Journal of Heterocyclic Chemistry,
41(5), 731–735. https://doi.org/10.1002/jhet.5570410513
Fei, J., Zhou, L., Liu, T., & Tang, X.-Y. (2013).Pharmacophore modeling, vir-
tual screening, and molecular docking studies for discovery of novel
Akt2 inhibitors. International Journal of Medical Sciences, 10(3),
265–275. https://doi.org/10.7150/ijms.5344
Frisch, M. J., Trucks, G. W., Schlegel, H. B., Scuseria, G. E., Robb, M. A.,
Cheeseman, J. R., Scalmani, G., Barone, V., Petersson, G. A., Nakatsuji,
H., Li, X., Caricato, M., Marenich, A. V., Bloino, J., Janesko, B. G.,
Gomperts, R., Mennucci, B., Hratchian, H. P., … ..Fox, D. J.. (2009).
Gaussian. 09. Revision D.01, Gaussian Inc.
Galstyan, A. S., Martiryan, A. I., Grigoryan, K. R., Ghazaryan, A. G.,
Samvelyan, M. A., Ghochikyan, T. V., & Nenajdenko, V. G. (2018).
Synthesis of Carvone-Derived 1,2,3-triazoles study of their antioxidant
Mansouri, A.-E. E., Zahouily, M., & Lazrek, H. B. (2019). HMDS/KI a simple,
a cheap and efficient catalyst for the one-pot synthesis of N -func-
tionalized pyrimidines. Synthetic Communications, 49(14), 1802–1812.
https://doi.org/10.1080/00397911.2019.1602655
Moitessier, N., Englebienne, P., Lee, D., Lawandi, J., & Corbeil, C. R.
(2008). Towards the development of universal, fast and highly accur-
ate docking/scoring methods: a long way to go. British Journal of
Pharmacology, 153(S1), S7–S26. https://doi.org/10.1038/sj.bjp.0707515
Montero, J., & Letai, A. (2018). Why do BCL-2 inhibitors work and where
should we use them in the clinic? Cell Death and Differentiation, 25(1),
56–64. https://doi.org/10.1038/cdd.2017.183
Newman, D. J., Cragg, G. M., & Snader, K. M. (2000). The influence of nat-
ural products upon drug discovery. Natural Product Reports, 17(3),
215–234. https://doi.org/10.1039/a902202c
Nisha, M., Sakthivel, R., Velmurugan, D., & Michael, G. M. (2020). Journal
of Biomolecular Structure and Dynamics.

14
M. E. HACHIM ET AL.
ꢁ
Oubella, A., Ait Itto, M. Y., Auhmani, A., Riahi, A., Daran, J.-C., & Auhmani, Taia, A., Essaber, M., Oubella, A., Aatif, A., Bodiguel, J., Jamart-Gregoire,
A. (2020). Crystal structure of (R)-5-[(R)-3-(4-chloro-phen-yl)-5-
methyl-4,5-di-hydro-isoxazol-5-yl]-2-methyl-cyclo-hex-2-enone. Acta
Crystallographica. Section E, Crystallographic Communications, 76(Pt 3),
400–403. https://doi.org/10.1107/S2056989020001991
Oubella, A., Ait Itto, M. Y., Auhmani, A., Riahi, A., Robert, A., Daran, J.-C.,
Morjani, H., Parish, C. A., & Esseffar, M. (2019). Diastereoselective syn- Taslimi, P., Is¸ı k, M., Turkan, F., Durgun, M., Turkes¸, C., Gulc¸in, I., &
B., Itto, M. Y. A., & Morjani, H. (2020). Synthesis, characterization, and
biological evaluation of new heterocyclic systems 1, 2, 3-triazole-iso-
xazoline from eugenol by the mixed condensation reactions. Synthetic
Communications, 50(13), 2052–2065. https://doi.org/10.1080/00397911.
2020.1762224
_
€
€
€
thesis and cytotoxic evaluation of new isoxazoles and pyrazoles with
monoterpenic skeleton. Journal of Molecular Structure, 1198, 126924.
https://doi.org/10.1016/j.molstruc.2019.126924
Beydemir, S¸. (2020). Benzenesulfonamide derivatives as potent acetyl-
cholinesterase, a-glycosidase, and glutathione S-transferase inhibitors:
biological evaluation and molecular docking studies. Journal of
Biomolecular Structure, 1–12.
Oubella, A., Fawzi, M., Auhmani, A., Riahi, A., Morjani, H., Robert, A., &
ꢁ
ꢁ
Itto, M. Y. A. (2020). Synthesis and antitumor activity of novel hetero- Toure, B. B., Miller-Moslin, K., Yusuff, N., Perez, L., Dore, M., Joud, C.,
cyclic systems with monoterpenic skeleton combining dichlorocyclo-
propane and 1,3,4-thiadiazole nucleus. ChemistrySelect, 5(21),
6403–6406. https://doi.org/10.1002/slct.202001284
Michael, W., Dipietro, L., Van Der Plas, S., Mcewan, M., Lenoir, F., Hoe,
M., Karki, R., Springer, C., Sullivan, J., Levine, K., Fiorilla, C., Xie, X.,
Kulathila, R.,
… Visser, M. (2013). The Role of the acidity of N-
Ousidi, A. N., Itto, M. Y. A., Auhmani, A., Riahi, A., Ayouchia, H. B. E.,
Anane, H., & Aouad, N. E. (2016). 10.
Heteroaryl sulfonamides as inhibitors of Bcl-2 family protein–protein
interactions. ACS Medicinal Chemistry Letters, 4, 2, 186-190. https://doi.
org/10.1021/ml300321d
ꢁ
Parr, R. G., Szentpaly, L. v., & Liu, S. (1999). Electrophilicity index. Journal
€
of the American Chemical Society, 121(9), 1922–1924. https://doi.org/ Turkes¸, C. (2019). Investigation of potential paraoxonase-I inhibitors by
10.1021/ja983494x
Rachedi, K. O., Ouk, T.-S., Bahadi, R., Bouzina, A., Djouad, S.-E., Bechlem,
K., Zerrouki, R., Ben Hadda, T., Almalki, F., & Berredjem, M. (2019).
kinetic and molecular docking studies: Chemotherapeutic drugs.
Protein & Peptide Letters, 26(6), 392–402. https://doi.org/10.2174/
0929866526666190226162225
€
Synthesis, DFT and POM analyses of cytotoxicity activity of a-amido- Turkes¸, C., Beydemir, S¸., & Demir, Y. (2020). Some calcium-channel block-
phosphonates derivatives: Identification of potential antiviral O,O-
pharmacophore site. Journal of Molecular Structure, 1197, 196–203.
https://doi.org/10.1016/j.molstruc.2019.07.053
ers: Kinetic and in silico studies on paraoxonase-I. Journal of
Biomolecular Structure, 1–9.
€
€
€ €
Turkes¸, Cuneyt, Demir, Yeliz., & Beydemir, S¸ukru. (2020). Calcium chan-
nel blockers: Molecular docking and inhibition studies on carbonic
anhydrase I and II isoenzymes. Journal of Biomolecular Structure, 1–9.
Uzun, S., Esen, Z., Koc¸, E., Usta, N. C., & Ceylan, M. (2019). Experimental
and density functional theory (MEP, FMO, NLO, Fukui functions) and
antibacterial activity studies on 2-amino-4- (4-nitrophenyl) -5,6-dihy-
drobenzo [h] quinoline-3-carbonitrile. Journal of Molecular Structure,
1178, 450–457. https://doi.org/10.1016/j.molstruc.2018.10.001
ꢁ
ꢁ
Rıos-Gutierrez, M., Domingo, L. R., Esseffar, M., Oubella, A., & Ait Itto,
M. Y. (2020). Molecules. https://doi.org/10.3390/molecules25051085
Sakthivel, S., Alagesan, T., Muthu, S., Abraham, C. S., & Geetha, E. (2018).
Quantum mechanical, spectroscopic study (FT-IR and FT - Raman), NBO
analysis, HOMO-LUMO, first order hyperpolarizability and docking stud-
ies of a non-steroidal anti-inflammatory compound. Journal of Molecular
Structure, 1156, 645–656. https://doi.org/10.1016/j.molstruc.2017.12.024
€
Sever, B., Turkes¸, C., Altıntop, M. D., Demir, Y., & Beydemir, S¸. (2020). Weinges, K., & Schwarz, G. (1993). Radikalische Cyclisierung von Dienen,
Thiazolyl-pyrazoline derivatives: In vitro and in silico evaluation as
potential acetylcholinesterase and carbonic anhydrase inhibitors.
International Journal of Biological Macromolecules, 163, 1970–1988.
https://doi.org/10.1016/j.ijbiomac.2020.09.043
VI. Substratkontrollierte asymmetrische Synthese von (3aS,6aR)-
(þ)-3,3a,6,6a-Tetrahydro-2H-cyclopenta[b]furan-2-on. Liebigs Annalen
der
Chemie,
1993(7),
811–814.
https://doi.org/10.1002/jlac.
199319930149
ꢁ
Singh, G., Sachdeva, R., Rai, B., & Saini, G. S. S. (2017). Structure and
vibrational spectroscopic study of alpha-tocopherol. Journal of
Molecular Structure, 1144, 347–354. https://doi.org/10.1016/j.molstruc.
2017.05.037
Zimmermann, G. R., Lehar, J., & Keith, C. T. (2007). Multi-target therapeu-
tics: When the whole is greater than the sum of the parts. Drug
Discovery Today, 12(1-2), 34–42. https://doi.org/10.1016/j.drudis.2006.
11.008