Synthesis, crystal structure, spectroscopic characterization, Hirshfeld surface analysis, molecular docking studies and DFT calculations, and antioxidant activity of 2-oxo-1,2-dihydroquinoline-4-carboxylate derivatives

Article abstract of DOI:10.1016/j.molstruc.2019.03.103

A series of hydroquinolines derivatives (2a-2c) have been achieved by a cyclocondensation reaction. The synthesis of 2-oxo ?1,2-dihydroquinoline-4-carboxylic acid derivatives (3a-3c) has been carried out using alkylation reactions under phase transfer catalysis (PTC) conditions. The prepared products were characterized through spectroscopic NMR 1H and 13C, IR and single crystal X-ray diffraction techniques. 2D and 3D Hirshfeld surfaces (for 3a and 3b) studies were realized to understand non-bonding intermolecular interactions in solid phase crystal packing of the compound. With the optimized structures, the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) energies and clouds were obtained and evaluated. Good agreement was found between the calculated results and experimental data. Furthermore, the molecular docking study is performed to investigate binding patterns of the synthesized molecules (3a and 3b) into 1M17 inhibitor using Auto-Dock Vina program. The antioxidant activity of compounds (3a-3c) has been evaluated using DPPH free radical scavenging, ferric reducing (FRAP) power and β-carotene kinetic blanching assays. Both DPPH and FRAP methods confirmed that 3b had the best antioxidant activity followed by 3c. On the other hand, β-carotene bleaching assay showed the high activity of the product 3a.

Full text of DOI:10.1016/j.molstruc.2019.03.103

Accepted Manuscript
Synthesis, crystal structure, spectroscopic characterization, Hirshfeld surface
analysis, molecular docking studies and DFT calculations, and antioxidant activity of
2-oxo-1,2-dihydroquinoline-4-carboxylate derivatives
Yassir Filali Baba, Yusuf Sert, Youssef Kandri Rodi, Sonia Hayani, Joel T. Mague,
Damien Prim, Jerome Marrot, Fouad Ouazzani Chahdi, Nada Kheira Sebbar, El
Mokhtar Essassi
PII:
S0022-2860(19)30389-8
DOI:
https://doi.org/10.1016/j.molstruc.2019.03.103
MOLSTR 26372
Reference:
To appear in: Journal of Molecular Structure
Received Date: 24 December 2018
Revised Date: 16 March 2019
Accepted Date: 31 March 2019
Please cite this article as: Y. Filali Baba, Y. Sert, Y. Kandri Rodi, S. Hayani, J.T. Mague, D. Prim, J.
Marrot, F. Ouazzani Chahdi, N.K. Sebbar, E.M. Essassi, Synthesis, crystal structure, spectroscopic
characterization, Hirshfeld surface analysis, molecular docking studies and DFT calculations, and
antioxidant activity of 2-oxo-1,2-dihydroquinoline-4-carboxylate derivatives, Journal of Molecular
Structure (2019), doi: https://doi.org/10.1016/j.molstruc.2019.03.103.
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ACCEPTED MANUSCRIPT
The 3D Hirshfeld surface compounds 3a (up left) and 3b (up
right) and molecular structure 3a (bottom left) and 3b (bottom
right) of
2
-oxo-1,2-dihydroquinoline-4-carboxylate derivatives.

ACCEPTED MANUSCRIPT
Synthesis, crystal structure, spectroscopic characterization, Hirshfeld surface
analysis, molecular docking studies and DFT calculations, and antioxidant
activity of 2-oxo-1,2-dihydroquinoline-4-carboxylate derivatives.
a
b
a
a
c
Yassir Filali Baba , Yusuf Sert , Youssef Kandri Rodi , Sonia Hayani , Joel T. Mague , Damien
d
d
a
e,f*
f,g
Prim , Jerome Marrot , Fouad Ouazzani Chahdi , Nada Kheira Sebbar , El Mokhtar Essassi
a
Laboratoire de Chimie Organique Appliquée, Université Sidi Mohamed Ben Abdallah, Faculté des
Sciences et Techniques, BP 2202 Fez, Morocco.
b
Bozok University Department of Physics Yozgat/TURKEY
Department of Chemistry, Tulane University, New Orleans, LA 70118, USA
Lavoisier Institute, University of Versailles, 45 Avenue of the United States, France
Laboratoire de Chimie Appliquée et Environnement, Equipe de Chimie Bioorganique Appliquée, Faculté
c
d
e
des sciences, Université Ibn Zohr, Agadir, Morocco.
f
Laboratoire de Chimie Organique Hétérocyclique, Centre de Recherche des Sciences des Médicaments,
Pôle de Compétences Pharmacochimie, Mohammed V University in Rabat, Faculté des Sciences, Av. Ibn
Battouta, BP 1014 Rabat, Morocco.
g
Moroccan Foundation for Advanced Science, Innovation and Research (MASCIR), Rabat, Morocco.
_
________________
Corresponding authors:
Dr. Nada Kheira Sebbar
Tel: +212615011615; E-mail: snounousebbar@gmail.com

ACCEPTED MANUSCRIPT
Abstract
A series of hydroquinolines derivatives (2a-2c) have been achieved by a cyclocondensation
reaction. The synthesis of 2-oxo -1,2-dihydroquinoline-4-carboxylic acid derivatives (3a-3c) has
been carried out using alkylation reactions under phase transfer catalysis (PTC) conditions. The
prepared products were characterized through spectroscopic NMR 1H and 13C, IR and single
crystal X-ray diffraction techniques. 2D and 3D Hirshfeld surfaces (for 3a and 3b) studies were
realized to understand non-bonding intermolecular interactions in solid phase crystal packing of
the compound. With the optimized structures, the highest occupied molecular orbital (HOMO)
and the lowest unoccupied molecular orbital (LUMO) energies and clouds were obtained and
evaluated. Good agreement was found between the calculated results and experimental data.
Furthermore, the molecular docking study is performed to investigate binding patterns of the
synthesized molecules (3a and 3b) into 1M17 inhibitor using Auto-Dock Vina program. The
antioxidant activity of compounds (3a-3c) has been evaluated using DPPH free radical
scavenging, ferric reducing (FRAP) power and β-carotene kinetic blanching assays. Both DPPH
and FRAP methods confirmed that 3b had the best antioxidant activity followed by 3c. On the
other hand, β-carotene bleaching assay showed the high activity of the product 3a.
Keywords: Quinoline, X-ray, Hirshfeld surface, Molecular docking, DFT,
Antioxidant Activity
.

ACCEPTED MANUSCRIPT
1
. Introduction
The development of new chemically synthesized molecules represents a major strategy
for the discovery and preparation of new substances with biological interest [1]. Quinoline
heterocycles are a highly attractive ring system in organic chemistry, which are developing
rapidly as well as its synthetic versatility [2]. Molecules with quinoline motif are widespread
in natural products and drugs, due to their significant pharmacological properties. Quinoline
moiety is considered as an important pharmacophore and a privileged structure in medicinal
and industrial chemistry [3-6]. The increasing interest in the conception and synthesis of news
quinoline skeleton can be attributed also to their diverse therapeutic activities including
antimalarial [7], antibacterial [8], antifungal [9], anticancer [10] and anti-inflammatory [11-
1
2]. Some of these compounds are endowed with various activities, such as antituberculosis,
[
13] analgesic [14], cardiovascular [15], antibiotic [16], antihypertensive [17], tyrosine kinase
PDGF-RTK) inhibition [18], and anti-HIV [19-20]. Additionally, quinoline ring plays an
(
important role in the development of new antitumorals agents, as their derivatives have shown
excellent results through different mechanisms of action. They are growth inhibitors by cell
cycle arrest, apoptosis, angiogenesis inhibators, disruption of cell migration, and modulation
of nuclear receptor responsiveness [21-22]. Quinoline derivatives also find application in
agrochemicals and affect chemicals such as dyestuffs and corrosion inhibitors [23].
Due to their importance as substructures in a wide range of natural and designed
products, still great efforts continue to be directed to the development of new quinoline-based
structures. The work presented herein is devoted to the synthesize quinoline derivatives (2a-
2
c) using condensation in acetic acid of isatin with malonic acid in the presence of and
sodium acetate. Then, the compounds (2a-2c) obtained were alkylated using ethyl iodide in
the presence of K CO in DMF and tetra n-butyl ammonium bromide (TBAB) as catalyst. The
2
3
compounds (3a-3c) were obtained with good yields and their structures were elucidated on
the basis of spectral data (Fourier Transformer-Infrared (FT-IR) and Nuclear Magnetic
1
13
Resonance (NMR H and C) and confirmed in the cases of compounds 3a, 3b. by X-Ray
crystallographic studies The structures were also characterized by predicting the
corresponding spectroscopic data and Z-matrix coordinates using DFT calculations at the
B3LYP/6-31G (d,p) and molecular docking studies. The compounds 3a-3c are identified
through Hirshfeld surface analysis, molecular docking studies and DFT calculations.
Hirshfeld surfaces are used to investigate intermolecular interactions in crystal packing of the
compounds. The molecular docking study is performed with AutoDockVina program to
determine the binding modes at different poses in macromolecule of 3a and 3b into 1M17

ACCEPTED MANUSCRIPT
inhibitor. As well, molecular docking analysis are generally used to explore interactions
between receptor and ligand. Nowadays, the objective of several news researches is to
elaborate new molecules with biological activities. Besides, quinoline molecules are active
and have several biological activities [24-26], including antioxidant activity [27].
The free radicals are molecules or atoms, which include an unpaired electron at the level
of their external orbit. They take part in the appearance of many illness [28] by attacking the
cellular components. Synthetic antioxidants such as BHT (butylated hydroxytoluene), HA
(butylated hydroxy-anisole), TBHQ (tert-butyl- hydroquinone), BHA (butylated
hydroxyanisole), propyl gallate (PG) and ethoxyquin are suspected as being carcinogenic
substances [29]. Therefore, we started to evaluate the antioxidant activity of our compounds,
using three different methods free radical scavenging, ferric reducing power and β-carotene
bleaching.
2
2
. Material and methods
.1. Spectral data measurements
The spectroscopic characterization of the synthesized compounds is achieved by
recording NMR spectra (Bruker Avance DPX300) and FT-IR spectra in KBr pellets (JASCO
FTIR-4160 spectrometer). TLC and column chromatography were performed using silica
plates and silica gel, respectively. Melting points of compounds 3a-3c were determined in
open capillaries
.
2
.2. Preparation of compounds 3a-3c by alkylation reaction under PTC conditions.
The synthesis of 2-oxo-1,2-dihydroquinoline-4-carboxylic acid derivatives 3a-3c has
been carried out using alkylation reactions under phase transfer catalysis (CTP) conditions.
The action of ethyl iodide on compounds 2a-2c in DMF with heating for 6h in the presence of
tetra n-butylammonium bromide (BTBA) as a catalyst and K CO as a base, led to the
2
3
dialkylated products 3a-3c with good yields (Scheme 1).
O
OH
O
O
O
OH
OH
X
O
X
X
I
CH CH
2 3
CH COONa
O
3
+
O
N
acetic acid
reflux, 24 h
K CO ,BTBA
H
N
O
2
3
N
O
H
DMF
1
1
1
a :X= H
b :X= Cl
c :X= Br
3a :X= H
3b :X= Cl
3c :X= Br
2
2
2
a :X= H
b :X= Cl
c :X= Br
Scheme 1. Synthesis of ethyl-2-oxo-1,2-dihydroquinoline-4-carboxylate derivatives: 3a-3c.

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The structures of the synthesized compounds were determined by the usual spectroscopic
1
13
methods H NMR, C NMR, infrared and confirmed by X-Ray crystallographic studies in the
cases of compounds 3a and 3b.
2
*
.2.1. Experimental part:
General procedure synthesis of the compounds 2a-2c:
To a solution of 10 mmol of isatin and 10 mmol of malonic acid in 30 ml of acetic acid
was added 1 mmol of sodium acetate. The mixture was refluxed for 24 hours. After cooling,
00 ml of ice water are added. The precipitate obtained was washed several times with
ethanol [30].
1
2
-oxo-1,2-dihydroquinoline-4- carboxylic acid : 2a.
1
Yield (%) = 90; mp =553K; H NMR (300 MHz, CDCl ): 6.86 (s, 1H, CH); 7.2-8.16 (m,
3
1
3
4
H, CHarom) ; 12.17(s, 1H, NH ) ; 13.9 (s, 1 H, OH). CNMR (75 MHz, CDCl ): 167.2 (
3
COOH), 163. (C=O)), 141.7-139.8 (Cq-Cq), 131.3 (CHarom), 126.5 (CHarom), 123.8 (CH),
22.6 (Cq), 116.2 (CHarom).
1
-6-chloro-2-oxo-1,2-dihydroquinoline-4-carboxylic acid : 2b.
1
Yield (%) = 83; mp =553K; H NMR (300 MHz, CDCl ): 6.91 (s, 1H,CH) ; 7.32 (d, 1H,
3
3
1
JH-H = 9 Hz, (CH arom); 7.72 (d, 1H, 3JH-H = 9 Hz, CHarom); 8.22 (d, 1H, 4JH-H =
1
3
.8Hz, CH); 12.22(s, 1H, NH); 14.05 (s, 1 H, OH). C NMR (75 MHz, CDCl ): 166.69
3
(
(
-
COOH), 161.24 (C=O), 139.64-138.7 (Cq, Cq), 131.14 (CHarom), 126.7 (CHarom), 126.07
CH), 125.77 (CHarom), 118.08 (Cq), 117.42 (CHarom).
6-bromo-2-oxo-1,2-dihydroquinoline-4-carboxylic acid : 2c.
1
Yield (%) = 85; mp =553K; H NMR (300 MHz, CDCl ):6.98 (s, 1H,CH) ; 7.30 (d, 1H,
3
3
1
JH-H = 9 Hz, (CHarom); 7.7 (dd, 1H, 3JH-H = 9 Hz, 4JH-H = 1.8 Hz (CHarom); 8.42 (d,
1
3
H, 4JH-H = 1.8Hz, (CHarom); 12.22(s, 1H, NH) ; 14.05 (s, 1 H, OH). C NMR (75 MHz,
CDCl ): 166.7 ( COOH), 161.2 (C=O), 139.6-139 (Cq, Cq), 133.8 (CHarom), 128.7
3
(CHarom), 125.9 (CH), 118.3 (CHarom), 117.9 (Cq). 114.5 (CHarom).
*
General procedure synthesis of compounds 3a-3c:
-3
-3
To a solution of 10 mol of compound 2a-2c in 10 ml of DMF were added 2.5 × 10
-3
-3
mol of alkylating agent (ethyl iodide :4.10 mol of K CO and 0.01×10 mol of tetra n-
2
3
butylammonium bromide (TBAB). The reaction mixture was stirred at room temperature in
DMF for 6 h. [31-32] After removal of the salts by filtration, the solvent was evaporated
under reduced pressure and the residue obtained is dissolved in dichloromethane The organic
phase is dried over Na SO then concentrated under vacuum The compound obtained was
2
4
purified by chromatography on a column of silica gel (eluent: ethyl acetate / hexane (1/3)).

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Ethyl 1-ethyl-2-oxo-1,2-dihydroquinoline-4 -carboxylate: 3a.
1
Yield (%) = 86; Physical state: Purplecrystals ; mp =359 K; H NMR (300 MHz,
3
3
3
CDCl ): 1.39 (t, 3H, J = 7.2Hz, CH ), 1.42 (t, 3H, J = 6.9 Hz, CH ), 4.40 (q, 2H, J
H-H
3
H-H
3
H-H
3
3
=
6.9 Hz, CH ), 4.46 (q, 2H, J
= 7.2 Hz, CH ), 7.20 (s, 1H, CH), 7.30 (m, 1H, CHarom),
2
H-H
2
3
4
7
.45 (d, 1H, J
= 8.1 Hz, CHarom); 7.62 (m, 1H, CHarom); 8.35 (d, 1H, JH-H = 1.5 Hz,
H-H
1
3
CHarom). CNMR (75 MHz, CDCl ):165.46 (C=O), 160.95 (C=O), 139.36-138.90 (Cq-Cq),
3
1
31.07 (CHarom), 127.37 (CHarom), 124.20 (CHarom) 122.45 (CH), 117.80 (Cq), 114.35
-
1
(
CHarom), 61.99 (CH ), 37.64 (CH ), 14.38 (CH ), 12.63 (CH ). IR ( ν max, cm ): νC -
2 2 3 3 SP2
-1
-1
-1
-1
H=2979.33cm ;νC -H=2925.69cm ; νC=O=1717.63cm ; νC=C=1570.73-1511.83cm ;
SP3
Ethyl-6-chloro-1-ethyl-2-oxo-1,2-dihydroquinoline-4-carboxylate 3b.
1
Yield (%) = 88; Physical state: Brown crystals; mp =395 K; H NMR (300 MHz,
3
3
3
CDCl ): 1.35 (t, 3H, J = 7.2Hz, CH ), 1.42 (t, 3H, J = 7.2Hz, CH ), 4.34 (q, 2H, J
3
H-H
3
H-H
3
H-H
3
3
=
7.2Hz, CH ), 4.45 (q, 2H, J
= 7.2 Hz, CH ), 7.24 (s, 1H, CH), 7.34 (d, 1H, J = 9.3
H-H
2
H-H
2
3
4
4
Hz, CHarom), 7.53 (dd, 1H, J = 9.3 Hz, J = 2.4 Hz, CHarom), 8.44 (d, 1H, J = 2.7 Hz,
H-H
H-H
H-H
1
3
CHarom). C NMR (75 MHz, CDCl ): 164.80 (C=O), 160.47 (C=O), 137.91-137.35 (Cq-Cq),
3
1
31.06 (CHarom), 128.22 (Cq), 126.83 (CHarom), 125.71 (CHarom), 118.82 (Cq), 115.64
-
1
(
CHarom), 62.16(CH ), 37.84(CH ), 14.10(CH ), 12.57(CH ). IR (ν , cm ): νC -
2 2 3 3 max SP2
-1
-1
-1
-1
H=2959.66cm ; νC -H=2852.37cm ;νC=O=1730.42cm ; νC=C=1660.28-1550.80cm .
SP3
Ethyl-6-bromo-1-ethyl-2-oxo-1,2-dihydroquinoléine-4-carboxylate 3c.
1
Yield (%) = 79; Physical state: Green powder; mp =387 K; H NMR (300 MHz,
3
3
3
CDCl ): 1.35 (t, 3H, J = 7.14Hz, CH ), 1.45 (t, 3H, J = 7.13Hz, CH ), 4.35 (q, 2H, J
3
H-H
3
H-H
3
H-
=
=
3
3
=
7.14 Hz, CH ), 4.45 (q, 2H, J
= 7.14 Hz, CH ), 7.43 (s, 1H, CH), 7.5 (d, 1H, J
2
H
2
H-H
H-H
3
4
4
8
.8 Hz, CHarom), 7.65 ( dd, 1H, J
= 8.8 Hz, JH-H= 2.32 Hz, CHarom), 8.52 (d, 1H, J
H-H
H-H
1
3
2
.3Hz, CHarom). C NMR (75 MHz, CDCl ):164.79 (C=O), 160.47 (C=O), 138.25-137.33
3
(Cq-Cq), 133.84 (CHarom), 129.86 (CHarom), 125.65 (CH), 119.26 (Cq), 115.92 (CHarom),
-1
1
15.67 (Cq), 62.18 (CH ), 37.82 (CH ) 14.11 (CH ), 12.56 (CH ).IR (ν , cm ): νC -
2 2 3 3 max SP2
-1
-1
-1
-1
H=3124.12cm ; νC -H=2975.74cm ;νC=O=1726.58cm ;νC=C=1660.54-1581.59cm .
SP3
2
2
.3. Single crystal X-ray diffraction
.3.1. Crystal structure of 2-oxo-ethyl 1,2-dihydroquinoline-4-carboxylate derivatives 3a
and 3b.
Crystallographic analysis of compounds 3a and 3b obtained by alkylation reaction
thereby alkylating the nitrogen and oxygen at the same time to obtain dialkylated products
and confirms the structures of compounds 3a (Figures 1 and 2) and 3b (Figures 3 and 4). It is
interesting to note that compound 3a and 3b crystallize in the monoclinic system (P21/n). The

ACCEPTED MANUSCRIPT
crystallographic data has been assigned to the deposit number of compound 3a: CCDC
1
833339 and compound 3b: CCDC 1810757.
The title compound, C H NO , 3a exists in an "extended" conformation in the crystal.
14
15
3
Intermolecular C—H···O hydrogen bonds form wavy layers of molecules oriented
approximately parallel to (100). Figure 2).
(
Figure 1: Molecular structure of the compound 3a with the atom-labelling scheme. Displacement
ellipsoids are drawn at the 50 % probability ellipsoids. (CCDC 1833339).
Figure 2: Molecular packing of the compound 3a along the axis with C—H···O interactions shown as
dotted lines.

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Figure 3: Molecular structure of the compound 3b with the atom-labelling scheme. Displacement
ellipsoids are drawn at the 50 % probability ellipsoids. (CCDC 1810757).
Figure 4: Molecular packing of the compound 3b show absence of Pi-Pi interaction and no H binding.
Table 1 Experimental details
Compound 3a
Compound 3b
Crystal data
Chemical formula
Mr
C
14
H
245.27
15NO
3
C H14ClNO
14 3
279.71
Crystal system, space
group
Monoclinic, P2
1
/c
Monoclinic, P2 /n
1
Temperature (K)
100
200
7
.0639 (11), 13.762 (2),
4.3316 (2), 11.6891 (5),
25.8977 (12)
91.291 (2)
1310.93 (10)
4
a, b, c (Å)
1
2.516 (2)
β (°)
V (Å3)
Z
93.272 (2)
1214.8 (3)
4
Radiation type
Mo Kα
Mo Kα

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µ (mm−1)
0.10
0.29
Crystal size (mm)
0.36 × 0.17 × 0.16
Data collection
0.30 × 0.06 × 0.04
Diffractometer
Bruker Smart APEX CCD
Multi-scan
TWINABS (Sheldrick, 2009)
Bruker D8 VENTURE
Multi-scan
SADABS (Sheldrick, V2016/2)
Absorption correction
No. of measured,
independent and
0.81, 0.98
59970, 3841, 3152
observed [I> 2σ(I)
reflections
Rint
]
34779, 5574, 4726
0.078
0.705
(sin θ/λ)max (Å−1)
0.039
Refinement
R[F2> 2σ(F2)], wR(F2), S
No. of reflections
0.048, 0.133, 1.07
5574
0.048, 0.124, 1.08
3841
No. of parameters
224
174
All H-atom parameters refined
H-atom parameters
constrained
0.34, −0.38
H-atom treatment
ꢀρmax, ꢀρmin (e Å−3)
0.45, −0.22
2
.4. Computational details
Density Function Theory (DFT) methods are very important owing to cost of
computations and reliable results for complex molecules in quantum chemistry. In this
method frequentlyBecke-3-Lee-Yang-Parr (B3LYP) are used as functional [33], for
computations 6-31G(d,p) basis set was selected and was used Gaussian 09W software
package[34]. GaussView5 program package [35] was used for visualizations of the computed
structural and vibrational spectroscopic properties. The initial geometry for the molecular
optimization in 3a and 3b compounds were taken from .cif file generated by single crystal X-
Ray diffraction, but 3c compound was computed by taking the structure of 3a and 3b was
based on because we did not get this product as 3c crystals. The computed harmonic
vibrational wavenumbers were scaled with 0.9610 for the B3LYP/6-31G(d,p) level [36], non-
harmonic interactions were not taken in vibrational computations. The theoretical vibrational
assignments of the corresponding compounds (3a, 3b and 3c) using percentage potential
energy distribution (PED) were carried out with VEDA 4 program [37, 38]. To enlighten the
nature of the crystal molecules (intermolecular interactions), Crystal Explorer 3.1 program
was used [39] for Hirshfeld surface analysis. Furthermore, highest occupied molecular
orbital-HOMOs and lowest unoccupied molecular orbital-LUMOs of 3a, 3b and 3c were
1
13
obtained with Chemissian software [40]. The H and C NMR chemical shifts were
calculated with gauge-including atomic orbital (GIAO) approach by using B3LYP/6-31G(d,p)
and compared with the experimental NMR spectra in CDCl -chloroform solvent. Lastly, in
3

ACCEPTED MANUSCRIPT
this work, molecular docking (ligands 3a, 3b and 3cand target receptor) simulations were
performed by using AutoDockVina free software program [41].
2
.4.1. Molecular geometric properties
The compounds-3a, 3b and 3c are not planar. The optimized molecular structures along
with the numbering scheme 1 for 3a, 3b and 3c were shown in Figure 5. The optimized bond
lenghts and bond angles of the title molecules were calculated using the B3LYP functional
with 6-31G(d,p) basis set. Some important and selected Theoretical and experimental
geometric parameters were listed in Table 2. From Table 2, we can see that there are some
mismatchs between experimental and optimized structure. Because optimized structure is in
gas phase, but experimental compounds are in solid phase.
In oxo-dihydroquinoline group, the O1=C9 bond lenghts were calculated as 1.231 Å in
3
a, 1.230 Å in 3b and 1.228 Å in 3c with B3LYP/6-31G(d,p) functional/basis set, these bond
lenghts experimentally were determined as 1.2422 Å, 1.2314 Å and 1.228 Å, respectively.
This bond (C13=O16- according to the their number) was calculated at 1.2232 Å with
B3LYP/6-311++G(d,p) for ethyl-4-hydroxy-2-oxo-1, 2-dihydroquinoline-3-carboxylate [41].
In same group ring N1-C9, N1-C1, N1-C10 and C10-C11 bond lenghts were calculated at
1
.403 Å, 1.396 Å, 1.473 Å and 1.530 Å, respectively in 3a compound ; 1.403 Å, 1.394 Å,
.474 Å and 1.529 Å, respectively in 3b compound ; 1.409 Å, 1.387 Å, 1.476 Å and 1.529 Å,
1
respectively in 3c compound. In literature [42], these bond lenghts are very compatible with
our results. Between oxo-dihydroquinoline group and carboxylate group C7-C12 bond lenghts
were computed at 1.506 Å in 3a compound, 1.506 Å in 3b compound and 1.507 Å in 3c
compound with B3LYP/6-31G(d,p) functional/basis set, and experimentally were determined
as 1.507 Å in 3a compound, 1.5058 Å and 3b compound, but for 3c compound could not be
determined experimental value. In carboxylate group, O2=C12, O3-C12, O3-C13 and C13-
C14 bond lenghts were calculated at 1.216 Å, 1.348 Å, 1.449 Å and 1.516 Å, respectively in
3
a compound ; 1.216 Å, 1.346 Å, 1.450 Å and 1.516 Å, respectively in 3b compound ; 1.215
Å, 1.345 Å, 1.451 Å and 1.515 Å, respectively in 3c compound with B3LYP/6-31G(d,p)
functional/basis set. In ethyl-4-hydroxy-2-oxo-1, 2-dihydroquinoline-3-carboxylate [42],
these bond lenghts were reported as 1.2448 Å (C19=O20-their numbering format), 1.3288 Å
(C19-O21-their numbering format), 1.4500 Å (C22-O21-their numbering format) and 1.5209
Å (C22-C25-their numbering format),respectively. In 3a, 3band3c, between changing atoms
bond lenghts were computed at 1.085 Å (C4-H), 1.558 Å (C4-Cl) and 1.544 Å (C4-Br),
respectively.

ACCEPTED MANUSCRIPT
Secondly, we can look at the some important bond angles in oxo-dihydroquinoline group.
The bond angle C2-C1-C6 and C1-C6-C5 were calculated at 118.9
a ; 118.8 and 119.0 respectively in 3b ; 119.2 and 118.5 respectively in 3c. These values are
lesser than 120 because of the presence of quinoline ring. This situation was determined in
literature. For example, 4-hydroxy-2-oxo-1,2-dihydroquinoline-7-carboxylic acid molecule
43], the bond angle C1-C2-C3 and C2-C3-C4 were reported as 119.6 and 119.4, respectively.
The O1-C9-N1, O1-C9-C8, N1-C9-C8 and N1-C10-C11 bond angles were computed at 121.4
122.9, 115.7 and 112.9, respectively in 3a compound ; 121.5 ,122.9, 115.7 and 112.9
respectively in 3b compound ; 121.1 ,123.1, 115.8 and 112.9, respectively in 3ccompound
with B3LYP/6-31G(d,p) functional/basis set in our study.
In carboxylate group, O2=C12-O3, O2=C12-C7, C12-O3-C13, O3-C13-C14 and C7-
C12-O3 bond angles were computed at 122.8, 125.6, 115.9, 107.4 and 111.7, respectively in
and 111.7, respectively in 3b compound ; 123.1
, respectively in 3c compound with B3LYP/6-31G(d,p)
̊ and 118.68 ̊, respectively in
3
̊
̊
̊
̊
̊
[
̊
̊
̊
,
̊
̊
̊
̊
̊
̊
̊,
̊
̊
̊
̊
̊
̊
̊
̊
̊
3
a compound ; 122.9
̊
̊
̊
̊
̊
̊,
1
25.3, 115.9, 107.4 and 111.6̊
̊
̊
functional/basis set. These bond lenghts and bond angles values are generally compatible with
similar structures in literature [42-45].
Figure 5. The structure of quinoline derivatives 3a, 3b and 3c
Table 2. The selected optimized structure parameters of the title compounds 3a, 3b and 3c
Bond angles (°)
X-Ray
Bond lenghts (Å)
X-Ray
Optimized bond
angles
DFT/B3LYP
6-31 G(d,p)
Optimized
bond lenghts
DFT/B3LYP
6-31 G(d,p)
Compound 3a
C12-O3-C13
O3-C12-C7
O2-C12-C7
O2-C12-O3
O3-C13-C14
116.44(11)
110.81(11)
125.94(13)
123.24(13)
106.46(12)
115.9
111.7
125.6
122.8
107.4
O1-C9
1.2422(17)
1.2114(16)
1.3361(17)
1.4530(17)
1.3742(18)
1.231
1.216
1.348
1.449
1.403
O2-C12
O3-C12
O3-C13
N1-C9

ACCEPTED MANUSCRIPT
O3-C13-H13B
C14-C13-H13B
O3-C13-H13A
O1-C9-N1
O1-C9-C8
N1-C9-C8
N1-C10-C11
C10-C11-H11C
N1-C10-H10A
H(X)-C4-C5
H(X)-C4-C3
C11-C10-H10A
106.8(10)
113.9(10)
108.9(11)
121.64(13)
122.0(13)
116.36(12)
110.93(12)
109.1(18)
109.8(10)
120.9(11)
119.3(11)
111.2(10)
108.6
112.3
108.7
121.4
122.9
115.7
112.9
109.9
108.4
120.0
120.5
111.9
N1-C1
1.3993(16)
1.4795(17)
1.512(2)
1.502(2)
1.507(18)
0.992(19)
1.396
1.473
1.530
1.516
1.506
1.085
N1-C10
C10-C11
C13-C14
C7-C12
C4-X (H)
Compound 3b
C12-O3-C13
O1-C9-N1
O1-C9-C8
115.49(11)
121.87(13)
121.92(13)
116.21(12)
110.2
111.68(13)
109.3
109.5
109.3
112.13
124.83
123.04
107.69
110.17
110.15
119.09
119.58
115.9
121.5
122.9
115.7
108.7
112.9
109.5
109.8
108.4
111.7
125.5
122.9
107.4
112.3
108.5
119.7
119.3
(X)Cl-C4
O1-C9
1.7360(15)
1.2314(17)
1.3246(17)
1.4561(17)
1.1962(18)
1.5058(18)
1.485(2)
1.514(2)
1.4737(17)
1.377
1.758
1.230
1.346
1.450
1.216
1.506
1.516
1.529
1.474
1.403
1.394
O3-C12
O3-C13
O2-C12
C7-C12
C13-C14
C10-C11
N1-C10
N1-C9
N1-C9-C8
O3-C13-H13B
N1-C10-C11
C11-C10-H10A
C10-C11-H11C
N1-C10-H10A
O3-C12-C7
O2-C12-C7
O2-C12-O3
N1-C1
1.397
O3-C13-C14
C14-C13-H13B
O3-C13-H13A
Cl(X)-C4-C5
Cl(X)-C4-C3
Compound 3c
C12-O3-C13
O3-C12-C7
O2-C12-C7
--
--
--
--
--
--
--
--
--
--
--
--
--
--
--
--
--
115.9
111.6
125.3
123.1
107.4
108.5
112.3
108.5
121.1
123.1
115.8
112.9
110.0
108.4
119.7
122.0
112.0
O1-C9
--
--
--
--
--
--
--
--
--
--
--
1.228
1.215
1.345
1.451
1.409
1.476
1.387
1.544
1.515
1.507
1.529
O2-C12
O3-C12
O3-C13
N1-C9
N1-C10
N1-C1
C4-Br (X)
C13-C14
C7-C12
C10-C11
O2-C12-O3
O3-C13-C14
O3-C13-H13B
C14-C13-H13B
O3-C13-H13A
O1-C9-N1
O1-C9-C8
N1-C9-C8
N1-C10-C11
C10-C11-H11C
N1-C10-H10A
H(X)-C4-C5
H(X)-C4-C3
C11-C10-H10A
2
.4.2. Some important Vibrational frequencies

ACCEPTED MANUSCRIPT
From the Figure 5, our compounds have 33 atoms and in nonlinear. For with N atom
system generally, there are 3N-6 modes are vibrational movements or modes. In this study,
we have totally 99 modes and 93 vibrational modes, and for our compounds are in C point
1
group symmetry. In the present study, harmonic vibrational frequencies were computed by
using B3LYP/6-31G(d,p) functional/basis set in gas phase. The vibrational assignments of the
calculated wavenumbers were obtained in terms of potential energy distribution (PED) using
VEDA 4 program [37] and were listed in Table 3-5. In all computational vibration modes
were used corrected scaling factor as 0.9610 [36]. The obtained selected important
frequencies (Unscaled and scaled forms) and their assignments and experimental R spectrum
values consistent with literature [46, 47], were given in Table 3-5.
Table 3. Selected experimental and calculated vibrational modes of 3a
Assignments
Unscaled Scaled Experimental
νCH(98) in the C5-H5
νCH(99) in the C8-H8
νCH(99) in the C2-H2
νCH(97) in the C4-H4
3261
3259
3242
3207
3189
3158
3142
3139
3133
3124
3110
3093
3068
3058
3055
1793
1748
1664
1649
1606
3134
3132
3116
3082
3065
3035
3020
3017
3011
3002
2988
2972
2948
2939
2936
1723
1680
1599
1585
1543
3000-3100.33
νCH(98) in the C3-H3
νCH(92) in the CH2 asym with C10
νCH(84) in the CH2 asym with C13
νCH(96) in the CH3 asym with C11
νCH(100) in the CH3 asym with C14
νCH(92) in the CH3 asym with C11
νCH(85) in the CH2 symm with C13
νCH(95) in the CH2 symm with C10
νCH(99) in the CH2 symm with C13
νCH(99) in the CH3 symm with C14
νCH(90) in the CH3 symm with C11
νC=O(87) in the O2=C12
3
079.63
3025.96
2979.33
2925.69
1717.63
νC=O(68) in the C9=O1
νC=C(43) in the C4=C5 and C7=C8
νC=C(43) in the C4=C5 and C7=C8
νC=C(43) in the C7=C8
1
570.73-1511.83
Table 4. Selected experimental and calculated vibrational modes of 3b
Assignments Unscaled Scaled Experimental
νCH(99) in the C5-H5
νCH(100) in the C8-H8
νCH(91) in the C2-H2
νCH(92) in the C3-H3
νCH(92) in the CH2 asym with C10
νCH(83) in the CH2 asym with C13
νCH(96) in the CH3 asym with C11
νCH(99) in the CH3 asym with C14
νCH(92) in the CH3 asym with C11
νCH(95) in the CH2 symm with C10
νCH(99) in the CH2 symm with C13
3269
3261
3246
3219
3159
3143
3139
3134
3125
3095
3070
3142
3132
3119
3093
3036
3020
3016
3012
3003
2974
2950
3124.12
2
975.74
3056.69
2
959.66

ACCEPTED MANUSCRIPT
νCH(100) in the CH3 symm with C14
νCH(99) in the CH3 symm with C11
νC=O(87) in the O2=C12
νC=O(69) in the C9=O1
νC=C(43) in the C7=C8
νC=C(49) in the C4=C5 and C7=C8
νC=C(43) in the C7=C8
νC-Cl(18) in the C4-Cl
3059
3055
1794
1750
1659
1642
1596
870
2940
2936
1724
1682
1594
1577
1533
836
2
852.37
730.42
1
1
660.28-1550.80
896.91
Table 5. Selected experimental and calculated vibrational modes of 3c
Assignments Unscaled Scaled Experimental
νCH(100) in the C8-H8
νCH(97) in the C2-H2
νCH(99) in the C5-H5
νCH(98) in the C3-H3
3260
3242
3233
3178
3162
3143
3141
3134
3127
3100
3071
3059
3057
1796
1756
1659
1632
1585
1246
3133
3116
3107
3054
3039
3020
3019
3012
3005
2979
2951
2940
2938
1726
1688
1594
3
3
062.92
100.86
νCH(85) in the CH2 asym with C10
νCH(82) in the CH2 asym with C13
νCH(83) in the CH3 asym with C11
νCH(98) in the CH3 asym with C14
νCH(92) in the CH3 asym with C11
νCH(95) in the CH2 symm with C10
νCH(98) in the CH2 symm with C13
νCH(100) in the CH3 symm with C14
νCH(99) in the CH3 symm with C11
νC=O(87) in the O2=C12
2959.66
3124.12
2975.74
1726.58
νC=O(75) in the C9=O1
νC=C(45) in the C7=C8
νC=C(51) in the C7=C8
νC=C(42) in the C4=C5
1568 1660.54-1581.59
1523
1197
νC-Br(25) in the C4-Br
1233.98
2
.4.3. HOMO-LUMO Analysis
The Highest Occupied Molecular Orbital (HOMO) and Lowest Unoccupied Molecular
Orbital (LUMO) parameters are very important to understand quantum chemistry mechanism
and chemical reactions ways. Usually, HOMO is known as donor, while LUMO is known
acceptor [48, 49].While HOMO energy is related to ionization potential, LUMO energy is
related to electron activation. Other important parameter is gap values of title compounds.
The energy differences between HOMO and LUMO are called energy band gap (ꢀE=ELUMO
-
EHOMO) and for the stability of the structures is very crucial. That is the Higher-gap energy is
more stable than smaller-gap situation. By using HOMO and LUMO energy values we can
compute the other important following parameters. Ionization potential=I = -EHOMO ; the
electron affinity =A = -ELUMO ; the chemical hardness= η = (I-A)/2 ; the electronic chemical
2
potential, ꢁ = 1/2(ELUMO+EHOMO) ; the electrophilicity index, ψ = ꢁ /2η, and softness, ζ = 1/η.
These parameters have been calculated and listed in Table 6.
Table 6. HOMO and LUMO Energy Values and other related parameters of 3a, 3b and 3c compounds.

ACCEPTED MANUSCRIPT
Parameters (eV)
ELUMO (eV)
EHOMO (eV)
Energy bandgap |EHOMO-ELUMO
Ionization potential (I = -EHOMO
Electron affinity (A = -ELUMO
3a,
-1.988
-5.958
3.970
5.958
1.988
3b
-2.221
-6.064
3.843
6.064
2.221
3c
-2.377
-5.211
2.834
5.211
2.377
|
)
)
Chemical hardness (h = (I-A)/2)
Chemical softness (z = 1/2h)
Electronegativity (χ = (I+A)/2)
1.985
0.252
3.973
1.922
0.260
4.142
1.417
0.353
3.794
Chemical potential (ꢀ= -(I+A)/2)
Electrophilicity index (w = ꢀ /2h)
-3.973
3.976
-4.142
4.465
-3.794
5.080
2
Sum of Electronic and zero point Energies (a.u)
-822.739916
-1282.3427
-846.872522
The HOMOs and LUMOs figures 6 (HOMO-LUMO cloud distributions) were obtained and
simulated from the optimized molecular geometry using .chk file in gas phase at B3LYP/6-
3
1G(d,p) functional/levels for our compounds in Figure 6. Additionally, the contributions to
HOMOs and LUMOs of three molecular groups, which are group 1, group 2 and group 3 in
our compounds were calculated with Chemissian software [40]. From the output program, we
listed results in Table 7.
Table 7. The contributions to HOMOs and LUMOs of 3a, 3b and 3c
3
a
3b
Group
2
3c
Group
2
MOs
Group
1
Group
2
Group
3
Group
1
Group
3
Group
1
Group
3
HOMO %4
LUMO %3
%96
%77
%0
%20
%3
%3
%97
%78
%0
%19
%0
%3
%100
%79
%0
%18
Group 1 : C10-C11, H10A,H10B, H11A, H11B, H11C
Group 2 : C1-C9, N1, O1, H2-H3-X-H5,H8
Group 3 : O2-O3, C12-C14, H13A,H13B, H14A,H14B,H14C

ACCEPTED MANUSCRIPT
Figure 6. HOMO and LUMO plots of 3a, 3b and 3c compounds in B3LYP/6-31G(d,p).
From Figure 6 and Table 7, both the HOMOs and the LUMOs were mainly focused on
Group 2. This group constitutes more than half the HOMO-LUMO distributions. These
contributions point out that the abundance of the electron localizations on Group 2 of the title
compounds can be indicated the electronic transitions and charge transfers in this group.
2
.4.4. NMR Analysis
The content and purity of a compound can be determined using NMR spectroscopy. So as
a spectroscopy technique NMR is very important in both organic an inorganic chemistry and
1
13
in theoretical computations [50, 51]. The theoretical H and C NMR chemical shifts were
computed in chloroform (CDCl3) solvent of the title compounds at the B3LYP/6-31G(d,p)
level using IEFPCM model and GIAO method. The experimental and computed proton and
carbon-13 NMR chemical shifts were given in Table 8 and Table 9. Generally, the observed
1
13
and calculated H and C NMR chemical shifts are in a very good agreement with the
literature [43, 44]. There are fourteen carbon-13 and fifteen proton NMR chemical shifts for
the title molecules that we can locate precisely as seen Table 8 and 9.
1
3
Table 8. The experimental and computed C NMR isotropic chemical shifts (with respect to TMS, all
values in ppm) of 3a, 3b and 3c
Atoms
3a
3b
3c
δexp.
δcal.
δexp.
δcal.
δexp.
δcal.
C1
C2
C3
C4
C5
C6
C7
C8
139.36 129.89 137.91 128.68 138.32 134.23
114.35 102.59 115.64 103.85 115.92 102.74
131.07 119.19 131.06 119.26 133.32 118.15
122.45 110.21 128.22 125.22 119.26 126.11
127.37 117.19 126.83 116.83 129.86 121.42
117.80 108.63 118.82 109.86 119.26 108.14
138.90 127.92 137.35 126.27 137.33 128.27
124.20 114.91 125.71 116.58 129.86 116.06
160.95 147.32 160.47 147.08 160.47 147.97
C9
C10
C11
C12
C13
C14
37.64
12.63
32.78 37.84
6.47 12.57
33.04 37.82
6.29 12.56
33.39
6.44
165.46 154.35 164.80 153.87 164.79 153.73
61.99
14.38
56.21 62.16
7.18 14.10
56.36 62.18
7.12 14.11
56.71
7.13
1
Table 9. The experimental and computed H NMR isotropic chemical shifts (with respect to TMS, all
values in ppm) of 3a, 3b and 3c
Atoms
3a
3b
3c
δexp.
δcal.
δexp.
δcal.
7.66
7.77
δexp.
7.50
7.65
δcal.
7.75
8.29
H2
H3
7.45 7.74 7.34
7.62 7.98 7.53
H4
7.30 7.64 With Cl With Cl With Br With Br
H5
H8
H10A
8.35 9.11 8.44
7.20 7.46 7.24
4.40 3.88 4.34
9.21
7.57
3.82
8.52
7.43
4.35
9.67
7.61
3.92

ACCEPTED MANUSCRIPT
H10B
H11A
H11B
H11C
H13A
H13B
H14A
H14B
H14C
4.40 5.17 4.34
1.39 1.62 1.35
1.39 1.66 1.35
1.39 1.34 1.35
4.46 4.67 4.45
4.46 4.60 4.45
1.42 1.80 1.42
1.42 1.49 1.42
1.42 1.74 1.42
5.14
1.60
1.35
1.60
4.67
4.61
1.76
1.80
1.48
4.35
1.35
1.35
1.35
4.45
4.45
1.45
1.45
1.45
5.17
1.65
1.42
1.65
4.77
4.69
1.79
1.84
1.54
2
.5. Hirshfeld surface calculations
Crystal Explorer 3.1 program [39] was used for Hirshfeld surface analysis calculations
and evaluations. With this analysis, we understand the nature of the crystal molecules and we
determine important interactions between the molecules. For computations, *.cif file of the
crystal is used initially. The Hirshfeld surface is obtained for scanning the van der Waals
distances and to define the interaction sites [52]. In this analysis, two parameters are describe
when obtaining a molecule Hirshfeld surface. d is distance from the surface of the Hirshfeld
e
to the nearest core outside the surface; and d : distance corresponding to the nearest core in
i
the surface. dnorm value is dependent on d , d and Van der Walls radius, that is these
e
i
expressions could be expressed as follows :
ꢅꢆꢇꢃ ꢆꢈ ꢉꢊ
ꢃ ꢆꢈ ꢉꢊ
ꢅꢋꢇꢃ ꢋꢈ ꢉꢊ
ꢀꢁꢂꢃꢄ
=
+
(1)
ꢈꢉꢊ
ꢃ
ꢋ
Where the ꢌꢍꢎꢏꢐand ꢌꢑꢎꢏꢐare the Van der Waals radii of the atoms. Additionally, we
canstate that, 3D Hirshfeld surface can be converted into 2D fingerprint plots using by d and
i
de distances values [53]. In the surface there are three color (red, green and blue). Red color
defines contact points shorter than sum of van der Waals radii with negative dnorm, blue color
is contact sites longer than sum of van der Waals radii with positive dnorm; and white color
corresponds to the intermolecular distances close to van der Waals radii with dnorm equal to
zero [53].
In this study 3D Hirshfeld surfaces (dnorm) and 2D fingerprint histograms were
obtained and given in Figure 7 and 8 for 3a and 3b compounds, respectively. In the surface of
Hirshfeld (dnorm), dark red dots show stronger hydrogen bonds than open red dots as seen Fig.
3
, showing surfaces that have been mapped over a dnorm range of -0.2797 (red color) to 1.1987
(blue color) Å for 3a, and dnormrange of -0.2869 (red color) to 1.2009 (blue color) Å for 3b.
As seen Figure 5, the most concentrated red regions are located on the oxo-dihydroquinoline
group. It is seen to the Figure 5 and 6 the contact distances between hydrogen and other atoms
(C, O, Cl and N).

ACCEPTED MANUSCRIPT
Figure 7. The 3D Hirshfeld surface of 3a and 3b compounds.
Figure 8. The 2D fingerprint histogram of 3aand 3b compounds.
2
.6. Molecular Docking Studies
Molecular docking is very crucial way to understanding of the binding interactions between
small molecule (a ligand) and protein receptor. This method for drug design is a highly
reliable program, cost-effective and time-saving [54, 55]. 3a, 3b and 3c compounds are
quinoline heterocycles and these type of molecules have profound medicinal and
pharmacological importance, so we have undertaken in silico based molecular docking study.

ACCEPTED MANUSCRIPT
But in this study, we could not performe docking process for 3c compound. Because bromine
atom is the third-lightest halogen, and blind docking requires. Therefore, we continued for
compounds 3a and 3b ligands. Initially, we optimized the structure with B3LYP/6-31G(d,p)
functional/basis set and the optimized structure for ligand molecules were composed in PDB
format (Protein Data Bank). Later, to determine the target, optimized ligand structure in PDB
was subjected to Swiss Target Prediction [56], also related literature was used [57]. We
obtained target protein 1M17-EGFR kinase. This target protein structure was taken from the
Protein Data Bank [58]. The water molecules, co-factors and ligands in the target structure
were removed and polar hydrogen atoms were added, later rotatable bonds and torsion tree
root were determined. Both ligand and target structures were prepared with Discover Studio
Visualizer 4.0 (DSV 4.0) software [59]. Molecular docking computations for 3a and 3b were
performed with AutoDockVina program [41]. For 3a-1M17 and 3b-1M17 complex molecular
3
3
docking positions were determined as 108x80x66 Å and 92x60x56 Å and obtained surfaces
around ligand and 2D forms of molecular positions like Figure 9 (a,b) Figure 10 (a,b),
respectively. According to the rotable bonds of ligand, nine modes were superposed and their
affinity energy values, root mean square (RMS) values were calculated and given in Table 10.
Table 10. AutoDockVina results of the binding affinity and RMSD values of different poses in 1M17
inhibitor of 3a and 3b compounds.
3a-1M17
3b-1M17
Mode
affinity
kcal/mol)
rmsdl.b. rmsdu.b.
affinity
(kcal/mol)
-7.8
-7.0
-6.9
-6.6
-5.8
-5.5
-5.4
rmsdl.b. rmsdu.b.
(
1
2
3
4
5
6
7
8
9
-7.4
-7.1
-6.5
-6.5
-5.7
-5.7
-5.6
-5.6
-5.4
0.000
1.650
2.038
2.425
16.344
4.005
3.835
4.896
16.211
0.000
2.183
4.957
5.166
18.602
6.355
4.834
7.781
18.437
0.000
3.184
2.072
1.911
3.098
30.862
7.657
17.901
19.689
0.000
5.776
4.843
4.841
5.350
32.565
10.014
20.040
21.055
-5.3
-5.2
From the literature, a good docking results will have RMSD values nearer to 2Å [60]. From
the Table 10, we can say that the best docking pose was between ligands (3a and 3b) and
1
M17, with the binding energy of -7.8 kcal/mol. The 3a-1M17 and 3b-1M17 docking results
and their interaction amino acids could be easily seen from the Figure 9 and 10. Finally, we
-6
-6
computed the inhibition constant (K ) as 1.91672x10 M for 3a compoundand3.76496 x10
i
M for 3b compound from K =exp(ꢀG/RT) equation, where ꢀG, R and T are the docking
i
-3
energy (binding free energy), gas constant (1.9872036×10 kcal/mol) and room temperature
298.15 K), respectively.
(

ACCEPTED MANUSCRIPT
Figure 9. The molecular docking results of the 3a compound with 1M17 protein, surfaces around ligand (a)
and 2D forms (b).
Figure 10. The molecular docking results of the 3b compound with 1M17 protein, surfaces around ligand
(a) and 2D forms (b).
3
3
. Study of Antioxidant Properties
. 1. Free radical scavenging activity
The free radical scavenging activity of our compounds was evaluated according to the method
proposed by Jeong et al. [61] with some modifications. 1 mL aliquot of the three compounds
with various concentration dissolved in ethanol, was mixed with 0.9 mL of compound 2,2-
diphenyl-1-picrylhydrazyl (DPPH) (0.041 mM). The optical density of the samples was
measured at 517 nm. Butylated hydroxytoluene (BHT) solution was used as a standard. The
radical scavenging activity was expressed as percentage inhibition by the following equation:

ACCEPTED MANUSCRIPT
-1
IC50 value (mg.mL ) is the concentration of the sample which corresponds to 50%
inhibition of DPPH in solution.
3
. 2. Reducing power
The reducing power was determined according to the method of Bougandoura et al. [62]. 1
mL of our compounds (1-10 mg/mL) were mixed with phosphate buffer (2.25 mL, 0.2 M, pH
6
.6) and potassium ferricyanide [K Fe(CN) ] (2.25 mL, 1%). After incubation of the mixture
3 6
at 50°C for 20 min, 2.25 mL of trichloroacetic acid (10%) was added and the tubes undergo a
centrifugation at 3000 rpm for 10 min. The upper layer of the solutions (2.25 mL) were mixed
with distilled water (2.25 mL) and FeCl (0.5 mL, 0.1%) and the absorbance of the reaction
3
mixtures at 700 nm indicated increased reducing power. BHT was used as a positive control.
3
. 3. Inhibition of β-carotene bleaching
Lipid peroxidation inhibition (LPI) activity was evaluated according to the β-carotene
bleaching method of Boudjou et al. [63]. 1 mL of β-carotene (0.2 mg/mL) dissolved in
chloroform was added to a round-bottom flask containing linoleic acid (40 µl) and Tween-80
(400 µl). After removing the solvent, 100 mL of demineralized water were added. 5 mL of
this mixture were added to 0.5 mL of every one of our compounds (1mg/mL). MeOH 80%
was used as control or BHT as standard. The absorbance was measured immediately (t=0) and
periodically after 15 min, against a blank at 470 nm, until the end of the experiment (t = 120
min). LPI activity was expressed as AOA (%) and was calculated as follows: The method of
calculation was based on first order kinetics as described by Al-Saikhan et al. [64] using the
following equations Bleaching rate ofβ-carotene :
Where A is the initial absorbance (470 nm) of the emulsion at time 0, while a its the
0
t
absorbance (470 nm) at 15, 30… and 120 min; and t is the time in min. The rates for different
times were averaged to give one value for the sample. The antioxidant activity was expressed
based on the oxidation rate ratio, ORR, which was calculated by the method of Marinova et
al. [65] using the equation:
BR is the rate of β-carotene bleaching in the presence of the sample, and BR is the rate of
s
o
bleaching in the absence of the sample (blank). The antioxidant activity (AOA%) was
calculated as % inhibition relative to the control using the following relationship [66]:

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Table 11: Antioxidant activity of compounds
β-carotene
bleaching
DPPH
FRAP
Compounds
IC50 -
IC50
IC50
mg/ml
IC50
mgBHTeqv.mg
AOA%
1
-1
-1
mg.mL
mgBHTeqv.mg
0.029±0.001
0.097±0.002
0.071±0.003
-
3a
3b
3c
0.620±0.32
0.185±0.01
0.255±0.01
0.018
-
77.00
76.69
59.21
99.94
0.82±0.01
1.7±0.15
0.04
0.049±0.001
0,024±0.002
-
BHT
The antioxidant activity was evaluated by three methods: radical scavenging activity, ferric
reducing and β-carotene bleaching. In the two methods radical scavenging activity and ferric
reducing, the IC values denote the concentration of sample, which is required to scavenge
5
0
3
+
2+
5
0% of DPPH free radicals or the ability of the substance to reduce Fe to Fe . DPPH and
FRAP test results were expressed as BHT (butylated hydroxyl toluene) equivalent per one
-1
gram of our compounds (mg BHT eqv.mg ).
There was a significant difference in the accuracy of the best oxidant among the three
methods and the radical scavenging activity was much higher than the ferric reducing
antioxidant activity. The radical scavenging activity had its highest value with the product
(3b) by IC equal to 0.185±0.01 mg/ml and 0.097±0.002 mg BHT/mg followed by product
50
(3c) and product (3a) showing an IC50 of 0.071±0.003 and 0.029±0.001mg BHT/mg,
9
8
7
6
5
4
3
2
1
0%
0%
0%
0%
0%
0%
0%
0%
0%
0
%
0
0.2
0.4
0.6
0.8
1
1.2
Product 3a
Product 3b
Product 3c
respectively (

ACCEPTED MANUSCRIPT
Figure ). Those results was confirmed by the ferric reducing antioxidant activity where the
highest value was 0.049±0.001 mg BHT/mg referring to product (3b) followed by product
(
3c) with IC equal to 0.024±0.002 mg BHT/mg but we cannot reach the IC of product (3a)
5
0
5
0
with the used concentration (Figure 12).
In the case of β-carotene bleaching, β-carotene is attacked by peroxyl radicals that result from
thermal-induced oxidation of linoleic acid. Form Figure 13, we noticed that product (3a) had
highest capacity to reduce bleaching showing the highest AOA 77.00% followed by product
(3b) 76.69 % and product (3c) 59.21%. Those results showed that product (3a) and (3b) had
the same antioxidant activity (77.00% and 76.69%).
9
8
7
6
5
4
3
2
1
0%
0%
0%
0%
0%
0%
0%
0%
0%
0
%
0
0.2
0.4
0.6
0.8
1
1.2
Product 3a
Product 3b
Product 3c
Figure 11. Radical scavenging activity for the three studied products
0
0
0
0
0
0
0
0
.7
.6
.5
.4
.3
.2
.1
.0
0
0.5
1
1.5
Product 3b
2
2.5
Product 3a
Product 3c
Figure 12. Ferric reducing power for the three studied products

ACCEPTED MANUSCRIPT
BHT
Product 3a
MeOH
Product 3b
3
2
1
0
.5
3
Product 3c
.5
2
.5
1
.5
0
0
20
40
60
80
100
120
140
Figure 13. The β-carotene kinetic blanching
4
. Conclusions
In this work we report the synthesis of a new 2-oxo-1,2-dihydroquinoline-4-carboxylic derivatives:
a-3c. Along with its characterization, using X-Ray crystallographic and spectroscopic techniques
are described. Theoretical approach used in this work allow a relatively well reproduction of X-Ray
3
1
13
geometrical coordinates parameters and spectral data and H and C NMR chemical shifts Hirshfeld
surfaces are employed to confirm the existence of intermolecular interactions in the title compounds
3a-3c. The experimental spectroscopic data are well reproduced using DFT and Molecular-docking
study reveals that the title compound 3a and 3b docked. The compounds 3a-3c shows the antioxidant
activity assays to act as potent antioxidants.
Acknowledgements: Author would like to thank Prof. Dr. Fatih Ucun for his helpful contribution
and Gaussian Calculations.
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