Spectroscopic, thermal, catalytic and biological studies of Cu(II) azo dye complexes

Article abstract of DOI:10.1016/j.molstruc.2017.03.082

New complexes of copper(II) with azo compounds of 5-amino-2-(aryl diazenyl)phenol (HL_n) are prepared and investigated by elemental analyses, molar conductance, IR,¹H NMR, UV–Visible, mass, ESR spectra, magnetic susceptibility measure

Full text of DOI:10.1016/j.molstruc.2017.03.082

Accepted Manuscript
Spectroscopic, thermal, catalytic and biological studies of Cu(II) azo dye complexes
A.Z. El-Sonbati, M.A. Diab, A.A. El-Bindary, A.F. Shoair, M.A. Hussein, R.A. El-Boz
PII:
S0022-2860(17)30370-8
10.1016/j.molstruc.2017.03.082
MOLSTR 23578
DOI:
Reference:
To appear in: Journal of Molecular Structure
Received Date: 17 December 2016
Revised Date: 20 March 2017
Accepted Date: 21 March 2017
Please cite this article as: A.Z. El-Sonbati, M.A. Diab, A.A. El-Bindary, A.F. Shoair, M.A. Hussein, R.A.
El-Boz, Spectroscopic, thermal, catalytic and biological studies of Cu(II) azo dye complexes, Journal of
Molecular Structure (2017), doi: 10.1016/j.molstruc.2017.03.082.
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Spectroscopic, thermal, catalytic and biological studies of Cu(II) azo dye
complexes
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1
,+
1
1
1
2
A.Z. El-Sonbati , M.A Diab , A.A. El-Bindary , A.F. Shoair , M.A. Hussein and
1
R.A. El-Boz
1
2
Department of Chemistry, Faculty of Science, Damietta University, Damietta, Egypt
Department of Chemistry, Faculty of Science, Port Said University, Port Said, Egypt
Abstract
New complexes of copper(II) with azo compounds of 5-amino-2-(aryl
diazenyl)phenol (HL ) are prepared and investigated by elemental analyses, molar
n
1
conductance, IR, H-NMR, UV-Visible, mass, ESR spectra, magnetic susceptibility
measurements and thermal analyses. The complexes have an square planar structure and
general formula [Cu(L )(OAc)].H O. Study the catalytic activities of Cu(II) complexes
n
2
toward oxidation of benzyl alcohol derivatives to carbonyl compounds were tested using
H O as the oxidant. The intrinsic binding constants (K ) of the ligands (HL ) and Cu(II)
2
2
b
n
complexes (1-4) with CT-DNA are determined. The formed compounds have been tested for
biological activity of antioxidants, antibacterial against Gram-positive (Staphylococcus
aureus) and Gram-negative (Escherichia coli) bacteria and yeast Candida albicans.
Antibiotic (Ampicillin) and antifungal against (Colitrimazole) and cytotoxic compounds
HL , HL , HL and complex (1) showed moderate to good activity against S. aureus, E. coli
1
2
3
and Candida albicans, and also to be moderate on antioxidants and toxic substances.
Molecular docking is used to predict the binding between the ligands with the receptor of
breast cancer (2a91).
Keywords: Azo compound, Cu(II) complexes; DNA binding; Biological activity; Thermal
analysis.
--------------------------------------------
+
Corresponding author: E-mail address: elsonbatisch@yahoo.com (A.Z. El-Sonbati).
1. Introduction
It has already been a number of organic dyes unified innovations of our day-to-
day life. Azo dyes are hardly organic compounds biodegradable because of its high
stability to light and resistant to microbial attack. The greater part of the azo
compounds are a class of synthetic aggravates that are continuously getting
consideration in experimental exploration [1,2]. Azo dyes can supply a complete
rainbow of colors; therefore they have tremendous importance as dyes and also as
1

pigments for a long time [3]. Azo dyes are generally synthesized by the reaction of
primary aromatic amines by diazotization and coupling with phenol or secondary
ACCEPTED MANUSCRIPT
aromatic amines. In fact, about half of the dyes in the industry are also compounds,
which are mostly prepared from diazonium salts [4-6]. Azo compounds as organic
dyes have attracted considerable attention due to their applications [7,8].
Azo dye compounds coordinate with metal ions depend on the nature of the metal,
their valence, the number of donor atoms within the ligand, the type of chelating rings
formation and pH of the reaction medium. It is known that azo dye compounds and their
metals complexes are known to be involved in a number of biological reactions, such as the
inhibition of DNA, RNA, synthesized protein, nitrogen fixation and carcinogenesis [9-11].
Herein, we describe the synthesis and characterization of 5-amino-2-(aryldiazenyl)
phenol (HL ) and their Cu(II) complexes by different spectroscopic techniques. Calf thymus
n
DNA-binding activity of the ligands and copper(II) complexes were studied by absorption
spectra. Catalytic oxidation of benzyl alcohol derivatives to the corresponding carbonyl
compounds in presence of the formed complexes using H O as oxidant was studied. The
2
2
optimal bond lengths and bond angles has been investigated and calculated.
2
. Experimental
.1. Materials
2
3
-Aminophenol obtained from Aldrich Chemical Company was used without any
further purification. Aniline (99.5%), 4-derivatives anilines (alkyl: CH (96.0%), Cl (98.0%)
3
and NO (97.0%); and copper acetate (CuOAc.H O) from Sigma. All other used chemicals
2
2
and solvents were of analytical reagent grade.
2.2. Synthesis of the ligands and their complexes
2.2.1. Synthesis of the azo dye ligands (HLn)
The ligands of 5-amino-2-(aryldiazenyl) phenol (HL ) were prepared by dissolving
n
aniline or its p-substituted derivatives (10 mmol) in conc. H SO diazotized compound
2
4
below -5 ° C in an ice - salt bath with sodium nitrate solution (0.8 g 0.10 mmol and 30 mL
distilled H O). Diazonium salt was coupled with an alkaline solution of 3- aminophenol (1.0
2
g 0.10 mmol) in 20 ml of ethanol. The precipitate was filtered and dried after through rinsing
with water and ethanol mixture. The crude product was recrystallized from ethanol and the
microcrystals were obtained in a yield of 94-98 %. The resulting formed ligands (Fig. 1) are:
HL = 5-amino-2-(p-methyl phenyl diazenyl) phenol.
1
HL = 5-amino-2-(phenyldiazenyl) phenol.
2
HL = 5-amino-2-(p-chlorophenyl diazenyl) phenol.
3
HL = 5-amino-2-(p-nitro phenyl diazenyl) phenol.
4
2

2.2.2. Synthesis of Cu(II) complexes (1-4)
Copper(II) complexes were prepared (Fig. 2) according to the general procedures
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described by El-Sonbati et al. [8,12]. A stoichiometric amount of the desired ligand (0.02
mol) in ethanol (20 mL) was added dropwise to a hot ethanol solution (20 mL) of
CuOAc.H O (0.01 mol) and the reaction mixture was refluxed for 6 hours. The solution was
2
concentrated and the microcrystalline solid separated, which was isolated by filtration,
washed with hot ethanol, ether and dried in a vacuum dryer over anhydrous CaCl2.
2.2.3. DNA binding experiments
Binding characteristics of ligands and CT-DNA complexes have been studied using
electronic absorption spectroscopy. The stock solution of CT-DNA was prepared in 5 mM
Tris-HCl/50 mM NaCl buffer (pH=7.2), which a ratio of UV absorbance’s at 260 and 280
nm (A /A ) of ca. 1.8-1.9, showing that the DNA was adequately free of protein [13, 14].
2
60 280
-1
-1
Furthermore, the fixation was dictated by UV absorbance at 260 nm (Ɛ= 6600 M cm )
[
15]. Electronic absorption spectra (200-700 nm) were carried out using 1 cm quartz
o
-3
-1
cuvettes at 25 C by fixing the concentration of ligand or complex (1.00×10 mol L ) with
,
-4
-1
a gradual increase in the concentration of CT-DNA (1.30×10 mol L ). The intrinsic
binding constant K of the compound with CT-DNA was determined using the following
b
equation [8,16]:
[
DNA] / (є –є ) = [DNA] / (є –є ) + 1 / K (є –є )
a f b f b a f
Where [DNA] is the concentration of CT-DNA in base pairs, є is the extinction coefficient
a
observed for the A /[compound] at the given DNA concentration, є is the extinction
obs
f
coefficient of the free compound in solution and є is the extinction coefficient of the
b
compound when fully bond to DNA. In plots of [DNA]/(є –є ) versus [DNA], K is given
a
f
b
by the ratio of the slope to the intercept.
2.2.4. Catalytic oxidation of alcohols by Cu(II) complex
The complex [Cu(L )(OAc)] H O (0.0064 g, 0.01 mmol) was dissolved in DMF (2 mL)
1
2
and H O (10 mL), then benzyl alcohol (2 mmol) was added with stirring for 30 min.
2
Hydrogen peroxide (2.3 mL, 30%, 10 mmol) was then added dropwise and reaction mixture
was irradiated ultrasonically for further 30 min then reduced in vacuum. The carbonyl
compound extracted by diethyl ether (3×10 mL), filtered through a bead of silica gel and
dried over anhydrous MgSO . The produced benzaldehyde were quantified as 2,4-
4
dinitrophenylhydrazone derivative.
2.2.5. Biological Studies
2.2.5.1 Antioxidant activity screening assay ABTS method
3

-
For each of the investigated ligands (HL ) and their complexes, 2 mL of 2,2 - azino-bis
n
(3-ethylbenzthiazoline-6-sulphonic acid (ABTS) solution (60 µM) was added to 3 mL
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MnO solution (25 mg/mL) in 5 mL aqueous phosphate buffer solution (pH 7, 0.1 M). The
2
mixture was shaken, centrifuged, filtered, and the average absorbance of the resulting green
blue solution at 734 nm was adjusted to about ca. 0.5. Then, 50 µl of 2 mM solution of test
compound in spectral grade MeOH / phosphate buffer (1: 1) was added. The absorbance
measurement at 734 nm decrease in color intensity was expressed as percentage of inhibition
[
17].
2.2.5.2. Antibacterial activity
Chemical compounds were individually tested against a gram positive (Staphylococcus
aureus), gram negative (Escherichia coli) bacteria and yeast (Candida albicans). 1 mg of
each compound was dissolved in 1 mL DMSO and the solution was placed in petri dish
containing a sterilized Whatman filter paper (5 cm) and media nutrient agar (beef extract 3
g meat + peptone 5 g and 20 g agar) seeded with the tested microorganisms. The petri
dishes were incubated at 35˚C and the inhibition zones were recorded after 24 h. Each
treatment was replicated three times. The antibacterial activity of standard antibiotic
(ampicillin) and antifungal (colitrimazole) was also recorded using the same procedure as
above at the same concentration and solvents. The % activity index for the complex was
calculated by the formula as follows:
%
Activity index = Zone of inhibition by test compound (diameter)
2.2.5.3. Antitumor activity
The cell lines hepatocellular carcinoma (HePG-2) and mammary gland breast cancer
(
MCF-7) were used to determine the inhibitory effects of compounds on cell growth using 3-
4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Solutions of
[
MTT, dissolved in medium or balanced salt solutions without phenol red, are yellowish in
color. Mitochondrial dehydrogenases of viable cells cleave the tetrazolium ring, yielding
purple formazan crystals which are insoluble in aqueous solutions. Cell lines were cultured
in with 10% fetal bovine serum. Antibiotics added were 100 units/ml penicillin and 100
o
µg/mL streptomycin at 37 C in a 5% CO incubator. The cell lines were seeded in a 96-well
2
4
plate at a density of 1.0x10 cells/well for 48 h under 5% CO . After incubation, the cells
2
were treated with different concentration of compounds and incubated again for 24 h. After
2
4 h of drug treatment, 20 µL of MTT solution were added from concentrations range 1.56-
00 µg/mL and incubated for 4 h. Dimethyl sulfoxide (DMSO) in volume of 100 µl is added
1
4

into each well to dissolve the purple formazan formed. The colorimetric assay is measured
and recorded at absorbance of 570 nm using a plate reader (EXL 800). The relative cell
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viability percentage was calculated as (A570 of treated samples/A570 of untreated sample)
×
100 [18].
2.2.6. Molecular docking
Docking calculations were carried out using Docking Server. The MMFF94 force
field was used for energy minimization of ligands molecule using Docking Server. Gasteiger
partial charges were added to the ligand atoms. Non-polar hydrogen atoms were merged,
and rotatable bonds were defined.
Docking calculations were carried out on 2a91 – SIGNALING PROTEIN,
TRANSFERASE, MEMBRANE PR protein model. Essential hydrogen atoms, Kollman
united atom type charges, and solvation parameters were added with the aid of AutoDock
tools. Affinity (grid) maps of 20×20×20 Å grid points and 0.375 Å spacing were generated
using the Autogrid program. AutoDock
Parameter set- and distance-dependent dielectric functions were used in the calculation of
the
van
der
Waals
and
the
electrostatic
terms,
respectively.
Docking simulations were performed using the Lamarckian genetic algorithm (LGA)
and the Solis & Wets local search method. Initial position, orientation, and torsions of the
ligand molecules were set randomly. Each docking experiment was derived from 10
different runs that were set to terminate after a maximum of 250000 energy evaluations. The
population size was set to 150. During the search, a translational step of 0.2 Å, and
quaternion and torsion steps of 5 were applied [19-22].
2.2.7. Analytical techniques
2.2.7.1. Infrared spectroscopy (IR)
-1
IR spectra (KBr discs, 4000-400 cm ) by recorded in Jasco FTIR-4100
spectrophotometer.
1
1
2
.2.7.2. H- Nuclear magnetic resonance spectroscopy ( H- NMR)
1
The H- NMR spectra were recorded by Bruker WP 300 MHz using DMSO-d as a
6
solvent containing TMS as the internal standard.
2.2.7.3. UV–Visible spectroscopy
UV–Visible spectra were recorded by Perkin-Elmer AA800 spectrophotometer Model
AAS.
5

2.2.7.4. Magnetic susceptibility measurements
Magnetic susceptibility measurements has been identified at room temperature on the
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balance of the magnetic susceptibility Johnson Matthey using mercury Hg[Co(SCN) ] as
4
calibration.
2.2.7.5. Thermogravimetric analysis (TGA)
TGA of the ligands and copper complexes were carried out using Shimadzu
o
thermogravimetric analyzer under nitrogen atmosphere with a heating rate of 15 C / min at
a temperature ranging from room temperature up to 800 °C.
2.2.7.6. Microanalysis
Elemental microanalyses of the compounds for C, H, and N were determined on
Automatic Analyzer CHNS Vario ELIII, Germany. The analyses were repeated twice to
check the accuracy of the analyzed data. The metal content in the complexes was estimated
by standard methods [12].
2.2.7.7. X-ray diffraction analysis
X-ray diffraction analyses of the powder form of Cu complexes were performed [23] at
room temperature by a Philips X-ray diffractometer equipped with utilized monochromatic
Cu K radiation (
λ
= 1.540598 Å). The X-ray tube voltage and current were 40 kV and 30
α
mA, respectively.
2.2.7.8. Molecular structure investigations
The molecular structures of the investigated compounds were optimized by HF method
with 3-21G basis set. The molecules were built with the Perkin Elmer ChemBio Draw and
optimized using Perkin Elmer ChemBio 3D software [24].
2.2.7.9. ESR measurements
ESR measurements of powdered samples were recorded at room temperature using an
X-band spectrometer utilizing a 100 kHz magnetic field modulation with diphenyl
picrylhydrazyle (DPPH) as a reference material. The conductance measurement was
achieved using Sargent Welch scientific Co., Skokie, IL, USA.
3. Results and discussion
1
The H NMR spectrum of HL was recorded in DMSO-d solution using TMS as
2
6
internal standard (Fig. S1). The compound under investigation show a signal at δ 10.00 ppm,
which can be assigned to the phenolic –OH group proton with integration equivalent to one
proton. The compound shows a group of multiple-signals corresponding to the aromatic
protons
δ5.95-6.35 and
δ7.35-7.80 ppm. Also, the spectrum show a single at
δ 6.62 ppm (s;
2
H, NH ) due to the two protons of amino group in
p
-position to the N=N group. The ligand
2
show a signal at
δ12.45 ppm, which may be attributed to the presence of intramolecular
6

hydrogen bond between the phenolic –OH group of the moiety in HL and the azodye group.
2
on addition of D O the intensities of both OH and NH protons significantly decrease (Fig.
2
2
ACCEPTED MANUSCRIPT
S1(b)). The peak at (10.30/12.45) ppm, which is due to the exchangeable hydrogen-bonded
hydroxyl (OH) proton, disappears upon exchange with D O and can be associated with the–
2
OH proton involved in intramolecular hydrogen bonding with the azodye nitrogen atom
[
25].
Analytical data indicates that the diazo coupling reaction between 3- aminophenol and
aniline or its p-substituted derivatives occurs in 1 : 1 molar ratio and the product form well
defined complexes with Cu(OAc) .2H O. Formation of the complexes can be represented by
2
2
the following equation.
Cu(OAc) .2H O + HL
→
[Cu(L )(OAc)]H O + CH COOH
1-4 2 3
2
2
1-4
Formulation of the complexes (1-4) has been based on their elemental analytical data,
molar conductance, magnetic susceptibility data and different spectroscopic techniques. The
results are consistent with 1 : 1 metal:ligand stoichiometry (Table 1). The azo dye ligands
and their complexes are soluble in all common organic solvents, non-hygroscopic. The
-3
conductance measurements, recorded for 10 M solutions of the complexes in DMSO show
that copper(II) complexes are non-electrolytes. Hence the one acetate in copper(II)
complexes are in the coordination sphere and confirmed by analytical data.
The values of the yield (%) and the melting point are related to the nature of the para
substituent as they increased according to the following order: p-(NO > Cl > H > CH ).
2 3
This can be attributed to the fact that the charge effectively increased due to the electron
withdrawing substituent (HL ), while it decreased by electrons donating character of HL ,
4
1
HL and HL ligands. This is in accordance with that expected from Hammett’s constant
3
2
R
R
(σ
) as shown in (Fig. S2(a,b)) correlate the yield (%) and melting point values with σ , it
R
is clear that all these values increase with increasing
σ
.
3.1. IR spectra
The IR spectra of the complexes are compared with those of the free ligands (HL ) in
n
order to determine the coordination sites that may be involved in chelation. These peaks,
such as those of OH, N=N, and NH , are expected to be involved in chelation. The position
2
and/or the intensities of the peaks are expected to be changed upon chelation. On examining
these spectra by comparison with the free ligands, one can conclude the following, all
-1
complexes under investigation exhibit a υa(COO) is observed at 1585 cm and υs(COO) at
-1
1
400 cm apart from the skeletal vibrations of the ligands. The separation between these
-1
two frequencies (ꢀυ = 185 cm ) adequately supports bidentate coordination of the acetato
7

group [8]. The broad band
υ(O-H) water is absent in the spectrum of the ligand (HL ), but is
n
-1
present in the complexes at 3185-3360 cm [8,26] (Fig. S3)
.
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The IR spectra of the ligands (HL ) exhibit a broad medium intensity band ranging
n
-1
-1
from 3100-3500 cm with peak centered at ~3370±12 cm which can be assigned to the
stretching vibration of phenolic (O-H) group [8]. The disappearance of the (O-H) phenolic
(C-O) vibration of C-
υ
-1
and a medium intensity bands at 1260-1220 cm can be attributed to
υ
O-H group [7,27], supports the contribution that proton displacement from the phenolic –
OH groups through the metal ion forming a bond link between the oxygen of the phenolic
-1
group and the metal ion. A band appearing at 1466 cm assigned to
υ
(N=N) is shifted to
-1
lower frequency on complex formation by 8-17 cm , showing that the coordination through
-1
one of the azo nitrogen atom [8]. New bands in all complexes at 580-624 and 407-470 cm
are assigned to (Cu-O) and (Cu-N), respectively. The (NH ) in all ligands under
investigation still lie at the same position in the spectra of the complexes.
υ
υ
υ
2
3.2. Mass spectra
The electron impact mass spectra of ligand (HL ) and its complex (3) are recorded and
3
investigated at 70 eV of electron energy. It is obvious that, the molecular ion peaks are in
good agreement with their suggested empirical formula as indicated from elemental analyses
(
Table 1). The mass spectra fragmentation mode of the ligand HL shows the exact mass
3
of ligand is 247.5 corresponding to the formula C H N OCl and mass of its complex
12
10
3
3
87.28 corresponding to the formula CuC H N O Cl as shown in Fig. S4(a and b),
14 14 3 4
respectively. The ion of ligand m/z = 247.5 undergoes fragmentation to a stable peak at m/z
219.5 by losing N atoms (structure ) as shown in Fig. 3. The loss of Cl leads to the
=
I
2
fragmentation with m/z = 184 (structure II). The loss of NH atoms leads to the
2
fragmentation with m/z = 168 (structure III). A breakdown of the backbone of HL ligand
3
gives the fragment (IV). The mass spectrum fragmentation modes of complex (3) shows the
m/z=387.046 and undergoes fragmentation to stable at m/z =214.5 by losing H O, Cu, OAc
2
and ONH (structure I) as shown in Fig. 4. The loss of N leads to fragmentation with m/z =
2
2
1
86.5 (structure II). A breakdown of the backbone of complex gives the fragments (III).
3
.3. X-ray diffraction
The X-ray diffraction (XRD) patterns for the powder form of complexes
[
Cu(L )(OAc)]H O (1) and [Cu(L )(OAc)]H O (4) are presented in Fig. 5. The XRD show
1 2 4 2
many diffraction peaks which confirm the polycrystalline phase while the complex
[
Cu(L )(OAc)] H O (2) is amorphous/polycrystalline mixture phase. The average crystallite
2 2
size (
ξ
) and The dislocation density (
δ
) can be calculated from the XRD pattern according to
8
following equation [23]:

K
λ
ξ
=
,
β1/ 2 cos
θ
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1
2
δ =
ξ
where λ is wavelength of X-ray radiation (1.540598 Å), K is a constant taken as 0.95 for
organic compounds and β1/2 is full width at half maximum of the reference diffraction peak
measured in radians and θ is the angle of diffraction. The calculated values of ξ are found
241 and 270 nm for complex (1) and complex (4), respectively. The calculated values of δ
-3
-5
-2
are found to be 1.72× 10 , 1.37× 10 nm for complexes (1) and (4), respectively. The
diffraction peaks in powder spectra are indexed and the lattice parameters are determined
with the aid of CRYSFIRE computer program [28]. The optimum indexed Miller indices,
(
hkl), for complexes under investigation are determined by using CHEKCELL program [29]
and presented in Fig. 5. The calculated lattice parameters which are a, b, c, and in
addition to crystal system and space group are presented in Table 2
α,
β
γ
.
3.4. Thermal analyses
Thermal analysis by the TGA techniques has proved to be very useful in determining
the crystal water content in complexes and the percentage of loss of masses of the ligands
and their thermal stability. The experimental results showed that the thermal decomposition
of ligands (HL ) and their Cu (II) complexes included two and three main steps,
n
respectively. The determined temperature ranges and the corresponding percent mass losses
are given in Table S1 (Figs. S5 and S6).
HL ligand shows two decomposition steps, the first stage occur in the temperature
1
o
range 30-342 C is attributed to loss of a part of the ligand (C H N ) (Found 50.31 % and
7
7
2
o
Calc. 52.42%). The second stage in the temperature range 342-806 C loss of a part of the
ligand C H NO (Found 49.69% and Calc. 47.57%). The determined temperature ranges and
6
6
the corresponding percent mass losses are given in Table S1
.
All Cu (II) complexes (1-4) showed TG curves in the temperature range ~30-400 °C
loss of outer H O molecule followed by loss of CH COO molecule. The third stage is
2
3
related to loss of the part of ligand. The final weight losses are due to the decomposition of
the rest of the ligand and metal oxides residue (Table 3).
3.5. Thermodynamic parameters for thermal degradations
The thermodynamic activation parameters of decomposition processes of the ligands
(
HL , HL , HL and HL ) and their Cu (II) complexes (1, 2, 3 and 4) namely activation
1 2 3 4
*
*
energy (E ), enthalpy (
ꢀ
H ), entropy (
ꢀ
S ), and Gibbs free energy change of the
a
9

*
decomposition (ꢀG ) are evaluated graphically by employing the Coast-Redfern [30] and
Horowitz-Metzger [31] methods and drown of Coats–Redfern (CR) and Horowitz-Metzger
ACCEPTED MANUSCRIPT
(
HM) of the ligands (HL ) and their Cu(II) complexes (1-4) (Figs. 6 and 7), respectively.
n
The calculated values of E , A,
ꢀ
S*,
and complexes are summarized in Table 4. According to the kinetic data obtained from
TGA curves, the negative values of activation entropies S* indicate more ordered activated
ꢀH* and ꢀG* for the decomposition steps for ligands
a
ꢀ
complexes than the reactants. It was found that the thermal stability of the metal complexes
was higher than that of the ligands.
3.6. Molecular structure
Molecular structures for ligands (HL ) and their complexes are shown in Fig. 8
.
n
Molecular structures (HOMO & LUMO) are presented in Fig. S7. Selected geometric
parameter bond lengths and bond angles are tabulated in Tables S2-S5 and 5-8 for the
ligands HL and their complexes, respectively. The HOMO–LUMO energy gap (ꢀE) is an
n
important stability index which is applied to develop theoretical models for explaining the
structure and conformation barriers in many molecular systems. The calculated quantum
chemical parameters are given in Table 9, global electrophilicity,
additional electronic charge, and Nmax, have been calculated [8]. It seems that the
electronegativity ( ) of complexes higher than ligands and electronegativity ( ) of ligands
ω, global softness, S and
ꢀ
χ
χ
increase from HL to HL4 and also electronegativity of complexes increased. This can be
1
attributed to the fact that the effective charge increased due to the electron withdrawing p-
substituent (HL and HL ) while it decreased by the electrons donating character of (HL ).
3
4
1
3.7. Molecular docking
Breast cancer rates gradually increased since the 1940s in many industrialized
countries. HER2 is a member of the epidermal growth factor receptor (EGFR / ERBB)
family. Amplification of this oncogene has been shown to play an important role in the
development and progression of some aggressive types of breast cancer. In recent years, the
protein biomarker becomes important goal of treatment for about 30% of breast cancer
patients [32].
Results of docked ligands were analyzed with the 2a91 as shown in Fig. S8 (A and C).
The results showed a possible arrangement between ligands (HL ) and receptor (2a91). The
n
docking study showed a favorable interaction between ligands HL and the receptor 2a91
n
and the calculated energy is listed in Table S6 and Fig. S8 (A and C) for receptor 2a91,
respectively. According to the results obtained in this study, HB plot curve indicate that, the
ligands HLn binds to the protein with hydrogen bond interactions and decomposed
interaction energies in Kcal/mole were exist between the of ligands HL with (2a91)
n
receptor as shown in Fig. 9 and Table S7. The ligands (HL ) have a great affinity (pk ) for
n
I
10

(2a91) receptor as shown in Table S6. The calculated efficiency is favorable where Ki
values estimated by AutoDock were compared with experimental K values, when available,
I
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and the Gibbs free energy is negative. Also, based on this data, we can propose that
interaction between the (2a91) receptor and the ligands (HL ) is possible. 2D plot curves of
n
docking with ligands (HL ) are shown in Fig. S9. Binding energies are most widely used as
n
mode of measuring binding affinity of a ligands. Thus, decrease in binding energy due to
mutation will increase the binding affinity of the azodye ligands toward the receptor. As the
electron density on azo dye group decrease the bending of our ligands with the receptor
(
2a91) increase, (HL4 ˃ HL3˃ HL2 ˃ HL ). The characteristic feature of azo dye ligands was
1
represented in presence of several active sites available for hydrogen bonding. This feature
gives them the ability to be good binding inhibitors to the protein and will help to produce
augmented inhibitory compounds. The results confirmed also that, the azo dye ligands
derived from 3-aminophenol is efficient inhibitor of 2a91.
3
.8. Magnetic measurement and electronic spectra
The room temperature magnetic moment values (µ ) per copper ion for the complexes
eff
were 0.93-1.99 BM range which may suggest a square planar structure [33]. The observed
magnetic moments of the complexes are consistent with the presence of single unpaired
electron [34,35]. The electronic absorption spectra of the ligands and its copper complexes
were measured at room temperature in DMF solutions. The electronic absorption spectra of
-1
the ligands display one electronic absorption in the ranges 22222-21460 cm , which are
*
*
assigned to n→π and intraligand π→π transitions, respectively. The electronic spectra of
the copper (II) complexes display two electronic absorption bands in the ranges 17915-
-1
2
2
2
2
1
7430 and 19300-20410 cm , assignable to the transition B1g→ A1g and B1g→ Eg
indicating the possibility of a square planar complexes. These spectral features support the
square planar geometry around copper (II) center [33]. Additional bands are present in the
high-energy region are assignable to (O
36]. These spectral features are in agreement with the other square planar copper(II)
complexes [37].
.9. ESR spectra
→ Cu) and (N=N → Cu) charge-transfer transitions
[
3
To obtain further information about the stereochemistry and the site of the metal
ligand bonding and to determine the magnetic interaction in the metal complexes. ESR
spectral assignments of the Cu (II) complexes along with the spin Hamiltonian and orbital
reduction parameters are summarized in Table 10. The ESR spectra of the complexes (1-3)
have been studied to provide information about the hyperfine and super hyperfine structures
to elucidate the geometry and the degree of covalence of the metal-ligand bond. The spectra
of the complexes (1-3) show typical axial behavior with slightly different g and g values.
ll
l
11

The various Hamiltonian parameters have been calculated for these complexes (1-3)
(Table 10). The ESR spectra of copper(II) complexes exhibits axially symmetric g-tensor
ACCEPTED MANUSCRIPT
parameters with g > g > 2.0023 indicating that the copper site has a d ground state,
x2-y2
ll
⊥
characteristic of square planar geometry and axially symmetric [38]. It has been reported
that g value of copper(II) complexes can be used as a measure of the covalent character of
ll
the metal-ligand bond. If the value is more than 2.3, the metal-ligand bond is essentially
ionic and the value less than 2.3 is indicative of covalent character [39]. The present ESR
results show that g is lower than 2.3 in this case suggesting that the copper (II) complexes
ll
are covalent in nature [40]. Also the trend g > g > g observed for these complexes
ll
⊥
av
indicate that the unpaired electron is most likely in the dx2-y2 orbital of the copper(II) ion and
are characteristic for the axial symmetry [41]. In axial symmetry, the g-values are related to
the G-factor by the expression G = (g -2)/(g -2) = 4. According to Hathaway and Billing
ll
⊥
[
41], if the value of G is greater than 4, the exchange interaction between copper (II) centers
in the solid is negligible, whereas when it is less than 4, a considerable exchange interaction
exists in the solid complex. The calculated G values (Table 10) lie within the range 3.90 –
5.44 for the copper (II) complexes (1-3). This support the absence of exchange coupling
between copper (II) centers in the solid state [42]. The f is taken as in indication for the
stereochemistry of the copper (II) complexes. Addition has suggested this ratio may be an
empirical indication of the tetrahedral distortion of the square planar geometry. The values
-1
-1
lower than 135 cm are observed for square planar structures and those higher than 150 cm
for tetrahedral distortion complexes. The values for the complexes under investigation,
Table 10, showed that all complexes associated with a square planar ligand field around the
copper(II) centers.
2
Molecular orbital coefficient,
α
(a measure of the covalence of the in-plan σ-
2
bonding between a copper 3d orbital and the orbitals) and
β
(covalent in-plan
π-bonding)
was calculated [43-48], which may be regarded as measure of covalence of the in-plane σ-
2
bonding, in-plane
π-bonding and our of plane
π-bonding, respectively. The value of
α
= 1.0
2
indicates complete ionic character, whereas
α
= 0.5 denotes 100% covalent bonding, with
the assumption of negligible small values of the overlap integral. According to Hathaway
2
2
2
[
49,50], the calculated values of K , K and K , Table 11, showed that Kll
≈
K⊥
≈ 0.77 for
ll
⊥
pure in-plane
σ
-bonding and K < K for in-plane
π-bonding, while for out of plane π-
ll
⊥
bonding K > K . In all the Cu(II) complexes, it is observed that K < K which indicates the
ll
⊥
ll
⊥
2
2
presence of significant in-plane
π
-bonding. The values of bonding parameters
-bonding and in plane -bonding. It was found that the
, g and G of Cu(II) complexes (1-3) g , A ×10 cm and β values are
α and β <
1.0 indicate significant in-plane
σ
π
2
2
-4
-1
2
values of
β
,
α
ll
ll
ll
12

dependent of the substituents effect of the p-position of the ligand and can be order as: p-(Cl
H > CH ) as shown in Fig. 10
>
.
3
ACCEPTED MANUSCRIPT
3
.10. Catalytic oxidation of alcohols
Copper (II) complexes have been well investigated in catalyzing the selective
oxidation of alcohols to aldehydes in the past decade [51,52]. Compared to the other transition
metal compounds, copper catalysts are commercially inexpensive, easily to be prepared and
handled. Although some Cu(I)/Cu(II) salts and their composites [53] have shown good
catalytic activity towards such reactions, the coordination compounds of copper with various
ligands are mostly investigated because of the solubility, the steric hindrance, the stability and
the redox properties of these catalysts could be easily adjusted by the ligands used [54-56].
However, catalytic methods based on transition metals need stringent reaction
conditions and high cost of the ligand, or have the disadvantages like long reaction time and
high temperature. Therefore, the development of practical, inexpensive, simple and green
chemical process for oxidation is still needed. We are interested in the use of hydrogen
peroxide, since it is cheap and sufficiently environment-friendly to be used on a commercial
scale. Herein, we investigate the catalytic oxidation of benzyl alcohol by the catalyst system,
[
Cu(L )(OAc)]H O/H O at room temperature. As a typical reaction procedure, oxidation
1 2 2 2
experiments have been performed as follows: 10 mmol of 30% H O was added drop wise to
2
2
the solution of the complex (0.01 mmol) and alcohol (2 mmol) in a mixture of DMF and H O
2
as solvents and the reaction mixture irradiated ultrasonically for 0.5 h. The color of the reaction
mixture changes from brown to green; this is probably due to coordination of the alcohol to the
copper (II) ion.
-1
However, oxidation of benzyl alcohol in 80% with turnover frequency (TOF) 10.6 h .
The catalytic oxidation of benzyl alcohol as a model substrate has been performed in the
absence of [Cu(L )(OAc)] H O and in the presence of some other co-oxidants like NaIO ,
1
2
4
K S O , NaBrO and NaOCl instead of H O and gave very low yield of benzaldehyde (less
2
2
8
3
2
2
than 5%). This is probably due to the formation of the solid precipitates NaIO , K SO , NaBr
3
2
4
and NaCl, respectively, which remain at the end of the reaction and make the workup, is
difficult. We conclude that the use of [Cu(L )(OAc)]H O is preferred catalyst for oxidation of
1
2
benzyl alcohol to benzaldehyde. The mechanism of this reaction could be explained on the
basis of the catalytic cycle shown below. Hydrogen peroxide oxidizes the produced Cu(I) ion
to Cu(II) ion again which in turn oxidizes the remaining benzyl alcohol. This catalytic cycle
continue till all the substrate is completely consumed. The novelty in this technique towards
this complex has not been tested before for this catalytic oxidation reaction in addition, this
catalytic system is good for catalytic oxidation of these compound since (1) it works at room
temperature, (2) the yield is comparable to other catalytic systems, (3) the work-up is simple
and (4) reaction time is short.
13

Cu(I)
H O
2
2
Benzaldehyde
ACCEPTED MANUSCRIPT
Co-oxidant Catalyst
Substrate
Benzyl alcohol
H O
2
Cu(II)
3.11. Biological studies
3.11.1. ABTS antioxidant activity
The antioxidant activities of azo ligands and its metal complexes were evaluated by in vitro
methods in order to compare the results. The evaluation study was carried out with different
concentrations of ligand and metal complexes and ascorbic acid was used as a standard.
By comparing the results obtained of antioxidant of the azo ligand compounds reported
in this study compounds HL , HL , HL and complex (1) showed high antioxidant activity in
1
2
3
the sequence HL > HL > HL > HL according to decreasing of electronegativity on azo
1
2
3
4
group (Table 11). Also we found that copper complex of ligand HL is the only complex which
1
increase the antioxidant activity than its ligand.
3.11.2. Antibacterial activity
The diseases caused by microbial infections were a serious menace to the health of
human being and often have connection to some other diseases when the body system gets
debilitated. Developing antimicrobial drugs and maintaining their potency in opposition to
resistance by different types of microorganisms as well as a broad spectrum of antibacterial and
antifungal activities were of major concerns of research in this area.
Antimicrobial activity of the synthesized compounds were screened in vitro against
gram positive and gram negative bacterial and a yeast in comparison with control drugs
ampicillin as an antibacterial agent and colitrimazole as an antifungal agent, by the agar
diffusion technique [57,58]. Biological evaluation in vitro revealed that compounds HL , HL ,
1
2
HL and complex (1) exhibited moderate to slight inhibitory action activity against
3
Staphylococcus aureus and Escherichia coli. The rest of compounds showed slight to no
sensitivity at all to the tested microorganisms. Complex (1) showed good activity against E.
coli with zones of inhibition 13 mm and activity index of 54.2% and compound HL also
2
showed moderate activity with zone 16 mm and activity index of 72.7 % (Table 12).
Compounds HL , HL , HL and complex (1) exhibited antifungal activity against C.
1
2
3
albicans with inhibition zones of 4,11,13 and 7 mm and activity index of 15.4%,42.3% 50%
and 26.9%, respectively. By investigating the variation in the antimicrobial activities of the
tested compounds it was revealed that the presence of Cl, NO in some compounds such as HL3
2
14

and HL may be result moderate activities against tested microorganisms when compared with
4
standard compounds.
ACCEPTED MANUSCRIPT
3.11.3. Cytotoxic
In view of the biological activity of azo dye complexes, we firstly evaluated the ability
of 3-aminophenol derivatives to inhibit cancer cell growth against Hepatocellular carcinoma
liver (HCC). In our experiments, half maximal inhibitory concentration (IC ) values
5
0
(compound concentration that produces 50% of cell death) were calculated. For comparison
purposes the cytotoxicity of Fluorouracil (5-FU) and the free ligand as well as the metal
complexes was evaluated under the same experimental conditions. The IC50 of the
complexes are higher than that of ligands, and there is no synergist effect with the
complexation with the metal. A slight decrease of IC is observed only for the [Cu(L )
5
0
1
(
OAc)]H O complex against MCF-7. Therefore, the chelation of the free ligand with metal
2
ion is essential for anticancer activities of these complexes. Importantly, it should be
emphasized that Cu(II) complexes exhibits considerable cell growth inhibition activity
against human liver hepatocellular carcinoma HepG2 cells similar to that of 5-FU (7.9 lg/ml)
for HePG2. These gratifying results are encouraging its further screening in vitro. Therefore,
its further biological evaluation in vitro as well as studies of mechanism of action is
necessary.
Cytotoxic and antitumor activity of ligands (HL ) and its metal complexes were tested
n
against (HePG2) and MCF-7cell line was detected by using different concentrations of the
tested compounds (1.56, 3.125, 6.25,12.5, 25,50 1.56 µg/ml) and viability cells (%) were
determined by the colorimetric method [59,60].
The results revealed that all tested compounds have cytotoxic and antitumor activity
against the breast carcinoma cell line. HL ,HL ligands and complex (1) were found to
2
3
exhibit potent anticancer activity which have better activity than complex (4). In Table 13
values of IC decrease activity of compounds increased against the tumor and we can apply
5
0
as anti-tumors drugs against tumors. The results shown in Fig. S10 and Fig. 11 revealed
that all tested compounds have cytotoxic and antitumor activity against the breast carcinoma
cell line and human liver hepatocellular carcinoma with superiority.
3.12. DNA binding studies
Absorption titration studies is one of the most universally methods used to study the
binding modes linking DNA compounds [61,62]. The phenolic -OH group of the ligands
may enhance their affinity towards DNA binding through formation of hydrogen bonding.
Absorption titration experiments were performed with fixed concentrations of the ligands
HL and Cu(II) complexes (
1
-4
) (40
ꢁM) while gradually increasing concentration of DNA
n
in 25 °C at 442, 488, 439 and 453 nm for the ligands (HL ), respectively and 466, 453, 454
n
15

and 456 nm for Cu(II) complexes (1-4), respectively. The absorption spectra of these ligands
and Cu(II) complexes with increasing concentration of CT-DNA in the range 300-600 nm
ACCEPTED MANUSCRIPT
are shown in Fig. S11 and Fig. 12, respectively.
Upon the addition of increasing amount of CT-DNA, a significant hyperchromic
effect was observed accompanied by a moderate red shift of 2–3 nm, indicative of
stabilization of the DNA helix. These spectral characteristic suggest that the ligands and
complexes bind either to the external contact electrostatic binding or to the major and minor
grooves of DNA. Moreover, this hyperchromic effect can be explained on the basis of these
two phenomena. The intrinsic binding constants (K ) of all the ligands (HL ) and Cu(II)
b
n
complexes (
1
-4
) with CT-DNA were determined using Eq. 2 [63].
The values of K obtained from the absorption spectral technique for ligands (HL )
b
n
5
5
5
5
-1
were calculated as 2.68×10 , 3.05 ×10 , 3.52 ×10 and 4.62 ×10 M , respectively.
The K values obtained from the absorption spectral technique for Cu(II) complexes (1-4)
b
5
5
5
5
-1
were calculated as 3.06 ×10 , 4.85×10 , 5.97×10 and 8.62 x10 M , respectively. The
binding constant of the complexes ( ) are comparatively higher than that of the ligands
HL ) due to formats six ring. The higher values of the binding constant of the ligands HL4
1-4
(
n
and HL are due to the presence of electron withdrawing group NO and Cl, respectively, as
3
2
shown in Fig. 13
.
Conclusions
The structures of Cu (II) complexes of the ligands (HL ) were confirmed by elemental
n
1
analyses, IR, H- NMR, X-ray, molar conductance, mass spectra and thermal analysis data.
It was found that the electron density on azo dye group decrease the bending of our ligands
with the receptor (2a91) increase, (HL4
˃ HL3˃ HL2˃ HL ) and the results confirmed also
1
that, the azo dye ligands derived from 3-aminophenol is efficient inhibitor of 2a91. The
comparison of cytotoxicity indicated that the complexes (1) and (3) showed much lower IC50
values for HePG2 with cell line and also complexes (1) and (2) lower IC values for MCF-
5
0
7
as well as HL , HL ligands and complex (1) were found to exhibit potent anticancer
2 3
activity which have better activity than complex (4). By comparing the results obtained of
ABTS antioxidant activity of the azo ligand compounds reported in this study compounds
HL , HL , HL and complex (1) and found that copper complex of ligand HL is the only
1
2
3
1
complex which increase the antioxidant activity than its ligand and the binding constant of
the complexes ( ) are comparatively higher than that of the ligands (HL ) due to formats
six ring. The higher values of the binding constant of the ligands HL and HL are due to
1-4
n
4
3
the presence of electron withdrawing groups. Catalytic oxidation of alcohols to the
corresponding aldehydes using Cu complexes was studied. The synthesized compounds
have antibacterial activity against the Gram-positive bacteria: Staphylococcus aureus and
16

also against the Gram-negative bacteria: Escherichia coli and Candida albicans and this is
may be due to the presence of electron withdrawing groups (NO and Cl).
2
ACCEPTED MANUSCRIPT
Acknowledgement
The authors would like to thank Prof. Dr. M.I. Abou-Dobara, Botany Department, Faculty
of Science, Damietta University, Egypt for his help during testing antimicrobial activity.
Appendix A: Supplementary material
See the attached file.
17

References
[
1] S. Yin, H Xu, W. Shi, Y. Gao, Y. Song, J. W. Y. Lam and B. Z. Tang, J. Polym., 46,
7
670 (2005).
ACCEPTED MANUSCRIPT
[
2] Y. Nabeshima, A. Shishido, A. Kanazawa, T. Shiono, T. Ikeda and T. Hiyama, Chem.
Mater., 9, 1480 (1997).
[
[
3] T. Pooja and C. Malay, Indian J. Eng. Mater. Sci. , 11, 499 (2004).
4] T.M. Robert, N.B. Robert and S.K. Bhattacharjee, Organic Chem. 6th Ed, New Delhi,
Pearson Prentice Hall, 1096 (2011).
[
5] H. Zollinger;"Color Chemistry Syntheses, Properties, Application of Organic Dyes and
Pigments". 3rd Edition. Wiley-VCH (2003).
[
6] L. Savanni, L. Chiasserini, A. Gaeta and C. Pellerano, Bioorg. Med. Chem., 10, 2193
2002).
7] A.Z. El-Sonbati, A.A.M. Belal, M.S. El-Gharib, Sh.M. Morgan, Spectrochim. Acta A
5, 627 (2012).
(
[
9
[
[
[
8] A.Z. El-Sonbati, M.A. Diab, Sh.M. Morgan, J. Mol. Liq. 225, 195 (2017).
9] M. Badea, R. Olar, E. Cristurean, D. Marinescu, A. Emandi, P. Budrugeac and E.
Segal,. J. Therm. Anal. Calor., 77, 815 (2004).
10] S.O. Podunavac-Kuzmanovi
Periodica Techn., 33, 151 (2002).
11] S. O. Podunavac-Kuzmanovi and LJS. Vojinovi
2003).
12] A.Z. El-Sonbati, M.A. Diab, A.A.M. Belal, Sh.M. Morgan, Spectrochim. Acta A 99,
53 (2012).
ć, V.M. Leovac, G.S. Ćetkovi and S.L. Markov, Acta
[
ć
ć, Acta Periodica Tech., 34, 119
(
[
[
3
13] C.H. Ng, H.K.A. Ong, C.W. Kong, K.W. Tan, R. Rahman, B.M.Yamin, and S.W.Ng,
Polyhedron, 25, 3118 (2006).
[
[
14] M.J. Waring , J. Mol. Biol , 13, 269 (1965).
15] M.F. Reichmann, C.A. Rice, C.A. Thomos and P. Doty, J. Am. Chem. Soc., 76 , 3047
(1954).
[
[
16] A. Wolfe, G.H. Shimer and T. Meehan, Biochem., 26, 6392 (1987).
17] A.B.A. El-Gazar, A.M.S. Youssef, M.M. Youssef, A.A. Abu-Hashem and F.A. Badria.
Eur. J. Med. Chem., 44, 609 (2009).
[
18] H.J. Mauceri, N. N. Hanna, M.A. Beckett, D.H, Gorski, M. J Staba, K. A. Stellato,
K.Bigelow, R . Heimann, S . Gately, M. Dhanabal, G.A. Soff, V.P. Sukhatme, D.W
Kufe and R. R, Weichselbaum, Nature, 394, 287 (1998) .
[
[
19] Z. Bikadi and E. Hazai. J. Chem. Inf., 1, 15(2009).
20] T. A. H. Merck, J. Comp. Chem.,17, 490 (1998).
18

[
21] G. M. Morris and D. S. Goodsell, J. Comp. Chem., 19, 1639(1998).
ACCEPTED MANUSCRIPT
[
22] H.M. Refaat, H.A. El-Badway, Sh.M. Morgan, J. Mol. Liq. 220, 802 (2016).
[
[
23] N.A. El-Ghamaz, A.Z. El-Sonbati, Sh.M. Morgan, J. Mol. Struc. 1027, 92 (2012).
24] G.G. Mohamed, A.A. El-Sherif, M.A. Saad, S.E.A. El-Sawy, Sh.M. Morgan, J. Mol.
Liq. 223 (2016) 1311–1332.
[
25] M.I. Abou-Dobara, A.Z. El-Sonbati, Sh.M. Morgan, World J. Microbiol. Biotechnol.
2
9, 119 (2013).
26] A.Z. El-Sonbati, M.A. Diab, M.M. El-Halawany, N.E. Salam, Spectrochim. Acta A 77,
55 (2010).
[
7
[
27] K. Nakamoto, " Infrared and Raman Spectra of Inorganic and Coordination
Compounds: Applications in Coordination Organometallic and Bioinorganic
Chemistry " Part B, John Wiley and Sons, pp 59-62 (1997).
[
[
28] R. Shirley, The CRYSFIRE System for Automatic Powder Indexing: User’s Manual,
The Lattice Press, Guildford, Surrey GU2 7NL, England, (2000).
29] J. Laugier and B. Bochu, LMGP-suite suite of programs for the interpretation of X-ray
experiments, ENSP/Laboratoire des Materiaux et du Genie Physique, B P 46.38042,
Saint Martin d’Heres, France, (2000).
[
[
[
30] W. Coats and J.P. Redfern, Nature, 20, 68 (1964).
31] H.W. Horowitz and G. Metzger, Anal. Chem. 35, 1464 (1963).
32] Z . Mitri, T . Constantine and R . O'Regan, Chem. Res. Pra., 2012 (2012), Article ID
7
43193.
33] M.M. Ibrahim, A.M. Ramadan, G.A.M. Mersal and S.A. El-Shazly. J. Mol. Struct.,
98, 1 (2011).
[
9
[
[
34] A. Sreekanth and M. R. P Kurup. Polyhedron, 22 , 3321 (2003).
35] F. A. Cotton and G .Wilkinson. Advanced Inorganic Chemistry. New Delhi, India:
Wiley Eastern (Pvt) (1969).
[
[
[
36] K. Singh, M.S. Barawa and P. Tyagi, Eur. J. Med. Chem., 42, 394 (2007).
37] M. Gaber, S.A. El-Daly and Y.S.Y. El-Sayed. J. Mol. Struct., 922, 51 (2009).
38] F.A. Cotton and G. Wilkinson, "Advanced Inorganic Chemistry", A Comprehensive
Text. 4 th Ed., John Wiley and Sons, New York (1986).
[
[
[
39] B.J. Hathaway and D.E. Billing, Coord. Chem. Rev.,5, 81 (1970).
40] H. Montgomery and E.C. Lingefetter, Acta Cryst., 20, 628 (1966).
41] C.J. Carrano, C.M. Nunn, R. Quan, J.A. Bomadies and V.I. Pecoran, Inorg. Chem., 29,
9
44 (1990).
[
[
42] D. Kilveson, J. Chem. Phys., 8, 8631 (1997).
43] M. Salavati-Niasari and F. Davar, Inorg. Chem. Commun., 9 , 304 (2006).
19

[
44] J. Piera and J.E. Backvall, Angew. Chem. Int. Ed., 47, 3506 (2008).
[
45] T. Chattopadhyay, M. Kogiso, M. Asakawa, T. Shimizu and M. Aoyagi, Catal.
ACCEPTED MANUSCRIPT
Commun., 12 , 9 (2010).
[
[
[
[
[
46] D. Ramakrishna and B.R. Bhat, Inorg. Chem. Commun., 14, 690 (2011)
47] C. Parmeggiani and F. Cardona, Green Chem., 14, 547 (2012).
48] J.E. Steves and S.S. Stahl, J. Am. Chem. Soc. 135, 15742 (2013).
49] D. Ramakrishna and B.R. Bhat, Inorg. Chem. Commun.,14, 690 (2011).
50] A. Rezaeifard, M. Jafarpour, A. Naeimi and M. Salimi, Inorg. Chem. Commun., 15,
2
30 (2012).
51] M.F. Semmelhack, C.R. Schmid, D.A. Cortes and C.S. Chou, J. Am. Chem. Soc., 106,
374 (1984).
[
3
[
[
[
[
52] L. Liu, J.L. Yan and W.Y. Yang, Catal. Commun., 9, 1379 (2008).
53] B.Z. Zhan and A. Thompson, Tetrahedron , 6, 2917(2004).
54] G. Ragagnin, B. Betemeier, S. Quici and P. Knochel, Tetrahedron, 58., 3985 (2002).
55] P. Gamez, I.W.C.E. Arends, J. Reedijk and R.A. Sheldon, Chem. Commun., 2414
(2003).
[
(
(
56] (a) N. Jiang and A.J. Ragauskas, Org. Lett., 7, 3689 (2005) .
b) S. Striegler, Tetrahedron, 62 , 9109 (2006).
c) S. Mannam, S.K. Alamsetti and G. Sekar, Adv. Synth. Catal., 349, 2253 (2007).
[
57] A. K. Sadana, Y. Mirza, K. R. Aneja, and O. Prakash, Eur. J. Med. Chem., 38, 533
2003).
(
[
58] M.O. Agwara, P. T. Ndifon, N. B. Ndosiri, A. G. Paboudam, D. M. Yufanyi, and A.
Mohamadou, Bull. Chem. Soc. Ethiopia, 24, 383 (2010).
[
[
59] T. Mosmann, J. Immunol. Methods, 65, 55 (1983).
60] P. Vijayan, G. Raghu, G. Ashok, S.A. Dhanaraj and B. Suresh, Indian J. Med. Res.,
1
20, 24 (2004).
[
[
61] M. S. Donnenberg , T. S. Whittam and E. coli, J. Clin. Invest., 107 , 539 (2001).
62] L. Shivakumar, K. Shivaprasad and H. D. Revanasiddappa, Spectrochim. Acta A107,
2
03 (2013).
63] C. Icsel, V.T. Yilmaz, F. Ari, E.Ulukaya and W.T.A. Harrison, Eur. J. Med. Chem.
0386 (2013).
[
6
20

Table 1. Physical properties and elemental analysis data of Cu(II) complexes (1-4).
ACCEPTED MANUSCRIPT
Yield
%
Exp. (Calc.) %
o
Compound
M.p. C
C
H
N
4
8.98
3.89
(4.09)
3.43
(3.69)
2.89
(3.10)
2.88
11.23
(11.46)
11.63
(11.91)
10.65
(10.85)
13.69
[
Cu (L )(OAc)] H O (1)
70
235
1
2
(
(
(
(
49.11)
47.55
[
[
Cu (L )(OAc)] H O (2)
68
64
243
260
2
2
47.65)
43.23
43.41)
42.07
42.26)
Cu (L )(OAc)] H O (3)
3
2
84
>300
[
Cu (L )(OAc)] H O (4)
4 2
(3.02)
(14.09)
21

Table 2. Crystal data for complexes (1 and 4)
Complex
(1)
(4)
ACCEPTED MANUSCRIPT
Space group
Crystal System
p2
p21
triclinic
monoclinic
Unit-cell dimensions
2
0.2460
a Å
b Å
16.5670
11.1560
1
8.8100
5.1780
90.00
99.88
90.00
1
c Å
α °
8.4430
93.99
β °
γ °
98.22
89.43
22

Table 3. Thermal analysis data of Cu II) complexes
.
TG weight loss
a
Temperature range (°C)
Complex
ACCEPTED MANUS
%
CRIPT
Found (Calc.)
4.79 (4.91)
Assignments
Cu
30-140
Outer H O molecule
2
1
3
40-314
14-800
>
17.65 (16.09)
30.44 (32.46)
47.16 (46.79)
21.94 (21.84)
30.92 (29.78)
47.08 (48.64)
3.26 (4.65)
16.29 (15.24)
49.59 (49.74)
30.84 (30.61)
7.07 (4.52)
CH COO
3
1
2
3
C H N
2
7
7
800
0-424
424-800
800
0-109
C H N + CuO
6 6
Outer H O molecule + CH COO
2 3
C H N
6 5 2
C H N + CuO
6 6
3
>
3
1
3
Outer H O molecule
2
09-397
97-795
CH COO
3
C H N Cl
9
7
3
>795
C H + CuO
3 3
3
2
3
0-251
51-377
77-800
Outer H O molecule
2
20.68 (22.39)
CH CO+ NO
3
2
4
24.31 (22.64)
48.02 (50.69)
C H N
6
4
>800
C H N O + CuO
6 5 2
a
Numbers as given in Table 1.
23

Table 4. The thermodynamic activation parameters of decomposition processes of the
a
ligands and their Cu(II) complexes .
ACCEPTED MANUSCRIPT
Parameter
Temp. range
Correlation
coefficient
r)
o
Compound
HL1
( C)
Method
Ea
A
(s )
-∆S*
∆H*
∆G*
−
1
−1
-1
-1
−1
−1
(
(
kJ mol )
(J mol K ) (kJ mol ) (kJ mol )
5
-3
-3
2
8
8
1.4
9.9
CR
HM
8.02X107
136 x10
113 x10
77.1
85.6
1.47 x102
0.95338
0.92419
1
3
1
2
40-342
42-806
24-298
98-802
1.38x10
1.44 x10
-3
-3
2
2
1
36 x10
4.9 x10
CR
HM
40.8
46.8
266 x10
250 x10
33.7
39.7
2.59 ×102
0.99916
0.99932
2
-3
2.52 ×10
x1022 6
-3
2
CR
HM
81.42
91.3
2.24x10
7.94x10
1.27 x10
9.77 x10
78
87.3
1.40 x102
0.98318
0.96551
7
-4
1.35 x10
HL2
HL3
-3
-3
2
CR
HM
21.8
29.9
2.55 x10
104 x10
284 x10
15
23
2.49 x102
0.9999
0.99959
-3
-3
2.72 x10
2.47 x10
3
-3
-3
2
CR
HM
65.7
72.4
5.32x105
1.77 x10
1.34 x10
61.7
68.5
1.46 x102
0.99158
0.98151
8
6-312
12-803
19-172
9.91x10
1.32 x10
3
-3
-3
2
CR
HM
25.8
36.3
2.10x10
1.90 x10
2.63 x10
18.9
29.4
1.77 x102
0.99831
0.99289
3
1
-1
3 x10
2.48x10
12
2
2
CR
HM
111
131
1.98x10
5.52x10
1.23 x102
107
128
1.12 x102
0.99717
0.97945
14
3.45 x10
1.13 x10
HL4
4
2
2
CR
HM
77.1
86.7
8.68x106
1.55 x102
72.6
82.2
1.56 x102
0.999
0.99472
1
72-355
2.48x10
1.27 x10
1.51 x10
8
2
2
CR
HM
90.1
96.3
2.93x10
8.62 x102
86.4
92.6
1.25 x102
0.98087
0.97456
1
2
1
40-205
05-252
53-281
9
2.78 x10
6.75 x10
1.23 x10
1
1
8
2
2
CR
HM
195
195
2.21 x10
4.09 x10
1.02 x102
191
190
1.40 x102
0.9886
0.9946
18
1.07 x10
1.37 x10
3
2
2
CR
HM
58.3
66.3
4.88 x104
1.78 x102
54.2
62.2
1.42 x102
0.99724
0.98846
9.52 x10
1.54 x10
1.38 x10
2
3
3
2
2
CR
HM
75.5
85.5
7.37x10
1.77 x102
70.3
80.3
1.81 x102
0.98731
0.97902
2
1
2
1
2
82-424
10-237
37-795
49-251
51-306
4
8.87 x10
1.56 x10
1.78 x10
2
2
2
CR
HM
32.2
42.2
1.81 x102
2.45 x102
27.8
37.8
1.57 x102
0.99768
0.99126
6.94 x10
2.14 x10
1.51 x10
2
2
2
CR
HM
50.9
66.1
129 x10
2.52 x102
43.7
58.9
2.63 x102
0.99003
0.98077
2
2.44 x10
2.27 x10
2.57 x10
4
2
2
CR
HM
60.6
69.3
1.66 x105
1.68 x102
56.7
65.4
1.36 x102
0.98038
0.98361
4.08 x10
1.41 x10
1.32 x10
4
15
2
2
CR
HM
181
191
1.05 x10
2.41 x10
3.75 x10
176
187
1.55 x102
0.98253
0.98304
16
2
6.36 x 10
1.52 x10
a
Numbers as given in Table 1.
24

Table 5. Selected geometric parameters for complex (1)
.
Bond lengths (Å)
o
Bond angles ( )
ACCEPTED MANUSCRIPT
C(22)-H(37) 1.115
H(37)-C(22)-H(36) 102.932
H(37)-C(22)-H(35) 102.878
H(37)-C(22)-C(20) 103.558
H(36)-C(22)-H(35) 112.767
H(36)-C(22)-C(20) 116.185
H(35)-C(22)-C(20) 116.037
C(22)-C(20)-O(21) 166.695
C(22)-C(20)-O(19) 78.599
O(21)-C(20)-O(19) 88.097
H(34)-C(17)-H(33) 107.859
H(34)-C(17)-H(32) 108.471
H(34)-C(17)-C(14) 110.001
H(33)-C(17)-H(32) 107.168
H(33)-C(17)-C(14) 112.506
H(32)-C(17)-C(14) 110.692
H(30)-C(15)-C(16) 119.197
H(30)-C(15)-C(14) 119.854
C(16)-C(15)-C(14) 120.948
C(15)-C(14)-C(13) 117.732
C(15)-C(14)-C(17) 121.702
C(13)-C(14)-C(17) 120.566
H(29)-C(13)-C(14) 119.401
H(29)-C(13)-C(12) 119.441
C(14)-C(13)-C(12) 121.158
H(31)-C(16)-C(9) 120.364
H(31)-C(16)-C(15) 117.479
C(9)-C(16)-C(15) 122.156
H(28)-C(12)-C(13) 117.038
H(28)-C(12)-C(9) 121.005
C(13)-C(12)-C(9) 121.95
Cu(18)-O(21)-C(20) 109.607
Cu(18)-O(19)-C(20) 95.938
O(21)-Cu(18)-O(19) 66.357
O(21)-Cu(18)-N(10) 121.366
O(21)-Cu(18)-O(8) 118.765
O(19)-Cu(18)-N(10) 117.501
O(19)-Cu(18)-O(8) 115.706
N(10)-Cu(18)-O(8) 110.646
C(16)-C(9)-C(12) 116.034
C(16)-C(9)-N(10) 120.465
C(12)-C(9)-N(10) 123.47
Cu(18)-N(10)-N(11) 112.469
Cu(18)-N(10)-C(9) 115.689
N(11)-N(10)-C(9) 114.831
H(27)-N(7)-H(26) 119.653
H(27)-N(7)-C(6) 120.166
H(26)-N(7)-C(6) 120.182
N(7)-C(6)-C(1) 120.193
C(22)-H(36) 1.116
C(22)-H(35) 1.116
C(17)-H(34) 1.114
C(17)-H(33) 1.113
C(17)-H(32) 1.114
C(16)-H(31) 1.105
C(15)-H(30) 1.103
C(13)-H(29) 1.103
C(12)-H(28) 1.104
N(7)-H(27) 1.049
N(7)-H(26) 1.049
C(5)-H(25) 1.104
C(2)-H(24) 1.104
C(1)-H(23) 1.103
C(9)-C(16) 1.353
C(15)-C(16) 1.342
C(14)-C(15) 1.343
C(13)-C(14) 1.343
C(12)-C(13) 1.342
C(9)-C(12) 1.353
O(21)-Cu(18) 1.812
O(19)-Cu(18) 1.806
C(20)-C(22) 1.534
C(20)-O(21) 1.246
O(19)-C(20) 1.581
N(10)-Cu(18) 1.315
O(8)-Cu(18) 1.804
C(14)-C(17) 1.51
N(11)-C(3) 1.276
N(10)-N(11) 1.272
C(9)-N(10) 1.282
C(4)-O(8) 1.37
C(6)-N(7) 1.268
C(6)-C(1) 1.341
C(5)-C(6) 1.342
C(4)-C(5) 1.347
C(3)-C(4) 1.353
C(2)-C(3) 1.346
C(1)-C(2) 1.341
N(7)-C(6)-C(5) 120.387
C(1)-C(6)-C(5) 119.417
Cu(18)-O(8)-C(4) 105.712
25

H(25)-C(5)-C(6) 119.66
H(25)-C(5)-C(4) 118.323
C(6)-C(5)-C(4) 122.015
ACCEPTED MANUSCRIPT
C(3)-N(11)-N(10) 126.372
O(8)-C(4)-C(5) 119.785
O(8)-C(4)-C(3) 122.116
C(5)-C(4)-C(3) 118.072
N(11)-C(3)-C(4) 122.521
N(11)-C(3)-C(2) 117.543
C(4)-C(3)-C(2) 119.927
H(24)-C(2)-C(3) 120.366
H(24)-C(2)-C(1) 118.547
C(3)-C(2)-C(1) 121.084
H(23)-C(1)-C(6) 120.837
H(23)-C(1)-C(2) 119.697
C(6)-C(1)-C(2) 119.466
26

Table 6. Selected geometric parameters for complex (2)
.
ACCEPTED MANUSCRIPT
Bond lengths (Å)
o
Bond angles ( )
C(21)-H(34) 1.115
C(21)-H(33) 1.116
C(21)-H(32) 1.116
C(16)-H(31) 1.104
C(15)-H(30) 1.103
C(14)-H(29) 1.103
C(13)-H(28) 1.103
C(12)-H(27) 1.105
N(7)-H(26) 1.049
N(7)-H(25) 1.049
C(5)-H(24) 1.104
C(2)-H(23) 1.104
C(1)-H(22) 1.103
C(9)-C(16) 1.354
C(15)-C(16) 1.342
C(14)-C(15) 1.34
C(13)-C(14) 1.34
C(12)-C(13) 1.342
C(9)-C(12) 1.354
O(20)-Cu(17) 1.812
O(18)-Cu(17) 1.806
C(19)-C(21) 1.534
C(19)-O(20) 1.246
O(18)-C(19) 1.581
N(10)-Cu(17) 1.315
O(8)-Cu(17) 1.804
N(11)-C(3) 1.276
N(10)-N(11) 1.272
C(9)-N(10) 1.283
C(4)-O(8) 1.37
H(34)-C(21)-H(33) 102.884
H(34)-C(21)-H(32) 102.93
H(34)-C(21)-C(19) 103.568
H(33)-C(21)-H(32) 112.759
H(33)-C(21)-C(19) 116.054
H(32)-C(21)-C(19) 116.165
C(21)-C(19)-O(20) 166.692
C(21)-C(19)-O(18) 78.594
O(20)-C(19)-O(18) 88.098
H(30)-C(15)-C(16) 120.055
H(30)-C(15)-C(14) 119.71
C(16)-C(15)-C(14) 120.235
H(29)-C(14)-C(15) 120.42
H(29)-C(14)-C(13)
120.4
C(15)-C(14)-C(13) 119.178
H(28)-C(13)-C(14) 119.798
H(28)-C(13)-C(12) 120.092
C(14)-C(13)-C(12) 120.109
H(31)-C(16)-C(9) 121.038
H(31)-C(16)-C(15) 116.964
C(9)-C(16)-C(15) 121.992
H(27)-C(12)-C(13) 117.416
H(27)-C(12)-C(9) 120.458
C(13)-C(12)-C(9) 122.125
Cu(17)-O(20)-C(19) 109.608
Cu(17)-O(18)-C(19) 95.937
O(20)-Cu(17)-O(18) 66.357
O(20)-Cu(17)-N(10) 121.518
O(20)-Cu(17)-O(8) 118.968
O(18)-Cu(17)-N(10) 117.426
O(18)-Cu(17)-O(8) 115.497
N(10)-Cu(17)-O(8) 110.553
C(16)-C(9)-C(12) 116.339
C(16)-C(9)-N(10) 123.359
C(12)-C(9)-N(10) 120.27
Cu(17)-N(10)-N(11) 112.381
Cu(17)-N(10)-C(9) 115.713
N(11)-N(10)-C(9) 114.911
H(26)-N(7)-H(25) 119.644
H(26)-N(7)-C(6) 120.165
H(25)-N(7)-C(6) 120.191
N(7)-C(6)-C(1) 120.206
C(6)-N(7) 1.268
C(6)-C(1) 1.341
C(5)-C(6) 1.342
C(4)-C(5) 1.347
C(3)-C(4) 1.354
C(2)-C(3) 1.346
C(1)-C(2) 1.341
N(7)-C(6)-C(5) 120.37
C(1)-C(6)-C(5) 119.421
Cu(17)-O(8)-C(4) 105.642
H(24)-C(5)-C(6) 119.666
H(24)-C(5)-C(4) 118.319
C(6)-C(5)-C(4) 122.012
C(3)-N(11)-N(10) 126.329
O(8)-C(4)-C(5) 119.779
27

O(8)-C(4)-C(3) 122.12
C(5)-C(4)-C(3) 118.073
N(11)-C(3)-C(4) 122.503
ACCEPTED MANUSCRIPT
N(11)-C(3)-C(2) 117.562
C(4)-C(3)-C(2) 119.926
H(23)-C(2)-C(3) 120.365
H(23)-C(2)-C(1) 118.549
C(3)-C(2)-C(1) 121.083
H(22)-C(1)-C(6) 120.836
H(22)-C(1)-C(2) 119.698
C(6)-C(1)-C(2) 119.466
28

Table 7. Selected geometric parameters for complex (3)
.
ACCEPTED MANUSCRIPT
Bond lengths (Å)
o
Bond angles ( )
C(22)-H(34) 1.115
C(22)-H(33) 1.116
C(22)-H(32) 1.116
C(16)-H(31) 1.104
C(15)-H(30) 1.103
C(13)-H(29) 1.103
C(12)-H(28) 1.105
N(7)-H(27) 1.049
N(7)-H(26) 1.049
C(5)-H(25) 1.104
C(2)-H(24) 1.104
C(1)-H(23) 1.103
C(9)-C(16) 1.353
C(15)-C(16) 1.342
C(14)-C(15) 1.341
C(13)-C(14) 1.341
C(12)-C(13) 1.342
C(9)-C(12) 1.354
O(21)-Cu(18) 1.812
O(19)-Cu(18) 1.806
C(20)-C(22) 1.534
C(20)-O(21) 1.246
O(19)-C(20) 1.581
N(10)-Cu(18) 1.315
O(8)-Cu(18) 1.804
C(14)-Cl(17) 1.726
N(11)-C(3) 1.276
N(10)-N(11) 1.272
C(9)-N(10) 1.283
C(4)-O(8) 1.371
C(6)-N(7) 1.268
C(6)-C(1) 1.341
H(34)-C(22)-H(33) 102.89
H(34)-C(22)-H(32) 102.933
H(34)-C(22)-C(20) 103.571
H(33)-C(22)-H(32) 112.758
H(33)-C(22)-C(20) 116.067
H(32)-C(22)-C(20) 116.144
C(22)-C(20)-O(21) 166.69
C(22)-C(20)-O(19) 78.595
O(21)-C(20)-O(19) 88.095
H(30)-C(15)-C(16) 119.351
H(30)-C(15)-C(14) 120.549
C(16)-C(15)-C(14) 120.1
C(15)-C(14)-C(13) 119.331
C(15)-C(14)-Cl(17) 120.29
C(13)-C(14)-Cl(17) 120.378
H(29)-C(13)-C(14) 120.644
H(29)-C(13)-C(12) 119.416
C(14)-C(13)-C(12) 119.939
H(31)-C(16)-C(9) 120.956
H(31)-C(16)-C(15) 116.977
C(9)-C(16)-C(15) 122.061
H(28)-C(12)-C(13) 117.394
H(28)-C(12)-C(9) 120.377
C(13)-C(12)-C(9) 122.229
Cu(18)-O(21)-C(20) 109.609
Cu(18)-O(19)-C(20) 95.934
O(21)-Cu(18)-O(19) 66.353
O(21)-Cu(18)-N(10) 121.535
O(21)-Cu(18)-O(8) 119.302
O(19)-Cu(18)-N(10) 117.441
O(19)-Cu(18)-O(8) 115.25
N(10)-Cu(18)-O(8) 110.43
C(16)-C(9)-C(12) 116.318
C(16)-C(9)-N(10) 123.394
C(12)-C(9)-N(10) 120.256
Cu(18)-N(10)-N(11) 112.3
Cu(18)-N(10)-C(9) 115.66
N(11)-N(10)-C(9) 115.033
H(27)-N(7)-H(26) 119.643
H(27)-N(7)-C(6) 120.168
H(26)-N(7)-C(6) 120.188
N(7)-C(6)-C(1) 120.197
C(5)-C(6) 1.342
C(4)-C(5) 1.347
C(3)-C(4) 1.354
C(2)-C(3) 1.346
C(1)-C(2) 1.341
N(7)-C(6)-C(5) 120.376
C(1)-C(6)-C(5) 119.424
Cu(18)-O(8)-C(4) 105.557
H(25)-C(5)-C(6) 119.663
H(25)-C(5)-C(4) 118.32
C(6)-C(5)-C(4) 122.015
C(3)-N(11)-N(10) 126.247
O(8)-C(4)-C(5) 119.778
29