Synthesis and characterization for novel Cu(II)-thiazole complexes-dyes and their usage in dyeing cotton to be special bandage for cancerous wounds

Article abstract of DOI:10.1016/j.molstruc.2019.05.080

New series of Cu(II)-thiazole complexes were synthesized using variable p-substituted, N-aryl-2-oxo-2-(thiazol-2-ylamino)-acetohydrazonoyl cyanide. All new syntheses were elucidated by, analytical, spectral and conformational study. The binuclear feature

Full text of DOI:10.1016/j.molstruc.2019.05.080

Accepted Manuscript
Synthesis and characterization for novel Cu(II)-thiazole complexes-dyes and their
usage in dyeing cotton to be special bandage for cancerous wounds
Nashwa El-Metwaly, Ismail Althagafi, Abdalla M. Khedr, Jabir H. Al-Fahemi, Hanadi
A. Katouah, Aisha S. Hossan, Aisha Y. Al-Dawood, Gamil A. Al-Hazmi
PII:
S0022-2860(19)30650-7
DOI:
https://doi.org/10.1016/j.molstruc.2019.05.080
MOLSTR 26587
Reference:
To appear in: Journal of Molecular Structure
Received Date: 14 April 2019
Revised Date: 15 May 2019
Accepted Date: 19 May 2019
Please cite this article as: N. El-Metwaly, I. Althagafi, A.M. Khedr, J.H. Al-Fahemi, H.A. Katouah, A.S.
Hossan, A.Y. Al-Dawood, G.A. Al-Hazmi, Synthesis and characterization for novel Cu(II)-thiazole
complexes-dyes and their usage in dyeing cotton to be special bandage for cancerous wounds, Journal
of Molecular Structure (2019), doi: https://doi.org/10.1016/j.molstruc.2019.05.080.
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ACCEPTED MANUSCRIPT
Synthesis and characterization for novel Cu(II)-thiazole complexes-dyes and their
usage in dyeing cotton to be special bandage for cancerous wounds
Nashwa El-Metwaly1,2*, Ismail Althagafi1, Abdalla M. Khedr1,3, Jabir H. Al-Fahemi1, Hanadi A. Katouah1, Aisha S.
Hossan4, Aisha Y. Al-Dawood1, Gamil A. Al-Hazmi 4,5
1Chemistry Department, College of Applied Sciences, Umm Al-Qura University, Makkah, Saudi Arabia
2Chemistry Department Faculty of Science, Mansoura University, Mansoura, Egypt
3Chemistry Department, Faculty of Science, Tanta University, Tanta, Egypt
4Chemistry Department, Faculty of Science, King Khalid University, P.O. Box 9004, Abha, Saudi Arabia
5Chemistry Department, Faculty of Applied Sciences, Taiz University, P.O. Box 82, Taiz, Yemen
Abstract
New series of Cu(II)-thiazole complexes were synthesized using variable p-substituted, N-aryl-2-oxo-
2-(thiazol-2-ylamino)-acetohydrazonoyl cyanide. All new syntheses were elucidated by, analytical,
spectral and conformational study. The binuclear feature (2M:1L), was proposed for all complexes,
through mono-negative tetra-dentate chelation mode in octahedral or square-planer geometry. The
1
formulae of chosen compounds, were confirmed by, HNMR and mass spectral analysis. The ideal
distribution for atomic-skeletons, was performed utilizing Gaussian09 software, to confirm the
bonding mode. Also, substantial parameters were extracted from output-files (log &chk), beside others
calculated based on frontier energy gaps. The superiority of Cu(II) complexes, was predicated from
such conformational study. Also and by applying MOE module, the docking process was performed
for most syntheses against selected pathogen proteins (1miu, 4k9g and 5jm5), which have been tested
practically in application. The extracted docking-data, showed clear superiority and promising
efficiency for Cu(II) complexes as anticancer drugs compared to free derivatives. Traditional screening
was conducted over new complexes against three carcinoma cell lines (HCT-116, MCF-7 & HepG-2)
as well as healthy cell line (HSF). IC50 values showed considerable toxicity of Cu(II)-4e complex
versus HCT-116 cell line. The antitumor screening was conducted over cotton fabric after dyeing by
complexes, to test the degree of success to be used as a special bandage for cancerous wounds. The
most lighted observation was, the effect of released-pigmenting complex on colon cancer cell line,
while the absence of any effect on healthy cell. Also, the released-pigment, controlled the pH of
cancerous wound, which is significantly preferable.
Keywords: Cu(II)-thiazole complexes, bandage for cancerous wounds, CT-DNA, molecular Docking, DFT/B3LYP
*Corresponding author: Prof. Nashwa M. El-Metwaly, E- mail address: n_elmetwaly00@yahoo.com, Telephone No.:
+966538553123
1. Introduction
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For many years, the heterocyclic compounds containing O, N, and S, displayed magnificent
importance in medicinal and chemical applications[1,2]. They are widely applied as cosmetics,
perfumes, food additives, pharmaceuticals, optical brighteners, laser and fluorescent and dyes[3].
Thiazole derivatives have enhanced antimicrobial activities towards various pathogens including
Helicobacter pylori and Mycobacterium tuberculosis[4]. These thaisole compounds, have also good
anti-cholinesterase, anti-cancer, anti-analgesic, anti-inflammatory activities[5]. Transition metals have
incorporated with numerous biological processes that are animated to life processes[6]. Hence, they
can coordinate with N- or O-terminals from proteins in a assortment of models, and so, play a peppy
role biological system such as the utility and conformation of living macromolecules [7]. Schiff bases
involving imino moieties (>C=N–) and their complexes have been exceedingly reported to display a
distinction of motivating biological effectiveness such as anti-inflammatory, antimicrobial, antifungal,
antitumor, analgesic effects, anti-proliferative, and anticancer activities[8]. Transition metal ion
complexes, containing different bioactive ligands, are often more efficacious regarding pharmaceutical
and biological applications compared with the free ligands [9]. Additionally, Schiff bases containing
thiazole nucleus are expected to exhibit great biological activities especially as antitumors [10]. Metal
complexes as therapeutic antitumor agents have been widely applied following the success of cis-
platin. Recently, cis-platin has been identified as one of the most extensively applied antitumor drugs
over the world, elevated efficiency for ovarian and testicular cancers, in the treatment of bronchogenic,
oropharyngeal, bladder, cervical, and carcinoma species [11]. Different complexes based on platinum
ions are clinically applied in order to help the inducement of cancer cells death. Platinum compounds
have toxic effects, despite the positive effects of them in killing cancer cells [12]. We have synthesized
different other metal complexes and tested for their anticancer and other bio-activities [13,14], to
overcome the disadvantages of platinum complexes. Furthermore, Cu(II) complexes derived from
Schiff bases have been reported as a promising potential chemotherapeutic agents [15]. From all
previously reported, especially that interested in Cu(II) complexes, they were fully succeeded in
biological applications concerning various pathways. From such point, and now we interested in
synthesis of new thiazole dyes, which utilized to synthesize their corresponding Cu(II) complexes.
After completing synthesis, all verification tools (analytical, spectral and conformational theoretical)
were implemented to extract full characterization bases. Furthermore and concerning the application
part, we conduct test to enhance anti-cancer impact by create innovative design for special bandage
that gains three prominent features. Such features as, close the wounds, eliminate bacteria from cancer
mass, which allowing chemotherapy to reach their target. Moreover, keep cancer cells under stable
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concentration of compound-released from bandage under the acidic conditions which already created
by cancer cells themselves.
2. Experimental part
2.1. Chemicals used
The reagents used to prepare diazonium salts as, NaNO2, HCL, aniline, 4-toluidine, 4-anisidine, 4-
acetoaniline and 4-chloroaniline, which were purchased from Sigma & Aldrich. Thiazolyl-
cyanoacetamide, which used for diazo-coupling, was PDH. CuCl2. 2H2O is a Fluka product which used
in complexation. While, the chemical of antitumor study were, SulphoRhodamine-B (SRB), were
purchased from Sigma Chemical Co. (St. Louis, MO, USA). Culture media and growth supplements
were purchased from Gibco / Life Technologies Co, (Carlsbad, CA, USA). Cell culture vessels were
purchased from Nunc Co. (Roskilde, Denmark). All solvents as, ethanol, methanol, DMF, DMSO were
PDH and used without previous treatments.
2.2. Synthesis
2.2.1. Synthesis of N-aryl-2-oxo-2-(thiazol-2-ylamino)-acetohydrazonoyl cyanide derivatives 4a-e
The preliminary step was the synthesis of diazonium salt, in which an aqueous solution of 0.21g
sodium nitrite (in 10mL H2O) was added gradually to cold solution (0 ) of the aromatic amine (0.003
mol) (namely; aniline, 4-toluidine, 4-anisidine, 4-acetoaniline and 4-chloroaniline in 3 mL conc. HCl.
Secondly, a diazocoupling process was executed after the addition of each diazonium salt solution to to
a cold solution (0 ) of thiazolyl-cyanoacetamide derivative 3 (0.003 mol, 0.50 g) in pyridine (30 mL),
with constant stirring till 2h (Scheme 1). The solid that obtained upon dilution with 10 mL cold water
was filtered off. Recrystallization process was carried out from EtOH-DMF (1:1) mixture, to pick up
acetohydrazonoyl cyanide dyes 4a-e[16]. The colored derivatives 4a-e were yielded by 67-76% and
their melting points are ranges of, 248-250, 228-230, 208-210, 268-270 and 252-254 , respectively.
The elements content and IR spectral study proposed the formulae of dyes 4a-e (Fig. 1S), which
suggested to found in tautomeric forms (keto/enol). 1H & 13C NMR spectral analysis (Fig. 1&2S) were
accomplished for two selected derivatives (4b &4c, in d6-DMSO), which asserted on formulae
1
suggested. The H NMR spectrum of 4b, H-TTAC derivative displayed signals at; δ = 2.31 (s, 3H,
CH3), 7.20 (d, J = 8.4 Hz, 2H, Ar-H), 7.28 (d, J = 3.6 Hz, 1H, thiazole-H5), 7.55 (d, J = 3.6 Hz, 1H,
13
thiazole-H4), 7.74 (d, J = 8.4 Hz, 2H, Ar-H), 12.05 (s, 1H, NH or OH), 12.39 (s, 1H, NH). ). Its C
NMR spectrum displayed signals at; δ = 21.60, 108.44, 112.73, 117.13 (2C), 124.73, 130.72 (2C),
131.76, 140.67, 154.46, 160.44, 163.46 ppm. While, the 1H NMR spectrum of 4c, H-ATAC displayed
signals at; δ =3.78 (s, 3H, OCH3), 7.00 (d, J = 8.4 Hz, 2H, Ar-H), 7.28 (d, J = 3.6 Hz, 1H, thiazole-
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H5), 7.55 (d, J = 3.6 Hz, 1H, thiazole-H4), 7.80 (d, J = 8.4 Hz, 2H, Ar-H), 12.31 (s, 1H, NH or OH).
Its 13C NMR spectrum displayed signals at δ = 55.92, 107.28, 111.72, 115.29 (2C), 118.88 (2C),
126.72, 129.81, 136.17, 152.91, 160.90, 164.54 ppm.
Scheme. 1.
2.2.2. Synthesis of Cu(II)-thiazole complexes
The complexes were synthesized by using equi-molar addition of CuCl2. 2H2O (2mmol, 0.341g) to
each corresponding thiazole derivative 4a-e in ethanolic solution. The reaction mixtures were heated
under reflux from 3-5h, the solid complexes were precipitated. All hot solutions were filtered off,
washed by ethanol and followed by diethyl ether. The complexes were dried in vacuum desiccators
under CaCl2. All complexes are stable, insoluble in common organic solvents but completely soluble in
DMSO and DMF, also having high m.p. over 300 oC.
2.2.3. Cotton-bandage pigmentation
0.2g from each prepared complex was dissolved in few drops of DMF and completed to 20 mL by
ethanol, then the dye bath was adjusted to pH=6 and heated till 90oC, after that, 1g from cotton fabric
was immersed. The dyeing solution was stirred over 60 min, after that cool the dye bath. Followed by
washing pigmented cotton by distilled water and in aqueous solution containing 1 g/L detergent for 10
min at 900C by a ratio of 20:1, then the dyed cotton-pieces are ready for application [17].
2.3. DNA binding
The degree of binding efficiency for new triazole derivatives towered CT-DNA has been investigated
using spectrophotometric method. 50 mg of CT-DNA were dissolved overnight in bi-distilled water
o
(pH= 7.0) through stirring and hold at 4 C. A buffering solution (Tris - HCl buffer) was prepared by
mixing solutions of 5.0 mM tris(hydroxymethyl)-aminomethane and 50 mM NaCl (in bi-distilled
water, pH= 7.0). A definite concentration from CT-DNA was prepared in Tris-HCl buffer. Such, gave
absorption at 260 and 280 nm by ratio; A260/A280 equal to 1.8-1.9 range, which is attributing to DNA
free from protein [18]. A known molar absorpitivity coefficient (6600 M−1cm−1) for DNA at 260 nm,
was used to estimate its concentration spectrophotometricaly (5.25×10−4 M). 200- 900 nm was the
scanning range for measurements using 1cm quartz cuvette at 25°C. A fixed concentration from each
compound (6a-6e) was used (2.5×10-5M). Then a gradual addition for CT-DNA amount till the
concentration raise from 0.00 to 1.58 × 0-4mol L-1 was done. The amounts used from CT-DNA, were
added also to each referenced cells include buffer solution to eliminate the absorption of free DNA.
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The intrinsic binding constant (Kb) of each compound towards DNA was extracted by using; [DNA] /
(єa − єf) = [DNA] / (єb − єf) + 1/Kb (єa − єf) equation [19]. Where; [DNA] is the molar concentration
of DNA, єa is the extinction coefficient observed for Aobs / [compound] at used DNA concentration.
Moreover, єf is the extinction coefficient for each free compound (4a-e), єb is the extinction
coefficient of the compound at full bonding towards DNA. Kb can be determined from the plots [DNA]
/ (єa − єf) vs. [DNA] through the ration; slope / intercept.
2.4. Analytical techniques
Elemental analysis of carbon, hydrogen, nitrogen, and sulfur of all compounds in concern, were
determined using Perkin-Elmer 2400 CHN Elemental Analyzer. Known referenced method [20], was
utilized to estimate the copper and chloride contents in synthesized complexes. The molar conductance
1
of tested complexes was performed by JENWAY model 4070 Conductance Bridge. HNMR and IR
(using KBr disc) spectra were conducted over Burker 400MHz and JASCO FT-IR-4100
spectrophotometers (400–4000 cm-1), respectively. Electronic absorption (UV/Vis) spectra (in DMSO)
and magnetic susceptibility were estimated at room temperature by applying UV2 Unicam UV/Vis
spectrophotometer and Johnson Matthey Magnetic Susceptibility Balance. Mass spectral analysis was
obtained at 70 ev by, AEIMS 30 mass spectrometer by heating rate 40°C/min, covering range of mass
50-1000. XRD patterns were obtained by using X-ray diffractometer (GNR, APD2000PRO, Italy) with
graphite mono-chromator over a range of 10˚ < 2θ < 90˚ at scanning rate by 0.03° min−1 under Cu/Kα1
radiation source. SEM images were extracted by implementing Joel JSM-6390 technique. TGA study
for complexes was executed by Shimadzu Thermal Analyzer by applying constant heating rate (10 oC
min-1) up to 20-900oC range, under N2. Conformational analysis and MOE docking were conducted by
Gaussian09 program and MOE module (version 2015), respectively. The biological activity test was
screened in specialized laboratory.
2.5. Conformational techniques
2.5.1. DFT/B3LYP method
All structural forms were adjusted by Gaussian 09 program [21], to build the best orientation for
functional groups, which confirms the bonding mode. The most fitted method was DFT using 3-21G
base set in ethanol, as a default solvent, in simulation with synthesis, which already done in it. The
basic files were acquired (log & chk) and visualized over Gauss-View program screen [22]. Basic
parameters were estimated based on EHOMO & ELOMO, and others were taken from files [23, 24].
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2.5.2. Molecular docking study
The crystal structures of 1miu, 4k9g and 5jm5 proteins in PDB format, were docked with most
synthesizes, for simulation. 1miu is a breast cancer susceptibility gene2 (1miu), 4k9g is a novel
inhibitor of macrophage migration, which shows efficacy in melanoma and colon cancer models, while
5jm5 is an AKR1C3 based Pro-drug TH3424, which has potent anti-tumor activity against liver cancer.
The introductory step, is the addition of hydrogen–atoms, after removing water molecules around the
helix. The charges and parameters were estimated by MMFF94x force field, after fixing the potential.
Implementing MOE module and after generating alpha-site sphere on site finder, the tested compounds
as MDB files, were docked on surface interior-grooves [25]. The scoring energy in MOE module, was
estimated using London dG scoring function and also upgraded by two unrelated-refinement by
triangle Matcher methods. The tested compounds were optimized, to minimize the energy content,
which led to best binding ability, after that the compounds oriented for analysis. Database browser was
used to compare the docking efficiency for tested co-crystals. From binding free energy score and
hydrogen bonds of co-crystals toward amino acids in protein-receptors, the rank of binding affinity,
was estimated. The hydrogen bonds were evaluated by measuring its length, which did not exceed than
3.5 Å. The type of bonded centers and the nature of bonding between crystal structures and protein-
receptors, were estimated, through mapping the receptor surface.
2.6. Usage of dyed-bandage in cancerous-wounds
2.6.1. Cell culture and viability
Human hepatic carcinoma (HepG-2), breast (MCF-7) colon (HCT116) and normal fibroblast cells
(HSF) were obtained from Viscera (Giza, Egypt). The cytotoxicity and anticancer activities of
prepared Cu(II) complexes (4a-e) were tested against three carcinoma cell lines in addition to one
normal cell line (HSF) using SulphoRhodamine-B (SRB) assay. Different cancer and normal cell lines
exposed to concentration range of, 0.01 to 100 µg/ml from complexes, then incubated (in 5% CO2
humidified incubator) at 37 °C for 72 h. Treated cells were fixed with TCA (10%) for 1h at 4°C.
Subsequently, to remove TCA, cells were washed with water many times, and then 0.4% SRB solution
was used to stain cells in dark place for 10 min. Stained cells washed with 1% glacial acetic acid.
Finally, to dissolve SRB-stained cells, Tris–HCl was used. After drying overnight, the color intensity
of remained cells was measured at 540nm by Elisa[26], in comparing to positive control (doxorubicin).
2.6.2. The impact of released complexes on cells
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In attempt to evaluate the impact of different Cu(II) complexes (4a-e) loaded on cotton, on colon(HCT-
116), liver(HepG-2) and breast (MCF-7) cancer cell lines as well as normal cell lines(HFS). All cells
were incubated with the same number of fibers (five fibers for each) isolated from the loaded cottons
for 72h in 5% CO2 humidified incubator. The cellular morphological-modifications were controlled
under inverted microscope every 24h, to get a correlation between, release amount of complex from
the cotton bandage, against the change of medium-pH. The data were analyzed using Sigma Plot
version 14.0.
3. Results and discussion
3.1. Analytical data
General characteristics of thiazole ligands and their Cu(II) complexes (4a-e) were displayed in Table 1.
The data presented confirm the formation of 2:1 (Cu:L) molar stoichiometry for all synthesized
complexes. The molecular formulae of all synthesizes were built based on elemental analysis and mass
spectral analysis for elected synthesizes, which matches directly to mathematical prediction. The
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complexes have non-hygroscopic nature, high m. p. up to 300 C and insoluble in traditional organic
solvents but excellently soluble in DMSO solvent. Two estimated values were 5.12 and 6.82 Ω-1 cm2
mol-1, for complexes Cu(II)-4a and Cu(II)-4d, respectively. This values support their non-electrolytic
nature, which agrees comfortably with covalent nature of chloride that analogues to its ionic one.
Whilst, Cu(II)-4e complex, displays molar-conductance value of 55.19 Ω-1 cm2 mol-1, which matches
conducting feature of one mobile anion [27].
3.2. Thiazole (4a-e) synthesis Mechanism
Thiazolyl-cyanoacetamides (thiazole-NHCOCH2CN) are known by their high reactivity with poly-
functional features. This due to possessing electrophilic and nucleophilic groups as, NH &CH2, which
arranged by, CH2 > NH. From this perspective, we synthesized new cyanoacetamido-thiazole derivatives
dyes (4a-e), with variable p-substituents, which changed between electron-donating to electron-
withdrawing groups, for discrimination. 2-Cyanoacetamido-thiazole (1), a starting material that was
prepared by heating 2-aminothiazole with cyanoacetic acid in the presence of acetic anhydride according
to referenced procedure [16]. The reactivity of active methylene-group in 2-cyanoacetamido-thiazole (1),
was tested towards variable-substituting diazonium salts. Diazocouplig reaction, which carried out
between cyanoacetamide (1) and diazonium salts derived from different aniline derivatives (in pyridine),
was ended by respective thiazolyl-hydrazonyl nitrile dyes, 4a-e (Scheme 1). The reaction was proceeded
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at most reactive site (methylene group) rather than another possible site (thiazole-C5). The analytical and
spectral data for compounds 4a-e, consistent with the proposed structures. IR spectra displayed
absorption bands at, 3460-3150, 2206-2220 and 1691-1652 cm-1 regions, which compatible with υN-H
or υO-H , υC≡N and υC=O groups, respectively. 1H NMR spectrum of 4c (as example) showed signals
confirming the atomic skeleton proposed. A singlet signal (3H) appeared at, δ= 3.78 ppm assigns for
methoxy group (-O-CH3). Doublet signals appeared at, δ= 7.00 and 7.80 ppm identified the aromatic
protons. While, the protons of thiazole-C5 and thiazole-C4 were resonated at 7.28 (doublet) and 7.55 ppm
(doublet), respectively. Moreover, a singlet signal appeared at, δ= 12.31 ppm assigns for NH or OH (in
enol tautomer).
3.3. The mode of bonding
In order to recognize the coordination sites in thiazole ligands, which chelate to Cu(II) ions, the IR
spectra of complexes. Cu(II)-thiazole complexes (4a-e) were contributed in deliberate comparison with
that of free ligands. Fundamental spectral bands for all couples (derivative and its complex) were
extracted and gathered in Table 2. With respect to thiazole derivatives, the bands appeared at; 3460-
3420, 3296-3150, 1655-1590, and 1691-1652 cm-1 ranges, assign for υ(OH) (in enol-form), υ(NH)s,
second υ(NH)s, nitrile function υ(C≡N), and υ(C=O), respectively. While, comparative study between
each free ligand and its coupled complex reveal such common information as;. i) Complete obscure for
υ(C=O) vibration, suggests its coordination with Cu(II) ions after de-protonation. ii)Also, vanishing
υNH (amide), is a further support for enolization during complexation process. iii) Non-obvious shift
of υ(C≡N) bands, denotes complete exclusion for CN group from coordination sphere. iv) ν(C-O)
vibration band which appeared at, 1160-1089 cm-1 range before complexation, after that, was appeared
with great shift in all complexes, due to coordination of enol-tautomer [28]. v) While, the vibration of
υ(C-S), which already presented at; 720-671 cm-1 rang, was appeared more-or less un-shifted in all
complexes. This a clear evidence for its ruling out from coordination, as theoretically expected. From
all previously reported, a mono-negative tetra-dentate mode of bonding contributing two central atoms
, was the only mode proposed for all complexes. Moreover, the appearance of new bands at; 691-675
and 860-812 cm-1 ranges, that assign to δw(H2O) and δr(H2O) of occluded solvent molecules,
respectively [29]. In addition to new vibrational bands at, 578-566 and 506-492 cm-1, which may
attribute to ν(M-O) and ν(M-N), respectively, were the final evidence for coordination feature.
3.4. Electronic spectral inspection
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Essential spectral bands for all investigated compounds were extracted from UV-Vis spectra to assert
on structural formula of interested complexes. Moreover, other spectral data as, oscillator strength, and
molar absorpitivity, were estimated for perfectly soluble compounds for 1x10-4 ML-1(in DMSO)
concentration (Table 3). Most of electronic transitions inside thiazole ligands, were obviously affected
by p-substituent's especially bands related to C=N group. π→π* transitions, which observed at 35,471-
37,120 and 28,710-35,184 cm-1, can be attributed to C=O and C=N bands, respectively. Furthermore,
n→ π* transitions were appeared within 22,910-30,178 cm-1 range. Such range, is highly contributing
with visible region, which considered the logical result for high conjugation inside ligands-structures
[30]. In common, blue shifts were observed for all intra-ligand transitions upon complex formation,
which seems to be an excellent evidence for coordination process. Also, the values of molar
absorpitivities computed for the first transition inside ligands, suffer reduction in the corresponding
complexes due to charge transfer transitions (L→M). Two structural geometries were suggested for
five inspected Cu(II) complexes; square-planer configurations for Cu(II)-4a, Cu(II)-4d, and Cu(II)-4e
complexes. While, an octahedral configuration for Cu(II)-4b, and Cu(II)-4c complexes. These variable
features originally depend on various recorded electronic transitions, which strongly conjugating with
p-substituent's effect. Spectra of [Cu2Cl3(PTAC)(H2O)]H2O, [Cu2Cl3(AcTAC)(H2O)]H2O, and
[Cu2Cl2(CTAC)(H2O)2]Cl.H2O complexes exhibited significant d-d transitions consequently at,
2
18,671;16,512, 18,326;16,521, and 18,056; 16,452 cm-1. These transitions assign to B1g→2A1g and
2B1g→2Eg electronic transitions, respectively [31]. These transitions confirmed the geometry of
square-planer arrangement proposed. Whilst, the spectra of [Cu2Cl3(TTAC)(H2O)4]2H2O, and
[Cu2Cl2(CTAC)(H2O)2]Cl.H2O complexes displayed two significant transitions at, 21,218;18,821 and
,
22,451;19,193 cm-1, respectively. These bands were assigned to 2B1g→2Eg and 2B1g→2B2g transitions,
which supporting distorted octahedral arrangements around cooper atoms [32]. Magnetic moment
values of the five complexes (ꢀeff =1.60-1.63 BM) were found less than the well-known values for one
unpaired electron [32]. These subnormal values, may be point to significant metal-metal interaction
inside binuclear complex. All optimized structural forms for Cu(II) complexes, were displayed in
figure 2.
3.5. Mass spectral analysis
To be satisfied with the chemical formulae proposed, the mass spectral analysis was performed for two
representative examples (Fig. 3), up to m/z = 1000. H-PTAC (4a) was the selected derivative for the
first mass analysis and its spectrum displayed, a molecular ion peak (M+) with exact mass at m/z=
271.05(100%)(calcd. 271.30). Also, the apparent split of molecular ion peak, which recorded at,
272.06(13.1%), 273.05(4.8%), 272.05(2.6%) and 273.06(1.1%), may point to M++1 and M++2.
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Through the influence of electron bombardment, the most fitted fragmentation pathway was exhibited
in Scheme 2. The other spectrum was executed for Cu(II)-4c complex, to serve as a common evidence
for all formulae proposed. The exact molecular ion peak was recorded at m/z = 639.25(2.4%)(calcd.
641.86), which attributing to M++2. This appearance may base on the isotopic nature of some integral
atoms as, chloride and copper-centers. Such base was supported by the appearance of two peaks at m/z
= 66.09 and 63.14, which attributing to copper isotopes [33].
Scheme. 2
3.6. XRD and SEM analysis
XRD patterns were performed within the range of 10˚ < 2θ < 90˚ (Fig. 4 &3S), to establish a broad
view about crystal lattice dynamics of solid samples. Also, the patterns clarity reflect the purity of
synthesized compounds from starting-reactants, based on referenced method [34]. All of synthesis
were appeared in crystalline nanoparticle, except Cu(II)-4b complex, which appeared in amorphous
morphology. Such crystalline feature, reflects the ideal packing of molecules inside the lattice, which
facilitate the calculation of lattice-parameters through known relations [35]. Using FWHM method, the
2θ, d spacing, relative intensity, particle sizes, crystal strain (ε) and dislocation density (δ), were
calculated and presented in Table 4. All particulate-sizes calculated, were become excellently in
nanometer range, with superiority of Cu(II)-complexes than corresponding-ligands. Also, Cu(II)-
complexes displayed reduced d-spacing values than the ligands, such reflects the impact of copper
metals on crystal-network packing. With respect to thiazole derivatives, the values of crystal strain and
dislocation density, reflect the uniformity and relatively perfect crystals of their particles. However and
regarding the complexes, the values of two parameters fall within the range of ideal crystals with few
imperfections which is acceptable and predictable with copper atoms. Moreover, the topography,
morphology and the microstructures of all synthesis, were tested and photographed (Fig. 4S) by
electron beam scanning for solid surfaces. Regarding thiazole derivatives, the particulate-features
appear by high extent of similarity, which logical with fixed moieties. While and regarding Cu(II)-
complexes, clear discrimination in all topographic features with that of free organic derivatives (4a-e),
assert on purity. The spherical particulate-shape, was commonly observed, which may serve perfectly
in broad spectrum of applications [36], as dyeing and antitumor-activity.
3.7. Thermogravimetric analysis
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All synthesized Cu(II)-thiazole complexes were tested in this analysis aiming the confirm their
suggested formulae(Fig. 5S). Also, discriminate the type of attachment for solvent-molecules towards
the coordination sphere. The most fitted postulation for thermal-decomposition pathway of all TG
curves, were aggregated in Table 5. All tested complexes were completely destroyed under the
influence of temperature up to ≈ 900 oC, mainly in four stages. All complexes were thermally-unstable
o
due to presence of crystal-water molecules, which easily released at low temperature (˂ 100 C).
Successive follower decomposition-stages, were attached with mass-loss, which apparently matching
with that proposed-theoretically. All decomposition-curves reached to steady state, which attributing to
the residual part, that include two copper atoms mainly as oxide.
3.8. CT-DNA binding
The degree of binding of most thiazole derivatives toward CT-DNA was studied spectrophotometry
using absorption titration method. Constant concentration was taken from each derivative and then,
regular increasing amounts from DNA were added at 25°C. Uv-Vis scanning curves (Fig. 6S) were
obtained over fresh solutions against reference solution for each concentration (blank). Each blank
solution, includes all additives except the organic amount, to remove the unwanted absorption from
excess DNA out of intended binding medium. The intrinsic binding constants (Kb) for tested
derivatives (all of them except H-PTAC) were estimated through an observable shift in definite charge
transfer(CT) band (see figure 4S, for pointed CT band). A significant hyperchromic effect as well as a
slight shift in a considered CT band(2-3nm), were observed attaching with increasing DNA amount.
This feature points to the stability of binding between the ligand and DNA helix. The spectral
appearance may refer to the electrostatic attraction or the occlusion of tested compound inside the
minor or major grooves. Furthermore, the hyper-chromic characteristic may be explained based on the
following; i) the elongated surface area for the tested compounds, which facilitate electrostatic
attraction over a whole DNA helix with various spots. ii) Occlusion of tested compounds inside the
grooves, led to re-organization for DNA helix. This need fractional release or damage for double helix
at the surfaces phosphate, leading to the formation of cavity appropriate for entering the compound.
Applying known spectral relationships [37], the binding constants (Kb, M−1) were calculated and
appeared as fellow; 3.92 x104(H-ATAC), 2.35 x105(H-TTAC), 4.92 x105(H-CTAC) and 9.64 x105 (H-
AcTAC). This behavior agrees well with Hammett's hypothesis[38]. The relation between Kb &
Hammett's constants (σR)(Fig. 5), introduces the influence of p-substituent on the degree of binding
(Kb).
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3.9. Conformational study
3.9.1. DFT/B3LYP method
The most fitted distribution for functional groups inside atomic-skeleton, was achieved for all new
synthesis (Fig. 1S & 2), by applying Gaussian09 program through density function method. Two
essential computed files were extracted as, log & chk. Such files were visualized over Gauss view
screen according to numbering scheme. Essential information were extracted from log files and
displayed in supplementary file (Table 1S). Whiles, other significant parameters (Table 6) were
computed based on frontier energy gaps (ꢁE = ELUMO -EHOMO). Also, HOMO & LUMO images for all
optimized structures were obtained from chk files and displayed (Figs. 6,7 & 7,8S). With respect to
thiazole-derivatives, HOMO& LUMO levels were appeared elongated over a whole molecule. Such
may attribute to distributed functional-groups, as well as high virtual conjugation. In addition to and
regarding Cu(II)-complexes, the two levels appeared in high similarity, and centered around central
atoms.
3.9.1.1. Estimated Physical parameters
Based on frontier energy gape (ꢁE) and using referenced relations [23,24], electronegativity (χ),
chemical potential (ꢀ), global hardness (η), global softness (S), global electrophilicity index (ω) and
absolute softness (ϭ) were estimated and presented (Table 6). With respect to Cu(II)-complexes and in
comparing with free derivatives, we concluded the following observations;. i) The values of
electrophilicity index (ω), are broadly elevated in complexes, than the corresponding derivative. Such
significant index, which considered the best indicator for the degree of toxicity and reactivity, clarified
the superiority of complexes. ii) The global hardness (η) and absolute softness (ϭ), which known as
opposite indexes, their values reflect the high softness of synthesized complexes, which favor in
biological activity [39,40]. The general enhancement recorded in physical characteristics of complexes,
may refer to the impact of central atom on exceeding the ꢁE values, which considered the indicator for
advancement [41]. Other physical parameters were extracted from log files and displayed in Table 1S.
Such parameters were as, bond lengths, selected atomic charges, Heat of formation (E), dipole moment
(D), oscillator strength (ʄ )and excitation energies(E). The contributing donor atoms (N &O) were
appeared have high negative charges, which considerably minimized after complexation. While, sulfur
atom appeared as positively charged, which pushed its exclusion from coordination. The bond-lengths
displayed, were selected to clarify lengthen of such bonds, after coordinating their donor side. The
noticeable reduction in the formation energy of complexes, reveal their high stability comparing to
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original derivatives. Also with respect to complexes, the general minimization of oscillator strength
values, reflect facilitated electronic-transition inside their molecular orbital building, as the influence
of metal atoms. Moreover, the lowering in dipole moment in complexes, may reflect extent of
covalence, especially with the presence of chloride atoms by covalent feature [42].
3.9.1.2. Electron density maps
The density maps of all optimized structures (Fig. 8 & 9S) were established after charge distribution
and building cubic surface. The electrophilic and nucelophilic attack-points, were resolute by their
electrostatic potential. The atomic-colors point to their degree of electron-density. With respect to blue
zone, which points to electron-poor region, which may face nucelophilic attack. While, the red zone,
points to electron-rich region, which may face electrophilic attack. Additionally, green color specified
to electrostatic potential region. The displayed maps supports by excellent way the ruling out of sulfur
atom from coordination due to its electrophilic nature with electron-lack.
3.9.2. Molecular Docking study
To give a good impact about the antitumor behavior of our new synthesis before actual study, we
interested to make such docking towards three tumor cell-lines as PDB for, 1miu, 4kg9 and 5jm5
proteins (breast, colon & liver), which will be screened actually. This aiming to investigate underlining
mechanism of binding mode and affinity between tested compound and amino-acids backbone. This
simulation process was performed for most new synthesis, from comparative point of view, by using
MOE module (v. 2015). With respect to docked thiazole derivatives (4b-e) towards three interested
proteins, we summarized the following notices as displayed in Table 7; i) the most contributing co-
crystallized ligand-centers were, N2, N12, S5, 5-ring and 6-ring towards various amino acids-
receptors. ii) The interaction types were covered all expected pathways as, H-π, π-π, H-donor and H-
acceptor. iii) The relative mean of standard deviation were found ≤ 4.0 . iv) The binding score values
(S ≈ -6.0 kcal/mol) reflect the degree of stability of docked complexes and can be arranged as such
descending order, 4d ˃ 4c ˃ 4b ˃ 4e. This arrangement was based on the revers relation among them
as, the degree of minimizing energy-score value, the high degree of stability for docking complexes
[43]. With respect to docked co-crystallized Cu(II)-complexes towards the same proteins, we
summarized the following notices as; i) the complexes were mainly bonded intensively than that of
free derivative and through most possible centers (Table 7). ii) Also, the interaction types were covered
all expected pathways as, H-π, π-π, H-donor and H-acceptor in addition to ionic binding. iii) The
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relative mean of standard-deviation were found more lengthen than that of corresponding derivatives,
which expected in coordinating compounds [44]. iv) The binding score values appeared comparable
with that of corresponding derivatives with superiority of Cu(II)-CTAC,4e complex. Regarding the
figures displayed (Figures 9& 10-12S), which clarify the binding centers of docked chelate with
various amino acids backbones, assert the degree of interaction in simulation process. With respect to
surfaces and maps (Figure 10) extracted for selected docking complexes, which assert on intensive
interacting centers in Cu(II)-complexes towards the electron density over amino acids in pathogen
proteins. Moreover, the docking simulation was also achieved for a reference drug (doxorubicin), for
ideal comparison. The interaction net result, may push the Cu(II)-CTAC,4e complex, as a strong
competitor in antitumor drug market. Finally, molecular docking study is considered the preliminary
step in drug industry, which may give a good impression about the true behavior, which may be
happened actually.
3.10. Successful usage of dyed- bandage for cancerous wounds
Utilizing SulphoRhodamine-B (SRB) assay, the cytotoxic impact of synthesized Cu(II)-thiazole
complexes on various cell lines as, MCF-7, HCT 116 and HepG2 in addition to normal cell line (HSF),
was evaluated. It was known that, the cellular-morphological changes, are commonly attached with
IC50 values. The tested complexes displayed less significant cytotoxicity and viability reduction in a
dose-response manner with very weak potential selectivity against tested different cell lines even the
normal cells (Table 8 & Figs. 11&13S). Interestingly, the highest anti-proliferative cytotoxic activity
of most complex, were detected against HCT116 colon cancer cell line with IC50 ranged from
33.6±2.82 to >1000±18. While, their impact against other cell lines, were ranged between moderate
and insignificant on all cell lines even normal cells. It is worthy to note that, the screening study was
conducted over pigmented cottons, to assess the extent of success, as bandage for cancerous-wounds.
The cellular-behavior against dyed fibers inside the media, was varied but interesting. MCF-7 went to
form colonies around fibers, the environment appears the same with HCT116 colon cancer, which was
very clear indicating that, the cells are more sensitive to fibers-dye. Whereas, the same cells displayed
another behavior against fibers not well dyed (Figs. 11& 13S). The most lighted point was, that the
normal cells did not present any reaction against the all fiber groups (dyed or not). Furthermore, the
use of fluorescent Acridine Orange (AO) and Ethidium Bromide (EtBr) mixed stain showed that cells
were attached to the fibers, which indicate that the fibers are not toxic at all. Effect in a biologic system
are not produced by a chemical agent unless the agent or its metabolite interact with appropriate
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receptors in the system in a sufficient time and concentration. Not only that, but also many medicines
are capable to be more active or may be lost their activity under the variation of body fluids conditions
like pH, or they may interact with other minerals or ions. In the light of these convictions, to keep
cancer cells under control, the present study suggested an innovative idea depends on the feature that
the microenvironment around the colony of cancer cells is generally acidic. Moreover, the acidity
condition may leads to breakdown the tight junction between cells, which may enhance the probability
of metastasis. The current study aimed to produce a trap bandage act associated with the increase of pH
in the microenvironment of the cancer cells colony to avoid the metastasis of released cells. Since it is
easy to monitor the pH change in cell culture media, the fibers were placed at the petri dishes at the
same time that the cells were cultured, to represent the model of metastatic cells. Therefore, this will
keep cancer cells in low activities. This release of dye (complex), may reflect its weak attachment with
the cotton fiber which already sensitive to pH value. Hence, the pigmented-bandage may contribute not
to eliminate cancer, due to weak toxicity recorded, but to reduce the activity of cancer cells through
regulating the pH.
4. Conclusion
New Cu(II)-thiazole complexes, were prepared and characterized using possible spectral and analytical
tools. The molecular and structural formulae were proposed. The structural forms were optimized by
using an advanced software, for conformation. The molecular docking was performed using MOE
module against selected three cancer proteins (1miu, 4k9g & 5jm5), to establish preliminary view about
the degree of inhibition for suggested drugs (synthesized compounds). A promising prediction for
treated complexes to be good tumor inhibitors, pushed us for antitumor application. Such application
was conducted by two ways, firstly by traditional way of application to calculate IC50. Secondly and
through unusual process, the screening was carried out on cotton fabrics after dyeing by these
complexes, to be tested as a bandage for cancerous wounds. The results were highly satisfactory in
achieving our goal.
References
[1] Ö. Şahin, Ü.Ö. Özdemir, N. Seferoğlu, Z.K. Genc, K. Kaya, B. Aydıner, S. Tekin, Z. Seferoğlu,
Journal of Photochemistry & Photobiology, B: Biology 178 (2018) 428–439
[2] G.M. Ziarani, R. Moradi, T. Ahmadi, N. Lashgari, RSC Adv., 2018, 8, 12069-12103
[3] M. Maeda, Laser Dyes, Academic, New York, 1994.
15

ACCEPTED MANUSCRIPT
[4] A. Gursoy, N. Karali, Turk. J. Chem. 27 (2003) 545–551
[5] B.Z. Kurt, I. Gazioglu, F. Sonmez, M. Kucukislamoglu, Bioorg. Chem. 59 (2015) 80–90.
[6] I.P. Ejidike, Molecules, 2018, 23, 1581-1998
[7] A.M.A.Alaghaz, H.A.Bayoumi, Y.A.Ammar, S.A.Aldhlmani, J. Mol. Struct. 2013, 1035, 383–399
[8] J.H. Al-Fahemi, A.M. Khedr, I.Althagafi, N.M. El-Metwaly, F.A. Saad, H.A. Katouah, Applied
Organometallic Chemistry, 32 (9), e4460 (2018).
[9] B. Iftikhar, K. Javed, M.S.U. Khan, Z. Akhter, B. Mirza, V. Mckee, J. Mol. Struct. 2018, 1155,
337–348.
[10] B. Naik, K.R. Desai, Indian J. Chem. 45 (B) (2006) 267–271.
[11] E. Wong, M. Christen Giandomenico, Chem. Rev. 99 (1999) 2451–2466.
[12] A.M. Florea, D. Büsselberg, review, Cancer 3 (2011) 1351–1371.
[13] Hanadi A. Katouah, Jabir H. Al-Fahemi , Marwa G. Elghalban, Fawaz A. Saad, Ismail A.
Althagafi, Nashwa M. El-Metwaly and Abdalla M. Khedr, Materials Science and Engineering: C,
, 96, 740–756 (2019).
I.Althagafi, N.M. El-Metwally, M.Elghalban, T.A. Farghaly, A.M. Khedr, Bioinorganic Chemistry
[14] and Applications, 2018, 1-15 (2018).
B.S. Creaven, B. Duff, D.A. Egan, Inorg. Chim. Acta 2010, 363, 4048–4058.
[15]
[16] H. M. Ibrahim, S. Makhseed, R. M. Abdel-Motaleb, A. A. Makhlouf, M. H. Elnagdi. Heterocycles,
2007, 71, 1951-1966.
[17] Zarkogianni Maria, Eleftheriadis Ioannis, Nikolaidis Nikolaos and Tsasaroni Eforia, Textile
Research Journal, 9,2014
[18] M. E. Reichmann, C. A. Rice, C. A. Thomos, P. Doty, J. Am. Chem. Soc., 1954, 76, 3047; J.
Marmur, J. Mol. Biol. 1961, 3, 208.
[19] A. Wolfe, G. H. Shimer, T. Meehan, Biochemistry, 1987, 26, 6392.
A. I. Vogel. Text Book Of quantitative Inorganic Analysis Longman, London 1986.
[20]
M. J. Frisch, et al., Gaussian 09, Revision D, Gaussian, Inc., Wallingford, CT, 2010.
[21]
R. Dennington, T. Keith , J. Millam .Gauss View, Version 4.1.2, SemichemInc, Shawnee Mission,
[22] KS, 2007.
U. El-Ayaan, N. M. El-Metwally, M. M.Youssef, S. A. A. El Bialy, Spectrochim. Acta, Part A,
16

ACCEPTED MANUSCRIPT
[23] 2007, 68, 1278.
[24] R. K. Ray, G. R. Kauffman, Inorg. Chem. Acta, 1990, 173, 207.
[25] Khlood H. Mashat , Bandar A. Babgi, Mostafa A. Hussien, Polyhedron 158 (2019) 164–172
[26] Gamil A. Al-Hazmi, Khlood S. Abou-Melha, Nashwa M. El-Metwaly and Kamel A. Saleh,
Bioinorganic Chemistry and Applications, 2018, Article ID 7176040, 22 pages
[27] W.J. Geary, Coord. Chem. Rev. 7 (1971) 81-122.
[28] A.M. Khedr, D.F. Draz, J. Coord. Chem., 63 (8), 1418–1429 (2010).
[29] K.S. Abu-Melhaa, N.M. El-Metwally, Spectrochim. Acta A 70 (2008) 277–283.
[30] A.A. Abou-Hussen, N.M. El-Metwaly, E.M. Saad, A.A. El-Asmy, J. Coord. Chem. 58 (2005)
1735-1749.
[31] A.M. Khedr, S. Jadon, V. Kumar, J. Coord. Chem., 64, 1351–1359 (2011).
[32] A. B. P. Lever, Inorganic Electronic Spectroscopy, Elsevier, Amsterdam 1986.
[33] M. S. Refat, N. M. El-Metwaly, Spectrochimica Acta Part A, 2011, 81, 215.
[34] B. D. Cullity, Elements of X –ray diffraction. Second ed., Addison –Wesley Inc., 1993.
[35] S. Velumani, X. Mathew, P.J. Sebastian, Sa.K. Narayandass, D. Mangalaraj, Sol. Cells, 2003, 76,
347.
[36] J. S. Ritch, T. Chivers, K. Ahmad, M. Afzaal, P. O. Brien, Inorg. Chem., 2010, 49, 1198; T.
Mokari, M. Zhang, P. Yang, J. Am. Chem. Soc. 2007, 129, 9864; J. J. Urban, D. V. Talapin, E. V.
Shevchenko, C. B. Murray, J. Am. Chem. Soc., 2006, 128, 3248.
[37] A.Z. El-Sonbati, A.F. Shoair, A.A. El-Bindary , M.A. Diab, A.S. Mohamed, J. Mol. Liq., 2015, 209,
635
[38] L. H. Abdel-Rahman, A. M. Abu-Dief, N. M. Ismail, M. Ismael, J. Inorg.Nano-Metal Chem. 2017,
47, 467
[39] R. C. Chikate, S. B. padhye, Polyhedron, 2005, 24, 1689.
[40] S.Sagdinc, B.Köksoy, K.F.andemirli, S.H. Bayari, J. Mol. Struct., 2009, 917, 63.
[41] I. Fleming, Frontier Orbital's and Organic Chemical Reactions, Wiley, London, 1976.
[42] S. K.Tripathi,R.Muttineni, S. K. Singh, J. Theor. Biol., 2013, 334, 87.
[43] Ahmed A. Fadda, Ehab Abdel-Latif,* Ahmed Fekri, and Amal R. Mostafa, J. Heterocyclic Chem.,
00, 00 (2019).
[44] Somaia S. Abd El-Karima, Manal M. Anwara, Yasmin M. Syama, Manal A. Naelb, Hanan F.
Alia, Mohamed A. Motaleb, Bioorganic Chemistry 81 (2018) 481–493
[45] C. Fosset, B. A. McGaw, and M. D. Reid, Journal of Inorganic Biochemistry, vol. 99, no. 5, pp.
1018–1022, 2005
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ACCEPTED MANUSCRIPT
[46] Gamil A. Al-Hazmi, Khlood S. Abou-Melha, Nashwa M. El-Metwaly and Kamel A. Saleh,
Bioinorganic Chemistry and Applications, Volume 2018, Article ID 7176040, 22 pages
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Table 1. Analytical characteristics for synthesized Cu(II)-thiazole complexes
Λm, Ohm-
Compounds
Color
Elemental analysis (%) Calcd (Found)
Cu
1cm2 mol-1
(Empirical formula)
C
H
N
S
Cl
------
---
5.12
---
1)(C12H9N5OS)(H-PTAC,4a)(271.30)
2)[Cu2Cl3(PTAC)(H2O)]H2O(539.77)
3)(C13H11N5OS)(H-TTAC,4b)(285.33)
4)[Cu2Cl3(TTAC)(H2O)4]2H2O(625.86)
5)(C13H11N5O2S)(H-ATAC,4c)(301.33)
6)[Cu2Cl3(ATAC)(H2O)4]2H2O(641.86)
7)(C14H11N5O2S )(H-AcTAC,4d)(313.34)
8)[Cu2Cl3(AcTAC)(H2O)]H2O(581.81)
9)(C12H8ClN5OS)(H-CTAC,4e)(305.75)
10)[Cu2Cl2(CTAC)(H2O)2]Cl.H2O(592.24)
Orange
53.13(53.13) 3.34(3.33) 25.81(25.80) 11.82(11.80) -----
23.55(23.56) 19.70(19.71)
Olive green 26.70(26.65) 2.24(2.24) 12.97(12.97) 5.94(5.92)
Golden
Brown
Golden
54.72(54.70) 3.88(3.85) 24.55(24.53) 11.24(11.22) ------
-----
---
24.95(24.94) 3.54(3.55) 11.19(11.18) 5.12(5.13)
20.31(20.31) 16.99(17.03)
---
51.82(51.80) 3.68(3.67) 23.24(23.24) 10.64(10.65) ------
------
---
Brownish yellow 24.33(24.31) 3.45(3.44) 10.91(10.90) 5.00(5.01)
19.81(19.81) 16.57(16.55)
---
Violet
53.67(53.66) 3.54(3.52) 22.35(22.33) 10.23(10.27) -----
------
6.82
---
Olive green 28.90(28.92) 2.43(2.44) 12.04(12.02) 5.51(5.49)
21.84(21.88) 18.28(18.30)
Red
47.14(47.15) 2.64(2.64) 22.91(22.91) 10.49(10.48) ------
------
55.19
Olive green 24.34(24.33) 2.38(2.37) 11.82(11.81) 5.41(5.41)
21.46(21.46) 23.94(23.95)
Table 2. Functional IR spectral bands (cm-1) for thiazole derivatives (4a-e) and their Cu(II) complexes
Compounds
δNHs
δr(H2O),
δw(H2O)
νOH,νNHs
νC=O νC=N
νC-S νC-O νCN
νM-O νM-N
3420,3215,3150 1690 1633, 1594 670 1093
3430,3174 ---- 1616,1605 677 1078
3457,3228,3197 1691 1636,1602 720 1160
B.C.at 3460, 3230 ---- 1620, 1594 719 1137
3431,3296,3187 1652 1623, 1590 696 1091
B.C. at 3452, 3170 ---- 1610, 1578 690 1082
3460, 3219, 3176 1688 1650,1610 671 1089
3465, 3190 --- 1641, 1593 681 1070
3428, 3240,3181 1689 1655,1601 678 1090
2217 1534, 1554
2219 1534
1)(C12H9N5OS)(H-PTAC,4a)
2)[Cu2Cl3(PTAC)(H2O)]H2O
3)(C13H11N5OS)(H-TTAC,4b)
4)[Cu2Cl3(TTAC)(H2O)4]2H2O
5)(C13H11N5O2S)(H-ATAC,4c)
6)[Cu2Cl3(ATAC)(H2O)4]2H2O
7)(C14H11N5O2S )(H-AcTAC,4d)
8)[Cu2Cl3(AcTAC)(H2O)]H2O
9)(C12H8ClN5OS)(H-CTAC,4e)
10)[Cu2Cl2(CTAC)(H2O)2]Cl.H2O
860, 691 570 492
2220 1585, 1561 -----
----
2225 1588
854, 676
572 505
----
2206 1580, 1526 -----
2210 1530
841, 682
566 490
----
2210 1590, 1553 ----
2221 1555
823, 675
571 506
----
2220 1580, 1543 ----
3445, 3192
----
1646, 1590 688 1081
2230 1560
812, 681
578 492
B. c. Broad centered
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Table 3. Electronic transition bands (cm-1) for 10-4M thiazole derivatives (4a-e) and their Cu(II) complexes
Compounds
ꢀ(eff) ε1
ʄ (105) d-d transition Intraligand and charge
bands (cm-1) transfer(cm-1)
υ2 ; υ1
Complex
Geometry
(B.M.)
-----
1.63
-----
1.63
-----
1.68
6120 450.4
------
18,671; 16,512 35,510; 32,831; 30.045; 25,421
------ 37,120; 35,184; 32,160; 30,178
21,218; 18,821 36,459; 33,843; 31,671; 29,542
------ 36,910; 35,083; 33, 120; 29,720
22,451 ;19,193 36,751; 34,821; 32,845; 29,651
35,471; 30,123; 28,71, 25,501
------
1)(H-PTAC,4a)
8000 588.8
4120 473.8
----- -----
3890 511.3
----- -----
Square-planer
------
2)[Cu2Cl3(PTAC)(H2O)]H2O
3)(H-TTAC,4b)
Octahedral
------
4)[Cu2Cl3(TTAC)(H2O)4]2H2O
5)(H-ATAC,4c)
Octahedral
6)[Cu2Cl3(ATAC)(H2O)4]2H2O
------ 11,912 9123.3
------
36,124; 33,842; 30,348; 27,180; 22,910 ------
7)(H-AcTAC,4d)
1.60
----
15,120 11580.3 18,326; 16,521 36,032; 33,671; 30,219; 26,821
Square-planer
8)[Cu2Cl3(AcTAC)(H2O)]H2O
9)(H-CTAC,4e)
5120 606.8
5520 630.5
------
36,032; 34,910; 31,561; 28, 900; 24,029 ------
1.62
18,056; 16,452 35,751; 34,610; 30,871; 27,810
Square-planer
10)[Cu2Cl2(CTAC)(H2O)2]Cl.H2O
ε
, Molar absorptivity. ʄ , Oscillator strengths, ʄ, calculated using the following expression: ʄ=4.6×10−9 εmax υ1/2, where; εmax is the molar absorptivity of the band
maximum and υ1/2 is the band width at half-height expressed in wave numbers: C. J. Ballahusen, Prog. Inorg. Chem. 2.251(1960).
Table 4. XRD data for nano-crystalline compounds
δ(Å-2)
Intensity
Size (Å) 2θ
d-spacing
(Å)
FWHM
Compounds
ε
2.8611 31.72 408
0.9412 25.94 238
1.8811 25.62 146
2.3541 26.20 420
0.6757 32.36 226
2.8591 31.76 226
0.8221 32.62 392
1.4130 26.36 408
0.9245 32.50 364
2.8186
3.4321
3.4742
3.3986
2.7644
2.8152
2.7429
3.3783
2.7527
0.1178 0.1222
0.4377 1.1288
0.2216 0.2826
0.1733 0.1804
0.4944 2.1902
0.1182 0.1223
0.4007 1.4796
0.28697 0.5009
0.3574 1.1700
0.5263
1.5789
0.7895
0.6316
2.2316
0.5266
1.8353
1.0526
1.6316
1)(H-PTAC,4a)
2)[Cu2Cl3(PTAC)(H2O)]H2O
3)(H-TTAC,4b)
4)(H-ATAC,4c)
5)[Cu2Cl3(ATAC)(H2O)4]2H2O
6)(H-AcTAC,4d)
7)[Cu2Cl3(AcTAC)(H2O)]H2O
8)(H-CTAC,4e)
9)[Cu2Cl2(CTAC)(H2O)2]Cl.H2O
2

ACCEPTED MANUSCRIPT
Table 5. Decomposition assumption for Cu(II)-thiazole complexes
Compounds
Steps
Temp.
(°C)
range Decomposed
Weight loss;
Calcd(Found %)
1st
25.97- 178.91 -2H2O+0.5Cl2
180.12- 310.72 -Cl2+HCN
311.2- 561.96 -C3H2N2S
562.01- 850.41 -C8H5N2
CuO+Cu
13.24(13.25)
18.14(18.11)
18.18(18.18)
23.93(23.96)
26.51(26.50)
[Cu2Cl3(PTAC)(H2O)]H2O
2ed
3rd
4th
residue
1st
33.97- 194.01 -2H2O+HCN
195.22- 366.20 -Cl2+4H2O
367.15- 589.66 -0.5Cl2+C4H2N2
590.17- 866.61 -C8H7N2
CuO+CuS
10.08(10.05)
22.84(22.82)
18.14(18.15)
20.96(21.01)
27.99(27.97)
[Cu2Cl3(TTAC)(H2O)4]2H2O
[Cu2Cl3(ATAC)(H2O)4]2H2O
2ed
3rd
4th
residue
1st
37.31- 105.21 -2H2O
106.81- 331.76 -1.5Cl2+4H2O+HCN 32.01(32.04)
332.11- 546.81 -C9H7N2
547.10- 854.90 -C3H2N2S
2[CuO]
5.61(5.62)
2ed
3rd
22.31(22.33)
15.29(15.27)
24.79(24.74)
4th
residue
1st
45.26- 185.05 -2H2O+Cl2
186.11- 490.32 -0.5Cl2+HCN
491.05- 774.91 -C8H9N4S
5C+2[CuO]
18.38(18.39)
10.74(10.72)
33.22(33.22)
37.67(37.67)
[Cu2Cl3(AcTAC)(H2O)]H2O
[Cu2Cl2(CTAC)(H2O)2]Cl.H2O
2ed
3rd
residue
1st
28.91-97.83
-H2O+0.5Cl2
9.03(9.00)
2ed
98.51- 237.76 -Cl2+2H2O+HCN
238.07- 480.60 -C4H2N2S
481.10- 789.61 -C4H4N2Cl
3C+Cu+CuO
22.62(22.61)
18.60(18.58)
19.51(19.57)
30.25(30.24)
3rd
4th
residue
Table 6. Computed parameters (ev) for all optimized structures in ethanol solvent using DFT/B3LYP method
EH
EL
EH - EL El-Eh
µ
η
S(eV-1) ω
ϭ
x
Compound
4a, H-PTAC -0.22953 -0.08618 -0.1434 0.14335 0.157855 -0.15786 0.071675 0.035838 0.173828 13.95186606
Cu(II)-PTAC -0.23424 -0.2009 -0.0333 0.03334 0.21757 -0.21757 0.01667 0.008335 1.419817 59.9880024
4b, H-TTAC -0.22288 -0.08464 -0.1382 0.13824 0.15376 -0.15376 0.06912 0.03456 0.171022 14.46759259
Cu(II)-TTAC -0.15692 -0.12078 -0.0361 0.03614 0.13885 -0.13885 0.01807 0.009035 0.533462 55.34034311
4c, H-ATAC -0.22463 -0.08629 -0.1383 0.13834 0.15546 -0.15546 0.06917 0.034585 0.174699 14.4571346
Cu(II)-ATAC -0.23763 -0.21906 -0.0186 0.01857 0.228345 -0.22835 0.009285 0.004643 2.807832 107.7005924
4d, H-AcTAC -0.23463 -0.09219 -0.1424 0.14244 0.16341 -0.16341 0.07122 0.03561 0.187467 14.04099972
Cu(II)-AcTAC -0.23254 -0.19723 -0.0353 0.03531 0.214885 -0.21489 0.017655 0.008828 1.307719 56.64117814
4e, H-CTAC -0.22929 -0.08898 -0.1403 0.14031 0.159135 -0.15914 0.070155 0.035078 0.180486 14.25415152
Cu(II)-CTAC -0.20404 -0.14269 -0.0614 0.06135 0.173365 -0.17337 0.030675 0.015338 0.489901 32.599837
3

ACCEPTED MANUSCRIPT
Table 7. The energy score and interaction data for most compounds against, breast, colon and liver cancers proteins
Compound
Protein Ligand
Receptor
Interaction Distance E( Kcal/ S(energy score)
mol)
4b, H-TTAC
1miu
4k9g
-5.3
-0.5
-0.6
0.0
N 13
N 12
N 6
5-ring
N 2
O
N
VAL 2967 (A)
PRO 1 (B)
H-donor
H-acceptor 3.33
H- π
π- π
H-acceptor 3.05
H- acceptor 3.58
π-H
π- π
H-donor
H- donor 2.70
ionic
2.96
-5.5643
-5.2706
6-ring PHE 113 (B)
6-ring TYR 36 (B)
N
N
4.19
3.45
5jm5
1miu
-0.9
-1.7
-0.8
0.0
LYS 270 (B)
TYR 24 (B)
-6.1201
-6.3179
N 12
5-ring
6-ring
N 15
O 25
O 46
O 49
O 49
N 14
N 15
O 49
O 25
6-ring
5-ring
N 2
5-ring
N 12
5-ring
CL 40
O 53
N 2
O 47
6-ring
O 50
N 15
6-ring
CA SER 217 (B)
6-ring TYR 24 (B)
OD1 ASP 41 (B)
OE2 GLU 40 (B)
OD1 ASP 41 (B)
OE1 GLU 40 (B)
OD1 ASN 109 (C)
CE LYS 77 (A)
OE1 GLU 237 (A)
OE1 GLU 237 (A)
OE1 GLU 237 (A)
CD PRO 239 (A)
CA GLN 2985 (A)
ND2 ASN 97 (C)
CB SER 60 (C)
4.10
3.96
3.28
Cu(II)-TTAC
-1.4
-5.6
-5.6
-3.7
2.84
3.12
2.74
3.84
3.05
ionic
4k9g
5jm5
-3.5
-0.5
-6.2
-4.8
-0.7
-0.5
H-donor
H-acceptor
H-donor
-5.3831
-5.4438
H- donor 2.94
ionic
π-H
π-H
H-acceptor 3.20
π-H
3.88
4.58
3.70
4c, H-ATAC
Cu(II)-ATAC
1miu
4k9g
-0.8
-0.5
-0.6
-1.0
-1.4
-0.5
-1.9
-1.5
-1.5
-6.3173
-6.7481
3.71
3.33
3.79
3.68
3.10
3.62
2.89
5jm5
1miu
N
LYS 270 (A)
H-acceptor
π-H
-6.372
CB SER 221 (A)
OE1 GLU 2641 (A) H-donor
OD1 ASP 2613 (A) H-donor
OE1 GLU 2479 (A) Ionic
OE1 GLU 2479 (A)
NE ARG 11 (A)
-5.6385
ionic
π- cation 4.21
H-donor
H-acceptor
π-H
4k9g
5jm5
-0.7
-15.0
-2.4
-0.7
-4.9503
-5.3137
2.80
3.25
4.63
O
GLU 237 (A)
OH TYR 196 (A)
CYS 7 (B)
OD2 ASP 41 (B)
N
4d, H-AcTAC
1miu
4k9g
4.11
4.36
3.20
3.09
3.67
-0.9
-1.0
-1.3
-0.8
-0.5
S
5
H-donor
-5.0058
-6.4142
6-ring
NH1 ARG 2926 (A) π -cation
S
5
OG SER 60 (C)
NE2 HIS 62 (A)
CB SER 60 (B)
ND2 ASN 280 (B)
NZ LYS 270 (B)
CD1 LEU 219 (B)
CB LYS 270 (B)
OE1 GLU 2824 (A)
5-ring HIS 2817 (A)
H-donor
H-acceptor
H- acceptor
H-acceptor
H- acceptor
π-H
π-H
Ionic
π- π
H-acceptor
H-donor
H-acceptor
H- acceptor
π-H
π-H
H- π
N 11
N 12
N 2
5jm5
3.42
3.12
3.92
3.78
-1.2
-1.7
-0.5
-0.6
-6.3778
N 12
5-ring
6-ring
O 45
6-ring
O 26
N 13
N 15
O 26
6-ring
6-ring
Cu(II)-AcTAC
1miu
2.95
3.77
2.94
3.09
3.56
2.97
3.14
3.19
-7.8
-0.0
-4.8
-7.5
-0.8
-2.2
-0.8
-0.5
-5.5061
4k9g
5jm5
N
O
PRO 1 (B)
ASP 156 (B)
-5.8605
-5.5401
CA LYS 136 (A)
NH1 ARG 96 (B)
CB ASN 134 (A)
CA ALA 157 (B)
5-ring HIS 2791 (A)
4e, H-CTAC
1miu
3.96
-0.7
C
15
-4.9259
4

ACCEPTED MANUSCRIPT
4k9g
5jm5
1miu
3.24
------
3.21
3.06
2.70
2.70
-6.4
------
-10.3
-2.3
-2.6
-6.9
N 12
------
N 13
N 16
O 40
O 40
N
------
OD2 ASP 41 (B)
OE2 GLU 40 (B)
OE2 GLU 40 (B)
OE2 GLU 40 (B)
PRO 1 (B)
H-acceptor
------
H-donor
H-donor
H-donor
ionic
-5.0639
-5.8681
-5.6783
Cu(II)-CTAC
4k9g
3.26
3.41
2.74
3.87
3.48
-0.5
-1.1
-20.9
-0.8
-2.0
C
4
O
ALA 114 (A)
H-donor
H-donor
H-donor
ionic
-4.4128
N 13
O 43
N 2
O 43
N 7
CL 36
O 43
O 43
OG SER 111 (A)
O
O
OXT ALA 114 (A)
OD1 ASN 307 (B)
O
ALA 114 (A)
ALA 114 (A)
Ionic
5jm5
1miu
3.37
3.25
2.75
2.75
-2.0
-0.5
-18.2
-6.4
H-donor
H-donor
H-donor
ionic
-5.0724
-6.5117
HIS 304 (B)
OD1 ASP 309 (B)
OD1 ASP 309 (B)
Doxorubicin
3.42
2.97
3.46
4.69
-0.5
-3.0
-1.1
-0.5
O 19
O 29
O 38
6-ring
------
CA GLY 3059 (A)
NH1 ARG 2893 (A) H- acceptor
NZ LYS 2891 (A)
N
------
OD2 ASP 109 (A)
NE ARG 66 (A)
H-acceptor
H- acceptor
π-H
------
H-donor
H- acceptor
HIS 3060 (A)
4k9g
5jm5
------
2.86
3.05
------
-1.2
-1.3
-5.2151
-5.7434
O 29
O 19
Table 8: The IC50 (µg/ml) of different complexes against different solid tumor cell lines (HCT116, MCF-7, HepG-2)
and HSF as a normal cell line.
Item
HCT-116
MCF-7
HepG-2
HSF
Cu(II)-4a
Cu(II)-4b
Cu(II)-4c
Cu(II)-4d
Cu(II)-4e
Doxorubicin
>100±0.43
>1000±0.18
46.15±2.48
63.11±1.96
33.6±2.82
>300±2.21
>300±0.97
>300±0.90
>600±1.85
>1000±1.67
>200±2.43
>300±1.47
>200±0.85
>200±1.33
>1000±1.84 >400±1.58
>300±1.84
>200±0.51
>500±1.52
>200±1.63
0.3979±0.052 0.6025± 0.02 0.3428± 0.10 0.831± 0.21
5

ACCEPTED MANUSCRIPT
Scheme. 1. Synthesis procedure of N-aryl-2-oxo-2-(thiazol-2-ylamino)-acetohydrazonoyl cyanide derivatives 4a-e
Scheme. 2. The fragmentation pattern of thiazole derivative (4a, H-PTAC)

ACCEPTED MANUSCRIPT
Fig.1 . 1H NMR spectra of two chosen thiazole derivatives
1

ACCEPTED MANUSCRIPT
Fig. 2. Optimized structures of Cu(II)-thiazole complexes
2

ACCEPTED MANUSCRIPT
Fig. 3. Mass spectra of two selected compounds
Fig. 4. XRD patterns for nano-crystalline 4a, H-PTAC derivative and its Cu(II) complex
3

ACCEPTED MANUSCRIPT
Fig. 5 . Hammett's relation between p- substituent (
derivatives.
σ R) values vs intrinsic binding constants (Kb) of triazole
4

ACCEPTED MANUSCRIPT
Fig.6 . Frontier HOMO images of enol- form of thiazole derivatives
5