Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
Research paper
Ahmed M. Mansour
PII: S0020-1693(16)30543-6
DOI: http://dx.doi.org/10.1016/j.ica.2016.09.031
Reference: ICA 17276
Please cite this article as: A.M. Mansour, Spectroscopic, DFT, magnetic and biological activity evaluation of Pd(II),
Pt(II) and Ru(III) complexes of Nitazoxanide, Inorganica Chimica Acta (2016), doi: http://dx.doi.org/10.1016/j.ica.
2016.09.031
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
Spectroscopic, DFT, magnetic and biological activity evaluation of Pd(II),
Pt(II) and Ru(III) complexes of Nitazoxanide
Ahmed M. Mansour*
* mansour@sci.cu.edu.eg; inorganic_am@yahoo.com
* Corresponding author
1
Abstract
[M(NTZ)Cl2] (M = Pd(II) (1), Pt(II) (2) and NTZ = Nitazoxanide) and [Ru(NTZ)Cl3(OH2)] (3) complexes
have been synthesized and characterized by elemental analysis, thermogravimetric analysis, IR, 1H NMR,
conductance and magnetic measurements. NTZ is coordinated to the metal ions as a bidentate ligand via
the N-atom of the thiazole ring and amide C=O. The susceptibility of Staphylococcus aureus and Escherichia
coli towards NTZ and its complexes has been evaluated. The calculated electronic structures and the
related electronic transitions have been obtained at TD-DFT/B3LYP/LANL2DZ level of theory. Upon the
chelation, the value of the natural charge of Ru(III) ion (+0.182e) is lower than those of Pd(II) and Pt(II)
ions (~+0.398e) in their complexes, which reflects the higher complexation ability of NTZ towards the
high oxidation state and low-d population metal ion. Complex 3 is existing in a low-spin octahedral
arrangement. The DC magnetic measurements in terms of Weiss constant revealed the presence of the
intermolecular interactions among the spin carriers. The low stability of 1-3 in DMSO and the rate of the
releasing of NTZ was the factor which reproduces the order of the toxicity (NTZ > 2 > 3 > 1) against the
tested microbes.
Key Words: Nitazoxanide; low-spin Ru(III) complexes; magnetic; NBO; TD-DFT; Antibacterial activity
* Corresponding author, Tel.: +2 2 01222211253; Fax: +20 2 35728843. E-mail: mansour@sci.cu.edu.eg; inorganic_am@ yahoo.com
2
Introduction
used in the treatment of different strains of bacteria [1, 2] and parasites such as Cryptosporidium parvum
[3], and Giardia lamblia [4]. Its mechanism of action involves the inhibition of ferredoxin oxidoreductase
enzyme-dependent electron transfer reaction that is essential for the anaerobic energy metabolism [5]. In
2009, the anti-Influenza properties of NTZ, against oseltamivir-resistant H1NI, and influenza B virus, was
introduced through the blockade of the maturation of influenza [6, 7]. Several analytical methods were
used for the determination of NTZ and its metabolic products [8-16]. Tizoxanide (TZ), one of the major
degradation products of NTZ, was identified and characterized using different spectral tools [8]. TZ and its
glucuronide form were rapidly and specifically determined in mouse plasma by LC-MS/MS [9, 10], UPLC-
MS/MS [11] and LC [12] methods. HPLC was used to determine the concentration of NTZ in presence of its
degradation products, which were produced by either the oxidative, acidic or basic reaction [13]. Spectral
tools were used for the quantitative assay of NTZ [14, 15]. For example, a simple extractive method based
upon the formation of colored ion associate complexes with N-bromosuccinimide and indigo carmine, was
used for the assay of NTZ [14]. The determination of NTZ was electrochemically achieved by square wave
voltammetry using a fabricated electrode based on the synergistic effect of graphene and polyaniline
The structural and thermal properties of NTZ were determined and studied by Sperandeo et al. [17].
Geometry optimization and the normal coordinate analysis of NTZ were also reported [18]. Several co-
crystals of NTZ with succinic acid and glutaric acid were isolated and characterized by single-crystal X-ray
diffraction analysis, infrared and thermal methods [19]. Only the coordination behavior of NTZ towards
Mn(II), Fe(III), Cu(II) and Zn(II) [20] as well as Ag(I) [21] were touched. Octahedral geometries were
suggested for the di- and trivalent metal ions. NTZ was assumed to interact as a bidentate drug via NH and
In view of the appearance of methicillin resistant strains of Staphylococcus aureus [22], growing
bacterial resistance opposing effects towards some antibiotic drugs [23], and the low bioactivity, the
3
search for the new effective antimicrobial agents is of vital importance. However, the finding of new drugs
having new chemical structures is very expensive and time consuming. Thus, the molecular structure
modification is better and desirable especially when the physicochemical properties are changed with the
improvement of the drug-acceptor interactions. One of the structural modification method is the complex
formation. For example, coordination of the well-known antibacterial drug, nifuroxazide, to Pd(II) ion led
to an improvement in the antibacterial activity [24], where the LC50 value decreases from 0.072 µmol/mL
to 0.029 µmol/mL upon the complexation. Here, new Pd(II), Pt(II) and Ru(III) complexes of NTZ (Scheme
1) have been synthesized and characterized. The experimental studies have been accompanied by density
has been investigated in the temperature range of 77-298 K. Finally, the susceptibility of Staphylococcus
aureus and Escherichia coli towards NTZ and its complexes has been evaluated.
Experimental
Instruments
FT IR spectra were recorded as potassium bromide pellets using a Jasco FTIR 460 plus in the range
of 4000 to 200 cm-1. 1H NMR spectra were run at 300 MHz in DMSO-d6 using a Varian-Oxford Mercury VX-
300 NMR. 1H-NMR spectra were recorded also in acetone-d6 and methanol-d4 by Bruker Avance 400
nitrogen atmosphere (20 mL min-1) in a platinum crucible with a heating rate of 10 ºC min-1 using a
Shimadzu DTG-60H simultaneous DTG/TG apparatus. A digital Jenway 4310 conductivity with a cell
constant of 1.02 was used for the molar conductance study. Elemental microanalysis was performed using
Elementer Vario EL III. Electronic spectra were scanned on OPTIZEN POP automate spectrophotometer in
temperature range at a magnetic field H = 2 kG. Diamagnetic corrections were made by Pascal's constant
Synthesis
4
The solid complexes 1-3 were synthesized by adding NTZ (307 mg, 1 mmol) in acetone (15 mL) to the
aqueous solution (5 mL) of Na2PdCl4 (294 mg) (1), and K2PtCl4 (415 mg) (2) or the acetone solution (5
mL) of RuCl3·3H2O (261mg) (3). The reaction mixtures were heated to reflux (80 ºC, 2 h (1), 4h (2) and 4h
(3)), whereupon orange, yellow and dark brown complexes were precipitated. Lower conductance values
in DMF were reported for 1 and 3 compared with the reported values [25] for 1:1 (65-90 Ω-1 cm2 mol-1)
and 1:2 (130-170 Ω-1 cm2mol-1) electrolytes in DMF, indicating their non-ionic nature. The unexpected
higher conductance value of the Pt(II) complex 2 may be attributed to its instability and decomposition
upon the dissolution in DMF. This has been confirmed by the 1H NMR measurement (discussed later). The
conductance values were also recorded in methanol and compared with that of the free solvent (54.7 Ω-1
cm2 mol-1). Lower differences were reported as complexes 1-3 have the conductance values of 63.0, 57.7
− Complex 1: Color: Orange. Elemental analysis (%): calcd. C12H9Cl2N3O5PdS: C 29.74, H 1.87, N 8.67,
found C 29.66, H 2.13, N 8.43. IR (FT IR, cm-1): 3431 (br, N-H), 1763 (vs, C=O), 1646 (vs, C=O), 1550
(m, C=N), 1490, 1436, 1355, 1290, 1095, 918, 736, 594. 1H NMR (acetone-d6, 400 MHz, δ, ppm):
10.71 (s, 1H, NH), 8.11 (s, 1H, thiazole), 7.81 (d, 1H, CHAr), 7.63 (t, 1H, CHAr), 7.32 (t, 1H, CHAr), 6.96
(d, 1H, CHAr), 2.52 (s, 3H, CH3). 1H NMR (methanol-d4, 400 MHz, δ, ppm): 8.35 (s, 1H, thiazole), 7.71
(d, 1H, CHAr), 7.57 (t, 1H, CHAr), 7.35 (t, 1H, CHAr), 7.18 (d, 1H, CHAr), 2.20 (s, 3H, CH3). UV-vis. (DMF,
10-5, 10-4, nm): 295, 400, 435, 495. Molar Cond. (10-3 M, DMF, Ω-1cm2 mol-1): 7.15.
− Complex 2: Color: Yellow. Elemental analysis (%): calcd. C12H9Cl2N3O5PtS: C 25.14, H 1.58, N 7.33,
found C 24.93, H 1.49, N 7.17. IR (FT IR, cm-1): 3431 (br, N-H), 1765 (vs, C=O), 1648 (vs, C=O), 1551
(m, C=N), 1489, 1437, 1355, 1315, 1219, 1194, 1096, 1012, 919, 796. 1H NMR (acetone-d6, 400 MHz,
δ, ppm): 12.55 (s, 1H, NH), 9.05 (s, 1H, thiazole), 8.52 (d, 1H, CHAr), 8.26 (t, 1H, CHAr), 8.01 (t, 1H,
CHAr), 7.89 (d, 1H, CHAr), 2.60 (s, 3H, CH3). 1H NMR (methanol-d4, 400 MHz, δ, ppm): 8.33 (s, 1H,
thiazole), 7.70 (d, 1H, CHAr), 7.55 (t, 1H, CHAr), 7.32 (t, 1H, CHAr), 7.16 (d, 1H, CHAr), 2.18 (s, 3H, CH3).
UV-vis. (DMF, 10-5, 10-4, nm): 365, 435, 520. Molar Cond. (10-3 M, DMF, Ω-1cm2 mol-1): 127.5.
− Complex 3 Color: Dark brown. Elemental analysis (%): calcd. C12H11Cl3N3O6RuS: C 27.05, H 2.08, N
7.89, found C 26.90, H 1.86, N 7.79. IR (FT IR, cm-1): 3433 (br, N-H), 1763 (w, C=O), 1644 (vs, C=O),
1553 (m, C=N), 1458, 1355, 1317, 1255, 1206, 1165, 1117, 1069, 874, 818, 656. UV-vis. (DMF, 10-5,
10-4, nm): 260, 345, 385, 430, 510. Molar Cond. (10-3 M, DMF, Ω-1cm2 mol-1): 18.0. μeff (μB, 298 K):
1.06.
5
DFT calculations
Gas phase geometry optimization of complexes 1-3 were carried out by density functional theory
(DFT), Becke 3-parameter (exchange) Lee-Yang-Parr (B3LYP) functional and the effective core potential
(ECP) of Hady and Wadt, LANL2DZ basis set [26] using Gaussian03 package [27]. The complexes have
been characterized as local minima via the harmonic frequency analysis and the IR spectra were analyzed
for the absence of imaginary modes. Natural bond orbital (NBO) analysis and frontier molecular orbitals
were performed at the same level of theory. The electronic transitions were calculated with TD-DFT/
B3LYP/LANL2DZ method in DMSO using the default polarizable continuum model (PCM) [28].
Antibacterial activity
The antibacterial activities were determined by a modified Kirby-Bauer disc diffusion method as
previously described [29] under the standard conditions using Mueller-Hinton agar medium (tested for
composition and pH), [30]. The antimicrobial activities were carried out using culture of Staphylococcus
The solid Pd(II) (1) and Pt(II) (2) complexes of the type [M(NTZ)Cl2] as well as [Ru(NTZ)Cl3(OH2)]
(3) have been synthesized from the reaction of Na2PdCl4, K2PtCl4 and RuCl3·3H2O with one equivalent of
NTZ (Scheme 1). Among the various IR active bands of NTZ spectrum, the comparison of the ester -COO,
amide C=O and NH as well as the thiazole C=N bands of the free NTZ with those observed in the complexes
were found to be useful in determining the active coordination sites in the studied drug. The IR spectrum
of NTZ (Fig. S1†) shows four characteristic stretching bands at 3357, 1774, 1661 and 1604 cm-1 allocated
for ν(NH), ν(C=O)ester, ν(C=O)amide and ν(C=N)thiazole. Upon the chelation, the ν(NH) mode is shifted to higher
wave numbers, which eliminates the involvement of the -NH- functional in the coordination sphere of the
metal ions. Alternatively, the ν(C=O)amide and ν(C=N)thiazole modes are moved to lower wave numbers that
may be taken as an evidence for the bidentate nature of NTZ. The ν(C=O)amide mode is observed at 1646,
1648 and 1644 cm-1 in 1-3, respectively, while the ν(C=N)thiazole is overlapped with the ν(C=C) vibrations
6
giving a new combination band in the range of 1553-1550 cm-1. The ν(C=O)ester vibration is observed in the
region of 1765-1763 cm-1, which suggests the participation of -COO- group in the intra- or intermolecular
The 1H NMR spectrum of NTZ (Fig. S2†) is characterized by three singlet signals at δ 13.58, 8.66 and
2.25 ppm corresponding to -NH, thiazole-CH and -CH3 as well as two doublets and two triplets at 7.86,
7.31, 7.71 and 7.47 ppm accounted for the phenyl ring. The synthesized diamagnetic complexes are
unstable in DMSO and they are decomposed to give the same positions and number of signals as those of
the free ligand. The 1H NMR spectrum of 2 is typical as that of the free ligand, while the spectrum of 1 is an
intermediate case between the free NTZ and the coordinated one. However, it is worthy to indicate that
complexes 1-2 show a signal corresponding to the amide-NH group, which excludes the possibility of its
enolization as a result of the complex formation. The 1H NMR spectra were recorded also in acetone-d6
and methanol-d4 (Fig. S3†). Both complexes are slightly soluble in these solvents. The downfield shift of
the thiazole aromatic proton and presence of a signal corresponding to the amide NH (in acetone) suggest
that NTZ interacts with thiazole ring without enolization of the NH group.
The absorption spectrum of free NTZ (Fig. S4†) in DMF exhibits three bands at 260, 345 and 435 nm
due to the low energy π-π* transition within the phenyl and thiazole rings [31], π-π* interaction of the C=O
groups and charge transfer band as a results of the strong withdrawing effect of the nitro group [24].
Complex 1 displays four transitions at 295, 400, 435 and 495 nm. The red shift of the band at 260 nm (in
NTZ) to 295 nm may be taken as an indication for the participation of thiazole-N in the complex formation.
The new band at 400 nm is allocated for MLCT transition. The lowest energy at 495 nm (logεmax = 3.07) is
ଵ
assigned to Aଵ → ଵBଵ (ν2) [24, 31] in the square-planar geometry. The absorption spectrum of 2 is
characterized by two broad bands at 365 and 435 nm due to the internal ligand transition. The electronic
spectrum of 3 shows five transitions at 260, 345, 385, 430 and 510 nm. The low-spin Ru(III) octahedral
ହ
complexes (S = ½) have the ground state symbol, 2T2g, which arises from ݐଶ electronic arrangement. Eight
transitions were expected as a result of transfer of one electron to the ݁ orbitals, and two transitions
7
were predicted as a result of the formation of the quartet state [32]. The first two excited doublet states
are 2A2g and 2T1g, respectively. The band at 385 nm may be assigned to MLCT [33], while this at 510 nm is
Magnetic properties
The temperature-dependent molar susceptibility measurement for the powder Ru(III) 3 sample
has been collected by DC-susceptibility experiment in the temperature range of 77-300 K using a field of 4
kG. The plots of the effective magnetic moment (µeff) and χm-1 versus T are represented in Fig. 2. It can see
that the µeff value of complex 3 decreases linearly with increasing the temperature from 1.35 µB at 77 K to
attain the minimum value (1.06 µB) at the room temperature that is in agreement with the paramagnetic
nature [34]. On the other hand, the χm-1 versus T plot is strictly linear and obeys the Curie-Weiss law [χm-1
= (T-θ)/C] with a Weiss constant θ of 28.19 K, and the Curie constant (C = N2g2β2S(S+1)/(3k)) is 0.16 emu
K/mol. The positive value of θ indicates the presence of intermolecular interactions among the spin
carriers and this was reflected in the value of µeff at the lower temperature. The µeff value of 3 [32, 35-37]
at the room temperature is lower than the expected value for the low-spin Ru(III) octahedral complexes
ହ
with only one unpaired electron (ݐଶ ). This may be explained in terms of the low symmetry of the ligand
field around the metal ion [38] and the presence of three chlorine atom in the coordination sphere, which
may decrease the electron density around the metal center and hence the magnetic moment such as
[Ru(SM)Cl2(OH2)2]·2H2O (SM: sulfamethazine) , 1.20 µB (296 K) [32], K2OsCl6 (1.44 µB, 300 K) [39] and
Thermogravimetric analysis
The TG curve of 1 displays two degradation events at 244 and 442 ºC in the range of 25-600 ºC
corresponding to the pyrolysis of one NTZ molecule with overall mass amounts to 63.44% (calcd. 63.59%)
giving palladium chloride salt as a final residue at 600 ºC (Fig. S5†). The thermogram of 2 shows no
inflection below 144 ºC, which reveal the absence of hydrated water molecules or solvent molecules. Two
decomposition steps maximized at 246 and 622 ºC. The 1st stage is assigned to loss of ester and nitro
8
groups as well as 2Cl with a mass loss of 30.70% (calcd. 30.71%). The 2nd stage brings the total mass loss
up to 65.89%. Such value is close to the calculated value (65.97%) expected for the formation of platinum
metal as a final residue at 900 ºC (Fig. S4†). The TG of 3 exhibits two steps at 260 and 610 ºC. The 1st stage
is accompanied by a mass loss amounts to 31.82%. This value finds a parallelism with the calculated value
(32.01%) responsible for the elimination of one coordinated water molecule, 3Cl and nitro group. The
second event is assigned to the pyrolysis of the rest of the organic part with overall mass loss amounts to
The optimized structures of [M(NTZ)Cl2] (M = Pd (1) and Pt(2)) as well [Ru(NTZ)Cl3(OH2)] (3) were
obtained by DFT/B3LYP/LANL2DZ method. The geometries of 1-3 and their atomic numbering are shown
in Fig. 2. The ground state geometries were characterized as local minima through the calculation and
analysis of the harmonic frequencies. The square-planar geometry of 1 and 2 is formed from a bidentate
NTZ drug [Pd-O(15) = 2.106 Å, Pd-N(25) = 2.107 Å (1) and Pt-O(15) = 2.076 Å, Pt-N(25) = 2.064 Å (2)]
and two chlorine atoms [Pd-Cl(32) = 2.355 Å, Pd-Cl(33) = 2.351 Å (1) and Pt-Cl(31) = 2.379 Å, Pt-Cl(32) =
2.381 Å (2)]. The complexes are coplanar, but the ester ring is bent away with a dihedral angle of 39.0 º to
relive the steric hindrance. The two Pd-Cl bonds are equal, but they are shorter than those of the Pt-Cl
bonds by about 0.03 Å. Similar, the Pt-O(15) and Pt-N(25) bonds are shorter than the palladium bonds,
which could be explained in terms of the metal size. The O(15)-Pt-N(25) bond (89.4 º) is slightly longer
than that observed in case of complex 1 (Table S1†), which may be ascribed to the dissimilarity in the
According to the natural bond orbital (NBO) analysis [41], the electronic configuration of Pd in 1 is
the formal charge of +2 as a result of the electron density donation form the four coordination centers.
The occupancies of Pd-4d sub-shell are ݀௫௬ 1.080݀௫௭ 1.954 ݀௬௭ 1.971݀௫ మି௬మ 1.951݀௭ మ 1.949. The σ(Pd-Cl(32)) bond is
formed from sp0.34d1.25 hybrid on Pd atom (38.62% s, 12.96% p and 48.43% d) and sp10 (90.91% p) on the
Cl(32) with a polarization of 73.75% towards the Cl(32) atom. The Pd-Cl(33) has the same hybridization
9
orbitals and polarization value as the Pd-Cl(32). For complex 2, the electronic arrangement of Pt is
[Xe]6s0.545d8.706p0.376d0.01, its charge is +0.382e and the occupancies of the d-orbitals are ݀௫௬ 1.034݀௫௭ 1.899
݀௬௭ 1.952݀௫ మି௬మ 1.925݀௭ మ 1.894. The (Pt-Cl(31)) bond is created from sp0.28d1.28 hybrid on Pt atom (38.93% s,
11.08% p and 49.99% d) and sp1.75 (87.74% p) on the Cl(31), and is polarized towards the Cl(31) atom
with about 71.36% of the electron density on the chlorine atom. Therefore, the Pd-Cl bonds are more
polarized than the Pt-Cl bonds. According to the second order interaction energy (E2) [26], the larger the
E2 value, the more intensive is the interaction between the metal and coordination sites. For 1, the E2
values are 7.86, 10.65, 7.89 and 7.75 cal/mol for LP(2)O(15)→σ*(Pd-Cl(32)), LP(1)N(25)→σ*(Pd-Cl(33)),
characterized by four interaction values, 14.38, 18.57, 3.28 and 3.02 cal/mol due to LP(2)O(15)→σ*(Pt-
comparison, it can see that the coordination sites (O(15) and N(25)) of NTZ prefer to coordinate to Pt(II)
The structure of 3 (Fig. 2c) could be described as a distorted octahedral geometry as can be seen
from the slightly spread in the optimized bond lengths (2.109-2.408 Å) and bond angles (85.9-97.6 º). NTZ
drug coordinates to Ru(III) ion as a bidentate ligand via thiazole-N [Ru-N(25) = 2.116 Å] and amide C=O
[Ru-O(15) = 2.109 Å]. Three chlorine ligands occupy fac-disposition of Ru(III) coordination sphere [Ru-
Cl(31) = 2.380 Å, Ru-Cl(33) = 2.408 Å, and Ru-Cl(34) = 2.435 Å]. One axial water atom [Ru-O(35) = 2.177
Å] competes the octahedral geometry. The natural charge of Ru is +0.182e and its electronic arrangement
is [Kr]5s0.284d6.865p0.645d0.046p0.01. The Ru-Cl(31) bond is formed from sp0.61d2.73 hybrid on the Ru atom
(23.03% s, 14.06% p and 62.90% d) and sp5.36 (84.27% p) with a polarization of 70.33% towards the Ru
atom. The other Ru-Cl bonds [Ru-Cl(33) and Ru-Cl(34)] have nearly the same hybrid orbitals, sp0.67d2.40 on
Ru (24.56% s, 16.56% p and 58.88% d) and sp4.70 on Cl (82.47% p). The M-L interaction energies values
(E2) are 8.47, 10.32, 3.73, 2.65, 2.98 and 9.05 cal/mol for LP(2)O(15)→σ*(Ru-Cl(34)), LP(1)N(25)→σ*(Ru-
To get an insight into the spectral properties and electronic structure of the complexes, TD-DFT
calculations were carried out by B3LYP/LANL2DZ method in DMSO using PCM model. The nature of the
electronic transitions has been assigned via the calculation of the lowest singlet spin-allowed excitation
states. The TD-DFT spectrum (Fig. S4†) of 1 shows five transitions in the UV-region at 228, 264, 298, 329
and 359 nm as well as two bands in the visible range at 514 and 629 nm corresponding to H-6→L+1, H-
6→L+2, H-1→L, H-2→L, H-3→L+1, H-5→ L+1 and H-4→L+1 (H: HOMO and L: LUMO), respectively. The
LUMO’s orbitals (LUMO+1 and LUMO+2) have contribution from the d-orbitals. The LUMO+1 orbital is
mainly formed from Pd ݀௭ మ , while LUMO+2 is ݀௫ మି௬మ in nature (Table S2†). Hence, the transitions at 359,
514 and 629 nm are LMCT, while those at 228 and 264 nm could be assigned to Br→Pd(II) interactions.
The other bands at 329 and 359 nm are π-π* within the organic part. Six main transitions are observed in
the stimulated spectrum (Fig. S5†) of complex 2 at 282, 335, 366, 441, 562 and 575 nm allocated for H-
6→L+1, H-7→L, H-6→L, H-1→L+2, H-1→L and H→L in that order. The descriptions of the frontier MO’s
and the relocation of the electron density of 2 are given in Fig. 3. The spectrum (Fig. S6†) of 3 displays two
main transitions at 591 and 421 nm corresponding to H-1(β)→ L+1(β) and H-2(β)→L+1(β) with oscillator
strengths of 0.0003 and 0.04. The lowest transition energy has a ground state composed of Ru-d character
, whereas the excitation state is of ݀௫ మି௬మ nature forming a d-d transition (Table S3†). The highest energy
band at 421 nm is LMCT band coming from the interaction of the thiazole ring with Ru(III) ion.
Antibacterial activity
The antimicrobial activities of NTZ and its complexes have been investigated against Staphylococcus
aureus (G+) bacterium and Escherichia coli (G-) microbe using a modified Kirby-Bauer disc diffusion method
[29, 42], and compared to tetracycline used as a standard. Preliminary screening was carried out at 20
mg/mL. The complexes exhibited the same activity against the microbes, regardless of its type of wall, and
the order of activity was found to be NTZ > 2 > 3 > 1 (Fig. 4). In absolute value, NTZ has the lowest MIC50
value of 26 µg/mL with respect to its complexes 1-3. This may be assigned to its low lipophilicity, where
the membrane penetration through the cell is decreased and hence it cannot block or inhibit the growth of
11
the microbes. The order of the complexes is related to the stability constant that is in agreement with the
previously discussed NMR data, where the truly toxic species of the drug is its free form.
Conclusion
The reaction of the antibacterial drug nitazoxanide with Pd(II), Pt(II) and Ru(III) ions afforded
complexes of the type of [M(NTZ)Cl2] (M = Pd(II) and Pt(II)) and [Ru(NTZ)Cl3(OH2)]. The solid state FT IR
suggests a bidentate nature of nitazoxanide. Geometry optimization, natural bond orbital analysis and TD-
DFT calculated electronic spectra have been obtained by DFT/B3LYP/LANL2DZ method. The Ru ion has
the lowest natural charge value with respect to Pd(II) and Pt(II), which reflects the preference of the drug
towards the high oxidation state and low-d population metal ion. The positive value of Weiss constant and
the low effective magnetic moment value indicates the presence of intermolecular interactions among the
spin carriers. The decomposition of the platinum complex upon the dissolution in DMSO and its high
toxicity against the tested microbes compared with the other complexes is the key that the truly toxic
References:
[1] L. Dubreuil, I. Houcke, Y. Mouton, J.F. Rossignol, Antimicrob. Agents Chemother. 40 (1996) 2266.
[2] J. Freeman, S.D. Baines, S.L. Todhunter, G.S. Huscroft, M.H. Wilcox, J. Antimicrob. Chemother. 66 (2011)
1407.
[3] A. Baishanbo, G. Gargala, C. Duclos, A. François, J.F. Rossignol, J.J. Ballet, L. Favennec, J. Antimicrob.
Chemother. 57 (2006) 353.
[4] B. Amadi, M. Mwiya, J. Musuku, A. Watuka, S. Sianongo, A. Ayoub, P. Kelly, Lancet 360 (2002) 1375.
[5] P.S. Hoffman, G. Sisson, M.A. Croxen, K. Welch, W.D. Harman, N. Cremades, M.G. Morash, Antimicrob.
Agents Chemother. 51 (2007) 868.
[6] J.F. Rossignol, S.F. La, L. Chiappa, J. Biol. Chem. 284 (2009) 29798.
[7] J.F. Rossignol, Antivirial Res. 110 (2014) 94.
[8] M.D. Malesuik, H.M.L. Goncalves, C.V. Garcia, M.R. Trein, N.B. Nardi, E.E.S. Schapoval, M. Steppe, Talanta
93 (2012) 206.
[9] J. Liu, F. Meng, Z. Li, L. Yu, S. Peng, J. Guo, H. Xu, Biomed. Chromatogr. (2016), DOI:10.1002/bmc.3748.
[10] Z. Zhao, L. Zhang, F. Xue, T. Zhang, Chromatographia 68(9/10) (2008) 731.
[11] G. Marcelin-Jimenez, L. Contreras-Zavala, M. Maggi-Castellanos, A.P. Angeles-Moreno, A. Garcia-
Gonzalez, Bioanal. 4(8) (2012) 909.
[12] M.D. Malesuik, H.M.L. Goncalves, C.S. Paim, E.E.S. Schapoval, M. Steppe, J. Chromatogr. 47(9) (2009)
745.
[13] O.A. Saleh, A.A. El-Azzouny, H.Y. Aboul-Enein, A.M. Badawey, Curr. Pharm. Anal. 10(3) (2014) 215.
[14] M.C. Sharma, Oxid. Commun. 36(4) (2013) 984.
12
[15] Z. Karim, A. Asad, N. Shafi, J. Pharm. Res. 5(2) (2012) 1188.
[16] R. Jain, D.C. Tiwari, P. Karolia, J. Electrochem. Soc. 161(12) (2014) H839.
[17] F.P. Bruno, M.R. Caira, G.A. Monti, D.E. Kassuha, N.R. Sperandeo, J. Mol. Struct. 984(1-3) (2010) 51.
[18] S. Muhu, E. Elamuruguporchelvi, A. Varghese, Spectrochim. Acta A 138 (2015) 743.
[19] B.C. Felix-Sonda, J. Rivera-Islas, D. Herrera-Ruiz, H. Morales-Rojas, H. Hopfl, Cryst. Growth Des. 14(3)
(2014) 1086.
[20] S.S. Sarwade, W.N. Jadhav, B.C. Khade, D. Tayde, Chemica Sinica 6(4) (2015) 1.
[21] D. Tayde, S.S. Sarwade, W.N. Jadhav, B.C. Khade, Archives of Applied Science Research 7(1) (2015) 28.
[22] A.R. Gomes, H. Westh, H. de Lencastre. Antimicrob. Agents Chemother., 50 (2006) 3237.
[23] I.P. Kaur, M. singh, M. Kanwar. Int. J. Pharm., 199 (2000) 119.
[24] A.M. Mansour, Polyhedron 78 (2014) 10.
[25] A.M. Mansour, Dalton Trans. 43 (2014) 15950.
[26] A.M. Mansour, Polyhedron 109 (2016) 99.
[27] M.J. Frisch, G.W. Trucks, H.B. Schlegel, G.E. Scuseria, M.A. Robb, J.R. Cheeseman, V.G. Zakrzewski, J.A.
Montgomery, R.E. Stratmann, J.C. Burant, S. Dapprich, J.M. Millam, A.D. Daniels, K.N. Kudin, M.C. Strain,
O. Farkas, J. Tomasi, V. Barone, M. Cossi, R. Cammi, B. Mennucci, C. Pomelli, C. Adamo, S. Clifford, J.
Ochterski, G.A. Petersson, P.Y. Ayala, Q. Cui, K. Morokuma, D.K. Malick, A.D. Rabuck, K. Raghavachari,
J.B. Foresman, J. Cioslowski, J.V. Ortiz, A.G. Baboul, B.B. Stefanov, G. Liu, A. Liashenko, P. Piskorz, I.
Komaromi, R. Gomperts, R.L. Martin, D.J. Fox, T. Keith, M.A. Al-Laham, C.Y. Peng, A. Nanayakkara, C.
Gonzalez, M. Challacombe, P.M.W. Gill, B. G. Johnson, W. Chen, M.W. Wong, J.L. Andres, M. Head-
Gordon, E.S. Replogle, J.A. Pople, GAUSSIAN 03 (Revision A.9), Gaussian, Inc., Pittsburgh, (2003).
[28] O.R. Shehab, A.M. Mansour, Electroanal., 28 (2016) 1100.
[29] N.T. Abdel-Ghani, A.M. Mansour, Eur. J. Med. Chem., 47 (2012) 399.
[30] National Committee for Clinical Laboratory Standards, NCCLS Approval Standard Document M2-A7,
Vilanova, PA, (2000).
[31] N.T. Abdel-Ghani, A.M. Mansour, J. Mol. Struct. 991 (2011) 108.
[32] A.M. Mansour, Inorg. Chim. Acta 394 (2013) 436.
[33] Z. Trávníček, M. Matiková-MaĬarová, R. Novotná, J. Vančo, K. Štěpánková, P. Suchý, J. Inorg. Biochem.
105 (2011) 937.
[34] R.L. Carlin, R. Burriel, Inorg. Chem. 21 (1982) 4337.
[35] A.Z. El-Sonbati, A.A. El-Bindary, A.F. Shoair, Spectrochim. Acta A, 58 (2002) 3003.
[36] A.K. Singh, V. Balamurugan, R. Mukherjee, Inorg. Chem. 42 (2003) 6497.
[37] C.L. Jain, P.N. Mundley, J. Inorg. Nucl. Chem. 42 (1980) 1769.
[38] S.I. Mostafa, Trans. Met. Chem. 23 (1998) 397.
[39] A. Cotton, G. Wilkinson, Advanced Inorganic Chemistry, second ed., John Wiley and Sons, Inc., Sydney
and Toronto, New York, London, 1972.
[40] M.N. Sokolov, N.E. Fedorova, E.V. Peresypkina, R. Patow, V.E. Fedorov, D. Fenske, Inorg. Chim. Acta
358 (2005) 3914.
[41] F. Weinhold, J.E. Carpenter, The Structure of Small Molecules and Ions, Plenum, New York, 1988, p.
227.
[42] D.C. Onwudiwe, A.C. Ekennia, B.M.S. Mogwase, O.O. Olubiyi, E. Hosten, Inorg. Chim. Acta 450 (2016)
69.
13
O
O
O N+
S O-
HN
N
Na2PdCl4 or K2PtCl4
O
O M
Acetone-H2O/Reflux 2 h Cl
O Cl
O N+
S O- O (1, 2)
HN O
RuCl3.3H2O
N N+
O S O-
O Acetone/Reflux 4 h
HN
Cl N
O
Ru
Cl
Cl OH2
(3)
Scheme 1: Synthesis of Pd(II) (1), Pt(II) (2) and Ru(III) (3) Nitazoxanide complexes.
1.40 2200
µeff vs T
2000
1.35
1/xm vs T 1800
1.30 1600
1400
1.25
1/xm
µeff
1200
1.20
1000
1.15 800
600
1.10
400
1.05 200
50 100 150 200 250 300
T
Fig. 1: The temperature dependence of magnetic susceptibility χm-1 product and μeff of complex 3.
(a)
(b)
(c)
Fig. 2: Local minimum structures of complexes (a) 1, (b) 2, and (c) 3, obtained at the DFT/B3LYP level of
theory.
Fig. 3: TD-DFT/B3LYP/LANL2DZ calculated electronic absorption transitions of complex 2.
NTZ
30
1
2
3
Tetracycline
25
20
% Inhibition (mm/mg)
15
10
0
S. aureus (G+) E. coli (G-)
Fig. 4: Antibacterial activities of NTZ and its complexes and the standard tetracycline against S. aureus as
Gram-positive, and E. coli as Gram-negative bacteria. All experiments were carried out in triplicate and the
mean results are given. DMSO was used as control and its inhibition zone was subtracting from that of
each case
Highlights
• Structural studies of PdII, PtII and RuIII-Nitazoxanide complexes were reported.
• Nitazoxanide prefers to interact with high oxidation state and low-population d-
orbitals Mn+.
• Low-spin octahedral Ru(III) complex with positive Weiss constant was described.
• The releasing of free form of drug in complexes is the key factor of the toxicity.
14
O
O
O N+
S O-
HN
N
Na2PdCl4 or K2PtCl4 O
O M
Acetone-H2O/Reflux 2 h Cl
O Cl
O N+
S O- O (1, 2)
HN O
RuCl3.3H2O
N N+
O S O-
O Acetone/Reflux 4 h
HN
Cl N
O
Ru
Cl
Cl OH2
(3)
Graphical abstract
15
The coordination behavior of the antibacterial drug Nitazoxanide has been investigated against
Pd(II), Pt(II) and Ru(III) ions. Nitazoxanide interacts as a bidentate ligand via the amide C=O
group and thiazole N. Square -planar and low-spin octahedral complexes were reported.
Synopsis
16