Bis(4-nitrobenzenethiolato)tetranitrosyldiiron: synthesis, structure, and pharmacological activity of a new nitric oxide (NO) donor

Article abstract of DOI:10.1007/s11172-017-1944-z

A preparative procedure was developed for the synthesis of new binuclear tetranitrosyl iron complex of the “μ-S” structural type, [Fe₂(SC₆H₄NO₂)₂(NO)₄] (1), comprising the replacement of the thiosulfate ligands in Na₂[Fe₂(S₂O₃)₂(NO)₄]?4H₂O by anions of the 4-nitrobenzenethiolate hydrazinium salt. The salt was obtained by the reduction of bis(4-nitrophenyl) disulfide with hydrazine hydrate in ethanol. The physicochemical properties of the complex in the solid state were studied by IR and M?ssbauer spectroscopy. According to amperometric measurements in aqueous solutions, the complex generates NO in a protic medium without additional photo, thermal, or redox activation, providing prolonged release of NO, since the para-positioned nitro group in the aromatic ligand functions as an additional NO source. A study of the pharmacological activity by the MTT assay showed that the complex possesses a high cytotoxic activity against the MCF7 human breast cancer cells (IC₅₀ = 47 μmol L^–1) and an antibacterial activity against gram-negative (E. coli, BB strain) and gram-positive (M. luteus, 21/26 strain) microorganisms.

Full text of DOI:10.1007/s11172-017-1944-z

Russian Chemical Bulletin, International Edition, Vol. 66, No. 9, pp. 1706—1711, September, 2017
1706
Bis(4ꢀnitrobenzenethiolato)tetranitrosyldiiron: synthesis, structure,
and pharmacological activity of a new nitric oxide (NO) donor*
N. A. Sanina,a,b,c G. I. Kozub,a T. A. Kondrat´eva,a A. A. Terent´ev,a,b,c V. A. Mumyatova,a
P. Yu. Barzilovich,a N. S. Ovanesyan,a and S. M. Aldoshina,c
аInstitute of Problems of Chemical Physics, Russian Academy of Sciences,
1 prosp. Akad. Semenova, 142432 Chernogolovka, Moscow Region, Russian Federation.
Eꢀmail: sanina@icp.ac.ru, alexei@icp.ac.ru
bFaculty of Fundamental Physical and Chemical Engineering,
M. V. Lomonosov Moscow State University,
build. 51, 1 Leninskie gory, 119991 Moscow, Russian Federation.
cMoscow Region State University,
10a ul. Radio, 105005 Moscow, Russian Federation.
A preparative procedure was developed for the synthesis of new binuclear tetranitrosyl
iron complex of the "μꢀS" structural type, [Fe2(SC6H4NO2)2(NO)4] (1), comprising the
replacement of the thiosulfate ligands in Na2[Fe2(S2O3)2(NO)4]•4H2O by anions of the
4ꢀnitrobenzenethiolate hydrazinium salt. The salt was obtained by the reduction of bis(4ꢀ
nitrophenyl) disulfide with hydrazine hydrate in ethanol. The physicochemical properties of
the complex in the solid state were studied by IR and Mössbauer spectroscopy. According to
amperometric measurements in aqueous solutions, the complex generates NO in a protic
medium without additional photo, thermal, or redox activation, providing prolonged release
of NO, since the paraꢀpositioned nitro group in the aromatic ligand functions as an additional
NO source. A study of the pharmacological activity by the MTT assay showed that the
complex possesses a high cytotoxic activity against the MCF7 human breast cancer cells
(IC50 = 47 μmol L–1) and an antibacterial activity against gramꢀnegative (E. coli, BB strain)
and gramꢀpositive (M. luteus, 21/26 strain) microorganisms.
Keywords: nitric oxide, binuclear tetranitrosyl iron complexes, antibacterial activity,
antitumor activity.
Development of methods for the isolation of crystalꢀ
Indeed, for the wild type and mutant E. coli MC4100
strains, it has been shown for the first time9 that the
complex with cysteamine present in physiological conꢀ
centrations controls the culture planktonic growth and
formation of mature biofilms and is comparable in activꢀ
ity with the antibiotic ciprofloxacin. The penicillamine
complex has a vasodilatory activity and can be used in the
NO therapy of the acute myocardial infarction.10 Neutral
complexes containing pyridinꢀ2ꢀyl, pyrimidinꢀ2ꢀyl, phenyl,
and 2ꢀaminophenyl moieties at the sulfur atom have been
shown for the first time to possess high antitumor activity
in vitro,11—13 caused by the induction of apoptosis of
human tumor cells.14
The purpose of the present study is to develop a preꢀ
parative procedure for the synthesis of binuclear tetraniꢀ
trosyl iron complex 1 with a 4ꢀnitrophenylsulfanyl ligand
and to study its properties as a new NO donor in comparꢀ
ison with the previously studied isostructural analogs, genꢀ
erally described as [Fe2(SR)2(NO)4] 1—6.15—19 Comꢀ
plex 1 was studied by spectroscopic techniques, and the
line binuclear nitrosyl iron complexes, synthetic models
of the active sites of the nitrosyl [2Feꢀ2S] ferredoxins,
containing two pharmacophores (NO and RS functional
group) in one molecule, is needed for practical medicine,
as these compounds are promising NOꢀdonors.1 Nitric
oxide is an important signaling molecule with a broad
range of biological activity.2—4 Previously, we successfully
used the functional aliphatic (cysteamine and penicillꢀ
amine) and heterocyclic (hetarylsulfenyl pyridine and
pyrimidine) derivatives as the RSꢀligands for the design
of hybrid nitrosyl iron complexes, because these comꢀ
pounds are structural analogs of the cellular DNA bases
and are widely used to inhibit the growth of malignant
tumors of various origin.5—8 The approach we used to the
design of hybrid structures for the NO chemotherapy of
socially significant diseases came up to our expectations.
* Dedicated to Academician of the Russian Academy of Sciences
G. A. Abakumov on the occasion of his 80th birthday.
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 9, pp. 1706—1711, September, 2017.
1066ꢀ5285/17/6609ꢀ1706 © 2017 Springer Science+Business Media, Inc.

New nitrogen oxide donor
Russ. Chem. Bull., Int. Ed., Vol. 66, No. 9, September, 2017 1707
The Mössbauer absorption spectra were recorded on a WissEl
constant acceleration instrument (Germany). The 57Сo isoꢀ
tope in the Rh matrix served as the γꢀradiation source. The
lowꢀtemperature spectra were measured using a СFꢀ506 heliꢀ
um flow cryostat (Oxford Instruments) with temperature conꢀ
trol. The Mössbauer spectra were processed by the least squares
method with the assumption of Lorentzian shape of unresolved
absorption lines.
effects of the nitro group in the paraꢀposition of the aroꢀ
matic ring of the sulfurꢀcontaining ligand on the NO
donating, cytotoxic, and antibacterial activities of comꢀ
plex 1 were elucidated.
R = 4ꢀO2NC6H4 (1), Ph (2),15 2ꢀH2NC6H4 (3),16 3ꢀO2NC6H4 (4),17
The amount of NO generated by complex 1 in solution was
determined with an amiNOꢀ700 sensing electrode of an inNO
Nitric Oxide Measuring System (Innovative Instruments, Inc.,
USA). The concentration of NO released by the test complex
(0.1 μmol in the sample) was recorded for ∼500 s (with a 0.2 s
step) in a 1% aqueous solution of DMSO. Dimethyl sulfoxide
was purified by a standard procedure.20 The electrochemical
sensor was calibrated against a standard aqueous solution of
NaNO2 (100 μmol L–1), which was added to a mixture of
0.12 М KI and 1 M H2SO4 (2 mL) in water (18 mL). All experꢀ
iments were carried out in aerobic solutions at 25 °С and at
pH 7.0 (pH of solutions was measured with an HI 8314 memꢀ
brane рHꢀmeter (HANNA Instruments, Germany)). The comꢀ
mercial Hydrion buffer (SigmaꢀAldrich, No. 239089) was used.
MTT assay. The cytotoxic activity of the complex was testꢀ
ed against the a human tumor cell line, namely, MCF7 breast
carcinoma cells. The tumor cells were grown in a monolayer in
the RPMI 1640 medium containing 10% fetal calf serum at
37 °С in an atmosphere containing 5% СО2. For the experiꢀ
ments, the cells were seeded into 96ꢀwell plates and grown
under the same conditions. Prior to the experiment, complex 1
was dissolved in DMSO (100 μL) and diluted to a desired conꢀ
centration with the RPMI 1640 culture medium. The final
DMSO concentration in the samples did not exceed 0.1% and
did not affect the cell growth. The cytotoxic activity was deterꢀ
mined by measuring the survival rate of cells in the samples.
The survival rate was determined in the МТТ assay based on
the ability of living cell dehydrogenases to reduce the nonꢀ
colored МТТ reagent to blue formazan crystals soluble in
DMSO.21 Complex 1 was placed into the wells in 20 μL volume
to give final concentrations of 100, 50, and 25 μmol L–1. The
total sample volume was 200 μL. The cells with the compound
were incubated under the aboveꢀindicated conditions for 72 h.
After completion of the incubation, the МТТ reagent was addꢀ
ed and incubation was continued for additional 2 h under the
same conditions. Then the resulting formazan crystals were
dissolved in DMSO (100 μL) at 37 °С for 20 min. The absorꢀ
bance of the resulting solutions was measured at 540 nm on an
optical reader for multiwell plates. The results were expressed
as mean values over four repeated measurements as
(5),18
(6)19
Experimental
Bis(4ꢀnitrobenzenethiolato)tetranitrosyldiiron (1) was synꢀ
thesized from bis(4ꢀnitrophenyl) disulfide (Aldrich) by a known
procedure.16 In the first step, bis(4ꢀnitrophenyl) disulfide (1 g,
3.2 mmol) in ethanol (50 mL) purified by a standard proceꢀ
dure20 was reduced with hydrazine hydrate (SigmaꢀAldrich)
(5 g, 86.2 mmol) with vigorous stirring at room temperature.
After 1 h, the solution turned intensely red; the solvent was
evaporated to leave hydrazinium 4ꢀnitrobenzenethiolate (m.p.
60 °С). In the second step, the thiosulfate ligands of the nitrosyl
complex Na2[Fe2(S2O3)2(NO)4]•4H2O18 were replaced by areꢀ
nethiolate ligands via reductive substitution. For this purpose,
a mixture of Na2[Fe2(S2O3)2(NO)4]•4H2O (0.5 g, 0.9 mmol) in
distilled water (20 mL) and Na2S2O3•5H2O (SigmaꢀAldrich)
(0.5 g) in distilled water (3 mL) was charged into a threeꢀnecked
flask under argon, and a solution of hydrazinium 4ꢀnitrobenzꢀ
enethiolate (1 g, 1.9 mmol) in ethanol (1 mL) was added dropꢀ
wise. The solution thus obtained was stirred for 1 h. The preꢀ
cipitate was collected on a filter under argon and then dissolved
in dichloromethane (special purity grade). The solvent was
evaporated to a minimum amount and the crystals were grown
at 18 °С for 5 days.
Elemental analysis of the polycrystals of complex 1 was
carried out at the Collective Use Analytical Center of the Instiꢀ
tute of Problems of Chemical Physics, RAS, on a Vario Micro
cube CHNS/O analyzer.
For hydrazinium 4ꢀnitrobenzenethiolate, found (%): С, 38.10;
Н, 4.96; N, 22.47; S, 17.52. C6H9N3S1O2. Calculated (%):
C, 38.47; H, 4.84; N, 22.45; S, 17.13.
For complex 1, found (%): C, 26.58; H, 1.56; N, 15.19;
O, 23.37; Fe, 20.28; S, 11.78. Fe2S2C12H8N6O8. Calculated (%):
C, 26.69; H, 1.49; N, 15.55; O, 23.70; Fe, 20.28; S, 11.88.
The IR spectra of complex 1 were measured on a Perkinꢀ
Elmer Spectrum 100X instrument (Collective Use Analytical
Center of the Institute of Problems of Chemical Physics, RAS)
at room temperature with a sensor that collected waves reflectꢀ
ed from the sample surface (independent total internal reflecꢀ
tion technique). The IR spectra were recorded for a crystal of
complex 1 to avoid a damage of the crystal structure, which is
Cell survival rate = (Aexp/Acontrol)•100%.
A compound was considered active if at a concentration of
100 mmol L–1, the cell survival rate for one of three lines was
equal to or less than 50% (IC50 ≤ 100 μmol L–1). The error of
measurements did not exceed 5%.
possible during the traditional grinding with KBr. IR, ν/cm–1
3386.87 (vw), 3098.26 (vw), 1785.79 (vs), 1756.45 (vs), 1597.08 (m),
1575.73 (m), 1514.76 (vs), 1506.75 (vs), 1474.89 (m), 1422.93 (vw),
1397.86 (w), 1363.89 (w), 1340.31 (vs), 1317.08 (m), 1282.05 (w),
1212.15 (vw), 1176.96 (w), 1109.32 (m), 1081.38 (w), 1009.43 (w),
959.70 (vw), 853.51 (s), 840.59 (m), 827.34 (w), 738.17 (s),
681.85(m), 1786 and 1757(νNO).
:
Antibacterial activity. The antibacterial activity of complexꢀ
es 1—6 was measured using serial dilutions by determining the
minimum inhibitory concentration (MIC) that prevents the
visible growth of a bacterium in a broth culture. Successive
twofold dilutions for the test compounds were used. The initial
solutions of the complexes were prepared in DMSO in a maxiꢀ
mum concentration of 20 mmol L–1 and added to 1 mL of the

1708
Russ. Chem. Bull., Int. Ed., Vol. 66, No. 9, September, 2017
Sanina et al.
The 57Fe Mössbauer spectrum of polycrystalline 1 is
a single doublet with the following parameters: quadruꢀ
pole splitting ΔEQ = 1.154(2) mm s–1, isomer shift
LB sterile culture medium (Peptone, 1%; yeast extract, 0.5%;
NaCl, 1%; glucose, 0.1%; pH 6.8—7.0). The final concentraꢀ
tions of the complexes in the samples were from 3.9 μmol L–1
to 2 μmol L–1. The final maximum concentration of DMSO in
the samples was 10%. This DMSO concentration does not affect
the viability of the bacterial agent. Broth culture inoculum with
0.5 density according to the McFarland standard (1 mL) was
added to the test tubes containing solution of complex 1 (the
final concentration of bacteria in the samples was about
5•105 CFU mL–1). The test tubes were incubated in a thermoꢀ
stat at 37 °С, the control samples were incubated over the same
period of time at 4 °С. After 24 h, the results were read out. The
concentration of a compound at which no visible microbial
growth occured in the medium (in comparison with the control
samples) was taken as the MIC. The laboratory gramꢀnegative
E. coli (BB strain) and gramꢀpositive Micrococcus luteus (21/26
strain) cultures were used as bacterial agents. Ampicillin disꢀ
solved in water was used as the reference agent. The range of
δ
= 0.175(1) mm s–1, and absorption linewidth Γ =
Fe
= 0.260(1) mm s–1 (at 85 К) and ΔEQ = 1.129(2) mm s–1
,
δ
Fe = 0.095(1) mm s–1, and Γ = 0.223(1) mm s–1 (at 298 К),
which are similar to those found for the previously studied
"μꢀS"ꢀstructural type complexes.15—20 This attests to the
structural equivalence of the two iron atoms in complex 1
and is consistent with the Xꢀray diffraction data (Fig. 1)
obtained previously for several single crystals of this
compound.22
Like the previously studied neutral binuclear "μꢀS"ꢀ
type tetranitrosyl complexes,14,15,17—19 complex 1 is
readily soluble in DMSO. In a protic medium, complex 1
generates NO without thermal, photo, or enzymatic actiꢀ
vation, and, according to electrochemical data (Fig. 2), it
releases NO in the first seconds after dissolution. The
maximum amount of NO generated by complex 1 by the
8th minute of the experiment is ∼25 nmol per mole of the
complex. Complex 1 has a higher NOꢀdonor activity than
unsubstituted "μꢀS" type complexes with alkyl ligands,
which require additional photoactivation to generate NO
in solution,23 and than the complex with a benzenethiol
residue.15 Furthermore, the kinetic curve illustrating the
time dependence of the amount of NO for complex 1
does not arrive at a plateau ∼1.5 min after the onset of
decomposition under analogous conditions, as in the case
of complex 2 with a benzenethiol residue, and does not
have a clearꢀcut maximum, as in the case of complex 3
with a 2ꢀaminobenzenethiols one,16 for which the NO
release reaches a maximum ∼1 min after the onset of
decomposition and the NO generation is completely
terminated by the 8th minute.
antibiotic concentrations was from 0.49 to 250 μmol L–1
.
Results and Discussion
As the functional RS ligand for the synthesis of nitrosyl
iron complex, we chose 4ꢀnitrophenylsulfanyl, because it
is a representative of benzenethiols containing the pharꢀ
macologically important NO2 group, which readily genꢀ
erates NO, for example, in the presence of redox agents
present in the cells (ascorbate, glutathione, NADP, etc.).
We suggested that combining the SRꢀfunctional ligand with
the NO groups of the structureꢀforming [Fe(SR)2(NO)2]
moiety would ensure a more efficient and longꢀlasting
NOꢀdonating ability of the complex in solutions, which,
in turn, would increase its practical value.
Complex 1 was obtained in two steps according to
Scheme 1.
Analysis of the results of amperometric determination
of NO in solutions of structural analogs of 1 under similar
conditions shows that the introduction of a substituent
into the aromatic ring (phenol or pyridine) of the sulfurꢀ
containing ligand, first, increases the amount of generated
NO and, second, changes the pattern of the kinetic curves
of NO release (Table 1). Complex 1, like complex 5 with
the (5ꢀnitropyridinꢀ2ꢀyl)sulfanyl ligand,19 tends to exhibit
Scheme 1
O(1)
N(1)
O(2)
N(2)
Fe(1)
C(5)
C(1)
C(4)
C(6)
S(1)
C(2)
O(4)
N(3)
O(3)
C(3)
Reagents: i. N2H4•H2O, EtOH; ii. Na2[Fe2(S2O3)2(NO)4]•
•4H2O, Na2S2O3.
Fig. 1. Molecular structure of complex 1 according to Xꢀray
diffraction data.22

New nitrogen oxide donor
Russ. Chem. Bull., Int. Ed., Vol. 66, No. 9, September, 2017 1709
cell apoptosis inducer,12 and for complex 5 (18.8 nmol
per mole of the complex); this is correlated with the data
on the cytotoxic activity of the complexes (Table 2). The
NO/nmol
25
cytotoxic activity of complex 1 (IC50 = 47 μmol L–1
)
20
15
10
5
against MCF7 human breast cancer cells is close to those
of 3 (IC50 = 37 μmol L–1) and 5 (IC50 = 30 μmol L–1
)
when taken at equal concentrations (100 μmol L–1).16
One of the key mechanisms of the NOꢀmediated antiꢀ
microbial action is a chemical change of DNA. It is known
that peroxynitrite and nitrogen dioxide induce cleavage
of DNA strands and formation of adducts;24,25 peroxyꢀ
nitrite also induces peroxidation of bacterial membrane
lipids.26,27 Furthermore, NO inhibits the DNA repair
enzymes. The DNA alkyltransferases contain cysteine resꢀ
idues, which can form S—NO adducts with NO. These
adducts inhibit the transfer of the alkyl group from guaꢀ
nine to the protein. The reactive NO species can modify
proteins not only at cysteine residues, but also at the meꢀ
thionine, tyrosine, phenylalanine, and tryptophan resiꢀ
dues.28 Nitric oxide readily reacts with hemeꢀcontaining
proteins. At high concentrations, reactive nitrogen speꢀ
cies irreversibly bind to the heme, which is hence reꢀ
moved from the protein.29 While NO protects the mamꢀ
malian cells from hydrogen peroxideꢀmediated toxicity,
it sharply increases the hydrogen peroxide toxicity for
bacteria.30 The elimination of iron from metalloenzyme
leads to fast depletion of the iron pool of bacteria. Nitric
oxide induces a decrease in the FeIII concentration, while
accumulation of FeII promotes the formation of hydroxyl
radicals by the Fenton´s reaction.29 The formation of
100
200
300
400
500 τ/s
Fig. 2. Kinetic time dependence for NO generated by nitrosyl
complex 1 in a 1% aqueous anaerobic solution of DMSO
(T = 25 °C, pH 7.0).
prolonged NOꢀdonor activity. The prolonged release of
NO from these two complexes is attributable to the presꢀ
ence of the nitro group in the paraꢀposition of the aroꢀ
matic ligand, which is an additional source of NO. This
assumption could be verified using quantum chemical
calculations and electrospray ionization mass spectromꢀ
etry, because neutral binuclear tetranitrosyl complexes of
this structural type can decompose to give mononuclear
species that generate NO more efficiently.
The amounts of NO released from complex 1 (25 nmol
per mole of the complex) are close to those observed for
complex 2 (20 nmol per mole of the complex), a tumor
Table 1. Amount of NO generated by neutral tetranitrosyl iron complexes 1—6*
Complex
NO release
CNO/nmol per mol of the complex
Kinetic curve pattern
1
2
3
25.0
20.0
40.6
Linearly increases after 4.0 min (prolonged NOꢀdonor activity)
After 1.5 min reaches a constant level (plateau)
After 1.5 min forms a maximum and declines to zero
(a bellꢀshaped curve)
4
5
6
60.0
18.8
4.0
After 5.5 min reaches a constant level (plateau)
Linearly increases after 1.5 min (prolonged NOꢀdonor activity)
After 1.5 min reaches a constant level (plateau)
* Anaerobic 1% aqueous solution of DMSO, pH 7.0, T = 25 °С.
Table 2. Survival of MCF7 human breast cancer cells in the presence of tetranitrosyl iron complexes 1,
3, and 5 and IC50 values for these complexes
Complex
Cell survival rate (%) upon treatment with the complexes
in the concentrations (μmol L–1
IC50/ μmol L–1
)
10
25
50
100
1
3
5
84.86
—
95.28
—
100
—
14.08
24.30
46.08
10.21
12.50
12.61
30
37
47

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Russ. Chem. Bull., Int. Ed., Vol. 66, No. 9, September, 2017
Sanina et al.
Table 3. Antibacterial activity of tetraꢀ
nitrosyl iron complexes 1—6
When tested under aerobic conditions, NO release kinetꢀ
ics may follow other trends, caused by the effect of oxyꢀ
gen, and this requires additional studies.
Complex
MIC/ μmol L–1
E. coli M. luteus
In summary, we developed the first preparative proceꢀ
dure for the synthesis of crystalline nitrosyl iron complex
with a functional sulfurꢀcontaining ligand, the 4ꢀnitroꢀ
benzenethiol residue, which is a structural analog of the
thiopyridine DNA base, and studied this complex by specꢀ
troscopy in the solid phase and in solutions. The use of
the sulfurꢀcontaining ligand with a nitro group in the
paraꢀposition of the benzene ring allows an access to
a nitrosyl iron complex capable of more prolonged NO
generation in aqueous solutions than the NO donor antiꢀ
tumor agents of this class studied previously. Considering
the obtained results on the pharmacological (antitumor
and antibacterial) activity of the test complex and referꢀ
ence complexes, one can state with confidence that the
introduction of functional groups (such as nitro groups)
into the aromatic ring of the sulfurꢀcontaining ligand of
a nitrosyl complex is efficient and provides the design of
new promising antitumor agents of this class of NOꢀdoꢀ
nating systems for NO chemotherapy of infectious disꢀ
eases and cancer.
1
2
3
4
5
6
250
2000
>2000
2000
500
>2000
62.5
125
1000
15
15
31.25
highly reactive free radicals results in the damage of
cellular membranes and DNA.30
The results of the evaluation of the antibacterial activꢀ
ity are summarized in Table 3. The action of ampicillin
used as the reference agent on the bacterial cultures was
in all cases more pronounced than the action of the test
complexes: the MIC of ampicillin for the E. coli culture
was 3.9 μmol L–1 and that for the M. luteus culture was
0.98 μmol L–1. Among the test complexes, the highest
activities against E. coli were found for 1 and 2, the MIC
of which were 250 and 500 μmol L, respectively. The
growth of M. luteus was suppressed by most of the comꢀ
plexes when they were present in concentrations of
15—125 μmol L–1. Generally, all of the test complexes
have a more pronounced action on the M. luteus culture.
A juxtaposition of the NOꢀdonating properties (conꢀ
centration of the generated NO, the pattern of the kinetic
curve) and antibacterial activity of the complexes (see
Table 1 and 3) does not reveal an unambiguous relationꢀ
ship between these two characteristics. Being similar in
the NO amounts generated in the first 500 s, complexes
1, 2, and 5 differ sharply in the antibacterial activity.
Furthermore, complex 1 has a stronger effect on E. coli
than complex 5, while for the M. luteus culture, the oppoꢀ
site relationship holds. Complex 6, which generates NO
in the lowest concentration, has no effect on E. coli;
however, it is one of the strongest growth inhibitors for
M. luteus. Complex 4, which forms NO in the highest
concentration, is also one of the strongest growth inhibiꢀ
tors for M. luteus, but does not affect E. coli. A common
feature of both bacterial cultures is that the bacterial
growth is only slightly inhibited by complex 3; however, this
complex has a broad spectrum of cytotoxic activity.16
Thus, the results demonstrate that apart from the
NOꢀdonating properties of the complexes, the strucꢀ
ture of ligands they contain makes a great contribution to
the antibacterial activity. In particular, complexes 1 and
5 containing ligands with nitro groups affect most effiꢀ
ciently the growth of the E. coli culture, while complexes
with heterocyclic ligands affect most strongly the growth
of M. luteus. It is also worth noting that the NOꢀdonating
properties were evaluated under anaerobic conditions.
The authors are grateful to O. S. Zhukova (N. N.
Blokhin Russian Cancer Research Center, Ministry of
Health of the Russian Federation) for the assay of antituꢀ
mor activity in vitro.
This work was financially supported by the Russian
Foundation for Basic Research (Project No. 17ꢀ03ꢀ
00837).
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Received July 6, 2017