Journal of Inorganic Biochemistry 180 (2018) 204–210

Contents lists available at ScienceDirect

Journal of Inorganic Biochemistry

journal homepage: www.elsevier.com/locate/jinorgbio

Cytotoxic and anticancer properties of new ruthenium polypyridyl T

complexes with different lipophilicities
Anjong Florence Tikuma, Yu Jeong Jeona, Ju Hyun Leea, Min Hee Parkb, In Yeong Baea,b,
⁎ ⁎ ⁎
Sang Heon Kimb,c, , Hye Jin Leed, , Jinheung Kima,
a
Department of Chemistry and Nano Science, Ewha Womans University, Seoul 03760, Republic of Korea
b
Center for Biomaterials, Korea Institute of Science and Technology, Seoul, Republic of Korea
c
Department of Biomedical Engineering, University of Science and Technology, Daejeon, Republic of Korea
d
Department of Chemistry and Green-Nano Materials Research Center, Kyungpook National University, 80 Daehakro, Buk-gu, Daegu 41566, Republic of Korea

A R T I C L E I N F O A B S T R A C T

Keywords: Three ruthenium complexes containing a bidentate piq ligand, [(piq)Ru(bpy)2]2+ (1), [(piq)Ru(phen)2]2+ (2),
Polypyridyl ruthenium complex and [(piq)Ru(DIP)2]2+ (3) (piq = phenylisoquinolinate, bpy = 2,2′-bipyridine, phen = 1,10-phenanthroline,
Cytotoxic activity DIP = 4,7-diphenyl-1,10-phenanthroline), were prepared. The DNA binding properties of complexes 1–3 to
Cellular uptake double-stranded DNA were studied. The binding of 1–3 to calf-thymus DNA (ct-DNA) yielded lower emission
Anticancer activity
intensities than those observed with the corresponding Ru complexes alone. To explore potential interactions of
Cell migration
complexes 1–3 with lipid-rich organs in live cells, the emission properties of the Ru probes were studied with
liposomes. The emission intensities of complexes 1–3 were enhanced to similar extents upon interaction with
liposomes. The cytotoxic activities of the complexes against MDA-MB-231 and HUVECs were evaluated in vitro.
The effects of complexes 1–3 on the survival of MDA-MB-231 cells were examined and compared with that of cis-
platin. Complexes 2 and 3 were more cytotoxic to cancer cells than cis-platin. Complexes 1–3 showed cellular
uptakes of 1.1, 10.6, and 76.6%, respectively, indicating that the greatest amount of complex 3 entered the
cancer cells. Inhibition of cell migration by complexes 1–3 was also evaluated by the wound healing assay.

1. Introduction complexes [13]. The ligand exchange kinetics of metal complexes in

Transition metal complexes constitute a class of chemotherapeutics
that have wide clinical use as antitumor and antiviral drugs. The
structural aspects of metal complexes became a topic of interest after
the discovery of the anticancer properties of cis-platin by Rosenberg
[1]. To date, cis-platin and its analogues are some of the most effective
chemotherapeutic agents in clinical use [2]. cis-Platin is highly effective
for the treatment of testicular and ovarian cancer and is used in com-
bination with other therapies for a variety of other carcinomas, in-
cluding bladder, small cell lung, and head and neck cancers [3].
However, the use of this drug is limited to a small range of tumors
because of primary tumor resistance to cis-platin. In addition, cis-platin
is administered intravenously due to its limited solubility in water, is
associated with severe side effects, and is highly toxic [4].
These limitations have motivated searches for other transition metal
complexes showing beneficial biological properties, wide reactivity
ranges, and lower systemic toxicities [5–12]. These efforts have iden-
tified ruthenium complexes as the most promising transition metal
aqueous solution, which are crucial for anticancer activity and vary for
different metal ions, are very similar for platinum [14] and ruthenium
[14] complexes [15]. Their relatively low toxicity [16] and their ability
to mimic iron in binding to biomolecules have made ruthenium com-
plexes an attractive alternative to platinum-based drugs, since cancer
cells overexpress transferrin receptors to satisfy their increased demand
for iron [17].
We recently studied the interactions between biomolecules and
various metal complexes, including Ru complexes [14], with the aim of
developing cell imaging agents [18–21]. Since the luminescence in-
tensity and cytotoxicity of Ru complexes are strongly ligand-dependent,
considerable research interest has been dedicated to fine-tuning these
properties by changing the commonly used polypyridyl ligands [22].
From this context, we aimed to synthesize three different ruthenium
complexes with a common auxiliary ligand (piq = phenylisoquinoli-
nate) but different chelating ligands (phen = 1,10-phenanthrolinie,
bpy = 2,2′-bipyridine, DIP = 4,7-diphenyl-1,10-phenanthroline). The
bpy complex was reported to have little or no biological activity [23].

⁎
Corresponding authors at: Department of Chemistry and Nano Science, Ewha Womans University, Seoul 120-750, Republic of Korea.
E-mail addresses: skimbrc@kist.re.kr (S.H. Kim), hyejinlee@knu.ac.kr (H.J. Lee), jinheung@ewha.ac.kr (J. Kim).

https://doi.org/10.1016/j.jinorgbio.2018.01.003
Received 24 October 2017; Received in revised form 30 December 2017; Accepted 7 January 2018
Available online 09 January 2018
0162-0134/ © 2018 Elsevier Inc. All rights reserved.
A. Florence Tikum et al.
These complexes vary in lipophilicity, with the DIP ligand complex
bearing the more lipophilic ligand. The aims of this work were two-fold:
1) to investigate the ligand-dependent cytotoxicities of ruthenium
polypyridyl complexes to cancer cells and 2) to determine how the
cytotoxicity, antiproliferation, and cellular uptake by cancer cells are
affected by complex lipophilicity. Three ruthenium complexes con-
taining a common ligand were used for selective interaction with bio-
molecules, followed by cell imaging and other tests. The interaction of
Ru complexes with DNA and liposomes was studied by performing
electronic absorption and emission titrations, cytotoxicity/anti-
proliferation was assessed, and cellular uptake was evaluated using
flow cytometry analysis and fluorescence imaging assays.
2. Experimental section
2.1. Materials
Water was purified with a MilliQ purification system. All reagents
purchased from Aldrich were used without further purification. All
solvents were analytical grade. ct-DNA was purchased from Sigma
(Korea). The metastatic breast cancer cell line MDA-MB-231 (ATCC,
USA) and normal HUVECs (Lonza, USA) were used in this study. The
EZ-CyTox (water-soluble tetrazolium salt (WST)-based cell viability/
cytotoxicity) Assay Kit (Daeil Lab Service, Korea) was used for cell
survival analysis.
2.2. Physical measurement
1
H-NMR spectra were recorded at room temperature on a Bruker
NMR-AL 300 MHz spectrophotometer using DMSO-d6 as the solvent and
TMS as the internal standard. Electrospray mass spectra (ES-MS) were
recorded on an LCQ system. UV/Vis spectra were recorded on a
Hewlett-Packard 8452A diode-array spectrometer with a quartz cuvette
(path length, 1 cm) at room temperature. Emission spectra were re-
corded on a Perkin-Elmer LS 55 luminescence spectrophotometer at
room temperature.
2.3. Synthesis of piq
Ammonium acetate (22.1 mmol) was added to a suspension of
phendione (1.0 mmol) and 4-(4-pyridyl)-benzaldehyde (1.09 mmol) in
acetic acid (25 mL). The reaction mixture was refluxed overnight. After
the mixture was cooled to room temperature, the product was pre-
cipitated. The product was filtered, washed with water and diethyl
ether, and dried under vacuum, yielding a yellow powder [28].
2.4. Synthesis of Ru(bpy)2(piq)2+ (1)
Complex 1 (piq = phenylisoquinolinate, bpy = 2,2′-bipyridine) was
prepared as previously described [23]. Briefly, Ru(bpy)2Cl2 (0.2 mmol)
and piq (0.2 mmol) were dissolved in ethanol and refluxed for 8 h. After
the reaction solution was cooled to room temperature, an excess
amount of KPF6 dissolved in water was added, and a precipitate was
formed. The precipitate was collected by filtration and washed with
water to remove excess KPF6, after which it was purified using column
chromatography on alumina with ethanol/acetonitrile as the eluent.
2.5. Synthesis of Ru(phen)2(piq)2+ (2)
Ru(phen)2Cl2 (0.2 mmol) (phen = 1,10-phenanthroline) and piq
(0.2 mmol) were dissolved in ethanol and refluxed for 8 h. After the
mixture was cooled to room temperature, an excess amount of KPF6
dissolved in water was added, and a precipitate was formed. The solid
was collected by filtration and washed with water to remove excess
KPF6, after which it was purified using column chromatography on
alumina with ethanol/acetonitrile as the eluent. 1H-NMR (DMSO-d6,
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Journal of Inorganic Biochemistry 180 (2018) 204–210
ppm): 7.46 (d, 2H), 7.82 (t, 6H), 8.25 (d, 4H), 8.00 (d, 2H), 8.35 (d,
2H), 8.77 (m, 6H), 8.925 (d, 2H), 9.025 (m, 6H). ESI-MS: m/z 980 [Ru
(phen)2(piq)2+ + PF6−]+, m/z 417.87 [Ru(phen)2(piq)]2+ Anal. Calcd
for RuP2F12N9C48H31: C, 51.25%, H, 2.78%, N, 11.21%. Found: C,
51.95%, H, 2.92%, N, 11.01%.
2.6. Synthesis of Ru(DIP)2(piq)2+ (3)
Complex 3 was synthesized using Ru(DIP)2Cl2 (DIP = 4,7-diphenyl-
1,10-phenanthroline) and piq. 1H-NMR (DMSO-d6, ppm): 7.57 (d, 2H),
7.60 (m, 8H), 7.62 (t, 3H), 7.64 (t, 2H), 7.69 (d, 4H), 7.78 (m, 8H), 7.44
(d, 2H), 8.12 (m, 4H), 8.21 (d, 2H), 8.43 (d, 2H), 9.71 (d, 2H), 9.91 (d,
2H), 9.95 (m, 4H). ESI-MS: m/z 570.13 [Ru(DIP)2(piq)]2+. Anal. Calcd
for RuP2F12N9C72H47: C, 60.51%; H, 3.31%; N, 8.82%. Found: C,
61.01%; H, 3.19%; N, 8.69%.
2.7. DNA binding experiments
Calf thymus DNA (ct-DNA) was prepared as a 1 mM stock solution in
distilled water. Absorption and emission titration experiments were
carried out with Ru complexes at fixed concentrations (10 μM and
2.5 μM, respectively) until the absorbance and emission showed no
further change with increasing ct-DNA concentration. The complex-
DNA solutions were allowed to mix for 10 min at room temperature
before recording.
2.8. Viscosity measurement
Viscosity measurements were carried out using a rheometer main-
tained at 32.04 ± 0.02 °C. The frequency used was 147 Hz, and the
force was 0.5 mPa. DNA samples were prepared for viscosity mea-
surements by sonication in order to minimize complexities arising from
DNA flexibility. Flow time was measured at least five times for each
sample, after which the average flow time was calculated. Data are
presented as (η/η0)1/3 versus [Ru]/[DNA], where η is the viscosity of
DNA in the presence of the Ru(II) complex, and η0 is the viscosity of the
DNA solution alone.
2.9. Liposomes binding experiment
DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine, Avanti Lipids)
was solubilized in CHCl3 and the excess solvent was evaporated in a
vacuum oven at 37 °C for 2 h. The film was then rehydrated with so-
dium phosphate buffer (pH 7.4) to a concentration of 50 mM. This
procedure was followed by 3 cycles of annealing at 60 °C, followed by
vortexing. Each step was 3 min in duration. A heterogeneous suspension
of liposomes was obtained, which was incubated overnight at 4 °C. A
homogenized liposome suspension was obtained by sonicating the
suspension for 5 min before extrusion through a 0.22 μm filter mem-
brane [29]. Solutions of 1, 2, and 3 (2.5 μM, 0.1% CH3CN: 99.9% SPB)
were then incubated with liposomes (0–15 μg/mL in SPB), after which
the emission spectra were collected.
2.10. Cell culture
MDA-MB-231 cells were maintained in RPMI 1640 medium with
10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. NIH3T3
cells were cultured in a 5% CO2 atmosphere at 37 °C in high-glucose
DMEM with 10% FBS and 1% penicillin/streptomycin. The medium
was replaced every 2 days.
2.11. Cell survival experiments
Cells were seeded in a 96-well plate (7000 cells per well) supple-
mented with culture medium and then incubated in a 5% CO2 atmo-
sphere at 37 °C. After 24 h, the culture medium was replaced with serial
A. Florence Tikum et al. Journal of Inorganic Biochemistry 180 (2018) 204–210

N H
N N N
Ru N
N N N
N
2 PF6 N
1 N
H
N N
Ru N
N N
N
N
N 2 PF6
N
H 3
N N
Ru N
N N
N
N
2 PF6 2

Fig. 1. Chemical structures of Ru complexes 1, 2, and 3.

(a) (a)
1.5
Absorbance

1.0
Absor bance (a.u.)

0.5 0.5
1 3
2
0.0
200 400 600 800
W avelength(nm )
(b) 18
16
0.0
14 1
Intensity (a.u.)

200 400 600 800

12 2
10 W avelength(nm )
8 (b)
3 14
6
4
12
2
0 10
500 550 600 650
Intensity (a.u.)

W avelength(nm ) 8

Fig. 2. (a) Absorption spectra of 10 μM 1 (black dotted line), 2 (red dashed line), and 3 6
(blue solid line) in 50 mM phosphate buffer (pH 7.4). The concentration of each complex
is 2.5 μM. (b) Emission spectra of complexes 10 μM 1 (black dotted line), 2 (red dashed 4
line), and 3 (blue solid line) in 50 mM phosphate buffer (pH 7.4). The concentration of
each complex is 2.5 μM. Excitation wavelength = 450 nm. (For interpretation of the re- 2
ferences to color in this figure legend, the reader is referred to the web version of this
article.) 0
500 550 600 650 700

dilutions of complexes in fresh medium (6.25, 12.5, 25, 50, 100, and
200 μM), after which the cells were incubated for another 48 h. For the
cell survival assay, cells were washed with PBS, and then cell survival
was assessed using a CCK-8 Assay Kit according to the manufacturer's
protocol. Untreated and cis-platin-treated cells were used for compar-
ison.
2.12. Fluorescence imaging assay
MDA-MB-231 cells and HUVECs were seeded in 96-well plates at a
density of 10,000 cells per well and incubated for 24 h. Then, the
W avelength(nm )
Fig. 3. (a) Absorption spectra of complex 2 at 10 μM upon the addition of calf-thymus
DNA (0–30 μM) in 50 mM phosphate buffer (pH 7.4). (b) Emission spectra of complex 2 at
2.5 μM upon treatment with calf-thymus DNA (0–30 μM) in 50 mM phosphate buffer
(pH 7.4). Excitation wavelength = 450 nm.
culture medium was replaced with a Ru complex solution (10−5 M),
and the cells were incubated for another 72 h. After washing with PBS,
cells were fixed using para-formaldehyde solution and then viewed
under an LSM 700 laser scanning confocal microscope (Zeiss,
Germany).

206
A. Florence Tikum et al.
2.13. Flow cytometric analysis
MDA-MB-231 cells and HUVECs were seeded in 6-well plates at a
density of 50,000 cells per well and incubated for 24 h. After the cells
were washed twice with PBS and trypsinized for 3 min, the cells were
harvested and washed twice with PBS. The cells were fixed in para-
formaldehyde solution and analyzed on a Guava easyCyte™ flow cyt-
ometer (Merck Millipore, Germany).
2.14. Statistical analysis
Quantitative data are expressed as mean ± standard deviation. The
significance of differences in the mean values was evaluated using
Student's t-test. Differences were considered significant when the P
value was less than .01 or .05, as indicated.
2.15. Wound healing assay
MDA-MB-231 breast cancer cells were seeded in 6-well plates at a
density of 180,000 cells per well and incubated for 48 h to form
monolayers. The monolayers were wounded by scratching with a
plastic tip. The wounded monolayers were then washed several times
with PBS to remove cell debris and incubated in medium with Ru
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Journal of Inorganic Biochemistry 180 (2018) 204–210
Fig. 4. Cell survival of Ru complexes and cis-platin against
MDA-MB-231 at different time intervals. Cells were treated
with 1–3 at increasing concentration (6.25, 12.5, 25, 50,
100, and 200 μM) and untreated cells were used as the
control. Cell survival was analyzed by the CCK-8 Assay Kit.
complexes (25 μM) for 48 h. Cell migration into the wound gap and the
migration distance were assessed under an inverted microscope at
various time points after wounding. Ru complexes, which alter cell
mobility and growth, can decrease or increase the rate of wound
healing.
3. Results and discussion
3.1. Synthesis and characterization
Three ruthenium complexes bearing a common piq ligand, [(piq)Ru
(bpy)2]2+ (1), [(piq)Ru(phen)2]2+ (2), and [(piq)Ru(DIP)2]2+ (3)
(piq = phenylisoquinolinate, bpy = 2,2′-bipyridine, phen = 1,10-phe-
nanthroline, DIP = 4,7-diphenyl-1,10-phenanthroline), were prepared
(Fig. 1). Complex 1 was prepared according to known synthesis
methods [23]. Complexes 2 and 3 were synthesized by reacting Ru
(phen)2Cl2 and Ru(DIP)2Cl2, respectively, with equimolar amounts of
piq. The desired ruthenium complexes were isolated as PF6− salts and
purified by column chromatography. All Ru complexes were char-
acterized by ESI-MS, 1H-NMR, and elemental analysis. These complexes
were soluble in CH3CN and DMSO, but had low solubility in water.
Complex 1 had absorption bands at 288, 328, and 458 nm; 2 at 262,
287, and 450 nm; and 3 at 292, 357, and 486 nm in 50 mM phosphate
A. Florence Tikum et al. Journal of Inorganic Biochemistry 180 (2018) 204–210

/&#/$ *78'%




PQPG  
4W
DR[
RKS  
4W
&+2
RKS  
4W
RJGP
RKS  

Fig. 5. Cellular uptake of complexes 1–3 (25 μM) into (a) cancer cell line (MDA-MB-231) and human normal cells (HUVEC) studied by laser scanning confocal microscope for 72 h. The
scale bar is 200 μm. (c) Flow cytometric histograms and percent of internalized Ru complex are shown in the table. Cellular uptake was analyzed by guava easyCyte™ Flow Cytometers
(Merck Millipore, Germany).

buffer (pH 7.4) (Fig. 2a). The bands at 288 and 326 nm of 1, 265 and
287 nm of 2, and 292 and 357 nm were derived from intraligand π-π*
electronic transitions. The bands at 458 nm of complex 1, 450 nm of 2,
and 486 nm of 3 were attributed to metal-to-ligand charge transfer
transitions. The luminescence spectra of the Ru complexes were also
obtained. The CH3CN solutions of these Ru complexes were almost non-
emissive, while the aqueous solutions were strongly luminescent. Upon
excitation at 450 nm, complexes 2 and 3 emitted light at 593 and
609 nm, respectively. Complex 1 emitted light at 598 nm (Fig. 2b).
3.2. DNA binding properties
Interest in poly-pyridyl Ru complexes was generated by their in-
teresting emission properties upon interaction with double-stranded
DNA. These properties led us to investigate the interaction between
DNA and poly-pyridyl Ru ruthenium complexes by measuring their
absorption, luminescence, and viscosity. Absorption and luminescence
titrations were carried out in 50 mM sodium phosphate buffer (pH 7.4).
The resulting mixtures were incubated for 10 min before recording
absorption spectra. As shown in Fig. 3a, some changes in the absorption

208
A. Florence Tikum et al.
bands were observed upon the addition of calf thymus DNA (ct-DNA) to
2, indicating that there is an interaction between 2 and DNA. Com-
plexes 1 and 3 also exhibited some increases in the absorption spectra
upon addition of ct-DNA (Fig. S1). Then, luminescence spectra were
obtained to further understand the interaction of complexes 1–3 with
ct-DNA. In contrast to previously described polypyridyl complexes
[24–27], the emission band of 2 was quenched as the DNA concentra-
tion increased (Fig. 3b). Complexes 1 and 3 also yielded similar
quenching results, confirming that they also interacted with ct-DNA
(Fig. S2). However, complex 3 showed difference that the intensity
changes were relatively smaller than those with complexes 1 and 2,
upon treatment with ct-DNA. To investigate the binding mode of these
complexes, viscosity measurements were carried out using ethidium
bromide as a control (Fig. S3). In contrast to ethidium bromide, the
viscosities obtained with complexes 1–3 did not increase with in-
creasing Ru complex concentration, indicating that these complexes
1–3 interact with ct-DNA in a non-intercalative mode.
3.3. Interactions of the Ru complexes with liposomes
Lipids provide a hydrophobic environment by forming a double
layered membrane present in many biological systems. The emission
changes of complexes 1–3 were examined after addition of liposomes in
order to investigate whether the Ru complexes interact with hydro-
phobic membranes. Complex 1 showed a significant increase in MLCT
luminescence around 598 nm after treatment with liposomes (Fig. S4).
The addition of liposomes to complexes 2 and 3 also produced an in-
crease in luminescence at 593 and 609 nm, respectively; these en-
hancements were similar to that observed with complex 1. Based on the
well-reported photophysical properties of polypyridyl Ru complexes,
the hydrophobic interaction between complex 1 and liposomes shielded
the Ru complexes from water, thereby enhancing emission [17–18].
3.4. Cell survival assay in vitro
The effects of Ru complexes 1–3 and cis-platin on the survival of
MDA-MB 231 cells were assessed in vitro. Cells were exposed to the Ru
complexes or cis-platin for 48 h (6.25, 12.5, 25, 50, 100, or 200 μM),
after which viability was assessed using a CCK-8 Assay Kit. cis-Platin is
one of the most effective drugs for treating ovarian, testicular, and head
209
Journal of Inorganic Biochemistry 180 (2018) 204–210
Fig. 6. Temporal progress of the wound healing in MDA-MB-
231 treated with 25 μM of complexes 1–3 at 48 h. The scale bar
is 500 μm.
and neck carcinomas. Thus, untreated and cis-platin-treated cells were
used for comparison. The survival rates of cis-platin-treated and com-
plex 1-treated cells after 48 h were similar. However, the survival rates
of complex 2-treated and complex 3-treated cells were lower than those
of cis-platin-treated and complex 1-treated cells regardless of con-
centration, indicating that complexes 2 and 3 are more cytotoxic to
cancer cells than cis-platin (Fig. 4).
3.5. Cellular uptake
To compare the penetration of the Ru complexes into cancer cells
and normal human cells, the two types of cells were incubated with
complexes 1–3 (25 μM solutions) for 72 h. Fluorescence microscopy
(Fig. 5a and b) and flow cytometric analysis (Fig. 5c) indicated that
complexes 1–3 showed different penetration trends into cells. Complex
3, which harbors extra phenyl rings, appeared to have enhanced pe-
netration of cancer cells compared with complexes 1 and 2. All three
complexes showed higher cellular uptake by cancer cells than normal
cells, indicating that complexes 1–3 could selectively accumulate in
cancer cells but not in normal cells. Flow cytometry analysis revealed
that complexes 1–3 showed efficient penetration into cancer cells. Cu-
mulatively, these results indicate that complex 3 penetrates cancer cells
better than the other two complexes. This finding is in line with the fact
that the more lipophilic is a compound, the easier it is to penetrate cells.
Moreover, high magnification fluorescence images revealed different
penetration efficiencies into cancer cells and normal human cells, i.e.,
complexes 1–3 were preferentially taken up by cancer cells (Fig. 5c).
3.6. Wound healing assay
The motility of MDA-MB-231 cells, a commonly used breast cancer
cell line, was examined by measuring their metastatic potential.
Confluent cell monolayers were scratched to form wounds and cultured
in the presence of various concentrations of complexes 1–3 (25 μM) for
48 h. Treatment of MDA-MB-231 with complex 1 led to a concentration-
dependent decrease in wound healing (i.e., cell migration) (Figs. 6 and
S5). Cells treated with complexes 2 and 3 displayed cell debris ag-
gregates (shown in orange), implying that these complexes potentially
stop the growth and spread of cancer cells and merit further in-
vestigation in the future.
A. Florence Tikum et al.
4. Conclusion
We designed and synthesized three ruthenium polypyridyl com-
plexes with the same auxiliary structure but different chelating ligands.
These complexes showed different lipophilicities. The DNA binding,
viscosity, cytotoxicity, cellular uptake, and migration of each of the Ru
complexes were investigated. All three complexes were observed to
bind strongly to calf thymus DNA via a non-intercalative binding mode,
as evidenced by fluorescence and viscosity measurements. Of the three
complexes, complex 3 was the most lipophilic and also exhibited the
highest cytotoxicity and the greatest uptake into cancer cells. These
attributes might result from its high lipophilicity. Our complexes se-
lectively accumulated in cancer cells compared to normal cells and
were demonstrated to possess greater cytotoxicity and metastatic ac-
tivity than cis-platin. From these experimental data, we infer that the
anticancer activities of these complexes are significantly ligand-de-
pendent and also that complex lipophilicity enhances cellular uptake
and anticancer activity. We anticipate that our findings will contribute
to the search, modification, and design of new and more effective anti-
cancer drugs.
Acknowledgements
This work was supported by the National Research Foundation of
Korea (NRF) grant funded by the Korea government (MSIP) (No. NRF-
2017R1A5A1015365 and NRF-2016R1A2B4012488, to J. Kim) and by
a grant from the Ministry of Science, ICT, and Future Planning (NRF-
2016R1A284012026, to H. Lee).
Appendix A. Supplementary data
See Supplementary Information for additional data, such as ab-
sorption and luminescence spectra, viscosity and wound healing results.
This material is found online at https://doi.org/10.1016/j.jinorgbio.
2018.01.003.
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