European Journal of Medicinal Chemistry 170 (2019) 195e202

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European Journal of Medicinal Chemistry

journal homepage: http://www.elsevier.com/locate/ejmech

Research paper

In vitro and in vivo antitumor activities of three novel binuclear

platinum(II) complexes with 40 -substituted-2,20 :60 ,200 -terpyridine
ligands
Qi-Pin Qin a, *, 1, Zhen-Feng Wang a, 1, Shu-Long Wang a, Dong-Mei Luo a, Bi-Qun Zou b, d,
Peng-Fei Yao c, Ming-Xiong Tan a, Hong Liang d, **
a
Guangxi Key Laboratory of Agricultural Resources, Chemistry and Biotechnology, College of Chemistry and Food Science, Yulin Normal University, 1303
Jiaoyudong Road, Yulin, 537000, China
b
Department of Chemistry, Guilin Normal College, 9 Feihu Road, Gulin, 541001, China
c
Guangxi Colleges and Universities Key Laboratory of Regional Ecological Environment Analysis and Pollution Control of West Guangxi, College of
Chemistry and Environmental Engineering, Baise University, Baise, Guangxi, 533000, China
d
State Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, School of Chemistry and Pharmacy, Guangxi Normal University,
15 Yucai Road, Guilin, 541004, China

a r t i c l e i n f o a b s t r a c t

Article history: Herein, we report the design and synthesis of three novel binuclear platinum(II) complexes, [Pt(tpbtpy)
Received 4 January 2019 Cl][Pt(DMSO)Cl3] (tpbtpy-Pt), [Pt(dthbtpy)Cl][Pt(DMSO)Cl3],CH3OH (dthbtpy-Pt), and [Pt(qlbtpy)Cl]
Received in revised form [Pt(DMSO)Cl3],CH3OH (qlbtpy-Pt) with 40 -(3-thiophenecarboxaldehyde)-2,20 :60 ,200 -terpyridine (tpbtpy),
2 March 2019
40 -(3,5-bis (1,1-dimethylethyl)-2-hydroxy-benzaldehyde)-2,20 :60 ,200 -terpyridine (dthbtpy) and 40 -(2-
Accepted 5 March 2019
Available online 9 March 2019
quinolinecarboxaldehyde)-2,20 :60 ,200 -terpyridine (qlbtpy) as ligands, respectively. All three novel binu-
clear platinum(II) complexes tpbtpy-Pt, dthbtpy-Pt, and qlbtpy-Pt were characterized by single-crystal
X-ray diffraction analysis, spectroscopic analysis (ESI-MS, IR, 1H NMR), and elemental analysis. Addi-
Keywords:
40 -(3-thiophenecarboxaldehyde)-2,20 :60 ,200 ;-
tionally, the cytotoxicity of tpbtpy-Pt, dthbtpy-Pt and qlbtpy-Pt was assessed with human non-small
terpyridine cell lung cancer cell line (NCIeH460 cells), yielding IC50 values in the range of 0.35e12.09 mM with
Platinum(II) complexes tpbtpy-Pt as the most potent and qlbtpy-Pt as the least potent complexes. Mechanistic studies indicated
Cell apoptosis that tpbtpy-Pt and dthbtpy-Pt induced apoptosis through mitochondrial dysfunction and telomerase
Telomerase inhibition inhibition. In a NCIeH460 xenograft model, when administered at 10.0 mg kg1 every 2 days, tpbtpy-Pt
Dysfunction of mitochondria was shown to significantly reduce tumor growth (tumor growth inhibition rate (IR) ¼ 70.1%, p < 0.05).
Therefore, tpbtpy-Pt is a promising Pt(II) complex for further translational studies and clinical evaluation
as an antitumor agent.
© 2019 Elsevier Masson SAS. All rights reserved.

1. Introduction complexes [3e10], such as platinum(IV)-benzhydrazide bis-conju-

Pt(II)-based drugs are widely used in the clinical treatment of
lung, testicular (NT2), and ovarian cancers. [1e8] However, high
drug toxicity and drug resistance limit their clinical utility, and thus
researchers are encouraged to design and synthesize new Pt(II/IV)
* Corresponding author.
** Corresponding author.
E-mail addresses: qpqin2018@126.com (Q.-P. Qin), shulonghs@163.com
(S.-L. Wang), zoubiqun@163.com (B.-Q. Zou), yaopengfei1986@163.com (P.-F. Yao),
hliang@gxnu.edu.cn (H. Liang).
1
These authors contributed equally to this work.
gate, organometallic Pt(II) complexes [Pt(C^N)Cl(DMSO)] (C^N ¼
N,N-dimethyl-1-(2-aryl)methanamine- k2C2,N), Pt(IV) prodrugs,
cationic polynuclear Pt(II) complexes, and others. [1e12].
In addition, a large number of Pd(II), Fe(II/III), Pt(II), Zn(II), Ir(III),
Cu(II), Au(III), Ru(II/III), V(III), and Os(III) terpyridyl complexes have
been designed and studied against cancer cells, and these com-
pounds showed better antitumor activity than the corresponding
terpyridyl ligands [13e43]. For instance, Che described a series of
cyclometalated platinum(II) complexes, [(C^N^N)PtII(N,N0 -n-
Bu2NHC)]PF6 (HC^N^N ¼ 6-phenyl-2,20 -bipyridine), that are in-
hibitors of apoptosis (IAP) in vitro and in vivo [37]. Hussain
investigated mixed-ligand copper(II) complex, [Cu(Fc-tpy)(CQ)]

https://doi.org/10.1016/j.ejmech.2019.03.014
0223-5234/© 2019 Elsevier Masson SAS. All rights reserved.
196 Q.-P. Qin et al. / European Journal of Medicinal Chemistry 170 (2019) 195e202

NO3 (CQ ¼ 5-chloro-7-iodo-8-hydroxyquinoline), which is mainly

accumulated in the mitochondria [39]. Lippard and co-authors re-
ported that the cationic complexes [OsCl2N(tpy)]Cl
(tpy ¼ 2,2':60 ,200 -terpyridine) and [OsCl3N(phen)] (phen ¼ o-phe-
nanthroline) had activity against glioblastoma (GBM) initiating
tumor cells in vitro and in vivo [38]. Shul'pin and Rahiman sug-
gested that 40 -(3-thiophenecarboxaldehyde)-2,20 :60 ,200 -terpyridine
(tpbtpy) copper(II) complexes [CuCl2(tpbtpy)] and [Cu(tpbtpy)2]Cl2
could induce HCT116 and MCF-7 cell apoptosis much more effec-
tively than other similar metal complexes [42,43]. However, no 40 -
(3-thiophenecarboxaldehyde)-2,20 :60 ,200 -terpyridine (tpbtpy) Pt(II)
complex has been reported in the literature, and the detailed
in vitro and in vivo anticancer mechanisms of 40 -(3-
thiophenecarboxaldehyde)-2,20 :60 ,200 -terpyridine (tpbtpy) plati-
num(II) complexes remain unexplored.
Herein, we report the design, synthesis, and characterization of
three novel binuclear platinum(II) complexes, [Pt(tpbtpy)Cl] Fig. 1. ORTEP view of tpbtpy-Pt.
[Pt(DMSO)Cl3] (tpbtpy-Pt), [Pt(dthbtpy)Cl][Pt(DMSO)Cl3],CH3OH
(dthbtpy-Pt) and [Pt(qlbtpy)Cl] [Pt(DMSO)Cl3],CH3OH (qlbtpy-Pt)
with 40 -(3-thiophenecarboxaldehyde)- 2,20 :60 ,200 -terpyridine
(tpbtpy), 40 -(3,5-bis(1,1-dimethylethyl)-2-hydroxy- benzalde-
hyde)-2,20 :60 ,200 -terpyridine (dthbtpy) and 40 -(2-
quinolinecarboxaldehyde) 2,20 :60 ,200 -terpyridine (qlbtpy) as li-
gands, respectively (Scheme 1). In addition, the effects of the three
novel binuclear platinum(II) complexes, tpbtpy-Pt, dthbtpy-Pt and
qlbtpy-Pt, on cell proliferation and apoptosis in vitro and in vivo
were evaluated.

2. Results and discussion

2.1. Synthesis and characterization

The synthesis of 40 -(3-thiophenecarboxaldehyde)-2,20 :60 ,200 -

terpyridine (tpbtpy), 40 -(3,5-bis(1,1-dimethylethyl)-2-hydroxy-
benzaldehyde)-2,20 :60 ,200 -terpyridine (dthbtpy), and 40 -(2-
quinolinecarboxaldehyde)-2,20 :60 ,200 -terpyridine (qlbtpy) were
carried out as reported by Zou et al. (Scheme 1) [40e43]. Briefly, Fig. 2. ORTEP view of dthbtpy-Pt. The solvent CH3OH is omitted for clarity.
[Pt(tpbtpy)Cl][Pt(DMSO)Cl3] (tpbtpy-Pt), [Pt(dthbtpy)Cl]
[Pt(DMSO)Cl3],CH3OH (dthbtpy-Pt) and [Pt(qlbtpy)Cl][Pt(DMSO)
Cl3],CH3OH (qlbtpy-Pt) were synthesized by combining cis-
Pt(DMSO)2Cl2 (2.0 mmol) and tpbtpy, dthbtpy or qlbtpy (1.0 mmol)
in CH3OH (25.0 mL) and DMSO (0.2 mL) at 60  C for 6.0 h. All three
novel binuclear platinum(II) complexes tpbtpy-Pt, dthbtpy-Pt and
qlbtpy-Pt were characterized by single-crystal X-ray diffraction
analysis (Figs.13 and Tables S1S9), spectroscopic analyses (ESI-
MS, IR, 1H NMR), and elemental analysis (Fig. S1S9). Furthermore,
we showed that tpbtpy-Pt, dthbtpy-Pt and qlbtpy-Pt
(2.0  105 mol/L) were stable for 48 h in Tris-HCl buffer solution by
ESI-MS (Figs. 2, 4 and 6) [44e51].

Fig. 3. ORTEP view of qlbtpy-Pt. The solvent CH3OH is omitted for clarity.

2.2. Crystal structures

Scheme 1. Synthetic route of the syntheses of tpbtpy, dthbtpy, qlbtpy and their three
novel binuclear platinum(II) complexes tpbtpy-Pt, dthbtpy-Pt and qlbtpy-Pt. Re- The crystal structures of the three novel binuclear platinum(II)
agents and solvents are as follows: (a) ethanol, 25% NH3,H2O (aq), KOH; (b) cis- complexes tpbtpy-Pt, dthbtpy-Pt and qlbtpy-Pt were determined
Pt(DMSO)2Cl2 (2.0 mmol), 25.0 mL CH3OH and 0.2 mL DMSO, 60  C, 6.0 h.
 Q.-P. Qin et al. / European Journal of Medicinal Chemistry 170 (2019) 195e202 197

by X-ray crystallography, as shown in Figs.13. A list of angels ( )
and selected bond lengths (Å) is shown in Tables S1S9, which
were within the expected range [44e51]. The tpbtpy-Pt, dthbtpy-
Pt or qlbtpy-Pt complex consists of one cationic [Pt(tpbtpy)Cl]þ,
[Pt(dthbtpy)Cl]þ or [Pt(qlbtpy)Cl]þ, and one anionic [Pt(DMSO)
Cl3], where the two central Pt(II) atoms were arranged in a four-
coordinated distorted square planar geometry.
2.3. In vitro cytotoxicity
We studied the cytotoxicity of tpbtpy, dthbtpy, qlbtpy, cis-
Pt(DMSO)2Cl2, three novel binuclear platinum(II) complexes
tpbtpy-Pt, dthbtpy-Pt, qlbtpy-Pt, and cisplatin at various con-
centrations (0.325, 0.75, 1.25, 2.5, 5.0, 10.0 and 20.0 mM) against
human NCIeH460 (non-small cell lung cancer cells), T-24 (bladder
cancer cells), SK-OV-3 (ovarian cancer cells), A549 (lung carcinoma
cancer cells), and human liver Hl-7702 normal cells. The IC50 values
was obtained by the MTT assay after exposure to tpbtpy, dthbtpy,
qlbtpy, cis-Pt(DMSO)2Cl2, tpbtpy-Pt, cisplatin, dthbtpy-Pt, and
qlbtpy-Pt for 48 h (Table 1). It was evident that tpbtpy-Pt con-
taining the 40 -(3-thiophenecarboxaldehyde)-2,20 :60 ,200 -terpyridine
(tpbtpy) ligand demonstrated the greatest anticancer activity
(IC50 ¼ 0.35 ± 0.08 mM) against human NCIeH460 cells, a non-small
cell lung cancer cell line. The antiproliferative activity of the three
novel binuclear platinum(II) complexes tpbtpy-Pt, dthbtpy-Pt,
qlbtpy-Pt, and cisplatin increased in the order of tpbtpy-
Pt > dthbtpy-Pt > qlbtpy-Pt > cisplatin, which may be due to the
different substituent group (tpbtpy > dthbtpy > qlbtpy) will affect
the planarity of the angle among the pyridine ring planes in three
novel binuclear Pt(II) complexes, especially 3-
thiophenecarboxaldehyde substitution in the tpbtpy ligand of the
tpbtpy-Pt complex, very similar to the results obtained by Sadler
et al. [52e55]. Notably, the three novel binuclear platinum(II)
complexes tpbtpy-Pt, dthbtpy-Pt, and qlbtpy-Pt displayed higher
antiproliferative activity against the human non-small cell lung
cancer cell line (NCIeH460 cells) as compared to the human liver
HL-7702 normal cells (Table 1).
lung cancer cell line (NCIeH460 cells). In NCIeH460 cells, upon
treatment with tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) for
24 h, down-regulation of c-myc and hTERT was observed compared
with control, especially with tpbtpy-Pt (0.35 mM) treated cells
(Fig. 4B and C). Furthermore, telomerase activity was decreased after
treatment with tpbtpy-Pt (0.35 mM, inhibitory rates of 62.50%)) and
dthbtpy-Pt (5.53 mM, inhibitory rates of 52.25%)) at 24 h (Fig. 4A),
which may be related to the inhibition of c-myc and hTERT (Fig. 4)
[47,56e67]. Finally, G1 accumulation increased rapidly at 24 h
(Fig. 5AeC), and the G1 phase populations observed in the tpbtpy-Pt
(0.35 mM) and dthbtpy-Pt (5.53 mM) groups were 69.06% and
66.73%, respectively, and cyclin D1 and CDK2 were down-regulated
at 24 h in NCIeH460 cells (Fig. 5D and E). At the same time, the
quinone oxidoreductase isozyme I (NQO1) Pt(IV) complexes caused
arrest at S phase which was different from that of tpbtpy-Pt
(0.35 mM) and dthbtpy-Pt (5.53 mM), demonstrating that the anti-
cancer mechanism of tpbtpy-Pt (0.35 mM) and dthbtpy-Pt
(5.53 mM) could distinguish from NQO1 Pt(IV) complexes. In general,
this result suggested that inhibition of telomerase activity by
tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) in NCIeH460 cancer
cells led to partial growth arrest [47,56e67].
2.5. Dysfunction of mitochondria
It is well-known that mitochondrial changes, including in-
creases in ROS generation, loss of DJm (mitochondrial membrane

2.4. Inhibition of telomerase activity and the related proteins by

tpbtpy-Pt and dthbtpy-Pt in NCIeH460 cells

A large body of literature has reported that inhibition of telo-

merase activity by chemical complexes or compounds leads to the
decrease in the level of hTERT and c-myc proteins [56e66], conse-
quently inducing tumor cell cycle arrest and down-regulating
related-proteins [47,56e67]. Thus, we next studied whether two
novel binuclear platinum(II) complexes tpbtpy-Pt (0.35 mM) and
Fig. 4. Modulation of telomerase activity by tpbtpy-Pt (0.35 mM) and dthbtpy-Pt
dthbtpy-Pt (5.53 mM) could down-regulate related-proteins, (5.53 mM) in NCIeH460 cancer cells. (A) Analysis of telomerase activity. (B and C) c-myc
including c-myc, cyclin D1, hTERT and CDK2, inhibit telomerase ac- and hTRET proteins in tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) treated cells
tivity, and induce tumor cell cycle arrest in a human non-small cell were detected by Western blot.

Table 1
The IC50 values (mM) of tpbtpy, dthbtpy, qlbtpy, cis-Pt(DMSO)2Cl2, three novel binuclear platinum(II) complexes tpbtpy-Pt, dthbtpy-Pt, qlbtpy-Pt, and cisplatin towards
human NCIeH460 (non-small cell lung cancer cells), T-24 (bladder cancer cells), SK-OV-3 (ovarian cancer cells), A549 (lung carcinoma cancer cells) and human liver Hl-7702
normal cells after incubation for 48 h.

Compound NCIeH460 SK-OV-3 T-24 A549 HL-7702

tpbtpy 25.41 ± 0.81 34.83 ± 1.08 19.61 ± 0.69 14.21 ± 0.19 38.22 ± 0.66
tpbtpy-Pt 0.35 ± 0.08 4.52 ± 0.29 1.24 ± 0.55 6.03 ± 1.22 40.22 ± 1.58
dthbtpy 30.11 ± 0.56 35.22 ± 2.01 20.88 ± 0.44 16.31 ± 0.66 35.11 ± 1.69
dthbtpy-Pt 5.53 ± 0.22 12.83 ± 0.96 7.58 ± 1.01 11.28 ± 0.77 38.54 ± 0.96
qlbtpy 40.23 ± 1.89 39.23 ± 1.75 45.26 ± 2.11 49.99 ± 1.33 30.58 ± 0.34
qlbtpy-Pt 12.09 ± 1.03 18.69 ± 0.49 15.29 ± 1.14 20.62 ± 1.72 35.03 ± 2.25
cis-Pt(DMSO)2Cl2 >150 >150 >150 >150 >150
cisplatinb 14.11 ± 1.56 12.18 ± 1.71 11.09 ± 1.88 17.36 ± 1.16 19.33 ± 1.14

The IC50 was defined as mean ± SD (standard deviation of the average value) from five independent assays.
198 Q.-P. Qin et al. / European Journal of Medicinal Chemistry 170 (2019) 195e202

Fig. 5. The change in cell cycle and related-proteins in NCIeH460 cancer cells after treatment with tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) for 24 h (AC) Cell cycle status of
NCIeH460 cancer cells after treatment with tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) were analyzed by flow cytometry. (D and E) Cell cycle related-proteins in tpbtpy-Pt
(0.35 mM) and dthbtpy-Pt (5.53 mM) treated cells were detected by Western blot.

potential), the increased level of intracellular Ca2þ, and increases/
decreases of the apoptosis proteins, play very important roles in
drug induced cancer cell apoptosis [68e86]. Therefore, the effects
of tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) on mitochondrial
properties of NCIeH460 cancer cells were examined by flow-
cytometry, fluorescence microscopy, and Western blot. As shown
in Fig. 6, upon treatment of NCIeH460 cells with tpbtpy-Pt
(0.35 mM) and dthbtpy-Pt (5.53 mM), the fluorescent intensity
decreased remarkably (low DJm, from right to left), as compared
with the control group (high DJm in the control group leads to JC-1
emission of red fluorescence). After treatment of NCIeH460 tumor
cells, the fluorescence of DCF (Fig. 7aec) and Fluo-3 AM (Fig. 7df)
increased. These data suggested that tpbtpy-Pt (0.35 mM) and
dthbtpy-Pt (5.53 mM) can increase intracellular free Ca2þ and ROS
levels. Furthermore, levels of bcl-2 were decreased, and levels of
cytochrome c and apaf-1 were increased after treatment of
NCIeH460 cells with tpbtpy-Pt (0.35 mM) and dthbtpy-Pt
(5.53 mM) (Fig. 8). In summary, these complexes induced
NCIeH460 cell apoptosis through mitochondrial dysfunction
pathways and tpbtpy-Pt showed stronger effect than dthbtpy-Pt.
2.6. Cell apoptosis with Annex V/PI double staining method
Based on the above results, PI/Annex V double staining was used
to evaluate tumor cell apoptosis upon treatment with tpbtpy-Pt
(0.35 mM) and dthbtpy-Pt (5.53 mM) for 24 h in NCIeH460 cells. As
shown in Fig. 9, NCIeH460 cancer cells were treated with tpbtpy-
Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) for 24 h. The percentage of
apoptotic (Q2þQ4) cells increased to 98.5% for tpbtpy-Pt, and 19.5%
for dthbtpy-Pt as compared with the control (8.7%). NCIeH460 cell
apoptosis was greater when treated with tpbtpy-Pt as compared
with treatment with dthbtpy-Pt.
2.7. The tpbtpy-Pt suppressed NCIeH460 tumor xenograft growth
in vivo
The in vivo anti-cancer activity of tpbtpy-Pt was examined.
Nude mice bearing NCIeH460 cell xenografts were treated with the
highly soluble tpbtpy-Pt at a dose of 10.0 mg/kg once every 2 days
by intraperitoneal injection over 13 days with no adverse effects
observed [47,60,85,87e95]. The treatment was found to inhibit
tumor growth by 70.1% (p < 0.05) as compared to with the vehicle
group (5% DMSO in saline, v/v), which exhibited better anticancer
activity than that of cisplatin (tumor growth inhibition (TGI), 31.8%;
tumor growth inhibition rate (IR), 25.5%) [88]. Importantly, no
significant body weight loss, and no other adverse effects and
mouse death were observed after treatment with tpbtpy-Pt
(10.0 mg/kg/q2d) at these doses (Fig. 10AeD and Table S10S12).

Fig. 6. The changes of DJm were studied after NCIeH460 tumor cells were treated with tpbtpy-Pt (0.35 mM, a) and dthbtpy-Pt (5.53 mM, b) for 24 h. (a) The cancer cells were
imaged by a flow-cytometry.
 Q.-P. Qin et al. / European Journal of Medicinal Chemistry 170 (2019) 195e202
Fig. 7. Intracellular ROS (ac) and Ca2þ (df) levels in NCIeH460 cells (a,d) exposed to tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) for 24 h. The fluorescent intensity of DCF and
Fluo-3 AM were determined by fluorescence microscopy (400  ).
Fig. 8. (A and B) Western blot analysis of bcl-2, cytochrome c, and apaf-1 in
NCIeH460 cells treated with tpbtpy-Pt (0.35 mM) and dthbtpy-Pt (5.53 mM) for 24 h.
b-actin was used as the internal control.
Fig. 9. Apoptosis in NCHeH460 cells upon (a) exposure to control conditions, (b)
tpbtpy-Pt (0.35 mM), and (c) dthbtpy-Pt (5.53 mM) for 24 h as determined by flow
cytometry.
3. Conclusion
In summary, three novel binuclear platinum(II) complexes,
[Pt(tpbtpy)Cl][Pt(DMSO)Cl3] (tpbtpy-Pt), [Pt(dthbtpy)Cl]
[Pt(DMSO)Cl3],CH3OH (dthbtpy-Pt) and [Pt(qlbtpy)Cl][Pt(DMSO)
Cl3],CH3OH (qlbtpy-Pt) with 40 -(3-thiophenecarboxaldehyde)-
2,20 :60 ,200 -terpyridine (tpbtpy), 40 -(3,5-bis(1,1- dimethylethyl)-2-
hydroxy-benzaldehyde)-2,20 :60 ,200 -terpyridine (dthbtpy) and 40 -
(2-quinolinecarboxaldehyde)-2,20 :60 ,200 -terpyridine (qlbtpy) as the
respective ligands were synthesized and fully characterized by
spectroscopic methods. MTT assay of tpbtpy-Pt, dthbtpy-Pt and
qlbtpy-Pt showed excellent IC50 values in the range of
0.35e12.09 mM, comparable to that of cisplatin (14.11 ± 1.56 mM).
Interestingly, tpbtpy-Pt and dthbtpy-Pt exerted antitumor effects,
including induction of mitochondrial dysfunction, inhibition of
telomerase activity and related-proteins, and induction of cell cycle
arrest at G1 phase in the order of tpbtpy-Pt > dthbtpy-Pt. In
199
addition, tpbtpy-Pt significantly inhibited NCIeH460 tumor
xenograft growth (TGI ¼ 70.1%, p < 0.05), suggesting that it has the
potential to be further developed as a Pt-based anticancer drug.
4. Experimental methods
4.1. Synthesis
4.1.1. Synthesis of the tpbtpy, dthbtpy and qlbtpy ligands
Synthesis of 40 -(3-thiophenecarboxaldehyde)-2,20 :60 ,200 -terpyr-
idine (tpbtpy), 40 -(3,5-bis(1,1-dimethylethyl)-2-hydroxy-benzalde-
hyde)-2,20 :60 ,200 -terpyridine (dthbtpy) and 40 -(2-
quinolinecarboxaldehyde)-2,20 :60 ,200 -terpyridine (qlbtpy) were
carried out according to published methods [42e44].
4.1.2. Synthesis and characterization of tpbtpy-Pt, dthbtpy-Pt and
qlbtpy-Pt
cis-Pt(DMSO)2Cl2 (2.0 mmol) was mixed with 1.0 mmol 40 -(3-
thiophenecarboxaldehyde)-2,20 :60 ,200 -terpyridine (tpbtpy), 40 -(3,5-
bis(1,1-dimethylethyl)-2-hydroxy-benzaldehyde)-2,20 :60 ,200 -terpyr-
idine (dthbtpy) and 40 -(2-quinolinecarboxaldehyde)-2,20 :60 ,200 -ter-
pyridine (qlbtpy) in CH3OH (25.0 mL) and DMSO (0.2 mL) at 60  C
for 6.0 h to yield red brown rod-shaped crystals of [Pt(tpbtpy)Cl]
[Pt(DMSO)Cl3] (tpbtpy-Pt), [Pt(dthbtpy)Cl][Pt(DMSO)Cl3],CH3OH
(dthbtpy-Pt) and [Pt(qlbtpy)Cl][Pt(DMSO)Cl3],CH3OH (qlbtpy-Pt),
which were isolated and characterized.
Data for tpbtpy-Pt. Yield: 90.89%. ESI-MS: m/z ¼ 545.9 for
[tpbtpy þ Pt þ Cl]þ. IR (KBr): 3427, 3078, 2999, 2912, 1610, 1559,
1523, 1476, 1439, 1371, 1243, 1130, 1023, 876, 783, 750, 716,
570 cm1. Elemental analysis: calcd (%) for C21H19Cl4OPt2S2: C
27.25, H 2.07, N 4.54; found: C 27.20, H 2.11, N 4.52.1H NMR
(500 MHz, DMSO‑d6) d 8.93 (d, J ¼ 2.1 Hz, 2H), 8.86e8.79 (m, 2H),
8.78 (d, J ¼ 7.9 Hz, 2H), 8.74 (dd, J ¼ 2.9, 1.3 Hz, 1H), 8.50 (td, J ¼ 7.8,
1.7 Hz, 2H), 8.06 (d, J ¼ 5.1 Hz, 1H), 7.90 (tt, J ¼ 5.0, 1.8 Hz, 3H), 2.54
(s, 6H).
Data for dthbtpy-Pt. Yield: 92.06%. ESI-MS: m/z ¼ 668.0 for
[dthbtpy þ Pt þ Cl]þ. IR (KBr): 3787, 3434, 2955, 1608, 1478, 1413,
1137, 1018, 785, 658, 509, 442 cm1. Elemental analysis: calcd (%)
for C31H37Cl4O2Pt2S: C 35.54, H 3.56, N 4.01; found: C 35.52, H 3.60,
N 3.98.1H NMR (500 MHz, DMSO‑d6) d 8.98 (dd, J ¼ 5.7, 1.5 Hz, 2H),
8.79e8.73 (m, 4H), 8.70 (s, 1H), 8.52 (td, J ¼ 7.9, 1.6 Hz, 2H), 7.98
(ddd, J ¼ 7.5, 5.6, 1.4 Hz, 2H), 7.44 (d, J ¼ 2.4 Hz, 1H), 7.28 (d,
J ¼ 2.3 Hz, 1H), 2.54 (s, 6H), 1.46 (s, 9H), 1.34 (s, 9H).
200 Q.-P. Qin et al. / European Journal of Medicinal Chemistry 170 (2019) 195e202
Fig. 10. The tpbtpy-Pt complex suppressed NCIeH460 tumor xenograft growth in vivo. (A) The tumor volumes of NCIeH460 xenograft-bearing mice after treatment with tpbtpy-Pt
(10.0 mg/kg/q2d) via intraperitoneal injection of six doses over 13 days. (B) Body weight of mice in the tpbtpy-Pt (10.0 mg/kg/q2d) and vehicle (5% DMSO in saline, v/v) groups. (C)
Tumor weight of tpbtpy-Pt (10.0 mg/kg/q2d) and vehicle (5% DMSO in saline, v/v) groups were recorded after 13 days. (**) P < 0.05, drug-treated group vs. the vehicle control. (D)
Tumors treated with tpbtpy-Pt (10.0 mg/kg/q2d) and vehicle (5% DMSO in saline, v/v) were dissected from the mice.
Data for qlbtpy-Pt. Yield: 85.62%. ESI-MS: m/z ¼ 590.9 for
[qlbtpy þ Pt þ Cl]þ. IR (KBr): 3493, 3056, 1606, 1551, 1477, 1416,
1142, 1123, 11032, 1018, 834, 783, 752, 690, 630, 477, 441 cm1.
Elemental analysis: calcd (%) for C26H22Cl4OPt2S: C 32.18, H 2.28, N
5.77; found: C 32.14, H 2.26, N 5.80.1H NMR (500 MHz, DMSO‑d6)
d 9.39 (d, J ¼ 13.2 Hz, 2H), 8.95 (dd, J ¼ 22.7, 6.8 Hz, 4H), 8.77 (d,
J ¼ 8.6 Hz, 1H), 8.64e8.54 (m, 3H), 8.27 (d, J ¼ 8.5 Hz, 2H), 8.14 (d,
J ¼ 8.2 Hz, 1H), 8.07e7.92 (m, 3H), 7.79 (s, 1H), 3.32 (s, 6H).
4.2. Methods
The in vitro anti-tumor activity of tpbtpy-Pt (0.35 mM) and
dthbtpy-Pt (5.53 mM) was assessed in NCIeH460 cells after 24 h
treatment as described by Ulukaya, Chao, Sadler, Che, and Liang
[47,48,55,63,83,90]. In vivo antitumor activity of tpbtpy-Pt
(10.0 mg/kg/q2d) against a NCIeH460 tumor xenograft was deter-
mined as reported by Che and Samuelson [47,60,85,87e94].
Acknowledgement
We thank the National Natural Science Foundation of China
(Nos. 21867017, 21861014 and 21761033), the Natural Science
Foundation of Guangxi (No. 2018GXNSFBA138021,
2018GXNSFBA281188 and 2016GXNSFBA380237), the Innovative
Team & Outstanding Talent Program of Colleges and Universities in
Guangxi (2014-49 and 2017-38), the basic skills improvement
project for the young and middle-aged teachers in Guangxi colleges
and universities (2019KY0605 and 2019KY1163) as well as the
scientific research project of Guilin Normal College (KYA201804)
for the financial support.
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.ejmech.2019.03.014.
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