Triphenyltin(IV) acylhydrazone compounds: Synthesis, structure and bioactivity

Article abstract of DOI:10.1016/j.jinorgbio.2018.11.011

Four new triphenyltin(IV) acylhydrazone compounds of the type Ph₃SnCH₂CH₂CONHN=R (where Ph = phenyl; R = isopropyl, isobutyl, cyclopentyl and cyclooctyl) were synthesized and characterized by elemental analysis, infrared s

Full text of DOI:10.1016/j.jinorgbio.2018.11.011

Accepted Manuscript
Triphenyltin acylhydrazone compounds: Synthesis, structure and
bioactivity
Xicheng Liu, Yanhua Tang, Xiangdong He, Xingxing Ge, Jiao
Liu, Xiaoyu Meng, Mingxiao Shao, Yingmin Jin, Laijin Tian, Zhe
Liu
PII:
S0162-0134(18)30378-7
DOI:
Reference:
https://doi.org/10.1016/j.jinorgbio.2018.11.011
JIB 10598
To appear in:
Journal of Inorganic Biochemistry
Received date:
Revised date:
Accepted date:
27 June 2018
17 November 2018
17 November 2018
Please cite this article as: Xicheng Liu, Yanhua Tang, Xiangdong He, Xingxing Ge, Jiao
Liu, Xiaoyu Meng, Mingxiao Shao, Yingmin Jin, Laijin Tian, Zhe Liu , Triphenyltin
acylhydrazone compounds: Synthesis, structure and bioactivity. Jib (2018), https://doi.org/
10.1016/j.jinorgbio.2018.11.011
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ACCEPTED MANUSCRIPT
Triphenyltin Acylhydrazone Compounds: Synthesis, Structure and
Bioactivity
Xicheng Liu+a*, Yanhua Tang+a, Xiangdong Hea, Xingxing Gea, Jiao Liua, Xiaoyu Menga, Mingxiao Shaoa,
Yingmin Jina, Laijin Tiana*, Zhe Liua*
a
Institute of Anticancer Agents Development and Theranostic Application, The Key Laboratory of Life-Organic
Analysis and Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, Department of
Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, China.
Corresponding authors: E-mail: chemlxc@163.com (Xicheng Liu);tianlaijin@163.com (Laijin Tian);
liuzheqd@163.com (Zhe Liu).
+ X. C. Liu and Y. H. Tang have equivalent contribution.
Abstract: Four new triphenyltin acylhydrazone compounds of the type Ph3SnCH2CH2CONHN=R
(where Ph = phenyl; R = isopropyl, isobutyl, cyclopentyl and cyclooctyl) were synthesized and
characterized by elemental analysis, infrared spectrum (IR), nuclear magnetic resonance spectrum
(NMR) and mass spectrum (MS). The crystal structures were determined and showed that tin
atoms were four-coordinated and adopted a pseudo-tetrahedron configuration. Tin compounds
show excellent BSA binding properties, and can oxidize NADH to generate reactive oxygen
species (ROS), which inducing apoptosis effectively. Bioassay results indicated that tin
compounds have stronger cytotoxic activity against A549 human lung cancer cells compared with
cis-platin used clinically, and showing some selectivity.
Key words: Triphenyltin; Acylhydrazone; Antitumor
Since Rosenberg et al. found cis-platin has antitumor activity in 1969 [1], the medicinal
properties of metal complexes have attracted wide attention and opened up a new field of research.
Now, platinum complexes have widely used to treat various cancer. [2] The treatment with
platinum-based drugs or platinum-related drugs accounted for more than 50% among all
chemotherapy regiments. However widely used it is today, the platinum antitumor drugs also
show some of the increasingly prominent problem-severe side effects [3], which urge researchers
to seek the alternative antitumor drugs with more efficient, better targeted, and the antitumor
mechanism different from platinum drugs. [4] Among these, organotin compound is one of the
most active metal organic compound in the last few decades. A large number of organotin
compounds have shown higher anttumor activity than the widely used cis-platin antitumor drugs
by experiments in vitro. [5-10]
Schiff base mainly refers to series of organic compounds containing the imide or methylamine
group (-RC=N-), which were usually condensed by amine and active carbonyl including the type
of amino acids, thiosemicarbazones, shrinkage amine, heterocyclic, hydrazone etc. [11] Schiff
base has the unique medicinal effect in antibacterial, bactericidal, antitumor and anti-virus. [12]
And also, Schiff base has good coordination ability with transition metal using nitrogen and
oxygen atoms inside the group. [13] More importantly, because of the special structure, Schiff
base is close to the real situation of biological system and suitable for the simulation study of life

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system, which have attracted more and more attention in recent years. [14] Acylhydrazone
derivatives are a special class of Schiff base compounds, which are formed by the condensation of
hydrazine and aldehydes or ketones. The structure is characterized by the presence of -CO-NH-
N=C- fragment in the molecule. The p-π conjugation existed in the fragment of imino group, the
carbonyl group and the amino group enhance the stability of compounds. The bigger conjugate
system increase electron liquidity of intramolecular and give such compounds special activity. [15]
Organotin compounds have strong biological toxicity, which was in relation to their organotin
parts and the types of ligands. The low molecular weight organotin can seriously damage the
immune system. The more numbers of alkyl substituents, the stronger of the hydrophobic, and the
greater of compound's toxicity. [16] However, recent research has shown that the toxicity will
decrease when the numbers of alkyl substituents increased to a certain extent, or increasing the
carbon chain length connected with tin atom and the volume of compounds. [17] Given these, in
this article, acylhydrazone units condensed by alkyl ketones and hydrazines were introduced into
the alkyl substituents to increase the carbon chain length and the volume of tin compounds (Fig.
1), which enhancing the lipid solubility, reducing the toxicity and improving antitumor activity of
tin compounds. The antitumor activities of TPT-1~TPT-4 were evaluated against A549 human
lung cancer cells by MTT assay, and cis-platin was used as the positive and negative control for
the cytotoxicity study in vitro. The anti-cancer mechanism was further studied by BSA interaction,
reaction with NADH, apoptosis assay, cell cycle analysis and ROS production.
Fig. 1 The structure of as-synthesized tin compounds TPT-1~TPT-4.
Results and Discussion
Tin compounds were obtained with cheap starting materials and a simple synthesis process.
The prepared tin compounds and synthetic routes are summarized in Fig. 1 and Scheme 1,
respectively. As-synthesized tin compounds were fully characterized by elemental analysis, IR, 1H

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NMR, MS and X-ray single crystal diffraction (ESI Fig. S1-S3). All the analytical data are
consistent with the proposed structures.
Scheme 1 Preparation of the tin compounds.
Crystal Structures
Single crystals of TPT-1, TPT-3 and TPT-4 suitable for X-ray diffraction analysis were
obtained by slow diffusion of hexane into a saturated dichloromethane solution of the compounds.
The three X-ray crystal structures are shown in Fig. 2, the crystallographic data and the selected
bond lengths, bond angles are listed in Table 1 and Table 2, respectively. Except for TPT-4, one
crystal unit cell contains two molecules for TPT-1 and TPT-3. In all of these compounds, the
coordination geometry aroud tins are described as pseudo-tetrahedron with the bond angels of C-
Sn-C changed from 101.8(5)° to 119.3(5)°. Because of the large hindrance of triphenyltin, the O(1)
of the acylhydrazone group did not coordinate with Sn(1), which are reflected in the long
separation distance of Sn(1)-ꢀO(1) (2.913(4), 2.855(3) and 3.412(5) Å for TPT-1, TPT-3 and
TPT-4, respectively, which is longer than the usual Sn-O coordination bond (2.3-2.6 Å)). [18] The
result is consistent with the analysis of the elastic vibration frequency of the carbonyl group in IR
(>1650 cm-1). [19,20] It is worth mentioning that the distance of Sn(1)-ꢀO(1) are shorter than the
van der Waals radius (3.78 Å) [21] for all tin compounds, showing that some interaction still exist
between Sn(1) and O(1), which play a certain role in the distortion of tetrahedral configuration.
The intramolecular hydrogen bonds between O(1) and hydrogen atom of N(1) existed in all
compounds. Among these, TPT-4 form dimer (ESI Fig. S4b) using intermolecular hydrogen
bonds, while TPT-1 and TPT-3 form one-dimensional (1D) chain structures (Fig. 3 and ESI Fig.
S4a). [22]

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Fig. 2 The molecular structure of TPT-1 (a), TPT-3 (b) and TPT-4 (c). The hydrogen atoms have been omitted
for clarity.
Fig. 3 The 1D chain structure of TPT-3, formed through intermolecular hydrogen bonds. All the hydrogen atoms
are omitted for clarity except for forming intermolecular hydrogen bonds.
Table 1 Crystal data and structure refinement for tin compounds.
TPT-1
TPT-3
TPT-4
C29 H34N2OSn
545.27
Empirical formula
Formula weight
Temperature (K)
C24H26N2OSn
477.16
C26H28N2OSn
503.19
295(2)
295(2)
295(2)

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Triclinic
P-1
Crystal system
Space group
a (Å)
Monoclinic
P 21/c
Monoclinic
P 21/c
7.266(2)
13.496(5)
14.437(5)
1372.1(8)
2
10.970(3)
15.136(8)
30.02(3)
4693(5)
8
11.3898(10)
14.6174(12)
31.5542(19)
4917.4(7)
8
b (Å)
c (Å)
volume (Å3)
Z
Dc (g cm-3)
μ (mm-1)
F (000)
1.351
1.359
1.320
0.953
1.104
1.057
560
1936
2048
1.42 to 26.00
0.40×0.30×0.08
1.44 to 25.25
1.38 to 26.00
 range ()
crystal size (mm)
Tot. reflections
0.30×0.12×0.04
32601
0.26×0.22×0.07
37919
10735
Uniq. reflections
GOF on F2
8461 [Rint = 0.2484]
0.842
9639 [Rint = 0.1013]
0.984
5328 [Rint = 0.0532]
0.998
R=0.0572, wR=0.1127
R=0.0748, wR=0.1547 R=0.0604, wR=0.1137
R indices [I>2 (I)]
R indices (all data)
△ρmin, △ρmax (eÅ-3)
R=0.2145, wR=0.2022 R=0.1257, wR=0.1340 R=0.0952, wR=0.1265
-0.642, 0.751
-0.409, 0.578
-0.513, 0.681
Table 2 Selected bond lengths (Å) and angles (deg.) for tin compounds.
TPT-1
TPT-3
TPT-4
Sn(1)-C(1)
2.134(14)
2.136(13)
2.145(13)
2.144(11)
2.913(4)
101.8(5)
119.3(5)
116.1(5)
105.3(6)
106.6(5)
106.3(4)
2.161(6)
2.128(6)
2.129(7)
2.142(6)
2.855(3)
103.8(2)
103.7(2)
105.0(2)
112.3(3)
113.3(3)
117.0(3)
2.146(5)
2.138(5)
2.123(6)
2.141(5)
3.412(5)
107.4(2)
110.5(2)
104.2(2)
109.2(2)
113.8(2)
111.6(2)
Sn(1)-C(7)
Sn(1)-C(13)
Sn(1)-C(19)
Sn(1)···O(1)
C(1)-Sn(1)-C(7)
C(1)-Sn(1)-C(13)
C(1)-Sn(1)-C(19)
C(7)-Sn(1)-C(13)
C(7)-Sn(1)-C(19)
C(13)-Sn(1)-C(19)
In vitro cytotoxicity

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The antitumor activities of compound 2 (raw material, triphenyltin compound substitued by
hydrazine carbonyl), TPT-1~TPT-4 were evaluated against A549 human lung cancer cells by
MTT assay, and cis-platin was used as the positive and negative control for the cytotoxicity study
in vitro under the same condition. The IC50 values (concentration at which 50% of the cell growth
is inhibited) of exposured to tin compounds after 24 h were shown in Table 3. Compared with
compound 2, target tin compounds (TPT-1~TPT-4) exhibit the higher antitumor activity. TPT-1
and TPT-4 displayed the higher activities in vitro, which were more active than the clinically
widely used cis-platin. The activity of TPT-4 is the best, indicating the terminal substituent, large
circle alkyl group seems to have a favorable effect on antitumor activity. Thus, further structure
modification based on the acylhydrazone may modulate the cytotoxicity for these compounds.
The antiproliferative activities of TPT-2 and TPT-4 were further evaluated against two
human bronchial epithelial normal cells (16HBE and BEAS-2B). As shown in Table 3, TPT-2
and TPT-4 show some selectivity for cancer cells versus normal cells, but it is not obvious.
Therefore, more structural modification is necessary to improve the selectivity in future work.
Table 3 IC50 values of Complexes 2, TPT-1~TPT-4 and cis-platin against A549 Cancer Cells and human
bronchial epithelial normal cells after 24 h exposure.
IC50 (µM)
A549
IC50 (µM)
16HBE
--
IC50 (µM)
BEAS-2B
--
Compounds
>100
2
16.9±0.1
35.0±1.4
21.1±0.8
9.2±0.1
--
--
TPT-1
64.3±4.6
49.4±0.8
TPT-2
TPT-3
--
--
35.7±3.5
16.5±0.9
TPT-4
21.3±1.7
cis-platin
--
--
Study of BSA interactions
It is necessary to understand what happened in cells by investigating the interaction between
antitumor agents and protein. [23] In blood plasma, serum albumin (SA) act the important role in
drug transport and metabolism. [24] In this work, bovine serum albumin (BSA) was used to
investigate the interaction with as-synthesized tin compounds for its structural similarity to human
serum albumin (HSA), and also easily obtained.
The ultraviolet-visible (UV-Vis) absorption spectra of BSA in the absence and presence of
tin compounds are shown in Fig. 4a (TPT-1) and ESI Fig. S5 (TPT-2~TPT-4). As shown, the
absorption peak decreased significantly and shifted toward longer wavelength at 218 nm along
with increasing the concentration of tin compounds, which was attributed to the induced
perturbation of BSA because of tin compounds. [25,26] Obvious red-shift was found at 218 nm

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attribute to the effect of the polar solvent (water). [27] The binding of BSA to complexes was also
reflected by the quenching at 278 nm, but without any shift, which indicate that tin compounds
were mainly changing the microenvironment of BSA. [28,29]
Fig. 4 (a) UV-Vis spectrum of BSA in 5 mM Tris–HCl/10 mM NaCl buffer solution (pH = 7.2) upon addition of
TPT-1 (0–10.0 μM). The arrows show the direction of changes in absorbance upon increasing the concentration of
TPT-1. Inset: wavelength from 255 to 300 nm. (b) Fluorescence spectra of BSA (0.5 μM; λex = 280 nm; λem = 343
nm) in the absence and presence of the TPT-1 (0-10.0 μM).
Fluorescence quenching is another effective method to study the interaction between
compounds and proteins. As shown in Fig. 4b (ESI Fig. S6), with the increase of TPT-1, the
fluorescence intensity of BSA decreases continuously at 343 nm. [30] To get a possible quenching
mechanism, emission quenching data have been analyzed by the classical Stern–Volmer equation
and Scatchard equation [31,32], and the important fitted data were listed in Table 4 (ESI Fig. S7
and S8). As shown, the quenching rate constant Kq for all complexes range from 2.74 × 1012 to
7.58 × 1012 M-1 s-1, which are about two orders of magnitude higher than that of a purely dynamic
quenching mechanism (2.0×1010 M-1 s-1), and indicating that a static quenching mechanism
dominates in the interaction of tin compounds and BSA. [33] The binding site number n and
binding constant Kb of all tin compounds were almost the same. But TPT-4 show the bigger Kb
(5.52×104 M-1) compared with other tin compounds, which is consistent with the conclusion that
TPT-4 has the best antitumor activity.
Table 4 The values of Kq, Kb and n for as-synthesized tin compounds.
Kb
Kq
Compounds
n
（104 M-1）
(1012 M-1S-1)
2.74
4.20
4.37
1.32
1.04
0.90
TPT-1
TPT-2
5.74
7.58
3.68
5.52
0.96
0.98
TPT-3
TPT-4
Reaction with NADH

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In many biological processes, reduced form of nicotinamide-adenine dinucleotid (NADH)
play a major role. [34] Transition metal complex can oxidize NADH to NAD+ and generate
reactive oxygen species (ROS), which is the important oxidation antitumor mechanism. [35] The
interaction between tin compounds and NADH can be detected at 259 nm and 340 nm by UV-Vis
spectrum (Fig. 5a and ESI Fig. S9). The absorption bands in 259 nm region can be assigned to the
absorption of NAD+ (produced by tin compounds capture hydrogen ions on NADH), while peaks
at 340 nm are the absorption of NADH. [36] The turnover numbers (TONs) of TPT-1 (7.18),
TPT-2 (2.15), TPT-3 (6.17) and TPT-4 (7.95) were calculated by measuring the difference at 340
nm (Fig. 5b,). TONs show that all complexes can oxidize NADH to NAD+. Among these, TPT-4
shows the best activity, which are in accord with the results obtained in vitro cytotoxicity, and also
demonstrating that the introduction of large cyclic alkanes is helpful to improve the catalytic
performance of tin compound.
Fig. 5 (a) UV-Vis spectra of the reaction of NADH (100 µM) with TPT-1 (1 µM) in 10% MeOH/90% H2O (V:V)
at 298 K for 8 h. The arrows show changes in absorbance spectra over time; (b) The TONs of as-synthesized tin
compounds.
Apoptosis assay
Compounds TPT-2 and TPT-4 are selected for more research on action mechanism. In order
to evaluate whether the cell death caused by tin complex is based on the mechanism of apoptosis,
the annexin V assay was performed. A549 lung cancer cells were treated with TPT-2 and TPT-4
at 1, 2 and 3 × IC50 for 24 h, followed by staining with annexin V/propidium iodide (PI), and
analyzed by flow cytometry. As shown in Fig. 6, cells in apoptosis stage increased with the
concentrations of tin compounds. TPT-2 and TPT-4 dramatically induce apoptosis during 24 h at
a concentration of 3 × IC50, with 36.6% and 47.5% cells undergoing apoptosis (early apoptosis +
late apoptosis) , respectively, compared with ~7% of control (ESI Table S1 and S2).

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Fig. 6 (a) Apoptosis induction detected by annexin V-FITC and PI staining and flow cytometry analysis in A549
cells after 24 h of exposure to TPT-2 and TPT-4 at various concentrations at 310 K. (b) Histogram showing
populations for A549 cells in four stages treated by TPT-2 and TPT-4. Data are quoted as mean ± SD of three
replicates.
Cell Cycle Analysis
Cell cycle arrest analysis for TPT-2 and TPT-4 against A549 lung cancer cells were
investigated by flow cytometry. As shown in Fig. 7 (ESI Table S3 and S4), the cell cycle
progression was analyzed at 0.25, 0.5 and 1.0 equipotent concentrations of IC50 for 24 h. At a
concentration of 1.0 × IC50, compared with control, the percentages of cells in the S phase and G1
phase increased from 25.0% and 10.2% to 43.9% and 21.1% when exposed to TPT-2. For TPT-4,
at a concentration of 1.0 × IC50, the percentages of cells in the S phase of the cell cycle increased
from 22.6% to 36.1%. Compared with the untreated control, complex TPT-4 disturbs the cell
cycle at S and G1 phase, while TPT-2 mainly disturbing the cell cycle at S phase. The results
indicate that these tin compounds can effectively disturb the cell growth cycle progression, and
achieving the purpose of apoptosis.

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Fig. 7 (a) Cell cycle analysis of A549 lung cancer cells after 24 h of exposure to TPT-2 and TPT-4 at the
indicated concentrations by flow cytometry using PI staining. (b) Populations in each cell cycle phase for control,
TPT-2 and TPT-4.
Induction of ROS
Excessive production of reactive oxygen species (ROS) often led to oxidative stress and
damages to cells. [34] The levels of ROS induced by TPT-2 and TPT-4 were investigated by flow
cytometry analysis (Fig.8, ESI Table S5 and S6). A marked increase of ROS levels in A549 cells
were observed after 24 h exposed to TPT-2 and TPT-4 even at a concentration of 0.25 × IC50,
around 75% of A549 cells were in high ROS levels. These observations are consistent with the
proposed mechanism of action for TPT-2 and TPT-4, which is based on the disruption of the
cellular redox balance. [37] The significant increase of ROS levels may provide a basis for killing
cancer cells.

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Fig. 8 ROS induction in A549 cancer cells caused by TPT-2 and TPT-4 at concentration of 0.25 and 0.50 × IC50.
(a) FL2 histogram for negative control (cells untreated), positive control, TPT-2 and TPT-4. (b) Populations of
cells in low and high level of ROS after treatment of TPT-2 and TPT-4. Data are quoted as mean ± SD of three
replicates.
Conclusion
In this study, four triphenyltin acylhydrazone compounds were synthesized with cheap
starting materials and simple synthesis processes. The introduction of acylhydrazone group to the
alkyl substituent increase the carbon chain length and the volume, meanwhile enhancing the lipid
solubility and improving antitumor activity of tin compounds. TPT-4 showed the best activity
towards A549 lung cancer cells (IC50: 9.2 µM) and some selectivity. The UV-Vis and PL test
show all compounds can effectively bind with BSA, which indicate tin compounds can transport
through blood plasma. Tin compounds can oxidize NADH to generate ROS and lead to apoptosis.
The results of flow cytometry indicate that the compounds can disturb the cell growth cycle,
which further confirmed the apoptosis mechanism. Above all, triphenyltin acylhydrazone
compounds could be a promising candidate for further evaluation as antitumor drugs.
Experimental Section
General
All chemicals were commercial grade and used without purifcation. Triphenyltin chloride,
Lithium Aluminum Hydride, methyl acrylate, hydrazine hydrate (80%), acetone, butanone,
cyclopentanone, Cyclooctanone and other organic reagents were purchased from Shanghai
Pengteng Fine Chemical Co., Ltd. For the biological experiments, DMEM medium, fetal bovine
serum, reduced form of nicotinamide-adenine dinucleotid (NADH) were purchased from Sangon
Biotech. A549 lung cancer cells were obtained from Shanghai Institute of Biochemistry and Cell

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Biology (SIBCB). Carbon, hydrogen, oxygen and nitrogen analyses were performed using a
Perkin Elmer 2400 Series II elemental analyzer (Perkin Elmer, Waltham, MA, USA). IR spectra
were recorded on a Nicolet Nexus 470 FT-IR spectrophotometer using KBr discs in therange
1
4000-400 cm-1 (Thermo Nicolet Corporation, Madison, WI, USA). H NMR spectral data were
collected using a Bruker Avance DPX 500 NMR spectrometer (Bruker Corporation, Switzerland)
with CDCl3 as the solvent and tetramethylsilane (TMS) as the internal standard. Intensity data for
the crystals were measured at 295(2) K on a Bruker Smart Apex area-detector fitted with graphite
monochromatized Mo-K radiation (0.71073 Å) using the  and  scan technique. UV-Vis
spectroscopy was performed on a TU-1901 UV spectrometer. The fluorescence spectra were
collected by a fluorescence spectrophotometer (F-4600, Hitachi) with a 400 V voltage and 5 nm
slit width for both excitation and emission. Apoptosis and ROS determination were carried out
using the Annexin V-FITC Apoptosis Detection Kit (Beyotime Institute of Biotechnology, China).
MTT assay were measured using a microplate reader (DNM-9606, Perlong Medical, Beijing,
China) at an absorbance of 570 nm.
Synthesis
Synthesis of β-methoxycarbonyl ethyl triphenyltin (1)
1
Compound 1 was prepared according to literature methods. [38,39] H NMR (500 MHz,
CDCl3)/: 7.54-7.56 (m, 6H), 7.36-7.38 (m, 9H), 3.47 (s, 3H), 2.68 (t, J = 7.8 Hz, 2H), 1.67 (t, J =
7.8 Hz, 2H).
Synthesis of β-hydrazinocarbonyl ethyl triphenyltin (2)
Compound 1 (2.0 g, 4.5 mmol) and methanol (30 mL) were added into a 250 mL three-
necked bottle under N2, followed by stirring and heating to ebullience for 20 min. Hydrazine
hydrate (50%, 30 mL) was added dropwise to above solution at 80 °C. The reaction stopped until
compound 1 was consumed completely (monitored by thin-layer chromatography). Crude product
was isolated by filtration, and recrystallized from 60 mL mixture of chloroform and petroleum
ether (V/V=1:1) to obtain a acicular colorless crystal. Yield: 80.1%; Mp, 144.2-144.5 °C; IR
(KBr): 1680 cm-1 (C=O); 1H NMR (500 MHz, CDCl3) δ 7.56 (dt, J = 5.7, 2.6 Hz, 6H), 7.38 – 7.35
(m, 9H), 2.50 (t, J = 7.8 Hz, 2H), 1.67 (t, J = 7.8 Hz, 2H); ESI-MS (m/z): [M+Na]+ Calcd for
C21H22N2OSn, 461.08; Found 461.25.
Synthesis of β-acylhydrazone ethyl triphenyltin (TPT)
To a solvent of trichloromethane, Compound 2 (1 eq) and corresponding ketone (1 eq.) were
added. The mixture was stirred at 80 °C until compound 2 was consumed completely (monitored
by thin-layer chromatography). After rotary evaporation, the crude product was recrystallized
from the mixture of chloroform and petroleum ether (V/V=1:1). The characterization data were
described for each case as follows:

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Ph3SnCH2CH2CONHN=C(CH3)2 (TPT-1): Colorless crystal; Yield: 87.3%; Mp,134.9-136.5 °C;
1
IR (KBr): 1663 cm-1 (C=O) 1541 cm-1 (C=N); H NMR (500 MHz, CDCl3) δ7.97 (s, 1H), 7.61 –
7.56 (m, 6H), 7.34 (dd, J = 4.8, 1.8 Hz, 9H), 3.03 (t, J = 7.7 Hz, 2H), 2.31 (t, J = 7.3 Hz, 2H), 1.69
(d, J = 5.7 Hz, 6H). Elemental analysis: Found: C 61.09, H 5.78, N 5.65, O 3.30%, anal. Calcd. for
C25H28N2OSn: C 61.13, H 5.75, N 5.70, O 3.26%. ESI-MS (m/z): [M+Na]+ Calcd for
C24H26N2OSn, 501.11; Found 501.33.
Ph3SnCH2CH2CONHN=C(CH3)(C2H5) (TPT-2): Colorless crystal; Yield: 80.6% ; Mp, 129.3-
130.5 °C; IR (KBr): 1660 cm-1 (C=O), 1548 cm-1 (C=N); 1H NMR (500 MHz, CDCl3) δ 8.04 (d, J
= 21.6 Hz, 1H), 7.59 (dd, J = 5.3, 1.9 Hz, 6H), 7.34 (dd, J = 8.1, 5.0 Hz, 9H), 3.04 (t, J = 7.8 Hz,
2H), 2.22 (q, J = 7.4 Hz, 2H), 1.72 – 1.66 (m, 5H), 1.04 (t, J = 7.2 Hz, 3H). Elemental analysis:
Found: C 61.18, H 5.69, N 5.68, O 3.25%, anal. calcd. for C25H28N2OSn: C 61.13, H 5.75, N 5.70,
O 3.26%. ESI-MS (m/z): [M+Na]+ Calcd for C25H28N2OSn, 515.12; Found 515.33.
Ph3SnCH2CH2CONHN=CC4H8 (TPT-3): Colorless crystal; Yield: 81.6%; Mp, 146.3-147.6 °C;
1
IR (KBr): 1652 cm-1 (C=O), 1548 cm-1 (C=N); H NMR (500 MHz, CDCl3) δ 7.77 (s, 1H), 7.59
(dd, J = 6.3, 3.0 Hz, 6H), 7.34 (dd, J = 4.8, 1.8 Hz, 9H), 3.02 (t, J = 7.6 Hz, 2H), 2.33 (t, J = 7.2
Hz, 2H), 2.03 (t, J = 6.9 Hz, 2H), 1.85 – 1.78 (m, 3H), 1.73 (dt, J = 10.0, 4.9 Hz, 3H). Elemental
analysis: Found: C 62.11, H 5.59, N 5.61, O 3.21%, anal.calcd for C26H28N2OSn: C 62.06, H 5.61,
N 5.57, O 3.18%. ESI-MS (m/z): [M+Na]+ Calcd for C26H28N2OSn, 527.12; Found 527.33.
Ph3SnCH2CH2CONHN=CC7H12 (TPT-4): Colorless crystal; Yield: 78.2%; Mp, 94.3-94.7 °C;
IR (KBr): 1666 cm-1 (C=O), 1428 cm-1 (C=N); 1H NMR (500 MHz, CDCl3) δ 8.16 (s, 1H), 7.59
(dd, J = 6.3, 3.1 Hz, 6H), 7.33 (td, J = 4.8, 2.0 Hz, 9H), 3.03 (t, J = 7.7 Hz, 2H), 2.32 – 2.26 (m,
2H), 2.20 – 2.14 (m, 2H), 1.61 (dt, J = 15.5, 6.0 Hz, 4H), 1.55 – 1.42 (m, 6H), 1.35 (dd, J = 14.0,
8.9 Hz, 2H); Elemental analysis: Found: C 63.91, H 6.26, N 5.12, O 2.92%, anal.calcd for
C29H34N2OSn: C 63.87, H 6.28, N 5.14, O 2.93%. ESI-MS (m/z): [M+Na]+ Calcd for
C29H34N2OSn, 569.17; Found 569.42.
Acknowledgments
We thank Student's Platform for Innovation and Entrepreneurship Training Program of Shandong
Province (201710446042), the National Natural Science Foundation of China (Grant No.
21671118) and the Taishan Scholars Program for support.
Notes
The authors declare no competing financial interest.
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Graphical abstract
A serie of triphenyltin acylhydrazone compounds have been synthesized
and characterized. All compounds showed potent cytotoxicity, excellent
bovine serum albumin binding properties, and oxidizing NADH to NAD+
efficiently. Compounds can induce apoptosis effectively and led to the
emergence of reactive oxygen species in cells.

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Highlights
 Triphenyltin acylhydrazone compounds show potential antitumor activity towards
A549 cells.
 Triphenyltin acylhydrazone compounds can disturb the cell growth cycle and
induce apoptosis.