Synthesis and evaluation of selenium-containing indole chalcone and diarylketone derivatives as tubulin polymerization inhibition agents

Article abstract of DOI:10.1039/c7ob01655g

Sixteen new selenium-containing indole chalcone and diarylketone derivatives were synthesized and evaluated as tubulin polymerization inhibitors. Among them, compound 25b exhibited the most potent antiproliferative activities against six human cancer cell lines with IC₅₀ values of 0.004-0.022 μM. A microtubule dynamics assay and an immunofluorescence assay confirmed that 25b could effectively inhibit tubulin polymerization (IC₅₀ = 2.1 ± 0.27 μM). Further cellular mechanism studies revealed that 25b induced G2/M phase arrest, which was further evidenced by the decrease in the mitochondrial membrane potential (MMP).

Full text of DOI:10.1039/c7ob01655g

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Takeharu Haino et al.
Solvent-induced emission of organogels based on tris(phenylisoxazolyl)
benzene
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Synthesis and Evaluation of Selenium-containing Indole Chalcone
and Diarylketone Derivatives as Tubulin Polymerization Inhibition
Agents
Received 00th January 20xx,
Accepted 00th January 20xx
Shun zhang, Baijiao An, Jiayan Li, Jinhui Hu, Ling Huang,* Xingshu Li,* and Albert S. C. Chan
DOI: 10.1039/x0xx00000x
Sixteen new selenium-containing indole chalcone and diarylketone derivatives were synthesized and evaluated as tubulin
polymerization inhibitor. Among them, compound 25b, exhibited most potent antiproliferative activities against six human
cancer cell lines with IC50 values of 0.004 - 0.022μΜ. Microtubule dynamics assay and immunofluorescence assay
confirmed that 25b could effectively inhibit tubulin polymerization (IC50 = 2.1± 0.27 μM). Further Cellular mechanism
studies revealed that 25b induced G2/M phase arrest, which was further evidenced by the decrease in the mitochondrial
membrane potential (MMP).
www.rsc.org/
experiments have indicated that organoselenium has a higher
bio-disposability and biological activity than Se in inorganic
compounds. In recent years, the potential cancer therapeutic
Introduction
Cancer is a serious disease that is the second leading cause of
human death after cardiovascular disorders. In past decades,
many anticancer agents targeting different enzymes or
receptors have been developed or are currently being
developed. Among them, microtubule stabilizing or
microtubule destabilizing agents have attracted much
attention because microtubules play vital roles in the
regulation of mitosis, disruption of microtubule dynamics,
induction of cell cycle arrest in G2/M phase, formation of
abnormal mitotic spindles, and signals for apoptosis.1-2
Colchicine3 is a typical tubulin inhibitor targeting the colchicine
binding site, and Combretastatin A-4 (CA-4) (Fig. 1), which was
first isolated from the bark of the African willow tree
Combretum caffrum in 1989, exhibits potent tubulin
polymerization inhibition and binds to the same binding site as
colchicine.4,5 To develop novel and more effective anticancer
agents, many studies on the modification of CA-4 have
appeared in recent years.6-10
effects of selenium is attracting more and more attention, and
several organoselenium compounds have been shown to
inhibit cancer cell growth in various xenograft rodent models
for different cancer types. 13-17 Also, some reports showed that
selenium has a protective effect in chemotherapy of some
types of cancer.
Indole and its derivatives have a wide range of biological
activities. Some naturally occurring indole alkaloids including
vincristine, vinblastine, vinorelbine and vindesine have gained
FDA approval as drugs for anti-tumor activity. Synthetic drugs
such as indomethacin (NSAID), ondansetron (chemotherapy
induced
nausea
and
vomiting),
fluvastatin
(hypercholesterolemia), and zafirlukast (leukotriene receptor
antagonist), have also reached the patient's bedside.18
Recently, indole derivatives were also studied as tubulin
inhibitor. Jang-Yang Chang et al reported 4- and 5-aroylindoles
(Fig. 1, compound
work, we disclosed the evaluation of a series of indole-
chalcone derivatives (Fig. 1, compound ) exerting effective
A
) as antitubulin agents.19 In our previous
Selenium (Se) is an essential trace mineral nutrient with
multiple roles in the growth and function of living animal cells,
e.g., providing protection from free radical-induced cell
damage.11-12 It is known that there are twenty-five
selenoproteins in the human body with specific biological
functions. Selenium compounds have been used as promising
chemo-preventive agents for some diseases. Some animal
B
antitumor activity through microtubule destabilization.20
Inspired by the extensive pharmacological activities of indole
analogues and biological activities of Selenium, herein, we
report the synthesis of a series of selenium-containing indole
chalcone derivatives (Fig 1) and diarylketone derivatives (Fig
1). Subsequently, their antitumor activities are evaluated in
vitro, and their underlying cytotoxic mechanisms are also
elucidated.
School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006,
China;
*For L. Huang: Tel.: +086-20-3994-3051; Fax: +086-20-3994-3051; e-mail:
huangl72@mail.sysu.edu.cn; For X. Sh. Li: Fax: +086-20-3994-3050; Tel.: +086-20-
3994-3050; E-mail: lixsh@mail.sysu.edu.cn
Electronic Supplementary Information (ESI) available: [details of any
supplementary information available should be included here]. See
DOI: 10.1039/x0xx00000x
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Fig 1 Structure of CA-4, representative compound of aroylindoles (A) and indole-
chalcone derivatives (B), structural general formula of selenium-containing indole
chalcone derivatives and diarylketone derivatives.
Results and discussion
Chemistry
The synthetic routes of the target compounds including twelve
chalcone derivatives and four diarylketone derivatives are
depicted in schemes 1-5. First, the selenium-containing key
Scheme 1. The synthesis of intermediate compound 6. Reagents and conditions: (a)
HNO3, AcOH, 3 h, r.t. (b) Dimethylsulfate, K2CO3, acetone, 2d, 80°C. (c) Fe, AcOH, 8 h,
r.t. (d) i: HCl, H2O, NaNO2, 30 min, -5°C, ii: NaOAc, 30 min, 0°C, iii: KSeCN, 3 h, 0°C-r.t.
(e) NaBH4, CH3I, EtOH, 2 min, r.t.
intermediate
hydroxy-3-methoxyacetophenone
nitric acid in the presence of acetic acid, compound
6 was prepared from commercially available 4-
1. By the reaction of
1
2
with
was
obtained in good yield, which was methylated by
dimethylsulfate to afford . The hydrogenation of 3 with iron
in the presence of acetic acid gave amine intermediate
which was diazotized and reacted immediately with KSeCN to
provide ketone intermediate . The reaction of with CH3I and
NaBH4 afforded quickly and in good yield. Indole-3-
3
4,
5
5
6
carbaldehyde derivatives 8a-8c, 10a-10c and 11a-11c were
prepared as shown in scheme 2. Commercially available
compounds
to give . The reaction of commercially available substituted
indole with POCl3 and DMF afforded 10, which were treated
with methyl iodide to provide the N-methyl products 11a-11c
(scheme 2) Finally, the aldol condensation of selenium-
containing intermediate with indole-3-carbaldehyde
derivatives 10 or 11 gave target compounds 12 13 and 14
(scheme 3).
7 reacted with methyl iodide and sodium hydride
8
9
.
6
7
,
8,
,
The syntheses of diarylketone derivatives are summarized in
schemes 4-5. First, selenium-containing bromobenzene
derivatives 21 were prepared with the same procedures as the
synthesis of
6 using 2-methoxy phenol as the starting material
(scheme 4). Then, by treating with phenylsulfonyl chloride,
Scheme 2. The synthesis of intermediate compounds 8, 10 and 11. Reagents and
indole-3-carbaldehyde analogues 7a-7c were converted to
conditions: (f) CH3I, NaH, THF, 12 h, r.t. (g) POCl3, DMF, 8 h, r.t.
their N-phenylsulfonyl derivatives 22a-22c. Finally, target
compounds 25a-25c and 27 were obtained by treating
intermediate 21 with 22a-22c or 8b in the presence of n-BuLi
at -78°C, oxidizing the resulting alcohol by IBX and removing
the protecting group from 23a-23c (scheme 5).
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Scheme 5. The synthesis of target compounds 25a-25c and 27. Reagents and
conditions: (j) phenylsulfonyl chloride, NaH, THF, 10 h, 0°C. (k) 1-(phenylsulfonyl)-1H-
indole-4-carbaldehyde or 1-methyl-1H-indole-4-carbaldehyde, n-BuLi, dry THF, -78°C,
12 h. (l) IBX, THF, 2d. (m) NaOH, CH3OH, 3 h, r.t.
Scheme 3. The synthesis of target compounds 12, 13 and 14. Reagents and conditions:
(h) Piperidine, AcOH, EtOH, 2-3d, 95°C.
Biological evaluation
In vitro growth inhibition of human cancer cell lines
The antiproliferative activities of the two series of compounds
against six kinds of human cancer cell lines, including A549
(non-small-cell-lung cancer cell line), MDAMB-231 (human
breast cancer cell line), HepG2 (human liver carcinoma cell
line), HeLa (human epithelial cervical cancer cell line), HCT116
and RKO (human colon carcinoma cell line), were evaluated by
MTT assay, and aroylindoles (Fig. 1, compound B) was used as
the reference compounds. The results listed in Table 1 showed
that most of the synthesized compounds exhibited excellent
antiproliferative activity with IC50 values in the sub-micromol
level. Among the twelve chalcone derivatives, compound 12a
exhibited the highest activity with the IC50 values ranging from
23 to 79 nM, which implied that the chalcone derivative from
non-substituted indole-3-carbaldehyde is appropriate. Further
investigation of the structure-activity relationships showed
that the activity is related to the position of substituted groups
on the indole rings. Compound 12b exhibited slightly lower
activities than 12a indicating that N-methyl in the indole
moiety is unfavorable. This similar phenomenon also appeared
in other N-methyl and NH indole derivatives, for example, 12e
vs 12f and 12g vs 12h. However, 12c vs 12d exhibited the
Scheme 4. The synthesis of intermediate compound 21. Reagents and conditions: (i)
NBS, CH3CN, 2 h, 0°C, (a) HNO3, AcOH, 3 h, r.t. (b) Dimethylsulfate, K2CO3, acetone, 2d,
80°C. (c) Fe, AcOH, 8 h, r.t. (d) i: HCl, H2O, NaNO2, 30 min, -5°C, ii: NaOAc, 30 min, 0°C,
iii: KSeCN, 3 h, 0°C-r.t. (e) NaBH4, CH3I, EtOH, 2 min, r.t.
opposite trend. It is interesting that compound 25b
,
diarylketone derivatives derived from indole-4-carbaldehyde
and (5-bromo-2,3-dimethoxyphenyl)(methyl)selane, provided
the best results with IC50 values of 4 nM for A549 cell line, 6
nM for MM231, 13 nM for HEPG2, 22 nM for RKO, 21 nM for
HCT116, and 7 nM for HELA, respectively. The antiproliferative
activities of 25b are much more potent than reference
compound
A (IC50 values of 25 nM for A549 cell line, 33 nM for
MM231, 34 nM for HEPG2, 59 nM for RKO, 49 nM for HCT116,
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Table 2 Physical properties and prediction of absorption of 25b.
and 28 nM for HELA, respectively) , which suggested that the
introduction of selenium is beneficial to the antitumor activity.
The better activities than its N-CH3 analogue 27 also implied
that the non-substituted indole-4-carbaldehyde may be the
best choice for the preparation of this kind of tubulin
polymerization inhibitors. Furthermore, compounds 25a and
25c gave relatively lower activities than 25b, indicating that
the joint position of benzoyl on the indole ring is important.
Prediction of the absorption of compound 25b
Calculation a
MW
25b
374.3
3.87
Lipinski's rules
≤ 450
Clog P
≤ 5.0
HBA
HBD
3
1
≤ 10
≤ 5
PSA
47.56
0
≤ 90
Absorption levelb
In order to estimating the drug-like properties of selenium-
containing compound, we calculated Clog P, polar surface area
(PSA), hydrogen-bond acceptor atoms (HBA) and hydrogen-
bond donor atoms (HBD) of compound 25b. The absorption
level of 25b was also predicted with the ADMET program in
Discovery studio 2.1.21 The results showed in table 2 that
compound 25b meet the requirements of Lipinski's rules and
had a good absorption. The study of the PK profile in rats will
be performed in plan.
a
MW: molecular weight; clogP: calculated logarithm of the octanol-water partition
coefficient; HBA: hydrogen-bond acceptor atoms; HBD: hydrogen-bond donor atoms;
b
PSA: polar surface area; Data are predicted by discovery studio 2.1 soft (Accelrys). 0:
good absorption, 1: moderate absorption, 2 : low absorption, 3 : very low absorption.21
Effect of 25b on the inhibition of tubulin polymerization.
Numerous studies have demonstrated that the disruption of
microtubule dynamics induces the activation of mitochondrial
membrane potential complex and arrests the cell cycle in early
mitosis. To confirm the targeting of the tubulin-microtubule
system by the synthesized compounds, we chose 25b, which
exhibited the best antiproliferative activity in the MTT assay of
the six human cancer cell lines. As illustrated in Fig. 2, when
purified and unpolymerized tubulin was incubated at 37°C
(control sample), the increased fluorescence intensity with
time showed that tubulin polymerization had indeed occurred.
When tubulin was incubated with 25b at the specific
concentrations, the increased tendency of the fluorescence
intensity was obviously decreased with the increase of 25b
,
with an IC50 value of 2.1± 0.27 μM, which exhibited similar
activity with colchicine (IC50: 3.01 ± 0.12 μM). These data
indicated that 25b could effectively inhibit tubulin
polymerization and act as potential microtubule destabilizing
agents.
Fig. 2 In vitro tubulin polymerization assay. Tubulin in reaction buffer was incubated at
37°C in the absence (0.1% DMSO) or in the presence of 25b (10 μM, 5 μM, 2.5 μM, 1
μM, or 0.75 μM). The experiments were performed three times, and the results of
representative experiments are shown.
Effect of 25b on Cell Cycle Progression
It is known that most tubulin destabilizing agents exhibit the
ability to disrupt the regulated cell cycle distribution. As 25b
exhibited the most potent antiproliferative activities against
six kinds of human cancer cell lines, we then investigated the
effect of 25b on cell cycle distribution by flow cytometry
analysis. As shown in Fig. 3A, when HeLa cells were treated
with 1 to 10 nM 25b for 24 h, compared with the control
(DMSO), the accumulation of cells in the G2/M phase increased
sharply from 8.21% to 50.38%, with a concomitant decrease in
G1 and S phase cells. This trend was more obvious after the
incubation time was extended to 48 h (Fig. 3B). After the 48 h
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treatment, most of the cells were arrested at the G2/M phase late apoptosis cells were 5.89%, 13.47%, and 23.19%,
at relatively high concentrations (49.4% and 61.88% of G2/M respectively. The early and late apoptosis cells increased to
cell cycle arrest were found for 5 and 10 nM 25b treatments, 18.13%, 37.99%, and 71.33%, respectively, when the
Fig. 3B). Therefore, these results implied that 25b induced cell incubation time was extended to 72 h (Fig. 4B), which
cycle arrest at the G2/M phase.
indicated that 25b induced cell apoptosis in a concentration-
and time-dependent manner.
Mitochondrial dysfunction plays a vital role in the progression
of apoptosis. We therefore investigated the possible
involvement of mitochondrial dysfunction in 25b-induced
apoptosis of the cells by quantitative MMPs (mitochondrial
transmembrane potential) assay with JC-1 staining. As
expected (Fig. 4D), a rapid collapse of MMP was detected
when the HeLa cells were exposed to 5-10 nM of 25b for 48 h,
as a consequence of the opening of the permeability transition
pores that accumulate fluorescent dye from its red aggregated
to green-monomeric forms.
Effect of 25b on Cell Apoptosis Progression.
According to the cell cycle analysis results above, we
hypothesized that 25b treatment might induce the apoptosis
of HeLa cells. To evaluate this ability of compound 25b, flow
cytometry with propidium iodide (PI) and fluorescent
immunolabeling of the protein annexin-V (V-FITC) was
performed. After treating with various indicated
concentrations of 25b for 48 or 72 h, the HeLa cells were
harvested, stained with Annexin V-FITC and PI, and analyzed
by flow cytometry. It can be seen from Fig. 4A when the HeLa
cells were incubated with 25b at concentrations of 1, 5, and 10
nM or DMSO (0.01%) for 48 h, the percentages of early and
Table
1.
proliferative
activities of 12a
Anti-
–
12h, 12a, 12b, 13a,
13b, 14a, 14b, 25a
–
25c and 27
against human
cancer cell linesa.
Indole substitution
Entry
Comp.
12a
A549
MDAMB-231
HepG2
RKO
HCT116
HeLa
position
3
1
0.029 ± 0.002
0.041 ± 0.013
1.03 ± 0.034
0.344 ± 0.012
1.44 ± 0.096
1.26 ± 0.034
0.098 ± 0.013
0.113 ± 0.007
0.191 ± 0.013
0.051 ± 0.034
0.083 ± 0.056
0.079 ± 0.014
0.027 ± 0.007
0.004 ± 0.001
0.017 ± 0.007
0.063 ± 0.004
0.025 ± 0.003
0.038 ± 0.005
0.043 ± 0.002
1.41 ± 0.045
0.804 ± 0.023
0.445 ± 0.068
0.912 ± 0.066
0.068 ± 0.007
0.077 ± 0.005
0.218 ± 0.12
0.046 ± 0.002
0.055 ± 0.023
0.048 ± 0.017
0.038 ± 0.002
0.006 ± 0.0012
0.016 ± 0.007
0.087 ± 0.017
0.033 ± 0.011
0.023 ± 0.003
0.085 ± 0.012
4.14 ± 0.162
2.39 ± 0.134
0.314 ± 0.032
5.51 ± 0.177
0.115 ± 0.019
0.303 ± 0.015
1.14 ± 0.11
0.032 ± 0.001
0.054 ± 0.015
2.61 ± 0.098
1.14 ± 0.0019
1.03 ± 0.098
1.28 ± 0.098
0.095 ± 0.004
0.093 ± 0.014
0.223 ± 0.018
0.044 ± 0.001
0.079 ± 0.014
0.094 ± 0.004
0.074 ± 0.003
0.022 ± 0.007
0.052 ± 0.009
0.093 ± 0.089
0.059 ± 0.008
0.071 ± 0.002
0.087 ± 0.008
5.97 ± 0.035
3.56 ± 0.043
1.78 ± 0.025
6.78 ± 0.14
0.059 ± 0.002
0.065 ± 0.007
4.30 ± 0.056
1.86 ± 0.009
1.53 ± 0.078
3.49 ± 0.048
0.124 ± 0.005
0.288 ± 0.003
0.727 ± 0.07
0.037 ± 0.009
0.046 ± 0.002
0.086 ± 0.006
0.048 ± 0.009
0.007 ± 0.002
0.063 ± 0.007
0.050 ± 0.007
0.028 ± 0.002
2
12b
12c
3
3
3
3
3
3
3
4
4
5
5
3
4
5
4
4
3
4
12d
12e
12f
5
6
7
12g
12h
13a
13b
14a
14b
25a
25b
25c
0.095 ± 0.008
0.983 ± 0.006
1.85 ± 0.098
0.035 ± 0.019
0.054 ± 0.002
0.108 ± 0.017
0.089 ± 0.001
0.021 ± 0.003
0.076 ± 0.015
0.078 ± 0.004
0.049 ± 0.009
8
9
10
11
12
13
14
15
16
17
0.034 ± 0.014
0.044 ± 0.023
0.064 ± 0.018
0.056 ± 0.003
0.013 ± 0.005
0.046 ± 0.007
0.043 ± 0.006
0.034 ± 0.006
27
Comp. A
aCell lines were treated with compounds for 48 h. Cell viability was measured by MTT assay as described in the Experimental Section. bIC50 values are indicated as the
mean ± SD (standard error) of at least three independent experiments.
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Fig. 3 Effect of 25b on cell cycle progression. HeLa cells (3 × 105 cells/sample) were treated with specific concentrations of 25b for 24 h (A) or 48 h (B). Cells were harvested, fixed
with 70% ethanol, and stained with propidium iodide (PI). The cellular DNA content was then determined by flow cytometry.
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Fig. 4 Effect of 25b on cell apoptosis progression and decreased mitochondrial membrane potential of HeLa cells. Induction of apoptosis in the cultured HeLa cells upon treatment
with compound 25b. Dot plot representation of Annexin-V-FITC-fluorescence (x-axis) vs PI-fluorescence (y-axis) of the apoptotic HeLa (Annexin-V positive) cells, treated with
compound 25b (1, 5 and 10 nM) for 24 h (A) or 48 h (B). The HeLa cells were treated with 25b at different concentrations (1, 5, and 10 nM) or DMSO (0.01%) for 48 h, followed by
incubation with the fluorescence probe JC-1 for 30 min. Then, the cells were analyzed by fluorescence microscopy (D). The experiments were performed at least three times, and
the results of the representative experiments are shown
Effect of 25b on organization of cell microtubules
To study whether compound 25b could disrupt the
Conclusions
microtubule dynamics in living cells, we performed
immunofluorescence assay in HeLa cells as the tubulin-
microtubule system plays a vital role in the maintenance of cell
shape and basic cellular functions. As shown in Fig. 5, the
control group exhibited normal arrangement and organization
of the microtubule network in the HeLa cells, which were
characterized by regularly assembled, normal filiform
microtubules wrapped around the uncondensed cell nucleus.
In contrast, after exposure to 25b for 24 h, the spindle
formation demonstrated distinct abnormalities and was
heavily disrupted. Especially, when the concentration
increased to 10 nM, the microtubule spindles significantly
shrunk around the center of the cells, and a number of dotted
disorder formations were easily observed.
Considering that selenium is an essential trace mineral
nutrient with multiple roles in the growth and function of
living animal cells and it effectively inhibits tumorigenesis in
both animal models and epidemiological studies, in this paper,
we developed a series of selenium-containing chalcone and
diarylketone derivatives on the basis of previous studies. The
results of the antiproliferative activity assay in six human
cancer cell lines showed that most of these synthesized
compounds exhibited good to excellent activity. Among them,
the optimal compound 25b exhibited IC50 values of 0.004 to
0.022 μΜ for six cancer cell lines. Immunofluorescence assay
showed that 25b could effectively change the cell
microtubules organization in HeLa cells. The cell cycle
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progression in HeLa cells indicated that 25b induced G2/M
phase arrest, and cell apoptosis progression analysis showed
that this was associated with a collapse of mitochondrial
membrane potential (MMP). All of the above indicate that 25b
should be further studied for its drug abilities.
Notes and references
1
2
3
4
5
L. G. Wang, X. M. Liu, W. Kreis, D. R. Budman, Cancer
Chemoth. Pharm., 1999, 44, 355–36
S, Honore, E. Pasquier, D. Braguer, Cell. Mol. Life Sci., 2005,
62, 3039–3056.
T. Graening, H. G. Schmalz, Angew. Chem. Int. Ed., 2004, 43
3230−3256.
G. R. Pettit, S. B. Singh, E. Hamel, C. M. Lin, D. S. Alberts, D.
Garcia-Kendall, Experientia, 1989, 45, 209-211.
G. R. Pettit, S. B. Singh, M. R. Boyd, E. Hamel, R. K. Pettit, J.
M. Schmidt, F. Hogan, J. Med. Chem., 1995, 38, 1666-1672.
A. Brancale, R. Silvestri, Med. Res. Rev., 2007, 27, 209-238.
R. Kaur, G. Kaur, R. K. Gill, Eur. J. Med. Chem., 2014, 87, 89-
124.
.
Comparing to the reference compound A, a remarkable effect
of the oxygen by selenium substitution in 25b was exhibited.
Literatures have reported several mechanisms for the cancer
therapeutic effects of selenium derivatives including oxidative
stress, reduction of DNA damage, enhancement of the
immune response, inflammation, incorporation into
selenoproteins and so on. 25b showed a typical tubulin
polymerization inhibition responsible for the antiproliferative
activity and its other mechanisms of that are under further
study.
,
6
7
8
9
H. Chen, Y. Li, C. Sheng, J. Med. Chem., 2013, 56, 685-699.
B. L. Flynn, G. S. Gill, D. W. Grobelny, J. Med. Chem., 2011,
54, 6014-6027.
10 Y. S. Shan, J. Zhang, Z. Liu, Curr. Med. Chem., 2011, 18, 523-
538.
11 G. V. Kryukov, S. Castellano, S. V. Novoselov, A. V. Lobanov,
O. Zehtab, R. Guigó, V. N. Gladyshev, Science, 2003, 300
1439−1443.
12 C. Allmang, L. Wurth, Biochim. Biophys. Acta, 2009, 1790
1415−1423.
,
,
13 13. I. L. Clement, J. Donald Ganther, E. Howard, Anticancer
Res., 1998, 18, 4019-4025.
14 N. Nguyen, A. Sharma, N. Nguyen, A. K. Sharma, D. Desai, S.
J. Huh, S. Amin, C. Meyers, G. P. Robertson, Cancer Prev.
Res., 2011, 4, 248-58.
15 T. Lin, Z. Ding, N. Li, J. Xu, G. Luo, J. Liu, J. Shen, Eur. J. Cancer,
2011, 47, 1890-907.
16 C. Y. Chung, S. V. Madhunapantula, D. Desai, S. Amin, G. P.
Robertson, Cancer Prev. Res., 2011, 4, 935-48;
17 H. Zeng, W. H. Cheng, L. K. Johnson, J. Nutr. Biochem., 2013,
24, 776-80.
18 18. N. Chadha, O. Silakari, Eur. J. Med. Chem., 2017, 134
159-184.
,
19 J. P. Liou, C. Y. Wu, H. P. Hsieh, C. Y. Chang, C. M. Chen, C. C.
Kuo, J. Y. Chang, J. Med. Chem., 2007; 50, 4548-4552.
20 J. Yan, J. Chen, S. Zhang, J. Hu, L. Huang, X. Li, J. Med. Chem.,
2016, 59, 5264−5283.
21 42. W. J. Egan, K. M. Merz and J. J. Baldwin, J. Med. Chem.,
2000, 43, 3867-3877.
Fig. 5 Effects of 25b on spindle microtubules and chromosome organization of HeLa
cells. Cultured HeLa cells were incubated in the presence of varying concentrations of
25b (1, 5, 10 nM) for 12 h. The spindle microtubules were stained for
immunofluorescence using Alexa-Fluor 488 antibody (green) and corresponding
Hoechst 33342 conjugate (blue), and the chromosomes were stained with DAPI (blue).
Images were taken using an LSM 570 laser confocal microscope. The experiments were
performed three times, and the results of representative experiments are shown.
Acknowledgements
We thank the National Natural Science Foundation of China
(No. 21302235), Guangdong Natural Science Foundation
(2014A030313124), Guangdong High-level personnel of special
support program -Young top-notch talent project
(2015TQ01R244), Guangzhou Pearl River New Star Fund
Science And Technology Planning Project (201610010111) and
The Fundamental Research Funds for the Central Universities
(15ykpy04) for financial support of this study.
8 | J. Name., 2012, 00, 1-3
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