Design, synthesis and biological evaluation of benz-fused five-membered heterocyclic compounds as tubulin polymerization inhibitors with anticancer activities

Article abstract of DOI:10.1111/cbdd.13832

A series of benz-fused five-membered heterocyclic compounds were designed and synthesized as novel tubulin inhibitors targeting the colchicine binding site. Among them, compound 4d displayed the highest antiproliferative activity against four cancer cell lines with an IC₅₀ value of 4.9?μM in B16-F10 cells. Compound 4d effectively inhibited tubulin polymerization in vitro (IC₅₀ of 13.1?μM). Further, 4d induced cell cycle arrest in G2/M phase. Finally, 4d inhibited the migration of cancer cells in a dose-dependent manner. In summary, these results suggest that compound 4d represents a new class of tubulin inhibitors deserving further investigation.

Full text of DOI:10.1111/cbdd.13832

Received: 24 October 2020
DOI: 10.1111/cbdd.13832
Accepted: 31 January 2021
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R E S E A R C H A R T I C L E
Design, synthesis and biological evaluation of benz-fused five-
membered heterocyclic compounds as tubulin polymerization
inhibitors with anticancer activities
1
Yichang Ren1
Mingming Xue2
Binbin Cheng1
Jin Liu1
Buduma Komuraiah
|
|
|
|
|
Yao Liu3
Jianjun Chen1
|
1School of Pharmaceutical Sciences,
Guangdong Provincial Key Laboratory of
New Drug Screening, Southern Medical
University, Guangzhou, China
Abstract
A series of benz-fused five-membered heterocyclic compounds were designed and
synthesized as novel tubulin inhibitors targeting the colchicine binding site. Among
them, compound 4d displayed the highest antiproliferative activity against four
cancer cell lines with an IC50 value of 4.9 μM in B16-F10 cells. Compound 4d ef-
fectively inhibited tubulin polymerization in vitro (IC50 of 13.1 μM). Further, 4d
induced cell cycle arrest in G2/M phase. Finally, 4d inhibited the migration of cancer
cells in a dose-dependent manner. In summary, these results suggest that compound
4d represents a new class of tubulin inhibitors deserving further investigation.
2
Tianjin Tiancheng Chemical Co., Ltd,
Tianjin, China
3
Instrumental Analysis Center, Shanghai
Jiao Tong University, Shanghai, China
Correspondence
Yao Liu, Instrumental Analysis Center,
Shanghai Jiao Tong University, 800
Dongchuan Road, Shanghai 200240, China.
Email: yao_liu@sjtu.edu.cn
K E Y W O R D S
anticancer agents, CA-4 analogs, colchicine binding site, tubulin inhibitor
Jianjun Chen. School of Pharmaceutical
Sciences, Guangdong Provincial Key
Laboratory of New Drug Screening,
Southern Medical University, Guangzhou
5
10515, China.
Email: jchen21@smu.edu.cn
Funding information
Thousand Youth Talents Program (No.
C1080092); Scientific Research Project
of High Level Talents (No. G618319142)
in Southern Medical University of China;
International Science and Technology
Cooperation Projects of Guangdong
Province (No. G819310411)
1
|
INTRODUCTION
with high structural complexity which hampers further struc-
tural optimizations (Li et al., 2019; Lu et al., 2012; Vuuren
Out of the many clinically used antitumor drugs, microtubule
targeting agents (MTAs) (e.g., taxanes and vinca alkaloids)
have achieved considerable success in the treatment of var-
ious types of tumors such as melanoma and prostate cancer.
However, those tubulin-targeting agents are naturally derived
et al., 2015). Of the four types of tubulin inhibitors which
bind the taxane-, vinca alkaloid-, laulimalide- and colchicine-
site, colchicine binding site inhibitors (CBSI) are generally
more amenable to modifications due to the relative structural
simplicity as compared to the other three types of MTAs
Buduma Komuraiah and Yichang Ren contributed equally.
Chem Biol Drug Des. 2021;97:1109–1116.
wileyonlinelibrary.com/journal/cbdd
© 2021 John Wiley & Sons A/S.
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1109

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KOMURAIAH et Al.
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with a natural origin (Bai et al., 1990; Bennett et al., 2012;
Bollag et al., 1995; Kingston, 2009; Risinger et al., 2009;
Wei et al., 2004). Therefore, the development of CBSIs has
been intensified in the past two decades with numerous novel
and potent CBSIs being discovered (Figure 1), for example,
ATCAA (Lu et al., 2009), AICA (Lu et al., 2010), SMART (Lu
et al., 2011), PAT (Lu et al., 2014), ABI-I (Chen et al., 2010),
ABI-II (Chen et al., 2011), ABI-III (Chen et al., 2012), and
RABI (Xiao et al., 2013).
We previously discovered a series of 2-aryl-4-benzoyl-
imidazole (ABI-II) analogs as antiproliferative agents targeting
the colchicine binding site in tubulin for treating melanoma and
prostate cancer. The ABI-II analogs were designed based on the
first generation ABI-I analogs utilizing a ring-fusion strategy,
namely, fusing the A- and B-ring of ABI-I (Figure 1). However,
we only synthesized a very limited number of compounds (<5)
that showed moderate antiproliferative activities with IC in
phenol (1, X = O). Initially, the nitro group was reduced by
H in the presence of Pd/C to obtain the amine derivatives (2,
2
X = O) which were converted to benzoxazoles (3, X = O)
o
using triethyl orthoformate in the presence of 3A molecular
sieves (MS) at 130°C (Scheme 1, step b). The same synthetic
strategy was applied to the synthesis of substituted thiophenol
derivatives (Scheme 1, step b). The synthesis of compound 4
(a-j) was achieved by oxidative C-H activation at the C-2 po-
sition of benzoxazole or benzothiazole with 3,4.5-trimethoxy
benzaldehyde using ammonium persulphate and tetrabutyl
ammonium bromide (Siddaraju et al., 2014) as a phase trans-
fer catalyst (Scheme 1, step c). Compound 5f was prepared
through Suzuki coupling by reacting 4f with 3-fluoro-4-meth
oxy-phenylboronic acid (Scheme 1, step d).
The synthesis of series 2 compounds [1-(3,4,5-trimethox
ybenzoyl)-benzimidazoles, 10a-10f)] is shown in Scheme 2.
Briefly, the substituted 1,2-diaminobenzene 8 was re-
acted with ethyl formate to generate the benzimidazole 9.
Compound 9 was acylated by 3,4.5-trimethoxy benzaldehyde
at the N-1 position to give the desired 1-(3,4,5-trimethoxybe
nzoyl)-benzimidazoles 10a-10f.
In addition to the 1-(3,4,5-trimethoxybenzoyl)-benzim
idazole compounds 10a-10f, we also synthesized series 2
compounds with the 3,4,5-trimethoxybenzoyl moiety at the
1-position of the benz-, pyridine or pyrimidine-fused five-
membered heterocycles, as shown in Scheme 3.
50
the micromolar range (e.g., ~10 μM). To extend the structure-
activity relationship study of this scaffold, we designed two
series of target compounds (Benz-fused five-membered hetero-
cyclic compounds, Figure 2) based on the following rationales:
1
) incorporating different five-membered heterocyclic B-ring
(
e.g., thiazole, oxazole, pyrrole, triazole) to understand the toler-
ability of the B-ring modifications (Series 1), as benzimidazole
and benzothiazole moieties are found in many existing tubulin
inhibitors (Ashraf et al., 2016; Fu et al., 2020). 2) shifting the
position of the 3,4,5-trimethoxyphenyl (TMP) group from 2 to
1
to explore the effects of different geometric configurations
(
Series 2); With the TMP moiety at position-1, series 2 is es-
2.2
|
Biological evaluation
sentially a structure mimic of CA-4, which is a highly potent
CBSI in clinical trials. Herein, we describe the synthesis and
biological evaluation of these compounds as detailed in the fol-
lowing section.
2.2.1
|
In vitro antiproliferative activity and
structure-activity relationship
The antiproliferative activities of the newly synthesized
compounds were evaluated against four cancer cell lines
(MCF-7, H1299, HeLa and B16-F10) by an MTT assay
with CA-4 and colchicine as the positive controls. As pre-
sented in Table 1, most of the compounds are not very ac-
tive with IC > 10 μM. Compounds 4d, 4f, 4g exhibited
2
2
|
RESULTS AND DISCUSSION
Chemistry
.1
|
5
0
As outlined in Scheme 1, the synthesis of series 1 compounds
started with commercially available 4- or 5-substituted 2-nitro
moderate potency with IC ranging from 4.9 to 10 μM.
50
In general, the benzothiazole analogs (e.g., 4d-4g, IC of
5
0
FIGURE 1 Structures of known
CBSIs [Colour figure can be viewed at
wileyonlinelibrary.com]

KOMURAIAH et Al.
1111
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4
.9~18.6 μM) exhibited slightly higher potency than that
with R as a hydrogen (4b vs. 4a; 4e vs. 4d; 4i vs. 4h, 10b
2
of the benzoxazole (e.g., 4a-c, IC of 16.5 ~ 46.2 μM) and
vs. 10a; 10d vs. 10c). Also, the geometrically “extended”
Series 1 compounds are slightly better than their geometri-
cally “bent” counterparts Series 2 compounds (4b vs. 10f;
4f vs. 10a; 4d vs. 10c; 4e vs. 10d). Specifically, for se-
ries 1 compounds, converting the oxazole (4a) to thiazole
5
0
benzimidazole (e.g., 10a-f, IC of 12.58~46.8 μM) com-
5
0
pounds. For the R and R substituents on the benz-fused
1
2
ring systems, when R is a hydrogen, the compounds are
1
generally less potent than that of the corresponding analogs
(
4g) led to improved potency. Compounds with electron-
withdrawing groups on the benzothiazole ring displayed
higher potency than the ones with electron-donating groups
(
e.g., 4d, 4f, 4g and 4h; -Br>-Cl>-OCH >-NH ). Among
3 2
them, compound 4d (6-Br benzothiazole) exhibited the
highest activity with an average IC value of 7.219 μM.
5
0
The 6-substituted benzothiazole derivatives are less potent
than that of 5-substituted benzothiazole derivatives [4d
(
IC = 7.219 μM) vs. 4e (IC > 10 μM)]. These results
50 50
indicated that the 6-Br benzothiazole moiety is optimal for
activity. Series 2 compounds (10a-f, 11–15) showed low ac-
tivities (IC > 10 μM), which might be due to the shift of the
5
0
position of C-Ring (3,4,5-trimethoxy benzoyl group) from
2
4
-position to 1-position of the imidazole ring. Compound
j with a N-methyl indolyl group as C-ring showed essen-
tially a total loss of activity (IC > 10 μM), comparing to
FIGURE 2 Rationale for the design of target compounds [Colour
figure can be viewed at wileyonlinelibrary.com]
50
compound 4f, suggesting that the 3,4,5-trimethoxypheny
SCHEME 1 Synthesis of series 1 compounds
SCHEME 2 Synthesis of series 2 compounds (10a-10f)

1
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KOMURAIAH et Al.
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SCHEME 3 Synthesis of series 2 compounds (11–15) [Colour figure can be viewed at wileyonlinelibrary.com]
play an important role in maintaining the cytotoxic activity.
The positions of substituents on the benz-fused rings (10a,
indicated that compound 4d inhibited tubulin polymeri-
zation with an IC of 13.1 μM (Figure 3a), weaker than
5
0
1
0c vs. 10b, 10d) do not affect the potencies, but electron-
that of CA-4 (IC = 1.84 μM, Figure 3b) and colchicine
50
withdrawing groups (10a-d, 11, 12) seem to be better than
that of electron-donating substituents (10e-f, 13, 14). In ad-
dition, the cytotoxicity of these compounds was examined
against a normal cell line HUVEC (human umbilical vein
endothelial cell). As shown in Table 1, all the compounds
demonstrated little toxicity to HUVEC with IC > 50 μM,
(IC = 7.15 μM, Figure 3c). Furthermore, compound 4d
50
inhibited tubulin polymerization in a dose-dependent man-
ner (Figure 3d).
2.2.3
|
Immunofluorescence studies
5
0
suggesting that these compounds cause selective toxicity
to cancer cells over normal cells. Interestingly, similar
benzimidazole and benzothiazole analogs have been re-
ported (Ashraf et al., 2016; Fu et al., 2020), but our com-
pounds showed slightly lower potencies probably due to
a higher fraction of drugs being transported out of cells
by efflux pumps (Figure S55). Also the newly synthesized
compounds are generally less potent than that of CA-4
and of the natural ligand colchicine, partially because of
the lower binding affinities in the colchicine binding site
of tubulin, as seen from the molecular docking studies of
colchicine and compound 4d (Figure S54). However, the
advantages of these compounds may be the higher stability
It is known that microtubule dynamics play an important
role in cancer cell growth. To validate whether compound
4d might influence microtubule dynamics, an immunofluo-
rescent assay in B16-F10 cells was performed. As showed
in Figure 4, in the control group, the microtubule networks
exhibited a normal arrangement with slim and fibrous mi-
crotubules (green) wrapped around the cell nucleus (blue).
However, when cells were exposed to compound 4d (1, 5
and 10 μM) or CA-4 (10 nM) for 6 hr, the microtubule
networks were disrupted in comparison with the control.
These results suggest that compound 4d might induce the
collapse of the microtubule networks in a dose-dependent
manner.
(
Figure S57) over CA-4 because there will be no cis-trans
conversion due to the presence of a double bond as in the
case of CA-4.
Cells were exposed to different concentrations of com-
pounds for 48 hr to determine the cell viability through MTT
assay. IC values are presented as the mean ± SD of at least
2.2.4
|
Cell cycle study by flow cytometry
Tubulin polymerization inhibitors have been previously
known to impact cell division. Thus, we evaluated the effects
of compound 4d on the cell cycle of B16-F10 cells with flow
cytometry. As shown in Figure 5, compound 4d arrested cell
cycle at G2/M phase (Figure 5a) in a dose-dependent man-
ner. When B16-F10 cells were exposed to increasing con-
centrations of compound 4d (1, 5 and 10 μM), the percentage
of cells in the G2/M phase was significantly increased from
8.27% to 82.90%.
5
0
three independent experiments.
2
.2.2
|
In vitro tubulin polymerization assay
To explore the mechanism of action, compound 4d was
chosen to evaluate the effects on tubulin polymerization
with CA-4 and colchicine as positive controls. The results

KOMURAIAH et Al.
1113
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TABLE 1 In vitro antiproliferative activity of series 1 and 2 compounds
IC50 (μM)
Structure
Compound
X
R1
R2
MCF−7
H1299
HeLa
B16-F10
HUVEC
4
4
4
4
4
4
4
4
4
4
a
b
c
d
e
f
g
h
i
O
O
O
S
S
S
S
S
S
OCH3
H
CH3
Br
H
Cl
OCH3
NH2
H
H
OCH3
H
H
Br
H
H
H
NH2
21.58 ± 1.23
32.85 ± 2.51
46.18 ± 1.96
9.450 ± 0.292
16.79 ± 0.35
10.00 ± 0.14
9.253 ± 0.065
25.47 ± 1.36
50.25 ± 3.87
16.20 ± 0.68
24.60 ± 0.98
44.12 ± 3.41
38.66 ± 0.82
7.652 ± 0.215
18.62 ± 0.54
6.761 ± 0.171
6.538 ± 0.152
32.18 ± 2.17
61.24 ± 5.66
13.88 ± 0.60
19.58 ± 0.47
27.65 ± 1.87
29.78 ± 2.54
6.862 ± 0.144
12.85 ± 0.40
7.605 ± 0.238
15.72 ± 0.28
22.07 ± 0.66
39.87 ± 3.25
19.67 ± 1.02
16.49 ± 1.05
30.17 ± 0.69
30.06 ± 3.81
4.912 ± 0.088
21.39 ± 0.63
7.273 ± 0.411
13.14 ± 0.32
19.03 ± 0.25
41.08 ± 2.96
13.56 ± 0.24
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
j
5
f
32.90 ± 1.94
38.62 ± 1.03
41.37 ± 1.26
22.53 ± 1.97
>50
10a
10b
10c
10d
10e
10f
11
Cl
H
Br
H
H
H
H
Cl
H
Br
CH3
OCH3
13.77 ± 0.51
17.22 ± 0.62
12.58 ± 1.21
15.29 ± 0.68
35.68 ± 3.76
29.60 ± 2.33
18.30 ± 4.51
24.18 ± 0.85
27.17 ± 3.15
20.73 ± 2.30
32.08 ± 1.59
52.12 ± 4.63
46.97 ± 1.26
27.24 ± 0.81
31.21 ± 2.09
18.96 ± 0.67
25.36 ± 1.71
22.14 ± 0.96
45.39 ± 2.64
39.85 ± 1.80
34.12 ± 2.07
19.84 ± 2.45
22.68 ± 0.55
17.52 ± 1.37
18.81 ± 1.09
38.47 ± 2.35
33.46 ± 2.11
23.59 ± 0.58
>50
>50
>50
>50
>50
>50
>50
1
2
20.88 ± 2.01
19.05 ± 1.70
24.86 ± 2.31
30.17 ± 1.06
>50
1
1
1
3
4
5
32.90 ± 3.14
69.17 ± 3.61
55.28 ± 7.23
25.83 ± 0.77
72.64 ± 2.08
69.40 ± 4.28
36.10 ± 1.27
48.19 ± 2.15
74.32 ± 3.49
45.92 ± 3.32
67.83 ± 6.18
60.12 ± 2.37
>50
>50
>50
CA-4
Colchicine
0.013 ± 0.002
0.024 ± 0.006
0.014 ± 0.002
0.120 ± 0.016
0.009 ± 0.005
0.045 ± 0.004
0.007 ± 0.001
0.068 ± 0.012
>50
>50
2
.2.5
|
Inhibition of cancer cell migration
organs and microtubule play an important role in cell mi-
gration. A wound healing assay was performed in order to
confirm the effects of compound 4d in cell migration. When
Knowing that cancer cells are able to migrate to distant

1
114
KOMURAIAH et Al.
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FIGURE 3 Inhibition of tubulin polymerization in vitro. (a) The inhibition of tubulin polymerization by compound 4d. (b) The inhibition
of tubulin polymerization by CA-4. (c) The inhibition of tubulin polymerization by colchicine. (d) Compound 4d exhibited a dose-dependent
inhibition of tubulin polymerization [Colour figure can be viewed at wileyonlinelibrary.com]
FIGURE 4 Effects of compound 4d on microtubules. B16-F10 cells were treated with vehicle control 0.1% DMSO (a), compound 4d (1 μM)
(
(
b), compound 4d (5 μM) (c), compound 4d (10 μM) (d), CA-4 (10 nM) (e) for 6 hr. Microtubules were visualized with an anti-β-tubulin antibody
green), and the cell nucleus was visualized with DAPI (blue). Fluorescence images were collected by LSM 880 laser confocal microscope (Carl
Zeiss, Germany) [Colour figure can be viewed at wileyonlinelibrary.com]
treating the B16-F10 cells with 1, 5 and 10 μM of compound
antiproliferative potency with IC values larger than
50
4
d, the wound closure was potently suppressed (Figure 6),
10 μM. Several of them (4d, 4f, 4g) demonstrated moderate
potency against a panel of cancer cell lines with IC val-
showing that the migration of cancer cells was inhibited by
compound 4d in a dose-dependent manner.
5
0
ues in the micromolar range (e.g., 4.7–10 μM). Structure-
activity relationships revealed that the relative position of
the 3,4,5-trimethoxybenzoyl group on the five-membered
heterocyclic ring has significant influence on the biological
activities of the compounds, with 2-position being optimal.
Among the newly synthesized compounds, 4d displayed
the highest antiproliferative activity against four cancer cell
lines in vitro. In addition, compound 4d was able to inhibit
tubulin polymerization in vitro in a dose-dependent manner.
3
|
CONCLUSION
In summary, a series of benz-fused five-membered hetero-
cyclic compounds were designed and synthesized as tubu-
lin polymerization inhibitors and a focused SAR study
was performed. Most of these compounds exhibited low

KOMURAIAH et Al.
1115
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FIGURE 5 Cell cycle arrested by compound 4d. (a) Compound 4d induced G2/M arrest in B16-F10 cells. B16-F10 cells were incubated with
varying concentrations of compound 4d (1, 5 and 10 μM) and CA-4 (10 nM) for 48 hr. The percentages of cells in different phases of the cell cycle
were analyzed by FlowJo 7.6.1. (b) Histograms displayed the percentage of cell cycle distribution after treatment with compound 4d and CA-4
[
Colour figure can be viewed at wileyonlinelibrary.com]
FIGURE 6 Effects on the B16-F10 cells migration. (a) Representative images of wound area in a wound healing assay. Images were obtained
at 0 hr and 24 hr after treatment with 0, 1, 5 and 10 μM compound 4d and 10 nM CA-4. (b) Histograms displayed the length of the scratches at 0 hr
and 24 hr. (n = 3, **p <.05 vs. DMSO control) [Colour figure can be viewed at wileyonlinelibrary.com]
Further mechanism studies indicated that compound 4d in-
duced cell cycle arrest in G2/M phase. Finally, compound
compounds 10c and 11 at various time points in cell media (with-
out cells) were determined by HPLC (Figure S57, Figure S58).
4
d inhibited the migration of cancer cell in a dose-dependent
manner. Taken together, these results suggest that compound
d represents a promising tubulin inhibitor deserving further
investigation.
ACKNOWLEDGEMENT
4
This work was supported by scientific research project of
high level talents (No. G618319142) in Southern Medical
University of China; Thousand Youth Talents Program
(
No. C1080092) from the Organization Department of the
3
.1
|
Supporting information availability
CPC Central Committee, China; International Science and
Technology Cooperation Projects of Guangdong Province
(No. G819310411).
1
13
The supporting information contains H NMR, C NMR and
mass spectra (Figure S1 to Figure S50). The purity of com-
pounds 4d, 4f and 4g was determined by HPLC (Figure S51 to
Figure S53). Molecular modeling of colchicine and compound
ORCID
Yao Liu
https://orcid.org/0000-0002-9922-7509
4
d at colchicine binding site of tubulin was shown in Figure S54.
The concentrations of compounds 10c and 11 at various time
points in cell media (with cells co-cultured) were determined
by HPLC (Figure S55, Figure S56). The concentrations of
REFERENCES
Ashraf, M., Shaik, T. B., Malik, M. S., Syed, R., Mallipeddi, P. L.,
Vardhan, M., & Kamal, A. (2016). Design and synthesis of

1
116
KOMURAIAH et Al.
|
cis-restricted benzimidazole and benzothiazole mimics of combret-
astatin A-4 as antimitotic agents with apoptosis inducing ability.
Bioorganic and Medicinal Chemistry Letters, 26, 4527–4535.
Lu, Y., Li, C. M., Wang, Z., Chen, J., Mohler, M. L., Li, W., Dalton,
J. T., & Miller, D. D. (2011). Design, synthesis, and SAR studies
of 4-substituted methoxylbenzoyl-aryl-thiazoles analogues as po-
tent and orally bioavailable anticancer agents. Journal of Medicinal
Chemistry, 54, 4678–4693.
Lu, Y., Li, C. M., Wang, Z., Ross, C. R. 2nd, Chen, J., Dalton, J.
T., Li, W., & Miller, D. D. (2009). Discovery of 4-substituted
methoxybenzoyl-aryl-thiazole as novel anticancer agents: Synthesis,
biological evaluation, and structure-activity relationships. Journal
of Medicinal Chemistry, 52, 1701–1711.
Lu, Y., Wang, Z., Li, C. M., Chen, J., Dalton, J. T., Li, W., & Miller, D.
D. (2010). Synthesis, in vitro structure-activity relationship, and in
vivo studies of 2-arylthiazolidine-4-carboxylic acid amides as anti-
cancer agents. Bioorganic and Medicinal Chemistry, 18, 477–495.
Risinger, A. L., Giles, F. J., & Mooberry, S. L. (2009). Microtubule
dynamics as a target in oncology. Cancer Treatment Reviews, 35,
255–261.
Siddaraju, Y., Lamani, M., & Prabhu, K. R. (2014). A transition metal-
free Minisci reaction: Acylation of isoquinolines, quinolines, and
quinoxaline. Journal of Organic Chemistry, 79, 3856–3865.
van Vuuren, R. J., Visagie, M. H., Theron, A. E., & Joubert, A. M.
(2015). Antimitotic drugs in the treatment of cancer. Cancer
Chemotherapy and Pharmacology, 76, 1101–1112.
Bai, R. L., Pettit, G. R., & Hamel, E. (1990). Binding of dolastatin
1
0 to tubulin at a distinct site for peptide antimitotic agents near
the exchangeable nucleotide and vinca alkaloid sites. Journal of
Biological Chemistry, 265, 17141–17149.
Bennett, M. J., Chan, G. K., Rattner, J. B., & Schriemer, D. C.
(
2012). Low-dose laulimalide represents a novel molecular
probe for investigating microtubule organization. Cell Cycle, 11,
045–3054.
3
Bollag, D. M., McQueney, P. A., Zhu, J., Hensens, O., Koupal, L.,
Liesch, J., Goetz, M., Lazarides, E., & Woods, C. M. (1995).
Epothilones, a new class of microtubule-stabilizing agents with a
taxol-like mechanism of action. Cancer Research, 55, 2325–2333.
Chen, J., Ahn, S., Wang, J., Lu, Y., Dalton, J. T., Miller, D. D., & Li,
W. (2012). Discovery of novel 2-aryl-4-benzoyl-imidazole (ABI-
III) analogues targeting tubulin polymerization as antiproliferative
agents. Journal of Medicinal Chemistry, 55, 7285–7289.
Chen, J., Li, C. M., Wang, J., Ahn, S., Wang, Z., Lu, Y., Dalton, J. T.,
Miller, D. D., & Li, W. (2011). Synthesis and antiproliferative activity
of novel 2-aryl-4-benzoyl-imidazole derivatives targeting tubulin po-
lymerization. Bioorganic and Medicinal Chemistry, 19, 4782–4795.
Chen, J., Wang, Z., Li, C. M., Lu, Y., Vaddady, P. K., Meibohm, B.,
Dalton, J. T., Miller, D. D., & Li, W. (2010). Discovery of novel
Wei, W., Ayad, N. G., Wan, Y., Zhang, G. J., Kirschner, M. W., &
Kaelin, W. G. Jr (2004). Degradation of the SCF component Skp2
in cell-cycle phase G1 by the anaphase-promoting complex. Nature,
428, 194–198.
Xiao, M., Ahn, S., Wang, J., Chen, J., Miller, D. D., Dalton, J. T., &
Li, W. (2013). Discovery of 4-Aryl-2-benzoyl-imidazoles as tubu-
lin polymerization inhibitor with potent antiproliferative properties.
Journal of Medicinal Chemistry, 56, 3318–3329.
2
-aryl-4-benzoyl-imidazoles targeting the colchicines binding site
in tubulin as potential anticancer agents. Journal of Medicinal
Chemistry, 53, 7414–7427.
Fu, D. J., Liu, S. M., Li, F. H., Yang, J. J., & Li, J. (2020). Antiproliferative
benzothiazoles incorporating a trimethoxyphenyl scaffold as novel
colchicine site tubulin polymerisation inhibitors. Journal of Enzyme
Inhibition and Medicinal Chemistry, 35, 1050–1059.
Kingston, D. G. (2009). Tubulin-interactive natural products as antican-
cer agents. Journal of Natural Products, 72, 507–515.
Li, L., Quan, D., Chen, J., Ding, J., Zhao, J., Lv, L., & Chen, J. (2019).
Design, synthesis, and biological evaluation of 1-substituted -2-aryl
imidazoles targeting tubulin polymerization as potential anticancer
agents. European Journal of Medicinal Chemistry, 184, 111732.
Lu, Y., Chen, J., Wang, J., Li, C. M., Ahn, S., Barrett, C. M., Dalton,
J. T., Li, W., & Miller, D. D. (2014). Design, synthesis, and biolog-
ical evaluation of stable colchicine binding site tubulin inhibitors
as potential anticancer agents. Journal of Medicinal Chemistry, 57,
SUPPORTING INFORMATION
Additional supporting information may be found online in
the Supporting Information section.
How to cite this article: Komuraiah B, Ren Y, Xue M,
et al. Design, synthesis and biological evaluation of
benz-fused five-membered heterocyclic compounds as
tubulin polymerization inhibitors with anticancer
activities. Chem Biol Drug Des. 2021;97:1109–1116.
https://doi.org/10.1111/cbdd.13832
7
355–7366.
Lu, Y., Chen, J., Xiao, M., Li, W., & Miller, D. D. (2012). An overview
of tubulin inhibitors that interact with the colchicine binding site.
Pharmaceutical Research, 29, 2943–2971.