Synthesis and evaluation of novel α-substituted chalcones with potent anti-cancer activities and ability to overcome multidrug resistance

Article abstract of DOI:10.1016/j.bioorg.2019.03.014

A series of forty α-substituted chalcones were synthesized and screened for their antiproliferative activities against HCT116 (colorectal) and HCC1954 (breast) cancer cell lines. Compounds 5a and 5e were found to be the most potent compounds with GI₅

Full text of DOI:10.1016/j.bioorg.2019.03.014

Accepted Manuscript
Synthesis and evaluation of novel α-substituted chalcones with potent anti-can-
cer activities and ability to overcome multidrug resistance
Sharon Riaz, Maheen Iqbal, Rahim Ullah, Rida Zahra, Ghayoor Abbas Chotana,
Amir Faisal, Rahman Shah Zaib Saleem
PII:
S0045-2068(19)30026-4
DOI:
https://doi.org/10.1016/j.bioorg.2019.03.014
YBIOO 2840
Reference:
Bioorganic Chemistry
To appear in:
Received Date:
Revised Date:
Accepted Date:
7 January 2019
24 February 2019
6 March 2019
Please cite this article as: S. Riaz, M. Iqbal, R. Ullah, R. Zahra, G. Abbas Chotana, A. Faisal, R. Shah Zaib Saleem,
Synthesis and evaluation of novel α-substituted chalcones with potent anti-cancer activities and ability to overcome
multidrug resistance, Bioorganic Chemistry (2019), doi: https://doi.org/10.1016/j.bioorg.2019.03.014
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Synthesis and evaluation of novel α-substituted chalcones with potent anti-
cancer activities and ability to overcome multidrug resistance.
Sharon Riaz,a Maheen Iqbal,b Rahim Ullah,b Rida Zahra,b Ghayoor Abbas Chotana,a Amir
Faisal*,b, Rahman Shah Zaib Saleem*,a
a Department of Chemistry and Chemical Engineering, Syed Babar Ali School of Science and
Engineering, Lahore University of Management Sciences, Lahore, 54792, Pakistan.
b Department of Biology, Syed Babar Ali School of Science and Engineering, Lahore
University of Management Sciences, Lahore, 54792, Pakistan.
* Corresponding authors: Rahman Shah Zaib Saleem (rahman.saleem@lums.edu.pk) and
Amir Faisal (amir.faisal@lums.edu.pk)
Abstract
A series of forty α-substituted chalcones were synthesized and screened for their
antiproliferative activities against HCT116 (colorectal) and HCC1954 (breast) cancer cell
lines. Compounds 5a and 5e were found to be the most potent compounds with GI50 values of
0.63 µM and 0.725 µM in HCC1954 cell line and 0.69 µM and 1.59 µM in HCT116 cell line,
respectively. Both compounds induced a G2/M cell cycle arrest and caused apoptotic cell
death in HCT116 cells as shown by the induction of PARP cleavage. The compounds also
stabilized p53 in a dose-dependent manner in HCT116 cells following 24-hour treatment.
Furthermore, both 5a and 5e were able to overcome multidrug resistance in two MDR-1
overexpressing multidrug resistant cell lines.
Keywords: α-substituted chalcones; antiproliferative activity; apoptotic activity; cell cycle
arrest; multidrug resistance;
1. Introduction
Cancer is a large group of heterogeneous diseases characterized by abnormal division and
spread of cells.1 Chemotherapy, surgery, radiotherapy or their combinations are the current
modes of cancer treatment.2 Among these, systemic chemotherapy is one of the most
common and effective treatments for many types of cancer.3 However, the development of
multidrug resistance (MDR) is a major impediment to the success of chemotherapy.4 It is,
therefore, important to design and discover new drugs with anticancer potential and ability to
overcome MDR.
Chalcones (α, β-unsaturated aromatic ketones) are medicinally important compounds.5
Various natural and synthetic chalcones have been shown to display biological properties to
act as potential hits for inflammation,6,7 diabetics,8 Alzheimer’s disease,9 viral diseases10 and
anti-cancer11-15 activities. Naturally occurring chalcones such as Isoliquiritigenin and
Licochalcone A arrest cells in the S and G2/M phase of the cell cycle.16 Synthetic chalcones
such as compounds (i–v) have been shown to overcome Pgp-mediated MDR.17 Chalcones
have gained attention in the recent literature for pharmaceutical5 and photochemical
applications.18 Recently, a number of chalcone derivatives have been shown to have
anticancer potential. FC77 has been described as a potential agent against MDR
1

malignancies.19 Compound vi, a thienopyrimidine−chalcone hybrid, showed anticancer
activity against a breast cancer cell line MCF-7, with GI50 value of 6.52 ± 0.82 μM.20
Compound vii inhibited cell proliferation by arresting the cells at the G2/M phase in
HCCT116 cell line.21 Similarly benzo[d]imidazo[2,1-b]thiazole-chalcone conjugates (e.g.,
compound (viii)) caused a G2/M cell cycle arrest in a human breast cancer cell line (MDA
MB-231).22 Compound (ix) displayed a GI50 value of 2.3 ± 0.3 μM in NCI-H460 cell line and
acted through ROS modulation that induced caspase-3 mediated apoptosis.23 Novel
Indolizine-chalcone hybrids (x) showed caspase-dependent apoptosis of human Lymphoma
U937 cells with GI50 of 1.12 μM.24 Carbazole derivatives containing chalcone analogues
(CDCAs), e.g., compound (xi), were found to be apoptotic inducers (GI50 = 0.22 μM in
human acute leukemia cell line HL-60).25 DK-139 was found to induce caspase-mediated
apoptosis through unfolded protein response in lung cancer-A549 cell line.26 Chalcones such
as T37 were reported to activate p53.27 Similarly, compounds D14 and D15 suppress
migration and invasion of osteosarcoma cells through p53 regulation.28
Figure 1 Natural and synthetic chalcones and examples from recent literature (2018)
Despite abundant literature on the chalcones, lesser is known about the activities of the alpha-
substituted chalcones. The literature cites only a handful of such compounds, for example, α-
Alkyl, α-COOEt, α-COOH, α-CN, α-triazole, α-halo substitutions and their biological
profiling have been reported by Duchi,29 Lawrence,30 Smith31 and Yin32. We have been
interested in secondary metabolites33-37 as well as in elucidating the bioactivities of small
organic molecules.38-44 Recently, we have reported two chalcone derivatives, SSE14105 and
2

SSE14106 that can stabilize p53-tumor suppressor protein in HCT116 cells and inhibit their
proliferation.38 We have also identified the compound SSE15206, a pyrazolinethioamide
derivative of SSE14108, to be a microtubule depolymerizing agent that could overcome
MDR.39
Figure 2 Anti-tumor chalcones from our previous work.
In continuation of our interest in probing the anticancer activities of chalcones, we herein
report synthesis and evaluation of anticancer activity of 40 α-substituted chalcones (including
37 novel compounds) that contain α-aryloxy, α-COOEt, α-COOH and α-amide groups. We
have determined their antiproliferative activity against HCT116 (colorectal) and HCC1954
(breast) cancer cell lines. Moreover, we have evaluated the most active compounds (5a and
5e) for their ability to induce cell cycle arrest, p53 stabilization, apoptosis and to overcome
multidrug resistance. To our knowledge, this is the first report of such molecules overcoming
multidrug resistance.
2. Results and Discussion
2.1 Chemistry
2.1.1 Synthesis of α-aryloxy chalcones
We synthesized series of α-2-naphthalen-2-yloxy, α-3,5-dimethyl phenoxy and α-1,3-
benzodioxol-5-yloxy substituted chalcones as shown in Scheme 1:
3

Scheme 1. Synthesis of α-Aryloxy chalcones. Reagents and conditions: (i) K2CO3, ACN,
r.t., 48 hr.; (ii) ArCHO, KOH, EtOH, r.t., 24 hr.
2.1.1.1
Synthesis of α-2-(naphthalen-2-yloxy)-chalcone (2a – 2s)
In order to prepare α-2-(naphthalen-2-yloxy)-chalcone, 2-naphthol was reacted with 2ˊ-
bromoacetophenone using Williamson-ether synthesis.45 The resulting 2-(naphthalen-2-
yloxy)-1-phenylethanone (2) was then condensed with different aldehydes through Claisen-
Schmidt condensation leading to the formation of α-2-(naphthalen-2-yloxy)-chalcone
derivatives (2a - 2s).
Table 1a:Structure, yields and inhibition of cell proliferation in HCC1954 and HCT116
cell lines.
Ar1
R1
R2
R3
R4
Yield
(%)
HCC195
HCT116
GI50 (µM) ± SD
GI50 (µM) ± SD
-
H
H
H
Cl
H
H
H
H
H
H
NO2
H
H
H
H
H
H
H
H
H
H
F
85
47
40
55
74
47
34
18
13
23
41
78
69
73
11
>100 (N = 2)
>100 (N = 2)
>100 (N = 3)
>100 (N = 3)
2a
2b
2c
2d
2e
2f
-
OMe
-
Cl
23.54 ± 0.18 (N = 2) 19.35 ± 2.56 (N = 3)
64.26 ± 2.89 (N = 2) 43.98 ± 2.71 (N = 3)
-
H
-
Me2CH
>100 (N = 2)
>100 (N = 2)
>100 (N = 2)
>100 (N = 2)
>100 (N = 3)
>100 (N = 3)
-
OMe OMe
OMe
Br
CF3
F
-
H
H
H
F
F
-
H
H
H
H
H
-
68.59 ± 22.87 (N = 3)
67.3 ± 42.23 (N = 3)
2g
2h
2i
-
-
61.12 ± 4.31 (N = 2) 45.23 ± 24.67 (N = 3)
-
H
>100 (N = 2)
_ (N = 2)
35.37 ± 6.5 (N = 3)
28.61 ± 11.22 (N = 3)
>100 (N = 2)
2j
-
H
2k
2l
2-thiophenyl
-
-
>100 (N = 2)
-
H
-
F
-
H
H
-
85.23 ± 0.92 (N = 2) 56.07 ± 26.76 (N = 3)
2m
2n
2o
3-pyridyl
-
-
>100 (N = 2)
>100 (N = 2)
>100 (N = 2)
H
F
F
H
39.61 ± 12.88 (N = 3)
4

-
-
H
F
-
F
H
-
H
F
-
F
H
-
36
51
29
>100 (N = 2)
>100 (N = 2)
>100 (N = 2)
>100 (N = 2)
>100 (N = 2)
>100 (N = 2)
2p
2q
2r
5-
bromothiophenyl
-
H
OMe
H
H
21
>100 (N = 2)
>100 (N = 2)
2s
2.1.1.2
Synthesis of α-(3,5-dimethylphenoxy)-chalcone (2t – 2x)
3,5-dimethylphenol was reacted with 2ˊ-bromoacetophenone to synthesize 3,5-
dimethylphenoxy-1-phenylethanone. 3,5-dimethylphenoxy-1-phenylethanone was then
condensed with different aldehydes to prepare α-(3,5-dimethylphenoxy)-chalcone derivatives
(2t – 2x).
Table 1b: Structure, yields and inhibition of cell proliferation in HCC1954 and HCT116
cell lines.
Compound
R1
R2
R3
R4
R5
Yield
(%)
HCC195
HCT116
GI50 (µM) ± SD
GI50 (µM) ± SD
H
NO2
H
H
H
H
H
64
46
19.19 ± 0.17 (N = 2)
15.9 ± 1.57 (N = 2)
15.93 ± 4.39 (N = 2)
25.66 ± 0.01 (N = 2)
2t
OMe
OMe
OMe
2u
H
OMe
OMe
OMe
H
48
21.66 ± 3.59 (N = 2)
12.72 ± 2.28 (N = 2)
2v
Cl
OMe
H
H
H
Cl
H
45
44
54.95 ± 4.5 (N = 2)
11.94 ± 1.19 (N = 2)
50.88 ± 1.87 (N = 2)
17.99 ± 0.76 (N = 2)
2w
2x
H
OMe
OMe
2.1.1.1
Synthesis of α-(1,3-benzodioxol-5-yloxy)-chalcone (2aa – 2ae)
2-(benzo[d][1,3]dioxol-5-yloxy)-1-phenylethanone was prepared by reacting 2ˊ-
bromoacetophenone and sesamol. 2-(benzo[d][1,3]dioxol-5-yloxy)-1-phenylethanone was
then conensed with different aldehydes to prepare α-(1,3-benzodioxol-5-yloxy)-chalcone (2aa
– 2ae).
5

Table 1c: Structure, yields and inhibition of cell proliferation in HCC1954 and HCT116
cell lines.
Compound
R1
R2
R3
Yield
(%)
HCC195
HCT116
GI50 (µM) ± SD
46.97 ± 4.43 (N = 2)
41.02 ± 0.98 (N = 2)
61.36 ± 7.34 (N = 2)
>100 (N = 2)
GI50 (µM) ± SD
57.31 ± 1.43 (N = 2)
35.94 ± 3.39 (N = 2)
61.51 ± 16.54 (N = 2)
>100 (N = 2)
OMe
H
OMe
OMe
H
28
27
24
44
2aa
H
H
H
2ab
2ac
2ad
H
Br
H
NO2
H
H
Cl
13
>100 (N = 2)
>100 (N = 2)
2ae
2.1.2 Synthesis of α-COOEt chalcones (3a – 3e)
In order to prepare α-COOEt chalcones, Knoevenagel condensation was utilized: The ethyl
benzoacetate was condensed with different aldehydes using piperidine and acetic acid as a
catalyst in ethanol. Upon completion of the reaction, the product was purified by column
chromatography to afford pure α-COOEt chalcones.
Scheme 2.Synthetic routes of α-COOEt chalcones (3a- 3e). Reagents and conditions: (i).
Piperidine, EtOH, r.t., 48 hr.
Table 2: Structure, yields and inhibition of cell proliferation in HCC1954 and HCT116
cell lines.
6

Compound
R1
R2
R3
Yield
(%)
51
HCC1954
GI50 (µM) ) ± SD
14.3 ± 2.32
HCT116
GI50 (µM) ) ± SD
4.04 ± 0.04
N
H
H
H
H
OMe
Cl
F
2
2
2
2
2
3a
3b
3c
3d
3e
44
6.38 ± 1.86
6.64 ± 0.67
H
H
39
10.26 ± 1.79
8.27 ± 1.69
9.69 ± 0.95
H
OMe
H
H
44
6.04 ± 1.21
OMe
H
47
12.38 ± 1.12
9.65 ± 1.62
2.1.3 Synthesis of α-COOH chalcone (4)
2-benzoyl-3-(4-methoxyphenyl)acrylic acid was prepared through hydrolysis of 3e using
potassium hydroxide in ethanol. The product was portioned into basic aqueous layer, which
upon acidification by concentrated HCl yielded the precipitated product.
Scheme 3.Synthesis of 2-benzoyl-3-(4-methoxyphenyl)acrylic acid (4). Reagents and
conditions: (i). KOH, EtOH, r.t., 24 hr.
Table 3: Structure, yields and inhibition of cell proliferation in HCC1954 and HCT116
cell lines.
Yield (%)
HCC1954 GI50 (µM)
HCT116 GI50 (µM)
N
70
97.15 ± 1.15
72.7 ± 13.42
2
2.1.4 Synthesis of α-amide chalcones (5a – 5e)
7

Scheme 4: Synthesis of β-keto amide (i) (COCl)2, SOCl2, r.t., 24 hr.; (ii) Et3N, RNH2, r.t., 6
hr.
In order to prepare α-amide chalcones, compound 4 was converted to the corresponding acid
chloride using oxalyl chloride or thionyl chloride. The resulting acid chloride was then
reacted with different amines (aliphatic and aromatic) in the presence of trimethylamine in an
overnight reaction. The crude product was extracted in ethylacetate layer, given acidic
washings and purified using column chromatography to obtain the pure α-amide chalcones
(5a -5e).
Table 4: Structure, yields and inhibition of cell proliferation in HCC1954 and HCT116
cell lines.
Compounds
R
Yield (%)
HCC1954
GI50 (µM) ) ± SD
0.63 ± 0.06
HCT116
GI50 (µM) ) ± SD
0.69 ± 0.04
N
42
21
39
2
2
2
5a
5b
5c
7.53 ± 1.15
3.83 ± 0.83
21.88 ± 1.27
13.25 ± 0.28
28
35
6.61 ± 1.91
3.02 ± 0.17
1.59 ± 0.28
2
2
5d
5e
0.725 ± 0.05
2.2 Biology
2.2.1 Antiproliferative activities:
All the compounds were evaluated for their antiproliferative activity in colorectal (HCT116)
and breast (HCC1954) cancer cell lines. Half-maximal growth inhibition concentrations
(GI50) were determined in a 3-day Sulforhodamine (SRB) proliferation assay.
8

α-2-(naphthalen-2-yloxy)-1,3-diphenylprop-2-en-1-one (2a - 2s, Table 1a) were synthesized
and evaluated in HCC1954 and HCT116 cell lines. Compound 2c (R4 = Cl) showed better
activity (GI50 values of 19.35 µM and 23.54 µM in HCC1954 and HCT116 cell lines,
respectively) than other analogues. Nine compounds out of the 19 α-2-(naphthalen-2-yloxy)-
1,3-diphenylprop-2-en-1-one (2a-2s) were active with GI50 values ranging from 19 – 85 µM
in at least one or both of the cell lines. All these nine compounds contain halogen or a
trifluoromethyl group on the ring B. Compounds with nitro substitution (2a), mono and
trimethoxy pattern (2b, 2f, 2s), alkyl chain (2e) difluoro substitution (2p, 2q) and
heteroaromatic B ring (2l, 2r and 2n) were found inactive against HCC1954 and HCT116
cell lines (>100 M GI50). Five analogues of α-(3,5-dimethylphenoxy)-chalcone (with 4-Cl,
3-NO2, 2,3,4-trimethoxy, 4-Br) were synthesized and screened against HCC1954 and
HCT116 cell line with GI50 values ranging from 12 – 55 M (2t – 2x, Table 1b). Five
analogues of α-(1,3-benzodioxol-5-yloxy)-chalcones (2aa-2ae, Table 1c) differing in
substitution pattern on B ring (4-NO2, 4-Cl, 2,3,4-trimethoxy, 4-Br and unsubstituted phenyl
ring) were synthesized and evaluated for their anticancer activity in both the cell lines.
Maximum activity with GI50 values of 41.02 µM and 35.94 µM in HCC1594 and HCT116
cells respectively, was observed for compound 2ab having unsubstituted ring B while the
2,3,4-trimethoxy substitution pattern showed slightly higher GI50 values. Descending order of
anti-proliferative activity with respect to differentiation of substitution pattern on ring B is as
follows: unsubstituted B ring ˃ 2,3,4-trimethoxy ˃ 4-Br ˃ 4-Cl, 3-NO2.
Among α-COOEt chalcones (3a-3e, Table 2), compound 3a was found to be more active
against HCT116 cell line with a GI50 value of 4.04 µM (Table 2). α-COOEt chalcones 3a
and 3d with 4-MeO and 3-MeO groups on ring B, respectively, showed better anti-
proliferative activity than 2-MeO (3b) and 4-halogen substitutions on ring B (3c and 3e).
Compound 3a had the best activity and was, therefore, selected as a lead compound and used
for the preparation of further derivatives. It was first hydrolysed to yield α-COOH chalcone
(4) that had GI50 value of 97.15 µM and 72.7 µM in HCC1954 and HCT116, respectively.
The chalcone (4) was then reacted with various amines to prepare α-amide chalcones
(compounds 5a – 5e, Table 4). These compounds showed superior activity than α-COOEt, α-
COOH chalcones and α-aryloxy chalcones. Compounds 5a and 5e were found to be more
active in HCC1954 cell line with GI50 values of 0.63 µM and 0.725 µM, respectively. In
HCT116 cell line, compounds 5a and 5e exhibited GI50 values of 0.69 µM and 1.59 µM,
respectively. As the compounds 5a and 5e displayed the lowest GI50 values among the tested
compounds, these were chosen for further biological studies.
2.2.2 Compound 5a and 5e cause cell cycle arrest in S and G2/M phases
In order to determine the possible mechanism of action of these compounds, we determined
the cell cycle profile using FACS analysis in HCT116 cells following treatment with different
concentrations of both compounds for 24 hours. As shown in Figure 4, both compounds 5a
and 5e induced a G2/M cell cycle arrest at 6.25 µM and 12.5 µM concentrations compared
with the DMSO treated controls. Paclitaxel, a positive control completely arrested the cells in
the G2/M phase of the cell cycle. An increase in S phase of the cell cycle was also observed
following treatment of cells with both the compounds. These compounds are therefore,
capable of causing the cell cycle arrest in S and G2/M phase in HCT116 cell line.
9

Figure 4: Effect of 5a and 5e on cell cycle in HCT116 cells. A. HCT116 cells treated with
6.25M and 12.5M concentrations of 5a and 5e for 24 hours. Treatment with paclitaxel was
used as a control for G2/M cell cycle arrest. Cells were fixed, stained with propidium iodide
and analysed through FACS. B. Quantification of cells in different phases of the cell cycle
represented as relative percentage in each phase.
2.2.3 Compound 5a and 5e induce apoptosis and increase p53 levels in HCT116 cells.
We next determined the ability of compounds 5a and 5e to induced apoptosis in HCT116
cells. Cells were treated with three increasing concentrations (1.56, 3.13 and 6.25 µM) of
compound 5a or 5e for 24 hours and apoptosis was determined through western blotting
using antibodies specific for cleaved-PARP, an apoptotic marker. -actin was used as a
loading control. As shown in Figure 4, both compounds induced a robust, dose-dependent
increase in cleaved-PARP following 24-hour treatment. The increase in cleaved-PARP was
also accompanied by an increase in the expression of the pro-apoptotic p53 protein (Figure
5).
10

Figure 4: Compounds 5a and 5e induce apoptotic cell death in HCT116 cells. Cells treated
with three different concentrations of 5a and 5e for 24 hours were analysed by western
blotting using antibodies specific for Cleaved-PARP, p53 and β-actin.
2.2.4 p53 stabilization by 5a is not through post-translational mechanism:
We have recently shown that chalcone derivatives, SSE14105 and SSE14106, can cause
rapid accumulation of p53 in HCT116 cells through a post-translational mechanism.27 In
order to determine whether the increase in p53 levels following treatment with 5a and 5e in
HCT116 cells is regulated at the transcriptional or post-translational level, we measured p53
levels following treatment with 6.25 µM 5a in the presence or absence of cycloheximide
(CHX; an inhibitor of protein translation) for different time points. Treatment with 5a did not
have any effect on the stability of p53 protein following incubation with CHX at different
time points (Figure 5), indicating that 5a does not regulate p53 protein at the post-
translational level. The accumulation of p53 following 5a treatment, therefore, seems to be
regulated through a transcriptional mechanism.
Figure 5: p53 stabilization by compound 5a is not regulated at the post-translational level.
HCT116 cells were treated with DMSO or 6.25 µM 5a in the presence of CHX and samples
11

were taken after 10, 20, 30, 60 and 90 minutes of incubation and p53 levels were determined
through western blotting.
2.2.5 Compound 5a and 5e overcome multidrug resistance
Compounds 5a and 5e were next evaluated for their ability to overcome multidrug resistance
in two pairs of parental and MDR1/Pgp expressing cell lines (A2780/A2780-Pac-Res and
KB-3-1/KB-V1). As shown in Table 5, both the MDR1 expressing cell lines were highly
resistant to paclitaxel when compared to the parental controls (resistance index of 494 and
>1000 for A2780-Pac-Res and KB-V1, respectively). Compounds 5a and 5e however, were
able to overcome the multidrug resistance in both these cell lines with GI50 values
comparable to parental controls (Resistance index values of 1.15 and 1.28 for A2780-Pac-Res
and KB-V1, respectively).
Cell Lines
GI50 value
5e (μM)
5a (μM)
2.11±0.01 (2)
2.43±0. 07 (2)
1.15
Paclitaxel(nM)
0.433
A2780
A2780-Pac-Res
Resistance Index
KB-3-1
2.27±0.05 (2)
2.91±0.09 (2)
1.28
214
494
3.24±0.02 (2)
3.9±0.28 (2)
1.2
4.79±0.05 (2)
6.38±0.28 (2)
1.33
0.2085
>2000
>1000
KB-V1
Resistance Index
Table 5:
GI50 values of 5a, 5e and paclitaxel in two pairs of parental and MDR cell lines
(A2780/A2780-Pac-Res; Human ovarian cancer cell lines and KB-3-1/KB-V1;
cervical cancer cell lines)
We next determined whether 5a compound could directly inhibit the activity of Pgp in an in
vitro Pgp-Glo assay. As shown in Figure 6, there was no significant effect of 5a on the
ATPase activity of Pgp. Verapamil, a substrate of Pgp, on the other hand, enhanced its
ATPase activity by ~ 3 fold. This confirms that 5a overcomes multidrug resistance in these
cells by not being a substrate of Pgp.
12

Figure 6: Compound 5a does not inhibit Pgp ATPase activity in vitro. ATPase activity of
Pgp was measured using Pgp-Glo assay (Promega) from untreated, 100M Na3VO4,
200M Verapamil- and 10M 5a compound-treated Pgp reactions. Basal, verapamil-
treated and 5a-treated activity of the Pgp ATPase was calculated according to the
manufacturer’s instructions. ΔRLU= Difference in relative light units.
3. Conclusion
In the current work, we have presented the synthesis of forty chalcone-derived compounds
including thirty-seven novel compounds. All the compounds were evaluated for their
antiproliferative activities in colorectal (HCT116) and breast (HCC1954) cancer cell lines.
Two α-amide chalcones (5a and 5e) showed GI50 in the nanomolar range and were identified
as the most active compounds. Both these compounds were able to induce G2/M cell cycle
arrest and apoptotic cell death in HCT116 cells. Similarly, both the compounds increased p53
levels following 24-hour treatment. Finally, we evaluated these compounds (5a and 5e) for
their ability to overcome multidrug resistance in the MDR1 overexpressing multidrug
resistant cell lines (A2780-Pac-Res and KB-V1). Both compounds showed that they were
capable of overcoming the multidrug resistance without much loss in the activity. We also
determined that the 5a compound does not directly inhibit the ATPase activity of Pgp
(MDR1), in vitro, and hence overcomes multidrug resistance by being not a Pgp substrate.
Compounds 5a and 5e therefore could serve as novel hits for further lead optimization studies
to discover anticancer compounds with the ability to overcome multidrug resistance.
Conflict of interest
The authors declare that there is no conflict of interest.
Acknowledgement
The authors acknowledge the support by LUMS in the form of faculty initiative funds to
enable this research work.
13

4. Experimental
General Method:
Commercial reagents were used without further purification. Reagent grade solvents were
distilled before use. Silica gel 60 (230-400 mesh) from Merck was used for column
chromatography. Bruker, P Alpha Spectrometer was used for recording IR spectra. Proton
and carbon NMR spectra were recorded on Bruker Ascend 600 MHz. Thermo Scientific,
1300 GC-MS), was used to obtain GC-MS spectra. Melting points were taken on STUART
SMP3 melting point apparatus.
4.1 General procedure for the synthesis of α-aryloxy chalcones (2a – 2x, 2aa – 2ae)
Preparation of 2- Naphthol, 3,5-dimethylphenol and sesamol ethers (2´, 2´´, 2´´´): Phenol
(2-Naphthol, 3,5-dimethylphenol or sesamol) (1 equiv), 2´-bromoacetophenone (1 equiv)
were added to a reaction flask followed by addition of acetonitrile (10 mL). K2CO3 (2 equiv)
was added to the reaction flask and the reaction mixture was stirred at room temperature for
48 hours and the progress of the reaction was monitored using thin layer chromatography.45
The reaction mixture was then acidified with conc. HCl to bring pH to neutral. The crude
product was extracted into ethyl acetate (EtOAc) and purified through column
chromatography (using hexanes and ethyl acetate, 9:1). The 2-aryloxy-1-phenylethanones (2ˊ,
2ˊˊ, 2ˊˊˊ) were obtained in 90 -98% yields.
2-(naphthalen-2-yloxy)-1-phenylethanone (2ˊ): Brown solid (92.7%); m.p. 73-75 °C (Lit:
77 °C)45; 1H NMR (600 MHz, CDCl3) δ: 8.05 (d, J = 9.7 Hz, 2H), 7.78 (d, J = 9.1 Hz, 2H),
7.71 (d, J = 9.3 Hz, 1H), 7.63 (t, J = 7.4 Hz, 1H), 7.52 (t, J = 7.8 Hz, 2H), 7.44 (t, J = 8.1 Hz,
1H), 7.36 (t, J = 8.1 Hz, 1H), 7.28 (dd, J = 8.9, 2.6 Hz, 1H), 7.14 (d, J = 2.0 Hz, 1H), 5.38 (s,
2H); 13C NMR (151 MHz, CDCl3) δ 194.5 (C), 156.1 (C), 134.7 (C), 134.4 (C), 134.0 (CH),
129.9 (C), 129.5 (CH), 129.0 (CH), 128.3 (CH), 127.8 (CH), 127.0 (CH), 126.6 (CH), 124.2
(CH), 118.8 (CH), 107.4 (CH), 70.9 (CH2); MS m/z [M+] 264.12; FT-IR (ATR): 3050, 1699,
1628, 1597, 1211 cm-1.
2-(3,5-dimethylphenoxy)-1-phenylethanone (2ˊˊ): Brown solid (89.6%); m.p. 99.8-103 °C;
1H NMR (600 MHz, CDCl3)  ppm 8.02 (d, J = 9.7 Hz, 2H), 7.62 (t, J = 7.4 Hz, 1H), 7.51 (t,
13
J = 7.8 Hz, 2H), 6.65 (s, 1H), 6.59 (s, 2H), 5.24 (s, 2H), 2.29 (s, 6H); C NMR (151 MHz,
CDCl3)  ppm 194.8 (C), 158.2 (C), 139.5 (C), 134.8 (C), 133.9 (CH), 128.9 (CH), 128.3
(CH), 123.6 (CH), 112.6 (CH), 70.8 (CH2), 21.5 (CH3); MS m/z [M+] 240.12; FT-IR (ATR):
2916, 1705, 1593, 1510, 1462, 1417, 1323, 1246, 1211, 1164 cm-1.
2-(benzodioxol-5-yloxy)-1-phenylethanone (2ˊˊˊ): Brown solid (98.4%); m.p. 102-114 °C;
1H NMR (600 MHz, CDCl3)  ppm 7.97 (d, J = 9.8 Hz, 2H), 7.60 (t, J = 7.4 Hz, 1H), 7.48 (t,
J = 7.6 Hz, 2H), 6.67 (d, J = 8.5 Hz, 1H), 6.55 (d, J = 2.5 Hz, 1H), 6.34 (dd, J = 8.5, 2.6 Hz,
13
1H), 5.88 (s, 2H), 5.19 (s, 2H); C NMR (151 MHz, CDCl3)  ppm 194.7 (C), 153.6 (C),
148.4 (C), 142.4 (C), 134.6 (C), 133.9 (CH), 128.9 (CH), 128.1 (CH), 107.9 (CH), 106.1
(CH), 101.3 (CH2), 98.7 (CH), 71.8 (CH2); MS m/z [M+] 256.07; FT-IR (ATR): 1700, 1494,
1431, 1244, 1179, 1033 cm-1.
14

Synthesis of α-aryloxy chalcones (2a - 2x, 2aa – 2ae) from ethers: Compounds 2ˊ, 2ˊˊ and
2ˊˊˊ (1 equiv) were condensed with substituted aldehydes (1 equiv) in presence of KOH and
ethanol as solvent. The reaction mixture was stirred for 24 hours at room temperature. The
crude product was extracted into EtOAc layer and compounds (2a – 2x, 2aa – 2ae) were
purified through column chromatography (hexanes:ethyl acetate, 8:2).
2-(naphthalen-2-yloxy)-3-(4-nitrophenyl)-1-phenylprop-2-en-1-one (2a): White solid
(84.7%); m.p. 167.7 °C; 1H NMR (600 MHz, CDCl3)  ppm 8.05 (d, J = 8.9 Hz, 2H), 7.79 (d,
J = 8.8 Hz, 2H), 7.76 (d, J = 7.4 Hz, 2H), 7.57 (dd, J = 8.6, 3.5 Hz, 2H), 7.49 (d, J = 8.2 Hz,
1H), 7.36 (t, J = 7.5 Hz, 1H), 7.23-7.27 (m, 3H), 7.20 (t, J = 7.5 Hz, 1H), 7.12 – 7.09 (m,
1H), 7.05 (dd, J = 8.9, 2.5 Hz, 1H), 6.88 (s, 1H); 13C NMR (151 MHz, CDCl3)  ppm 191.7
(C), 153.5 (C), 151.0 (C), 147.7 (C), 139.5 (C), 136.1, 134.1 (C), 133.4 (C), 130.9 (CH),
130.5 (CH), 130.3 (C), 129.5 (CH), 128.6 (CH), 127.9 (CH), 127.2 (CH), 127.0 (CH), 125.1
(CH), 124.1 (CH), 123.5 (CH), 118.0 (CH), 111.5 (CH); MS m/z [M+] 395.12; FT-IR (ATR):
3057, 1663, 1625, 1462, 1164 cm-1.
3-(4-methoxyphenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2b): Viscous
1
liquid (47.3%); H NMR (600 MHz, CDCl3)  ppm 7.91 (d, J = 6.9 Hz, 2H), 7.77-7.71 (m,
4H), 7.65 (d, J = 9.5 Hz, 1H), 7.50 (t, J = 7.4 Hz, 1H), 7.45-7.37 (m, 3H), 7.33 (t, J = 8.1 Hz,
13
1H), 7.29-7.28 (m, 2H), 7.16 (s, 1H), 6.88 (d, J = 8.9 Hz, 2H), 3.80 (s, 3H); C NMR (151
MHz, CDCl3)  ppm 192.2 (C), 161.0 (C), 154.2 (C), 146.6 (C), 137.4 (CH), 134.3 (C), 132.5
(C), 132.5 (C), 130.2 (C), 130.0 (CH), 129.4 (CH), 129.3 (CH), 128.4 (CH), 127.8 (CH),
127.1 (CH), 126.6 (CH), 125.6 (CH), 124.5 (CH), 118.1 (CH), 114.4 (CH), 110.4 (CH), 55.4
(CH3); m/z [M+]: 310.06; FT-IR (ATR): 3055, 2837, 1653, 1628, 1596, 1508, 1462, 1246,
1211, 1164, 1027 cm-1.
3-(4-chlorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2c): Pale Yellow
(39.8%); m.p. 101-104 °C; 1H NMR (600 MHz, CDCl3)  ppm 7.78-7.777 (m, 2H), 7.60-7.59
(m, 4H), 7.51 (d, J = 8.3 Hz, 1H), 7.37 (t, J = 7.4 Hz, 1H), 7.30-7.24 (m, 3H), 7.22 – 7.19 (m,
13
3H), 7.13-7.10 (m, 2H), 6.93 (s, 1H); C NMR (151 MHz, CDCl3)  ppm 192.0 (C), 153.9
(C), 148.6 (C), 136.8 (C), 135.6 (C), 134.2 (C), 132.9 (CH), 131.7 (CH), 131.5 (C), 130.3
(CH), 130.1 (C), 129.4 (CH), 129.2 (CH), 128.5 (CH), 127.8 (CH), 127.2 (CH), 126.8 (CH),
126.7 CH), 124.8 (CH), 118.0 (CH), 110.8 (CH); MS m/z [M+] 384.09; FT-IR (ATR): 3057,
1657, 1627, 1597, 1510, 1489, 1463, 1357, 1246, 1212, 1165, 1131 cm-1.
3-(2-chlorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2d): White solid
(55.1%); m.p. 76.81 °C; 1H NMR (600 MHz, CDCl3)  ppm 7.95 (d, J = 9.6 Hz, 1H), 7.79 (d,
J = 6.9 Hz, 2H), 7.56-7.55 (m, 2H), 7.49 (d, J = 8.2 Hz, 1H), 7.36 (t, J = 7.5 Hz, 1H), 7.32 (s,
1H), 7.28-7.23 (m, 4H), 7.17 (t, J = 8.2 Hz, 1H), 7.12 (d, J= 2.3 Hz, 1H), 7.10-7.03 (m, 3H);
13C NMR (151 MHz, CDCl3)  ppm 191.9 (C), 154.1 (C), 149.1 (C), 136.7 (C), 134.9 (C),
134.2 (C), 133.0 (CH), 131.1 (C), 131.0 (CH), 130.5 (CH), 130.3 (CH), 130.1 (C), 129.9
(CH), 129.6 (CH), 128.5 (CH), 127.8 (CH), 127.2 (CH), 127.1 (CH), 126.8 (CH), 124.8
(CH), 123.5 (CH), 118.2 (CH), 111.0 (CH); MS m/z [M+] 384.09; FT-IR (ATR): 3057, 1659,
1596, 1509, 1463, 1438, 1243, 1210, 1162, 1124, 1051 cm-1.
15

3-(4-isopropylphenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2e): Viscous
1
liquid (74.2%); H NMR (600 MHz, CDCl3)  ppm 7.79 (d, J = 6.9 Hz, 2H), 7.63-7.61 (m,
4H), 7.53 (d, J = 9.6 Hz, 1H), 7.37 (t, J = 7.4 Hz, 1H), 7.30 – 7.26 (m, 3H), 7.21 (t, J = 8.1
Hz, 1H), 7.19 – 7.16 (m, 2H), 7.11 (d, J = 8.4 Hz, 2H), 7.04 (s, 1H), 2.77 (hept, J = 6.9 Hz,
1H), 1.12 (d, J = 7.0 Hz, 6H); 13C NMR (151 MHz, CDCl3)  ppm 192.2 (C), 154.2 (C),
151.2 (C), 147.6 (C), 137.2 (C), 134.3 (C), 132.6 (CH), 130.8 (CH), 130.5 (C), 130.2 (CH),
130.0 (C), 129.4 (CH), 129.0 (CH), 128.4 (CH), 127.8 (CH), 127.1 (CH), 127.1 (CH), 126.7
(CH), 124.5 (CH), 118.1 (CH), 110.5 (CH), 34.2 (CH), 23.8 (CH3); MS m/z [M+] 392.18; FT-
IR (ATR): 3054, 2960, 1656, 1597, 1510, 1462, 1316, 1245, 1210, 1164, 1130 cm-1.
2-(naphthalen-2-yloxy)-1-phenyl-3-(2,3,4-trimethoxyphenyl)prop-2-en-1-one
(2f):
1
Yellow solid (47.3%); m.p. 118-119.1 °C; H NMR (600 MHz, CDCl3)  ppm 7.94 – 7.89
(m, 3H), 7.74 (dd, J = 9.1, 6.0 Hz, 2H), 7.66 (d, J = 9.6 Hz, 1H), 7.55 (s, 1H), 7.51 (t, J = 7.4
Hz, 1H), 7.46 – 7.37 (m, 3H), 7.33 (t, J = 8.1 Hz, 1H), 7.30- 7.27 (m, 2H), 6.63 (d, J = 9.0
13
Hz, 1H), 3.94 (s, 3H), 3.88 (s, 3H), 3.84 (s, 3H); C NMR (151 MHz, CDCl3)  ppm 192.1
(C), 155.3 (C), 154.3 (C), 153.3 (C), 147.2 (C), 142.1 (C), 137.5 (C), 134.3 (C), 132.5 (CH),
130.1 (CH), 130.0 (C), 129.4 (C), 128.3 (C), 127.8 (CH), 127.1 (CH), 126.6 (CH), 125.8
(CH), 124.4 (CH), 123.3 (CH), 119.8 (C), 118.1 (CH), 110.4 (CH), 107.8 (CH), 61.8 (CH3),
61.0 (CH3), 56.1 (CH3); MS m/z [M+] 440.16; FT-IR (ATR): 3057, 2937, 1653, 1589, 1494,
1461, 1412, 1298, 1248, 1211, 1165, 1130 cm-1 .
3-(4-bromophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2g): Brown solid
(34.1%); m.p. 81.3-86 °C; 1H NMR (600 MHz, CDCl3)  ppm 8.13 (d, J = 6.3 Hz, 2H), 7.95
(dd, J = 8.5, 3.3 Hz, 2H), 7.89-7.86 (m, 3H), 7.74 – 7.70 (m, 3H), 7.64-7.61 (m, 3H), 7.56 (t,
J = 8.1 Hz, 1H), 7.49-7.49 (m, 2H), 7.45 (dd, J = 8.9, 2.5 Hz, 1H); 13C NMR (151 MHz,
CDCl3)  ppm 192.0 (C), 153.8 (C), 148.7 (C), 136.7 (C), 134.2 (C), 132.9 (CH), 132.2 (CH),
131.9 (CH), 131.9 (C), 130.3 (CH), 130.1 (C), 129.4 (CH), 128.5 (CH), 127.8 (CH), 127.2
(CH), 126.8 (CH), 126.7 (CH), 124.8 (CH), 124.0 (C), 118.0 (CH), 110.9 (CH); MS m/z [M+]
428.04; FT-IR (ATR): 3055, 1657, 1626, 1596, 1509, 1485, 1462, 1399, 1355, 1309, 1244,
1210, 1163, 1130, 1072, 1009 cm-1.
2-(naphthalen-2-yloxy)-1-phenyl-3-(4-(trifluoromethyl)phenyl)prop-2-en-1-one
(2h):
Viscous liquid (18.1%); 1H NMR (600 MHz, CDCl3)  ppm 7.81-7.78 (m, 4H), 7.61 (d, J =
8.6 Hz, 2H), 7.53 (d, J = 8.3 Hz, 1H), 7.50 (d, J = 8.3 Hz, 2H), 7.39 (t, J = 7.6 Hz, 1H), 7.29
13
(m, 3H), 7.23 (t, J = 7.6 Hz, 1H), 7.15 (s, 1H), 7.11 (d, J = 8.8 Hz, 1H), 6.96 (s, 1H); C
NMR (151 MHz, CDCl3)  ppm 191.9 (C), 153.7 (C), 149.8 (C), 136.4 (C), 136.4 (C), 134.1
(C), 133.2 (CH), 130.8 (q, J = 32 Hz, C), 130.5 (CH), 130.4 (CH), 130.2 (C), 129.5 (CH),
128.6 (CH), 127.8 (CH), 127.2 (CH), 126.9 (CH), 125.8 (q, J = 3 Hz, CH), 125.4 (CH), 124.9
(CH), 124.1 (q, J = 252.13 Hz, C), 118.0 (CH), 111.1 (CH); MS m/z [M+] 418.12; FT-IR
(ATR): 3057, 1662, 1598, 1511, 1463, 1320, 1246, 1212, 1165, 1123, 1067, 1016 cm-1.
3-(4-fluorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2i): Viscous liquid
(13.6%); 1H NMR (600 MHz, CDCl3)  ppm 7.76 (d, J = 7.0 Hz, 2H), 7.65 (dd, J = 8.8, 5.6
Hz, 2H), 7.58 (d, J = 8.6 Hz, 2H), 7.50 (d, J = 8.3 Hz, 1H), 7.36 (t, J = 7.5 Hz, 1H), 7.27-7.25
(m, 3H), 7.19 (t, J = 8.1 Hz, 1H), 7.13-7.10 (m, 2H), 6.95 (s, 1H), 6.90 (t, J = 8.7 Hz, 2H);
16

13C NMR (151 MHz, CDCl3)  ppm 192.1 (C), 163.4 (d, J = 252 Hz, C), 153.9 (C), 147.9
(C), 136.9 (C), 134.2 (C), 132.8 (CH), 132.6 (d, J = 8 Hz, CH), 130.3 (CH), 130.1 (C), 129.4
(CH), 129.2 (d, J = 3Hz, C), 128.5 (CH), 127.8 (CH), 127.2 (CH), 127.1 (CH), 126.8 (CH),
124.7 (CH), 118.0 (CH), 116.1 (CH), 116.0 (d, J = 21.7, CH), 110.7 (CH); MS m/z [M+]
368.12; FT-IR (ATR): 3057, 1656, 1597, 1505, 1463, 1211, 1157, 1010 cm-1.
3-(2-fluorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2j): Brown solid
1
(23.4%y); m.p. 131.0 °C; H NMR (600 MHz, CDCl3)  ppm 8.04 (t, J = 8.6 Hz, 1H), 7.82
(d, J = 6.7 Hz, 2H), 7.63 (d, J = 8.6 Hz, 2H), 7.55 (d, J = 8.2 Hz, 1H), 7.41 (t, J = 7.5 Hz,
1H), 7.33-7.29 (m, 3H), 7.26-7.19 (m, 3H), 7.16-7.14 (m, 1H), 7.02 – 6.98 (m, 2H); 13C NMR
(151 MHz, CDCl3)  ppm 191.8 (C), 160.9 (d, J = 252 Hz, C), 154.0 (C), 149.2 (C), 136.7
(C), 134.2 (C), 133.0 (CH), 131.4 (d, J = 8 Hz, CH), 130.7 (d, J = 2 Hz, CH), 130.3 (C),
130.1 (CH), 129.5 (CH), 128.5 (CH), 127.8 (CH), 127.2 (CH), 126.8 (CH), 124.7 (CH),
124.6 (d, J = 3 Hz, CH), 121. 1 (d, J = 9 Hz, C), 119.0 (d, J = 7 Hz, CH), 118.1 (CH), 115.6
(d, J = 22 Hz, CH), 111.0 (CH); MS m/z [M+] 368.12; FT-IR (ATR): 1651, 1625, 1231,
1194, 1116, 1032 cm-1.
3-(2, 5-difluorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2k): Light
1
brown solid (41.9%); m.p. 127.9 °C; H NMR (600 MHz, CDCl3)  ppm 7.94-7.90 (m, 3H),
7.75 (d, J = 9.1 Hz, 2H), 7.68 (d, J = 8.2 Hz, 1H), 7.54 (t, J = 7.5 Hz, 1H), 7.45-7.42 (m, 3H),
13
7.37 (t, J = 7.5 Hz, 1H), 7.30-7.25 (m, 3H), 7.11-7.07 (m, 1H), 7.05 – 7.01 (m, 1H); C
NMR (151 MHz, CDCl3)  ppm 191.6 (C), 158.6 (d, J = 256 Hz, C), 157.0 (d, J = 241 Hz,
C), 153.6 (C), 150.2 (C), 136.4 (C), 134.1 (C), 133.2 (CH), 130.4 (CH), 130.3 (C), 129.5
(CH), 128.6 (CH), 127.9 (CH), 127.2 (CH), 126.9 (CH), 124.9 (CH), 122.3 (dd, J = 13, 9 Hz,
C), 118.1 (CH), 117.7 (dd, J = 24, 8 Hz, CH), 117.1 (dd, J = 8, 2 Hz, CH), 116.8 (d, J = 2 Hz,
CH), 116.5 (dd, J = 25, 8 Hz, CH), 111.3 (CH); MS m/z [M+] 386.11; FT-IR (ATR): 3060,
2925, 1663, 1628, 1598, 1510, 1484, 1464, 1357, 1264, 1246, 1212, 1167, 1131 cm-1.
2-(naphthalen-2-yloxy)-1-phenyl-3-(thiophen-2-yl)prop-2-en-1-one (2l): Light brown
1
solid (41.9%); m.p. 127.9 °C; H NMR (600 MHz, CDCl3)  ppm 7.75 (d, J = 8.3 Hz, 2H),
7.58 (t, J = 9.1 Hz, 2H), 7.48 (d, J = 8.2 Hz, 1H), 7.36-7.34 (m, 2H), 7.29-7.22 (m, 5H), 7.18-
13
7.15 (m, 2H), 7.11-7.10 (m, 1H), 6.92 (t, J = 9.9 Hz, 1H); C NMR (151 MHz, CDCl3) 
ppm 191.0 (C), 153.8 (C), 145.8 (C), 137.2 (C), 135.5 (C), 134.3 (C), 132.7 (CH), 132.4
(CH), 131.2 (CH), 130.1 (C), 130.1 (CH), 129.3 (CH), 128.5 (CH), 127.8 (CH), 127.3 (CH),
127.2 (CH), 126.7 (CH), 124.5 (CH), 123.7 (CH), 118.0 (CH), 110.2 (CH); MS m/z [M+]
356.09; FT-IR (ATR): 3057, 1651, 1629, 1598, 1510, 1462, 1333, 1267, 1212, 1164 cm-1.
3-(3-fluorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2m): Viscous liquid
1
(69.0%); H NMR (600 MHz, CDCl3)  ppm 7.86 (d, J = 8.1 Hz, 2H), 7.68 (d, J = 9.1 Hz,
2H), 7.60 (d, J = 8.2 Hz, 1H), 7.54 (d, J = 10.3 Hz, 1H), 7.46 (t, J = 8.2 Hz, 2H), 7.37-7.34
(m, 3H), 7.30-7.26 (m, 2H), 7.22 (d, J = 2.5 Hz, 1H), 7.20 (dd, J = 8.9, 2.5 Hz, 1H), 7.01 –
6.99 (m, 2H); 13C NMR (151 MHz, CDCl3)  ppm 192.0 (C), 162.9 (d, J = 245 Hz, C), 153.8
(C), 149.1 (C), 136.6 (C), 135.0 (C), 135.0 (C), 134.2 (C), 133.0 (CH), 130.3 (CH), 130.2 (d,
J = 15 Hz, CH), 129.5 (CH), 128.5 (CH), 127.8 (CH), 127.2 (CH), 126.8 (CH), 126.5 (d, J =
2 Hz, CH), 126.2 (d, J = 2 Hz, CH), 124.8 (CH), 118.1 (CH), 116.8 (d, J = 27 Hz, CH), 116.7
17

(d, J = 21 Hz, CH), 111.0 (CH); MS m/z [M+] 368.12; FT-IR (ATR): 3057, 1658, 1596,
1509, 1463, 1445, 1355, 1277, 1210 cm-1.
2-(naphthalen-2-yloxy)-1-phenyl-3-(pyridin-3-yl)prop-2-en-1-one (2n): White solid
(73.7%); m.p. 78 °C; 1H NMR (600 MHz, CDCl3)  ppm 8.26 (s, 1H), 7.90 (s, 1H), 7.56 (d,
J = 8.0 Hz, 1H), 7.25 (d, J = 7.3 Hz, 2H), 7.06 (d, J = 9.2 Hz, 2H), 6.98 (d, J = 8.2 Hz, 1H),
6.85 (t, J = 7.4 Hz, 1H), 6.76-6.73 (m, 3H), 6.68 (t, J = 7.7 Hz, 1H), 6.64-6.60 (m, 2H), 6.57
(dd, J = 8.9, 2.5 Hz, 1H), 6.38 (s, 1H); 13C NMR (151 MHz, CDCl3)  ppm 191.6 (C), 153.6
(C), 151.3 (CH), 150.1 (CH), 150.0 (C), 136.9 (C), 136.4 (CH), 134.1 (C), 133.2 (CH), 130.4
(C), 130.2 (CH), 129.5 (CH), 129.3 (C), 128.6 (CH), 127.8 (CH), 127.2 (CH), 126.9 (CH),
124.9 (CH), 124.0 (CH), 123.5 (CH), 118.0 (CH), 111.0 (CH); MS m/z [M+] 356.09; FT-IR
(ATR): 1660, 1627, 1597, 1510, 1355, 1332, 1246, 1212, 1165, 1132 cm-1.
3-(3,4-difluorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2o): Viscous
1
liquid (11.9%); H NMR (600 MHz, CDCl3)  ppm 7.93 (d, J = 7.6 Hz, 2H), 7.78-7.75 (m,
3H), 7.68 (d, J = 8.4 Hz, 1H), 7.54-7.49 (m, 2H), 7.44-7.42 (m, 3H), 7.37 (t, J = 7.5 Hz, 1H),
7.28-7.25 (m, 2H), 7.20-7.15 (m, 1H), 7.02 (s, 1H); 13C NMR (151 MHz, CDCl3)  ppm
191.9 (C), 153.7 (C), 151.5 (dd, J = 253, 12 Hz, C), 149.8 (dd, J = 248, 12 Hz, C), 148.8 (d, J
= 2 Hz, C), 136.5 (C), 134.1 (C), 133.0 (CH), 130.4 (C), 130.2 (CH), 130.1 (dd, J = 6, 4 Hz,
C), 129.4 (CH), 128.5 (CH), 127.8 (CH), 127.3 (q, J = 3 Hz, CH), 127.2 (CH), 126.9 (CH),
125.3 (CH), 124.9 (CH), 119.0 (d, J = 18 Hz, CH), 118.0 (CH), 117.7 (d, J = 17 Hz, CH),
111.0 (CH); MS m/z [M+] 386.11; FT-IR (ATR): 3057, 1660, 1597, 1510, 1264, 1211, 1111,
1083, 1014 cm-1.
3-(3,5-difluorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2p): Yellow
1
solid (36.5%); m.p. 138.9 °C; H NMR (600 MHz, CDCl3)  ppm 7.91 (d, J = 8.2 Hz, 2H),
7.73 (dd, J = 8.6, 3.1 Hz, 2H), 7.66 (d, J = 8.3 Hz, 1H), 7.51 (t, J = 7.4 Hz, 1H), 7.43-7.39
(m, 3H), 7.37-7.35 (m, 3H), 7.28 (d, J = 2.6 Hz, 1H), 7.24 (dd, J = 9.1, 2.6 Hz, 1H), 6.95 (s,
1H), 6.80 (t, J = 8.7 Hz, 1H); 13C NMR (151 MHz, CDCl3)  ppm 191.8 (C), 163.1 (dd, J =
248, 12 Hz, C), 153.5 (C), 150.0 (C), 136.3 (C), 135.9 (t, J = 10 Hz, C), 134.1 (C), 133.2
(CH), 130.4 (CH), 130.2 (C), 129.4 (CH), 128.5 (CH), 127.8 (CH), 127.2 (CH), 126.9 (CH),
124.9 (CH), 124.4 (t, J = 2.8 Hz, CH), 118.0 (CH), 113.0 (dd, J = 20, 5 Hz, CH), 111.2 (CH),
104.9 (t, J = 25 Hz, CH); MS m/z [M+] 386.39; FT-IR (ATR): 3059, 2927, 1662, 1618, 1587,
1446, 1344, 1262, 1211, 1166, 1119 cm-1.
3-(2,4-difluorophenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2q): White
1
solid (51.4%); m.p. 122.6 °C; H NMR (600 MHz, CDCl3)  ppm 8.17 – 8.13 (m, 1H), 7.90
(d, J = 8.1 Hz, 2H), 7.71 (d, J = 8.6 Hz, 2H), 7.64 (d, J = 9.1 Hz, 1H), 7.49 (t, J = 7.5 Hz,
1H), 7.40-7.38 (m, 3H), 7.32 (t, J = 8.1 Hz, 1H), 7.26 (d, J = 3.6 Hz, 2H), 7.22 (dd, J = 8.8,
2.4 Hz, 1H), 6.86-6.81 (m, 2H); 13C NMR (151 MHz, CDCl3)  ppm 191.7 (C), 163.6 (dd, J
= 253, 12 Hz, C), 161.3 (dd, J = 246, 12 Hz, C), 153.8 (C), 149.0 (C), 136.6 (C), 134.2 (C),
133.0 (CH), 131.8 (dd, J = 9, 3 Hz, CH), 130.3 (CH), 130.2 (C), 129.5 (CH), 128.5 (CH),
127.8 (CH), 127.2 (CH), 126.9 (CH), 124.8 (CH), 118.0 (CH), 117.9 (dd, J = 6, 1 Hz, CH),
117.5 (dd, J = 11, 3 Hz, C), 112.1 (dd, J = 21, 3 Hz, CH), 111.0 (CH), 104.1 (t, J = 25 Hz,
CH); MS m/z [M+] 386.11; FT-IR (ATR): 3057, 1662, 1596, 1496, 1463, 1429, 1260, 1210,
1163, 1089 cm-1.
18

3-(5-bromothiophen-2-yl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2r): Black
1
purple solid (29.3%); m.p. 92.9 °C; H NMR (600 MHz, CDCl3)  ppm 7.92 (d, J = 8.0 Hz,
2H), 7.76 (m, 2H), 7.67 (d, J = 8.2 Hz, 1H), 7.51 (t, J = 7.4 Hz, 1H), 7.45-7.40 (m, 3H), 7.39
(s, 1H), 7.35 (t, J = 7.5 Hz, 1H), 7.32 (dd, J = 8.9, 2.5 Hz, 1H), 7.29 (d, J = 2.5 Hz, 1H), 7.11
(d, J = 4.0 Hz, 1H), 7.04 (d, J = 4.0 Hz, 1H); 13C NMR (151 MHz, CDCl3)  ppm 190.5 (C),
153.5 (C), 146.0 (C), 137.2 (C), 136.9 (C), 134.1 (C), 132.8 (CH), 132.2 (CH), 130.2 (C),
130.2 (CH), 130.1 (CH), 129.2 (CH), 128.5 (CH), 127.8 (CH), 127.2 (CH), 126.7 (CH),
124.7 (CH), 122.9 (CH), 119.1 (C), 117.9 (CH), 110.4 (CH); MS m/z [M+] 386.11; FT-IR
(ATR): 3056, 1660, 1597, 1510, 1462, 1417, 1323, 1246, 1211, 1164 cm-1.
3-(3-methoxyphenyl)-2-(naphthalen-2-yloxy)-1-phenylprop-2-en-1-one (2s): Viscous
liquid (21.6 %); 1H NMR (600 MHz, CDCl3)  ppm 7.77 (d, J = 9.7 Hz, 2H), 7.58 (d, J = 8.5
Hz, 2H), 7.50 (d, J = 9.5 Hz, 1H), 7.36 (t, J = 7.4 Hz, 1H), 7.27-7.23 (m, 4H), 7.21-7.17 (m,
2H), 7.14-7.11 (m, 2H), 7.11 (d, J = 2.2 Hz, 1H), 6.96 (s, 1H), 6.76 (dd, J = 8.3, 3.6 Hz, 1H),
3.58 (s, 3H); 13C NMR (151 MHz, CDCl3)  ppm 192.2 (C), 159.8 (C), 154.1 (C), 148.3 (C),
137.0 (C), 134.3 (C), 134.1 (C), 132.8 (CH), 130.2 (CH), 130.2 (C), 129.8 (CH), 129.5 (CH),
128.5 (CH), 128.4 (CH), 127.8 (CH), 127.2 (CH), 126.7 (CH), 124.6 (CH), 123.4 (CH),
118.0 (CH), 116.0 (CH),, 115.3 (CH), 110.6 (CH), 55.3 (CH3); MS m/z [M+] 380.14; FT-IR
(ATR): 3055, 2935, 1655, 1625, 1446, 1353, 1274, 1210, 1160, 1113, 1046 cm-1.
2-(3,5-dimethylphenoxy)-3-(3-nitrophenyl)-1-phenylprop-2-en-1-one (2t): Yellow solid
1
(64.4%); m.p. 135 - 136.6 °C; H NMR (600 MHz, CDCl3)  ppm 8.60 (s, 1H), 8.16 (d, J =
11.5 Hz, 1H), 8.10 (d, J = 7.9 Hz, 1H), 7.90 (d, J = 9.7 Hz, 2H), 7.57 – 7.52 (m, 2H), 7.44 (t,
13
J = 7.4 Hz, 2H), 6.93 (s, 1H), 6.61 (s, 1H), 6.60 (s, 2H), 2.20 (s, 6H); C NMR (151 MHz,
CDCl3)  ppm 191.8 (C), 155.6 (C), 150.5 (C), 148.6 (C), 139.9 (C), 136.4 (C), 135.7 (CH),
134.8 (C), 133.2 (CH), 129.8 (CH), 129.5 (CH), 128.6 (CH), 125.4 (CH), 124.9 (CH), 123.7
(CH), 123.5 (CH), 114.1 (CH), 21.4 (CH3); MS m/z [M+] 373.13; FT-IR (ATR): 3062, 2919,
1662, 1592, 1527, 1447, 1346, 1287, 1148, 1032 cm-1.
2-(3,5-dimethylphenoxy)-1-phenyl-3-(2,3,4-trimethoxyphenyl)prop-2-en-1-one (2u): Pale
1
yellow solid (46.9%); m.p. 107.4 – 113.4 °C; H NMR (600 MHz, CDCl3)  ppm 7.89-7.87
(m, 3H), 7.52 (t, J = 7.4 Hz, 1H), 7.43 (t, J = 7.4 Hz, 2H), 7.39 (s, 1H), 6.66 (d, J = 9.0 Hz,
13
1H), 6.63 (s, 2H), 6.61 (s, 1H), 3.89 (s, 3H), 3.86 (s, 3H), 3.86 (s, 3H), 2.22 (s, 6H); C
NMR (151 MHz, CDCl3)  ppm 192.1 (C), 156.4 (C), 155.2 (C), 153.1 (C), 147.3 (C), 142.1
(C), 139.6 (C), 137.7 (C), 132.3 (CH), 129.5 (CH), 128.3 (CH), 125.8 (CH), 124.6 (CH),
122.9 (CH), 120.0 (C), 113.7 (CH), 107.8 (CH), 61.8 (CH3), 61.0 (CH3), 56.1 (CH3), 21.4
(CH3); MS m/z [M+] 418.18; FT-IR (ATR): 2936, 1654, 1587, 1493, 1459, 1411, 1286, 1146,
1092, 1046 cm-1.
2-(3,5-dimethylphenoxy)-1-phenyl-3-(3,4,5-trimethoxyphenyl)prop-2-en-1-one
(2v):
Viscous liquid (48.8%); 1H NMR (600 MHz, CDCl3)  ppm 7.89 (d, J = 8.5 Hz, 2H), 7.53 (t,
J = 7.4 Hz, 1H), 7.44 (t, J = 7.7 Hz, 2H), 7.04 (s, 2H), 6.96 (s, 1H), 6.60 (s, 3H), 3.87 (s, 3H),
13
3.79 (s, 6H), 2.20 (s, 6H); C NMR (151 MHz, CDCl3)  ppm 192.2 (C), 156.2 (C), 153.2
(C), 147.6 (C), 139.7 (C), 139.6 (C), 137.3 (C), 132.6 (CH), 129.5 (CH), 128.6 (CH), 128.4
(CH) 128.4 (C), 124.7 (CH), 113.4 (CH), 108.0 (CH), 61.0 (CH3), 56.1 (CH3), 21.5 (CH3);
19

MS m/z [M+] 418.18; FT-IR (ATR): 1655, 1577, 1503, 1448, 1416, 1290, 1242, 1124, 1002
cm-1.
3-(2,6-dichlorophenyl)-2-(3,5-dimethylphenoxy)-1-phenylprop-2-en-1-one (2w): White
solid (45.7%); m.p. 117-118 °C; 1H NMR (600 MHz, CDCl3)  ppm 8.03 (d, J = 8.5 Hz, 2H),
7.57 (t, J = 7.5 Hz, 1H), 7.47 (t, J = 7.8 Hz, 2H), 7.28 (d, J = 8.1 Hz, 2H), 7.13 (t, J = 8.1 Hz,
1H), 6.79 (s, 1H), 6.56 (s, 2H), 6.52 (s, 1H), 2.15 (s, 6H); 13C NMR (151 MHz, CDCl3) 
ppm 191.3 (C), 155.9 (C), 151.2 (C), 139.2 (C), 136.6 (C), 135.0 (C), 133.2 (CH), 131.5 (C),
129.8 (CH), 129.5 (CH), 128.5 (CH), 127.8 (CH), 124.9 (CH), 119.8 (CH), 114.7 (CH), 21.3
(CH3); MS m/z [M+] 396.07; FT-IR (ATR): 1654, 1579, 1501, 1448, 1417, 1289, 1245, 1122,
1014 cm-1.
2-(3,5-dimethylphenoxy)-1-phenyl-3-(2,4,5-trimethoxyphenyl)prop-2-en-1-one
(2x):
White solid (44.6%); m.p. 128.13 °C; 1H NMR (600 MHz, CDCl3)  ppm 7.89 (d, J = 8.2 Hz,
2H), 7.72 (s, 1H), 7.58 (s, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.43 (t, J = 7.6 Hz, 2H), 6.61 (s, 2H),
13
6.58 (s, 1H), 6.47 (s, 1H), 3.90 (s, 3H), 3.81 (s, 3H), 3.66 (s, 3H), 2.20 (s, 6H); C NMR
(151 MHz, CDCl3)  ppm 192.1 (C), 156.5 (C), 153.8 (C), 151.7 (C), 145.9 (C), 143.0 (C),
139.5 (C), 137.9 (C), 132.1 (CH), 129.5 (CH), 128.2 (CH), 124.3 (CH), 123.5 (CH), 113.6
(C), 113.2 (CH), 113.1 (CH), 96.4 (CH), 56.6 (CH3), 56.1 (CH3), 56.0 (CH3), 21.4 (CH3); MS
m/z [M+] 418.18; FT-IR (ATR): 2922, 1664, 1603, 1509, 1464, 1408, 1269, 1207, 1122,
1024 cm-1.
2-(benzo[d][1,3]dioxol-5-yloxy)-1-phenyl-3-(2,3,4-trimethoxyphenyl)prop-2-en-1-one
(2aa): White solid (28.5%); m.p. 145.6 °C; 1H NMR (600 MHz, CDCl3) δ ppm 7.88-7.85 (m,
, 3H), 7.50 (t, J = 7.4 Hz, 1H), 7.41 (t, J = 7.7 Hz, 2H), 7.37 (s, 1H), 6.66 (d, J = 9.0 Hz, 1H),
6.60 (d, J = 8.4 Hz, 1H), 6.58 (s, 1H), 6.43 (dd, J = 8.5, 2.5 Hz, 1H), 5.83 (s, 2H), 3.87 (s,
13
3H), 3.85 (s, 3H), 3.84 (s, 3H); C NMR (151 MHz, CDCl3)  ppm 191.9 (C), 155.2 (C),
153.0 (C), 151.4 (C), 148.4 (C), 147.4 (C), 143.1 (C), 142.0 (C), 137.5 (C), 132.4 (CH),
129.3 (CH), 128.2 (CH), 125.6 (CH), 122.6 (CH), 119.7 (C), 108.0 (CH), 107.7 (CH), 107.5
(CH), 101.4 (CH2), 99.0 (CH), 61.7 (CH3), 60.9 (CH3), 56.0 (CH3).MS m/z [M+] 434.14; FT-
IR (ATR): 3057, 2939, 1653, 1589, 1480, 1412 1244, 1171, 1137, 1092, 1037 cm-1.
2-(benzo[d][1,3]dioxol-5-yloxy)-1,3-diphenylprop-2-en-1-one (2ab): Viscous liquid
1
(27.5%); H NMR (600 MHz, CDCl3)  ppm 7.90 (d, J = 7.0 Hz, 2H), 7.79 (d, J = 9.8 Hz,
2H), 7.54 (t, J = 7.4 Hz, 1H), 7.44 (t, J = 7.8 Hz, 2H), 7.40 – 7.36 (m, 3H), 6.98 (s, 1H), 6.63-
13
6.61 (m, 2H), 6.46 (dd, J = 8.4, 2.6 Hz, 1H), 5.85 (s, 2H); C NMR (151 MHz, CDCl3) 
ppm 192.1 (C), 151.1 (C), 148.5 (C), 148.4 (C) 143.3 (C), 137.0 (C), 132.9 (C), 132.7 (CH),
130.4 (CH), 129.7 (CH), 129.3 (CH), 128.8 (CH), 128.4 (CH), 127.7 (CH), 108.1 (CH),
107.8 (CH), 101.4 (CH2), 99.3 (CH); MS m/z [M+] 344.10; FT-IR (ATR): 3056, 2891, 1656,
1479, 1447, 1244, 1177, 1138, 1036 cm-1.
2-(benzo[d][1,3]dioxol-5-yloxy)-3-(4-bromophenyl)-1-phenylprop-2-en-1-one
(2ac):
Viscous liquid (24.2%); 1H NMR (600 MHz, CDCl3)  ppm 7.85 (d, J = 8.5 Hz, 2H), 7.62 (d,
J = 8.6 Hz, 2H), 7.54 (t, J = 7.4 Hz, 1H), 7.49 (d, J = 8.6 Hz, 2H), 7.43 (t, J = 7.7 Hz, 2H),
6.85 (s, 1H), 6.59 (d, J = 8.4 Hz, 1H), 6.53 (d, J = 2.5 Hz 1H), 6.38 (dd, J = 8.4, 2.5 Hz, 1H),
5.88 (s, 2H); 13C NMR (151 MHz, CDCl3)  ppm 192.1 (C), 150.9 (C), 149.1 (C), 148.6 (C),
20

143.6 (C), 136.9 (C), 133.0 (CH), 132.1 (CH), 131.9 (C), 131.9 (CH), 129.4 (CH), 128.5
(CH), 126.0 (CH), 123.9 (C), 108.2 (CH), 108.1 (CH), 101.6 (CH2), 99.5 (CH); MS m/z [M+]
422.02; FT-IR (ATR): 3056, 2921, 2854, 2776, 1659, 1478, 1399, 1245, 1171, 1139, 1073,
1036, 1009 cm-1.
2-(benzo[d][1,3]dioxol-5-yloxy)-3-(3-nitrophenyl)-1-phenylprop-2-en-1-one (2ad): Light
brown solid (43.5%); m.p. 95.2 °C; 1H NMR (600 MHz, CDCl3)  ppm 8.60 (s, 1H), 8.17 (m,
1H), 8.07 (d, J = 7.9 Hz, 1H), 8.08 (d, J = 7.9 Hz, 1H), 7.88 (d, J = 9.6 Hz, 2H), 7.57-7.53
(m, 2H), 7.44 (t, J = 7.8 Hz, 2H), 6.88 (s, 1H), 6.59 (d, J = 8.4 Hz, 1H), 6.54 (d, J = 2.5 Hz,
1H), 6.39 (dd, J = 8.4, 2.5 Hz, 1H), 5.87 (s, 2H); 13C NMR (151 MHz, CDCl3)  ppm 191.7
(C), 150.6 (C), 150.6 (C), 148.7 (C), 148.6 (C), 143.9 (C), 136.3 (C), 135.7 (CH), 134.7 (C),
133.3 (CH), 129.8 (CH), 129.4 (CH), 128.6 (CH), 124.8 (CH), 123.8 (CH), 123.1 (CH),
108.3 (CH), 108.2 (CH), 101.7 (CH2), 99.6 (CH); MS m/z [M+] 389.09; FT-IR (ATR): 2922,
1666, 1528, 1501, 1348, 1246, 1173, 1091, 1036 cm-1.
2-(benzo[d][1,3]dioxol-5-yloxy)-3-(4-chlorophenyl)-1-phenylprop-2-en-1-one
(2ae):
Viscous liquid (12.5%); 1H NMR (600 MHz, CDCl3)  ppm 7.86 (d, J = 9.7 Hz, 2H), 7.69 (d,
J = 8.6 Hz, 2H), 7.54 (t, J = 7.4 Hz, 1H), 7.43 (t, J = 7.8 Hz, 2H), 7.33 (d, J = 8.6 Hz, 2H),
6.88 (s, 1H), 6.60 (d, J = 8.4 Hz, 1H), 6.54 (d, J = 2.5 Hz, 1H), 6.40 (dd, J = 8.4, 2.5 Hz, 1H),
5.87 (s, 2H); 13C NMR (151 MHz, CDCl3)  ppm 192.0 (C), 151.0 (C), 148.9 (C), 148.6 (C),
143.5 (C), 136.9 (C), 135.5 (C), 132.9 (CH), 131.6 (CH), 131.5 (C), 129.4 (CH), 129.1 (CH),
128.5 (CH), 126.0 (CH), 108.2 (CH), 108.0 (CH) 101.6 (CH2), 99.4 (CH); MS m/z [M+]
378.08; FT-IR (ATR): 3055, 2892, 1656, 1479, 1245, 1171, 1138, 1089, 1036 cm-1.
4.1 Procedure for the synthesis of Ethyl 2-benzoyl-3-phenyl acrylate (3a-3e)
Ethyl benzoyl acetate (1 equiv) and the aldehyde (1 equiv) were dissolved in 5 mL ethanol in
a 50 mL round bottom flask. Piperidine (0.1 equiv) and acetic acid (0.5 equiv) were added to
the reaction flask and the reaction mixture was stirred at room temperature for 48 hours.
Crude product was dissolved in ethyl acetate 25 mL, transferred to separating funnel and
washed with 5% HCl solution. The compound was further purified through column
chromatography (hexanes: ethyl acetate).
Ethyl 2-benzoyl-3-(4-methoxyphenyl)acrylate (3a): White solid (51.1%); m.p. 91.3 °C
(Lit: 94 °C)46; 1H NMR (600 MHz, CDCl3)  ppm 7.96 (d, J = 9.4 Hz, 2H), 7.90 (s, 1H), 7.55
(t, J = 8.0 Hz, 1H), 7.43 (t, J = 7.8 Hz, 2H), 7.31 (d, J = 8.9 Hz, 2H), 6.74 (d, J = 8.9 Hz, 2H),
4.20 (q, J = 7.1 Hz, 2H), 3.74 (s, 3H), 1.15 (t, J = 7.0 Hz, 3H); 13C NMR (151 MHz, CDCl3)
 ppm 196.3 (C), 165.4 (C), 161.5 (C), 142.4 (CH), 136.4 (C), 133.9 (CH), 132.4 (CH), 129.3
(CH), 128.9 (CH), 128.7 (C), 125.6 (C), 114.4 (CH), 61.4 (CH2), 55.4 (CH3), 14.2 (CH3); m/z
[M+]: 310.06; FT-IR (ATR): 2936, 1713, 1668, 1598, 1510, 1243, 1169 cm-1.
1
Ethyl 2-benzoyl-3-(4-chlorophenyl)acrylate (3b): Viscous liquid (44.0 %); H NMR (600
MHz, CDCl3)  ppm 7.92 (d, J = 9.7 Hz, 2H), 7.89 (s, 1H), 7.56 (t, J = 7.4 Hz, 1H), 7.43 (t, J
= 7.8 Hz, 2H), 7.28 (d, J = 8.6 Hz, 2H), 7.20 (d, J = 8.6 Hz, 2H), 4.22 (q, J = 7.1 Hz, 2H),
13
1.16 (t, J = 7.1 Hz, 3H); C NMR (151 MHz, CDCl3)  ppm 195.4 (C), 164.8 (C), 141.1
(CH), 136.5 (C), 136.0 (C), 134.1 (CH), 132.0 (C), 131.3 (CH), 131.0 (C), 129.1 (CH), 128.9
21

(CH), 61.7 (CH2), 14.0 (CH3).; MS m/z [M+]: 314.07; FT-IR (ATR): 1716, 1670, 1621, 1590,
1490, 1246, 1227, 1192, 1082, 1012 cm-1.
1
Ethyl 2-benzoyl-3-(4-fluorophenyl)acrylate (3c): Viscous liquid (39.8%); H NMR (600
MHz, CDCl3)  ppm 7.93 (d, J = 8.0 Hz, 2H), 7.90 (s, 1H), 7.54 (t, J = 7.4 Hz, 1H), 7.41 (t, J
= 7.8 Hz, 2H), 7.34 (dd, J = 8.9, 5.3 Hz, 2H), 6.89 (t, J = 8.6 Hz, 2H), 4.20 (q, J = 7.1 Hz,
2H), 1.14 (t, J = 7.2 Hz, 3H); 13C NMR (151 MHz, CDCl3)  ppm 195.5 (C), 164.9 (C),
163.7 (d, J = 252.6 Hz, C), 141.2 (CH), 136.1 (C), 134.0 (CH), 132.3 (d, J = 8.7 Hz, CH),
131.1 (C), 129.2 (CH), 129.1 (C), 128.9 (CH), 116.0 (d, J = 21.9 Hz, CH), 61.6 (CH2), 14.0
(CH3); m/z [M+]: 298.10; FT-IR (ATR): 1717, 1670, 1598, 1508, 1224, 1191, 1161, 1085 cm-
1
.
Ethyl 2-benzoyl-3-(3-methoxyphenyl)acrylate (3d): Viscous liquid (44.0%); 1H NMR (600
MHz, CDCl3)  ppm 7.95 (d, J = 8.3 Hz, 2H), 7.92 (s, 1H), 7.55 (t, J = 7.4 Hz, 1H), 7.43 (t, J
= 7.5 Hz, 2H), 7.14 (t, J = 8.0 Hz, 1H), 6.95 (d, J = 7.7 Hz, 1H), 6.85 (m, 1H), 6.82 (dd, J =
13
8.3, 3.5 Hz, 1H), 4.23 (q, J = 7.1 Hz, 2H), 3.60 (s, 3H), 1.18 (t, J = 7.1 Hz, 3H); C NMR
(151 MHz, CDCl3)  ppm 195.7 (C), 165.1 (C), 159.7 (C), 142.6 (CH), 136.3 (C), 134.2
(CH), 134.0 (C), 131.6 (C), 129.9 (CH), 129.2 (CH), 129.0 (CH), 123.1 (CH), 117.0 (CH),
114.6 (CH), 61.7 (CH2), 55.2 (CH3), 14.2 (CH3); m/z [M+]: 314.07; FT-IR (ATR): 1717,
1671, 1578, 1224, 1190, 1171 cm-1.
1
Ethyl 2-benzoyl-3-(2-methoxyphenyl)acrylate (3e):Viscous liquid (47.0%); H NMR (600
MHz, CDCl3)  ppm 8.34 (s, 1H), 7.98 (d, J = 9.6 Hz, 2H), 7.55 (t, J = 7.4 Hz, 1H), 7.44 (t, J
= 7.7 Hz, 2H), 7.28 (d, J = 7.9 Hz, 2H), 6.84 (d, J = 8.2 Hz, 1H), 6.77 (t, J = 7.6 Hz, 1H),
4.26 (q, J = 7.1 Hz, 2H), 3.74 (s, 3H), 1.21 (t, J = 7.1 Hz, 3H); 13C NMR (151 MHz, CDCl3)
 ppm 195.3 (C), 165.3 (C), 157.8 (C), 138.5 (CH), 136.6 (C), 133.4 (CH), 131.9 (CH), 131.0
(C), 130.5 (CH), 129.0 (CH), 128.6 (CH), 122.2 (C), 120.5 (CH), 110.8 (CH), 61.3 (CH2),
55.1 (CH3), 14.0 (CH3); m/z [M+]: 314.07; FT-IR (ATR): 1715, 1670, 1596, 1463, 1258,
1224, 1191, 1115, 1084 cm-1.
4.1 Procedure for synthesis of 2-benzoyl-3-(4-methoxyphenyl)acrylic acid (4)
Compound 3a (1 equiv) was dissolved in 10mL ethanol, 40mL of water was added followed
by slow addition of KOH (15 equiv) into the reaction mixture. The mixture was stirred at
room temperature for 6 hours. Ethanol was removed using rotary evaporator. The reaction
mixture was dissolved into aqueous bicarbonate solution and washed with EtOAc to remove
unreacted ester. The acidification of the aqueous layer with HCl led to precipitation of the
product which was then filtered, washed with water and dried to obtain product as white solid
(70.0%); m.p. 180 °C; 1H NMR (600 MHz, CDCl3)  ppm 7.98 (s, 1H), 7.96 (d, J = 8.4 Hz,
2H), 7.55 (t, J = 7.4 Hz, 1H), 7.43 (t, J = 7.7 Hz, 2H), 7.31 (d, J = 8.9 Hz, 2H), 6.75 (d, J =
13
8.9 Hz, 2H), 3.75 (s, 3H); C NMR (151 MHz, CDCl3)  ppm 196.0 (C), 170.2 (C), 162.0
(C), 144.9 (CH), 136.1 (C), 134.2 (CH), 132.9 (CH), 129.4 (CH), 129.1 (CH), 127.3 (C),
125.3 (C), 114.5 (CH), 55.5 (CH3); m/z [M+]: 238.10; FT-IR (ATR): 2500-3500, 1671, 1598,
1512, 1260, 1175 cm-1.
4.1 General procedure for synthesis of α-substituted chalcones (5a - 5e)
22

Compound 4 (1 equiv) was stirred in SOCl2 (3 equiv) for 24 hours. Toluene was added in the
reaction mixture and excess thionyl chloride was removed through distillation using dropping
funnel. 5 mL dichloromethane, amine (1.2 equiv) and trimethylamine (4 equiv) were added in
reaction mixture and allowed to stir for 3 hours. Organic compound was extracted in ethyl
acetate layer and acidic washing was done to remove unreacted to remove triethylamine.
Desired product was purified through column chromatography (hexanes: ethyl acetate).
2-Benzoyl-N-benzyl-3-(4-methoxyphenyl)acrylamide (5a): White solid (42.7%); m.p.
130.7 °C; 1H NMR (500 MHz, DMSO-d6) δ ppm 8.78 (t, J = 6.0 Hz, 1H), 7.89 (d, J = 9.6 Hz,
2H), 7.64-7.61 (m, 2H), 7.50 (t, J = 9.0 Hz, 2H), 7.31 (d, J = 7.1 Hz, 2H), 7.27-7.23 (m, 5H),
6.84 (d, J = 8.8 Hz, 2H), 4.36 (d, J = 6.0 Hz, 1H), 3.69 (s, 1H); 13C NMR (126 MHz, DMSO-
d6)  ppm 196.8 (C), 165.1 (C), 160.2 (C), 139.4 (C), 136.1 (CH), 135.7 (C), 134.0 (C), 133.0
(CH), 131.2 (CH), 129.0 (CH), 128.9 (CH), 128.3 (CH), 127.2 (CH), 126.8 (C), 125.8 (CH),
114.3 (CH), 55.2 (CH3), 42.5 (CH2); MS m/z [M+]: 371.21; FT-IR (ATR): 3250, 1661, 1601,
1511, 1248, 1176 cm-1.
Ethyl 2-(2-benzoyl-3-(4-methoxyphenyl)acrylamido)acetate (5b): Viscous liquid (20.5%);
1H NMR (600 MHz, CDCl3)  ppm 7.97 (d, J = 9.7 Hz, 2H), 7.92 (s, 1H), 7.55 (t, J = 7.4 Hz,
1H), 7.43 (t, J = 7.7 Hz ,2H), 7.32 (d, J = 8.9 Hz, 2H), 6.75 (d, J = 8.9 Hz, 2H), 4.30-4.29
(m, 2H), 3.75 (s, 3H), 3.52 (t, J = 4.9 Hz, 2H), 3.32 (q, J = 7.0 Hz, 2H), 1.05 (t, J = 7.0 Hz,
3H); 13C NMR (151 MHz, CDCl3)  ppm 196.2 (C), 165.4 (C), 161.6 (C), 142.8 (CH), 136.5
(C), 133.9 (CH), 132.5 (CH), 129.4 (CH), 128.9 (CH), 128.4 (C), 125.5 (C), 114.4 (CH), 68.2
(CH2), 66.7 (CH2), 64.8 (CH2), 55.4 (CH3), 15.2 (CH3); MS m/z [M+] 381.10; FT-IR (ATR):
3307, 1729, 1688, 1639, 1395, 1179 cm-1.
2-Benzoyl-N-isopropyl-3-(4-methoxyphenyl)acrylamide (5c): White solid (38.9%); m.p.
101.1-102 °C; 1H NMR (500 MHz, DMSO-d6) δ ppm 8.10 (d, J = 7.7 Hz, 1H), 7.86 (dd, J =
8.4 Hz, 1.6 Hz, 2H), 7.61 (t, J = 7.4 Hz, 1H), 7.50-7.47 (m, 3H), 7.21 (d, J = 8.9 Hz, 2H),
6.83 (d, J = 8.9 Hz, 2H), 3.92 (m, 1H), 3.69 (s, 3H), 1.12 (d, J = 6.6 Hz, 6H); 13C NMR (126
MHz, DMSO-d6)  ppm 196.8 (C), 164.6 (C), 160.1 (C), 136.2 (C), 134.8 (C), 133.8 (CH),
133.6 (CH), 131.0 (CH), 128.9 (CH), 128.8 (CH), 125.9 (C), 114.2 (CH), 55.2 (CH3), 40.9
(CH), 22.2 (CH3); MS m/z [M+] 323.16; FT-IR (ATR): 3316, 1659, 1637, 1600, 1509, 1254,
1128 cm-1.
2-Benzoyl-3-(4-methoxyphenyl)-N-propylacrylamide (5d): White solid (28.0%); m.p.
114.7 °C; 1H NMR (600 MHz, DMSO-d6) δ ppm 8.18 (t, J = 5.7 Hz, 1H), 7.86 (d, J = 9.7 Hz,
2H), 7.60 (t, J = 7.4 Hz, 1H), 7.51 (s, 1H), 7.48 (t, J = 7.8 Hz, 2H), 7.21 (d, J = 8.9 Hz, 2H),
6.82 (d, J = 8.9 Hz, 2H), 3.68 (s, 3H), 3.09 (q, J = 6.6 Hz, 2H), 1.46 (h, J = 7.3 Hz, 2H), 0.83
(t, J = 7.4 Hz, 3H); 13C NMR (151 MHz, DMSO-d6)  ppm 196.8 (C), 165.0 (C), 160.1 (C),
136.1 (C), 135.1 (CH), 133.9 (CH), 133.4 (C), 131.0 (CH), 128.9 (CH), 128.8 (CH), 125.8
(C), 114.2 (CH), 55.2 (CH3), 40.8 (CH2), 22.2 (CH2), 11.4 (CH3); MS m/z [M+] 323.16; FT-
IR (ATR): 3325, 1661, 1599, 1504,1255, 1175, 1027 cm-1.
2-Benzoyl-N-(4-chlorobenzyl)-3-(4-methoxyphenyl)acrylamide (5e): Viscous liquid
(35.0%); 1H NMR (500 MHz, DMSO-d6) δ ppm 8.79 (t, J = 6.0 Hz, 1H), 7.89-7.87 (m, 2H),
7.64-7.62 (m, 2H), 7.50 (t, J = 8.2 Hz, 2H), 7.37 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.5 Hz, 2H),
23

7.23 (d, J = 8.8 Hz, 2H), 6.84 (d, J = 8.9 Hz, 2H), 4.33 (d, J = 6.0 Hz, 2H), 3.69 (s, 3H); 13C
NMR (126 MHz, DMSO-d6)  ppm 196.8 (C), 165.1 (C), 160.3 (C), 138.5 (C), 136.1 (C),
135.9 (C), 134.1 (C), 132.8 (CH), 131.3 (CH), 131.2 (CH), 129.1 (CH), 129.0 (CH), 128.9
(CH), 128.2 (CH), 125.7 (C), 114.3 (CH), 55.2 (CH3), 41.9 (CH2); MS m/z [M+] 405.11; FT-
IR (ATR): 3369, 3054, 1663, 1598, 1551, 1127, 1091 cm-1.
Cell culture and proliferation assays:
Cell lines were grown in their recommended culture media (DMEM or RPMI) supplemented
with 10% FBS and 1x antimycotic/antibiotic solution at 37°C in humidified incubators with
5% CO2. Antiproliferative activities of the compounds against HCC1954 and HCT116 cell
lines were determined using a 3-day sulforhodamine B (SRB) proliferation assay as described
previously (Safia et al). Cells were plated in 96 well plates and treated the next day with nine
3-fold dilutions of the compounds. Cells were incubated with the compounds for 72 hours at
37°C in humidified incubators with 5% CO2. At the end of treatment, cells were fixed with
3% ice-cold trichloroacetic acid (TCA) at 4°C for at least 2 hours. Plates were washed with
water, air-dried and stained with 0.06% SRB for 30 minutes at room temperature. Stained
cells were washed with 0.1% acetic acid, air dried and bound SRB bound to the cells was
dissolved in 10mM Tris pH 10.5. O.D. was measured at 490nm on a BioTek microplate
reader and GI50 values were calculated using GraphPad Prism.
Cell Cycle Analysis:
Cell cycle profile of HCT116 cells treated with compounds 5a and 5e was determined
through fluorescence-activated cell sorting (FACS) as described previously (Safia et al).
Briefly, cells were treated with indicated concentrations for 24 hours, fixed with 85% ethanol
and stained with propidium iodide/RNase solution (10g/ml PI and 0.5% RNase) at 37°C for
30 minutes. Fixed and stained samples were analysed using BD FACSCalibur.
Immunoblotting:
For PARP-cleavage and p53 induction, HCT116 cells were plated in 6-well plates and treated
with different concentrations of 5a and 5e for 24 hours. Following treatment, cells were lysed
with lysis buffer containing 50mM NaCl, 25mM Tris pH 7.5, 1% Triton-X 100 and
supplemented with phosphatase and protease inhibitors. The protein concentration of cell
lysates was measured using Bradford and equal amounts of proteins were separated on SDS-
PAGE. Proteins were transferred onto nitrocellulose membranes and incubated overnight at
4°C with primary antibodies specific for cleaved-PARP, p53 and  actin. Next day,
membranes were washed with PBS containing 0.1% Tween 20 (PBST) and incubated for 1
hour with HRP-labelled secondary antibodies. Membranes were washed again with PBST
solution, developed using ECL reagent and imaged on BioRad Chemidoc. For p53
stabilization, cells were treated with 6.25mM 5a in the presence and absence of 10M
cycloheximide (CHX). Samples were collected at times indicated and levels of p53 were
determined using immunoblotting as described above.
MDR assays:
24

ATPase activity of Pgp was measured using Pgp-Glo assay (Promega) from untreated,
100M Na3VO4-, 200M Verapamil- and 10M 5a compound-treated Pgp reactions. Basal,
verapamil-treated and 5a-treated activity of the Pgp ATPase was calculated according to the
manufacturer’s instructions. ΔRLU= Difference in relative light units.
Reference
(1)
Mansoori, B.; Mohammadi, A.; Davudian, S.; Shirjang, S.; Baradaran, B. The
Different Mechanisms of Cancer Drug Resistance: A Brief Review. Advanced
pharmaceutical bulletin 2017, 7, 339-348.
(2)
quality of life issues. The Malaysian journal of medical sciences : MJMS 2005, 12, 3-5.
(3) Hannun, Y. A. Apoptosis and the dilemma of cancer chemotherapy. Blood
1997, 89, 1845-1853.
Khan, F. A.; Akhtar, S. S.; Sheikh, M. K. Cancer treatment - objectives and
(4)
Gajdács, M.; Spengler, G.; Sanmartín, C.; Marć, M. A.; Handzlik, J.;
Domínguez-Álvarez, E. Selenoesters and selenoanhydrides as novel multidrug resistance
reversing agents: A confirmation study in a colon cancer MDR cell line. Bioorganic &
medicinal chemistry letters 2017, 27, 797-802.
(5)
Structure in Medicinal Chemistry. Chem Rev 2017, 117, 7762-7810.
(6) Sung, J.; Lee, J. Anti-inflammatory activity of butein and luteolin through
Zhuang, C.; Zhang, W.; Sheng, C.; Xing, C.; Miao, Z. Chalcone: A Privileged
suppression of NF κ B activation and induction of heme oxygenase-1. Journal of medicinal
food 2015, 18, 557-564.
(7)
Labib, M. B.; Sharkawi, S. M. Z.; El-Daly, M. Design, synthesis of novel
isoindoline hybrids as COX-2 inhibitors: Anti-inflammatory, analgesic activities and docking
study. Bioorganic Chemistry 2018, 80, 70-80.
(8)
Gaur, R.; Yadav, K. S.; Verma, R. K.; Yadav, N. P.; Bhakuni, R. S. In vivo
anti-diabetic activity of derivatives of isoliquiritigenin and liquiritigenin. Phytomedicine
2014, 21, 415-422.
(9)
Zhang, X.; Rakesh, K. P.; Bukhari, S. N. A.; Balakrishna, M.; Manukumar, H.
M.; Qin, H.-L. Multi-targetable chalcone analogs to treat deadly Alzheimer’s disease: Current
view and upcoming advice. Bioorganic Chemistry 2018, 80, 86-93.
(10) Gan, X.; Hu, D.; Chen, Z.; Wang, Y.; Song, B. Synthesis and antiviral
evaluation of novel 1,3,4-oxadiazole/thiadiazole-chalcone conjugates. Bioorg Med Chem Lett
2017, 27, 4298-4301.
(11) Mirzaei, H.; Emami, S. Recent advances of cytotoxic chalconoids targeting
tubulin polymerization: Synthesis and biological activity. European journal of medicinal
chemistry 2016, 121, 610-639.
(12) Wang, G.; Qiu, J.; Xiao, X.; Cao, A.; Zhou, F. Synthesis, biological evaluation
and molecular docking studies of a new series of chalcones containing naphthalene moiety as
anticancer agents. Bioorganic Chemistry 2018, 76, 249-257.
(13) Sultana, F.; Reddy Bonam, S.; Reddy, V. G.; Nayak, V. L.; Akunuri, R.; Rani
Routhu, S.; Alarifi, A.; Halmuthur, M. S. K.; Kamal, A. Synthesis of benzo[d]imidazo[2,1-
b]thiazole-chalcone conjugates as microtubule targeting and apoptosis inducing agents.
Bioorganic Chemistry 2018, 76, 1-12.
(14) Wang, G.; Peng, Z.; Zhang, J.; Qiu, J.; Xie, Z.; Gong, Z. Synthesis, biological
evaluation and molecular docking studies of aminochalcone derivatives as potential
anticancer agents by targeting tubulin colchicine binding site. Bioorganic Chemistry 2018,
78, 332-340.
25

(15) Fouad, M. M.; El-Bendary, E. R.; suddek, G. M.; Shehata, I. A.; El-Kerdawy,
M. M. Synthesis and in vitro antitumor evaluation of some new thiophenes and thieno[2,3-
d]pyrimidine derivatives. Bioorganic Chemistry 2018, 81, 587-598.
(16) Zhang, E. H.; Wang, R. F.; Guo, S. Z.; Liu, B. An update on antitumor activity
of naturally occurring chalcones. Evidence-based complementary and alternative medicine :
eCAM 2013, 2013, 815621.
(17) Bois, F.; Boumendjel, A.; Mariotte, A. M.; Conseil, G.; Di Petro, A. Synthesis
and biological activity of 4-alkoxy chalcones: potential hydrophobic modulators of P-
glycoprotein-mediated multidrug resistance. Bioorg Med Chem 1999, 7, 2691-2695.
(18) Song, D.-m.; Jung, K.-h.; Moon, J.-h.; Shin, D.-m. Photochemistry of
chalcone and the application of chalcone-derivatives in photo-alignment layer of liquid
crystal display. Opt. Mater. 2003, 21, 667-671.
(19) Cong, H.; Zhao, X. H.; Castle, B. T.; Pomeroy, E. J.; Zhou, B.; Lee, J.; Wang,
Y.; Bian, T. F.; Miao, Z. Y.; Zhang, W. N.; Sham, Y. Y.; Odde, D. J.; Eckfeldt, C. E.; Xing,
C. G.; Zhuang, C. L. An Indole-Chalcone Inhibits Multidrug-Resistant Cancer Cell Growth
by Targeting Microtubules. Mol Pharmaceut 2018, 15, 3892-3900.
(20) Khan, N. S.; Khan, P.; Ansari, M. F.; Srivastava, S.; Hasan, G. M.; Husain,
M.; Hassan, M. I. Thienopyrimidine-Chalcone Hybrid Molecules Inhibit Fas-Activated
Serine/Threonine Kinase: An Approach To Ameliorate Antiproliferation in Human Breast
Cancer Cells. Mol. Pharm. 2018, 15, 4173-4189.
(21) Takac, P.; Kello, M.; Pilatova, M. B.; Kudlickova, Z.; Vilkova, M.;
Slepcikova, P.; Petik, P.; Mojzis, J. New chalcone derivative exhibits antiproliferative
potential by inducing G2/M cell cycle arrest, mitochondrial-mediated apoptosis and
modulation of MAPK signalling pathway. Chem Biol Interact 2018, 292, 37-49.
(22) Sultana, F.; Bonam, S. R.; Reddy, V. G.; Nayak, V. L.; Akunuri, R.; Routhu,
S. R.; Alarifi, A.; Halmuthur, M. S. K.; Kamal, A. Synthesis of benzo[d]imidazo[2,1-
b]thiazole-chalcone conjugates as microtubule targeting and apoptosis inducing agents.
Bioorg Chem 2018, 76, 1-12.
(23) Zhu, M.; Wang, J.; Xie, J.; Chen, L.; Wei, X.; Jiang, X.; Bao, M.; Qiu, Y.;
Chen, Q.; Li, W. Design, synthesis, and evaluation of chalcone analogues incorporate α, β-
Unsaturated ketone functionality as anti-lung cancer agents via evoking ROS to induce
pyroptosis. Eur J Med Chem 2018, 157, 1395-1405.
(24) Park, S.; Kim, E. H.; Kim, J.; Kim, S. H.; Kim, I. Biological evaluation of
indolizine-chalcone hybrids as new anticancer agents. Eur J Med Chem 2018, 144, 435-443.
(25) Li, P. H.; Jiang, H.; Zhang, W. J.; Li, Y. L.; Zhao, M. C.; Zhou, W.; Zhang, L.
Y.; Tang, Y. D.; Dong, C. Z.; Huang, Z. S.; Chen, H. X.; Du, Z. Y. Synthesis of carbazole
derivatives containing chalcone analogs as non-intercalative topoisomerase II catalytic
inhibitors and apoptosis inducers. Eur J Med Chem 2018, 145, 498-510.
(26) Gil, H. N.; Koh, D.; Lim, Y.; Lee, Y. H.; Shin, S. Y. The synthetic chalcone
derivative 2-hydroxy-3 ',5,5 '-trimethoxychalcone induces unfolded protein response-
mediated apoptosis in A549 lung cancer cells. Bioorg Med Chem Lett 2018, 28, 2969-2975.
(27) Silva, G.; Marins, M.; Chaichanasak, N.; Yoon, Y.; Fachin, A. L.; Pinhanelli,
V. C.; Regasini, L. O.; Dos Santos, M. B.; Ayusso, G. M.; Marques, B. C.; Wu, W. W.; Phue,
J. N.; Shen, R. F.; Baek, S. J. Trans-chalcone increases p53 activity via DNAJB1/HSP40
induction and CRM1 inhibition. PloS one 2018, 13, e0202263.
(28) Seba, V.; Silva, G.; Santos, M.; Baek, S.; França, S.; Fachin, A.; Regasini, L.;
Marins, M. Chalcone Derivatives 4′-Amino-1-Naphthyl-Chalcone (D14) and 4′-Amino-4-
Methyl-1-Naphthyl-Chalcone (D15) Suppress Migration and Invasion of Osteosarcoma Cells
Mediated by p53 Regulating EMT-Related Genes. INT J MOL SCI 2018, 19, 2838.
26

(29) Ducki, S.; Forrest, R.; Hadfield, J. A.; Kendall, A.; Lawrence, N. J.; McGown,
A. T.; Rennison, D. Potent antimitotic and cell growth inhibitory properties of substituted
chalcones. Bioorg. Med. Chem. Lett. 1998, 8, 1051-1056.
(30) Lawrence, N. J.; Patterson, R. P.; Ooi, L.-L.; Cook, D.; Ducki, S. Effects of α-
substitutions on structure and biological activity of anticancer chalcones. Bioorganic &
medicinal chemistry letters 2006, 16, 5844-5848.
(31) Smith, C. D.; Rosocha, G.; Mui, L.; Batey, R. A. Investigation of Substituent
Effects on the Selectivity of 4π-Electrocyclization of 1, 3-Diarylallylic Cations for the
Formation of Highly Substituted Indenes. The Journal of organic chemistry 2010, 75, 4716-
4727.
(32) Yin, B.-T.; Yan, C.-Y.; Peng, X.-M.; Zhang, S.-L.; Rasheed, S.; Geng, R.-X.;
Zhou, C.-H. Synthesis and biological evaluation of α-triazolyl chalcones as a new type of
potential antimicrobial agents and their interaction with calf thymus DNA and human serum
albumin. European journal of medicinal chemistry 2014, 71, 148-159.
(33) Mehnaz, S.; Saleem, R. S. Z.; Yameen, B.; Pianet, I.; Schnakenburg, G.;
Pietraszkiewicz, H.; Valeriote, F.; Josten, M.; Sahl, H.-G.; Franzblau, S. G.; Gross, H.
Lahorenoic Acids A–C, ortho-Dialkyl-Substituted Aromatic Acids from the Biocontrol Strain
Pseudomonas aurantiaca PB-St2. Journal of Natural Products 2013, 76, 135-141.
(34) Hanif, A.; Sardar, A.; Alazmi, M.; Tariq, H.; Emwas, A. H.; Gao, X.; Chotana,
G. A.; Zaib Saleem, R. S. A Novel One‐ Pot Three‐ Component Reaction for Rapid Access
of Arylidene 2‐ Aminoimidazolone Derivatives. ChemistrySelect 2019, 4, 1753-1756.
(35) Shahid, I.; Rizwan, M.; Baig, D. N.; Saleem, R. S.; Malik, K. A.; Mehnaz, S.
Secondary metabolites production and plant growth promotion by Pseudomonas chlororaphis
and P. aurantiaca strains isolated from cactus, cotton, and para grass. MicrobiolBiotechnol
2017, 27, 480-491.
(36) Batool, S.; Ullah, S.; Tabassum, S.; Bilal, A.; Faisal, A.; Saleem, R. S. Z.;
Ahmad, M. S. Effect of Extraction Methodologies on Antioxidant Potential and Synergistic
Anticancer Behavior of Leaf Extract of Polygonum amplexicaule Against HCT-116 Cells.
Iranian Journal of Science and Technology, Transactions A: Science, 1-6.
(37) Saleem, R. S. Z.; Tepe, J. J. A concise total synthesis of hymenialdisine.
Tetrahedron Letters 2015, 56, 3011-3013.
(38) Iftikhar, S.; Khan, S.; Bilal, A.; Manzoor, S.; Abdullah, M.; Emwas, A.-H.;
Sioud, S.; Gao, X.; Chotana, G. A.; Faisal, A. Synthesis and evaluation of modified chalcone
based p53 stabilizing agents. Bioorganic & medicinal chemistry letters 2017, 27, 4101-4106.
(39) Manzoor, S.; Bilal, A.; Khan, S.; Ullah, R.; Iftikhar, S.; Emwas, A.-H.;
Alazmi, M.; Gao, X.; Jawaid, A.; Saleem, R. S. Z. Identification and characterization of
SSE15206, a microtubule depolymerizing agent that overcomes multidrug resistance.
Scientific reports 2018, 8, 3305.
(40) Farrukh, S. U. B.; Javed, I.; Ather, A. Q.; Emwas, A.-H.; Alazmi, M.; Gao, X.;
Chotana, G. A.; Davis, T. P.; Ke, P. C.; Saleem, R. S. Z. Synthesis and identification of novel
pyridazinylpyrazolone based diazo compounds as inhibitors of human islet amyloid
polypeptide aggregation. Bioorganic Chemistry 2019, 84, 339-346.
(41) Jawad Nasim, M.; Ali, W.; Domínguez-Álvarez, E.; da Silva Júnior, E. N.;
Saleem, R. S. Z.; Jacob, C.: Chapter 10 Reactive Selenium Species: Redox Modulation,
Antioxidant, Antimicrobial and Anticancer Activities. In Organoselenium Compounds in
Biology and Medicine: Synthesis, Biological and Therapeutic Treatments; The Royal Society
of Chemistry, 2018; pp 277-302.
(42) Saleem, R. S. Z.; Lansdell, T. A.; Tepe, J. J. Synthesis and evaluation of
debromohymenialdisine-derived Chk2 inhibitors. Bioorganic & Medicinal Chemistry 2012,
20, 1475-1481.
27

(43) Nguyen, T. N. T.; Saleem, R. S. Z.; Luderer, M. J.; Hovde, S.; Henry, R. W.;
Tepe, J. J. Radioprotection by Hymenialdisine-Derived Checkpoint Kinase 2 Inhibitors. ACS
Chemical Biology 2012, 7, 172-184.
(44) Rendekova, J.; Vlasakova, D.; Arsenyan, P.; Vasiljeva, J.; Nasim, M. J.;
Witek, K.; Dominguez-Alvarez, E.; Zeslawska, E.; Manikova, D.; Tejchman, W. The
Selenium-Nitrogen Bond as Basis for Reactive Selenium Species with Pronounced
Antimicrobial Activity. Current Organic Synthesis 2017, 14, 1082-1090.
(45) Park, K. K.; Jeong, J. Facile synthesis of regio-isomeric naphthofurans and
benzodifurans. Tetrahedron 2005, 61, 545-553.
(46) Milcent, R.; Malanda, J. C.; Barbier, G.; Vaissermann, J. Synthesis of ethyl
2‐ aminodihydro‐ 5‐ pyrimidinecar‐ boxylate derivatives and 3, 7‐ diethoxycarbonyl‐ 4,
6‐ dihydro‐ 2, 4, 6, 8‐ tetraaryl‐ 1H‐ pyrimido [1, 2‐ a] pyrimidines. Journal of
heterocyclic chemistry 1997, 34, 329-336.
28

Graphical abstract
29