Synthesis of some coumarinyl chalcones and their antiproliferative activity against breast cancer cell lines

Article abstract of DOI:10.2174/157018011794839475

A series of coumarinyl chalcones derivatives were synthesized and evaluated for their antiproliferative activities on three different breast cancer cell lines (MDA-MB231, MDA-MB468, MCF7) and one non-cancer breast epithelial cell line (184B5). The coumarinyl derivatives exhibited anticancer activity against breast cancer cell lines at a micromolar range. A structure-activity relationship (SAR) analysis was performed by studying the effect of substituents on their antiproliferative activities. One of the compound 3i bearing methoxy substitutions at the R₁; R₂ and R₃ positions of the phenyl ring showed comparable potency to the reference drug cisplatin as well as a two-fold higher selectivity for the breast cancer cell lines than 184B5 cells.

Full text of DOI:10.2174/157018011794839475

308
Letters in Drug Design & Discovery, 2011, 8, 308-311
Synthesis of Some Coumarinyl Chalcones and their Antiproliferative
Activity Against Breast Cancer Cell Lines
Kuldeep Patel1, Chandrabose Karthikeyan1, Viswas Raja Solomon2,3, N.S. Hari Narayana
Moorthy1,4, Hoyun Lee2,3, Kapendra Sahu1, Girdhar Singh Deora1 and Piyush Trivedi*,1
1School of Pharmaceutical Sciences, Rajiv Gandhi Technical University, Airport Bypass Road, Gandhi Nagar,
Bhopal-462036 (M.P.), India
2Tumour Biology Group, Northeastern Ontario Regional Cancer Program at the Sudbury, Canada and Regional
Hospital, 41 Ramsey Lake Road, Sudbury, Ontario, P3E 5J1, Canada
3Department of Biology, Laurentian University, Sudbury, Ontario P3E 2C6, Canada
4Department of Chemistry & Biochemistry, Faculty of Sciences, University of Porto, 687, Rua de Campo Alegre,
Porto-4169-007, Portugal
Received August 20, 2010: Revised January 02, 2011: Accepted January 10, 2011
Abstract: A series of coumarinyl chalcones derivatives were synthesized and evaluated for their antiproliferative activi-
ties on three different breast cancer cell lines (MDA-MB231, MDA-MB468, MCF7) and one non-cancer breast epithelial
cell line (184B5). The coumarinyl derivatives exhibited anticancer activity against breast cancer cell lines at a micromolar
range. A structure-activity relationship (SAR) analysis was performed by studying the effect of substituents on their anti-
proliferative activities. One of the compound 3i bearing methoxy substitutions at the R1, R2 and R3 positions of the phenyl
ring showed comparable potency to the reference drug cisplatin as well as a two-fold higher selectivity for the breast can-
cer cell lines than 184B5 cells.
Keywords: Coumarinyl chalcones, Breast cancer cell lines, Antiproliferative activity, 184B5.
INTRODUCTION
activities, including antileishmanial, antiinflammatory, an-
timitotic, modulation of P-glycoprotein-mediated multidrug
resistance, and antimalarial activities, etc [17-22]. Naturally
occurring chalcones are mostly in the hydroxylated forms;
such as, butein, licochalcone-A, isoliquiritigenin, xanthoan-
gelol and flavokawain A. They have been shown to arrest
cell cycle progression, induce apoptosis, and inhibit tumor
promotion and metastasis in various cancer cells [18, 23-27].
There are also several reports on the antiproliferative activity
of chalcones against breast cancer cell lines [4, 28-30].
Cancer is the second leading cause of human death after
cardiovascular diseases worldwide and more than 70% of all
cancer deaths occurred in developing and under developed
countries [1,2]. There is a continuous rise of deaths from
various cancers worldwide and with an estimated 12 million
deaths in 2030 [3]. Among the cancers, breast cancer is the
most common cancer among women after skin cancer, and it
is also the second leading cause of cancer death (after lung
cancer) in women [4,5]. Many different therapeutic strategies
are currently available for cancer treatments, including che-
motherapy and radiotherapy. However, systemic toxicity of
the chemotherapeutic agents and emergence of drug resistant
tumors limit the successful outcomes in most cases. Thus,
there is a continuous need for the development of novel anti-
cancer agents for the treatment of (breast) cancer.
Encouraged by these findings and our continuing inter-
ests in the exploration of novel heterocyclic scaffolds for
anticancer activity [31,32], we have synthesized a number of
coumarinyl chalcones and investigated their antiproliferative
activities against three breast cancer cell lines.
The synthesis of the coumarinyl chalcones (3a-3j) was
accomplished by a two-step procedure outlined in the
Scheme 1, as described previously [33]. The first step in-
volved synthesis of the precursor 4-hydroxy-3-acetyl cou-
marin (2) by reacting 4-hydroxy coumarin (1) with phospho-
rous oxychloride and glacial acetic acid. The second step
involved the Knoevenagel condensation between the same
molar amount of 4-hydroxy-3-acetyl coumarin (3), and sub-
stituted benzaldehydes with chloroform in the presence of
piperidine catalyst. The spectral data of the synthesized
compounds (IR, 1H NMR and Mass) were in good agreement
with their structure [34].
Recently, many heterocyclic and non-heterocyclic scaf-
folds have been investigated for antiproliferative activity
against various tumor cell lines [6-14]. Among them chal-
cones is one of the most promising classes of compounds
that exhibit anticancer activities along with other pharmacol-
ogical activities. Chalcones are structurally simple com-
pounds of the flavonoid family and are present in a variety of
plant species [15,16]. Chemically, these are 1,3-diphenyl-2-
propen-1-one and have reported a wide range of biological
*Address correspondence to this author at the School of Pharmaceutical
Sciences, Rajiv Gandhi Technical University, Airport Bypass Road, Gandhi
Nagar, Bhopal-462036 (M.P.), India; Tel: +91-755-2678883; Fax: +91-755-
2742006; E-mail: prof_piyushtrivedi@yahoo.com, piyush.trivedi@rgtu.net
All the compounds synthesized were evaluated for their
cytotoxic effects on three breast cancer cell lines, MDA-
MB468, MDA-MB231 and MCF7, and one non-cancer
1570-1808/11 $58.00+.00
© 2011 Bentham Science Publishers Ltd.

Synthesis of Some Coumarinyl Chalcones
Letters in Drug Design & Discovery, 2011, Vol. 8, No. 4 309
OH
O
O
OH
OH
O
R4
R3
a
b
O
O
O
O
O
R1
(1)
(2)
R2
4-hydroxy-2H-chromen-2-one
(3a-j)
3-acetyl-4-hydroxy-2H-chromen-2-one
Scheme 1. Synthesis of the 3-cinnamoyl-4-hydroxy-2H-chromen-2-ones.
Reagents and Conditions: “a” denotes Glacial acetic acid, POCl3, reflux; “b” is substituted benzaldehydes, CHCl3, piperidine (Catalytic),
reflux.
Table 1. Structure and Cytotoxic Activities of the Synthesized Compounds (3a-j)
OH
O
R4
R3
O
O
R1
R2
GI50 a,b (ꢀM)
Compounds
R1
R2
R3
R4
MDA-MB231
MDA-MB468
MCF7
184B5
3a
3b
3c
3d
3e
3f
H
Cl
H
H
H
H
H
H
H
67.58±0.94
102.06±2.01
68.95±0.87
61.33±0.92
58.75±0.78
111.51±1.97
62.60±0.85
102.33±1.98
22.11±0.21
53.64±0.65
23.65±0.23
113.89±1.36
65.34±0.88
44.94±0.65
47.89±0.72
110.82±1.33
117.54±1.69
58.48±0.48
169.25±1.81
41.08±0.52
84.41±1.24
31.02±0.45
80.59±1.25
90.72±1.75
84.60±1.32
67.83±1.01
63.45±0.99
90.72±1.87
75.43±1.36
98.27±1.48
25.86±0.25
74.66±1.23
25.77±0.38
65.56±1.32
60.09±1.01
39.19±0.85
75.09±0.98
152.08±2.01
123.26±1.36
75.88±0.98
120.22±1.23
47.41±0.69
80.36±1.58
25.54±0.35
H
Cl
H
Cl
H
H
Cl
H
NO2
H
H
H
H
NO2
N(CH3)2
OCH3
OCH3
OCH3
H
3g
3h
3i
H
H
H
H
OCH3
OCH3
OCH3
H
OCH3
H
H
3j
OCH3
Cisplatin
a GI50 values were calculated from Sigmoidal dose response curves (variable slope), which were generated with GraphPad Prism V. 4.02 (GraphPad Software Inc.). b Values are the
mean of triplicates of at least two independent experiments.
breast epithelial cell lines, 184B5. Each compound stored at
20 mM was diluted from 200 ꢀM to 0.0128 ꢀM by five-fold
serial dilutions. Cells were treated with each compound for
48 h, followed by measuring cell growth/proliferation by
SRB-based spectrophotometry as described previously
[35,36]. The reading of SRB staining is known to accurately
reflect the levels of total cellular macromolecules/cell
growth/proliferation [36]. The GI50 concentration for each
compound was calculated with reference to a control sample,
which represents the concentration that results in a 50% de-
crease in cell growth/proliferation after 48 h incubation in
the presence of drugs. For each compound, 50% growth in-
hibition (GI50) was calculated from Sigmoidal dose-response
curves and presented in Table 1. The well known anticancer
drug cisplatin was used as a reference compound.
structure of the compounds and the anticancer activity
against breast cancer cell lines showed that the compound 3i
in this series has the highest anticancer activity against all
three breast cancer cell lines examined. Structurally, this
compound possesses methoxy substitutions at the R1, R2 and
R3 positions on the phenyl ring. Its antiproliferative activity
against MDA-MB231 (22.11±0.21 ꢀM) and MCF7
(25.86±0.25 ꢀM) cell lines was comparable to the reference
compound cisplatin, of which GI50 were 23.65±0.23 ꢀM and
25.77±0.38 ꢀM against MDA-MB231 and MCF7, respec-
tively. Furthermore, the compound also showed a two-fold
higher cytotoxic effect on the three cancer cell lines than the
non-cancer 184B5 breast epithelial cell line. However, simi-
lar substitutions at the R2, R3 and R4 positions of the phenyl
ring (compound 3j) showed only moderate potency against
the breast cancer cell lines examined. Methoxy substitutions
at the R2 and R3 positions of the phenyl ring (compound 3h)
resulted in the loss of activity, suggesting the importance of
the 2 methoxy group for the anticancer activity. The com-
Anticancer activity screening of coumarinyl chalcones
against the three breast cancer cell lines revealed that all of
the compounds exhibited growth inhibitory activity (GI50) at
micromolar concentrations. The relationship between the

310 Letters in Drug Design & Discovery, 2011, Vol. 8, No. 4
Patel et al.
pound 3j showed a two-fold less activity than 3i, and a two-
fold greater anticancer activity than 3h in terms of MDA-
MB231and MDA-MB468 cell growth inhibition.
har Singh Deora, and Kapendra Sahu wish to thank AICTE,
New Delhi for a postgraduate fellowship. C. Karthikeyan
wishes to thank CSIR, New Delhi for providing a Senior
Research Fellowship. V. R. S. is a recipient of a postdoctoral
fellowship from the Ontario Ministry of Research and Inno-
vation.
The compounds with a chlorine substitution on the
phenyl ring (R1 or/and R3 positions) (3c and 3d) of the cou-
marinyl chalcone possess a significant anticancer activity
against MDA-MB231 and only a moderate activity against
MCF7. These compounds showed 1-3 folds higher MDA-
MB468 inhibitory activity than the unsubstituted compound
(3a), and 1-3 times less activity than cisplatin. However,
compound 3d was more selective for breast cancer cell lines
than non-cancer breast epithelial cell line 184B5. In contrast
compound 3c showed greater cytotoxicity on the 184B5 non-
cancer cell line (39.19±0.85 ꢀM), compared to cancer cell
lines (68.95, 44.94, and 84.60 ꢀM against MDA-MB231,
MDA-MB468, and MCF7, respectively). Substitution of
chlorine atom at the R1 position on the phenyl ring (3b) re-
sulted in decreased antiproliferative activity against MDA-
MB231 and MCF7 cell lines. The unsubstituted phenyl de-
rivative (3a) showed a better activity than the para nitro (3e)
substituted phenyl derivative in all breast cancer cell lines.
Nitro substitution at the R2 position (3e) rendered a moder-
ate anticancer activity against MDA-MB231 and MCF7, and
a decrease in cytotoxicity against MDA-MB468 and 184B5
cell lines. The compound with dimethyl amino moiety at R3
position on the phenyl ring (3g) showed only a moderate
activity against the entire cell lines examined.
SUPPLEMENTARY MATERIAL
Supplementary material is available on the publishers
Web site along with the published article.
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