Letters in Drug Design & Discovery, 2011, 8, ???-???
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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 1
School of Pharmaceutical Sciences, Rajiv Gandhi Technical University, Airport Bypass Road, Gandhi Nagar, Bhopal-462036 (M.P.), India 2
Tumour Biology Group, Northeastern Ontario Regional Cancer Program at the Sudbury
Regional Hospital, 41 Ramsey Lake Road, Sudbury, Ontario, P3E 5J1, Canada 3
Department of Biology, Laurentian University, Sudbury, Ontario P3E 2C6, Canada
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Department 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 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. Keywords: ????????????????????????????????????.
INTRODUCTION 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 chemotherapy 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 anticancer agents for the treatment of (breast) cancer. Recently, many heterocyclic and non-heterocyclic scaffolds have been investigated for antiproliferative activity against various tumor cell lines [6-14]. Among them chalcones is one of the most promising classes of compounds that exhibit anticancer activities along with other pharmacological activities. Chalcones are structurally simple compounds of the flavonoid family and are present in a variety of plant species [15,16]. Chemically, these are 1,3-diphenyl-2propen-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: +91-7552742006; E-mail:
[email protected],
[email protected] 1570-1808/11 $58.00+.00
activities, including antileishmanial, antiinflammatory, antimitotic, 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, xanthoangelol 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]. Encouraged by these findings and our continuing interests 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 involved synthesis of the precursor 4-hydroxy-3-acetyl coumarin (2) by reacting 4-hydroxy coumarin (1) with phosphorous 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 substituted 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]. All the compounds synthesized were evaluated for their cytotoxic effects on three breast cancer cell lines, MDA© 2011 Bentham Science Publishers Ltd.
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Letters in Drug Design & Discovery, 2011, Vol. 8, No. 4
Patel et al.
OH
OH
O
a O
OH
O R4
b O
O
(1)
O
O
O
R1
(2)
R3 R2
4-hydroxy-2H-chromen-2-one 3-acetyl-4-hydroxy-2H-chromen-2-one
(3a-j)
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
O
O
R1
R3 R2 GI50 a,b (M)
Compounds
R1
R2
R3
R4 MDA-MB231
MDA-MB468
MCF7
184B5
3a
H
H
H
H
67.58±0.94
113.89±1.36
80.59±1.25
65.56±1.32
3b
Cl
H
H
H
102.06±2.01
65.34±0.88
90.72±1.75
60.09±1.01
3c
H
H
Cl
H
68.95±0.87
44.94±0.65
84.60±1.32
39.19±0.85
3d
Cl
H
Cl
H
61.33±0.92
47.89±0.72
67.83±1.01
75.09±0.98
3e
H
NO2
H
H
58.75±0.78
110.82±1.33
63.45±0.99
152.08±2.01
3f
H
H
NO2
H
111.51±1.97
117.54±1.69
90.72±1.87
123.26±1.36
3g
H
H
N(CH3)2
H
62.60±0.85
58.48±0.48
75.43±1.36
75.88±0.98
3h
H
OCH3
OCH3
H
102.33±1.98
169.25±1.81
98.27±1.48
120.22±1.23
3i
OCH3
OCH3
OCH3
H
22.11±0.21
41.08±0.52
25.86±0.25
47.41±0.69
3j
H
OCH3
OCH3
OCH3
53.64±0.65
84.41±1.24
74.66±1.23
80.36±1.58
23.65±0.23
31.02±0.45
25.77±0.38
25.54±0.35
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.
MB468, MDA-MB231 and MCF7, and one non-cancer 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% decrease in cell growth/proliferation after 48 h incubation in the presence of drugs. For each compound, 50% growth inhibition (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. 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 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, respectively. 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, similar 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
Synthesis of Some Coumarinyl Chalcones
the 2 methoxy group for the anticancer activity. The compound 3j showed a two-fold less activity than 3i, and a twofold greater anticancer activity than 3h in terms of MDAMB231and MDA-MB468 cell growth inhibition. The compounds with a chlorine substitution on the phenyl ring (R1 or/and R3 positions) (3c and 3d) of the coumarinyl chalcone possess a significant anticancer activity against MDA-MB231 and only a moderate activity against MCF7. These compounds showed 1-3 folds higher MDAMB468 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 noncancer cell line (39.19±0.85 M), compared to cancer cell lines (68.95, 44.94, and 84.60 M against MDAMB231, MDA-MB468, and MCF7, respectively). Substitution of chlorine atom at the R1 position on the phenyl ring (3b) resulted in decreased antiproliferative activity against MDA-MB231 and MCF7 cell lines. The unsubstituted phenyl derivative (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 of the (3e) rendered a moderate anticancer activity against MDAMB231 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. From SAR analysis, it is evident that the antiproliferative activity of the coumarinyl chalcones against MDA-MB231 cell lines was predominantly influenced by the structural variation in the phenyl ring, whereas the activity of title compounds against MDA-MB468 and MCF7 did not appear to be influenced by changes in their molecular structure except for compound 3i. Compound 3i bearing methoxy groups at the 2,3,4 positions showed exceptional antiproliferative potency and selectivity in comparison to other compounds in this series. Hence, it would be an ideal starting point for further exploration of this series of compounds as anticancer agents. In summary, the present investigation has revealed coumarinyl chalcones have the potential as lead compounds for the development of effective antiproliferative agents for the treatment of (breast) cancer. Compound 3i with a methoxy substitution at the 2,3,4 positions of the phenyl ring showed an antiproliferative activity comparable to the reference drug cisplatin, and also exhibited better selectivity for breast cancer cells, compared to the 184B5 non-cancer breast cell line. The SAR studies revealed that the antiproliferative activity of title compounds against MDA-MB231 cell lines could be improved by manipulating the structural pattern on the phenyl ring. Taken together, our data suggest that coumarinyl chalcones can be promising lead compounds in he development of effective cancer therapeutics.
Letters in Drug Design & Discovery, 2011, Vol. 8, No. 4
New Delhi, India for the NMR spectral analysis of the compounds used in this study. The authors Kuldeep Patel, Girdhar 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 Innovation. SUPPLEMENTARY MATERIAL Supplementary material is available on the publishers Web site along with the published article. REFERENCES [1]
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