Cytotoxic potential of dispirooxindolo/acenaphthoquino andrographolide derivatives against MCF-7 cell line

Article abstract of DOI:10.1039/c4md00469h

Dispiro andrographolide derivatives have been prepared from isatin/acenaphthoquinone, N-benzyl glycine and andrographolide via azomethine ylide cycloaddition reaction. The cytotoxic effect of the synthesized molecules has been studied against MCF-7 breast

Full text of DOI:10.1039/c4md00469h

View Article Online
View Journal
MedChemComm
Accepted Manuscript
This article can be cited before page numbers have been issued, to do this please use: D. Chakraborty,
A. Maitya, C. K. Jain, A. Hazra, Y. P. Bharitkar, T. Jha, H. Majumder, S. R. Choudhury and N. B. Mondal,
This is an Accepted Manuscript, which has been through the
Royal Society of Chemistry peer review process and has been
accepted for publication.
Accepted Manuscripts are published online shortly after
acceptance, before technical editing, formatting and proof reading.
Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available.
You can find more information about Accepted Manuscripts in the
Information for Authors.
Please note that technical editing may introduce minor changes
to the text and/or graphics, which may alter content. The journal’s
standard Terms & Conditions and the Ethical guidelines still
apply. In no event shall the Royal Society of Chemistry be held
responsible for any errors or omissions in this Accepted Manuscript
or any consequences arising from the use of any information it
contains.
www.rsc.org/medchemcomm

Medicinal Chemistry Communications
Page 1 of 6Journal Name
Dynamic Article Links ►
View Article Online
DOI: 10.1039/C4MD00469H
Cite this: DOI: 10.1039/c0xx00000x
www.rsc.org/xxxxxx
ARTICLE TYPE
Cytotoxic potential of di-spirooxindolo/acenaphthoquino
andrographolide derivatives against MCF7 cell lines
Debanjana Chakrabortya, Arindam Maitya, Chetan K. Jainb,c, Abhijit Hazraa, Yogesh P. Bharitkara,
Tarun Jhad, Hemanta K. Majumderb, Susanta Roychoudhuryc, Nirup B. Mondala*
5
Received (in XXX, XXX) Xth XXXXXXXXX 20XX, Accepted Xth XXXXXXXXX 20XX
DOI: 10.1039/b000000x
Di-spiro andrographolide derivatives have been prepared from isatin/ acenaphthoquinone, N-benzyl
glycine and andrographolide via azomethine ylide cycloaddition reaction. Cytotoxic effect of the
synthesized molecules has been studied against MCF-7 breast cancer cell lines. The compounds induced
10 apoptotic cell death as revealed by increased labeling with Annexin-V, decreased polarization of cell
mitochondria, and increased reactive oxygen species production. Using FACS and western blot analysis,
the compounds were observed to block the cell cycle at S and G2-M phases by causing DNA damage that
halted replication and ultimately caused apoptosis. Activation of caspases 7 and 9 suggested that caspase
pathways were involved in inducing apoptosis.
drawback of andrographolide is poor water solubility making it
15 Introduction
difficult to prepare formulations for clinical use. So, various semi
synthetic analogues are being developed and evaluated in order to
find out a better lead.14-18Jada et al. recently reported that 14-
50 acetylandrographolide is more potent in vitro against leukemia
compared to andrographolide.15, 16 Another report identified DRF
3188, a novel derivative of andrographolide having α,β-
unsaturated ester side chain at C14, to have better anti-cancer
activity than the parent molecule.17 Encouraged by these reports,
55 we prepared a few dispiro analogues of andrographolide via
azomethine ylide cycloaddition.19The results of the biological
evaluation of these compounds20 encouraged us further to build a
library of dispiro analogues containing both spiro-oxindole and
pyrrolidine/pyrrolizidine rings attached to andrographolide.21
60 Preliminary bio-evaluation indicated the promise held out by N-
benzyl glycine derivatives that needed to be further explored. In
the present study, we evaluated the anticancer efficacy of some of
these analogues of andrographolide which showed good MTT
value for MCF-7 breast cancer cell line. As apoptosis is the
65 physiologically desired pathway of cell death by the anticancer
agents22, 23 we wanted to explore the involvement of apoptosis in
cell death induced by the andrographolide derivatives.
The human race has hugely benefited from drugs discovered from
different natural sources, especially plants. Recent studies
conducted by the World Health Organization (WHO) have
brought out the fact that about 80% of the world’s population rely
20 on traditional medicine.1 At present about 121 drugs prescribed in
USA trace their origin to natural resources, 90 of which either
directly or indirectly come from plant kingdoms.2 Between the
years 1981-2006, about a hundred anticancer agents have been
developed, of which twenty five are natural product derivatives,
25 eighteen are natural product mimics, eleven candidates are
derived from a natural product pharmacophore, and nine are pure
natural products.3
Breast cancer is a very common cancer in women both in the
developed and less developed world and is the second leading
30 cause of cancer death in women. Worldwide over 508,000
women are estimated to have died in 2011 due to breast cancer.4
Although breast cancer is thought to be a disease of the
developed world, almost 50% of breast cancer cases and 58% of
deaths occur in less developed countries.5 It is therefore of
35 immense interest to search for new drug candidates from the plant
kingdom to tackle this menace.
The herb Andrographis paniculata Nees (Acanthaceae) is popular
in India, China and other Asian countries due to its different uses
in traditional medicine. The metabolites isolated from this herb
40 are mainly diterpenoids, flavonoids and sterols. The major
constituent is andrographolide (1), a labdane diterpene. Extracts
of this herb and its various constituents are reported to have a
broad range of biological activities.6-9In the recent past, potent
anti-cancer activities of andrographolide have been described.10-13
45 However, despite its impressive biological activities; the major
Results and discussion
Chemistry
70 As described earlier that the dispiro products were derived from
Andrographolide (1, isolated from A. paniculata in the usual
way), 24-26 isatin derivatives (2 a-k) or acenapthoquinone (3), and
N-benzyl glycine via azomethine ylide cycloaddition19 which
were subsequently crystallized from acetonitrile-benzene
75 (Scheme 1) and characterized by spectral analysis.21
This journal is © The Royal Society of Chemistry [year]
[journal], [year], [vol], 00–00 | 1

Medicinal Chemistry Communications
Page 2 of 6
View Article Online
DOI: 10.1039/C4MD00469H
R3
R2
N
O
The
test
45 phosphatidylserine
compounds
cause
externalization
of
R1
O
O
O
O
R1
N
R2
Annexin V FITC and PI double-labeling studies were used for the
detection of phosphatidylserine externalization. Flow cytometric
analysis showed that MCF7 cells treated with test compounds
showed higher number of annexin V positive cells than control
50 (Figure-1). The percentage of viable cells was significantly low
for all the samples. The total numbers of annexin V binding in
MCF7 cells detectable for 5(b, c, i, g) and 8 exposures are 20.3,
27.2, 23.1, 37.7 and 43.2% respectively after 24 h, and 20.0, 91.9,
61.9, 98.3, and 99.6% respectively after 48 h. The percentage of
55 PI positive cells for these compounds (upper left quadrant) at
baseline were minimal and remained comparable at 24 and 48 h,
being 0.5, 0.9, 0.6, 0.9, and 0.3% in the former case, and 2.2, 0.2,
1.4, 0.0, and 0.0% in the latter, implying negligible occurrence of
necrosis. Taken all together, these compounds cause
60 externalization of phosphatidyl serine, indicative of apoptosis.
N
R3
HO
2 a-k
H
N
COOH
Ph
MW, 15 min
HO
HO
H
5 a-k
O
5a: R1=R2=R3=H
5e: R1=F; R2=R3=H
5i: R1=R2=H; R3=F
5j: R1=R2=H; R3=Br
5k: R1=R3=H; R2=Ph
HO
O
5b: R1=Me; R2=R3=H
5c: R1=Cl; R2=R3=H
5d: R1=I; R2=R3=H
5f: R1=OMe; R2=R3=H
5g: R1=Br; R2=R3=H
5h: R1=R3=Me; R2=H
O
O
O
HO
HO
O
O
H
1
N
HO
3
H
N
COOH
Ph
MW, 15 min
HO
HO
H
8
Scheme 1. Di-spiro andrographolide derivatives (5a-k and 8)
Biological studies
Previously the cytotoxic efficacies of different di-spiropyrrolidino
and dispiropyrrolizidino oxindole andrographolide analogues
prepared by the cycloaddition of azomethine ylides of sarcosine
or proline (described herein as sarcosine and proline series
respectively) at 12,13 double bond of andrographolide were
evaluated by our group.20 The results encouraged us to build a
5
10 library of similar dispiro derivatives by using N-benzyl glycine as
amino acid component and carry out their biological evaluation.
The present communication describes the anticancer role of these
analogues, five of which displayed good MTT values against
MCF-7 breast cancer cell line.
15
Cytotoxicity of andrographolide derivatives (N-benzyl glycine
series) on cancer cell lines
Cell proliferation inhibition was tested by MTT assay following
treatment of cells with 12 derivatives (N-benzyl glycine series)
20 and their cytotoxic potential was tested in MCF 7 (breast
carcinoma), HepG-2 (hepatocellular carcinoma), HeLa (cervical
carcinoma), A431 (epidermoid carcinoma), and CHO (Chinese
hamster ovary) cell lines.21 Out of the 12 derivatives, compounds
5b, 5c, 5g, 5i and 8 were found to be the most potent in different
25 cancer cell lines. These compounds showed very promising
activity particularly in MCF7 cell line, having IC50 values
10.9 0.4, 12.7 0.1, 12.8 0.9, 13.7 1.7 and 8.9 1.6 µM
respectively after 72 h incubation. Based on the results, these
compounds have been chosen for subsequent studies in MCF7
30 cells. DMSO (0.1%), representative of the highest concentration
used (100µg/ml), showed no effect on cell viability, confirming
its biological inertness.
Figure-1: Detection of apoptosis in MCF-7 cell line. (a) Induction of
apoptosis by 5b, 5c, 5i, 5g and 8. Cells were treated for 24 and 48 h with
indicated concentrations of the compounds and were co-stained with PI
65 and Annexin-V FITC prior to analysis using flow cytometry as described in
materials and methods.
The compounds induce the mitochondrial pathway of
apoptosis
Compounds 5(b, c, i, g) and 8 induce morphological changes
35 in MCF-7 cell
The mitochondrial membrane potential was measured by staining
It is desirable that cell death induced by an anticancer agent
70 with JC-1, a lipophilic cationic fluorescent dye capable of
selectively entering mitochondria and acting as a dual emission
probe that reversibly changes colour from red to green in concert
with polarization of mitochondrial membrane. Since oxygen
occurs through apoptosis. Light microscopic study of the cells
treated with IC50 concentrations of the compounds for 24 h
indeed revealed characteristic apoptotic changes like progressive
40 detachment and rounded cells. This study indicates that 5(b, c, i,
g) and 8 induced morphological changes in MCF-7 cells (see
supplementary fig S1).
consumption is
a
surrogate indicator of mitochondrial
75 bioenergetics, depleted cellular respiration coupled with
increased ROS production may lead to mitochondrial
2
| Journal Name, [year], [vol], 00–00
This journal is © The Royal Society of Chemistry [year]

Page 3 of 6
Medicinal Chemistry Communications
View Article Online
DOI: 10.1039/C4MD00469H
depolarization.27 A gradual time dependent change of red to green 30 known to function as the initial mediators of apoptosis.30To
fluorescence was observed in MCF7 cells after treatment with the
test samples (Figure 2). The green fluorescence in control cells
was 10.0%, which increased to 79.0 % for 5b, 62.3% for 5c, 50.0
for 5i, 46.1% for 5g, and 83% for 8 at 24 h of treatment and to
examine whether 5(b, c, i, g) and 8 affect the oxidative function
of the cell, we quantified ROS at different time points by
measuring the fluorescent signals of the products using FACS.
Cells were treated with IC50 concentrations of the samples for 24
5
88.9% for 5b, 89.8% for 5c, 97.5 for 5i, 93.1% for 5g, and 96.4 35 h (Figure 3a). The result showed significant accumulation of ROS
for 8 at 48 h. This was also reflected in the altered ratio of
green/blue fluorescence which in control cells was 9.0. Following
the addition of 5(b, c, i, g) and 8, this ratio demonstrated a time-
in this period. A shift to the right (red) generally indicates the
ROS accumulation. Figure 3a shows the percentage increase
caused by the compounds compared with control in different time
periods. The observed increase in fluorescence was attributable to
10 dependent decline (respectively 0.266, 0.605, 1.002, 1.173, and
0.205 at 24 h and 0.126, 0.114, 0.026, 0.069, and 0.037 at 48 h) 40 the ability of the compounds to generate ROS in MCF-7 cells.
suggesting the occurrence of depolarization of mitochondria by
Treatment of MCF-7 cells with test samples induced cleavage
of caspases and PARP
these compounds.
Caspases, a family of cysteine proteases, have emerged as key
45 enzymes in the regulation of apoptotic pathway. Activation of
caspases leads to the classical form of apoptotic pathway.31
Measurement of caspase activation was based on the ability of the
active caspase to cleave the chromophore from the enzyme
substrate. MCF-7 cells were treated with test samples for 24 h
50 and western blotting was performed for detection of caspase-9,
caspase-7 and PARP-1 cleavage products (Figure 3b). As shown
in Figure 3b, all the compounds induced the cleavage of initiator
caspase (caspase-9), effecter caspases (caspase-7) and PARP-1
enzyme. These results suggest that the compounds induce
55 apoptotic cell death pathway in the MCF-7 cells.
Figure-3: Detection of intracellular ROS generation and Caspases/PARP
cleavage assays. (a) MCF-7 cells (1×106 cells/ml) were treated with IC50
concentrations of 5b, 5c, 5i, 5g and 8 for 24 h. After treatment, cells
60 were washed and resuspended in PBS, incubated with CM-H2DCFDA for
30 min, and the fluorescence was measured using flow cytometry as
described in Materials and methods. Data is a representative of three
different experiments. (b) Cells were cultured in 6 cm dishes and treated
with IC50 concentrations of 5b, 5c, 5i, 5g and 8 for 48 h; cleavage of
65 caspases and PARP-1 was detected by Apoptosis Antibody Sampler Kit.
15 Figure-2: Detection of mitochondrial membrane potential. MCF-7 cells
were incubated with IC50 concentrations of 5b, 5c, 5i, 5g and 8 for 24 or
48 h. Cells were loaded with mitochondrial sensor dye JC-1 as described
in materials and methods. Change in membrane potential was detected
by the shift from red to green fluorescence in time dependent manner.
20
ROS induces DNA damage which plays role in apoptosis
induced by the compounds
The compounds induce S phase cell cycle arrest in MCF 7
cells
ROS plays an important role in the induction of apoptosis in
various types of cells. Mitochondria are the major site of ROS
-
25 production in mammalian cells, and superoxide (O2 ) appears to
Apoptosis occurs through cell cycle arrest at a specific phase of
70 cell division. The percentage increase or decrease in the number
of cells in each phase of the cell cycle compared to that in the
control was determined and represented by DNA histogram. As
summarised in Figure 4, treatment with apoptosis doses of these
be the primary ROS produced as the result of single electron
reduction of O2.28,29 However, excessive ROS generation beyond
a certain threshold level renders cancer cells highly susceptible to
chemotherapy induced death. Under such circumstances, ROS are
This journal is © The Royal Society of Chemistry [year]
Journal Name, [year], [vol], 00–00 | 3

Medicinal Chemistry Communications
Page 4 of 6
View Article Online
DOI: 10.1039/C4MD00469H
compounds for 24 and 48 h caused a gradual increase in the
number of cells in S phase. The increases in S and G2-M
populations at 24 h of treated MCF-7 cells (S and G2-M in Fig. 4)
were respectively 12.2%, 4.9% for 5b, 11.3%, 5.3% for 5c,
library of dispiro analogues containing both spiro-oxindole and
pyrrolidine/pyrrolizidine rings attached to andrographolide for
further biological evaluation. The findings indicate the promise
held out by N-benzyl glycine derivatives as five out of 12
5
14.8%, 2.0% (decrease) for 5i, 12.2%, 6.9% for 5g and 7.6%, 35 compounds show very promising activity against five cancer cell
5.6% for 8 in comparison with vehicle control. At 48 h, the
increase in S population was respectively 25.4% for 5b, 5c
19.5%, 5i 31.8%, 5g 26.7% and 22.3% for 8. In each case there
was a concomitant reduction in the number of cells in the G0-G1
lines. Of these, 5b, 5c, 5g and 5i carry CH3, Cl, F or Br group in
the aromatic ring. But the acenaphthoqinone ring containing
derivative (8) gave the best result.
We tested the effects of these compounds in five different cancer
10 (G0-G1 in Fig. 4) phase at 24 and 48 h in comparison with 40 cell lines (Figure 2A), viz. MCF7 (breast carcinoma), CHO
vehicle control (respectively 17.1%, 21.6% for 5b, 16.8%, 16.6%
for 5c, (12.9%, 26.2%) for 5i, (19.2%, 25.3%) for 5g and (13.2%,
27.2% for 8). This experiment revealed that the compounds
induced a dramatic increase in arrest cells at S phase at the two
15 time points.
(Chinese hamster ovary carcinoma), HepG-2 (hepatocellular
carcinoma), A431 (epidermoid carcinoma), and HeLa (cervical
carcinoma). Interestingly, all the compounds are relatively more
active against MCF-7 breast cancer cell line (Figure 2B). An
45 important feature of these lead derivatives is that cell death
triggered by them involves apoptosis. Typical features of
apoptosis like cell rounding, nuclear condensation and DNA
fragmentation, and externalization of phosphatidyl serine were
observed in response to treatment with 5(b, c, i, g) and 8.
50 Apoptosis induced by these derivatives were found to be
mediated via mitochondrial pathway with loss of mitochondrial
membrane potential, activation of caspases 9, cleavage of PARP
and increase of cytosolic cytochrome c level. They also induced
ROS mediated apoptotic cell death which was evidenced by flow
55 cytometric assay. The data suggests that the mode of action of the
derivatives on MCF-7 cells is similar to that of andrographolide.
Therefore these compounds may be regarded as potential
anticancer candidates.
From the above study it appears that three of this N-benzyl
60 glycine series of andrographolide derivatives (5c, 5g and 5i)
possess better activity compared to serine/proline series. It also
shows that chlorinated, fluorinated and brominated derivatives
are in general more potent than or iodinated derivatives, which
may be related to the electronegativity and size of the
65 substituents. But the only acenapthoquinone derivative (8) tested
appears most potent among all the derivatives.
Bromine is larger in size and also moderate halogen bond
acceptor than fluorine. The C-Br bond is stable enough, allowing
its insertion on diverse heterocycles of pharmacological value.
70 Another feature of brominated drugs concerns its binding affinity
for some cellular proteins. Replacing hydrogen by bromine also
provides a substantial alteration on the size and shape issues. The
subunits bearing bromine can be accommodated in tight and deep
cavities, as well as in hydrophobic pockets of the biological
75 targets.32 Chlorine occupy an intermediate position in between
fluorine and bromine. It is a better halogen bond acceptor, besides
being smaller in size than bromine. The C-Cl bond is also stable
enough, allowing its insertion on diverse heterocycles of
pharmacological value similar to bromine.
Figure-4: Study of cell cycle arrest in MCF-7 cells. DNA histograms show
the effect of treatment of 5b, 5c, 5i, 5g and 8 on cell-cycle of MCF-7 cells.
The cells were treated for 24 and 48 h with indicated concentrations of
20 the compounds. The area of peaks corresponds to DNA content in
different phases of cell cycle.
Discussion
80 From our study, three halogenated andrographolide derivatives
have been identified to show more potent cytotoxic activity than
andrographolide. Halogens are important functional groups. But
the number of halogenated anticancer drugs is less compared to
the derivatives having other functional groups. Binary protein
85 halogenated ligand complexes are more stable than non-
halogenated ligands because of favorable electrostatic
interactions of the halogen bonds. Cyclin-dependent kinase 2
(CDK2) plays critical roles in important intracellular pathways
In our previous studies, we evaluated the cytotoxic efficacy of 15
different di-spiropyrrolidino and dispiropyrrolizidino oxindole
25 andrographolide analogues prepared by cycloaddition of
azomethine ylide and sarcosine or proline at 12,13 double bond
of andrographolide. The tests performed using HCT116 and five
other carcinoma cell lines, viz. MiaPaCa-2, HepG2, HeLa, A375
and A549, revealed that three compounds showed better results
30 than andrographolide itself. The results encouraged to build a
4
| Journal Name, [year], [vol], 00–00
This journal is © The Royal Society of Chemistry [year]

Page 5 of 6
Medicinal Chemistry Communications
View Article Online
DOI: 10.1039/C4MD00469H
60 7. P. Siripong, B. Konckathip, K. Preechanukool, P. Picha, K.
Tunsuwan, W.C. Taylor, J. Sci. Soc. Thai., 1992, 18, 187–194.
8. A. Kapil, I. B. Koul, S.K. Banerjee, B.D. Gupta, Biochem.
Pharmacol., 1993, 46, 182–185.
and cell cycle progression, being thus an attractive biological
target for drug development.33-35 Auffinger and co-workers
showed that halogenated ligands bind to the cyclin-dependent
kinase 2 (CDK2), leading to more stable complexes than non-
halogenated ligands.36 The incorporation of halogen atoms into
new bioactive chemical entity is commonly used to increase
membrane permeability and thereby improve oral absorption.37
Halogenation also enhances the blood brain barrier (BBB)
permeability and this is a pre-requisite for many drugs.38 In this
9. R.J. Srinivasa, J. Stanslas, N.H. Lajis, S. Said, A.S. Hamzah, A.
5
65
70
75
McCarroll, C. Matthews, M. F. G. Stevens, Brit. J. Cancer, 2003,
88, S25–S54.
10. S. Rajagopal, R. A. Kumar, D. S. Deevi, C. Satyanarayana, R.
Rajagopalan, J. Exp. Ther. Oncol., 2003, 3, 147-158.
11. C.-G. Jiang, J.-B. Li, F.-R. Liu, T. Wu, M. Yu, H.-M. Xu,
Anticancer Res., 2007, 27, 2439-2447.
10 respect our study of new halogenated anticancer agents are
important findings and adds a new approach in cancer research. It
is believed that carbon-halogen bonds are not easily metabolized
by the cytochrome p450 system and, therefore, it is a feasible
strategy to block the metabolically labile positions of a particular
15 new bioactive chemical entity,39 providing an explanation for our
results.
12. M. Geethangili, Y. K. Rao, S.- H. Fang, Y.-M. Tzeng, Phytother.
Res., 2008, 22, 1336-1341.
13. (a) L. Yang, D. Wu, K. Luo, S. Wu, P. Wu, Cancer Lett., 2009, 276,
180-188. (b) J. Li, H.-Y. Cheung, Z. Zhang, G. K. L. Chan, W.-F.
Fong, Eur. J. Pharmacol., 2007, 568, 31-44.
14. S. Nanduri, V. K. Nyavanandi, S. S. R. Thunuguntla, S. Kasu, M. K.
Pallerla, P. S. Ram, S. Rajagopal, R. A. Kumar, R. Ramanujam, J.
M. Babu, K. Vyas, A. S. Devi, G. O. Reddy, V. Akella, Bioorg.
Med. Chem. Lett,. 2004, 14, 4711-4717.
Conclusion
80 15. S. R. Jada, C. Matthews, M. S. Saad, A. S. Hamzah, N. H. Lajis, M.
F. G. Stevens, J. Stanslas, Br. J. Pharmacol., 2008, 155, 641-654.
16. S. R. Jada, G. S. Subur, C. Matthews, A. S. Hamzah, N. H. Lajis, M.
S. Saad, M. F. G. Stevens, J. Stanslas, Phytochemistry, 2007, 68,
904-912.
In conclusion, our study demonstrates that five andrographolide
derivatives show more potent anticancer activity than
20 andrographolide itself. In this respect, our findings are important
to explore anticancer agent(s). Probably, biological potency of the
derivatives results from the combined effect of the nature of
halogens and the structures of the compounds to which they are
attached. In addition, we can conclude that the potent compound
25 8, which is structurally different from other analogues to some
extent, may be similar to andrographolide in its effect towards
apoptosis induction in cancer cells.
85 17. C. Satyanarayana, D. S. Deevi, R. Rajagopalan, N. Srinivas, S.
Rajagopal, BMC Cancer, 2004, 4, 26. 1-8.
18. H.- W. Xu, J. Zhang, H.-M. Liu, J.-F. Wang, Synth. Commun. 2006,
36, 407-414.
19. A. Hazra, P. Paira, K. B. Sahu, S. Naskar, P. Saha, R. Paira, S.
90
Mondal, A. Maity, P. Luger, M. Weber, N. B. Mondal, S. Banerjee,
Tetrahedron Lett., 2010, 51, 1585−1588.
20. S. K. Dey, D. Bose, A. Hazra, S. Naskar, A. Nandy, R. N. Munda,
S. Das, N. Chatterjee, N. B. Mondal, S. Banerjee, K. Das Saha,
PLoS ONE, 2013, 8(3), e58055.
Acknowledgement
30 The authors express their gratitude to the Director, IICB for
laboratory facilities, and the Indian Council for Medical Research
(ICMR-59/03/2007/BMS/TRM) and WB-DST for providing the
funding and fellowship to DC. NBM is indebted to CSIR for the
award of Emeritus Scientist scheme. We express our gratitude to
35 Dr. Sukdeb Banerjee and Dr. B. Achari (Ex. Emeritus Scientist,
CSIR) for their valuable suggestions. CKJ was supported by
UGC-senior research fellowship.
95 21. A. Hazra, Y. P. Bharitkar, D. Chakraborty, K S. Mondal, N. Singal,
S. Mondal, A. Maity, R. Paira, S. Banerjee, N. B. Mondal ACS
Comb. Sci. 2013, 15, 41−48.
22. G. Makin, J. A. Hickman, Cell. Tissue. Res., 2000, 301(1), 143–152.
23. H. Jyonouchi, S. Sun, K. Iijima, M. Wang, S. S. Hecht,
100
Carcinogenesis, 1999, 20(1), 139–145.
24. M. K. N. Gorter, Recl. Trav. Chim. Pays-Bas., 1911, 30, 151-160.
25. A. B. Smith, B. H. Toder, P. J. Carrol, J. Donohue, Crystallogr.
Spectr. Res., 1982, 12, 309-319.
Conflict of interest
26. M. Rajani, N. Shrivastava, M. N. Ravishankara, Pharm. Biol.,
2000, 38(3), 204-209.
40 None
105
27. N. Jr. Hail, P. Chen, L. R. Bushman, Neoplasia, 2010, 12, 464–475.
28. A. J. Lambert, M. D. Brand, Biochem. J., 2004, 382, 511–517.
29. T. M. Paravicini, C. G. Sobey, Clin. Exp. Pharmacol. Physiol.,
2003, 30 (11), 855–859.
Notes and Reference
aDepartment of Chemistry, bMolecular Parasitology Laboratory, cCancer
Biology and Infllamatory Disorder Division, Indian Institute of Chemical
45 Biology, Council of Scientific and Industrial Research, 4 Raja S C
Mullick Road, Jadavpur, Kolkata 700032, West Bengal, India.
dDepartment of Pharmaceutical Technology, Division of Medicinal and
Pharmaceutical Chemistry, PO Box No. 17020, Jadavpur University,
Kolkata-700 032, West Bengal, India.
110 30. L. Gibellini, M. Pinti, M. Nasi, S. D. Biasi, E. Roat, Cancers, 2010,
2, 1288–1311.
31. M. Leist, M. Jaattela, Nat. Rev. Mol. Cell. Biol., 2001, 2, 589–598.
32. G. Siegal, E. Ab, J. Schultz, Drug Discovery Today, 2007, 12,
1032–1039.
50 † Electronic Supplementary Information (ESI) available: [details of any
supplementary information available should be included here]. See
DOI: 10.1039/b000000x/
115 33. W. F. Jr. De Azevedo, S. Leclerc, L. Meijer, L. Havlicek, M. Strnad,
Eur. J. Biochem. 1997, 243, 518–526.
1. Traditional medicine strategy launched. WHO News, 2002, 80, 610.
34. W. F. Jr. De Azevedo, F. Canduri, N. J. da Silveira, Biochem.
Biophys. Res. Commun. 2002, 293, 566–571.
2. S. Benowitz, Scientist 1996, 10, 1−7.
55 3. D. J. Newman, G. M. Cragg, J. Nat. Prod., 2007, 70, 461−477.
4. http://www.who.int/healthinfo/global_burden_disease/en/
5. http://www.iarc.fr/en/media-
35. W. F. Jr. De Azevedo, R. T. Gaspar, F. Canduri, J. C. Jr. Camera, N.
120
J. F. Silveira, Biochem. Biophys. Res. Commun., 2002, 297, 1154–
1158.
centre/iarcnews/2010/globocan2008.php.
36. P. Auffinger, F. A. Hays, E. Westhof, P. S. Ho, Proc. Natl. Acad.
Sci., USA 2004, 101, 16789–16794.
6. S.S. Handa, A. Sharma, Indian J. Med. Res., 1990, 92, 284–292.
This journal is © The Royal Society of Chemistry [year]
Journal Name, [year], [vol], 00–00 | 5

Medicinal Chemistry Communications
Page 6 of 6
View Article Online
DOI: 10.1039/C4MD00469H
37. G. Gerebtzoff, X. Li-Blatter, H. Fischer, A. Frentzel, A. Seelig,
Chem. Bio. Chem., 2004, 5, 676–684.
38. C. L. Gentry, R. D. Egleton, T. Gillespie, T. J. Abbruscato, H. B.
Bechowski, Peptides 1999, 20, 1229–1238.
5
39. M. Z. Hernandes, S. M. Cavalcanti, D. R. Moreira, W. F. Jr. de
Azevedo, A. C. Leite, Curr. Drug Targets, 2010, 11(3), 303–314.
6
| Journal Name, [year], [vol], 00–00
This journal is © The Royal Society of Chemistry [year]