Synthesis and biological evaluation of novel hybrid compounds between chalcone and piperazine as potential antitumor agents

Article abstract of DOI:10.1039/c5ra20197g

Chalcones play an important role in living organisms with a wide range of biological activities including potent antitumor activity. Previously, we reported that N-aryl piperazine compounds have excellent biological activity. To further explore the struct

Full text of DOI:10.1039/c5ra20197g

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Synthesis and biological evaluation of novel hybrid compounds
between chalcone and piperazine as potential antitumor agents†
Received 00th January 20xx,
Accepted 00th January 20xx
Zewei Maoa, Xi Zhengb, Yan Qib, Mengdi Zhanga, Yao Huanga, Chunping Wan*b and Gaoxiong Rao*a
Chalcones play an important role in living organisms with a wide range of biological activities including
potent antitumor activity. Previously, we reported that Nꢀaryl piperazine compounds have excellent biological
activity. To further explore the structureꢀactivity relationships, a series of novel hybrid compounds between
chalcone and piperazine have been synthesized, and their in vitro antitumor activity was evaluated against a
panel of human tumor cell lines. The results demonstrated that compounds bearing acetophenone showed
better anticancer activity than cisplatin and other hybrid compounds, and that substitution of the acetophenone
with halogen atom, was vital for modulating cytotoxic activity. Among all synthetic derivatives, hybrid
compound 7c was found to be the most potent compound against A549, Hela and SGC7901 (IC50=5.24ꢁM,
0.19ꢁM and 0.41ꢁM, respectively) , importantly, 7c exerted obvious inhibitory effect in vivo.
DOI: 10.1039/x0xx00000x
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HO
O
NH
N
N
N
Introduction
O
N
N
OH
O
HO
F
O
Chalcones and their derivatives are a class of important
organic compounds that play an important role in living
organisms1 with broad range of biological activities, including
antitumor,2 antioxidant,3antiꢀinflammatory,4 and so on. Chalcones
are widely present in nature, and a number of chalcones have been
reported in recent years (Fig.1).5ꢀ8Present studies show that the
antitumor activity of chalcones may result from multiple
mechanisms, including inhibition of proliferation and promotion of
apoptosis of tumor cells, inhibition of tyrosine kinases, as well as
selective cytotoxic activity, which suggest these molecules may be
used as potential antitumor agents and have attracted more
pharmaceutical chemists into the exploration of novel chalcone
antitumor drugs.9ꢀ12
Levodropropizine
Eperezolid
O
O
F
COOH
R
NH
N
N
N
O
N-4-Piperazinyl-Ciprofloxacin-Chalcone Hybrids
Fig.2 Structures of Nꢀaryl piperazine agents.
Molecular hybridization has been one of the successfully applied
strategies for the design and synthesis of novel and efficient
biological agents. Molecular hybridization involves the combination
of two distinct pharmacophores to form new hybrid molecules. For
example, Nꢀ4ꢀpiperazinylꢀciprofloxacinꢀchalcone hybrids have been
found to exhibit a broadꢀspectrum of antitumoractivities.15Moreover,
in previous research, we have reported the synthesis of a series of
novel Nꢀaryl piperazine hybrid compounds and their potential
antitumor activity.16ꢀ17In the present research, we have designed and
synthesized new molecules towards the recombination of chalcone
and piperazine in the same structure through medicinal chemistry
hybridization (Fig.3). The potential cytotoxicity of derivatives was
evaluated in vitro against a panel of human tumor cell lines and in
Similarly, Nꢀaryl piperazine moieties represent
a series of
important organic compounds that make up the core structures in
medicines, such as Eperezolid13and Levodropropizine14. Recently,
Nꢀaryl piperazine derivatives have attracted considerable interests
for their versatile properties in chemistry and pharmacology, and
these moieties have been developed to drug molecular design
widely.
OH
O
HO
O
OH
OH
O
OH
OH
O
Sappanchalcone
4-hydroxyderricin
vivo.
O
HO
OH
O
R
R
N
hybridization
O
O
O
N
Dimethylaminochalcone
Licochalcone A
and derivatization
N
R
Fig. 1 Structures of natural chalcones with antitumor activities.
R
N
R
N
R
Fig. 3 Designed strategy of chalcone hybrids.
a.School of Traditional Chinese Medicine, Yunnan University of Traditional Chinese
Medicine, Kunming, 650500, P. R. China. E-mail:rao13987124569@qq.com.
b.Central laboratory, The NO.1 Affiliated Hospital of Yunnan University of
Traditional Chinese Medicine, Kunming, 650021, P. R. China. E-mail:
wanchunping1012@163.com.
†Electronic Supplementary Informaꢀon (ESI) available: Synthesis and
characterization of the compounds. See DOI: 10.1039/x0xx00000x
This journal is © The Royal Society of Chemistry 20xx
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O
O
Table 1. Structures of title compounds.
Compound R or X
a
O
b
m. p(°C)
Yield (%)a
N
N
F
F
5a
5b
H
CH3
231ꢀ233
231ꢀ234
87
91
3
2
1
O
O
5c
C2H5O
235ꢀ236
86
c or d
f
N
N
N
N
5d
F3C
240ꢀ242
83
NH
R
N
4
5a-5h
O
O
5e
5f
236ꢀ238
247ꢀ249
84
78
Cl
Cl
Cl
e
Cl
O
O
N
N
N
O
R
7a-7d
N
O
Cl
N
N
S
5g
5h
239ꢀ241
237ꢀ239
82
70
R
O
6a-6f
F
Scheme 1 Synthesis of hybrid derivatives. Reagents and conditions: (a) 20%
KOH, EtOH, rt, 24h; (b) piperazine hexahydrate, Cs2CO3, DMF, 110°C, 12h;
(c) RCOCl, Et3N, DCM, 0°C, 2h; (d) RCOOH, DCC, DMAP, DCM, 24h; (e)
RSO2Cl, Pyridine, DCM, rt, 12h; (f) Cs2CO3, 2ꢀbromoacetophenone, DCM, rt,
4h.
Br
6a
6b
6c
234ꢀ236
238ꢀ239
240ꢀ241
78
85
84
H3C
F
Results and discussion
Br
F
Chemistry
The general route used for the synthesis of title compounds is
shown in Scheme 1. Treatment of commercially available 4ꢀ
fluoroacetophenone (1) with 4ꢀdimethylaminobenzaldehyde
(2)gave the 4ꢀdimethylaminoꢀ4’ꢀfluorochalcone (3) in the
presence of KOH resulted by the Aldol condensation.
Subsequently, the key intermediate (4) was prepared by
substitution of fluorine atom of3 with piperazine following the
literature procedure.18Although in the product4 preparation from
chalcone intermediate (3) by nucleophilic substitution reaction
with Nꢀheterocycles, there is a possibility for AzaꢀMichael
addition product,19ꢀ20 no Michaelꢀaddition products were
observed in our work.
Finally, acylation and sulfonylation of NH group with acyl
chloride or carboxylic acid and sulfonyl chloride afforded hybrid
compounds 5aꢀ5h and 6aꢀ6f in good yields (62ꢀ91%),
respectively. For further insight toward the structure and activity
relationship, a few tertiary amines (7aꢀ7d) were synthesized by
treatment of 2ꢀbromoacetophenone as well. Structures of novel
hybrid compounds are shown in Tables 1.
6d
240ꢀ242
86
Br
6e
6f
242ꢀ243
238ꢀ240
250ꢀ252
252ꢀ254
251ꢀ252
254ꢀ256
71
62
86
84
85
79
F3C
O2N
7a
7b
7c
7d
F
Cl
Br
a Yields represent isolated yields.
for hybrid 6e against A549.It could be seen that chloro or
trifluoromethyl substituent of the benzene ring might be
attributed to the cytotoxic activity for amides and sulfonamides.
To our delight, hybrid compounds bearing acetophenone all
showed a higher potency anticancer activity than cisplatin and the
above hybrid compounds. Interestingly, the halogen atom
substituent atpositionꢀ4 of the benzene ring was more sensitive to
cytotoxic activity. Notably, derivatives 7c displayed the best IC50
value against 3 tumor cell lines. In comparison to compounds 7b-
7d, compound 7a was less potent against all tumor cell lines.
Derivatives 7c showed best antiꢀtumoral activity in vitro as
described above. In order to further evaluate in vivo antiꢀtumoral
effect of 7c, S180 tumorꢀbearing mice was established for
detecting its in vivo antiꢀtumoral effect. The result indicated that
daily administration of 25mg/kg 7c significantly inhibited the
growth of tumor cell, implying 7c may be a potential therapeutic
agent for tumor (Fig. 4).
Biological assay
Cytotoxic activity of novel synthesized hybrid derivatives were
evaluated against human lung cancer cell line A549, human cervical
carcinoma Hela and human gastric carcinoma SGC7901 by MTT
assay,21 using cisplatin as the reference drug. The biological results
of hybrid compounds are summarized in Tables 2.
As shown in Table 2, the structures of the hybrid compounds
have an obvious influence on antitumor activities. Amide
derivatives were more active in general, and the dichloro or
trifluoromethyl substituent of the benzene ring displayed similar
or more cytotoxic activity in vitro compared to cisplatin.
Especially, compound 5d showed similar antitumor activity
against Hela and SGC7901 cells, similarly, compounds 5f and 5g
displayed potent or similar cytotoxic activity against A549 and
SGC7901 cells.
The results suggest that the existence of halogen atom has
played an important role in the antitumor activity of hybrids.
Generally, the chlorine atom substituent atpositionꢀ4 of benzene
ring, as well as the fluorine group exhibit higher cytotoxic
activity. The structureꢀactivity relationship (SAR) results were
summarized in Scheme 2.
However, sulfonamide compounds were inactive against 3
tumor cell lines investigated at the concentration of 40ꢁMexcept
2 | J. Name., 2012, 00, 1-3
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O
Table 2. In vitro cytotoxic activities of hybrid compounds.
O
Cell lines (IC50, ꢁM) a, b
O
Compound
N
>
>
N
A549
>40
>40
>40
>40
Hela
>40
>40
>40
16.32±2.02
>40
SGC7901
>40
>40
>40
17.29±1.25
>40
14.16±2.68
N
R
N
R
N
N
5a
5b
5c
5d
5e
5f
O
(A)
(B)
Better
O
N
O
S R
O
>40
N
N
10.36±2.35
>40
(C)
15.13±2.97
>40
>40
>40
>40
>40
16.59±2.73
35.76±4.13
8.72±2.10
>40
>40
>40
>40
>40
34.18±3.71
5g
5h
6a
6b
6c
6d
6e
>40
>40
>40
>40
>40
>40
>
F
>
Cl
>
H
R (A) =
Br
7c
the best
7a
7b
7d
Best Potent Compound
O
>40
>40
>40
6f
7a
7b
7c
7d
cisplatin
O
N
11.12±2.13
7.17±0.88
5.24±1.01
9.43±1.27
11.54±1.15
2.08±0.75
1.02±0.49
0.19±0.13
0.26±0.18
20.52±1.84
8.84±1.59
0.16±0.11
0.41±0.26
4.04±1.03
12.44±2.12
N
N
Cl
7c
Scheme 2 Structureꢀactivity relationship of hybrid compounds.
a Cytotoxicity as IC50 values for each cell line, the concentration of compound
that inhibit 50% of the cell growth measured by MTT assay.
b Each value was reproduced in triplicate.
Experimental
General procedure for the preparation of chalcone derivatives
5a-5h.
On the whole, although compounds 7aꢀ7d werefound toexhibit
broadꢀspectrum of antitumor activities, they also showed no
pronounced selectivity. Amide hybrids displayed similar active
and better selectivity compared to 7aꢀ7d. These hybrids were to
be lead compounds for further structural modification and
activity research.
Method A: To a stirred solution of compound4 (0.5mmol) and
pyridine (0.5mL) in dried DCM (10mL), acylchloride (1.0mmol)
was added and reaction mixture was stirred for 2 h at 0°C. After
completion of the reaction as indicated by TLC, the reaction was
quenched by the addition of saturated NaHCO3 (20mL) and was
extracted with CHCl3 (3×10mL). The organic layer was dried using
anhydrous sodium sulfate, concentrated in vacuo and purified by
column chromatography (DCM) to afford products.
Conclusions
Method B: To a stirred solution of DCC (1.0mmol) and carboxylic
acid(1.0mmol)in dried DCM (10mL), compound4(0.5mmol) was
added and reaction mixture was stirred for 24h at rt. After
completion of the reaction as indicated by TLC, the mixture was
filted and the filtrate was concentrated in vacuo and purified by
column chromatography (DCM) to afford compounds.
In summary, a series of novel hybrid compounds between
chalcone and piperazine have been synthesized and proved to be
potent antitumor agents. The chalcone hybrids 7a-7d bearing
acetophenone were found to be the most potent compounds, and
halogen atom substituent atpositionꢀ4 of benzene ring were vital
for modulating cytotoxic activity. Among all synthetic derivatives,
hybrid compound 7c was found to be the most potent compound in
treatment of tumor in vitro and in vivo. Further molecular
mechanism is currently underway and the results will be reported
in due course.
1
Compound 5d:Yellow solid; H NMR (300MHz, CDCl3) δ: 7.99(d,
J=8.7Hz, 2H), 7.68ꢀ7.79 (m, 3H), 7.54ꢀ7.60(m, 2H), 7.53(d,
J=8.7Hz, 2H), 7.37(d, J=15.3Hz, 1H), 6.90(d, J=8.7Hz, 2H), 6.67(d,
J=8.7Hz, 2H), 3.90(s, 2H), 3.57(s, 2H), 3.35(s, 4H), 2.99(s, 6H); 13
C
NMR (75MHz, CDCl3) δ: 188.2, 168.8, 153.2, 151.8, 144.4, 136.2,
130.4, 130.3, 130.2, 130.1, 129.2, 126.8, 124.2, 122.8, 116.5, 114.27,
111.8, 47.7, 40.1.HRMS: m/z calcd for C29H29F3N3O2
(M+H)+508.2206, found 508.2204.
1
Compound 5f: Brown solid; H NMR (300MHz, CDCl3)δ: 8.00(d,
J=8.7Hz, 2H), 7.79(d, J=15.3Hz, 1H), 7.55(d, J=8.7Hz, 2H), 7.38(s,
1H), 7.38(d, J=15.3Hz, 1H), 7.16ꢀ7.20(dd, J=2.7Hz, 2.4Hz, 1H),
6.98(d, J=8.7Hz, 1H), 6.91(d, J=9.0Hz, 2H), 6.70(d, J=9.0Hz, 2H),
4.80(s, 2H), 3.75ꢀ3.81(m, 4H), 3.31ꢀ3.37(m, 4H), 3.03(s, 6H); 13C
NMR (75MHz, CDCl3)δ: 188.5, 165.8, 153.3, 152.1, 152.0, 144.7,
130.5, 130.3, 128.0, 127.1, 123.7, 123.1, 116.8, 114.4, 114.4, 112.0,
68.9, 40.3, 34.1, 25.1; HRMS: m/z calcd for C29H30Cl2N3O3
(M+H)+538.1659, found 538.1653.
Fig. 4 In vivo antiꢀtumoral effect of 7c in S180 tumorꢀbearing mice. Female
BALB/c mice was injected subcutaneously into the right forelimb armpit
with 1×106 S180 tumor cells, 24 h after injected tumor, 7c or vehicle was
Compound 5g: Pale yellow solid; 1H NMR (300MHz, CDCl3)δ:
8.01(d, J=8.7Hz, 2H), 7.80(d, J=15.3Hz, 1H), 7.50ꢀ7.56(m, 4H),
administered via intragastric administration, the experiment was terminated 7.38(d, J=15.3Hz, 1H), 7.26ꢀ7.30(dd, J=2.1Hz, 2.4Hz, 1H), 6.93(d,
after 21 days and the mice were sacrificed, tumors were harvest and weighed.
Results presented are mean± s.e.m, n = 8. *P<0.05, **P<0.01 versus vehicle
group.
J= 9.0Hz, 2H), 6.70(d, J=8.7Hz, 2H), 3.87(s, 2H), 3.63(s, 2H),
3.37(s, 4H), 3.03(s, 6H); 13C NMR (75MHz, CDCl3)
δ: 188.4, 168.1,
153.3, 152.0, 144.7, 135.2, 134.5, 133.2, 130.8, 130.4, 130.3, 129.4,
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126.6, 123.0, 116.7, 114.4, 111.9, 40.2, 34.0, 25.0; HRMS: m/z calcd DMSO was added to each well and the plate was agitated at
for C28H28Cl2N3O2 (M+H)+508.1553, found 508.1550.
oscillator for 5 min to dissolve the MTTꢀformazan. The assay plate
General procedure for the preparation of chalcone derivatives was read at a wavelength of 570 nm using a microplate reader.
6a-6f: To a stirred solution of compound4 (0.5mmol) and pyridine Anti-tumoral effect of 7c in vivo: Female BALB/c mice was
(0.5mL) in dried DCM (10mL), sulfonyl chloride (1.0mmol) was injected subcutaneously into the right forelimb armpit with 1×106
added and reaction mixture was stirred for 12h at room temperature. S180 tumor cells that resuspended in 0.2 ml of PBS, 24h after
After completion of the reaction as indicated by TLC, the reaction inoculated, the mouse were then randomly divided into two groups:
was quenched by the addition of 10%NaOH(20mL) and was a vehicle control group, and 7c group. Dosage of 7c group was orally
extracted with CHCl3 (3×10mL). The organic layer was dried using administered 25mg/kg 7c.21 days after the last drug administration,
anhydrous sodium sulfate, concentrated in vacuo and purified by the mice were sacrificed, tumors were harvest and weighed. Data are
column chromatography (DCM) to afford products.
expressed as mean± s.e.m. of indicated experiments. Student’s t test
1
Compound 6e: Yellow solid; H NMR (300MHz, CDCl3)δ: 7.97(d, was used to determine significance between two groups where
J=8.7Hz, 2H), 7.94(d, J=9.0Hz, 2H), 74ꢀ7.84(m, 3H), 7.54(d, appropriate. P<0.05 was considered to be statistically significant.
J=8.4Hz, 2H), 7.35(d, J=15.6Hz, 1H), 6.88(d, J=8.4Hz, 2H), 6.70(d,
J=8.4Hz, 2H), 3.43(d, J=5.1Hz, 4H), 3.21(s, 4H), 3.03(s, 6H);
HRMS: m/z calcd for C28H29F3N3O3S (M+H)+544.1876, found
544.1875.
General procedure for the preparation of chalcone derivatives
Acknowledgements
This work was financially supported by the Natural Science
Foundation of China (81460624) and the Application Basic Research
7a-7d. To a stirred solution of compound4 (0.5mmol) and Cs2CO3
Program of Yunnan Province (2013FZ088, 2014FZ087). We
(1.0g) in dried DCM (10mL), 2ꢀbromoacetophenone (0.6mmol) was
acknowledge Prof. Hongbin Zhang (Yunnan University, China) for
added and reaction mixture was stirred for 4h at rt. After completion
the NMR analysis. We are grateful to Miss Tiffany. Liao (Boston
of the reaction as indicated by TLC, the reaction was quenched by
Children’s Hospital, USA) for her useful help on English quality.
the addition of 10% NaOH(20mL) and was extracted with CHCl3
(3×10mL). The organic layer was dried using anhydrous sodium
sulfate, concentrated in vacuo and purified by column
chromatography (1% Et3N/DCM) to afford products.
Compound 7a: Yellow solid; H NMR (300MHz, CDCl3)δ: 7.99(d,
Notes and references
1
1
2
3
4
F.Berrue, M. W. B. McCulloch, R. G. Kerr. Bioorg. Med.
Chem., 2011, 19, 6702ꢀ6719.
L. Jayasinghe, B. A. I. S. Balasooriya, W. C. Padmini, N.
Hara, Y. Fujimoto. Phytochemistry., 2004, 65, 1287ꢀ1290.
K. Sugamoto, Y. I. Matsusita, K. Matsui, C. Kurogi, T.
Matsui. Tetrahedron., 2011, 67, 5346ꢀ5359.
N. Mishra, P. Arora, B. Kumar, L. C. Mishra, A.
Bhattacharya, S. K. Awasthi,V. K. Bhasin.Eur. J. Med.
Chem., 2008, 43, 1530ꢀ1535.
J=8.1Hz, 4H), 7.79(d, J=15.3Hz, 1H), 7.50ꢀ7.54(m, 3H), 7.46(d,
J=7.5Hz, 2H), 7.39(d, J=15.6Hz, 1H), 6.89(d, J=8.7Hz, 2H), 6.66(d,
J=8.7Hz, 2H), 3.84(s, 2H), 3.41(t, J=4.2Hz, 4H), 2.97(s, 6H), 2.74(t,
J=4.5Hz, 4H); 13C NMR (75 MHz, CDCl3)δ: 196.1, 188.2, 153.7,
151.7, 144.1, 135.9, 133.4, 130.3, 130.1, 129.1, 128.6, 128.1, 123.0,
116.7, 113.6, 111.8, 64.2, 53.1, 47.3, 40.1; HRMS: m/z calcd for
C29H32N3O2 (M+H)+454.2489, found 454.2488.
Compound 7b: Yellow solid; 1H NMR (300MHz, CDCl3)δ: 8.02ꢀ
8.07(dd, J=5.7Hz, 5.4Hz, 2H), 7.99(d, J= 8.7Hz, 2H), 7.79(d,
J=15.6Hz, 1H), 7.54(d, J=8.7Hz, 2H), 7.38(d, J=15.3Hz, 1H), 7.14(t,
J=8.4Hz, 2H), 6.90(d, J=8.7Hz, 2H), 6.68(d, J=8.7Hz, 2H), 3.81(s,
5
6
7
G. S. Jeong, D. S. Lee, B. Li, H. J. Lee, E. C. Kim, Y. C.
Kim.Eur. J. pharmacol., 2010, 644, 1ꢀ3.
T. Akihisa, T. Kikuchi, H. Nagai, K. Ishii, K. Tabata, T.
Suzuki.J Oleo Sci., 2011, 60, 71ꢀ77.
M. Chen, S. B. Christensen, J. Blom, E. Lemmich, L.
Nadelmann, K. Fich, T. G. Theander, A. Kharazmi.
Antimicrob Agents Chemother., 1993, 37, 2550ꢀ2556.
C. Franco, F. Rossella, B. Adriana, C. Paola, O. S. Daniela,
R. Francesca, Y. Matilde, Francisco, O. Francesco, A.
Stefano. J. Med. Chem., 2009, 52, 2818ꢀ2824.
2H), 3.42(t, J=4.2Hz, 4H), 3.00(s, 6H), 2.74(t, J=4.8Hz, 4H); 13
C
NMR (75 MHz, CDCl3)δ: 194.8, 188.4, 164.2, 153.8, 151.9, 144.2,
132.4, 131.1, 131.0, 130.4, 130.2, 129.3, 123.1, 116.9, 115.0, 116.66,
113.8, 111.9, 64.5, 53.2, 47.4, 40.2; HRMS: m/z calcd for
C29H31FN3O2 (M+H)+472.2395, found 472.2393.
8
9
Compound 7c: Yellow solid; 1H NMR (300MHz, CDCl3)δ: 7.99(t,
J=7.2Hz, 4H), 7.79(d, J=15.3Hz, 1H), 7.54(d, J=8.4Hz, 2H), 7.44(d,
J=8.4Hz, 2H), 7.39(d, J=15.6Hz, 1H), 6.91(d, J=9.0Hz, 2H),, 6.69(d,
J=8.7Hz, 2H), 3.82(s, 2H), 3.42(t, J=4.5Hz, 4H), 3.01(s, 6H), 2.75(t,
J=4.8Hz, 4H); 13C NMR (75 MHz, CDCl3)δ: 195.2, 188.4, 153.8,
151.9, 144.3, 139.9, 134.2, 130.4, 130.2, 129.8, 129.4, 129.0, 123.1,
116.9, 113.8, 111.9, 64.5, 53.2, 47.4, 40.2; HRMS: m/z calcd for
C29H31ClN3O2 (M+H)+488.2099, found 488.2100.
J. Chen, J. Yan, J. H.Hu, Y. Q. Pang, L. Huang, X.S. Li. RSC
Adv., 2015, 5, 68128ꢀ68135.
10 Y. N. Shen, L. Lin, H.Y. Qiu, W. Y. Zou, Y. Qian, H. L. Zhu.
RSC Adv., 2015, 5, 23767ꢀ23777
11 K. V. Sashidhara, M. Kumar, R. K. Modukuri, R.Sonkar, G.
Bhatia, A. K. Khanna, S.Rai, R. Shukla. Bioorg. Med. Chem.
Lett.,2011,21, 4480ꢀ4484.
12 C. Tang, L. H. Zhu, Y. Chen, R. Qin, Z. N Mei, J. Xu, G. Z.
Yang. RSC Adv., 2014, 4, 10862ꢀ10874.
1
Compound 7d: Yellow solid; H NMR (300MHz, CDCl3)δ: 7.99(d,
J=8.7Hz, 2H), 7.88(d, J=8.7Hz, 2H), 7.79(d, J=15.3Hz, 1H), 7.59(d,
J=8.4Hz, 2H), 7.54(d, J=8.7Hz, 2H), 7.39(d, J=15.6Hz, 1H), 6.90(d,
J=9.0Hz, 2H), 6.68(d, J=8.7Hz, 2H), 3.80(s, 2H), 3.41(t, J=4.8Hz,
4H), 3.00(s, 6H), 2.73(t, J=4.8Hz, 4H); 13C NMR (75 MHz, CDCl3)δ:
195.3, 188.3, 153.7, 151.8, 144.2, 134.6, 132.0, 130.4, 130.2, 129.8,
129.3, 128.6, 123.0, 116.8, 113.8, 111.9, 64.4, 53.1, 47.4, 40.2;
HRMS: m/z calcd for C29H31BrN3O2 (M+H)+532.1594, found
532.1592.
13 C.Edlund, H. Oh, C. E.Nord. ClinMicrobiolInfec., 1999, 1,
51ꢀ53.
14 G. Fasciolo, A. Nicolini, N. Vacca, P. Viglierchio. Current
Therapeutic Research., 1994, 3, 251ꢀ261.
15 M. AbdelꢀAziz, S. E. Park, E. D. Gamal, M. A. AbuoꢀRahma,
Y. K. Sayed. Eur. J. Med. Chem., 2013, 69, 427ꢀ438.
16 Z. W. Mao, C. P. Wan, X. Zheng, Y. Jiang, G.X. Rao.
China Pharm Univ., 2014, 5, 517ꢀ521.
17 Z. W. Mao, C. P. Wan, Y. Jiang, W. L. Guo, G.X. Rao.
China Pharm Univ., 2015, 1, 58ꢀ61.
J
J
Antitumor activity: About 1×104 cell/well were seeded into 96ꢀwell
microtiter plates. After twentyꢀfour hours postꢀseeding, cells were
treated with vehicle control or various concentrations of samples for
48h.20ꢁL of MTT solution (5mg/mL) was added to each well and
the tumor cells were incubated at 37°C in a humidified atmosphere
of 5% CO2 air for 4 h. Upon removal of MTT/medium, 150ꢁL of
18 A. Ajay, V. Singh, S. Singh, S. Pandey, S. Gunjan, D. Dubey,
S. K. Sinha, B. N. Singh, V. Chaturvedi, R. Tripathi, R.
4 | J. Name., 2012, 00, 1-3
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DOI: 10.1039/C5RA20197G
Journal Name
ARTICLE
Ramchandran, R. P. Tripathi. Bioorg. Med. Chem., 2010, 23,
8289ꢀ8301.
19 A. J. Watson, A. J. Fairbanks. Eur. J. Org. Chem., 2013, 30,
6784ꢀ6788.
20 M. L. Kantam, J. Yadav, S. Laha, B. Sreedhar, S. Jha. Adv.
Synth. Catal., 2007, 349, 1938ꢀ1942.
21 Z. W. Mao, Y. Li, J. B. Chen, Y. Y. Wang, H. B. Zhang.
Bioorg. Med. Chem. Lett., 2010, 20, 4116ꢀ4119.
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 5
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DOI: 10.1039/C5RA20197G
Synthesis and biological evaluation of novel hybrid compounds
between chalcone and piperazine as potential antitumor agents
Zewei Mao,a Xi Zheng,b Mengdi Zhang,a Yao Huang,b Chunping Wan*b and
Gaoxiong Rao*a
a School of Pharmacy, Yunnan University of Traditional Chinese Medicine, Kunming
650500, PR China
b Central laboratory, The NO.1 Affiliated Hospital of Yunnan University of
Traditional Chinese Medicine, Kunming 650021, PR China
O
O
N
N
N
X
Cell lines (IC50, μM)
A549 Hela SGC7901
X
7a
H
11.12 2.08 8.84
7.17 1.02 0.16
5.24 0.19 0.41
9.43 0.26 4.07
11.54 20.52 12.44
7b
F
7c
Cl
Br
7d
DDP
A series of novel hybrid compounds between chalcone and piperazine have been
synthesized, and their in vitro antitumor activity was evaluated against a panel of
human tumor cell lines by MTT assay. The results demonstrated that hybrid
compounds bearing acetophenone showed excellent antitumor activity, and 7c was
found to be the most potent compound.