Bioactive Constituents from Michelia champaca

Abstract

(-)-Anonaine (1), (-)-asimilobine (2), (-)-nuciferine (3), (-)-anolobine (4), (-)-romerine (5), (-)-N-acetylanonaine (6), liriodenine (7), (+)-syringaresinol (8), N-trans-feruloyltyramine (9), N-cis-feruloyltyramine (10), scopoletin (11), 4-acetonyl-3,5-dimethoxy-p-quinol (12), vanillin (13), vanillic acid (14), syringic acid (15), beta-sitosterol (16) and stigmasterol (17) were isolated from branches of Michelia champaca L. In addition, a cell proliferation assay of five of the isolated compounds on human breast and lung cancer cells showed that liriodenine (7) was the strongest inhibitor.

Full text

Bioactive Constituents from Michelia champaca
Yu-Ting Yeha, Jin-Cherng Huangb, Po-Lin Kuoc,* and Chung-Yi Chena,*
aSchool of Medical and Health Sciences, Fooyin University, Kaohsiung 831, Taiwan, R.O.C.
bDepartment of Forest Products Science and Furniture Engineering, National Chiayi University,
Chiayi 600, Taiwan, R.O.C.
cInstitute of Clinical Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan, R.O.C.
xx377@mail.fy.edu.tw; kuopolin@seed.net.tw
Received: June 1st, 2011; Accepted: June 11th, 2011
(-)-Anonaine (1), (-)-asimilobine (2), (-)-nuciferine (3), (-)-anolobine (4), (-)-romerine (5), (-)-N-acetylanonaine (6), liriodenine (7),
(+)-syringaresinol (8), N-trans-feruloyltyramine (9), N-cis-feruloyltyramine (10), scopoletin (11), 4-acetonyl-3,5-dimethoxy-p-quinol (12),
vanillin (13), vanillic acid (14), syringic acid (15), -sitosterol (16) and stigmasterol (17) were isolated from branches of Michelia champaca
L. In addition, a cell proliferation assay of five of the isolated compounds on human breast and lung cancer cells showed that liriodenine (7)
was the strongest inhibitor.
Keywords: Michelia champaca L., aporphine, liriodenine.
Breast cancer is the most common female cancer,
comprising 23% of all cancers, with more than 1 million
new cases per year. Effective chemopreventive treatments
for breast cancer are still needed to increase available
regimens and advance the efficacy of chemotherapy
[1a-1c]. Lung cancer is one of the most common malignant
and devastating of human tumors. Non-small cell lung
cancer (NSCLC) A549 cells commonly develop resistance
to radiation and chemotherapy [1d,1e]. Nevertheless, a
great number of tumor histotypes show a low or very low
response to such treatment, and only some display optimal
results after its application. This is the main reason that the
search for new anticancer compounds that are more
effective against unresponsive tumors, with reduced
adverse effects, is considered an important step in
improving cancer chemotherapy [1f,1g].
Michelia champaca L. is a large evergreen tree, native to
South and Southeast Asia, which is primarily cultivated for
its timber; its aril-covered seeds are highly attractive to
birds [2a]. Previous phytochemical studies led to the
isolation of ushinsunine, oxoushinsunine, magnoflorine,
parthenolide, costunolide, dihydroparthenolide, micheli-
nolide, 11,13-dehydrolanuginolide, magnograndiolide,
michampanolide, quercetin, benzaldehyde 4-O-β-D-gluco-
pyranoside, syringaresinol, and N-acetylanonaine [2b-2f].
To continue the search for novel agents from Magnoliaceous
plants, the branches of M. champaca were chosen for the
first time for phytochemical investigation. In this paper,
we report the isolation of seventeen compounds, including
six aporphines: (-)-anonaine (1) [2g], (-)-asimilobine (2)
[2h], (-)-nuciferine (3) [2i], (-)-anolobine (4) [2j], (-)-
romerine (5) [2h], and (-)-N-acetylanonaine (6) [2h]; one
N
O
O
R
H
5: R = CH3
6: R = COCH3
N
O
O
O
7
N
O
H
HO
H3CO
OH
9
0
20
40
60
80
100
120
Control
Liriodenine
Vanillin
(-)-Roemerine
(-)-N-Acety lanonaine
N-trans-Feruloyltyrami ne
Cell proliferation (% of control)
A
549
*
***
0
20
40
60
80
100
120
Control
Liriodenine
Vanillin
(-)-Roemerine
(-)-N-Acetylanonaine
N-trans-Ferulo yltyramine
Cell proliferation ( % of control)
MDA-MB-231
**
*
*
Figure 1: Antiproliferative effect of liriodenine, vanillin, (-)-roemerine, (-)-N-
acetylanonaine, and N-trans-feruloyltyramine on A549 cells and MDA-MB-231 cells.
oxoaporphine: liriodenine (7) [2k]; one lignan: (+)-
syringaresinol (8) [2e]; one amide: N-trans-
feruloyltyramine (9) [2l], N-cis-feruloyltyramine (10) [2l];
one coumarin: scopoletin (11) [2m]; one quinol: 4-
acetonyl-3,5-dimethoxy-p-quinol (12) [2n]; three
benzenoids: vanillin (13) [2o], vanillic acid (14) [2g], and
syringic acid (15) [2p]; and two steroids: -sitosterol (16)
[2q] and stigmasterol (18) [2h]. All compounds, except 6
and 8, are found for the first time from this plant [2b-2f].
To evaluate the relative anticancer activity of liriodenine,
vanillin, (-)-roemerine, (-)-N-acetylanonaine, and N-trans-
feruloyltyramine, we investigated their proliferation
inhibition effects in A549 human lung adenocarcinoma
cells and MDA-MB-231 human breast adenocarcinoma
cells. As shown in Figure 1, the maximal effect on
proliferation inhibition was observed with 20 µM
liriodenine at 48 h, which inhibited proliferation of
A549 and MDA-MB-231 cells by 54.4% and 51.7%,
NPC Natural Product Communications 2011
Vol. 6
No. 9
1251 - 1252

1252 Natural Product Communications Vol. 6 (9) 2011 Cheng et al.
respectively. In contrast, 20 µM (-)-N-acetylanonaine at 48
h did not affect cell viability of A549 and MDA-MB-231
cells.
Experimental
Cell culture and cell proliferation assay: A549 human
lung adenocarcinoma cells and MDA-MB-231 human
breast adenocarcinoma cells were cultured in Minimum
essential medium supplemented with 10% FBS, 100 units/
mL of penicillin G, 100 µg/mL of streptomycin, and 0.25
µg/mL of amphotericin B (GIBCO BRL, Gaithersburg,
MD, USA). All test compounds were dissolved in DMSO
and stored at -20°C. Control cultures received the carrier
solvent (0.1% DMSO). Cell proliferation was assessed by
Premixed WST-1 Cell Proliferation Reagent (Clontech
Laboratories Inc., Mountain View, CA, USA) according to
the manufacturer’s instructions.
Purification of active ingredients from M. champaca:
Michelia champaca L. was collected from Pingtung
County, Taiwan in December, 2009. A voucher specimen
was characterized by Dr Jin-Cherng Huang of the National
Chiayi University, and deposited in Fooyin University,
Kaohsiung County, Taiwan.
The air-dried branches of M. champaca (11.0 kg) were
extracted with MeOH (80 L x 6) at room temperature and
the MeOH extract (181.5 g) obtained after concentration
under reduced pressure. This was chromatographed over
silica gel (800 g, 70-230 mesh) using n-hexane/acetone as
eluent to produce 5 fractions. These (supporting material)
were subjected to repeated Si gel CC and preparative
TLC to afford β-sitosterol (10.41 mg) and stigmasterol
(6.67 mg) from fraction 1, liriodenine (12.53 mg), (-)-N-
acetyl- anonaine (6.01 mg), scopoletin (4.42 mg), (+)-
syringaresinol (5.62 mg) and vanillin (4.35 mg) from
fraction 2, vanillic acid (4.26 mg) from fraction 3,4-
acetonyl-3,5-dimethoxy-p-quinol (12.54 mg), syringic acid
(15.03 mg), N-trans-feruloyltyramine (5.36 mg), N-cis-
feruloyltyramine (2.25 mg), and (-)-romerine (8.47 mg)
from fraction 4, and (-)-anonaine (9.02 mg), (-)-
asimilobine (11.08 mg), (-)-nuciferine (6.71 mg) and (-)-
anolobine (5.37 mg) from fraction 5.
Supporting data: Detailed isolation procedure.
Acknowledgments - This investigation was supported by
a grant from the National Science Council of the Republic
of China (NSC 97-2320-B-242-002-MY3).
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