205

A novel limonoid from the seeds of Chisocheton

siamensis
Surat Laphookhieo, Wisanu Maneerat, Sorwaporn Koysomboon,
Rattana Kiattansakul, Kan Chantrapromma, and John Keith Syers

Abstract: Analysis of an acetone/hexane (1:1) extract of the seeds of Chisocheton siamensis led to the isolation of a
novel limonoid, chisosiamensin (1), along with five known limonoids, dysobinin (2), azadiradione (3), mohinin (4),
epoxyazadiradione (5), and 6 -acetoxyepoxyazadiradione (6). Their structures were characterized by spectroscopic
methods, including UV, IR, NMR, and MS. All isolated limonoids were reported for the first time as secondary metabolites of C. siamensis.
Key words: Chisocheton siamensis, Meliaceae, chisosiamensin, limonoids.
Rsum : Lanalyse dun extrait par un mlange actone-hexane (1:1) de graines de Chisocheton siamensis a permis
dextraire un nouveau limonode, la chisosiamensine (1), aux cts de cinq limonodes connus, la dysobinine (2),
lazadiradione (3), la mohinine (4), lpoxyazadiradione (5) et la 6 -actoxypoxyazadiradione (6). On a caractris
leurs structures par des mthodes spectroscopiques, incluant lUV, lIR, la RMN et la SM. Tous les limonodes isols
ont t identifis pour la premire fois comme mtabolites secondaires du C. siamensis.
Mots-cls : Chisocheton siamensis, Meliaceae, chisosiamensine, limonodes.
[Traduit par la Rdaction]

Laphookhieo et al.

208

Introduction

Meliaceae plants are known to be rich sources of

limonoids. A number of limonoids have been isolated from
several genera of Meliaceae and some of these exhibit
antimalarial (1, 2), cytotoxic (3, 4), antiprotozoal (5), and
antifeedant (6, 7) activities. Chisocheton siamensis is one of
the Meliaceae plants, which is found in the northern part of
Thailand. This plant has been used by local Thai people as
anti plant pathogen and antimalarial. In the continuation of
our research on secondary metabolites from Thai medicinal
plants (8, 9), we have studied an acetone/hexane (1:1) extract of the seeds of C. siamensis identified a new limonoid,
chisosiamensin (1), along with five known compounds (2
6). In this paper, we describe the isolation and identification
of a new and known limonoids isolated from the seeds of C.
siamensis. The plausible biogenetic pathway of the new
compound 1 and limonoids 46 is also reported.

H
O

H
O

Azadiradione (3)

Dysobinin (2)

Chisosiamensin (1)

O
O

H
O
O

H
H

O
O

O
O

O
O

O
O

Epoxyazadiradione (5)

Mohinin (4)

6 -Acetoxyepoxyazadiradione (6)

Received 25 September 2007. Accepted 16 December 2007.

Published on the NRC Research Press Web site at
canjchem.nrc.ca on 1 February 2008.
S. Laphookhieo,1 W. Maneerat, and J.K. Syers. School of
Science, Mae Fah Luang University; Tasud, Muang, Chiang
Rai 57100, Thailand.
S. Koysomboon and R. Kiattansakul. Faculty of Arts and
Sciences, Prince of Songkla University, Muang, Suratthani
84100, Thailand.
K. Chantrapromma. School of Science, Walailak University,
Thaiburi, Thasala, Nakhon Si Thammarat 80160, Thailand.
1

Corresponding author (e-mail: surat@mfu.ac.th or

laphookhieo@yahoo.com).

Can. J. Chem. 86: 205208 (2008)

H
O

O
OH

Nimonol (7)

Results and discussion

Soaking the seeds of C. siamensis in acetone/hexane (1:1)
for 3 days provided a crude extract. This crude extract was
subjected to a succession of chromatographic procedures, in-

doi:10.1139/V07-155

2008 NRC Canada

206
Table 1. 1H,

Can. J. Chem. Vol. 86, 2008

13

C, and DEPT spectral data of chisosiamensin (1) and nimonol (7).

Chisosiamensin

Nimonol

C/H

H (J in Hz)

H (J in Hz)

DEPTa

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
28
29
30
7-OCOCH3
7-OCOCH3

158.1
125.4
204.6
39.9
158.2
129.7
74.5
44.1
38.6
42.8
16.5
33.0
47.1
158.8
119.6
34.3
51.6
19.0
27.0
124.5
139.6
111.0
142.5
21.1
27.3
20.6
170.1
21.2

7.15
5.84

5.33
5.27

2.28

1.75
1.68

5.37
2.35
2.80
0.78
1.07

7.23
6.27
7.37
1.19
1.22
1.07

1.95

157.40
126.14
205.96
40.51
49.88
68.08
79.08
45.43
37.13
43.11
16.39
32.72
47.08
158.53
119.55
34.30
51.12
20.84
27.08
124.37
139.64
110.94
142.57
20.23
31.87
20.79
172.32
21.16

7.12 (d, J = 10.06)

5.90 (d, J = 10.06)

2.21 (d, J = 11.65)

4.38 (dd, J = 11.65, 2.37)
5.36 (d, J = 2.37)

2.21 (m)

1.82 (m)
1.82 (m)

5.42 (dd, J = 1.82, 2.84)

1.7 (m), 2.4 (m)
2.83 (m)
0.82 (s)
1.14 (s)

7.26 (m)
6.29 (m)
7.39 (m)
1.27 (s)
1.31 (s)
1.41 (s)

2.05 (s)

CH
CH
C
C
C
CH
CH
C
CH
C
CH2
CH2
C
C
CH
CH2
CH
CH3
CH3
C
CH
CH
CH
CH3
CH3
CH3
C
CH3

(d, J = 10.2)
(d, J = 10.2)

(m)
(br m)
(m)
(m), 2.03 (m)
(m), 1.95 (m)

(br m)
(m)
(dd, J = 7.8, 10.5)
(s)
(s)
(br s)
(br m)
(br m)
(s)
(s)
(s)
(s)

The chisosiamensin data were analyzed by DEPT 90 and 135.

cluding quick column chromatography (QCC) and column

chromatography (CC) and crystallization to afford a new
limonoid, chisosiamesin (1), along with five known
limonoids (26).
The 1H NMR spectra of limonoids isolated from the seeds
of C. siamensis that were tetranortriterpenoids indicated that
all compounds contained an ,-unsaturated ketone at ring
A and a -furan moiety at C-17 of ring D. Three of these
were connected with two acetoxyl groups at C-6 and C-7 in
ring B, which existed in compounds 2, 4, and 6, whereas
limonoids 1, 3, and 5 consisted of an acetoxyl group at C-7.
In addition, compounds 3 and 4 also contained an
additional ,-unsaturated ketone at ring D, while compounds 5 and 6 had an epoxy ketone instead of an ,unsaturated ketone.
Chisosiamensin (1) was isolated as a colorless viscous oil
with a molecular ion peak at m/z 435.4526 [M + H]+ in
HREIMS, corresponding to the molecular formula
C28H34O4. The UV spectrum indicated the presence of ,unsaturated ketone at 218 nm, while the IR spectrum of this
compound showed two carbonyl functionalities at 1750 and
1680 cm1. Compound 1 was classified as tetranortriterpenoidal limonoid by the presence of five methyl singlets at
: 0.78 (Me-18), 1.07 (2 Me, Me-19, and Me-30), 1.19

(Me-28), and 1.22 (Me-29) and a -furan moiety at : 6.27

(s, H-22), 7.23 (s, H-21), and 7.37 (s, H-23) in the 1H NMR
spectrum (10, 11). The 1H NMR and 13C NMR spectral data
of 1 (Table 1) were similar to nimonol (7), isolated from
Azadirachta indica (10), except that 1 showed an olefinic
proton at : 5.33 (m), which was connected to sp2 carbon at
: 129.7 (C-6) in the HMQC experiment, instead of the two
methine protons (H-5 and H-6) of nimonol (Table 1). These
results indicated that compound 1 consisted of a double at
C-5/6. Therefore, chisosiamensin was identified as 1, a dehydrated derivative of nimonol.
The remaining limonoids (26) were characterized as
dysobinin (2) (12), azadiradione (3) (2), mohinin (4) (13),
epoxyazadiradione (5) (14), and 6 -acetoxyepoxyazadiradione (6) (15) by the analysis of 1D and 2D NMR
data and by comparison with their reported physical and
spectroscopic data. It should be noted that the plausible
biogenetic pathway of three limonoids (1, 4, and 6) could be
derived from dysobinin (2) (Scheme 1). The elimination of
AcOH of dysobinin produced chisosiamensin, whereas the
hydroxylation at C-16 of dysobinin followed by alcohol
oxidation yielded mohinin (4). Epoxidation at C-14/15 of
mohinin (4) gave 6-acetoxyepoxyazadiradione (6). In the
case of epoxyazadiradione (5), this compound could be de 2008 NRC Canada

Laphookhieo et al.

207

Scheme 1. Plausible biogenetic pathway of compounds 1 and 46.

rived from the epoxidation at C-14/15 of azadiradione (3)

(Scheme 1). Finally, the control experiment of compound 2
(dysobinin) was done. Unfortunately, no compound 1
(chisosiamensin) was observed from this experiment. Therefore, compound 1 was a natural product produced from the
seeds of this plant.
It is worth noting that a number of limonoids have been
isolated from the Meliaceae species. However, less than 30
compounds have been isolated from the genus Chisocheton.
In this study, we isolated an additional novel limonoid from
the seeds of C. siamensis and all isolated limonoids were reported for the first time as secondary metabolites of C.
siamensis.

Experimental section
General
Melting points were determined using a FisherJohn melting point apparatus. The optical rotation []D values were
determined with a JASCO P-1020 digital polarimeter. UV
spectra were measured with a UV-160A spectrophotometer
(Shimadzu). The IR spectra were measured with a
PerkinElmer FTS FT-IR spectrophotometer. The 1H NMR
and 13C NMR spectra were recorded using a 300 MHz
Bruker FTNMR Ultra Shield spectrometer. Chemical shifts
were recorded in parts per million () in CDCl3 with
tetramethylsilane (TMS) as the internal reference. The EIMS
was obtained from a MAT 95 XL mass spectrometer. Quick
column chromatography (QCC) and column chromatography
(CC) were carried out on silica gel 60 H (Merck, 540 m)
and silica gel 100 (Merck, 63200 m), respectively.
Precoated plates of silica gel 60 F254 were used for the analytical procedure.
Plant material
Seeds of C. siamensis were collected from Queen Sirikit
Garden, Mae Rim District, Chiang Mai Province, northern
Thailand, in February, 2006. The botanical identification
was made by Dr. Prachaya Srisanga, botanist at Queen
Sirikit Garden. Voucher specimens are deposited in the

Herbarium of Queen Sirikit Garden, Mae Rim District,

Chiang Mai, Thailand.
Extraction and isolation
Seeds of C. siamensis (600 g) were extracted with
acetone/hexane (1:1) over the period of 3 days at room temperature. The mixture was filtered and concentrated under
reduced pressure to provide the crude extract (62.21 g). This
crude extract was subjected to quick column chromatography (QCC, column size: 12 cm 16 cm) over silica gel and
eluted with a gradient of EtOAc/hexane to afford 13 factions
(A1A13). Fraction A2 (435.9 mg) was subjected to column
chromatography (CC) using 15% EtOAc/hexane to give
compound 1 (51.2 mg). Fraction A4 (4.0 g) upon standing at
room temperature yielded compound 2 (3.30 g). Compound
5 (227.0 mg) was obtained from fraction A6 (576.9 mg) after repeated CC with 25% EtOAc/hexane. Purification of
fraction A8 (1.01 g) by CC with 25% EtOAc/hexane gave
compounds 6 (17.2 mg) and 3 (31.2 mg). Compound 4
(18.2 mg) was obtained from fraction A10 (65.5 mg) by repeated CC using 5% EtOAc/CH2Cl2 as eluent.
Control experiment of dysobinin (2)
A solution of dysobinin (10 mg) and 1:1 acetone/hexane
(2 mL) was stirred at room temperature for three days. The
solution mixture was monitored by TLC (15% EtOAc/hexane)
and only dysobinin was observed on the TLC.
Chisosiamensin
Colorless viscous oil. []27D +49.0 (c 0.04, MeOH). UV
max (log ) (MeOH): 218 (4.37). IR (neat): 1750, 1680. For
1
H NMR and 13C NMR see Table 1. EI-MS m/z: 435 (87)
[M + H]+. HR-EI-MS m/z: 435.4526 [M + H]+.

Acknowledgments
Surat Laphookhieo thanks the Bioresources Research
Network, National Center for Genetic and Engineering and
Biotechnology (grant number BRN 003 G-49), and Mae Fah
Luang University for financial support.
2008 NRC Canada

208

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