263

Journal of Chemistry, Vol. 43 (2), P. 263 - 264, 2005

THREE MINOR ENT-KAUR-16-ENE-TYPE DITERPENE FROM
CROTON TONKINENSIS Gagnep.
Received 29
th
-Dec.-2004
Phan Minh Giang
1
, Hideaki Otsuka
2
, Phan Tong Son
1
1
Faculty of Chemistry, College of Natural Science, Vietnam National University, Hanoi
2
Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan
Summary
Three diterpenoids with ent-kaur-16-ene skeleton, namely, ent-11-acetoxy-7,14-
dihydroxykaur-16-en-15-one, ent-18-hydroxykaur-16-ene and ent-kaur-16-en-15-oxo-18-oic acid
were isolated from the MeOH extract of the dried leaves of Croton tonkinensis Gagnep.
(Euphorbiaceae) by successive chromatographic separation on silica gel, ODS gel and HPLC on
ODS. Their structures were established on the basis of spectroscopic analyses.
Keywords: Croton tonkinensis, Euphorbiaceae, ent-kaur-16-ene, diterpenoid.

The leaves of the endemic Vietnamese
medicinal plant Croton tonkinensis Gagnep.
(Euphorbiaceae) has been used in the treatment
of gastric and duodenal ulcers [1].

In our first
attempt to investigate the phytochemical
constituents of this plant, the main compound
was isolated as a new ent-kaur-16-en-15-one
[2]. Later, the acting mechanism of the herbal
remedy was discovered by us, at least in part,
through the ability of this compound to inhibit
the activation of the transcription factor nuclear
factor kappa B (NF-B) [3]. Guided by the
activity toward the inhibition of NF-B, further
new ent-kauranoids were isolated and
structurally determined [3, 4]. In addition, long-
chain alkyl alcohols [5] and flavonoid
glucosides [6] were isolated and structurally
identified. Our continuous search for the other
ent-kauranoids from this plant led to the
isolation of three minor diterpenoids 1 - 3, the
structure elucidation of which is described in
this paper. The extraction and isolation
procedures were carried out as described
previously [3, 4]. Preparative HPLC on ODS
columns using MeOH in water as solvent
systems was demonstrated to be an efficient
method in the purification of the minute amount
of compounds, which were concentrated in the
subfractions by successive column chroma-
tography on silica gel followed by column
chromatography on reversed-phase ODS gel.
Compounds 1 - 3 were obtained as amor-
phous powders in the quantities of ca. 2 mg of
each. In view of the paucity of material the
structures of these compounds were elucidated
mainly on the basis of the
1
H NMR spectra,
which were recorded on a high field NMR
instrument (JEOL JNM-ECP 500, 500 MHz).
Considering the
1
H NMR signals and the co-
occurrence of the other ent-kaurane-type
diterpenoids previously known from this plant
[2 - 4], 1 - 3 were also assumed to belong to the
ent-series of kaurane-type diterpenoids. The
1
H
NMR spectra of ent-kaur-16-enes 1, 2 and 3
were super-imposable with those reported in the
literature [7 - 9] of ent-11-acetoxy-7,14-
dihydroxy-kaur-16-en-15-one, ent-kaur-16-en-
15-oxo-18-oic acid, and ent-18-hydroxykaur-
16-ene, respectively.
264
AcO
O
OH
OH
H
H
HO
H
H
HOOC
O
H
H
3
1
2
Ent-11-acetoxy-7,14-dihydroxykaur-16-
en-15-one (1): Amorphous powder.
1
H NMR
(CDCl
3
, 500 MHz): 0.87 (3H, s, 19-CH
3
), 0.93
(3H, s, 18-CH
3
), 1.06 (3H, s, 20-CH
3
), 3.06 (1H,
br s, H-13), 4.39 (1H, br d, J = 12.0 Hz, H-7),
4.92 (1H, s, H-14), 5.38 (1H, s, H-17A), 6.11
(1H, s, H-17B).
Ent-kaur-16-en-15-oxo-18-oic acid (2):
Amorphous powder.
1
H NMR (CDCl
3
, 500
MHz): 1.13 (3H, s, 20-CH
3
), 1.18 (3H, s, 19-
CH
3
), 3.05 (1H, br s, H-13), 5.25 (1H, s, H-
17A), 5.94 (1H, s, H-17B).
Ent-18-hydroxykaur-16-ene (3): Amorphous
powder.
1
H NMR (CDCl
3
, 500 MHz): 0.76 (3H, s,
19-CH
3
), 1.03 (3H, s, 20-CH
3
), 2.39 (2H, d, J = 11.9
Hz, 2H-15), 2.61 (1H, br s, H-13), 3.10 (1H, d, J =
10.8, H-18A), 3.41 (1H, d, J = 10.8, H-18B), 4.92
(1H, s, H-17A), 4.94 (1H, s, H-17B).
The accumulation of ent-kaurane-type
diterpenoids in C. tonkinensis has been well
documented. The presence of three minor
diterpenoids, albeit in small amount, is an
additional proof for the significant role of ent-
kaurane-type diterpenoids as chemotaxonomic
markers of this plant. Ent-kaurane diterpenoids,
possessing an enone moiety at C-15/C-16 in
ring D, like compounds 1 and 2, were known as
good acceptors of nucleophiles, such as the
sulhydryl group of the cysteine residue, in a
Michael-type addition, and therefore they
display the inhibition of NF-B activity [3] and
apoptosis-inducing properties [10]. Moreover, a
recent study showed the inhibitory effect of
some ent-kaur-16-ene diterpenoids [11], thus
expanding our knowledge of the molecular
mechanism of NF-B inhibition of this class of
natural products. This research put an additional
scientific basis of the potential use of C.
tonkinensis in the treatment of inflammatory
diseases and cancer chemoprevention.
Acknowledgements: This research was
supported by the International Foundation for
Science, Stockholm, Sweden, through a Grant to
Dr. Phan Minh Giang, and the Basic Research
Program in Natural Science of Vietnam.
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