Secondary metabolites kurzichalcolactone A and B from ...

J. Siallagan et al.
ISBN 978-979-18962-0-7
Introduction
225
Proceeding of The International Seminar on Chemistry 2008 (pp. 225-228)
Jatinangor, 30-31 October 2008
Secondary metabolites kurzichalcolactone A and B from
Cryptocarya lucida Blume (Lauraceae)
Johnson Siallagan, Euis H. Hakim, Yana M. Syah, Lia D. Juliawaty,
Sjamsul A. Achmad, Lukman Makmur, Didin Mujahidin
Natural Products Research Group, Department of Chemistry, Institut Teknologi Bandung,
Jalan Ganeca 10 Bandung 40132, Indonesia
Abstract
Plants of Cryptocarya (Lauraceae) are known to contain 6-substituted 5,6-dihydro-2-pyrones,
flavanoids, alkaloids, and terpenoids. One of endemic plants of Kalimantan forest, C. lucida Blume,
has been selected for phytochemical study on its phenolic constituents. Fractionation of the methanol
extract of the stem bark of this plant, followed by purification of the phenolic fractions, afforded two
chalcone derivatives, named kurzichalcolactones A and B. The structures of both compounds were
determined by spectroscopic methods, including UV, IR, 1 H-NMR, 13 C-NMR data, as well as by
comparison with those reported data. Cytotoxic evaluation of both compounds against murine
leukemia P388 cells showed that their IC 50 to be 6.3 and 17.9 µg/mL, respectively.
Keywords: Kurzichalcolactone A and B, chalcone derivatives, Cryptocarya lucida Blume, cytotoxic
The genus Cryptocarya belongs to the pantropical
family Lauraceae and most of the species grow in the
Pacific-Asian tropical rainforests (Dumontet, 2001).
Many of these species have been examined for their
chemical constituents and revealed to contain
flavonoids, 6-alkyl- or 6-aryl-α-pyrones (Davies-
Coleman and Rivett, 1989; Collet et al., 1998),
lignans, terpenoids, and alkaloids (Juliawaty,
2000a,b). In continuation of our on going search for
cytotoxic compounds from Indonesia plants, we
investigated C. lucida Blume (Lauraceae), an endemic
plant collected at Bukit Bengkirai forest, East
Kalimantan Province, Indonesia. In this
communication, we report the isolation and structural
elucidation, as well as cytotoxicity against murine
leukemia P388 cells, of two chalcone derivatives,
kurzichalcolactones A (1) and B (2).
Materials and Methods
The following instruments were used: melting
points were determined by ‘micro melting point
apparatus’. Optical rotations, Perkin-Elmer 341
polarimeters in MeOH. UV spectra, Cary Varian 100
Conc and IR spectra, Perkin-Elmer Spectrum One FT-
IR spectrophotometers. 1 H and 13 C NMR spectra,
JEOL ECP500 spectrophotometers, 500 MHz ( 1 H)
and 125 MHz ( 13 C). The following adsorbents were
used for purification : Vacuum liquid chromatogaphy
(VLC), Si-gel 60 GF254 (Merck), Flash column
chromatography (FCC), Si-gel G60 (230-400 mesh)
(Merck), Radial chromatography (RC), Si-gel 60
PF254 (Merck), and TLC plates, Kieselgel 60 F254 0.25
mm (Merck) thick were used. All solvents distilled
technical qualities.
Plant material. Samples of the stem bark of C.
lucida was collected in Oktober 2005 from the Bukit
Bengkirai forest, Balikpapan, East Kalimantan,
Borneo island, Indonesia. A herbarium specimen has
been deposited at the Herbarium Bogorienses, Center
of Biological Research and Development, National
Institute of Science, Bogor, Indonesia.
Extraction and isolation. The milled stem bark of
C. lucida (3.1 kg) was extracted successively with
with methanol (3 x 24h x 6L) at room temperature.
The residue give the crude extract (120 g) which was
then partitioned by use of n-hexane to produce extract
(3.7 g) and than EtOAc extract (61.5 g). Next, the
EtOAc extract was subject to chromatography on
silica gel and eluted successively with n-hexane, nhexane/acetone,
acetone, and MeOH in the order of
increasing polarity. Extract EtOAc was subjected to
VLC with column ∅ 10 cm (3 x 20g), adsorbent Sigel
(200 g) and eluted with mixture hexane-acetone
(9:1; 8:2; 7.5:2.5; 6:4; 1:1), acetone 100 %, and
MeOH (100 %), to give 7 fraction (A-G) were 1.6 g,
4.1 g, 5.1 g, 5.4 g, 16.2 g, 11.4 g, and 5.8 g,
respectively. The major component of fraction E.
Compound 1 (40 mg) and 2 (35 mg) were obtained by
purification of E (293 mg) using radial
chromatography (Si-gel, hexane:acetone = 6.5:3.5 and
hexane:acetone = 6:4). These compounds were all
determined by spectral methods and compare with
corresponding data reported in the literature.
Kurzichalcolactone A (1), yellow powder : [α] 20 D -
47 0 (c= 0.1 Me-OH) UV absorption λmaks (nm)
MeOH) (log ε) 206 (2.34), 239 (1.20), 355 (1.12) nm,
showed bathochromic shift (MeOH + NaOH).

J. Siallagan et al.
number Compound 1(lit.)
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
1’
2’
3’
4’
5’
6’
OH-5
δC δH
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Proceeding of The International Seminar on Chemistry 2008 (pp. 225-228)
Jatinangor, 30-31 October 2008
Table 1 NMR Data for Kurzichalcolactones A (1) and B (2) in (acetone-d6)
143.3 7.72 d (16)
128.9 8.22 d (16)
193.4
162.6
97.0 6.11 s
167.4
105.5
159.6
106.2
175.1
34.6 2.22 tl (7)
22.1 1.50-1.80 m
36.1 1.50-1.80 m
71.4 3.78 m
36.5 1.50-1.80 m
24.5 3.58 dd (3; 2.5)
34.4 2.12 dd (13; 2.5)
2.20 dd (13; 3)
100
132.4 7.12 d (16)
131.0 6.68 d (16)
137.5
128.3 7.53 m
129.5 7.36 m
129.5 7.36 m
129.5 7.36 m
128.3 7.53 m
136.7
130.0 7.62 m
130.0 7.46 m
130.0 7.46 m
130.0 7.46 m
130.0 7.62 m
14.10 s
Compound 2(lit.)
δC δH
143.1 7.72 d (15.6)
128.6 8.31 d (15.6)
193.8
163.5
94.9 6.09 s
164.5
106.2
161.0
106.1
174.7
34.7 2.30 tl (7.0)
23.9 1.50-1.80 m
35.9 1.50-1.80 m
71.0 3.70 m
36.1 1.50-1.80 m
21.9 3.55 m
34.2 2.17 dd (12.9; 2.7)
1.91 dd (12.9; 3.1)
99.4
131.4 6.94 d (16.1)
131.2 6.42 d (16.1)
137.3
127.8 7.54 dl
129.3 7.37 tl (7.5)
129.0 7.29 tl (7.5)
129.3 7.37 tl (7.5)
127.8 7.54 m
136.6
129.6 7.71 m
130.0 7.46 m
130.9 7.46 m
130.0 7.46 m
129.6 7.71 m
14.50 s
Compound 1 (isolated)
δC δH
142.6 7.71 d (15.9)
128.6 8.19 d (15.9)
193.0
159.5
96.5 6.09 s
166.7
105.0
162.0
105.7
173.1
34.1 2.04 m
21.7 1.50-1.60 m
35.6 1.60-1.70 m
70.9 3.78 m
36.1 1.70-1.80 m
24.1 3.57 m
33.9 2.17 dd (13; 2.5)
2.29 dd (13; 2.5)
99.6
131.9 7.09 d (16)
130.6 6.66 d (16)
137.0
127.8 7.51 m
129.0 7.34 m
129.0 7.34 m
129.0 7.34 m
127.8 7.51 m
136.2
130.5 7.54 m
129.6 7.38 m
129.6 7.38 m
129.6 7.38 m
130.0 7.54 m
14.13 s
Compound 2
(isolated) δH
7.81 d (15.9)
8.29 d (15.9)
6.07 s
2.22 tl (7)
1.52-1.62 m
1.69-1.76 m
3.70 m
1.89-1.91 m
3.54 dd (3; 2.5)
2.16 dd (12.8; 2.29)
2.29 dd (12.8; 2.29)
6.92 d (15.9)
6.41 d (15.9)
7.52 m
7.35 m
7.35 m
7.35 m
7.52 m
7.70 m
7.43 m
7.43 m
7.43 m
7.70 m
14.50 s
Table 2 IC 50 values of some chalcolactone against murine leukemia P-388 cells
Compounds Classes of compound IC50 (µg/mL)
Kurzichalcolactone A (1)
Kurzichalcolactone B (2)
Artonin E
Chalcolactone
Chalcolactone
3-prenylflavone
a For significant activity, an IC50 < 4.0 µg/mL is required.
b Positive control
IR bands spectrum (KBr) V maks (cm -1 ), 3171 cm -1 (br,
OH), 2938 cm -1 (CH-aliphatic), 1707, 1626, cm -1
(lactone, C=O conjugated systems), 1344, 1227, 1158,
1082, 1032, 975, 940, 749, and 692 cm -1 . 1 H NMR
(500 MHz, acetone-d6): 8.19 (1H, d, J= 15.9), 7.71
(1H, d, J= 15.9), 7.54 (2H, m), 7.51 (2H, m), 7.38
(3H, m), 7.34 (3H, m), 7.09 (1H, d, J= 16), 6.66 (1H,
d, J= 16), 6.09 (1H, s), 3.78 (1H, m), 3.70 (1H, m),
2.29 (1H, dd, J= 13; J= 3), 2.17 (1H, dd, J= 13; J=
6.3
17.9
0.6
2.5), 2.04 (1H, m), 1.70-1.80 (1H, m), 1.60-1.70 (1H,
m), 1.50-1.60 (1H, m), 14.13 (1 OH). 13 C NMR see
Table 1.
Kurzichalcolactone B (2) yellow powder : [α] 20 D -
88 0 (c= 0.1 Me-OH) UV absorption λmaks (nm)
MeOH) (log ε) 205 (1.28), 236 (0.61), 254 (0.55),
344 (0.74) nm, showed bathochromic shift (MeOH +
NaOH). IR bands spectrum (KBr) V maks (cm -1 ) 3224
cm -1 (br, OH), 2937 cm -1 (CH-aliphatic), 1706, 1626,

J. Siallagan et al.
cm -1 (lactone, C=O conjugated systems), 1338, 1227,
1139, 1084, 970, 933, 749, and 691 cm -1 . 1 H NMR
(500 MHz, acetone-d6): 8.29 (1H, d, J= 15.9), 7.81
(1H, d, J= 15.9), 7.70 (2H, m), 7.52 (2H, m), 7.43
(3H, m), 7.35 (3H, m), 6.92 (1H, d, J= 15.9), 6.41
(1H, d, J= 15.9), 6.07 (1H, s), 3.70 (1H, m), 3.54 (1H,
dd, J= 3; 2.5), 2.29 (1H, dd J= 12.8; 2.29), 2.22 tl (7),
2.16 (1H, dd, J= 12.8; J= 2.9), 2.04 (1H, m), 1.89-
1.91 (1H, m), 1.69-1.76 (1H, m), 1.60-1.70 (1H, m),
1.52-1.62 (1H, m), 14.50 (1 OH).
Results and Discussion
The dried milled stem bark of C. lucida was
macerated with methanol, and the methanol extract
was partitioned into n-hexane and EtOAc,
respectively. The EtOAc fraction was refractionated
by VLC on silica gel, and the fraction contain of
phenolic compounds was purified by radial
chromatography on silica gel to give
kurzichalcolactones A (1) and B (2).
Compound 1 showed IR absorption maxima at
3171, 1707, and 1626 cm -1 , indicated the presence of
hydroxyl, lactone, and conjugated ketone
functionalities. A comparison of the NMR spectra of 1
with those of kurzichalcolactone B (2), isolated from
this plant and previously from C. kurzii and C.
obovata revealed the strong structural similarity of
these two compounds (Table 1) and indicated that
compound 1 was a stereoisomer of 2. Analysis of the
1 H- 1 H COSY, HMQC, and HMBC spectra permitted
the assignment of all proton and carbon signals for 1.
There are three asymmetric carbons in compounds 1
and 2, at the 15, 17, and 19 positions. The signals for
H-15 and H-17 in the 1 H NMR spectrum of both 1 and
2 are essentially identical, suggesting that the two
isomers possess the same configuration at these
positions and only differ by the α or β position at the
C-19 position. Moreover, the small coupling constants
observed for H-17 in compounds 1 and 2 indicated an
equatorial position for H-17 in both compounds,
whereas the H-15 peak appearance.
Kurzichalcolactones A (1) and B (2) had been
previously isolated from leaves C. kurzii (Fu, 1993)
and C. obovata (Dumontet, 2004), respectively.
Biogenetically, 1 could be formed by a coupling of
5,7,9-trihydroxychalcone with a tetraketide derivative
of cinnamoyl to form kurzilactone, which is also
isolated from C. Kurzii. Phenol coupling of this
lactone would lead to a linear intermediate, which
would cyclize twice to form the C8-C17 bond as well
as the pyran ring.
227
Proceeding of The International Seminar on Chemistry 2008 (pp. 225-228)
Jatinangor, 30-31 October 2008
O
O
O OH 20
OH
19
18
OH
O
kurzichalcolactone A (1) : 19α- or 19β-OH
kurzichalcolactone B (2): 19β- or 19α-OH
Interesting biological activities have been
described for many individual chalcone metabolites
isolated from Cryptocarya. In a preliminary bioassays,
the fruit and trunk bark extracts of C. obovata showed
56% and 23% inhibition at 10 µg/mL the ethanolic
extracts against human nasopharyngeal KB cells
(Dumontet, 2004).
In further study, kurzichalcolactone A (1)
exhibited weak cytotoxicity against the same cells
with IC50 value of 15 µg/ml (Fu, 1993). In our study,
we evaluated cytotoxic properties of compounds 1 and
2 against murine leukemia P-388 cells (Table. 2)
using MTT-assay method, with artonin E was used as
positive control. As shown in Table 2, compound 1
was more cytotoxic than compound 2. Thus, different
stereochemical arrangement could have a different
cytotoxic properties.
Conclusions
Two chalcolactones, i.e. kurzichalcolactones A (1)
and B (2) have been isolated from the methanol
extract of the stem bark of C. lucida. The structures of
these compounds were determined based on spectral
analysis and by comparison with those reported data.
The cytotoxic activities of both compounds were
evaluated against murine leukaemia P388 cells and
showed their IC 50 values were 6.3 and 17.9 µg/ml,
respectively.
Acknowledgements
We would like to thank the Dinas P dan P
PEMDA Papua and Rector of Cenderawasih
University Jayapura for the financial support, and Dr.
Jalifah Latip from UKM for measurements for
spectral data. We also very thankful for the Herbarium
Bogoriense staffs, Bogor, for the identification of the
sample.

J. Siallagan et al.
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