This article was downloaded by: [University of Hong Kong Libraries]

On: 15 November 2014, At: 04:27

Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered
office: Mortimer House, 37-41 Mortimer Street, London Wl T 3JH, UK

Natural Product Letters

Publication details, including instructions for authors and
subscription information:
http://www.tandfonline.com/loi/gnpl 19

Two Minor, Cytotoxic Caged

Xanthonoids from Garcinia
gaudichaudii
Xiao-Hua Wu ab, B. K. H. Tan a, Shu-Geng Cao b, K.-Y. Sim b &
S. H. Goh b
a Departments of Pharmacology , 119260, Singapore
b Chemistry , National University of Singapore , 119260,
Singapore
Published online: 04 Oct 2006.

To cite this article: Xiao-Hua Wu, B. K. H. Tan, Shu-Geng Cao, K.-Y. Sim & S. H. Goh (2000) Two
Minor, Cytotoxic Caged Xanthonoids from Garcinia gaudichaudii , Natural Product Letters, 14:6,
453-458, DOI: 10.1080/10575630008043784
To link to this article: http:/ /dx.doi.org/10.1080/10575630008043784

PLEASE SCROLL DOWN FOR ARTICLE

Taylor & Francis makes every effort to ensure the accuracy of all the information (the
"Content") contained in the publications on our platform. However, Taylor & Francis,
our agents, and our licensors make no representations or warranties whatsoever as to
the accuracy, completeness, or suitability for any purpose of the Content. Any opinions
and views expressed in this publication are the opinions and views of the authors,
and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content
should not be relied upon and should be independently verified with primary sources
of information. Taylor and Francis shall not be liable for any losses, actions, claims,
proceedings, demands, costs, expenses, damages, and other liabilities whatsoever
or howsoever caused arising directly or indirectly in connection with, in relation to or
arising out of the use of the Content.

This article may be used for research, teaching, and private study purposes. Any
substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing,
systematic supply, or distribution in any form to anyone is expressly forbidden. Terms
& Conditions of access and use can be found at http://www.tandfonline.com/page/
terms-and-conditions
Natural Product Letters
Volume 14(6), pp. 453-458
Reprints available directly from the Publisher
Photocopying permitted by license only
© 2000 OPA (Overseas Publishers Association) N.V.
Published by license under
the Harwood Academic Publishers imprint,
part of The Gordon and Breach Publishing Group.
Printed in Malaysia.

TWO MINOR, CYTOTOXIC CAGED XANTHONOIDS FROM

GARCINIA GAUDICHAUDII

XIAO-HUA wu,« B. K. H. TAN,tSHU-GENG CAO,§

K.-Y. SIM1 AND s. H. Gow·

Departments of Pharmacology' and Chemistry,§

National University of Singapore, Singapore 119260

(Received 23rd February 2000)

Abstract: Bioassay-guided fractionation allowed the isolation of two minor, caged

cytotoxic tetraprenylated xanthonoids, gaudichaudiones I and J (1 and 2), from the
leaf extract of Garcinia gaudichaudii. The structures were determined by spectral
analysis which provided good agreement with the previouslyisolated gaudichaudi-
one B(3).

Key Words: Garcinia, prenylated xanthonoids, cytotoxic, gaudichaudiones.

In continuation of our study on cytotoxic natural products from the Malaysian

Garcinia species, which was found to be a rich source of caged tetraprenylated
xanthonoids, 1·6 we have further studied the leaf extracts of Garcinia gaudichaudii
(Guttiferae). Two new caged cytotoxic tetraprenylated xanthonoids, gaudichaudi-
ones I and J (1 and 2), were isolated. In this paper we describe the isolation of 1 and
2, and present their structural elucidation using UV, IR, mass spectrometry, and
NMR spectroscopy and also determined their cytotoxic activity.
Our detailed study on the leaf extracts of G. gaudichaudii indicated that a
cytotoxic fraction contained two minor compounds. This fraction was separated by
repeated chromatography and found to contain two new tetraprenylated xan-
thonoids, gaudichaudiones I and J (1 and 2). These two compounds, being
diastereomers, have similar properties and were separable only by HPLC. Their
NMR spectra indicated that they were related diastereoisomers of gaudichaudiones
A-H, isolated earlier.5·6 Being minor compounds, the isolated materials were just
sufficient to provide basic NMR spectral data and determinations of their cytotoxic
activity.
Gaudichaudione I (1) was obtained as a yellow oil, [a]25D, -68.6°(c 0.07,
CHC13) with the following spectral characteristics: IR (KBr) v max cm': 3450
(hydroxyl group), 1758 (an unconjugated carbonyl group), 1685 (an a.~-unsatu-
rated carbonyl group), 1650 ( a chelated ortho-hydroxyl carbonyl group); UV (EtOH)

453
454 XIAO-HUA WU et al.

Amax (log s): 208 (4.30), 332 sh (3.50), and 358 nm (3.59); HR EIMS [M]+· 578.6638,
calcd. for C33H3809, 578.6643. The 1H NMR spectrum of 1 disclosed the presence of
five methyl groups [O 1.85 (H-15), 1.88 (H-20), 1.27 (H-25), 1.28 (H-29), and 1.78 (H-
30)], six methylenegroups [O 2.85 (H-11), 2.92 (H-11'), 5.04 (H-14), 4.99 (H-14'),
2.70 (H-16), 3.10 (H-16'), 4.85 (H-19), 4.88 (H-19'), 2.57 (H-21), 2.73 (H-21'), 1.44
(H-26), and 2.35 (H-26')], among which H-14/H-14' and H-19/H-19' were two pairs
of geminal olefinic protons. The presence of four methine protons [O 3.53 (H-7), 4.29
(H-12, connected to an oxygenated carbon), 4.27 (H-17, connected to an oxygenated
carbon), and 2.60 (H-27)] was supported by the 'H NMR spectrum of 1. The 1H NMR
spectrum of 1 furtherrevealed two olefinic protons [O 7 .58 (H-8) and 6.45 (H-22)], one
aldehyde proton [09.17 (H-24)], and one chelated hydroxyl proton [o 13.1 (OH-Cl)].
The 13C NMR and DEPT spectra of 1 indicated the presence of one aldehyde carbony I
[O 194.8 (C-24)], two ketone carbonyls [O 203.3 (C-6) and 179.0 (C-9)], three
oxygenated aromatic carbons [O 161.6 (C-1), 165.7 (C-3), and 157.2 (C-4a)], three
carbonated aromatic carbons [o 107.3 (C-2), 105.3 (C-4), and 100.5 (C-9a)], eight
olefinic carbons [o 135.7 (C-8), 133.6 (C-8a), 146.8 (C-13), 110.5 (C-14), 146.8 (C-
18), 110.9 (C-19), 147.3 (C-22), and 140.0 (C-23)], five oxygenated sp3 carbons,
which included two methine carbons (0 76.8 (C-12) and 76.0 (C-17)] and three qua-
ternary ones [o 83.7 (C-5), 90.6 (C-lOa), and 84.1 (C-28)], two methine carbons [o
46.9 (C-7) and49. l (C-27)], four methylene carbons [028.6 (C-11), 30.1 (C-16), 29.2
(C-21), and 25.1 (C-26)], and five methyl carbons [o 18.6 (C-15), 18.1 (C-20), 8.7 (C-
25), 28.8 (C-29), and 30.7 (C-30)].
 TWO MINOR, CYTOTOXIC CAGED XANTHONOIDS FROM GARCIN/A GAUD/CHAUD// 455

Table 1. 'H NMR data• for compounds 1, 2 and 3

Proton(s) 1 2 3

7 3.53b (dd, 4.6, 6.9)c 3.53 (dd, 4.5, 6.9) 3.52 (dd, 4.7, 6.8)
8 7.58 (d, 6.9) 7.58 (d, 6.9) 7.55 (d, 6.8)
11 2.85 (dd, 9.4, 13.8) 2.85 (dd, 10.0, 14.3) 2.66 (dd, 9.2, 15.0)
2.92 (dd, 3.4, 13.8) 2.93 (m) 3.11 (dd, 7.9, 15.0)
"'1"
,.....,
0 12 4.29 (m) 4.23 (dd, 10.0, 2.5) 4.28 (m)
N
•<l)... 14 5.04 (br s) 4.92 (br s) 5.04 (br s)
.n
s 4.99 (br s) 4.97 (br s) 4.88 (br s)
<l)
>
0
15 1.85 (3H, s) 1.83 (3H, s) 1.85 (3H, s)
z
V)
16 2.70 (dd, 9.4, 14.5) 2.93 (2H, m) 3.24 (dd, 5.4, 14.8)
,.....,
l'--
3.10 (br d, 14.7) 3.37 (dd, 7.6, 14.8)
N
~
17 4.27 (m) 4.38 (dd, 6.7, 3.4) 5.10 (m)
0
•.o:s.. 19 4.85 (br s) 4.86 (2H, br s) 1.74 (3H, s)
~ CJ)
4.88 (br s)
<l)
·,::o:s 20 1.88 (3H, s) 1.83 (3H, s) 1.66 (3H, s)
•...
.n 21 2.57 (dd, 7.8, 16.0) 2.63 (dd, 7.1, 15.9) 2.57 (dd, 7.8, 16.2)
;.::i
bl) 2.73 (dd, 7.2, 16.0) 2.76 (dd, 7.4, 15.9) 2.76 (dd, 6.7, 16.2)
i:::
0 22 6.45 (m) 6.34 (t, 7.1, 7.7) 6.45 (m)
~
bl) 24 9.17 (s) 9.20 (s) 9.15 (s)
i:::
0
:r: 25 1.27 (3H, br s) 1.24 (3H, s) 1.26 (3H, s)
'+-<
0 26 1.44 (dd, 9.6, 13.5) 1.42 (dd, 9.4, 13.5) 1.43 (dd, 9.4, 13.5)
>-.
.-;:: 2.35 (dd, 4.6, 13.5) 2.35 (dd, 4.5, 13.5) 2.35 (dd, 4.7, 13.5)
•<l)...
CJ)

27 2.60 (d, 9.5) 2.55 (d, 9.4) 2.59 (d, 9.4)

>
·a 29 1.28 (3H, s) 1.28 (3H, s) 1.31 (3H, s)
2 30 1.78 (3H, s) 1.66 (3H, s) 1.74 (3H, s)
>-.
.n Cl-OH 13.1 (s) 12.96 (s) 12.83 (s)
"O
<l)
"O
ee
0 a Measured in CDC13 (400 MHz for 1, 300 MHz for 2). b Chemical shifts ( o) in ppm.
'i=
;:::
0
cOne proton unless otherwise stated; s: singlet, d: doublet, m: multiplet; coupling
Ci constants in Hz in parentheses.

A comparison of the 1H NMR and 13C NMR spectra of 1 was made with those of
gaudichaudione B (3), which structure was determined by HMBC, HMQC,
NOESY, and H-H COSY. The only difference between these two natural products
was that the group at C-4 in 3 was 3-methylbut-2-enyl while for 1, the substituent
456 XIAO-HUA WU et al.

Table 2. 13C NMRa chemical shifts for compounds 1, 2 and 3

C# 1 2 3 C# 1 2 3

1 161.6 Sb 161.9 s 160.9 s 15 18.6q 18.6 q 18.5 q

2 107.3 s 107.0 s 106.9 s 16 30.1 t 29.3 t 22.3 t
3 165.7 s 165.5 s 165.4 s 17 76.0 d 76.5 d 122.2 d
4 105.3 s 105.0 s 108.4 s 18 146.8 s 146.3 s 132.5 s
4a 157.2 s 156.8 s 156.4 s 19 110.9 t 112.0 t 18.l q
5 83.7 s 83.4 s 83.7 s 20 18.1 q 17.4 q 25.7 q
6 203.3 s 203.0 s 203.4 s 21 29.2 t 29.0 t 29.1 t
7 46.9 d 46.9 d 46.9 d 22 147.3 d 147.0 d 146.9 d
8 135.7 d 135.9 d 135.4 d 23 140.0 s 140.3 s 139.9 s
Sa 133.6 s 133.3 s 133.8 s 24 194.8 d 194.4 d 194.8 d
9 179.0 s 178.9 s 178.8 s 25 8.7 q 8.8 q 8.7 q
9a 100.5 s 100.5 s 100.4 s 26 25.1 t 25.2 t 25.1 t
lOa 90.6 s 90.8 s 90.5 s 27 49.1 d 49.2 d 49.1 d
11 28.6 t 28.3 t 28.4 t 28 84.1 s 83.9 s 84.0 s
12 76.8 d 77.0d 77.5 d 29 28.8 q 28.7 q 28.8 q
13 146.8 s 146.7 s 146.6 s 30 30.7 q 30.9 q 29.9 q
14 110.5 t 110.3 t 110.5 t

aChemical shifts(~) in ppm (in CDC13, 500 MHz). b Multiplicity determined by

DEPT.

at C-4 was 2-hydroxy-3-methylbut-3-enyl. Gaudichaudione I thus had the struc-

ture assigned as 1.
Gaudichaudione J (2), also a new compound, [a.]250, -165.0° (c 0.08,
CDC13), was isolated as a yellow oil, with the following spectral characteristics:
IR (K.Br) vmax cm:': 3445 (hydroxyl group), 1754 (an unconjugated carbonyl
group), 1690 (an cx.,~-unsaturated carbonyl group), 1648 (a chelated ortho-
hydroxyl carbonyl group); UV (EtOH) A.max (log E): 224 (4.13), 336 sh (3.65), and
356 nm (3.73); HR EIMS [M]•·, 578.6652, calcd. for C33H3809 578.6643,
indicating that 2 was an isomer of 1, since both had the same molecular formula.
The 1H NMR and 13C NMR spectra of 2 were almost (but not exactly) identical
to those of 1, except for some minor differences. The same four sets of proton-
coupled systems found in 1, viz. two pairs of 2-hydroxy-3-methylbut-3-enyl, the
3-formylbut-2-enyl group, and the CHCH2CHCH=C system, were also present in
2, and were attached to the same carbon positions as in compound 1. Since 2 was
structurally similar to 1, but exhibited only subtle differences in the 1H and 13C
 TWO MINOR, CYTOTOXIC CAGED XANTHONOIDS FROM GARCIN!A GAUD!CHAUD!l 457

NMR spectra (Tables 1 and 2) and could also be separated from 1, it was therefore con-
cluded that compound 2 (gaudichaudione J) was a diastereomer of compound 1. The
absolute configurations of these compounds remain unknown.

EXPERIMENTAL
General Experimental Procedures. Optical rotations were measured in CHC13
solution at 25 °C using a Digital Polarimeter (JASCO, DIP-1000). UV spectra were
recorded on a Hewlett Packard 8452A diode array spectrometer. IR spectra were
recorded on a Bio-Rad FfIR spectrometer. EIMS were run on a Micromass VG 7035
mass spectrometer at 70 eV. NMR spectra were recorded by Bruker ACF 300 [300/400
MHz (1H)], and AMX 500 [500 MHz 125 MHz (13C)]
Plant Material. The leaves of Garcinia gaudichaudii were collected from Mt.
Tawai, Kinabatangan, Sabah, Malaysia in 1996 and identified by J. T. Pereira and L.
Madani. A voucher specimen (SAN 135145) was deposited at the herbarium of the
Forest Research Centre, Sepilok, Sandakan, Sabah, Malaysia.
Extraction and Isolation. The dried and powdered leaves ( 1 kg)were extracted
first with hexane (24 h, 5x6 L), then with CHC13 (24 h, 5x6 L), and finally with acetone
(24 h, 5x6 L) in a Soxhlet apparatus. The hexane and CHC13 extracts were evaporated
to dryness under vacuum to yield residues of 14.1 g and 30 g respectively. The CHCl3
extract was chromatographed on silica gel (Merck 9385, 1800 g) eluting with hexane,
with a gradient of ethyl acetate to 100%, followed by CHCl/MeOH ( 10: 1 to 1: 1 ). The
relatively less polar compounds comprised fifteen caged tetraprenylated xanthonoids
(gaudichaudiones A-H, gaudichaudiic acids A-E, morellic acid, and forbesione)
reported earlier- 6 while a mixture of two minor relatively polar xanthonoids eluted
much later. Preparative HPLC (RP18, 100% water with 1 % HCOOH to 100% MeCN)
separated gaudichaudiones I (1) and J (2), both - 1 mg or -0.0001 % , with 1 eluting first.
°,
Gaudichaudione I (1): yellow oil; [a]25 -68.6°(c 0.07, CHC13}; UV(EtOH} Amax
nmtlog s): 208 (4.30), 332 sh(3.50),and358 nm(3.59); IR(KBr}vmax cm:': 3450, 1758,
1685, 1650; 1H NMR (400 MHz, CDC13) and 13C NMR (125 MHz, CDC13): see Tables
1 and 2; EIMS m/z (rel. int.}: 577 [M-1]+ (40} 425 (100); HR EIMS: m/z 578.6638
(calcd. for C33H3809, 578.6643).
Gaudichaudione J (2): yellow oil; [cx]2s°, -165.0°(c 0.08, CHC13); UV (EtOH)
A.max (log E): 224 (4.13), 336 sh (3.65), and 356 (3.73) nm; IR (KBr) v max cm': 3445,
1754, 1690, 1648; 1H NMR (300 MHz, CDCl3) and 13C NMR (125 MHz, CDC13): see
Tables 1 and 2; EIMS m/z (rel. int.): 577 [M-1]+ (40) 425 (100); HR EIMS: m/z
p
578.6652 (calcd. for C33H3 9, 578.6643).
Bioassays.6-8 The following cell lines were used: P-388 (mouse leukemia} and
WEHI-1640 (mouse fibrosarcoma). The ED50 of 1 and 2 to P-338 were 0.90 and 1.06,
and to WEHI-1640 were 5.46 and 6.05 µg/mL, respectively.
 458 XIAO-HUA WU et al.

ACKNOWLEDGEMENT.
We thank the National University of Singapore for financial support and
research scholarships to XHW and SGC (now at CNPR, IMCB, NUS).

REFERENCES

1. G. Kartha, G. N. Ramachandran, H.B. Bhat, P. Madhavan Nair, V. K. V. Rapha-

van, and K. Venkataraman (1963) Tetrahedron Lett. 1, 459-472.
'1"
,.....,
0
2. L. J. Lin, L. Z. Lin, J.M. Pezzuto and G. A. Cordell (1993) Magn. Reson. Chem.
N
.<l)... 31(4), 340-347.
.n 3. J. Asano, K. Chiba, M. Tada, and T. Yoshii ( 1996) Phytochemistry 41(3), 815-820.
s
<l)
4. Y. W. Leong, L. J. Harrison, G. J. Bennett and H. T. W. Tan (1996) Chem. Research
>
0
z
V)
(S), 392-393.
,....., 5. S. G. Cao, X. H. Wu, K. Y. Sim, B. K. H. Tan, J. T. Pereira, W. H. Wong, N. F.
l'--
N Hew and S. H. Goh (1998) Tetrahedron Lett. 39, 3353-3356.
~
0 6. S. G. Cao, Valerie H. L. Sng, X. H. Wu, K. Y. Sim, B. K. H. Tan, J. T. Pereira and
.•....
~
o:s S. H. Goh (1998) Tetrahedron 54, 10915-10924.
CJ)
<l)
·,::o:s 7. T. Mosmann (1983) J. Immun. Methods 65, 55-63.
.... 8. R. I. Geran, N. H. Greenberg, M. M. MacDonald, A. M. Schumacher and B. J.
.n
;.::i Abbott (1972) J. Cancer Chemotherapy Reports, Part 3 3(2), 1-17, 59-61.
bl)
i:::
0
~
bl)
i:::
0
:r:
'+-<
0
>-.
.-;::
.<l)...
CJ)

>
·a
2
>-.
.n
"O
<l)
"O
ee
0
"i=
;:::
0
Ci