Triterpenoid Dammarans From The Bark of Aglaia Smithii (Meliaceae) And Their Toxic Activity Against Artemia Salina And Cytotoxic Activity Against Murine Leukimia Cells P-388.

TRITERPENOID DAMMARANS FROM THE BARK OF AGLAIA SMITHII
(MELIACEAE) AND THEIR TOXIC ACTIVITY AGAINST ARTEMIA SALINA
AND CYTOTOXIC ACTIVITY AGAINST MURINE LEUKEMIA CELLS P-388

Desi Harneti Putri Huspa a), Unang Supratman a), Tati Herlina a), Syafruddin b)
a)

Jurusan Kimia FMIPA UNPAD, b) Lembaga Eijkman Jakarta

* Corresponding author, tel/fax : 022-7794391, email: d_harneti@yahoo.com
ABSTRACT

Aglaia smithii is a higher plant belonging to Meliaceae family and widely
distributed in South East Asia. Plants of Meliaceae family were known as a
source of cytotoxic (anticancer) substances. The methanolic extract from the
dried bark of A. smithii was concentrated and extracted successively with nhexane, methylene chloride and ethyl acetate. By using Brine Shrimp Lethality
Test (BSLT) to follow the separation (bioassay guided isolation), the compounds
of n-hexane extract was separated by combination of column and thin layer
chromatography to yield two toxic compounds. The chemical structure of active
compounds were determined by spectroscopic data and compared with those
spectra data reported previously were identified as eichlerianic acid (1) and

aglinin A (2). The toxic activity of compounds (1) and (2) against brine shrimp (A.
salina) were evaluated by Meyer method and showed activity with LC 50 24,7and
36,4; µg/mL, respectively. The cytotoxic activity of compounds (1) and (2) against
murine leukemia cells P-388 were evaluated by Alley Method and showed weak
activity with IC50 34 and 42 µg/mL, respectively.

Keywords: Aglaia smithii; Meliaceae; brine shrimp lethality test; murine
cell P-388; triterpenoid dammaran

INTRODUCTION

leukemia

Indonesia, we have isolated from the

Plants belonging to the genus

bark

of

Aglaia

smithii

the

known

Aglaia are a rich source of tetracyclic

dammaran triterpenoids eichlerianic acid

triterpenoids

(1) and aglinin A (2).

of

the

cycloartane,

dammarane and tirucallane series [1]

Aglaia smithii is a higher plant

In continuation of our work on plants

belonging

belonging to this genus collected in

widely distributed in South East Asia.

to

Meliaceae

family and

Plants

of

Meliaceae

family

were

P-388 were evaluated by Alley Method

known

as

a source of cytotoxic

and showed weak activity with IC50 34

(anticancer) substances [2-4] . The

and 42 µg/mL, respectively.

methanol extract from bark of A.
EXPERIMENTAL SECTION
smithii

showed

significant

activity
Materials

against brine shrimp (Artemia salina)
Bark material of Aglaia smithii Corr.
and murine leukemia cells P-388. This
was

collected

in

Bogor

Botanical

plant has not yet been subjected to
Garden West Java
any

phytochemical

or

Indonesia, during

biological

October

2006.

The

specimen

was

investigation. The methanolic extract
identified

and

deposited

at

the

from the dried bark of A. smithii was
Herbarium
concentrated

and

extracted

successively

with

n-hexane,

Bogoriense

West

Java

Indonesia.
Instrumentation
methylene chloride and ethyl acetate.
Rotavapor R-200 Buchi with vacum Vac
By using Brine Shrimp Lethality Test
V-500
(BSLT)

to

follow

the

Buchi.

Fischer-Johns

Melting

separation
Point Apparatus. Spektrofotometer FTIR

(bioassay

guided

isolation),

the
Shimadzu 8400 and FTIR Spectrum

compounds of n-hexane extract was
One Perkin Elmer. Spektrofotometer
separated by combination of column
NMR (Nuclear Magnetic Resonance)
and thin layer chromatography to yield
JEOL
two

toxic

compounds.

The

JNM

ECA-500.

Mariner

toxic
Biospectrometry, Hitachi L 6200, sistem

activity of compounds (1) and (2)
ESI

(Electrospray Ionisation), positive

against brine shrimp (A. salina) were
ion mode and Shimadzu LCMS solution,
evaluated

by

Meyer method

and
negative ion mode. Bruker SMART

showed activity with LC50

24,7 and
APEX Diffractometer.

36,4;

µg/mL,

respectively.

The

cytotoxic activity of compounds (1)
Procedure
and (2) against murine leukemia cells

Extraction and isolation

The 3,4 secodammaranes, eichlerianic

Dried bark of A. smithii (3 kg)
were

extracted

with

from the n-hexane extract of A. smithtii

methanol at room temperature. The

bark by chromatography on silica gel.

extract (286 g) was diluted with

They have been found previously in A.

methanol

and

lawii leaves [5]. However, they have

partitioned with n-hexane. The extract

been described in other genera of the

(13

repeatedly

family Meliaceae and were identified on

chromatographed on silica gel yielding

the basis of comparison of their spectral

eiclerianic

n-

data with literature values [1,5]. They all

hexane/ethyl acetate, 7 : 3, (2) n-

possess the same 20S,24-epoxy-25-

heksana/CH2Cl2/MeOH, 1,4 : 85 : 0,1.

hydroxy chain at C-17.

:

exhaustively

acid (1) and aglinin A (2) were isolated

water

g)

was

acid

(23

(8:2

v/v)

mg)

(1)

Aglinin A (21 mg) CH2Cl2/MeOH,
MeOH : H2O, 9:1.
Bioassay

Eichlerianic acid
Eichlerianic acid showed an

[M-

The toxic activity of compounds (1)

H]- peak at m/z 473 in the LC-MS,

and (2) against brine shrimp (A.

coresponding

salina)

C30H50O4.

were evaluated by Meyer

to

molecular

formula

method and showed activity with LC50

Recent detailed NMR studies have

24,7and 36,4; µg/mL, respectively.

unambiguously demonstrated that the

The cytotoxic activity of compounds

24R and 24S isomer can be easily

(1) and (2) against murine leukemia

distinguished by the resonances of C-24

cells P-388 were evaluated by Alley

(δC 83.2 for the R-isomer and δC 86.5

Method and showed weak activity with

for the S-isomer) [5]. The chemical shifts

IC50 34 and 42 µg/mL, respectively.

and coupling patterns of the H-24 differ
also from each other (δH3.7, J =7 and 7

RESULTS AND DISCUSSION

Hz and δH 3.6, J = 10 and 5.5 Hz,
respectively)

(Table

1).

Position

carboxyl group at C-3 and double

the signal of the C-3 carbonyl appeared

bond at C-4 and C-28 with HMBC

at δ 178.5.

correlation H-28/C-5, C-29; H-29/C-

These data as well as all the signals of

28,C-5,C-4; H-2/C-3 indicated ring A is

the dammarane fused ring moiety were

open.

similar

The IR showed the absorption of a

3,4secodammaranes eichleirianic acid

carbonyl group at 1708 cm-1.

previously isolated. The side chain

In the

13

C NMR the signal of the

carbonyl

appears

at

δ

to

the

ones

of

the

signals, however, differed from those of

179.7

the latter. In the NMR spectra, the

suggesting a carboxylic acid residue.

typical signals of the C-24 oxymethine

The1H NMR exhibited the signal of

were absent. Instead, a signal of a C-

seven tertiary methyls between δ 0.8

OH at low field (δ 108.7) was observed

and 1.3, ten methylene, four methin

in the

and of an oxymethineat δ 3.63 typical

tetrahydrofuran ring was replaced by a

of the

five membered hemiacetal ring. The 1D

H-24 in 20S,24S-epoxy-25-

hydroxydammaranes.
signals were identic

All

the

13C

to the ones of

13

C NMR suggesting that the

and 2D (COSY, HMQC, HMBC) NMR
data

were

in

agreement

with

the

eichlerianic acid.

structure depicted in Figure 1, especially

Aglinin A

the diagnostic HMBC correlations Me-

Aglinin A showed an [M-H]- peak

21/C-17, C-20,C-22 and Me-26, Me-

m/z

LC-MS,

27/C-24. C-20 was assigned the S

coresponding to molecular formula

configuration similar to most of the

C30H50O5.

dammarane

at

The

489,3

IR

in

spectrum

the

showed

the

particularly

those isolated from the Aglaia genus

absorption of a carboxyl at 1710 cm-1.
In the

triterpenes

13

C NMR spectrum (Table 2),
CONCLUSION

Two toxic 3,4-secodammarane
triterpenes, eichlerianic acid (1) and

1. Roux, D., T. Martin., T. Adeline.,
T. Sevenet., H. Hadi., and Pais,
M. 1998. Foveolins A dan B,

aglinin A (2), were isolated from the
stem

bark of Aglaia smithii with

bioactivity

(Brine

Shrimp

Lethality

dammarane

triterpenes

from

Aglaia foveolata. Phytochemistry
49(6): 1745-1748.
2. Omar S., Zhang, J., Kinnon,

Test)

guided

chromatografic

M.S., Leaman, D., Arnason, J.T.,
and

fractionation .

Philogen,

B.J.R.

Traditionally-used

2003.

antimalarials

from Meliaceae. Current Top
2. The toxic activity of compounds (1)
and (2) against brine shrimp (A.
salina) showed activity with LC50 24,7

Medicinal Chemistry 3(2): 133139.
3. Nugroho, B. W., Edrada, R.A.,
Wray, V., Witte L., Bringmann,

and 36,4; µg/mL, respectively. The
cytotoxic activity of compounds (1)
and (2) against murine leukemia cells

G., Gehling, M., and Proksch, P.
1999.

An

rocaglamida

insectisidal
derivates

and

related compounds from Aglaia
P-388 showed weak activity with IC50

odorata

34 and 42 µg/mL, respectively.

Phytochemistry 51: 367-376.

(Meliaceae).

4. Nakatani,

M.,

Abdelgalell,

S.A.M., Sand, M.M.G., Huang,
ACKNOWLEDGEMENTS

R.C., Doe, N., and Iwagawa, T.

We thank for The Ministry of National
Education,

Directorate

of

Higher

2004.

Phragmalin

Limonoids

from

Chukrasia

tabularus.

Phytochemistry 65: 2833-2841.
Education, Republik Indonesia, for
financial support (STRANAS Project
2010) and LIPI Serpong for NMR
measurement.

5. Mohammad,

M.-T.,

Leroy, E., Tempete, C., Sevenet,
T., Awang, K., and Pais, M.
1999. Dammarane Triterpenes
and

REFERENCES

K.,Martin,

Pregnane

Steroids

from

Aglaia lawii and A. tomentosa.
Phytochemistry 51: 1031-1037.

Table 1

13

C (125 MHz) and 1H NMR (500 MHz) data for eichlerianic acid (CDCl3)
Posisi C
1
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
28
29
30



C-NMR.
c (mult.)
34,44 (t)
28,14 (t)
178,39 (s)
147,70 (s)
49,98 (d)
25,00 (t)
34,06 (t)
40,22 (s)
41,36 (d)
39,25 (s)
22,52 (t)
24,76 (t)
43,07 (d)
50,55 (s)
31,65 (t)
27,08 (t)
49,90 (d)
16,50 (q)
20,39 (q)
86,73 (s)
27,40 (q)
34,87 (t)
26,51 (t)
86,52 (d)
70,44 (s)
28,08 (q)
24,23 (q)
113,63 (t)
23,38 (q)
15,52 (q)

1

H-NMR
H, (H, mult., J (Hz))
1,43 (2H,m)
2,20; 2,39 (2H,m)
1,96 (1H,m)
1,87 (2H,m)
1,58 (2H, m)
1,52 (1H,m)
1,43 (2H, m)
1,38 (2H, m)
1,64 (1H, m)
1,46 (2H, m)
1,34 (2H, m)
1,86 (1H, m)
0,89 (3H, s)
0,86 (3H, s)
1,14 (3H, s)
1,58 (2H, m)
1,62 (2H, m)
3,63 (1H, dd, J = 5,5 dan 9,8)
1,19 (3H, s)
1,11 (3H, s)
4,66; 4,85 (2H, brs)
1,73 (3H, s)
1,01 (3H, s)

HMBC
(HC)
C-3

C-5, C-9

C-7,C-8, C-14
C-1, C-5,C-9, C-10
C-17,C-20,C-22

C-27,C-25,C-24
C-26, C-25,C-24
C-29,C-5
C-5,C-28,C-4
C-8,C-13,C-14

Table 2

13

C (125 MHz) and 1H NMR (500 MHz) data for aglinin A (CDCl3)
Posisi C
1
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
28
29
30



C-NMR.
c (mult.)
34,08 (t)
28,22 (t)
178,54 (s)
147,75 (s)
50,95 (d)
27,98 (t)
31,24 (t)
39,17 (s)
43,33 (d)
40,24 (s)
22,20 (t)
27,58 (t)
41,42 (d)
50,51 (s)
34,33 (t)
26,35 (t)
51,05 (d)
20,37 (q)
15,52 (q)
89,15 (s)
24,71 (q)
31,78 (t)
31,82 (t)
108,74 (s)
74,76 (s)
24,78 (q)
24,32 (q)
113,67 (t)
23,36 (q)
16,60 (q)

1

H-NMR
H, (H, mult., J (Hz))
1,19; 1,23 (2H,m)
2,15; 1,40 (2H,m)
1,95 (1H,m)
1,82; 1,96 (2H,m)
1,08 (2H, m)
1,52 (1H,m)
1,38; 1,43 (2H, m)
1,82; 1,96 (2H, m)
1,49 (1H, m)
1,60 (2H, m)
1,36 (2H, m)
1,99 (1H, m)
0,88 (3H, s)
1,00 (3H, s)
1,13 (3H, s)
1,07; 1,84 (2H, m)
1,82 (2H, m)
1,28 (3H, s)
1,25 (3H, s)
4,65; 4,85 (2H, brs)
1,71 (3H, s)
0,84 (3H, s)

HMBC

C-1, C-3

C-8, C-14
C-21
C-7,C-8, C-9,C-14
C-1, C-5, C-10
C-17,C-20,C-22
C-20

C-24, C-25
C-24, C-25, C-27
C-29,C-5
C-5,C-28,C-4
C-8,C-14,C-15

ARTIKEL ILMIAH
HIBAH PENELITIAN STRATEGI NASIONAL

TRITERPENOID YANG BERSIFAT ANTIMALARIA DARI
TUMBUHAN MELIACEAE INDONESIA

Oleh :

Dr. Desi Harneti Putri Huspa
Prof. Dr. Unang Supratman
dr. Syafruddin, Ph.D
Dr. Tati Herlina

DIBIAYAI OLEH :
DIREKTORAT JENDRAL PENDIDIKAN TINGGI,
KEMENTRIAN PENDIDIKAN NASIONAL
SESUAI DENGAN SURAT PERJANJIAN PELAKSANAAN
HIBAH PENELITIAN
NOMOR : 153/SP2H/PP/DP2M/III/2010
TANGGAL 1 MARET 2010

DEPARTEMEN PENDIDIKAN NASIONAL
UNIVERSITAS PADJADJARAN
FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM
JANUARI, 2011