Journal of Applied Pharmaceutical Science Vol. 5 (01), pp.

110-113, January, 2015

Available online at http://www.japsonline.com
DOI: 10.7324/JAPS.2015.50119
ISSN 2231-3354

Short Communication

Piceatanol: Anti-Cancer Compound From Gewang Seed Extract

Leny Heliawati*1.2, Agus Kardinan3,Tri Mayanti1, Roekmi-ati Tjokronegoro1
1
Graduate School, Padjadjaran University, Bandung, West Java, Indonesia. 2Pakuan University, Bogor, West Java, Indonesia. 3Indonesian Spice and
Medicinal Crops Research Institute, Indonesia

ARTICLE INFO ABSTRACT

Article history:
Received on: 22/09/2014 Piceatannol (Compound 1), brownish white solids compound, is a stilbene compound has been isolated from
Revised on: 11/11/2014 methanol seeds extract of Corypha utan Lamk. Isolation and purification conducted by chromatographic
Accepted on: 17/01/2015 methods. Structure elucidation deduced on the basis of spectroscopic data (UV spectrometer, FTIR, NMR and
Available online: 30/01/2015 HRMS). MTT assay method of cytotoxicity activity showed that Compound 1 has a very strong cytotoxic
activity against Murine leukemia P-388 cell lines with IC50 value 1.56 ppm.
Key words:
Corypha utan Lamk, Murine
leukemia P-388, Cytotoxic,
Chromatography,
Piceatannol.

INTRODUCTION ppm (inactive). The purpose of this study is to isolate anticancer

active compound from the seed of Corypha utan Lamk.
Research and development of drugs is a very important
part of health development, requires the development of new
compounds as ingredients of medicines. Natural products are MATERIALS AND METHODS
important sources of new structures, especially for the discovery of
Extraction and Isolation
compounds are efficacious drugs. Presently, natural products drug 3.9 kg of Corypha utan Lamk fresh fruit collected from
development focused on the search and analysis of the new Buat So’e area, District of Timor Tengah Selatan, Nusa Tenggara
compounds that might be useful as a medicine. The selection of
Timur Province, Indonesia. Separate the seed from the flesh,
suitable plants is an important and decisive step, can be done crushed, and then macerated with 3 liters of methanol for 3 days.
several ways, among others, the use of traditional, chemical Liquid methanol extract filtered and concentrated using vacuum
constituents, toxicity, random selection of a combination of several rotary evaporator at ± 40 °C temperature. 20 g methanol extract
criteria (Gudrun et al., 2010). Corypha utan Lamk. is a type of fractionated using vacuum liquid chromatography (silica gel GF254)
palm plant that grows wild in the savanna of East Nusa Tenggara with n-hex-EtOAc 1:1; 4:6; 3:7; 2:8 and EtOAc as mobile to obtain
(NTT), used as fish poison by the people of Timor Island. Murine
five fractions (A1-A5). Fraction A5 further chromatographed over
leukemia P-388 is one of the tumor cells type that serve as a silica gel column, eluted successively with n-hex-EtOAc (2:8), to
cytotoxic test protocol by NCI (National Cancer Institute) give 50 mg of brownish white solid compound (Compound 1).
America. Test results using these cells are often used as the basis
of the tests in order to obtain further compounds or candidate
Anticancer Activity Test Against murine leukemia P-388
cancer models (Hoetetman and Hamburger, 1991). A pure
The principle of the measurement of the cytotoxic
compound categorized as anticancer active compound if it has IC50
properties of murine leukemia cancer cells P-388 are as follows: the
value <2 ppm (very active), IC50 2-4 ppm (active) and IC50 > 4 .

.
activity of the compounds and Antonin E (positive control) is
* Corresponding Author expressed by the IC50 which is sample concentration or comparison
Leny Heliawati, e-mail: [email protected] is needed to inhibit 50% tumor cells Murine leukemia P-388 cell

© 2015 Leny Heliawati et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-ShareAlike
Unported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
 Heliawati et al. / Journal of Applied Pharmaceutical Science 5 (01); 2015: 110-113 111

13
line through MTT reagent staining, which was observed with a C-NMR spectrum show six aromatic carbon at δC 102.5; 105.8
micro plate reader at 540 nm. Approximately 3 x 104 cell cm-3 of (2C); 113.7; 116.3; 120.3; 126.7; 129.5; and two olefinic carbon at
P-388 Murine leukemia cells were plated in 96-well culture dishes, δC 146.02 (2 C), 2 carbon chemistry shift value at δC = 159.08 (2C)
and incubated for 24 h. various concentrations of the samples were and two aromatic carbon quarterner at δC 130.9 and 141.17 (Figure
added. Six desirable sample concentrations were prepared using 4). Mass spectroscopy analysis showed that compound 1 has a
PBS (phosphoric buffer solution, pH = 7.30-7.65), except control. molecular weight (m/z) 245 and molecular formula C14H13O4.
After 48 h incubation, the test was stop by adding MTT reagent [3- Further identification of compound 1 was determined by HMQC
(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide]. and HMBC (Figure 5), showed that protons at δH 7.01 correlated
Incubation continue for next 4 h before the addition of MTT stop with δC 120.3 (C-6 ') and δC 145.9 (C-4'). The opposite correlation
solution containing sodium dodecyl sulphate (SDS), the incubation also showed between δH 6.82 with δC 113.7 signal (C-2'). Proton at
continue for next 24 h. optical density measured using microplate δH 6.82 showed correlation with the two aryl carbon δC 145.9 (C-4
reader at 540 nm. IC50 value calculated using extrapolation of 50% ') and δC 146.0 (C-3'), δH 6.77 also has correlation with δC 145.9 (C
absorption lines in the positive control sample on the uptake curve -4'), δC 146.0 (C-3'), and quaternary aromatic carbon δC 130.9 (C-
against sample concentration. 1'). Proton signals δH 6.74 (C-7) had a trans coupling with δH 6.89
(C-8). HMBC spectrum also showed other correlation between δH
RESULTS AND DISCUSSION 7.01 and δH 6.77 with δC signal (C-1'), and δH 6.89 with δC (C-7)
and carbon quaternary (C-1) second aromatic ring in unit A.
Compound 1, a brownish white solid with a melting
HMQC and HMBC correlation of compound 1 shown in Table 1.
point of 226oC. UV (MeOH, λmax)(log ε) nm: 221 (tape
The relationship between proton-carbon neighbor within 2 ties and
conjugates) and 327 (tape benzene). These data indicated that in
3 ties of the HMBC spectrum of compound 1 is shown in Figure 5.
this compound under electronic transition π  π * which
Based on the 1D- and 2D-NMR data, supported with mass
characterizes a chromophore of an aromatic substitute with
spectroscopic data and compared with a reference (Brinker and
auxochrome and under bathochromic shift with the addition of
Seigler, 1991) can be concluded that compound 1 is Piceatannol.
NaOH reagent (λmax)(log ε) nm: 309 and 347, showed that
Cytotoxicity activity against Murine leukemia P-388 cell lines of
compound 1 has free OH group (Figure 1). IR spectrum showed
compound 1 has been done. Compound 1 showed strong activity
conjugation absorption bands (vmaks cm-1) for hydroxyl groups
with IC50 1.56 ppm compared to Artonin E (IC50 0.3 ppm) as
(3348), -C=C- of aromatic ring (1650) supported by =CH alkenes
positive control.
and aromatics (652, 800, and 960) (Figure 2). 1H-NMR spectrum
(Figure 3) indicate the presence of three ABX system
CONCLUSION
proton aromatic signals of A ring at δH 6.82 (1H, dd, C-6'); δH 7.01
(1H, d, C-2'), and δH 6.77 (1H, s, C-5'), three proton aromatic Anticancer active compound contained in Corypha utan
signal of B ring at δH 6.5 (2H, d), and δH 6.25 (1H, t), Lamk. seeds successfully isolated and identified as piceantannol,
and two proton signals at δH 6.74 (1H, d), and δH 6.89 (1H, d) which also has very strong cytotoxic activity against Murine
belong to a system of a typical trans vinyilic stilbenoid group. leukemia P-388 cells with IC50 values 1.56 ppm.

Fig. 1: UV spectrum of compound 1.

112 Heliawati et al. / Journal of Applied Pharmaceutical Science 5 (01); 2015: 110-113

Table. 1: 1D- and 2D-NMR Data of Compound 1 and Reference (Brinker and Seigler, 1991).
δH (int, mult, J = Hz) 500 MHZ (ppm)
No HMQC HMBC
1* (Brinker and Seigler, 1991) 1
1 141.17
2.6 6,43 (2H,d) 6,50 (2H,d) 105,84 (2C) 102.5 126.76
3.5 159,08 (2C)
4 6,15 (1H,t) 6,25 (1H,t) 102.5 105.84 159.08
7 6,73 (1H,d) 6,74 (1H,d) 126.76 105.84 130.91
8 6,89 (1H,d) 6,89 (1H,d) 129.59 126.76 141.17
1' 130.91
2' 6,73 (1H,d) 6,77 (1H,s) 116.38 130.91 145.98 146.02
3' 146.02
4' 145.98
5' 6,83 (1H,dd) 6,82 (1H,dd) 120.3 113.75 145.98 146.02
6' 6,97 (1H,d) 7,01 (1H,d) 113.75 113.75 120.3 145.98

90

%T

765.74
2852.72

723.31
75
2926.01

860.25

551.64
580.57
60

844.82

657.73
621.08
45

1188.15
1242.16

684.73
1197.79

1012.63

821.68
30
3510.45

964.41
1444.68

1114.86
1332.81
1361.74
1519.91

1346.31
3331.07

3296.35
3348.42

3313.71

15
1294.24
1600.92

1141.86

4500 4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500
7 1/cm
Fig. 2: IR spectrum of compound 1.

Fig. 3: 1H-NMR spectrum of compound 1.

 Heliawati et al. / Journal of Applied Pharmaceutical Science 5 (01); 2015: 110-113 113

Fig. 4: 13C-NMR spectrum of compound 1.

Fig. 5: HMQC and HMBC Correlations of Compound 1.

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