Four compounds were isolated from the stem barks of Anacolosa poilanei Gagnep. Theirs structures were established by spectroscopic analysis including MS and NMR. Accordingly, the isolates were identified as trichadenic acid B (1), trichadonic acid (2), amentoflavone (3) and β-sitosterol (4)

Cuong, Pham Van

Litaudon, Marc

Mai Huong, Doan Thi

Minh, Chau Van

Thanh Van, Trinh Thi

Thuy Ai, Doan Thi

Tram, Le Huyen

English

Vietnam Academy of Science and Technology: Journals Online

 124 
VIETNAM JOURNAL OF CHEMISTRY         VOL. 53(2e) 124-126                                            APRIL 2015 
DOI: 10.15625/0866-7144.2015-2e-029 
  
 
 
CONSTITUENTS FROM STEM BARKS OF ANACOLOSA POILANEI 
GAGNEP. (OLACACEAE) 
Doan Thi Thuy Ai
1,2
, Trinh Thi Thanh Van
1
, Doan Thi Mai Huong
1
, Marc Litaudon
3
,  
Le Huyen Tram
4
, Chau Van Minh
1
, Pham Van Cuong
1*
 
1
Institute of Marine Biochemistry, Vietnam Academy of Science and Technology 
2
Vietnam National Univercity of Agriculture, Trau Quy, Gia Lam, Hanoi 
3
Institute of Natural Product Chemistry, Gif Sur-Yvette, Paris France 
4
Hanoi University of Science and Technology, 1 Dai Co Viet, Hanoi, Vietnam 
Received 23 January 2015; Accepted for Publication 18 March 2015 
  
Abstract 
Four compounds were isolated from the stem barks of Anacolosa poilanei Gagnep. Theirs structures were 
established by spectroscopic analysis including MS and NMR. Accordingly, the isolates were identified as trichadenic 
acid B (1), trichadonic acid (2), amentoflavone (3) and β-sitosterol (4). 
Keywords. Anacolosa poilanei, Olacaceae, trichadenic acid B, trichadonic acid, amentoflavone. 
 
1. INTRODUCTION 
 
Anacolosa genus comprises about 22 species and 
belongs to the Olacaceae family. The plants of this 
genus are distributed in the tropical regions [1]. An 
overview in the literature revealed that only two 
species have been examined for their chemical 
contents [2, 3]. These phytochemical studies led to 
the isolation and characterization of terpenoids and 
unsaturated faty acids containing triple bonds which 
were relatively rare in nature. In our screening 
program, a stem bark extract of A. poilanei Gagnep. 
(Olacaceae) showed cytotoxicity against KB cells   
(> 50 % inhibition at 1 μg/mL). In this paper, we 
report the isolation and structural determination of 
four compounds 1-4.  
 
2. EXPERIMENTAL 
 
2.1. General experimental procedures 
 
Optical rotations were measured on a Polax-2 L 
polarimeter. Melting points were determined using a 
Buchi B-545 instrument. ESI-MS were obtained on 
an Agilent 1100 LC-MSD Trap spectrometer. The 
NMR spectra were recorded on Bruker 500.13 MHz 
spectrometer, operating at 500.13 MHz for 
1
H and 
125.76 MHz for 
13
C NMR, respectively.  
2.2. Plant material 
 
Stem barks of A. poilanei were collected in 
Sapa, Lao Cai province, Vietnam in June 2003. A 
voucher specimen (VN-1124) was deposited at the 
Institute of Ecology and Biological Resources, 
Vietnam Academy Science and Technology. 
 
2.3. Extraction and isolation 
 
Dry powdered stem barks of A. poilanei (1.18 
kg) was extracted with EtOAc (3×2.5 L). The 
EtOAc solution was concentrated under reduced 
pressure to dryness. The EtOAc extract (55.3 g) was 
subjected to column chromatography (CC) on silica 
gel eluting with n-hexane/EtOAc gradient to yield 
13 fractions. 
Fraction 6 (1.2 g) was separated by column 
chromatography (CC) on silica gel, eluting with n- 
CH2Cl2 gradient to afford 1 (15 mg). Fraction 8 (1.3 
g) was subjected to a Sephadex LH-20 CC to give 3 
subfractions. Subfraction 2 was purified by CC on 
silica gel (CH2Cl2/MeOH gradient), followed by 
recrystallization in EtOAc to afford 2 (16 mg). 
Fraction 9 was recrystallized in EtOH to yield 3 (15 
mg). Fraction 12 was purified on a silica gel CC 
(CH2Cl2/MeOH gradient), furnishing 4 (15 mg). 
Trichadenic acid B (1): white powder, m.p. 
302-304 
oC, [α]D +30 (c, 0.14, CHCl3) (literature: 
292-294 
oC), [α]D +32.2
 
(c, 0.69, pyridine). 
1
H-NMR 
 125 
VJC, Vol. 53(2e), 2015                                                                                                   Pham Van Cuong, et al. 
(500 MHz, CDCl3+CD3OD), δ (ppm): 3.26 (1H, td, 
H-3), 0.83 (3H, d, J = 7.0 Hz, H-23 ) 0.72 (3H, s, H-
24), 0.81 (3H, s, H-25), 0.90 (3H, s, H-26), 1.12 
(3H, s, H-28), 1.10 (3H, s, H-26), 0.94 (3H, s, H-30). 
13
C-NMR (125 MHz, CDCl3+CD3OD): δ (ppm): 
19.5 (C-1), 35.6 (C-2), 72.0 (C-3), 52.8 (C-4), 37.1 
(C-5), 41.1 (C-6), 18.0 (C-7), 52.8 (C-8), 37.7 (C-9), 
59.9 (C-10), 37.8 (C-11), 27.8 (C-12), 54.5 (C-13), 
39.1 (C-14), 32.9 (C-15), 35.8 (C-16), 30.5 (C-17), 
43.1 (C-18), 36.3 (C-19), 28.3 (C-20), 32.5 (C-21), 
38.0 (C-22), 9.7 (C-23), 14.0 (C-24), 18.0 (C-25), 
19.5 (C-26), 179.7 (C-27), 31.0 (C-28), 35.1 (C-29), 
30.6 (C-30). ESI-MS (negative): 457 [M-H]
-
. 
Trichadonic acid (2): white powder, m.p. 249-
252 
oC, [α]D +5.3
 
(c, 1.5, CHCl3) (literature: 248-249 
oC), [α]D +5
 
(c, 0.3, CHCl3). 
1
H-NMR (500 MHz, 
CDCl3), δ (ppm): 0.87 (3H, d, J = 7.0 Hz, H-23), 
0.72 (3H, s, H-24), 0.91 (3H, s, H-25), 1.14 (3H, s, 
H-26), 1.22 (3H, s, H-28), 1.00 (3H, s, H-29), 0.96 
(3H, s, H-30). 
13
C-NMR (125 MHz, CDCl3, δ (ppm): 
22.7 (C-1), 41,3 (C-2), 213.0 (C-3), 58.1 (C-4), 42.1 
(C-5), 41.3 (C-6), 18.5 (C-7), 53.0 (C-8), 37.6 (C-9), 
59.4 (C-10), 37.8 (C-11), 27.8 (C-12), 54.8 (C-13), 
39.2 (C-14), 33.0 (C-15), 41.0 (C-16), 30.7 (C-17), 
43.3 (C-18), 35.7 (C-19), 28.4 (C-20), 32.4 (C-21), 
36.0 (C-22), 16.8 (C-23), 14.7 (C-24), 18.4 (C-25), 
22.7 (C-26), 181.0 (C-27), 31.0 (C-28), 30.5 (C-29), 
35.4 (C-30). ESI-MS (negative): 455 [M-H]
-
. 
Amentoflavone (3): yellow powder, m.p. 260-
261 
o
C (literature: 254-256 
o
C). 
1
H-NMR (500 MHz, 
CD3OD): δ (ppm): 6.55 (1H, s, H-3), 6.17 (1H, d, J 
= 2.0 Hz, H-6), 6.39 (1H, d, J = 2.0 Hz, H-8), 7.97 
(1H, d, J = 2.5 Hz, H-2’), 7.08 (1H, d, J = 8.5 Hz, H-
5’), 7.83 (1H, dd, J = 2.0 Hz, 9.0 Hz, H-6’), 6.56 
(1H, s, H-3’’), 6.34 (1H, s, H-6”), 7.50 (2H, d, J = 
8.7 Hz, H-2’”, 6’”), 6.71 (2H, d, J = 8.7 Hz, H-3”’, 
5’”). 13C-NMR (125 MHz, CD3OD): δ (ppm): 166.2 
(C-2), 103.9 (C-3), 183.8 (C-4), 163.1 (C-5), 100.2 
(C-6), 163.1 (C-7), 95.2 (C-8), 159.3 (C-9), 105.3 
(C-10), 123.2 (C-1’), 132.8 (C-2’), 121.8 (C-3’), 
161.2 (C-4’), 117.7 (C-5’), 129.3 (C-6’), 165.9 (C-
2’’), 103.3 (C-3’’), 184.2 (C-4’’), 162.5 (C-5’’), 
100.2 (C-6’’), 164.0 (C-7’’), 100.3 (C-8’’), 156.5 
(C-9’’), 105.6 (C-10’’), 123.1 (C-1”’), 129.3 (C-
2’”), 115.8 (C-3’”), 162.5 (C-4’”), 115.8 (C-5’”), 
129.3 (C-6’”). 
β-Sitosterol (4): white powder, m.p. 140-142 
o
C, 
1
H-NMR (500 MHz, CDCl3), δ (ppm): 3.52 (1H, 
m, H-3), 5.35 (1H, dd, J = 3.0 Hz, H-6), 0.68 (3H, s, 
H-18), 1.01 (3H, s, H-19), 0.92 (3H, d, J = 7.0 Hz, 
H-21), 0.82 (3H, d, J = 7.0 Hz, H-26), 0.83 (3H, d, J 
= 7.0 Hz, H-27), 0.85 (3H, t, J = 7.0 Hz, H-29). 
13
C-
NMR (125 MHz, CDCl3, δ (ppm): 37.3 (C-1), 31.7 
(C-2), 71.8 (C-3), 42.3 (C-4), 140.8 (C-5), 121.7 (C-
6), 31.9 (C-7), 31.9 (C-8), 50.2 (C-9), 36.5 (C-10), 
21.1 (C-11), 39.8 (C-12), 42.4 (C-13), 56.8 (C-14), 
24.3 (C-15), 28.3 (C-16), 56.1 (C-17), 12.0 (C-18), 
19.8 (C-19), 36.2 (C-20), 18.8 (C-21), 34.0 (C-22), 
26.1 (C-23), 45.9 (C-24), 29.2 (C-25), 19.1 (C-26), 
19.4 (C-27), 23.1 (C-28), 12.0 (C-29). 
 
3. RESULTS AND DISCUSSION 
 
Compound 1 was isolated as white powder and 
optically active, [α]D +30 (c, 0.14, CHCl3). Its ESI-
MS (negative) showed pseudo-molecular ion at m/z 
455 [M-H]
-
.
 
The 
1
H-NMR spectrum of 1 indicated 
signals of a doublet methyl at δH 0.83 (J = 7.0 Hz, 
CH3-23), six singlet meyhyls at δH 0.72 (CH3-24), 
0.81 (CH3-25), 0.90 (CH3-26), 1.12 (CH3-28), 1.10 
(CH3-29), 0.94 (CH3-30), an oxymethine proton at 
δH 3.26, and a complex set of overlapping signals at 
aliphatic region. Analyses of the 
13
C-NMR and 
DEPT spectra with the aid of HSQC spectrum of 1 
revealed the signals of 30 carbons, including a 
carboxylic group, seven methyls, eleven methylenes, 
five methines and six quaternary carbons. This 
observation strongly suggested that compound 1 
should be a triterpene belonging to friedelane 
skeleton. The observation of only seven methyl 
groups and the presence of a carboxylic carbon 
suggested one methyl group being oxidized into a 
carboxylic functionality. Analyses of 2D NMR 
spectra of 1 confirmed the structure of 1 in which 
the methyl group CH3-27 was oxidized into a 
carboxylic function as indicated by the cross-peak of 
C-27 (δC 179.7) with the protons of methyl group 
CH3-26 at δH 0.90. The proton H-3 had two anti 
coupling constants which indicated a trans-diaxial 
relationship between H-3/H-4. The NMR data of 1 
was in agreement with that reported for trichadenic 
acid B [4]. 
Compound 2 was obtained as white solid, mp 
249-252 
o
C and optically active, [α]D
25
 +5.3
 
(c, 1.5, 
CHCl3). The 1D NMR spectra of 2 were close to 
those of 1, except for the presence of a ketone group 
for 2 instead of the oxymethine group of 1. This data 
suggested that 2 was a derivative of 1 by oxidation 
of hydroxyl group at C-3 into ketone functionality. 
Complete analyses of 2D NMR spectra established 
the structure of 2 as trichadonic acid which was 
previously described [5]. 
Compound 3 was isolated as yellow powder. Its 
1
H-NMR spectrum presented signals of twelve 
aromatic protons, including an A2B2 system [δH 6.71 
(2H, d, J = 8.7 Hz, H-3”’ and 5’”), 7.50 (2H, d, J = 
8.7 Hz, H-2’” and 6’”)], an ABX system [δH 7.08 (J 
= 8.5 Hz, H-5’), 7.82 (J = 2.0, 8.5 Hz, H-6’), 7.97 (J 
= 2.0 Hz, H-2’)], three singlets at δH 6.34 (H-6’’) 
and 6.55 (H-3) and 6.56 (H-3’’), and two doublets 
 126 
VJC, Vol. 53(2e), 2015                                                                                   Constituents from stem barks of… 
with meta-coupling at δH 6.17 (J = 2.0 Hz, H-6) and 
6.39 (J = 2.0 Hz, H-8). Detailed analyses of          
13
C-NMR and DEPT spectra with the aid of HSQC 
spectrum revealed the presence of two carbonyl 
groups at δC 183.8 (C-4) and 184.2 (C-4’’), sixteen 
quaternary carbons and twelve aromatic methines. 
This NMR data suggested 3 was a biflavonoid. This 
was then confirmed by analyses of 2D NMR spectra, 
especially by HMBC spectrum. The linkage between 
the two flavonone units was established by the 
HMBC cross-peaks of C-8’’ with H-6’’ (δH 6.34) 
and H-2’ (δH 7.97) of the ABX system. Intensive 
analysis of the 2D-NMR spectra defined the 
structure of 3 as amentoflavone which was 
previously reported [6]. 
 
 
Figure 1: Compounds isolated from A. poilanei 
  
Compound 4 was determined as β-sitosterol by 
comparison of its NMR data with the reported 
values [7], as well as by TLC analysis comparing 
with the authentic sample which was available in our 
laboratory.          
Acknowledgements. The authors thank Dr. Nguyen 
Quoc Binh, MSc Dao Dinh Cuong for plant 
collection and botanical determination. The Vietnam 
Academy of Science and Technology (VAST) and the 
National Center for Scientific Research, France 
(CNRS) are gratefully acknowledged for support of 
the LIA project NATPROCHEMLAB.
 
 
REFERENCES 
 
1. H. H. Pham. An illustrated flora of Vietnam, Vol. 2, 
Youth Publishing House (2000). 
2. A. B. Alimboyoguen, K. A. De Castro-Cruz, C. C. 
Shen, C. Y. Ragasa. Chemical constituents of 
Anacolosa frutescens, Research Journal of 
Pharmaceutical, Biological and Chemical Sciences, 
5(5), 1189-1191 (2014). 
3. M. Bourjot, P. Leyssen, C. Eydoux, J. C. Guillemot, 
B. Canard, P. Rasoanaivo, F. Guéritte, M. Litaudon. 
Chemical constituents of Anacolosa pervilleana and 
their antiviral activities, Fitoterapia, 83, 1076-1080 
(2012). 
4. R. Tanaka, S. Matsunaga, T. Ishida. Revised structure 
of trichadenic acid B, a stem bark constituent of 
Phyllanthus flexuosus, Tetrahedron Letter, 29(37),  
4751-4754 (1988). 
5. R. M. Giner-Pons, A. I. Gray, C. Lavaud, G. Massiot, 
S. Gibbons, P. G. Waterman. 30-Norfriedelane 
triterpenes from the stem bark of calocoba glauca, 
Phytochemistry, 31(3), 223 (1992). 
6. J. R. Hanrahan, M. Chebib, N. L. M. Davucheron, B. 
J. Hall, G. A. R. Johnston. Semisynthetic preparation 
of amentoflavone : A negative modulator at GABAA 
receptors, Bioorganic & Medicinal Chemistry 
Letters, 13, 2281-2284 (2003). 
7. K. Yamaguchi. Spectral Data of Natural Products, 
Elsevier Publishing Company, 1, 452 (1970). 
 
Corresponding author: Van Cuong Pham 
Institute of Marine Biochemistry, Vietnam Academy of Science and Technology  
    18, Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam 
    E-mail: phamvc@imbc.vast.vn. 
 127 
VIETNAM JOURNAL OF CHEMISTRY         VOL. 53(2e) 124-126                                            APRIL 2015 
DOI: 10.15625/0866-7144.2015-2e-029 
  


Constituents from stem barks of Anacolosa poilanei Gagnep. (Olacaceae)

http://vjs.ac.vn/index.php/vjchem/article/download/6735/5900

Constituents from stem barks of Anacolosa poilanei Gagnep. (Olacaceae)

Abstract

Similar works

Full text

Available Versions

Vietnam Academy of Science and Technology: Journals Online