Accepted Manuscript

Two novel diterpenes from the roots of Phyllanthus acidus (L.) Skeel

Thuc-Huy Duong, Xuan-Hao Bui, Pierre Le Pogam, Huu-Hung Nguyen, Thi-Thuan

Tran, Thi-Anh-Tuyet Nguyen, Warinthorn Chavasiri, Joël Boustie, Kim-Phi-Phung
Nguyen

PII: S0040-4020(17)30745-7
DOI: 10.1016/j.tet.2017.07.021
Reference: TET 28853

To appear in: Tetrahedron

Received Date: 17 March 2017

Revised Date: 18 June 2017
Accepted Date: 11 July 2017

Please cite this article as: Duong T-H, Bui X-H, Le Pogam P, Nguyen H-H, Tran T-T, Nguyen T-A-T,
Chavasiri W, Boustie Joë, Nguyen K-P-P, Two novel diterpenes from the roots of Phyllanthus acidus (L.)
Skeel, Tetrahedron (2017), doi: 10.1016/j.tet.2017.07.021.

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Two novel diterpenes from the roots of Leave this area blank for abstract info.
Phyllanthus acidus (L.) Skeel

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Thuc-Huy Duonga, Xuan-Hao Buia, Pierre Le Pogamb, Huu-Hung Nguyenc, Thi-Thuan Trana, Thi-Anh-

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Tuyet Nguyena, Warinthorn Chavasirid, Joël Boustie e, and Kim-Phi-Phung Nguyenf*
a
Department of Chemistry, Ho Chi Minh City University of Pedagogy, 280 An Duong Vuong Str., District
5, Ho Chi Minh City 748342, Vietnam
b

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Institute of Electronics and Telecommunications of Rennes (IETR), UMR CNRS 6164, University of
Rennes 1, 263 Av. du Général Leclerc, Rennes Cedex 35042, France
c
Faculty of Hi-Tech Agriculture and Biotechnology, Nguyen Tat Thanh University, 300A Nguyen Tat
Thanh Str., Dist. 4, Ho Chi Minh City 748355, Vietnam

U
d
Natural Products Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn University,
Phayathai Rd., Patumwan, Bangkok 10330, Thailand
e
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Institute of Chemistry of Rennes, ISCR, UMR CNRS 6226, University of Rennes 1, 2 Av. du Pr. Léon
Bernard, Rennes Cedex 35043, France
f
Department of Organic Chemistry, University of Science, National University – Ho Chi Minh City, 227
Nguyen Van Cu Str., Dist. 5, Ho Chi Minh City 748355, Vietnam
M
D
TE
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ACCEPTED MANUSCRIPT

Tetrahedron
journal homepage: www.elsevier.com

Two novel diterpenes from the roots of Phyllanthus acidus (L.) Skeel

PT
Thuc-Huy Duonga, Xuan-Hao Buia, Pierre Le Pogamb, Huu-Hung Nguyenc, Thi-Thuan Trana, Thi-Anh-
Tuyet Nguyena, Warinthorn Chavasirid, Joël Boustie e, and Kim-Phi-Phung Nguyenf*
a

RI
Department of Chemistry, Ho Chi Minh City University of Pedagogy, 280 An Duong Vuong Str., District 5, Ho Chi Minh City 748342, Vietnam
b
Institute of Electronics and Telecommunications of Rennes (IETR), UMR CNRS 6164, University of Rennes 1, 263 Av. du Général Leclerc, Rennes Cedex 35042
France
c
Faculty of Hi-Tech Agriculture and Biotechnology, Nguyen Tat Thanh University, 300A Nguyen Tat Thanh Str., Dist. 4, Ho Chi Minh City 748355, Vietnam
d
Natural Products Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn University, Phayathai Rd., Patumwan, Bangkok 10330, Thailand

SC
e
Institute of Chemistry of Rennes, ISCR, UMR CNRS 6226, University of Rennes 1, 2 Av. du Pr. Léon Bernard, Rennes Cedex 35043, France
f
Department of Organic Chemistry, University of Science, National University – Ho Chi Minh City, 227 Nguyen Van Cu Str., Dist. 5, Ho Chi Minh City 748355,
Vietnam

ARTICLE INFO ABSTRACT

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AN
Article history: Two novel diterpenes, phyllanes A and B, were isolated from the roots of Phyllanthus acidus,
Received along with the cleistanthane diterpene, spruceanol. Their chemical structures were
Received in revised form unambiguously elucidated by extensive 1D and 2D NMR analyses and high resolution mass
M

Accepted spectroscopic data, as well as by comparison with literature data. While phyllanes A and B are
Available online respectively reminiscent of amphilectane and serrulatane diterpenoids, their highly unusual
substitution patterns and their co-occurrence with spruceanol led us to assume that they might
correspond to an unprecedented type of re-arranged cleistanthane-diterpenoid precursors,
resulting in final products that display a unique scaffold among terrestrial diterpenoids.
D

Keywords:
Accordingly, a possible biosynthetic route to the two new compounds from the readily
Phyllanthus acidus,
accessible spruceanol is proposed herein. These two compounds were evaluated for their
diterpene,
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cytotoxic activities against two cancer cell lines. Only phyllane B exerted a moderate activity
spruceanol,
against K562 and HepG2 cell line with IC50 values of 28.90 and 45.23 µg/ mL, respectively.
biosynthetic considerations,
cytotoxicity 2009 Elsevier Ltd. All rights reserved.
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1.Introduction sesquiterpene glycosides displaying potent anti-hepatitis B virus

activities.14,15 Although a score of different diterpenoids were
The genus Phyllanthus, consisting of ca. 750 species that fall reported from Phyllanthus species so far,16,17 no diterpenoids
into 11 subgenuses, are widely distributed throughout tropical have been reported from P. acidus.
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and subtropical regions in the world. Some species of this genus

have long been used as herbal drugs in China, India, Brazil and Our phytochemical investigation of P. acidus led to the
Southeast Asia.1 Phyllanthus acidus is traditionally used in
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isolation of two highly unusual diterpenoids, named phyllane A

Vietnam as well as some other countries. Biological assays and B (1− −2), for which a putative biosynthetic scheme is
applied on P. acidus substances or extracts indicated a wide array proposed herein.
of bioactivities including antibacterial,2 antifungal,3
antinematodal, anti-inflammatory and analgesic activities,5 as
4

well as beneficial effects on cystic fibrosis,6 bleomycin-induced

pulmonary fibrosis,7 and obesity and/or cardiovascular diseases.
8,9
Hepatoprotective properties against acetaminophen or
thioacetamide induced hepatotoxicity were also evidenced. 10
Previous phytochemical studies on apolar extracts of
Phyllanthus acidus reported the presence of lupane-type
triterpenes, cyclopropane triterpenes (e.g phyllanthol, β-amyrin)
and phytosterols11–13 while those on polar extracts identified −3
Figure 1. Chemical structures of 1−
highly-oxygenated norbisabolane sesquiterpenes and

∗ Corresponding author. Tel: +84-83852270; fax: +84-838350096; e-mail: kimphiphung@yahoo.fr

 2 Tetrahedron
ACCEPTED MANUSCRIPT
vicinal coupling constant of this methylene group proved that it
2. Results and discussion
Three successive extractions with solvents of increasing
polarities were performed on the crude ethanol extract of the
roots of P. acidus. Further purification and isolation of
compounds were carried out using silica gel chromatography as
−2)
described in the experimental section. Two new products (1−
were isolated and elucidated along with the formerly known,
spruceanol (3).18
Compound 1 was obtained as a white amorphous powder. Its
molecular formula was deduced to be C20H24O3 by HRESIMS,
resulting in nine indices of hydrogen deficiency. 1H NMR and
HSQC spectra of 1 showed one phenolic hydroxy group (δH
8.50), two aromatic ortho-coupled protons (δH 7.74, d, J = 8.5 Hz
and 7.08, d, J = 8.5 Hz), one vinyl moiety (δH 7.04, 1H, dd, J =
17.5, 11.5 Hz; 5.74, 1H, dd, J = 11.5, 2.5 Hz, and 5.33, 1H, dd, J
= 17.5, 2.5 Hz), one diastereotopic sp3 methylene (δH 3.25, 1H,
dd, J = 18.0, 3.5 Hz and δH 3.15, 1H, dd, J = 18.0, 2.5 Hz), one
oxygenated methine (δH 4.38, 1H, m),19 one isopropyl group (δH
1.85, 1H, sept, J = 7.0 Hz; 1.05, 3H, d, J = 7.0 Hz; and 0.96, 3H,
d, J = 7.0 Hz), and two methyl moieties (δH 2.34 and 2.30, each
3H, s) (Table 1). The 13C NMR spectrum, in accordance with
HSQC spectrum, confirmed the presence of twenty carbons
comprising two aromatic methine carbons (δC 126.2 and 124.0),
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one vinyl group (δC 135.9 and 121.2), one isopropyl group (δC
36.5, 17.7, and 16.6), one hydroxylated methine (δC 69.8), one
sp3 methylene (δC 33.4), two methyls (δC 19.7 and 14.0), and nine
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aromatic quaternary carbons in the range of 110–160 ppm. These
NMR and MS data were consistent with a diterpenoid skeleton.
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Figure 2. Key HMBC and NOESY correlations of 1
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The HMBC spectrum of 1 showed the correlations of the first
methyl group (δH 2.30, H3-15) to signals at δC 156.0 (C-12),
124.8 (C-13), and 136.1 (C-14) and of the olefinic proton (δH
C
7.04, H-16) to these two latters, demonstrating the C-12–C-14
connectivity. Likewise, the olefinic proton revealed a HMBC
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correlation to a signal at δC 126.0 (C-8). These findings led us to
define the C-ring as shown in Fig. 2. HMBC cross-peaks
identified a second spin system (the so-called B-ring) comprising
the correlations of the two ortho-coupled protons [δH 7.74, d, 8.5
Hz (H-6) and 7.08, d, 8.5 Hz (H-7)] and H3-20 (δH 2.34) to the
signal at δC 133.4 (C-5), of H-7 to C-8 (δC 126.0), C-9 (δC 129.6),
and C-14 (δC 136.1), and of H-6 to C-8 and C-20 (δC 19.7).
Altogether, these spectroscopic features established the structure
of the B ring as established in Figure 2. The key HMBC of H-16
to C-8 indicated the fused linkage between C- and B- rings at C-
8–C-9. Strong NOESY correlations of H-7 and H-16 and of H2-
17 and H3-15 were in line with this proposed structure.
In the A-ring, the diastereotopic methylene protons H2-1 (δH
3.25, dd, 18.0, 3.5 Hz and δH 3.15, dd, 18.0, 2.5 Hz) showed
HMBC cross-peaks to C-5, C-9, and C-10 (δC 126.6). The
was adjacent to the oxygenated methine CH-2 (δH 4.38, δC 69.8).
HMBC cross-peaks of H-4 (δH 1.85), H3-18/H3-19 (δH 1.05/0.96)
to C-3 (δC 82.5) and C-2 and of H-2 and H2-1 to C-3 indicated the
attachment of the isopropyl moiety at C-3. The A/B junction
could be further determined thanks to HMBC cross-peaks
between H-2 and H-6 to C-10 as well as between H-1 and C-9.
This connectivity was further confirmed by NOE cross-peaks
between H2-1 and H3-20. All aforementioned chemical features
account for eight degrees of unsaturation. The HMBC correlation
of H-2 to the quaternary carbon at δC 114.7 (C-9) established the
connectivity of the two rings A and C at C-11, standing for the
last degree of unsaturation. The overall structure of 1 could be
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established as 1-isopropyl-4,7,8-trimethyl-2,3-dihydro-1H-
phenalene-1,2,9-triol and named phyllane A (shown in Fig. 1).
The NOESY spectrum conveyed some information for the
relative stereochemistry of 1. Key NOE cross–peaks between H–
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2 and H-1a, H-4, H3-18, H3-19 indicated the syn orientation of H-
2 and the isopropyl group. Moreover, strong NOESY correlation
between H-1a (δH 3.25) and H3-18 showed that proton H-1a and
the isopropyl group were also synfacial. This led us to distinguish
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the 1H chemical shifts of two protons H-1a (δH 3.25, dd, J = 18.0,
3.5 Hz) and H-1b (δH 3.15, dd, J = 18.0, 2.5 Hz) of the
diastereotopic methylene H2-1. The small value of the vicinal
couplings of H2-2 diastereotopic protons are indicative of gauche
coupling constants with similar magnitudes being reported on
substituted cyclohexane rings throughout literature.19,20
Therefore, such values are diagnostic of trans pseudoequatorial-
pseudoequatorial and cis pseudoequatorial-axial configurations.
This leads to unambiguously define the position of the proton H-
2 as pseudoequatorial, thus ascribing the synfacial isopropyl
moiety to an axial position. Even though an axial configuration
does not appear to be thermodynamically favored for such a
bulky substituent, isobutenyl or isopropyl groups were reported
to occur in pseudo-axial position in related scaffolds.21
Compound 2 was obtained as a light-yellow amorphous
powder. Its 13C NMR and HRESIMS data established a
molecular formula of C20H24O2. The comparison of NMR spectra
of 2 and 1 showed that the constitutions of the A and B rings of 2
were highly reminiscent to those of 1 (Table 1, Fig. 3). As to this
bicyclic core, the signal patterns only differed for one position.
1
H NMR spectrum of 2 revealed the presence of an additional
aromatic proton at δH 7.28 (1H, s, H-11) whose carbon resonated
at δC 104.8 (C-11), replacing the quaternary carbon C-11 of 1.
This assumption was further strengthened by HMBC cross-peaks
between H-11 to C-12 (δC 155.2), C-8 (δC 133.3), and C-9 (δC
127.0) while 12-OH (δH 8.65) correlated to C-11. These
spectroscopic features indicated that 2 was a 3,11-seco derivative
of 1. A further difference was the disappearance of the chiral
methine CH-2 in 1 to give rise to a methylene signal in 2. This
modification in the 1H NMR spectrum of 1 was accompanied by
the conversion of the diastereotopic methylene H2-1 pattern of
peaks to give rise to a regular methylene signal pattern, further
suggesting the suppression of the vicinal chiral center. HMBC
cross-peaks of H2-1 (δH 3.16, m) and H2-2 (δH 2.73, m) to C-10
(δC 132.8) supported the position of the side-chain at C-10.
HMBC correlations of H-4 (δH 2.70, 1H, sept), H3-18/H3-19 (δH
1.06, 6H, d), H2-2, H2-1 to the carbon at δC 213.7 (C-3) indicated
the position of the ketone group at C-3. Altogether, these
identified 2 as 1-(7-hydroxy-2,6-dimethyl-5-vinylnaphthalen-1-
yl)-4-methylpentan-3-one (shown in Fig. 1).
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Table 1 ACCEPTED MANUSCRIPT
−2 (Acetone-d6)
NMR spectral data of compounds 1−

Position 1a 2b

δC δH, m J (Hz) δC δH, m J (Hz)

1 33.4 3.25 dd, 18.0, 3.5 24.0 3.16 m
3.15 dd, 18.0, 2.5
2 69.8 4.38 m 41.7 2.73 m
3 82.5 213.7
4 36.5 1.85 sept, 7.0 40.7 2.70 sept, 7.0

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5 133.4 133.0
6 126.2 7.08 d, 8.5 126.8 7.08 d, 8.5
7 124.0 7.74 d, 8.5 124.3 7.74 d, 8.5
8 126.0 133.3
9 129.6 127.0

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10 126.6 132.8
11 114.7 104.8 7.28 s
12 156.0 155.2
13 124.8 123.9

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14 136.1 137.9
15 14.0 2.30 s 13.7 2.29 s
16 135.9 7.04 dd, 17.5, 11.5 135.8 7.05 dd, 17.5, 11.5
17 121.2 5.74 dd, 11.5, 2.5 121.3 5.76 dd, 11.5, 2.5

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5.33 dd, 17.5, 2.5 5.32 dd, 17.5, 2.5
18 16.6 1.05 d, 7.0 18.5 1.06 d, 7.0
19 17.7 0.96 d, 7.0 18.5 1.06 d, 7.0
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20 19.7 2.34 s 20.0 2.41 s

2-OH
3-OH
12-OH 10.57 s 8.65 s
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ring aromatization might then afford the corresponding seco-

derivative, as previously hypothesized for 2. This structure might
then be susceptible to an intramolecular Friedel-Crafts alkylation
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via an electrophilic attack at C-11 to generate a C3-C11 bond, as

described elsewhere on closely related metabolites.25 NOESY
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correlations then enable to ascribe the second OH group to the

same face of the cyclohexane ring of 1 even though no
spectroscopic evidence rules out the possibility that these
moieties shall occur on the other side of this nucleus.
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Figure 3. Key HMBC and NOESY correlations of 2 A score of the rare cleistanthane-diterpene derivatives have
been reported so far.26 Being constantly isolated alongside the
more common pimarane diterpenoids,27-29 it was early surmised
Biosynthetic considerations that a close biogenetic link might exist between these structural
series.30 One should note that serrulatane diterpenoids,
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A putative biosynthesis for compounds 1 and 2 is shown in

Scheme 1. Spruceanol (3) may serve as a precursor, which is structurally related to 2, were proved to be biosynthetic
intermediates en route to amphilectane diterpenoids,31 that
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modified by a sequence of oxidation, degradation, aromatization,

1,2-alkyl rearrangement, and intramolecular Friedel-Crafts
alkylation. Spruceanol (3) is susceptible to oxidation to form
sonderianol, which bears a keto group at position C-3.22 This
intermediate could enable the degradation of C-4-C-5 linkage to
form its seco-derivative. The driving force of this bond cleavage
might be the protonation of the carbonyl group of C-3 to afford a
C-5 carbonium ion. The reactivity of this putative intermediate
might then account for the 1,2-methyl shift from C-10 to C-5 that
would lead to 2 after subsequent B-ring aromatization.
Sonderianol might be oxidized at C-2 as a first step to reach
compound 1. This biosynthetic step is reminiscent of the
conversion of spruceanol to cleisthanthol,23 a cleistanthane-
diterpenoid with the process previously described by Maslovkaya
and co-workers.24 The next steps of the biosynthesis would be the
degradation of C-4-C-5 bond. Subsequent 1,2-methyl shift and B-
display the same tricyclic core than 1, using various radiolabeling
experiments.32 Mechanisms of cyclization involving related seco-
derivatives were also evoked to explain the existence of abietane-
type tricyclic diterpenoids such as candidissiol33 and
salviaprione.34 The involvement of 4,5-seco-spruceanol
derivative is further suggested by the isolation of 2 which
displays such a seco-C ring scaffold, even though it contains
slightly different substituents on its side chain.23 Most
interestingly, both the scaffolds of 1 and 2 are unprecedented
among terrestrial diterpenoids.35,36 The minute amounts of
molecules obtained despite the large quantity of starting plant
material might account for the fact that these structures were not
isolated so far.
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Scheme 1: Putative biosynthetic pathway for the formation of compounds 1 and 2 from spruceanol (3).
[(i): oxidation, (ii): protonation following by C-4-C-5 degradation and 1,2-methyl shift, (iii): B-ring aromatization, (iv): intramolecular Friedel-
Crafts alkylation].
In this study, compounds 1 and 2 were tested for their
cytotoxicity against K562 (chronic myelogenous leukemia) and
HepG2 (liver hepatocellular carcinoma) cell lines (Figure S3,
Tables S1, S2). Only compound 2 exhibited moderate activity
against both K562 and HepG2 cell lines with respective IC50
values of 28.90 and 45.23 µg/ mL. Compound 1 failed to reveal
any cytotoxic activity.
3. Conclusion
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In conclusion, two novel diterpenes, phyllanes A and B, were
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isolated from the roots of Phyllanthus acidus. A putative
biosynthetic pathway for their formation was proposed. To the
best of our knowledge, the diterpenoid scaffolds of 1 and 2 are
the first reported from a terrestrial source.
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4. Experimental
4.1 General experimental procedures
D
NMR spectra were measured on Bruker Avance III (500 MHz
for 1H NMR and 125 MHz for 13C NMR) spectrometer. Proton
chemical shifts were referenced to the solvent residual signal of
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CD3COCD3 at δH 2.05. The 13C NMR spectra were referenced to
the central peak of CD3COCD3 at δC 29.4. HR–ESI–MS were
recorded on a Bruker microTOF Q-II. TLC analyses were carried
out on pre-coated silica gel 60 F254 or silica gel 60 RP–18 F254S
(Merck) and spots were visualized by spraying with 10% H2SO4
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ethanolic solution followed by heating. Gravity column
chromatography (CC) was performed using silica gel 60 (0.040–
0.063 mm, Himedia).
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4.2 Plant material
The roots of Phyllanthus acidus (Euphorbiaceae) were
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collected from Hong Son village, Ham Thuan Bac, in Binh
Thuan province in July 2014. The botanical sample was
authenticated by Dr. Pham Van Ngot, Department of Botany,
Faculty of Biology, Ho Chi Minh University of Pedagogy. A
voucher specimen (No UP-B01) was deposited in the herbarium
of the Department of Organic Chemistry, Faculty of Chemistry,
Ho Chi Minh University of Pedagogy.
4.3 Extraction and isolation
Roots of Phyllanthus acidus were dried and milled to obtain
35 kg of powder. The powder was extracted with EtOH (3 x 20
L) at 70 oC. The filtrated solution was evaporated to dryness
under reduced pressure to obtain a crude extract (2.0 kg). This
crude was applied to normal phase silica gel CC, and eluted with
a solvent system of n-hexane: EtOAc (stepwise, 10:0 to 0:10) to
afford fractions, H (41.1 g), HA (10.2 g), EA1 (114.2 g), and
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EA2 (170.6 g). Further elution with MeOH gave the methanol
extract M (640.4 g).
Fraction HA (10.2 g) was applied to Sephadex LH-20 CC,
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eluted with MeOH to afford four sub-fractions HA1.1 (0.7 g),
HA1.2 (1.1 g), HA1.3 (1.7 g) and HA1.4 (6.1 g). Sub-fraction
HA1.1 was fractionated with n-hexane: CHCl3: EtOAc: acetone:
AcOH (5:1:2:2:0.02) to afford three fractions, HA1.1.1 (100.8
mg), HA1.1.2 (51.2 g) and HA1.1.3 (423.9 g). Fraction HA1.1.1
was further purified by preparative TLC using n-hexane: CHCl3:
EtOAc: acetone: AcOH (5:1:2:2:0.02) as eluent to afford three
compounds, 1 (3.8 mg), 2 (4.1 mg), and 3 (21.0 mg).
4.3.1. Phyllane A (1). White amorphous powder; HR-ESI-MS
m/z 335.1649 [M+Na]+ (calcd for C20H24O3Na, 335.16177). For
1
H and 13C NMR (acetone-d6) spectroscopic data, see Table 1.
4.3.2. Phyllane B (2). Light-yellow amorphous powder; HR-ESI-
MS m/z 319.1683 [M+Na]+ (calcd for C20H24O2Na, 319.16685).
For 1H and 13C NMR (acetone-d6) spectroscopic data, see Table
1.
4.4 Biological assays
K562 cells and HepG2 cells were cultured in RPMI 1640
medium or in DMEM medium, respectively, supplemented with
10% fetal bovine serum (FBS), 100 IU/mL penicillin, 100 µg/mL
streptomycin and maintained at 37 °C and 5% CO2 with 95%
humidity. Viable cells were counted and inoculated in 96-well
plate with density of 104 cells/100 µL/well for HepG2 and 105
cells/100 µL/well for K562. After 24 hours, the cells were treated
with the compounds and doxorubicin (positive control) diluted in
culture media at 100, 50, 25, 12.5, 6.25, 3.125 and 0 µg/mL
concentration containing 1, 0.5, 0.25, 0.125, 0.0625, 0.03125, 0%
dimethyl sulfoxide (DMSO), respectively. DMSO in culture
media was used as negative control. In addition, culture medium
without cells was used as blank. All experiments were done in
triplicate. The plates were incubated in 5% CO2, 95% humidity at
37 °C for 72 h. 10 µL of 3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide (MTT, 5 mg/ml stock solution) was
added into each well and incubated in 37 °C in 5% CO2 for 3.5
hours. 70 µL of Detergent Reagent (10% SDS) was added into
each well and the plate was maintained in 37 °C for 16 h. The
optical density of each well was read by using a scanning
multiwall spectrophotometer (Sunrise) at wavelength of 595 nm.
Cell survival was measured as the percentage absorbance
compared to the negative control (DMSO-treated cells).37,38
Acknowledgments

ACCEPTED
This research was supported by Vietnam Ministry of Science MANUSCRIPT
and Technology grant #B2016-SPS-04. The authors would like to
thank Dr. Pham Van Ngot, Department of Botany, Faculty of
Biology, Ho Chi Minh University of Pedagogy for the botanical
authentication of the studied plant.
T. H. Duong and X. H. Bui contributed equally to the
research.
Supplementary Material
Supplementary data related to this article can be found online.
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