全 文 ： 药学学报 Acta Pharmaceutica Sinica 2011, 46 (2): 179−182 · 179 ·




A new β-naphthalenecarboxylic acid biglycoside from
Chirita longgangensis var. hongyao
WANG Man-yuan1, GONG Mu-xin1, ZHANG Dong2, YANG Lan2*
(1. School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China;
2. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China)
Abstract: To investigate the chemical constituents in the stems of Chirita longgangensis var. hongyao,
methanol extract of the stems was subjected to column chromatography with various chromatographic techniques.
One new β-naphthalenecarboxylic acid biglycoside, 1, 4-dihydroxy-2-naphthalenecarboxylic acid methyl ester-
4-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (1) was isolated, along with two known compounds:
isotaxiresinol 4-O-methyl ether (2) and (R)-7-hydroxy-α-dunnione (3). Compound 2 was first obtained from
Chirita genus and compound 3 was isolated from this plant for the first time. All structures were elucidated on
the basis of spectral and chemical evidence, and the NMR spectroscopic data of compound 2 was published for
the first time.
Key words: Chirita longgangensis var. hongyao; Gesneriaceae; β-naphthalenecarboxylic acid biglycoside
CLC number: R284 Document code: A Article ID: 0513-4870 (2011) 02-0179-04
红药中一个新的 β-萘甲酸双糖苷类化合物
王满元 1, 龚慕辛 1, 张 东 2, 杨 岚 2*
(1. 首都医科大学中医药学院, 北京 100069; 2. 中国中医科学院中药研究所, 北京 100700)

摘要: 为了研究红药 (Chirita longgangensis var. hongyao) 茎的化学成分, 运用多种色谱方法进行分离纯
化, 从其甲醇提取物中分离得到 3 个化合物, 并根据理化性质和波谱数据鉴定其结构分别为 1, 4-dihydroxy-2-
naphthalenecarboxylic acid methyl ester-4-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (1), isotaxiresinol
4-O-methyl ether (2) 和 (R)-7-hydroxy-α-dunnione (3)。其中, 化合物 1 为新的 β-萘甲酸双糖苷化合物, 化合物 2
为首次从该属植物中分离得到, 且首次提供了化合物 2 的核磁波谱数据, 化合物 3 为首次从该植物中分离得到。
关键词: 红药; 苦苣苔科; β-萘甲酸双糖苷

Chirita longgangensis W. T. Wang var. hongyao S. Z.
Huang (Gesneriaceae) is distributed in Guangxi Province,
China. The stems of C. longgangensis var. hongyao
have long been used as a folk medicine in China for
the treatment of arthritis, anemia and fracture[1, 2].
Previous chemical investigation has resulted in five

Received 2010-09-12.
Project supported by the Beijing Municipal Project for Developing
Advanced Human Resources (No.20061D0501800256).
*Corresponding author Tel / Fax: 86-10-64014347,
E-mail: ylan_66@yahoo.com.cn
phenylethanoid glycosides[3], two anthraquinones and
five other compounds from this plant[4]. The current
study was carried out to search for bioactive metabolites
from C. longgangensis var. hongyao, leading to the
isolation of a new compound, β-naphthalenecarboxylic
acid biglycoside, 1, 4-dihydroxy-2-naphthalenecarboxylic
acid methyl ester-4-O-α-L-rhamnopyranosyl-(1→6)-β-
D-glucopyranoside (1) and two known compounds,
isotaxiresinol 4-O-methyl ether (2) and (R)-7-hydroxy-
α-dunnione (3). Compound 2 was first obtained from
Chirita genus and compound 3 was isolated from this
DOI:10.16438/j.0513-4870.2011.02.004
· 180 · 药学学报 Acta Pharmaceutica Sinica 2011, 46 (2): 179−182

plant for the first time. The chemical structures of
compounds 1−3 are shown in Figure 1. In this paper,
we report the isolation and structure elucidation of
these compounds.


Figure 1 The chemical structures of compounds 1, 2 and 3

Results and discussion
Compound 1 was obtained as white amorphous
powder that analyzed for the molecular formula
C24H30O13 by HR-ESI-MS at m/z 525.161 0 [M−H]−.
The IR (KBr) spectrum of 1 showed broad absorption
for multiple hydroxyl groups (3 399 cm−1), an ester
carbonyl (1 695 cm−1), and aromatic rings (1 635 and
1 603 cm−1) functionalities. The 1H NMR spectrum of
1 (Table 1) showed an aromatic singlet at δ 7.38, a
1, 2-disubsititued aromatic ring at δ 8.36 (1H, d, J = 8.0
Hz, H-5), 7.72 (1H, m, H-6), 7.64 (1H, m, H-7), 8.29
(1H, d, J = 8.0 Hz, H-8), an O-methyl group at δ 3.98
(3H, s, OCH3-9), and a phenolic OH proton at δ 11.7
(1H, s, OH-1). Acid hydrolysis of 1 afforded glucose and
rhamnose, which were identified by TLC comparison
with authentic samples. One doublet and a broaden
singlet due to anomeric protons at δ 4.82 (1H, d, J =
7.5 Hz, H-1) and 4.54 (1H, br s, H-1), together with
a methyl doublet at δ 1.07 (3H, d, J = 6.5 Hz, H-6),
as well as partially overlapped signals attributable to
oxymethylenes and oxymethines between at δ 3.14 and
3.90, indicated that there were a β-glucopyranosyl and
an α-rhamnopyranosyl groups. The configuration of
the glucopyranosyl and rhamnopyranosyl was assigned
as β-D- and α-L- on the basis of the coupling constant
of the anomeric proton and of the abundance of the
β-D-glucopyranosyl and α-L-rhamnopyranosyl units in
natural products. Moreover, the 13C NMR data of the
sugar unit are consistent with those in literature[5].
The 13C NMR spectrum of 1 (Table 1) showed carbon
signals corresponding to the above structural units and
one conjugated ester carbonyl at δ 170.5. 2D NMR
experiments were carried out to construct the structure
of 1. Analyses of the 1H-1H COSY and HMQC
spectra of 1 led to unambiguous assignment of proton
and corresponding carbon signals in the NMR spectra
(Table 1). HMBC correlations of H-8 with C-1, C-8a,
C-7, and C-6, H-5 with C-4, C-4a, C-6 and C-7, and
H-3 with C-4, C-4a, C-2, and C-1, in combination with
chemical shifts of these protons and carbons, provided
evidence for a 1, 2, 4-trisubstituted naphthalene moiety.
The downfield chemical shift of phenolic OH proton
at δ 11.7 suggested 1 possessed one intramolecular
hydrogen bond structure skeleton. HMBC correlations
of the carbonyl (C-9) with H-3 and O-methyl protons,
and the phenolic OH proton with C-1, C-2 and C-8a,
clearly located a hydroxyl and a methyl ester at C-1
and C-2, respectively. The signals assigned to the
aglycone moiety were in good agreement with the
published data in the literature[6]. In addition, HMBC
correlation between the anomeric proton of rhamno-
pyranosyl (H-1) and C-6 of the glucopyranosyl
indicated a rhamnopyranosyl (1→6) glucopyranosyl
linkage. Finally, the sugar chain was positioned at C-4
on the basis of HMBC correlations of the anomeric
proton of glucopyranosyl (H-1) with C-4. Therefore,
the structure of 1 was determined as 1, 4-dihydroxy-
2-naphthalenecarboxylic acid methyl ester-4-O-α-L-
rhamnopyranosyl-(1→6)-β-D-glucopyranoside. The key
HMBC and 1H-1H COSY correlations of compound 1
are shown in Figure 2.

Table 1 1H NMR (500 MHz) and 13C NMR (125 MHz) data for
compound 1 (DMSO-d6, J in Hz). aSignal patterns were unclear
due to overlapping
No. δH δC No. δH δC
1 155.2 Glc-1 4.82 (d, 7.5) 102.3
2 104.5 2 3.40a 73.4
3 7.38 (s) 107.7 3 3.31a 76.2
4 145.3 4 3.14a 70.0
4a 129.9 5 3.48a 75.7
5 8.36 (d, 8.0) 122.4 6 3.42, 3.88 (d, 7.5) 66.6
6 7.72 (m) 129.4 Rha-1 4.54 (br s) 100.6
7 7.64 (m) 126.7 2 3.60 (m) 70.3
8 8.29 (d, 8.0) 123.1 3 3.43a 70.7
8a 124.6 4 3.16a 72.0
9 170.5 5 3.40a 68.2
9-OCH3 3.98 (s) 52.8 6 1.07 (d, 6.5) 17.8
1-OH 11.7 (s)
WANG Man-yuan, et al: A new β-naphthalenecarboxylic acid biglycoside from Chirita longgangensis var. hongyao · 181 ·


Figure 2 The key HMBC and 1H-1H COSY correlations of
compound 1

Compound 2 was firstly reported as a new lignan
by Erdtman H. and Tsuno K. in 1969[7]. No NMR
spectroscopic data for this compound had been reported,
although some data of its structure-similar compounds
have been described in the later literature[8−10]. Therefore,
the NMR data of 2 assigned by interpretation of its 2D
NMR spectra are included in this report.

Experimental
1 Generals
Optical rotations were measured on a JASCO
P-1020 polarimeter. UV and IR spectra were recorded
on Shimadzu UV-2500PC and Shimadzu IR Prestige-21
spectrophotometers, respectively. 1H and 13C NMR
spectra were obtained on a Bruker AM-500 spectrometer.
Proton detected heteronuclear correlations were measured
using HMQC and HMBC. HR-ESI-MS analysis was
carried out on a Bruker microTOF-Q instrument.
Column chromatography was performed using silica gel
(60−120 and 300−400 mesh); TLC: precoated silica gel
plates 60 GF254 or RP-C18 F254 plates with 0.5 or 1 mm
film thickness (Merck). Spots were visualized under
UV light or by spraying with H2SO4-EtOH or anisalde-
hyde-H2SO4 followed by heating.
2 Plant material
The stems of C. longganesis var. hongyao were
collected in Tiandeng County, Guangxi Province, China,
in 2005, and identified by Prof. Bin Dai, Guangxi
Institute of National Medical Research, China, where a
voucher specimen was deposited.
3 Extraction and isolation
The air-dried stems of C. longganesis var. hongyao
(10 kg) were powdered and consecutively extracted with
MeOH at room temperature. The combined extracts
were concentrated in vacuum to yield a dark red residue
that was suspended in water and then partitioned
successively with EtOAc and n-BuOH. The n-BuOH
extract (200 g) was applied to a Diaion D101 macroporous
adsorbent resin column. Successive elution of the
column with 20% EtOH, 50% EtOH and 95% EtOH
yielded three corresponding fractions after removing
solvents. The fraction eluted with 50% EtOH (42.7 g)
was chromatographed over silica gel, eluting with a
gradient of increasing MeOH (0−100%) in EtOAc, to
give six fractions (1−6). Fraction 2 (6.3 g) was firstly
separated after Sephadex LH-20 CC eluting with a step
gradient from 10% to 50% MeOH in H2O and then
purified by ODS CC eluting with gradient mixtures of
MeOH-H2O [from MeOH-H2O (1∶1, v/v) to MeOH-
H2O (85∶15, v/v)] to yield compound 1 (13.5 mg).
The EtOAc extract (80.3 g) was subjected to CC on
silica gel, and eluted with a gradient of increasing
MeOH (0−50%) in CHCl3, to afford eight fractions
(I−VIII) based on TLC analysis. Fraction VII (11 g)
was further purified by silica gel CC (300 g), eluting
with a gradient of increasing MeOH (20%−40%) in
CHCl3, to give compound 2 (16.4 mg). Compound 3
(23.2 mg) was isolated from fraction VI (19.7 g) by
repeated CC over silica gel using a gradient of increasing
EtOAc (5%−50%) in petroleum ether as eluting solvent.
4 Structure identification
Compound 1 white amorphous powder (CH3OH),
mp 143−145 ℃ and [α]D21 −130 (c 0.1, CH3OH). UV
λmax (CH3OH) nm: 215 (sh), 255, 351.5. IR bands
(KBr) cm−1: 3 399 (br), 2 945, 1 695, 1 635, 1 603,
1 098, 1 067, 1 053, 976. Negative HR-ESI-MS m/z:
525.161 0 [M−H]− (calcd. for C24H29O13 525.160 3).
1H NMR (DMSO-d6, 500 MHz) and 13C NMR (DMSO-
d6, 125 MHz) data were shown in Table 1.
Compound 2 white amorphous powder (CH3OH),
mp 172−173 ℃. UV λmax (MeOH) nm: 282.5. IR
bands (KBr) cm−1: 3 406 (br), 2 361, 1 609, 1 514, 1 445,
1 275, 1 121, 1 028. EI-MS (70 eV): m/z (rel. int. %)
360 [M]+ (83), 311 [M−H2O−OCH3]+ (100); negative
HR-ESI-MS m/z: 359.146 2 [M−H]− (calcd. for C20H23O6
359.148 9). 1H NMR (500 MHz, CD3OD) δ: 6.73 (1H,
d, J = 8.0 Hz, H-5), 6.67 (1H, d, J = 2.0 Hz, H-2), 6.65
(1H, s, H-2), 6.61 (1H, dd, J = 8.0, 2.0 Hz, H-6), 6.18
(1H, s, H-5), 3.79 (3H, s, OCH3-4), 3.78 (1H, m, H-7),
3.76 (3H, s, OCH3-3), 3.68 (1H, m, H-9), 3.66 (1H, m,
H-9), 3.64 (1H, m, H-9), 3.39 (1H, dd, J = 11.0, 4.0 Hz,
H-9), 2.76 (2H, d, J = 7.5 Hz, H-7), 1.99 (1H, m, H-8),
1.76 (1H, m, H-8). 13C NMR (125 MHz, CD3OD) δ:
129.1 (C-1), 112.5 (C-2), 147.2 (C-3), 145.3 (C-4),
117.4 (C-5), 134.2 (C-6), 33.6 (C-7), 40.1 (C-8), 66.0
(C-9), 138.6 (C-1), 113.9 (C-2), 149.0 (C-3), 146.0
· 182 · 药学学报 Acta Pharmaceutica Sinica 2011, 46 (2): 179−182

(C-4), 116.0 (C-5), 123.2 (C-6), 48.1 (C-7), 48.7
(C-8), 62.3 (C-9), 56.4 (OCH3-3), 56.5 (OCH3-4). The
1H and 13C NMR data assigned by interpretation of
its 2D NMR spectra are in good accordance with its
structure-similar compounds in the literature[8−10], so
compound 2 was identified as isotaxiresinol 4-O-methyl
ether.
Compound 3 red needle crystals (CHCl3), mp
133−134 ℃. EI-MS (70 eV): m/z (rel. int. %) 258 [M]+
(50), 243 [M−CH3]+ (100); HR-EI-MS m/z: 258.088 8
[M]+ (calcd. for C15H14O4 258.089 2). 1H NMR (300
MHz, CDCl3) δ: 7.94 (1H, d, J = 8.5 Hz, H-5), 7.48 (1H,
d, J = 2.5 Hz, H-8), 7.12 (1H, dd, J = 8.5, 2.5 Hz, H-6),
4.55 (1H, q, J = 6.5 Hz, H-2), 1.46 (3H, s, H-11), 1.41
(3H, d, J = 6.5 Hz, H-10), 1.26 (3H, s, H-12). 13C NMR
(75 MHz, CDCl3) δ: 91.6 (C-2), 45.2 (C-3), 131.0
(C-3a), 182.1 (C-4), 126.8 (C-4a), 128.6 (C-5), 120.6
(C-6), 160.1 (C-7), 112.7 (C-8), 133.5 (C-8a), 178.6
(C-9), 158.5 (C-9a), 14.2 (C-10), 25.8 (C-11), 20.6
(C-12). The 1H and 13C NMR data are consistent with
those in literature[11], and then compound 3 was
deduced as (R)-7-hydroxy-α-dunnione.
5 Acid hydrolysis of compound 1: determination of
the sugar
A solution of compound 1 (2 mg) was heated with
2 mol·L−1 HCl (2 mL) in a sealed tube at 100 ℃ for 4 h.
The reaction mixture was extracted with ethyl acetate.
After evaporating off the organic layer, the aqueous
phase was neutralized with NaHCO3 and lyophilized.
The lyophilized residue was dissolved in pyridine (0.2
mL), and co-eluted (TLC) with the authentic samples
developed with EtOAc-n-BuOH-H2O (20∶70∶10, v/v).
The plates were sprayed with naphthoresorcinol reagent
by heating at 100 ℃. Glucose and rhamnose were
identified.
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