Neolignans and flavonoids from the root bark of Illicium henryi

Article abstract of DOI:10.1016/j.fitote.2010.08.008

Two new neolignans (1 and 2) were isolated from the root bark of Illicium henryi, along with four known neolignans and seven known flavonoids (3-13). Their structures were elucidated on the basis of spectroscopic and chemical methods. The absolute configurations of compounds 1 and 2 were determined by the CD spectrum.

Full text of DOI:10.1016/j.fitote.2010.08.008

Fitoterapia 81 (2010) 1228–1231
Contents lists available at ScienceDirect
Fitoterapia
journal homepage: www.elsevier.com/locate/fitote
Neolignans and flavonoids from the root bark of Illicium henryi
Wen-Juan Xiang , Lei Ma ⁎, Li-Hong Hu a,b,⁎
a
a,
a
School of Pharmacy, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, PR China
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, PR China
b
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 25 June 2010
Accepted in revised form 10 August 2010
Available online 18 August 2010
Two new neolignans (1 and 2) were isolated from the root bark of Illicium henryi, along with
four known neolignans and seven known flavonoids (3–13). Their structures were elucidated
on the basis of spectroscopic and chemical methods. The absolute configurations of compounds
1
and 2 were determined by the CD spectrum.
©
2010 Elsevier B.V. All rights reserved.
Keywords:
Illicium henryi
Illiciaceae
Neolignan
Flavonoid
Analgesic
1
. Introduction
Illicium henryi belongs to Family Illiciaceae, and distributes
[
(
4], and massonianoside B (8) [5] and seven known flavonoids,
2R,3R) taxifolin 3-O-β-D-xyloside (4) [6], (2 S,3 S) taxifolin 3-
O-β-D-xyloside (7) [6], quercetin (9) [7], (2R,3R) catechin (10)
8], (2R,3R) taxifolin 3-O-β-D-glucoside (11) [9], (2 S,3 S)
mainly in the south of China. Most of Illicium species are
considered to be toxic plant. The root bark of I. henryi is locally
taken to dispel wind and cold and relieve pain in traditional
Chinese medicine. Currently, the water-soluble extract from
the root bark of I. henryi has been applied to clinical use as an
analgesic agent by intramuscular injection in China. It has been
reported that three sesquiterpene lactones were isolated from
the fruits of I. henryi, one of them exhibits high toxicity in mice
[
taxifolin 3-O-β-D-glucoside (12) [9], and quercitrin (13) [8],
have been isolated from the root bark of I. henryi. The absolute
configuration of 1 and 2 were deduced in the CD spectra.
2
. Experimental
2
.1. Generals
[1]. However, there are no previous reports concerning the
phytochemistry of the root bark of I. henryi so far. For the
purpose of finding analgesic compounds, we carried out
systematic studies on the chemical constituents of the root
bark of I. henryi. Two new neolignans, together withfourknown
lignans, seco-isolariciresinol-O-α-L-rhamnoside (3) [2], 4-O-
Optical rotations were measured on a Perkin-Elmer 341
polarimeter. IR data were recorded on a Nicolet FIIR 750
spetrophotometer. The CD spectra were recorded in MeOH on
JASCO J-810 spectropolarimeter. 1H, C and 2D NMR data
spectra were taken with a Bruker AMX 400 instrument.
Chemical shifts are recorded in δ (ppm). ESIMS were obtained
on a Bruker Esquire 3000 Plus Spectrometer. HRESIMS were
determined on a Micromass Q-Tif Global mass spectrometer.
GC experiments were run on a GC-MS-QP5050A instrument
13
(
2-hydroxymethylethyl)-dihydroconigeryl alcohol 6-(4′-hy-
droxy-3′-methoxyphenyl)-glucoside (5) [3], icariside E4 (6)
⁎
Corresponding authors. Ma and Hu are to be contacted at School of
(
Shimadzu), using a db-1 column (0.25 mm i.d.×30 m;
column temperature, 200 °C; injection temperature 250 °C;
carrier gas N at flow of 32.2 mL/min; detector, EI-MS). Open
column chromatography (CC) was performed with MCI gel
Pharmacy, East China University of Science and Technology, 130 Meilong
Road, Shanghai 200237, PR China. Tel./fax: +86 21 50272221.
E-mail addresses: malei@ecust.edu.cn (L. Ma),
2
simmhulh@mail.shcnc.ac.cn (L.-H. Hu).
0367-326X/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.fitote.2010.08.008

W.-J. Xiang et al. / Fitoterapia 81 (2010) 1228–1231
1229
CHP 20P (high porous polymer, 75–150 μm, Mitsubishi
chemical Ind.), silica gel (200–300 mesh, Qingdao Marine
Chemical Ltd., China), RP-18 (20–45 μm, Fuji Silysia Chemical
Ltd.,), and sephadex LH-20 (20–100 μm, Pharmacia). TLC was
carried out with GF-254 silica gel plates (Yantai Huiyou Inc.,
China). All solvents used were of chemical grade and
purchased from the Shanghai Chemical plant, Shanghai, PR
China.
passed through a sephdex LH-20 column eluted with MeOH–
O (1:1) to get a sugar fraction. The sugar portion was
detected by TLC with authentic samples.
H
2
2.5. Determination of sugar components
The absolute configuration of xylose was determined
according to a reported procedure [10]. The sugar portion and
authentic sugar sample were derivatized with leucine
reagent, and the leucine derivatives were subjected to
GC (column temperature 200 °C; injection temperature
2
.2. Plant material
The root bark of I. henryi were collected in Kaihua County,
Zhejiang Province, PR China in December 2009 and authen-
ticated by Prof. Lihong Hu. A vocher sample of the plant
2
2
50 °C; carrier gas N at flow rate of 32.2 mL/min; derivatives
D-xylose: 8.23 min ).
20
Compound 1, white, amorphous power; [α]
D
+13 (c 0.11,
(
2009012003) was deposited at the Shanghai Institute of
MeOH); IR (KBr) νmax 3421, 2929, 1602, 1517, 1457, 1274,
Materia Medica, Chinese Academy of Sciences, Shanghai, PR
China.
−
1
1
2
035, 819, and 603 cm ; CD (MeOH) λmax (Δε) 212 (7.58),
1
13
35 (−8.75), and 274 (2.69) nm. H NMR and C NMR data,
see Table 1; ESIMS m/z 501.2 [M+Na] ; HRESIMS m/z
01.1733 [M+Na] (calcd for C24
Compound 2, white, amorphous power; [α]
MeOH); IR (KBr) νmax 3396, 2927, 2879, 1700, 1608, 1511,
+
2
.3. Extraction and isolation
+
5
H30NaO10 , 501.1737).
20
D
0 (c 0.11,
The air-dried root bark of I. henryi (1.0 kg) were extracted
three times with 95% EtOH under reflux for 2 h. The extract
was concentrated under reduced pressure to obtain a crude
2
extract, which was suspended in H O, and then partitioned
with petroleum ether, EtOAc and n-BuOH, respectively. The
EtOAc layer residue (69.0 g) was subjected to silica gel CC
−1
1
(
452, 1263, 1135, and 1031 cm ; CD (MeOH) λmax (Δε) 227
1
13
3.58), 242 (−0.10), and 261 (0.91) nm. H NMR and
C
+
NMR data, see Table 1; ESIMS m/z 443.2 [M+Na] ; HRESIMS
+
8
m/z 443.1686 [M+Na] (calcd for C22H28NaO , 443.1682).
with CHCl
3
–MeOH gradient to give four fractions. Fr. 1
(
1.227 g) was repeatedly applied to silica gel CC and eluted
3. Results and discussion
3
with CHCl –MeOH and further purified by sephadex LH-20
using MeOH to give 1 (15 mg). Fr. 2 (4.415 g) was
fractionated on silica gel CC with CHCl –MeOH to give two
subfractions. Fr. 2.1 was subjected to sephadex LH-20
CC using 1:1 MeOH–H O and further chromatographyed on
RP-18 with 35:65 MeOH–H O to yield 2 (11 mg), 3 (98 mg)
and 4 (190 mg). The separation of Fr. 2.2 by sephadex LH-20
using 1:1 MeOH–H O and RP-18 was eluted with 15:85
MeOH–H O to afford 5 (20 mg) and 6 (13 mg). Fr. 3 (1.321 g)
was separated by sephadex LH-20 column using MeOH, silica
gel CC with a CHCl –MeOH gradient and further purified by
sephadex LH-20 CC eluted with 1:1 MeOH–H O to yield 7
134 mg). Further purification of Fr. 4 (2.074 g) was achieved
Compound 1, obtained as a white amorphous powder,
has a molecular formular of C24 10 on the basis of a
positive-ion HRESIMS at m/z 501.1733 (cald. for C24H O10Na,
30
3
30
H O
2
2
Table 1
1
H (400 MHz) and 13C NMR (100 MHz) data of compounds 1 and 2 (in
2
3
CD OD).
2
Position
1
2
3
δ
C
δ
H
(J in Hz)
δ
C
H
δ (J in Hz)
2
(
1
2
3
4
5
6
7
8
9
130.1
112.9 7.12 d ( 2.1)
149.5
138.6
119.2 7.08 d (1.5)
152.4
2
by sephadex LH-20 using 1:1 MeOH–H O to give 8 (32 mg)
and 9 (221 mg). The crude n-BuOH extract (128.6 g) was
subjected to MCI CC and eluted with MeOH–H O gradient to
give three fractions, Fr. 5, 6 and 7. Fr. 5 (7.180 g) was
148.6
148.8
2
116.7 6.87 d (8.4)
122.0 6.97 dd (8.4, 2.1)
77.7 5.07 d (8.4)
79.1 4.16 m
111.6 7.10 d (8.1)
119.8 6.98 dd (8.1, 1.5)
88.7 5.57 d (5.7)
56.4 3.46 m
successively purified on silica gel with CHCl
and RP-18 eluted with 15:85 MeOH–H
3
–MeOH gradient
O to afford 10
2
69.8 4.04 dd (11.4, 2.1); 3.39 dd
65.6 3.75 m; 3.84 m
(
48 mg). Fr. 6 (14.1 g) was submitted to CC: silica gel,
sephadex LH-20 and RP-18 to yield 11 (40 mg) and 12
15 mg). Fr. 7 (52.5 g) was shown the presence of one main
compound signal by TLC analysis. A little portion of Fr. 7
400 mg) was further purified by sephadex LH-20 and then
RP-18 CC to give 13 (55 mg).
(11.4, 2.1)
1′
2′
137.0
137.3
117.5 6.60 br, s
142.4
146.9
130.0
117.1 6.62 br, s
33.1 2.58 t (6.6)
36.2 1.81 m
62.7 3.58 t (6.6)
83.4 4.24 m
62.4 3.74 m
62.4 3.74 m
(
122.8 6.72 dd (8.1,2.4)
118.1 6.85 d (8.1)
143.3
3
4
5
6
7
8
9
1
2
′
′
′
′
′
′
′
″
″
(
145.5
118.2 6.77 d (1.8)
32.8 2.60 t (7.2)
36.0 1.81 m
62.6 3.58 t (6.6)
106.0 4.08 d (7.8)
75.3 3.23 m
2
.4. Acid hydrolysis of compound 1
Compound 1 (5 mg) was heated at 80 °C in 10% HCl–
dioxane (1:1, 1 mL) for 4 h in water bath. The reaction
mixtures were neutralized with NaHCO and then evaporated
to dryness. The residue was partitioned between CH Cl and
O for three times. The aqueous phase was evaporated and
3″
4″
78.2 3.28 m
71.6 3.46 m
3
5
OCH
″
67.4 3.11 m; 3.79 m
57.1 3.88 s
2
2
3
56.9 3.81 s
H
2

1230
W.-J. Xiang et al. / Fitoterapia 81 (2010) 1228–1231
Fig. 1. The key HMBC correlations of compounds 1–2.
1
5
01.1737). The H NMR spectrum exhibited signals of a 1,3,4-
methoxyl-9-O-β-D-xylopyranosyl-4′:7,5′:8-diepoxyneo-
trisubstituted aromatic ring [δ 7.12 (1H, d, J=2.1 Hz, H-2), δ
lignan-4, 9′-diol.
6
6
.87 (1H, d, J=8.4 Hz, H-5), δ 6.97 (1H, dd, J=8.4, 2.1 Hz, H-
)], a 1′,4′,5′-trisubstituted aromatic ring [δ 6.72 (1H, dd,
Compound 2 gave the molecular formula C22
H
28
O
8
base
+
1
on an [M+Na] ion at 443.1686 in the HRESIMS. The H NMR
spectrum showed signals assigned to a 1,3,4-trisubstituted
aromatic ring [δ 7.10 (1H, d, J=8.1 Hz, H-5), δ 7.08 (1H, d,
J=1.5 Hz, H-2), δ 6.98 (1H, dd, J=8.1, 1.5 Hz, H-6)] and
a 1′,3′,4′,5′-tetrasubstituted aromatic ring [δ 6.62 (1H, br, s,
H-6′), δ 6.60 (1H, br, s, H-2′)], in addition with a methoxyl
J=8.1, 2.4 Hz, H-2′), δ 6.85 (1H, d, J=8.1 Hz, H-3′), δ 6.77
1H, d, J=2.4 Hz, H-6′)], a hydroxypropyl group [δ 2.60 (2H,
t, J=7.2 Hz, H -7′), δ 1.81 (2H, m, CH , H -8′), δ 3.58 (2H, t,
J=6.6 Hz, H -9′)], a moiety of (Ph)CH(O)-CH(O)-CH O- [δ
.07 (1H, d, J=8.4 Hz, H-7), δ 4.16 (1H, m, H-8), δ 4.04
1H, dd, J=11.4, 2.1 Hz, H-9a), δ 3.39 (1H, dd, J=11.4, 2.1 Hz,
H-9b)], and a methoxyl group [δ 3.88 (3H, s)]. The carbon
(
2
2
2
2
2
5
(
group [δ 3.81 (3H, s, OCH
3
)]. The 1H NMR data also were in
good agreement with a 1,2,3-propantriol moiety [δ 4.24 (1H,
m, H-1″), δ 3.74 (4H, m, H-2″ and H-3″)], a propan-3-ol
moiety [δ 2.58 (2H, t, J=6.6 Hz, H-7′), δ 1.81 (2H, m, H-8′),
δ 3.58 (2H, t, J=6.6 Hz, H-9′] and a moiety of [(CH(O)-CH
1
3
signals in the C NMR spectrum of 1 further confirmed the
above units. The 1D and 2D NMR spectra showed the
presence of a xylopyranosyl moiety. The anomeric proton
signal appeared as a doublet at δ 4.08 (1H, J=7.8 Hz) and
permitted assignment of a β configuration of the xylose. On
acid hydrolysis and GC analysis, the sugar part was
determined to be a β-D-xylopyranosyl group. Significant
HMBC correlations were observed between H-6/C-7, H-7/C-6,
2
(Ph)-CH O)] [δ 5.57 (1H, d, J=5.7 Hz, H-7), δ 3.46 (1H, m, H-
8), δ 3.84 (1H, m, H-9a), δ 3.75 (1H, m, H-9b)]. The
assignment of the 1H NMR and C NMR signals of 2 was
based on its HSQC experiment. These units were assigned
by the HMBC correlation from H-7 to C-6, H-6′ to C-7′, H-1″ to
13
3
H-8/C-7, H-8/C-9, and OCH /C-3 (Fig. 1). Thus the skeleton
3
C-4 and OCH to C-3. A trans configuration of 2 was confirmed
1
could be concluded as 3-methoxyl-4′:7,5′:8-diepoxyneo-
lignan-4,9,9′-triol. The β-D-xylopyranosyl group could be
assigned at C-9 from the correlation between H-9 and C-1″ of
xyl. This could be further confirmed by a downfield
methylene signal at δ 69.8 (C-9) in the 13C NMR spectrum.
The relative configuration of H-7 and H-8 was trans from the
by the J7,8 value of 5.7 Hz in the H NMR spectrum. It could be
further determined on the basis of NOESY correlations
between H-8 and H-2, H-6 protons; H-7 and H-9a protons. It
has been reported that the configurations at C-7 and C-8 of the
dihydrobenzofuran skeleton can be clearly distinguished from
the 240–220 nm region. In the CD spectrum, a positive cotton
effect at 227 nm indicated an 8S configuration [4,15,16]. Thus,
the structure of 2 was determined to be 7R,8S-configuration as
shown in Fig. 3. Therefore the structure of 2 was elucidated as
(7R,8S)-3′,9,9′-trihydroxyl-3-methoxyl-4-O-glycerol-7,8-
dihydrobenzofuran-1′-propanolneoligan.
J
7,8 value of 8.4 Hz in the 1H NMR. The absolute stereo-
chemistries of C-7 and C-8 positions were determined by the
CD spectrum (Fig. 2) with the comparison of model
compounds. A negative cotton effect at the peak of 235 nm
for the benzodioxane moiety in the CD spectrum indicated
assignment of 7R,8R absolute configuration for 1 according to
the study of a related benzodioxane system [11–14].
Therefore the structure of 1 was elucidated as (7R,8R)-3-
The other four known neolignans were identified as seco-
isolariciresinol-O-α-L-rhamnoside (3), 4-O-(2-hydroxy-
methylethyl)-dihydro-conigeryl alcohol 6-(4′-hydroxy-3′-
Fig. 2. The CD spectra of compounds 1 and 2.

W.-J. Xiang et al. / Fitoterapia 81 (2010) 1228–1231
1231
Fig. 3. Structures of compounds 1–2.
methoxyphenyl)-glucoside (5), icariside E4 (6), and masso-
References
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