全 文 ：16 Chin J Nat Med Jan. 2010 Vol. 8 No. 1 2010 年 1 月 第 8 卷 第 1 期








Chemical Constituents from Helwingia japonica

XIA Li-Zi1,2, ZHOU Min3, XIAO Yan-Hua1, LI Guo-You1, CHEN Xiao-Zhen1,
ZHANG Guo-Lin1*
1Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041;
2Graduate School of Chinese Academy of Sciences, Beijing 100039;
3Chengdu Di’ao Group, Chengdu 610041, China
[ABSTRACT] AIM: To discover protein tyrosine phosphatase 1B (PTP1B) inhibitory compounds from the 95% etha-
nol extract of the aerial parts of Helwingia japonica (Thunb.) Dietr.. METHODS: Two plant samples (A and B) were
collected respectively from Luding County and Pingwu County, Sichuan Province of China, and chromatography was
used for separation, and spectroscopic methods were employed for the identification of isolated compounds. RESULTS:
Fractions HC, HE, HF, HG of the 95% ethanol extract of sample A were active. From A nine compounds (1-9) were iso-
lated. The AcOEt fraction of the 95% ethanol extract of the aerial parts of sample B showed PTP1B inhibitory activity
with an IC50 of 4.63 μg·mL−1, and five compounds (10-14) were isolated from this fraction. The isolated compounds
were identified as β-sitosterol (1), β-daucosterol (2), lupeol (3), betulin (4), betulinic acid (5), hexadecanoic acid 2,
3-dihydroxypropyl ester (6), cinnamic acid (7), stignast-4-en-6β-3-one (8), stignast-4-en-6α-3-one (9), p-menth-2-en-1β,
4β, 8-triol (10), blumenol A (11), 2′, 3′, 4′, 5′, 6′-pentahydroxychalcone (12), apigenin 7-O-β-D-glucopyranoside (13),
and luteolin 7-O-β-D-glucopyranoside (14). CONCLUSION: Except compounds 13 and 14, all the other compounds
are first isolated from Helwingia japonica (Thunb.) Dietr.. Compound 5 is a known PTP1B inhibitor. It is necessary to
study the active compounds in this plant further when taking the yield of compound 5 into account.
[KEY WORDS] Helwingia japonica (Thunb.) Dietr.; Chemical constituents; Protein tyrosine phosphatase 1B
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2010)01-0016-05
doi: 10.3724/SP. J. 1009.2010.00016
The genus Helmingia comprises 5 species, which are
mainly distributed in eastern Asia[1]. Previous phytoche-
mical studies on Helmingia genus revealed the presence of
secoiridoids, polyphenols, flavonoid glycosides and tri-
terpenoids [2-4]. H. japonica (Thunb.) Dietr. is used as a folk
medicine for fever, traumatic injury and activating collat-
erals[1, 5]. It is also used as a constituent of a Chinese herbal
medicinal plastering agent for treating bone fracture[6].
In July 2005, we collected the aerial parts of Helmin-
gia japonica (Thunb.) Dietr. (Sample A) from Luding
County, Sichuan Province of China. Fractions HC, HE, HF,
HG of the ethnol extract of this plant showed inhibitory
activity on protein tyrosine phosphatase 1B (PTP1B).
PTP1B inactivated the insulin signal transduction cascade
by dephosphorylating phosphotyrosine residues in insulin
signaling molecules to lead to type Ⅱ diabetes[7]. To ob-
tain the active compounds, chemical study on this plant was
carried out and nine compounds were isolated from the
plant. On the basis of spectral evidence or by comparison of

[Received on] 18-May-2009
[*Corresponding author] ZHANG Guo-Lin: Prof., Tel: 86-28-
85229901. E-mail: zhanggl@cib.ac.cn
them with authentic samples, they were identified as
β-sitosterol (1), β-daucosterol (2), lupeol (3), betulin (4),
betulinic acid (5), glycerol monopalmitate (6), cinnamic
acid (7), stignast-4-en-6β-3-one (8), and stig-
nast-4-en-6α-3-one (9). It is reported that betulinic acid (5)
and its derivatives showed PTP1B inhibitory activity [8].
However, in view of the yields of triterpenoids, the
components with PTP1B inhibitory active in this plant need
to be studied further.
Therefore, we collected sample B in August 2006
from Pingwu County, Sichuan Province of China. The
AcOEt fraction of the 95% ethanol extract of sample B
showed PTP1B inhibitory activity with an IC50 of 4.63
μg·mL−1. From this fraction five compounds were obtained
and identified as p-menth-2-en-1β, 4β, 8-triol (10), blu-
menol A (11), 2′, 3′, 4′, 5′, 6′-pentahydroxychalcone (12),
apigenin 7-O-β-D-glucopyranoside (13), and luteolin
7-O-β-D-glucopyranoside (14).
1 Apparatus and Reagents
Melting points were measured on an X-6 melting
point apparatus and were uncorrected. Optical rotations
were measured on a Perkin-Elmer 341 automatic polarime
XIA Li-Zi, et al. /Chinese Journal of Natural Medicines 2010, 8(1): 16−20
2010 年 1 月 第 8 卷 第 1 期 Chin J Nat Med Jan. 2010 Vol. 8 No. 1 17



Fig. 1 Compounds isolated from Helwingia japonica

ter. UV and IR spectra were recorded on a Lambda 35 spec-
trometer and a Perkin Elmer spectrum one FT-IR spec-
trometer (KBr disc), respectively. Mass spectra were carried
out on a Finnigan-LCQDECA mass spectrometer (ESI-MS).
NMR spectra (1H: 600 MHz; 13C: 150 MHz) were recorded
on a Bruker Avance 600 spectrometer with TMS as internal
standard. Silica gel H (200-300 mesh; Qingdao Marine
Chemical Group Co., China), MCI gel (CHP 20P, 75-150
μm, Mitsubishi Chemical Corporation, Japan) and Sepha-
dex LH-20 (Pharmacia Biotech, Sweden) were used for
column chromatography (CC). The plates for thin-layer
chromatography (TLC) were precoated with Silica gel
GF254 (0-40 μm, Qingdao Marine Chemical Group Co.,
China) and activated at 110°C for 2 hours. Spots on TLC
plates were visualized by spraying 10% EtOH solution of
phosphomolybdic acid followed by heating．All the re-
agents were purchased from commercial company and dis-
tilled before use.
2 Plant Material
Samples A and B were collected in July 2005 from
Luding County and in August 2006 from Pingwu County,
Sichuan Province of China, respectively and identified by
Prof. Fu Fa-Ding at Chengdu Institute of Biology, Chinese
Academy of Sciences (CAS). The voucher specimens (No.
LD-8 and M-38) were deposited in the Herbarium of
Chengdu Institute of Biology, CAS.
3 Extraction and Isolation
Sample A: The air-dried and powdered sample A (5 kg)
was percolated with 95% ethanol (25 L × 3, each 7 days) at
room temperature. After evaporating the solvents under re-
duced pressure at 50°C, a residue (400 g) was obtained. The
residue was directly separated over silica gel column eluted
with petroleum ether-acetone (20: 1, 5: 1, V/V) to afford
three fractions, HA (50.3 g), HB (32.7 g), and HC(27.5 g),
and eluted with CHCl3-MeOH (10: 1 to 2: 1) to give HD
(5.2 g), HE (3.8 g), HF (41.3 g), HG (50.7 g), HI (61.3 g) and
HJ (107.7 g). In the mean time, compounds 1 (1 200 mg) and
2 (1 800 mg) were precipitated from HD and HE.
HB was divided into fractions HBA (5 g) and HBB (3
g) by silica gel column eluted with petroleum ether-acetone
(10: 1). HBB was further separated over silica gel column,
eluted with petroleum ether-acetone (10: 1) to give fraction
HBBA (100 mg) and HBBB (290 mg). HBBA was sepa-
rated over silica gel column, eluted with petroleum
ether-acetone (10: 1) to afford compound 3 (10 mg).
Compound 4 (20 mg) was obtained by recrystallizing the
precipitate of HBBB in petroleum ether-acetone (30: 1).
The chlorophyll in fraction HC (20 g) was removed
with MCI Gel column, eluted with methanol-water (90: 10,
100: 0) to yield CA (8 g), CB (2 g) and CC (4 g). CA was
separated over Sephadex LH-20 column, eluted with pure
methanol to give CAA (270 mg), CAB (110 mg) and CAC
(200 mg). CAC (100 mg) was further separated over silica
gel column, eluted with petroleum ether-acetone (30: 1) to
yield compounds 5 (7 mg) and 6 (4 mg). CB was subjected
to Sephadex LH-20 column, eluted by pure MeOH to yield
compound 7 (5 mg). CC (2.0 g) was separated over silica
gel column, eluted with petroleum ether-acetone (20: 1, 5:
1) and fractions CCA (40 mg), CCB (100 mg) and CCC (2
g) were obtained. CCB was separated on silica gel column
eluted with petroleum ether-acetone (10: 1) and compounds
8 (6 mg) and 9 (8 mg) were obtained.
Sample B: The air-dried and powdered sample B (3.2
kg) was percolated with 95% ethanol (30 L × 4, each 10
days) at room temperature. After evaporating the solvents
under reduced pressure at 50 °C, a residue (203 g) was ob-
tained. The residue was suspended in H2O (1.5 L) and frac-
tionated successively with petroleum ether (1.5 L × 2),
EtOAc (1.5 L × 5) and n-BuOH (1.5 L × 3) to afford petro-
leum ether fraction (16 g), EtOAc fraction (85 g) and
n-BuOH fraction (54 g). This fraction showed PTP1B in-
hibitory activity with an IC50 of 4.63 g·mL−1. The EtOAc
fraction was thus divided into fractions E1 (490 mg), E2 (1
g), E3 (2 g), E4 (1.9 g), E5 (3.6 g), E6 (1.3 g), and E7 (1.65
g) by C18 column (8 cm × 40 cm, 1 kg) eluted gradiently
with methanol-water (40: 100 to 100: 100, each 3 000
mL).
E1 was separated over a Sephadex LH-20 column (30
cm × 53 cm, 200 mL) eluted with MeOH (4 000 mL) to
afford four fractions, E1a (30mg), E1b (210 mg), E1c (190
mg) and E1d (28 mg). Fraction E1b was purified by
semi-preparative HPLC (MeOH: H2O = 30: 100) to yield
compound 10 (5 mg).
The separation of E2 over Sephadex LH-20 column
eluted with MeOH afforded three fractions, E2a (300 mg),
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18 Chin J Nat Med Jan. 2010 Vol. 8 No. 1 2010 年 1 月 第 8 卷 第 1 期

E2b (400 mg) and E2c (240 mg). E2a was further separated
over silica gel column, eluted with CHCl3-MeOH (50: 1)
to give fraction E2a2 (20 mg). E2a2 was purified by
semi-preparative HPLC (MeOH-H2O = 45: 55) to yield
compound 11 (8 mg).
E5 was subjected to silica gel column, using petroleum
ether-acetone (6: 1) to yield compound 12 (15 mg).
E6 was separated over Sephadex LH-20 column
eluted with MeOH to yield compound 13 (14 mg).
Compound 14 (19 mg) was crystallized from the
MeOH-H2O solution of E7.
4 PTP1B Assay
PTP1B inhibitory activity was determined with the
method of Seto[9].
5 Preliminary Screening of sample A
Fractions HC, HE, HF, HG and HI from sample A
were tested as ACE, H+-K+ ATP enzyme/proton pump,
α-glucosase/maltosase, sucrosase/amylase and PTP1B in-
hibitors. Five samples showed different PTP1B inhibitory
activities in vitro (Table 1).
Table 1 In vitro PTP1B inhibitory activities of the frac-
tions HC, HE, HF, HG and HI from sample A (μg·mL−1)
50 25 12.5 6.25
HC +++ +++ +++ +
HE +++ +++ ++ +
HF +++ +++ +++ +
HG ++ ++ + -
HI + - - -
6 Results and Discussion
Two samples of H. japonica (Thunb.) Dietr. were
collected from Luding County and Pingwu County, Sichuan,
China, respectively. Both of them showed PTP1B inhibitory
activities. Nine compounds were isolated from sample A
and five from sample B. Except compounds 13 and 14[3], all
the other compounds are first isolated from Helwingia jap-
onica (Thunb.) Dietr.. The compounds isolated are com-
pletely different. The difference may be derived from the
different growing environment. The medium polar fractions
of 95% ethanol in the two samples showed PTP1B inhibi-
tory activity. The compounds in the bioactive fractions are
thoroughly different. It is reported that betulinic acid (5) is
a PTP1B inhibitor[8]. Considering the low yield of betulinic
acid, the PTP1B inhibitory components in this plant need to
be studied further with not only some novel separating
methods with higher accuracy, but also different biological
testing model with boarder representative.
7 Identification
β-Sitosterol (1): Colorless needles, mp 137.2-139.5
°C (CHCl3). It was identified by comparing it with
authentic sample on TLC (CHCl3-acetone, V/V 20 : 1, Rf
0.50; petroleum ether-acetone, V/V 10 : 1, Rf 0.30;
petroleum ether-AcOEt, V/V 8 : 1, Rf 0.20) and co-mp[10].
Daucosterol (2): White powder. It was identified by
comparing it with authentic sample on TLC (CHCl3-acetone,
V/V 1 : 1, Rf 0.10; CHCl3-MeOH, V/V 6 : 1, Rf 0.45).
Lupenol (3): Colorless needles, mp 241.8-216.1°C
(acetone). It was identified by comparing it with authentic
sample on TLC (CHCl3-acetone, V/V 20: 1, Rf 0.65; petro-
leum ether-acetone, V/V 10: 1, Rf 0.40; petroleum
ether-AcOEt, V/V 8: 1, Rf 0.40).
Betulin (4): White powder, mp 214-216 °C,
[α] 20D +18.2° (c 0.49, CHCl3); IR (KBr) υmax: 3 436, 1 637,
1 457, 1 385, 1 262, 1 104 cm−1; 1H NMR (600 MHz,
CDCl3) δ: 4.68 (1H, br s, H-29a), 4.58 (1H, br s, H-29b),
3.79 (1H, d, J = 10.9 Hz, H-28a), 3.33 (1H, d, J = 10.9 Hz,
H-28b), 3.18 (1H, dd, J = 11.3, 4.6 Hz, H-3), 2.38 (1H, td,
J = 10.4, 5.6 Hz, H-19), 1.68 (3H, s, H-30), 1.02 (3H, s,
H-27), 0.98 (3H, s, H-26), 0.97 (3H, s, H-23), 0.83 (3H, s,
H-25), 0.76 (3H, s, H-24); 13C NMR (150 MHz, CDCl3) δ:
150.5 (C-20),109.7 (C-29), 79.0 (C-3), 60.6 (C-28), 55.3
(C-5), 50.9 (C-9), 48.8 (C-19),47.8 (C-8), 47.8 (C-18),42.7
(C-14), 40.9 (C-8), 38.9 (C-1), 38.7 (C-4), 37.3 (C-10), 37.2
(C-13), 34.2 (C-7), 34.0 (C-22), 29.8 (C-21), 29.2 (C-16),
28.0 (C-23), 27.4 (C-2), 27.1 (C-15), 25.2 (C-12), 20.8
(C-11), 19.1 (C-30), 18.3 (C-6), 16.1 (C-25), 16.0 (C-26),
15.4 (C-24), 14.8 (C-27). The IR, 1H, and 13C NMR data
were in accordance with those reported[11].
Betulinic acid (5): White powder, mp 316-318 °C,
ESI-MS m/z 455 [M－H]-, 911 [2M－H]- (negative mode),
[α] 20D +49° (c 0.24, MeOH); IR (KBr) υmax: 3 488, 2 964, 2
929, 1 688, 1 455, 1 186, 1 045 cm−1; 1H NMR (600 MHz,
C5D5N) δ: 4.69 (1H, d, J = 2.0 Hz, H-29β), 4.57 (1H, br s,
H-29α), 4.06 (1H, dd, J = 2.7 Hz, H-3), 1.68 (3H, s, H-30),
1.32 (3H, s, H-27), 1.06 (3H, s, H-26), 0.96 (3H, s, H-23),
0.79 (3H, s, H-25), 0.78 (3H,s, H-24); 13C NMR (150 MHz,
C5D5N) δ: 182.6 (C-28), 151.0 (C-20), 109.3 (C-29), 70.8
(C-3), 49.7 (C-17), 49.0 (C-5), 48.3 (C-9), 48.0 (C-19), 47.5
(C-7), 43.0 (C-14), 42.9 (C-8), 40.9 (C-4), 40.0 (C-1), 38.1
(C-13), 37.8 (C-10), 35.6 (C-22), 34.2 (C-7), 33.8 (C-16),
27.4 (C-15), 26.4 (C-21), 25.2 (C-23), 24.1 (C-2), 22.6
(C-12), 21.1 (C-11), 19.6 (C-30), 19.3 (C-6), 18.0 (C-26),
15.9 (C-25), 14.5 (C-24), 13.4 (C-27). The IR, 1H, and 13C
NMR data were in accordance with those reported[12, 13].
Glycerol monopalmitate (6): White amorphous
powder, mp 71-72 °C, ESI-MS m/z 353 [M + Na]+, 381 [M
+ K]+ (positive mode); IR (KBr) υmax: 3 433, 2 928, 1 693, 1
631, 1 285, 710 cm−1, 1H NMR (600 MHz, CDCl3) δ: 4.22
(1H, dd, J = 11.6, 4.5 Hz, H-1α), 4.17 (1H, dd, J = 11.6, 4.5
Hz, H-1β), 3.94 (1H, m, H-2), 3.71 (1H, m, H-3α), 3.62 (1H,
m, H-3β); 13C NMR (150 MHz, CDCl3) δ: 174.3 (C-1),
70.3 (C-2), 65.2 (C-3), 63.4 (C-1), 34.2 (C-2), 31.9 (C-14),
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2010 年 1 月 第 8 卷 第 1 期 Chin J Nat Med Jan. 2010 Vol. 8 No. 1 19

29.1-29.7 (C-4-13), 24.9 (C-3), 22.7 (C-15). 14.3 (C-16).
The IR, 1H, and 13C NMR data were in accordance with
those reported[14].
Cinnamic acid (7): White powder, mp 131-133 °C,
ESI-MS m/z: 295 [M+K]+ (positive mode), IR (KBr) υmax:
3 433, 2 928, 1 693, 1 631, 1 285, 710 cm−1, 1H NMR (600
MHz, CDCl3) δ: 7.81 (1H, d, J = 16.0 Hz, H-2), 7.57 (2H,
m, H-5, 9), 7.42 (3H, m, H-6, H-7, and H-8), 6.48 (1H, d, J
= 16.0 Hz, H-3); 13C NMR (150 MHz, CDCl3) δ: 172.1 (C-1),
147.1 (C-4), 134.1 (C-2), 130.7 (C-3), 129.0 (C-9), 129.0
(C-5), 128.4 (C-6), 128.4 (C-8), 117.3 (C-7). The IR, 1H, and
13C NMR data were in accordance with those reported[15].
Stignast-4-en-6β-3-one (8): White powder, ESI-MS
m/z 451 [M + Na]+, 467 [M + K]+ (positive mode),
[α] 20D +9.0° (c 0.14, CDCl3); IR (KBr) υmax: 3 429, 2 958, 2
868, 1 689, 1 467, 1 193, 1 018 cm−1; EtOHmaxUVλ (logε) nm:
238 (4.11); 1H NMR (600 MHz, CDCl3) δ: 5.82 (1H, s,
H-4), 4.35 (1H, m, H-6), 1.38 (3H, s, H-19), 0.95 (3H, d, J
= 9.4 Hz, H-21), 0.85 (3H, t, J = 7.4 Hz, H-29), 0.84 (3H, d,
J = 7.6 Hz, H-26), 0.82 (3H, d, J = 6.8 Hz, H-27), 0.74 (3H,
s, H-18); 13C NMR (150 MHz , CDCl3) δ: 200.4 (C-3),
168.4 (C-5), 126.3 (C-4), 73.3 (C-6), 56.1 (C-17), 55.9
(C-14), 53.6 (C-9), 45.8 (C-24), 42.5 (C-13), 39.6 (C-12),
38.6 (C-7), 38.0 (C-10), 37.1(C-1), 36.1 (C-20), 34.3(C-2),
33.9 (C-22), 29.2 (C-25), 28.2 (C-16), 29.7 (C-8), 26.1
(C-23), 24.2 (C-15), 23.1 (C-28), 21.0 (C-11), 19.8 (C-26),
19.0 (C-27), 18.7 (C-21), 19.5 (C-19), 12.0 (C-18), 12.0
(C-29). The UV, 1H, and 13C NMR data were in accordance
with those reported[16].
Stignast-4-en-6α-3-one (9): White powder, ESI-MS
m/z 429 [M + H]+, 451 [M + Na]+, 467 [M + K]+ (positive
mode), [α] 20D +20.0° (c 0.15, CDCl3), IR (KBr) υmax: 3 429,
2 958, 2 868, 1 689, 1 467, 1 193, 1018 cm−1;
EtOH
maxUVλ (logε) nm: 243 (4.21);
1H NMR (600 MHz, CDCl3)
δ: 6.18 (1H, d, J = 1.8 Hz, H-4), 4.35 (1H, m, H-6), 1.19
(3H, s, H-19), 0.95 (3H, d, J = 6.7 Hz, H-21), 0.85 (3H, t, J
= 7.4 Hz, H-29), 0.84 (3H, d, J = 7.6 Hz, H-26), 0.81 (3H, d,
J = 6.8 Hz, H-27), 0.72 (3H, s, H-18); 13C NMR (150 MHz ,
CDCl3) δ: 203.0 (C-3), 157.6 (C-5), 119.6 (C-4), 68.7 (C-6),
56.0 (C-17), 55.6 (C-14), 53.7 (C-9), 45.8 (C-24), 41.5
(C-13), 39.4 (C-12),39.4 (C-7), 39.1 (C-10), 36.3(C-1), 36.1
(C-20), 34.2 (C-2), 33.9 (C-22), 29.2 (C-25), 28.1 (C-16),
33.7 (C-8), 26.1 (C-23), 24.2 (C-15), 23.0 (C-28) 21.0
(C-11), 19.8 (C-26), 19.0 (C-27), 18.7 (C-21), 18.0 (C-19),
12.0 (C-18), 12.0 (C-29). The UV, 1H, and 13C NMR data
were in accordance with those reported[16].
p-Menth-2-en-1β, 4β, 8-triol (10): Colorless oil,
[α] 20D –0.11° (c 0.82, CHCl3); ESI-MS (positive mode) m/z
209 ([M + Na ]+), 225 ([M + K ]+); HRESI-MS (positive
mode) m/z 209.116 5 ([M + Na ]+, calcd. for C10H18Na1O3,
209.116 5); 1H NMR (600 MHz, CDCl3) δ: 5.85 (1H, d, J =
11.0 Hz, H-2), 5.80 (1H, d, J = 11.0 Hz, H-3), 1.94 (1H, dt,
J = 14.2, 4.2 Hz, H-5), 1.78 (2H, m, H-5,H-6), 1.68 (1H, m,
H-6), 1.26 (3H, s, H-10), 1.24 (3H, s, H-9), 1.16 (3H, s,
H-7); 13C NMR (150 MHz, CDCl3) δ: 138.8 (C-3), 128.1
(C-2), 74.3 (C-4), 73.6 (C-1), 69.5 (C-8), 34.9 (C-5), 29.3
(C-6), 26.7 (C-7), 24.5 (C-9), 24.1 (C-10). The optical rota-
tion, MS and NMR data were identical to those reported[17].
Blumenol A (11): Amorphous powder (MeOH),
[α] 25D +99.8° (c 0.65, CHCl3);
MeOH
maxUVλ (logε) nm: 237
(3.99); IR (KBr) υmax: 3 350, 2 900, 1 680, 1 350 cm−1; 1H
NMR (600 MHz, CDCl3) δ: 5.90 (1H, br s, H-4), 5.85 (1H,
dd, J = 15.7, 5.1 Hz, H-8), 5.79 (1H, d, J = 15.7 Hz, H-7),
4.41 (1H, m, H-9), 2.45 (1H, d, J = 16.8 Hz, H-2α), 2.24
(1H, d, J = 16.8 Hz, H-2β), 1.90 (3H, br s, H-13), 1.30 (3H,
d, J = 6.3 Hz, H-10), 1.08 (3H, s, H-12), 1.02 (3H, s, H-11);
13C NMR (150 MHz, CDCl3) δ: 198.0 (C-3), 162.7 (C-5),
135.8 (C-7), 129.0 (C-8), 126.9 (C-4), 79.0 (C-6), 68.0
(C-9), 49.7 (C-2), 41.1 (C-1), 24.0 (C-12), 23.8 (C-10), 22.9
(C-11), 18.9 (C-13). The optical rotation, UV, IR, 1H, and
13C NMR data were in accordance with those reported[18].
2′, 3′, 4′, 5′, 6′-Pentahydroxychalcone (12): Yellow
powder, mp 126-129 °C, ESI-MS (positive mode) m/z 289
([M + H ]+); MeOHmaxUVλ (logε) nm: 354 (2.58); IR (KBr)
υmax: 3 500, 1 653, 1 602 cm−1, 1H NMR (600 MHz, CDCl3)
δ: 7.80 (1H, d, J = 16.0 Hz, H-β), 7.56 (2H, m, H-2 and
H-6), 7.41 (3H, m, H-3, H-4 and H-5), 6.46 (1H, d, J = 16.0
Hz, H-α); 13C NMR (150 MHz, CDCl3) δ: 192.0 (C=O),
152.5 (C-2), 152.5 (C-6), 146.4 (C-4), 142.1 (C-β), 135.1
(C-1), 130.4 (C-4), 130.1 (C-3), 130.1 (C-5), 129.1 (C-2),
129.1 (C-6), 128.4 (C-3), 128.4 (C-5), 126.4 (C-α), 100.6
(C-1). The MS, UV, IR and NMR data were identical to
those reported[19].
Apigenin 7-O-β-D-glucopyranoside (13): Yellow
powder, mp 173-175 °C, [α]
20
D －52.8° (c 0.91, DMF);
ESI-MS (negative mode) m/z 431 ([M－H]-), 1H NMR (600
MHz,DMSO-d6): δ 12.96 (1H, s, 5-OH), 10.35 (1H, s,
4΄-OH), 7.95 (2H, d, J = 8.4 Hz, 2΄ and 6΄-H), 6.94 (2H, d,
J = 8.4 Hz, 3΄ and 5΄-H), 6.86 (1H, s, 3-H), 6.83 (1H, br s,
8-H), 6.44 (1H, br s, 6-H), 5.10 (1H, d, J = 7.4 Hz, 1΄΄-H).
The optical rotation, mp, ESI-MS and NMR data were in
accordance with those reported[20, 21].
Luteolin 7-O-β-D-glucopyranoside (14): Light yel-
low powder, mp 250-251 °C, [α] 20D －27.6° (c 1.0, pyri-
dine), ESI-MS (negative mode) m/z 447 ([M－H]-); 1H
NMR (600 MHz, DMSO-d6): δ 12.98 (1H, s, 5-OH), 8.59
(1H, s, 4΄-OH), 7.46 (1H, d, J = 8.4 Hz, 6΄-H), 7.44 (1H, br
s, 2΄-H), 6.92 (1H, d, J = 8.4 Hz, 5΄-H), 6.80 (1H, br s, 8-H),
6.78 (1H, s, 3-H), 6.46 (1H, br s, 6-H), 5.10 (1H, d, J = 7.4
Hz, 1΄΄-H). The optical rotation, mp, ESI-MS and NMR
data were identical to those reported[21, 22].
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27-28

青荚叶的化学成分
夏立子 1,2, 周 敏 3, 肖艳华 1, 李国友 1, 陈晓珍 1, 张国林 1*
1 中国科学院成都生物研究所, 成都 610041
2 中国科学院研究生院, 北京 100039
3 成都地奥集团, 成都 610041
【摘 要】 目的：为阐明青荚叶(Helwingia japonica (Thunb.) Dietr.) 地上部分 95%乙醇提取物中具有蛋白酪氨酸磷酸
酯酶 1B(protein tyrosine phosphatase, PTP1B)抑制活性的化学成分。方法：分别从四川芦定县和平武县采得青荚叶样品 A
和 B, 运用多种层析手段进行分离纯化, 通过波谱数据和理化性质进行化合物的鉴定。结果：样品 A 的 95%乙醇提取物按
极性分段后, HC、HE、HF、HG 部分具有强的抑制 PTP1B 活性。从样品 A 的 95%乙醇提取物中分离并鉴定了 9 个化合
物(1-9)。样品 B 的 95%乙醇提取物的乙酸乙酯部分显示 PTP1B 抑制活性(IC50 4.63 g·mL-1), 从中分离得到 5 个化合物
(10-14)。鉴定它们为谷甾醇(1)、β-胡萝卜苷(2)、羽扇豆醇(3)、桦木醇(4)、桦木酸(5)、棕榈酸甘油酯(6)、桂皮酸(7)、6αH-4-
烯-3-酮-豆甾醇(8)、6αH-4-烯-3-酮-豆甾醇(9)、p-menth-2-en-1β, 4β, 8-triol (10)、 blumenol A (11)、2′, 3′, 4′, 5′, 6′-五羟基查
尔酮 (12) 、洋芹素 7-O-β-D-吡喃葡萄糖苷 (13)和木犀草素 7-O-β-D-吡喃葡萄糖苷 (14)。结论：除了化合物 13 与 14, 其
余均是在该植物中首次分离得到。据报道化合物 5 有 PTP1B 抑制活性, 但考虑其在植物中的含量, 青荚叶中的活性成分有
待进一步研究。
【关键词】 青荚叶; 化学成分; 蛋白酪氨酸磷酸酯酶 1B