Chemical constituents of the root of jasminum giraldii

Article abstract of DOI:10.3390/molecules18044766

Two new compounds, ethylconiferin (1) and (-)-lariciresinol 4-(6"'-O-cinnamyl-β-D-glucopyranoside) (2), along with the three known compounds (+)-pinoresinol (3), (+)-cycloolivil (4), nobiletin (5), were isolated from the root of Jasminum girialdii. All th

Full text of DOI:10.3390/molecules18044766

Molecules 2013, 18, 4766-4775; doi:10.3390/molecules18044766
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molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
Chemical Constituents of the Root of Jasminum giraldii
1
,2,†
3,†
1,2
1,2
1,2
1,2
Zhenggang Yue
, Hui Qin , Yuhua Li , Yang Sun , Zhipeng Wang , Tiehong Yang ,
1
,2
4
3,
1,2,
Li Liu , Minchang Wang , Feng Feng * and Qibing Mei
*
1
Department of Pharmacology, School of Pharmacy, the Fourth Military Medical University,
Xi’an 710032, Shaanxi, China
2
3
Collaborative Innovation Center for Chinese Medicine in Qingba Mountains,
the Fourth Military Medical University, Xi’an 710032, Shaanxi, China
Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009,
Jiangsu, China
4
Xi’an Modern Chemistry Research Institute, Xi’an 710032, Shaanxi, China
†
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed; E-Mails: fengfeng@cpu.edu.cn (F.F.),
qbmei@fmmu.edu.cn (Q.M.); Tel.: +86-139-5204-5795 (F.F.); Fax: +86-25-8327-1269 (F.F.);
Tel./Fax: +86-29-8477-9212 (Q.M.).
Received: 1 February 2013; in revised form: 26 March 2013 / Accepted: 12 April 2013 /
Published: 22 April 2013
Abstract: Two new compounds, ethylconiferin (1) and (−)-lariciresinol
4
-(6'''-O-cinnamyl-β-D-glucopyranoside) (2), along with the three known compounds
(
+)-pinoresinol (3), (+)-cycloolivil (4), nobiletin (5), were isolated from the root of
Jasminum girialdii. All these compounds were isolated for the first time from this source.
Their structures were elucidated on the basis of extensive spectroscopic analysis and
chemical methods. In addition, the in vitro cytotoxic activity of these compounds was
evaluated. The results showed that none of the compounds had any significant inhibitory
activities on the proliferation of HCT-116 and SW-620 cells.
Keywords: Jasminum giraldii; oleaceae; chemical constituents; cytotoxic activity

Molecules 2013, 18
4767
1
. Introduction
Jasminum giraldii Diels (Oleaceae), is an endemic plant which is found distributed at an altitude of
3
00–1,500 m, in valleys and shrubbery of the Qinba Mountains in China’s Shaanxi Province [1], Its
dried roots, named as “quan pi” in Chinese, are used as a traditional local herb for the treatment of
various diseases, such as fractures, traumatic injury and blood stasis, while some other species of this genus
are employed to treat dysmenorrhea, metritis, leucorrhoea, galactophoritis, puerperal infection, irregular
menstruation, hyperthermia, arthralgia, pain due to ischemia, dermatitis [2], fever, rheumatic pain [3],
dysenteric diarrhea, trachelopanus, eczema [4], giddiness, edema [5] in Traditional Chinese Medicine.
The chemistry and pharmacology of the ethanol extracts of the roots of J. giraldii have never been
systematically investigated. As part of a program to acquire and assess potent chemicals in several
traditional Chinese medicines for the preservation and treatment of colorectal cancer [6–9], an ethanolic
extract of the root of J. giraldii has been investigated. We describe herein the isolation and structural
elucidation of two new compounds 1,2 (Figure 1) along with three known compounds 3–5 [10–12].
Furthermore, the cytotoxic activity of these compounds against HCT-116 and SW-620 cells is reported
for the first time.
Figure 1. The structures of compounds 1–5.
2
. Results and Discussion
Compound 1 was obtained as a white powder, whose molecular formula C18
H
26
O
8
was indicated by
+
HR-TOF-MS at m/z 393.1525 [M+Na] (calcd for C18
H
26
O
8
Na, 393.1525), implying six degrees of
unsaturation. The max absorptions in the UV spectrum of 1 were observed at 290, 259, 213 nm (in
−1
CH
aromatic ring (1510, 1454 cm ), and olefinic (1649 cm ) functionalities. The H-NMR spectra of 1
Table 1) in CD OD showed signals attributed to an aromatic ring ABX coupled system at ä 7.15(1H,
3
OH). The IR spectrum of 1 showed absorption bands of hydroxyl (3496, 3444, 3367 cm ),
−1
−1
1
(
3
d, J = 8.35 Hz, H-5), 7.11(1H, d, J = 1.90 Hz, H-2) and 6.99 (1H, dd, J = 1.90, 8.35 Hz, H-6), together
with signals attributed to an aromatic methoxy group at ä 3.90 (3H, s), The presence of a
trans-arylpropenoxy unit was characterized by signals at ä 6.61 (1H, d, J = 15.9 Hz, H-7), 6.26 (1H, dt,
J = 6.15, 15.9 Hz, H-8) and 4.15 (2H, dd, J = 1.25, 6.15 Hz, H-9). Meanwhile, an ethoxyl unit was

Molecules 2013, 18
4768
indicated by signals of 3.60 (2H, q, J = 7.05 Hz, H-1'), 1.25(3H, t, J = 7.05 Hz, H-2'), In addition, a
doublet assignable to an anomeric proton at  4.93 (1H, d, J = 7.25 Hz, H-1''), together with partially
overlapped signals attributed to oxymethylene and oxymethine protons at  3.90 (1H, dd, H-6a''), 3.72
(1H, dd, J = 7.45, 12.0 Hz, H-6b''), 3.52 (1H, m, H-2''), 3.51 (1H, m, H-3''), 3.50 (1H, m, H-5''), 3.48
(
1H, m, H-4''), suggested that there was a glycosyl moiety with a  configuration in 1. Enzymatic
1
hydrolysis of 1 produced a sugar, which was identified as glucose by its H-NMR data and TLC
comparison with an authentic sugar sample. The glucose isolated from the hydrolysate gave a positive
2
D
0
13
optical rotation [α]
: +47.2, indicating that it was D-glucose [13]. The C-NMR and DEPT spectra of
1
showed carbon signals corresponding to the above units (Table 1). The HMBC correlation signals
(
Table 1 and Figure 2) of H-1'' (δ 4.93)/C-4 (δ 147.96) indicated that the glucosyl group was linked
H
C
to C-4, H-7 (δ
H
6.61)/C-1 (δ
C
C C
133.38), C-2 (δ 111.59), C-6 (δ 121.0) indicated that the
trans-arylpropenoxy unit was connected to the aromatic ring via a C-7C-1 bond, H-1' (δ
H
3.60)/C-9
1
(
δ 72.38) indicated that the ethoxyl unit was linked to C-9. Therefore, compound 1, whose H-NMR
C
1
3
and C-NMR were similar to those of methylconiferin [14], was named ethylconiferin.
Figure 2. Key HMBC correlations in compound 1.
1
13
Table 1. H-NMR (500 MHz) and C-NMR (125 MHz) spectral data and HMBC
Correlations of Compound 1 in CD OD.
3
NO.
1
δ
C
δ
H
(J, Hz)
HMBC
133.38
111.59
151.07
147.96
118.08
121.0
2
7.11 (1H, d, 1.9)
4, 6
3
4
5
7.15 (1H, d, 8.35)
6.99 (1H, dd, 1.9, 8.35)
6.61 (1H, d, 15.9)
1, 3
1, 2, 4, 5
1, 2, 6, 9
1, 9
6
7
133.55
126.13
72.38
8
6.26 (1H, dt, 6.15, 15.85)
4.15 (2H, dd, 1.25, 6.15)
3.60 (2H, q, 7.05)
1.25 (3H, t, 7.05)
4.93 (1H, d, 7.25)
3.52 (1H, m)
9
1, 1', 8
2', 9
1
2
'
'
66.78
15.59
1'
1
2
3
4
5
''
''
''
''
''
102.89
75.05
4
1'', 3''
2'', 4''
5''
78.00
3.51 (1H, m)
71.49
3.48 (1H, m)
78.38
3.50 (1H, m)
6
a'
b''
-OCH3
3.90 (1H, dd)
6
2.66
6
3.72 (1H, dd, 7.45, 12)
3.90 (3H, s)
3
56.87
3

Molecules 2013, 18
4769
Compound 2 was isolated as a white powder, whose molecular formula C H O was determined by
3
5
40 12
+
HR-TOF-MS at m/z 653.2589 [M+H] (calcd for C35
41
H O12, 652.2590), The UV absorption bands of 2
appeared at 278, 216 and 203 nm in CH OH. The IR spectrum of 2 showed the presence of hydroxyl
3
−
1
−1
−1
(
3498, 3458, 3384 cm ), conjugated-ester carbonyl (1714 cm ), α,β-unsaturated olefinic (1645, 1635 cm )
−1
1
and aromatic ring (1512, 1456 cm ) functionalities. The H-NMR data of 2 (Table 2) demonstrated
signals attributable to two pairs of 1,3,4-trisubstituted benzene ring signals at δ 7.04 (1H, d, J = 8.25 Hz,
H-5), 6.85 (1H, d, J = 1.75 Hz, H-2), 6.61 (1H, dd, J = 1.8, 8.25 Hz, H-6) and 6.90 (1H, d, J = 1.4 Hz,
H-2'), 6.78 (1H, m H-5'), 6.77 (1H, m, H-6'); and a set of cinnamyl proton signals at δ 7.61 (2H ,m, H-2'',
6
''), 7.43 (3H, m, H-3'', 4'', 5''), 7.68 (1H, d, J = 16 Hz, H-7''), 6.56 (1H, d, J = 16 Hz, H-8'') in the
aromatic region. In addition, resonances assignable to two aromatic methoxyl proton signals at δ 3.86
(
3H, s, 3'-OCH
3
), 3.85 (3H, s, 3-OCH
3
), and an anomeric proton signal at δ 4.88 (1H, d, H-1''') were
1
observed. Furthermore, in the H-NMR spectrum, an O-bearing methine proton signals at δ 4.72 (1H, d,
J = 6.65 Hz, H-7'), an aliphatic methylene proton signals at δ 2.85 (1H, dd, J = 4.7, 13.5 Hz, H-7a), 2.38
(1H, dd, 11.55, 13.3Hz, H-7b), two oxygenated methylene proton signals at δ 3.88(1H, dd, J = 6.5, 8.35 Hz,
H-9a), 3.62 (1H, m, H-9b), 3.76 (1H, m, H-9a'), 3.60 (1H, m, H-9b'), and two aliphatic methine proton
signals at δ 2.59 (1H, m, H-8), 2.31 (1H, m, H-8') were indicated. Besides the carbon resonances
1
3
corresponding to the above units, the C-NMR spectrum (Table 2) and the DEPT indicated the presence
of conjugated-ester carbonyl carbon signals at δ 168.37 (C-9''); a set of a β-glucose moiety carbon
signals at δ 102.87 (C-1'''), 77.95 (C-3'''), 75.61 (C-5'''), 75.03 (C-2'''), 72.13 (C-4'''), 64.96 (C-6'''). The
3 3
key HMBC correlations (Table 2 and Figure 3) from 3-OCH to C-3; from 3'-OCH to C-3'; from H-7 to
C-1, C-2, C-6, C-8 and C-9; from H-9 to C-7, C-7', C-8 and C-8'; from H-7' to C-1', C-2', C-6', C-8', C-9
and C-9'; from H-8' to C-1', C-7, C-7', C-8, C-9 and C-9'; and from H-9' to C-7, C-7', C-8 and C-8';
suggested that the aglycone should be lariciresinol [15]. Moreover, the HMBC correlations of H-1'''/C-4
confirmed that anomeric protons of β-glucose moiety was coupled with C-4. Meanwhile, the more
H
deshielded proton of H-6''' (δ 4.55 and 4.33) exhibited a HMBC correlation with C-9'', which confirmed
that the cinnamyl moiety was linked to C-6'''. In the NOESY spectrum of 2 (Figure 3), H-8' showed NOE
correlations with H-2', H-6' and H-8, this experiment confirmed that H-8 and H-8' are in
cis-configuration, meanwhile, H-7' and H-8' are in trans-configuration. Comparison of the NMR data of
(
−)-lariciresinol 4'-(6''-O-feruloyl-β-D-glucopyranoside) [16] and compound 2 indicated they had very
similar aglycone and sugar moieties, the only difference being the presence of a cinnamyl moiety instead
of a feruloyl group. Compound 2 was hydrolysed by dilute acid, and the hydrolysate was identified to be
D-glucose by the same way as described in the case of 1, Therefore the structure of 2 was determined to
be (−)-lariciresinol 4-(6'''-O-cinnamyl-β-D-glucopyranoside).
Figure 3. Key HMBC and NOESY correlations in compound 2.

Molecules 2013, 18
4770
1
13
Table 2. H-NMR (500 MHz) and C-NMR (125 MHz) spectral data and HMBC
3
correlations of compound 2 in CD OD.
No.
1
δ
C
δ
H
(J, Hz)
HMBC
137.19
114.46
150.95
146.21
118.42
122.34
2
6.85 (1H, d, 1.75)
1, 3, 4, 6, 7
3
4
5
7.04 (1H, d, 8.25)
1, 2, 3, 4
2, 3, 4, 5, 7
1, 2, 6, 8, 9
1, 2, 6, 8
6
6.61 (1H, dd, 1.8, 8.25)
2.85 (1H, dd, 4.7, 13.5)
2.38 (1H, dd, 11.55, 13.3)
2.59 (1H, m)
7
a
3
3.85
43.66
3.53
7
b
8
9
a
3.88 (1H, dd, 6.5, 8.35)
3.62 (1H, m)
7, 7', 8'
7
9
b
7, 7', 8, 8'
1
'
'
'
'
'
'
'
'
135.87
110.78
149.14
147.2
2
6.90 (1H, d, 1.4)
1', 3', 4', 6', 7'
3
4
5
6
7
8
116.15
119.93
84.11
6.78 (1H, m)
6.77 (1H, m)
4.72 (1H, d, 6.65)
2.31 (1H, m)
3.76 (1H, m)
3.60 (1H, m)
1', 3', 4'
2', 8'
1', 2', 6', 8', 9, 9'
1', 7, 7', 8, 9, 9'
7, 7', 8, 8'
54.15
9
a'
b'
''
6
0.59
9
7, 7', 8, 8'
1
135.90
129.47
130.28
131.75
146.59
118.96
168.37
102.87
75.03
2
3
'',6''
'',5''
7.61 (2H, m)
7.43 (2H, m)
4'', 7''
1''
4
7
8
9
''
''
7.43 (1H, m)
1'', 3'', 5''
1'', 2'', 6'', 8'', 9''
1'', 9''
7.68 (1H, d, 16)
6.56 (1H, d, 16)
''
''
1
2
3
4
5
'''
'''
'''
'''
'''
4.88 (1H, d)
3.53 (1H, m)
3.52 (1H, m)
3.43 (1H, t)
4
1''', 4'''
2''', 4'''
3''', 5''', 6'''
1''', 4''', 6'''
5''', 9''
5''', 9''
3
77.95
72.13
75.61
3.72 (1H, m)
4.55 (1H, dd, 2.2, 11.75)
4.43 (1H, dd, 7.4, 11.65)
3.85
6
a'''
b'''
-OCH3
’-OCH3
6
4.96
6
3
56.85
56.55
3
3.86
3'
The known compounds 3–5 were identified by comparison of their spectroscopic data with literature
values. HCT-116 and SW-620 cells were treated with three different concentrations of compounds for
2
4 h. The antiproliferative effects of the five compounds on HCT-116 and SW-620 cells are shown in
Table 3. None of the tested compounds had any significant effects on proliferation of HCT-116 cells and
SW-620 cells at the tested concentrations of 1–10 M.

Molecules 2013, 18
Table 3. Activity of compounds 1–5 on proliferation of the HCT-116 and SW-620 cells.
4771
HCT-116 cells
SW-620 cells
−
4
−5
−6
−4
−5
−6
1
0
mol/L
Inhibitory rate
%)
5.383 ± 2.187 **
10 mol/L
10 mol/L
10 mol/L
10 mol/L
10 mol/L
Inhibitory rate
(%)
Sample
Inhibitory rate
(%)
Inhibitory rate
(%)
Inhibitory rate
(%)
Inhibitory rate
(%)
(
1
16.150 ± 0.504 ** 13.329 ± 0.367 **
6.967 ± 0.262 **
5.026 ± 0.713 **
3.141 ± 1.101 **
8.605 ± 0.745 **
2.894 ± 0.960 **
3.769 ± 0.799 **
0
1.218 ± 1.016 **
2.894 ± 0.461 **
5.863 ± 0.287 **
6.035 ± 0.605 **
2.646 ± 1.577 **
0
2
13.049 ± 1.607 ** 14.685 ± 1.615 ** 11.671 ± 0.941 ** 12.260 ± 0.443 **
3
12.086 ± 1.404 ** 13.157 ± 0.748 **
14.750 ± 2.171 ** 6.288 ± 1.735 **
19.079 ± 0.440 ** 13.868 ± 0.373 **
5.943 ± 0.931 **
3.725 ± 0.914 **
2.972 ± 0.734 **
0
24.119 ± 1.128 **
24.481 ± 0.184 **
26.499 ± 0.453 **
0
4
5
Control
0
0
The data are expressed as mean ± SD of three independent experiments. (** p < 0.01 vs. control).
3
. Experimental
.1. General
UV spectra were taken with a HALO DB-20R UV-VIS spectrophotometer. Optical rotations were
3
recorded on a PerkinElmer Model 343 polarimeter. The IR spectra were recorded on a Shimadzu
FTIR-8400S instrument. ESI-MS was performed on s Waters Quattro Premier instrument. The
HR-ESIMS spectra was taken on an Agilent Technologies 6550 Q-TOF. 1D and 2D NMR spectra were
recorded on a Bruker-AVANCE500 instrument with TMS as an internal standard. The analytical HPLC
was performed on a Waters 2695 Separations Module coupled with a 2996 Photodiode Array Detector and a
ODS-3 column (4.6 × 250 mm, 5 ìm particles, Inertsill, Tokyo, Japan). Semipreparative HPLC was
performed on a system comprising a Shimadzu LC-6AD pump equipped with a SPD-20A UV detector
and a Ultimate XB-C18 (10 × 250 mm, 5 μm particles) or Allsphere ODS-2 (10 × 250 mm, 5 μm
particles). Sephadex LH-20 was purchased from GE Healthcare Bio-Sciences AB (Uppsala, Sweden).
MCI GEL was from Mitsubishi Chemical Corporation (Tokyo, Japan). C-18 (40–75 μm) silicagel was
purchased from SiliCycle Corporation (Quebec, Canada). D101 was from Sunresin New Materials Co.
Ltd. (Xi’an, China). Silica gel was purchased from Qingdao Haiyang Chemical Group Corporation
(Qingdao, China).
3.2. Plant Materials
The root of Jasminum giraldii Diels. was collected on October in 2009 in Shanxi, China, and
authenticated by Prof. Yaowu Guo. A voucher specimen has been deposited in the Department of
Collaborative Innovation Center for Chinese Medicine in Qingba Mountains, School of Pharmacy the
Fourth Military Medical University.
3.3. Extraction and Isolation
The root of J. giraldii (20 kg) was powdered and extracted with EtOH (100 L, 3 h × 4) under reflux.
Evaporation of the solvent under reduced pressure gave the EtOH extract. The combined extract was
applied to D101 and eluted with EtOH-H O to obtain the 95% EtOH eluate (15 L). The eluate was
2

Molecules 2013, 18
4772
evaporated under vacuum and freeze dried to yield a powder (895 g), which was suspended in H O and
2
partitioned successively with petroleum ether, EtOAc and n-BuOH. The EtOAc fraction was subjected
to silica-gel column chromatography eluting with CHCl
Fr.1–Fr.33). Fr.28 was further separated by flash chromatography over MCI, eluting with a gradient of
MeOH (0–100%) in H O to obtain seven fractions (Fr.28-1–Fr.28-7). Repeated chromatography of
3
-MeOH (100:0 to 1:1) to obtain 33 fractions
(
2
Fr.28-1 with silica gel (MeOH/CHCl
prep-HPLC gave compounds 1 (8 mg), 2 (26 mg), 4 (28 mg), Fr.21 was subjected to silica gel
200–300 mesh) column chromatography and eluted with CHCl /MeOH (100:0–50:1) to obtain five
3 3
, 30:1–5:1), Sephadex LH-20 (CHCl /MeOH, 1:1), prep-TLC and
(
3
fractions (Fr.21-1–Fr.21-5). Fr.21-5 was further separated repeated with silica gel column (petroleum
ether/acetone, 8:1–5:1), Sephadex LH-20 (CHCl /MeOH, 1:1), prep-TLC and prep-HPLC to afford
3
compounds 3 (120 mg) and 5 (5 mg).
3.4. Spectral Data
2
D
0
Ethylconiferin (1): white powder; C18
H
26
O
8
; [α]
3
: −19.76 (c 0.1, CH OH); UVmax (MeOH) λmax 290,
2
1
59 and 213 nm; IR (KBr) νmax 3496, 3444, 3367, 2979, 2939, 2867, 1649, 1510, 1454, 1128, 1089,
−1
+
1
8
047, 1027 cm ; HR-TOF-MS m/z 393.1525 [M+Na] (calcd for C18H26NaO ; 393.1525); H-NMR
1
3
3 3
(500 MHz, CD OD) and C-NMR (125 MHz, CD OD) data, see Table 1.
2
D
0
(−)-Lariciresinol 4-(6'''-O-cinnamyl-β-D-glucopyranoside) (2): white powder; C35
40
H O12; [α]
: −10.91
(
c 0.1, MeOH); UVmax (MeOH) λmax 278, 216 and 203 nm; IR(KBr) νmax 3498, 3458, 3384, 2925, 2854,
−1
1
6
714, 1645, 1635, 1512, 1456, 1269, 1226, 1161, 1124, 1105, 1072, 1035 cm ; HR-TOF-MS m/z
+
1
13
53.2590 [M+H] (calcd for C35
H
41
O
8
; 653.2590); H-NMR (500 MHz, CD
3
OD) and C-NMR (125 MHz,
CD
3
OD) data, see Table 2.
2
D
0
(+)-Pinoresinol (3): yellow powder; C20
H
22
O
6
; [α]
: +46.0 (c 0.1, MeOH); UVmax (MeOH) λmax 280,
2
1
6
5
3
31 and 207 nm; IR (KBr) νmax 3504, 3438, 3379, 2958, 2939, 2860, 1604, 1514, 1460, 1438, 1271,
−1
 1
029 cm ; ESI-MS m/z 357 [M−H] ; H-NMR (500 MHz, Me CO-d ) δ 7.47 (2H, s, 4-OH and 4'-OH),
2
6
.99 (2H, d, J = 1.7 Hz, H-2, 2'), 6.84 (2H, dd, J = 1.7, 8.1 Hz, H-6, 6'), 6.79 (2H, d, J = 8.1 Hz, H-5,
'),4.67 (2H, d, J = 4.15 Hz, H-7, 7'), 4.20 (2H, dd, J = 6.95, 8.95 Hz, H-9a, 9'a), 3.84(6H, s, 3-OCH and
'-OCH ), 3.81 (2H, dd, J = 3.7, 9.1 Hz, H-9b, 9'b), 3.08 (2H, m, H-8, 8'); C-NMR (125 MHz,
3
1
3
3
Me
2
CO-d
6
) δ 148.40 (C-3, 3'), 146.94 (C-4, 4'), 134.27 (C-1, 1'), 119.69 (C-6, 6'), 115.62 (C-5, 5'),
and 3'-OCH ), 55.32 (C-8, 8').
1
10.67 (C-2, 2'), 86.72 (C-7, 7'), 72.29 (C-9, 9'), 56.33 (3-OCH
3
3
2
D
0
(
+)-Cycloolivil (4): yellow powder; C20
H
24
O
7
; [α]
: +43.2 (c 0.1, MeOH); UVmax (MeOH) λmax 283, 230
and 204 nm; IR (KBr) νmax 3517, 3444, 3313, 3201, 2939, 2900, 2835, 1699, 1598, 1517, 1465, 1446,
−
1

1
1
433, 1373, 1269, 1236, 1215, 1149, 1124, 1022, 881, 858, 838, 811 cm ; ESI-MS m/z 375 [M−H] ;
3
H-NMR (500 MHz, CD OD) δ 6.76 (1H, d, J = 8.0 Hz, H-5), 6.76 (1H, d, J = 1.6 Hz, H-2), 6.67 (1H,
dd, J = 1.7, 7.95 Hz, H-6), 6.63 (1H, s, H-5'), 6.18 (1H, s, H-2'), 4.03 (1H, d, J = 11.65 Hz, H-7), 3.80
3H, s, OCH ), 3.77 (2H, m, H-9a, 9'a), 3.74 (3H, OCH ), 3.59 (2H, m, H-9b, 9'b), 3.23 (1H, d, J = 16.65
Hz, H-7'a), 2.62 (1H, d, J = 16.65 Hz, H-7'b), 2.04 (1H, m, H-8); C-NMR (125 MHz, CD
49.27 (C-3), 147.64 (C-3'), 146.25 (C-4), 145.44 (C-4'), 138.62 (C-1'), 133.69 (C-1), 126.57 (C-6'),
(
3
3
1
3
3
OD) δ
1

Molecules 2013, 18
4773
1
23.71 (C-6), 117.48 (C-5), 116.15 (C-5'), 114.06 (C-2), 113.09 (C-2'), 75.10 (C-8'), 69.55 (C-9), 60.98
(
C-9'), 56.52 (OCH ), 56.49 (OCH ), 47.69 (C-8), 45.03 (C-7), 40.06 (C-7').
3
3
2
D
0
Nobiletin (5): white powder; C
9
H O
12 2
; [α] : +3.69 (c 0.1, MeOH); UVmax (MeOH) λmax 328, 269, 249
and 208 nm. IR (KBr) ν 2947, 2854, 1629, 1598, 1514, 1461, 1377, 1278, 1149, 1107, 1078, 1012,
max
−1
+
1
9
73, 838 cm ; ESI-MS m/z 403 [M+H] ; H-NMR (500 MHz, Me
2 6
CO-d ) δ 7.66 (1H, dd,
J = 2.15, 8.5 Hz, H-6'), 7.59 (1H, d, J = 2.05 Hz, H-2'), 7.15 (1H, d, J = 8.5 Hz, H-5'), 6.62 (1H, s, H-3),
4
.07 (3H, s, OCH ), 4.04 (3H, s, OCH ), 3.96 (3H, s, OCH ), 3.92 (3H,s, OCH ), 3.89 (3H, s, OCH ),
3 3 3 3 3
1
3
3 2 6
3.85 (3H, s, OCH ); C-NMR (125 MHz, Me CO-d ) δ 176.74 (C-4), 161.67 (C-2), 153.38 (C-4'), 152.4
(
(
C-7), 150.72 (C-3'), 149.22 (C-9), 148.64 (C-8), 145.15 (C-5), 139.24 (C-6), 124.85 (C-1'), 120.39
C-6'), 115.91 (C-10), 112.69 (C-5'), 110.01 (C-2'), 107.29 (C-3), 62.44, 62.37, 62.04, 61.98, 56.34,
5
3 3 3 3 3 3
6.31(5-OCH 6-OCH , 7-OCH , 8-OCH , 3'-OCH and 4'-OCH ,).
3
.5. Enzymatic Hydrolysis of 1
A solution of compound 1 in H O (3 mL) was hydrolysed with β-glucosidase (10 mg, Almonds Lot
2
1
264252, Sigma-Aldrich, St. Louis, MO, USA) at 37 °C for 24 or 36 h. The reaction mixture was
extracted with EtOAc for three times (3 × 3 mL) to yield the EtOAc exrract and H
removing the solvents. The aqueous phase of the hydrolysate of compound 1 was subjected to CC over
silica gel eluted with MeCN/H O (8:1) to yield glucose with positive optical rotations, and the value of
O). The solvent system MeCN/H O (6:1) was used for the TLC analysis of
glucose and authentic sugar samples.
2
O phase after
2
2
D
0
[α]
: was +47.2 (c 0.16, H
2
2
3
.6. Acid Hydrolysis of 2
A solution of compound 2 in 0.5 M H
2
SO
O, neutralized with BaCO
extracted with EtOAc for three times (3 × 3 mL) to obtained the EtOAc extract and H
removing the solvents. The aqueous layer of the hydrolysate of compound 2 was evaporated and to
4
(2.0 mL) was heated under reflux for 3 h. After cooling,
, then filtered. The solution was
O layer after
the reaction mixture was diluted with H
2
3
2
2
0
yield glucose with positive optical rotations, and the value of [α]
D
2
: was +42.5 (c 0.11, H O). The solvent
system MeCN/H O (6:1) was used for the TLC analysis of glucose and authentic sugar samples.
2
3
.7. Bioassay for Cytotoxic Activity
The human colon cell line HCT-116 and SW620 was obtained from the Shanghai Institute of Cell
Biology. The cells were maintained in DMEM medium supplemented with 10% FBS, penicillin (100 U/mL)
and streptomycin (100 mg/mL) at 37 °C in a humidified atmosphere with 5% CO . Cell proliferation of
2
HCT-116 cells was assessed by conducting colorimetric MTT cell proliferation assay. Briefly, a limited
number of growing cells were seeded into 96-well cell culture plate and were maintained for 24 h so that
they became attached to the bottom of the well. The medium was aspirated and new medium (200 µL)
−4
−5
−6
including three groups of concentrations (10 , 10 , 10 M) of compounds were added to parallel wells.
The cells were incubated at 37 °C in a humidified atmosphere for 24 h. Twenty µL of MTT solution was
added and the incubation continued for 4 h. The pure formazan product was then solubilized by 150 µL
DMSO. The plates were read at 570 nm using a microtiter plate reader.

Molecules 2013, 18
. Conclusions
Compounds 1, 2 were new glycosides and together with compounds 3–5 were isolated from the
4774
4
Jasminum genus for the first time. Compounds 1–5 were assayed for their cytotoxic activity, and the data
proved that none of the compounds had any significant inhibitory activities on proliferation of HCT-116
cells and SW-620 cells at the concentrations of 1–10 M.
Supplementary Materials
Supplementary materials can be accessed at: http://www.mdpi.com/1420-3049/18/4/4766/s1.
Acknowledgments
Financial support from the National Natural Sciences Foundation of China (NNSFC; Grant
No. 81072671), and the National Program on Key Basic Research Project of China (973 Program; Grant
No. 2010CB535002) is acknowledged.
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2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
http://creativecommons.org/licenses/by/3.0/).
(