2
50
J Nat Med (2010) 64:245–251
-
C H O : 479.1765). IR (KBr) cm : 3408, 2925, 1597,
1
4-Hydroxy-3-methoxyphenylacetonitrile (1a)
An orange solid. Positive HR FAB-MS (NBA) m/z:
2
0 31 13
1
3
1
516. C- and H-NMR: Table 1. Selected HMBC cor-
1
0
relations: H-7/C-1, C-2, C-6; OCH /C-3; H-6/C-4; H-1 /C-
3
0
0
0
?
-1
4
; H-1 /C-6 .
163.0631 ([M] , C
9
H
9
NO
390, 2925, 2252, 1603, 1518. C-NMR (DMSO-d6,
25 MHz) d: 21.9 (C-7), 55.6 (OCH ), 112.3 (C-2), 115.7
2
:163.0633). IR (Nujor) cm
:
1
3
3
1
Known compounds isolated
3
(
C-5), 119.7 (C-8), 120.5 (C-6), 121.6 (C-1), 146.0 (C-4),
1
2
5
147.8 (C-3). H-NMR (DMSO-d , 500 MHz) d: 3.75 (3H,
Vanilloloside (6) [16]: A white amorphous powder. [a]D
47.7° (c 0.50, MeOH). Positive HR FAB-MS (NBA) m/z:
6
s, OCH ), 3.85 (2H, s, H -7), 6.72 (1H, dd, J = 1.7,
3
-
2
?
16.1148 ([M] , C H O : 316.1158). IR (KBr) cm
-1
8.0 Hz, H-6), 6.75 (1H, d, J = 8.0 Hz, H-5), 6.87 (1H, d,
J = 1.7 Hz, H-2), 9.07 (1H, s, 4-OH). Selected HMBC
correlations: H-7/C-1, C-2, C-6, C-8; OCH /C-3; H-6/C-4;
3
:
1
4 20 8
1
433, 2922, 1597, 1514. C-NMR (DMSO-d , 125 MHz)
3
3
6
0
0
d: 55.6 (OCH ), 60.7 (C-6 ), 62.7 (C-7), 69.7 (C-4 ), 73.3
3
3
0
0
0
0
OH/C-3, C-4, C-5.
(
C-2 ), 76.9 (C-5 ), 77.0 (C-3 ), 100.2 (C-1 ), 111.1 (C-2),
1
15.2 (C-5), 118.6 (C-6), 136.4 (C-1), 145.3 (C-4), 148.8
1
(
C-3). H-NMR (DMSO-d , 500 MHz) d: 3.15 (1H, m,
4-Hydroxyphenylacetonitrile (4a)
6
0
0
H-4 ), 3.23 (1H, overlapped, H-2 ), 3.23 (1H, overlapped,
0
0
H-3 ), 3.26 (1H, overlapped, H-5 ), 3.43 (1H, ddd,
0
An orange solid. Positive HR FAB-MS (NBA) m/z:
?
-1
133.0534 ([M] , C H NO:133.0528). IR (Nujor) cm :
8 7
J = 5.8, 5.8, 12.1 Hz, H-6 a), 3.64 (1H, br dd, J = 4.6,
0
13
1
2.1 Hz, H-6 b), 3.73 (3H, s, OCH ), 4.40 (2H, d,
3
3326, 2925, 2252, 1610, 1516. C-NMR (DMSO-d6,
125 MHz) d: 21.5 (C-7), 115.6 (C-3), 115.6 (C-5), 119.7
(C-8), 121.1 (C-1), 129.2 (C-2), 129.2 (C-6), 156.8 (C-4).
0
J = 5.2 Hz, H-7), 4.85 (1H, d, J = 7.5 Hz, H-1 ), 5.09
(
1H, t, J = 5.2 Hz, 7-OH), 6.78 (1H, dd, J = 1.8,
.0 Hz, H-6), 6.92 (1H, d, J = 8.0 Hz, H-5), 7.01 (1H, d,
J = 1.8 Hz, H-2). Selected HMBC correlations: H-7/C-1,
1
8
H-NMR (DMSO-d , 500 MHz) d: 3.85 (2H, s, H -7), 6.75
6
2
(2H, d, J = 8.6 Hz, H-3 and H-5), 7.12 (2H, d, J = 8.6 Hz,
H-2 and H-6), 9.49 (1H, s, 4-OH). Selected HMBC
correlations: H-7/C-1, C-2, C-6, C-8; OH/C-3, C-4, C-5.
C-2, C-6; OCH /C-3; H-6/C-4; H-2/C-4; 7-OH/C-1, C-7;
3
0
H-1 /C-4.
3
,4-Dimethoxybenzyl alcohol 7-O-b-D-glucopyranoside
2
5
(
7) [16]: A white amorphous powder. [a] -39.9° (c 0.33,
Synthesis of 4-hydroxy-3-methoxyphenylacetonitrile (1a)
D
MeOH).
Lobetyolin (8) [17]: A brown syrup. [a]D -26.6° (c 1.89,
MeOH).
2
5
4-Hydroxy-3-methoxyphenylacetonitrile (1a) was synthe-
sized as described elsewhere but with slight modifications
[
18]. Briefly, a mixture of 4-hydroxy-3-methoxybenzyl
Enzymatic hydrolysis of 1–5
alcohol (1.23 g, 8.0 mmol) and NaCN (472 mg, 9.6 mmol)
in DMF (20 ml) was stirred under nitrogen at 120°C for
24 h. The reaction mixture was cooled and 0.4 ml of water
was added, the mixture was then alkalized with solid
NaOH to more than pH 10, followed by evaporation under
vacuum. Water (1 ml) was added and neutralized with
acetic acid. After the mixture was extracted with CHCl3,
Each glycoside (10.0 mg) was incubated with emulsin
(
b-glucosidase from almonds: G-0395: Lot No. 119H4029;
.8 units; Sigma, MO, USA) in McIlvaine buffer (pH 5.0)
6
at 37°C for 4 h. The reaction mixture was diluted with
H O and passed through a Sep-pak ODS cartridge
2
(
Waters, MA, USA), eluting with H O and MeOH, to
2
the extract was washed with H O three times and dried
2
afford an H O eluate and a MeOH eluate, respectively. In
2
over anhydrous Na SO , followed by evaporation under
2
4
the case of 1–4, the MeOH eluate was purified by reverse-
phase preparative HPLC (40% MeOH). Finally, 1a was
obtained as aglycone in the case of 1–3, and 4a and 5a
were obtained in the case of 4 and 5, respectively. The
vacuum. The extract (1.26 g) was purified by silica gel CC
[hexane–acetone (2:1)] and then ODS CC (30% MeOH) to
give 1a (315 mg, 1.9 mmol) as an orange solid.
H O eluate was analyzed by HPLC under the following
2
Synthesis of 4-hydroxyphenylacetonitrile (4a)
conditions: column, COSMOSIL Sugar-D; solvent, 70%
acetonitrile; flow rate, 1.0 ml/min; detectors, RI-2031 and
4-Hydroxyphenylacetonitrile (4a) was synthesized simi-
larly to 1a but with slight modifications. Briefly, a mixture
of 4-hydroxybenzyl alcohol (1.01 g, 8.15 mmol) and
NaCN (483 mg, 9.9 mmol) in DMF (25 ml) was stirred
under nitrogen at 120°C for 26 h. The reaction mixture was
cooled and 0.5 ml of water was added, then the mixture
OR-2090 plus. Consequently, the constituent of the H O
2
eluate was detected to be D-glucose (t 5.9 min, positive)
R
in the case of 1, 2, 4, and 5, and D-glucose and L-rhamnose
(tR 4.2 min, negative) in the case of 3 by comparison with
an authentic sample.
1
23