1928
H. Kato et al. / Phytochemistry 71 (2010) 1925–1929
(4 mg), 4 (30 mg), 5 (4 mg), 6 (9 mg), 7 (22 mg), 8 (64 mg), 9
(5 mg), 10 (8 mg), 11 (10 mg), and 12 (7 mg).
5,7-Dihydroxy-2-propylchromone
: þ0:25 (c 0.1, MeOH). IR (KBr): mmax 3447, 1647, 1560, 1508,
1499, 1474, 1459, 1420, 1357, 1167 cmꢀ1 1H NMR (CD3OD,
(3a):
colorless
solid.
½ ꢁ
a 2D3
.
3.4. (ꢀ)-Aromadendrin 3-O-b-
D
-glucopyranoside (1)
500 MHz): d 6.31 (1H, d, J = 2.3 Hz, H-8), 6.18 (1H, d, J = 2.3 Hz,
H-6), 6.05 (1H, s, H-3), 2.61 (2H, t, J = 7.4 Hz, H-10), 1.76 (2H, qt,
J = 7.4 Hz, H-20), 1.02 (3H, t, J = 7.4 Hz, H-30). 13C NMR (CD3OD,
125 MHz): d 184.1 (qC, C-4), 172.4 (qC, C-2), 166.2 (qC, C-7),
163.3 (qC, C-5), 160.0 (qC, C-9), 108.5 (CH, C-3), 105.4 (qC, C-10),
100.1 (CH, C-6), 95.0 (CH, C-8), 36.9 (CH2, C-10), 21.3 (CH2, C-20),
13.8 (CH3, C-30). ESI–MS (positive) m/z: 221 [M + H]+.
Pale yellow solid. ½a D25
ꢁ
: ꢀ84:1 (c 0.7, MeOH). UV (MeOH): kmax
nm (loge) 213 (4.36), 225 (sh) (4.31), 291 (4.11), 330 (sh) (3.66).
IR (KBr): mmax 3423, 1638, 1543, 1522, 1459, 1384, 1260, 1170,
1082, 1026 cmꢀ1. For 1H NMR (CD3OD, 500 MHz) and 13C NMR
(CD3OD, 125 MHz) spectroscopic data, see Table 1. ESI–MS (posi-
tive) m/z: 473 [M + Na]+. HR–ESI–MS (positive) m/z: Found:
473.1056; Calc. for [C21H22O11Na]+: 473.1060. CD (MeOH): [h]25
(nm) ꢀ11759 (323), +13335 (293), +7574 (229), ꢀ18728 (216).
Agrimonolide (4a): colorless solid. ½a D24
: ꢀ7:4 (c 0.3, MeOH). UV
ꢁ
(MeOH): kmax nm (loge) 217 (4.30), 268 (4.00), 300 (3.66). IR (KBr):
mmax 3425, 1656, 1631, 1510, 1383, 1245, 1168, 1117 cmꢀ1 1H
.
NMR (CD3OD, 500 MHz): d 7.15 (2H, d, J = 8.8 Hz, H-20 and 60),
6.83 (2H, d, J = 8.7 Hz, H-30 and 50), 6.21 (1H, d, J = 2.3 Hz, H-7),
6.20 (1H, d, J = 2.0 Hz, H-5), 4.47 (1H, m, H-3), 3.75 (3H, s, OCH3),
2.88 (2H, m, H2-4), 2.82 (1H, ddd, J = 14.4, 9.6, 5.3 Hz, H-200a),
2.73 (1H, ddd, J = 13.8, 9.2, 7.1 Hz, H-200b), 2.07(1H, m, H-100a),
1.97(1H, m, H-100b). 13C NMR (CD3OD, 125 MHz): d 171.7 (qC, C-
1), 166.4 (qC, C-8), 165.7 (qC, C-6), 159.6 (qC, C-40), 143.5 (qC, C-
10), 134.5 (qC, C-10), 130.4 (CH, C-20 and 60), 115.0 (CH, C-30 and
50), 108.0 (CH, C-5), 102.3 (qC, C-7), 101.6 (CH, C-9), 79.9 (CH, C-
3), 55.7 (CH3, OCH3), 37.9 (CH2, C-100), 34.0 (CH2, C-4), 31.2 (CH2,
C-200). ESI–MS (positive) m/z: 337 [M + Na]+. CD (MeOH): [h]25
(nm) ꢀ3037 (301), +6789 (269), ꢀ4511 (231).
3.5. Desmethylagrimonolide 6-O-b-
D
-glucopyranoside (2)
Pale yellow solid. ½a D20
ꢁ
: ꢀ12:7 (c 0.3, MeOH). UV (MeOH): kmax
nm (loge) 216 (4.21), 263 (3.85), 303 (3.45). IR (KBr): mmax 3445,
1637, 1518, 1459, 1384, 1255, 1205, 1175, 1078 cmꢀ1. For 1H
NMR (CD3OD, 500 MHz) and 13C NMR (CD3OD, 125 MHz) spectro-
scopic data, see Table 2. ESI–MS (positive) m/z: 485 [M + Na]+. HR–
ESI–MS (positive) m/z: Found: 485.1427; Calc. for [C23H26O10Na]+:
485.1424. CD (MeOH): [h]25 (nm) ꢀ3021 (302), +3849 (267),
ꢀ4205 (227).
3.6. 5,7-Dihydroxy-2-propylchromone 7-O-b-
D
-glucopyranoside (3)
3.8. Methylation of 2 and 4
Colorless solid. ½a D20
ꢁ
: ꢀ63:9 (c 0.4, MeOH). UV (MeOH): kmax nm
(log
e
) 229 (4.18), 248 (4.19), 285 (3.75), 311 (sh) (3.57). IR (KBr):
Compounds 2 and 4 (each 0.5 mg) were individually dissolved
in MeOH (0.1 ml), respectively. After TMSCHN2 (0.6 M in ether,
20 ll) was added, the solutions stood for 6 h at room temperature.
The residues were obtained by evaporation in vacuo and purified
by preparative TLC with CHCl3–MeOH–H2O (60:15:1) to give a
same compound 2b (each 0.5 mg). The structure of 2b was deter-
mmax 3423, 1656, 1560, 1543, 1509, 1459, 1266, 1176, 1077 cmꢀ1
.
For 1H NMR (CD3OD, 500 MHz) and 13C NMR (CD3OD, 125 MHz)
spectroscopic data, see Table 3. ESI–MS (positive) m/z: 405
[M + Na]+. HR–ESI–MS (positive) m/z: Found: 405.1164; Calc. for
[C18H22O9Na]+: 405.1161.
mined as methylagrimonolide 6-O-b-
D
-glucopyranoside.
-glucopyranoside (2b): colorless
: ꢀ11:2 (c 0.1, MeOH). UV (MeOH): kmax nm (log ) 216
3.7. Enzymatic hydrolysis of 1–4
Methylagrimonolide 6-O-b-D
solid. ½a 2D2
ꢁ
e
Compound 1 (1.0 mg) was dissolved in 0.1 M NaoAc acetate
buffer (pH 4.0, 1.0 ml), then naringinase (1.0 U, Sigma Chemical
Co.) was added. After the reaction solution was stirred at 40 °C
for 4 h, it was passed through a Sep-pak C18 cartridge, then eluted
with H2O (10 ml). Further elution by MeOH (10 ml) afforded the
aglycone 1a (0.6 mg), which was identified as (ꢀ)-aromadendrin
by comparison with literature data (Takahashi et al., 1988; Prescott
et al., 2002).
(4.45), 259 (4.05), 298 (3.70). IR (KBr): mmax 3397, 1708, 1610,
1514, 1252, 1102, 1082, 1057, 1038 cmꢀ1 1H NMR (CD3OD, 500
.
MHz): d 7.13 (2H, d, J = 8.8 Hz, H-20 and 60), 6.83 (2H, d, J =
8.7 Hz, H-30 and 50), 6.74 (1H, d, J = 2.0 Hz, H-7), 6.60 (1H, d,
J = 2.0 Hz, H-5), 5.00 (1H, d, J = 7.5 Hz, Glc-H-1), 4.34 (1H, m, H-
3), 3.92 (1H, dd, J = 12.1, 2.3 Hz, Glc-H-6a), 3.88 (3H, s, 80-OCH3),
3.75 (3H, s, 40-OCH3), 3.66 (1H, dd, J = 12.1, 6.4 Hz, Glc-H-6b),
3.48 (3H, m, Glc-H-2, 3 and 5), 3.34 (1H, m, Glc-H-4), 2.93 (1H,
dd, J = 16.7, 4.3 Hz, H-4a), 2.88 (1H, dd, J = 16.1, 10.1 Hz, H-4b),
2.81 (1H, ddd, J = 14.5, 9.2, 5.3 Hz, H-200a), 2.72 (1H, ddd, J = 13.9,
9.1, 7.1 Hz, H-200b), 2.05 (1H, m, H-100a), 1.95 (1H, m, H-100b). 13C
NMR (CD3OD, 500 MHz): d 165.5 (qC, C-1), 164.5 (qC, C-8), 164.3
(qC, C-6), 159.6 (qC, C-40), 145.7 (qC, C-10), 134.5 (qC, C-10),
130.4 (CH, C-20 and 60), 115.0 (CH, C-30 and 50), 108.4 (qC, C-9),
108.3 (CH, C-5), 101.8 (CH, Glc-C-1), 101.0 (CH, C-7), 78.7 (CH,
Glc-C-5), 78.5 (CH, C-3), 78.0 (CH, Glc-C-3), 74.8 (CH, Glc-C-2),
71.6 (CH, Glc-C-4), 62.7 (CH2, Glc-C-6), 56.6 (CH3, 80-OCH3), 55.7
(CH3, 40-OCH3), 37.7 (CH2, C-100), 35.4 (CH2, C-4), 31.2 (CH2, C-200).
ESI–MS (positive) m/z: 513 [M + Na]+.
Enzymatic hydrolysis of 2 (1.0 mg), 3 (1.0 mg), and 4 (3.0 mg)
were carried out by the same procedure as 1 to afford the agly-
cones 2a (desmethylagrimonolide, 0.5 mg) (Yamato, 1958), 3a
(5,7-dihydroxy-2-propylchromone, 0.8 mg) (Alves et al., 1999)
and 4a (agrimonolide, 2.5 mg) (Arakawa et al., 1968; Yamato and
Hashigaki, 1976).
Desmethylagrimonolide (2a): pale yellow solid. ½a D23
: ꢀ3:5 (c
ꢁ
0.1, MeOH). UV (MeOH): kmax nm (loge) 215 (3.87), 269 (3.54),
302 (3.26). IR (KBr): mmax 3445, 1637, 1518, 1459, 1384, 1255,
1205, 1175, 1078 cmꢀ1 1H NMR (CD3OD, 500 MHz): d 7.04 (2H,
.
d, J = 8.5 Hz, H-20 and 60), 6.71 (2H, d, J = 8.7 Hz, H-30 and 50), 6.18
(1H, d, J = 2.3 Hz, H-7), 6.16 (1H, d, J = 2.3 Hz, H-5), 4.47 (1H, m,
H-3), 2.87 (2H, m, H2-4), 2.79 (1H, ddd, J = 14.7, 9.4, 5.3 Hz, H-
200a), 2.69 (1H, ddd, J = 14.0, 9.2, 7.1 Hz, H-200b), 2.06 (1H, m, H-
100a), 1.96 (1H, m, H-100b). 13C NMR (CD3OD, 125 MHz): d 171.8
(qC, C-1), 166.8 (qC, C-8), 165.7 (qC, C-6), 156.7 (qC, C-40), 143.4
(qC, C-10), 133.3 (qC, C-10), 130.4 (CH, C-20 and 60), 116.3 (CH, C-
30 and 50), 108.6 (CH, C-5), 102.5 (qC, C-9, CH, C-7), 79.9 (CH, C-
3), 38.0 (CH2, C-100), 34.0 (CH2, C-4), 31.2 (CH2, C-200). ESI–MS (neg-
ative) m/z: 299 [M–H]ꢀ. CD (MeOH): [h]25 (nm) ꢀ1133 (302),
+2137 (269), ꢀ1500 (230).
3.9. Acid hydrolysis and determination of the absolute configuration of
sugars in 1–4
Acid hydrolysis of compound 1–4 (each 0.5 mg) was carried out
by the same procedure as in our previous study (Li et al., 2009).
Briefly, acid hydrolysis was carried out in 1 M HCl (dioxane-H2O,
1:1, 200 ll) at 100 °C for 1 h under Ar. After the solution was ex-
tracted with EtOAc (1 ml ꢂ 3) to remove the aglycone, the aqueous
layer was evaporated to give the sugar fraction. The solution of the