388
J Nat Med (2011) 65:385–390
TFA (20:80)–(30:70)] to give compounds 11 (6.1 mg),
13 (1.3 mg), 14 (1.3 mg), 15 (1.7 mg), 20 (1.8 mg),
21 (1.8 mg), and 22 (1.6 mg). Fraction 3E (531 mg) was
subjected to preparative HPLC [column, Capsell-Pak Ph;
solvent, methanol–0.2% TFA (30:70)] to yield 30 (9.0 mg).
The water eluate, 10% MeOH eluate, 45% MeOH eluate,
90% MeOH eluate, and fractions 2A, 2B, 3A, 3B were
suspended in water (1.0 L), and partitioned with ethyl
acetate (1.0 L). The ethyl acetate layer (5.0 g) was sub-
jected to preparative LPLC and HPLC [columns, Ultra
Pack ODS-SM-50C-M; solvent, methanol–0.2% TFA
(35:65), ODS-100V; solvent, methanol–0.2% TFA (45:55)
and Mightysil RP-18 GP; solvent, acetonitrile–0.2% TFA
(20:80)] to yield compounds 12 (30.6 mg), 16 (16.4 mg), and
17 (1.5 mg). Thewaterlayer was passed througha DiaionHP-20
column (eluted with water; 5, 20, 50% MeOH, and MeOH). The
50% MeOH eluate (2.70 g) was subjected to preparative LPLC
and HPLC [columns, Ultra Pack ODS-SM-50C-M, AR-II; sol-
vent, acetonitrile–0.2% TFA (15:85); solvent, methanol–0.2%
TFA (35:65), ODS-AM; solvent, acetonitrile–0.2% TFA (15:85)
or (20:80)] to yield compounds 1 (2.6 mg), 18 (2.4 mg), and
28 (5.1 mg). A mixture of the ether layers was concentrated at
reduced pressure (25.8 g), suspended in 80% methanol (1.5 L),
and partitioned with hexane (1.0 L) three times. The 80%
methanol layer (3.27 g) was passed through an ODS (Cosmosil
140C18-OPN, 150 g) column and eluted with 40, 60, and 80%
MeOH. The 60% MeOH eluate (752 mg) was subjected to
preparative LPLC and HPLC [columns, Ultra Pack ODS-SM-
50C-M; solvent, methanol–0.2% TFA (60:40), ODS-100V;
solvent, acetonitril–0.2% TFA (40:60) and Mightysil RP-18 GP;
solvent, acetonitrile–0.2% TFA (22.5:77.5)–(35:65)] to yield
compounds 4 (2.1 mg), 5 (1.3 mg), 6 (2.6 mg), 19 (2.6 mg),
23 (1.3 mg), 24 (2.3 mg), 25 (3.9 mg), 27 (13.8 mg),
30 (8.6 mg), and 31 (2.3 mg).
11.0 Hz, H-500), 3.62 (1H, overlapped, H-500), 4.32 (1H, d,
J = 7.5 Hz, H-1000), 3.23 (1H, overlapped, H-2000),
3.20–3.35 (1H, overlapped, H-3000), 3.20–3.35 (1H, over-
lapped, H-4000), 3.20–3.35 (1H, overlapped, H-5000), 3.64
(1H, overlapped, H-6000), 3.84 (1H, overlapped, H-6000), 7.06
(1H, d, J = 1.5 Hz, H-20000), 6.79 (1H, d, J = 8.0 Hz,
H-50000), 6.96 (1H, dd, J = 8.0, 1.5 Hz, H-60000), 7.56 (1H, d,
J = 16.0 Hz, H-70000), 6.26 (1H, d, J = 16.0 Hz, H-80000).
13C-NMR (100 MHz, CD3OD) d: 131.5 (C-1), 117.2 (C-2),
146.1 (C-3), 144.7 (C-4), 116.4 (C-5), 121.3 (C-6), 36.5
(C-7), 72.3 (C-8), 103.9 (C-10), 74.9 (C-20), 85.2 (C-30),
70.9 (C-40), 74.7 (C-50), 69.4 (C-60), 106.9 (C-100), 75.1
(C-200), 77.6 (C-300), 71.0 (C-400), 67.3 (C-500), 104.7 (C-1000),
75.700(0C-2000), 77.8 (C-3000), 71.5 (C-4000), 77.9 (C-5000), 62.7
(C-60000), 127.8 (C-10000), 115.2 (C-20000), 146.9 (C-30000), 149.7
(C-4 ), 116.6 (C-50000), 123.0 (C-60000), 147.3 (C-70000), 115.2
(C-80000), 168.5 (C-90000). HMBC correlations: H-2/C-4, C-6,
C-7; H-5/C-1, C-3; H-6/C-2, C-4, C-7; H-7/C-1, C-2, C-6,
C-8; H-8/C-10; H-10/C-8; H-20/C-10; H-30/C-100; H-40/C-30,
C-5000, C-60, C-90000; H-60/C-1000; H-100/C-30; H-200/C-100; H-500/
C-1 ; H-1000/C-60; H-2000/C-1000; H-20000/C-40000, C-60000, C-70000;
H-50000/C-10000, C-30000, C-40000; H-60000/C-20000, C-40000, C-70000,
H-70000/C-10000, C-20000, C-60000, C-90000; H-80000/C-10000, C-90000.
Rashomoside B (2)
21
Colorless amorphous powder; ½aꢁD –50.08 (c 0.12, MeOH);
HRFABMS m/z 497.1276 (calcd. for C22H25O13, 497.1295).
UV kmax (MeOH) nm (log e): 281 (3.86), 206 (4.45). IR mmax
(KBr) cm-1: 3401, 1615, 1339, 1232, 1116, 1072. 1H-NMR
(400 MHz, CD3OD) d: 7.54 (1H, d, J = 3.0 Hz, H-2), 6.69
(1H, d, J = 9.0 Hz, H-5), 7.24 (1H, dd, J = 9.0, 3.0 Hz,
H-6), 4.79 (1H, overlapped, H-10), 3.45 (1H, overlapped,
H-20), 3.47 (1H, overlapped, H-30), 3.43 (1H, overlapped,
H-40), 3.73 (1H, m, H-50), 4.41 (1H, dd, J = 11.5, 7.0 Hz,
H-60), 4.69 (1H, dd, J = 11.5, 2.0 Hz, H-60), 7.29 (2H, s, H-200
and 600), 3.83 (6H, s, 300-OMe and 500-OMe). 13C-NMR
(100 MHz, CD3OD) d: 151.0 (C-1), 119.6 (C-2), 113.9 (C-3),
158.9 (C-4), 118.8 (C-5), 127.1 (C-6), 173.0 (C-7), 103.6
(C-10), 75.0 (C-20), 77.9 (C-30), 72.1 (C-40), 75.7 (C-50), 65.2
(C-60), 121.4 (C-100), 108.6 (C-200 and 600), 149.0 (C-300 and 500),
141.8 (C-400), 167.9 (C-700), 56.9 (C3-OMe and C5-OMe).
HMBC correlations: H-2/C-4, C-6, C-7; H-5/C-1, C-3, C-7;
H-6/C-4; H-10/C-1; H-40/C-30; H-50/C-10; H-60/C-700; H-200/
C-100, C-300, C-400, C-600, C-700; H-600/C-100, C-200, C-400, C-500, C-700;
H-OMe/C-300, C-500.
Rashomoside A (1)
21
Colorless amorphous powder; ½aꢁD –60.08 (c 0.05, MeOH);
HRFABMS m/z 795.2318 (calcd. for C34H44O20Na,
795.2322). UV kmax (MeOH) nm (log e): 330 (4.25), 291
(4.16), 203 (4.71). IR mmax (KBr) cm-1: 3421, 1629, 1264,
1
1160, 1120, 1045. H-NMR (400 MHz, CD3OD) d: 6.72
(1H, d, J = 1.5 Hz, H-2), 6.68 (1H, d, J = 8.0 Hz, H-5),
6.58 (1H, dd, J = 8.0, 1.5 Hz, H-6), 2.80 (2H, br t,
J = 7.5 Hz, H-7), 3.75 (1H, overlapped, H-8), 4.03 (1H,
m, H-8), 4.43 (1H, d, J = 7.5 Hz, H-10), 3.48 (1H, dd,
J = 7.5, 9.0 Hz, H-20), 3.83 (1H, overlapped, H-30), 5.00
(1H, dd, J = 9.5, 10.0 Hz, H-40), 3.75 (1H, overlapped,
H-50), 3.67 (1H, overlapped, H-60), 3.95 (1H, dd, J = 11.5,
1.5 Hz, H-60), 4.43 (1H, d, J = 7.5 Hz, H-100), 3.15 (1H,
dd, J = 8.5, 7.5 Hz, H-200), 3.35 (1H, overlapped, H-300),
3.35 (1H, overlapped, H-400), 3.07 (1H, dd, J = 10.0,
4,5-Di-O-(4-hydroxy-3,5-methoxy-benzoyl)-shikimic
acid (3)
Colorless amorphous powder; ½aꢁ2D3 –84.178 (c 0.48, MeOH);
HRFABMS m/z 557.1270 (calcd. for C25H26O13Na,
123