634
Vol. 57, No. 6
Table 1. 13C-NMR Spectral Data for 1—3 (in Pyridine-d5, 125 MHz)
Experimental
All instruments and materials used were the same as those cited in a pre-
vious report,15) unless otherwise specified.
1
2
3
1
2
3
Plant Material The fruits of S. viarum were collected in the Medical
Plant Garden of Kumamoto University, Kumamoto Prefecture, Japan, in De-
cember 2003, and identified by Professor Toshihiro Nohara, Faculty of Phar-
maceutical Sciences, Sojo University.
Ag-1
2
37.6
30.2
78.2
39.0
140.8
122.0
32.3
32.2
50.5
37.0
21.4
40.2
42.4
57.0
24.6
28.2
53.1
12.0
19.4
41.5
12.5
73.0
33.8
31.2
34.4
75.3
17.6
37.5
30.2
78.2
39.0
140.9
121.7
32.2
31.4
50.1
37.2
20.8
38.9
41.3
55.5
35.3
119.1
72.4
13.8
19.4
34.2
17.3
87.5
30.0
32.0
34.2
75.1
17.7
50.4
37.4
30.0
77.5
38.7
140.8
121.6
32.2
31.5
50.2
37.0
21.0
39.8
40.4
56.3
32.1
80.9
62.3
16.1
19.3
38.4
15.0
120.1
33.1
33.8
83.8
77.3
24.3
Glc-1 100.3
100.4
77.9
78.5
78.9
76.9
61.4
2
3
4
5
77.9
78.5
78.8
76.9
61.4
3
4
5
6
7
8
9
Extraction and Isolation The fresh fruits of S. viarum (3.05 kg) were
extracted with MeOH at room temperature, and the solvent was removed
under reduced pressure to give a syrup (192.8 g). The MeOH extract was
chromatographed over Diaion HP20 column (H2O, MeOH, 50% aceone,
acetone) to afford fractions (frs.) 1—4. Fraction 2 (38.0 g) was subjected to
silica gel column chromatography (CC) [Merck. Art. 7734, CHCl3–MeOH–
H2O (14 : 2 : 0.1, 10 : 2 : 0.1, 8 : 2 : 0.2, 6 : 4 : 1, 0 : 1 : 0)] to afford frs. 2.1—
2.7. Chromatography of fr. 2.4 over Chromatorex ODS column (60%
MeOH, 70% MeOH, 80% MeOH, 90% MeOH, MeOH) produced frs.
2.4.1—2.4.5 and 5 (6569 mg). Fraction 2.4.2 (1735 mg) was successively
subjected to Chromatorex ODS CC (60% MeOH, 70% MeOH, 80% MeOH,
90% MeOH, MeOH) and HPLC (column, COSMOSIL 5C18 AR-II, Nacalai
Tesque, Inc., 20 mmꢃ250 mm; solvent, 80% MeOH) to afford fr. 2.4.2.1
(16 mg), 3 (8 mg) and 5 (14 mg). Fraction 2.4.4 (1239 mg) was chromato-
graphed over Chromatorex ODS column (60% MeOH, 70% MeOH, 80%
MeOH, 90% MeOH, MeOH) to produce frs. 2.4.4.1—2.4.4.5 and 5 (69 mg).
Fractions 2.4.4.1 (47 mg), 2.4.4.3 (107 mg), and 2.4.4.4 (494 mg) were each
subjected to HPLC (70% MeOH) under the conditions similar to those used
for fr. 2.4.2 to afford 2 (14 mg) from fr. 2.4.4.1, fr. 2.4.4.3.1 from fr. 2.4.4.3,
and 10 (18 mg) and 8 (19 mg) from fr. 2.4.4.4. Chromatography of fr.
2.4.4.3.1 (65 mg) over silica gel column [Merck. Art. 9385, CHCl3–
MeOH–H2O (10 : 2 : 0.1, 8 : 2 : 0.2, 0 : 1 : 0)] produced 4 (25 mg). Fraction
2.5 (15.6 g) was subjected to Chromatorex ODS CC (65% MeOH, 70%
MeOH, 80% MeOH, 90% MeOH, MeOH) to afford frs. 2.5.1—2.5.5. Frac-
tion 2.5.1 (11.04 g) was successively subjected to Chromatorex ODS CC
(50% MeOH, 60% MeOH, 70% MeOH), silica gel CC [Merck. Art. 9385,
CHCl3–MeOH–H2O (10 : 2 : 0.1, 8 : 2 : 0.2, 7 : 3 : 0.5, 6 : 4 : 1, 0 : 1 : 0)], and
HPLC under the same conditions as those used for fr. 2.4.4.1 to afford
fr. 2.5.1.1 and fr. 2.5.1.2 (25 mg). Fraction 2.5.1.1 (270 mg) was chro-
matographed over Chromatorex ODS column (60% MeOH, 70% MeOH,
80% MeOH, 90% MeOH, MeOH) to produce frs. 2.5.1.1.1—2.5.1.1.3.
HPLC (fr. 2.5.1.1.1, 80% MeOH; fr. 2.5.1.1.3, 75% MeOH) of fr. 2.5.1.1.1
(127 mg) and fr. 2.5.1.1.3 (124 mg) under the conditions similar to those
used for fr. 2.4.4.1 yielded 6 (43 mg) from fr. 2.5.1.1.1 and 9 (45 mg) from
fr. 2.5.1.1.3. Fraction 2.5.2 (3.1 g) was subjected to silica gel CC [Merck.
Art. 9385, CHCl3–MeOH–H2O (14 : 2 : 0.1, 10 : 2 : 0.1, 8 : 2 : 0.2, 7 : 3 : 0.5,
6 : 4 : 1, 0 : 1 : 0)] to afford frs. 2.5.2.1—2.5.2.6. Fractions 2.5.2.3 (150 mg)
and 2.5.2.5 (1945 mg) were each successively subjected to Chromatorex
ODS CC (50% MeOH, 60% MeOH, 65% MeOH, 70% MeOH, 80% MeOH,
90% MeOH, MeOH) and HPLC (fr. 2.5.2.3, 75% MeOH; fr. 2.5.2.5, 65%
MeOH) under the conditions similar to those used for fr. 2.4.4.1 to produce
6
Gal-1
100.4
76.2
84.9
70.1
74.9
62.1
3
3
4
5
6
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
OCH3
Rha-1 102.0
102.0 102.0
2
3
4
5
6
72.5
72.8
74.1
69.5
18.6
72.5
72.8
74.2
69.5
18.6
103.0
72.7
72.5
73.9
70.5
18.5
72.3
72.7
74.0
69.3
18.5
Rhaꢄ-1 102.9
2
3
4
5
72.7
72.5
73.9
70.4
18.5
6
Xyl-1
106.5
74.5
78.1
70.8
66.9
105.2
75.2
78.3
71.5
78.2
62.6
2
3
4
5
Glcꢄ-1 104.9
104.9
75.3
78.7
71.8
78.0
62.9
2
3
4
5
6
75.2
78.6
71.8
77.8
62.9
d in ppm from tetramethylsilane (TMS). Glc, glucopyranosyl; Gal, galactopyranosyl;
Rha, rhamnopyranosyl; Xyl, xylopyranosyl; Ag, aglycone moiety.
2: Amorphous powder. [a]D31 ꢂ70.3° (cꢁ0.9, pyridine). Positive FAB-MS
m/z: 1085 [MꢀNa]ꢀ. HR-positive FAB-MS m/z: 1085.5560 [MꢀNa]ꢀ
(Calcd for C52H86O22Na: 1085.5508). 1H-NMR (in pyridine-d5, 500 MHz) d:
6.37 (1H, s, H-1 of Rha), 5.83 (1H, s, H-1 of Rhaꢄ), 5.30 (1H, br d,
5 (106 mg) from fr. 2.5.2.3 and 7 (94 mg) from fr. 2.5.2.5. Fraction 2.5.3 Jꢁ4.5 Hz, H-6 of Ag), 4.86 (1H, d, Jꢁ8.1 Hz, H-1 of Glcꢄ), 4.81 (1H, d,
(1.6 g) was successively subjected to silica gel CC [Merck. Art. 9385, Jꢁ3.5 Hz, H-2 of Rha), 4.67 (1H, Jꢁ3.5 Hz, H-2 of Rhaꢄ), 4.61 (1H, dd,
CHCl3–MeOH–H2O (10 : 2 : 0.1, 8 : 2 : 0.2, 7 : 3 : 0.5, 6 : 4 : 1, 0 : 1 : 0)] and
HPLC under the same conditions as those used for fr. 2.5.2.3 to afford 6
Jꢁ3.5, 9.0 Hz, H-3 of Rha), 4.55 (1H, dd, Jꢁ2.5, 12.0 Hz, H-6 of Glcꢄ),
4.52 (1H, dd, Jꢁ3.5, 9.0 Hz, H-3 of Rhaꢄ), 4.08 (1H, dd, Jꢁ3.5, 12.0 Hz, H-
(84 mg). Fraction 2.5.5 (3.0 g) was successively subjected to silica gel CC 6 of Glc), 4.04 (1H, dd, Jꢁ8.0, 9.0 Hz, H-2 of Glcꢄ), 3.98 (1H, dd, Jꢁ7.5,
[Merck. Art. 9385, CHCl3–MeOH–H2O (14 : 2 : 0.1, 10 : 2 : 0.1, 8 : 2 : 0.2,
6 : 4 : 1, 0 : 1 : 0)], Chromatorex ODS CC (60% MeOH, 70% MeOH, 80%
MeOH, 90% MeOH, MeOH), and HPLC under the same conditions as those
used for fr. 2.4.4.1 to afford 1 (29 mg), 10 (18 mg), and 3 (72 mg).
9.5 Hz, Ha-26 of Ag), 3.86 (1H, m, H-3 of Ag), 3.68 (1H, dd, Jꢁ5.5, 9.0 Hz,
Hb-26 of Ag), 3.36 (3H, s, OCH3), 2.78 (1H, br dd, Jꢁ3.5, 13.5 Hz, Ha-4 of
Ag), 2.71 (1H, br dd, Jꢁ13.5, 13.5 Hz, Hb-4 of Ag), 2.25 (1H, m, H-20 of
Ag), 2.12 (1H, dd, Jꢁ6.5, 12.0 Hz, H-15 of Ag), 1.96 (1H, d, Jꢁ3.0 Hz, H-
1: Amorphous powder. [a]D31 ꢂ90.3° (cꢁ3.3, pyridine). Positive FAB-MS 17 of Ag), 1.75 (3H, d, Jꢁ6.5 Hz, H3-6 of Rha), 1.62 (3H, d, Jꢁ6.5 Hz, H3-6
m/z: 1057 [MꢀNa]ꢀ. HR-positive FAB-MS m/z: 1057.5630 [MꢀNa]ꢀ of Rhaꢄ), 1.08 (3H, d, Jꢁ7.5 Hz, H3-21 of Ag), 1.05 (3H, d, Jꢁ7.0 Hz, H3-27
(Calcd for C51H86O21Na: 1057.5559). 1H-NMR (in pyridine-d5, 500 MHz) d: of Ag), 1.05 (3H, s, H3-19 of Ag), 0.84 (3H, s, H3-18 of Ag). 13C-NMR data:
6.38 (1H, s, H-1 of Rha), 5.85 (1H, s, H-1 of Rhaꢄ), 5.36 (1H, d, Jꢁ4.5 Hz,
H-6 of Ag), 4.85 (1H, d, Jꢁ8.0 Hz, H-1 of Glcꢄ), 4.68 (1H, br s, H-2 of
Rha), 4.68 (1H, s, H-2 of Rhaꢄ), 4.96 (1H, d, Jꢁ7.5 Hz, H-1 of Glc), 4.85
(1H, d, Jꢁ7.5 Hz, H-1 of Glcꢄ), 4.85 (1H, d, Jꢁ3.5 Hz, H-2 of Rha), 4.70
(1H, d, Jꢁ3.0 Hz, H-2 of Rhaꢄ), 4.64 (1H, dd, Jꢁ3.5, 9.5 Hz, H-3 of Rha),
see Table 1.
3: Amorphous powder. [a]D31 ꢂ73.2° (cꢁ8.0, pyridine). Positive FAB-MS
m/z: 1055 [MꢀNa]ꢀ. HR-positive FAB-MS m/z: 1055.5056 [MꢀNa]ꢀ
(Calcd for C50H80O22Na: 1055.5039). 1H-NMR (in pyridine-d5, 500 MHz) d:
6.22 (1H, s, H-1 of Rha), 5.32 (1H, br s, H-6 of Ag), 5.01 (1H, d, Jꢁ7.5 Hz,
4.58 (1H, dd, Jꢁ2.5, 12.0 Hz, Ha-6 of Glcꢄ), 4.56 (1H, dd, Jꢁ3.0, 9.5 Hz, H- H-1 of Xyl), 4.96 (1H, d, Jꢁ7.5 Hz, H-1 of Gal), 4.94 (1H, d, Jꢁ7.5Hz, H-1
3 of Rhaꢄ), 4.35 (1H, dd, Jꢁ9.5, 9.5 Hz, H-4 of Rhaꢄ), 4.28 (1H, dd, Jꢁ8.5, of Glcꢄ), 4.90 (1H, dq, Jꢁ9.0, 6.0 Hz, H-5 of Rha), 4.86 (1H, br d,
8.5 Hz, H-3 of Glcꢄ), 4.10 (1H, dd, Jꢁ3.0,12.0 Hz, Hb-6 of Glc), 4.06 (1H,
dd, Jꢁ8.0, 9.0 Hz, H-3 of Glcꢄ), 3.97 (1H, dd, Jꢁ6.5, 9.0 Hz, Ha-26 of Ag),
Jꢁ3.0 Hz, H-2 of Rha), 4.76 (1H, br d, Jꢁ2.0 Hz, H-4 of Gal), 4.57 (1H, dd,
Jꢁ3.0, 9.0 Hz, H-3 of Rha), 4.52 (1H, dd, Jꢁ2.0, 11.5 Hz, Ha-6 of Glcꢄ),
3.65 (1H, dd, Jꢁ5.5, 9.0 Hz, Hb-26 of Ag), 2.82 (1H, br dd, Jꢁ3.0, 13.0 Hz, 4.02 (1H, dd, Jꢁ8.0, 8.5 Hz, H-2 of Glcꢄ), 3.89 (1H, d, Jꢁ9.5 Hz, Hb-27 of
Ha-4 of Ag), 2.74 (1H, dd, Jꢁ13.0, 13.0 Hz, Hb-4 of Ag), 1.78 (3H, d, Ag), 3.62 (1H, dd, Jꢁ10.5, 10.5 Hz, Hb-5 of Xyl), 2.80 (1H, br dd, Jꢁ4.0,
Jꢁ6.0 Hz, H3-6 of Rha), 1.63 (3H, d, Jꢁ6.0 Hz, H3-6 of Rhaꢄ), 1.19 (3H, d, 12.5 Hz, Ha-4 of Ag), 2.73 (1H, br dd, Jꢁ12.5, 12.5 Hz, Hb-4 of Ag), 1.67
Jꢁ6.5 Hz, H3-21 of Ag), 1.08 (3H, s, H3-19 of Ag), 1.05 (3H, d, Jꢁ6.5 Hz,
(3H, d, Jꢁ6.0 Hz, H3-6 of Rha), 1.40 (3H, s, H3-27 of Ag), 1.08 (3H, d,
Jꢁ6.5 Hz, H3-21 of Ag), 1.05 (3H, s, H3-19 of Ag), 0.80 (3H, s, H3-18 of
H3-27 of Ag), 0.73 (3H, s, H3-18 of Ag). 13C-NMR data: see Table 1.