1278
Vol. 59, No. 10
ꢂ109.6 (cꢀ0.55, MeOH),1) kaempferol 3-O-glucoside (8), [a]D23 ꢂ13.9
(cꢀ0.11, MeOH),14) syringetin 3-O-glucoside (9), [a]D25 ꢂ11.9 (cꢀ0.16,
MeOH),15) arbutin (10), [a]D24 ꢂ45.6 (cꢀ0.62, MeOH),3) seguinoside A (11),
[a]D25 ꢂ72.9 (cꢀ0.68, MeOH),3) ardisiacrispin B (12), [a]D27 ꢂ16.7 (cꢀ1.56,
MeOH),9) and ardisicrenoside A (13), [a]D26 ꢂ17.2 (cꢀ1.69, MeOH).9)
Myrseguinoside A (1): Amorphous powder; [a]D27 ꢁ20.1 (cꢀ0.80,
MeOH); IR nmax (film) cmꢂ1: 3382, 2969, 2919, 1604, 1550, 1382, 1278,
1077, 1031, 920; 1H- and 13C-NMR (CD3OD): Table 1; HR-ESI-TOF-MS
(positive-ion mode) m/z: 355.1722 [MꢁNa]ꢁ (Calcd for C16H28O7Na:
355.1727).
HL-60 on IC50 value of 3.2ꢆ1.61 mM, while ardisicrenoside
A (13) was inactive (ꢇ100 mM).13) Considerating these re-
sults together with the weak activity of 4, the substituents
around the E-ring may be important for tumor cell growth
inhibitory activity.
Experimental
General Experimental Procedures Silica gel column chromatography
(CC) was performed on silica gel 60 (Merck, Darmstadt, Germany), and re-
versed-phase [octadecyl silica gel (ODS)] open CC (RPCC) on Cosmosil
75C18-OPN (Nacalai Tesque, Kyoto, Japan) (Fꢀ4 cm, Lꢀ20 cm). HPLC
was performed on an ODS column (Inertsil; GL Science, Tokyo, Japan;
Fꢀ10 mm, Lꢀ250 mm), and the eluate was monitored with a refractive
index monitor.
Myrseguinoside
B
(2): Amorphous powder; [a]D26 ꢂ38.6 (cꢀ0.08,
MeOH); IR nmax (film) cmꢂ1: 3393, 2961, 2851, 1650, 1603, 1506, 1361,
1201, 1087, 1025; UV lmax (MeOH) nm (log e): 358 (4.00), 252 (4.08), 211
(4.35); 1H- and 13C-NMR (DMSO-d6): Table 2; 1H-NMR (CD3OD) dH: 7.54
(1H, d, Jꢀ2 Hz, H-6ꢃ), 7.31 (1H, d, Jꢀ2 Hz, H-2ꢃ), 6.41 (1H, d, Jꢀ2 Hz, H-
8), 6.21 (1H, d, Jꢀ2 Hz, H-6), 5.22 (1H, d, Jꢀ7 Hz, H-1ꢄ), 3.94 (3H, s,
OMe), 3.90 (1H, dd, Jꢀ8, 7 Hz, H-2ꢄ), 3.83 (1H, dd, Jꢀ12, 3 Hz, H-5ꢄa),
3.81 (1H, ddd, Jꢀ3, 3, 1 Hz, H-4ꢄ), 3.64 (1H, dd, Jꢀ8, 3 Hz, H-3ꢄ), 3.45
(1H, dd, Jꢀ12, 1 Hz, H-5ꢄb); 13C-NMR (CD3OD) dC: 179.6 (C-4), 166.2 (C-
7), 163.2 (C-5), 158.7 (C-9), 158.5 (C-2), 149.3 (C-5ꢃ), 146.4 (C-3ꢃ), 139.0
(C-4ꢃ), 135.8 (C-3), 121.9 (C-1ꢃ), 111.1 (C-2ꢃ), 106.9 (C-6ꢃ), 105.8 (C-10),
104.5 (C-1ꢄ), 100.0 (C-6), 94.8 (C-8), 74.2 (C-3ꢄ), 73.0 (C-2ꢄ), 69.2 (C-4ꢄ),
67.2 (C-5ꢄ), 57.2 (OMe); HR-ESI-TOF-MS (positive-ion mode) m/z:
487.0843 [MꢁNa]ꢁ (Calcd for C21H20O12Na: 487.0846).
Optical rotations were measured on a JASCO P-1030 polarimeter. IR
spectra were measured on a Horiba FT-710 Fourier transform infrared spec-
trophotometer and UV spectra on a JASCO V-520 UV/Vis spectrophotome-
ter. NMR spectra were taken on a JEOL JNM-ECA 600 spectrometer at
1
600 MHz for H, and 150 MHz for 13C, respectively, with tetramethylsilane
(TMS) as an internal standard. Positive-ion HR-ESI-TOF-MS was recorded
on an Applied Biosystem QSTAR XL spectrometer. VERSA Max (Molecu-
lar Device, Silicon Valley, CA, U.S.A.) was used as a microplate reader.
Plant Material Fruits of M. seguinii were collected in Okinawa, Japan,
in July 1992, and a voucher specimen was deposited in the Herbarium of the
Department of Pharmacognosy, Graduate School of Biomedical Sciences,
Hiroshima University (No. 92-MS-Okinawa-0727).
Myrseguinoside
C
(3): Amorphous powder; [a]D27 ꢂ23.3 (cꢀ0.18,
MeOH); IR nmax (film) cmꢂ1: 3363, 2959, 2849, 1652, 1607, 1508, 1359,
1198, 1081, 1021; UV lmax (MeOH) nm (log e): 360 (3.84), 251 (3.88), 210
(4.17); 1H- and 13C-NMR (DMSO-d6): Table 2; 1H-NMR (CD3OD) dH: 7.55
(2H, s, H-2ꢃ, 6ꢃ), 6.37 (1H, d, Jꢀ2 Hz, H-8), 6.17 (1H, d, Jꢀ2 Hz, H-6), 5.21
(1H, d, Jꢀ7 Hz, H-1ꢄ), 3.94 (6H, s, OMe), 3.89 (1H, dd, Jꢀ9, 7 Hz, H-2ꢄ),
3.82 (1H, dd, Jꢀ12, 3 Hz, H-5ꢄa), 3.80 (1H, ddd, Jꢀ3, 3, 1 Hz, H-4ꢄ), 3.64
(1H, dd, Jꢀ9, 3 Hz, H-3ꢄ), 3.45 (1H, dd, Jꢀ12, 1 Hz, H-5ꢄb); 13C-NMR
(CD3OD) dC: 179.7 (C-4), 166.9 (C-7), 163.0 (C-5), 158.7 (C-9), 158.2 (C-
2), 149.1 (C-3ꢃ, 5ꢃ), 140.2 (C-4ꢃ), 135.7 (C-3), 122.0 (C-1ꢃ), 108.3 (C-2ꢃ,
6ꢃ), 105.1 (C-10), 104.6 (C-1ꢄ), 100.9 (C-6), 95.5 (C-8), 74.3 (C-3ꢄ), 73.1
(C-2ꢄ), 69.3 (C-4ꢄ), 67.3 (C-5ꢄ), 57.3 (OMe); HR-ESI-TOF-MS (positive-ion
mode) m/z: 501.1006 [MꢁNa]ꢁ (Calcd for C22H22O12Na: 501.1003).
Extraction and Isolation The air-dried fruits of M. seguinii (670 g)
were extracted with MeOH (4 l) three times by maceration. The MeOH ex-
tracts were combined and evaporated to dryness under vacuum to afford a
viscous gummy material (55.0 g). This residue (55.0 g) was applied to a sil-
ica gel column (Fꢀ5 cm, Lꢀ50 cm) using a stepwise gradient containing
increasing amounts of MeOH in CHCl3 [CHCl3 (3 l), CHCl3–MeOH (10 : 1,
3 l), (5 : 1, 3 l), (3 : 1, 3 l), and (2 : 1, 3 l), and MeOH (3 l)]. The residue
(2.12 g) of the CHCl3–MeOH (5 : 1) eluate obtained on silica gel CC was
subsequently subjected to RPCC with stepwise gradient elution with in-
creasing amounts of MeOH in H2O (10, 30, 50, 70, 90, 100% MeOH, 1 l).
The residue (300 mg) obtained from the 50% MeOH eluate was further ap-
plied to a Sephadex LH-20 column (Fꢀ2 cm, Lꢀ120 cm) with MeOH as an
eluate, 5-ml fractions (frs) being collected. The residue (264 mg in frs 1—
38) was further separated by HPLC (ODS) with 20% CH3CN to afford 1
(9.2 mg) from the peak at 9 min (flow rate: 3.0 ml/min). The residue
(19.4 mg in frs 39—58) was further separated by HPLC (ODS) with 20%
CH3CN to afford 9 (3.2 mg) and 8 (2.0 mg) from the peaks at 17 and 27 min,
respectively (flow rate: 3.0 ml/min). The residue (24.2 mg in frs 59—74) was
further separated by HPLC (ODS) with 25% CH3CN to afford 3 (12.8 mg)
from the peak at 12 min (flow rate: 3.0 ml/min).
The residue (2.57 g) of the CHCl3–MeOH (3 : 1) eluate obtained on silica
gel CC was subsequently subjected to RPCC with stepwise gradient elution
with increasing amounts of MeOH in H2O (10, 30, 50, 70, 90, 100% MeOH,
1 l). The residue (1.00 g) obtained from the 10% MeOH eluate was further
applied to a Sephadex LH-20 column (Fꢀ2 cm, Lꢀ120 cm) with MeOH as
an eluate, 5-ml frs being collected. The residue (510 mg in frs 25—45) was
further separated by HPLC (ODS) with 3% CH3CN to afford 10 (6.2 mg)
and 11 (6.8 mg) from the peaks at 11 min and 25 min, respectively (flow rate:
2.8 ml/min).
The residue (7.26 g) of the CHCl3–MeOH (2 : 1) eluate obtained on silica
gel CC was subsequently subjected to RPCC with stepwisw gradient elution
with increasing amounts of MeOH in H2O (10, 30, 50, 70, 90, 100% MeOH,
1 l). The residue (218 mg) obtained from the 50% MeOH eluate was further
applied to a Sephadex LH-20 column (Fꢀ2 cm, Lꢀ120 cm) with MeOH as
an eluate, 5-ml frs being collected. The residue (46.7 mg in frs 34—57) was
further separated by HPLC (ODS) with 25% CH3CN to afford 7 (8.5 mg), 2
(2.4 mg), and 6 (3.3 mg) from the peaks at 11, 14, and 17 min, respectively
(flow rate: 3.0 ml/min). The residue (217 mg) obtained from the 70% MeOH
eluate was further applied to a Sephadex LH-20 column (Fꢀ2 cm,
Lꢀ120 cm) with MeOH as an eluate, 5 ml fractions (frs) being collected.
The residue (102.2 mg in frs 9—19) was further separated by HPLC (ODS)
with 35% CH3CN to afford 4 (8.1 mg), 13 (16.9 mg), 12 (15.6 mg), and 5
(7.8 mg) from the peaks at 7, 9, 21, and 28 min, respectively (flow rate:
3.0 ml/min).
Myrseguinoside D (4): Amorphous powder; [a]D25 ꢂ12.9 (cꢀ0.81,
MeOH); IR nmax (film) cmꢂ1: 3395, 2928, 1753, 1649, 1511, 1457, 1261,
1073, 1039; 1H- and 13C-NMR (C5D5N): Table 3; 1H-NMR (CD3OD) dH:
5.32 (1H, dd, Jꢀ3, 3 Hz, H-12), 5.28 (1H, d, Jꢀ2 Hz, H-1ꢅ), 4.87 (1H, br d,
Jꢀ6 Hz, H-22), 4.62 (1H, d, Jꢀ8 Hz, H-1ꢄ), 4.60 (1H, d, Jꢀ8 Hz, H-1ꢄꢄ),
4.51 (1H, d, Jꢀ5 Hz, H-1ꢃ), 4.29 (1H, dd, Jꢀ4, 4 Hz, H-16), 4.11 (1H, dq,
Jꢀ9, 6 Hz, H-5ꢅ), 4.05 (1H, m, H-4ꢃ), 4.05 (1H, dd, Jꢀ13, 5 Hz, H-5ꢃa),
3.93 (1H, dd, Jꢀ3, 2 Hz, H-2ꢅ), 3.87 (1H, m, H-2ꢃ), 3.86 (1H, m, H-3ꢃ),
3.85 (1H, m, H-6ꢄꢄa), 3.82 (1H, dd, Jꢀ12, 2 Hz, H-6ꢄa), 3.75 (1H, dd, Jꢀ10,
3 Hz, H-3ꢅ), 3.66 (2H, m, H-6ꢄb, H-6ꢄꢄb), 3.55 (1H, m, H-5ꢃb), 3.44 (1H, m,
H-2ꢄ), 3.43 (1H, m, H-3ꢄ), 3.42 (1H, d, Jꢀ12 Hz, H-28a), 3.39 (1H, dd, Jꢀ9,
9 Hz, H-3ꢄꢄ), 3.38 (1H, dd, Jꢀ10, 9 Hz, H-4ꢅ), 3.28 (1H, m, H-4ꢄ, H-5ꢄꢄ),
3.27 (1H, m, H-4ꢄꢄ), 3.25 (1H, m, H-5ꢄ), 3.22 (1H, d, Jꢀ12 Hz, H-28b), 3.19
(1H, br d, Jꢀ11 Hz, H-21ax), 3.19 (1H, dd, Jꢀ9, 8 Hz, H-2ꢄꢄ), 3.15 (1H, dd,
Jꢀ12, 5 Hz, H-3), 2.15 (1H, dd, Jꢀ13, 13 Hz, H-19ax), 2.09 (1H, m, H-
15ax), 2.05 (1H, ddd, Jꢀ11, 6, 2 Hz, H-21 eq), 1.90 (2H, m, H2-11), 1.86
(1H, m, H-2 eq), 1.78 (1H, dd, Jꢀ13, 6 Hz, H-18), 1.74 (1H, m, H-2ax), 1.73
(1H, m, H-1eq), 1.65 (1H, m, H-9), 1.61 (1H, m, H-7ax), 1.60 (1H, m, H-
6eq), 1.51 (1H, ddd, Jꢀ13, 6, 2 Hz, H-19eq), 1.46 (1H, m, H-7eq), 1.45 (1H,
m, H-6ax), 1.44 (1H, m, H-15eq), 1.40 (3H, s, H-27), 1.30 (1H, d, Jꢀ6 Hz,
H-6ꢅ), 1.13 (3H, s, H-29), 1.06 (3H, s, H-23), 0.98 (1H, m, H-1ax), 0.98
(3H, s, H-25), 0.94 (3H, s, H-26), 0.85 (3H, s, H-24), 0.80 (1H, br d,
Jꢀ12 Hz, H-5); 13C-NMR (CD3OD) dC: 184.3 (C-30), 141.7 (C-13), 125.9
(C-12), 105.2 (C-1ꢃ), 104.7 (C-1ꢄ), 104.0 (C-1ꢄꢄ), 101.9 (C-1ꢅ), 90.9 (C-3),
81.4 (C-22), 79.7 (C-2ꢃ), 79.1 (C-2ꢄ), 78.9 (C-3ꢄ), 78.0 (C-3ꢄꢄ), 77.9 (C-5ꢄꢄ),
77.7 (C-5ꢄ), 76.6 (C-4ꢃ), 76.4 (C-2ꢄꢄ), 74.5 (C-4ꢅ), 72.8 (C-3ꢃ), 72.3 (C-2ꢅ),
72.1 (C-3ꢅ), 71.9 (C-4ꢄꢄ), 71.7 (C-4ꢄ), 70.4 (C-5ꢅ), 68.9 (C-16), 66.1 (C-28),
64.7 (C-5ꢃ), 62.9 (C-6ꢄꢄ), 62.8 (C-6ꢄ), 57.1 (C-5), 48.2 (C-9), 44.3 (C-20),
44.2 (C-19), 43.6 (C-18), 43.5 (C-14), 43.3 (C-17), 41.5 (C-21), 41.1 (C-8),
40.4 (C-4), 40.0 (C-1), 37.9 (C-10), 35.6 (C-15), 34.3 (C-7), 28.7 (C-23),
27.9 (C-27), 27.1 (C-2), 24.5 (C-11), 20.9 (C-29), 19.4 (C-6), 18.3 (C-6ꢅ),
18.1 (C-26), 17.0 (C-24), 16.3 (C-25); HR-ESI-TOF-MS (positive-ion
mode) m/z: 1111.5285 [MꢁNa]ꢁ (Calcd for C53H84O23Na: 1111.5295).
Myrseguinoside
E
(5): Amorphous powder; [a]D26 ꢂ14.2 (cꢀ0.50,
MeOH); IR nmax (film) cmꢂ1: 3399, 2927, 1649, 1509, 1455, 1260, 1073,
1044; 1H- and 13C-NMR (C5D5N): Table 3; 1H-NMR (CD3OD) dH: 5.28
(1H, d, Jꢀ2 Hz, H-1ꢅ), 4.61 (1H, d, Jꢀ8 Hz, H-1ꢄ), 4.60 (1H, d, Jꢀ8 Hz, H-
The known compounds (6—13) were identified by comparison of the
spectroscopic data with those reported in the literature, as follows.
Quercitrin (6), [a]D27 ꢂ100.2 (cꢀ0.22, MeOH),1) myricitrin (7), [a]D27