A R T I C L E S
Kanemitsu et al.
resin was washed with THF, MeOH, and DMF. The filtrate was
neutralized by Amberlite IR-120 (H+), filtered, and concentrated. The
residue was passed through an S-X1 gel permeation column with
toluene/ethyl acetate (3:1) as an eluent to separate the trisaccharide
(36 mg) and tetrasaccharide (30 mg) fractions. The trisaccharide fraction
was then purified by silica gel column chromatography using CH2Cl2/
MeOH/AcOH (40:1:0.5) as an eluent to provide 26a (12.6 mg) and
26b (23.4 mg). The tetrasaccharide fraction was also purified by silica
gel column chromatography using CH2Cl2/MeOH/H2O (15:5:0.8) as
an eluent to provide 25a (8.8 mg), 25b (3.6 mg), 25c (10.9 mg), and
25d (6.9 mg). Data for 25a: Rf 0.31 (CH2Cl2/MeOH/H2O, 15:5:0.8);
monohydrate (4:1, v/v; 5 mL) was stirred at 100 °C for 16 h. The
reaction mixture was concentrated in vacuo. The residue was treated
with pyridine (3 mL) and Ac2O (1.5 mL) at room temperature for 24
h. The reaction mixture was concentrated in vacuo. The residue was
then treated with 0.05 M NaOMe in MeOH (2 mL) at room temperature
for 3 h, neutralized with Amberlite IR-120 (H+), filtered, and
concentrated. The residue was purified by silica gel column chroma-
tography using CH2Cl2/MeOH/AcOH (40:1:0.5) as an eluent to provide
31b (20.0 mg 73%). Data for 28a: Rf 0.22 (CH2Cl2/MeOH/H2O, 8:2:
0.3); 1H NMR (CD3OD) δ 7.51-7.13 (m, 35 H), 5.20 (d, 1 H, J ) 3.1
Hz), 5.12 (d, 1 H, J ) 3.0 Hz), 5.01-4.28 (m, 22 H), 3.98-3.24 (m,
18 H), 2.83 (dd, 1 H, J ) 13.3, 4.9 Hz), 2.13 (t, 2 H, J ) 7.0 Hz), 2.10
(s, 3 H), 1.98 (s, 3 H), 1.81 (t, 1 H, J ) 13.3 Hz), 1.57-1.52 (m, 4 H),
1.34-1.25 (m, 2 H), 1.02 (d, 3 H, J ) 6.0); MALDI-TOF MS m/z
calcd for C86H104N2O25 (M) 1564, found 1587 (M + Na)+. Data for
28b: Rf 0.24 (CH2Cl2/MeOH/H2O, 8:2:0.3); 1H NMR (CD3OD) δ
7.58-7.11 (m, 35 H), 5.18 (d, 1 H, J ) 3.1 Hz), 5.00 (d, 1 H, J ) 3.0
Hz), 4.95-4.20 (m, 22 H), 3.98-3.28 (m, 18 H), 2.75 (dd, 1 H, J )
12.7, 4.8 Hz), 2.11 (t, 2 H, J ) 7.3 Hz), 2.10 (s, 3 H), 1.96 (s, 3 H),
1.79 (t, 1 H, J ) 12.7 Hz), 1.57-1.51 (m, 4 H), 1.38-1.26 (m, 2 H),
0.96 (d, 3 H, J ) 6.6); MALDI-TOF MS m/z calcd for C88H104N2O25
1564, found 1587 (M + Na)+. Data for 28c: Rf 0.21 (CH2Cl2/MeOH/
1
[R]D -18.9° (c 0.3, MeOH); H NMR (CD3OD) δ 7.97-7.09 (m, 39
H), 5.49 (d, 1 H, J ) 3.0 Hz), 5.05 (d, 1 H, J ) 3.3 Hz), 4.99-4.24
(m, 22 H), 4.07-3.29 (m, 18 H), 2.72 (dd, 1 H, J ) 13.5, 3.6 Hz),
2.20 (t, 2 H, J ) 7.4 Hz), 1.95 (s, 3 H), 1.85 (t, 1 H, J ) 13.5 Hz),
1.63-1.52 (m, 4 H), 1.42-1.31 (m, 2 H), 1.02 (d, 3 H, J ) 6.3);
MALDI-TOF MS m/z calcd for C92H106N2O27 1670, found 1693 (M +
Na)+. Data for 25b: Rf 0.38 (CH2Cl2/MeOH/H2O, 15:5:0.8); [R]D
1
-10.5° (c 0.2, MeOH); H NMR (CD3OD) δ 7.97-7.12 (m, 39 H),
5.49 (d, 1 H, J ) 3.0 Hz), 5.01 (d, 1 H, J ) 2.9 Hz), 4.88-4.25 (m,
22 H), 4.10-3.30 (m, 18 H), 2.90 (dd, 1 H, J ) 14.0, 3.0 Hz), 2.23 (t,
2 H, J ) 7.8 Hz), 1.96 (s, 3 H), 1.80 (t, 1 H, J ) 14.0 Hz), 1.65-1.50
(m, 4 H), 1.44-1.31 (m, 2 H), 0.93 (d, 3 H, J ) 5.9); MALDI-TOF
MS m/z calcd for C92H106N2O27 1670, found 1693 (M + Na)+; FAB
MS m/z calcd for C92H106N2O27 (M) 1670.7, found 1693.7 (M + Na)+.
Data for 25c: Rf 0.30 (CH2Cl2/MeOH/H2O, 15:5:0.8); [R]D -52.8° (c
0.4, MeOH); 1H NMR (CD3OD) δ 7.83-7.00 (m, 39 H), 5.57 (d, 1 H,
J ) 3.3 Hz), 5.18-4.29 (m, 23 H), 4.04-3.27 (m, 18 H), 2.90 (dd, 1
H, J ) 13.2, 5.0 Hz), 2.20 (t, 2 H, J ) 7.3 Hz), 2.02 (s, 3 H), 1.85 (t,
1 H, J ) 13.2 Hz), 1.59-1.52 (m, 4 H), 1.37-1.29 (m, 2 H), 0.99 (d,
3 H, J ) 6.3); MALDI-TOF MS m/z calcd for C92H106N2O27 1670,
found 1693 (M + Na)+. Data for 25d: Rf 0.34 (CH2Cl2/MeOH/H2O,
1
H2O, 8:2:0.3); H NMR (CD3OD) δ 7.53-7.11 (m, 35 H), 5.17 (d, 1
H, J ) 3.3 Hz), 5.14-4.28 (m, 23 H), 4.03-3.24 (m, 18 H), 2.91 (dd,
1 H, J ) 13.0, 4.8 Hz), 2.14 (t, 2 H, J ) 7.3 Hz), 2.02 (s, 3 H), 1.93
(s, 3 H), 1.85 (t, 1 H, J ) 13.0 Hz), 1.58-1.56 (m, 4 H), 1.34-1.28
(m, 2 H), 0.98 (d, 3 H, J ) 6.3); MALDI-TOF MS m/z calcd for
C86H104N2O25 (M) 1564, found 1587 (M + Na)+; FAB MS m/z calcd
for C86H104N2O25 (M) 1564.7, found 1585.6 (M + Na - 2H)-. Data
for 28d: Rf 0.22 (CH2Cl2/MeOH/H2O, 8:2:0.3); 1H NMR (CD3OD) δ
7.51-7.13 (m, 35 H), 5.02 (d, 1 H, J ) 3.0 Hz), 5.00-4.28 (m, 23 H),
4.00-3.24 (m, 18 H), 2.85 (dd, 1 H, J ) 13.3, 5.2 Hz), 2.13 (t, 2 H,
J ) 7.3 Hz), 2.03 (s, 3 H), 1.91 (s, 3 H), 1.82 (t, 1 H, J ) 13.3 Hz),
1.58-1.53 (m, 4 H), 1.34-1.25 (m, 2 H), 0.97 (d, 3 H, J ) 6.3);
MALDI-TOF MS m/z calcd for C86H104N2O25 (M) 1564, found 1587
(M + Na)+. Data for 31a: Rf 0.43 (CH2Cl2/MeOH/AcOH, 30:1:0.5);
1
15:5:0.8); [R]D -38.9° (c 0.3, MeOH); H NMR (CD3OD) δ 7.84-
7.00 (m, 39 H), 5.57 (d, 1 H, J ) 2.9 Hz), 5.08-4.23 (m, 23 H), 4.09-
3.27 (m, 18 H), 2.78 (dd, 1 H, J ) 13.2, 3.4 Hz), 2.19 (t, 2 H, J ) 7.3
Hz), 1.93 (s, 3 H), 1.83 (t, 1 H, J ) 13.2 Hz), 1.56-1.52 (m, 4 H),
1.40-1.28 (m, 2 H), 1.05 (d, 3 H, J ) 5.9); MALDI-TOF MS m/z
calcd for C92H106N2O27 1670, found 1693 (M + Na)+; HR-FAB MS
m/z calcd for C92H106N2O27 (M) 1670.6983, found 1708.6593 (M + K
- H)+. Data for 26a: Rf 0.15 (CH2Cl2/MeOH/AcOH, 40:1:0.5); [R]D
1
[R]D +12.0° (c 1.0, MeOH); H NMR (CD3OD) δ 7.38-7.22 (m, 35
H), 5.25 (d, 1 H, J ) 3.3 Hz), 5.20 (d, 1 H, J ) 3.6 Hz), 4.93-4.00
(m, 18 H), 3.89-3.29 (m, 16 H), 2.18 (t, 2 H, J ) 7.4 Hz), 1.68 (s, 3
H), 1.51 (m, 4 H), 1.28 (m, 2 H), 1.04 (d, 3 H, J ) 6.6); 13C NMR
(CD3OD) δ 180.3, 173.4, 141.0, 140.9, 140.8, 140.8, 140.6, 140.4,
140.3, 130.4, 130.3, 130.2, 130.2, 130.1, 130.0, 129.8, 129.7, 129.6,
129.5, 129.4, 102.5, 99.6, 98.4, 80.5, 80.2, 79.8, 79.0, 78.4, 77.5, 77.2,
77.1, 76.9, 76.0, 75.2, 75.2, 74.8, 74.5, 74.5, 72.5, 72.4, 70.9, 69.5,
53.6, 37.2, 31.1, 27.6, 27.2, 23.7, 17.8; HR-FAB MS m/z calcd for
C75H87NO17 (M) 1273.5974, found 1296.5929 (M + Na)+. Data for
31b: Rf 0.47 (CH2Cl2/MeOH/AcOH, 30:1:0.5); [R]D -72.8° (c 1.5,
MeOH); 1H NMR (CD3OD) δ 7.32-7.06 (m, 35 H), 5.10 (d, 1 H, J )
3.3 Hz), 4.86-4.19 (m, 18 H), 3.88-3.16 (m, 16 H), 2.11 (t, 2 H, J )
7.4 Hz), 1.83 (s, 3 H), 1.51-1.45 (m, 4 H), 1.27-1.14 (m, 2 H), 0.98
(d, 3 H, J ) 6.3); 13C NMR (CD3OD) δ 177.7, 173.2, 140.6, 140.4,
140.3, 140.3, 139.7, 139.6, 130.1, 130.0, 129.9, 129.8, 129.5, 129.5,
129.4, 129.3, 129.2, 129.1, 128.9, 128.8, 128.8, 128.7, 128.6, 128.5,
128.4, 128.3, 103.8, 102.9, 97.9, 81.8, 80.1, 79.9, 78.6, 77.1, 76.8, 76.4,
76.4, 76.3, 75.8, 75.3, 74.2, 74.0, 73.5, 73.1, 70.4, 69.4, 69.3, 67.5,
57.9, 35.1, 30.3, 26.6, 25.8, 23.5, 17.2; HR-FAB MS m/z calcd for
C75H87NO17 (M) 1273.5974, found 1296.5891 (M + Na)+.
Typical Procedure for the Debenzylation Reaction. A mixture of
compound 31b (36 mg, 0.028 mmol) and Pd(OH)2 (30 mg) in MeOH-
H2O (4:1, v/v; 5 mL) and AcOH (1 drop) was stirred under a H2
atmosphere for 48 h at room temperature. The reaction mixture was
filtered through a pad of Celite and the filtrate concentrated. The residue
was purified by silica gel column chromatography using CH2Cl2/MeOH/
H2O (5:4:1) as an eluent to provide 32b (17.0 mg 94%). Data for 29a:
Rf 0.47 (CH2Cl2/MeOH/H2O, 5:4:1); MALDI-TOF MS m/z calcd for
C37H62N2O25 934, found 957 (M + Na)+. Data for 29b: Rf 0.43 (CH2-
1
-20.1° (c 0.6, MeOH); H NMR (CD3OD) δ 7.79-7.13 (m, 39 H),
5.39 (d, 1 H, J ) 3.0 Hz), 5.17 (d, 1 H, J ) 3.3 Hz), 4.85-4.14 (m,
17 H), 4.04-3.29 (m, 16 H), 2.15 (t, 2 H, J ) 7.4 Hz), 1.51 (m, 4 H),
1.30 (m, 2 H), 1.05 (d, 3 H, J ) 6.3); 13C NMR (CD3OD) δ 177.5,
171.8, 170.1, 140.2, 140.1, 140.0, 139.7, 139.6, 139.5, 139.4, 139.1,
132.0, 129.8, 129.7, 129.6, 129.5, 129.5, 129.4, 129.4, 129.3, 129.3,
129.2, 129.1, 129.0, 129.0, 128.9, 128.9, 128.7, 128.7, 128.6, 128.6,
128.5, 128.2, 102.2, 98.3, 97.9, 79.6, 79.3, 79.1, 78.2, 77.1, 77.0, 76.7,
76.3, 76.2, 74.8, 74.3, 74.1, 74.0, 73.9, 73.4, 72.0, 71.4, 70.1, 70.0,
68.6, 54.0, 34.9, 30.3, 26.7, 25.8, 17.2; MALDI-TOF MS m/z calcd
for C81H89NO19 1379, found 1402 (M + Na)+; HR-FAB MS m/z calcd
for C81H89NO19 (M) 1379.6029, found 1402.5927 (M + Na)+.
Data for 26b: Rf 0.25 (CH2Cl2/MeOH/AcOH, 40:1:0.5); [R]D -68.9°
(c 1.0, MeOH); 1H NMR (CD3OD) δ 7.77-6.91 (m, 39 H), 5.51 (d, 1
H, J ) 3.3 Hz), 4.88-4.22 (m, 18 H), 4.02-3.14 (m, 16 H), 2.10 (t,
2 H, J ) 7.4 Hz), 1.48-1.40 (m, 4 H), 1.32-1.23 (m, 2 H), 0.98 (d,
3 H, J ) 6.3); 13C NMR (CD3OD) δ 177.6, 172.2, 170.0, 140.7, 140.4,
140.3, 140.2, 139.7, 139.6, 138.8, 132.5, 132.4, 131.2, 131.1, 130.0,
129.6, 129.5, 129.4, 129.4, 129.3, 129.3, 129.1, 129.1, 129.0, 128.8,
128.7, 128.6, 128.5, 128.5, 128.4, 128.3, 103.7, 101.3, 97.2, 81.8, 80.2,
78.7, 77.2, 76.6, 76.3, 76.2, 75.7, 75.4, 75.3, 74.3, 74.0, 73.5, 73.1,
70.5, 69.4, 67.1, 60.3, 34.9, 30.6, 26.8, 25.9, 17.2; MALDI-TOF MS
m/z calcd for C81H89NO19 1379, found 1402 (M + Na)+; LR-FAB MS
m/z calcd for C81H89NO19 (M) 1379.6, found 1423.3 (M + 2Na - H)+.
Typical Procedure for the N-Acetylation Reaction. A solution of
compound 26b (30 mg, 0.022 mmol) in 1-butanol-ethylenediamine
9
3598 J. AM. CHEM. SOC. VOL. 124, NO. 14, 2002