S. Sato et al. / Carbohydrate Research 339 (2004) 2425–2432
2431
CHCl3–MeOH) to give 1-a (93mg, 90%) as a colorless
solid.
The preparation of the b-anomer was carried out in
the same manner as that used for the a-anomer, using
5% Pd–C.
NH–), 6.5 (0.5H, br s, enol OH); FABMS (NBA, m/z):
749 (M+H)+; IR (KBr) m: 3340, 2931, 2869, 1774,
1668, 1514, 1363, 1203, and 1072cmꢀ1. Anal. Calcd
for C45H52N2O8: C, 72.17; H, 7.00; N, 3.74. Found: C,
72.19; H, 7.01; N, 3.66.
1.6.1. Data for the a-anomer. Colorless amorphous
1.7.2. Data for the b-anomer. Colorless prisms; Rf 0.34
19
D
powder; Rf 0.56 (5:1 CHCl3–MeOH); ½a þ 98:8 (c
(2:1 hexane–EtOAc); mp 122–125ꢁC (from diethyl
24
1
1.00, MeOH); H NMR (DMSO-d6) d: 0.97 and 0.98
ether–diisopropyl ether); ½a ꢀ 27:9 (c 1.005, CHCl3);
D
(each 3H, s, CH3 · 2), 1.13 (6H, s, CH3 · 2), 1.40 and
1.50 (each 1H, t, J 12.2Hz, CH2), 1.95 (2H, m, CH2),
2.04 (3H, s, OAc), 3.03 (1H, dt, J 5.6 and 9.0Hz, H-
40), 3.16 (1H, m, J 3.9, 6.8, and 9.0Hz, H-20), 3.37
(1H, dt, J 4.6 and 9.0Hz, H-30), 3.43 (2H, m, H-50 and
60a), 3.62 (1H, dd, J 5.6 and 9.5Hz, H-6b0), 3.91 (1H,
m, H-4), 4.51 (1H, t, J 5.6Hz, 60-OH),. 4.55 (1H, t, J
6.8Hz, 20-OH), 4.75 (1H, t, J 4.6Hz, 30-OH), 4.79
(1H, t, J 3.9Hz, H-10), 4.87 (1H, t, J 5.6Hz, 40-OH);
FABMS (glycerol, m/z): 578 (M+H)+; IR (KBr) m:
3400, 2975, 2931, 1766, 1367, 1251, 1203, 1188, 1049,
and 1028cmꢀ1. Anal. Calcd for C17H31NO8Æ0.25H2O:
C, 53.46; H, 8.31; N, 3.67. Found: C, 53.56; H, 8.44;
N, 3.63.
1H NMR (CDCl3, at 40ꢁC) d: 1.28 (6H, s, CH3 · 2),
1.39 (6H, s, CH3 · 2), 2.13 (3H, s, OAc), 3.39 (1H, t, J
9.0Hz, H-20), 3.52 (1H, d, J 9.0Hz), 3.69–3.79 (4H,
m), 4.46 and 4.59 (each 1H, d, J 11Hz, benzylic CH2),
4.69 and 4.78 (each 1H, d, J 11Hz, benzylic CH2),
4.54 and 4.81 (each 1H, d,
J 11Hz, benzylic
CH2), 4.89 (2H, s, benzylic CH2), 5.15 (1H, t, J 9.0Hz,
H-10), 5.66 and 5.76 (total 1H, br s, olefinic H), 5.88
(1H, br s, NH), 7.25–7.33 (20H, m, CH2Ph · 4);
FABMS (NBA, m/z): 749 (M+H)+; IR (KBr) m: 3346,
2931, 2868, 1774, 1676, 1531, 1363, 1205, and
1070cmꢀ1. Anal. Calcd for C45H52N2O8: C, 72.17; H,
7.00; N, 3.74. Found: C, 72.00; H, 6.84; N, 3.76.
1.8. N1-Acetoxy-2,2,5,5-tetramethylpyrrolin-3-oyl a-D-
glucopyranosylamine (2-a)
1.6.2. Data for the b-anomer. Colorless viscous oil; Rf
19
0.56 (5:1 CHCl3–MeOH); ½a ꢀ 33:0 (c 0.995, MeOH);
D
1H NMR (DMSO-d6) d: 0.97 and 0.98 (each 3H, s,
CH3 · 2), 1.13 (6H, s, CH3 · 2), 1.40 and 1.47 (each
1H, t, J 12.2Hz, CH2), 1.96 (2H, m, CH2), 2.04 (3H, s,
OAc), 2.90 (1H, br t, J 7.5 and 9.0Hz, H-20), 3.02 (1H,
t, J 9.0Hz, H-40), 3.11 (1H, t, J 9.0Hz, H-30), 3.13 (1H,
m, H-50), 3.45 (1H, dd, J 6.5 and 11.5Hz, H-6a0), 3.65
(1H, d, J 11.5Hz, H-6b0), 3.97 (1H, m, H-4), 4.23 (1H,
d, J 7.5Hz, H-10), 4.47 (1H, t, J 5.7Hz, 60-OH),. 4.90
(1H, t, J 4.6Hz, 40-OH), 4.92 (1H, d, J 4.8Hz, 30-OH),
4.94 (1H, t, J 4.4Hz, 20-OH); FABMS (glycerol, m/z):
578 (M+H)+; IR (KBr) m: 3400, 2974, 2931, 1766, 1367,
1253, 1205, 1188, 1076, 1049, and 1028cmꢀ1. Anal. Cal-
cd for C17H31NO8Æ0.25H2O: C, 53.46; H, 8.31; N, 3.67.
Found: C, 53.68; H, 8.62; N, 3.63.
The preparation was carried out in the same manner as
that used for 1, using 10% Pd–C. Colorless amorphous
23
powder; Rf 0.42 (3:1 CHCl3–MeOH); ½a þ 68:1 (c
D
1.045, MeOH); 1H NMR (DMSO-d6) d: (pyrroline moi-
ety) 1.16 (6H, s, CH3 · 2), 1.21 and 1.25 (each 3H, s,
CH3 · 2), 2.09 (3H, s, OAc), 6.53 (1H, s, olefinic H),
7.80 (1H, br d, J 8.5Hz, NH); (sugar moiety) 3.1, 3.2,
3.4, 3.5, 3.6, and 3.7 (each 1H, m), 4.5 (1H, br m, 60-
OH), 4.81 (1H, d, J 4.2Hz, OH), 4.92 (1H, d, J 4.6Hz,
OH), 5.06 (1H, d, J 4.6Hz, OH), 5.47 (1H, m, H-10);
FABMS (NBA, m/z): 389 (M+H)+; IR (KBr) m: 3417,
2983, 2933, 1770, 1754, 1668, 1621, 1519, 1211, and
1061cmꢀ1. Anal. Calcd for C17H28N2O8Æ1.25H2O : C,
49.69; H, 7.48; N, 6.82. Found: C, 49.89; H, 7.12; N, 6.48.
1.7. N1-Acetoxy-2,2,5,5-tetramethylpyrrolin-3-oyl
1.9. Enzyme-catalyzed hydrolysis of 1-a, 1-b, and 2-a
2,3,4,6-tetra-O-benzyl-a- and -b-D-glucopyranosylamine (9)
Porcine liver esterase (40lL) was added to a PBS buffer
solution (pH8.0, 1mL) of 1 (2mg), and the mixture was
shaken (120rpm) at 25ꢁC for 1day. The progress of the
reaction was monitored by silica-gel TLC (0.25mm sil-
ica-gel F254 plates (E. Merck), solvent system: 5:1
CHCl3–MeOH), which was observed by the emission
using UV 254nm light-irradiation and the coloring
using a 5% ethanolic solution of phosphomolybdic acid
with heat. The enzymes used included the following:
Porcine liver esterase (Sigma Chemical Co.); CAL (No-
vozyme 435 and 525Lꢂ, Novo Nordisk Bio Industry
Co., and Chirazyme L-2ꢂ, Boehringer Mannheim
Co.); Lipase AH, PS, AY, and AK (Amano Pharmaceu-
1.7.1. Data for the a-anomer. Colorless amorphous
26
D
powder; Rf 0.47 (2:1 hexane–EtOAc); ½a þ 50:9 (c
1
1.08, CHCl3); H NMR (CDCl3, at 40ꢁC) d: 1.28 (6H,
s, CH3 · 2), 1.39 (6H, br s, CH3 · 2), 2.13 (3H, s,
OAc), 3.63–3.69 (3H, m, H-30, 40, and 50), 3.73 (1H, br
d, J 9.5Hz, H-60a), 3.77 (1H, dd, J 2.7 and 9.5Hz, H-
60b), 3.85 (1H, dd, J 5.2 and 9.1Hz, H-20), 4.58 (2H, s,
benzylic CH2), 4.49 and 4.61 (each 1H, d, J 11Hz, benz-
ylic CH2), 4.51 and 4.77 (each 1H, d, J 11Hz, benzylic
CH2), 4.80 and 4.92 (each 1H, d, J 11Hz, benzylic
CH2), 5.80 (1H, t, J 5.2Hz, H-10), 5.58 and 6.05 (each
0.5H, br s, olefinic H), 6.4 (0.5H, br d, J 5.2Hz, >