6316
Y.J. Lee et al. / Tetrahedron 65 (2009) 6310–6319
for 48 h, reaction mixture was concentrated in vacuo. The residure
was purified by column chromatography on Iatrobeads (CHCl3/
MeOH, 9:1, v/v) to afford compound 20 (65 mg, 0.052 mmol, 86%, a/
brine (2 mL), dried over MgSO4, and concentrated in vacuo. The
residue was purified by column chromatography on Iatrobeads
(CHCl3/MeOH, 9:1, v/v) to afford compound 22 (39 mg, 0.027 mmol,
87%).
b
¼2:1).
Rf¼0.38 (CHCl3/MeOH, 9:1, v/v); 1H NMR (400 MHz, CDCl3/
Rf¼0.48 (CHCl3/MeOH, 9:1, v/v); 1H NMR (400 MHz, CDCl3/
CD3OD, 3:1, v/v)
d
1.62 (s, 3H, Ac), 1.63 (s, 6H, 2ꢂAc), 1.64 (s, 3H, Ac),
CD3OD, 9:1, v/v) d 1.68 (s, 3H, Ac), 1.82 (s, 3H, Ac), 1.83 (s, 3H, Ac),
1.71 (s, 3H, Ac), 1.72 (s, 3H, Ac), 1.73 (s, 3H, Ac), 1.74 (s, 3H, Ac), 1.75
(s, 3H, Ac),1.79 (s, 3H, Ac),1.83 (s, 3H, Ac),1.85 (s, 3H, Ac),1.86 (s, 3H,
Ac),1.91 (s, 3H, Ac), 3.28–3.36 (m, 2H), 3.38–3.55 (m, 7H), 3.65–3.87
(m, 5H), 4.03–4.14 (m, 5H), 4.22–4.27 (m, 2H), 4.38–4.41 (m, 1H),
4.71 (t, J¼10.0 Hz, 1H), 4.78–4.85 (m, 2H), 4.90–4.95 (m, 1H), 4.98 (t,
J¼10.1 Hz, 1H), 5.35 (d, J¼8.7 Hz, 0.33H, H-1GlcNb), 5.75 (d, J¼3.7 Hz,
1.84 (s, 3H, Ac),1.91 (s, 3H, Ac),1.93 (s, 9H, 3ꢂAc),1.95 (s, 6H, 2ꢂAc),
2.00 (s, 3H, Ac), 2.07 (s, 6H, 2ꢂAc), 3.46–3.52 (m, 2H), 3.57–3.71 (m,
5H), 3.76–3.81 (m, 2H), 3.87–4.11 (m, 5H), 4.14–4.24 (m, 4H), 4.28–
4.31 (m, 1H), 4.32 (d, J¼8.2 Hz, 1H), 4.38 (d, J¼8.2 Hz, 1H), 4.47 (d,
J¼8.7 Hz, 1H), 4.90–5.02 (m, 7H), 5.03–5.08 (m, 1H), 5.11 (t,
J¼10.1 Hz, 1H), 5.51 (dd, J¼6.0, 3.2, H-1GlcNa), 7.23–7.32 (m, 10H, Ar–
0.67H, H-1GlcNa); 20
a
:
13C NMR (100 MHz, CDCl3/CD3OD, 3:1, v/v)
H); 13C NMR (100 MHz, CDCl3/CD3OD, 9:1, v/v)
d 20.3(4), 20.4, 20.47,
d
19.8, 19.9, 20.0 (2), 20.02 (2), 20.1 (2), 20.12 (2), 20.2 (2), 22.0 (2),
20.5, 20.6 (2), 22.2, 22.5 (3), 51.6 (d, JC–P¼8.6 Hz, C-2), 53.9, 53.94,
54.4, 61.7, 62.5, 62.6, 68.2, 69.6, 69.96, 70.0, 70.2, 70.3, 71.5, 72.3,
72.5, 72.6, 72.7, 72.9, 75.3, 75.6, 75.7, 96.0 (d, JC–P¼6.7 Hz, C-1),
100.8, 100.9, 100.91, 128.0, 128.7, 128.73, 128.9, 134.9 (d, JC–
50.4, 54.0, 54.1, 54.5, 61.4, 61.7, 62.3, 62.4, 68.0, 70.2, 70.6, 71.1, 71.9,
72.15, 72.2, 72.5, 72.6, 75.36, 75.4, 75.5, 90.0, 100.2, 100.3, 100.5,
169.3, 169.6, 170.2, 170.37, 170.4, 170.6, 170.7, 171.0 (2), 171.2 (2),
171.5, 171.54, 171.6; 20
b
:
13C NMR (100 MHz, CDCl3/CD3OD, 3:1, v/v)
¼7.6 Hz), 135.0 (d, JC–P¼6.7 Hz), 169.7, 170.5, 170.7, 170.8 (2), 170.9,
P
d
91.7, 100.0, 100.2, 100.3. MALDI-TOF MS: [MþNa]þ calcd for
171.2, 171.3 (2), 171.4, 171.44 (3); 31P NMR (162 MHz, CDCl3/CD3OD,
C52H74N4NaO31, 1273.4, found 1273.3; HRMS ESI-TOF: [MþNa]þ
9:1, v/v)
d
ꢀ2.1. MALDI-TOF MS: [MþNa]þ calcd for
calcd for C52H74N4NaO31, 1273.4235, found 1273.4227.
C64H85N4NaO33P, 1491.5, found 1491.4; HRMS ESI-TOF: [MþNa]þ
calcd for C64H85N4NaO33P, 1491.4731, found 1491.4699.
4.15. 2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-
glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-acetyl-2-deoxy-
-glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-acetyl-2-
deoxy- -glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-
acetyl-2-deoxy- -glucopyranose (21)
b-D-
4.17. 2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-
glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-acetyl-2-deoxy-
-glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-acetyl-2-
deoxy- -glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-
acetyl-2-deoxy- -glucopyranosyl phosphate (23)
b-D-
b-D
b-D
b-D
D
b-D
a-D
The mixture of acetylated tetra-N-acetylchitotetraose 20
(216 mg, 0.173 mmol) and powdered, activated 4 Å molecular
sieves (300 mg) in MeOH (3 mL) and CH2Cl2 (3 mL) was stirred at
room temperature for 5 h. The reaction mixture was neutralized
with Dowex CCR-3 (Hþ mode) resin, filtered through CeliteÒ, and
concentrated. The residue was purified by column chromatography
on Iatrobeads (CHCl3/MeOH, 9:1, v/v) to afford compound 21
(128 mg, 0.106 mmol, 61%).
The mixture of dibenzyl phosphate 22 (51 mg, 0.035 mmol) and
Pd/C (10%, 50 mg) in MeOH (2 mL) and CH2Cl2 (1 mL) under H2
(1 atm) atmosphere was stirred at room temperature for 3 h. The
reaction mixture was filtered through CeliteÒ and washed with
MeOH. Et3N (1 mL) was added followed by concentration in vacuo
to give the mono (triethylammonium) salt of phosphate 23 (47 mg,
0.034 mmol, 97%).
Rf¼0.25 (CHCl3/MeOH, 9:1, v/v); 1H NMR (400 MHz, CDCl3/
Rf¼0.18 (CHCl3/MeOH/H2O, 70:30:5, v/v/v); 1H NMR (400 MHz,
CD3OD, 3:1, v/v)
d
1.82 (s, 3H, Ac), 1.83 (s, 3H, Ac), 1.84 (s, 3H, Ac),
CDCl3/CD3OD, 1:1, v/v)
d
1.25 (t, J¼7.3 Hz, 9H, N(CH2CH3)3), 1.87 (s,
1.89 (s, 6H, 2ꢂAc), 1.90 (s, 3H, Ac), 1.92 (s, 6H, 2ꢂAc), 1.93 (s, 3H,
Ac), 1.99 (s, 3H, Ac), 2.04 (s, 3H, Ac), 2.05 (s, 3H, Ac), 2.06 (s, 3H, Ac),
3.53–3.62 (m, 5H), 3.73 (t, J¼9.6 Hz, 1H), 3.78–3.87 (m, 2H), 3.90–
3.94 (m, 1H), 3.98–4.13 (m, 5H), 4.17–4.31 (m, 7H), 4.42 (d, J¼8.7 Hz,
1H), 4.88–4.94 (m, 3H), 5.01 (d, J¼3.7 Hz, H-1GlcNa), 5.07 (t,
J¼10.1 Hz, 1H), 5.30 (t, J¼10.1 Hz, 1H); 13C NMR (100 MHz, CDCl3/
3H, Ac), 1.88 (s, 3H, Ac), 1.89 (s, 3H, Ac), 1.91 (s, 3H, Ac), 1.96 (s, 3H,
Ac), 1.97 (s, 3H, Ac), 1.98 (s, 6H, 2ꢂAc), 2.02 (s, 3H, Ac), 2.05 (s, 3H,
Ac), 2.09 (s, 3H, Ac), 2.11 (s, 3H, Ac), 2.12 (s, 3H, Ac), 3.01 (q, J¼7.3 Hz,
6H, N(CH2CH3)3), 3.59–3.76 (m, 8H), 3.81 (t, J¼10.1 Hz, 1H), 3.98–
4.16 (m, 6H), 4.33–4.42 (m, 3H), 4.49–4.52 (m, 1H), 4.57 (d,
J¼8.7 Hz, 1H), 4.65 (d, J¼8.2 Hz, 1H), 4.73 (d, J¼8.7 Hz, 1H), 4.95 (t,
J¼9.9 Hz, 1H), 5.12–5.19 (m, 2H), 5.21 (t, J¼10.6 Hz, 1H), 5.30 (t,
J¼10.1 Hz, 1H), 5.40 (dd, J¼6.8, 3.7 Hz, H-1GlcNa); 13C NMR
CD3OD, 3:1, v/v) d 20.1, 20.2 (3), 20.23 (2), 20.3, 20.4 (2), 20.43, 22.2,
22.3, 22.31, 51.9, 53.9 (2), 54.4, 61.5, 62.4, 62.5 (2), 67.7, 68.1, 71.2,
71.3, 72.0, 72.2, 72.3 (2), 72.5, 75.5, 75.8, 76.0, 90.9, 100.5, 100.6,
100.9, 169.6, 170.4, 170.5, 170.6, 170.7, 170.9, 171.1 (2), 171.3, 171.36,
171.4, 171.5, 172.0. MALDI-TOF MS: [MþNa]þ calcd for
C50H72N4NaO30, 1231.4, found 1231.4; HRMS ESI-TOF: [MþNa]þ
calcd for C50H72N4NaO30, 1231.4129, found 1231.4084.
(100 MHz, CDCl3/CD3OD,1:1, v/v) d 8.91, 20.6, 20.7, 20.8, 20.88, 20.9,
21.0 (4), 22.6, 22.8, 22.85, 22.9, 46.8, 52.7 (d, JC–P¼7.6 Hz, C-2), 55.3,
55.5, 55.6, 59.2, 62.4, 62.6, 63.3, 63.5, 69.1, 69.8, 72.1, 72.5, 72.8,
73.05, 73.1, 73.5, 76.3, 76.5, 76.7, 94.4 (d, JC–P¼5.7 Hz, C-1), 101.0,
101.1 (2), 170.7, 171.2, 171.3 (2), 171.5, 171.7, 171.9, 172.0, 172.1, 172.6,
172.7, 172.73, 127.8; 31P NMR (162 MHz, CDCl3/CD3OD, 1:1, v/v)
4.16. Dibenzyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-
glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-acetyl-2-deoxy-
-glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-acetyl-2-
deoxy- -glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-
acetyl-2-deoxy- -glucopyranosyl phosphate (22)
b-
D-
d
ꢀ0.9. MALDI-TOF MS: [MꢀH]ꢀ calcd for C50H72N4O33P, 1287.4,
found 1287.4; HRMS ESI-TOF: [MꢀH]ꢀ calcd for C50H72N4O33P,
b-D
1287.3816, found 1287.3858.
b-D
a-
D
4.18. Undecaprenyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-
b
-
D
-glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-acetyl-2-
deoxy- -glucopyranosyl-(1/4)-2-acetamido-3,6-di-O-
acetyl-2-deoxy- -glucopyranosyl-(1/4)-2-acetamido-3,6-
di-O-acetyl-2-deoxy- -glucopyranosyl diphosphate (24)
To a solution of hemiacetal 21 (38 mg, 0.031 mmol) in DMF
(1 mL) was added dropwise lithium hexamethyldisilazide (1 M
b-D
b-D
solution in THF, 40
m
L, 0.040 mmol) at ꢀ50 ꢁC. The mixture was
a-D
stirred for 10 min and then a solution of tetrabenzyl pyrophosphate
(25 mg, 0.046 mmol) in THF (0.3 mL) was added. After stirring at
ꢀ50 ꢁC for further 1 h, the reaction mixture was allowed to warm
up to 0 ꢁC over 30 min. The mixture was diluted with CHCl3
(10 mL), washed with saturated aqueous NaHCO3 (2ꢂ2 mL) and
To a solution of phosphate 23 (12 mg, 8.6
was added carbonyl diimidazole (CDI) (4 mg, 24.7
stirring at room temperature for 4 h, the excess CDI was quenched
with MeOH (3 L) and stirred for 1 h. The reaction mixture was
mmol) in DMF (1 mL)
mmol). After
m