11072 J. Am. Chem. Soc., Vol. 122, No. 45, 2000
Ruda et al.
Hz), 1.69 (s, 3H), 1.99 (s, 6H), 2.50-2.70 (m, 2H), 3.45-4.97 (m,
56H), 5.00 (s, 1H), 5.14 (s, 1H), 5.23 (s, 1H), 5.30-5.40 (m, 1H),
5.40 (s, 1H), 7.00-7.40 (m, 60H); HRMS Calcd for C124H135O29ClS:
[M + Na]+ 2177.8396. Found: [M + Na]+ 2177.8315.
74.5, 74.6, 74.7, 74.8, 75.0, 75.2, 75.9, 76.1, 76.1, 76.7, 77.8, 77.9,
78.0, 78.1, 78.9, 79.4, 80.1, 80.4, 82.0, 84.2, 97.2, 99.1, 99.2, 100.0,
100.2, 102.1, 112.4, 119.0, 126.9-128.6, 135.5, 135.7, 137.8-138.9,
1
169.5, 170.1, 170.2; H, δ 0.84 (s, 3H), 0.89 (s, 3H), 0.98 (s, 3H),
2,3,4,6-Tetra-O-benzyl-r-D-galactopyranosyl-(1f6)-2,3,4-tri-O-
benzyl-r-D-galactopyranosyl-(1f3)-2,5,6-tri-O-acetyl-â-D-galacto-
furanosyl-(1f3)-2,4-di-O-benzyl-6-O-chloroacetyl-r-D-mannopyr-
anosyl-(1f3)-2,4,6-tri-O-benzyl-r-D-mannopyranosyl-(1f4)-2-azido-
3,6-di-O-benzyl-2-deoxy-r-D-glucopyranosyl-(1f6)-1-O-
dibenzylphosphoryl-4,5-O-isopropylidene-2,3-O-(D-1,7,7-
trimethyl[2.2.1]bicyclohept-6-ylidene)-D-myo-inositol (12a). To a
stirred mixture of 10 (167 mg, 0.077 mmol), 113 (83 mg, 0.085 mmol),
and 4 Å molecular sieves in diethyl ether (30 mL) under argon was
added DMTST (60 mg, 0.233 mmol). After 18 h Et3N (0.2 mL) was
added, and the mixture was filtered through Celite. Evaporation of the
solvent and FC (toluene/EtOAc 9:1, 0.25% Et3N) of the residue yielded
12a (179 mg, 0.058 mmol, 75%). Rf 0.44 (toluene/EtOAc 4:1); [R]D
+44 (c 1.0, CHCl3); NMR (CDCl3): 13C, δ 10.0, 20.3, 20.5, 20.6, 20.7,
26.9, 27.1, 29.7, 40.7, 43.9, 45.2, 48.0, 51.5, 62.5, 62.8 64.8, 66.0,
68.7, 68.9, 69.5, 69.6, 69.7, 69.8, 70.4, 70.5, 71.6, 72.2, 72.4, 72.6,
72.9, 73.0, 73.2, 73.4, 73.5, 73.7, 73.8, 73.9, 74.2, 74.3, 74.4, 74.6,
74.7, 75.2, 75.4, 76.0, 76.3, 76.8, 77.4, 77.6, 78.7, 78.9, 79.2, 79.5,
80.3, 82.6, 83.9, 97.3 (JC,H ) 175 Hz), 98.3 (JC,H ) 172 Hz), 99.0
(JC,H ) 169 Hz), 100.3 (JC,H ) 174 Hz), 100.4 (JC,H ) 175 Hz), 101.9
(JC,H ) 173 Hz), 112.5, 119.1, 127.0-130.0, 135.6, 137.7-138.9, 166.9,
169.5, 170.1, 170.2;1H, δ 0.83 (s, 3H), 0.89 (s, 3H), 0.98 (s, 3H), 1.10-
1.41 (m, 9H), 1.60-2.10 (m, 4H), 1.80 (s, 3H), 1.93 (s, 3H), 1.96 (s,
3H), 3.22 (dd, 1H, J ) 9.8, 3.4 Hz), 3.40-5.17 (m, 79H), 5.27-5.35
(m, 1H), 5.46 (s, 1H), 7.02-7.38 (m, 80H); HRMS Calcd for
1.10-1.45 (m, 9H), 1.60-2.10 (m, 4H), 1.83 (s, 3H), 1.91 (s, 3H),
1.98 (s, 3H), 3.22 (dd, 1H, J ) 9.8, 3.6 Hz), 3.37 (dd, 1H, J ) 8.8, 5.8
Hz), 3.43-5.17 (m, 76H), 5.27-5.35 (m, 1H), 5.36 (s, 1H), 7.02-
7.38 (m, 80H); 31P (decoupled),
δ
-0.97; HRMS Calcd for
173H192O41N3P: [M + 1 + Na]+ 3022.2701. Found: [M + 1 + Na]+
3022.2733.
C
2,3,4,6-Tetra-O-benzyl-r-D-galactopyranosyl-(1f6)-2,3,4-tri-O-
benzyl-r-D-galactopyranosyl-(1f3)-2,5,6-tri-O-acetyl-â-D-galacto-
furanosyl-(1f3)-[2,3,4,6-tetra-O-benzyl-r-D-glucopyranosyl-(1-PO4-
6)-2,4-di-O-benzyl-r-D-mannopyranosyl]-(1f3)-2,4,6-tri-O-benzyl-
r-D-mannopyranosyl-(1f4)-2-azido-3,6-di-O-benzyl-2-deoxy-r-D-
glucopyranosyl-(1f6)-1-O-dibenzylphosphoryl-4,5-O-isopropylidene-
2,3-O-(D-1,7,7-trimethyl[2.2.1]bicyclohept-6-ylidene)-D-myo-
inositol triethylammonium salt (14a). 12b (48 mg, 0.016 mmol) and
134,7 (24 mg, 0.034 mmol) were dissolved in CH3CN/pyridine (3:1, 4
mL), and pivaloyl chloride (20 µL, 0.16 mmol) was added. After 25
min I2 (2% in pyridine/water 49:1, 1.0 mL, 0.079 mmol) was added.
After an additional 30 min, the reaction mixture was diluted with CHCl3
and washed with 10% Na2S2O3 (aq). Normal workup and FC (CHCl3/
MeOH 100:1f25:1, 0.25% Et3N) gave 14a (56 mg, 0.015 mmol, 94%).
Rf 0.62 (CHCl3/MeOH 9:1); [R]D +37 (c 1.1, CHCl3); NMR (CDCl3):
13C, δ 8.7, 10.0, 20.2, 20.5 (2C), 20.6, 20.7, 27.0 (2C), 27.3, 29.9,
43.7, 45.2, 48.0, 51.5, 62.8, 63.4, 64.3, 66.0, 68.4, 68.8, 69.0, 69.5,
69.6, 69.7, 70.5, 71.2, 71.7, 71.9, 72.5, 72.5, 72.6, 72.8, 73.1, 73.4,
73.5, 73.8, 73.9, 74.2, 74.5, 74.6, 74.7, 74.9, 75.3, 75.5, 75.7, 76.0,
76.2, 77.3, 77.8, 77.9, 78.0, 78.2, 78.8, 79.0, 79.5, 79.8, 79.9, 80.2,
81.6, 82.9, 83.6, 84.1, 93.1 (d, J ) 7.4 Hz), 97.4, 98.5, 99.1, 100.4,
100.5, 102.4, 112.4, 119.0, 127.0-128.6, 135.5, 135.7, 137.9-139.0,
169.6, 170.0, 170.3; 1H, δ 0.83 (s, 3H), 0.90 (s, 3H), 0.97 (s, 3H), 1.00
(t, 9H, J ) 7.0 Hz), 1.10-1.45 (m, 9H), 1.60-2.10 (m, 13H), 2.61
(m, 6H), 3.22 (dd, 1H, J ) 9.8, 3.0 Hz), 3.33-5.25 (m, 92H), 5.46 (s,
1H), 5.94 (dd, 1H, J ) 8.2, 3.1 Hz), 7.02-7.38 (m, 100H); 31P
(decoupled), δ -1.06, -0.66; HRMS Calcd for C207H227O49N3P2: [M
+ 2 + Na]+ 3625.4804. Found: [M + 2 + Na]+ 3625.4752.
r-D-Galactopyranosyl-(1f6)-r-D-galactopyranosyl-(1f3)-â-D-ga-
lactofuranosyl-(1f3)-[r-D-glucopyranosyl-(1-PO4-6)-r-D-mannopy-
ranosyl]-(1f3)-r-D-mannopyranosyl-(1f4)-2-amino-2-deoxy-r-D-
glucopyranosyl-(1f6)-D-myo-inositol 1-hydrogenphosphate, Ammon-
ium Salt (1b). To a stirred solution of 14a (54 mg, 0.015 mmol) in
CH2Cl2/MeOH 2:1 (6 mL) was added NaOMe (25 mg). After 2 h the
mixture was diluted with CH2Cl2 (50 mL) and washed with 1 M NH4-
Cl. Normal workup and FC (CHCl3/MeOH 25:1 with 0.25% Et3N) gave
14b (42 mg, 0.012 mmol, 80%). Rf 0.55 (CHCl3/MeOH 9:1). Depro-
tection of this derivative by the same procedure as for 12a gave 1b
(16 mg, 0.011 mmol, 90%); [R]D +88 (c 0.75, H2O); NMR (D2O) pH*
) 5.6: 13C, δ 54.7, 60.8, 61.1, 61.5, 62.0 (2C), 63.6, 66.0 (d, J ) 7.4
Hz), 66.3, 67.6, 67.8, 69.0 (2C), 69.9 (2C), 70.0, 70.2 (2C), 70.3, 70.6,
71.3, 71.4, 71.8 (2C), 72.0, 72.1, 72.5, 73.0, 73.1, 73.3, 73.3, 73.6,
74.6, 75.9, 76.0, 76.5 (d, J ) 5.5 Hz), 77.4 (d, J ) 3.7 Hz), 80.0, 80.1,
82.8, 85.6, 95.7, 96.1 (d, J ) 5.5 Hz), 99.3, 100.4, 102.3, 103.4, 105.6;
1H, δ 3.33 (dd, 1H, J ) 10.6, 4.0 Hz), 3.44 (t, 1H, J ) 9.3 Hz), 3.49-
4.38 (m, 45H), 4.44 (br, 1H), 4.98 (d, 1H, J ) 3.3 Hz), 5.06 (d, 1H, J
) 3.6 Hz), 5.12 (s, 1H), 5.20 (s, 1H), 5.30 (d, 1H, J ) 1.1 Hz), 5.53
(dd, 1H, J ) 7.3, 3.3 Hz), 5.69 (d, 1H, J ) 3.7 Hz); 31P (decoupled),
δ -1.15, 1.76; HRMS Calcd for C48H85NO46P2: [M - H + 2Na]+
1518.3536. Found: [M - H + 2Na]+ 1518.3538.
C
175H193O42N3ClP: [M + Cs]+ 3207.1539. Found: [M + Cs]+
3207.1584.
(r-D-Galactopyranosyl)-(1f6)-(r-D-galactopyranosyl)-(1f3)-
(â-D-galactofuranosyl)-(1f3)-(r-D-mannopyranosyl)-(1f3)-(r-
D-mannopyranosyl)-(1f4)-(2-amino-2-deoxy-r-D-glucopyranosyl)-
(1f6)-D-myo-inositol 1-phosphate, ammonium salt (1a). To a stirred
solution of 12a (38 mg, 0.0098 mmol) in CH2Cl2/MeOH 2:1 (6 mL)
was added NaOMe (11 mg). After 5 h the mixture was diluted with
CH2Cl2 (25 mL), washed with water, dried, filtered, and concentrated.
The residue was dissolved in THF (2 mL) and added to NH3 (l) ( ∼20
mL) at -33 °C. To the stirred mixture was added a minimum amount
of sodium for the mixture to turn deep blue. After 1 min NH4Cl was
added until the color disappeared. The mixture was concentrated, and
the residue was dissolved in 0.1 M HCl (10 mL). After 6 h the mixture
was neutralized with NH3 (25%, 0.1 mL), washed with ether (10 mL),
and concentrated. Gel filtration of the residue on a Pharmacia Sephadex
G-15 column eluted with H2O containing 1% n-butanol afforded 1a
(9.4 mg, 0.0076 mmol, 78%). [R]D +86 (c 1.0, H2O); NMR (D2O)
pH* ) 5.8: 13C, δ 54.6, 60.8, 61.5, 62.0 (2C), 63.6, 66.1, 66.7, 67.7,
67.8, 68.9, 69.0, 69.9, 70.0, 70.2 (2C), 70.3, 70.6, 71.3, 71.4 (2C),
71.8 (2C), 72.6, 73.1, 73.6, 74.2, 74.6, 76.1 (2C), 76.3, 78.4 (d, J ) 6
Hz), 78.6, 80.2, 82.8, 85.4, 96.0, 99.3, 100.3, 102.1, 102.8, 105.6; 1H,
δ 3.33 (dd, 1H, J ) 10.6, 4.0 Hz), 3.40 (t, 1H, J ) 9.3 Hz), 3.56 (dd,
1H, J ) 10.1, 2.7 Hz), 3.62-4.35 (m, 38H), 4.43 (br, 1H) 4.96 (d, 1H,
J ) 3.3 Hz), 5.06 (d, 1H, J ) 3.3 Hz), 5.16 (d, 1H, J ) 1.5 Hz), 5.19
(s, 1H), 5.27 (d, 1H, J ) 1.8 Hz), 5.67 (d, 1H, J ) 4.0 Hz); 31P
(decoupled), δ 1.99; HRMS Calcd for C42H74O38NP: [M - 1]-
1230.3548. Found: [M - 1]- 1230.3579.
2,3,4,6-Tetra-O-benzyl-r-D-galactopyranosyl-(1f6)-2,3,4-tri-O-
benzyl-r-D-galactopyranosyl-(1f3)-2,5,6-tri-O-acetyl-â-D-galacto-
furanosyl-(1f3)-2,4-di-O-benzyl-r-D-mannopyranosyl-(1f3)-2,4,6-
tri-O-benzyl-r-D-mannopyranosyl-(1f4)-2-azido-3,6-di-O-benzyl-
2-deoxy-r-D-glucopyranosyl-(1f6)-1-O-dibenzylphosphoryl-4,5-O-
isopropylidene-2,3-O-(D-1,7,7-trimethyl[2.2.1]bicyclohept-6-ylidene)-
D-myo-inositol (12b). To 12a (57 mg, 0.019 mmol) in CH2Cl2 (3 mL)
was added MeOH saturated with NH3 (6 mL). After 50 min the mixture
was concentrated. FC (toluene/EtOAc 6:1, 0.25% Et3N) of the residue
gave 12b (49 mg, 0.016 mmol, 88%). Rf 0.34 (toluene/EtOAc 4:1);
[R]D +41 (c 0.84, CHCl3); NMR (CDCl3):13C, δ 10.0, 20.3, 20.5, 20.5,
20.7 (2C), 26.9 (2C), 27.1, 29.8, 43.9, 45.1, 48.0, 51.5, 62.1, 62.6,
63.1, 66.1, 68.5, 68.8, 69.0, 69.3, 69.5, 69.6, 69.7, 69.8, 70.5, 71.7,
72.3, 72.5, 73.0, 73.1, 73.2, 73.3, 73.4, 73.5, 73.7, 73.8, 74.0, 74.1,
Acknowledgment. This work, which was part of a col-
laboration with Dr. Salvatore Turco Department of Biochem-
istry, University of Kentucky, Lexington, USA and Dr. Hannah
Akuffo, Microbiology and Tumor Biology Centre, Karolinska
Institute, Stockholm, Sweden, was supported by the Swedish
Natural Science Research Council.
JA001515T