in THF was refluxed for 2 h. After the solution had cooled to
rt, freshly made amine (1.0 eq.) was added and reflux was
resumed for 22 h. The solution was cooled to rt and con-
centrated in vacuo. The resultant yellow oil was partitioned
between water and ethyl acetate; the organic phase was dried
(Na2SO4) and concentrated. Flash chromatography of the
residue with CH2Cl2–MeOH 30 : 1 afforded the product.
3H), 2.02 (s, 3H), 1.94 (s, 3H), 1.78 (m, 2H), 1.69–1.57 (m, 8H),
1.47 (m, 2H), 1.38 (m, 2H), 1.28 (m, 2H); δC(125 MHz; CDCl3)
176.5, 174.5, 173.3, 173.0, 171.5, 170.5 (m), 170.4 (m), 170.1,
170.0, 169.5, 168.9, 168.2, 161.1, 159.3, 159.0, 129.2, 119.7,
114.9, 101.3, 101.1, 93.7, 78.1, 75.9, 74.7, 73.1, 72.5, 71.2, 71.0,
69.8, 67.9, 67.4, 67.0, 66.7, 66.6, 64.9, 62.1, 61.4, 61.3, 39.5,
39.3, 36.7, 36.5, 29.4(3), 29.3(9), 27.3, 26.6, 26.4, 25.4, 24.8,
23.7, 21.1, 20.9(9) (m), 20.9(5) (m), 20.8(5) (m), 20.8, 20.7,
12.6; m/z (FAB MS) 1461.4 (Mϩ ϩ H. C66H89N6O31 requires
m/z, 1461.56); 1499.4 (Mϩ ϩ K. C66H88N6O31ؒK requires m/z,
1499.51).
Compound 13
From triethylene glycol monoazide (40 mg, 0.22 mmol), CDI
(40 mg, 0.24 mmol), THF (10 ml), and 9 (freshly prepared
from 106 mg, 0.1 mmol of 8) was obtained compound 13 (82
mg, 66%) as a white solid; δH(500 MHz; CDCl3) 6.17 (d, J = 9.0
Hz, 1H), 5.43 (d, J = 2.0 Hz, 1H), 5.32–5.21 (m, 4H), 5.19
(t, J = 9.2 Hz, 1H), 5.14 (dd, J = 10.8, 7.8 Hz, 1H), 5.07 (dd,
J = 10.8, 3.2 Hz, 1H), 4.84 (m, 1H), 4.80 (t, J = 9.5 Hz, 1H),
4.39–4.36 (m, 2H), 4.20–4.00 (m, 8H), 3.82–3.60 (m, 14H), 3.38
(t, J = 5.0 Hz, 2H), 3.13 (q, J = 6.5 Hz, 2H), 2.14 (s, 3H), 2.12 (s,
6H), 2.08 (s, 3H), 2.06 (s, 3H), 2.05 (s, 3H), 2.04 (s, 3H), 2.03 (s,
3H), 2.02 (s, 3H), 1.93 (s, 3H), 1.58 (m, 2H), 1.47 (m, 2H), 1.29
(m, 2H); δC(125 MHz; CDCl3) 173.2, 171.5, 170.6, 170.5 (m),
170.4 (m), 170.1, 170.0, 169.5, 168.8, 156.7, 101.0, 93.7, 78.2,
75.9, 74.7, 73.2, 72.5, 71.2, 71.0, 70.9, 70.8, 70.3, 69.9, 69.8,
67.9, 67.4, 67.0, 66.7, 64.9, 64.0, 62.1, 61.4, 61.3, 50.9, 40.9,
36.6, 29.8, 26.3, 24.8, 21.1, 21.0 (m), 20.9 (m), 20.8(4) (m),
20.7(8), 20.7(1); m/z (FAB MS) 1238 (Mϩ ϩ H. C51H76N5O30
requires m/z, 1238.46); 1276 (Mϩ ϩ K. C51H75N5O30ؒK requires
m/z, 1276.41).
Per-acetylated ꢀGal conjugate 21
From 18 (65 mg, 0.20 mmol), CDI (35 mg, 0.22 mmol), THF
(10 ml) and 14 (freshly prepared from 82 mg, 0.066 mmol of
13) was obtained compound 21 (66 mg, 65%) as a white solid;
δH(500 MHz; CDCl3) 7.98 (d, J = 8.5 Hz, 2H), 7.24 (m, 1H),
6.96 (d, J = 9.0 Hz, 2H), 6.48 (s, 1H), 6.21 (d, J = 10.0 Hz, 1H),
5.44 (m, 1H), 5.32–5.18 (m, 5H), 5.15 (dd, J = 10.2, 8.2 Hz, 1H),
5.08 (dd, J = 10.5, 3.0 Hz, 1H), 5.03 (m, 1H), 4.81 (t, J = 9.5 Hz,
1H), 4.39–4.36 (m, 2H), 4.19–4.01 (m, 10H), 3.82–3.62 (m,
16H), 3.13 (q, J = 6.7 Hz, 2H), 3.03 (t, J = 7.8 Hz, 2H), 2.63
(s, 3H), 2.22 (m, 2H), 2.15 (s, 3H), 2.12 (s, 6H), 2.08 (s, 3H), 2.06
(s, 3H), 2.05 (s, 3H), 2.04(5) (s, 3H), 2.03(6) (s, 3H), 2.02 (s, 3H),
1.94 (s, 3H), 1.58 (m, 2H), 1.47 (m, 2H), 1.28 (m, 2H); δC(125
MHz; CDCl3) 176.5, 174.4, 173.2, 171.5, 170.6, 170.5 (m), 170.4
(m), 170.1, 170.0, 169.5, 168.8, 168.2, 161.1, 159.4, 158.9, 156.7,
129.2, 119.8, 114.9, 101.3, 101.0, 93.7, 78.2, 75.9, 74.7, 73.2,
72.5, 71.2, 71.0, 70.7, 70.5, 69.9, 69.8 (m), 67.9, 67.4, 67.0, 66.7,
66.5, 64.9, 64.0, 62.2, 61.4, 61.3, 40.9, 39.5, 36.6, 29.8, 27.3,
26.4, 24.9, 23.7, 21.1, 21.0 (m), 20.9 (m), 20.8(5) (m), 20.7(9),
20.7(1), 12.6; m/z (FAB MS) 1561 (Mϩ ϩ K. C67H90N6O34ؒK
requires m/z, 1561.51).
Per-acetylated ꢀGal conjugate 19
From 18 (74 mg, 0.22 mmol), CDI (40 mg, 0.24 mmol), THF
(10 ml) and 9 (freshly prepared from 106 mg, 0.1 mmol of 8)
was obtained compound 19 (63 mg, 47%) as a white solid;
δH(500 MHz; CDCl3) 7.98 (d, J = 8.5 Hz, 2H), 6.96 (d, J = 9.0
Hz, 2H), 6.84 (t, J = 5.8 Hz, 1H), 6.49 (s, 1H), 6.16 (d, J = 9.0
Hz, 1H), 5.44 (m, 1H), 5.32–5.23 (m, 4H), 5.20 (t, J = 9.8 Hz,
1H), 5.15 (dd, J = 10.2, 8.2 Hz, 1H), 5.09 (dd, J = 10.8, 3.2 Hz,
1H), 4.81 (t, J = 9.5 Hz, 1H), 4.36 (m, 2H), 4.21–4.02 (m, 8H),
3.81 (dd, J = 10.0, 3.0 Hz, 1H), 3.78 (t, J = 6.8 Hz, 1H), 3.74 (m,
2H), 3.41 (q, J = 6.8 Hz, 2H), 3.04 (t, J = 7.5 Hz, 2H), 2.63 (s,
3H), 2.23 (m, 2H), 2.15 (s, 3H), 2.13 (s, 6H), 2.09 (s, 3H), 2.07 (s,
3H), 2.06 (s, 3H), 2.05 (s, 3H), 2.04 (s, 3H), 2.03 (s, 3H), 1.94 (s,
3H), 1.62 (m, 6H), 1.36 (m, 2H); δC(125 MHz; CDCl3) 176.6,
174.5, 173.1, 171.6, 170.6, 170.5 (m), 170.4 (m), 170.1, 170.0,
169.5, 168.9, 168.2, 161.1, 159.3, 158.9, 129.2, 119.8, 114.9,
101.3, 101.1, 93.7, 78.2, 75.9, 74.7, 73.2, 72.5, 71.2, 71.0, 69.8,
67.9, 67.4, 67.0, 66.7, 66.5, 64.9, 62.1, 61.4, 61.3, 39.3, 36.5,
29.3, 27.3, 26.4, 24.8, 23.7, 21.1, 20.9(9) (m), 20.9(5) (m), 20.8(5)
(m), 20.8, 20.7, 12.6; m/z (FAB MS) 1347.4 (Mϩ. C60H77N5O30
requires M, 1347.47); 1385.0 (Mϩ Ϫ H ϩ K. C60H76N5O30ؒK
requires m/z, 1385.42).
ꢀGal conjugate 1
To a solution of 19 (63 mg, 0.047 mmol) in absolute methanol
(20 ml) at 0 ЊC was added sodium methoxide to adjust pH to 9.
The mixture was stirred for 6 h and then neutralized with
Dowex 50WX2-100 (Hϩ) resin. The resin was filtered off, and
the filtrate was concentrated. Crystallization of the resultant
residue from boiling water afforded compound 1 (32 mg, 74%)
as a white solid; δH(500 MHz; CD3CN–D2O 1 : 1) 8.22 (d,
J = 8.5 Hz, 2H), 7.35 (d, J = 9.0 Hz, 2H), 6.81 (s, 1H), 5.35 (d,
J = 4.0 Hz, 1H), 5.18 (d, J = 9.0 Hz, 1H), 4.73 (d, J = 7.5 Hz,
1H), 4.43–4.37 (m, 4H), 4.22–4.20 (m, 1H), 4.16–4.12 (m, 2H),
4.08–3.85 (m, 11H), 3.66–3.58 (m, 4H), 3.31 (t, J = 7.8 Hz, 2H),
2.91 (s, 3H), 2.55 (t, J = 7.2 Hz, 2H), 2.49 (m, 2H), 2.29 (p,
J = 2.4 Hz, CH3CN), 1.88 (m, 4H), 1.64 (m, 2H); m/z (FAB MS)
950 (Mϩ ϩ Na. C40H57N5O20ؒNa requires m/z, 950.35).
ꢀGal conjugate 2
To a solution of 20 (65 mg, 0.045 mmol) in absolute methanol
(20 ml) at 0 ЊC was added sodium methoxide to adjust pH to 9.
The mixture was stirred for 6 h and then neutralized with
Dowex 50WX2-100 (Hϩ) resin. The resin was filtered off, and
the filtrate was concentrated. Crystallization of the resultant
residue from boiling water afforded compound 2 (34 mg, 73%)
as a slightly yellowish solid; δH(500 MHz; CD3CN–D2O 1 : 1)
8.18 (d, J = 9.0 Hz, 2H), 7.31 (d, J = 8.5 Hz, 2H), 6.78 (s, 1H),
5.32 (d, J = 3.5 Hz, 1H), 5.15 (d, J = 10.0 Hz, 1H), 4.70 (d,
J = 7.5 Hz, 1H), 4.40–4.33 (m, 4H), 4.19 (m, 1H), 4.13–4.09 (m,
2H), 4.06–3.82 (m, 11H), 3.63–3.57 (m, 4H), 3.35 (t, J = 7.0 Hz,
2H), 3.28 (t, J = 7.2 Hz, 2H), 2.88 (s, 3H), 2.50 (t, J = 7.0 Hz,
2H), 2.46 (t, J = 6.5 Hz, 2H), 2.42 (t, J = 7.8 Hz, 2H), 2.27
(p, J = 2.5 Hz, CH3CN), 1.82 (m, 6H), 1.69 (m, 2H), 1.55 (m,
4H); m/z (FAB MS) 1041 (Mϩ ϩ H. C46H69N6O21 requires
m/z, 1041.45); 1063 (Mϩ ϩ Na. C46H68N6O21ؒNa requires
m/z, 1063.43); 1079 (Mϩ ϩ K. C46H68N6O21ؒK requires m/z,
1079.41).
Per-acetylated ꢀGal conjugate 20
From 8 (106 mg, 0.1 mmol) following the sequence of hydro-
genation (to 9), acylation (to 10) and azide substitution was
obtained compound 11 (90 mg, 77%). From 18 (63 mg, 0.19
mmol), CDI (34 mg, 0.21 mmol), THF (10 ml) and 12 (freshly
prepared from 90 mg, 0.077 mmol of 11) was obtained com-
pound 20 (65 mg, 58%) as a white solid; δH(500 MHz; CDCl3)
7.98 (d, J = 8.5 Hz, 2H), 6.95 (d, J = 8.5 Hz, 2H), 6.91 (t, J = 5.8
Hz, 1H), 6.49 (s, 1H), 6.28 (d, J = 9.5 Hz, 1H), 5.65 (t, J = 5.8
Hz, 1H), 5.44 (m, 1H), 5.32–5.18 (m, 5H), 5.15 (dd, J = 10.2, 7.8
Hz, 1H), 5.08 (dd, J = 10.5, 3.5 Hz, 1H), 4.81 (t, J = 9.5 Hz,
1H), 4.39 (m, 2H), 4.20–4.01 (m, 8H), 3.81 (dd, J = 10.0, 3.0 Hz,
1H), 3.78 (t, J = 7.2 Hz, 1H), 3.74 (m, 2H), 3.42 (q, J = 6.8 Hz,
2H), 3.21 (q, J = 6.5 Hz, 2H), 3.03 (t, J = 7.8 Hz, 2H), 2.63 (s,
3H), 2.22 (m, 2H), 2.15 (s, 3H), 2.12(2) (s, 3H), 2.11(8) (s, 3H),
2.08 (s, 3H), 2.06 (s, 3H), 2.05 (s, 3H), 2.04(4) (s, 3H), 2.03(7) (s,
J. Chem. Soc., Perkin Trans. 1, 2001, 1716–1722
1721