Synthesis of divalent β-(1→6)-branched (1→3)-glucohexaose and trivalent β-(1→6)-branched (1→3)-glucotriose

Article abstract of DOI:10.1016/j.carres.2004.09.016

Hexaose, β-d-Glcp-(1→3)-[β-d-Glcp-(1→6)]-α-d- Glcp-(1→3)-β-d-Glcp-(1→3)-[β-d-Glcp-(1→6)] -β-d-Glcp, based dimers were synthesized by twofold glycosidation of the hexaosyl trichloroacetimidate with hexylene 1,6-diol, diethylene glycol and triethylene glycol, respectively. Meanwhile, a triose, β-1d-Glcp-(1→3) -[β-d-Glcp-(1→6)]-β-d-Glcp, based trimer was obtained by glycosidation of the triosyl trichloroacetimidate with a glycerol-derived triol scaffold.

Full text of DOI:10.1016/j.carres.2004.09.016

Carbohydrate
RESEARCH
Carbohydrate Research 339 (2004) 2761–2768
Synthesis of divalent b-(1!6)-branched (1!3)-glucohexaose
and trivalent b-(1!6)-branched (1!3)-glucotriose
Zicheng Wu and Fanzuo Kong*
Research Center for Eco-Environmental Sciences, Academia Sinica, PO Box 2871, Beijing 100085, China
Received 12 July 2004; accepted 22 September 2004
Abstract—Hexaose, b-D-Glcp-(1!3)-[b-D-Glcp-(1!6)]-a-D-Glcp-(1!3)-b-D-Glcp-(1!3)-[b-D-Glcp-(1!6)]-b-D-Glcp, based dimers
were synthesized by twofold glycosidation of the hexaosyl trichloroacetimidate with hexylene 1,6-diol, diethylene glycol and trieth-
ylene glycol, respectively. Meanwhile, a triose, b-1D-Glcp-(1!3)-[b-D-Glcp-(1!6)]-b-D-Glcp, based trimer was obtained by glycos-
idation of the triosyl trichloroacetimidate with a glycerol-derived triol scaffold.
ꢀ 2004 Elsevier Ltd. All rights reserved.
Keywords: Oligosaccharide; Dendrimer; Glucose
1. Introduction
specifically dendritic ligands, polymeric ligands con-
structed on either peptide or acrylamide backbones
and liposomes or other multivalent presentations cre-
ated by self assembly of amphiphilic carbohydrates.
We present herein the syntheses of the glucohexaose-
based dimers 6, 9 and 10 by glycosidation of the hexao-
syl trichloroacetimidate with hexylene 1,6-diol, diethyl-
ene glycol and triethylene glycol, respectively, and the
synthesis of glucotriose, b-D-Glcp-(1!3)-[b-D-Glcp-
(1!6)]-D-Glcp, based glycocluster 16 by glycosidation
of the triosyl trichloroacetimidate with a glycerol-
derived triol scaffold. Among these compounds, 6 con-
tained a relatively hydrophobic spacer, while 9 and 10
had relatively hydrophilic spacers. From the studies
on their activity, we can observe the influence of spac-
ers. Besides, since we have not, thus far, obtained a def-
inite conclusion regarding the size of the active
fragment, dendrimer 16 was synthesized for testing
whether the trisaccharide was the possible minimum
active moiety.
A glucohexaose, b-D-Glcp-(1!3)-[b-D-Glcp-(1!6)]-a-
D-Glcp-(1!3)-b-D-Glcp-(1!3)-[b-D-Glcp-(16)]-D-Glcp,1
has been proved to have good immunoregulating activ-
ity. In combination with the chemotherapeutic agent
cyclophosphamide (CPA), the glucohexaose at a dose
of 0.5–1mg/kg substantially increased the inhibition of
Sarcoma 180 for CPA, but decreased the toxicity caused
by CPA.1 A study on the stimulatory effect on mouse
spleen showed that the hexaose has similar or better
stimulatory effect compared to lentinan.2
It is well known3,4 that most saccharide ligands bind
to their protein receptors only weakly, seldom showing
association constants beyond 106 M. Clearly, the effec-
tive in vivo control of events mediated by protein–
carbohydrate binding requires significantly greater
affinity. Against this backdrop, the development of
tight-binding ligands for carbohydrate-binding proteins
continues apace throughout the carbohydrate chemistry
and biology community. A wide range of multivalent
saccharide ligands have been reported.5–8 Most fit into
a relatively small number of conceptual frameworks,
2. Results and discussion
As shown in Scheme 1, hexasaccharide donor 19 was cou-
pled with excess hexylene glycol gave monoglycosidated
*
Corresponding author. Tel.: +86 10 62936613; fax: +86 10 62923563;
e-mail: fzkong@mail.rcees.ac.cn
0008-6215/$ - see front matter ꢀ 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2004.09.016

2762
Z. Wu, F. Kong / Carbohydrate Research 339 (2004) 2761–2768
BzO
BzO
BzO
BzO
O
BzO
O
O
BzO
O
BzO
O
HO(CH2)6OH
BzO
AcO
AcO
O
AcO
BzO
BzO
BzO
O
O
AcO
O
a
O
OC(NH)CCl3
AcO
O
AcO
AcO
BzO
1
BzO
BzO
BzO
BzO
BzO
BzO
O
O
O
O
BzO
O
BzO
AcO
AcO
O
O(CH2)6OH
AcO
BzO
BzO
BzO
O
O
AcO
O
O
AcO
O
AcO
AcO
BzO
2
BzO
BzO
BzO
BzO
BzO
BzO
O
O
O
HO(CH2CH2O)nH
O
BzO
O
BzO
AcO
O(CH2CH2O)nH
AcO
1
O
AcO
BzO
O
O
a
3 n = 2
4 n = 3
AcO
O
O
O
BzO
AcO
AcO
AcO
BzO
BzO
n = 2,3
OR1
R1O
R1O
R1O
OR1
OR2
OR2
R1O
O
O
O
OR2
OR1
R1O
O
O
O
OR1
OR1
OR1
R1O
OR2
O
OR2
R1O
R1O
O
O
OR1
R1O
OR2
O
O
R2O
R2O
O(CH2)6O
OR2
OR1
O
O
R2O
R2O
O
O
O
1
+
2
OR1
R1O
R2O
O
O
a
O
O
OR1
OR1
R2O
R2O
R1O
O
O
OR1
R1O
5 R1 = Bz, R2 = Ac
6 R1 = R2 = H
b
OR1
R1O
OR1
OR2
OR2
R1O
R1O
R1O
O
R1O
O
OR2
OR1
O
O
R1O
O
O
OR1
R1O
OR2
OR1
OR1
OR2
O
R1O
O
O
OR1
OR2
OR2
O
O
R2O
R2O
O(CH2CH2O)n
O
1 + 3 or 4
O
R2O
R1O
R1O
R1O
O
a
R2O
OR1
OR1
OR1
O
O
OR1
R2O
O
O
O
O
OR1
R2O
R2O
O
O
R1O
7 n = 2 R1 = Bz, R2 = Ac
9 n = 2 R1 = R2 = H
8 n = 3 R1 = Bz, R2 = Ac
10 n = 3 R1 = R2 = H
b
b
Scheme 1. Reagents and conditions: (a) TMSOTf (0.01–0.05equiv), CH2Cl2, ꢀ20 to 0ꢁC, 2–4h; (b) satd NH3–MeOH, rt, a week.
compound 2 in good yield (89.7%). Subsequent conden-
sation of 2 with 1 afforded diglycosidated compound
5 in satisfactory yield (80.3%). A trial for completion
of the glycosidation with one coupling reaction, that
is, coupling of two equivalents hexaosyl donors with
one equivalent of hexylene glycol, was also carried out
giving hardly any separated product. Similarly, diethyl-
ene and triethylene glycol linked hexaose dimers were
also obtained with two steps. Thus, glycosidations of 1
with excess diethylene glycol and triethylene glycol were
successfully performed producing 3 (90.2%) and 4
(88.2%), respectively. Subsequent condensation of 3
and 4 with 1 furnished the dimers 7 (76.5%) and 8
(68.8%), respectively. Deacylation of 5, 7 and 8 in
ammonium-saturated methanol gave the dimers 6, 9
and 10, respectively.
For preparation of dendrimers, a core scaffold
trialcohol 11 (Scheme 2) was obtained from glycerol
by employing an iterative allylation–hydroboration
strategy in satisfactory overall yield.10 However, gly-
cosidation of 1 with 11 only produced a diglycosylated
mixture, and the triglycosylated cluster was not
obtainable even repeating the coupling–separation
process for five times. This indicated that clusters with
a large oligosaccharide as the ligand were difficult to
prepare if the scaffold did not have long enough spac-
ers. In contrast, a triose-based glycocluster 16 was suc-
cessfully obtained by repeated glycosylation of 11 with
excess triosyl donor 14 followed by deprotection.
Bioactivity evaluation of 6, 9, 10, 13 and 16 is in
progress, and the results will be reported in due
course.

Z. Wu, F. Kong / Carbohydrate Research 339 (2004) 2761–2768
2763
R2 = R, R3 = H
or
O
HO
HO
HO
O
O
RO
O
O
R2O
O
O
O
O
O
a
1
+
R3O
O
O
R
2 = H, R3 = R
12
11
BzO
BzO
BzO
BzO
BzO
BzO
O
O
O
O
BzO
O
BzO
AcO
R=
AcO
O
AcO
BzO
BzO
BzO
O
O
AcO
O
O
AcO
O
AcO
AcO
BzO
R2 = R, R3 = H
or
O
RO
R2O
R3O
O
O
O
13
O
O
b
R
2 = H, R3 = R
HO
HO
HO
HO
HO
HO
O
O
O
O
HO
O
HO
HO
HO
R=
O
HO
HO
HO
HO
O
O
HO
O
O
HO
O
HO
HO
HO
BzO
BzO
BzO
O
O
BzO
BzO
BzO
BzO
O
O
BzO
BzO
BzO
O
O
AcO
O
O
O
O
AcO
+
11
BzO
BzO
BzO
O
AcO
BzO
BzO
BzO
a
O
O
O
BzO
AcO
O
OC(NH)CCl3
BzO
BzO
BzO
BzO
O
O
O
O
O
O
AcO
O
O
BzO
14
AcO
BzO
BzO
BzO
O
BzO
O
BzO
BzO
O
O
AcO
BzO
O
O
BzO
AcO
BzO
BzO
15
HO
O
HO
O
HO
HO
HO
O
O
O
O
O
O
HO
O
HO
HO
HO
HO
HO
HO
b
O
HO
O
HO
HO
HO
O
O
O
HO
O
O
HO
HO
HO
HO
O
HO
O
HO
HO
HO
HO
HO
O
O
O
O
HO
O
O
HO
HO
16
Scheme 2. Reagents and conditions: (a) TMSOTf (0.01–0.05equiv), CH2Cl2, ꢀ20 to 0ꢁC, 2–4h; (b) satd NH3–MeOH, rt, a week.
3. Experimental
3.1. General methods
Perkin–Elmer model 241-MC automatic polarimeter
1
for solutions in a 1-dm, jacketed cell. H NMR and
13C NMR spectra were recorded with Varian XL-400
spectrometers, for solutions in CDCl3 or in D2O as
indicated. Chemical shifts are expressed in ppm down-
field from the internal Me4Si absorption. Mass spectra
Melting points were determined with a ÔMel-TempÕ
apparatus. Optical rotations were determined with a

2764
Z. Wu, F. Kong / Carbohydrate Research 339 (2004) 2761–2768
were measured using MALTI-TOF-MS with CCA as
matrix or recorded with a VG PLATFORM mass spec-
trometer using the ESI mode. Thin-layer chromatogra-
phy (TLC) was performed on silica gel HF with
detection by charring with 30% (v/v) sulfuric acid in
MeOH or by UV detection. Column chromatography
was conducted by elution of a column (8 · 100mm,
16 · 240mm, 18 · 300mm, 35 · 400mm) of silica gel
(100–200 mesh) with EtOAc–petroleum ether (bp 60–
90ꢁC) as the eluent. Analytical LC was performed with
a Gilson HPLC consisting of a pump (model 306),
stainless steel column packed with silica gel (Spherisorb
SiO2, 10 · 300mm or 4.6 · 250mm), differential refrac-
tometer (132-RI Detector), UV–vis detector (model
118). EtOAc–petroleum ether (bp 60–90ꢁC) was used
as the eluent at a flow rate of 1–4mL/min. Solutions
were concentrated at a temperature <60ꢁC under
diminished pressure.
2.03, 1.97, 1.84, 1.78, 1.62 (s, 21H, 7 CH3CO), 1.58–
1.52 (m, 8H, –CH2–); 13C NMR (CDCl3, 100MHz):
d 170.5, 170.1, 169.6, 169.5, 169.5, 169.3, 168.9 (7C, 7
COCH3), 166.1, 166.0, 166.0, 166.0, 165.9, 165.6,
165.6, 165.2, 165.2, 165.1, 165.0, 165.0 (12C, 12 COPh),
101.3, 101.3, 101.1, 100.6, 100.4 (5b-C-1), 93.4 (a-C-1);
Anal. Calcd for C140H136O51: C, 63.82; H, 5.20. Found:
C, 64.01; H, 5.12.
3.3. 2-(2-Hydroxyethoxy)ethyl 2,3,4,6-tri-O-benzoyl-b-D-
glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glu-
copyranosyl-(1!6)]-2,4-di-O-acetyl-a-D-glucopyranosyl-
(1!3)-2,4,6-tri-O-acetyl-b-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-b-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-b-D-glucopyranoside (3)
Compound 1 (1.00g, 0.373mmol) and diethylene glycol
(370mg, 3.49mmol) were dried together under high
vacuum for 2h, then dissolved in anhyd CH2Cl2
(20mL). TMSOTf (15lL, 0.132mmol) was added
dropwise at ꢀ20ꢁC with N2 protection. The reaction
mixture was stirred for 3h, during which time the tem-
perature was gradually raised to ambient temperature.
Then the mixture was neutralized with Et3N. Concen-
tration of the reaction mixture, followed by purifica-
tion on a silica gel column with 1:1 petroleum ether–
EtOAc as the eluent gave the product 3 (823mg,
90.2%) as a foamy solid: [a]D +40.8 (c 1.0, CHCl3);
1H NMR (CDCl3, 400MHz): d 8.05–7.26 (m, 60H,
12 Bz-H), 5.91 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4),
5.86 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4), 5.85 (dd, 1H,
3.2. 6-Hydroxyhexyl 2,3,4,6-tri-O-benzoyl-b-D-glucopy-
ranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glucopyrano-
syl-(1!6)]-2,4-di-O-acetyl-a-D-glucopyranosyl-(1!3)-
2,4,6-tri-O-acetyl-b-D-glucopyranosyl-(1!3)-[2,3,4,6-
tetra-O-benzoyl-b-D-glucopyranosyl-(1!6)]-2,4-di-O-
acetyl-b-D-glucopyranoside (2)
Compound 1 (1g, 0.373mmol) and 1,6-hexanediol
(400mg, 3.39mmol) were dried together under high
vacuum for 2h, then dissolved in anhyd CH2Cl2
(20mL). TMSOTf (15lL, 0.132mmol) was added
dropwise at ꢀ20ꢁC with N2 protection. The reaction
mixture was stirred for 3h, during which time the tem-
perature was gradually raised to ambient temperature.
Then the mixture was neutralized with Et3N. Concen-
tration of the reaction mixture, followed by purifica-
tion on a silica gel column with 1:1 petroleum ether–
EtOAc as the eluent gave the product 2 (881mg,
89.7%) as a foamy solid: [a]D +37.2 (c 1.0, CHCl3);
1H NMR (CDCl3, 400MHz): d 8.03–7.27 (m, 60H,
12 Bz-H), 5.88 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4),
5.86 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4), 5.83 (dd, 1H,
J3,4 = J4,5 = 9.7Hz, H-4), 5.76 (dd, 1H, J2,3 = J3,4
=
9.7Hz, H-3), 5.68 (dd, 1H, J2,3 = J3,4 =9.7Hz, H-3),
5.53 (dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.52 (dd, 1H,
J1,2 7.9Hz, J2,3 9.7Hz, H-2), 5.37 (dd, 1H, J1,2
7.9Hz, J2,3 9.7Hz, H-2), 5.35 (dd, 1H, J1,2 7.9Hz,
J2,3 9.7Hz, H-2), 4.98 (d, 1H, J1,2 7.9Hz, H-1), 4.93
(d, 1H, J1,2 7.9Hz, H-1), 4.91 (dd, 1H,
J3,4 = J4,5 = 9.7Hz, H-4), 4.90 (dd, 1H, J1,2 7.9Hz,
J2,3 9.7Hz, H-2), 4.87 (dd, 1H, J1,2 3.6Hz, J2,3
9.7Hz, H-2), 4.80 (d, 1H, J1,2 3.6Hz, H-1), 4.77 (d,
1H, J1,2 7.9Hz, H-1), 4.74–4.63 (m, 4H), 4.59 (d, 1H,
J1,2 7.9Hz, H-1), 4.56 (dd, 1H, J5,6 4.7Hz, J6,6
12.4Hz, H-6), 4.50–4.46 (m, 3H), 4.40 (dd, 1H, J5,6
4.7Hz, J6,6 12.4Hz, H-6), 4.32 (d, 1H, J1,2 7.9Hz, H-
1), 4.28 (dd, 1H, J5,6 4.7Hz, J6,6 12.4Hz, H-6), 4.20–
4.06 (m, 5H), 3.96–3.88 (m, 5H), 3.85–3.80 (m, 4H),
3.70–3.59 (m, 6H), 3.40–3.36 (m, 1H), 3.18–3.13 (m,
1H), 2.38, 2.05, 2.03, 1.97, 1.84, 1.79, 1.61 (s, 21H, 7
CH3CO); 13C NMR (CDCl3, 100MHz): d 170.6,
170.1, 169.6, 169.4, 169.4, 169.3, 169.2 (7C, 7 COCH3),
166.1, 166.0, 166.0, 166.0, 165.8, 165.8, 165.6, 165.6,
165.4, 165.2, 165.1, 165.0 (12C, 12 COPh), 101.3,
101.3, 101.1, 100.7, 100.4 (5b-C-1), 93.0 (a-C-1); Anal.
Calcd for C138H132O52: C, 63.21; H, 5.04. Found: C,
63.42; H, 4.99.
J3,4 = J4,5 = 9.7Hz, H-4), 5.76 (dd, 1H, J2,3
=
J3,4 = 9.7Hz, H-3), 5.67 (dd, 1H, J2,3 = J3,4 = 9.7Hz,
H-3), 5.54 (dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.52
(dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 5.36 (dd, 1H,
J1,2 7.9Hz, J2,3 9.7Hz, H-2), 5.35 (dd, 1H, J1,2
7.9Hz, J2,3 9.7Hz, H-2), 4.97 (d, 1H, J1,2 7.9Hz, H-
1), 4.94 (d, 1H, J1,2 7.9Hz, H-1), 4.91 (dd, 1 H,
J3,4 = J4,5 = 9.7Hz, H-4), 4.88 (dd, 1H, J1,2 7.9Hz,
J2,3 9.7Hz, H-2), 4.87 (dd, 1H, J1,2 3.6Hz, J2,3
9.7Hz, H-2), 4.79 (d, 1H, J1,2 3.6Hz, H-1), 4.75 (d,
1H, J1,2 7.9Hz, H-1), 4.71–4.64 (m, 4H), 4.59 (d, 1H,
J1,2 7.9Hz, H-1), 4.50–4.46 (m, 3H), 4.30 (dd, 1H,
J5,6 4.7Hz, J6,6 12.4Hz, H-6), 4.21 (d, 1H, J1,2
7.9Hz, H-1), 4.19–4.06 (m, 5H), 4.01–3.82 (m, 7H),
3.73–3.51 (m, 4H), 3.38–3.34 (m, 4H), 2.38, 2.05,

Z. Wu, F. Kong / Carbohydrate Research 339 (2004) 2761–2768
2765
3.4. 2-[2-(2-Hydroxyethoxy)ethoxy]ethyl 2,3,4,6-tri-O-
benzoyl-b-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-ben-
zoyl-b-D-glucopyranosyl-(1!6)]-2,4-di-O-acetyl-a-D-glu-
copyranosyl-(1!3)-2,4,6-tri-O-acetyl-b-D-glucopyrano-
syl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glucopyranosyl-
(1!6)]-2,4-di-O-acetyl-b-D-glucopyranoside (4)
TMSOTf (20lL, 0.176mmol) was added dropwise at
ꢀ20ꢁC with N2 protection. The reaction mixture was
stirred for 3h, during which time the temperature was
gradually raised to ambient temperature. Then the mix-
ture was neutralized with Et3N. Concentration of the
reaction mixture, followed by purification on a silica
gel column with 1:1.5 petroleum ether–EtOAc as the
eluent gave the product 5 (786mg, 80.3%) as a foamy
solid: [a]D +63.2 (c 1.0, CHCl3); 1H NMR (CDCl3,
400MHz): d 8.01–7.25 (m, 60H, 12 Bz-H), 6.11 (dd,
Compound 1 (1.00g, 0.373mmol) and triethylene glycol
(500mg, 3.33mmol) were dried together under high
vacuum for 2h, then dissolved in anhyd CH2Cl2
(20mL). TMSOTf (15lL, 0.132mmol) was added
dropwise at ꢀ20ꢁC with N2 protection. The reaction
mixture was stirred for 3h, during which time the
temperature was gradually raised to ambient tempera-
ture. Then the mixture was neutralized with Et3N. Con-
centration of the reaction mixture, followed by
purification on a silica gel column with 1:1 petroleum
1H, J3,4 = J4,5 = 9.7Hz, H-4), 5.87 (dd, 1H, J3,4
=
J4,5 = 9.7Hz, H-4), 5.85 (dd, 1H, J3,4 = J4,5 = 9.7Hz,
H-4), 5.74 (dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.70 (dd,
1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.56 (dd, 1H,
J2,3 = J3,4 = 9.7Hz, H-3), 5.53 (dd, 1H, J1,2 7.9Hz, J2,3
9.7Hz, H-2), 5.37 (dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-
2), 5.32 (dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 4.99 (d,
1H, J1,2 7.9Hz, H-1), 4.95 (d, 1H, J1,2 7.9Hz, H-1),
4.90 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4), 4.88 (dd, 1H,
J1,2 7.9Hz, J2,3 9.7Hz, H-2), 4.86 (dd, 1H, J1,2 3.6Hz,
J2,3 9.7Hz, H-2), 4.80 (d, 1H, J1,2 3.6Hz, H-1), 4.76
(d, 1H, J1,2 7.9Hz, H-1), 4.71–4.64 (m, 4H), 4.57 (d,
1H, J1,2 7.9Hz, H-1), 4.49–4.38 (m, 5H), 4.30 (dd, 1H,
J5,6 4.7Hz, J6,6 12.4Hz, H-6), 4.26 (d, 1H, J1,2 7.9Hz,
H-1), 4.19–4.06 (m, 6H), 4.01–3.82 (m, 5H), 3.78–3.51
(m, 4H), 3.39–3.32 (m, 3H), 2.32, 2.04, 2.02, 1.96,
1.83, 1.75, 1.62 (s, 21H, 7 CH3CO), 1.32–1.25 (m, 4H,
–CH2–); 13C NMR (CDCl3, 100MHz): d 170.6, 170.1,
169.7, 169.6, 169.6, 169.3, 169.3 (7C, 7 COCH3),
166.0, 166.0, 165.9, 165.9, 165.8, 165.6, 165.2, 165.2,
165.2, 165.0, 165.0, 165.0 (12C, 12 COPh), 101.4,
101.3, 101.0, 100.1, 100.4 (5b-C-1), 93.6 (a-C-1); ESIMS
for C274H258O100 (5152): 5173 [M+Na]+. Anal. Calcd
for C274H258O100: C, 63.89; H, 5.05. Found: C, 64.08;
H, 5.12.
ether–EtOAc as the eluent gave the product
2
(877mg, 88.2%) as a foamy solid: [a]D +43.9 (c 1.0,
1
CHCl3); H NMR (CDCl3, 400MHz): d 8.03–7.26 (m,
60H, 12 Bz-H), 5.90 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-
4), 5.86 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4), 5.84 (dd,
1H, J3,4 = J4,5 = 9.7Hz, H-4), 5.75 (dd, 1H, J2,3
=
J3,4 = 9.7Hz, H-3), 5.67 (dd, 1H, J2,3 = J3,4 = 9.7Hz,
H-3), 5.53 (dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.51 (dd,
1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 5.37 (dd, 1H, J1,2
7.9Hz, J2,3 9.7Hz, H-2), 5.35 (dd, 1H, J1,2 7.9Hz, J2,3
9.7Hz, H-2), 4.97 (d, 1H, J1,2 7.9Hz, H-1), 4.93 (d,
1H, J1,2 7.9Hz, H-1), 4.91 (dd, 1H, J3,4
=
J4,5 = 9.7Hz, H-4), 4.90 (dd, 1H, J1,2 7.9Hz, J2,3
9.7Hz, H-2), 4.86 (dd, 1H, J1,2 3.6Hz, J2,3 9.7Hz, H-
2), 4.80 (d, 1H, J1,2 3.6Hz, H-1), 4.75 (d, 1H, J1,2
7.9Hz, H-1), 4.73–4.60 (m, 4H), 4.59 (d, 1H, J1,2
7.9Hz, H-1), 4.57 (dd, 1H, J5,6 4.7Hz, J6,6 12.4Hz,
H-6), 4.50–4.45 (m, 3H), 4.33 (d, 1H, J1,2 7.9Hz, H-
1), 4.28 (dd, 1H, J5,6 4.7Hz, J6,6 12.4Hz, H-6), 4.21–
4.08 (m, 5H), 4.02–3.79 (m, 5H), 3.71–3.58 (m, 7H),
3.55–3.54 (m, 6H), 3.43–3.29 (m, 4H), 2.36, 2.04, 2.02,
1.96, 1.84, 1.77, 1.63 (s, 21H, 7 CH3CO); 13C NMR
(CDCl3, 100MHz): d 170.5, 170.1, 169.6, 169.5, 169.5,
169.4, 169.1 (7 C, 7 COCH3), 166.1, 166.0, 166.0,
165.8, 165.8, 165.6, 165.6, 165.3, 165.2, 165.1, 165.0,
164.9 (12C, 12 COPh), 101.4, 101.3, 101.3, 100.6, 100.3
(5b-C-1), 93.1 (a-C-1); Anal. Calcd for C140H136O53:
C, 63.06; H, 5.11. Found: C, 63.25; H, 5.06.
3.6. Hexyl-1,6-diyl bis{b-D-glucopyranosyl-(1!3)-[b-D-
glucopyranosyl-(1!6)]-a-D-glucopyranosyl-(1!3)-b-D-
glucopyranosyl-(1!3)-[b-D-glucopyranosyl-(1!6)]-b-D-
glucopyranoside} (6)
Compound 5 (500mg, 0.097mmol) was dissolved in a
satd solution of NH3 in MeOH (15mL). After a week
at rt, the reaction mixture was concentrated and the res-
idue was purified by chromatography on Sephadex LH-
20 (MeOH) to afford 6 (174mg, 87.2%) as a foamy solid:
1
3.5. Hexyl-1,6-diyl bis{2,3,4,6-tri-O-benzoyl-b-D-glu-
copyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glucopy-
ranosyl-(1!6)]-2,4-di-O-acetyl-a-D-glucopyranosyl-
(1!3)-2,4,6-tri-O-acetyl-b-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-b-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-b-D-glucopyranoside} (5)
[a]D +14.6 (c 1.0, H2O); H NMR (D2O, 400MHz): d
5.20 (d, 1H, J 3.2Hz, H-1), 4.66 (d, 1H, J 8.0Hz, H-
1), 4.64 (d, 1H, J 8.0Hz, H-1), 4.40 (d, 1H, J 8.0Hz,
H-1), 4.38 (d, 1H, J 8.0Hz, H-1), 4.36 (d, 1H, J
8.0Hz, H-1), 4.12–4.00 (m, 4H), 3.83–3.78 (m, 8H),
3.62–3.53 (m, 11H), 3.40–3.15 (m, 17H), 1.58–1.52 (m,
4H, –CH2–); 13C NMR (D2O, 100MHz): d 102.8,
102.8, 102.7, 102.7, 101.9 (5b-C-1), 99.1 (a-C-1). Anal.
Calcd for C78H134O62: C, 45.39; H, 6.50. Found: C,
45.54 H, 6.44.
Compound 1 (610mg, 0.227mmol) and 2 (500mg,
0.190mmol) were dried together under high vacuum
for 2h, then dissolved in anhyd CH2Cl2 (30mL).

2766
Z. Wu, F. Kong / Carbohydrate Research 339 (2004) 2761–2768
3.7. 3-Oxapent-1,5-diyl bis{2,3,4,6-tri-O-benzoyl-b-D-
glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glu-
copyranosyl-(1!6)]-2,4-di-O-acetyl-a-D-glucopyranosyl-
(1!3)-2,4,6-tri-O-acetyl-b-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-b-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-b-D-glucopyranoside} (7)
TMSOTf (30lL, 0.264mmol) was added dropwise at
ꢀ20ꢁC with N2 protection. The reaction mixture was
stirred for 3h, during which time the temperature was
gradually raised to ambient temperature. Then the mix-
ture was neutralized with Et3N. Concentration of the
reaction mixture, followed by purification on a silica
gel column with 1:1.5 petroleum ether–EtOAc as the
eluent gave the product 8 (670mg, 68.8%) as a foamy
solid: [a]D +42.3 (c 1.0, CHCl3); 1H NMR (CDCl3,
400MHz): d 7.95–7.24 (m, 60H, 12 Bz-H), 5.93 (dd, 1
Compound 1 (613mg, 0.229mmol) and 3 (500mg,
0.191mmol) were dried together under high vacuum
for 2h, then dissolved in anhyd CH2Cl2 (30mL).
TMSOTf (25lL, 0.220mmol) was added dropwise at
ꢀ20ꢁC with N2 protection. The reaction mixture was
stirred for 3h, during which time the temperature was
gradually raised to ambient temperature. Then the mix-
ture was neutralized with Et3N. Concentration of the
reaction mixture, followed by purification on a silica
gel column with 1:1.5 petroleum ether–EtOAc as the elu-
ent gave the product 7 (751mg, 76.5%) as a foamy solid:
[a]D +35.1 (c 1.0, CHCl3); 1H NMR (CDCl3, 400MHz):
H, J3,4 = J4,5 = 9.7Hz, H-4), 5.87 (dd, 1H, J3,4
=
J4,5 = 9.7Hz, H-4), 5.84 (dd, 1H, J3,4 = J4,5 = 9.7Hz,
H-4), 5.74 (dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.67
(dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.56 (dd, 1H,
J2,3 = J3,4 = 9.7Hz, H-3), 5.53 (dd, 1H, J1,2 7.9Hz,
J2,3 9.7Hz, H-2), 5.37 (dd, 1H, J1,2 7.9Hz, J2,3
9.7Hz, H-2), 5.35 (dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-
2), 4.96 (d, 1H, J1,2 7.9Hz, H-1), 4.93 (d, 1H, J1,2
7.9Hz, H-1), 4.90 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4),
4.88 (dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 4.86
(dd, 1H, J1,2 3.6Hz, J2,3 9.7Hz, H-2), 4.81 (d, 1H,
J1,2 3.6Hz, H-1), 4.76 (d, 1H, J1,2 7.9Hz, H-1),
4.72–4.55 (m, 5H), 4.54 (d, 1H, J1,2 7.9Hz, H-
1), 4.50–4.45 (m, 3H), 4. 29 (d, 1H, J1,2 7.9Hz, H-1),
4.26 (dd, 1H, J5,6 4.7Hz, J6,6 12.4Hz, H-6), 4.19–4.09
(m, 5H), 4.02–3.84 (m, 6H), 3.78–3.72 (m, 4H),
3.68–3.50 (m, 6H), 3.42–3.35 (m, 4H), 2.34, 2.05,
2.02, 1.96, 1.83, 1.77, 1.66 (s, 21H, 7 CH3CO); 13C
d 8.01–7.25 (m, 60H, 12 Bz-H), 5.97 (dd, 1H, J3,4
=
J4,5 = 9.7Hz, H-4), 5.86 (dd, 1H, J3,4 = J4,5 = 9.7Hz,
H-4), 5.85 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4), 5.73 (dd,
1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.70 (dd, 1H, J2,3
=
J3,4 = 9.7Hz, H-3), 5.57 (dd, 1H, J2,3 = J3,4 = 9.7Hz,
H-3), 5.55 (dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 5.38
(dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 5.34 (dd, 1H,
J1,2 7.9Hz, J2,3 9.7Hz, H-2), 4.93 (d, 1H, J1,2 7.9Hz,
H-1), 4.92 (d, 1H, J1,2 7.9Hz, H-1), 4.90 (dd, 1H,
J3,4 = J4,5 = 9.7Hz, H-4), 4.88 (dd, 1H, J1,2 7.9Hz, J2,3
9.7Hz, H-2), 4.86 (dd, 1H, J1,2 3.6Hz, J2,3 9.7Hz, H-
2), 4.81 (d, 1H, J1,2 3.6Hz, H-1), 4.78 (d, 1H, J1,2
7.9Hz, H-1), 4.76–4.68 (m, 4H), 4.58 (d, 1H, J1,2
7.9Hz, H-1), 4.56 (dd, 1H, J5,6 4.7Hz, J6,6 12.4Hz, H-
6), 4.50–4.46 (m, 4H), 4.32 (d, 1H, J1,2 7.9Hz, H-1),
4.28 (dd, 1H, J5,6 4.7Hz, J6,6 12.4Hz, H-6), 4.19–4.09
(m, 5H), 3.96–3.75 (m, 6H), 3.85–3.80 (m, 4H), 3.70–
3.59 (m, 5H), 3.38–3.30 (m, 5H), 2.30, 2.05, 2.03, 1.98,
1.96, 1.83, 1.76 (s, 21H, 7 CH3CO); 13C NMR (CDCl3,
100MHz): d 170.6, 170.1, 169.6, 169.6, 169.4, 169.4,
168.8 (7C, 7 COCH3), 166.1, 166.0, 166.0, 165.9,
165.7, 165.6, 165.5, 165.2, 165.2, 165.2, 165.0, 165.0
(12C, 12 COPh), 101.4, 101.2, 101.0, 100.0, 100.0 (5b-
C-1), 93.2 (a-C-1); ESIMS for C272H254O101 (5141):
5163 [M+Na]+. Anal. Calcd for C272H254O101: C,
63.58; H, 4.95. Found: C, 63.31; H, 5.04.
NMR (CDCl3, 100MHz):
d 170.5, 170.1, 169.6,
169.5, 169.5, 169.4, 168.9 (7C, 7 COCH3), 166.1,
166.0, 166.0, 165.8, 165.7, 165.6, 165.5, 165.2, 165.2,
165.1, 165.0, 165.0 (12C, 12 COPh), 101.3, 101.2,
101.1, 100.6, 100.4 (5b-C-1), 93.4 (a-C-1); ESIMS for
C
C
5.10.
274H258O102 (5184): 5206 [M+Na]+. Anal. Calcd for
274H258O102: C, 63.50; H, 5.02. Found: C, 63.32; H,
3.9. 3-Oxapent-1,5-diyl bis{b-D-glucopyranosyl-(1!3)-
[b-D-glucopyranosyl-(1!6)]-a-D-glucopyranosyl-(1!3)-
b-D-glucopyranosyl-(1!3)-[b-D-glucopyranosyl-(1!6)]-
b-D-glucopyranoside} (9)
Compound 7 (500mg, 0.097mmol) was dissolved in a
satd solution of NH3 in MeOH (20mL). After a week
at rt, the reaction mixture was concentrated, and the res-
idue was purified by chromatography on Sephadex LH-
20 (MeOH) to afford 9 (175mg, 88.2%) as a foamy solid:
3.8. 3,6-Dioxaoct-1,8-diyl bis{2,3,4,6-tri-O-benzoyl-b-D-
glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glu-
copyranosyl-(1!6)]-2,4-di-O-acetyl-a-D-glucopyranosyl-
(1!3)-2,4,6-tri-O-acetyl-b-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-b-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-b-D-glucopyranoside} (8)
1
[a]D +32.7 (c 1.0, H2O); H NMR (D2O, 400MHz): d
5.20 (d, 1H, J 3.2Hz, H-1), 4.62 (d, 1H, J 8.0Hz, H-
1), 4.60 (d, 1H, J 8.0Hz, H-1), 4.41 (d, 1H, J 8.0Hz,
H-1), 4.39 (d, 1H, J 8.0Hz, H-1), 4.36 (d, 1H, J
8.0Hz, H-1), 4.12–3.98 (m, 5H), 3.92–3.78 (m, 8H),
3.65–3.51 (m, 14H), 3.40–3.15 (m, 17H); 13C NMR
(D2O, 100MHz): d 102.9, 102.8, 102.7, 102.7, 102.1
(5b-C-1), 99.1 (a-C-1). Anal. Calcd for C76H130O63: C,
44.49; H, 6.34. Found: C, 44.65; H, 6.28.
Compound 1 (600mg, 0.224mmol) and 4 (500mg,
0.188mmol) were dried together under high vacuum
for 2h, then dissolved in anhyd CH2Cl2 (30mL).

Z. Wu, F. Kong / Carbohydrate Research 339 (2004) 2761–2768
2767
3.10. 3,6-Dioxaoct-1,8-diyl bis{b-D-glucopyranosyl-
(1!3)-[b-D-glucopyranosyl-(1!6)]-a-D-glucopyranosyl-
(1!3)-b-D-glucopyranosyl-(1!3)-[b-D-glucopyranosyl-
(1!6)]-b-D-glucopyranoside} (10)
for 2h, then dissolved in anhyd CH2Cl2 (50mL).
TMSOTf (50lL, 0.440mmol) was added dropwise at
ꢀ20ꢁC with N2 protection. The reaction mixture was
stirred for 3h, during which time the temperature was
gradually raised to ambient temperature. Then the mix-
ture was neutralized with Et3N. Concentration of the
reaction mixture, followed by purification on a silica
gel column with 1:2 petroleum ether–EtOAc as the elu-
ent gave a product (500mg). With the product as the
starting acceptor and 1 as the donor, reiteration of the
coupling–separation process for four times produced
the mixture 12 (400mg, 40.2%): [a]D +45.2 (c 1.0,
Compound 8 (500mg, 0.096mmol) was dissolved in a
satd solution of NH3 in MeOH (20mL). After a week
at rt, the reaction mixture was concentrated, and the res-
idue was purified by chromatography on Sephadex LH-
20 (MeOH) to afford 10 (174mg, 84.6%) as a foamy so-
1
lid: [a]D +29.6 (c 1.0, H2O); H NMR (D2O, 400MHz):
d 5.20 (d, 1H, J 3.2Hz, H-1), 4.66 (d, 1H, J 8.0Hz, H-1),
4.62 (d, 1H, J 8.0Hz, H-1), 4.39 (d, 1H, J 8.0Hz, H-1),
4.38 (d, 1H, J 8.0Hz, H-1), 4.37 (d, 1H, J 8.0Hz, H-
1), 4.16–3.98 (m, 5H), 3.82–3.72 (m, 10H), 3.63–3.49
(m, 17H), 3.45–3.15 (m, 16H); 13C NMR (D2O,
100MHz): d 102.9, 102.8, 102.8, 102.7, 102.1 (5b-C-1),
99.1 (a-C-1). Anal. Calcd for C78H134O64: C, 44.70; H,
6.40. Found: C, 45.56 H, 6.33.
1
CHCl3); H NMR (CDCl3, 400MHz): d 8.02–7.26 (m,
60H, 12 Bz-H), 5.88 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4),
5.86 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-4), 5.85 (dd, 1H,
=
J3,4 = J4,5 = 9.7Hz, H-4), 5.74 (dd, 1H, J2,3
J3,4 = 9.7Hz, H-3), 5.67 (dd, 1H, J2,3 = J3,4 = 9.7Hz,
H-3), 5.60 (dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.54 (dd,
1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 5.37 (dd, 1H, J1,2
7.9Hz, J2,3 9.7Hz, H-2), 5.35 (dd, 1H, J1,2 7.9Hz, J2,3
9.7Hz, H-2), 4.93 (d, 1H, J1,2 7.9Hz, H-1), 4.90 (d,
1H, J1,2 7.9Hz, H-1), 4.86 (dd, 1H, J3,4 = J4,5 = 9.7Hz,
H-4), 4.85 (dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz, H-2), 4.82
(dd, 1H, J1,2 3.6Hz, J2,3 9.7Hz, H-2), 4.81 (d, 1H, J1,2
3.6Hz, H-1), 4.77 (d, 1H, J1,2 7.9Hz, H-1), 4.73–4.56
(m, 5H), 4.55 (d, 1H, J1,2 7.9Hz, H-1), 4.50–4.42 (m,
4H), 4.30 (d, 1H, J1,2 7.9Hz, H-1), 4.27 (dd, 1H, J5,6
4.7Hz, J6,6 12.4Hz, H-6), 4.19–4.09 (m, 4H), 4.04–3.81
(m, 6H), 3.78–3.59 (m, 7H), 3.68–3.50 (m, 6H), 3.42–
3.35 (m, 5H), 2.05, 2.02, 1.99, 1.96, 1.84, 1.79, 1.68 (s,
21H, 7 CH3CO), 1.66–1.46 (m, 4H); 13C NMR (CDCl3,
100MHz): d 170.5, 170.1, 169.6, 169.5, 169.4, 169.4,
168.7 (7 C, 7 COCH3), 166.1, 166.0, 165.8, 165.7,
165.6, 165.6, 165.2, 165.2, 165.2, 165.0, 165.0, 165.0
(12C, 12 COPh), 101.8, 101.4, 101.1, 100.6, 99.5 (5b-C-
1), 93.4 (a-C-1); ESIMS for C289H288O107 (5471): 5492
[M+Na]+. Anal. Calcd for C289H288O107: C, 63.42; H,
5.27. Found: C, 63.22; H, 5.20.
3.11. 1,2,3-Tris-O-[2-(2-Hydroxyethoxy)ethyl]glycerol (11)
To a solution of glycerol (5.0g, 54.3mmol) in dry DMF
(50mL), AllBr (17.3mL, 196mmol) and NaH (13.9g,
49% in oil, 285mmol) were added under cooling with
an ice bath. The mixture was stirred for 2h at rt, at
the end of which time TLC (3:1 petroleum ether–
EtOAc) indicated that the reaction was complete. The
mixture was poured to water and extracted with
CH2Cl2. The organic phase was concentrated, and the
residue was purified on a silica gel column with 1:1.5
petroleum ether–EtOAc as the eluent to give a syrupy
product (10.6g, 92.5%). Then the product (500mg,
2.36mmol) was dissolved in dry THF (20mL) and stir-
red. A solution of 9-BBN in THF (0.5M, 28.4mL,
14.2mmol) was added dropwise. The reaction mixture
was heated under reflux for 2h, and excess of 9-BBN
was then destroyed by dropwise addition of water at
0ꢁC. The hydroboration mixture was oxidized by treat-
ment with 3M aq NaOH (28.4mL) and 30% H2O2 solu-
tion (28.4mL) at 0ꢁC, followed by stirring overnight at
rt. The mixture was saturated with K2CO3 and extracted
with THF. The organic phase was concentrated, and the
residue was purified by a silica gel column with 1:3
petroleum ether–EtOAc to yield a trialcohol (511mg,
82.3%). The allylation–hydroboration process was re-
peated on the trialcohol to give compound 11 (600mg,
3.13. Diglycosylated mixture 13
Compound 12 (300mg, 0.055mmol) was dissolved in a
satd solution of NH3 in MeOH (15mL). After a week
at rt, the reaction mixture was concentrated, and the
residue was purified by chromatography on Sephadex
LH-20 (MeOH) to afford 13 (111mg, 84.6%) as a foamy
solid: [a]D +29.6 (c 1.0, H2O); 1H NMR (D2O,
400MHz): d 5.26 (d, 1H, J 3.2Hz, H-1), 4.67 (d, 1H, J
8.0Hz, H-1), 4.65 (d, 1H, J 8.0Hz, H-1), 4.48 (d, 1H,
J 8.0Hz, H-1), 4.46 (d, 1H, J 8.0Hz, H-1), 4.43 (d,
1H, J 8.0Hz, H-1), 4.16–4.02 (m, 5H), 3.88–3.82 (m,
10H), 3.80–3.52 (m, 16H), 3.52–3.12 (m, 23H), 1.93–
1.62 (m, 4H); 13C NMR (D2O, 100MHz): d 102.8,
102.8, 102.8, 102.7, 102.7 (5b-C-1), 99.1 (a-C-1). Anal.
Calcd for C93H164O69: C, 46.81; H, 6.88. Found: C,
47.04 H, 6.76.
1
53.4% overall): H NMR (CDCl3, 400MHz): d 3.83–
3.79 (m, 6H), 1.90–1.85 (m, 16H), 1.67–1.55 (m, 13H),
1.49–1.44 (m, 6H). Anal. Calcd for C21H44O9: C,
57.27; H, 10.00. Found: C, 57.46; H, 10.06.
3.12. Diglycosylated mixture 12
Compound 1 (2.51g, 0.940mmol) and 11 (80mg,
0.182mmol) were dried together under high vacuum

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Z. Wu, F. Kong / Carbohydrate Research 339 (2004) 2761–2768
3.14. Triose-based glycocluster 15
at rt, the reaction mixture was concentrated, and the res-
idue was purified by chromatography on Sephadex LH-
Compound 14 (1.18g, 0.752mmol) and 11 (50mg,
0.188mmol) were dried together under high vacuum
for 2h, then dissolved in anhyd CH2Cl2 (30mL).
TMSOTf (50lL, 0.440mmol) was added dropwise at
ꢀ20ꢁC with N2 protection. The reaction mixture was
stirred for 3h, during which time the temperature was
gradually raised to ambient temperature. Then the mix-
ture was neutralized with Et3N. Concentration of the
reaction mixture, followed by purification on a silica
gel column with 1:1.5 petroleum ether–EtOAc as the elu-
ent gave a product (300mg). Mass spectrometry of the
product indicated incomplete glycosylation. Thus, with
the product as the starting acceptor, reiteration of the
coupling–separation process for three times produced
the product 15 (456mg, 52.2%) as a foamy solid: [a]D
20 (MeOH) to afford 16 (126mg, 82.6%) as a foamy so-
1
lid: [a]D +36.2 (c 1.0, H2O); H NMR (D2O, 400MHz):
d 4.62 (d, 1H, J 8.0Hz, H-1), 4.50 (d, 1H, J 8.0Hz, H-1),
4.46 (d, 1H, J 8.0Hz, H-1), 4.15–4.04 (m, 4H), 3.92–3.76
(m, 5H), 3.62–3.55 (m, 6H), 3.46–3.20 (m, 12H) 1.93–
1.61 (m, 4H); 13C NMR (D2O, 100MHz): d 101.8,
100.5, 97.0 (3b-C-1). Anal. Calcd for C75H134O54: C,
47.67; H, 7.15. Found: C, 47.84 H, 7.31.
Acknowledgements
This work was supported by The Chinese Academy of
Sciences (KZCX3-J-08) and by The National Natural
Science Foundation of China (Projects 30070185 and
39970864).
1
+35.3 (c 1.0, CHCl3); H NMR (CDCl3, 400MHz): d
8.17–7.26 (m, 40H, 8 Bz-H), 5.88 (dd, 1H, J3,4
=
J4,5 = 9.7Hz, H-4), 5.85 (dd, 1H, J3,4 = J4,5 = 9.7Hz,
H-4), 5.68 (dd, 1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.65 (dd,
1H, J2,3 = J3,4 = 9.7Hz, H-3), 5.49 (dd, 1H, J1,2 7.9Hz,
J2,3 9.7Hz, H-2), 5.37 (dd, 1H, J1,2 7.9Hz, J2,3 9.7Hz,
H-2), 5.00 (d, 1H, J1,2 7.9Hz, H-1), 4.97 (d, 1H,
J = 7.9Hz, H-1), 4.85 (dd, 1H, J3,4 = J4,5 = 9.7Hz, H-
4), 4.74 (d, 1H, J = 7.9Hz, H-1), 4.67–4.60 (m, 3H),
4.52–4.41 (m, 3H), 4.19–4.05 (m, 6H), 4.01–3.83 (m,
4H), 3.48–3.22 (m, 4H) 2.04, 1.89 (s, 6H, 2 CH3CO)
1.69–1.51 (m, 4H); 13C NMR (CDCl3, 100MHz): d
170.4, 169.6 (2C, 2 COCH3), 166.1, 165.7, 165.6,
165.6, 165.2, 165.1, 165.0, 165.0 (8C, 8 COPh), 101.8,
101.4, 100.6, (3b-C-1); ESIMS for C255H242O84 (4649):
4670 [M+Na]+. Anal. Calcd for C245H242O84: C, 64.96
H, 5.35. Found: C, 64.72 H, 5.45.
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Compound 15 (400mg, 0.086mmol) was dissolved in a
satd solution of NH3 in MeOH (15mL). After a week