Synthesis of β-(1→6)-branched β-(1→3) glucohexaose and its analogues containing an α-(1→3) linked bond with antitumor activity

Article abstract of DOI:10.1016/S0968-0896(03)00118-4

A β-(1→6)-branched β-(1→3)-glucohexaose, present in many biologically active polysaccharides from traditionally herbal medicines such as Ganoderma lucidum, Schizophyllum commune and Lentinus edodes, was synthesized as its lauryl glycoside 32, and its anal

Full text of DOI:10.1016/S0968-0896(03)00118-4

Bioorganic & Medicinal Chemistry 11 (2003) 2193–2203
Synthesis of ꢀ-(1!6)-Branched ꢀ-(1!3) Glucohexaose and
Its Analogues Containing an ꢁ-(1!3) Linked Bond with
Antitumor Activity
Jun Ning,a,* Wenhui Zhang,a Yuetao Yi,a Guangbin Yang,a
Zhikui Wu,b Jie Yib and Fanzuo Konga,*
aResearch Center for Eco-Environmental Sciences, Chinese Academy of Sciences, PO Box 2871, Beijing 100085, PR China
bGuang Anmen Hospitol, Chinese Academy of Traditional Chinese Medicine, PR China
Received 24 December 2002; accepted 13 February 2003
Abstract—A b-(1!6)-branched b-(1!3)-glucohexaose, present in many biologically active polysaccharides from traditionally her-
bal medicines such as Ganoderma lucidum, Schizophyllum commune and Lentinus edodes, was synthesized as its lauryl glycoside 32,
and its analogues 18, 20 and 33 containing an a-(1!3) linked bond were synthesized. It is interesting to find that coupling of a 3,6-
branched acylated trisaccharide trichloroacetimidate donor 9 with 3,6-branched acceptors 13 and 16 with 30-OH gave the a-(1! 3)-
linked hexasaccharides 17 and 19, respectively, in spite of the presence of C-2 ester capable of neighboring group participation.
However, coupling of 9 with 4-methoxyphenyl 4,6-O-benzylidene-b-d-glucopyranoside (27) selectively gave b-(1!3)-linked tetra-
saccharide 28. Simple chemical transformation of the tetrasaccharide 28 gave acylated tetrasaccharide trichloroacetimidate 29.
Coupling of 29 with lauryl (1!6)-linked disaccharide 26 with 3-OH gave b-(1!3)-linked hexasaccharide 30 as the major product.
Bioassay showed that in combination with the chemotherapeutic agent cyclophospamide (CPA), the hexaose 18 at a dose of 0.5–
1 mg/kg substantially increased the inhibition of S180 for CPA, but decreased the toxicity caused by CPA. Some of these oligo-
saccharides also inhibited U14 noumenal tumor in mice effectively.
# 2003 Elsevier Science Ltd. All rights reserved.
Introduction
that only higher molecular-weight fractions (MW
>16,000) obtained from partial hydrolysis of lentinan
with formic acid showed antitumor activity.4 We hypo-
thesize that the activity of lentinan is dependent upon its
basic structure-oligosaccharide unit rather than its
macroscopical morphology. It was reported that the b-
(1!3)-branched b-(1!6) polysaccharides (MW ꢀ106)
of mycelial walls of the fungus Phytophthora mega-
sperma f. sp. Glycinea can induce the formation of phy-
toalexins in soybean, and the heptasaccharide b-d-Glcp-
(1!6)-[b-d-Glcp-(1!3)-]b-d-Glcp-(1!6)-b-d-Glcp-(1!6)
-[b-d-Glcp-(1!3)-]b-d-Glcp-(1!6)-d-Glcp and the hex-
b-(1!6)-Branched b-(1!3) gluco-oligosaccharides are
the common structure characteristic of many biologi-
cally active polysaccharides from traditionally herbal
medicines such as Ganoderma lucidum, Schizophyllum
commune and Lentinus edodes.1 Of them, poly-
saccharides from Lentinus edodes called lentinan have
the most strong antitumor effect, particularly, for Sar-
coma-180in mice, and have been used as an antitumor
and immune reinforce medicine in Japan and China for
many years. Clinically, lentinan has proved effective
with chemotherapeutic agents for patients with recur-
rent gastric and colorectal cancer.2 Some physicochem-
ical and immunopharmacological investigations showed
that the antitumor activity of these glucans may be clo-
sely related to the triplet-helix structures of the b-(1!3)-
linked backbone chains3a,b and some biological aspects
of b-glucans were reported.3c,d,e It was also reported
asaccharide
b-d-Glcp-(1!6)-[b-d-Glcp-(1!3)-]b-d-
Glcp-(1!6)-b-d-Glcp-(1!6)-[b-d-Glcp-(1!3)-]d-Glcp
present in the polysaccharides have even more strong
activity than the polysaccharides themselves.5 There-
fore, we deduce that the fragment of lentinan b-d-Glcp-
(1!3)-[b-d-Glcp-(1!6)-]b-d-Glcp-(1!3)-b-d-Glcp-(1!3)
-[b-d-Glcp-(1!6)-]b-d-Glcp-(1!3)-d-Glcp or b-d-Glcp-
(1!3)-[b-d-Glcp-(1!6)-]b-d-Glcp-(1!3)-b-d-Glcp-
(1!3)-[b-d-Glcp-(1!6)-]d-Glcp may have similar or
better biological activities compared to lentinan itself.
*Corresponding author. Tel.: +86-10-62849157; fax: +86-10-
62923563; e-mail: fzkong@mail.rcees.ac.cn
0968-0896/03/$ - see front matter # 2003 Elsevier Science Ltd. All rights reserved.
doi:10.1016/S0968-0896(03)00118-4

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J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
Here we would like to disclose the synthesis and biolo-
gical activity of lentinan hexasaccharide and its analo-
gues with an a-linkage in the backbone.
readily obtained by diacetonation of d-glucose accord-
ing the standard method. Compound 4 was prepared
from allylation of 3 followed by selective removal of
5,6-O-isopropylidation with 90% HOAc. Compound 5
was obtained as white crystals from the allylation of 3
followed by deisopropylidenation, acetylation, 1-O-
deacetylation and trichloroacetimidation. The coupling
of 1,2:5,6-di-O-isopropylidene-a-d-glucofuranose 3 with
perbenzoyl glucosyl trichloroacetimidate 2 in the pre-
sence of trimethylsilyl trifluoromethanesulfonate
(TMSOTf) as the catalyst, followed by selective 5,6-O-
deacetonation afforded b-(1!3)-linked disaccharide 7
as crystals in high yield (81% over the two steps,
Scheme 2).
Results and Discussion
Preparation of oligosaccharides is more difficult and
complex compared to the synthesis of other biopoly-
mers such as peptides and nucleic acids. Most glycosy-
lation methods are extremely sensitive to structural
variations in the glycosyl donor-acceptor pairs.6 Reac-
tion conditions that provide excellent yields with one
donor–acceptor pair may give virtually no product for
another donor–acceptor pair. Furthermore, the stereo-
chemical outcome is often difficult to predict. Our
synthesis of the b-(1!6)-branched b-(1!3) glucohex-
aose again showed complexity of the oligosaccharide
synthesis as described below.
Condensation of 7 with 2 catalyzed by TMSOTf, regio-
and stereoselectively gave one of the key intermediates:
the 3,6-branched trisaccharides 8 in excellent yield
(90%). Removal of the 1,2-O-isopropylidene of 8 in
80% HOAc followed by acetylation with acetic anhy-
dride in pyridine, selective 1-O-deacetylation with
ammonia in THF–CH3OH, and subsequent treatment
with trichloroacetonitrile in the presence of K2CO3
afforded the trisaccharide glycosyl donor 9 in good yield
(71% over the four steps). Using the same procedure as
described for the preparation of 8, another key tri-
saccharide 12 was obtained from compounds 5, 3, and
2. Acetylation of 12 with acetic anhydride in pyridine
followed by deallylation with PdCl2 in CH3OH–CH2Cl2
afforded the trisaccharide glycosyl acceptor 13 in high
yield (85%). Under the same conditions as described for
the synthesis of 9 from 8, compound 14 was obtained in
good yield (71% over the four steps) from 12. Coupling
of 14 with C12H25OH followed by deallylation afforded
another trisaccharide glycosyl acceptor 16 in good yield
(67% over the two steps). Coupling of the trisaccharide
glycosyl donor 9 with either trisaccharide glycosyl
acceptor 13 or 16, catalyzed by TMSOTf in CH2Cl2, did
not afford the expected b-linked haxasaccharides, but
gave the pure a-linked haxasaccharides 17 and 19,
respectively, in excellent yields (86% for 17, 84% for
19). In the synthesis, the coupling of 9 with 13 gave an
orthoester intermediate 170 that was isolated and well
identified, and could be transformed to 17 in the pre-
sence of catalytic TMSOTf. Deisopropylidenation of 17
in 80% HOAc, followed by deacylation in an ammonia-
saturated solution in 1:1 CH2Cl2–CH3OH, furnished the
free hexasaccharide 18 as an amorphous white solid in
92% yield (over the two steps), and deacylation of 19
gave 20 as an amorphous white solid in 95% yield.
More than 3 years ago we invented an efficient method
for the synthesis of 3,6-branched b-linked gluco-oligo-
saccharids using 1,2:5,6-di-O-isopropylidene-a-d-gluco-
furanose as the starting glycosyl acceptor.7 The
synthesis of b-(1!3)-branched b-(1!6)-linked gluco-
hexaose phytoalexin elicitor on a 100 g scale was
achieved in our laboratory and higher oligosaccharides
of the elicitor including the hepta-, nona-, dodeca- and
tetradecasaccharids were also readily synthesized by the
developed strategy.8 However, when the same strategy
was used to prepare the b-(1!6)-branched b-(1!3)-
linked glucohexaoses from a trisaccharide donor and a
trisaccharide acceptor, the desired b-linked target was
not obtained, but a-(1!3)-linked analogue was the
product.9,10
In our synthesis, 2,3,4,6-tetra-O-benzoyl-a-d-glucopyr-
anosyl trichloroacetimidate 2, 1,2:5,6-di-O-isopropylidene-
a-d-glucofuranose 3, 3-O-allyl-1,2-O-isopropylidene-a-d-
glucofuranose 4 and 2,4,6-tri-O-acetyl-3-O-allyl-a-d-glu-
copyranosyl trichloroacetimidate 5 were the starting
materials and the trisaccharides 8, 12 and the disaccharide
21 were the key intermediates. Compound 2 was prepared
as fine crystals via benzoylation of d-glucose followed by
1-O-debenzoylation with ammonia in THF–CH3OH and
trichloroacetimidation (Scheme 1). Compound 3 was
During our research, we found that the stereo-selectivity
for the formation of (1!3) linked glucosyl bonds from
the donor with C2 ester capable of neighboring group
participation is influenced by the structure of both the
glycosyl acceptors and glycosyl donors and some of
these research results have been published recently.11
For the synthesis of fully b-linked oligosaccharides,
4,6-O-benzylidene-b-d-glucopyranose was used as the
acceptor since its derivatives often give the b-(1!3)
linked glucosyl bonds exclusively as reported.12 Thus
condensation of 2 with 4, catalyzed by TMSOTf, regio-
selectively gave the key disaccharide 21 in excellent yield
Scheme 1. Keys: (a) (i) PhCOCl, pyridine and toluene, 70 ꢁC, 8 h; (ii)
3:1 (v/v) THF–CH3OH, 1.5 N NH3, rt, 12 h; (iii) CH2Cl2, CCl3CN and
K2CO3, rt, 24 h. (b) (i) AllBr, NaH and DMF, rt, 2 h; (ii) 90% HOAc,
40 ꢁC, 24 h. (c) (i) AllBr, NaH and DMF, rt, 2 h; (ii) 80% HOAc,
reflux, 4 h; (iii) Ac2O–Pyridine, rt, 2 h; (iv) 3:1 (v/v) THF–CH3OH,
1.5 N NH3, rt, 3 h; (v) CH2Cl2, CCl3CN and K2CO3, rt, 24 h.

J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
2195
Scheme 2. Keys: (a) TMSOTf, 4 A MS. CH2Cl2, rt, 3 h. (b) 90% HOAc, 40 ꢁC, 24 h. (c) (i) 80% HOAc, reflux, 5 h; (ii) Ac2O–Pyridine, rt, 2 h; (iii)
THF–CH3OH, 1.5 N NH3, rt, 3 h; (iv) CH2Cl2, CCl3CN and K2CO3, rt, 24 h. (d) (i) Ac2O–Pyridine, rt, 2 h; (ii) PdCl2, CH2Cl2–CH3OH, rt, 5 h. (e)
PdCl2 in CH2Cl2–CH3OH, rt, 5 h. (f) (i) 80% HOAc, reflux, 6 h; (ii) CH2Cl2–CH3OH saturated with ammonia, rt, 24 h. (g) TMSOTf, 4 A MS,
CH2Cl2, rt, 20min. (h) TMSOTf, 4 A MS, CH2Cl2, rt, 3 h. (m) CH2Cl2–CH3OH saturated with ammonia, rt, 24 h.

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J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
(84%) (Scheme 3). The disaccharide glycosyl acceptor
23 was subsequently obtained by 5-O-benzoylation of 21
followed by 3-O-deallylation in 78% yield. Lauryl 2,3,4,6
-tetra-O-benzoyl-b-d-glucopyranosyl-(1!6)-2,4-di-O-
acetyl-b-d-glucopyranoside (26) as another disaccharide
glycosyl acceptor was prepared from condensation of 24
with lauryl alcohol followed by deallylation. Regio- and
stereoselective coupling of 9 with 4-methoxyphenyl
4,6-O-benzylidene-b-d-glucopyranoside (27)13 afforded
the desired tetrasaccharide 28 (77% yield) which was
then converted to the tetrasaccharide glycosyl donor 29
by removal of the 4,6-O-benzylidene of 28 in 90%
HOAc followed by acetylation, demethoxyphenylation
with (NH4)2Ce(NO3)6 in CH3CN-H2O,14 and sub-
sequent trichloroacetimidation in 71% yield (over the
four steps). However, coupling of 29 with 23, catalyzed
by TMSOTf in CH2Cl2, gave nothing except the
decomposed product of glycosyl donor 29 and the
unchanged glycosyl acceptor 23. Condensation of 29
with 26 afforded a mixture of the b-(1!6)-branched
b-(1!3)-linked glucohexatose 30 (38% yield) and its iso-
mer containing an a-(1!3) linked bond 31 (16% yield).
At the initial stage of the coupling, an orthoester inter-
mediate 300 was obtained and could be isolated. With
addition of the extra catalyst TMSOTf or elongation of
the reaction time, 300 was transformed to a mixture of 30
Scheme 3. Keys: (a) TMSOTf, 4 A MS, CH2Cl2, rt, 3 h. (b) PhCOCl, pyridine, rt, 2 h. (c) PdCl2 in CH2Cl2–CH3OH, rt, 3 h. (d) (i) 80% HOAc,
reflux, 5 h; (ii) Ac2O–Pyridine, rt, 2 h; (iii) THF–CH3OH, 1.5 N NH3, rt, 3 h; (iv) CH2Cl2, CCl3CN and K2CO3, rt, 24 h. (e) (i) 90% HOAc, 40 ꢁC,
24 h; (ii) Ac2O–Pyridine, rt, 2 h; (iii) (NH4)2Ce(NO3)6 in CH3CN–H2O, rt, 0.5 h; (iv) CH2Cl2, CCl3CN and K2CO3, rt, 24 h. (f) CH2Cl2–CH3OH
saturated with ammonia, rt, 24 h.

J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
2197
and 31. It seemed that transformation of the orthoester
300 to the corresponding oligosaccharides 31 and 32
went through two different paths as indicated in Scheme
4. Path 1 (!30) was the normal rearrangement leading
to trans glycosylation, while path 2 (!31) was the unu-
sual one caused by C1-O1 breaking of the orthoester.11
Finally, compounds 32 and 33 were obtained by depro-
tection of 30 and 31, respectively.
Table 2. Inhibition of U14 tumor by 18, 20, allyl heptaoside, and
CPA
Groups
Weight (g) Quotient of noumenal Inhibition
tumor (%)
(%)
U14 cell line group
Group 1: treatment 27.40Æ1.902.63
29.33Æ1.65
4.21Æ1.91
Æ0.90
29.41
58.43
58.43
with 18 (10mg/kg)
Group 2: treatment 27.45Æ2.67
with 20 (5 mg/kg)
1.54Æ0.56**
1.51Æ0.41**
Group 3: treatment 28.10Æ1.79
Bioassay showed that in combination with the chemo-
therapeutic agent cyclophospamide (CPA), the hexaose
18 at a dose of 0.5–1 mg/kg subatantially increased the
inhibition of CPA to S180, but decreased the toxicity
caused by CPA as indicated from the amount of nucle-
ated cell in bone marrow (Table 1).
with allyl heptaoside
(5 mg/kg)
Group 4: treatment 28.00Æ1.78
1.86Æ0.73*
39.22
with CPA (70mg/kg)
Compared to U14 cell line group, *p<0.05 **p<0.01.
Experimental
Inhibition of U14 noumenal tumor by the synthetic
olgosaccharides was also investigated indicating that the
allyl heptaoside, a fully b-linked lentinan repeating
unit,13 and lauryl hexaoside 20 were potential ther-
apeutic agents for cancer treatment (Table 2).
Melting points were determined with a ‘Mel-Temp’
apparatus. Optical rotations were determined at 25 ꢁC
with digital polarimeter. The NMR spectra were recor-
ded in CDCl3 with TMS internal standard or D2O with
ethanol as standard on ARX 400 MHz. Mass spectra
were recorded on an autospec mass spectrometer using
ESI technique to introduce the sample. Elemental ana-
lyses were done on elemental analyzer model 1108 EA.
Thin-layer chromatography (TLC) was performed on
silica gel HF254 with detection by charring with 30% (v/
Now in our laboratory, a lot of derivatives of the
(1!6)-branched (1!3)-linked glucohexaoses are in
preparation, and interesting results about the structure–
activity relationship of the newly discovered biologically
active oligosaccharides including 32 and 33 will be
reported in due course.
Scheme 4.
Table 1. Inhibition of S180 and effect on nucleated cell in bone marrow for the hexaose 18 and 18+CPA
Groups
Weight (g)
Amount of nucleated cell (Â 107)
Amount of tumor cell (Â 107)
Control group
S180 Cell line group
22.29Æ0.95
22.00Æ1.00
21.86Æ0.90
22.00 Æ1.00
21.71Æ0.95
22.00Æ0.82
21.86 Æ0.69
7.45Æ2.35###
7.65Æ3.05
2.94Æ0.54
Group 1: treatment with 18 (1 mg/kg)
Group 2: treatment with 18 (1 mg/kg)+CPA (50mg/kg)
Group 3: treatment with 18 (0.5 mg/kg)
Group 4: treatment with 18 (0.5 mg/kg)+CPA (50 mg/kg)
Group 5 treatment with CPA (50mg/kg)
6.70Æ1.95###^
3.60Æ1.15#
2.01Æ0.56*
1.53Æ0.85***
2.38Æ0.61*
1.57Æ0.62**
2.01Æ0.53*
6.80Æ2.00###^
3.55Æ1.55##
1.50Æ0.45##
Compared to group 5, #p<0.05. ##p<0.01. ###p<0.001. Comparison of a kind of drug to the combined two, ^p<0.05. *Compared to the S180 cell
line group, p<0.05. **p<0.01. ***p<0.001.

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J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
v) H2SO4 in MeOH or in some cases by a UV detector.
Column chromatography was conducted by elution of a
column (10Â240mm, 18 Â300 mm, 35Â400 mm) of silica
gel (100–200 mesh) with EtOAc–petroleum ether
(60–90 ꢁC) as the eluent. Solutions were concentrated at
<60 ꢁC under diminished pressure. Dry solvents were
distilled over CaH2 and stored over molecular sieves.
petroleum ether–EtOAc) indicated that the reaction was
complete. The mixture was filtrated, the solution was
concentrated under reduced pressure, and the residue
was decolorized by passing through a short silica-gel
column with 3:1 petroleum ether–EtOAc as the eluent
and 2,3,4,6-tetra-O-benzyol-a-d-glucopyranosyl tri-
chloroacetimidate (2) (46 g, 56% for three steps) was
crystallized from 3:1 petroleum ether–EtOAc as white
crystals.15
Bioassay for inhibition of S180
Cyclophosphamide was purchased from Shanghai Hua-
lian Pharmaceutical Co. Ltd. (permission No. Hu Med-
icines 95-012034). Animals: 50 Kunming male mice
(weight 20Æ2 g, grade II) were obtained from the
Experimental Animal Center, the Chinese Academy of
Medicines. S180 cell line was provided by the Depart-
ment of Tumorology, Institute of Pharmacy, the Chi-
nese Academy of Medicines. The mice were randomly
divided into seven groups according to weight, and into
each of them was planted S180 cell line of ascites cancer
0.2 mL (amount of cell 7.9 Â 106) in their euterocelia by
intraperitoneal except for the control group mice. At
24 h after the planting, the mice were treated with the
synthetic oligosaccharides or cyclophosphamide. The
oligosaccharide was dissolved in saline at 0.1 mg/mL,
given with a dose of 0.5 or 1 mg/kg/day to the mice
within three consecutive days. Cyclophosphamide was
dissolved in saline at 4 mg/mL, and given to the mice at
a dose of 20mg/kg by intraperitoneal at 24 h after the
planting, and 30mg/kg at 48 h after the planting. For
inhibition of U14, the same mice were used and divided
into seven groups. Into each of the mice was planted
U14 cell line 0.2 mL (amount of cell 4 Â 106) by intra-
peritoneal. At 24 h after the planting, the mice were
treated with the synthetic oligosaccharides or cyclophos-
phamide. The oligosaccharides 18 and 20 were dissolved
in saline at 1 mg/mL, given with a dose of 10mg/kg/d to
the mice within ten consecutive days (0.2 mL/day, ip).
Allyl heptaoside was dissolved in saline at 1 mg/mL,
given with a dose of 5 mg/kg/day to the mice within ten
consecutive days (0.1 mL/day ip). Cyclophosphamide
was dissolved in saline at 6 mg/mL, and given to the
mice at a dose of 30mg/kg by intraperitoneal at 24 h
after the planting, and 20mg/kg at 72 h and 120h after
the planting, respectively.
3-O-Allyl-1,2-O-isopropylidene-ꢁ-D-glucopyranose (4).
To a solution of 3 (10.0 g, 38.4 mmol) in dry DMF
(50mL), AllBr (3.7 mL, 42.2 mmol) and NaH (3.0g,
49% in oil, 61.4 mmol) were added under cooling with
an ice bath. The mixture was stirred for 2 h 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 resulting resi-
due was directly dissolved in 90% aceꢁtic acid solution
(80mL). The mixture was kept at 40 C for 24 h and
then concentrated to a residue under reduced pressure.
The residue was purified by flash chromatography (2:1
petroleum ether–EtOAc) to give 4 (7.1 g, 71% for two
steps) as syrup. [a]D +14ꢁ (c 1.2, CHCl3);
1HNMR(400 MHz, CDCl3) d 5.92 (d, 1H, J=3.8 Hz),
5.90(m, 1H), 5.32 (dd, 1H, J=1.3, 17.1 Hz), 5.23 (dd,
1H, J=1.3 Hz, 10.4 Hz), 4.58 (d, 1H, J=3.8 Hz),
4.22–4.03 (m, 5H), 3.84 (dd, 1H, J=3.4, 11.4 Hz), 3.74
(dd, 1H, J=5.5, 11.4 Hz). Anal. calcd for C12H20O6: C,
55.37; H, 7.74. Found: C, 55.81; H, 7.80.
3-O-Allyl-2,4,6-tri-O-acetyl-ꢁ-D-glucopyranosyl trichloro-
acetimidate (5). To a solution of 3 (50.0 g, 192 mmol) in
dry DMF (160mL), AllBr (18.6 mL, 211 mmol) and
NaH (15.0g, 49% in oil, 307 mmol) were added under
cooling with an ice bath. The mixture was stirred for 2 h
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
resulting residue was directly dissolved in 80% aqueous
acetic acid solution (300 mL), and the mixture was
heated under reflux for 4 h. The mixture was con-
centrated, and the residue was treated with acetic anhy-
dride (250mL) in pyridine (280mL) for 2 h at rt. The
acetylated sugar was dissolved in a 1.5 N solution of
NH3 in 3:1 THF–CH3OH (300 mL), and the solution
was kept at rt for 3 h, at the end of which time TLC (3:1
petroleum ether–EtOAc) indicated that the reaction was
complete. The mixture was concentrated under reduced
pressure, and the residue was dissolved in a solution of
CH2Cl2 (200 mL) and CCl3CN (39 mL, 384 mmol) con-
taining K2CO3 (50g, 362 mmol). The reaction mixture
was stirred for 24 h at rt. After filtering the mixture, the
filtration and washings were concentrated, and the resi-
due was subjected to column chromatography with 3:1
petroleum ether–EtOAc as the eluent to give 5 (38.4 g,
54% for four steps) as a white crystals. Mp 71–73 ꢁC;
[a]D +0.15ꢁ (c 1.5, CHCl3); 1HNMR (400 MHz,
CDCl3) d 8.67 (s, 1H), 6.52 (d, 1H, J=3.6 Hz), 5.80(m,
1H), 5.24 (dd, 1H, J=1.3, 17.1 Hz), 5.18–5.13 (m, 2H),
5.03 (dd, J=3.6, 13.6 Hz), 4.24–4.18 (m, 2H), 4.13–4.08
2,3,4,6-Tetra-O-benzoyl-ꢁ-D-glucopyranosyl trichloroa-
cetimidate (2). BzCl (68 mL, 583 mmol) was added to a
solution of d-glucose (1) (20g, 111 mmol) in toluene
(250mL) and pyridine (47.3 mL, 585 mmol) over 1 h and
the temperature was kept at 70 ꢁC, and then the mixture
was stirred at 70 ꢁC for further 7 h. Filtration of pyr-
idinum hydrochloride salt and concentration of the fil-
trate gave a residue which was directly dissolved in a
1.5 N solution of NH3 in THF (350mL) and CH 3OH
(210mL). The solution was kept at room temperature
for 12 h, at the end of which time TLC (3:1 petroleum
ether–EtOAc) indicated that the reaction was complete.
The mixture was concentrated under reduced pressure,
and the residue was dissolved in a solution of CH2Cl2
(100 mL) and CCl3CN (12 mL, 120mmol) containing
K2CO3 (30g, 217 mmol ). The reaction mixture was
stirred for 24 h at rt, at the end of which time TLC (3:1

J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
2199
(m, 3H), 3.97 (t, 1H, J=9.6 Hz), 2.10, 2.08, 2.07 (3 s,
9H). Anal. calcd for C17H22NO9Cl3: C, 55.13; H, 5.99.
Found: C, 55.62; H, 5.90.
chromatography to give 9 (8.0g, 71% for four steps).
[a]D +23.3ꢁ (c 1.0, CHCl3); 1H NMR (400 MHz,
CDCl3) d 8.33 (s, 1H), 8.07–7.19 (m, 40H), 6.21 (d, 1H,
J=3.6), 5.91 (t, 1H, J=9.6 Hz), 5.85 (t, 1H, J=9.6 Hz),
5.62 (t, 1H, J=9.6 Hz), 5.61 (t, 1H, J=9.6 Hz), 5.46 (dd,
1H, J=7.9, 9.6 Hz), 5.42 (dd, 1H, J=7.9, 9.6 Hz), 4.97
(d, 1H, J=7.9 Hz), 4.96 (d, 1H, J=7.9 Hz), 4.85 (t, 1H,
J=9.5 Hz), 4.67–4.59 (m, 3H), 4.50–4.37 (m, 2H),
4.19–4.02 (m, 4H), 3.91(dd, 1H), 3.69 (dd, 1H), 1.94,
1.78 (2 s, 6H). Anal. calcd for C80H68NO26Cl3: C, 61.37;
H, 4.38. Found: C, 61.93; H, 4.51.
2,3,4,6-Tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!3)-1,2-
O-isopropylidene-ꢁ-D-glucofuranose (7). To a stirred
solution of 3 (10g, 38 mmol) and 2 (26 g, 35 mmol) in
CH2Cl2 (100 mL) was added TMSOTf (30 mL) at room
temperature. After 3 h, triethylamine was added to the
solution to quench the reaction. The solution was con-
centrated, the resulting residue was directly dissolved in
90% aqueous acetic acid solution (200 mL). The mix-
ture was kept at 40 ꢁC for 24 h and then concentrated to
a residue under reduced pressure. The resulting residue
was subjected to a short silica-gel column to give com-
pound 7 (22.6 g, 81% for two steps). Mp 120–123 ꢁC;
3-O-Allyl-2,4,6-tri-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-
1,2-O-isopropylidene-ꢁ-D-glucofuranose (11). To a stir-
red solution of 3 (10g, 38.5 mmol) and 5 (14.1 g,
38.0mmol) in CH 2Cl2 (100 mL) was added TMSOTf
(35 mL) at room temperature. After 3 h, triethylamine
was added to the solution to quench the reaction. The
solution was concentrated, the resulting residue was
directly dissolved in 90% aqueous acetic acid solution
[a]D +14ꢁ (c 2.5, CHCl3). HNMR (400 MHz, CDCl3)
1
d 8.11–7.28 (m, 20H), 5.94 (t, 1H, J=9.7 Hz), 5.72 (t,
1H, J=9.7 Hz), 5.54 (dd, 1H, J=7.9, 9.7 Hz), 5.53 (d,
1H, J=3.6 Hz), 5.03 (d, 1H, J=7.9 Hz), 4.84 (dd, 1H,
J=3.6, 11.9 Hz), 4.42 (dd, 1H, J=4.3, 11.9 Hz), 4.41(d,
1H, J=2.6 Hz), 4.24–4.23 (m, 2H), 4.16 (dd, 1H, J=2.6,
8.8 Hz), 4.02 (m, 1H), 3.83 (dd, 1H, J=3.2, 11.4 Hz),
3.67 (dd, 1H, J=6.0, 11.4 Hz), 1.44, 1.09 (2 s, 6H).
Anal. calcd for C43H42O15: C, 64.66; H, 5.30. Found: C,
64.94; H, 5.25.
ꢁ
(150mL). The mixture was kept at 40 C for 24 h and
then concentrated to a residue under reduced pressure.
The resulting residue was subjected to a short silica-gel
column to give compound 11 (15 g, 72% for two steps)
as a white crystals. Mp 91–93 ꢁC; [a]D +42 (c 1.8,
1
CHCl3). H NMR (400 MHz, CDCl3): d 5.77 (d, 1H,
J=3.6 Hz), 5.71 (m, 1H), 5.14 (dd, 1H), 5.08 (dd, 1H),
4.96 (t, 1H, J=9.6 Hz), 4.90(dd, 1H, J=7.9, 9.4 Hz),
4.53 (d, 1H, J=7.9 Hz), 4.30(d, 1H), 4.22 (d, 1H), 4.12–
4.00 (m, 5H), 3.87 (m, 1H), 3.77 (dd, 1H), 3.60–3.58 (m,
2H), 3.54 (t, 1H, J=9.4 Hz), 2.05, 2.03, 2.02 (3 s, 9H),
1.42, 1.25 (2 s, 6H). Anal. calcd for C24H36O14: C, 52.55;
H, 6.61. Found: C, 52.81; H, 6.52.
2,3,4,6-Tetra-O-benzoyl-ꢀ-D -glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-1,2-
O-isopropylidene-ꢁ-D-glucofuranose (8). To a stirred
solution of 7 (8 g, 10mmol) and 2 (8 g, 10.8 mmol) in
CH2Cl2 (60mL) was added TMSOTf (30 mL) at room
temperature. After 3 h, triethylamine was added to the
solution to quench the reaction. The solution was con-
centrated, the residue was subjected to column chroma-
tography with 1.5:1 petroleum ether–ethyl acetate as the
eluent to give 8 (12.4 g, 90%). [a]D +19.3ꢁ (c 1.0,
3-O-Allyl-2,4,6-tri-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-1,2-
O-isopropylidene-ꢁ-D-glucofuranose (12). To a stirred
solution of 11 (20g, 36.5 mmol) and 2 (27.4 g, 37 mmol)
in CH2Cl2 (150mL) was added TMSOTf (40 mL) at
room temperature. After 3 h, triethylamine was added
to the solution to quench the reaction. The solution was
concentrated, the residue was subjected to column
chromatography with 1.5:1 petroleum ether–ethyl ace-
tate as the eluent to give the trisaccharide 12 (37.0g,
1
CHCl3); H NMR (400 MHz, CDCl3): d 8.06–7.28 (m,
40H), 5.88 (t, 1H, J=9.7 Hz), 5.87 (t, 1H, J=9.7 Hz),
5.69 (t, 1H, J=9.7 Hz), 5.64 (t, 1H, J=9.7 Hz), 5.53 (dd,
1H, J=7.9, 9.7 Hz), 5.43 (dd, 1H, J=7.9, 9.7 Hz), 5.41
(d, 1H, J=3.5 Hz), 4.96 (d, 1H, J=7.9 Hz), 4.93 (d, 1H,
J=7.9 Hz), 4.71–4.67 (m, 2H), 4.48 (dd, 1H, J=4.9,
12.2 Hz), 4.35 (dd, 1H, J=4.9, 12.2 Hz), 4.34–3.65 (m,
8H), 1.26, 1.03 (2 s, 6H). Anal. calcd for C77H68O24: C,
67.15; H, 4.98. Found: C, 66.89; H, 5.09.
1
90%). [a]D +16.1 (c 1.7, CHCl3); H NMR (400 MHz,
CDCl3): d 8.02–7.27 (m, 20H), 5.89 (t, 1H, J=9.6 Hz),
5.76 (d, 1H, J=3.7 Hz), 5.75 (m, 1H), 5.69 (t, 1H,
J=9.6 Hz), 5.55 (dd, 1H, J=7.9, 9.6 Hz), 5.20(dd, 1H),
5.14 (dd, 1H), 5.01 (d, 1H, J=7.9 Hz), 5.00 (t, 1H,
J=9.7 Hz), 4.89 (dd, 1H, J=7.9, 9.3 Hz), 4.67 (dd, 1H),
4.52 (d, 1H, J=7.9 Hz), 4.48 (dd, 1H), 4.33 (d, 1H), 4.23
(d, 1H), 4.17–4.01 (m, 8H), 3.80 (m, 1H), 3.60–3.54 (m,
2H), 2.08, 2.07, 2.02 (3 s, 9H), 1.32, 1.26 (2 s, 6H). Anal.
calcd for C58H62O23: C, 61.81; H, 5.54. Found: C, 61.46;
H, 5.61.
2,3,4,6-Tetra-O-benzoyl-ꢀ-D -glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-ꢁ-D-glucopyranosyl trichloroacetimidate (9).
Compound 8 (10g, 7.2 mmol) was added to 80% aqu-
eous acetic acid solution (100 mL) and the mixture was
heated under reflux for 5 h. The mixture was con-
centrated, and the residue was acetylated with acetic
anhydride (20mL) in pyridine (20mL) for 2 h at rt. The
resultant trisaccharide was dissolved in a 1.5 N solution of
NH3 in 3:1 THF–CH3OH (100mL), and the solution was
kept at rt for 3 h. The solution was concentrated, the resi-
due was dissolved in CH2Cl2 (50mL). To the solution
were added K2CO3 (2 g, 14.5 mmol) and CCl3CN (1.4 mL,
14.4 mmol), and the mixture was stirred at rt for 24h.
Filtering the mixture, the filtration and washings were
concentrated, and the residue was subjected to column
2,4,6-Tri-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-[2,3,4,6-
tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-5-O-acetyl-
1,2-O-isopropylidene-ꢁ-D-glucofuranose (13). The com-
pound 12 (25 g, 22.2 mmol) was acetylated with acetic
anhydride (20mL) in pyridine (20mL) at rt for 2 h. The
mixture was diluted with dichloromethane, washed with M
HCl, water, and satd aq sodium bicarbonate subsequently.

2200
J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
The organic layer was combined, dried, and con-
centrated. The residue was dissolved in a solution of
CH3OH (250mL) containing PdCl 2 (100 mg). The mix-
ture was stirred for 5 h at rt, at the end of which time
TLC (2:1 petroleum ether–EtOAc) indicated that the
reaction was complete. The mixture was filtered, and the
filtrate was concentrated, and the residue was passed a
silica-gel column with 2:1 petroleum ether–EtOAc as the
eluent to give 13 (21.3 g, 85%). [a]D +26.6 (c 2.1,
reduced pressure to dryness. Purification by column
chromatography (3:1 petroleum ether–EtOAc) gave 15
(3.66 g, 80%) as a syrup: [a]D +32.1ꢁ (c 1.0, CHCl3); 1H
NMR (400 MHz, CDCl3): d 8.03–7.27 (m, 20H), 5.88 (t,
1H, J=9.8 Hz), 5.71 (m, 1H), 5.67 (t, 1H, J=9.2 Hz),
5.50(dd, 1H, J=8.0, 9.6 Hz), 5.16 (dd, 1H), 5.11 (dd,
1H), 4,99 (t, 1H, J=9.2 Hz), 4.94 (d, 1H, J=8.0Hz),
4.85 (d, 1H, J=8.0Hz), 4.84 (d, 1H, J=8.0Hz), 4.64
(m, 1H), 4.46 (m, 2H), 4.25 (dd, 1H, J=12.4, 3.2 Hz),
4.15–3.43 (m, 11H), 3.01 (m, 1H), 2.07, 2.06, 2.05, 2.04,
1.95 (5 s, 15H), 1.30–1.18 (m, 20H), 0.88 (t, 3H). Anal.
calcd for C71H86O25: C, 63.67; H, 6.47. Found: C, 64.01;
H, 6.20.
1
CHCl3); H NMR (400 MHz, CDCl3): d 8.07–7.27 (m,
20H), 5.87 (t, 1H, J=9.5 Hz), 5.74 (t, 1H, J=9.7 Hz),
5.73 (d, 1H, J=3.5 Hz), 5.54 (dd, 1H, J=7.9, 9.5 Hz),
5.16 (m, 1H), 4.99 (t, 1H, J=9.6 Hz), 4.87 (d, 1H,
J=7.9 Hz), 4.81 (dd, 1H, J=7.8, 9.3 Hz), 4.50(d, 1H,
J=7.8 Hz), 4.43–4.39 (m, 2H), 4.30–4.27 (m, 2H),
4.22–4.06 (m, 5H), 3.81 (m, 1H), 3.67 (t, 1H, J=9.6 Hz),
3.57 (m, 1H), 2.09, 2.08, 2.03, 1.73 (4 s, 12H), 1.37, 1.28
(2 s, 6H). Anal. calcd for C57H60O24: C, 60.64; H, 5.36.
Found: C, 60.98; H, 5.28.
Lauryl
2,4,6-tri-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-ꢀ-D-glucopyranoside (16). Compound 15
(3.4 g, 2.54 mmol) was dissolved in a solution of
CH3OH (70mL) and CH 2Cl2 (10mL). To the solution
was added PdCl2 (25 mg), and the mixture was stirred
for 5 h at rt, at the end of which time TLC (2:1 petro-
leum ether–EtOAc) indicated that the reaction was
complete. The mixture was filtered, and the filtrate was
concentrated, and the residue was passed through a
silica-gel column with 2:1 petroleum ether–EtOAc as the
eluent to give 16 (2.77 g, 84%). [a]D +19.3 (c 2.0,
3-O-Allyl-2,4,6-tri-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-ꢁ-D-glucopyranosyl
trichloroacetimidate
(14). Compound 12 (14 g, 12.4 mmol) was added to
80% acetic acid solution (150 mL) and the mixture was
heated under reflux for 5 h. The mixture was con-
centrated, and the residue was acetylated with acetic
anhydride (20mL) in pyridine (20mL) for 2 h at rt. The
mixture was diluted with dichloromethane, washed with
M HCl, water, and satd aq sodium bicarbonate subse-
quently. The organic layer was combined, dried, and
concentrated. The trisaccharide residue was dissolved in
a 1.5 N solution of NH3 in 3:1 THF–CH3OH (150mL),
and the solution was kept at rt. After 3 h, the solution
was concentrated, and the residue was dissolved in
CH2Cl2 (80mL). To the solution were added K 2CO3
(5 g, 36.2 mmol) and CCl3CN (2.0mL, 20mmol), and
the mixture was stirred at rt for 24 h. Filtering the mix-
ture, the filtration and washings were concentrated, and
the residue was subjected to column chromatography to
give the trisaccharide donor 14 (11.6 g, 71% for four
1
CHCl3); H NMR (400 MHz, CDCl3) d 8.03–7.27 (m,
20H), 5.88 (t, 1H, J=9.7 Hz), 5.67 (t, 1H, J=9.6 Hz),
5.50(dd, 1H, J=7.9, 9.6 Hz), 4.93 (d, 1H, J=7.9 Hz),
4.89 (t, 1H, J=9.6 Hz), 4.87 (t, 1H, J=9.7 Hz),
4.70–4.62 (m, 2H), 4.47 (dd, 1H), 4.45 (d, 1H,
J=7.9 Hz), 4.32 (dd, 1H), 4.16–4.14 (m, 2H), 4.03 (dd,
1H), 3.94 (d, 1H), 3.75 (t, 1H), 3.67–3.57 (m, 3H), 3.45
(m, 1H), 3.00 (m, 1H), 2.09, 2.08, 2.07, 2.06, 1.96 (5 s,
15H), 1.35–1.07 (m, 20H), 0.89 (t, 3H). Anal. calcd for
C68H82O25: C, 62.86; H, 6.36. Found: C, 62.41; H, 6.49.
2,3,4,6-Tetra-O-benzoyl-ꢀ-D -glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-ꢁ-D-glucopyranosyl-(1!3)-2,4,6-tri-O-acetyl
-ꢀ-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-ꢀ-D
-glucopyranosyl-(1!6)]-5-O-acetyl-1,2-O-isopropylidine-
ꢁ-D-glucopyranoside (17). To a stirred solution of 9
(7.30g, 4.66 mmol) and 13 (5.1 g, 4.5 mmol) in CH2Cl2
(60mL) was added TMSOTf (30 mL) at rt. After 3 h, the
mixture was neutralized with triethylamine and con-
centrated. The residue was purified on a silica gel col-
umn with 1.5:1 petroleum ether–EtOAc as the eluent to
give 17 (9.8 g, 86%). [a]D +18.7 (c 1.3, CHCl3); 1H
NMR (400 MHz, CDCl3): d 7.89–7.27 (m, 60H), 5.88,
5.87, 5.82, 5.75 (4 t, 4H, J=9.5 Hz), 5.72 (d, 1H,
J=3.5 Hz), 5.68, 5.63 (2 t, 2H, J=9.5 Hz,), 5.59, 5.45,
5.38 (3 dd, 3H, J=7.9, 9.5 Hz,), 5.05 (m, 1H), 4.97, 4.92
(2 d, 2H, J=7.9 Hz), 4.90(d, 1H, J=3.6 Hz), 4.84, 4.53
(2 d, 2H, J=7.9 Hz), 2.00, 1.97, 1.84, 1.83, 1.82, 1.77 (6
s, 18H), 1.37, 1.35 (2 s, 6H); 13C NMR (100 MHz,
CDCl3): d 170.01, 169.81, 169.22, 168.93, 168.78,
168.31, 165.70, 165.62, 165.52, 165.40, 165.32, 165.17,
165.02, 164.75, 164.61, 164.54, 111.92, 104.74 (C-1 for a
bond, JC-H=182.4 Hz), 100.69, 100.62, 100.48, 97.76 (4
C-1 for b bonds, JC-H=161.3–163.7 Hz), 93.62 (C-1 for
a bond, JC-H=174.8 Hz), 81.60, 78.53, 74.48, 26.43,
25.93, 20.34, 20.21, 20.21, 20.21, 20.00, 20.00. Anal.
1
steps). [a]D +62.0( c 1.2, CHCl3); H NMR (400 MHz,
CDCl3) d 8.35 (s, 1H), 8.04–7.26 (m, 20H), 6.22 (d, 1H,
J=3.5 Hz), 5.86 (t, 1H, J=9.6 Hz), 5.71 (m, 1H), 5.64
(t, 1H, J=9.6 Hz), 5.47 (dd, 1H, J=7.8, 9.6 Hz), 5.18
(dd, 1H), 5.06 (dd, 1H), 5.00 (t, 1H, J=9.6 Hz), 4.94 (d,
1H, J=7.8 Hz), 4.87 (t, 1H, J=9.7 Hz), 4.86(t, 1H,
J=9.6 Hz), 4.80(dd, 1H, J=7.9, 9.6 Hz), 4.62 (dd, 1H),
4.49 (d, 1H, J=7.9 Hz), 4.48 (dd, 1H), 4.22 (dd, 1H),
4.15–4.01 (m, 6H), 3.93 (d, 1H), 3.69–3.51 (m, 3H), 2.06,
2.05, 2.04, 2.01, 1.99 (5 s, 15H). Anal. calcd for
C61H62NO25Cl3: C, 55.69; H, 4.75. Found: C, 55.19; H,
4.58.
Lauryl 3-O-allyl-2,4,6-tri-O-acetyl-ꢀ-D-glucopyranosyl-
(1!3)-[2,3,4,6-tetra-O-benzoyl-ꢀ-D -glucopyranosyl-
(1!6)]-2,4-di-O-acetyl-ꢀ-D-glucopyranoside (15). To a
solution of 14 (4.5 g, 3.42 mmol) and lauryl alcohol
(0.93 g, 5.0 mmol) in CH2Cl2 (50mL) was added
TMSOTf (30 mL) at rt. The reaction mixture was stirred
for 3 h, at the end of which time TLC indicated that the
reaction was complete. Then the mixture was neu-
tralized with triethylamine and concentrated under

J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
2201
calcd for C135H126O49: C, 64.03; H, 5.01. Found: C,
64.51; H, 4.93.
(20). Compound 19 (1.6 g, 0.59 mmol) was dissolved in
a saturated solution of ammonia in CH2Cl2 (10mL) and
CH3OH (100 mL) at rt. After 24 h, the reaction mixture
was concentrated, and the residue was washed four
times with CH2Cl2 to afford 20 as white solid (650mg,
95%).[a]D ꢂ13.6 (c 0.1, H2O); 1H NMR (400 MHz,
D2O): d 5.33 (d, 1H, J=3.2 Hz), 4.73, 4.50, 4.46, 4.41,
4.21 (5 d, 5H, J=7.5 Hz); 4.20–3.36 (m, 38H), 1.67 (m,
2H), 1.45–1.21 (m, 18H), 0.92 (t, 3H); 13C NMR
(100 MHz, D2O): d 105.22, 105.06, 105.06, 104.95,
104.66, 101.23, 86.51, 85.55, 84.31. Anal. calcd for
C48H86O31: C, 49.74; H, 7.48. Found: C, 49.02; H, 7.89.
ESMS for C48H86O31 (1159.18): 1158.17 [M-1]+.
Orthoester 170;. To a stirred solution of 9 (3.6 g,
2.3 mmol) and 13 (2.5 g, 2.2 mmol) in CH2Cl2 (80mL)
was added TMSOTf (20 mL) at rt. After 20min the
mixture was neutralized with triethylamine and con-
centrated. The residue was purified on a silica gel col-
umn with 1.5:1 petroleum ether–EtOAc as the eluent to
give the product 170 (4.2 g, 76%). [a]D +25.9 (c 1.0,
CHCl3); 13C NMR (100 MHz, CDCl3): d 170.08, 169.65,
168.73, 168.50, 168.38, 121.77, 112.09, 104.62 (C-1 for
glucofuranose a bond, JC-H=182.4 Hz), 101.12, 100.59,
100.56, 98.00 (4 C-1 for b bonds, JC-H=161.3–
163.7 Hz), 96.32 (C-1 for a bond, JC-H=174.8 Hz),
81.89, 78.75, 20.41, 20.22, 20.19, 20.11, 19.98. Anal.
calcd for C135H126O49: C, 64.03; H, 5.01. Found: C,
63.57; H, 4.87.
2,3,4,6-Tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)-3-O-
allyl-1,2-O-isopropylidene-ꢁ-D-glucofuranose (21). To a
stirred solution of 2 (3.02 g, 4.1 mmol) and 4 (1.04 g,
4.0mmol) in CH 2Cl2 (50mL) was added TMSOTf
(20 mL) at room temperature. After 3 h, triethylamine
was added to the solution to quench the reaction. The
solution was concentrated, the residue was subjected to
column chromatography with 3:1 petroleum ether–
EtOAc as the eluent to give 21 (2.82 g, 84%) as a syrup:
[a]D +29.1ꢁ (c 1.0, CHCl3); 1H NMR (400 MHz,
CDCl3): d 8.07–7.27 (m, 20H), 5.92 (t, 1H, J=9.8 Hz),
5.88 (m, 1H), 5.84 (d, 1H, J=3.0Hz), 5.69 (t, 1H,
J=9.8 Hz), 5.54 (dd, 1H, J=8.0, 9.6 Hz), 5.28 (dd, 1H),
5.17 (dd, 1H), 4.94 (d, 1H, J=8.0Hz), 4.67 (dd, 1H,
J=12.4, 3.2 Hz), 4.50(d, 1H, J=3.0Hz), 4.48 (dd, 1H,
J=12.4, 3.2 Hz), 4.12–3.86 (m, 8H), 1.39, 1.28 (2 s, 6H).
Anal. calcd for C46H46O15: C, 65.86; H, 5.53. Found: C,
65.39; H, 5.41.
ꢀ-D-Glucopyranosyl-(1!3)-[ꢀ-D-glucopyranosyl-(1!6)]-
ꢁ-D-glucopyranosyl-(1!3)-ꢀ-D-glucopyranosyl-(1!3)-
[ꢀ-D-glucopyranosyl-(1!6)]-D-glucopyranose (18). Com-
pound 17 (10g, 3.95 mmol) was dissolved in 80% acetic
acid solution (150mL) and the mixture was heated
under reflux for 5 h. The solution was concentrated, the
residue was purified by chromatography with 1.5:1 pet-
roleum ether–EtOAc as the eluent, and then the product
was treated with a saturated solution of ammonia in
CH2Cl2 (10mL) and CH 3OH (120mL) at rt. After 24 h,
the reaction mixture was concentrated, and the residue
was washed four times with CH2Cl2 to afford 18 as
1
white solid (3.6 g, 92%). [a]D ꢂ19.8 (c 0.1, H2O); H
NMR (400 MHz, D2O): d 5.27 (d, 1H, J=3.1 Hz), 4.74
(d, 1H, J=6.8 Hz), 4.65 (d, 1H, J=6.8 Hz), 4.64 (d, 1H,
J=6.8 Hz),4.44 (d, 1H, J=6.8 Hz),4.43 (d, 1H,
J=6.8 Hz); 13C NMR (100 MHz, D2O): d 102.69,
102.69, 102.6, 102.60, 102.60 (5 C-1, JC-H=163.9 Hz),
98.92 (C-1, JC-H=173.9 Hz), 85.34, 84.87, 82.9, 81.96.
Anal. calcd for C36H62O31: C, 43.64; H, 6.30. Found: C,
42.92; H, 6.89. ESMS for C36H62O31 (990.86): 989.84
[M-1]+.
2,3,4,6-Tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)-5-O-
benzoyl-1,2-O-isopropylidene-ꢁ-D-glucofuranose (23). To
a solution of 21 (4.8 g, 5.72 mmol) in pyridine (50mL)
was added BzCl (1.2 mL, 10mmol) at rt. After 2 h, the
mixture was poured to water and extracted with
CH2Cl2. The organic phase was concentrated, and the
resulting residue was treated with CH3OH (100 mL) and
PdCl2 (15 mg) at rt for 4 h, at the end of which time
TLC (2:1 petroleum ether–EtOAc) indicated that the
reaction was complete. The mixture was filtered, and the
filtrate was concentrated. The residue was passed a
silica-gel column with 2:1 petroleum ether–EtOAc as the
eluent to give 23 (4.0g, 78%) as a syrup; [ a]D +34.3ꢁ (c
Lauryl 2,3,4,6 - tetra - O - benzoyl - ꢀ - D - glucopyranosyl -
(1!3)-[2,3,4,6-tetra-O-benzoyl-ꢀ-D -glucopyranosyl-
(1!6)]-2,4-di-O-acetyl-ꢁ-D-glucopyranosyl-(1!3)-2,4,6-
tri-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-
benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-2,4-di-O-acetyl-ꢀ-D-
glucopyranoside (19). To a stirred solution of 9 (2.30g,
1.47 mmol) and 16 (1.75 g, 1.35 mmol) in CH2Cl2
(60mL) was added TMSOTf (30 mL) at rt. After 3 h, the
mixture was neutralized with triethylamine and con-
centrated. The residue was purified on a silica gel col-
umn with 1.5:1 petroleum ether–EtOAc as the eluent to
give 19 (3.1 g, 84%). [a]D +48.5 (c 1.7, CHCl3); 13C
NMR (100 MHz, CDCl3): d 170.47, 170.10, 169.62,
169.51, 169.50, 169.38, 168.76, 101.40, 101.28, 101.13,
100.65, 100.40, 93.16, 31.88–21.24, 20.75, 20.69, 20.65,
20.58, 20.48, 20.42, 14.09. Anal. calcd for C146H148O50:
C, 64.88; H, 5.52. Found: C, 64.43; H, 5.58.
1
1.0, CHCl3); H NMR (400 MHz, CDCl3): d 7.89–7.25
(m, 25H), 5.91 (d, 1H, J=3.0Hz), 5.87 (t, 1H,
J=9.8 Hz), 5.68 (t, 1H, J=9.8 Hz), 5.56 (dd, 1H,
J=8.1, 9.6 Hz), 5.31 (m, 1H), 5.05 (d, 1H, J=8.1 Hz),
4.62 (dd, 1H, J=12.1, 3.3 Hz), 4.55 (d, 1H, J=3.0Hz),
4.46 (dd, 1H, J=12.4, 3.3 Hz), 4.36–3.99 (m, 5H), 1.50,
1.31 (2 s, 6H). Anal. calcd for C50H46O16: C, 66.51; H,
5.13. Found: C, 66.11; H, 5.07.
2,3,4,6-Tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)-2,4-
di-O-acetyl-3-O-allyl-ꢁ-D-glucopyranosyl trichloroaceti-
midate (24). Compound 21 (12 g, 14.3 mmol) was added
to 80% aqueous acetic acid solution (150 mL) and the
mixture was heated under reflux for 5 h. The mixture
was concentrated and the residue was acetylated with
acetic anhydride (20mL) in pyridine (20mL) for 2 h at
rt. The resultant trisaccharide was dissolved in a 1.5 N
Lauryl ꢀ-D-Glucopyranosyl-(1!3)-[ꢀ-D-glucopyranosyl-
(1!6)]-ꢁ-D-glucopyranosyl-(1!3)-ꢀ-D-glucopyranosyl-
(1!3)-[ꢀ-D-glucopyranosyl-(1!6)]-ꢀ-D-glucopyranoside

2202
J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
solution of NH3 in 3:1 THF-CH3OH (100 mL), and the
solution was kept at rt. After 3 h, the solution was con-
centrated, and the residue was dissolved in CH2Cl2
(80mL). To the solution were added K 2CO3 (3,4 g,
25 mmol) and CCl3CN (2 mL, 20mmol), and the mix-
ture was stirred at rt for 24 h. Filtering the mixture, the
filtration and washings were concentrated, and the resi-
due was subjected to column chromatography to give 24
syl-(1!6)]-2,4-di-O-acetyl-ꢀ-D-glucopyransyl-(1!3)-
4,6-O-benzylidene-ꢀ-D-glucopyranoside (28). To a stir-
red solution of 9 (3.6 g, 2.3 mmol) and 2713 (865 mg,
2.3 mmol) in CH2Cl2 (30mL) was added TMSOTf
(30 mL) at room temperature. After 3 h, triethylamine
was added to the solution to quench the reaction. The
solution was concentrated, the residue was subjected to
column chromatography with 1.5:1 petroleum ether–
EtOAc as the eluent to give 28 (3.15 g, 77%): [a]D
ꢂ10.3 (c 1.0, CHCl3); H NMR (400 MHz, CDCl3): d
8.03–7.22 (m, 45H), 7.10, 6.83 (2 d, 4H, J=9.1 Hz), 5.89
(t, 1H, J=9.7 Hz), 5.73 ( t, 1H, J=9.4 Hz), 5.68–5.62
(m, 2H), 5.43–5.36 (m, 2H), 5.38 (s, 1H), 5.00 (t, 1H,
J=8.7 Hz ), 4.89 (d, 1H, J=7.6 Hz), 4.87 (dd, 1H,
J=7.8 Hz), 4.74 (d, 1H, J=7.9 Hz), 4.60(dd, 1H), 4.45
(m, 2H), 4.25–4.05 (m, 3H), 3.76 (s, 3H), 1.94, 1.91 (2 s,
6H); 13C NMR (100 MHz, CDCl3): d 169.88, 168.96,
166.15-164.80, 155.60, 151.26, 117.82, 114.68, 101.70,
101.10, 100.70, 100.38. Anal. calcd for C98H88O32: C,
66.21; H, 4.99. Found: C, 66.64; H, 4.89.
(11 g, 75%) as a syrup: [a]D +42.4ꢁ (c 1.0, CHCl3); H
1
ꢁ
1
NMR (400 MHz, CDCl3): d 8.35 (s, 1H), 8.04–7.26 (m,
20H), 6.32 (d, 1H, J=3.2 Hz), 5.86 (t, 1H, J=9.6 Hz),
5.75 (m, 1H), 5.64 (t, 1H, J=9.8 Hz), 5.48 (dd, 1H,
J=8.0, 9.6 Hz), 5.16 (dd, 1H), 5.10 (dd, 1H), 4.96 (d,
1H, J=8.0Hz), 4.86 (t, 1H, J=9.8 Hz), 4.84 (dd, 1H,
J=10.4, 3.0 Hz), 4.61 (dd, 1H, J=10.4, 3.2 Hz), 4.49
(dd, 1H, J=9.6, 3.2 Hz), 4.17–3.90(m, 5H), 3.86 (t, 1H),
3.71 (dd, 1H), 2.04, 2.00 (2 s, 6H). Anal. calcd for
C49H46NO17Cl3: C, 57.29; H, 4.51. Found: C, 57.71; H,
4.35.
Lauryl 2,3,4,6 - tetra - O - benzoyl - ꢀ - D - glucopyranosyl -
(1!6)-2,4-di-O-acetyl-3-O-allyl-ꢀ-D-glucopyranoside
(25). To a solution of 24 (2.5 g, 2.43 mmol) and lauryl
alcohol (744 mg, 4.0mmol) in CH 2Cl2 (50mL) was
added TMSOTf (30 mL) at rt. The reaction mixture was
stirred for 3 h, at the end of which time TLC indicated
that the reaction was complete. Then the mixture was
neutralized with triethylamine and concentrated under
reduced pressure to dryness. Purification by column
chromatography (3:1 petroleum ether–EtOAc) gave 25
2,3,4,6-Tetra-O-benzoyl-ꢀ-D -glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)-]2,4-
di-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-2,4,6-tri-O-ace-
tyl-ꢁ-D-glucopyranosyl trichloroacetimidate (29).
A
solution of 28 (5 g, 2.81 mmol) in 90% acetic acid
ꢁ
(80mL) was kept at 40 C for 24 h and then con-
centrated to a residue under reduced pressure. The resi-
due was treated with pyridine (10mL) and Ac 2O
(10mL) at rt for 2 h. This mixture was added to water
and extracted with CH2Cl2. The organic phase was
dried over Na2SO4, and concentrated, the residue was
dissolved in 4:1 CH3CN–H2O (40mL). To this solution
was added (NH4)2Ce(NO3)6 (2.72 g, 5.0mmol), and the
mixture was stirred for 30min at rt, at the end of which
time TLC (1:1 petroleum ether–EtOAc) indicated that
the reaction was complete. The mixture was extracted
with CH2Cl2, and washed with satd aq NaHCO3. The
organic layer was concentrated, the residue was dis-
solved in dichloromethane. To the solution were added
K2CO3 (1.4 g, 10mmol) and CCl 3CN (0.5 mL, 5 mmol),
and the mixture was stirred at rt for 24 h. Filtering the
mixture, the filtration and washings were concentrated,
and the residue was subjected to column chromato-
graphy (1.5:1 petrꢁoleum ether–EtOAc) to give 29 (3.7 g,
as a syrup (2.35 g, 92%):[a]D +33.70 (c 1.0, CHCl3); H
1
NMR (400 MHz, CDCl3): d 8.04–7.27 (m, 20H), 5.88 (t,
1H, J=9.7 Hz), 5.71 (m, 1H), 5.66 (t, 1H, J=9.6 Hz),
5.51 (dd, 1H, J=8.0, 9.6 Hz), 5.16 (dd, 1H), 5.10 (dd,
1H), 4.95 (d, 1H, J=8.0Hz), 4.85 (t, 1H, J=9.8 Hz),
4.78 (t, 1H, J=9.8 Hz), 4.63 (dd, 1H, J=10.4, 2.0 Hz),
4.47 (dd, 1H, J=10.4, 3.2 Hz), 4.19 (d, 1H, J=9.4 Hz),
4.15 (m, 1H), 4.01 (d, 2H), 3.88 (d, 1H), 3.65 (t, 1H),
3.60–3.40 (m, 3H), 3.05 (m, 1H), 2.04, 2.00 (2 s, 6H),
1.33–1.12 (m, 20H), 0.88 (t, 3H). Anal. calcd for
C59H70O17: C, 67.41; H, 6.71. Found: C, 67.02; H, 6.59.
Lauryl 2,3,4,6 - tetra - O - benzoyl - ꢀ - D - glucopyranosyl -
(1!6)-2,4-di-O-acetyl-ꢀ-D-glucopyranoside (26). To a
solution of 25 (3.4 g, 3.23 mmol) in CH3OH (50mL) was
added PdCl2 (20mg), and the mixture was stirred for 4 h
at rt, at the end of which time TLC (2:1 petroleum
ether–EtOAc) indicated that the reaction was complete.
The mixture was filtered, the filtrate was concentrated,
and the residue was passed a silica-gel column with 2:1
petroleum ether–EtOAc as the eluent to give 26 as a
syrup (2.71 g, 83%): [a]D +14.9ꢁ (c 1.0, CHCl3); 1H
NMR (400 MHz, CDCl3): d 8.05–7.25 (m, 20H), 5.89 (t,
1H, J=9.6 Hz), 5.68 (t, 1H, J=9.6 Hz), 5.52 (dd, 1H,
J=7.8, 9.6 Hz), 4.95 (d, 1H, J=7.8 Hz), 4.75–4.67 (m,
3H), 4.47 (dd, 1H, J=4.9, 12.1 Hz), 4.24 (d, 1H,
J=7.8 Hz), 4.15 (m, 1H), 3.92 (dd, 1H), 3.72–3.50(m,
4H), 3.12 (m, 1H), 2.07, 2.02 (2 s, 6H), 1.38–1.11 (m,
20H), 0.88 (t, 3H, J=6.6 Hz). Anal. calcd for
C56H66O17: C, 66.52; H, 6.58. Found: C, 66.05; H, 6.74.
1
71%): [a]D +4.0 (c 1.0, CHCl3); H NMR (400 MHz,
CDCl3): d 8.65 (s, 1H), 8.15–7.24 (m, 40H), 6.38 (d, 1H,
J=3.5 Hz), 5.88 (t, 1H, J=9.4 Hz), 5.87 (t, 1H,
J=9.4 Hz), 5.66 (t, 1H, J=9.4 Hz), 5.64 (t, 1H,
J=9.3 Hz), 5.50(dd, 1H, J=7.8, 9.0Hz), 5.37 (dd, 1H,
J=7.8, 9.0Hz), 4.92–4.83 (m, 4H), 4.74–4.63 (m, 3H),
4.60–4.45 (m, 3H), 4.40 (d, 1H, J=8.1 Hz), 4.16–3.56
(m, 10H), 2.08, 2.02, 1.86, 1.81, 1.74 (5 s, 15H); 13C
NMR (100 MHz, CDCl3): d 170.77, 169.73, 169.71,
169.01, 168.09, 160.99, 101.32, 100.79, 100.66, 93.23,
20.72, 20.68, 20.60, 20.50, 20.32. Anal. calcd for
C92H84NO34Cl3: C, 59.60; H, 4.57. Found: C, 59.09; H,
4.63.
Lauryl 2,3,4,6 - tetra - O - benzoyl - ꢀ - D - glucopyranosyl -
(1!3)-[2,3,4,6-tetra-O-benzoyl-ꢀ-D -glucopyranosyl-
(1!6)]-2,4-di-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-2,4,6-
tri-O-acetyl-ꢀ-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-
4-Methoxyphenyl 2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyr-
anosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyrano-

J. Ning et al. / Bioorg. Med. Chem. 11 (2003) 2193–2203
2203
benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-2,4-di-O-acetyl-ꢀ-D-
glucopyranoside (30) and Lauryl 2,3,4,6-tetra-O-benzoyl-
ꢀ-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-ꢀ-D-
glucopyranosyl-(1!6)]-2,4-di-O-acetyl-ꢀ-D-glucopyrano-
syl-(1!3)-2,4,6-tri-O-acetyl-ꢁ-D-glucopyranosyl-(1!3)-
[2,3,4,6-tetra-O-benzoyl-ꢀ-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-ꢀ-D-glucopyranoside (31). To a stirred solu-
tion of 29 (3.6 g, 1.94 mmol) and 26 (1.82 g, 1.8 mmol) in
CH2Cl2 (30mL) was added TMSOTf (20 mL) at room
temperature. After 3 h, triethylamine was added to the
solution to quench the reaction. The solution was con-
centrated, the residue was subjected to column chroma-
tography with 1:1 petroleum ether–EtOAc as the eluent
to give 30 (1.85 mg, 38%) and 31 (838 mg, 16%): For
30: [a]D +38ꢁ (c 1.0, CHCl3); 13C NMR (100 MHz,
CDCl3): d 170.35, 169.52, 169.38, 168.82, 168.63,
168.63, 168.28, 101.18, 100.18, 100.28, 100.28, 100.19,
100.19 (6 C-1, JC-H=163.2-163.3 Hz), 78.45, 77.96,
77.94, 20.71, 20.65, 20.55, 20.47, 20.38, 20.30, 20.25.
Anal. calcd for C146H148O50: C, 64.88; H, 5.52. Found:
C, 64.40; H, 5.65; For 31: [a]D +8.5ꢁ (c 1.0, CHCl3);
13C NMR (100 MHz, CDCl3): d 170.52, 170.27, 169.67,
169.29, 169.06, 168.87, 167.88, 101.15, 101.08, 100.84,
100.54, 100.54, 95.19, 20.91, 20.80, 20.79, 20.65, 20.48,
40.30, 20.30. Anal. calcd for C146H148O50: C, 64.88; H,
5.52. Found: C, 64.56; H, 5.47.
(33). Compound 31 (470mg, 0.17 mmol) was dissolved
in a solution of CH2Cl2 (5 mL) and CH3OH (50mL)
saturated with NH3 at rt. After 24 h, the reaction mix-
ture was concentrated, and the residue was washed four
times with CH2Cl2 to afford 33 as white solid (183 mg,
93%). [a]D ꢂ12.1ꢁ (c 1.0, H2O); 13C NMR (100 MHz,
CDCl3): d 102.70, 102.68, 102.68, 102.65, 102.65 (5 C-1,
JC-H=164.2-164.9 Hz ), 99.15 (C-1, JC-H=175.44 Hz),
84.24, 83.47, 82.23. Anal. calcd for C48H86O31: C, 49.74;
H, 7.48. Found: C, 49.17; H, 7.89. ESMS for C48H86O31
(1159.18): 1158.17 [M-1]+.
Acknowledgements
This work was supported by the Beijing Natural
Science Foundation (6021004) and the National Nat-
ural Science Foundation of China (Project 59973026
and 29905004), and by The Ministry of Science and
Technology.
References and Notes
1. Sasaki, T.; Takasuka, N. Carbohydr. Res. 1976, 47, 99.
2. Oka, M.; Hazama, S.; Suzuki, M.; Wang, F.; Wadamori,
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K.; Suga, T.; Suzuki, T. Int. J. Immunopharmocology 1996, 18,
211.
3. (a) Norisue, T.; Yanaki, T.; Fujita, H. J. Polym. Sci. 1980,
18, 547. (b) Kojima, T.; Tabata, K.; Itoh, W.; Yanaki, T.
Agric. Biol. Chem. 1986, 50, 231. (c) Brown, G. D.; Gordon, S.
Nature 2001, 413, 36. (d) Willment, J. A.; Gordon, S.; Brown,
G. D. J. Biol. Chem. 2001, 276, 43818. (e) Mueller, A.; Raptis,
J.; Rice, P. J.; Kalbfleisch, J. H.; Stout, R. D.; Ensley, H. E.;
Browder, W.; Williams, D. L. Glycobiology 2000, 10, 339.
4. Sasaki, T.; Takasuka, N.; Chihara, G.; Maeda, Y. Y. Gann.
1976, 67, 191.
Orthoester 300. To a stirred solution of 29 (1.30g,
0.7 mmol) and 26 (0.71 g, 0.7 mmol) in CH2Cl2 (20mL)
was added TMSOTf (10 mL) at room temperature. After
20min, triethylamine was added to the solution to
quench the reaction. The solution was concentrated, the
residue was subjected to column chromatography with
1.5:1 petroleum ether–EtOAc as the eluent to give 300
(1.32 g, 71%): [a]D +17.6ꢁ (c 1.0, CHCl3); 13C NMR
(100 MHz, CDCl3): d 170.53, 169.57, 169.46, 169.46,
169.08, 167.92, 121.90, 101.20, 101.04, 100.49, 100.38,
100.28, 96.60. Anal. calcd for C146H148O50: C, 64.88; H,
5.52. Found: C, 65.22; H, 5.67.
5. Sharp, J. K.; Valent, B.; Albersheim, P. J. Biol. Chem.
1984, 259, 11312.
6. Toshima, K.; Tatsuta, K. Chem. Rev. 1993, 93, 1500.
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99119757.7, 1999. (b) Ning, J.; Kong, F. PCT WO 01/23397
A1.
8. Ning, J.; Yuetao, Y.; Kong, F. Tetrahedron Lett. 2002, 43,
5545.
9. (a) Ning, J.; Kong, F. Chinese Patent, Application No:
99126224.7, 1999. (b) Ning, J.; Kong, F. Chinese Patent,
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00100376.3, 2000. (b) Ning, J.; Kong, F. PCT WO 01/44263
A1.
Lauryl
ꢀ-D-glucopyranosyl-(1!3)-[ꢀ-D-glucopyrano-
syl(1!6)]-ꢀ-D-glucopyranosyl-(1!3)-ꢀ-D-glucopyranosyl-
(1!3)-[ꢀ-D-glucopyranosyl-(1!6)]-ꢀ-D-glucopyranoside
(32). Compound 30 (1.2 g, 0.44 mmol) was dissolved in a
saturated solution of NH3 in CH2Cl2 (5 mL) and
CH3OH (50mL) at rt. After 24 h, the reaction mixture
was concentrated, and the residue was washed four
times with CH2Cl2 to afford 32 as white solid
(470mg, 92%): [ a]D ꢂ8.4ꢁ (c 1.0, H2O); 1H NMR
(400 MHz, D2O): d 4.39–3.98 (m, 6H), 3.85–3.09 (m,
38H), 1.51 (m, 2H), 1.22–1.12 (m, 18H,), 0.76 (t, 3H);
11. Zeng, Y.; Ning, J.; Kong, F. Tetrahedron Lett. 2002, 43,
3279.
12. (a) Verduyn, R.; Douwes, M.; van der Klein, P. A. M.;
13C NMR (100 MHz, CDCl3):
d 105.52, 105.33,
105.23, 105.18, 105.14, 104.94 (6 C-1), 88.16, 87.42,
86.81. Anal. calcd for C48H86O31: C, 49.74; H, 7.48.
Found: C, 49.07; H, 7.76. ESMS for C48H86O31
(1159.18): 1158.16 [M-1]+.
Mosinger, E. M.; van der Marel, G. A.; van Boom, J. H. Tet-
¨
rahedron 1993, 49, 7301. (b) Takeo, K.; Maki, K.; Wada, Y.;
Kitamura, S. Carbohydr. Res. 1993, 245, 81.
13. Yang, G.; Kong, F. Synlett. 2000, 1423.
14. (a) Chen, L.; Kong, F. J. Carbohydr. Chem. 2002, 21, 341. (b)
Ogawa, T.; Yamamoto, H. Agric. Biol. Chem. 1985, 49, 475.
15. Schmidt, R. R.; Kinzy, W. Adv. Carbohydr. Chem. Bio-
chem. 1994, 50, 21.
Lauryl ꢀ-D -glucopyranosyl-(1!3)-[ꢀ-D -glucopyranosyl
(1!6)]-ꢀ-D-glucopyranosyl-(1!3)-ꢁ-D-glucopyranosyl-
(1!3)-[ꢀ-D-glucopyranosyl-(1!6)]-ꢀ-D-glucopyranoside