Synthesis of β-D-Glcp-(1→3)-[β-D-Glcp-(1→6)]-β-D- Glcp-(1→3)-β-D-Glcp-(1→6)-[β-D-Galp-(1→4) -β-D-Glcp-(1→3)]-β-D-GlcpOLauryl, an oligosaccharide with anti-tumor activity

Article abstract of DOI:10.1016/j.carres.2005.08.010

A concise and effective synthesis of lauryl heptasaccharide 17 was achieved from the key intermediates lauryl 2,3,4,6-tetra-O-benzoyl-β-D- galactopyranosyl-(1→4)-2,3,6-tri-O-benzoyl-β-D-glucopyranosyl- (1→3)-2,4-di-O-benzoyl-β-D-glucopyranoside (10) and i

Full text of DOI:10.1016/j.carres.2005.08.010

Carbohydrate
RESEARCH
Carbohydrate Research 340 (2005) 2345–2351
Synthesis of b-D-Glcp-(1!3)-[b-D-Glcp-(1!6)]-
b-D-Glcp-(1!3)-b-D-Glcp-(1!6)-[b-D-Galp-(1!4)-b-D-Glcp-(1!3)]-
b-D-GlcpOLauryl, an oligosaccharide with anti-tumor activity
Xiangdong Mei,b Linsen Heng,a,* Mingkun Fu,b Zhimin Lia and Jun Ningb,*
aCollege of Bio-information, Chongqing University of Post and Telecommunications, Chongqing 400065, PRChina
bResearch Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, PR China
Received 1 August 2005; received in revised form 21 August 2005; accepted 24 August 2005
Abstract—A concise and effective synthesis of lauryl heptasaccharide 17 was achieved from the key intermediates lauryl 2,3,4,6-
tetra-O-benzoyl-b-D-galactopyranosyl-(1!4)-2,3,6-tri-O-benzoyl-b-D-glucopyranosyl-(1!3)-2,4-di-O-benzoyl-b-D-glucopyranoside
(10) and isopropyl 2,4,6-tri-O-acetyl-3-O-allyl-b-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glucopyranosyl-(1!6)]-2,4-
di-O-acetyl-b-D-glucopyranosyl-(1!3)-2,4,6-tri-O-acetyl-1-thio-b-D-glucopyranoside (15). The key trisaccharide glycosyl acceptor
10 was constructed by coupling 2,3,4,6-tetra-O-benzoyl-b-D-galactopyranosyl-(1!4)-2,3,6-tri-O-benzoyl-a-D-glucopyranosyl tri-
chloroacetimidate (3) with lauryl 6-O-acetyl-2,4-di-O-benzoyl-b-D-glucopyranoside (9), followed by deacetylation. The thioglycoside
donor 15 was obtained by condensation of 2,4,6-tri-O-acetyl-3-O-allyl-b-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-
glucopyranosyl-(1!6)]-2,4-di-O-acetyl-a-D-glucopyranosyl trichloroacetimidate (11) with isopropyl 4,6-O-benzylidene-1-thio-b-D-
glucopyranoside (12), followed by debenzylidenation and acetylation. A bioassay of the inhibition of S180 noumenal tumors showed
that lauryl heptasaccharide 17 could be employed as a potential agent for cancer treatment.
ꢀ 2005 Elsevier Ltd. All rights reserved.
Keywords: Oligosaccharide; Glycosylation; Anti-tumor
OH
O
1. Introduction
OH
O
OH
HO
O
HO
OH
OH
Evidence has accumulated proving that simple sugars
can be employed to inhibit the aggregation of certain
types of tumor cells.1,2 For example, Raz and Lotan3,4
found that the lectin-like activity of several tumor cell
lines, including five human melanomas (A375, SH4,
Hs294, Hs852, and Hs939), human cervical adenocarci-
noma (HeLa-S3), murine melanoma (B16-F1), and mur-
ine fibrosarcoma (UV-2237P) could be inhibited by
lactose (1, Fig. 1). Accordingly, it is reasonable to be-
lieve that the structure of lactose (4-O-b-D-galactopyr-
anosyl-D-glucopyranose) may be useful for finding new
OH
1
OH
O
HO
O
HO
OH
OH
O
OH
O
HO
O
O
HO
O
HO
OH
OH
OH
n
2
Figure 1. Structures of lactose 1 and schizophyllan 2.
drugs to inhibit the lectin-like ability of tumor cells
that play an important role in tumor growth at the
primary site, invasion into surrounding host tissue,
*
Corresponding authors. Tel.: +86 10 62849157; fax: +86 10 62923563
(J.N.); e-mail: jning@mail.rcees.ac.cn
0008-6215/$ - see front matter ꢀ 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2005.08.010

2346
X. Mei et al. / Carbohydrate Research 340 (2005) 2345–2351
dissemination, embolization, and implantation at dis-
tant secondary sites to form metastases.
2. Results and discussion
Schizophyllan (2), produced by Schizophyllum com-
mune Fries, consists of a main chain of (1!3)-b-linked
D-glucose residues with one (1!6)-b-linked D-glucopyr-
anosyl group for every three glucopyranose residues
(Fig. 1).5,6 Investigations on the application of schizo-
phyllan as an anti-tumor or anti-infective agent showed
that schizophyllan appears to have a curative effect
against Sarcoma-180 ascites, lung cancer, stomach can-
cer, and ovarian cancer.7–14
In our synthesis, 2,3,4,6-tetra-O-benzoyl-b-D-galact-
opyranosyl-(1!4)-2,3,6-tri-O-benzoyl-a-D-glucopyran-
osyl trichloroacetimidate (3) and lauryl 6-O-acetyl-2,4-di-
O-benzoyl-b-D-glucopyranoside (9) were the starting
materials; trisaccharide 10 and the tetrasaccharide 15
were the key intermediates. Compound 3 was prepared
as fine crystals via benzoylation of D-lactose (1) followed
by 1-O-debenzoylation with ammonia in tetrahydrofu-
ran–methanol and subsequent treatment with trichloro-
acetonitrile in the presence of potassium carbonate26
(Scheme 1). Tritylation of 3-O-allyl-D-glucopyranose
(4)27 followed by benzoylation in a one-pot manner gave
3-O-allyl-1,2,4-tri-O-benzoyl-6-O-triphenylmethyl-b-D-
glucopyranose (5, 71%), detritylation of which afforded
3-O-allyl-1,2,4-tri-O-benzoyl-b-D-glucopyranose (6) in
90% yield. Acetylation of 6 with acetic anhydride in pyr-
idine, then selective 1-O-debenzoylation with ammonia in
tetrahydrofuran–methanol followed by treatment with
trichloroacetonitrile in the presence of potassium carbon-
ate in dichloromethane, afforded 6-O-acetyl-3-O-allyl-
2,4-di-O-benzoyl-a-D-glucopyranosyl trichloroacetimi-
date (8) in 77% yield (over the three steps). Coupling of
8 with lauryl alcohol catalyzed by trimethylsilyl trifluo-
romethanesulfonate (TMSOTf) in dry dichloromethane,
followed by 3-O-deallylation using palladium chloride
in methanol–dichloromethane, afforded the glycosyl
acceptor lauryl 6-O-acetyl-2,4-di-O-benzoyl-b-D-gluco-
pyranoside (9) in 78% overall yield (over two steps).
Although numerous carbohydrate structures occur in
nature, in general, the role of oligosaccharide structures
has not been widely studied. This can be attributed
mainly to difficulties in synthesizing oligosaccharides.
Unlike proteins and nucleic acids, oligosaccharides are
more difficult to synthesize because (i) the molecules
are typically branched rather than linear, (ii) the mono-
saccharide units can be connected by a or b linkages,
and (iii) oligosaccharide synthesis requires multiple
selective protection and deprotection steps. Although
over the past fewdecades, considerable progress has
been made in this field,15 there is still no general route
for oligosaccharide synthesis and glycosylation chemis-
try is often not predictable. To facilitate the synthesis
of target oligosaccharides, regio- and stereoselective gly-
cosylation of glycosyl donors with unprotected or par-
tially protected sugar acceptors has been extensively
studied. Employing this strategy, in the last fewyears,
we have prepared many oligosaccharides with various
structures such as 3,6-branched gluco-oligosaccha-
rides,16 2,6-branched manno-oligosaccharides,17 as well
as 2,6-branched,18 3,6-branched, and 5,6-branched
galacto-oligosaccharides.19
Several approaches have been taken with success for
the chemical synthesis of oligosaccharides.2 Most in-
volve the activation of the anomeric leaving group with
a Lewis acid and then displacement of that leaving
group by the free hydroxyl of the acceptor sugar. The
Koenigs–Knorr method for coupling glycosyl halides,
one of the first techniques to gain widespread usage, is
still in common use. Trichloroacetimidates,20 prepared
by the reaction of reducing sugars with trichloroaceto-
nitrile and a base are used frequently for coupling, as
are glycosyl sulfoxides,21 phosphates22 and phos-
phates,23 and thio-24 and pentenyl glycosides.25
OBz
O
OH
O
OBz
OBz
OH
O
OH
HO
a
O
O
O
BzO
HO
BzO
OH
BzO
OH
OBz
OC(NH)CCl3
OH
3
1
OH
O
OTr
b
c
O
OBz
HO
AllO
BzO
AllO
OH
OH
4
OBz
5
OH
O
OAc
O
d
e
OBz
OBz
BzO
AllO
BzO
AllO
OBz
7
OBz
6
OAc
O
OAc
O
f
OC12H25
BzO
AllO
BzO
HO
OBz
BzO
Providing sufficient quantities of a sample is a basic
prerequisite for detailed studies on a compoundÕs
fundamental biochemical properties and possible bio-
logical functions. To further study the structure–activ-
ity relationships of lactose and schizophyllan, we
report here the synthesis of compound 17, which con-
tains a glucotetraose moiety, the key fraction of schizo-
phyllan, and lactose. We anticipated that these two
structural motifs would be able to amplify anti-tumor
activity.
OC(NH)CCl3
9
8
Scheme 1. Reagents and conditions: (a) (i) BzCl, pyridine, rt, 12 h; (ii)
3:2 (v/v), THF/CH3OH saturated dry NH3, rt, 3 h; (iii) CH2Cl2,
CCl3CN and K2CO3, rt, 12 h, 56% (over three steps); (b) (i)
chlorotriphenylmethane (1.2 equiv), pyridine, 40 ꢁC, 48 h; (ii) BzCl
(3.3 equiv), rt, 24 h, 71% (for two steps); (c) CH2Cl2/CH3OH/0.1%
AcCl, rt, 1 h, 90%; (d) Ac2O/pyridine, rt, 5 h; (e) (i) 3:2 (v/v), THF/
CH3OH saturated dry NH3, rt, 3 h; (ii) CH2Cl2, CCl3CN and K2CO3,
rt, 12 h, 77% from 6; (f) (i) lauryl alcohol, TMSOTf, CH2Cl2, rt, 2 h;
(ii) 2:1 (v/v), CH3OH/CH2Cl2, PdCl2, rt, 6 h, 78% (for two steps).

X. Mei et al. / Carbohydrate Research 340 (2005) 2345–2351
2347
With the building blocks 3, 9, 2,4,6-tri-O-acetyl-3-O-al-
lyl-b-D-glucopyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-
b-D-glucopyranosyl-(1!6)]-2,4-di-O-acetyl-a-D-gluco-
pyranosyl trichloroacetimidate (11),27 and isopropyl
4,6-O-benzylidene-1-thio-b-D-glucopyranoside (12)28 in
hand, construction of the target compound was readily
carried out. As shown in Scheme 2, condensation of 3
and 9 catalyzed by trimethylsilyl trifluoromethanesulfo-
nate in dry dichloromethane, followed by deacetylation
with acetyl chloride in methanol–dichloromethane29 gave
the corresponding trisaccharide 10. Regio- and stereose-
lective coupling27,28,30 of 11 with 12 followed by debenz-
ylidenation in 90% acetic acid and acetylation with
acetic anhydride in pyridine afforded the desired tetrasac-
charide thioglycoside donor 15 in 92% yield (over three
steps). Using 10 as the glycosyl acceptor and 15 as the gly-
cosyl donor, the fully protected lauryl heptasaccharide 16
was easily obtained using N-iodosuccinimide (NIS) and
trimethylsilyl trifluoromethanesulfonate as catalysts in
dichloromethane. Deallylation of 16 using palladium
OH
O
OBz
OBz
OBz
O
a
OC12H25
O
BzO
+
3
9
O
O
BzO
BzO
OBz
OBz
OBz
10
OBz
O
O
Ph
BzO
BzO
O
O
O
+
SCH(CH3)2
OBz
HO
O
OAc
OH
AcO
O
O
AcO
OC(NH)CCl3
AllO
AcO
OAc
12
11
OBz
O
BzO
AllO
O
BzO
b
c
OBz
O AcO
Ph
O
O
O
OAc
O
SCH(CH3)2
SCH(CH3)2
O
O
OH
AcO
OAc
OAc
13
OBz
O
BzO
BzO
O
OR
OBz
O
OAc
O AcO
RO
O
O
O
AcO
OR
AllO
OAc
OAc
14 R = H
d
15 R = Ac
OBz
O
BzO
BzO
O
OAc
OBz
e
O
OAc
+
15
10
O
O
AcO
O
O
AcO
O
AcO
OAc
OBz
AllO
OAc
OAc
OBz
OBz
O
O
OC12H25
O
BzO
O
O
BzO
OBz
BzO
OBz
OBz
16
OH
O
HO
HO
O
HO
OH
OH
OH
f
O
O
O
HO
O
O
HO
O
HO
OH
OH
OH
OH
O
OH
OH
HO
O
OC12H25
HO
O
O
O
HO
OH
OH
OH
17
Scheme 2. Reagents and conditions: (a) (i) TMSOTf, CH2Cl2, rt, 2 h; (ii) CH3OH/CH2Cl2, AcCl, rt, 7 h, 85% (for two steps); (b) TMSOTf, CH2Cl2,
rt, 2 h; (c) 90% HOAc/H2O, reflux, 2 h; (d) Ac2O/pyridine, rt, 5 h, 92% from 11 and 12; (e) CH2Cl2, NIS, TMSOTf, rt, 2 h, 87%; (f) (i) CH3OH/
CH2Cl2, PdCl2, rt, 6 h; (ii) CH3OH/CH2Cl2 saturated with NH3, rt, 96 h, 82% (for two steps).

2348
X. Mei et al. / Carbohydrate Research 340 (2005) 2345–2351
Table 1. Inhibition of S180 tumor by 17 and CPA
Groups
Number of mice
Weight (g)
Tumor weight (g)
Inhibition (%)
S180 Cell line group
20
10
10
10
10
28.59 0.95
29.73 1.00
29.83 0.90
29.41 0.82
27.30 0.69
1.69 0.64
Group 1: treatment with 17 (1 mg/kg)
Group 2: treatment with 17 (2 mg/kg)
Group 3: treatment with 17 (4 mg/kg)
Group 4: treatment with CPA (40 mg/kg)
1.03 0.63*
1.02 0.52*
1.05 0.32*
0.29 0.13**
39.1
39.9
37.9
82.8
Compared to S180 cell line group, *p < 0.01, **p < 0.001.
chloride in methanol–dichloromethane, followed by
deprotection with ammonia in methanol, gave the target
compound 17 in excellent (82%) yield.
at <60 ꢁC under reduced pressure. Dry reaction solvents
were distilled over CaH2 and stored over molecular
sieves.
Inhibition of S180 noumenal tumors by the synthetic
oligosaccharide was investigated. The tumor inhibition
ratios for 17 were 39.1%, 39.9%, and 37.9% at a dose
of 1, 2, and 4 mg/kg/day, respectively (Table 1). These
activities can be compared with cyclophosphamide
(CPA), which gave 82.8% tumor inhibition at a concen-
tration of 40 mg/kg/day. These results indicate that 17 is
a potent inhibitor of tumor growth and thus potential
agent for cancer treatment.
3.2. Bioassay for inhibition of S180
CPA was purchased from Shanghai Hualian Pharma-
ceutical Co. Ltd (No. 030907). Sixty female and male
Kunming mice (weight 18–22 g, grade SPF) were ob-
tained from the Chongqing Academy of Chinese Mate-
ria Medica. The S180 cell line was provided by the
Department of Tumorology, Institute of Pharmacy,
the Chinese Academy of Medicine. The mice were ran-
domly divided into five groups according to weight,
and the S180 cell line of ascites cancer 0.2 mL (amount
of cell 2 · 106) was planted intraperitoneally into the
euterocelia. At 24 h after the planting, the mice were
treated with 17 or CPA. The oligosaccharide 17 was dis-
solved in saline at 0.1 mg/mL, given at a dose of 1, 2, or
4 mg/kg/day to the mice for nine consecutive days. CPA
was dissolved in saline at 4 mg/mL and administered
intraperitoneally to the mice at a dose of 40 mg/kg/
day over nine consecutive days.
In conclusion, a highly efficient and concise synthesis
of 17, a heptasaccharide with anti-tumor activity, was
achieved through the regio- and stereoselective glycosyl-
ations. All newcompounds involved in this study were
1
characterized by optical rotations, H NMR and 13C
NMR spectroscopy, and elemental analyses or electro-
spray-ionization MS. In all reactions, easily accessible
materials and inexpensive reagents were used and the
reactions were carried out in high yields and in large
scale. In executing the synthesis, several intermediates
were not separated but instead were used directly in
the next reaction thus substantially streamlining the
synthesis.
3.3. 2,3,4,6-Tetra-O-benzoyl-b-D-galactopyranosyl-
(1!4)-2,3,6-tri-O-benzoyl-a-D-glucopyranosyl
trichloroacetimidate (3)
3. Experimental
D-Lactose (1, 3.6 g, 10.0 mmol) was treated with BzCl
(11.2 mL, 96.0 mmol) and pyridine (40 mL) for 12 h at
rt. The benzoylated sugar was dissolved in a solution
of 3:2 THF–CH3OH (120 mL) and saturated dry NH3.
The solution was kept at rt for 3 h at the end of which
time TLC (1:1, petroleum ether–EtOAc) indicated that
the reaction was complete. The mixture was concen-
trated and the residue was dissolved in a solution of
CH2Cl2 (60 mL) and CCl3CN (1.5 mL, 15.0 mmol) con-
taining K2CO3 (4.1 g, 30.0 mmol). The reaction mixture
was stirred for 12 h at rt. The mixture was filtered and
the combined filtrate and washings were concentrated,
and the residue was purified by column chromatography
(3:1, petroleum ether–EtOAc) to give 3 (6.8 g, 56% for
three steps) as a white solid: [a]D +54.5 (c 1.0, CHCl3);
3.1. General methods
Melting points were determined with a Mel-Temp appa-
ratus. Optical rotations were measured at 25 ꢁC in the
stated solvent and are in units of degrees mL/g dm. H
1
NMR (400 MHz) and 13C NMR (100 MHz) spectra
were recorded in CDCl3 solutions at room temperature
unless otherwise specified. Chemical shift (d) values are
given in ppm; coupling constants (J) are in hertz. Mass
spectra were recorded on an Autospec mass spectro-
meter using the electrospray-ionization technique. Thin-
layer chromatography (TLC) was performed on silica
gel HF254 with detection by charring with 30% (v/v)
H2SO4 in CH3OH or in some cases with a UV detector.
Column chromatography was conducted by elution of a
column (10 · 240 mm, 18 · 300 mm od 35 · 400 mm) of
silica gel (100–200 mesh) with EtOAc–petroleum ether
(60–90 ꢁC) as the eluent. Solutions were concentrated
1H NMR (CDCl3, 400 MHz):
d
8.56 (s, 1H,
OC(NH)CCl3), 7.99–7.19 (m, 35H, BzH), 6.71 (d, 1H,
J = 3.6 Hz, H-1), 6.16 (t, 1H, J = 9.9 Hz, H-3), 5.77–
5.72 (m, 2H, H-20, H-40), 5.54 (dd, 1H, J = 3.6,

X. Mei et al. / Carbohydrate Research 340 (2005) 2345–2351
2349
10.2 Hz, H-2), 5.38 (dd, 1H, J = 3.3, 10.3 Hz, H-30), 4.93
3.5. Lauryl 6-O-acetyl-2,4-di-O-benzoyl-b-D-gluco-
pyranoside (9)
(d, 1H, J = 7.9 Hz, H-10), 4.58–4.50 (m, 2H, H-6a, 6b),
4.36–4.32 (m, 2H, H-4, H-5), 3.90 (m, 1H, H-50), 3.74–
3.69 (m, 2H, H-6a0, 6b0). Anal. Calcd for
C63H50Cl3NO18: C, 62.26; H, 4.15. Found: C, 62.15;
H, 4.32.
Trimethylsilyl trifluoromethanesulfonate (50 lL) was
added to a solution of 8 (3.07 g, 5.0 mmol) and lauryl
alcohol (1.12 g, 6.0 mmol) in CH2Cl2 (50 mL) at rt.
The reaction mixture was stirred for 2 h, at the end of
which time TLC (5:1, petroleum ether–EtOAc) indicated
that the reaction was complete. The mixture was neu-
tralized with Et3N and concentrated. The resulting resi-
due was dissolved in a solution of CH3OH (40 mL) and
CH2Cl2 (20 mL). To this solution was added PdCl2
(60 mg), and the mixture was stirred for 6 h at rt, at
the end of the which time TLC (4:1, petroleum ether–
EtOAc) indicated that the reaction was complete. The
mixture was filtered and the filtrate was concentrated.
The resulting residue was purified by column chroma-
tography (7:1, petroleum ether–EtOAc) to give 9
(2.33 g, 78% for two steps) as a syrup: [a]D ꢀ15.6 (c
3.4. 6-O-Acetyl-3-O-allyl-2,4-di-O-benzoyl-a-D-gluco-
pyranosyl trichloroacetimidate (8)
A
mixture of 4 (12 g, 54.5 mmol) and chloro-
triphenylmethane (18.2 g, 65.6 mmol) in pyridine
(80 mL) was stirred vigorously for 48 h at rt, at the
end of which time TLC (3:1, petroleum ether–EtOAc)
indicated that the reaction was complete. The mixture
was cooled to 0 ꢁC, then BzCl (21 mL, 180 mmol) was
added dropwise over 30 min before the reaction was
warmed to rt. The mixture was stirred for 24 h and then
water (200 mL) was added and stirring was continued
for 30 min. The mixture was extracted with CH2Cl2
and the combined extracts were washed with 1 N HCl
and saturated aq NaHCO3, dried (Na2SO4), and con-
centrated to a syrup that was purified by column chro-
matography (7:1, petroleum ether–EtOAc) to give 5
(29.8 g, 71%) as a white solid. A mixture of compound
5 (15.5 g, 20 mmol) and 150 mL 0.1% AcCl solution of
2:1 CH2Cl2–CH3OH (v/v) was left at rt about 1 h, at
the end of which time TLC (3:1, petroleum ether–
EtOAc) indicated the reaction was complete. The solu-
tion was neutralized with Et3N and concentrated. The
resultant residue was purified by column chromatogra-
phy (3:1, petroleum ether–EtOAc) to give crystalline 6
(9.6 g, 90%). Compound 6 (8 g, 15 mmol) was acetylated
with Ac2O (20 mL) in pyridine (20 mL) for 5 h at rt. The
mixture was concentrated and the residue was dissolved
in a solution of 3:2 THF–CH3OH (150 mL) and satu-
rated dry NH3. 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
solution was then concentrated and the residue was dis-
solved in CH2Cl2 (40 mL). To the solution were added
K2CO3 (2 g), CCl3CN (2 mL) and the mixture was stir-
red at rt for 12 h. The mixture was filtered, the filtrate
concentrated, and the residue was purified by column
chromatography (3:1, petroleum ether–EtOAc) to afford
8 (5.7 g, 77% for three steps) as a white solid: [a]D +62.4
1
1.0, CHCl3); H NMR (CDCl3, 400 MHz): d 8.09–7.45
(m, 10H, BzH), 5.32 (t, 1H, J = 9.5 Hz, H-4), 5.19 (dd,
1H, J = 7.8, 9.3 Hz, H-2), 4.70 (d, 1H, J = 7.8 Hz, H-
1), 4.30 (d, 2H, J = 4.0 Hz), 4.10 (t, 1H, J = 9.3 Hz,
H-3), 3.96–3.88 (m, 2H), 3.54 (m, 1H), 2.06 (s, 3H,
COCH3), 1.30–1.11 (m, 20H, CH2C10H20CH3), 0.91 (t,
3H, J = 6.9 Hz, CH2CH3). Anal. Calcd for C34H46O9:
C, 68.21; H, 7.74. Found: C, 68.12; H, 7.85.
3.6. Lauryl 2,3,4,6-tetra-O-benzoyl-b-D-galactopyran-
osyl-(1!4)-2,3,6-tri-O-benzoyl-b-D-glucopyranosyl-
(1!3)-2,4-di-O-benzoyl-b-D-glucopyranoside (10)
Trimethylsilyl trifluoromethanesulfonate (30 lL) was
added to a stirred solution of 3 (4.0 g, 3.3 mmol) and
9 (1.8 g, 3.0 mmol) in dry CH2Cl2 (40 mL) at rt. The
reaction mixture was stirred for 2 h, at the end of which
time TLC (3:1, petroleum ether–EtOAc) indicated that
the reaction was complete. The mixture was neutralized
with Et3N and concentrated. The resulting residue was
dissolved in a solution of CH3OH (50 mL) and CH2Cl2
(10 mL). To the solution was added AcCl (0.3 mL) and
the mixture was stirred for 7 h at rt, at the end of the
which time TLC (2:1, petroleum ether–EtOAc) indicated
that the reaction was complete. The mixture was neu-
tralized with Et3N, concentrated, and the residue was
purified by column chromatography (3:1, petroleum
ether–EtOAc) to give 10 (4.1 g, 85% for two steps) as
1
(c 1.0, CHCl3); H NMR (400 MHz, CDCl3): d 8.60 (s,
1
1H, NH), 8.09–7.42 (m, 10H, BzH), 6.69 (d, 1H,
J = 3.6 Hz, H-1), 5.62 (m, 1H, CH2–CH–CH2), 5.51 (t,
1H, J = 10.0 Hz, H-4), 5.42 (dd, 1H, J = 3.6, 10.0 Hz,
H-2), 5.06 (dd, 1H, J = 1.6, 17.6 Hz, CH2–CH–CH2),
4.96 (dd, 1H, J = 1.6, 10.4 Hz, CH2–CH–CH2), 4.35–
4.21 (m, 4H, H-3, H-5, CH2–CH–CH2), 4.19–4.07 (m,
2H, H-6), 2.06 (s, 3H, COCH3). Anal. Calcd for
C27H26Cl3NO9: C, 52.74; H, 4.26. Found: C, 52.24; H,
4.64.
a white amorphous solid: [a]D +9.2 (c 1.0, CHCl3); H
NMR (CDCl3, 400 MHz): d 7.99–7.09 (m, 45H, BzH),
5.62–5.58 (m, 2H), 5.54 (t, 1H, J = 9.7 Hz), 5.38 (dd,
1H, J = 7.8, 9.5 Hz), 5.32 (t, 1H, J = 9.3 Hz), 5.26 (t,
1H, J = 9.0 Hz), 5.18 (dd, 1H, J = 3.3, 10.2 Hz), 4.90
(d, 1H, J = 7.8 Hz), 4.55 (d, 2H, J = 7.8 Hz), 4.36 (t,
1H, J = 9.0 Hz), 4.20–4.19 (m, 2H), 4.03 (t, 1H,
J = 9.5 Hz), 3.84–3.73 (m, 2H), 3.67–3.58 (m, 5H),
3.48–3.33 (m, 2H), 1.31–1.00 (m, 20H, CH2C10H20CH3),

2350
X. Mei et al. / Carbohydrate Research 340 (2005) 2345–2351
0.91 (t, 3H, J = 6.8 Hz, CH2CH3); 13C NMR (100 MHz,
CDCl3): d 165.45, 165.35, 165.25, 165.20, 165.13, 165.00,
164.84, 164.58, 164.18 (9C, C(@O)Ph), 133.10–127.93
(Ph), 100.91, 100.78, 100.51 (3C, C-1A, 1B, 1C). Anal.
Calcd for C93H92O25: C, 69.39; H, 5.76. Found: C,
69.26; H, 5.87.
ether–EtOAc) to give 16 (5.3 g, 87%) as a white amor-
phous solid: [a]D ꢀ12.0 (c 1.0, CHCl3); 1H NMR
(CDCl3, 400 MHz): d 7.93–7.01 (m, 65H, BzH), 5.84
(t, 1H, J = 9.6 Hz), 5.69–5.64 (m, 2H), 5.58–5.46 (m,
3H), 5.34 (m, 1H), 5.23–5.08 (m, 5H), 5.02–4.98 (m,
2H), 4.85–4.71 (m, 5H), 4.64–4.59 (m, 2H), 4.50–4.43
(m, 3H), 4.35–4.32 (m, 2H), 4.26–4.22 (m, 3H), 4.15–
4.12 (m, 2H), 3.99–3.96 (m, 5H), 3.93–3.66 (m, 7H),
3.64–3.58 (m, 5H), 3.55–3.44 (m, 6H), 3.31 (m, 1H),
3.23 (m, 1H), 2.07–2.02 (m, 15H), 1.90 (s, 3H), 1.84 (s,
3H), 1.82 (s, 3H), 1.30–0.95 (m, 20H), 0.89 (t, 3H,
J = 6.9 Hz); 13C NMR (CDCl3, 100 MHz): d 170.44,
170.39, 169.46, 169.11, 169.04, 168.94, 168.75, 168.47
(8C, C(@O)CH3), 165.90, 165.67, 165.33, 165.17,
165.11, 165.07, 165.03, 165.00, 164.82, 164.75, 164.59,
164.44, 164.18 (13C, C(@O)Ph), 133.99–127.93 (Ph,
CH2CH@CH2), 116.81 (CH2CH@CH2), 101.07,
100.85, 100.73, 100.53, 100.45, 100.43, 100.10 (7C,
C-1A, 1B, 1C, 1D, 1E, 1F, 1G), 45.65 (1C, CH2CH@CH2).
Anal. Calcd for C164H168O57: C, 64.56; H, 5.55. Found:
C, 64.49; H, 5.74.
3.7. Isopropyl 2,4,6-tri-O-acetyl-3-O-allyl-b-D-gluco-
pyranosyl-(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-gluco-
pyranosyl-(1!6)]-2,4-di-O-acetyl-b-D-glucopyranosyl-
(1!3)-2,4,6-tri-O-acetyl-1-thio-b-D-glucopyranoside (15)
Trimethylsilyl trifluoromethanesulfonate (30 lL) was
added to a stirred solution of 11 (6.55 g, 5.0 mmol)
and 12 (1.65 g, 5.0 mmol) in dry CH2Cl2 (40 mL) at rt.
The reaction mixture was stirred for 2 h, at the end of
which time TLC (3:1, petroleum ether–EtOAc) indicated
that the reaction was complete. The mixture was neu-
tralized with Et3N and concentrated. The resultant resi-
due was added to 90% HOAc/H2O (40 mL), the mixture
was heated at reflux for 2 h. The mixture was concen-
trated and the residue was acetylated with Ac2O
(20 mL) in pyridine (20 mL) for 5 h at rt. The solvent
was evaporated and the resulting residue was purified
by column chromatography (2:1, petroleum ether–
EtOAc) to afford 15 (4.9 g, 92% for three steps) as a
white solid: [a]D ꢀ29.5 (c 1.0, CHCl3); 1H NMR
(CDCl3, 400 MHz): d 8.01–7.26 (m, 20H, BzH), 5.87–
5.68 (m, 3H), 5.58–5.46 (m, 3H), 5.62 (m, 2H), 5.18–
5.10 (m, 2H), 5.03–4.94 (m, 3H), 4.85 (m, 1H), 4.66–
4.45 (m, 4H), 4.39–4.20 (m, 5H), 4.03–3.88 (m, 6H),
3.83–3.70 (m, 4H), 3.67–3.45 (m, 3H), 3.18 (m, 1H,
SCH(CH3)2), 2.11–1.91 (8s, 24H, COCH3), 1.33–1.25
(m, 6H, SCH(CH3)2); 13C NMR (CDCl3, 100 MHz): d
170.67, 170.60, 169.57, 169.30, 169.17, 168.96, 168.84,
168.60 (8C, C(@O)CH3), 166.05, 165.84, 165.16,
164.99 (4C, C(@O)Ph), 134.09–128.32 (Ph), 116.97
(CH2CH@CH2), 101.25, 100.48, 100.27 (3C, C-1B, 1C,
1D), 83.10 (C-1A), 35.36 (1C, SCH(CH3)2), 24.44, 24.16
(2C, SCH(CH3)2). Anal. Calcd for C74H84O32S: C,
58.57; H, 5.58. Found: C, 58.44; H, 5.71.
3.9. Lauryl b-D-glucopyranosyl-(1!3)-[b-D-glucopyran-
osyl-(1!6)]-b-D-glucopyranosyl-(1!3)-b-D-glucopyran-
osyl-(1!6)-[b-D-galactopyranosyl-(1!4)-b-D-gluco-
pyranosyl-(1!3)]-b-D-glucopyranoside (17)
Compound 16 (3.7 g, 1.2 mmol) was dissolved in a solu-
tion of CH3OH (40 mL) and CH2Cl2 (20 mL). To this
solution was added PdCl2 (40 mg) and the mixture was
stirred for 6 h at rt, at the end of the which time TLC
(1:1, petroleum ether–EtOAc) indicated that the reac-
tion was complete. The mixture was filtered, the filtrate
concentrated and the residue was then dissolved in a sat-
urated solution of NH3 in CH2Cl2 (30 mL) and CH3OH
(300 mL) at rt. After 96 h, the reaction mixture was con-
centrated, and the residue was washed four times with
CH2Cl2 to afford 17 (1.3 g, 82% for two steps) as a white
solid: [a]D ꢀ22.6 (c 1.0, H2O). 1H NMR (D2O,
400 MHz): d 4.84–4.62 (m, 4H), 4.38–4.31 (m, 3H),
4.07–3.77 (m, 9H), 3.60–3.34 (m, 18H), 3.29–3.05 (m,
17H), 1.47–1.12 (m, 20H, CH2C10H20CH3), 0.73 (t,
3H, J = 6.6 Hz, CH2CH3); 13C NMR (CDCl3,
100 MHz): d 103.14, 102.08, 102.44, 102.04, 101.85,
101.24, 101.09 (C-1A–G). Anal. Calcd for C54H96O36:
C, 49.09; H, 7.32. Found: C, 48.57; H, 7.69. ESIMS
for C54H96O36 (1321.33): 1320.32 [Mꢀ1]+.
3.8. Lauryl 2,4,6-tri-O-acetyl-3-allyl-b-D-glucopyranosyl-
(1!3)-[2,3,4,6-tetra-O-benzoyl-b-D-glucopyranosyl-
(1!6)]-2,4-di-O-acetyl-b-D-glucopyranosyl-(1!3)-2,4,6-
tri-O-acetyl-b-D-glucopyranosyl-(1!6)-[2,3,4,6-tetra-O-
benzoyl-b-D-galactopyranosyl-(1!4)-2,3,6-tri-O-benzoyl-
b-D-glucopyranosyl-(1!3)]-2,4-di-O-benzoyl-b-D-gluco-
pyranoside (16)
Acknowledgements
To a stirred solution of 10 (3.2 g, 2.0 mmol) and 15
(3.3 g, 2.2 mmol) in dry CH2Cl2 (50 mL) were added
N-iodosuccinimide (0.5 g, 2.2 mmol) and TMSOTf
(15 lL) at rt. After 2 h, Et3N was added and the mixture
was filtered and the filtrate concentrated. The residue
was purified by column chromatography (2:1, petroleum
This work was supported by The Natural Science Fund
from the Educational Committee of Sichuan Province
(2003407) and The Natural Science Fund from Chongq-
ing University of Post and Telecommunications
(A2004–45).

X. Mei et al. / Carbohydrate Research 340 (2005) 2345–2351
2351
16. Ning, J.; Yi, Y.; Kong, F. Tetrahedron Lett. 2002, 43,
5545–5549.
References
17. (a) Xing, Y.; Ning, J. Tetrahedron: Asymmetry 2003, 14,
1275–1283; (b) Ning, J.; Heng, L.; Kong, F. Tetrahedron
Lett. 2002, 43, 673–675.
18. Ning, J.; Yi, Y.; Yao, Z. Synlett 2003, 14, 2208–2212.
19. (a) Ning, J.; Wang, H.; Yi, Y. Tetrahedron Lett. 2002, 43,
7349–7352; (b) Wang, H.; Ning, J. J. Org. Chem. 2003, 68,
2521–2524.
20. (a) Schmidt, R. R.; Michel, J. Angew. Chem., Int. Ed. Engl.
1980, 19, 731–732; (b) Schmidt, R. R. Angew. Chem., Int.
Ed. Engl. 1986, 25, 212–235; (c) Rademann, J.; Geyer, A.;
Schmidt, R. R. Angew. Chem., Int. Ed. Engl. 1998, 37,
1241–1245.
21. Liang, R. Science 1996, 274, 1520–1522.
22. (a) Martin, T. J.; Schmidt, R. R. Tetrahedron Lett. 1992,
33, 6123–6126; (b) Kondo, H.; Ichikawa, Y.; Wong, C. H.
J. Am. Chem. Soc. 1992, 114, 8748–8750.
23. Hashimoto, S.; Honda, T.; Ikegami, S. J. Chem. Soc.,
Chem. Commun. 1989, 11, 685–687.
24. Ferrier, R. J.; Hay, R. W.; Vethaviyasar, N. Carbohydr.
Res. 1973, 27, 55–61.
25. Fraser-Reid, B.; Merritt, R.; Handlon, A. L.; Andrews, C.
W. Pure Appl. Chem. 1993, 65, 779–784.
26. Schmidt, R. R.; Kinzy, W. Adv. Carbohydr. Chem.
Biochem. 1994, 50, 21–125.
1. (a) Vicker, M. G. J. Cell. Sci. 1976, 21, 161–173; (b) Asao,
M. I.; Oppenheimer, S. B. Exp. Cell Res. 1979, 120, 101–
110; (c) Costello, M.; Fiedel, B. A.; Gewurz, H. Nature
1979, 281, 677–678.
2. Suzuki, T.; Tsuzuki, A.; Ohno, N.; Ohshima, Y.; Adachi,
Y.; Yadomae, T. Biol. Pharm. Bull. 2002, 25, 140–144.
3. Raz, A.; Lotan, R. Cancer Res. 1981, 41, 3642–3647.
4. Lotan, R.; Raz, A. Cancer Res. 1983, 43, 2088–2093.
5. Kikumoto, S.; Miyajima, T.; Yoshizumi, S.; Fujimoto, S.;
Kimura, K. J. Agric. Chem. Soc. Jpn. 1970, 44, 337–342.
6. Kikumoto, S.; Miyajima, T.; Kimura, K.; Okubo, S.;
Komatsu, N. J. Agric. Chem. Soc. Jpn. 1971, 45, 162–
168.
7. Komatsu, N.; Okubo, S.; Kikumoto, S.; Kimura, K.;
Saito, G.; Sakai, S. Gann 1969, 60, 137–144.
8. Komatsu, N.; Nagumo, N.; Okubo, S.; Koike, K. Jpn. J.
Antibiot. 1973, 26, 277–283.
9. Koike, K. Jpn. J. Antibiot. 1976, 29, 1098–1105.
10. Tabata, K.; Ito, W.; Kojima, T.; Kawabata, S.; Misaki, A.
Carbohydr. Res. 1981, 89, 121–135.
11. (a) Sakagami, H.; Kim, F.; Konno, K. Anticancer Res.
1990, 10, 697–702; (b) Sakagami, H.; Aoki, T.; Simpson,
A.; Tanuma, S. I. Anticancer Res. 1991, 11, 993–1000.
12. Ohno, N.; Miura, N. N.; Chiba, N.; Adachi, Y.; Yado-
mae, T. Biol. Pharm. Bull. 1995, 18, 1242–1247.
13. Usui, S.; Tomono, Y.; Sakai, M.; Kiho, T.; Ukai, S. Biol.
Pharm. Bull. 1995, 18, 1630–1636.
27. Ning, J.; Zhang, W.; Yi, Y.; Yang, G.; Wu, Z.; Yi, Z.;
Kong, F. Bioorg. Med. Chem. 2003, 11, 2193–2203.
28. He, H.; Yang, F.; Du, Y. Carbohydr. Res. 2002, 337, 1673–
1678.
`
´
14. Jamois, F.; Ferrieres, V.; Guegan, J. P.; Yvin, J. C.;
Plusquellec, D.; Vetvicka, V. Glycobiology 2005, 15, 393–
407.
´
29. Bairamova, N. E.; Bakinovskii, L. V.; Kochetkov, N. K.
Russ. Chem. Bull. 1985, 34, 1024–1030.
30. Li, A.; Kong, F. Carbohydr. Res. 2004, 339, 2499–2506.
15. Sears, P.; Wong, C. H. Science 2001, 291, 2344–2350.