Synthesis of complex-type glycans derived from parasitic helminths

Article abstract of DOI:10.1016/j.carres.2006.12.022

Chemical syntheses of complex-type glycans derived from the eggs of parasitic helminths, Schistosoma mansoni and Schistosoma japonicum were achieved. In addition, their analogs, which lack xylose and/or fucose residue(s), are described. These branched sug

Full text of DOI:10.1016/j.carres.2006.12.022

Carbohydrate Research 342 (2007) 675–695
Synthesis of complex-type glycans derived from parasitic helminths
Jun Nakano,a,b Akihiro Ishiwata,a,c Hiromichi Ohtab and Yukishige Itoa,c,
*
aRIKEN (The Institute of Physical and Chemical Research), Wako, Saitama 351-0198, Japan
bDepartment of Biosciences and Informatics, Keio University, Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan
cCREST, JST, Kawaguchi, Saitama 332-0012, Japan
Received 14 November 2006; received in revised form 22 December 2006; accepted 22 December 2006
Available online 4 January 2007
Abstract—Chemical syntheses of complex-type glycans derived from the eggs of parasitic helminths, Schistosoma mansoni and Schis-
tosoma japonicum were achieved. In addition, their analogs, which lack xylose and/or fucose residue(s), are described. These
branched sugar chains were synthesized regio- and stereoselectively by using b-mannosylation, desilylation under high-pressure
and glycosylation in frozen solvent as key transformations.
ꢀ 2007 Elsevier Ltd. All rights reserved.
Keywords: Parasitic helminths; Glycoprotein; Complex-type glycans
1. Introduction
to the synthesis of complex-type N-glycans 1 and 2,
which were found in the eggs of parasitic helminths,
Schistosoma mansoni and Schistosoma japonicum (Chart
1).8 They share structural elements with higher eukary-
otes. Namely, they are linked to the side chain of Asn
through GlcNAc and carry the common pentasaccha-
ride Man3GlcNAc2. However, they have relatively short
chain length, lacking outer galactose and sialic acid
residues. They are instead decorated by D-xylose and
L-fucose residues, which are linked to mannose
(Xylb1!2Man) and innermost N-acetylglucosamine
(Fuca1!3GlcNAc), respectively. Interestingly, plant
derived N-glycans also have these residues. It has been
shown that these structures are antigenic to human,9
and contribute to IgE binding to plant allergens.10
Helminth N-glycans have an additional Fuc, which is
a-(1!6) linked to the innermost GlcNAc, as is often
observed in mammalian complex-type glycans.
A majority of proteins produced by eukaryotes are
glycoproteins, which carry asparagine (Asn)-linked
(N-linked) oligosaccharides (N-glycans).1 Although these
glycans are structurally diverse, they share a common
feature, carrying the core pentasaccharide that consists
of three mannose and two N-acetylglucosamine residues
(Man3GlcNAc2). Eukaryotic N-glycans are introduced
in the endoplasmic reticulum (ER) as a tetradecasaccha-
ride (Glc3Man9GlcNAc2). They are then processed by
various glycosidases and glycosyltransferases to gener-
ate highly diverse glycans.2 N-Glycans play important
roles in numerous biological events, such as develop-
ment, signal transduction, cell–cell recognition, malig-
nant transformation, and immune response.3 They are
also functional in modulating protein folding, transport,
and degradation.4
Recently, novel N-glycans have been identified from
bacteria,5 plants,6 and parasites.7 They are attracting
attention, in terms of relationships with allergy, infec-
tion, and pathogenicity. Our interest has been directed
Schistosomes chronically infect more than 200 million
people in developing countries.11 Infection with S. man-
soni induces TH2 type immune response,12 which was
ascribed to adjuvant activities of carbohydrates.13 Curi-
ously, individuals infected with the parasite acquire
resistance to allergy.14 The so-called ‘IgE blocking
hypothesis’ implies that the polyclonal IgE antibody
produced after parasite infection saturates the IgE
*
Corresponding author. Tel.: +81 48 467 9430; fax: +81 48 462
4680; e-mail: yukito@riken.jp
0008-6215/$ - see front matter ꢀ 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2006.12.022

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J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
Chart 1. Typical structures of animal (A), plant (B), and helminth (C) derived N-glycans. (D) Oligosaccharides synthesized in this study.
receptors on mast cells and blocks the binding of specific
IgE antibody. On the other hand, the rapid increase of
allergic diseases in urban area is explained by the
‘hygiene hypothesis’. It advocates that the highly hygienic
environment caused drastically reduced infection, which
promotes the outbreak of the allergy.15 More recently, a
new theory has emerged, which advocates the role of
IL-10 in anti-inflammatory network for inhibiting the
allergy. Long-term parasite infection upregulates the
production of this cytokine in the presence of regulatory
T cells. However, a precise understanding of the roles of
glycoprotein glycans in these phenomena has been diffi-
cult to identify, because of the limited access to these
molecules.
to be challenging. Described herein is the full detail of
our synthetic studies toward mono- (2) and difucosyl-
ated (1) xylosylated glycans,16 as well as a series of their
analogs systematically lacking Fuc and/or Xyl residues
(3–8). It is our perspective that a comparison of their
activities may provide information as for the contribu-
tion of each sugar residue on the immunostimulant
activity of helminth/plant glycans.
2. Results and discussion
2.1. General synthetic plan
In addition to their biomedical significance, highly
complex branching of 1 and 2 attracted our attention.
In particular, the construction of triply branched substi-
tution patterns on GlcNAca and Manc was anticipated
In order to approach 1–8 in a systematic manner, we set-
tled on the use of xylosylated (9) and non-xylosylated
(12) donors, (1!6)-fucosylated (10), and non-fucosyl-
ated (13) glucosamine components, and fucosyl donor

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
677
Chart 2. Design of synthetic blocks.
1117 (Chart 2). We expected that the combination of
these fragments [(9 or 12) · (10 or 13) · (+ or ꢀ11)]
would provide 1–8, with a uniform set of reactions.
and 20b) were prepared (Scheme 1). To begin with, com-
pound 1418 was subjected to the intramolecular aglycon
delivery process (IAD) using 15 as a donor19 to give 17
as a pure b-isomer, through intermediacy of hemiacetal
16. This reaction was particularly suitable for our pur-
pose, because it simultaneously liberated the 2-OH of
b-mannose. Thus, product 17 was immediately subjected
to the next glycosylation with trichloroacetimidate20
2.2. Synthesis of hexa- and pentasaccharide donors
As precursors of hexa- and pentasaccharide donors
(9 and 12), b-mannoside containing fragments (20a

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J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
˚
Scheme 1. Reagents and conditions: (1) 15 (1.2 equiv), DDQ (1.25 equiv), MS 4 A, CH2Cl2, rt, 1.5 h; (2) MeOTf (3.5 equiv), DTBMP (4 equiv), MS
˚
4 A, (ClCH2)2, 45 ꢁC, 24 h, 82% (two steps); (3) 18 (3 equiv), TMSOTf (2 equiv), CH2Cl2, ꢀ40 ꢁC, 2 h, 67% for 19a; (4) Ac2O, Py, DMAP (0.1 equiv),
rt, 12 h, 97%; (5) HFÆPy, DMF, 1 GPa, 12 h, 88% for 20a, 89% for 20b.
1821 as a xylosyl donor to afford 19a. This reaction
required somewhat forcing conditions (3 equiv of 18,
2 equiv of TMSOTf), reflecting the steric hindrance of
17; the reacting OH was axially orientated and had unfa-
vorable gauche interactions with two bulky groups
(OTBDPS and heavily protected glucosamine).
to give hexa- 27a and pentasaccharide 27b, without
complexity.
In order to convert them to hexa/pentasaccharide
donors, our initial plan was to undertake the selective
deprotection of the anomeric benzyl group. Thus, 27a
was acetylated and treated under modified Bieg condi-
tions27 using Pd–Al2O3 as a catalyst and cyclohexene
as a hydrogen source. In fact, this reaction indeed gave
hemiacetal 28a in 60% yield. However, its isolation was
difficult by the contamination of other debenzylated
products. We then settled on a three-step procedure,
which involved complete debenzylation, peracetylation
and selective deacetylation.
Complete debenzylation of 27a was far less straight-
forward than expected. Namely, hydrogenolysis under
standard conditions [H2, Pd(OH)2, EtOH–EtOAc–
water] and subsequent acetylation gave rise to the for-
mation of side products having one of their phthaloyl
groups saturated. After extensive screening, it was found
that this complication could be avoided by employing
the hydrogen transfer protocol28 using Pd(OH)2 in
2:1:1 cyclohexane–EtOH–AcOH.ꢀ The debenzylated
product was isolated as peracetate 28b, which was con-
verted to trichloroacetimidate 9 in a standard fashion.
In order to liberate the hydroxy group for further gly-
cosylation, 19a was subjected to the deprotection of the
TBDPS group. Treatment with tetra-n-butylammonium
fluoride (TBAF) and acetic acid (1:1) in DMF gave the
desilylated product 20a in 75% yield. However, in this
case, the product isolation was rather cumbersome, pre-
sumably due to the occurrence of acetyl migration. This
transformation was achieved more cleanly under high-
pressure conditions22 using HF-pyridine to provide 20a
in 88% yield. In comparison, when the same reaction
was conducted under atmospheric pressure, otherwise
identical conditions, no product formation was observed.
In the same fashion, desilylation of compound 19b,
which was obtained by acetylation of 17, provided 20b.
Preparation of the GlcNAc1!2Man component 23
was conducted by the reaction of 2123 and chloride
2224 under standard conditions (Scheme 2). Disaccha-
ride 23 was then coupled with 20a and 20b, through acti-
vation with MeOTf,25 to give 24a and 24b, respectively.
Both were converted to diols 25a and 25b after acidic
removal of the cyclohexylidene group. Glycosylation
with mannosyl chloride 2626 proceeded regioselectively
ꢀ The proportion of the solvent was critical. A smaller proportion of
AcOH resulted in incomplete deprotection.

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
679
˚
Scheme 2. Reagents and conditions: (5) 22 (1.27 equiv), AgOTf (2.5 equiv), MS 4 A, (ClCH2)2–toluene (2:1), ꢀ40 ꢁC, 1 h, 63%; (1) 23 (1.5 equiv),
˚
MeOTf (4.5 equiv), DTBMP (4.5 equiv), MS 4 A, toluene, 50 ꢁC, 12 h, 94% for 24a; 9 h, 79% for 24b; (2) TsOHÆH2O (2.5 equiv), CH3CN, rt, 9 h,
˚
91% for 25a; 12 h, 86% for 25b; (3) 26 (1.2 equiv), AgOTf (2.4 equiv), MS 4 A, (ClCH2)2–toluene (2 :1), ꢀ30 ꢁC to rt, 2.5 h, 77% for 27a; 6 h, 89% for
27b; (4) Pd(OH)2, cyclohexene–EtOH–AcOH (2:1:1), reflux, 60 h; (5) Ac2O, Py, rt, 6 h, 93% for 28b (two steps); 7 h, 94% for 28c (two steps); (6)
N2H4ÆAcOH (1.3 equiv), DMF, rt, 2 h; (7) DBU (0.95 equiv), Cl3CCN, rt, 12 h, 75% for 9, (two steps); 74% for 12 (two steps).
In the same manner, the non-xylosylated pentasaccha-
ride 27b was converted to 12 via 28c.
ated disaccharide 30 as an alternative acceptor.
However, its reaction with 9 gave the coupled product
only in low (<10%) yield.
2.3. Synthesis of xylosylated glycans
We selected 6-O-fucosylated disaccharide 10 as an
acceptor, hoping that it had less steric hindrance than
29 or 30, because of the conformational flexibility of
the C-6 position. The synthesis of 10 was conducted as
shown in Scheme 3. Thus, compound 3118 was con-
verted to its p-methoxybenzyl (PMB) ether 32 using
With designed donors 9 and 12 in place, we initially
explored the possibility to combine them with difucosyl-
ated trisaccharide, to complete the synthesis of 1 and 6
in a most convergent manner. To explore this possibil-
ity, the reaction of 9 with a potential acceptor 29 was
conducted (Scheme 3). However, several attempts to
achieve this coupling resulted in complete failure, sug-
gesting the severe congestion of the hydroxy group,
which was sandwiched by two sugar residues. To allevi-
ate the steric hindrance, we then prepared 3-O-fucosyl-
29
PMB trichloroacetimidate and La(OTf)2 and then to
diol 33. The latter was regioselectively glycosylated with
11 to give 10. To our delight, the coupling of 9 with
10 (1.5 equiv) proceeded smoothly under standard
trichloroacetimidate conditions20 to give octasaccharide
34 in a satisfactory yield (Scheme 4).

680
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
Scheme 3. Reagents and conditions: (1) 30 (3 equiv), La(OTf)3 (0.12 equiv), CH2Cl2, rt, 20 h, 79%; (2) TsOHÆH2O (3.5 equiv), CH3CN–MeOH (1:1),
˚
rt, 11 h, 78%; (3) 11 (1.2 equiv), MeOTf (3.6 equiv), DTBMP (3.6 equiv), MS 4 A, CPME, rt, 22 h, 66%; (4) BH3ÆNMe3 (5 equiv), A1Cl3 (5 equiv),
THF, rt, 20 h, 66%.
In order to introduce the (1!3)-linked fucose, the
p-methoxybenzyl group of 34 had to be removed. How-
ever, under standard conditions (DDQ, H2O, CH2Cl2),
this reaction was rather sluggish, requiring 6.5 equiv of
DDQ for completion. Although the desired 35 was
obtained, the yield was only modest (56%) and concom-
itant formation of monodebenzylated product (18%)
hampered the facile isolation of 35. This difficulty was
alleviated by using manganese(III) acetate as a co-
oxidant.30 Namely, treatment of 34 with a small excess
of DDQ and 3.6 equiv of Mn(OAc)3Æ2H2O in CH2Cl2
gave 97% yield of 35. Conversion of 9 to 37 was con-
ducted in a similar manner; glycosylation with 13 was
followed by the removal of the PMB group to give 37
in 72% yield from 9.
we decided to apply these conditions to the coupling
between 35 and 11. Gratifyingly, when the coupling
was conducted in p-xylene at 4 ꢁC, nearly complete
conversion was achieved and nonasaccharide 36 was ob-
tained in 90% yield. On another hand, fucosylation of 37
with 11 proceeded smoothly under standard solution
conditions, possibly reflecting the reduced steric hin-
drance of 37 compared to 35, providing 38 in high yield.
With the successfully assembled nona-36, hepta-34,
38, and hexasaccharide 37 in hand, these compounds
were subjected to complete deprotection in a uniform
manner. Thus, sequential dephthaloylation, acetylation,
O-deacetylation, and debenzylation provided 1 (X1F2),
2 (X1F1-A), 3 (X1F0), and 4 (X1F1-B).
Introduction of the fucose residue to 35 was con-
ducted using 11, through activation with MeOTf. Since
this reaction was anticipated to be difficult, we were
encouraged to observe the formation of the desired
product 36 in ꢁ50% yield (estimated from MALDI-
TOFMS) by using ꢁ5 equiv of 11. However, the
separation of 36 from unreacted 35 was impossible in
a preparative scale and we were obliged to identify the
conditions that lead to complete consumption of 35.
Since we recently found that MeOTf promoted glycosyl-
ation using methylthio glycoside could be accelerated
under frozen conditions in p-xylene (mp 12–13 ꢁC),31
2.4. Synthesis of non-xylosylated glycans
The synthesis of a series of non-xylosylated glycans 5–8
was conducted, essentially as described for 1–4, except
that pentasaccharide 12 was employed as a common
donor (Scheme 5). Coupling with disaccharide 10 gave
39, which was converted to 40 and glycosylated with
11 under frozen conditions to give 41. On the other
hand, coupling of 12 and 13 (2 equiv) was followed by
PMB deprotection to give 42 in 81% yield. Further gly-
cosylation with 11 gave 43. Compounds 39, 41, 42, and

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
681
Scheme 4. Reagents and conditions: (1) 10 (1.5 equiv), TMSOTf (0.2 equiv), CH2Cl2, ꢀ78 to ꢀ40 ꢁC, 4.5 h, 74%; (2) DDQ (1.2 equiv),
˚
Mn(OAc)3Æ2H2O (3.6 equiv), rt, 24 h, 97%; (3) 11 (3 equiv), MeOTf (7.5 equiv), DTBMP (4.5 equiv), MS 4 A, p-xylene, 4 ꢁC, 90%; (4) 13 (2 equiv),
TMSOTf (0.2 equiv), CH2Cl2, ꢀ78 to ꢀ50 ꢁC, 1.5 h; (5) DDQ (0.96 equiv), Mn(OAc)3Æ2H2O (2.9 equiv), CH2Cl2, rt, 12 h, 72% (two steps); (6) 11
˚
(2 equiv), MeOTf (6 equiv), DTBMP (5 equiv), MS 4 A, CPME, rt, 23 h, 85%; (7) (H2NCH2)2, n-BuOH, 85 ꢁC, 12 h; (8) Ac2O, Py, rt, 6 h; (9)
MeONa, MeOH, rt, 12 h; (10) Pd(OH)2, aq MeOH, rt, 12 h, 87% for 1 (four steps), 73% for 2 (four steps), 88% for 3 (four steps), 68% for 4 (four
steps).
43 thus obtained were deprotected to give 5 (X0F1-A), 6
(X0F2), 7 (X0F0), and 8 (X0F1-B).
JMS-T700LCK equipment with CF3CO2Na as an inter-
nal standard.
In conclusion, the systematic synthesis of complex-
type N-glycans 1 and 2 found in the eggs of parasites,
S. mansoni and S. japonicum, as well as their analogs
lacking fucose and/or xylose residues was accomplished.
These compounds are expected to be valuable in order
to reveal the structure activity relationship of plant-
and helminth-derived oligosaccharides.
3.2. Benzyl (3-O-tert-butyldiphenylsilyl-4,6-O-cyclohexyl-
idene-b-D-mannopyranosyl)-(1!4)-3,6-di-O-benzyl-2-
deoxy-2-phthalimido-b-D-glucopyranoside (17)
To a mixture of 15 (48.5 mg, 83.7 lmol) and 14
˚
(57.0 mg, 87.8 lmol) and preactivated MS 4 A (0.5 g)
in dry CH2Cl2 (2.5 mL) was added DDQ (23.7 mg,
114 lmol) at 0 ꢁC, and the mixture was stirred for 3 h,
during which time the temperature was raised to room
temperature. The reaction was quenched with aq ascor-
bic acid (0.7%)–citric acid (1.3%)–NaOH (0.9%) and fil-
tered through Celite, which was rinsed with EtOAc. The
filtrate was separated and the organic layer was washed
with satd aq NaHCO3 and brine, successively, and dried
over Na2SO4. After concentration under diminished
pressure, the mixed acetal 16 was used for the next reac-
tion without further purification. To a mixture of mixed
acetal 16 and DTBMP (68.8 mg, 335 lmol), which were
coevaporated with toluene, in dry 1,2-dichloroethane
3. Experimental
3.1. General
1H and 13C NMR spectra were measured on a JEOL
EX-400 spectrometer in CDCl3 and were referenced to
Me4Si unless otherwise stated. Silica gel column chro-
matography was performed using Silica Gel-60 (E.
Merck). MALDI-TOFMS spectra were recorded in the
positive ion mode on an AXIMA CFR (Shimadzu/
KRATOS) equipped with a nitrogen laser with an emis-
sion wavelength of 337 nm. High-resolution ESI-TOF
mass spectra were obtained with a JEOL AccuTOF
˚
(8.0 mL) were added MS 4 A (0.9 g) and MeOTf
(32.1 lL, 284 lmol) at room temperature. The mixture
was stirred at 45 ꢁC for 15 h and cooled down to room

682
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
Scheme 5. Reagents and conditions: (1) 10 (1.5 equiv), TMSOTf (0.2 equiv), CH2Cl2, ꢀ78 to ꢀ40 ꢁC, 4.5 h, 70%; (2) DDQ (1.3 equiv),
˚
Mn(OAc)3Æ2H2O (3.9 equiv), rt, 13 h, 92%; (3) 11 (3 equiv), MeOTf (7.5 equiv), DTBMP (4.5 equiv), MS 4 A, p-xylene, 4 ꢁC, frozen condition, 84 h,
80%; (4) 13 (2 equiv), TMSOTf (0.2 equiv), CH2Cl2, ꢀ78 to ꢀ50 ꢁC, 2 h; (5) DDQ (1.05 equiv), Mn(OAc)3Æ2H2O (3.1 equiv), CH2Cl2, rt, 12 h, 81%
˚
(two steps); (6) 11 (2 equiv), MeOTf (5.5 equiv), DTBMP (5 equiv), MS 4 A, CPME, rt, 20.5 h, 92%; (7) (H2NCH2)2, n-BuOH, 85 ꢁC, 12 h; (8) Ac2O,
Py, rt, 6 h; (9) MeONa, MeOH, rt, 12 h; (10) Pd(OH)2, aq MeOH, rt, 12 h, 75% for 5 (four steps), 74% for 6 (four steps), 74% for 7 (four steps), 69%
for 8 (four steps).
temperature. The reaction was quenched with Et3N
(1.0 mL), eluted with EtOAc, and filtered through Cel-
ite, which was rinsed with EtOAc. The filtrate was
washed with satd aq NaHCO3 and brine, successively,
and dried over Na2SO4. After concentration under
diminished pressure, the residue was purified by
preparative TLC (2:1 hexane–EtOAc) to give 72.4 mg
(82% in two steps) of compound 17; [a]D +17.6 (c 1.0,
3.3. Benzyl (2,3,4-tri-O-acetyl-b-D-xylopyranosyl)-
(1!2)-(3-O-tert-butyldiphenylsilyl-4,6-O-cyclohexyl-
idene-b-D-mannopyranosyl)-(1!4)-3,6-di-O-benzyl-2-
deoxy-2-phthalimido-b-D-glucopyranoside (19a)
˚
A mixture of preactivated molecular sieves 4 A (120 mg)
and compounds 18 (63.1 mg, 150 lmol) and 17
(52.8 mg, 49.8 lmol) in CH2Cl2 (2.5 mL) was stirred at
ꢀ40 ꢁC for 30 min. TMSOTf (18 lL, 0.10 mmol) was
added and stirred at the same temperature for 2 h. The
reaction was quenched with Et3N (30 lL), stirred at
ꢀ40 ꢁC for 5 min and filtered through Celite. The filtrate
was diluted with EtOAc, washed with brine, dried over
MgSO4, and evaporated under diminished pressure.
The residue was purified by preparative TLC (3:2 hex-
ane–EtOAc) to afford 44.0 mg (67%) of compound 19a
and 6.2 mg (12%) of recovered 17. Compound 19a:
[a]D ꢀ34.4 (c 1.0, CHCl3); 1H NMR (400 MHz, CDCl3):
d 7.80–6.74 (m, 29H, Ar), 5.11–5.03 (m, 3H, H-1GlcN, H-
1,3Xyl), 4.96 (dd, J 5.1, 7.1 Hz, 1H, H-2Xyl), 4.80–4.69
(m, 3H, H-4Xyl, PhCH2), 4.52 (d, J 12.2 Hz, 1H,
PhCHH), 4.43 (d, J 12.5 Hz, 1H, PhHH), 4.35 (br s,
1H, H-1Man), 4.31 (d, J 12.2 Hz, 1H, PhHH), 4.20–
4.11 (m, 4H, H-2,3GlcN, H-5aXyl, PhCHH), 3.93 (t, J
1
CHCl3); H NMR (400 MHz, CDCl3): d 7.87–6.82 (m,
29H, Ar), 5.10 (d, J 8.3 Hz, 1H, H-1GlcN), 4.82–4.79
(m, 2H, PhCH2), 4.61 (d, J 12.2 Hz, 1H, PhCHH),
4.49–4.39 (m, 3H, H-1Man, PhCH2), 4.34–4.13 (m, 3H,
PhCHH, 2,3-HGlcN), 4.03–3.98 (m, 2H, H-4Man, H-
4GlcN), 3.78–3.44 (m, 7H, H-2,3,6Man, H-5,6GlcN),
2.90–2.84 (m, 1H, H-5Man), 2.61 (br, 1H, 2-OHMan),
1.98–1.41 (m, 10H, cyclohexyl), 1.14 (s, 9H, C(CH3)3);
13C NMR (100 MHz, CDCl3): d 167.5, 138.5, 137.6,
137.1, 136.2, 135.8, 133.8, 133.4, 132.6, 131.5, 129.9,
129.7, 128.3, 128.0, 127.8, 127.7, 127.6, 127.4, 126.8,
123.0, 100.2, 99.8, 97.5, 78.8, 74.5, 74.4, 73.4, 73.1,
71.3, 70.8, 69.8, 68.1, 67.6, 61.2, 55.7, 38.0, 27.7, 26.9,
25.6, 22.6, 22.3, 19.3. Anal. Calcd for C63H69NO12Si:
C, 71.36; H, 6.56; N, 1.32. Found: C, 71.33; H, 6.52;
N, 1.25.

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
683
9.5 Hz, 1H, H-4Man), 3.89–3.83 (m, 1H, H-4GlcN), 3.76
(d,
2.9 Hz, 1H, H-2Man), 3.72–3.64 (m, 2H,
10% HFÆpyridine. It was compressed to 1.0 GPa and
left for 12 h. The resulting mixture was diluted with
EtOAc and washed with satd aq NaHCO3 and brine,
successively. The organic layer was dried over MgSO4
and evaporated under diminished pressure. The residue
was purified by silica gel column chromatography (1:1
hexane–EtOAc) to afford 1.10 g (88%) of compound
J
H-3,6aMan), 3.60–3.57 (m, 1H, H-6aGlcN), 3.46 (t,
J 10.4 Hz, 1H, H-6bMan), 3.37–3.29 (m, 2H, H-
5,6bGlcN), 3.13 (dd, J = 6.7, 12.1 Hz, 1H, H-5bXyl),
2.84–2.76 (m, 1H, H-5Man), 2.08 (s, 3H, CH3CO), 2.02
(s, 3H, CH3CO), 1.89 (s, 3H, CH3CO), 1.81–1.25 (m,
10H, cyclohexyl), 1.10 (s, 9H, C(CH3)3); 13C NMR
(100 MHz, CDCl3): d 170, 169.3, 168.8, 167.4, 138.3,
137.6, 137.0, 136.4, 136.0, 134.3, 133.4, 131.4, 129.5,
129.4, 128.3, 128.0, 127.7, 127.4, 127.3, 127.2, 126.8,
123.0, 102.4, 99.8, 98.5, 97.2, 80.4, 75.5, 74.8, 74.5,
73.4, 72.6, 70.7, 70.2, 69.7, 69.5, 68.7, 68.5, 68.3, 61.4,
60.8, 55.6, 38.0, 27.7, 27.0, 26.0, 22.7, 22.4, 21.0, 20.9,
20.8, 19.5; HRESIMS: found m/z 1340.52077
[M+Na]+, calcd for C74H83O19NSiNa 1340.52262.
1
20a; [a]D ꢀ35.3 (c 1.0, CHCl3); H NMR (400 MHz,
CDCl3): d 7.63–6.77 (m, 19H, Ar), 5.18 (t, J 7.6 Hz,
1H, H-3Xyl), 5.07–5.05 (m, 1H, H-1GlcN), 4.96–4.87
(m, 3H, H-1,2,4Xyl), 4.79–4.70 (m, 3H, PhCH2), 4.56
(br s, 1H, H-1Man), 4.49–4.45 (m, 2H, PhCH2), 4.28
(d, J 12.2 Hz, 1H, PhCHH), 4.23–4.17 (m, 3H, H-
2,3GlcN, H-5aXyl), 3.98–3.93 (m, 1H, H-4GlcN), 3.84
(d, J 3.2 Hz, 1H, H-2Man), 3.73–3.72 (m, 2H, H-6GlcN),
3.66–3.56 (m, 2H, H-4,6aMan), 3.52–3.48 (m, 1H, H-
5GlcN), 3.42–3.31 (m, 3H, H-3,6bMan, H-5bXyl), 2.97–
2.91 (m, 1H, H-5Man), 2.87 (d, J 9.3 Hz, 1H, 3-OH-
Man), 2.07 (s, 3H, CH3CO), 2.02 (s, 6H, CH3CO),
1.99–1.32 (m, 10H, cyclohexyl); 13C NMR (100 MHz,
CDCl3): d 169.5, 169.3, 169.0, 167.5, 138.4, 137.4,
136.9, 133.4, 131.4, 128.5, 128.0, 127.9, 127.7, 127.4,
127.3, 126.8, 123.0, 102.1, 99.9, 99.8, 97.3, 80.8, 77.5,
74.6, 74.4, 73.8, 70.8, 70.6, 70.4, 70.1, 68.5, 68.4,
68.2, 61.4, 61.1, 55.6, 38.0, 27.9, 25.7, 22.9, 22.6,
21.2, 20.9, 20.8. Anal. Calcd for C58H65NO19: C,
64.49; H, 6.07; N, 1.30. Found: C, 64.39; H, 5.99; N,
1.26.
3.4. Benzyl (2-O-acetyl-3-O-tert-butyldiphenylsilyl-4,6-
O-cyclohexylidene-b-D-mannopyranosyl)-(1!4)-3,6-di-
O-benzyl-2-deoxy-2-phthalimido-b-D-glucopyranoside
(19b)
To a mixture of 17 (56.2 mg, 53.0 lmol) in pyridine
(1.0 mL) were added Ac2O (0.1 mL) and DMAP (5 mg)
at room temperature and the mixture was stirred for
16 h at the same temperature and evaporated under
diminished pressure. After concentration, the residue
was purified by preparative TLC (4:1 hexane–EtOAc)
to give 56.4 mg (97%) of 19b; [a]D ꢀ3.7 (c 1.0, CHCl3);
1H NMR (400 MHz, CDCl3): d 7.73–6.81 (m, 29H, Ar),
5.01–4.99 (m, 1H, H-1GlcN), 4.94 (d, J 3.4 Hz, 1H, H-
2Man), 4.73 (d, J 12.2 Hz, 1H, PhCHH), 4.72 (d, J
12.2 Hz, 1H, PhCHH), 4.58 (d, J 12.0 Hz, 1H, PhCHH),
4.42 (d, J 12.2 Hz, 1H, PhCHH), 4.38 (br s, 1H, H-1Man),
4.29 (d, J 12.2 Hz, 1H, PhCHH), 4.23 (d, J 12.0 Hz, 1H,
PhCHH), 4.14–4.07 (m, 2H, H-2,3GlcN), 3.98–3.86 (m,
1H, H-4GlcN), 3.89 (t, J 9.6 Hz, 1H, H-4Man), 3.69–3.60
(m, 3H, H-3,6aMan, H-6aGlcN), 3.52–3.44 (m, 2H, H-
6bMan, H-6bGlcN), 3.40–3.38 (br, 1H, H-5GlcN), 2.81–
2.75 (m, 1H, H-5Man), 2.13 (s, 3H, CH3CO), 1.96–1.33
(m, 10H, cyclohexyl), 1.04 (s, 9H, C(CH3)3); 13C NMR
(100 MHz, CDCl3): d 169.6, 167.5, 138.5, 137.8, 137.0,
136.2, 133.7, 133.4, 133.3, 131.5, 129.8, 129.4, 128.3,
128.0, 127.8, 127.7, 127.6, 127.5, 127.4, 127.1, 126.9,
123.0, 99.7, 99.1, 97.2, 78.8, 76.7, 74.4, 73.3, 72.1, 71.6,
70.7, 70.1, 68.1, 67.7, 61.1, 55.6, 37.8, 27.7, 26.8, 25.6,
22.6, 22.4, 21.2, 19.3; HRESIMS: found m/z 1124.46041
[M+Na]+, calcd for C65H71O13NSiNa 1124.45924.
3.6. Benzyl (2-O-acetyl-4,6-O-cyclohexylidene-b-D-
mannopyranosyl)-(1!4)-3,6-di-O-benzyl-2-deoxy-2-
phthalimido-b-D-glucopyranoside (20b)
Compound 19a (2.05 g, 1.86 mmol) was desilylated as
described for compound 20a. Purification by silica gel
column chromatography (7:3!3:2 toluene–EtOAc lin-
ear gradient) afforded 1.43 g (89%) of compound 20b:
[a]D ꢀ3.82 (c 1.0, CHCl3); 1H NMR (400 MHz,
CDCl3): d 7.63–6.80 (m, 19H, Ar), 5.21 (d, J 3.4 Hz,
1H, H-2Man), 5.08–5.02 (m, 1H, H-1GlcN), 4.78–4.74
(m, 3H, PhCH2), 4.64 (br s, 1H, H-1Man), 4.49–4.44
(m, 2H, PhCH2), 4.34 (d, J 12.2 Hz, 1H, PhCHH),
4.20–4.14 (m, 2H, H-2,3GlcN), 4.09–4.05 (m, 1H, H-
4GlcN), 3.81 (dd, J 3.1, 11.1 Hz, 1H, H-6aGlcN), 3.74–
3.67 (m, 3H, H-4,6aMan, H-6bGlcN), 3.51–3.46 (m, 3H,
H-3,6bMan, H-5GlcN), 3.00–2.94 (m, 1H, H-5Man), 2.14
(s, 3H, CH3CO), 2.09 (d, J 3.7 Hz, 1H, 3-OHMan),
1.98–1.36 (m, 10H, cyclohexyl); 13C NMR (100 MHz,
CDCl3): d 170.3, 167.4, 138.4, 137.6, 136.9, 133.4,
131.4, 128.4, 127.9, 127.8, 127.7, 127.5, 127.4, 126.9,
123.0, 99.9, 99.3, 97.2, 79.3, 77.2, 76.8, 74.5, 74.4,
73.5, 71.2, 70.7, 70.2, 70.1, 68.2, 67.8, 61.0, 55.7,
37.9, 28.0, 25.6, 22.8, 22.6, 21.2; HRESIMS: found
m/z 886.33992 [M+Na]+, calcd for C49H53O13NNa
886.34146.
3.5. Benzyl (2,3,4-tri-O-acetyl-b-D-xylopyranosyl)-
(1!2)-(4,6-O-cyclohexylidene-b-D-mannopyranosyl)-
(1!4)-3,6-di-O-benzyl-2-deoxy-2-phthalimido-b-D-gluco-
pyranoside (20a)
To a Teflon reaction vessel was introduced compound
19a (1.40 g, 1.06 mmol) in DMF (4 mL) containing

684
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
3.7. Methyl (3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-3,4,6-tri-O-benzyl-1-thio-a-D-
mannopyranoside (23)
5.26 (d, J 7.8 Hz, 1H, H-1GlcN2), 5.04 (d, J 8.1 Hz, 1H,
H-1GlcN1), 4.96–4.74 (m, 9H, H-1Man2, H-1,2,3Xyl
,
,
,
PhCH2), 4.68–4.32 (m, 12H, H-2,3GlcN2
PhCH2), 4.22–4.11 (m, 5H, H-1Man1
,
H-4Xyl
,
H-2Man2
A mixture of preactivated molecular sieves 4 A (8 g) and
H2,3GlcN1, PhCHH), 4.04–3.93 (m, 3H, PhCH2), 3.89–
˚
AgOTf (3.39 g, 13.2 mmol) in toluene (40 mL) was stir-
red at ꢀ40 ꢁC for 30 min. A mixture of 22 (3.95 g,
6.61 mmol) and 21 (2.46 g, 5.19 mmol) in 1,2-dichloro-
ethane (80 mL) was added dropwise and the reaction
was stirred for 1 h. It was quenched with Et3N (2 mL)
and satd aq NaHCO3. After being stirred for 15 min,
the resulting mixture was diluted with EtOAc, and fil-
tered through Celite. The filtrate was washed with brine,
dried over MgSO4, and evaporated under diminished
pressure. The residue was purified by silica gel column
chromatography (20:1 toluene–EtOAc) to afford 3.42 g
(63%) of 23; [a]D +41.2 (c 1.0, CHCl3); 1H NMR
(400 MHz, CDCl3): d 7.58–7.49 (m, 4H, Phth), 7.34–
6.81 (m, 30H, Ar), 5.27 (d, J 7.8 Hz, 1H, H-1GlcN),
4.86–4.74 (m, 5H, H-1Man, PhCH2), 4.65–4.44 (m, 5H,
PhCH2), 4.39–4.29 (m, 3H, H-2,3GlcN, PhCHH), 4.15
(t, J 2.5 Hz, 1H, H-2Man), 4.01 (d, J 12.0 Hz, 1H,
PhCHH), 3.96 (d, J 12.0 Hz, 1H, PhCHH), 3.82–3.68
(m, 6H, H-4,5,6GlcN, H-3,5Man), 3.46 (t, J 9.1 Hz, 1H,
H-4Man), 3.37 (dd, J 1.8, 10.9 Hz, 1H, H-6aMan), 2.92
(dd, J 6.6, 10.9 Hz, 1H, H-6bMan), 1.90 (s, 3H, SCH3);
13C NMR (100 MHz, CDCl3): d 138.2, 137.9, 137.8,
137.7, 133.3, 131.6, 128.3, 128.2, 128.1, 128.0, 127.9,
127.8, 127.7, 127.5, 127.4, 127.3, 127.2, 127.1, 122.9,
96.4, 82.6, 79.7, 79.0, 77.9, 75.1, 75.0, 74.8, 74.7, 73.6,
72.7, 71.8, 70.7, 69.8, 69.3, 55.8, 13.8; HRESIMS: found
m/z 1064.40541 [M+Na]+, calcd for C63H63O11NNa
1064.40195.
3.72 (m, 8H, H-2Man1, H-3,5Man2, H-4GlcN1, H-4,6GlcN2
,
H-5aXyl), 3.64–3.61 (m, 1H, H-5GlcN2), 3.58–3.50 (m,
2H, H-4,6aMan1), 3.45–3.21 (m, 7H, H-6bMan1, H-
4,6aMan2
, , J 3.1,
H-5,6GlcN1 H-5bXyl), 3.15 (dd,
10.1 Hz, 1H, H-3Man1), 2.74 (dd, J 7.1, 10.5 Hz, 1H,
H-6bMan2), 2.54–2.48 (m, 1H, H-5Man1), 1.94 (s, 3H,
CH3CO), 1.89 (s, 3H, CH3CO), 1.88 (s, 3H, CH3CO),
1.73–1.25 (m, 10H, cyclohexyl); 13C NMR (100 MHz,
CDCl3): d 169.2, 169.1, 168.8, 167.3, 138.7, 138.4,
138.3, 138.1, 137.9, 137.8, 137.7, 137.6, 137.0, 133.4,
133.2, 131.7, 131.4, 128.7, 128.3, 128.2, 128.1, 128.0,
127.9, 127.8, 127.7, 127.6, 127.5, 127.4, 127.3, 127.1,
127.0, 126.7, 123.0, 101.5, 99.3, 99.1, 98.1, 97.3, 96.6,
79.7, 79.6, 79.4, 76.2, 76.1, 75.7, 72.2, 75.1, 74.9, 74.8,
74.6, 74.1, 73.7, 73.2, 72.8, 71.8, 71.7, 70.8, 70.0, 69.7,
69.5, 69.3, 68.2, 67.9, 61.3, 60.5, 55.9, 55.5, 38.0, 27.9,
25.7, 23.5, 22.7, 21.0, 20.9, 20.7. Anal. Calcd for
C
120H124N2O30: C, 69.48; H, 6.03; N, 1.35. Found: C,
69.51; H, 6.01; N, 1.23.
3.9. Benzyl (3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-benzyl-a-D-manno-
pyranosyl)-(1!3)-(2-O-acetyl-4,6-O-cyclohexylidene-b-
D-mannopyranosyl)-(1!4)-3,6-di-O-benzyl-2-deoxy-2-
phthalimido-b-D-glucopyranoside (24b)
Compound 20b (1.12 g, 1.30 mmol) was glycosylated
with 23 as described for 24a. Purification by silica gel
column chromatography (4:1!2:1 hexane–EtOAc then
9:1!8:1 toluene–EtOAc, linear gradient) afforded
1.91 g (79%) of 24b; [a]D ꢀ11.7 (c 1.0, CHCl3); 1H
NMR (400 MHz, CDCl3): d 7.62–6.81 (m, 53H, Ar),
5.23 (d, J 7.6 Hz, 1H, H-1GlcN2), 5.04 (d, J 3.4 Hz,
1H, H-2Man1), 4.99 (d, J 7.8 Hz, 1H, H-1GlcN1), 4.84–
4.68 (m, 6H, H-1Man2, PhCH2), 4.65–4.31 (m, 11H, H-
2,3GlcN2, PhCH2), 4.25 (d, J 12.2 Hz, 1H, PhCHH),
3.8. Benzyl (3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-benzyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4,6-O-cyclohexylidene-b-D-mannopyranos-
yl)-(1!4)-3,6-di-O-benzyl-2-deoxy-2-phthalimido-b-D-
glucopyranoside (24a)
A mixture of preactivated molecular sieves 4 A (6 g),
4.19 (br s, 1H, H-1Man1), 4.14–3.98 (m, 6H, H-2Man2
,
˚
compounds 23 (3.14 g, 3.0 mmol) and 20a (2.16 g,
2.0 mmol), and DTBMP (1.85 g, 9.0 mmol) in toluene
(80 mL) was stirred at room temperature for 30 min.
MeOTf (1.0 mL, 9 mmol) was added and stirred at the
same temperature for 1.5 h. Then the mixture was
warmed up to 50 ꢁC and was stirred for 12 h. The reac-
tion was cooled to ambient temperature, quenched with
Et3N (1.3 mL) and filtered through Celite. The filtrate
was diluted with EtOAc, washed with brine, dried over
MgSO4 and evaporated under diminished pressure.
The residue was purified by silica gel column chroma-
tography (6:1 toluene–EtOAc) to afford 3.90 g (94%)
of compound 24a; [a]D ꢀ21.5 (c 1.0, CHCl3); 1H
NMR (400 MHz, CDCl3): d 7.62–6.79 (m, 53H, Ar),
H-2,3GlcN1, PhCH2), 3.92 (t, J 8.9 Hz, 1H, H-4GlcN1),
3.76–3.72 (m, 3H, H-4,6GlcN2), 3.70–3.58 (m, 4H, H-
6aMan1, H-3,5Man2, H-5GlcN2), 3.56–3.39 (m, 5H, H-
4,6bMan1, H-4Man2, H-6GlcN1), 3.35–3.27 (m, 3H, H-
3Man1, H-6aMan2, H-5GlcN1), 2.72 (dd, J 6.6, 11.0 Hz,
1H, H-6bMan2), 2.64–2.58 (m, 1H, H-5Man1), 1.86 (s,
3H, CH3CO), 1.75–1.23 (m, 10H, cyclohexyl); 13C
NMR (100 MHz, CDCl3): d 169.3, 167.4, 138.7, 138.4,
138.3, 137.9, 137.8, 137.7, 137.6, 136.9, 133.3, 131.6,
131.4, 128.7, 128.6, 128.3, 128.2, 128.1, 128.0, 127.9,
127.8, 127.7, 127.6, 127.5, 127.4, 127.3, 127.1, 127.0,
126.9, 123.0, 99.5, 98.7, 97.7, 97.2, 96.0, 79.6, 79.4,
78.2, 77.2, 76.6, 75.3, 75.0, 74.8, 74.3, 74.0, 73.6,
73.2, 72.5, 72.2, 71.6, 70.8, 70.7, 70.2, 70.1, 69.4, 67.8,

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
685
67.2, 61.2, 55.8, 55.6, 53.5, 38.0, 31.0, 28.1, 25.7,
23.3, 22.8, 20.9. Anal. Calcd for C111H112N2O24: C,
71.75; H, 6.08; N, 1.51. Found: C, 71.78; H, 6.07; N,
1.51.
product was purified by silica gel column chromatogra-
phy (3:1 toluene–EtOAc) to afford 1.52 g (86%) of com-
pound 25b; [a]D +4.6 (c 1.0, CHCl3); 1H NMR
(400 MHz, CDCl3): d 7.64–6.77 (m, 53H, Ar), 5.27 (d,
J 7.8 Hz, 1H, H-1GlcN2), 5.12 (d, J 3.2 Hz, 1H,
H-2Man1), 5.04 (d, J 8.3 Hz, 1H, H-1GlcN1), 4.96 (d,
J = 2.7 Hz, 1H, H-1Man2), 4.84–4.74 (m, 4H, PhCH2),
4.71–4.40 (m, 10H, H-1Man1, PhCH2), 4.36–4.25 (m,
3.10. Benzyl (3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-
b-D-glucopyranosyl)-(1!2)-O-(3,4,6-tri-O-benzyl-a-D-
mannopyranosyl)-(1!3)-[(2,3,4-tri-O-acetyl-b-D-xylo-
pyranosyl)-(1!2)]-(b-D-mannopyranosyl)-(1!4)-3,6-di-
O-benzyl-2-deoxy-2-phthalimido-b-D-glucopyranoside
(25a)
5H, H-2,3GlcN2, PhCH2), 4.17–3.98 (m, 6H, H-2Man2
,
H-2,3,4GlcN1, PhCH2), 3.77–3.75 (br, 1H, H-6aGlcN2),
3.71–3.62 (m, 8H, H-6aMan1, H-3,5Man2, H-6GlcN1, H-
4,5,6bGlcN2), 3.55–3.40 (m, 4H, H-4,6bMan1, H-4Man2
,
To a stirred soln of 24a (29.9 mg, 14.4 lmol) in MeCN
(1 mL) was added TsOHÆH2O (6.8 mg, 36.0 lmol). The
mixture was stirred for 9 h at room temperature and
the reaction was quenched with Et3N (0.1 mL). The
resulting mixture was diluted with EtOAc, washed with
satd aq NaHCO3 and brine, successively, dried over
MgSO4, and evaporated under diminished pressure.
The residue was purified by preparative TLC (1:1 hex-
ane–EtOAc) to afford 26.0 mg (91%) of compound
25a; [a]D ꢀ7.2 (c 1.0, CHCl3); 1H NMR (400 MHz,
CDCl3): d 7.64–6.77 (m, 53H, Ar), 5.34 (d, J 8.3 Hz,
H-5GlcN1), 3.37–3.33 (m, 2H, H-3Man1, H-6aMan2), 3.18
(d, J 4.9 Hz, 1H, 4-OHMan1), 2.91–2.87 (m, 2H, H-
5Man1, H-6bMan2), 1.99 (br, 1H, 6-OHMan1), 1.60 (s,
3H, CH3CO); 13C NMR (100 MHz, CDCl3): d 169.4,
167.4, 138.2, 138.1, 137.9, 137.7, 137.5, 137.0, 133.5,
131.4, 128.5, 128.3, 128.1, 128.0, 127.9, 127.8, 127.7,
127.6, 127.5, 127.4, 127.3, 127.2, 127.1, 123.1, 98.1,
97.2, 96.8, 96.4, 79.6, 79.3, 78.0, 77.9, 77.2, 76.8, 75.4,
75.2, 75.1, 74.8, 74.7, 74.4, 74.3, 74.0, 73.9, 73.6, 73.4,
72.8, 71.4, 71.2, 70.8, 70.7, 69.9, 69.2, 67.9, 66.4, 62.3,
55.8, 55.6, 21.2. Anal. Calcd for C105H104N2O24: C,
70.93; H, 5.90; N, 1.58. Found: C, 70.95; H, 5.86; N,
1.45.
1H, H-1GlcN2), 5.08–4.98 (m, 4H, H-1Man2, H-1GlcN1
,
H-1,3Xyl), 4.92–4.87 (br, 1H, H-4Xyl), 4.85–4.73 (m,
7H, H-2Xyl, PhCH2), 4.59 (d, J 12.2 Hz, 1H, PhCHH),
4.54–4.41 (m, 9H, H-1Man1, H-3GlcN2, PhCH2), 4.39–
4.24 (m, 5H, H-3GlcN1, H-2GlcN2, PhCH2), 4.19–4.14
3.12. Benzyl (3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-
b-D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-benzyl-a-D-
mannopyranosyl)-(1!3)-O-[(2-O-acetyl-3,4,6-tri-O-benz-
yl-a-D-mannopyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-
D-xylopyranosyl)-(1!2)]-(b-D-mannopyranosyl)-(1!4)-
3,6-di-O-benzyl-2-deoxy-2-phthalimido-b-D-glucopyrano-
side (27a)
(m, 2H, H-2Man2, H-2GlcN1), 4.02–3.99 (m, 3H, H-5aXyl
,
PhCH2), 3.85–3.75 (m, 6H, H-2,6aMan1, H-3,5Man2, H-
4GlcN1, H-5GlcN2), 3.69–3.64 (m, 2H, H-4Man2, H-
4GlcN2), 3.61–3.50 (m, 4H, H-4Man1
,
H-6GlcN1
,
H-6aGlcN2), 3.43–3.35 (m, 4H, H-6bMan1, H-6aMan2, H-
5GlcN1, H-6bGlcN2), 3.21–3.15 (m, 2H, H-3Man1, H-
5bXyl), 3.06 (d, J 4.9 Hz, 1H, 4-OHMan1), 2.88–2.81 (m,
2H, H-5Man1, H-6bMan2), 2.17 (br, 1H, 6-OHMan1),
1.96 (s, 3H, CH3CO), 1.95 (s, 3H, CH3CO), 1.93 (s,
3H, CH3CO); 13C NMR (100 MHz, CDCl3): d 169.7,
169.2, 167.5, 138.3, 138.2, 138.1, 137.8, 137.5, 136.9,
133.4, 131.5, 131.4, 128.4, 128.2, 128.1, 128.0, 127.9,
127.8, 127.6, 127.5, 127.4, 127.3, 127.2, 127.1, 126.9,
123.1, 101.1, 100.0, 97.1, 97.0, 96.4, 79.7, 79.1, 79.0,
77.2, 77.1, 76.1, 75.0, 74.8, 74.7, 74.6, 74.3, 73.9, 73.5,
73.5, 73.4, 73.2, 72.9, 71.2, 70.7, 70.5, 70.4, 69.5, 68.6,
68.4, 65.4, 62.2, 61.3, 56.0, 55.6, 31.0, 21.2, 20.9. Anal.
Calcd for C114H116N2O30: C, 68.66; H, 5.86; N, 1.40.
Found: C, 68.37; H, 5.81; N, 1.37.
˚
A mixture of preactivated molecular sieves 4 A (200 mg)
and AgOTf (7.8 mg, 30 lmol) in toluene (1 mL) was stir-
red at 0 ꢁC for 30 min, then cooled to ꢀ30 ꢁC. After
being stirred for 10 min, a mixture of 26 (7.7 mg,
15 lmol) and 25a (25.1 mg, 12.6 lmol) in 1,2-dichloro-
ethane (2.0 mL) was added dropwise and the whole mix-
ture was stirred at ꢀ30 ꢁC for 30 min, then warmed up
to ambient temperature. The mixture was stirred for
2.5 h and the reaction was quenched with Et3N
(20 lL). The resulting mixture was diluted with EtOAc
and filtered through Celite. The filtrate was washed with
satd aq NaHCO3 and brine, successively, dried over
MgSO4 and evaporated under diminished pressure.
The residue was purified by preparative TLC (4:1 tolu-
ene–EtOAc) to afford 23.9 mg (77%) of 27a; [a]D ꢀ3.6
3.11. Benzyl (3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-
b-D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-benzyl-a-D-
mannopyranosyl)-(1!3)-(2-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-3,6-di-O-benzyl-2-deoxy-2-phthalimido-b-D-
glucopyranoside (25b)
1
(c 1.0, CHCl3); H NMR (400 MHz, CDCl3): d 7.63–
6.70 (m, 68H, Ar), 5.33–5.32 (m, 1H, H-2Man3), 5.29
(d, J 8.3 Hz, 1H, H-1GlcN2), 5.11–4.98 (m, 3H, H-1GlcN1
,
H-2,3Xyl), 4.91 (d, J 1.7 Hz, 1H, H-1Man2), 4.88–4.72 (m,
10H, H-1Man3, H-1,4Xyl, PhCH2), 4.67–4.20 (m, 20H, H-
1Man1, H-2Man2, H-3GlcN1, H-2,3-HGlcN2, PhCH2), 4.13
(dd, J 8.4, 10.4 Hz, 1H, H-2GlcN1), 4.07 (d, J 12.2 Hz,
Compound 24b (1.85 g, 996 lmol) was treated with
TsOH as described for the preparation of 25a. The crude

686
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
1H, PhCHH), 4.01 (d, J 12.2 Hz, 1H, PhCHH), 3.92–
3.14. (3,4,6-Tri-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4-O-acetyl-b-D-mannopyranosyl)-(1!4)-
1,3,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyra-
nose (28b)
3.68 (m, 15H, H-2,6aMan1, H-3,4,5Man2, H-3,4,5,6Man3
,
H-4GlcN1, H-4,5,6aGlcN2, H-5aXyl), 3.54–3.47 (m, 3H,
H-4Man1, H-6GlcN1), 3.43–3.32 (m, 4H, H-6bMan1, H-
6aMan2
, ,
H-5GlcN1 H-6bGlcN2), 3.17–3.09 (m, 3H,
H-3Man1, 4-OHMan1, H-5bXyl), 2.93–2.85 (m, 2H, H-
5Man1, H-6bMan2), 2.08 (s, 3H, CH3CO), 1.95 (s, 6H,
CH3CO), 1.90 (s, 3H, CH3CO); 13C NMR (100 MHz,
CDCl3): d 170.0, 169.7, 169.1, 168.9, 167.4, 138.6,
138.2, 138.1, 137.9, 137.8, 137.0, 133.3, 131.4, 128.4,
128.2, 128.1, 127.9, 127.8, 127.7, 127.6, 127.5, 127.4,
127.2, 127.1, 127.0, 126.9, 123.0, 101.7, 100.3, 97.8,
97.5, 97.1, 96.6, 80.0, 79.5, 79.2, 78.3, 77.4, 77.2, 76.3,
75.2, 75.0, 74.8, 74.6, 74.2, 74.1, 73.4, 73.2, 72.8, 71.7,
71.6, 71.5, 71.3, 70.7, 70.6, 70.5, 69.2, 69.1, 68.6, 67.5,
66.3, 61.8, 56.0, 55.6, 31.0, 21.2, 21.0, 20.9; HRESIMS:
found m/z 2489.95692 [M+Na]+, calcd for
To a stirred soln of 27a (247 mg, 100 lmol) in cyclo-
hexene (20 mL), EtOH (10 mL), and AcOH (10 mL) was
added 20% Pd(OH)2 (250 mg). The mixture was heated
under reflux for 60 h and filtered through Celite. The fil-
trate was evaporated under diminished pressure, and the
residue was dissolved in pyridine (10 mL) and Ac2O
(5 mL). The mixture was stirred for 6 h and quenched
with EtOH (10 mL) at ice-cold temperature. The result-
ing mixture was evaporated, diluted with EtOAc,
washed with 1 M HCl and brine, successively, dried over
MgSO4, and evaporated under diminished pressure. The
residue was purified by preparative TLC (1:4 hexane–
EtOAc) to afford 180 mg (93%) of 28b; [a]D ꢀ17.0 (c
C
143H146O36N2Na 2489.95529.
3.13. Benzyl (3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-
b-D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-benzyl-a-D-
mannopyranosyl)-(1!3)-[(2-O-acetyl-3,4,6-tri-O-benzyl-
a-D-mannopyranosyl)-(1!6)]-(2-O-acetyl-b-D-mannopyr-
anosyl)-(1!4)-3,6-di-O-benzyl-2-deoxy-2-phthalimido-b-
D-glucopyranoside (27b)
1
1.0, CHCl3); H NMR (400 MHz, CDCl3): d 7.84–7.70
(m, 8H, Ar), 6.44 (d, J 9.0 Hz, 1H), 5.87 (t, J 9.0 Hz,
1H), 5.66 (dd, J 9.0, 10.5 Hz, 1H), 5.31–5.28 (m, 3H),
5.19–5.02 (m, 5H), 4.98–4.91 (m, 2H), 4.79–4.76 (m,
3H), 4.63 (br s, 1H), 4.48 (br s, 1H), 4.42–4.24 (m,
7H), 4.08–3.82 (m, 9H), 3.76–3.68 (m, 3H), 3.57–3.42
(m, 4H), 2.26 (s, 3H, CH3CO), 2.16–1.94 (m, 42H,
CH3CO), 1.88 (s, 3H, CH3CO), 1.84 (s, 3H, CH3CO);
13C NMR (100 MHz, CDCl3): d 170.5 · 2, 170.4,
170.0, 169.8 · 2, 169.7, 169.5, 169.4, 169.2, 169.1,
168.9, 168.3, 167.2, 134.3, 131.2, 123.5, 99.5, 98.8,
98.2, 97.2, 97.0, 89.6, 74.1, 73.3, 72.5, 72.1, 70.6, 69.6,
69.2, 68.8, 68.7, 68.6, 68.0, 67.8, 65.8, 65.5, 62.3, 62.1,
62.0, 61.8, 54.3, 53.4, 21.0, 20.9, 20.8, 20.7, 20.6 · 2,
20.5 · 2, 20.4; HRESIMS: found m/z 1955.52682
[M+Na]+; calcd for C85H100O49N2Na 1955.52923.
Compound 25a (1.48 g, 832 lmol) was glycosylated with
chloride 26 as described for the preparation of com-
pound 27a. The crude product was purified by silica
gel column chromatography (5:2!1:1 hexane–EtOAc,
linear gradient) to afford 1.67 g (89%) of 27b; [a]D
1
+14.6 (c 1.0, CHCl3); H NMR (400 MHz, CDCl3): d
7.65–6.69 (m, 68H, Ar), 5.30–5.29 (m, 1H, H-2Man3),
5.27 (d, J 7.8 Hz, 1H, H-1GlcN2), 5.13 (d, J 3.2 Hz, 1H,
H-2Man1), 4.98 (d, J 8.3 Hz, 1H, H-1GlcN1), 4.95 (d, J
2.0 Hz, 1H, H-1Man2), 4.84 (d, J 1.7 Hz, 1H, H-1Man3),
4.82–4.70 (m, 7H, PhCH2), 4.64–4.57 (m, 3H, PhCH2),
4.52–4.25 (m, 14H, H-1Man1, H-2,3GlcN2, PhCH2), 4.22
(d, J 11.0 Hz, 1H, PhCHH), 4.16 (br s, 1H, H-2Man2),
4.13–4.03 (m, 4H, H-2,3-HGlcN1, PhCH2), 3.93 (br,
3.15. (3,4,6-Tri-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-(2,4-di-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-1,3,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl (28c)
1H, H-4GlcN1), 3.83–3.77 (m, 4H, H-5Man2
3,4,6aMan3), 3.71–3.55 (m, 12H, H-4,6aMan1
,
,
H-
H-
2,3,5Man2, H-5,6bMan3, H-6GlcN1, H-4,5,6aGlcN2), 3.47
(br, 1H, H-6bGlcN2), 3.38–3.30 (m, 3H, H-5GlcN1, H-
3,6bMan1), 3.17 (d, J 5.1 Hz, 1H, 4-OHMan1), 2.94–2.89
(m, 2H, H-5Man1, H-6bMan2), 1.95 (s, 3H, CH3CO),
1.94 (s, 3H, CH3CO); 13C NMR (100 MHz, CDCl3): d
169.8, 169.5, 167.2, 138.4, 138.3, 138.2, 138.1, 138.0,
137.8, 137.7, 133.3, 131.5, 128.5, 128.3, 128.2, 128.1,
128.0, 127.9, 127.8, 127.7, 127.5, 127.4, 127.3, 127.2,
127.1, 126.9, 122.9, 98.7, 97.9, 97.8, 97.1, 96.5, 79.6,
79.3, 78.6, 78.3, 77.2, 76.4, 75.2, 75.1, 74.9, 74.7, 74.3,
74.2, 73.5, 73.4, 73.2, 72.7, 71.7, 71.6, 71.1, 70.6, 69.9,
69.1, 68.8, 68.4, 68.1, 67.2, 66.6, 55.8, 55.7, 21.0; HRE-
SIMS: found m/z 2273.89419 [M+Na]+; calcd for
Compound 27a (226 mg, 100 lmol) was submitted to
sequential debenzylation and acetylation as described
for 28b. Purification by preparative TLC (1:5 hexane–
EtOAc) afforded 166 mg (97%) of 28c; [a]D +5.5 (c
1
1.0, CHCl3); H NMR (400 MHz, CDCl3): d 7.82–7.70
(m, 8H, Ar), 6.41 (d, J 8.8 Hz, 1H), 5.87 (dd, J 8.3,
10.7 Hz, 1H), 5.64 (t, J 9.8 Hz, 1H), 5.40–5.22 (m,
5H), 5.14–5.09 (m, 2H), 5.02 (t, J 9.5 Hz, 1H), 4.82 (br
s, 1H), 4.75 (dd, J 3.4, 10.5 Hz, 1H), 4.59–4.57 (m,
2H), 4.40–4.28 (m, 6H), 4.17–4.02 (m, 4H), 3.97–3.90
(m, 2H), 3.86–3.71 (m, 6H), 3.58–3.47 (m, 2H), 2.30 (s,
C
134H134O30N2Na 2273.89191.

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
687
3H, CH3CO), 2.17–1.92 (m, 39H, CH3CO), 1.66 (s, 3H,
CH3CO); 13C NMR (100 MHz, CDCl3): d 170.5 · 2,
170.2, 170.0 · 2, 169.9, 169.6 · 2, 169.2, 169.1, 168.2,
167.1, 134.3, 131.1, 123.4, 98.3, 97.5, 97.0, 96.7, 89.7,
74.0, 73.9 · 2, 73.1, 72.8, 72.1, 69.3, 69.2 · 3, 69.0,
68.8, 68.6, 67.6, 65.7, 64.9, 62.3, 62.1, 62.0, 61.8, 54.3,
53.4, 21.0, 20.9, 20.8, 20.7 · 3, 20.6, 20.5, 20.4 · 2; HRE-
SIMS: found m/z 1739.47052 [M+Na]+; calcd for
C76H88O43N2Na 1739.46585.
CDCl3): d 8.62 (s, 1H, NH), 7.81–7.66 (m, 8H, Ar),
6.53 (d, J 9.0 Hz, 1H), 5.89 (dd, J 9.0, 10.7 Hz, 1H),
5.63 (t, J 9.5 Hz, 1H), 5.39 (t, J 10.0 Hz, 1H), 5.31 (dd,
J 3.2, 10.0 Hz, 1H), 5.24–5.22 (m, 3H), 5.13–5.07 (m,
2H), 5.00 (t, J 9.5 Hz, 1H), 4.81 (br s, 1H), 4.74 (dd, J
3.2, 10.0 Hz, 1H), 4.59–4.57 (m, 2H), 4.46–4.26 (m,
6H), 4.18–3.92 (m, 6H), 3.85–3.70 (m, 6H), 3.56–3.47
(m, 2H), 2.29 (s, 3H, CH3CO), 2.15–1.93 (m, 36H,
CH3CO), 1.82 (s, 3H, CH3CO); 13C NMR (100 MHz,
CDCl3): d 170.5 · 2, 170.2, 170.0 · 2, 169.9, 169.8,
169.7, 169.6, 169.2 · 2, 160.2, 134.4, 134.3, 131.2, 131.0,
123.6, 123.5, 98.4, 97.5, 97.0, 96.8, 93.6, 90.1, 74.1, 73.8,
73.4, 73.0, 72.1, 70.6, 69.3 · 2, 69.2 · 2, 69.1 · 2, 68.8,
68.6, 67.6, 65.6, 65.0, 62.3, 62.2, 61.9, 61.8, 54.3, 53.4,
21.1, 21.0, 20.9, 20.8 · 2, 20.7 · 2, 20.6, 20.5 · 2, 20.4;
HRESIMS: found m/z 1840.36317 [M+Na]+; calcd for
C76H86O42N3Cl3Na 1840.36491.
3.16. (3,4,6-Tri-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4-O-acetyl-b-D-mannopyranosyl)-(1!4)-
3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyranos-
yl trichloroacetimidate (9)
To a stirred soln of 28b (43.8 mg, 22.7 lmol) in DMF
(3 mL) was added hydrazine acetate (2.7 mg, 30 lmol).
The mixture was stirred for 2 h, washed with brine, dried
over MgSO4, and evaporated under diminished pres-
sure. The residue was dissolved in trichloroacetonitrile
(5 mL). Then DBU (3.2 lL, 22 lmol) was added and
the reaction was stirred for 12 h. The resulting mixture
was evaporated under diminished pressure and the resi-
due was purified by preparative TLC (1:4 hexane–
EtOAc containing 1% Et3N) to afford 36.4 mg (79%)
3.18. Benzyl 4,6-O-benzylidene-2-deoxy-3-O-p-meth-
oxybenzyl-2-phthalimido-b-D-glucopyranoside (32)
To a stirred soln of 31 (975 mg, 2.0 mmol) in CH2Cl2
(10 mL) were added PMB–OC(NH)CCl3 (5.0 mmol) in
CH2Cl2 (10 mL), and La(OTf)3 (117 mg, 0.2 mmol), suc-
cessively, and stirring was continued for 17 h. Addi-
tional amounts of PMB–OC(NH)CCl3 (1.0 mmol) in
CH2Cl2 (2 mL), and La(OTf)3 (23.4 mg, 0.04 mmol)
were added. The mixture was stirred for 3 h and
quenched with Et3N (2 mL). The resulting mixture was
diluted with EtOAc, washed with satd aq NaHCO3
and brine, successively, dried over MgSO4, and evapo-
rated under diminished pressure. The residue was puri-
fied by silica gel column chromatography (5:1!4:1
hexane–EtOAc) and recrystallization from 2-propanol
to afford 954 mg (79%) of 32; [a]D +10.4 (c 1.0, CHCl3);
1
of 9; [a]D ꢀ13.5 (c 1.0, CHCl3); H NMR (400 MHz,
CDCl3): d 8.62 (s, 1H, NH), 7.83–7.64 (m, 8H, Ar),
6.54 (d, J 8.8 Hz, 1H), 5.88 (dd, J 8.8, 10.5 Hz, 1H),
5.64 (dd, J 9.0, 10.5 Hz, 1H), 5.33–5.26 (m, 3H), 5.18–
5.01 (m, 5H), 4.96–4.92 (m, 2H), 4.77–4.73 (m, 3H),
4.62 (br s, 1H), 4.50–4.45 (m, 2H), 4.41–4.22 (m, 6H),
4.11–3.81 (m, 9H), 3.75–3.68 (m, 3H), 3.57–3.43 (m,
4H), 2.24 (s, 3H, CH3CO), 2.14–1.82 (m, 45H); 13C
NMR (100 MHz, CDCl3): d 170.5, 170.4, 170.0, 169.9,
169.8, 169.7, 169.5 · 2, 169.4, 169.2, 169.1, 168.9,
160.3, 134.3, 131.2, 131.1, 123.5, 99.6, 98.8, 98.2, 97.3,
97.0, 93.4, 90.1, 74.1, 73.5, 72.7, 72.1, 70.6, 69.7, 69.2,
68.9 · 2, 68.8, 68.7, 68.2, 67.8, 65.8, 65.6, 62.3, 62.2,
61.9 · 2, 54.4, 53.5, 21.1, 21.0, 20.9, 20.8, 20.7,
20.6 · 2, 20.5, 20.4; HRESIMS: found m/z 2056.43055
[M+Na]+; calcd for C85H98O48N3Cl3Na 2056.42830.
1
mp 125–127 ꢁC; H NMR (400 MHz, CDCl3): d 7.66
(br, 3H, Ar), 7.52–7.50 (m, 3H, Ar), 7.40–7.35 (m, 3H,
Ar), 7.03–6.97 (m, 4H, Ar), 6.87 (d, J 8.5 Hz, 2H, Ar),
6.33 (d, J 8.5 Hz, 2H, Ar), 5.61 (s, 1H, benzylidene-H),
5.18 (d, J 8.5 Hz, 1H, H-1GlcN), 4.68 (d, J 12.4 Hz,
1H, ArCHH), 4.68 (d, J 12.2 Hz, 1H, ArCHH), 4.47–
4.34 (m, 4H, ArCH2, H-3,6aGlcN), 4.20 (dd, J 8.4,
10.3 Hz, 1H, H-2GlcN), 3.86 (t, J 10.3 Hz, 1H, H-
6bGlcN), 3.79 (t, J 9.3 Hz, 1H, H-4GlcN), 3.64–3.56 (m,
4H, H-5GlcN, OCH3); 13C NMR (100 MHz, CDCl3): d
167.3, 158.7, 137.2, 136.8, 133.5, 131.5, 130.0, 129.6,
128.9, 128.2, 128.1, 127.6, 127.5, 126.0, 123.0, 113.2,
101.3, 97.8, 83.0, 74.0, 73.6, 71.1, 68.8, 66.1, 55.9, 54.9;
HRESIMS: found m/z 630.20998 [M+Na]+; calcd for
C36H33O8NNa 630.21039.
3.17. (3,4,6-Tri-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-(2,4-di-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl trichloroacetimidate (12)
3.19. Benzyl 2-deoxy-3-O-p-methoxybenzyl-2-phthal-
imido-b-D-glucopyranoside (33)
Compound 28c (56.9 mg, 33.1 lmol) was submitted to
anomeric deacetylation and trichloroacetimidate forma-
tion as described for 9 to afford 44.6 mg (74%, two steps)
To a stirred soln of 32 (27.2 mg, 44.8 lmol) in MeCN
(1 mL) and MeOH (1 mL), was added TsOHÆH2O
1
of 12; [a]D +10.4 (c 1.0, CHCl3); H NMR (400 MHz,

688
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
(29.9 mg, 157 lmol). The reaction mixture was stirred
for 11 h at room temperature and was then quenched
with Et3N (50 lL). The resulting mixture was diluted
with EtOAc, washed with satd aq NaHCO3 and brine,
successively, dried over MgSO4, and evaporated under
diminished pressure. The residue was purified by pre-
parative TLC (1:2 hexane–EtOAc) to afford 18.2 mg
(78%) of 33; [a]D ꢀ4.8 (c 1.0, CHCl3); 1H NMR
(400 MHz, CDCl3): d 7.78–7.57 (br, 4H, Ar), 7.10–6.94
(m, 7H, Ar), 6.50–6.47 (m, 2H, Ar), 5.17 (d, J 8.3 Hz,
1H, H-1GlcN), 4.74 (d, J 12.2 Hz, 1H, ArCHH), 4.55
(d, J 12.2 Hz, 1H, ArCHH), 4.48 (d, J 12.2 Hz, 1H,
ArCHH), 4.42 (d, J 12.2 Hz, 1H, ArCHH), 4.22 (dd, J
8.3, 10.5 Hz, 1H, H-3GlcN), 4.14 (dd, J 8.3, 10.5 Hz,
1H, H-2GlcN), 3.95–3.90 (m, 1H, H-6aGlcN), 3.86–3.79
(m, 1H, H-6bGlcN), 3.76–3.70 (m, 1H, H-4GlcN), 3.61
(s, 3H, OCH3), 3.52–3.48 (m, 1H, H-5GlcN), 2.48 (d, J
3.7 Hz, 1H, 4-OHGlcN), 2.05 (t, J 6.5 Hz, 1H, 6-OHGlcN);
13C NMR (100 MHz, CDCl3): d 171.7, 158.8, 137.0,
133.6, 131.5, 130.1, 129.4, 128.1, 127.6, 123.1, 113.5,
97.6, 78.7, 75.2, 74.0, 72.1, 71.2, 62.4, 55.6, 54.9; HRE-
SIMS: found m/z 542.17507 [M+Na]+; calcd for
C29H29O8NNa 542.17909.
CH3CO), 1.99 (s, 3H, CH3CO), 1.10 (d, J 6.6 Hz, 3H,
6-CH3Fuc); 13C NMR (100 MHz, CDCl3): d 170.3,
169.8, 158.6, 137.6, 137.0, 133.5, 131.5, 130.4, 129.5,
128.4, 128.0, 127.9, 127.5, 127.4, 123.1, 122.9, 113.3,
98.0, 97.2, 77.7, 73.8, 73.7, 73.6, 73.4, 73.3, 71.6, 70.6,
70.3, 68.5, 64.8, 55.5, 54.9, 20.9, 20.8, 15.9. Anal. Calcd
for C46H49NO14: C, 65.78; H, 5.88; N, 1.67. Found: C,
65.42; H, 5.70; N, 1.60.
3.21. Benzyl 6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl-
2-phthalimido-b-D-glucopyranoside (13)
To a stirred soln of compound 32 (30.4 mg, 50.0 lmol)
in THF (2 mL) was added BH3ÆNMe3 (18.2 mg,
250 lmol) at room temperature, then AlCl3 (33.3 mg,
250 lmol) was added at ice-cold temperature. After
being stirred for 20 h at room temperature, the mixture
was quenched with water (2 mL) and 1 M HCl (5 mL).
The resulting mixture was diluted with EtOAc, washed
with brine, dried over MgSO4, and evaporated under
diminished pressure. The residue was purified by pre-
parative TLC (1:1 hexane–EtOAc) to afford 20.0 mg
(66%) of 13; [a]D ꢀ7.8 (c 1.0, CHCl3); 1H NMR
(400 MHz, CDCl3): d 7.77–6.99 (m, 14H, Ar), 6.94 (d,
J 8.5 Hz, 2H, Ar), 6.42 (d, J 8.5 Hz, 2H, Ar), 5.14 (d,
J 7.8 Hz, 1H, H-1GlcN), 4.77 (d, J 12.4 Hz, 1H,
ArCHH), 4.67–4.57 (m, 3H, ArCH2), 4.47–4.42 (m,
2H, ArCH2), 4.23–4.14 (m, 2H, H-2,3GlcN), 3.86–3.76
(m, 3H, H-4,6GlcN), 3.65–3.60 (m, 1H, H-5GlcN), 3.59
(s, 3H, OCH3), 2.93 (d, J 2.7 Hz, 1H, 4-OHGlcN); 13C
NMR (100 MHz, CDCl3): d 167.7, 158.6, 137.6, 137.0,
133.5, 131.5, 130.2, 129.4, 128.4, 128.0, 127.8, 127.7,
127.5, 127.4, 123.1, 122.9, 113.3, 97.3, 78.2, 74.2, 73.8,
73.7, 73.6, 70.7, 70.6, 55.4, 54.8; HRESIMS: found
m/z 623.22545 [M+Na]+; calcd for C36H35O8NNa
623.22604.
3.20. Benzyl (3,4-di-O-acetyl-2-O-benzyl-a-L-fucopyr-
anosyl)-(1!6)-2-deoxy-3-O-p-methoxybenzyl-2-phthal-
imido-b-D-glucopyranoside (10)
˚
A
mixture of preactivated molecular sieves 4 A
(800 mg), 11 (168 mg, 455 lmol) and 33 (169 mg,
325 lmol), and DTBMP (281 mg, 1.37 mmol) in cyclo-
pentyl methyl ether (CPME) (15 mL) was stirred at
room temperature for 30 min. MeOTf (1 M 1,2-dichlo-
roethane soln, 1.4 mL) was added. After being stirred
for 8 h at room temperature, 11 (36.0 mg, 97.5 lmol)
in CPME (1 mL), and MeOTf (1 M 1,2-dichloroethane
soln, 290 lL) were added, successively, and stirred for
14 h at the same temperature. The reaction was
quenched with Et3N (1 mL) and filtered through Celite.
The filtrate was diluted with EtOAc, washed with brine,
dried over MgSO4, and evaporated under diminished
pressure. The residue was purified by silica gel column
chromatography (3:1!2:1 hexane–EtOAc, linear gradi-
ent) to afford 179 mg (66%) of 10; [a]D ꢀ65.2 (c 1.0,
3.22. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4-O-acetyl-b-D-mannopyranosyl)-(1!4)-
(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyrano-
syl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-fucopyranos-
yl)-(1!6)]-2-deoxy-3-O-p-methoxybenzyl-2-phthalimido-
b-D-glucopyranoside (34)
1
CHCl3); H NMR (400 MHz, CDCl3): d 7.76–7.50 (br,
4H, Ar), 7.35–7.25 (m, 5H, Ar), 7.06–6.99 (m, 5H,
Ar), 6.90 (d, J 8.5 Hz, 2H, Ar), 6.40 (d, J 8.5 Hz, 2H,
Ar), 5.33 (dd, J 3.4, 10.2 Hz, 1H, H-3Fuc), 5.30–5.29
(m, 1H, H-4Fuc), 5.10–5.08 (m, 1H, H-1GlcN), 4.88 (d,
J 3.7 Hz, 1H, H-1Fuc), 4.72 (d, J 12.2 Hz, 2H, ArCH2),
4.63–4.57 (m, 2H, ArCH2), 4.41 (d, J 12.2 Hz, 1H,
ArCHH), 4.38 (d, J 12.2 Hz, 1H, ArCHH), 4.26–4.21
(br, 1H, H-5Fuc), 4.17–4.13 (m, 2H, H-2,3-HGlcN), 3.95
(dd, J 4.2, 11.2 Hz, 1H, H-6aGlcN), 3.88–3.83 (m, 3H,
˚
A mixture of preactivated molecular sieves 4 A (200 mg)
and compounds 9 (44.1 mg, 21.7 lmol) and 10 (36.5 mg,
43.4 lmol) in CH2Cl2 (2 mL) was stirred at ꢀ78 ꢁC for
30 min. TMSOTf (0.05 M CH2Cl2 soln, 87 lL) was
added and the reaction was stirred at ꢀ40 ꢁC for 4.5 h.
The reaction was quenched with Et3N (50 lL) and fil-
tered through Celite. The filtrate was diluted with
EtOAc, washed with satd aq NaHCO3 and brine, suc-
cessively, dried over MgSO4, and evaporated under
H-4,6bGlcN
, ,
H-2Fuc), 3.60–3.54 (m, 4H, H-5GlcN
OCH3), 3.33 (d, J 3.4 Hz, 1H, 4-OHGlcN), 2.14 (s, 3H,

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
689
diminished pressure. The residue was purified by pre-
parative TLC (1:3 hexane–EtOAc) to afford 43.8 mg
(74%) of 34 and 19.7 mg (54% recovery) of 10. Com-
pound 34: 1H NMR (400 MHz, CDCl3): d 7.84–6.81
(m, 24H, Ar), 6.22 (d, J 8.3 Hz, 2H, Ar), 5.81 (dd, J
9.3, 10.8 Hz, 1H), 5.66 (dd, J 9.3, 10.8 Hz, 1H), 5.52
(d, J 8.3 Hz, 1H), 5.29–4.85 (m, 15H), 4.76–4.22 (m,
17H), 4.15–3.62 (m, 19H), 3.55–3.29 (m, 8H), 2.23 (s,
3H, CH3CO), 2.11–1.84 (m, 51H, CH3CO), 1.03 (d, J
6.3 Hz, 3H, 6-CH3Fuc); 13C NMR (100 MHz, CDCl3): d
170.5, 170.4, 170.3, 170.2, 169.9, 169.8, 169.7, 169.6,
169.5, 169.4, 169.3, 169.1, 169.0, 168.8, 168.0, 167.1,
158.2, 138.0, 136.8, 134.6, 134.3, 133.1, 131.4, 131.2,
131.1, 130.8, 129.5, 128.3, 127.9, 127.6, 127.3, 127.2,
123.7, 123.4, 122.6, 113.0, 99.5, 98.6, 98.0, 97.4, 97.0,
96.9, 96.8, 96.5, 76.2, 75.6, 74.1, 73.9, 72.6, 72.3, 72.2,
72.1, 71.7, 70.7, 70.1, 70.0, 69.5, 69.3, 68.9, 68.7, 68.5,
67.9, 65.8, 65.6, 65.4, 64.2, 62.3, 62.1, 62.0, 61.9, 55.7,
55.1, 54.7, 54.4, 21.1, 21.0, 20.9, 20.8, 20.7, 20.6, 16.0;
HRESIMS: found m/z 2734.82033 [M+Na]+; calcd for
97.2, 97.0, 96.9, 77.1, 74.2, 73.0, 72.6, 72.3, 72.2, 72.1,
71.4, 70.7, 70.4, 69.9, 69.6, 69.5, 69.3, 69.1, 69.0, 68.8,
68.5, 68.4, 67.9, 67.7, 65.7, 65.5, 64.2, 62.4, 62.3, 62.2,
61.9, 55.9, 54.5, 54.4, 21.1, 21.0, 20.9, 20.8, 20.7, 20.6,
20.5, 16.0; HRESIMS: found m/z 2614.76628
[M+Na]+; calcd for C121H137O60N3Na 2614.76589.
3.24. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4-O-acetyl-b-D-mannopyranosyl)-(1!4)-
(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyrano-
syl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-fucopyrano-
syl)-(1!6)]-[(3,4-di-O-acetyl-2-O-benzyl-a-L-fucopyrano-
syl)-(1!3)]-2-deoxy-2-phthalimido-b-D-glucopyranoside
(36)
To a mixture of 11 (4.3 mg, 12 lmol) and 35 (10.1 mg,
3.89 lmol), DTBMP (3.6 mg, 18 lmol) and preacti-
˚
C
129H145O61N3Na 2734.82341.
vated MS 4 A (0.25 g) in dry p-xylene (5 mL) was
added MeOTf (35.4 lL, 167 lmol) at room tempera-
3.23. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4-O-acetyl-b-D-mannopyranosyl)-(1!4)-
(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyrano-
syl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-fucopyrano-
syl)-(1!6)]-2-deoxy-2-phthalimido-b-D-glucopyranoside
(35)
ture. The mixture was rapidly mixed and frozen by
liquid nitrogen. The mixture was stored in
a
refrigerator at 4 ꢁC for 84 h and defrosted at room tem-
perature. The reaction was quenched with Et3N and the
mixture was filtrated through Celite. The filtrate was di-
luted with EtOAc, washed with brine, dried over
Na2SO4, and evaporated under diminished pressure.
After gel filtration (Bio-Beads SX-3, 1:1 toluene–
EtOAc), to a mixture of the residue, 11 (4.3 mg,
12 lmol), DTBMP (3.6 mg, 17.5 lmol) and preacti-
˚
To a mixture of 33 (13.9 mg, 5.12 lmol) in dry CH2Cl2
(1.5 mL) were added Mn(OAc)3Æ2H2O (4.9 mg, 18 lmol)
and DDQ (1.4 mg, 6.1 lmol) successively at room tem-
perature. After being stirred for 48 h at the same temper-
ature, the reaction was quenched with satd aq NaHCO3
and diluted with EtOAc. The organic layer was washed
with an aq soln of ascorbic acid (0.7%)–citric acid
(1.3%)–NaOH (0.9%) and brine, successively, dried over
Na2SO4, and evaporated under diminished pressure.
The residue was purified by preparative TLC (1:2 tolu-
vated MS 4 A (0.25 g) in dry p-xylene (5 mL) was
added MeOTf (35.4 lL, 167 lmol) at room tempera-
ture. The mixture was rapidly mixed and frozen by
liquid nitrogen. The mixture was stored in
a
refrigerator at 4 ꢁC for 84 h and defrosted at room tem-
perature. The reaction was quenched with Et3N and the
mixture was filtrated through Celite. The filtrate was
diluted with EtOAc, washed with brine, dried over
Na2SO4 and evaporated under diminished pressure.
The residue was purified by gel filtration (Bio-Beads
SX-3, 1:1 toluene–EtOAc), then by preparative TLC
(1:2 hexane–EtOAc) to afford 10.2 mg (90%) of 36;
1H NMR (400 MHz, CDCl3): d 7.83–6.85 (m, 27H,
Ar), 5.76–5.71 (m, 1H), 5.68–5.63 (m, 1H), 5.59 (d, J
8.3 Hz, 1H), 5.32–4.53 (m, 26H), 4.41–3.25 (m, 35H),
2.22–1.79 (m, 60H, CH3CO), 1.11 (d, J 6.3 Hz, 3H,
6-CHF3 uc), 1.03 (d, J 6.3 Hz, 3H, 6-CHF3 uc); 13C NMR
(100 MHz, CDCl3): d 170.6, 170.5, 170.4, 170.3.
170.2, 169.9, 169.7, 169.6, 169.5, 169.4, 169.3, 169.1,
169.0, 168.8, 167.9, 166.8, 137.9, 137.8, 136.6, 134.4,
133.7, 131.5, 131.3, 131.2, 131.1, 128.3, 128.2, 128.0,
127.8, 127.7, 127.6, 127.4, 127.3, 127.2, 123.6, 123.4,
100.6, 98.8, 98.3, 98.0, 97.4, 96.9, 96.4, 96.3, 95.8,
77.6, 76.4, 74.2, 74.0, 73.8, 73.0, 72.6, 72.2, 72.1, 71.8,
1
ene–EtOAc) to afford 12.9 mg (97%) of 35; H NMR
(400 MHz, CDCl3): d 7.84–6.94 (m, 22H, Ar), 5.83–
5.78 (m, 1H), 5.66 (dd, J 9.0, 10.7 Hz, 1H), 5.38 (d, J
8.3 Hz, 1H), 5.32–5.26 (m, 3H), 5.16–5.06 (m, 6H),
5.02 (t, J 5.4 Hz, 1H), 4.98–4.92 (m, 2H), 4.87–4.85
(m, 2H), 4.79 (d, J 1.2 Hz, 1H), 4.73–4.71 (m, 2H),
4.65–4.57 (m, 4H), 4.43–4.21 (m, 10H), 4.14–3.63 (m,
17H), 3.52–3.29 (m, 7H), 2.25 (s, 3H, CH3CO), 2.16 (s,
3H, CH3CO), 2.09–1.84 (m, 48H, CH3CO), 0.98 (d, J
6.3 Hz, 3H, 6-CH3Fuc); 13C NMR (100 MHz, CDCl3): d
170.5, 170.4, 170.3, 170.2, 170.0, 169.9, 169.8, 169.6,
169.4, 169.3, 169.1, 169.0, 168.7, 167.7, 138.1, 136.8,
134.6, 134.3, 133.7, 131.6, 131.2, 128.4, 128.0, 127.6,
127.4, 127.3, 127.2, 123.4, 98.7, 98.5, 98.2, 98.0, 97.3,

690
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
71.7, 70.7, 70.5, 70.2, 70.0, 69.3, 69.1, 69.0, 68.9, 68.8,
68.7, 68.6, 68.3, 65.7, 65.6, 65.4, 64.6, 64.3, 62.6, 62.3,
62.1, 61.9, 60.0, 56.5, 55.1, 54.4, 21.1, 21.0, 20.9,
20.8, 20.7, 20.6, 20.5, 16.0; HRESIMS: found m/z
2934.88712 [M+Na]+; calcd for C138H157O66N3Na
2934.89188.
3.26. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4-O-acetyl-b-D-mannopyranosyl)-(1!4)-
(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyrano-
syl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-fucopyrano-
syl)-(1!3)]-2-deoxy-2-phthalimido-b-D-glucopyranoside
(38)
3.25. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-[(2,3,4-tri-O-acetyl-b-D-xylopyranos-
yl)-(1!2)]-(4-O-acetyl-b-D-mannopyranosyl)-(1!4)-
(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyrano-
syl)-(1!4)-6-O-benzyl-2-deoxy-2-phthalimido-b-D-gluco-
pyranoside (37)
˚
A mixture of preactivated molecular sieves 4 A (200 mg),
11 (8.5 mg, 23.0 lmol) and 37 (27.1 mg, 11.5 lmol), and
DTBMP (11.8 mg, 57.5 lmol) in cyclopentyl methyl
ether (CPME) (2 mL) was stirred at room temperature
for 20 min, and MeOTf (6.5 lL, 57.5 lmol) was added.
After being stirred at the same temperature for 10 h, an
additional amount of MeOTf (1.3 lL, 11.5 lmol) was
added, then stirred for 13 h. The reaction was quenched
with Et3N (50 lL) and filtered through Celite. The fil-
trate was diluted with EtOAc, washed with brine, dried
over MgSO4, and evaporated under diminished pressure.
The residue was purified by preparative TLC (1:3 tolu-
ene–EtOAc) and then chromatography with Bio-Beads
˚
A mixture of preactivated molecular sieves 4 A (200 mg)
and compounds 9 (75.5 mg, 37.1 lmol) and 13 (45.2 mg,
74.2 lmol) in CH2Cl2 (3 mL) was stirred at ꢀ78 ꢁC for
20 min. TMSOTf (0.05 M CH2Cl2 soln, 148 lL) was
added and the reaction was stirred at ꢀ50 ꢁC for
1.5 h. It was then quenched with Et3N (100 lL) and fil-
tered through Celite. The filtrate was diluted with
EtOAc, washed with satd aq NaHCO3 and brine, suc-
cessively, dried over MgSO4, and evaporated under
diminished pressure. The residue was purified by chro-
matography on Bio-Beads SX-3 (toluene) and then with
preparative TLC (1:3 hexane–EtOAc) to afford the pro-
tected heptasaccharide. It was then dissolved in CH2Cl2
(3 mL), and Mn(OAc)3Æ2H2O (28.7 mg, 107 lmol) and
DDQ (8.1 mg, 35.8 lmol) were added, successively.
After being stirred for 12 h at room temperature, the
reaction was quenched with satd aq NaHCO3 and di-
luted with EtOAc. The organic layer was washed with
water and brine, successively, dried over MgSO4, and
evaporated under diminished pressure. The residue
was purified by preparative TLC (2:7 toluene–EtOAc)
to afford 62.8 mg (72%, two steps) of 37; 1H NMR
(400 MHz, CDCl3): d 7.82–7.00 (m, 22H, Ar), 5.85
(br, 1H), 5.66 (br, 1H), 5.45 (d, J = 8.8 Hz, 1H), 5.31–
5.25 (m, 3H), 5.17–4.98 (m, 5H), 4.95–4.92 (br, 1H),
4.87 (d, J = 3.2 Hz, 1H), 4.79–4.69 (m, 4H), 4.60 (br,
1H), 4.50 (br, 1H), 4.44–4.22 (m, 9H), 4.14–3.93 (m,
10H), 3.86–3.68 (m, 8H), 3.57–3.42 (m, 5H), 3.24 (br,
2H), 2.26 (s, 3H, CH3CO), 2.10–1.79 (m, 45H, CH3CO);
13C NMR (100 MHz, CDCl3): d 170.4, 170.3, 170.2,
169.8, 169.7, 169.6, 169.5, 169.4, 169.3, 169.2, 169.0,
168.9, 168.7, 167.4, 166.8, 137.8, 136.9, 134.2, 133.7,
131.5, 131.2, 131.1, 127.9, 127.3, 127.1, 127.0, 123.4,
123.1, 98.7, 98.4, 98.2, 97.2, 97.0, 81.7, 74.2, 73.9,
72.7, 72.6, 72.2, 72.1, 70.6, 70.5, 69.7, 69.4, 69.3, 69.2,
68.9, 68.7, 68.6, 68.5, 68.3, 68.2, 67.9, 67.6, 65.6, 65.5,
62.5, 62.3, 62.2, 61.8, 55.9, 54.6, 54.3, 21.1, 21.0, 20.9,
20.8, 20.7, 20.6, 20.5, 20.4; HRESIMS: found m/z
2384.68627 [M+Na]+; calcd for C111H123O54N3Na
2384.68686.
1
SX-3 (toluene) to afford 26.3 mg (85%) of 38; H NMR
(400 MHz, CDCl3): d 7.85–6.90 (m, 27H, Ar), 5.73–
5.63 (m, 2H), 5.47 (d, J 8.5 Hz, 1H), 5.30–5.07 (m,
10H), 5.00 (dd, J 3.4, 10.2 Hz, 1H), 4.93–4.88 (m, 3H),
4.78–4.52 (m, 9H), 4.41–4.15 (m, 12H), 4.09–3.81 (m,
9H), 3.74–3.49 (m, 10H), 3.39–3.34 (m, 2H), 2.24 (s,
3H, CH3CO), 2.10–1.80 (m, 51H, CH3CO), 1.11 (d, J
6.6 Hz, 3H, 6-CH3Fuc); 13C NMR (100 MHz, CDCl3): d
170.5, 170.4, 170.3, 169.9, 169.7, 169.6, 169.5, 169.4,
169.3, 169.2, 169.1, 169.0, 168.9, 167.7, 167.4, 166.9,
138.1, 137.8, 136.8, 134.3, 134.2, 134.1, 133.7, 131.6,
131.3, 131.2, 128.5, 128.2, 127.9, 127.5, 127.4, 127.3,
127.2, 127.1, 123.4, 123.3, 100.4, 98.8, 98.5, 97.9, 97.1,
96.9, 96.7, 96.2, 96.0, 77.6, 75.9, 74.6, 74.2, 73.6, 73.4,
72.8, 72.3, 72.2, 72.1, 72.0, 71.9, 71.6, 71.0, 70.7, 70.6,
70.5, 70.1, 69.3, 69.0, 68.9, 68.8, 68.7, 68.2, 68.0, 65.7,
65.5, 64.6, 62.6, 62.2, 61.9, 56.3, 55.1, 54.4, 21.1, 21.0,
20.9, 20.8, 20.7, 20.6, 16.0; HRESIMS: found m/z
2704.80928 [M+Na]+; calcd for C128H143O60N3Na
2704.81284.
3.27. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
(a-D-mannopyranosyl)-(1!3)-[(a-D-mannopyranosyl)-
(1!6)]-[(b-D-xylopyranosyl)-(1!2)]-(b-D-mannopyranos-
yl)-(1!4)-(2-acetamido-2-deoxy-b-D-glucopyranosyl)-
(1!4)-[(a-L-fucopyranosyl)-(1!6)]-[(a-L-fucopyranosyl)-
(1!3)]-2-acetamido-2-deoxy-D-glucopyranose (1)
Compound 36 (9.4 mg, 3.22 lmol) was treated with eth-
ylenediamine (50 lL) in n-BuOH (1.5 mL) at 85 ꢁC. The
resulting mixture was evaporated under diminished
pressure and the residue was dissolved in pyridine
(1 mL). Subsequently, Ac2O (0.5 mL) was added at

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
691
ice-water temperature, and the reaction was stirred for
6 h at room temperature. The mixture was quenched
with EtOH (2 mL) at ice-cold temperature and evapo-
rated under diminished pressure. The residue was dis-
solved in MeOH (1.5 mL) and MeONa (5 mg) was
added. After being stirred for 12 h, the mixture was
quenched with Amberlyst-15 and filtered. The filtrate
was evaporated under diminished pressure and the resi-
due was purified by Sep-Pakꢂ (C18, 0–50% aq MeOH).
Fractions containing the nonasaccharide were collected
and concentrated. The residue was dissolved in aq
MeOH (20 mL) and 20% Pd(OH)2 (5 mg) was added.
The mixture was stirred under an H2 atmosphere at
room temperature for 12 h and then filtered through
Celite. The filtrate was evaporated under diminished
pressure and filtered through ultrafreeꢂ-MC by centrifu-
gation, then the residue was purified by FPLC (Pharma-
cia, column: Super Peptide 10/300GL, water) to afford
6-CH3Fuc); 13C NMR (125 MHz, D2O, acetone at d
30.89): d 175.4, 175.3, 175.1, 105.8, 101.7, 101.2, 100.4,
100.3, 100.2, 100.0, 99.8, 95.6 · 2, 91.2, 80.3, 79.6, 79.3,
77.8, 77.2, 76.5, 75.9, 75.0, 74.3, 74.0, 73.9, 73.4, 72.8,
72.5, 71.1, 70.6, 70.5, 70.2, 70.1, 69.9, 69.8 · 2, 68.8
67.9, 67.7, 67.5, 67.3, 67.2, 66.1, 65.6, 62.4, 61.6, 61.3,
60.8, 60.7, 56.9, 56.0, 55.7 · 2, 55.6, 54.4, 23.0, 22.5,
16.0; HRESIMS: found m/z 1414.49218 [M+Na]+, calcd
for C53H89O39N3Na 1414.49709.
3.29. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
(a-D-mannopyranosyl)-(1!3)-O-[(a-D-mannopyranosyl)-
(1!6)]-[(b-D-xylopyranosyl)-(1!2)]-(b-D-mannopyranos-
yl)-(1!4)-(2-acetamido-2-deoxy-b-D-glucopyranosyl)-
(1!4)-2-acetamido-2-deoxy-D-glucopyranose (3)
Compound 37 (12.0 mg, 5.08 lmol) was submitted
sequentially to dephthaloylation, acetylation, O-deacet-
ylation, and debenzylation as described for 1 to afford
5.6 mg (88%, four steps) of 3: 1H NMR (400 MHz,
D2O, acetone at d 2.22): d 5.18 (d, J 1.7 Hz, 0.62H, H-
1aGlcN), 5.13 (br s, 1H, H-1aMan), 4.91 (br s, 1H, H-
1aMan), 4.87 (br s, 1H, H-1bMan), 4.69–4.68 (m, 0.38H,
H-1bGlcN), 4.61–4.59 (m, 1H, H-1bGlcN), 4.51 (d, J
8.3 Hz, 1H, H-1bGlcN), 4.43 (d, J 7.5 Hz, 1H, H-1bXyl),
4.25 (d, J 2.7 Hz, 1H, H-2bMan), 4.03 (d, J 3.2 Hz, 1H,
H-2aMan), 3.24 (t, J 11.0 Hz, 1H), 2.07–2.03 (m, 9H,
CH3CO); 13C NMR (100 MHz, D2O, acetone at d
30.89): d 175.0, 174.9, 174.7, 105.6, 101.8, 101.0, 100.2,
100.1, 99.7, 95.3, 90.9, 80.2, 80.1, 79.9, 79.7, 77.6, 77.1,
76.4, 75.9, 75.1, 74.9, 74.2, 73.8, 73.3, 73.0, 72.6, 71.0,
70.6, 70.5, 70.4, 70.0, 69.8, 69.0, 67.7, 67.2, 67.1, 65.9,
65.5, 62.3, 61.5, 61.2, 60.6, 56.7, 55.9, 55.6, 54.2, 23.0,
22.8, 22.5, 20.7; HRESIMS: found m/z 1268.44085
[M+Na]+; calcd for C47H79O35N3Na 1268.43918.
1
4.3 mg (87%) of 1; H NMR (400 MHz, D2O, acetone
at d 2.22): d 5.14 (br s, 1H, H-1aMan), 5.11 (d, J
3.9 Hz, 1H, H-1a1!3Fuc), 5.06 (d, J 3.4 Hz, 0.6H, H-
1aGlcN), 4.92 (d, J 3.9 Hz, 1H, H-1a1!6Fuc), 4.91 (br s,
1H, H-1aMan), 4.85 (br s, 1H, H-1bMan), 4.71–4.66 (m,
1.4H, H-1bGlcN · 2), 4.51 (d, J 8.5 Hz, 1H, H-1bGlcN),
4.45 (d, J 7.6 Hz, 1H, H-1bXyl), 4.25 (br s, 1H, H-2bMan),
3.26 (t, J 11.0 Hz, 1H), 2.05–2.02 (m, 9H, CH3CO), 1.27
(d, J 6.6 Hz, 3H, 6-CHF3 uc), 1.22 (d, J 6.6 Hz, 1.2H,
6-CHF3 uc), 1.20 (d, J 6.6 Hz, 1.8H, 6-CHF3 uc); 13C NMR
(125 MHz, D2O, acetone at d 30.89): d 175.4 · 2,
175.1, 174.9, 105.9, 101.2, 100.7, 100.3, 100.0, 99.8,
99.7 · 2, 81.6, 80.1 · 2, 78.8, 77.8, 77.7, 77.2, 76.5,
75.9, 75.0, 74.3, 74.0, 73.9, 73.4, 73.1, 72.7, 72.5, 71.1,
70.9, 70.6, 70.5, 70.2, 70.1, 69.9 · 2, 69.2, 68.8, 68.4,
67.9, 67.5, 67.3, 67.2, 66.1 · 2, 62.4, 61.6, 61.3, 56.0,
55.7, 54.6, 23.9, 23.0, 22.9, 22.7, 20.7, 16.2, 16.0; HRE-
SIMS: found m/z 1560.55120 [M+Na]+; calcd for
C59H99O43N3Na 1560.55499.
3.30. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
(a-D-mannopyranosyl)-(1!3)-[(a-D-mannopyranosyl)-
(1!6)]-[(b-D-xylopyranosyl)-(1!2)]-(b-D-mannopyranos-
yl)-(1!4)-(2-acetamido-2-deoxy-b-D-glucopyranosyl)-
(1!4)-[(a-L-fucopyranosyl)-(1!3)]-2-acetamido-2-
deoxy-D-glucopyranose (4)
3.28. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
(a-D-mannopyranosyl)-(1!3)-[(a-D-mannopyranosyl)-
(1!6)]-[(b-D-xylopyranosyl)-(1!2)]-(b-D-mannopyrano-
syl)-(1!4)-(2-acetamido-2-deoxy-b-D-glucopyranosyl)-
(1!4)-[(a-L-fucopyranosyl)-(1!6)]-2-acetamido-2-
deoxy-D-glucopyranose (2)
Compound 38 (13.6 mg, 5.07 lmol) was submitted
sequentially to dephthaloylation, acetylation, O-deacetyl-
ation, and debenzylation as described for 1 to afford
4.8 mg (68%, four steps) of 4: 1H NMR (400 MHz,
D2O, acetone at d 2.22): d 5.14 (br s, 1H, H-1aMan),
5.11 (d, J 3.7 Hz, 1H, H-1a1!3Fuc), 5.07 (d, J 3.4 Hz,
0.55H, H-1aGlcN), 4.91 (br s, 1H, H-1aMan), 4.84 (br s,
1H, H-1bMan), 4.69–4.67 (m, 0.45H, H-1bGlcN), 4.56–
4.52 (m, 2H, H-1bGlcN · 2), 4.47 (d, J 7.6 Hz, 1H, H-
1bXyl), 4.25 (d, J 2.7 Hz, 1H, H-2bMan), 4.15 (d, J
2.9 Hz, 1H, H-2aMan), 3.25 (t, J 11.0 Hz, 1H), 2.07–2.02
Compound 34 (13.1 mg, 4.83 lmol) was submitted
sequentially to dephthaloylation, acetylation, O-deacetyl-
ation, and debenzylation as described for compound 1 to
afford 4.9 mg (73%, four steps) of 2; 1H NMR (400 MHz,
D2O, acetone at d 2.22): d 5.17 (d, J 3.4 Hz, 0.67H, H-
1aGlcN), 5.13 (br s, 1H, H-1aMan), 4.91 (br s, 1H, H-
1aMan), 4.90–4.88 (m, 1H, H-1a1!6Fuc), 4.87 (br s, 1H,
H-1bMan), 4.67–4.64 (m, 1.33H, H-1bGlcN · 2), 4.51 (d, J
8.5 Hz, 1H, H-1bGlcN), 4.44 (d, J 7.8 Hz, 1H, H-1bXyl),
4.25 (br s, 1H, H-2bMan), 3.25 (t, J 11.0 Hz, 1H), 2.08–
2.03 (m, 9H, CH3CO), 1.22–1.20 (m, J 6.6 Hz, 3H,
(m, 9H, CH3CO), 1.27 (d, J 6.6 Hz, 3H, 6-CHFuc); 13C
3
NMR (100 MHz, D2O, acetone at d 30.89): d 175.0 · 2,

692
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
174.7, 174.5, 105.6, 101.0, 100.9, 100.2 · 2, 99.6, 98.9,
91.3, 81.3, 79.8, 77.5, 77.0, 76.4, 75.9, 75.1, 74.9, 74.2,
73.8 · 2, 73.3, 73.1, 72.9, 72.6, 71.9, 71.0, 70.5, 70.3,
69.9, 69.8 · 2, 68.3, 67.8, 67.2, 67.1, 65.9, 65.5, 62.3,
61.6, 61.5, 61.2, 57.4, 55.9, 55.7, 54.5, 23.0, 22.9, 22.8,
22.7, 20.7, 16.2; HRESIMS: found m/z 1414.49839
[M+Na]+; calcd for C53H89O39N3Na 1414.49709.
water and brine, successively, dried over MgSO4, and
evaporated under diminished pressure. The residue
was purified by preparative TLC (1:3 toluene–EtOAc)
to afford 24.0 mg (92%) of 40; 1H NMR (400 MHz,
CDCl3): d 7.83–6.95 (m, 22H, Ar), 5.74 (dd, J 9.3,
10.7 Hz, 1H), 5.64 (dd, J 9.3, 10.7 Hz, 1H), 5.39–5.04
(m, 11H), 4.95 (t, J 9.6 Hz, 1H), 4.89 (d, J 3.6 Hz,
1H), 4.80 (d, J 1.5 Hz, 1H), 4.76–4.73 (m, 2H), 4.66–
4.58 (m, 3H), 4.47 (br s, 1H), 4.40–4.01 (m, 13H),
3.88–3.67 (m, 11H), 3.46–3.41 (m, 4H), 3.39–3.34 (m,
1H), 3.26–3.22 (m, 1H), 2.30 (s, 3H, CH3CO), 2.15–
1.84 (m, 45H, CH3CO), 0.98 (d, J 6.6 Hz, 3H,
6-CH3Fuc); 13C NMR (100 MHz, CDCl3): d 170.5,
170.4, 170.2, 170.1, 170.0, 169.9, 169.8, 169.7, 169.6,
169.1, 167.7, 166.7, 138.1, 136.9, 134.5, 134.3, 133.7,
131.6, 131.1, 130.8, 128.4, 128.0, 127.5, 127.4, 127.3,
127.2, 123.9, 123.4, 98.3, 97.8, 97.3, 97.0, 96.9, 96.2,
80.2, 74.1, 74.0, 73.8, 73.3, 72.9, 72.5, 72.2, 72.1, 71.5,
70.6, 70.3, 70.0, 69.5, 69.5, 69.4, 69.3, 69.2, 69.1, 68.9,
68.7, 67.4, 65.6, 65.0, 64.1, 62.3, 62.2, 62.0, 61.8, 55.9,
54.4, 54.3, 29.8, 21.2, 21.0, 20.9, 20.8, 20.7, 20.6, 16.0;
HRESIMS: found m/z 2398.70293 [M+Na]+, calcd for
3.31. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-(2,4-di-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-
fucopyranosyl)-(1!6)]-2-deoxy-3-O-p-methoxybenzyl-2-
phthalimido-b-D-glucopyranoside (39)
Compound 12 (44.6 mg, 24.5 lmol) was glycosylated
with 10 as described for 34 to afford 43.0 mg (70%) of
1
39; H NMR (400 MHz, CDCl3): d 7.87–6.88 (m, 22H,
Ar), 6.83 (d, J 8.5 Hz, 2H, Ar), 6.23 (d, J 8.5 Hz, 2H,
Ar), 5.79 (dd, J 9.3, 10.7 Hz, 1H), 5.64 (dd, J 9.3,
10.7 Hz, 1H), 5.55 (d, J 8.3 Hz, 1H), 5.32–5.21 (m, 6H),
5.17–5.10 (m, 3H), 5.08–4.97 (m, 2H), 4.92–4.90 (m,
1H), 4.77–4.73 (m, 3H), 4.69–4.65 (m, 2H), 4.59–4.56
(m, 2H), 4.44–4.00 (m, 16H), 3.96–3.93 (m, 1H), 3.89–
3.64 (m, 10H), 3.52–3.43 (m, 5H), 3.37–3.26 (m, 2H),
2.27 (s, 3H, CH3CO), 2.27–1.84 (m, 45H, CH3CO), 1.01
(d, J 6.6 Hz, 3H, 6-CHF3 uc); 13C NMR (100 MHz, CDCl3):
d 170.5, 170.4, 170.3, 170.2, 170.0, 169.9, 169.8, 169.7,
169.6, 169.5, 169.4, 169.0, 167.8, 167.1, 158.1, 137.9,
136.7, 134.4, 134.3, 133.1, 131.6, 131.3, 131.0, 130.7,
129.5, 128.3, 128.2, 127.9, 127.8, 127.5, 127.3, 127.2,
123.7, 123.4, 122.9, 122.5, 122.9, 98.3, 97.3, 96.9, 96.7,
96.5, 77.2, 76.4, 76.0, 75.4, 74.2, 74.0, 73.6, 72.5, 72.2,
72.1, 71.9, 71.7, 70.6, 70.0, 69.9, 69.5, 69.3, 69.2, 68.8,
68.6, 67.4, 65.6, 64.8, 64.1, 62.3, 62.0, 61.8, 60.4, 55.6,
55.0, 54.7, 54.3, 21.1, 20.9, 20.8, 20.7, 20.6, 20.5, 20.4,
15.9, 14.3; HRESIMS: found m/z 2518.76413
[M+Na]+; calcd for C120H133O55N3Na 2518.76002.
C
112H125O54N3Na 2398.70251.
3.33. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-(2,4-di-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-
fucopyranosyl)-(1!6)]-[(3,4-di-O-acetyl-2-O-benzyl-a-L-
fucopyranosyl)-(1!3)]-2-deoxy-2-phthalimido-b-D-gluco-
pyranoside (41)
˚
A mixture of preactivated molecular sieves 4 A (200 mg),
compounds 11 (9.50 mg, 25.9 lmol) and 40 (20.5 mg,
8.62 lmol), and DTBMP (8.0 mg, 38.8 mmol) in p-xylene
(1.5 mL) was stirred at room temperature for 30 min.
Subsequently, MeOTf (7.3 lL, 64.7 lmol) was added,
and the mixture was rapidly mixed and frozen by liquid
nitrogen. The mixture was stored in a refrigerator at
4 ꢁC for 84 h and was defrosted at room temperature.
Et3N (100 lL) was added to quench MeOTf and the mix-
ture was filtered through Celite. The filtrate was diluted
with EtOAc, washed with brine, dried over MgSO4,
and evaporated under diminished pressure. The residue
was purified by chromatography with Bio-Beads SX-3
(toluene) and then by preparative TLC (1:3 toluene–
EtOAc) to afford 18.7 mg (80%) of 41; 1H NMR
(400 MHz, CDCl3): d 7.85–6.87 (m, 27H, Ar), 5.79 (dd,
J 8.8, 10.7 Hz, 1H), 5.67–5.62 (m, 2H), 5.29–5.08 (m,
12H), 4.94–4.86 (m, 3H), 4.77–4.53 (m, 8H), 4.43–3.67
(m, 27H), 3.59–3.50 (m, 3H), 3.43–3.36 (m, 2H), 2.25 (s,
3H, CH3CO), 2.10–1.79 (m, 51H, CH3CO), 1.11 (d, J
6.6 Hz, 3H, 6-CH3Fuc), 1.00 (d, J 6.3 Hz, 1H, 6-CHF3 uc);
13C NMR (100 MHz, CDCl3): d 170.7, 170.5, 170.4,
3.32. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-(2,4-di-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-
fucopyranosyl)-(1!6)]-2-deoxy-2-phthalimido-b-D-gluco-
pyranoside (40)
To a stirred soln of 39 (27.5 mg, 11.0 lmol) in CH2Cl2
(1.5 mL) were added Mn(OAc)3Æ2H2O (11.5 mg,
42.9 mmol) and DDQ (3.2 mg, 14.3 mmol), successively.
After being stirred for 13 h at room temperature, the
reaction was quenched with satd aq NaHCO3 and di-
luted with EtOAc. The organic layer was washed with

J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
693
170.3, 169.9, 169.8, 169.6, 169.5, 169.4, 169.1, 169.0,
167.7, 166.9, 137.9, 137.8, 136.6, 134.3, 134.1, 133.7,
131.5, 131.3, 131.2, 131.1, 128.5, 128.2, 128.0, 127.9,
127.6, 127.4, 127.3, 127.2, 123.6, 123.4, 98.5, 98.3, 98.0,
97.1, 97.0, 96.5, 96.4, 96.3, 96.0, 77.2, 75.0, 74.1, 73.6,
72.7, 72.6, 72.2, 72.1, 72.0, 71.9, 71.8, 71.7, 71.6, 71.5,
70.7, 70.4, 70.2, 69.9, 69.4, 69.3, 69.0, 68.9, 68.8, 68.7,
68.2, 65.7, 64.9, 64.8, 64.6, 64.2, 62.6, 62.4, 61.9, 61.8,
56.5, 54.9, 54.3, 21.1, 20.9, 20.8, 20.7, 20.6, 20.5, 20.3,
16.0, 15.9; HRESIMS: found m/z 2718.82648
[M+Na]+; calcd for C129H145O60N3Na 2718.82849.
(34.4 mg, 16.0 lmol), and DTBMP (16.4 mg, 80.0 lmol)
in cyclopentyl methyl ether (CPME) (2.5 mL) was stir-
red at room temperature for 20 min. MeOTf (9.1 lL,
80.0 lmol) was added. After being stirred at the same
temperature for 16 h, MeOTf (0.9 lL, 8.0 lmol) was
added, and the reaction then stirred for 4.5 h. It was
quenched with Et3N (50 lL) and filtered through Celite.
The filtrate was diluted with EtOAc, washed with brine,
dried over MgSO4, and evaporated under diminished
pressure. The residue was purified by chromatography
with Bio-Beads SX-3 (toluene) and then by preparative
TLC (2:5 toluene–EtOAc) to afford 36.3 mg (92%) of
1
3.34. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-(2,4-di-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!4)-2-deoxy-2-phthalimido-b-D-gluco-
pyranoside (42)
43: H NMR (400 MHz, CDCl3): d 7.86–6.91 (m, 27H,
Ar), 5.71 (dd, J 9.0, 10.5 Hz, 1H), 5.65 (dd, J 9.0,
10.7 Hz, 1H), 5.41 (d, J 8.3 Hz, 1H), 5.31–5.09 (m,
10H), 4.93 (d, J 8.3 Hz, 1H), 4.88 (d, J 3.4 Hz, 1H),
4.79 (dd, J 3.4, 10.5 Hz, 1H), 4.74 (d, J 1.2 Hz, 1H),
4.70–4.66 (m, 2H), 4.62–4.59 (m, 2H), 4.55–4.52 (m,
3H), 4.50–4.17 (m, 9H), 4.13–3.71 (m, 13H), 3.66–3.38
(m, 7H), 2.28 (s, 3H, CH3CO), 2.13–1.82 (s, 45H,
Compound 12 (79.1 mg, 43.5 lmol) was submitted
sequentially to glycosylation with 13 and de-p-meth-
oxybenzylation as described for 37. The crude product
was purified by preparative TLC (1:3 toluene–EtOAc)
to afford 75.7 mg (81%, two steps) of 42; 1H NMR
(400 MHz, CDCl3): d 7.82–7.00 (m, 22H, Ar), 5.84
(dd, J 8.8, 10.7 Hz, 1H), 5.64 (dd, J 9.3, 10.7 Hz, 1H),
5.42 (d, J 8.5 Hz, 1H), 5.39–5.28 (m, 2H), 5.23–5.06
(m, 6H), 4.99 (t, J 9.4 Hz, 1H), 4.81 (d, J 1.5 Hz, 1H),
4.75–4.71 (m, 2H), 4.69 (br s, 2H), 4.57–4.01 (m, 17H),
3.87–3.67 (m, 8H), 3.53–3.49 (m, 2H), 3.41–3.37 (br,
1H), 3.23 (br, 2H), 2.30 (s, 3H, CH3CO), 2.16 (s, 3H,
CH3CO), 2.10–1.90 (s, 33H, CH3CO), 1.83 (s, 3H,
CH3CO); 13C NMR (100 MHz, CDCl3): d 170.6,
170.5, 170.4, 170.3, 170.2, 170.0, 169.9, 169.8, 169.7,
169.2, 169.1, 167.6, 166.9, 137.9, 137.0, 134.4, 134.2,
133.8, 131.6, 131.3, 131.1, 130.9, 128.0, 127.9, 127.4,
127.3, 127.1, 123.6, 123.4, 98.6, 98.3, 97.5, 97.1, 97.0,
96.2, 81.7, 77.2, 76.7, 74.6, 74.1, 73.9, 73.7, 73.0, 72.8,
72.2, 72.1, 70.6, 69.7, 69.4, 69.2, 69.1, 68.9, 68.8, 68.7,
68.6, 68.1, 67.5, 65.6, 65.0, 62.4, 62.3, 62.1, 61.8, 55.9,
54.5, 54.3, 21.1, 20.9, 20.8, 20.7, 20.6, 20.5, 20.4; HRE-
SIMS: found m/z 2168.62060 [M+Na]+; calcd for
CH3CO), 1.10 (d, J 6.3 Hz, 3H, 6-CHFuc); 13C NMR
3
(100 MHz, CDCl3): d 170.4, 170.2, 170.1, 169.9, 169.8,
169.7, 169.5, 169.4, 169.3, 169.1, 169.0, 167.8, 166.8,
138.0, 137.8, 136.8, 134.3, 134.1, 133.7, 131.5, 131.2,
131.1, 128.2, 128.1, 127.9, 127.5, 127.4, 127.3, 127.2,
123.5, 123.4, 123.3, 98.4, 98.1, 97.6, 97.0, 96.7, 96.3,
95.9, 76.2, 74.6, 74.5, 74.1, 73.7, 72.8, 72.8, 72.3, 72.1,
72.0, 71.9, 71.8, 71.6, 70.7, 70.4, 70.2, 70.0, 69.9, 69.3,
69.2, 69.1, 68.9, 68.8, 68.7, 68.1, 65.7, 64.9, 64.6, 62.6,
62.3, 62.0, 61.8, 56.3, 55.0, 54.3, 21.1, 21.0, 20.9, 20.8,
20.7, 20.6, 20.5, 16.0; HRESIMS: found m/z
2488.74856 [M+Na]+; calcd for C119H131O54N3Na
2488.74946.
3.36. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
(a-D-mannopyranosyl)-(1!3)-O-[(a-D-mannopyranosyl)-
(1!6)]-(b-D-mannopyranosyl)-(1!4)-(2-acetamido-2-
deoxy-b-D-glucopyranosyl)-(1!4)-[(a-L-fucopyranosyl)-
(1!6)]-2-acetamido-2-deoxy-D-glucopyranose (5)
Compound 39 (12.6 mg, 5.05 lmol) was submitted
sequentially to dephthaloylation, acetylation, O-deacet-
ylation, and hydrogenation as described for 1 to afford
4.8 mg (75%, four steps) of 5; 1H NMR (400 MHz,
D2O, acetone at d 2.22): d 5.17 (d, J 2.7 Hz, 0.67H, H-
1aGlcN), 5.11 (br s, 1H, H-1aMan), 4.91 (br s, 1H, H-
1aMan), 4.89–4.88 (m, 1H, H-1a1!6Fuc), 4.77 (br s, 1H,
H-1bMan), 4.69–4.64 (m, 1.33H, H-1bGlcN · 2), 4.54 (d,
J 8.3 Hz, 1H, H-1bGlcN), 4.24 (br s, 1H, H-2bMan), 4.18
(br s, 1H, H-2aMan), 2.09–2.03 (m, 9H, CH3CO), 1.22–
1.20 (m, 3H, 6-CHF3 uc); 13C NMR (100 MHz, D2O, ace-
tone at d 30.89): d 175.0, 174.9, 174.7, 101.5, 100.9,
100.1, 99.8, 95.5, 91.0, 80.9, 80.3, 79.5, 77.0, 76.4, 74.9,
74.7, 74.1, 73.8, 73.2, 72.6, 72.4, 70.9, 70.7, 70.4, 70.0,
69.9, 69.7, 69.0, 68.8, 67.9, 67.4, 67.3, 66.5, 62.3, 61.5,
61.2, 60.5, 55.9, 55.5, 54.3, 23.0, 22.8, 22.6, 20.7, 16.0;
C
102H111O48N3Na 2168.62347.
3.35. Benzyl (3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-
D-glucopyranosyl)-(1!2)-(3,4,6-tri-O-acetyl-a-D-manno-
pyranosyl)-(1!3)-O-[(2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranosyl)-(1!6)]-(2,4-di-O-acetyl-b-D-mannopyranos-
yl)-(1!4)-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranosyl)-(1!4)-[(3,4-di-O-acetyl-2-O-benzyl-a-L-
fucopyranosyl)-(1!3)]-2-deoxy-2-phthalimido-b-D-gluco-
pyranoside (43)
˚
A
mixture of pre-activated molecular sieves 4 A
(200 mg), compounds 11 (11.8 mg, 32.0 lmol) and 42

694
J. Nakano et al. / Carbohydrate Research 342 (2007) 675–695
HRESIMS: found m/z 1282.45092 [M+Na]+; calcd for
C48H81O35N3Na 1282.45483.
3.39. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
(a-D-mannopyranosyl)-(1!3)-O-[(a-D-mannopyranosyl)-
(1!6)]-(b-D-mannopyranosyl)-(1!4)-(2-acetamido-2-
deoxy-b-D-glucopyranosyl)-(1!4)-[(a-L-fucopyranosyl)-
(1!3)]-2-acetamido-2-deoxy-D-glucopyranose (8)
3.37. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
O-(a-D-mannopyranosyl)-(1!3)-O-[(a-D-mannopyranos-
yl)-(1!6)]-(b-D-mannopyranosyl)-(1!4)-(2-acetamido-
2-deoxy-b-D-glucopyranosyl)-(1!4)-[(a-L-fucopyranos-
yl)-(1!6)]-[(a-L-fucopyranosyl)-(1!3)]-2-acetamido-2-
deoxy-D-glucopyranose (6)
Compound 43 (12.4 mg, 5.03 lmol) was submitted
sequentially to dephthaloylation, acetylation, O-deacet-
ylation, and debenzylation as described for compound
1 to afford 4.4 mg (69%, four steps) of 8: 1H NMR
(400 MHz, D2O, acetone at d 2.22): d 5.11 (br s, 2H,
H-1aMan, H-1a1!3Fuc), 5.07 (d, J 3.4 Hz, 0.55H, H-
1aGlcN), 4.91 (br s, 1H, H-1aMan), 4.74 (br s, 1H, H-
1bMan), 4.71–4.67 (m, 0.45H, H-1bGlcN), 4.57–4.53 (m,
2H, H-1bGlcN · 2), 4.25 (br s, 1H, H-2bMan), 4.18 (d, J
2.0 Hz, 1H, H-2aMan), 2.05–2.02 (m, 9H, CH3CO),
1.27 (d, J 6.8 Hz, 3H, 6-CHF3 uc); 13C NMR (100 MHz,
D2O, acetone at d 30.89): d 175.0 · 2, 174.7, 100.9,
100.2, 100.1, 100.0, 91.3, 81.4, 80.8, 76.9, 76.4, 75.9,
75.1, 74.7, 74.4, 74.3, 74.1, 73.8, 73.2, 72.8, 72.6, 70.9,
70.7, 70.5, 70.4, 69.9, 69.8, 69.0, 68.3, 67.9, 67.3, 66.5,
66.4, 62.3, 61.5, 61.2, 55.9, 55.6, 54.5, 23.0, 22.8, 22.7,
20.7, 16.2; HRESIMS: found m/z 1282.45501
[M+Na]+; calcd for C48H81O35N3Na 1282.45483.
Compound 41 (18.2 mg, 6.75 lmol) was submitted
sequentially to dephthaloylation, acetylation, O-deacet-
ylation, and debenzylation as described for 1 to afford
7.0 mg (74%, four steps) of 6; 1H NMR (400 MHz,
D2O, acetone at d 2.22): d 5.11 (br s, 2H, H-1aMan, H-
1a1!3Fuc), 5.06 (d, J 3.4 Hz, 0.66H, H-1aGlcN), 4.92–
4.91 (m, 2H, H-1aMan, H-1a1!6Fuc), 4.74 (br s, 1H, H-
1bMan), 4.69–4.65 (m, 1.33H, H-1bGlcN · 2), 4.54 (d, J
8.3 Hz, 1H, H-1bGlcN), 4.25 (br s, 1H, H-2bMan), 2.06–
2.02 (m, 9H, CH3CO), 1.27 (d,
J 6.8 Hz, 3H,
6-CHF3 uc), 1.22–1.20 (m, 3H, 6-CHF3 uc); 13C NMR
(125 MHz, D2O, acetone at d 30.89): d 175.4, 174.9,
101.1, 100.8, 100.7, 100.4, 100.3, 100.0, 95.5 · 2, 95.4,
91.6, 81.7 · 2, 81.0, 77.1, 76.5, 75.2, 75.1, 74.8, 74.3,
73.9, 73.4, 73.2, 73.0, 72.7, 72.5, 71.0, 70.9, 70.8, 70.6,
70.2, 70.1, 69.9, 69.8, 69.1, 68.8. 68.4, 68.0, 67.5 · 2,
67.3, 66.6, 62.4, 61.7 · 2, 61.6, 61.3, 56.0, 55.6, 54.6,
23.0, 22.9, 22.7, 20.7, 16.2, 16.0; HRESIMS: found
m/z 1428.50948 [M+Na]+; calcd for C54H91O39N3Na
1428.51274.
Acknowledgments
We thank Ms. A. Takahashi for technical assistance and
Dr. T. Chihara and his staff for elemental analysis and
the high-pressure reaction equipment. Financial support
from the Japan Society for the Promotion of Science
[Grant-in-Aid for Creative Scientific Research No.
17GS0420] is acknowledged.
3.38. (2-Acetamido-2-deoxy-b-D-glucopyranosyl)-(1!2)-
(a-D-mannopyranosyl)-(1!3)-[(a-D-mannopyranosyl)-
(1!6)]-(b-D-mannopyranosyl)-(1!4)-(2-acetamido-2-
deoxy-b-D-glucopyranosyl)-(1!4)-2-acetamido-2-deoxy-
D-glucopyranose (7)
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