Comprehensive synthesis of ER related high-mannose-type sugar chains by convergent strategy

Article abstract of DOI:10.1016/j.tet.2006.06.045

Systematic synthesis of high-mannose-type sugar chains of asparagine-linked glycoproteins is described. To construct the target sugar chains, we employed the convergent route, using three oligosaccharide components, the common hexasaccharide, branched tri

Full text of DOI:10.1016/j.tet.2006.06.045

Tetrahedron 62 (2006) 8262–8277
Comprehensive synthesis of ER related high-mannose-type
sugar chains by convergent strategy
Ichiro Matsuo,a,b Kiichiro Totani,a,b Atsushi Tatamia,b and Yukishige Itoa,b,
*
aRIKEN (The Institute of Physical and Chemical Research), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
bCREST, JST (Japan Science and Technology Agency), Kawaguchi 322-1102, Japan
Received 22 April 2006; revised 14 June 2006; accepted 15 June 2006
Available online 10 July 2006
Abstract—Systematic synthesis of high-mannose-type sugar chains of asparagine-linked glycoproteins is described. To construct the target
sugar chains, we employed the convergent route, using three oligosaccharide components, the common hexasaccharide, branched tri-, tetra-
and pentasaccharides, and mono-, di-, and triglucosyl fragments. Construction of the b-mannoside linkage was performed using p-methoxy-
benzyl-assisted intramolecular aglycon delivery. The hexasaccharide fragment was coupled with the branched mannooligosaccharide donors
such as M5, M4B, M4C, and M3 to give undecasaccharide (M9), decasaccharide (M8B and M8C), and nonasaccharide (M7), respectively.
Incorporation of mono-, di-, and triglucosyl fragments toward them gave tetradecasaccharide (G3M9), tridecasaccharide (G2M9), dodeca-
saccharide (G1M9), undecasaccharide (G1M8B and G1M8C), and decasaccharide (G1M7), respectively.
Ó 2006 Elsevier Ltd. All rights reserved.
1. Introduction
additional mannosidase that may be involved in early pro-
cessing is Man9 mannosidase, which has an ability to trim
M9 to Man7GlcNAc2 (M7) and eventually Man5GlcNAc2
(M5).7
N-Glycosylation of secretory and membrane-bound proteins
is an essential and highly conserved protein modification
of eukaryotes1 (Fig. 1). The key step of protein N-glycosyla-
tion pathway is the en bloc transfer of a triglucosylated
high-mannose-type tetradecasaccharide(Glc3Man9GlcNAc2;
G3M9) from lipid carrier dolichyl pyrophosphate (Dol-PP)
to asparagine (Asn) residue of Asn-X-Ser/Thr sequences of
nascent polypeptide chains. It occurs during their transloca-
tion across the endoplasmic reticulum (ER) membrane.2 This
transformation is catalyzed by a large multisubunit enzyme,
oligosaccharyltransferase (OST) complex. Subsequently,
glucosidase I, an integral membrane protein with a luminally
oriented catalytic domain, removes the terminal a1-2 linked
glucose residue of the triglucosyl sequence. Trimming of
penultimate glucose residues takes place by the action of
glucosidase II to give Glc1Man9GlcNAc2 (G1M9) and then
Man9GlcNAc2 (M9) glycoforms.3,4
Recent investigations have revealed the fundamental roles of
these N-linked high-mannose-type glycans in protein quality
control (folding, transport, and degradation).8–11 A number
of proteins, such as calreticulin (CRT), calnexin (CNX),
ER-Golgi intermediate compartment (ERGIC)-53, vesicular
integral membrane protein (VIP) 36, and mannosidase-like
proteins (MLP, EDEM/Htm1p/Mnl1p) have been revealed
or suggested to recognize these N-linked glycans. CNX
and CRT are ER-resident lectin-chaperones that specifi-
cally recognize monoglucosylated glycans, most typically
G1M9.12,13 They constitute CNX/CRT cycle, together
with UDP-Glc: glycoprotein glucosyltransferase (UGGT).
Intriguingly, the latter enzyme functions as the ‘folding
sensor’, glucosylating prematurely deglucosylated glyco-
proteins having M9, M8 or M7 glycans and regenerate
monoglucosylated structures G1M9w7, in order that they
can engage in multiple rounds of interactions with CNX/
CRT.14 On the other hand, ERGIC-53 and VIP36 are cargo
receptors that mediate glycoprotein transport between ER
and Golgi,15 while MLPs are proposed to capture malfolded
glycoproteins destined for degradation. Although MLPs are
considered to recognize M8B, their lectin activity is yet to be
revealed.
Partial trimming of mannose residues also occurs in the
ER, which is initiated by two distinct enzymes, ER a1-2
mannosidases I and II. The proposed function of the former
enzyme is to remove the terminal sugar of the middle branch
to generate the M8B isomer, which is the proposed ligand
of mannosidase-like protein (MLP).5,6 On the other hand,
mannosidase II preferentially releases the terminal mannose
linked to the a1-6 branch to yield M8C isomer. An
These functions of high-mannose-type glycans in protein
quality control are attracting recent attention. Most of the
* Corresponding author. Tel.: +81 48 467 9430; fax: +81 48 462 4680;
e-mail: yukito@riken.jp
0040–4020/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2006.06.045

I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
8263
Figure 1. (A) Glycoprotein glycan processing in ER. (B) Structure of ER-type N-glycans synthesized in this study.
previous studies were conducted with glycoproteins or
glycan chains derived from biological sources. In order to
gain precise understanding of carbohydrate–protein inter-
plays in protein quality control, access to homogeneous
and structurally defined oligosaccharides is highly impor-
tant. With this respect, approaches based on chemical syn-
thesis are more advantageous, in terms of their flexibility
and ability to provide homogeneous glycans in larger
amount. We wish to describe here the results of our program
toward comprehensive synthesis of ER-related N-glycans.11
Parts of this study have been reported in preliminary forms
(M9 and G1M9,16 M8B and G1M8B,17 G2M9 and G3M9.18
penta-, tri-, and tetramannoside (M5, M3, M4B, and M4C)
fragments, and mono-, di-, and triglucoside donors (G1,
G2, and G3) were chosen. Combination of these fragments
should allow for the construction of all types of high-
mannose-type sugar chains.
2. Results and discussion
2.1. Synthesis of an invariant hexasaccharide
Synthesis of hexasaccharide 11 was carried out as depicted
in Scheme 1. It started with the preparation of trisaccharide
25. Its b-mannoside linkage was constructed using the
2-O-p-methoxybenzyl (PMB) equipped thioglycoside 20
as a donor. Thus, coupling with the 4-O-unprotected fluoride
Considering the diversity of the targets, we adopted a con-
vergent strategy as shown in Figure 2. Thus, fragments cor-
responding to an invariant hexasaccharide 11, branched

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Figure 2. Designed oligosaccharide fragments.
19 was conducted according to our standard protocol; DDQ-
mediated formation of the mixed acetal and subsequent
intramolecular aglycon delivery (IAD) that was promoted
by MeOTf to afford disaccharide 21 as a single stereoisomer
in 84% yield (two steps).19,20 NMR of 21 revealed the pres-
trifluoride (DAST)29 to give corresponding fluorides 12
2
2
[dH 5.66 (d, JH–F¼51.5 Hz)] and 13 [dH 5.59 (d, JH–F
¼
52.0 Hz)] in 89% and 76% yield, respectively. These
were used as M5 (12) and M3 (13) donors for subsequent
transformations.
ence of newly formed b-mannoside linkage [dH 4.37 (J1,2
¼
w0 Hz), dC 99.7 (1JC–H¼159 Hz)].21 After acetylation, it was
condensed with a reducing-end GlcN component 23 through
the activation with Cp2HfCl2/AgOTf22,23 to give 24. Sub-
sequent desilylation proceeded most cleanly under high-
pressure conditions (1 GPa) with HF/pyridine24 to give 25.
Construction of the heterogeneously branched M4C frag-
ment necessitated the discrimination of O-3 and O-6
positions. To that end, compound 35 was regioselectively
protected by treatment with chloroacetic anhydride (CAc2O)
in toluene/dichloromethane (1:1) at 60 ꢀC to give 6-O-CAc
derivative 39.30 The latter was glycosylated with a disaccha-
ride donor 3828 using TfOH as a promoter to give trisaccha-
ride 40 [dC 100.57, 99.17, and 84.27 (C-1)]. Subsequent
removal of the chloroacetyl group provided 41, which was
further glycosylated with 34, giving branched tetramanno-
side 42 in 50% yield [dC 100.52, 99.16, 97.79, and 84.58
(C-1)]. M4B fragment 43 was synthesized from diol 35 as
previously reported.30 These branched oligosaccharides
were converted to fluoride 14 [dH 5.52 (2JH–F¼50.7 Hz)]
and 15 [dH 5.66 (2JH–F¼50.8 Hz)] using NBS–DAST.
The linear mannotriose 31 was synthesized in a stepwise
manner. Coupling of a thiomannoside 26 with chloride 27
was performed under standard conditions (AgOTf/CH2Cl2)
to give mannobiose 28,25,26 which was deacetylated to 29.
As the third mannose residue, the 3-O-TBDPS protected
fluoride 30 was used and trisaccharide 31 was obtained in a
quantitative yield. Three anomeric signals appeared in the
region of a-Man linkage [dH 5.75, 5.54 and 5.21 (s)], while
that of b-Man linkage is typically less than 5 ppm,27 confirm-
ing its stereochemistry. Coupling with 25 was achieved using
MeOTf to afford hexasaccharide 32. Although rigorous
assignment of aMan1-3Man linkage was not made at this
stage, homogeneity of isolated 32 was obvious and we
assumed its stereochemical integrity based on well-estab-
lished intrinsic a-selectivity of mannosyl donor as well
as on our own results. Subsequent removal of the cyclo-
hexylidene group completed the synthesis of the common
hexasaccharide fragment 11.
2.3. Synthesis of oligoglucose fragments
Mono-, di-, and triglucosyl donors were designed as 16, 17,
and 18, respectively (Fig. 2, Scheme 3). Our previous studies
revealed that 4,6-O-benzylidene-protected thioglucoside
was highly suitable for a-selective glucosylation at C-3
position of mannose. The G1 donor 16 was synthesized
from 44. For the synthesis of disaccharide (17) and trisac-
charide (18) fragments, 48 and 52 were chosen as donors,
respectively. We anticipated that the presence of electron-
withdrawing acyloxy groups31 at 3-, 4-, and 6-positions
would be favorable for the stereoselective formation of
a-glucosidic linkages (vide infra).
2.2. Synthesis of oligomannose branch
Syntheses of branched mannooligosaccharide fragments,
M5 (12), M3 (13), M4C (14), and M4B (15) were conducted
as shown in Scheme 2. Preparation of 12 and 13 was straight-
forward, being achieved by double glycosylation of diol
35 with 33/34 to give 36/37, spectral data of which were
identical with previous reported ones.17,28 They were
treated with N-bromosuccinimide (NBS)/diethylaminosulfur
For the preparation of these donors, the 2-OH derivative 49
and hemiacetal 47 were chosen as key intermediates. These
compounds were prepared from glucosyl bromides via
cyclic acetals, as described by Suzuki et al.32,33 Hemiacetal

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8265
Scheme 1. Reagents and conditions: (a) DDQ, CH2Cl2, rt, 3 h; (b) MeOTf, ClCH2CH2Cl, 45 ꢀC, 23 h (84% two steps); (c) Ac2O, DMAP, pyr., 40 ꢀC, 8 h (97%);
(d) Cp2HfCl2, AgOTf, CH2Cl2, ꢁ10 ꢀC, 4 h (78%); (e) 10% HF–pyr., DMF, 1 GPa; (f) AgOTf, ClCH2CH2Cl, ꢁ30 ꢀC to rt, 12 h (83%); (g) NaOMe, MeOH/
THF, rt, 12 h (95%); (h) Cp2HfCl2, AgOTf, CH2Cl2, ꢁ45 to ꢁ20 ꢀC, 1 h (99%); (i) MeOTf, ClCH2CH2Cl, rt, 12 h (82%); (j) TsOH$H2O, CH3CN, rt, 6 h (74%).
47 was converted to fluoride 48 using DASTand used for the
glycosylation with 46 through activation with Cp2HfCl2/
AgOTf. It afforded disaccharide 17 as a single stereoisomer
[dH 5.73 (d, J¼3.6 Hz, H-1a-Glc)] in 63% yield. The G3 frag-
ment 18 was synthesized by the stepwise elongation of glu-
cose residues from the non-reducing end. To begin with,
glycosylation of 49 with 48 using Cp2HfCl2/AgOTf afforded
disaccharide 50 as a single stereoisomer [dH 5.14 (d, J¼
3.6 Hz, H-1a-Glc)] in 92% yield. Removal of the p-methoxy-
benzyl group using DDQ afforded hemiacetal 51 that was
converted to a-imidate 52. Coupling of 52 and 46 through
activation with TMSOTf in toluene provided trisaccharide
and the introduction of glucose fragments were undertaken
as shown in Scheme 4.
Pentamannoside 12 was used as the glycosyl donor to react
with diol 11 using Cp2HfCl2/AgOTf in toluene to provide
undecasaccharide 53 in 87% yield as a single stereoisomer,
which is currently obtainable in multigram quantity (see
Section 4). Similar conditions were applied to 13, 14, and
15, and nonasaccharide 55, as well decasaccharides 57 and
59 were obtained in 77%, 75%, and 84% yield, respectively.
Although we were not able to confirm rigorously their stereo-
chemistry at this stage, contamination of stereoisomer
was not detectable by 400 MHz NMR in each case. Since
selective formation of b-isomer is highly unlikely, we
assumed their structure as depicted. Full confirmation was
made after complete deprotection (vide infra).
1
18 in 45% yield, whose H NMR reveled the presence of
three anomeric protons with a-configuration [dH 5.82
(J¼3.6 Hz), 5.40 (s), 4.93 (J¼3.2 Hz)].
2.4. Systematic synthesis of high-mannose-type glycans
In order to incorporate the pendant glucose residue(s),
removal of the TBDPS group from 53, 55, 57, and 59 was
required. Mindful of the difficulty we previously encountered
With all fragments in hand, the construction of high-man-
nose-type skeletons (undeca-, deca-, and nonasaccharide)

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Scheme 2. Reagents and conditions: (a) 33, AgOTf, ClCH2CH2Cl, ꢁ30 ꢀC to rt (72%); (b) 34, TfOH, ClCH2CH2Cl, ꢁ20 ꢀC (39%); (c) NBS, DAST, CH2Cl2,
ꢁ20 to ꢁ10 ꢀC (89%); (d) NBS, DAST, CH2Cl2, ꢁ30 ꢀC to rt (76%); (e) TfOH, ClCH2CH2Cl, ꢁ20 to ꢁ10 ꢀC (84%); (f) DABCO, EtOH, 50 ꢀC (95%); (g)
TfOH, ClCH2CH2Cl, ꢁ20 to ꢁ10 ꢀC (50%); (h) NBS, DAST, CH2Cl2, ꢁ40 to ꢁ30 ꢀC (87%); (i) NBS, DAST, CH2Cl2, ꢁ30 ꢀC to rt (90%).
in the deprotection of TBDPS groups installed at the O-3-
position of mannose, all of these reactions were conducted
with HF/pyridine under high-pressure reaction conditions.24
For instance, treatment of undecasaccharide 53 in DMF with
10% HF/pyridine at 30 ꢀC under 1 GPa for 24 h cleanly gave
the desired product 54 in 95% yield. In a similar manner, 56,
58, and 60 were obtained from 55, 57, and 59 in 86%, 70%
and 97% yield, respectively.
probes and proteins have been established.11,35–38 Currently,
these glycan chains and derivatives are in extensive use as
molecular probes to clarify various issues related to glyco-
protein quality control system, including specificities of
protein–oligosaccharide interactions and glycan processing
enzymes. These results will be reported in due course.
4. Experimental
4.1. General
As we hoped, incorporation of mono-, di-, and triglucosyl
fragment proceeded selectively to provide desired a-linked
products. Namely, MeOTf-promoted coupling of the unde-
casaccharide 54 with triglucoside 18 gave tetradecasac-
charide 61 in 57% yield, and tridecasaccharide 62 was
obtained in 85% yield under similar conditions, when 17
was used as the donor. Introduction of a glucose residue to
M9, M7, M8C, and M8B type of sugar chain was achieved
using the donor 16 to give 63, 64, 65, and 66 in 93%,
86%, 91%, and 79% yield, respectively.
1H and 13C NMR spectra were measured on JEOL EX-400
spectrometer in CDCl3 and were referenced to Me4Si unless
otherwise mentioned. Silica gel column chromatography
was performed using Silica gel-60 (E Merck). Preparative
thin layer chromatography (PTLC) was developed on E
Merck Silica Gel 60 F254 plates (0.5 mm thickness).
MALDI-TOF MS spectra were recorded in the positive ion
mode on an AXIMA CFR (Shimadzu/KRATOS) equipped
with nitrogen laser with an emission wavelength of
337 nm. High-resolution fast atom bombardment mass
spectrometry was performed on a JEOL IMS-HX-100
mass spectrometer.
Finally, deprotection of these compounds afforded the
ER-type sugar chains in good yield. Figure 3 showed the
1H NMR spectra of these compounds, which were in good
agreement with those reported for closely related com-
pounds.34
4.1.1. Compound 22. Under ice-water cooling, a mixture of
˚
DDQ (3.16 g, 15.9 mmol) and molecular sieves 4 A (15 g) in
3. Conclusion
CH2Cl2 (40 mL) was stirred under Ar. A solution of 20
(9.90 g, 15.3 mmol) and 19 (6.25 g, 12.7 mmol) in CH2Cl2
(60 mL) was added and the mixture was stirred at room tem-
perature for 3 h. It was then quenched with aq solution of
ascorbic acid (0.7%)/citric acid (1.3%)/NaOH (0.9%),
stirred for 5 min, diluted with CH2Cl2/brine, and filtered
In summary, convergent and stereoselective synthetic route
to ER-related N-glycan chains was established. Although
oligosaccharides were prepared as chemically inert n-propyl
glycosides in this study, tactics for the incorporation of

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8267
Scheme 3. Reagents and conditions: (a) PMBCl, Bu4NHSO4, aq NaOH, 45 ꢀC (52%); (b) PivCl, pyr., 0 ꢀC to rt (61%); (c) DAST (92%); (d) Cp2HfCl2, AgOTf,
toluene/ether, ꢁ40 ꢀC (63%); (e) Cp2HfCl2, AgOTf, CH2Cl2, ꢁ10 ꢀC (90%); (f) DDQ, CH2Cl2, H2O, rt (73%); (g) CCl3CN, DBU, CH2Cl2, 0 ꢀC, 1 h (quant.);
(h) TMSOTf, toluene, ꢁ40 ꢀC (45%).
through Celite. The filtrate was washed with aq NaHCO3 and
the organic layers were dried over Na2SO4 and evaporated
in vacuo. The residue was mixed with 2,6-di-tert-butyl-4-
methylpyridine (DTBMP) (10.66 g, 51.91 mmol) and co-
evaporated with toluene three times. The residue was stirred
added a solution of 22 (19.52 g, 19.25 mmol) and 23
(12.23 g, 23.10 mmol) in dry CH2Cl2 (800 mL) at ꢁ10 ꢀC.
The mixture was stirred for 4 h. Insoluble materials were re-
moved by passage through Celite, and the filtrate was then
diluted with CH2Cl2, and washed with brine, aq NaHCO3,
and brine. The organic layer was dried over Na2SO4, and
evaporated in vacuo. The residue was subjected to a silica
gel column chromatography (toluene/EtOAc, 20:1 to 10:1)
to afford compound 24 (22.90 g, 78%). Physical data were
consistent with those reported previously (Ref. 16).
˚
at room temperature with molecular sieves 4 A (30 g) in
ClCH2CH2Cl (200 mL). Then, a solution of 1 M MeOTf in
ClCH2CH2Cl (36.1 mL, 36.1 mmol) was added at 0 ꢀC
and stirred at 45 ꢀC for 23 h. The reaction was quenched
with Et3N, diluted with EtOAc/aq NaHCO3, and filtered
through Celite. The filtrate was washed with aq NaHCO3
and brine, dried over Na2SO4, and evaporated in vacuo.
The residue was subjected to a silica gel column chromato-
graphy (hexane/EtOAc, 5:1) to afford 21 (10.14 g, 82%).
Disaccharide 21 and DMAP (122 mg) were dissolved in-
pyridine (20 mL) and Ac2O (10 mL). The mixture was
stirred at 40 ꢀC for 8 h and evaporated in vacuo. The residue
was diluted with EtOAc, and washed with aq HCl, brine, aq
NaHCO3 and brine. The organic layer was dried over
Na2SO4, and evaporated in vacuo to give 22 (10.27 g,
97%). Physical data were consistent with those reported
previously (Ref. 16).
4.1.3. Compound 25. Compound 24 (16.0 g, 10.5 mmol)
was dissolved in DMF (24 mL) containing 10% HF/
pyridine. The mixture was divided to eight portions and
transferred to 3 mL Teflon reaction vessels. It was com-
pressed to 1.0 GPa and left at 30 ꢀC for 5 h. Combined
mixture was diluted with EtOAc and washed with aq
NaHCO3 and brine, successively. The organic layer was
dried over Na2SO4 and concentrated in vacuo. The combined
mixtures were subjected to a silica gel column chromato-
graphy (hexane/EtOAc¼2:1–3:2) to give compound 25
(11.1 g, 86% yield) as a colorless solid: 1H NMR
(400 MHz, CDCl3) d 7.64–7.00 (m, 28H), 5.50 (m, 1H),
5.27 (d, 1H, J¼2.4 Hz), 5.23 (d, 1H, J¼8.0 Hz), 5.01–4.91
(m, 3H), 4.84 (d, 1H, J¼12.8 Hz), 4.83 (d, 1H,
J¼12.8 Hz), 4.68 (br s, 1H), 4.58 (d, 1H, J¼12.0 Hz),
4.1.2. Compound 24. To a stirred mixture of Cp2HfCl2
(3.73 g, 9.62 mmol), AgOTf (5.19 g, 20.2 mmol), and
˚
molecular sieves 4 A (14.5 g) in dry CH2Cl2 (500 mL) was

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Scheme 4. Reagents and conditions: (a) Cp2HfCl2, AgOTf, toluene, ꢁ30 to ꢁ10 ꢀC; 12 (87%), 13 (77%), 14 (75%), 15 (84%); (b) (1) Ac2O, pyr., 50 ꢀC; (2) 10%
HF–pyr., DMF, 1 Gpa; 54 (95%), 56 (86%), 58 (70%), 60 (97%); (c) MeOTf, CH2CH2, cyclohexane, (1) 61 (54+18; 57%), 62 (54+17; 85%), 63 (54+16; 93%),
64 (56+16; 86%), 65 (58+16; 91%), 66 (60+16; 79%); (d) ethylenediamine, n-BuOH; (e) Ac2O, pyr.; (f) NaOMe, MeOH; (g) Pd(OH)2/C, aq AcOH, H2, 1 (54%),
2 (84%), 3 (47%), 4 (87%), 5 (56%), 6 (65%), 7 (54%), 8 (84%), 9 (66%), 10 (58%).
Figure 3. 1H NMR spectra of high-mannose-type oligosaccharides.

I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
8269
4.51–4.36 (m, 5H), 4.23–4.09 (m, 7H), 3.87 (m, 1H), 3.73–
3.69 (m, 2H), 3.61–3.38 (m, 6H), 3.29 (m, 1H), 3.18 (m, 1H),
2.98 (m, 1H), 2.18 (s, 3H); 13C NMR (100 MHz, CDCl3)
d 170.16, 167.22, 138.52, 138.42, 138.08, 137.69, 137.61,
133.73, 133.55, 133.28, 131.52, 131.25, 128.82, 128.34,
128.06, 128.02, 127.92, 127.84, 127.70, 127.61, 127.58,
127.38, 127.29, 127.13, 126.86, 126.64, 125.10, 123.41,
122.19, 116.93, 99.87, 99.09, 97.02, 98.87, 78.92, 76.82,
75.71, 74.47, 74.44, 73.16, 72.93, 72.71, 71.35, 71.24,
70.38, 70.15, 69.43, 68.15, 67.82, 67.60, 61.02, 60.35,
56.54, 55.66, 41.99, 37.92, 27.99, 27.07, 25.62, 25.07,
22.82, 22.58, 21.53, 21.16, 21.11, 14.30; HRMS (FAB)
m/z calcd for C73H76N2O19Na 1307.4940 (M+Na)+, found
1307.4940.
4.1.8. Compound 11. To a stirred solution of compound 32
(4.29 g, 1.74 mmol) in dry CH3CN was added p-toluenesul-
fonic acid monohydrate (0.83 g, 4.4 mmol) and stirred for
6 h at room temperature. The reaction was quenched with
TEA (0.1 mL) and concentrated in vacuo. The residue was
subjected to a silica gel column chromatography (toluene/
EtOAc, 10:1) to afford the compound 11 (3.54 g, 74%).
Physical data were consistent with those reported previously
(Ref. 16).
4.1.9. Compound 36. A mixture of AgOTf (14.8 g,
˚
57.6 mmol), molecular sieves 4 A (100 g), and compound
35 (5.64 g, 12.8 mmol) in dry ClCH2CH2Cl (100 mL) was
stirred at 0 ꢀC for 30 min, then cooled at ꢁ30 ꢀC. The mix-
ture was added a solution of 33 (20.2 g, 28.9 mmol) in dry
ClCH2CH2Cl (100 mL) dropwise over 15 min. The reaction
mixture was stirred at ꢁ30 ꢀC for 1 h and at ambient tem-
perature (12 h). It was then quenched with TEA (1 mL)
and processed as described for 28. The residue was subjected
to silica gel column chromatography (hexane/EtOAc, 1:1
to 1:3) to afford 36 (21.0 g, 72%). Physical data were consis-
tent with those reported previously (Ref. 28).
4.1.4. Compound 28. A mixture of AgOTf (12.7 g,
˚
49.6 mmol) and molecular sieves 4 A (30 g) in dry toluene
(105 mL) was stirred at ꢁ40 ꢀC for 30 min. A solution of
compounds 27 (10.8 g, 21.3 mmol) and 26 (8.5 g,
18 mmol) in dry ClCH2CH2Cl (159 mL) was added drop-
wise over 40 min and the mixture was stirred at ꢁ30 ꢀC
for 1 h and at ambient temperature for 12 h. The reaction
was quenched with TEA (20 mL). The reaction mixture
was diluted with EtOAc and filtered through Celite. The fil-
trate was washed with aq NaHCO3 and brine, successively.
The organic layer was dried over Na2SO4 and concentrated
in vacuo. The residue was subjected to a silica gel column
chromatography (hexane/EtOAc, 10:1 to 1:1) to afford 28
(14.1 g, 83%). Physical data were consistent with those
reported previously (Ref. 16).
4.1.10. Compound 12. A mixture of compound 36 (6.00 g,
3.56 mmol) and NBS (948 mg, 5.33 mmol) in CH2Cl2
(100 mL) was added DAST (1.41 mL, 10.6 mmol) at
ꢁ30 ꢀC. The mixture was stirred at ꢁ30 ꢀC for 1 h and at
ambient temperature (3 h). The reaction mixture was pro-
cessed as described for 28. The residue was subjected to a
silica gel column chromatography (hexane/EtOAc, 3:2) to
afford 12 (5.05 g, 89%) as a colorless solid: 1H NMR
(400 MHz, CDCl3) d 7.38–7.29 (m, 10H), 5.66 (d, 1H,
J¼51.5 Hz), 5.40–5.21 (m, 10H), 5.07 (br s, 1H), 4.89 (d,
1H, J¼12 Hz), 4.81–4.64 (m, 4H), 4.20–3.71 (m, 19H),
2.15 (s, 3H), 2.13 (s, 3H), 2.11 (s, 3H), 2.09 (s, 3H), 2.087
(s, 3H), 2.07 (s, 3H), 2.05 (s, 9H), 2.04 (s, 3H), 2.03 (s,
6H), 2.00 (s, 6H); 13C NMR (100 MHz, CDCl3) d 170.73,
170.55, 170.39, 170.31, 170.13, 169.61, 169.57, 169.54,
169.34, 169.30, 169.20, 169.10, 137.48, 137.08, 128.51,
128.40, 127.84, 127.73, 127.55, 127.15, 105.51, 100.08,
99.25, 99.14, 99.08, 78.38, 77.78, 77.49, 75.79, 75.45,
74.91, 74.25, 73.44, 72.41, 70.10, 69.91, 69.83, 69.67,
69.53, 69.27, 69.10, 68.55, 68.40, 66.25, 66.19, 66.11,
65.90, 62.45, 62.28, 62.13, 61.89, 20.86, 20.81, 20.68,
20.52; MALDI-TOF mass m/z calcd for C72H91FO39Na
1621.5 (M+Na)+, found 1622.2.
4.1.5. Compound 29. To a stirred solution of 28 (14.1 g,
14.8 mmol) in THF/MeOH (1:1, 148 mL) was added 28%
NaOMe/MeOH (1.0 mL) at room temperature. The mixture
was stirred for 12 h, neutralized with Amberlyst 15 (H+)
resin, and evaporated in vacuo. The residue was subjected
to a silica gel column chromatography (hexane/EtOAc, 5:1
to 1:1) to give 29 (12.9 g, 95%). Physical data were consis-
tent with those reported previously (Ref. 16).
4.1.6. Compound 31. To a stirred mixture of Cp2HfCl2
(5.71 g, 15.0 mmol), AgOTf (7.74 g, 30.1 mmol), and
˚
molecular sieves 4 A (5 g) in dry CH2Cl2 (200 mL) was
added a solution of 30 (8.04 g, 11.6 mmol) and 29 (9.17 g,
10.0 mmol) in dry CH2Cl2 (50 mL) at ꢁ45 ꢀC. The mixture
was gradually warmed up to ꢁ20 ꢀC and stirred for 1 h. The
reaction was quenched with TEA (20 mL) and processed as
described for 28. The residue was subjected to a silica gel
column chromatography (hexane/EtOAc, 10:1 to 5:1) to
afford the compound 31 (15.9 g, 99%). Physical data were
consistent with those reported previously (Ref. 16).
4.1.11. Compound 37. A mixture of 35 (1.11 g, 2.45 mmol),
TfOH (20 mL), and molecular sieves AW 300 (6 g) in dry
ClCH2CH2Cl (20 mL) was stirred at ꢁ20 ꢀC for 30 min.
The donor 33 (2.90 g, 5.89 mmol) in dry ClCH2CH2Cl
(5 mL) was added dropwise over 10 min. The reaction was
quenched with aq NaHCO3 and processed as described for
28. The residue was subjected to a silica gel column chro-
matography (hexane/EtOAc, 5:1 to 1:1) to afford 37
(1.01 g, 39%). Physical data were consistent with those
reported previously (Ref. 28).
4.1.7. Compound 32. A mixture of 25 (2.73 g, 2.12 mmol),
˚
31 (5.05 g, 3.19 mmol), and molecular sieves 4 A (15 g) in
dry CH2Cl2 (200 mL) was stirred at ꢁ40 ꢀC for 30 min, to
which was added 1 M MeOTf (31.8 mL, 31.8 mmol) in
ClCH2CH2Cl. The mixture was stirred at ꢁ40 ꢀC for
30 min and at ambient temperature (12 h). The reaction
was quenched with TEA (10 mL) and processed as described
for 28. The residue was subjected to a silica gel column chro-
matography (toluene/EtOAc, 30:1) to afford 32 (4.92 g,
82%). Physical data were consistent with those reported
previously (Ref. 16).
4.1.12. Compound 13. A mixture of compound 37 (229 mg,
0.206 mmol) and NBS (73.4 mg, 0.412 mmol) in CH2Cl2
(2 mL) was added DAST (82 mL, 0.62 mmol) at ꢁ40 ꢀC.
The reaction mixture was stirred at ꢁ30 ꢀC for 1 h and
then at ambient temperature. After 12 h, MeOH (0.1 mL)

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I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
was added and the mixture was diluted with EtOAc, washed
successively with aq NaHCO3 and brine. The organic layer
was dried over Na2SO4 and concentrated in vacuo. The
residue was subjected to a silica gel column chromatography
(toluene/EtOAc, 5:1 to 2:1) to afford compound 13 (162 mg,
76%) as a colorless solid: 1H NMR (400 MHz, CDCl3)
d 7.44–7.24 (m, 10H), 5.59 (d, 1H, J¼52.0 Hz), 5.40–5.17
(m, 9H), 5.15 (br s, 1H), 4.90 (d, 1H, J¼2.0 Hz), 4.87 (d,
1H, J¼11.6 Hz), 4.80 (d, 1H, J¼12.4 Hz), 4.71 (d, 1H,
J¼12.4 Hz), 4.64 (d, 1H, J¼11.6 Hz), 4.21 (dd, 1H,
J¼4.8, 12.0 Hz), 4.14–3.72 (m, 16H), 2.14 (s, 3H), 2.07 (s,
3H), 2.06 (s, 3H), 2.05 (s, 3H), 2.04 (s, 3H), 2.02 (s, 3H),
2.01 (s, 3H), 1.97 (s, 3H); 13C NMR (100 MHz, CDCl3)
d 170.36, 170.26, 170.16, 169.47, 169.43, 169.37, 169.32,
169.27, 169.24, 137.18, 137.09, 128.48, 128.41, 128.34,
128.32, 128.22, 127.85, 127.72, 127.66, 127.57, 127.56,
105.26, 99.05, 97.94, 75.05, 73.90, 73.85, 72.60, 69.37,
69.31, 68.92, 68.84, 68.71, 68.47, 66.38, 66.13, 66.06,
65.98, 62.46, 62.23, 20.92, 20.75, 20.69, 20.59; MALDI-
TOF mass m/z calcd for C48H59FO23Na 1045.3 (M+Na)+,
found 1045.4.
mixture was processed as described for 28. The residue
was subjected to a silica gel column chromatography
(hexane/EtOAc, 5:1 to 1:2) to afford 42 (249 mg 50%) as
a colorless amorphous: 1H NMR (400 MHz, CDCl3)
d 7.45–7.27 (m, 15H), 5.62 (d, 1H, J¼1.2 Hz), 5.36–5.21
(m, 9H), 4.89 (d, 1H, J¼10.8 Hz), 4.81 (d, 1H, J¼
12.0 Hz), 4.70 (d, 1H, J¼2.0 Hz), 4.64 (d, 1H, J¼
11.6 Hz), 4.49 (d, 1H, J¼12.4 Hz), 4.23–3.80 (m, 17H),
2.15 (s, 3H), 2.14 (s, 3H), 2.12 (s, 6H), 2.10 (s, 3H), 2.04
(s, 3H), 2.04 (s, 3H), 2.04 (s, 3H), 2.02 (s, 3H), 2.01 (s,
3H), 1.98 (s, 3H); 13C NMR (100 MHz, CDCl3) d 170.49,
170.31, 169.98, 169.51, 169.45, 169.35, 169.33, 169.10,
168.95, 137.66, 137.14, 133.64, 131.35, 129.08, 128.43,
128.33, 127.60, 127.55, 127.42, 127.39, 100.52, 99.16,
97.79, 84.58, 78.74, 77.82, 75.23, 74.62, 72.22, 71.00,
70.04, 69.63, 69.38, 69.29, 68.99, 68.95, 68.37, 66.70,
66.22, 66.08, 65.92, 62.58, 62.29, 62.15, 20.99, 20.85,
20.83, 20.79, 20.77, 20.75; MALDI-TOF mass m/z calcd
for C66H80O31SNa 1423.4 [M+Na]+, found 1422.7.
4.1.16. Compound 14. Compound 42 (180 mg, 0.128 mmol)
was treated with NBS (68 mg, 0.39 mmol) and DAST
(51 mL, 0.39 mmol) as described for 13 and purified by silica
gel column chromatography (toluene/EtOAc, 5:1 to 2:3) to
afford compound 14 (146 mg, 87%) as a colorless solid:
1H NMR (400 MHz, CDCl3) d 7.28–7.05 (m, 10H), 5.52
(d, 1H, J¼50.7 Hz), 5.27–5.10 (m, 7H), 4.80–4.53 (m,
5H), 4.17–3.57 (m, 14H), 2.04 (s, 3H), 2.03 (s, 3H), 1.99
(s, 3H), 1.97 (s, 3H), 1.95 (s, 3H), 1.94 (s, 6H), 1.93 (s,
6H), 1.92 (s, 3H), 1.92 (s, 3H); 13C NMR (100 MHz,
CDCl3) d 170.52, 170.40, 170.29, 170.05, 169.58, 169.51,
169.49, 169.22, 169.06, 137.49, 137.11, 128.84, 128.49,
128.41, 128.36, 128.03, 127.86, 127.79, 127.68, 127.63,
127.60, 127.51, 127.40, 105.17, 100.26, 99.14, 98.02,
78.71, 77.81, 75.01, 73.97, 73.44, 72.44, 69.84, 69.53,
69.34, 69.24, 69.03, 68.82, 68.43, 68.34, 66.23, 66.12,
66.04, 65.90, 62.46, 62.16, 20.83, 20.71, 20.66, 20.63,
20.50; MALDI-TOF mass m/z calcd for C60H75FO31Na
1333.4 (M+Na)+, found 1333.0.
4.1.13. Compound 40. A mixture of compound 39 (0.497 g,
0.939 mmol), TfOH (10 mL), and molecular sieves AW 300
(4 g) in dry ClCH2CH2Cl (10 mL) was stirred at ꢁ20 ꢀC for
30 min. A solution of the glycosyl donor 38 (0.90 g,
1.2 mmol) in dry ClCH2CH2Cl (10 mL) was added dropwise
over 10 min and stirred at ꢁ10 ꢀC for 8 h. The reaction was
quenched with aq NaHCO3 and processed as described for
28. The residue was subjected to a silica gel column chro-
matography (toluene/EtOAc, 5:1 to 1.5:1) to afford com-
pound 40 (0.89 g, 84%) as a colorless amorphous: 1H
NMR (400 MHz, CDCl3) d 7.44–7.27 (m, 15H), 5.66 (d,
1H, H-1), 5.38–5.23 (m, 6H), 4.85 (d, 1H, J¼11.6 Hz),
4.77 (d, 1H, J¼11.6 Hz), 4.71 (d, 1H, J¼1.6 Hz), 4.60 (d,
1H, J¼11.6 Hz), 4.50 (d, 1H, J¼11.6 Hz), 4.37–3.93 (m,
16H), 2.15 (s, 3H), 2.14 (s, 3H), 2.11 (s, 3H), 2.10 (s, 3H),
2.05 (s, 3H), 2.04 (s, 3H), 2.01 (s, 3H); 13C NMR
(100 MHz, CDCl3) d 170.42, 170.24, 170.05, 169.48,
169.39, 169.08, 168.89, 166.56, 166.56, 137.31, 136.95,
133.23, 131.30, 128.99, 128.39, 128.34, 127.99, 127.71,
127.65, 127.61, 127.34, 100.57, 99.17, 84.27, 81.01, 78.72,
77.75, 75.24, 74.16, 71.05, 70.63, 69.98, 69.60, 69.29,
69.05, 68.30, 66.19, 65.86, 64.53, 62.60, 62.31, 40.63,
20.93, 20.82, 20.75, 20.73, 20.71; MALDI-TOF mass m/z
calcd for C54H63ClO23SNa 1169.3 (M+Na)+, found 1168.5.
4.1.17. Compound 15. Compound 43 (537 mg, 0.382 mmol)
was treated with NBS (135 mg, 0.759 mmol) and DAST
(0.10 mL, 0.75 mmol) as described for 13 and purified by
silica gel column chromatography (toluene/EtOAc, 5:1 to
2:1) to afford compound 15 (452 mg, 90%) as a colorless
1
solid: H NMR (400 MHz, CDCl3) d 7.38–7.29 (m, 10H),
5.66 (d, 1H, J¼50.8 Hz), 5.41–5.10 (m, 23H), 4.89–4.64
(m, 5H), 4.21–3.75 (m, 30H), 2.11 (s, 3H), 2.10 (s, 3H),
2.07 (s, 3H), 2.06 (s, 6H), 2.05 (s, 3H), 2.04 (s, 3H), 2.03
(s, 3H), 2.02 (s, 3H), 2.01 (s, 3H), 2.00 (s, 3H); 13C NMR
(100 MHz, CDCl3) d 170.81, 170.47, 170.43, 170.09,
169.64, 169.60, 169.49, 169.30, 169.28, 137.32, 137.18,
136.92, 128.89, 128.61, 128.53, 128.34, 128.08, 127.86,
127.78, 127.67, 127.45, 125.16, 105.51, 100.08, 99.25,
99.14, 99.08, 77.41, 75.94, 75.42, 75.10, 74.24, 73.86,
72.61, 71.89, 69.95, 69.71, 69.31, 69.10, 69.02, 68.73,
68.61, 68.43, 66.29, 66.16, 65.98, 62.50, 62.34, 61.99,
20.85, 20.71, 20.66, 20.56; MALDI-TOF mass m/z calcd
for C60H75FO31Na 1333.4 (M+Na)+, found 1332.7.
4.1.14. Compound 41. Trisaccharide 40 (453 mg,
0.423 mmol) was dissolved in EtOH (20 mL) and treated
with DABCO (300 mg) at 50 ꢀC for 2 h. The mixture was
neutralized with Amberlist 15 E [H+]. Insoluble materials
were removed by filtration and the filtrate was concentrated
in vacuo. The residue was subjected to a silica gel column
chromatography (toluene/EtOAc, 3:1 to 1:1) to give 41
(430 mg, 95%). Physical data were consistent with those
reported previously (Ref. 28).
4.1.15. Compound 42. A mixture of compound 41 (377 mg,
0.352 mmol), TfOH (15 mL), and molecular sieves AW 300
(4 g) in dry ClCH2CH2Cl (10 mL) was stirred at ꢁ20 ꢀC
for 10 min. A solution of glycosyl donor 34 (472 mg,
0.958 mmol) in dry ClCH2CH2Cl (10 mL) was added drop-
wise over 10 min and stirred at ꢁ10 ꢀC for 1 h. Resulting
4.1.18. Compound 16. To a stirred solution of compound 44
(73.0 mg, 0.245 mmol), p-methoxybenzyl chloride (41 mL),
Bu4NHSO4 (40 mg) in CH2Cl2 (4 mL) was added 5% aq

I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
8271
NaOH (0.5 mL). The mixture was stirred at 45 ꢀC for 12 h.
The reaction mixture was diluted with CHCl3 and washed
with brine. The organic layer was dried over Na2SO4 and
evaporated in vacuo. The residue was subjected to a PTLC
(toluene/EtOAc, 8:1) to afford compound 45 (53.2 mg,
52%) and its regioisomer (33.0 mg, 32%). Compound 45
(327 mg, 0.782 mmol) was dissolved in pyridine (3 mL)
and added PivCl (0.57 mL) at 0 ꢀC. The mixture was stirred
at room temperature for 12 h. The reaction was added MeOH
(1 mL) and evaporated in vacuo, The residue was diluted
with EtOAc and washed with aq CuSO4, brine, satd aq
NaHCO3, and brine. The organic layer was dried over
NaSO4 and evaporated in vacuo. The residue was crystal-
lized from 2-propanol to give 16 (238 mg, 61%) as a color-
less needles: mp 124–125 ꢀC; 1H NMR (400 MHz, CDCl3)
d 7.42–6.85 (m, 9H), 5.58 (s, 1H), 5.41 (t, 1H, J¼9.2 Hz),
4.52 (d, 1H, J¼9.2 Hz), 4.58–4.53 (m, 2H), 4.38 (dd, 1H,
J¼4.8, 10.4 Hz), 3.97 (s, 3H), 3.63 (t, 1H, J¼9.6 Hz),
3.55–3.51 (m, 2H), 2.26 (s, 3H), 1.22 (s, 9H); 13C NMR
(100 MHz, CDCl3) d 177.01, 159.26, 136.86, 129.58,
129.38, 128.79, 128.07, 125.80, 113.75, 100.97, 86.31,
79.15, 79.14, 74.75, 74.33, 70.19, 68.63, 55.31, 27.23,
13.46; Anal. Calcd for C27H34O7S: C, 64.52; H, 6.92; S,
6.38. Found: C, 64.26; H, 6.77; S, 6.20.
78.62, 75.54, 75.17, 71.33, 70.57, 69.87, 69.02, 68.72,
67.56, 62.05, 21.42, 12.85; MALDI-TOF mass m/z calcd
for C55H52O13SNa 975.3 (M+Na)+, found 975.7.
4.1.21. Compound 50. A mixture of Cp2HfCl2 (80.9 mg,
0.213 mmol), AgOTf (110 mg, 0.426 mmol), and molecular
˚
sieves 4 A (1.4 g) in dry CH2Cl2 (2 mL) was stirred at
ꢁ10 ꢀC. A solution of glycosyl donor 48 (83.5 mg,
0.142 mmol) and glycosyl acceptor 49 (52 mg 0.070 mmol)
in dry CH2Cl2 (2 mL) was added. The mixture was stirred
at ꢁ10 ꢀC for 7 h and processed as described for 28. The resi-
due was subjected to a column of Bio-Beads SX-4 (toluene)
to afford compound 50 (82 mg, 90%) as a colorless amor-
1
phous: H NMR (400 MHz, CDCl3) d 8.01–7.12 (m, 36H),
6.03 (t, 1H, J¼9.6 Hz), 5.82 (t, 1H, J¼9.6 Hz), 5.41 (t, 1H,
J¼9.6 Hz), 5.29 (t, 1H, J¼9.6 Hz), 5.14 (d, 1H, J¼3.6 Hz),
4.93 (d, 1H, J¼3.2 Hz), 4.66 (d, 1H, J¼11.6 Hz), 4.51 (d,
1H, J¼11.6 Hz), 4.48–4.40 (m, 2H), 4.35–4.28 (m, 2H),
4.22 (m, 1H), 4.05–3.98 (m, 3H), 3.82 (s, 3H), 3.77 (s, 3H),
3.73 (s, 3H), 3.55 (s, 3H); 13C NMR (100 MHz, CDCl3)
d 165.85, 165.76, 165.18, 165.07, 165.02, 164.97, 163.48,
163.29, 163.18, 159.31, 137.31, 133.01, 132.92, 132.82,
131.93, 131.66, 130.01, 129.69, 129.58, 129.57, 128.73,
128.59, 128.30, 128.16, 128.13, 127.78, 122.12, 121.68,
121.25, 113.83, 113.55, 96.91, 94.98, 72.56, 71.78, 71.23,
69.83, 69.72, 69.36, 68.11, 67.99, 63.12, 62.64, 55.40,
55.21, 55.15; MALDI-TOF mass m/z calcd for
C72H66O21Na 1289.4, found 1290.3.
4.1.19. Compound 48. To a solution of compound 47
(378 mg, 0.649 mmol) in CH2Cl2 (10 mL) was added
DAST (171 mL, 1.30 mmol) at ꢁ40 ꢀC for 30 min. MeOH
(0.5 mL) was added to the reaction mixture, which was
diluted with EtOAc and washed successively with aq
NaHCO3, and brine. The organic layer was dried over
Na2SO4, concentrated and subjected to a silica gel column
chromatography (toluene/EtOAc, 5:1) to afford 48
4.1.22. Compound 52. To a solution of 50 (264 mg,
0.209 mmol) in CH2Cl2 (5 mL) was added DDQ (273 mg,
1.20 mmol), followed by H2O (1 mL). The mixture was
stirred at room temperature for 12 h, diluted with EtOAc,
quenched with ascorbate buffer. The organic layer was
washed with brine, dried (Na2SO4), and concentrated
in vacuo. The residue was subjected to a silica gel column
chromatography (hexane/EtOAc, 5:1 to 2:3) to give hemi-
acetal 51 (175 mg, 73%): MALDI-TOF mass m/z calcd for
C64H58O20Na 1169.3 (M+Na)+, found 1169.8. Hemiacetal
51 (157 mg, 0.137 mmol) was dissolved in CCl3CN
(1 mL) and CH2Cl2 (1 mL) and added DBU (10 mL) at
0 ꢀC. After stirring for 1 h, the mixture was subjected to a
silica gel column chromatography (toluene/EtOAc, 10:1 to
3:1 in 0.1% TEA) to give 52 (177 mg, quant.) as a slightly
1
(348 mg, 92%) as a colorless solid: H NMR (400 MHz,
CDCl3) d 7.94–7.06 (m, 20H), 5.64–5.53 (m, 2H), 5.44
(dd, 1H, J¼6.0, 46.0 Hz), 4.74 (d, 1H, J¼12.0 Hz), 4.62–
4.53 (m, 3H), 4.38 (dd, 1H, J¼5.2, 12.4 Hz), 4.16 (m, 1H),
3.73 (m, 1H); 13C NMR (100 MHz, CDCl3) d 109.12
(d, J¼220.1 Hz), 77.72 (d, J¼25.1 Hz), 73.62, 72.81,
72.02, 68.67, 62.89; MALDI-TOF mass m/z calcd for
C34H29FO8Na 607.6 (M+Na)+, found 607.7.
4.1.20. Compound 17. A mixture of Cp2HfCl2 (187 mg,
0.493 mmol), AgOTf (273 mg, 1.06 mmol), and molecular
˚
sieves 4 A (1.4 g) was added a solution of compounds 48
1
yellow amorphous: H NMR (400 MHz, CDCl3) d 8.62 (s,
(160 mg, 0.274 mmol) and 46 (107 mg, 0.274 mmol) in
ether/toluene (2:1, 30 mL) at ꢁ40 ꢀC. The mixture was
stirred at ꢁ40 ꢀC for 9 h. The reaction was quenched with
TEA and processed as described for 28. The residue was
subjected to a column of Bio-Beads SX-4 (toluene) to afford
17 (165 mg, 63%) as a colorless amorphous: 1H NMR
(400 MHz, CDCl3) d 8.02–6.69 (m, 30H), 6.01 (t, 1H,
J¼9.6 Hz), 5.73 (d, 1H, J¼3.6 Hz), 5.47 (s, 1H), 5.41 (t,
1H, J¼9.9 Hz), 5.20 (d, 1H, J¼10.4 Hz), 4.93 (d, 1H,
J¼10.4 Hz), 4.51 (d, 1H, J¼9.6 Hz), 4.36 (dd, 1H, J¼4.8,
10.4 Hz), 4.25–4.11 (m, 3H), 3.89 (t, 1H, J¼9.6 Hz), 3.83–
3.75 (m, 2H), 3.69 (t, 1H, J¼9.2 Hz), 3.60 (dd, 1H, J¼3.6,
10.0 Hz), 3.54 (m, 1H), 2.29 (s, 3H); 13C NMR (100 MHz,
CDCl3) d 165.83, 165.60, 165.06, 137.11, 136.77, 136.73,
133.08, 132.86, 132.69, 129.76, 129.71, 129.62, 129.58,
129.42, 129.38, 128.91, 128.85, 128.35, 128.19, 128.10,
128.05, 128.02, 128.01, 127.97, 127.68, 127.52, 127.41,
127.19, 126.33, 125.13, 102.21, 95.56, 86.09, 81.97,
1H), 8.02–6.63 (m, 33H), 6.13 (t, 1H, J¼10.0 Hz), 5.77 (t,
1H, J¼9.8 Hz), 5.57 (t, 1H, J¼10.1 Hz), 5.34 (t, 1H,
J¼10.0 Hz), 5.08 (d, 1H, J¼3.4 Hz), 4.35 (dd, 1H, J¼5.3,
12.4 Hz), 4.25 (m, 2H), 4.11 (m, 1H), 3.94 (m, 1H), 3.83
(s, 3H), 3.79 (s, 3H), 3.69 (dd, 1H, J¼3.4, 10.0 Hz), 3.57
(s, 3H).
4.1.23. Compound 18.
A mixture of 52 (65 mg,
0.057 mmol), 46 (16 mg, 0.042 mmol), and molecular sieves
AW 300 (700 mg) in dry toluene (3 mL) was stirred at
ꢁ40 ꢀC for 30 min, to which was added TMSOTf (5 mL).
The mixture was stirred at ꢁ40 ꢀC for 2 h, then quenched
with TEA (20 mL) and processed as described for 28. The
residue was subjected to a column of Bio-Beads SX-4 (tolu-
ene/EtOAc, 1:1), then PTLC (toluene/EtOAc, 5:1) to afford
18 (28 mg, 45%) as a white solid: Rf 0.48 (toluene/EtOAc,
1
5:1); H NMR (400 MHz, CDCl3) d 8.00–6.63 (m, 42H),
6.14 (t, 1H, J¼10.0 Hz), 5.86 (t, 1H, J¼10.0 Hz), 5.82

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I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
(d, 1H, J¼3.6 Hz), 5.45 (t, 1H, J¼10.0 Hz), 5.40 (s, 1H),
5.27 (t, 1H, J¼10.0 Hz), 5.19–5.02 (m, 2H), 4.93 (d, 1H,
J¼3.2 Hz), 4.65–4.16 (m, 9H), 3.94 (dd, 1H, J¼3.7,
10.5 Hz), 3.83, 3.81, 3.65 (dd, 1H, J¼3.2, 10.0 Hz), 3.58
(t, 1H, J¼9.3 Hz), 3.50 (s, 3H), 2.28 (s, 3H); 13C NMR
(100 MHz, CDCl3) d 165.94, 165.66, 165.20, 165.10,
164.85, 163.40, 163.13, 138.15, 137.38, 136.87, 133.01,
132.86, 131.83, 131.68, 129.64, 129.59, 129.52, 129.15,
128.58, 128.43, 128.26, 128.15, 128.11, 127.74, 127.45,
126.03, 122.36, 121.64, 121.33, 113.53, 113.37, 95.95,
95.03, 86.05, 81.41, 78.74, 76.60, 75.54, 74.57, 70.98,
70.83, 69.98, 69.52, 69.17, 68.47, 67.84, 67.77, 62.93,
62.21, 55.40, 55.07, 13.01; MALDI-TOF mass m/z calcd
for C85H80O24SNa 1539.5 (M+Na)+, found 1540.2.
Teflon reaction vessel in DMF (2 mL) containing 10% HF/
pyridine. It was compressed to 1.0 GPa and left at 30 ꢀC
for 24 h and the resultant mixture was diluted with EtOAc
and washed successively with aq NaHCO3 and brine, succes-
sively. The organic layer was dried over MgSO4 and evapo-
rated in vacuo. The residue was subjected to a silica gel
column chromatography (hexane/EtOAc 10:1 to 1:2) to
1
give 54 (1.99 g, 95%) as a colorless amorphous: H NMR
(400 MHz, CDCl3) d 7.79–6.71 (m, 83H), 5.59 (m, 1H),
5.41–5.11 (m, 16H), 5.01–4.91 (m, 5H), 4.86–4.77 (m,
7H), 4.69–4.14 (m, 23H), 4.33–4.23 (m, 57H), 3.48 (m,
1H), 3.26 (m, 1H), 2.07–3.14 (m, 2H), 2.11 (s, 3H), 2.09
(s, 3H), 2.07 (s, 3H), 2.06 (s, 3H), 2.04 (s, 6H), 2.02 (s,
3H), 2.00 (s, 3H), 2.00 (s, 3H), 1.99 (s, 3H), 1.98 (s, 3H),
1.97 (s, 3H), 1.96 (s, 3H), 1.96 (s, 3H), 1.91 (s, 3H), 1.90
(s, 3H); 13C NMR (100 MHz, CDCl3) d 170.71, 170.45,
170.14, 169.96, 169.91, 169.58, 169.48, 169.38, 169.25,
169.17, 168.94, 167.90, 167.29, 138.63, 138.43, 138.29,
138.23, 138.10, 138.06, 137.90, 137.76, 137.66, 133.61,
133.35, 131.60, 131.26, 128.42, 128.23, 128.11, 128.06,
127.88, 127.77, 127.70, 127.65, 127.59, 127.50, 127.36,
127.27, 127.07, 126.72, 123.29, 123.00, 117.00, 101.14,
100.77, 99.29, 99.18, 98.68, 98.56, 98.33, 96.96, 96.86,
79.90, 79.71, 78.91, 78.00, 77.53, 77.20, 76.23, 75.74,
75.05, 74.81, 74.70, 74.60, 74.43, 74.29, 74.03, 73.34,
73.21, 73.05, 72.86, 72.76, 72.67, 72.59, 72.50, 72.02,
71.87, 71.56, 71.47, 70.83, 70.07, 69.87, 69.63, 69.57,
69.39, 69.18, 69.05, 68.90, 68.71, 68.54, 68.67, 67.99,
67.21, 66.28, 66.11, 65.88, 65.48, 62.00, 61.89, 61.43,
56.56, 55.61, 20.95, 20.85, 20.71, 20.68, 20.64, 20.60,
20.54; MALDI-TOF mass m/z calcd for C222H244N2NaO74
4144.5 (M+Na)+, found 4144.2; Anal. Calcd for
C222H244N2O74: C, 64.65; H, 5.96; N, 0.68. Found: C,
64.44; H, 5.97; N, 0.65.
4.1.24. Compound 53. A mixture of AgOTf (465 mg,
1.81 mmol), Cp2HfCl2 (344 mg, 0.906 mmol), and molecu-
˚
lar sieves 4 A (6 g) in dry toluene (50 mL) was stirred at
room temperature for 30 min, then cooled at ꢁ30 ꢀC. A
solution of donor 12 (1.45 g, 0.906 mmol) and acceptor 11
(1.66 g, 0.604 mmol) in dry toluene (10 mL) was added
dropwise over 5 min. The mixture was stirred at ꢁ10 ꢀC
for 4 h. The reaction was quenched with TEA (1 mL) and
processed as described for 28. The residue was subjected
to a silica gel column chromatography (hexane/EtOAc, 1:1
to 2:3) to afford 53 (2.27 g, 87%) as a colorless amorphous:
1H NMR (400 MHz, CDCl3) d 7.75–6.70 (m, 93H), 5.56 (m,
1H), 5.39–5.20 (m, 15H), 4.98–3.20 (m, 96H), 3.10 (m, 1H),
2.11 (s, 3H), 2.09 (s, 3H), 2.06 (s, 3H), 2.02 (s, 6H), 2.01 (s,
3H), 1.98 (s, 12H), 1.97 (s, 3H), 1.96 (s, 3H), 1.94 (s, 3H),
1.91 (s, 3H), 1.89 (s, 3H), 1.05 (s, 9H); 13C NMR
(100 MHz, CDCl3) d 170.71, 170.45, 170.14, 169.96,
169.91, 169.58, 169.48, 169.38, 169.25, 169.17, 168.94,
167.90, 167.29, 138.63, 138.43, 138.29, 138.23, 138.18,
138.10, 138.06, 137.90, 137.76, 137.66, 133.61, 133.35,
131.60, 131.26, 128.42, 128.23, 128.11, 128.06, 127.86,
127.77, 127.70, 127.65, 127.59, 127.56, 127.50, 127.36,
127.27, 127.07, 126.72, 123.29, 123.00, 117.00, 101.11,
99.90, 99.31, 99.08, 98.62, 96.91, 79.79, 79.45, 79.17,
78.48, 78.25, 75.88, 75.26, 75.03, 74.86, 74.58, 74.42,
74.34, 74.21, 73.37, 73.19, 73.07, 72.76, 72.58, 72.47,
72.25, 72.18, 71.60, 71.15, 70.04, 69.83, 69.58, 69.51,
69.31, 69.11, 69.00, 68.80, 68.72, 68.55, 68.26, 68.05,
67.93, 67.43, 67.27, 67.09, 66.22, 66.10, 65.87, 65.45,
62.02, 61.83, 61.53, 56.49, 55.56, 38.68, 30.33, 28.88,
27.15, 23.73, 22.94, 21.42, 20.87, 20.81, 20.76, 20.63,
20.60, 20.57, 19.34, 14.03, 10.95; MALDI-TOF mass
m/z calcd for C236H260O73N2Na 4343.6 (M+Na)+, found
4343.5; Anal. Calcd for C236H260N2O73Si: C, 65.60; H,
6.07; N, 0.65. Found: C, 65.56; H, 6.12; N, 0.56.
4.1.26. Compound 55. A mixture of AgOTf (80 mg,
0.31 mmol), Cp2HfCl2 (52 mg, 0.14 mmol), and molecular
˚
sieves 4 A (2 g) in dry toluene (4 mL) was stirred at room
temperature for 30 min, then cooled at ꢁ30 ꢀC. To a solution
of 13 (128 mg, 0.125 mmol) and 11 (208 mg, 0.0759 mmol)
in dry toluene (15 mL) the mixture was added dropwise over
5 min. The mixture was stirred at ꢁ10 ꢀC for 12 h. The mix-
ture was processed as described for 53 and purified by PTLC
(toluene/EtOAc, 3:1) to afford 55 (219 mg, 77%) as a color-
1
less amorphous: H NMR (400 MHz, CDCl3) d 7.76–6.68
(m, 93H), 5.56 (m, 1H), 5.35–5.14 (m, 10H), 4.99–3.23
(m, 98H), 3.13 (m, 1H), 2.12 (s, 3H), 2.08 (s, 3H), 2.03 (s,
3H), 2.02 (s, 3H), 2.01 (s, 3H), 1.99 (s, 3H), 1.97 (s, 3H),
1.96 (s, 3H), 1.95 (s, 3H), 1.94 (s, 3H), 1.92 (s, 3H), 1.08
(s, 9H); 13C NMR (100 MHz, CDCl3) d 170.23, 169.41,
169.34, 138.53, 138.44, 138.41, 138.19, 138.16, 138.05,
138.00, 137.97, 137.92, 137.86, 137.82, 137.77, 137.60,
135.95, 135.87, 133.48, 133.20, 129.33, 128.28, 128.19,
128.09, 128.06, 128.00, 127.95, 127.86, 127.82, 127.70,
127.64, 127.50, 127.32, 127.30, 127.22, 127.19, 127.16,
127.07, 126.83, 126.73, 126.58, 116.84, 100.91, 99.80,
99.49, 99.26, 97.51, 96.87, 79.74, 79.38, 78.20, 75.28,
75.03, 74.89, 74.40, 74.32, 73.91, 73.42, 73.10, 72.80,
72.55, 72.36, 72.17, 71.43, 71.08, 70.98, 69.35, 69.32,
69.21, 69.01, 68.52, 68.32, 67.98, 67.38, 67.06, 66.20,
65.93, 65.80, 62.20, 62.14, 56.49, 55.58, 20.92, 20.85,
20.76, 20.71, 20.68, 19.39; MALDI-TOF mass m/z calcd
for C212H228O74N2SiNa 3764.5 (M+Na)+, found 3764.4;
4.1.25. Compound 54. To a stirred solution of compound 53
(2.27 g, 0.525 mmol) and DMAP (6.4 mg) in pyridine
(40 mL) was added acetic anhydride (20 mL). The mixture
was stirred at 50 ꢀC for 12 h. To the mixture was added
methanol (20 mL) and volatiles were removed by evapora-
tion in vacuo. The residue was diluted with EtOAc and
washed successively with aq CuSO4, brine, aq NaHCO3,
and brine. The organic layer was dried (MgSO4) and evapo-
rated in vacuo. The residue was subjected to a silica gel col-
umn chromatography (hexane/EtOAc 5:1 to 1:3) to give the
acetylated undecasaccharide (2.22 g, 97%). The acetylated
compound (2.22 g, 0.509 mmol) was dissolved in a 3 mL

I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
8273
Anal. Calcd for C212H228N2O57Si: C, 68.01; H, 6.14; N,
0.75. Found: C, 67.77; H, 6.19; N, 0.69.
1.89 (s, 6H), 1.87 (s, 3H); 13C NMR (100 MHz, CDCl3)
d 170.36–169.30, 138.35–127.00, 116.95, 101.15, 100.59,
99.30, 98.69, 98.29, 97.49, 96.91, 96.80, 79.89, 79.65,
78.97, 77.95, 77.64, 77.14, 77.02, 76.67, 76.18, 75.71,
75.05, 74.86, 74.79, 74.62, 74.40, 74.26, 74.01, 73.05,
72.86, 72.74, 72.65, 72.56, 71.86, 71.52, 71.45, 71.14,
70.85, 70.03, 69.58, 69.15, 69.02, 68.96, 68.76, 68.65,
68.49, 68.39, 67.97, 67.86, 67.28, 66.18, 66.10, 65.87,
65.51, 62.21, 62.05, 61.43, 56.58, 55.62, 21.54, 20.90,
20.84, 20.76, 20.68; MALDI-TOF mass m/z calcd for
C210H228O66N2Na 3856.5 (M+Na)+, found 3857.6; Anal.
Calcd for C210H228N2O66: C, 65.75; H, 5.99; N, 0.73. Found:
C, 65.76; H, 6.19; N, 0.68.
4.1.27.Compound56.Compound 55 (205 mg, 0.0548 mmol)
was acetylated and desilylated as described for 54. The
residue was subjected to a PTLC (toluene/EtOAc 2:1) to
give 56 (168 mg, 86%) as a colorless amorphous: 1H NMR
(400 MHz, CDCl3) d 7.78–6.72 (m, 83H), 5.58 (m, 1H),
5.38 (d, 1H, J¼2.9 Hz), 5.34–3.49 (m, 93H), 3.46–3.31
(m, 2H), 3.34–3.18 (m, 3H), 2.05 (s, 3H), 2.04 (s, 3H),
1.99 (s, 6H), 1.97 (s, 3H), 1.94 (s, 3H), 1.93 (s, 3H), 1.92
(s, 3H), 1.91 (s, 3H), 1.87 (s, 3H); 13C NMR (100 MHz,
CDCl3) d 170.31, 170.03, 169.48, 169.44, 163.31, 169.28,
169.23, 167.77, 167.15, 138.54, 138.35, 138.32, 138.21,
138.12, 138.05, 137.98, 137.94, 137.84, 137.81, 137.71,
137.65, 133.61, 133.50, 131.31, 131.16, 128.34, 128.24,
128.14, 128.11, 128.05, 128.03, 127.98, 127.87, 127.80,
127.77, 127.69, 127.65, 127.61, 127.56, 127.52, 127.46,
127.41, 127.31, 127.27, 127.22, 127.19, 127.15, 127.01,
126.78, 126.65, 123.32, 122.89, 116.95, 101.08, 100.68,
99.18, 98.32, 97.45, 96.92, 96.77, 79.85, 79.53, 78.78,
77.95, 77.82, 77.20, 76.20, 75.69, 75.04, 74.85, 74.78,
74.69, 74.55, 74.38, 74.29, 74.01, 73.86, 73.33, 73.17,
73.04, 72.85, 72.72, 72.64, 72.49, 72.00, 71.83, 71.63,
71.50, 71.43, 71.18, 70.85, 69.39, 69.26, 69.17, 69.03,
68.95, 68.69, 68.60, 68.37, 67.98, 67.86, 67.22, 66.71,
66.23, 65.93, 65.68, 62.24, 62.00, 56.54, 55.61, 21.04,
20.94, 20.90, 20.83, 20.75, 20.66; MALDI-TOF mass
m/z calcd for C198H212O58N2Na 3568.4 (M+Na)+, found
3568.1; Anal. Calcd for C198H212N2O58: C, 67.03; H, 6.02;
N, 0.79. Found: C, 66.86; H, 6.00; N, 0.75.
4.1.30. Compound 59. A mixture of AgOTf (103 mg,
0.401 mmol), Cp2HfCl2 (77.7 mg, 0.201 mmol), and molec-
˚
ular sieves 4 A (4 g) in dry toluene (5 mL) was stirred at
room temperature for 30 min, then cooled at ꢁ30 ꢀC. To a
solution of 15 (381 mg, 0.291 mmol) and 11 (550 mg,
0.201 mmol) in dry toluene (45 mL) the mixture was added
dropwise over 5 min. The reaction mixture was stirred
at ꢁ10 ꢀC for 3 h and processed as described for 53. The
residue was subjected to a silica gel column chromatography
(hexane/EtOAc, 1:1 to 2:3) to afford 59 (682 mg, 84%) as
a colorless amorphous: 1H NMR (400 MHz, CDCl3)
d 7.68–6.70 (m, 93H), 5.57 (m, 1H), 5.37–5.12 (m, 12H),
5.01–3.43 (m, 89H), 3.30 (m, 2H), 3.22 (m, 2H), 3.11 (m,
1H), 2.00 (s, 6H), 1.92 (s, 9H), 1.92 (s, 3H), 1.90 (s, 6H),
1.88 (s, 3H), 1.87 (s, 3H), 1.84 (s, 3H), 1.83 (s, 3H), 1.00
(s, 9H); 13C NMR (100 MHz, CDCl3) d 170.60, 170.13,
170.01, 169.35, 169.30, 169.29, 169.07, 138.49, 138.45,
138.42, 138.23, 138.21, 138.09, 138.03, 137.84, 137.79,
136.00, 135.92, 133.54, 128.31, 128.22, 128.13, 128.05,
128.00, 127.90, 127.83, 127.75, 127.73, 127.61, 127.56,
127.37, 127.35, 127.31, 127.21, 127.13, 127.10, 126.91,
126.80, 126.60, 126.45, 116.91, 101.01, 99.97, 99.41,
99.09, 98.41, 96.93, 96.87, 79.78, 79.50, 79.10, 78.28,
77.71, 77.56, 77.20, 75.86, 75.32, 75.05, 74.89, 74.48,
74.34, 73.89, 73.41, 73.22, 73.16, 72.81, 72.61, 72.47,
72.21, 71.77, 71.53, 71.29, 71.14, 69.94, 69.77, 69.64,
69.38, 69.21, 69.05, 68.91, 68.63, 68.56, 68.37, 68.02,
67.48, 67.32, 67.01, 66.29, 66.15, 65.94, 65.81, 62.11,
61.90, 56.53, 55.63, 27.26, 20.99, 20.91, 20.84, 20.73,
19.45; MALDI-TOF mass m/z calcd for C224H244O65N2SiNa
4053.5 (M+Na)+, found 4053.8; Anal. Calcd for
C224H244N2O65Si: C, 66.72; H, 6.10; N, 0.69. Found: C,
66.51; H, 6.13; N, 0.66.
4.1.28. Compound 57. A mixture of AgOTf (84.9 mg,
0.330 mmol), Cp2HfCl2 (62.8 mg, 0.165 mmol), and molec-
˚
ular sieves 4 A (5 g) in dry toluene (8 mL) was stirred at
room temperature for 30 min, then cooled at ꢁ30 ꢀC. To a
solution of 14 (146 mg, 0.111 mmol) and 11 (268 mg,
0.0978 mmol) in dry toluene (15 mL) the mixture was added
dropwise over 5 min and stirred at ꢁ10 ꢀC for 3 h. The reac-
tion mixture was processed as described for 53 and purified
by silica gel column chromatography (toluene/EtOAc, 5:1 to
1:1) to afford 57 (295 mg, 75%) as a colorless amorphous:
1H NMR (400 MHz, CDCl3) d 7.75–6.72 (m, 93H), 5.64
(m, 1H), 5.34–3.22 (m, 104H), 3.14 (m, 1H), 2.16 (s, 3H),
2.09 (s, 3H), 2.08 (s, 3H), 2.05 (s, 3H), 2.04 (s, 6H), 2.03
(s, 6H), 2.02 (s, 3H), 1.97 (s, 3H), 1.96 (s, 3H), 1.94 (s,
3H), 1.12 (s, 9H); 13C NMR (100 MHz, CDCl3) d 170.36–
169.30, 138.45–126.76, 116.91, 101.08, 99.68, 99.58,
99.25, 98.62, 97.79, 97.51, 96.94, 96.85, 56.53, 55.58,
27.22, 20.93, 20.84, 20.76, 20.70, 20.66, 19.42, 14.29;
MALDI-TOF mass m/z calcd for C224H244O65N2SiNa
4052.6 (M+Na)+, found 4053.7; Anal. Calcd for
C224H244N2O65Si: C, 66.72; H, 6.10; N, 0.69. Found: C,
66.50; H, 6.13; N, 0.61.
4.1.31. Compound 60. Compound 59 (303 mg,
0.0751 mmol) was acetylated and desilylated as described
for 54. Purification by silica gel column chromatography
(toluene/EtOAc, 5:1 to 1:1) afforded 60 (285 mg, 97%) as
a colorless amorphous: 1H NMR (400 MHz, CDCl3)
d 7.78–6.73 (m, 83H), 5.58 (m, 1H), 5.38–4.40 (m, 46H),
4.32–3.45 (m, 48H), 3.67 (m, 1H), 3.31 (m, 1H), 3.23 (m,
1H), 3.17 (m, 1H), 2.11 (s, 3H), 2.08 (s, 3H), 2.05 (s, 3H),
2.04 (s, 3H), 2.00 (s, 3H), 1.99 (s, 3H), 1.98 (s, 3H), 1.98
(s, 3H), 1.97 (s, 3H), 1.95 (s, 3H), 1.94 (s, 3H), 1.91 (s,
3H); 13C NMR (100 MHz, CDCl3) d 170.64, 170.08,
169.73, 169.50, 169.43, 169.34, 169.15, 138.53, 138.36,
138.31, 138.18, 138.13, 138.05, 138.02, 137.98, 137.82,
137.76, 137.66, 133.54, 128.37, 128.28, 128.19, 128.14,
128.09, 128.07, 128.02, 127.95, 127.87, 127.82, 127.13,
4.1.29. Compound 58. Compound 57 (275 mg,
0.0682 mmol) was acetylated and desilylated as described
for 54. Purification by silica gel column chromatography
(hexane/EtOAc, 5:1 to 1:1) gave 58 (182 mg, 70%) as a
1
colorless amorphous: H NMR (400 MHz, CDCl3) d 7.78–
6.73 (m, 83H), 5.58 (m, 1H), 5.54 (d, 1H, J¼4.8 Hz),
5.40–3.12 (m, 107H), 2.10 (s, 3H), 2.03 (s, 9H), 2.02 (s,
3H), 1.99 (s, 6H), 1.98 (s, 3H), 1.97 (s, 3H), 1.96 (s, 3H),

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I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
127.66, 127.60, 127.56, 127.42, 127.32, 127.30, 127.23,
127.12, 127.03, 126.68, 126.42, 117.00, 101.07, 100.78,
99.17, 98.33, 97.11, 96.94, 96.82, 79.90, 79.62, 78.76,
77.98, 77.74, 77.55, 77.20, 76.23, 75.73, 75.08, 74.86,
74.64, 74.42, 74.34, 74.06, 73.68, 73.38, 73.21, 73.07,
72.86, 72.79, 72.66, 72.55, 72.47, 72.03, 71.88, 71.68,
71.54, 71.47, 71.27, 70.85, 69.96, 69.63, 69.44, 69.22,
69.05, 68.91, 68.73, 68.65, 68.55, 68.37, 68.21, 68.00,
67.18, 66.28, 66.17, 65.90, 65.77, 62.10, 62.00, 61.94,
56.53, 55.64, 21.06, 20.96, 20.87, 20.80, 20.76, 20.73;
MALDI-TOF mass m/z calcd for C210H228O66N2Na 3856.5
(M+Na)+, found 3856.6; Anal. Calcd for C210H228N2O66:
C, 65.75; H, 5.99; N, 0.73. Found: C, 65.76; H, 5.89; N, 0.75.
63 (555 mg, 93%) as a colorless amorphous: 1H NMR
(400 MHz, CDCl3) d 7.79–6.64 (m, 92H), 5.71 (t, 1H,
J¼9.8 Hz), 5.58 (m, 1H), 5.41 (s, 1H), 5.38 (br d, 1H,
J¼2.8 Hz), 5.36–3.42 (m, 117H), 3.33 (m, 1H), 3.23 (m,
1H), 3.17 (m, 2H), 2.10 (s, 3H), 2.07 (s, 3H), 2.06 (s, 3H),
2.04 (s, 3H), 2.03 (s, 3H), 2.02 (s, 3H), 2.00 (s, 3H), 1.99
(s, 3H), 1.98 (s, 9H), 1.97 (s, 3H), 1.96 (s, 3H), 1.95 (s,
3H), 1.90 (s, 3H), 1.88 (s, 3H), 1.71 (s, 9H); MALDI-TOF
mass m/z calcd for C248H274N2O81Na 4601.8 (M+Na)+,
found 4601.7; Anal. Calcd for C248H274N2O81: C, 65.05;
H, 6.03; N, 0.61. Found: C, 64.51; H, 5.97; N, 0.58.
4.1.35. Compound 64. A mixture of 56 (36.7 mg,
0.0103 mmol), 16 (36.6 mg, 0.0728 mmol), DTBMP
˚
(30 mg, 0.14 mmol), and molecular sieves 4 A (1.0 g) in
4.1.32. Compound 61. A mixture of 18 (28.1 mg,
0.0185 mmol), 54 (49.8 mg, 0.0120 mmol), DTBMP (38 mg,
˚
0.012 mmol), and molecular sieves 4 A (500 mg) in dry
dry ClCH2CH2Cl (2 mL) and cyclohexane (4 mL) was
stirred at room temperature for 30 min, then added 1 M
MeOTf (0.100 mL, 0.100 mmol) in ClCH2CH2Cl. The reac-
tion mixture was stirred at 45 ꢀC for 19 h and processed as
described for 61. The mixture was subjected to a PTLC
(toluene/EtOAc, 3:4) to afford 64 (35.6 mg, 86%) as a color-
ClCH2CH2Cl (1 mL) and cyclohexane (4 mL) was stirred
at room temperature for 2 h, then added 1 M MeOTf
(0.15 mL, 0.15 mmol) in ClCH2CH2Cl. The reaction mix-
ture was stirred at 50 ꢀC for 12 h. The reaction was quenched
with TEA (0.1 mL). The mixture was diluted with EtOAc
and filtered through Celite. The filtrate was washed with
aq NaHCO3, and brine. The organic layer was dried over
Na2SO4 and concentrated in vacuo. The residue was sub-
jected to a column of Bio-Beads SX-4 (toluene/EtOAc,
1:1), then PTLC (toluene/EtOAc, 1:2) to afford 61
(38.1 mg, 57%) as a colorless amorphous: Rf 0.75 (tolu-
1
less amorphous: H NMR (400 MHz, CDCl3) d 7.75–6.42
(m, 92H), 5.72 (t, 1H, J¼9.6 Hz), 5.58 (m, 1H), 5.42 (s,
1H), 5.38 (d, 1H, J¼3.2 Hz), 5.34–3.45 (m, 102H), 3.33
(m, 2H), 3.23 (m, 1H), 3.18 (m, 2H), 2.05 (s, 3H), 2.03 (s,
3H), 1.99 (s, 3H), 1.98 (s, 3H), 1.97 (s, 3H), 1.95 (s, 3H),
1.93 (s, 3H), 1.92 (s, 3H), 1.91 (s, 3H), 1.88 (s, 3H), 1.17
(s, 9H); MALDI-TOF mass m/z calcd for C224H242N2O65Na
(M+Na)+ 4022.6, found 4022.5; Anal. Calcd for
C224H242N2O65: C, 67.22; H, 6.09; N, 0.70. Found: C,
67.43; H, 6.35; N, 0.56.
1
ene/EtOAc, 2:3); H NMR (400 MHz, CDCl3) d 7.97–6.56
(m, 125H), 6.10 (t, 1H, J¼10.0 Hz), 5.76 (t, 1H,
J¼10.0 Hz), 5.66 (d, 1H, J¼3.2 Hz), 5.55–5.47 (m, 1H),
5.43 (t, 1H, J¼10.0 Hz), 2.03, 2.00, 1.99, 1.96, 1.95, 1.93,
1.92, 1.91, 1.91, 1.90, 1.89, 1.88; MALDI-TOF mass m/z
calcd for C306H320N2O98Na 5612.9 (M+Na)+, found 5611.3.
4.1.36. Compound 65. A mixture of 58 (71.2 mg,
0.0186 mmol), 16 (37.9 mg, 0.0754 mmol), DTBMP
˚
(70.3 mg, 0.570 mmol), and molecular sieves 4 A (2.0 g)
4.1.33. Compound 62. A mixture of 17 (30.6 mg,
0.0321 mmol), 54 (77.4 mg, 0.0188 mmol), DTBMP
in dry ClCH2CH2Cl (4 mL) and cyclohexane (10 mL) was
stirred at room temperature for 30 min, then added 1 M
MeOTf (0.400 mL, 0.400 mmol) in ClCH2CH2Cl. The reac-
tion mixture was stirred at 45 ꢀC for 12 h. The mixture was
processed as described for 61 and purified by PTLC
(toluene/EtOAc, 3:4) to afford 65 (72.5 mg, 91%) as a color-
˚
(52 mg, 0.26 mmol), and molecular sieves 4 A (500 mg) in
dry ClCH2CH2Cl (3 mL) and cyclohexane (9 mL) was
stirred at room temperature for 30 min then added 1 M
MeOTf (0.2 mL, 0.2 mmol) in ClCH2CH2Cl. The reaction
mixture was stirred at 50 ꢀC for 12 h and processed as
described for 61. Purification by PTLC (toluene/EtOAc,
5:4) afforded 62 (80.4 mg, 85%) as a colorless amorphous:
1H NMR (400 MHz, CDCl3) d 8.00–6.61 (m, 113H), 5.94
(t, 1H, J¼9.0 Hz), 5.65 (d, 1H, J¼4.0 Hz), 5.63–5.54 (m,
1H), 5.42–3.45 (m, 124H), 3.30 (m, 1H), 3.25–3.18 (m,
2H), 2.10 (s, 3H), 2.07 (s, 3H), 2.05 (s, 3H), 2.04 (s, 3H),
2.03 (s, 3H), 2.02 (s, 3H), 2.00 (s, 3H), 1.99 (s, 3H), 1.98
(s, 3H), 1.98 (s, 6H), 1.97 (s, 3H), 1.96 (s, 3H), 1.95 (s,
3H), 1.90 (s, 3H), 1.89 (s, 3H); MALDI-TOF mass m/z calcd
for C276H292N2O87Na 5052.2 (M+Na)+, found 5052.4; Anal.
Calcd for C276H292N2O87: C, 65.91; H, 5.85; N, 0.56. Found:
C, 65.51; H, 5.80; N, 0.49.
1
less amorphous: H NMR (400 MHz, CDCl3) d 7.57–6.63
(m, 92H), 5.71 (t, 1H, J¼10.0 Hz), 5.90 (m, 1H), 5.41 (br
s, 2H), 5.33–3.14 (m, 114H), 2.10 (s, 3H), 2.03 (s, 9H),
2.02 (s, 3H), 2.00 (s, 3H), 1.99 (s, 6H), 1.98 (s, 3H), 1.96
(s, 3H), 1.90 (s, 3H), 1.88 (s, 3H), 1.16 (s, 9H); MALDI-
TOF mass m/z calcd for C236H258N2O73Na 4310.6
(M+Na)+, found 4311.7; Anal. Calcd for C236H258N2O73:
C, 66.06; H, 6.06; N, 0.65. Found: C, 65.81; H, 5.92; N, 0.59.
4.1.37. Compound 66. A mixture of 60 (276 mg,
0.0719 mmol), 16 (111 mg, 0.221 mmol), DTBMP (83.5 mg,
˚
0.407 mmol), and molecular sieves 4 A (3.0 g) in dry
ClCH2CH2Cl (10 mL) and cyclohexane (20 mL) was stirred
at room temperature for 30 min, then added 1 M MeOTf
(0.288 mL, 0.288 mmol) in ClCH2CH2Cl. The reaction mix-
ture was stirred at 45 ꢀC for 60 h and processed as described
for 61. Purification by PTLC (toluene/EtOAc, 1:2) afforded
66 (245 mg, 79%) as a colorless amorphous: 1H NMR
(400 MHz, CDCl3) d 7.78–6.64 (m, 92H), 5.72 (t, 1H,
J¼9.3 Hz), 5.58 (m, 1H), 5.42 (s, 1H), 5.38–3.30 (m,
112H), 3.20 (m, 3H), 2.08 (s, 3H), 2.05 (s, 3H), 2.04 (s,
3H), 2.00 (s, 3H), 1.99 (s, 3H), 1.98 (s, 3H), 1.97 (s, 3H),
4.1.34. Compound 63. A mixture of 54 (500 mg,
0.121 mmol), 16 (244 mg, 0.485 mmol), DTBMP (117 mg,
˚
0.558 mmol), and molecular sieves 4 A (5 g) in dry
ClCH2CH2Cl (11 mL) and cyclohexane (55 mL) was stirred
at room temperature for 30 min, then added 1 M MeOTf
(0.68 mL, 0.68 mmol) in ClCH2CH2Cl. The reaction mix-
ture was stirred at 50 ꢀC for 12 h and processed as described
for 61. Purification by PTLC (toluene/EtOAc, 1:2) afforded

I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
8275
1.96 (s, 3H), 1.95 (s, 3H), 1.95 (s, 3H), 1.93 (s, 3H), 1.91 (s,
3H), 1.71 (s, 9H); MALDI-TOF mass m/z calcd for
C236H258N2O73Na 4310.6 (M+Na)+, found 4311.2; Anal.
Calcd for C236H258N2O73: C, 66.06; H, 6.06; N, 0.65. Found:
C, 65.94; H, 6.02; N, 0.59.
(br s, 1H), 4.73 (br s, 1H), 4.45 (d, 1H, J¼7.6 Hz), 4.36 (d,
1H, J¼7.6 Hz), 4.10 (br s, 1H), 4.02–3.38 (m, 54H), 1.94
(s, 3H), 1.89 (s, 3H), 1.41 (m, 2H), 0.73 (t, 3H, J¼7.3 Hz);
MALDI-TOF mass m/z calcd for C67H114N2O51Na 1785.6
(M+Na)+, found 1785.2.
4.1.38. Compound 1 (M9). A solution of undecasaccharide
51 (28.7 mg, 0.00696 mmol) in n-butanol (2 mL) containing
0.5 mL ethylenediamine was stirred at 90 ꢀC for 15 h. Vola-
tiles were removed by evaporation in vacuo and the residue
was dissolved in pyridine (0.3 mL). The solution was treated
with Ac2O (0.2 mL) at 0 ꢀC for 24 h and evaporated
in vacuo. The residue was dissolved in MeOH (5 mL) and
1 N NaOMe/MeOH (0.1 mL) was added at 0 ꢀC. The mix-
ture was stirred at 60 ꢀC for 12 h, neutralized with Amber-
lyst 15 (H+) resin, and evaporated in vacuo. The residue
was applied to a gel filtration chromatography (Sephadex
LH20, CHCl3/MeOH, 1:1) to collect the deacetylated com-
pound and evaporated in vacuo. The residue was hydro-
genated over Pd(OH)2/C (20 wt %, 5 mg) in 60% aq
AcOH (5 mL) at room temperature for 24 h. The mixturewas
filtered through Celite. The filtrate was concentrated
in vacuo. The residue was subjected to a gel filtration
(Sephadex LH 20, H2O) to afford 1 (7.3 mg, 54%) as a white
powder: 1H NMR (400 MHz, D2O) d 5.27 (br s, 1H), 5.20 (br
s, 1H), 5.17 (br s, 1H), 5.01 (br s, 1H), 4.92 (br s, 1H), 4.90
(br s, 2H), 4.73 (br s, 1H), 4.45 (d, 1H, J¼7.6 Hz), 4.36 (d,
1H, J¼7.8 Hz), 4.09 (br s, 1H), 4.02–3.37 (m, 67H), 1.93
(s, 3H), 1.88 (s, 3H), 1.40 (m, 2H), 0.72 (t, 3H, J¼7.3 Hz);
MALDI-TOF mass m/z calcd for C73H124N2O56Na 1947.7
(M+Na)+, found 1947.7.
4.1.41. Compound 4 (M8B). Decasaccharide 60 (28.7 mg,
0.00748 mmol) was subjected to a series of reactions in
a manner as described for 1. The mixture was subjected to
a Sep-Pak C18 cartridge (500 mg, Waters) (H2O only to
H2O/MeOH, 20:1) to give 4 (6.2 mg, 47%) as a white
1
powder. H NMR (400 MHz, D2O) d 5.20 (br s, 1H), 5.17
(br s, 1H), 5.01 (br s, 1H), 4.95 (br s, 1H), 4.90 (br s, 2H),
4.73 (br s, 1H), 4.45 (br d, 1H, J¼7.5 Hz), 4.36 (d, 1H,
J¼8.0 Hz), 4.09 (br d, 1H, J¼2.4 Hz), 4.01–3.36 (m,
54H), 1.93 (s, 3H), 1.89 (s, 3H), 1.41 (m, 2H), 0.72 (t, 3H, J¼
7.3 Hz); MALDI-TOF mass m/z calcd for C67H114N2O51Na
1785.6 (M+Na)+, found 1786.3.
4.1.42. Compound 5 (G3M9). Tetradecasaccharide 61
(26.7 mg, 0.00477 mmol) was subjected to a series of reac-
tions in a manner as described for 1. The mixture was puri-
fied by Sep-Pak C18 cartridge (500 mg, Waters, H2O only to
H2O/MeOH, 20:1), then with HPLC (column Fluofix, H2O)
to give 5 (6.5 mg, 56%) as a white powder: 1H NMR
(400 MHz, D2O) d 5.39 (d, 1H, J¼2.4 Hz), 5.27 (br s, 1H),
5.20 (br s, 1H), 5.17 (br s, 1H), 5.13 (br d, 1H, J¼2.4 Hz),
5.04 (br d, 1H, J¼3.2 Hz), 5.01 (br s, 1H), 4.12 (br s, 1H),
4.90 (br s, 2H), 4.76 (br s, 1H), 4.45 (d, 1H, J¼7.2 Hz),
4.36 (d, 1H, J¼6.8 Hz), 4.10 (br s, 2H), 4.02–3.30
(m, 84H), 1.93 (s, 3H), 1.89 (s, 3H), 1.40 (m, 2H), 0.72
(t, 3H, J¼7.6 Hz); MALDI-TOF mass m/z calcd for
C91H154N2O71Na 2433.8 (M+Na)+, found 2432.8.
4.1.39. Compound 2 (M7). A solution of nonasaccharide 56
(100 mg, 0.0282 mmol) in n-butanol (3 mL) containing
2 mL of ethylenediamine was stirred at 90 ꢀC for 15 h. Vola-
tiles were removed by evaporation in vacuo and the residue
was dissolved in pyridine (3 mL). The solution was treated
with Ac2O (1.5 mL) at 0 ꢀC for 24 h and evaporated in vacuo
to give acetylated compound (quantitative yield). The acetyl-
ated compound (22.6 mg, 0.0067 mmol) was hydrogenated
over Pd(OH)2/C (20 wt %, 20 mg) in a mixture of MeOH
(10 mL) and 60% aq AcOH (5 mL) at room temperature
for 24 h. The mixture was filtered through Celite. The filtrate
was concentrated in vacuo. The residue was dissolved in
MeOH (5 mL) and 1 N NaOMe/MeOH (0.1 mL). The mix-
ture was stirred at 60 ꢀC for 12 h, neutralized with Amber-
lyst 15 (H+) resin, and evaporated in vacuo. The residue
was subjected to a Sep-Pak C18 cartridge (500 mg, Waters)
(H2O only to H2O/MeOH¼20:1) to give 2 (9.1 mg, 84%) as
a white powder: 1H NMR (400 MHz, D2O) d 5.21 (br s, 1H),
5.17 (br s, 1H), 4.96 (br s, 1H), 4.91 (br s, 1H), 4.77 (br s,
1H), 4.73 (br s, 1H), 4.45 (br d, 1H, J¼6.6 Hz), 4.36 (d,
1H, J¼6.8 Hz), 4.10 (br s, 1H), 4.01 (br s, 1H), 3.97–3.37
(m, 54H), 1.93 (s, 3H), 1.89 (s, 3H), 1.41 (m, 2H), 0.73
(t, 3H, J¼7.3 Hz); MALDI-TOF mass m/z calcd for
C61H104N2O46Na 1623.6 (M+Na)+, found 1624.7.
4.1.43. Compound 6 (G2M9). Tridecasaccharide 62
(80.4 mg, 0.0160 mmol) in n-butanol (2 mL) containing
ethylenediamine (1 mL) was stirred at 80 ꢀC for 12 h. Vola-
tiles were removed by evaporation in vacuo and the residue
was dissolved in pyridine (4 mL). The solution was treated
with Ac2O (2 mL) and stirred at 40 ꢀC for 12 h and evapo-
rated in vacuo. The residue was diluted with EtOAc and
washed with 1 N HCl, brine, aq NaHCO3, and brine. The
organic layer was dried (Na2SO4) and evaporated in vacuo.
The residue was dissolved in MeOH (10 mL) and 1 N
NaOMe/MeOH (0.5 mL) was added at 0 ꢀC. The mixture
was stirred at 40 ꢀC for 5 h, neutralized with Amberlyst 15
(H+) resin, and evaporated in vacuo. The residue was
subjected to a PTLC (CHCl3/MeOH, 5:1) to give the de-
acetylated compound (44 mg, 72%). The deacetylated com-
pound (28.8 mg, 0.00745 mmol) was hydrogenated over
Pd(OH)2–C (20 wt %, 20 mg) in 50% aq AcOH (5 mL) at
room temperature for 12 h. The mixture was filtered through
Celite. The filtrate was concentrated in vacuo. The residue
was subjected to a Sep-Pak C18 cartridge (500 mg, Waters)
(H2O only to H2O/MeOH, 20:1) to give 6 (11.0 mg, 65%) as
a white powder: 1H NMR (400 MHz, D2O) d 5.26 (br s, 1H),
5.21 (d, 1H, J¼3.6 Hz), 5.19 (br s, 1H), 5.16 (br s, 1H), 5.12
(d, 1H, J¼3.9 Hz), 5.00 (br s, 1H), 4.90 (br s, 1H), 4.89 (br s,
2H), 4.71 (br s, 1H), 4.57 (br d, 1H, J¼7.0 Hz), 4.35 (br d,
1H, J¼6.8 Hz), 4.09 (br s, 2H), 4.01–3.30 (m, 80H), 1.92
(s, 3H), 1.88 (s, 3H), 1.40 (m, 2H), 0.71 (t, 3H, J¼7.3 Hz);
MALDI-TOF mass m/z calcd for C85H144N2O66Na 2271.8
(M+Na)+, found 2271.4.
4.1.40. Compound 3 (M8C). In a manner as described for 1,
decasaccharide 58 (14.5 mg, 0.00378 mmol) was subjected
to a series of reactions. Purification by gel filtration (Sepha-
dex LH20, H2O) gave compound 3 (6.0 mg, 87%) as a white
powder: 1H NMR (400 MHz, D2O) d 5.27 (br s, 1H), 5.20 (br
s, 1H), 5.17 (br s, 1H), 4.92 (br s, 1H), 4.91 (br s, 1H), 4.77

8276
I. Matsuo et al. / Tetrahedron 62 (2006) 8262–8277
4.1.44. Compound 7 (G1M9). Dodecasaccharide 63
(34.7 mg, 0.00757 mmol) was subjected to a series of reac-
tions in a manner as described for 1. Subsequent purification
by gel filtration (Sephadex LH 20, H2O) afforded compound
Frontier Science Program (RGP0031/2005-C) are acknowl-
edged.
References and notes
1
7 (8.5 mg, 54%) as a white powder: H NMR (400 MHz,
D2O) d 5.27 (br s, 1H), 5.20 (br s, 1H), 5.17 (br s, 1H),
5.12 (d, 1H, J¼3.7 Hz), 5.01 (br s, 1H), 4.92 (br s, 1H),
4.90 (br s, 2H), 4.73 (br s, 1H), 4.46 (d, 1H, J¼7.3 Hz),
4.37 (d, 1H, J¼7.5 Hz), 4.10 (br s, 2H), 4.02–3.24 (m,
71H), 1.94 (s, 3H), 1.89 (s, 3H), 1.41 (m, 2H), 0.73 (t,
3H, J¼7.3 Hz); MALDI-TOF mass m/z calcd for
C79H134N2O61Na 2109.7(M+Na)+, found 2109.4.
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4.1.45. Compound 8 (G1M7). Decasaccharide 64 (35.6 mg,
0.00898 mmol) was subjected to a series of reactions in
a manner as described for 1. The mixture was subjected to
a Sep-Pak C18 cartridge (2 g, Waters) (H2O only to H2O/
MeOH, 10:1) to give 8 (15.1 mg, 84%) as a white powder:
1H NMR (400 MHz, D2O) d 5.21 (br s, 1H), 5.17 (br s,
1H), 5.11 (d, 1H, J¼3.6 Hz), 4.95 (br s, 1H), 4.90 (br s,
1H), 4.77 (br s, 1H), 4.73 (br s, 1H), 4.44 (br d, 1H,
J¼6.6 Hz), 4.36 (d, 1H, J¼7.3 Hz), 4.10 (br s, 2H), 4.01–
3.24 (m, 61H), 1.93 (s, 3H), 1.89 (s, 3H), 1.41 (m, 2H),
0.72 (t, 3H, J¼7.3 Hz); MALDI-TOF mass m/z calcd for
C67H114N2O51Na 1785.6 (M+Na)+, found 1785.6.
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6. Jakob, C. A.; Bodmer, D.; Spirig, U.; Battig, P.; Mardil, A.;
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7. Spiro, R. G. Cell Mol. Life Sci. 2004, 61, 1025–1041.
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11. Ito, Y.; Hagihara, S.; Matsuo, I.; Totani, K. Curr. Opin. Struct.
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14. Kapoor, M.; Srinivas, H.; Kandiah, E.; Gemma, E.; Ellgaard,
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15. Kamiya, Y.; Yamaguchi, Y.; Takahashi, N.; Arata, Y.; Kasai,
K.; Ihara, Y.; Matsuo, Y.; Ito, Y.; Yamamoto, K.; Kato, K.
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4.1.46. Compound 9 (G1M8C). Undecasaccharide 65
(68.8 mg, 0.0160 mmol) was subjected to a series of reac-
tions in a manner as described for 1. The mixture was sub-
jected to a Sep-Pak C18 cartridge (2 g, Waters) (H2O only
to H2O/MeOH, 20:1) to give 9 (20.7 mg, 66%) as a white
1
powder: H NMR (400 MHz, D2O) d 5.27 (br s, 1H), 5.20
(br s, 1H), 5.17 (br s, 1H), 5.11 (d, 1H, J¼4.0 Hz), 4.92
(br s, 1H), 4.90 (br s, 1H), 4.77 (br s, 1H), 4.73 (br s, 1H),
4.45 (d, 1H, J¼7.5 Hz), 4.36 (d, 1H, J¼8.0 Hz), 4.09 (br s,
2H), 3.97–3.21 (m, 67H), 1.94 (s, 3H), 1.89 (s, 3H), 1.41
(m, 2H), 0.73 (t, 3H, J¼7.3 Hz); MALDI-TOF mass m/z
calcd for C73H124N2O56Na 1947.7 (M+Na)+, found 1947.5.
4.1.47. Compound 10 (G1M8B). Undecasaccharide Com-
pound 66 (24.2 mg, 0.00564 mmol) was subjected to a series
of reactions in a manner as described for 1. The mixture was
subjected to a Sep-Pak C18 cartridge (500 mg, Waters) (H2O
only to H2O/MeOH, 20:1) to give 10 (6.3 mg, 58%) as
19. Ito, Y.; Ohnishi, Y.; Ogawa, T.; Nakahara, Y. Synlett 1998,
1102–1104.
1
a white powder: H NMR (400 MHz, D2O) d 5.21 (br s,
1H), 5.17 (br s, 1H), 5.11 (d, 1H, J¼3.9 Hz), 5.01 (br s,
1H), 4.95 (br s, 1H), 4.90 (br s, 2H), 4.68 (br s, 1H), 4.45
(d, 1H, J¼6.8 Hz), 4.36 (d, 1H, J¼7.2 Hz), 4.10 (br s, 2H),
3.01–3.26 (m, 67H), 1.93 (s, 3H), 1.89 (s, 3H), 1.40 (m,
2H), 0.72 (t, 3H, J¼7.3 Hz); MALDI-TOF mass m/z calcd
for C73H124N2O56Na 1947.7 (M+Na)+, found 1948.6.
20. Seifert, J.; Lergenm
2000, 39, 531–534.
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Acknowledgements
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3275.
We thank Ms. Megumi Wada and Mr. Toshinori Kashiwagi
for their contribution at the exploratory stage of this work,
Dr. Teiji Chihara for the operation of the high-pressure
equipment, and Ms. Akemi Takahashi for technical assis-
tance. Financial support from Ministry of Education,
Culture, Sports, Science and Technology [Grant-in-Aid
for Creative Scientific Research (17GS0420) and Encour-
agement of Young Scientists (No. 15780220)] and Human
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