Synthesis of a library of fucopyranosyl-galactopyranosides consisting of a complete set of anomeric configurations and linkage positions

Article abstract of DOI:10.1016/j.carres.2006.03.040

A library composed of a complete set of fucopyranosyl-galactopyranosides was synthesized. A perbenzylated phenylthio fucopyranoside and a series of tri-O-benzyl-galactopyranosyl fluorides having single hydroxyl groups at the 2-, 3-, 4-, and 6-positions we

Full text of DOI:10.1016/j.carres.2006.03.040

Carbohydrate Research 341 (2006) 1476–1487
Synthesis of a library of fucopyranosyl-galactopyranosides
consisting of a complete set of anomeric configurations
and linkage positions
Isao Ohtsuka, Takuro Ako, Rumiko Kato, Shusaku Daikoku, Satomi Koroghi,
Takuya Kanemitsu and Osamu Kanie*
Mitsubishi Kagaku Institute of Life Sciences (MITILS), 11 Minamiooya, Machida-shi, Tokyo 194-8511, Japan
Received 27 February 2006; received in revised form 13 March 2006; accepted 27 March 2006
Available online 27 April 2006
Abstract—A library composed of a complete set of fucopyranosyl-galactopyranosides was synthesized. A perbenzylated phenylthio
fucopyranoside and a series of tri-O-benzyl-galactopyranosyl fluorides having single hydroxyl groups at the 2-, 3-, 4-, and 6-posi-
tions were used as the glycosyl donor and glycosyl acceptors, respectively. The chosen set of functionalities at the anomeric centers
enabled rapid access to the oligosaccharides based on chemoselective activation. The first coupling reaction was achieved by the
action of dimethyl(methylthio)sulfonium trifluoromethanesulfonate (DMTST). The resulting disaccharide fluoride was readily acti-
vated by hafnocene bistrifluoromethanesulfonate [Cp2Hf(OTf)2] and glycosidated with n-octanol.
ꢀ 2006 Elsevier Ltd. All rights reserved.
Keywords: Fucopyranosyl-galactopyranoside; Library; Deprotection of chloroacetyl group; Anomeric mixture; Phenylthio glycosides; Glycosyl
fluorides
1. Introduction
the exo-glycosidic atom is substituted with a sulfur or
a carbon atom.8,9 Alternatively, we propose searching
for compounds in an oligosaccharide library that mimic
a functional structure but resist hydrolysis. Before veri-
fying the hypothesis, the synthesis of a library of oligo-
saccharides must be accomplished. State-of-the-art
synthesis, however, cannot always be directly applied
to combinatorial chemistry where the synthetic protocol
should be extremely simple.7,10–14
To address the synthesis of an oligosaccharide library,
we focused on the overall efficiency of the synthetic pro-
cess. We decided to reduce (1) the number of protecting
groups and (2) the number of reactions used throughout
the synthesis. To fulfill criterion 1, benzyl groups were
selected as a sole group to protect the hydroxyl
groups.15 For criterion 2, an orthogonal glycosylation
reaction16 was used (Scheme 1). Fucose-containing oli-
gosaccharides are present in blood type antigens,17
tumor antigens,18 and are also ligands of H. pylori.19
Thus, detailed structure–activity relationships of fucosyl-
ated oligosaccharides is quite important; however, the
The research field of glycotechnology has expanded from
fundamental chemical and biological research to one of
potential pharmaceutical interest. Carbohydrates can
be considered an important class of potential pharma-
ceuticals because a wide range of functions involved in
cellular interactions are being discovered.1 The synthesis
of this class of molecules is therefore of extreme impor-
tance. Recent efforts directed at the synthesis of complex
glycoconjugates have made it possible to control the
stereo- and regiospecificity in glycosylation reactions
using solution- and solid-phase organic synthesis.2–7
Naturally-occurring oligosaccharides are susceptible
to degradation by hydrolases present in the blood-
stream. A possible way to circumvent this problem
involves the use of analogues of O-glycosides in which
*
Corresponding author. Tel.: +81 42 724 6238; fax: +81 42 724
6317; e-mail: kokanee@libra.ls.m-kagaku.co.jp
0008-6215/$ - see front matter ꢀ 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2006.03.040

I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
1477
O
SPh
(BnO)n
F
O
O
O
F
O
HO
DMTST
(BnO)n
(BnO)n
(BnO)n
O
O
O
HO
R
O
R
(BnO)n
(BnO)n
Cp2Hf(OTf)2
Scheme 1.
use of oligosaccharide libraries have not been applied to
this problem. We therefore selected the Fucp-Galp se-
quence not only to demonstrate the synthesis of a library
but also to provide a series of compounds for future bio-
logical investigations. In this paper, we report the syn-
thesis of a library of the Fucp-Galp disaccharide
consisting of all sets of anomeric configurations and
linkages.
reactions are needed and thus the above consideration
is not or primary concern, we envisioned future studies
in which longer oligosaccharides were synthesized
(Scheme 1).
We have previously reported the synthesis of tribenzyl-
ated phenylthio galactopyranosides (1–4), which carry
single hydroxyl groups at the 2-, 3-, 4-, and 6-positions,
respectively.20 To achieve orthogonal coupling, com-
pounds 1–4 have to be converted into the corresponding
fluorides (Scheme 2). First, each hydroxyl group in com-
pounds 1–4 was protected with a chloroacetyl (ClAc)
group (5–8) using chloroacetyl chloride in the presence
of pyridine. Conversion of the thioglycosides into the
glycosyl fluorides (9–12) was achieved by the action of
N,N-diethylaminosulfur trifluoride (DAST).21 We have
found that bromonium ion, usually generated from
N-bromosuccinimide (NBS), is not necessary for this
reaction. In the absence of NBS, a higher reaction
temperature (room temperature to 40 ꢁC) was required,
but the reaction was much cleaner and the desired
fluoride was obtained in almost quantitative yield. We
suggest that a Vilsmeir-type electrophilic sulfiminium
cation species is the active agent in the reaction (Scheme
3). The next step was the deprotection of the ClAc
group, which is usually achieved by treatment with
thiourea,22 1,4-diazabicyclo[2,2,2]octane (DABCO),23
2. Results and discussion
For the synthesis of the library, we planned to adopt an
orthogonal glycosylation strategy, which was considered
to be most suitable for this type of synthesis. In the syn-
thesis of the oligosaccharides iterative glycosylation
reactions and protecting group manipulations have to
be performed, and this complicates the overall synthesis
due to problems associated with the selective reaction
conditions that must be applied.4 Because the orthogo-
nal glycosylation strategy relies on only two reaction
conditions to selectively activate one of the leaving
groups over the other, the entire synthetic route becomes
simple, which makes the method advantageous over
others.16 Although in the current report we report the
synthesis of disaccharides, where only two coupling
3
OR
4
3
OR
4
OR
OR
DAST
O
O
2
2
R O
R O
SPh
CH2Cl2
F
1
1
R O
R O
R1 R2 R3 R4
R1 R2 R3 R4
9 ClAc Bn Bn Bn
10 Bn ClAc Bn Bn
11 Bn Bn ClAc Bn
12 Bn Bn Bn ClAc
1
2
3
4
H
Bn
Bn Bn
Bn Bn Bn
Bn Bn
H
H
Bn
H
Bn Bn Bn
ClAcCl
C2H4Cl2
Pyr.
N2H4•AcOH
or DABCO
5
6
7
8
ClAc Bn Bn Bn
Bn ClAc Bn Bn
Bn Bn ClAc Bn
Bn Bn Bn ClAc
13
14 Bn
15 Bn Bn
16 Bn Bn Bn
H
Bn Bn Bn
H
Bn Bn
H
Bn
H
Scheme 2.

1478
I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
-
F
F
F
Et
+
Et
F
N
S
N
Et
O
Et
PhSSF NEt
F
S
2
2
-
F
+
O
O
O
+
S
S
Ph
Ph
-
F
F
Scheme 3.
NaOCH3,24 or N2H4ÆAcOH.25 In an effort to synthesize
a series of synthetic units, we examined these reactions
and found that the reactions of N2H4ÆAcOH and DAB-
CO gave excellent results. Because the substrate for
these reactions were glycosyl fluorides, we anticipated
that the use of methoxide would generate anionic species
next to the anomeric center in the case of compound 9,
leading to poor yield due to the loss of fluorine atom.
The reaction using thiourea resulted in adequate yields
(ꢀ75%) for all substrates. DABCO was the first choice
for our substrates, except for the compound carrying a
ClAc group at the O-2 position. In the case of com-
pound 9, N2H4ÆAcOH was found to give an essentially
quantitative yield.
Having completed the syntheses of the monosaccha-
ride units, we carried out the synthesis of a disaccharide
library (Scheme 4). Glycosylation of compounds 13–16
with fucosyl donor 17 was carried out in the presence
of DMTST26,27 in dichloroethane and acetonitrile to
yield disaccharides in generally good yield according
by TLC. Because our intention was to synthesize all
possible structures, we had determined beforehand that
O
BnO
BnO
SPh
OBn
n-Octanol
Cp2Hf(OTf)2
CH2Cl2
17
DMTST
CH2Cl2
H2-Pd/C
AcOH
OBn
O
OBn
O
OBn
O
OH
O
F
F
OOct
OBn
OOct
OH
BnO
BnO
BnO
HO
BnO
BnO
BnO
HO
O
OH
O
O
O
O
O
13
OBn
BnO
OBn
BnO
OBn
HO
OH
18
22
26
OBn
O
OBn
OBn
OH
O
O
O
F
F
OBn
OOct
OBn
OOct
BnO
BnO
O
BnO
O
HO
O
HO
OBn
BnO
BnO
O
BnO
BnO
O
HO
HO
O
OH
14
OBn
OBn
OH
19
23
27
OBn
O
OBn
O
OBn
O
OH
O
O
O
O
HO
F
BnO
BnO
O
F
BnO
BnO
O
OOct
HO
HO
O
OOct
BnO
OBn
HO
OH
OBn
OBn
OBn
OBn
OH
BnO 20
24
BnO
15
28
OBn
OBn
OH
BnO
BnO
BnO
BnO
HO
HO
O
O
O
O
O
O
OH
O
O
O
O
BnO
F
BnO
F
BnO
OOct
HO
OOct
BnO
OBn
BnO
BnO
HO
OBn
OBn
OH
21
25
29
16
Scheme 4.

I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
1479
the optimal solvent system was a 1:1 mixture of dichlo-
roethane and acetonitrile, which gave anomeric mixtures
in the fucosylation reactions (data not shown). Each
anomer in the disaccharide mixtures 18–21 was not
purified at this stage to avoid unnecessary product loss,
but instead were collected as a mixture. The mixtures
were then used as the glycosyl donors and reacted with
n-octanol in the presence of Cp2Hf(OTf)2.28–30 Incorpo-
ration of an n-octyl group at the reducing anomeric cen-
ter was based on a consideration that the final products
can be distinguished easily from other hydrophilic by-
products such as hydrolysis and elimination products.31
The solvents used were dichloroethane and acetonitrile
(1:1) to obtain a ‘good’ mixture where an equal distribu-
tion of anomers was obtained. The Fucp-Galp-Octyl
disaccharides (22–25), were subjected to hydrogenation
(58%, 49%, 40%, and 68% yields for 2-, 3-, 4-, and 6-po-
sition, respectively) and finally purified by high perfor-
mance liquid chromatography (HPLC). A reverse-
phase (C18) column was successfully used to isolate all
the anomers [designated as i–iv for a/a, b/a, a/b, and
b/b Fucp/Galp, respectively] in each mixture (Table 1,
26–29). We viewed it as rather unusual that these com-
pounds could be successfully purified on reverse phase
chromatography as these compounds have the same
molecular weight. This suggests that there are differences
in the partition coefficient values (logP) between these
compounds, possible due to the packing of individual
compounds.
In conclusion, we have demonstrated the synthesis of
a complete library of Fucp-Galp-Octyl disaccharides.
The extremely short syntheses were achieved using a
semi-orthogonal glycosylation strategy. Although the
method is neither practical nor ‘efficient’ in terms of ste-
reospecific synthesis, libraries of this type may be useful
as a source of potential pharmaceutical seeds or func-
tional molecules. The NMR data of this library of com-
pounds may also be useful in the structural
characterization of newly discovered glycoconjugates.
Furthermore, structural analysis and quantitative analy-
sis of the MS/MS data obtained from the library are
currently underway32 in our laboratory using collision
induced dissociation (CID) mass spectrometry, which
will be reported elsewhere.
Table 1. Structures of the Fucp-Galp library
Linkage position Gal
Fuc
a
b
a
b
a
b
OH
OH
OH
OH
O
O
O
O
HO
OOct
OH
HO
OOct
OH
HO
HO
OOct
OH
HO
HO
OOct
OH
HO
HO
O
O
O
O
Fuc-(1!2)-Gal
Fuc-(1!3)-Gal
Fuc-(1!4)-Gal
Fuc-(1!6)-Gal
O
O
O
O
HO
OH
26-i
OH
HO
OH
26-ii
OH
HO
OH
26-iii
OH
HO
OH
26-iv
OH
O
O
O
O
HO
O
OOct
HO
O
OOct
HO
O
OOct
HO
O
OOct
HO
HO
O
HO
HO
O
HO
HO
O
HO
HO
O
OH
OH
OH
OH
OH
OH
OH
OH
27-i
27-ii
27-iii
27-iv
OH
OH
O
OOct
OH
28-iii
OH
OH
O
O
O
O
O
O
O
HO
HO
O
OOct
OH
28-ii
HO
HO
O
HO
HO
O
OOct
OH
28-i
HO
HO
O
OOct
OH
28-iv
HO
HO
OH
OH
HO
HO
OH
OH
OH
OH
OH
OH
HO
HO
HO
HO
HO
HO
HO
O
O
O
O
HO
O
O
O
O
O
O
O
O
OOct
OH
29-ii
HO
OOct
OH
29-iii
HO
OOct
OH
29-iv
HO
HO
OOct
OH
29-i
HO
HO
HO
HO

1480
I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
3. Experimental
(br d, 1H, J 2.7 Hz, H-4), 3.98 (d, 1H, J 14.8 Hz,
ClCH2CO), 3.90 (d, 1H, J 14.8 Hz, ClCH2CO), 3.64–
3.62 (m, 3H, H-5, H-6, H-60), 3.59 (dd, 1H, J3,4
2.7 Hz, H-3); 13C NMR (CDCl3): d 165.9 (ClCH2CO),
138.3–127.5 (aromatic C), 86.2 (C-1), 81.2 (C-3), 77.6
(C-5), 74.4, 73.6 (PhCH2 · 2), 72.7 (C-4), 72.0 (PhCH2),
71.4 (C-2), 68.6 (C-6), 40.8 (ClCH2CO); ESIMS: m/z
calcd for [C35H35ClO6]Na+: 641.1735. Found: 641.1732.
3.1. General methods
Analytical thin layer chromatography (TLC) was per-
formed on Merck Art. 5715, Kieselgel 60 F254/0.25 mm
thickness plates. Visualization was accomplished with
UV light and phosphomolybdic acid and/or sulfuric acid
solution followed by heating. Column chromatography
was performed with Merck Art 7734 Silica Gel 60 (70–
230 mesh). Optical rotations were measured in a
1.0 dm tube with a Horiba SEPA-200 polarimeter. Melt-
ing points were measured with Yanaco MP-S3 micro-
3.3. 3,4,6-Tri-O-benzyl-2-O-chloroacetyl-a,b-D-galacto-
pyranosyl fluoride (9)
To a solution of 5 (213.5 mg, 345 lmol) in CH2Cl2
(3.5 mL) was added DAST (85 lL, 69 lmol) at 0 ꢁC
and the mixture was stirred at 40 ꢁC for 3.5 h. The reac-
tion mixture was diluted with EtOAc and extracted with
satd aq NaHCO3 and brine. The organic layer was dried
(Na2SO4), filtered, and concentrated. The residue was
purified by chromatography on silica gel (5:1, hexane/
EtOAc) to give 9 (181.5 mg, quant., a/b = 1.00/0.32)
1
melting point apparatus. H NMR (500 MHz) spectra
were recorded with a AVANCE 500 spectrometer (Bru-
ker Biospin Inc.) in deuterated solvents using (CH3)4Si
as the internal standard. 13C NMR chemical shifts were
obtained and assigned by HSQC experiments. HPLC
was performed on Waters LC/MS System with following
setup; Pump: Waters 2525; Makeup Pump: Waters 515
HPLC Pump; Column Oven: Senshu Scientific Co. Ltd
2120; MS: Waters Micromass ZQ. High resolution mass
spectra were obtained using Q-STAR pulsar i with ESI
interface (ABS) or LCMS-IT-TOF (Shimadzu). Solvents
(HPLC): CH3CN: Merck LC/MS hyper grade; H2O:
Wako HPLC grade. Columns (HPLC): Xterra C18:
Waters Corp.; Chromolith Performance RP-18: Merck.
Doubly deionized H2O was prepared with a Milli-Q
system (Millipore Corp.). Millex-GV syringe filters
[0.22 lm · 4 mm (I.D.)] were from Nihon Millipore
Ltd. Dry solvents were used in all reactions. Solutions
were evaporated under reduced pressure at a bath
temperature not exceeding 50 ꢁC. Gel permeation chro-
matography was performed using Bio Beads SX-1
(Bio-Rad laboratories). Sep-Pak C18 was from Waters
Corp.
23
as a pale yellow syrup; ½aꢁD +44.8 (a-anomer, c 1.0,
CHCl3); Rf 0.36 (a-anomer), 0.16 (b-anomer) (5:1, hex-
1
ane/EtOAc); H NMR (CDCl3): d 7.38–7.24 (m, 30H,
2
aromatic H), 5.77 (dd, 1H, J1,2 2.7 Hz, J1,F 54.5 Hz,
3
H-1a), 5.48 (ddd, 1H, J2,3 10.0 Hz, J2,F 12.3 Hz, H-
3
2b), 5.40 (ddd, 1H, J2,3 10.4 Hz, J2,F 24.3 Hz, H-2a),
2
5.15 (dd, 1H, J1,2 7.0 Hz, J1,F 53.1 Hz, H-1b), 4.93 (d,
1H, J 11.3 Hz, PhCH2-a), 4.92 (d, 1H, J 11.5 Hz,
PhCH2-b), 4.71 (d, 1H, J 12.0 Hz, PhCH2-a), 4.67 (d,
1H, J 12.2 Hz, PhCH2-b), 4.62 (d, 1H, J 12.0 Hz,
PhCH2-a), 4.58 (d, 1H, J 11.5 Hz, PhCH2-b), 4.57 (d,
1H, J 11.3 Hz, PhCH2-a), 4.51–4.42 (m, 5H, PhCH2-
a · 2, PhCH2-b · 3), 4.12 (br t, 1H, J 6.6 Hz, H-5a),
4.07–3.92 (m, 7H, H-3a, H-4a,b, ClCH2CO-a,b), 3.71
(m, 1H, H-5b), 3.66 (m, 2H, H-6b, H-60b), 3.59 (d,
2H, J 6.5 Hz, H-6a, H-60a), 3.57 (dd, J3,4 2.4 Hz, H-
3b); 13C NMR (CDCl3): d 166.8 (ClCH2CO-a), 165.9
(ClCH2CO-b), 137.9–127.5 (aromatic C), 107.0 (d,
JC-1,F 219 Hz, C-1b), 104.5 (d, JC-1,F 226 Hz, C-1a), 78.7
3.2. Phenyl 3,4,6-tri-O-benzyl-2-O-chloroacetyl-1-thio-b-
D-galactopyranoside (5)
3
(d, JC-3,F 12 Hz, C-3b), 75.8 (C-3a), 74.9 (PhCH2-a),
3
To a solution of 133 (1.01 g, 1.86 mmol) in C2H4Cl2
(18 mL) and pyridine (1 mL) was added ClAcCl
(300 lL, 3.73 mmol) at 0 ꢁC and the mixture was stirred
at rt for 30 min. The reaction mixture was diluted with
EtOAc and extracted with N HCl, satd aq NaHCO3,
and brine. The organic layer was dried (Na2SO4), fil-
tered, and concentrated. The residue was purified by
chromatography on silica gel (5:1, hexane/EtOAc) to
give 5 (1.14 g, 99%) as needles; mp: 121.5–122.0 ꢁC;
74.6 (PhCH2-b), 74.0 (d, JC-5,F 2 Hz, C-5b), 73.6 (C-4,
2
PhCH2-b), 73.5 (PhCH2-a), 72.9 (d, JC-2,F 24 Hz, C-
2
2a), 72.7 (PhCH2-a), 72.2 (PhCH2-b), 72.1 (d, JC-2,F
3
25 Hz, C-2a), 71.9 (d, JC-5,F 3 Hz, C-5a), 68.0 (C-6b),
67.9 (C-6a), 40.6 (ClCH2CO-a,b); ESIMS: m/z calcd
for [C29H30ClFO6]Na+: 551.1607. Found: 551.1596.
3.4. 3,4,6-Tri-O-benzyl-a,b-D-galactopyranosyl fluoride
(13)
27
½aꢁD +15.9 (c 1.0, CHCl3); Rf 0.26 (5:1, hexane/EtOAc);
1H NMR (CDCl3): d 7.48–7.20 (m, 20H, aromatic H),
5.43 (t, 1H, J2,3 9.7 Hz, H-2), 4.92 (d, 1H, J 11.5 Hz,
PhCH2), 4.67 (d, 1H, J 12.2 Hz, PhCH2), 4.63 (d, 1H,
J1,2 10.0 Hz, H-1), 4.56 (d, 1H, J 11.6 Hz, PhCH2),
4.49 (d, 1H, J 11.9 Hz, PhCH2), 4.45 (d, 1H, J
12.6 Hz, PhCH2), 4.42 (d, 1H, J 11.6 Hz, PhCH2), 4.00
To a solution of 9 (46.7 mg, 88.3 lmol) in DMF
(0.88 mL) was added N2H4ÆAcOH (25 mg, 269 lmol)
at rt, and stirred overnight at rt. The reaction mixture
was diluted with EtOAc and extracted with satd aq
NaHCO3 and brine. The organic layer was dried
(Na2SO4), filtered, and concentrated to give 1334

I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
1481
(39.9 mg, quant., a/b = 1.00/0.29) as a light brown syr-
up, which was crystallized from hexane/EtOAc to give
pound 6 (55.1 mg, 89.0 lmol), CH2Cl2 (0.89 mL), and
DAST (22 lL, 18 lmol). Column chromatography
(5:1, hexane/EtOAc); yield (43.9 mg, 93%, a/b = 1.00/
26
13a as needles; mp: 60.0–61.0 ꢁC (a-anomer); ½aꢁD
27
+58.7 (a-anomer, c 1.0, CHCl3); Rf 0.24 (3:1, hexane/
EtOAc); H NMR (CDCl3): d 7.38–7.26 (m, 30H, aro-
matic H), 5.69 (dd, 1H, J1,2 2.7 Hz, J1,F 54.3 Hz, H-
1a), 5.06 (dd, 1H, J1,2 7.1 Hz, J1,F 53.7 Hz, H-1b),
0.10) as a light brown syrup; ½aꢁD +55.4 (a/b = 25/1, c
1
1.0, CHCl3); Rf 0.33 (5:1, hexane/EtOAc); NMR data
for a-anomer: 1H NMR (CDCl3): d 7.37–7.23 (m,
2
2
2
15H, aromatic H), 5.61 (dd, 1H, J1,2 2.6 Hz, J1,F
4.88 (d, 1H, J 11.3 Hz, PhCH2-a), 4.87 (d, 1H, J
11.5 Hz, PhCH2-b), 4.76–4.71 (m, 2H, PhCH2-a,b),
4.64–4.42 (m, 8H, PhCH2-a · 4, PhCH2-b · 4), 4.20
53.1 Hz, H-1), 5.29 (dd, 1H, J3,4 3.0 Hz, H-3), 4.69 (d,
1H, J 12.1 Hz, PhCH2), 4.66 (d, 1H, J 12.2 Hz, PhCH2),
4.58 (d, 1H, J 11.5 Hz, PhCH2), 4.54 (s, 1H, PhCH2),
4.52 (s, 1H, PhCH2), 4.46 (d, 1H, J 11.9 Hz, PhCH2),
4.24 (br t, 1H, J 6.8 Hz, H-5), 4.14 (br d, 1H, J
3
(br dd, 1H, J2,3 9.1 Hz, J2,F 24.8 Hz, H-2a), 4.13 (br
t, 1H, J5,6 6.6 Hz, H-5a), 4.09–4.03 (m, 2H, H-2b, H-
4a), 3.95 (br s, 1H, H-4b), 3.76 (dd, 1H, J2,3 10.1 Hz,
J 2.5 Hz, H-3a), 3.69–3.57 (m, 5H, H-5b, H-6a,b, H-
60a,b), 3.42 (dd, 1H, J2,3 9.8 Hz, J 2.4 Hz, H-3b), 2.49
(br s, 1H, OHb), 2.24 (br s, 1H, OHa); 13C NMR
(CDCl3): d 138.1–127.7 (aromatic C), 109.6 (d, JC-1,F
213 Hz, C-1b), 107.3 (d, JC-1,F 224 Hz, C-1a), 81.1 (d,
3JC-3,F 11 Hz, C-3b), 78.6 (C-3a), 74.8 (PhCH2-a), 74.6
3
2.3 Hz, H-4), 4.04 (ddd, 1H, J2,3 10.4 Hz, J2,F
24.4 Hz, H-2), 3.80 (d, 1H, J 14.8 Hz, ClCH2CO), 3.72
(d, 1H, J 14.8 Hz, ClCH2CO), 3.58 (br d, 2H, J
6.2 Hz, H-6, H-60); 13C NMR (CDCl3): d 166.5
(ClCH2CO), 137.7–127.8 (aromatic C), 105.6 (d, JC-1,
228 Hz, C-1), 75.2 (PhCH2), 74.5 (C-4), 73.5, 73.4,
F
3
73.3 (C-2, C-3, PhCH2 · 2), 70.9 (d, JC-5,F 3 Hz, C-5),
3
(PhCH2-b), 73.7 (d, JC-5,F 5 Hz, C-5b), 73.6 (PhCH2-
67.4 (C-6), 40.6 (ClCH2CO); HRTOFMS calculated
b), 73.5 (PhCH2-a), 73.0 (C-4a), 72.4 (PhCH2-b), 72.2
for [C29H30ClFO6]Na+: 551.1607. Found: 551.1598.
3
(PhCH2-a), 72.0 (C-4b), 72.0 (d, JC-5,F 3 Hz, C-5a),
2
2
71.0 (d, JC-2,F 25 Hz, C-2b), 68.4 (d, JC-2,F 12 Hz, C-
2a), 68.1 (C-6a,b); ESIMS: m/z calcd for [C27H29FO5]-
Na+: 475.1891. Found: 475.1887.
3.7. 2,4,6-Tri-O-benzyl-a,b-D-galactopyranosyl fluoride
(14)
To a solution of 10 (131 mg, 248 lmol) in EtOH/pyridine
(5:1, 2.5 mL) was added DABCO (85 mg, 743 lmol) at
room temperature, and stirred for overnight at 70 ꢁC.
The reaction mixture was diluted with EtOAc and
extracted with N HCl, satd aq NaHCO3, and brine.
The organic layer was dried over Na2SO4, filtered, and
concentrated to give 14 (110 mg, quant., a/b = 1.00/
3.5. Phenyl 2,4,6-tri-O-benzyl-3-O-chloroacetyl-1-thio-b-
D-galactopyranoside (6)
Compound 624 was prepared according to the method
described for the synthesis of compound 5 using com-
pound 224 (1.01 g, 1.86 mmol), C2H4Cl2 (18 mL), pyr-
idine (1 mL), and ClAcCl (300 lL, 3.73 mmol). Column
chromatography (5:1, hexane/EtOAc); (1.14 g, 99%) as
27
0.08) as a light brown syrup; ½aꢁD +27.5 (a/b = 25/1,
c 1.0, CHCl3); Rf 0.30 (3:1, hexane/EtOAc); NMR data
26
1
a pale yellow syrup; ½aꢁD +37.0 (c 1.0, CHCl3); Rf 0.33
for a-anomer: H NMR (CDCl3): d 7.37–7.27 (m, 15H,
2
(5:1, hexane/EtOAc); 1H NMR (CDCl3): d 7.59–7.20
(m, 20H, aromatic H), 4.98 (dd, 1H, J3,4 3.0 Hz, H-3),
4.83 (d, 1H, J 11.1 Hz, PhCH2), 4.68 (d, 1H, J1,2
9.7 Hz, H-1), 4.56 (s, 2H, PhCH2), 4.52 (d, 1H, J
11.1 Hz, PhCH2), 4.52 (d, 1H, J 11.8 Hz, PhCH2), 4.45
(d, 1H, J 11.7 Hz, PhCH2), 4.05 (br d, 1H, J 2.9 Hz,
H-4), 3.95 (t, 1H, J2,3 9.7 Hz, H-2), 3.74 (br t, 1H, J
6.6 Hz, H-5), 3.67 (br d, 2H, J 6.6 Hz, H-6, H-60), 3.60
(d, 1H, J 14.8 Hz, ClCH2CO), 3.56 (d, 1H, J 14.8 Hz,
ClCH2CO); 13C NMR (CDCl3): d 166.5 (ClCH2CO),
138.1–127.3 (aromatic C), 87.6 (C-1), 78.4 (C-3), 76.7
(C-5), 75.3 (PhCH2), 75.2 (C-2), 74.9 (PhCH2), 74.2
(C-4), 73.5 (PhCH2), 67.8 (C-6), 40.4 (ClCH2CO);
ESIMS: m/z calcd for [C35H35ClO6]Na+: 641.1735.
Found: 641.1744.
aromatic H), 5.63 (dd, 1H, J1,2 2.5 Hz, J1,F 53.7 Hz,
H-1), 4.81 (d, 1H, J 11.5 Hz, PhCH2), 4.74 (d, 1H, J
11.7 Hz, PhCH2), 4.69 (d, 1H, J 11.7 Hz, PhCH2), 4.62
(d, 1H, J 11.5 Hz, PhCH2), 4.52 (d, 1H, J 11.9 Hz,
PhCH2), 4.45 (d, 1H, J 11.9 Hz, PhCH2), 4.16 (br t,
1H, J 6.5 Hz, H-5), 4.07 (ddd, 1H, J3,4 3.3 Hz, J3,OH
4.6 Hz, H-3), 4.00 (br d, 1H, J 2.3 Hz, H-4), 3.80 (ddd,
3
1H, J2,3 10.0 Hz, J2,F 25.2 Hz, H-2), 3.61–3.55 (m, 2H,
H-6, H-60), 2.27 (d, 1H, J2,OH 5.0 Hz, OH); 13C NMR
(CDCl3): d 138.1–127.7 (aromatic C), 105.1 (d, JC-1,F
2
227 Hz, C-1), 76.5 (d, JC-2,F 24 Hz, C-2), 75.9 (C-4),
3
75.1, 73.3, 72.9 (PhCH2 · 3), 71.5 (d, JC-5,F 3 Hz, C-5),
69.7 (C-3), 68.1 (C-6); ESIMS: m/z calcd for
[C27H29FO5]Na+: 475.1891. Found: 475.1891.
3.8. Phenyl 2,3,6-tri-O-benzyl-4-O-chloroacetyl-1-thio-b-
D-galactopyranoside (7)
3.6. 2,4,6-Tri-O-benzyl-3-O-chloroacetyl-a,b-D-galacto-
pyranosyl fluoride (10)
Compound 7 was prepared according to the method
described for the synthesis of compound 5 using
compound 335 (1.02 g, 1.88 mmol), C2H4Cl2 (18 mL),
Compound 10 was prepared according to the method
described for the synthesis of compound 9 using com-

1482
I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
27
pyridine (1 mL), and ClAcCl (300 lL, 3.73 mmol).
up; ½aꢁD +14.9 (a-anomer, c 1.1, CHCl3); Rf 0.32 (3:1,
hexane/EtOAc); NMR data for a-anomer: 1H NMR
(CDCl3): d 7.36–7.26 (m, 15H, aromatic H), 5.59 (dd,
Column chromatography (5:1, hexane/EtOAc); yield
23
(1.08 g, 93.3%) as a pale yellow syrup; ½aꢁD +2.7 (c
1.0, CHCl3); Rf 0.32 (5:1, hexane/EtOAc); 1H NMR
(CDCl3): d 7.57–7.25 (m, 20H, aromatic H), 5.70 (br
d, 1H, J 2.9 Hz, H-4), 4.77 (d, 1H, J 10.9 Hz, PhCH2),
4.74 (d, 1H, J 10.2 Hz, PhCH2), 4.70 (d, 1H, J
10.2 Hz, PhCH2), 4.68 (d, 1H, J1,2 9.6 Hz, H-1), 4.56
(d, 1H, J 11.6 Hz, PhCH2), 4.49 (d, 1H, J 10.9 Hz,
PhCH2), 4.45 (d, 1H, J 11.6 Hz, PhCH2), 4.05 (d, 1H,
1H, J1,2 2.4 Hz, J1,F 52.1 Hz, H-1), 4.82 (d, 1H, J
2
11.8 Hz, PhCH2), 4.76 (d, 1H, J 11.6 Hz, PhCH2), 4.73
(d, 1H, J 11.8 Hz, PhCH2), 4.71 (d, 1H, J 11.6 Hz,
PhCH2), 4.57 (d, 1H, J 12.0 Hz, PhCH2), 4.54 (d, 1H,
J 12.0 Hz, PhCH2), 4.12 (br s, 1H, H-4), 4.08 (br t,
3
1H, J 5.7 Hz, H-5), 3.89 (ddd, 1H, J2,3 9.8 Hz, J2,F
27.5 Hz, H-2), 3.86 (br s, 1H, H-3), 3.73 (dd, 1H, J5,6
0
0
J
15.0 Hz, ClCH2CO), 3.98 (d, 1H,
J
15.0 Hz,
5.6 Hz, J6;6 10.0 Hz, H-6), 3.68 (dd, 1H, J5;6 5.9 Hz
H-60), 2.75 (br s, 1H, OH); 13C NMR (CDCl3): d
137.8–127.7 (aromatic C), 106.0 (d, JC-1,F 226 Hz, C-
ClCH2CO), 3.78 (br t, 1H, J 6.5 Hz, H-5), 3.67 (dd,
1H, J2,3 9.1 Hz, J3,4 3.2 Hz, H-3), 3.64–3.58 (m 2H, H-
2
2, H-6), 3.52 (dd, 1H, J5,6 7.4 Hz, J6;6 9.3 Hz, H-60);
1), 77.0 (C-3), 75.0 (d, JC-2,F 24 Hz, C-2), 73.6, 73.6,
0
13C NMR (CDCl3): d 166.8 (ClCH2CO), 138.0–127.6
(aromatic C), 87.7 (C-1), 81.0 (C-3), 76.6 (C-2), 75.8
(PhCH2), 75.4 (C-5), 73.7, 72.2 (PhCH2 · 2), 68.8 (C-
4), 67.5 (C-6), 40.8 (ClCH2CO); ESIMS: m/z calcd for
[C35H35ClO6]Na+: 641.1735. Found: 641.1745.
72.5 (PhCH2 · 3), 70.7 (d, JC-5,F 3 Hz, C-5), 69.0 (C-
3
6), 67.4 (C-4); ESIMS: m/z calcd for [C27H29FO5]Na+:
475.1891. Found: 475.1888.
3.11. Phenyl 2,3,4-tri-O-benzyl-6-O-chloroacetyl-1-thio-
b-D-galactopyranoside (8)
3.9. 2,3,6-Tri-O-benzyl-4-O-chloroacetyl-a,b-D-galacto-
pyranosyl fluoride (11)
Compound 8 was prepared according to the method
described for the synthesis of compound 5 using
compound 436 (1.01 g, 1.86 mmol), C2H4Cl2 (18 mL),
pyridine (1 mL), and ClAcCl (300 lL, 3.73 mmol). Col-
umn chromatography (5:1, hexane/EtOAc); yield
(1.09 g, 95%) as a pale yellow solid, which was crystal-
Compound 11 was prepared according to the method
described for the synthesis of compound 9 using com-
pound 7 (62.3 mg, 101 lmol), CH2Cl2 (1 mL), and
DAST (25 lL, 20 lmol). Column chromatography
(5:1, hexane/EtOAc); yield (47.6 mg, 89%, a/b = 1.00/
27
lized from hexane/EtOAc; mp: 72.5–73.0 ꢁC; ½aꢁD ꢂ8.3
26
1
0.03) as a pale brown syrup; ½aꢁD +17.2 (a-anomer, c
(c 1.0, CHCl3); Rf 0.19 (5:1, hexane/EtOAc); H NMR
1.0, CHCl3); Rf 0.34 (5:1, hexane/EtOAc); NMR data
for a-anomer: 1H NMR (CDCl3): d 7.37–7.29 (m,
15H, aromatic H), 5.73 (br d, 1H, J 2.5 Hz, H-4), 5.58
(CDCl3): d 7.57–7.22 (m, 20H, aromatic H), 5.00 (d,
1H, J 11.6 Hz, PhCH2), 4.83 (d, 1H, J 10.2 Hz, PhCH2),
4.81–4.75 (m, 3H, PhCH2), 4.64 (d, 1H, J 11.6 Hz,
PhCH2), 4.63 (d, 1H, J1,2 9.7 Hz, H-1), 4.36 (dd, 1H,
2
(dd, 1H, J1,2 2.6 Hz, J1,F 53.0 Hz, H-1), 4.83 (d, 1H, J
0
0
11.8 Hz, PhCH2), 4.78 (d, 1H, J 11.1 Hz, PhCH2), 4.67
(d, 1H, J 11.8 Hz, PhCH2), 4.55 (d, 1H, J 11.1 Hz,
PhCH2), 4.55 (d, 1H, J 11.9 Hz, PhCH2), 4.42 (d, 1H,
J 11.9 Hz, PhCH2), 4.26 (br t, 1H, J 6.4 Hz, H-5), 4.01
(d, 1H, J 14.8 Hz, ClCH2CO), 3.98 (dd, 1H, J3,4
3.2 Hz, H-3), 3.94 (d, 1H, J 14.9 Hz, ClCH2CO), 3.73
J5,6 7.1 Hz, J6;6 11.2 Hz, H-6), 4.13 (dd, 1H, J5;6
5.4 Hz, H-60), 3.95 (t, 1H, J2,3 9.4 Hz, H-2), 3.94 (d,
1H, J 14.7 Hz, ClCH2CO), 3.90 (d, 1H, J 15.0 Hz,
COCH2Cl), 3.83 (br d, 1H, J 2.2 Hz, H-4), 3.62–3.59
(m, 2H, H-2, H-5); 13C NMR (CDCl3): d 166.8
(ClCH2CO), 138.0–127.3 (aromatic C), 87.8 (C-1), 84.0
(C-3), 77.3 (C-2), 75.7 (PhCH2), 75.5 (C-5), 74.1, 73.2
(PhCH2 · 2), 72.9 (C-4), 64.8 (C-6), 40.6 (ClCH2CO);
ESIMS: m/z calcd for [C35H35ClO6]Na+: 641.1735.
Found: 641.1733.
3
(ddd, 1H, J2,3 10.0 Hz, J2,F 25.3 Hz, H-2), 3.53 (dd,
0
0
1H, J5,6 5,7 Hz, J6;6 9.4 Hz, H-6), 3.45 (dd, 1H, J5;6
7.3 Hz, H-6); 13C NMR (CDCl3): d 166.5 (ClCH2CO),
137.7–127.8 (aromatic C), 106.0 (d, JC-1,F 227 Hz, C-
2
1), 75.4 (C-3), 74.5 (d, JC-2,F 24 Hz, C-2), 73.9, 73.6,
3
72.2 (PhCH2 · 3), 69.5 (d, JC-5,F 3 Hz, C-5), 69.2 (C-
3.12. 2,3,4-Tri-O-benzyl-6-O-chloroacetyl-a,b-D-galacto-
pyranosyl fluoride (12)
4), 67.2 (C-6), 40.7 (ClCH2CO); ESIMS: m/z calcd for
[C29H30ClFO6]Na+: 551.1607. Found: 551.1590.
Compound 12 was prepared according to the method
described for the synthesis of compound 9 using com-
pound 8 (46.6 mg, 75.3 lmol), CH2Cl2 (0.75 mL), and
DAST (19 lL, 15 lmol). Column chromatography
(5:1, hexane/EtOAc); yield (38.0 mg, 95%, a/b = 1.00/
3.10. 2,3,6-Tri-O-benzyl-a,b-D-galactopyranosyl fluoride
(15)
Compound 15 was prepared according to the method
described for the synthesis of compound 14 using com-
pound 11 (73.6 mg, 139 lmol), EtOH/pyridine (5:1,
1.4 mL), and DABCO (48 mg, 42 lmol); yield
(63.0 mg, quant., a/b = 1.00/0.05) as a pale yellow syr-
25
0.01) as a pale brown syrup; ½aꢁD ꢂ2.6 (a/b = 12/1, c
1.1, CHCl3); Rf 0.25 (5:1, hexane/EtOAc); NMR data
for a-anomer: 1H NMR (CDCl3): d 7.41–7.28 (m,
2
15H, aromatic H), 5.58 (dd, 1H, J1,2 2.6 Hz, J1,F

I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
1483
53.5 Hz, H-1), 4.98 (d, 1H, J 11.4 Hz, PhCH2), 4.89 (d,
1H, J 11.7 Hz, PhCH2), 4.86 (d, 1H, J 11.8 Hz, PhCH2),
4.79 (d, 1H, J 11.7 Hz, PhCH2), 4.73 (d, 1H, J 11.8 Hz,
PhCH2), 4.62 (d, 1H, J 11.4 Hz, PhCH2), 4.25 (dd, 1H,
mixture of fluorides showed anomeric protons around
5.1–5.7 ppm with typical coupling constants (J1,2 ꢀ 3.5
and ꢀ7.8 Hz) corresponding to the a- and b-anomers
and those of J1,F (ꢀ53 Hz).
0
J5,6 6.5 Hz, J6;6 10.8 Hz, H-6), 4.13–4.02 (m, 3H, H-2,
H-5, H-60), 3.98–3.91 (m, 3H, H-3, ClCH2CO), 3.91
(br s, 1H, H-4); 13C NMR (CDCl3): d 166.8 (ClCH2CO),
138.1–127.6 (aromatic C), 106.0 (d, JC-1,F 227 Hz, C-1),
3.14.2. Glycosylation of n-octanol reaction with disac-
charide fluoride. Mixture A: Compound 18 (34 mg,
39.2 lmol) and n-octanol (12.3 lL, 78.3 lmol) dissolved
in CH3CN (1.0 mL) was stirred in the presence of
AW300 molecular sieves (500 mg) for 2 h under a N2
atmosphere. Mixture B: To a mixture of AgOTf
(30.0 mg, 118 lmol) and AW300 molecular sieves
(500 mg) in C2H4Cl2 (1.0 mL), which was stirred for
2 h under a N2 atmosphere, was added Cp2HfCl2
(22.2 mg, 58.8 lmol), and the mixture was further stir-
red for 2 h. The mixture was then centrifuged at 2000
rpm for 10 min to remove precipitated AgCl. To mixture
A, a supernatant of a mixture B was added at ꢂ20 ꢁC,
and resulting mixture was stirred for 4 h. The reaction
was terminated by adding Et3N, the mixture was filtered
through Celite pad and washed with CH2Cl2. The solu-
tion was washed with satd aq NaHCO3 and the organic
layer was dried over MgSO4 and concentrated. The res-
idue was purified on a column of Bio Beads SX-1 (tolu-
ene) to afford 22, 55 mg, as an anomeric mixture.
2
78.2 (C-3), 75.6 (d, JC-2,F 24 Hz, C-2), 74.5 (PhCH2),
3
73.8 (PhCH2), 73.7 (C-4), 73.5 (PhCH2), 70.6 (d, JC-
3 Hz, C-5), 64.5 (C-6), 40.6 (ClCH2CO); ESIMS:
5,F
m/z calcd for [C29H30ClFO6]Na+: 551.1607. Found:
551.1600.
3.13. 2,3,6-Tri-O-benzyl-a,b-D-galactopyranosyl fluoride
(16)
Compound 16 was prepared according to the method
described for the synthesis of compound 14 using com-
pound 12 (71.2 mg, 135 lmol), EtOH/pyridine (5:1,
1.4 mL), and DABCO (46.0 mg, 402 lmol); yield
(60.9 mg, quant., a/b = 1.00/<0.01) as a pale yellow syr-
26
up; ½aꢁD +2.0 (a/b = 25/1, c 1.2, CHCl3); Rf 0.27 (2:1,
hexane/EtOAc); NMR data for a-anomer: 1H NMR
(CDCl3): d 7.40–7.26 (m, 15H, aromatic H), 5.61 (dd,
2
1H, J1,2 2.6 Hz, J1,F 53.9 Hz, H-1), 4.97 (d, 1H, J
11.5 Hz, PhCH2), 4.86 (d, 2H, J 11.7 Hz, PhCH2), 4.77
(d, 1H, J 11.8 Hz, PhCH2), 4.73 (d, 1H, J 11.8 Hz,
PhCH2), 4.64 (d, 1H, J 11.5 Hz, PhCH2), 4.06 (ddd,
3.14.3. Hydrogenation of disaccharide. Compound 22
(55 mg, mixture) dissolved in AcOH (4.0 mL) was stir-
red in the presence of 5% Pd/C (60 mg) for 12 h under
a H2 atmosphere. The mixture was filtered through Cel-
ite pad and washed with CH3OH. After concentration,
the residue was purified on a Sep-Pak C18 (elution with
H2O and then CH3OH). The latter solution, containing
the desired octyl glycoside of the disaccharide, was con-
centrated. The residue was dissolved in H2O, filtered
through Millex GV filter, and concentrated to give com-
pound 26 (11 mg, 25.1 lmol; 58%; three steps). Ratio:
26-i:26-ii:26-iii:26-iv 12:1:37:21 (estimated by LC–MS,
based on assumption that these compounds are similarly
ionized).
3
1H, J2,3 9.8 Hz, J2,F 25.2 Hz, H-2), 3.96–3.91 (m, 3H,
0
H-3, H-4, H-5), 3.73 (dd, 1H, J5,6 6.3 Hz, J6;6 11.3 Hz,
H-6), 3.53 (dd, 1H, J5;6 5.6 Hz, H-60); 13C NMR
0
(CDCl3): d 138.2–127.5 (aromatic C), 106.1 (d, JC-1,F
2
226 Hz, C-1), 78.4 (C-3), 75.7 (d, JC-2,F 24 Hz, C-2),
74.5 (PhCH2), 74.2 (C-4), 73.7, 73.3 (PhCH2 · 2), 73.0
3
(d, JC-5,F 3 Hz, C-5), 61.8 (C-6); ESIMS: m/z calcd
for [C27H29FO5]Na+: 475.1891. Found: 475.1902.
3.14. Synthesis of octyl L-fucopyranosyl-(1!2)-D-galacto-
pyranosides (26)
3.14.1. Glycosylation of 3,4,6-tri-O-benzyl galacto-
pyranosyl fluoride with phenyl 2,3,4-tri-O-benzyl-1-thio-
fucopyranoside. A mixture of compound 17 (58 mg,
0.11 mmol), 13 (50 mg, 0.11 mmol), and AW300 mole-
cular sieves (500 mg) in C2H4Cl2 (1.0 mL) and CH3CN
(1.0 mL) was stirred for 3 h under an N2 atmosphere.
To this mixture, MeSSMe (19.9 lL, 0.22 mmol) and
MeOTf (25.0 lL, 0.22 mmol) were added at 0 ꢁC and
the mixture was stirred for further 4 h. Et3N was added
to the reaction mixture, the mixture was filtered through
Celite pad and washed with CH2Cl2, which was washed
with satd aq NaHCO3. The organic layer was dried
(MgSO4) and concentrated to dryness. The residue was
purified on silica gel column chromatography (15:1, tol-
uene/EtOAc) to yield 18 (86 mg, 0.99 mmol; 90%). The
3.14.4. Isolation of isomers using HPLC. Compound
26 was purified using HPLC–MS. Flow rate: 1.0 mL/
min; column: Cadenza CD-C18, 3 lm, 75 mm (L) ·
4.6 mm (I.D.); mobile phase: CH3CN/H2O (17:83);
column temperature: 15 ꢁC; detection (m/z): 461.3
[M+Na]+; retention times (min): 46.4 (26-ii; b-Fuc-a-
Gal), 49.0 (26-iv; b-Fuc-b-Gal), 73.3 (26-i; a-Fuc-a-
Gal), and 93.3 (26-iii; a-Fuc-b-Gal).
3.14.4.1. Octyl a-L-fucopyranosyl-(1!2)-a-D-galacto-
pyranoside (26-i). 1H NMR (D2O): d 5.05 (d, 1H, J1 ;2
0
0
3.6 Hz, H-10), 5.02 (d, 1H, J1,2 3.5 Hz, H-1), 4.09
(q, 1H, J5 ;6 6.5 Hz, H-50), 4.00 (br d, 1H, H-4), 3.97
(dd, 1H, H-3), 3.91–3.85 (m, 2H, H-30, H-5), 3.81–3.74
(m, 4H, H-2, H-20, H-40, OCH2CH2(CH2)5CH3), 3.72
0
0

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I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
(d, 2H, J5,6a, J5,6b 5.8 Hz, H-6a, H-6b), 3.48 (q, 1H,
OCH2CH2(CH2)5CH3), 1.62 (m, 2H, OCH2CH2(CH2)5-
CH3), 1.38–1.21 (m, 13H, H-60,OCH2CH2(CH2)5CH3),
0.85 (t, 3H, OCH2CH2(CH2)5CH3); 13C NMR (D2O): d
102.0 (C-10), 98.3 (C-1), 78.3 (C-2), 72.2 (C-40), 71.2
(C-5), 69.8 (C-4), 69.7 (C-30), 69.0 (C-3), 69.0 (C-20),
68.8 (OCH2CH2(CH2)5CH3), 67.8 (C-50), 61.5 (C-6),
31.4 (OCH2CH2(CH2)5CH3), 29.0 (OCH2CH2(CH2)5-
CH3), 28.9 (OCH2CH2(CH2)5CH3), 25.8 (OCH2-
CH2(CH2)5CH3), 22.5 (OCH2CH2(CH2)5CH3), 16.1
(C-60), 13.9 (OCH2CH2(CH2)5CH3); ESIMS m/z calcd
for [C20H38O10]Na+: 461.2357. Found: 461.2353.
(m, 2H, H-4, OCH2CH2(CH2)5CH3), 3.79–3.72 (m,
4H, H-40, H-50, H-6a, H-6b), 3.71–3.61 (m, 5H, H-2,
H-3, H-30, H-5, OCH2CH2(CH2)5CH3), 3.48 (t, 1H,
J2 ;3 8.4 Hz, H-20), 1.62 (m, 2H, OCH2CH2(CH2)5CH3),
1.35–1.24 (m, 13H, H-60, OCH2CH2(CH2)5CH3), 0.84
(t, 3H, OCH2CH2(CH2)5CH3); 13C NMR (D2O): d
106.4 (C-1), 102.4 (C-10), 77.4 (C-2), 75.2 (C-5), 73.3
(C-30), 72.1 (C-3), 71.7 (C-40), 71.2 (C-50), 71.2
(OCH2CH2(CH2)5CH3), 71.2 (C-20), 68.6 (C-4), 61.2
(C-6), 31.1 (OCH2CH2(CH2)5CH3), 29.1 (OCH2CH2-
(CH2)5CH3), 28.6 (OCH2CH2(CH2)5CH3), 25.6 (OCH2-
CH2(CH2)5CH3), 22.2 (OCH2CH2(CH2)5CH3), 15.8
(C-60), 13.9 (OCH2CH2(CH2)5CH3); ESIMS m/z calcd
for [C20H38O10]Na+: 461.2357. Found: 461.2354.
0
0
3.14.4.2. Octyl b-L-fucopyranosyl-(1!2)-a-D-galacto-
pyranoside (26-ii). 1H NMR (D2O): d 5.08 (d, 1H, J1,2
3.7 Hz, H-1), 4.45 (d, 1H, J1 ;2 7.9 Hz, H-10), 4.01 (br d,
1H, J3,4 2.8 Hz, H-4), 3.98 (dd, 1H, J2,3 7.0 Hz, H-2),
3.15. Synthesis of octyl L-fucopyranosyl-(1!3)-D-galac-
0
0
topyranosides (27)
0
0
3.93–3.88 (m, 2H, H-3, H-5), 3.77 (q, 1H, J5 ;6 6.1 Hz,
H-50), 3.74–3.66 (m, 4H, H-40, H-6a, H-6b,
The title compound was synthesized according to the
procedure described for the synthesis of compound 26.
Overall yield: 49% (three steps after Sep-Pak). Ratio:
27-i:27-ii:27-iii:27-iv 1.0:1.0:1.7:1.6 (estimated by LC–
MS based on assumption that these compounds are sim-
ilarly ionized). HPLC conditions follows; flow rate:
1.0 mL/min; column: [Chromolith Performance RP-
18e, 200 mm (L) · 4.6 mm (I.D.) (two columns were
connected)]; mobile phase: CH3CN/H2O 17:83; column
temperature: 30 ꢁC; detection (m/z): 461.3 [M+Na]+;
retention times (min): 74.2 (27-iii; a-Fuc-b-Gal), 87.4
(27-i; a-Fuc-a-Gal), 123 (27-iv; b-Fuc-b-Gal), and 140
(27-ii; b-Fuc-a-Gal).
0
0
OCH2CH2(CH2)5CH3), 3.61 (dd, 1H, J3 ;4 3.0 Hz, H-
30), 3.53 (m, 1H, OCH2CH2(CH2)5CH3), 3.49 (dd, 1H,
J2 ,3 9.8 Hz, H-20), 1.59 (m, 2H, OCH2CH2(CH2)5CH3),
1.34–1.21 (m, 13H, H-60, OCH2CH2(CH2)5CH3), 0.83
(t, 3H, OCH2CH2(CH2)5CH3); 13C NMR (D2O): d
101.5 (C-10), 96.0 (C-1), 75.6 (C-2), 72.7 (C-30), 71.1
(C-40), 71.0 (C-50), 70.7 (C-3), 70.5 (C-20), 68.9 (C-4),
68.3 (OCH2CH2(CH2)5CH3), 68.0 (C-5), 61.0 (C-6),
31.1 (OCH2CH2(CH2)5CH3), 28.7 (OCH2CH2(CH2)5-
CH3), 28.6 (OCH2CH2(CH2)5CH3), 25.8 (OCH2-
CH2(CH2)5CH3), 22.2 (OCH2CH2(CH2)5CH3), 15.4
(C-60), 13.6 (OCH2CH2(CH2)5CH3); ESIMS m/z calcd
for [C20H38O10]Na+: 461.2357. Found: 461.2354.
0
0
3.15.1. Octyl a-L-fucopyranosyl-(1!3)-a-D-galacto-
pyranoside (27-i). 1H NMR (D2O): d 5.13 (d, 1H,
3.14.4.3. Octyl a-L-fucopyranosyl-(1!2)-b-D-galacto-
pyranoside (26-iii).37 1H NMR (D2O): d 5.26 (d, 1H,
J1 ;2 4.0 Hz, H-10), 4.94 (d, 1H, J1,2 3.7 Hz, H-1), 4.17
0
0
J1 ;2 3.6 Hz, H-10), 4.62 (d, 1H, J1,2 7.9 Hz, H-1), 4.34
(q, 1H, J5 ;6 6.7 Hz, H-50), 4.05 (br d, 1H, J3,4 2.9 Hz,
0
0
0
0
(q, 1H, J5 ;6 6.4 Hz, H-50), 3.93–3.88 (m, 2H, H-4,
OCH2CH2(CH2)5CH3), 3.85–3.82 (m, 2H, H-3, H-30),
3.78–3.71 (m, 4H, H-20, H-40, H-6a, H-6b), 3.66–3.63
(m, 2H, H-5, OCH2CH2(CH2)5CH3), 3.57 (t, 1H, J2,3
8.6 Hz, H-2), 1.60 (m, 2H, OCH2CH2(CH2)5CH3),
1.30–1.26 (m, 10H, OCH2CH2(CH2)5CH3), 1.20 (d,
H-4), 3.99–3.91 (m, 3H, H-2, H-30, H-5), 3.86 (dd, 1H,
0
0
0
0
J2,3 10.6 Hz, J3,4 2.9 Hz, H-3), 3.81 (br d, 1H, J3 ;4
2.7 Hz, H-40), 3.77 (dd, 1H, J2 ;3 10.4 Hz, H-20), 3.74–
3.70 (m, 3H, H-6a, H-6b, OCH2CH2(CH2)5CH3), 3.52
(m, 1H, OCH2CH2(CH2)5CH3), 1.61 (m, 2H, OCH2-
CH2(CH2)5CH3), 1.36–1.26 (m, 10H, OCH2CH2-
0
0
3H, J5 ;6 6.6 Hz, H-60), 0.85 (t, 3H, OCH2CH2-
(CH2)5CH3); 13C NMR (D2O): d 102.0 (C-1), 99.6
(C-10), 76.7 (C-2), 75.3 (C-5), 74.2 (C-3), 72.3 (C-20),
71.0 (OCH2CH2(CH2)5CH3), 70.0 (C-30), 69.3 (C-4),
68.7 (C-40), 67.2 (C-50), 61.3 (C-6), 31.4 (OCH2-
CH2(CH2)5CH3), 29.3 (OCH2CH2(CH2)5CH3), 28.9
(OCH2CH2(CH2)5CH3), 25.7 (OCH2CH2(CH2)5CH3),
22.4 (OCH2CH2(CH2)5CH3), 15.7 (C-60), 13.9 (OCH2-
CH2(CH2)5CH3); ESIMS m/z calcd for [C20H38O10]-
Na+: 461.2357. Found: 461.2355.
(CH2)5CH3), 1.19 (d, 3H, J5 ;6 6.6 Hz, H-60), 0.84 (t,
3H, OCH2CH2(CH2)5CH3); 13C NMR (D2O): d 101.4
(C-10), 98.6 (C-1), 78.5 (C-3), 72.4 (C-40), 71.3 (C-5),
69.8 (C-30), 69.8 (C-4), 69.0 (C-20), 68.8 (OCH2-
CH2(CH2)5CH3), 67.8 (C-2), 67.5 (C-50), 61.5 (C-6),
31.4 (OCH2CH2(CH2)5CH3), 28.7 (OCH2CH2(CH2)5-
CH3), 28.6 (OCH2CH2(CH2)5CH3), 25.8 (OCH2-
CH2(CH2)5CH3), 22.2 (OCH2CH2(CH2)5CH3), 15.8
(C-60), 13.9 (OCH2CH2(CH2)5CH3); ESIMS m/z calcd
for [C20H38O10]Na+: 461.2357. Found: 461.2350.
0
0
0
0
3.14.4.4. Octyl b-L-fucopyranosyl-(1!2)-b-D-galacto-
3.15.2. Octyl b-L-fucopyranosyl-(1!3)-a-D-galacto-
pyranoside (27-ii). 1H NMR (D2O): d 4.97 (d, 1H,
pyranoside (26-iv). 1H NMR (D2O): d 4.67 (d, 1H, J1 ;2
0
0
7.8 Hz, H-10), 4.54 (m, 1H, J1,2 7.3 Hz, H-1), 3.95–3.91
J1,2 3.8 Hz, H-1), 4.67 (d, 1H, J1 ;2 7.8 Hz, H-10), 4.18
0
0

I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
1485
(br d, 1H, J3,4 2.7 Hz, H-4), 4.02 (dd, 1H, J2,3 10.1 Hz,
J3,4 2.7 Hz, H-3), 3.92 (m, 2H, H-2, H-5), 3.78 (q, 1H,
3.16. Synthesis of octyl L-fucopyranosyl-(1!4)-D-galac-
topyranosides (28)
J5 ;6 6.1 Hz, H-50), 3.74–3.69 (m, 4H, H-40, H-6a, H-
6b, OCH2CH2(CH2)5CH3), 3.65 (dd, 1H, J2,3 9.9 Hz,
J3,4 3.4 Hz, H-30), 3.52 (m, 2H, OCH2CH2(CH2)5CH3),
1.62 (m, 2H, OCH2CH2(CH2)5CH3), 1.37–1.24 (m,
13H, H-60,OCH2CH2(CH2)5CH3), 0.85 (t, 3H, OCH2-
0
0
The title compound was synthesized according to the
procedure described for the synthesis of compound 26.
Overall yield: 40% (three steps after Sep-Pak). Ratio:
28-i:28-ii:28-iii:28-iv 1.0:3.1:7.8:4.2 (estimated by LC–
MS based on assumption that these compounds are sim-
ilarly ionized). HPLC conditions follows; flow rate:
1.0 mL/min; column: [Chromolith Performance RP-
18e, 200 mm (L) · 4.6 mm (I.D.) (two columns were
connected)]; mobile phase: CH3CN/H2O 20:80; column
temperature: 30 ꢁC; detection (m/z): 461.3 [M+Na]+;
retention times (min): 36.9 (28-iv; b-Fuc-b-Gal), 49.8
(28-ii; b-Fuc-a-Gal), 54.4 (28-iii; a-Fuc-b-Gal), and
62.4 (28-i; a-Fuc-a-Gal).
13
CH2(CH2)5CH3); C NMR (D2O): d 100.9 (C-10),
98.3 (C-1), 77.7 (C-3), 73.3 (C-30), 71.8 (C-40), 71.4
(C-50), 71.0 (C-5), 70.9 (C-20), 68.8 (OCH2CH2(CH2)5-
CH3), 67.2 (C-2), 67.2 (C-4), 61.4 (C-6), 31.4 (OCH2-
CH2(CH2)5CH3), 28.8 (OCH2CH2(CH2)5CH3), 28.7
(OCH2CH2(CH2)5CH3), 25.7 (OCH2CH2(CH2)5CH3),
22.9 (OCH2CH2(CH2)5CH3), 15.7 (C-60), 13.9 (OCH2-
CH2(CH2)5CH3); ESIMS m/z calcd for [C20H38O10]-
Na+: 461.2357. Found: 461.2358.
3.15.3. Octyl a-L-fucopyranosyl-(1!3)-b-D-galacto-
pyranoside (27-iii). 1H NMR (D2O): d 5.15 (d, 1H,
3.16.1. Octyl a-L-fucopyranosyl-(1!4)-a-D-galactopy-
ranoside (28-i). 1H NMR (D2O): d 5.12 (d, 1H, J1 ;2
0
0
J1 ;2 3.9 Hz, H-10), 4.43 (m, 1H, J1,2 7.4 Hz, H-1), 4.18
4.0 Hz, H-10), 4.94 (d, 1H, J1,2 3.1 Hz, H-1), 4.13 (q,
1H, H-50), 4.04 (br d, 1H, H-4), 3.99 (dd, 1H, J5,6a
4.2 Hz, J5,6b 7.6 Hz, H-5), 3.89 (m, 3H, H-2, H-3,
H-30), 3.82 (m, 2H, H-20, H-40), 3.76–3.68 (m, 3H,
H-6a, H-6b, OCH2CH2(CH2)5CH3), 3.53 (m, 1H,
OCH2CH2(CH2)5CH3), 1.62 (m, 2H, OCH2CH2(CH2)5-
CH3), 1.39–1.23 (m, 10H, OCH2CH2(CH2)5CH3), 1.22
0
0
(q, 1H, J5 ;6 6.6 Hz, H-50), 4.00 (br d, 1H, H-4), 3.94–
0
0
3.89 (m, 2H, H-30, OCH2CH2(CH2)5CH3), 3.81 (br d,
1H, J3 ;4 3.0 Hz, H-40), 3.78–3.62 (m, 7H, H-2, H-20,
H-3, H-5, H-6a, H-6b, OCH2CH2(CH2)5CH3), 1.61
(m, 2H, OCH2CH2(CH2)5CH3), 1.34–1.26 (m, 10H,
0
0
0
0
OCH2CH2(CH2)5CH3), 1.19 (d, 3H, J5 ;6 6.6 Hz,
H-60), 0.84 (t, 3H, OCH2CH2(CH2)5CH3); 13C NMR
(D2O): d 102.8 (C-1), 101.3 (C-10), 81.3 (C-3), 75.4
(C-5), 72.2 (C-40), 71.0 (OCH2CH2(CH2)5CH3), 70.5
(C-2), 69.8 (C-30), 69.0 (C-4), 68.8 (C-20), 67.6
(C-50), 61.2 (C-6), 31.1 (OCH2CH2(CH2)5CH3), 28.9
(OCH2CH2(CH2)5CH3), 28.6 (OCH2CH2(CH2)5CH3),
(d, 3H, J5 ;6 6.5 Hz, H-60), 0.85 (t, 3H, OCH2-
0
0
13
CH2(CH2)5CH3); C NMR (D2O): d 101.8 (C-10),
98.6 (C-1), 79.8 (C-4), 72.1 (C-40), 71.4 (C-5), 70.7 (C-
3), 70.0 (C-6), 69.7 (C-30), 69.5 (C-2), 69.3 (C-20), 68.8
(OCH2CH2(CH2)5CH3), 67.9 (C-50), 31.1 (OCH2-
CH2(CH2)5CH3), 28.7 (OCH2CH2(CH2)5CH3), 28.7
(OCH2CH2(CH2)5CH3), 25.6 (OCH2CH2(CH2)5CH3),
22.1 (OCH2CH2(CH2)5CH3), 15.7 (C-60), 12.4 (OCH2-
CH2(CH2)5CH3); ESIMS m/z calcd for [C20H38-
O10]Na+: 461.2357. Found: 461.2355.
25.4
(OCH2CH2(CH2)5CH3),
22.2
(OCH2CH2-
(CH2)5CH3), 15.7 (C-60), 13.9 (OCH2CH2(CH2)5CH3);
ESIMS m/z calcd for [C20H38O10]Na+: 461.2357.
Found: 461.2349.
3.15.4. Octyl b-L-fucopyranosyl-(1!3)-b-D-galactopy-
3.16.2. Octyl b-L-fucopyranosyl-(1!4)-a-D-galacto-
pyranoside (28-ii). 1H NMR (D2O): d 4.95 (d, 1H,
ranoside (27-iv). 1H NMR (D2O): d 4.49 (d, 1H, J1 ;2
0
0
7.8 Hz, H-10), 4.42 (d, 1H, J1,2 8.0 Hz, H-1), 4.12 (br
d, 1H, J3,4 3.2 Hz, H-4), 3.91 (q, 1H, OCH2CH2-
(CH2)5CH3), 3.84 (dd, 1H, J2,3 9.0 Hz, H-3), 3.80–3.72
(m, 4H, H-40, H-50, H-6a, H-6b), 3.69–3.64 (m, 3H,
H-30, H-5, OCH2CH2(CH2)5CH3), 3.59 (t, 1H, H-2),
J1,2 3.2 Hz, H-1), 4.33 (d, 1H, J1 ;2 7.8 Hz, H-10), 4.15
(br d, 1H, H-4), 4.02 (t, 1H, J5,6a, J5,6b 6.2 Hz, H-5),
3.84–3.78 (m, 5H, H-2, H-3, H-50, H-6a, H-6b), 3.74–
3.69 (m, 2H, H-40, OCH2CH2(CH2)5CH3), 3.64 (dd,
0
0
1H, J3 ;2 10.0 Hz, J3 ;4 3.8 Hz, H-30), 3.52 (m, 2H,
H-20, OCH2CH2(CH2)5CH3), 1.61 (m, 2H, OCH2CH2-
(CH2)5CH3), 1.34–1.23 (m, 13H, H-60, OCH2CH2-
(CH2)5CH3), 0.85 (t, 3H, OCH2CH2(CH2)5CH3); 13C
NMR (D2O): d 103.5 (C-10), 98.9 (C-1), 79.5 (C-4),
73.1 (C-30), 71.8 (C-40), 71.4 (C-5), 71.4 (C-50), 71.1
(C-20), 69.5 (C-2), 69.5 (C-3), 69.1 (OCH2CH2(CH2)5-
CH3), 61.1 (C-6), 31.4 (OCH2CH2(CH2)5CH3), 28.6
(OCH2CH2(CH2)5CH3), 28.6 (OCH2CH2(CH2)5CH3),
25.6 (OCH2CH2(CH2)5CH3), 22.7 (OCH2CH2(CH2)5-
CH3), 15.8 (C-60), 12.4 (OCH2CH2(CH2)5CH3); ESIMS
m/z calcd for [C20H38O10]Na+: 461.2357. Found:
461.2348.
0
0
0
0
3.52 (t, 1H, J2 ;3 8.8 Hz, H-20), 1.61 (m, 2H, OCH2-
CH2(CH2)5CH3), 1.35–1.24 (m, 13H, H-60,OCH2-
CH2(CH2)5CH3), 0.84 (t, 3H, OCH2CH2(CH2)5CH3);
13C NMR (D2O): d 102.9 (C-1), 101.0 (C-10), 80.5
(C-3), 75.2 (C-30), 73.3 (C-5), 71.7 (C-40), 71.6 (C-50),
71.0 (OCH2CH2(CH2)5CH3), 70.9 (C-20), 69.8 (C-2),
66.6 (C-4), 61.2 (C-6), 31.4 (OCH2CH2(CH2)5CH3),
28.9 (OCH2CH2(CH2)5CH3), 28.6 (OCH2CH2(CH2)5-
CH3), 25.4 (OCH2CH2(CH2)5CH3), 22.2 (OCH2CH2-
(CH2)5CH3), 15.9 (C-60), 13.9 (OCH2CH2(CH2)5CH3);
ESIMS m/z calcd for [C20H38O10]Na+: 461.2357.
Found: 461.2362.
0
0

1486
I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
3.16.3. Octyl a-L-fucopyranosyl-(1!4)-b-D-galacto-
pyranoside (28-iii). 1H NMR (D2O): d 5.14 (d, 1H,
3.17.1. Octyl a-L-fucopyranosyl-(1!6)-a-D-galacto-
pyranoside (29-i). 1H NMR (D2O): d 4.94 (d, 1H, J1 ;2
0
0
J1 ;2 3.9 Hz, H-10), 4.41 (d, 1H, J1,2 0.9 Hz, H-1), 4.15
3.3 Hz, H-10), 4.90 (d, 1H, J1,2 3.6 Hz, H-1), 4.10 (m, 1H,
0
0
(q, 1H, J5 ;6 6.7 Hz, H-50), 4.00 (br d, 1H, J3,4 2.9 Hz,
H-4), 3.93–3.88 (m, 2H, H-30, OCH2CH2(CH2)5CH3),
3.82 (m, 2H, H-20, H-40), 3.75–3.63 (m, 5H, H-3, H-5,
H-6a, H-6b, OCH2CH2(CH2)5CH3), 3.55 (dd, 1H, J2,3
9.5 Hz, H-2), 1.61 (m, 2H, OCH2CH2(CH2)5CH3),
1.36–1.21 (m, 10H, OCH2CH2(CH2)5CH3), 1.21
H-5), 4.05 (q, 1H, J5 ;6 6.9 Hz, H-50), 3.97 (br d, 1H, J3,4
1.8 Hz, H-4), 3.85–3.75 (m, 6H, H-2, H-20, H-3, H-30,
H-4, H-6a), 3.72–3.61 (m, 2H, H-6b, OCH2CH2(CH2)5-
CH3), 3.54 (m, 1H, OCH2CH2(CH2)5CH3), 1.62 (m, 2H,
OCH2CH2(CH2)5CH3), 1.37–1.23 (m, 10H, OCH2CH2-
0
0
0
0
(CH2)5CH3), 1.21 (d, 3H, J5 ;6 6.6 Hz, H-60), 0.85 (t, 3H,
OCH2CH2(CH2)5CH3); 13C NMR (D2O): d 98.9 (C-1),
98.8 (C-10), 72.2 (C-40), 70.0 (C-5), 69.7 (C-4), 69.7 (C-3),
69.7 (C-30), 68.9 (OCH2CH2(CH2)5CH3), 68.5 (C-2),
68.5 (C-20), 67.2 (C-6), 67.2 (C-50), 31.4 (OCH2CH2-
(CH2)5CH3), 28.9 (OCH2CH2(CH2)5CH3), 28.6 (OCH2-
CH2(CH2)5CH3), 25.7 (OCH2CH2(CH2)5CH3), 22.1
(OCH2CH2(CH2)5CH3), 15.6 (C-60), 12.4 (OCH2CH2-
(CH2)5CH3); ESIMS m/z calcd for [C20H38O10]Na+:
461.2357. Found: 461.2356.
0
0
(d, 3H, J5 ;6 6.6 Hz, H-60), 0.85 (t, 3H, OCH2-
CH2(CH2)5CH3); 13C NMR (D2O): d 102.9 (C-1),
101.4 (C-10), 78.7 (C-4), 75.4 (C-5), 74.2 (C-3), 72.1
(C-40), 71.8 (C-2), 71.1 (OCH2CH2(CH2)5CH3), 70.0
(C-30), 69.3 (C-20), 67.9 (C-50), 61.5 (C-6), 31.4
(OCH2CH2(CH2)5CH3), 28.9 (OCH2CH2(CH2)5CH3),
28.6 (OCH2CH2(CH2)5CH3), 25.2 (OCH2CH2(CH2)5-
CH3), 22.9 (OCH2CH2(CH2)5CH3), 15.7 (C-60), 13.4
(OCH2CH2(CH2)5CH3); ESIMS m/z calcd for
[C20H38O10]Na+: 461.2357. Found: 461.2352.
0
0
3.17.2. Octyl b-L-fucopyranosyl-(1!6)-a-D-galacto-
pyranoside (29-ii). 1H NMR (D2O): d 4.91 (d, 1H,
3.16.4. Octyl b-L-fucopyranosyl-(1!4)-b-D-galacto-
pyranoside (28-iv). 1H NMR (D2O): d 4.41 (d, 1H,
J1,2 3.7 Hz, H-1), 4.38 (d, 1H, J1 ;2 7.9 Hz, H-10), 4.10
(septet, 1H, H-5), 4.02 (br d, 1H, J3,4 3.0 Hz, H-4),
3.95 (dd, 1H, J5,6a 6.8 Hz, J6a,6b 10.8 Hz, H-6a), 3.85–
3.69 (m, 6H, H-2, H-3, H-40, H-50, H-6b, OCH2-
0
0
J1,2 7.9 Hz, H-1), 4.31 (d, 1H, J1 ;2 7.7 Hz, H-10), 4.10
(br d, 1H, J3,4 3.1Hz, H-4), 3.89 (ddd, 1H, OCH2-
CH2(CH2)5CH3), 3.85 (dd, 1H, J6a,5 7.7 Hz, J6a,6b
11.5 Hz, H-6a), 3.81–3.73 (m, 3H, H-40, H-5, H-50),
3.69–3.59 (m, 3H, H-3, H-30, OCH2CH2(CH2)5CH3),
0
0
0
0
0
0
CH2(CH2)5CH3), 3.62 (dd, 1H, J2 ;3 9.9 Hz, J3 ;4 3.4
Hz, H-30), 3.49 (m, 2H, H-20, OCH2CH2(CH2)5CH3),
1.61 (m, 2H, OCH2CH2(CH2)5CH3), 1.37–1.22
(m, 13H, H-60, OCH2CH2(CH2)5CH3), 0.85 (t, 3H,
OCH2CH2(CH2)5CH3); 13C NMR (D2O): d 103.2
(C-10), 95.3 (C-1), 73.4 (C-30), 71.7 (C-40), 71.4 (C-50),
70.9 (C-20), 69.7 (C-3), 69.6 (C-4), 69.1 (C-5), 69.0
(OCH2CH2(CH2)5CH3), 68.5 (C-6), 68.5 (C-2), 31.4
(OCH2CH2(CH2)5CH3), 28.9 (OCH2CH2(CH2)5CH3),
3.53 (dd, 1H, J2 ;3 10.0 Hz, H-20), 3.47 (dd, 1H, J2,3
9.7 Hz, H-2), 1.60 (m, 2H, OCH2CH2(CH2)5CH3),
1.35–1.23 (m, 13H, H-60, OCH2CH2(CH2)5CH3),
0.84 (t, 3H, OCH2CH2(CH2)5CH3); 13C NMR (D2O):
d 103.5 (C-10), 103.0 (C-1), 77.8 (C-4), 75.3 (C-5), 72.9
(C-30), 72.5 (C-3), 72.1 (C-2), 71.8 (C-40), 71.3
(C-20), 71.1 (C-50), 70.9 (OCH2CH2(CH2)5CH3), 60.7
(C-6), 31.3 (OCH2CH2(CH2)5CH3), 29.1 (OCH2-
CH2(CH2)5CH3), 28.6 (OCH2CH2(CH2)5CH3), 25.4
(OCH2CH2(CH2)5CH3), 22.3 (OCH2CH2(CH2)5CH3),
15.8 (C-60), 13.9 (OCH2CH2(CH2)5CH3); ESIMS
m/z calcd for [C20H38O10]Na+: 461.2357. Found:
461.2347.
0
0
28.6
(OCH2CH2(CH2)5CH3),
25.6
(OCH2CH2-
(CH2)5CH3), 22.1 (OCH2CH2(CH2)5CH3), 15.7 (C-60),
12.4 (OCH2CH2(CH2)5CH3); ESIMS m/z calcd for
[C20H38O10]Na+: 461.2357. Found: 461.2349.
3.17.3. Octyl a-L-fucopyranosyl-(1!6)-b-D-galacto-
pyranoside (29-iii). 1H NMR (D2O): d 4.93 (d, 1H,
0
0
3.17. Synthesis of octyl L-fucopyranosyl-(1!6)-D-galacto-
J1 ;2 3.9 Hz, H-10), 4.38 (d, 1H, J1,2 7.9 Hz, H-1), 4.09
0
0
pyranosides (29)
(q, 1H, J5 ;6 6.1 Hz, H-50), 3.91 (br d, 1H, J3,4 3.3 Hz,
H-4), 3.89–3.75 (m, 7H, H-20, H-3, H-4, H-5, H-6a,
H-6b, OCH2CH2(CH2)5CH3), 3.69–3.61 (m, 2H, H-3,
OCH2CH2(CH2)5CH3), 3.50 (dd, 1H, J2,3 9.7 Hz, H-2),
1.61 (m, 2H, OCH2CH2(CH2)5CH3), 1.35–1.26 (m,
The title compound was synthesized according to the
procedure described for the synthesis of compound 26.
Overall yield: 68% (three steps after Sep-Pak). Ratio:
29-i:29-ii:29-iii:29-iv 1:6.4:1.9:15 (estimated by LC–MS
based on assumption that these compounds are similarly
ionized). HPLC conditions follows; flow rate: 1.0 mL/
min; column: Xterra RP18, 5 lm, 250 mm (L) ·
4.6 mm (I.D.); mobile phase: CH3CN/H2O 20:80; col-
umn temperature: 30 ꢁC; detection (m/z): 461.3
[M+Na]+; retention times (min): 39.7 (29-i; a-Fuc-a-
Gal), 46.9 (29-iii; a-Fuc-b-Gal), 78.7 (29-ii; b-Fuc-a-
Gal), and 82.7 (29-iv; b-Fuc-b-Gal).
0
0
10H, OCH2CH2(CH2)5CH3), 1.22 (d, 3H, J5 ;6 6.6 Hz,
H-60), 0.85 (t, 3H, OCH2CH2(CH2)5CH3); 13C NMR
(D2O): d 103.2 (C-1), 100.0 (C-10), 74.0 (C-5), 73.2
(C-3), 72.1 (C-40), 71.2 (OCH2CH2(CH2)5CH3), 71.1
(C-2), 70.1 (C-30), 69.2 (C-4), 68.6 (C-20), 68.4
(C-6), 67.1 (C-50), 31.2 (OCH2CH2(CH2)5CH3), 29.0
(OCH2CH2(CH2)5CH3), 28.6 (OCH2CH2(CH2)5CH3),
25.1 (OCH2CH2(CH2)5CH3), 22.1 (OCH2CH2(CH2)5-
CH3), 15.8 (C-60), 12.4 (OCH2CH2(CH2)5CH3); ESIMS

I. Ohtsuka et al. / Carbohydrate Research 341 (2006) 1476–1487
1487
m/z calcd for [C20H38O10]Na+: 461.2357. Found:
461.2357.
10. Kanie, O.; Barresi, F.; Ding, Y.; Labbe, J.; Otter, A.;
Forsberg, L. S.; Ernst, B.; Hindsgaul, O. Angew. Chem.,
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3.17.4. Octyl b-L-fucopyranosyl-(1!6)-b-D-galactopyran-
oside (29-iv). 1H NMR (D2O): d 4.39 (m, 2H, J1,2
7.9 Hz, J1 ;2 7.9 Hz, H-1,10), 4.01 (dd, 1H, J5,6a 8.6 Hz,
J6a,6b 12.6 Hz, H-6a), 3.95 (br d, 1H, J3,4 3.4 Hz, H-4),
3.92–3.83 (m, 3H, H-5, H-6b, OCH2CH2(CH2)5CH3),
0
0
3.77 (q, 1H, J5 ;6 6.6 Hz, H-50), 3.73 (br d, 1H, J3 ;4
0
0
0
0
3.3 Hz, H-40), 3.67–3.62 (m, 3H, H-3, H-30,
OCH2CH2(CH2)5CH3), 3.48 (m, 2H, H-2,20), 1.60
(m, 2H, OCH2CH2(CH2)5CH3), 1.35–1.24 (m, 13H,
H-60,OCH2CH2(CH2)5CH3), 0.84 (t, 3H, OCH2CH2-
(CH2)5CH3); 13C NMR (D2O): d 103.3 (C-1), 103.3
(C-10), 73.5 (C-5), 73.1 (C-3), 73.1 (C-30), 71.7 (C-40),
71.4 (C-50), 71.1 (OCH2CH2(CH2)5CH3), 70.9 (C-2),
70.9 (C-20), 69.1 (C-4), 69.1 (C-6), 31.3 (OCH2-
CH2(CH2)5CH3), 28.9 (OCH2CH2(CH2)5CH3), 28.6
(OCH2CH2(CH2)5CH3), 25.1 (OCH2CH2(CH2)5CH3),
22.1 (OCH2CH2(CH2)5CH3), 15.7 (C-60), 13.6 (OCH2-
CH2(CH2)5CH3); ESIMS m/z calcd for [C20H38O10]-
Na+: 461.2357. Found: 461.2354.
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Acknowledgements
This work was supported in part by the Key Technology
Research Promotion Program, the New Energy and
Industrial Development Organization (NEDO), the
Ministry of Economy, Trade, and Industry (METI) of
Japan and the Mitsubishi Chemical Corp.
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