Fluorescence modification of Gb3 oligosaccharide and rapid synthesis of oligosaccharide moieties using fluorous protective group

Article abstract of DOI:10.1016/j.tet.2005.05.001

The use of the bisfluorous chain-type propanoyl (Bfp) group as a fluorous protective group made it possible to rapidly synthesize the Gb2 and Gb3 oligosaccharide derivatives by a simple fluorous-organic extraction purification. Furthermore, the fluorescence-labeled Gb2 and Gb3 oligosaccharides were prepared as a potential Vero Toxins detecting reagent.

Full text of DOI:10.1016/j.tet.2005.05.001

Tetrahedron 61 (2005) 6518–6526
Fluorescence modification of Gb3 oligosaccharide
and rapid synthesis of oligosaccharide moieties using
fluorous protective group
a
a
a
a
Tsuyoshi Miura,a,b, Satoshi Tsujino, Ai Satoh, Kohtaro Goto, Mamoru Mizuno,
*
Midori Noguchi,a Tetsuya Kajimoto,c Manabu Node,c Yasuoki Murakami,b Nobuyuki Imaib
and Toshiyuki Inazua,d,
*
aThe Noguchi Institute, 1-8-1 Kaga, Itabashi-ku, Tokyo 173-0003, Japan
bFaculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025, Japan
cKyoto Pharmaceutical University, 1 Shichono-cho, Misasagi, Yamashina-ku, Kyoto 607-8412, Japan
dDepartment of Applied Chemistry, School of Engineering, and Institute of Glycotechnology, Tokai University, Kitakaname 1117, Hiratsuka,
Kanagawa 259-1292, Japan
Received 24 March 2005; revised 30 April 2005; accepted 2 May 2005
Available online 23 May 2005
Abstract—The use of the bisfluorous chain-type propanoyl (Bfp) group as a fluorous protective group made it possible to rapidly synthesize
the Gb2 and Gb3 oligosaccharide derivatives by a simple fluorous-organic extraction purification. Furthermore, the fluorescence-labeled Gb2
and Gb3 oligosaccharides were prepared as a potential Vero Toxins detecting reagent.
q 2005 Elsevier Ltd. All rights reserved.
1. Introduction
were substituted with simple lipophilic fluorescence groups,
such as a dansyl or fluorescein group, for the ceramide
moieties of the glycolopids.
Vero toxins (VTs) are the Shiga-like toxins produced by
Escherichia coli O-157:H-7.1 VTs cause serious clinical
complications in humans. VTs initially produce diarrhea in
human beings, then further progresses to the Hemolytic
Uremic Syndrome, renal failure, and finally death. In order to
suppress the infection progress by E. coli O-157:H-7 to a
minimum, the development of an accurate and rapid detecting
method of VTs is essential. VTs are AB5 toxins with an
enzymatically active A-subunit and a cell binding B-subunit.
The B-subunit mainly recognizes a galactobiosyl a-(1/4)-
linkage in the glycolipid Gb2 [Gala-(1/4)Gal-Cer] and Gb3
[Gala-(1/4)Galb-(1/4)Glc-Cer], and VTs induce the
carbohydrate-mediated internalization into the host cell.2 A
carbohydrate–lectin interaction in the solution phase was
sensitively detected by fluorescence polarization using a
fluorescence-labeled carbohydrate.3 Recently, we reported
that fluorescence polarization detected the carbohydrate–
lectin interaction using the fluorescence-labeled oligosacchar-
ides derived from glycosyl amino acids.4 To develop a method
sensitively detecting VTs, we attempted the synthesis of
fluorescence-labeled Gb2 and Gb3 oligosaccharides, which
The synthesis of the Gb2 and Gb3 oligosaccharides have
already been accomplished by several groups.5 However,
each synthetic method requires much time and cost due to
the purification procedure, such as column chromatography
in multisteps. The heavy fluorous technique using a fluorous
protecting group developed by Curran et al. is an excellent
methodology to resolve these problems.6 A highly fluorin-
ated compound, in which a fluorous protecting group is
introduced, is readily separated from nonfluorinated com-
pounds by a simple fluorous-organic phase separation
without column chromatography. Several fluorous protect-
ing groups were developed for the fluorous synthesis.7 We
also have reported the fluorous oligosaccharide synthesis
using the Bfp (bisfluorous chain-type propanoyl) group as a
*
Corresponding authors. Tel.: C81 479304612; fax: C81 479304610
(T.M.); e-mail: tmiura@cis.ac.jp
0040–4020/$ - see front matter q 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2005.05.001

T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
6519
novel fluorous protective group,8 the rapid synthesis of the
Gb2 and Gb3 oligosaccharides using the Bfp group in a
preliminary communication,9 and the synthesis of oligo-
saccharidesandpeptidesusingthefluoroussupports.10 Herein,
we would like to describe the full details of the rapid synthesis
of the Gb2 and Gb3 oligosaccharides using the Bfp group as a
fluorous protective group and the fluorescence modification of
the Gb2 and Gb3 oligosaccharides.
the mixture between FC-72 and MeOH. The methyl ester of
Bfp (Bfp-OMe, 1b) was extracted from the FC-72 layer and
treated with aqueous NaOH to recover 1a, which is able to
be reused as a fluorous protective reagent. Finally, the pure
galabiose derivative 8 was obtained from the single silica
gel column chromatographic purification step in a 38%
overall yield from 2 (five steps). The disaccharide 8 was
converted to the acetate 9 by treatment with acetic
anhydride in pyridine for identification of the structure.
Next, we synthesized the Gb3 trisaccharide 15 as described
in Scheme 2. The Bfp group was introduced to the four
hydroxyl functions of the lactose derivative 10 using DCC
and DMAP to give the fluorous compound 11.11 The
benzylidene group and trityl (Tr) group of 11 were
deprotected by treatment with HCl in AcOEt–EtOC4F9 to
afford 12.11 The benzoyl (Bz) group was selectively
introduced to the two primary hydroxyl functions of 12 to
obtain the fluorous glycosyl acceptor 13.11 The reaction of
the fluorous glycosyl acceptor 13 with the glycosyl donor 6
(5 equiv) in the presence of TMSOTf in ether–EtOC4F9
selectively afforded only the a-linked fluorous trisaccharide
14.11 The fluorous intermediates 11, 12, 13, and 14 were
extracted with FC-72 by partitioning the product mixture
between FC-72 and an organic solvent (toluene or
methanol), and were purified enough without silica gel
column chromatography. The Bfp group of 14 was removed
by treatment with sodium methoxide in MeOH–ether to
afford 15, which was extracted with MeOH by partitioning
the crude residue between FC-72 and MeOH. Finally, the
pure trisaccharide 15 was obtained by only one silica gel
column chromatography purification during the final step in
a 43% overall yield from 10 (five steps). The trisaccharide
15 was converted to 16 by treatment with acetic anhydride
in pyridine for identification of the structure.
2. Results and discussion
We first synthesized the galabiose derivative 8 as shown in
Scheme 1. The Bfp-OH (1a)8 was attached to the hydroxyl
functions of the galactose derivative 2 using N,N0-
dicyclohexylcarbodiimide (DCC) and 4-(N,N-dimethylamino)
pyridine (DMAP) to give the fluorous compound 3.11 The
benzylidene group of 3 was removed by treatment with
camphorsulfonic acid (CSA) in MeOH–CHCl3 to afford the
corresponding product 4.11 The Bfp group was selectively
introduced into the primary hydroxyl function of 4 using
DCC and DMAP at K20 8C to give the fluorous glycosyl
acceptor 5.11 The fluorous disaccharide 711 was obtained by
the reaction of 5 with the glycosyl donor 6 (6.1 equiv) in the
presence of trimethylsilyl trifluoromethanesulfonate
(TMSOTf) in Et2O–EtOC4F9.12 The a-selectivity of the
glycosylation reaction was very high and no b-isomer could
be detected. The fluorous intermediates 3, 4, 5, and 7 were
each extracted with the fluorous solvent FC-7213 by
partitioning the product mixture between FC-72 and an
organic solvent, such as toluene or methanol. No further
purification, such as silica-gel column chromatography, was
carried out. The Bfp group of 7 was easily removed by
treatment with sodium methoxide in MeOH–Et2O to afford
the crude 8, which was extracted with MeOH by partitioning
Scheme 1. Synthesis of Gb2 oligosaccharide using fluorous protecting gorup Bfp.

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T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
Scheme 2. Synthesis of Gb3 oligosaccharide using fluorous protecting group Bfp.
Scheme 3. Intorduction of fluorescein groups to Gb2 oligosaccharide.

T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
6521
Scheme 4. Introduction of fluorescein groups to Gb3 oligosaccharide.
We then selected the dansyl and fluorescein groups as the
fluorescence group, and introduced them to the Gb2
oligosaccharide as indicated in Scheme 3. The disaccharide
8 was oxidized by ozonolysis, followed by condensation with
hydroxylamine to afford the corresponding imine 18 as an
inseparable mixture of syn and anti isomers in 77% yield (two
steps). The imine 18 was hydrogenated over Pd/C to give the
mixture of the amine 19 and the protected ones, which was
coupled with 20 to afford the desired Gb2 oligosaccharide 21
which possessed the dansyl group as the fluorescence group in
42% yield (two steps). The amine 19 was also reacted with 22
to give the Gb2 oligosaccharide 23 with a fluorescein group in
a 34% yield (two steps) (Scheme 4).
purification. The fluorous oligosaccharide synthesis can be
carried out on a large scale due to the liquid phase synthesis.
As each synthetic intermediate containing the Bfp group
was monitored by TLC, NMR, and MS, the reaction
conditions for each synthetic step could be rapidly
optimized. The fluorous intermediates could also be purified
by silica gel column chromatography when required
purification. After optimization of the reaction conditions
in each step, the synthesis in multisteps was accomplished
by a fluorous-organic partition purification without column
chromatography. The only final product, from which the
Bfp groups were removed, was purified by column
chromatography on silica gel. Therefore, the fluorous
oligosaccharide synthesis using the Bfp group is an
excellent strategic alternative to solid phase oligosaccharide
synthesis, and removes some of the disadvantages of the
solid phase method. The detection assay of VTs using the
fluorescence-labeled Gb2 and Gb3 oligosaccharides is now
in progress.
Finally, we synthesized the Gb3 oligosaccharide 27 and 28
by the similar procedure. The trisaccharide 15 was oxidized
by ozone to give the aldehyde 24, which was coupled with
hydroxylamine to afford the corresponding compound 25 as
an inseparable mixture of syn and anti isomers in 67% yield
(two steps). Compound 25 was reduced by catalytic
hydrogenation over Pd/C to give the mixture of the amine
26 and the protected ones, which was coupled with 20 to
afford the desired Gb3 oligosaccharide 27 with a dansyl
group in 25% yield (two steps). The amine 26 was also
reacted with 22 to give the Gb3 oligosaccharide 28 with a
fluorescein group in 35% yield (two steps).
4. Experimental
4.1. General
The 1H NMR spectra were recorded using JEOL JNM-EX-
400 (400 MHz) and JEOL JNM-ECA-600 (600 MHz)
spectrometers. The Mass spectra (MS) of the compounds
with a high molecular weight were recorded using a
MALDI-TOF-MS (Voyager-DE STR) spectrometer. The
high-resolution MS (HRMS) were recorded on an ESI-TOF-
MS (Marinere) spectrometer. Part of the products was
isolated by column chromatography on silica gel (Kanto
Chemical, silica gel 60N, spherical, neutral, 40–50 mm).
The fluorous solvent FC-72 and Novec HFE-7200 were
3. Conclusion
In conclusion, we succeeded in synthesizing the Gb2 and
Gb3 oligosaccharides containing fluorescence groups. The
use of the Bfp group as a fluorous protecting group made
it possible to rapidly synthesize the Gb2 and Gb3 oligo-
saccharide derivatives by a fluorous-organic extraction

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T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
purchased from 3 M Tokyo. Bfp-OH (1a)8 is commercially
available from Kokusan Chemical.
(c 3.02, FC-72); 1H NMR (400 MHz, CDCl3): dZ1.87 (6H,
m), 2.10 (6H, m), 2.57 (19H, m), 3.43–3.98 (13H, m), 4.10
(2H, m), 4.34 (3H, m), 4.49 (1H, m), 4.75 (1H, m), 5.26 (3H,
m), 5.82 (1H, m); MALDI-TOF MS found: m/z [MCNa]C
3257.5. Calcd for C84H55F102N3O10Na [MCNa]C 3258.2.
Found: m/z [MCK]C 3274.2. Calcd for C84H55F102N3O10K
[MCK]C 3274.2.
4.1.1. Compound 3. DMAP (3.27 g, 26.8 mmol) and DCC
(9.91 g, 48.1 mmol) were added to a solution of 214 (2.50 g,
8.12 mmol) and 1a8 (19.0 g, 18.6 mmol) in dried CH2Cl2
(200 mL). After stirring for 5 h at rt, MeOH (100 mL) was
added to the reaction mixture. The mixture was stirred for
30 min at rt, and the CH2Cl2 was evaporated. The reaction
mixture was extracted three times with FC-72. The FC-72
layers were combined and evaporated to give a crude 3
(21.1 g). The crude 3 was used in the next step without
further purification. The authentic sample was obtained as a
colorless amorphous solid by purifying part of the sample
(silica gel column chromatography with a 2:1 mixture of
hexane and AcOEt). RfZ0.19 (hexane–AcOEtZ2:1); [a]D26
4.1.4. Compound 7. Molecular sieves 4A powder (3.3 g)
was added to a solution of the crude 5 (1.26 g) and 6 (1.63 g,
2.38 mmol) in anhydrous ether (40 mL)–EtOC4F9 (20 mL)
under an argon atmosphere. After stirring for 3 h at rt,
TMSOTf (210 mL, 1.16 mmol) was added at 0 8C to the
reaction mixture. The mixture was stirred for 30 min at 0 8C,
and filtered on Celite. The filtrate was added to saturated
aqueous NaHCO3 and extracted three times with AcOEt.
The AcOEt layers were combined, washed with brine, dried
over anhydrous Na2SO4, and evaporated. The residue was
dissolved in MeOH and FC-72, and extracted three times
with FC-72. The FC-72 layers were combined and
evaporated to afford a crude 7 (1.28 g). The crude 7 was
used in the next step without further purification. The
authentic sample was obtained as a colorless amorphous
solid by purifying part of the crude 7 (silica gel column
chromatography, eluent: hexane–AcOEtZ2:1). RfZ0.37
1
C12.8 (c 1.27, CHCl3); H NMR (400 MHz, CDCl3): dZ
1.74–2.08 (8H, m), 2.44–2.68 (12H, m), 3.14–3.69 (9H, m),
4.09 (2H, m), 4.37 (3H, m), 4.58 (1H, m), 4.95 (1H, m), 5.19
(1H, d, JZ10.8 Hz), 5.27 (1H, d, JZ17.2 Hz), 5.41 (1H, t,
JZ7.6 Hz), 5.50 (1H, s), 5.85 (1H, m), 7.37 (3H, m), 7.51
(2H, m); MALDI-TOF MS found: m/z [MCNa]C 2342.5.
Calcd for C66H46F68N2O10Na [MCNa]C 2341.2. Found:
m/z [MCK]C 2359.3. Calcd for C66H46F68N2O10K [MC
K]C 2357.2.
1
(hexane–AcOEtZ2:1); [a]2D6 C11.5 (c 2.33, FC-72); H
4.1.2. Compound 4. CSA (1.63 g, 7.0 mmol) was added to a
solution of the crude 3 (10.7 g) in CHCl3 (80 mL)–MeOH
(40 mL). After stirring for 5 h at rt, toluene (100 mL) and
saturated aqueous NaHCO3 (100 mL) were added to the
reaction mixture. The mixture was stirred for 30 min at rt,
and the organic solvents (CHCl3 and MeOH) were
evaporated. The reaction mixture was then extracted three
times with FC-72. The FC-72 layers were combined, dried
over anhydrous Na2SO4, and evaporated to afford a crude 4
(9.54 g). The crude 4 was used in the next step without
further purification. The authentic sample was obtained as a
colorless amorphous solid by purifying part of the crude 4
(silica gel column chromatography, eluent; hexane–
AcOEtZ1:1). RfZ0.30 (hexane–AcOEtZ1:1); [a]2D6 K1.1
(c 1.25, FC-72); 1H NMR (400 MHz, CDCl3): dZ1.87 (4H,
m), 2.09 (4H, m), 2.58 (14H, m), 3.38–3.68 (8H, m), 3.78–
4.18 (4H, m), 4.33 (2H, m), 4.52 (1H, d, JZ8.1 Hz), 4.78
(1H, m), 5.17 (1H, d, JZ10.3 Hz), 5.25 (1H, d, JZ
17.3 Hz), 5.34 (1H, m), 5.85 (1H, m); MALDI-TOF MS
found: m/z [MCNa]C 2251.6. Calcd for C59H42F68N2O10-
Na [MCNa]C 2253.2. Found: m/z [MCK]C 2267.8. Calcd
for C59H42F68N2O10K [MCK]C 2269.1.
NMR (400 MHz, CDCl3): dZ1.87 (6H, m), 2.04 (6H, m),
2.54 (18H, m), 3.21–3.71 (15H, m), 4.06 (5H, m), 4.27–4.55
(8H, m), 4.67–4.94 (7H, m), 5.18 (1H, d, JZ10.5 Hz), 5.27
(2H, m), 5.84 (1H, m), 7.27 (20H, m); MALDI-TOF MS
found: m/z [MCNa]C 3781.5. Calcd for C118H89F102N3-
O17Na [MCNa]C 3780.5. Found: m/z [MCK]C 3797.4.
Calcd for C118H89F102N3O17K [MCK]C 3796.4.
4.1.5. Compound 8. A 28% solution of sodium methoxide
in MeOH (140 mL) was added to a solution of the crude 7
(1.28 g) in ether (15 mL)–MeOH (15 mL). After stirring for
3 h at rt, the reaction mixture was neutralized with
Amberlite IR-120 (HC form), and filtered. The filtrate was
evaporated. The residue was dissolved in MeOH and FC-72,
and extracted three times with FC-72. The methanol layer
was evaporated to give a crude 8. The FC-72 layers were
combined and evaporated to afford the pure compound 1b.8b
The crude 8 was purified by column chromatography on
silica gel with a 1:2 mixture of hexane and AcOEt to give
the pure 8 (114 mg, 38% in five steps from 2) as colorless
1
crystals. Mp 68–70 8C; [a]2D6 C33.1 (c 1.45, MeOH); H
NMR (400 MHz, CDCl3): dZ2.10 (2H, brs), 3.27 (1H, dd,
JZ9.3, 3.4 Hz), 3.37 (1H, m), 3.45 (1H, dd, JZ10.0,
7.6 Hz), 3.56 (1H, t, JZ9.0 Hz), 3.70 (4H, m), 3.90 (1H,
brs), 3.94 (1H, brs), 4.01 (1H, dd, JZ10.0, 2.4 Hz), 4.13
(3H, m), 4.24 (1H, d, JZ7.3 Hz), 4.35 (1H, m), 4.37 (1H, d,
JZ11.2 Hz), 4.45 (1H, d, JZ11.2 Hz), 4.53 (1H, d, JZ
11.5 Hz), 4.72 (1H, d, JZ12.7 Hz), 4.77 (2H, s), 4.82 (1H,
d, JZ3.4 Hz), 4.91 (2H, d, JZ11.5 Hz), 5.21 (1H, d, JZ
10.3 Hz), 5.32 (1H, d, JZ17.3 Hz), 5.94 (1H, m), 7.30
(20H, m); 13C NMR (100 MHz, CDCl3): dZ60.35, 69.58,
70.40, 71.36, 72.10, 72.68, 73.47, 73.72, 74.09, 74.35,
74.61, 74.95, 75.69, 79.07, 80.70, 100.85, 102.03, 117.83,
127.22, 127.65, 127.70, 127.82, 127.98, 128.03, 128.19,
128.32, 128.39, 128.43, 128.58, 133.66, 136.86, 137.31,
137.84, 137.86; HRMS (ESI-TOF): calcd for C43H50O11Na
4.1.3. Compound 5. DMAP (243 mg, 2.00 mmol) and DCC
(264 mg, 1.28 mmol) were added to a solution of the crude 4
(0.89 g) and 1a (436 mg, 0.43 mmol) in CH2Cl2 (20 mL)–
EtOC4F9 (20 mL) at K20 8C. After stirring for 27 h at
K20 8C, MeOH (6 mL) and toluene (50 mL) were added to
the reaction mixture. The mixture was stirred for 30 min at
rt, and then the CH2Cl2 was evaporated. The mixture was
extracted three times with FC-72. The FC-72 layers were
combined and evaporated to give a crude 5 (1.26 g). The
crude 5 was used in the next step without further
purification. The authentic sample was obtained as a
colorless amorphous solid by purifying part of the crude 5
(silica gel column chromatography, eluent: hexane–
AcOEtZ3:2). RfZ0.32 (hexane–AcOEtZ3:2); [a]2D6 K7.1

T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
6523
(MCNa)C: 765.3245. Found: 765.3220. Anal. Calcd for
C43H50O11H2O: C, 67.88; H, 6.89. Found: C, 67.55; H, 6.85.
(30 mL) was added to the reaction mixture. The mixture was
stirred for 1 h at rt, toluene was added to the reaction
mixture. The mixture was extracted three times with FC-72.
The FC-72 layers were combined and evaporated to give a
crude 11 (17.4 g). The crude 11 was used in the next step
without further purification. The authentic sample was
obtained by purifying part of the sample (silica gel column
chromatography, eluent: hexane–AcOEtZ2:1) as a color-
less amorphous solid. RfZ0.44 (hexane–AcOEtZ2:1);
4.1.6. Compound 9. Acetic anhydride (0.5 mL) was added
to a solution of 8 (13.7 mg, 18.5 mmol) in pyridine (1.0 mL).
After stirring for 15 h at rt, MeOH (10 mL) was added at
0 8C to the reaction mixture and evaporated. The residue
was purified by column chromatography on silica gel with a
7:4 mixture of hexane and AcOEt to give the pure 9
(15.2 mg, 95%) as a colorless oil. [a]2D2 C32.9 (c 0.83,
1
[a]2D3 K1.5 (c 1.05, AcOEt); H NMR (400 MHz, CDCl3):
1
dZ1.70–2.08 (16H, m), 2.34–2.72 (24H, m), 3.00–3.75
(20H, m), 4.00 (1H, m), 4.13–4.72 (8H, m), 4.96 (1H, m),
5.12 (2H, m), 5.21 (1H, d, JZ10.5 Hz), 5.29 (1H, d, JZ
17.3 Hz), 5.44 (1H, m), 5.92 (1H, m), 7.27 (3H, m), 7.33
(9H, m), 7.42 (2H, m), 7.50 (6H, m); MALDI-TOF MS
found: m/z [MCNa]C 4753.6. Calcd for C141H96F136N4-
O19Na [MCNa]C 4755.4.
CHCl3); H NMR (400 MHz, CD3OD): dZ1.90 (3H, s),
2.00 (3H, s), 2.03 (3H, s), 3.47 (1H, dd, JZ8.8, 5.1 Hz),
3.61 (1H, t, JZ8.8 Hz), 3.86 (1H, t, JZ6.3 Hz), 3.96 (1H,
dd, JZ10.0, 3.2 Hz), 4.08 (1H, d, JZ2.9 Hz), 4.11 (3H, m),
4.30 (2H, m), 4.35 (1H, dd, JZ11.5, 5.9 Hz), 4.43 (1H, dd,
JZ11.5, 7.1 Hz), 4.45 (1H, d, JZ12.0 Hz), 4.49 (1H, d, JZ
12.0 Hz), 4.51 (1H, d, JZ11.2 Hz), 4.61 (1H, d, JZ7.8 Hz),
4.70, (1H, d, JZ11.7 Hz), 4.76 (2H, s), 4.78 (1H, d, JZ
11.7 Hz), 4.79 (1H, d, JZ3.2 Hz), 4.85 (1H, d, JZ11.2 Hz),
4.96 (1H, dd, JZ10.5, 2.9 Hz), 5.16 (1H, dd, JZ10.5,
1.5 Hz), 5.23 (1H, dd, JZ10.5, 7.8 Hz), 5.27 (1H, dd, JZ
17.3, 1.5 Hz), 5.89 (1H, m), 7.31 (20H, m); 13C NMR
(100 MHz, CD3OD): dZ20.79, 20.81, 20.94, 63.92, 69.04,
70.73, 70.85, 70.99, 73.59, 73.91, 73.95, 74.31, 74.59,
76.08, 76.35, 76.82, 77.27, 79.76, 101.35, 101.75, 117.27,
128.45, 128.50, 128.64, 128.84, 129.08, 129.10, 129.19,
129.28, 129.29, 135.12, 139.40, 139.74, 140.00, 140.06,
171.16, 171.88, 172.11; MALDI-TOF MS found: m/z [MC
Na]C 889.9. Calcd for C49H56O14Na [MCNa]C 891.4.
Found: m/z [MCK]C 905.8. Calcd for C49H56O14K [MC
K]C 907.3.
4.1.9. Compound 12. To a solution of the crude 11 (6.71 g)
in EtOC4F9 (60 mL) were added 20 mL of a 4 M solution of
hydrochloric acid in AcOEt and 1 mL of water were added
at 0 8C. After stirring for 1 h at 0 8C, toluene and water were
added to the reaction mixture. The reaction mixture was
extracted three times with FC-72. The FC-72 layers were
combined, dried over anhydrous Na2SO4, and evaporated to
give a crude 12 (5.65 g). The crude 12 was used in the
next step without further purification. The authentic
sample was obtained by purifying part of the sample (silica
gel column chromatography, eluent: CHCl3–MeOHZ30:1)
as a colorless amorphous solid. Compound 12 is insoluble
in all deuterium solvents, therefore, no NMR spectra
could be obtained. RfZ0.26 (CHCl3–MeOHZ20:1); [a]D23
C0.8 (c 0.93, FC-72); MALDI-TOF MS found: m/z
[MCNa]C 4423.2. Calcd for C115H78F136N4O19Na [MC
Na]C 4425.3. Found: m/z [MCK]C 4440.3. Calcd for
C115H78F136N4O19K [MCK]C 4441.3.
4.1.7. Compound 10. Triphenylmethyl chloride (7.39 g,
26.5 mmol) was added at rt to a solution of allyl 40,60-O-
benzylidene-b-lactoside15 (4.13 g, 8.80 mmol) in pyridine
(50 mL). After stirring for 24 h at 50 8C, MeOH (2 mL) was
added to the reaction mixture. After cooling, the reaction
mixture was evaporated. The residue was treated with water
and extracted three times with AcOEt. The AcOEt layers
were combined, washed with brine, dried over anhydrous
Na2SO4, and evaporated. The crude product was purified by
column chromatography on silica gel with a 20:1 mixture of
CHCl3 and MeOH to afforded the pure 10 (3.88 g, 62%) as a
4.1.10. Compound 13. Benzoyl chloride (0.75 mL,
6.46 mmol) was added at K20 8C to a solution of the
crude 12 (6.65 g) and triethylamine (5.4 mL, 38.5 mmol) in
anhydrous CH2Cl2 (100 mL)–EtOC4F9 (100 mL). After
stirring for 15 h at K20 8C, MeOH (50 mL) was added to
the reaction mixture, and then the CH2Cl2 and EtOC4F9
were evaporated. MeOH was added to the mixture, and
the resulting solution was extracted three times with FC-72.
The FC-72 layers were combined and evaporated to give a
crude 13 (6.01 g). The crude 13 was used in the next step
without further purification. The authentic sample was
obtained as a colorless amorphous solid by purifying part
of the sample (silica gel column chromatography, eluent:
hexane–AcOEtZ1.8:1). RfZ0.71 (CHCl3–MeOHZ20:1);
colorless powder. [a]2D3 K25.6 (c 1.04, CHCl3); H NMR
1
(400 MHz, CDCl3): dZ1.68 (1H, d, JZ3.4 Hz), 2.38 (1H,
d, JZ8.8 Hz), 2.56 (1H, d, JZ2.0 Hz), 3.30 (1H, dd, JZ
10.5, 4.4 Hz), 3.38 (1H, dd, JZ9.5, 3.7 Hz), 3.42 (1H, s),
3.52 (2H, m), 3.63 (3H, m), 3.74 (1H, t, JZ9.0 Hz), 4.04
(1H, d, JZ12.9 Hz), 4.13 (1H, d, JZ3.2 Hz), 4.17 (1H, d,
JZ7.6 Hz), 4.26 (3H, m), 4.43 (1H, d, JZ7.6 Hz), 4.46
(1H, dd, JZ12.9, 5.4 Hz), 5.27 (1H, d, JZ10.5 Hz), 5.38
(1H, d, JZ17.1 Hz), 5.50 (1H, s), 6.04 (1H, m), 7.28 (9H,
m), 7.36 (3H, m), 7.47 (8H, m); 13C NMR (100 MHz,
CDCl3): dZ62.40, 66.64, 68.59, 69.96, 71.02, 72.29, 73.52,
73.83, 74.55, 74.65, 79.70, 86.38, 101.03, 102.60, 117.82,
125.99, 126.77, 127.49, 127.95, 128.45, 128.82, 133.53,
136.90, 143.43; HRMS (ESI-TOF): calcd for C41H44O11Na
(MCNa)C: 735.2776. Found: 735.2814.
1
[a]2D3 3.3 (c 1.13, AcOEt); H NMR (400 MHz, CDCl3):
dZ1.85 (8H, m), 2.06 (8H, m), 2.57 (25H, m), 3.38–
3.75 (17H, m), 3.88 (2H, m), 4.07 (2H, m), 4.27 (2H,
m), 4.41–4.98 (7H, m), 5.16 (4H, m), 5.80 (1H, m),
7.41–7.60 (6H, m), 8.00 (4H, m); MALDI-TOF MS
found: m/z [MCNa]C 4632.5. Calcd for C129H86F136-
N4O21Na [MCNa]C 4633.4. Found: m/z [MCK]C
4648.4. Calcd for C129H86F136N4O21K [MCK]C 4649.3.
4.1.8. Compound 11. DMAP (3.28 g, 26.8 mmol) and DCC
(7.90 g, 38.3 mmol) were added to a solution of 10 (2.73 g,
3.83 mmol) and 1a (16.5 g, 16.1 mmol) in anhydrous
CH2Cl2 (150 mL). After stirring for 3 h at rt, MeOH
4.1.11. Compound 14. Molecular sieves 4A powder (4.0 g)
was added to a solution of the crude 13 (1.85 g) and 6
(1.92 g, 2.81 mmol) in anhydrous ether (40 mL)–EtOC4F9

6524
T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
(40 mL) under an argon atmosphere. After stirring for
further 3 h at rt, TMSOTf (145 mL, 0.81 mmol) was added at
0 8C to the reaction mixture. The mixture was stirred for
30 min at 0 8C, quenched with triethylamine (1 mL), and
filtered on Celitection mixture. The filtrate was treated
with saturated aqueous NaHCO3, and extracted three times
with AcOEt. The AcOEt layers were combined, washed
with brine, dried over anhydrous Na2SO4, and evaporated.
The residue was dissolved in MeOH and FC-72, and
extracted three times with FC-72. The FC-72 layers were
combined and evaporated to give a crude 14 (1.74 g). The
crude 14 was used in the next step without further
purification. The authentic sample was obtained by purify-
ing part of the sample (silica gel column chromatography,
eluent: hexane–AcOEtZ5:2) as a colorless amorphous
solid. RfZ0.60 (hexane–AcOEtZ2:1); [a]2D3 C10.1 (c
gel with a 1:1 mixture of hexane and AcOEt to give the pure
product 16 (45.3 mg, 95%) as a colorless oil. [a]2D3 33.5
(c 2.11, CHCl3); 1H NMR (400 MHz, CDCl3): dZ1.85 (3H,
s), 1.87 (3H, s), 2.04 (9H, s), 2.12 (3H, s), 3.42 (1H, dd, JZ
8.8, 4.9 Hz), 3.60 (3H, m), 3.74 (1H, t, JZ9.5 Hz), 3.97
(1H, d, JZ2.9 Hz), 4.05 (4H, m), 4.15 (1H, brs), 4.28 (2H,
m), 4.43 (6H, m), 4.50 (1H, d, JZ7.8 Hz), 4.55 (1H, d, JZ
11.0 Hz), 4.66 (1H, d, JZ11.5 Hz), 4.75 (2H, d, JZ
11.2 Hz), 4.76 (1H, d, JZ4.2 Hz), 4.83 (2H, brs), 4.92 (2H,
m), 5.14 (2H, m), 5.19 (1H, d, JZ11.2 Hz), 5.25 (1H, d, JZ
17.3 Hz), 5.83 (1H, m), 7.24–7.43 (20H, m); 13C NMR
(100 MHz, CDCl3): dZ20.60, 20.73, 20.84, 20.88, 61.02,
61.90, 67.51, 69.49, 69.56, 69.88, 71.34, 72.24, 72.36,
72.47, 72.61, 72.73, 73.26, 74.15, 74.52, 74.88, 75.28,
75.87, 76.02, 79.01, 99.32, 100.92, 101.23, 117.44, 127.22,
127.29, 127.51, 127.56, 127.88, 127.97, 128.01, 128.11,
128.15, 128.18, 128.29, 133.19, 137.75, 138.96, 138.58,
138.64, 168.60, 169.40, 169.83, 170.19, 170.24, 170.52;
MALDI-TOF MS found: m/z [MCNa]C 1179.3. Calcd for
C61H72O22Na [MCNa]C 1179.4. Found: m/z [MCK]C
1195.8. Calcd for C61H72O22K [MCK]C1195.4.
1
1.09, AcOEt); H NMR (400 MHz, CDCl3): dZ1.68–2.07
(m, 16H), 2.35–2.69 (m, 24H), 3.20–3.90 (m, 21H), 4.08 (m,
5H), 4.28 (m, 4H), 4.40 (m, 1H), 4.52 (m, 4H), 4.70–4.98
(m, 10H), 5.17 (m, 4H), 5.80 (m, 1H), 7.22 (m, 18H), 7.40
(m, 4H), 7.50 (m, 2H), 7.59 (m, 2H), 7.99 (m, 4H); MALDI-
TOF MS found: m/z [MCNa]C 5157.5. Calcd for C163
-
H120F136N4O26Na [MCNa]C 5155.6. Found: m/z [MCK]C
5173.2. Calcd for C163H120F136N4O26K [MCK]C 5171.6.
4.1.14. Compound 17. A solution of 8 (58.6 mg, 79 mmol)
was treated with ozone at K78 8C by bubbling until the
solution remained blue. The reaction mixture was evacuated
with air for 5 min to remove the ozone, and then dimethyl
sulfide (0.2 mL) was added at K78 8C to the reaction
mixture. The resulting solution was allowed to warm to rt,
and then evaporated to afford a crude 17 (60.4 mg). The
crude 17 was used in the next step without further
purification due to its lability.
4.1.12. Compound 15. To a solution of the crude 14 (1.72 g,
0.36 mmol) in ether (30 mL)–MeOH (20 mL) was added
200 mL of a 28% solution of sodium methoxide in methanol.
After stirring for 2 h at rt, the reaction mixture was
neutralized with Amberlite IR-120 (HC form). After
filtration, the filtrate was evaporated. The residue was
dissolved in MeOH and FC-72, and extracted three times
with FC-72. The FC-72 layers were combined and
evaporated to afford the pure product 1b. The methanol
layer was evaporated to give a crude 15. The crude 15 was
purified by column chromatography on silica gel with a 15:1
mixture of CHCl3 and MeOH to give the pure product 15
(174 mg, 43% in five steps from 10) as a colorless
amorphous solid. Mp 128–130 8C; [a]2D3 C25.0 (c 1.53,
CHCl3: MeOHZ1:1); 1H NMR (400 MHz, CDCl3:
CD3ODZ1:1): dZ3.37–3.62 (8H, m), 3.69 (2H, m), 3.81
(1H, m), 3.87 (2H, m), 3.98 (2H, m), 4.07 (1H, dd, JZ10.3,
3.7 Hz), 4.17 (2H, m), 4.36 (1H, d, JZ7.1 Hz), 4.39 (1H,
m), 4.43 (1H, d, JZ11.7 Hz), 4.51 (1H, d, JZ11.7 Hz), 4.54
(1H, d, JZ11.7 Hz), 4.72 (1H, d, JZ11.7 Hz), 4.78 (2H, s),
4.85 (1H, d, JZ11.7 Hz), 4.89 (1H, d, JZ11.7 Hz), 4.91
(1H, d, JZ3.7 Hz), 5.20 (1H, dd, JZ10.3, 1.5 Hz), 5.34
(1H, dd, JZ17.1, 1.5 Hz), 5.96 (1H, m), 7.33 (20H, m); 13C
NMR (100 MHz, CDCl3: CD3ODZ1:1): dZ61.34, 61.70,
69.46, 70.69, 71.33, 72.01, 73.14, 73.72, 73.92, 74.11,
74.73, 74.89, 75.12, 75.21, 75.37, 76.49, 79.04, 80.69,
81.34, 100.89, 102.18, 104.57, 117.68, 127.81, 127.96,
128.06, 128.14, 128.33, 128.53, 128.55, 128.57, 128.69,
128.71, 128.76, 134.21, 137.82, 137.88, 138.35, 138.50;
HRMS (ESI-TOF): calcd for C49H60O16Na (MCNa)C:
927.3774. Found: 927.3750. Anal. Calcd for C49H60O161/
2H2O: C, 64.39; H, 6.73. Found: C, 64.45; H, 6.58.
4.1.15. Compound 18. Hydroxylamine hydrochloric acid
salt (1.5 mg, 21.7 mmol) was added at rt to a solution of the
crude 17 (10.5 mg, 14.1 mmol) in MeOH (15 mL)–H2O
(2 mL). After stirring for 1.5 h at rt, the MeOH was
evaporated. The resulting mixture was treated with water
and extracted three times with AcOEt. The AcOEt layers
were combined, washed with brine, dried over anhydrous
Na2SO4, and evaporated. The residue was purified by
column chromatography on silica gel with a 20:1 mixture of
CHCl3 and MeOH to give an inseparable mixture of E- and
Z-isomers 18 (8.2 mg, 77% in two steps) as a colorless
amorphous solid.
4.1.16. Compound 21. A solution of the mixture 18
(8.2 mg, 10.8 mmol) in EtOH (4 mL)–AcOH (1 mL) and
hydrochloric acid in 1,4-dioxane (4 M, 5 mL) were added at
rt to a suspension of 10% Pd/C (15 mg) in EtOH (3 mL).
After stirring for 22 h under a hydrogen atmosphere
(10 atm), the reaction mixture was filtered. The filtrate
was evaporated to give the crude compound 19 (8.7 mg). To
a solution of the crude 19 (8.7 mg) in 1,4-dioxane (2 mL)–
H2O (2 mL) were added 20 (20.3 mg, 43.9 mmol) and
triethylamine (6.4 mL, 46 mmol) at rt. After stirring for 3.5 h
at rt, the reaction mixture was dissolved in AcOEt and
water, and extracted three times with water. The water
layers were combined and evaporated to half to original
volume. The water layer was poured into Diaion HP-20. The
resin was washed with water, and then eluted with MeOH.
The MeOH fractions were then collected and evaporated.
The residue was purified by column chromatography on
silica gel with a 7:3:0.4 mixture of CHCl3, MeOH, and H2O
4.1.13. Compound 16. Acetic anhydride (1.5 mL) was
added to a solution of 15 (37.3 mg, 41.2 mmol) in pyridine
(3.0 mL). After stirring for 17 h at rt, MeOH (5 mL) was
added at 0 8C to the reaction mixture and evaporated. The
residue was purified by column chromatography on silica

T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
6525
to give the pure 21 (3.3 mg, 42% in two steps) as a green oil.
[a]2D5 C47.1 (c 0.51, MeOH); 1H NMR (400 MHz,
CD3OD): dZ1.16 (m, 2H), 1.34 (m, 4H), 2.01 (t, JZ
7.6 Hz, 2H), 2.81 (t, JZ6.8 Hz, 2H), 2.86 (s, 6H), 3.42 (m,
1H), 3.48 (dd, JZ10.3, 7.3 Hz, 1H), 3.54 (dd, JZ10.0,
2.9 Hz, 1H), 3.61 (m, 3H), 3.67 (m, 2H), 3.75 (m, 4H), 3.89
(m, 2H), 3.97 (d, JZ2.9 Hz, 1H), 4.25 (m, 1H), 4.27 (d, JZ
7.3 Hz, 1H), 4.94 (d, JZ3.4 Hz, 1H), 7.25 (d, JZ7.6 Hz,
1H), 7.56 (m, 2H), 8.16 (d, JZ7.4 Hz, 1H), 8.33 (d, JZ
8.8 Hz, 1H), 8.54 (d, JZ8.5 Hz, 1H); 13C NMR (100 MHz,
CD3OD): dZ26.32, 27.13, 30.32, 36.81, 40.47, 43.70,
45.83, 61.09, 62.62, 69.84, 70.68, 71.02, 71.32, 72.70,
72.82, 74.59, 76.16, 79.31, 102.57, 105.11, 116.33, 120.52,
124.24, 128.96, 130.05, 130.88, 130.98, 131.10, 137.06,
153.05, 175.95; HRMS (ESI-TOF): calcd for
C32H50N3O14S (MCH)C: 732.3008. Found: 732.2991.
4.1.20. Compound 26. A solution of the mixture 25
(14.8 mg, 16 mmol) in EtOH (5 mL)–AcOH (1 mL) and
hydrochloric acid in 1,4-dioxane (4 M, 5 mL) were added at
rt to a suspension of 10% Pd/C (18 mg) in EtOH (5 mL).
After stirring for 3 days under a hydrogen atmosphere
(10 atm), the reaction mixture was filtered. The filtrate was
evaporated to give a crude 26 (20.5 mg). The crude 26 was
used in the next step without further purification.
4.1.21. Compound 27. To a solution of the crude 26
(20.5 mg) in 1,4-dioxane (3 mL)–H2O (3 mL) were added
20 (19.6 mg, 42 mmol) and triethylamine (5 mL, 36 mmol) at
rt. After stirring for 22 h at rt, the reaction mixture was
dissolved in AcOEt and water, and extracted three times
with water. The water layers were combined and evaporated
to half to original volume. The water layer was poured into
Diaion HP-20. The resin was washed with water, and then
eluted with MeOH. The MeOH fractions were then
collected and evaporated. The residue was purified by
column chromatography on silica gel with a 7:3:0.4 mixture
of CHCl3, MeOH, and H2O to give the pure 27 (3.3 mg,
25% in two steps) as a green oil. [a]2D5 C15.6 (c 0.22,
4.1.17. Compound 23. To a solution of the crude 19
(14.7 mg) in 1,4-dioxane (2 mL)–H2O (2 mL) were added
22 (26.9 mg, 46 mmol) and triethylamine (10 mL, 72 mmol)
at rt. After stirring for 4 h at rt, the reaction mixture was
dissolved in AcOEt and water, and extracted three times
with water. The water layers were combined and evaporated
to half to original volume. The water layer was poured into
Diaion HP-20. The resin was washed with water, and then
eluted with MeOH. The MeOH fractions were then
collected and evaporated. The residue was purified by
column chromatography on silica gel with a 7:3.5:0.6
mixture of CHCl3, MeOH, and H2O to give the pure 23
(4.7 mg, 14% in two steps) as an orange oil. [a]2D5 C10.8
(c 0.13, MeOH–H2OZ4:1); 1H NMR (400 MHz, CD3OD):
dZ1.35 (m, 2H), 1.58 (m, 4H), 2.15 (t, JZ7.3 Hz, 2H),
3.33–3.43 (m, 4H), 3.46 (dd, JZ10.3, 3.0 Hz, 1H), 3.52–
3.75 (m, 9H), 3.81 (m, 2H), 3.89 (d, JZ2.4 Hz, 1H), 4.17
(t, JZ6.6 Hz, 1H), 4.20 (d, JZ7.3 Hz, 1H), 4.86 (d, JZ
3.2 Hz, 1H), 6.48 (dd, JZ9.1, 2.5 Hz, 2H), 6.58 (d, JZ
2.5 Hz, 2H), 6.66 (d, JZ9.1 Hz, 2H), 7.21 (d, JZ8.1 Hz,
1H), 8.04 (dd, JZ8.1, 1.7 Hz, 1H), 8.33 (d, JZ1.7 Hz, 1H);
HRMS (ESI-TOF): calcd for C41H49N2O18H (MCH)C:
857.2975. Found: 857.2944.
1
MeOH–H2OZ4:1); H NMR (400 MHz, CD3OD–D2OZ
4:1): dZ1.13 (m, 2H), 1.31 (m, 4H), 2.02 (t, JZ7.6 Hz,
2H), 2.84 (t, JZ7.1 Hz, 2H), 2.87 (s, 6H), 3.43 (m, 3H),
3.52–3.74 (m, 10H), 3.79–3.94 (m 6H), 3.99 (d, JZ2.7 Hz,
1H), 4.28 (t, JZ6.3 Hz, 1H), 4.35 (d, JZ8.8 Hz, 1H), 4.44
(d, JZ7.5 Hz, 1H), 7.31 (d, JZ7.3 Hz, 1H), 7.62 (m, 2H),
8.17 (dd, JZ7.3, 1.2 Hz, 1H), 8.30 (d, JZ9.8 Hz, 1H), 8.53
(d, JZ8.5 Hz, 1H); 13C NMR (150 MHz, CD3OD–D2OZ
4:1): dZ26.11, 26.81, 29.97, 36.69, 40.38, 43.62, 45.90,
61.49, 62.22, 69.61, 70.22, 70.63, 70.90, 71.19, 72.50,
74.14, 74.50, 76.00, 76.25, 76.56, 79.33, 80.62, 102.24,
103.95, 104.99, 116.59, 120.38, 124.51, 129.31, 130.18,
130.68, 130.84, 131.12, 136.51, 152.83, 176.69; HRMS
(ESI-TOF): calcd for C37H57N3O19SH (MCH)C:
894.3536. Found: 894.3524.
4.1.22. Compound 28. To a solution of the crude compound
26 (9.4 mg) in 1,4-dioxane (3 mL)–H2O (3 mL) were added
22 (10.0 mg, 17 mmol) and triethylamine (12 mL, 86 mmol)
at rt. After stirring for 23 h at rt, the reaction mixture was
dissolved in AcOEt and water, and extracted three times
with water. The water layers were combined and evaporated
to half to original volume. The water layer was poured into
Diaion HP-20. The resin was washed with water, and then
eluted with MeOH. The MeOH fractions were then
collected and evaporated. The residue was purified by
column chromatography on silica gel with a 6:4:0.9 mixture
of CHCl3, MeOH, and H2O to give the pure 28 (6.1 mg,
35% in two steps) as an orange oil. [a]2D5 C26.8 (c 0.33,
MeOH–H2OZ4:1); 1H NMR (400 MHz, CD3OD): dZ1.44
(m, 2H), 1.68 (m, 4H), 2.24 (t, JZ7.3 Hz, 1H), 3.42–3.59
(m, 9H), 3.64–3.97 (m, 14H), 4.24 (t, JZ6.3 Hz, 1H), 4.31
(d, JZ8.6 Hz, 1H), 4.40 (m, 1H), 4.94 (d, JZ3.9 Hz, 1H),
6.57 (d, JZ8.8 Hz, 2H), 6.68 (s, 2H), 6.73 (d, JZ8.8 Hz,
2H), 7.31 (d, JZ8.1 Hz, 1H), 8.14 (d, JZ8.1 Hz, 1H), 8.43
4.1.18. Compound 24. A solution of 15 (46.2 mg, 51 mmol)
was treated with ozone at K78 8C by bubbling until the
solution remained blue. The reaction mixture was evacuated
with air for 5 min to remove the ozone, and then dimethyl
sulfide (0.2 mL) was added at K78 8C to the reaction
mixture. The resulting solution was allowed to warm to rt,
and then evaporated to afford a crude 24 (44.4 mg). The
crude 24 was used in the next step without further
purification due to its lability.
4.1.19. Compound 25. Hydroxylamine hydrochloric acid
salt (4.9 mg, 71 mmol) was added at rt to a solution of 24
(44.4 mg, 49 mmol) in MeOH (60 mL)–H2O (8 mL). After
stirring for 2 h at rt, MeOH was evaporated. The reaction
mixture was quenched with water, and extracted three times
with AcOEt. The AcOEt layers were combined, washed
with brine, dried over anhydrous Na2SO4, and evaporated.
The residue was purified by column chromatography on
silica gel with a 10:1 mixture of CHCl3 and MeOH to give
an inseparable mixture of E- and Z-isomers 25 (31.6 mg,
67% in two steps) as a colorless amorphous solid.
1
(s, 1H); H NMR (400 MHz, CD3OD–D2OZ4:1): dZ1.43
(m, 2H), 1.67 (m, 4H), 2.26 (t, JZ7.3 Hz, 2H), 3.44
(m, 5H), 3.54–3.73 (m, 9H), 3.81–3.98 (m, 9H), 4.28 (t,
JZ5.9 Hz, 1H), 4.35 (d, JZ7.8 Hz, 1H), 4.44 (d, JZ
7.3 Hz, 1H), 6.62 (d, JZ8.7 Hz, 2H), 7.75 (m, 4H), 7.31
(d, JZ8.1 Hz, 1H), 8.13 (d, JZ8.1 Hz, 1H), 8.40 (s, 1H);

6526
T. Miura et al. / Tetrahedron 61 (2005) 6518–6526
MALDI-TOF MS found: m/z [MCH]C 1019.9. Calcd for
C47H59N2O23 [MCH]C 1019.4. Found: m/z [MCNa]C
1041.9. Calcd for C47H58N2O23Na [MCNa]C 1041.3.
Found: m/z [MCK]C 1057.7. Calcd for C47H58N2O23K
[MCK]C 1057.3.
7. (a) Wipf, P.; Reeves, J. T. Tetrahedron Lett. 1999, 40, 5139.
(b) Studer, A.; Curran, D. P. Tetrahedron 1997, 53, 6681.
(c) Wipf, P.; Reeves, J. T.; Balachandran, R.; Giuliano, K. A.;
Hamel, E.; Day, B. W. J. Am. Chem. Soc. 2000, 122, 9391.
¨
(d) Rover, S.; Wipf, P. Tetrahedron Lett. 1999, 40, 5667.
(e) Wipf, P.; Reeves, J. T. Tetrahedron Lett. 1999, 40, 4649.
(f) Curran, D. P.; Ferritto, R.; Hua, Y. Tetrahedron Lett.
1998, 39, 4937. (g) Curran, D. P.; Amatore, M.; Guthrie, D.;
Campbell, M.; Go, E.; Luo, Z. J. Org. Chem. 2003, 68, 4643.
(h) Schwinn, D.; Bannwarth, W. Helv. Chim. Acta 2002, 85,
255. (i) Filippov, D. V.; Zoelen, D. J.; Oldfield, S. P.; Marel,
G. A.; Overkleeft, H. S.; Drijfhout, J. W.; Boom, J. H.
Tetrahedron Lett. 2002, 43, 7809. (j) Pardo, J.; Cobas, A.;
Acknowledgements
This work was partly supported by a Grant-in-Aid for
Young Scientists (B) (No. 16790026) from the Japan
Society for the Promotion of Science and a grant for Hi-
Tech Research from Tokai University. This work was
performed through the Noguchi Fluorous Project by our
institute.
´
Guitlan, E.; Castedo, L. Org. Lett. 2001, 3, 3711. (k) Luo, Z.;
Williams, J.; Read, R. W.; Curran, D. P. J. Org. Chem. 2001,
66, 4261.
8. (a) Miura, T.; Hirose, Y.; Ohmae, M.; Inazu, T. Org. Lett.
2001, 3, 3947. (b) Miura, T.; Goto, K.; Waragai, H.;
Matsumoto, H.; Hirose, Y.; Ohmae, M.; Ishida, H.-K.; Satoh,
A.; Inazu, T. J. Org. Chem. 2004, 69, 5348. (c) Miura, T.;
Satoh, A.; Goto, K.; Murakami, Y.; Imai, N.; Inazu, T.
Tetrahedron: Asymmetry 2005, 16, 3.
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