Synthesis of α-series ganglioside GM1α containing C20-sphingosine

Article abstract of DOI:10.1016/j.carres.2004.11.023

A synthesis of α-series ganglioside GM1α (III ⁶Neu5AcGgOse₄Cer) containing C20-sphingosine(d20:1) is described. Glycosylation of 2-(trimethylsilyl)ethyl 2,3,6-tri-O-benzyl-β-d- galactopyranosyl-(1→4)-2,3,6-tri-O-benzyl-β-d-glucopyranoside with the glucosamine donor ethyl 3-O-acetyl-2-deoxy-4,6-O-[(4-methoxyphenyl)methylene]- 2-phthalimido-1-thio-β-d-glucopyranoside furnished a β-(1→4)- linked trisaccharide. Reductive cleavage of the p-methoxybenzylidene group followed by intramolecular inversion of its triflate afforded the desired trisaccharide, which was transformed into a trisaccharide acceptor via removal of the phthaloyl and O-acetyl groups followed by N-acetylation. A tetrasaccharide acceptor was obtained by glycosylation of the trisaccharide acceptor with dodecyl 2,3,4,6-tetra-O-benzoyl-1-thio-β-d-galactopyranoside, followed by removal of the p-methoxybenzyl group. Coupling of the tetrasaccharide acceptor with ethyl (methyl 4,7,8,9-tetra-O-acetyl-3,5-dideoxy- 1-thio-5-trichloroacetamido-d-glycero-d-galacto-2-nonulopyranosid)onate and subsequent radical reduction gave the desired GM1α saccharide derivative, which was coupled with (2S,3R,4E)-2-azido-3-O-benzoyl-4-eicosene-1,3-diol after conversion into the imidate.

Full text of DOI:10.1016/j.carres.2004.11.023

Carbohydrate
RESEARCH
Carbohydrate Research 340 (2005) 211–220
Synthesis of a-series ganglioside GM1a containing C20-sphingosine
Yoichi Takeda and Shigeomi Horito*
Department of Biological Science & Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan
Received 22 July 2004; received in revised form 12 November 2004; accepted 26 November 2004
Abstract—A synthesis of a-series ganglioside GM1a (III6Neu5AcGgOse4Cer) containing C20-sphingosine(d20:1) is described. Gly-
cosylation of 2-(trimethylsilyl)ethyl 2,3,6-tri-O-benzyl-b-D-galactopyranosyl-(1!4)-2,3,6-tri-O-benzyl-b-D-glucopyranoside with the
glucosamine donor ethyl 3-O-acetyl-2-deoxy-4,6-O-[(4-methoxyphenyl)methylene]-2-phthalimido-1-thio-b-D-glucopyranoside fur-
nished a b-(1!4)-linked trisaccharide. Reductive cleavage of the p-methoxybenzylidene group followed by intramolecular inversion
of its triflate afforded the desired trisaccharide, which was transformed into a trisaccharide acceptor via removal of the phthaloyl
and O-acetyl groups followed by N-acetylation. A tetrasaccharide acceptor was obtained by glycosylation of the trisaccharide accep-
tor with dodecyl 2,3,4,6-tetra-O-benzoyl-1-thio-b-D-galactopyranoside, followed by removal of the p-methoxybenzyl group. Cou-
pling of the tetrasaccharide acceptor with ethyl (methyl 4,7,8,9-tetra-O-acetyl-3,5-dideoxy-1-thio-5-trichloroacetamido-D-glycero-
D-galacto-2-nonulopyranosid)onate and subsequent radical reduction gave the desired GM1a saccharide derivative, which was
coupled with (2S,3R,4E)-2-azido-3-O-benzoyl-4-eicosene-1,3-diol after conversion into the imidate.
Ó 2004 Elsevier Ltd. All rights reserved.
Keywords: a-Series gangliosides; Cholinergic-specific ganglioside antigen; Chol-1; GM1a; Intramolecular inversion; a-Sialylation; N-Trichloroacet-
ylneuraminic acid; Eicosene
1. Introduction
GD1aa and GT1ba; each ganglioside was composed
of various ceramides; two major components were
sphingosines (d18:1 and d20:1).6 One interesting fact is
that the sphingosine d20:1 was not found in brain sphin-
gomyelin but only brain gangliosides. These variations
in ceramide structure are generally expressed in a cell-
type-specific and developmentally regulated manner,
implying that they have some specific purposes. For elu-
cidating the specific purposes, it is necessary to examine
the physicochemical characters of some gangliosides
having a different ceramide. The synthesis of GM1a
containing sphingosine d18:1 and fatty acid C18:0 has
been reported.7 We present herein the synthesis of
GM1a comprised of sphingosine d20:1 and octadeca-
noic acid C18:0.
Gangliosides are ubiquitously located in vertebrate cell
and are particularly abundant in the nervous system.
However, a-series gangliosides containing a common
sialyl a(2!6)N-acetylgalactosamine residue are exclu-
sively localized on cholinergic neurons1 and are well
known as a cholinergic-specific ganglioside antigen
(Chol-1).2 There have been several reports suggesting
putative roles of a-series gangliosides in the nervous sys-
tem such as the specific association of GT1aa and
GQ1ba with cholinergic neuron of rat brain,3 the spe-
cific cell adhesion of GQ1ba mediated by myelin-associ-
ated glycoprotein4 and the potent neuritogenic activity
of GT1aa towards cultured neuronal cells (Neuro-
2A).5 Three extremely minor but novel Chol-1 antigens
isolated from bovine brain were identified as GM1a,
2. Results and discussion
Because gangliosides having many variations in
ceramide structure would be in demand, we decided to
*
Corresponding author. Fax: +81 4 7125 1841; e-mail: shorito@
rs.noda.tus.ac.jp
0008-6215/$ - see front matter Ó 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2004.11.023

212
Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
amine moiety. O-Deacetylation and cleavage of phthal-
OH OBn
O
OBn
O
O
imide group with hydrazine monohydrate followed by
N-acetylation yielded the trisaccharide acceptor 6
(89%). Condensation of 6 with the galactosyl donor 7
obtained from dodecyl 2,3,4,6-tetra-O-acetyl-1-thio-b-
D-galactopyranoside10 by deacetylation and benzoyl-
ation afforded the b-linked tetrasaccharide 8 (58%).
The signal at 4.92 (d, J1,2 7.9 Hz, H-1IV) ppm in the
1H NMR spectrum of 8 proves the b-configuration of
the newly synthesized linkage. Removal of the p-meth-
oxybenzyl group with ammonium cerium(IV) nitrate
produced the tetrasaccharide acceptor 9 in 91% yield.
Our earlier experience with a sialyl donor having 5-N-
trichloroacetyl group had shown that the sialyl donor
ethyl (methyl 4,7,8,9-tetra-O-acetyl-3,5-dideoxy-1-thio-
5-trichloroacetamido-D-glycero-D-galacto-2-nonulopyr-
anosid)onate (10) gave preferentially the a-glycoside in
CH2Cl2 or CH2Cl2–Et2O.11 Consequently, when we
started to couple the donor 10 with the acceptor 9,
CH2Cl2 was tried as a solvent. The amount of TfOH re-
quired for the beginning of the reaction was unsettled
because of low solubility of TfOH in CH2Cl2, though
the a-linked pentasaccharide 11 was obtained in 50%
yield. This problem was solved by the use of CH3CN
in which TfOH is completely soluble. Sialylation of 9
with the donor 10 by use of NIS–TfOH in CH3CN affor-
ded stereoselectively the a-linked pentasaccharide 11 in
60% yield (Scheme 2). Radical reduction with t-BuSnH
and AIBN gave the sialopentasacccharide 12 (96%).
The signals at 4.82 (H-4V), 2.54 (H-3eqV) and 1.85 (H-
3axV) ppm and the value of d jH-9aV–H-9bVj =
0.22 ppm in the 1H NMR spectrum of 12 prove the
a-configuration of the newly synthesized linkage.
Removal of the benzyl group with NaBrO3 and
Na2S2O4 followed by benzoylation afforded the GM1a
oligosaccharide derivative 13 (59%).
BnO
O
O
MeO
AcO
SEt
BnO
OSE
O
OBn
NPhth
2
1
OBn
a
R2O
O
R1O
AcO
O
OBn
O
PhthN
OBn
OSE
BnO
O
BnO
O
OBn
3 R1, R2 = CHC6H4OMe-p (91%)
4 R1 = H, R2 = PMB (88%)
OBn
b
d
R2O
HO
c
OPMB
O
O
OBn
R1
BnO
OBn
O
BnO
O
OSE
O
OBn
OBn
5 R1 = NPhth, R2 = Ac (83%)
6 R1 = NHAc, R2 = H (89%)
BzO OBz
O
e
SLau
BzO
OBz
7
HO
O
OR
BzO OBz
O
O
BzO
O
OBn
O
OBz
AcHN
BnO
OBn
BnO
O
OSE
O
OBn
8 R = PMB (58%)
9 R = H (91%)
f
OBn
˚
Scheme 1. Reagents and conditions: (a) NIS–TfOH, MS 4 A, CH2Cl2,
˚
ꢀ40 °C; (b) NaB(CN)H3, TFA, MS 4 A, DMF, 0 °C to rt; (c) Tf2O,
CH2Cl2–pyridine, 0 °C to rt; DMF, 60 °C; (d) NH2NH2ÆH2O, EtOH,
˚
reflux; Ac2O, pyridine, rt; (e) NIS–TfOH, MS 4 A, CH2Cl2, 0 °C; (f)
CAN, CH3CN, rt. SE: 2-(trimethylsilyl)ethyl; PMB: p-methoxybenzyl;
NPhth: phthalimide; Lau: laulyl.
use a glucosamine derivative as a starting material
for the galactosamine moiety. As shown in Scheme 1,
condensation of the lactose acceptor 2-(trimethylsilyl)-
ethyl 2,3,6-tri-O-benzyl-b-D-galactopyranosyl-(1!4)-
2,3,6-tri-O-benzyl-b-D-glucopyranoside (2)8 with the
glucosamine donor ethyl 3-O-acetyl-2-deoxy-4,6-O-[(4-
methoxyphenyl)methylene]-2-phthalimido-1-thio-b-D-
glucopyranoside (1)9 in CH2Cl2 in the presence of
N-iodosuccinimide (NIS)-TfOH and molecular sieves
Removal of the 2-(trimethylsilyl)ethyl group and sub-
sequent trichloroacetimidate formation gave the penta-
saccharide donor; its condensation with (2S,3R,4E)-2-
azido-3-O-benzoyl-4-eicosene-1,3-diol (14)12 afforded
the b-linked glycosyl sphingosine derivative 15 (36%).
The signal at 4.54 (d, 1H, J1,2 7.6 Hz, H-1I) ppm in
the 1H NMR spectrum of 15 proves the b-configuration
of the newly synthesized linkage. Reduction of the azi-
do group of 15 with Zn powder and subsequent cou-
pling of the amino group with octadecanoic acid
using N-(3-dimethylaminopropyl)-N0-ethylcarbodiimide
hydrochloride gave the ganglioside derivative 16 in
74% yield.
One-pot deprotection of 16 through methanolysis and
saponification of the methyl ester group gave the title
ganglioside GM1a 17 comprised of sphingosine d20:1
and fatty acid C18:0 in 76% yield. The signals at 4.63
(d, 1H, J1,2 8.5 Hz, H-1IV), 4.37 (d, 1H, J1,2 7.7 Hz, H-
1II), 4.35 (d, 1H, J1,2 7.6 Hz, H-1III), 4.31 (d, 1H, J1,2
7.7 Hz, H-1I), 2.35 (H-3eqV), 1.78 (H-3axV) ppm in the
˚
(4 A) afforded the b-linked trisaccharide 3 in 91% yield.
1
The signal at 5.48 (d, J1,2 8.2 Hz, H-1III) ppm in the H
NMR spectrum of 3 proves the b-configuration of the
newly synthesized linkage. Reductive cleavage of the
p-methoxybenzylidene group of 3 by NaB(CN)H3 and
CF3CO2H offered the 6-O-p-methoxybenzyl derivative
4 (88%). Intramolecular inversion by the acetyl group
of 4 via the triflate yielded the target intermediate 5
having galacto-configuration in a high yield (83%).
1
The signal 5.49 (d, J3,4 3.4 Hz, H-4III) ppm in the H
NMR spectrum of 5 proves the formation of galactos-

Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
213
AcO
COOMe
SEt
AcO
9
O
TCAHN
AcO
OAc
10
AcO
COOMe
O
AcO
a
R3HN
O
AcO
OAc
BzO OBz R2O
O
O
O
OR1
O
O
BzO
OR1
OBz
AcHN
R1O
R1O
O
OSE
OR1
O
OR1
11 R1 = Bn, R2 = H, R3 = TCA (60%)
12 R1 = Bn, R2 = H, R3 = Ac (96%)
13 R1 = Bz, R2 = Bz, R3 = Ac (59%)
b
c
OBz
HO
AcO
(CH2)14CH3
COOMe
O
AcO
d
N3
14
O
AcHN
AcO
OAc
BzO OBz BzO
O
O
O
BzO
O
OBz
O
OBz
OBz
AcHN
OBz
BzO
O
BzO
O
(CH2)14CH3
O
OBz
15 R = N3 (36%)
16 R = NHCO(CH2)16CH3 (74%)
R
e
OBz
HO
COOH
f
HO
O
AcHN
HO
O
OH
HO OH
HO
O
O
O
HO
O
OH
OH
OH
AcHN
OH
O
HO
O
HO
O
(CH2)14CH3
NHCO(CH2)16CH3
17 (76%)
O
OH
OH
˚
Scheme 2. Reagents and conditions: (a) NIS–TfOH, MS 3 A, CH3CN, ꢀ40 °C; (b) BuSnH, AIBN, C6H6, reflux; (c) NaBrO3, Na2S2O4, AcOEt–H2O,
˚
rt; BzCl, pyridine; (d) TFA, CH2Cl2, 0 °C; CCl3CN, DBU, CH2Cl2, rt; BF3ÆEt2O, MS 4 A, CH2Cl2, 0 °C; (e) Zn powder, AcOH–CH2Cl2, rt;
CH3(CH2)16COOH, Me2HN(CH2)3N@C@NEtCl, CH2Cl2, rt; (f) MeONa, MeOH, rt; H2O, rt. TCA: CCl3CO.
1H NMR spectrum of 17 clearly prove the desired ano-
meric configuration of the synthesized linkages.
mental analyses were performed on a Perkin-Elmer 2400
II elemental analyzer. All reactions were monitored by
TLC on aluminium sheets coated with silica gel 60F254
(0.2 mm thickness, E. Merck, Darmstadt, Germany).
Column chromatography was carried out using Wako
gel C-300E (Wako). Solvent extracts were dried with
anhydrous MgSO4 unless otherwise specified.
3. Experimental
3.1. General procedures
Optical rotations were measured at 25 °C with a JASCO
3.2. 2-(Trimethylsilyl)ethyl O-[3-O-acetyl-2-deoxy-4,6-O-
[(4-methoxyphenyl)methylene]-2-phthalimido-b-D-glucopyr-
anosyl]-(1!4)-O-(2,3,6-tri-O-benzyl-b-D-galactopyrano-
syl)-(1!4)-2,3,6-tri-O-benzyl-b-D-glucopyranoside (3)
To a solution of 28 (428 mg, 0.44 mmol) and 19
(380mg, 0.74 mmol) in anhyd CH2Cl2 (5 mL) were
1
DIP-370 polarimeter. H NMR and 13C NMR spectra
were recorded on a Bruker DRX-600 spectrometer at
600 and 125 MHz, respectively. In CDCl3, the dH-value
is relative to internal Me4Si and the dC-value is refer-
enced to the solvent [dC (CDCl3) 77.0]. Assignments
1
were aided by COSY, TOCSY and H–13C correlation
˚
added molecular sieves 4 A (480 mg) and NIS
(343 mg, 1.52 mmol), then cooled to ꢀ40 °C. To the
spectroscopy. High-resolution mass spectra were re-
corded on a JEOL JMS-700 under FAB conditions. Ele-

214
Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
mixture was added TfOH (10 lL, 0.11 mmol), and stir-
ring was continued for 40 min at ꢀ40 °C. The reaction
was quenched with Et3N (50.0 lL). The precipitate was
filtered off and washed with CH2Cl2. The filtrate and
washings were combined, and the solution was succes-
sively washed with satd aq NaHCO3, M Na2S2O3 and
satd aq NaCl, dried and concentrated. Purification by
flash column chromatography (1:4 EtOAc–hexane)
gave 3 (567 mg, 91%) as an amorphous mass. [a]D
+10.5 (c 1.00, CHCl3); 1H NMR (CDCl3): d 7.85–
6.84 (m, 38H, 6Ph, NPhth, p-MeOC6H4), 6.11 (t, 1H,
H-3III), 5.52 (s, 1H, p-MeOC6H4CH), 5.48 (d, 1H,
J1,2 8.2 Hz, H-1III), 5.10 (d, 1H, Jgem 10.2 Hz, CHHPh),
4.89 (d, 1H, Jgem 10.8 Hz, CHHPh), 4.79 (d, 1H, Jgem
10.2 Hz, CHHPh), 4.77 (d, 1H, Jgem 10.8 Hz, CHHPh),
4.46 (d, 1H, Jgem 11.8 Hz, CHHPh), 4.40 (d, 1H, Jgem
12.1 Hz, CHHPh), 4.32 (d, 1H, J1,2 8.8 Hz, H-1I),
4.33 (d, 1H, J1,2 8.2 Hz, H-2III), 4.22 (d, 1H, J1,2
8.8 Hz, H-1II), 4.19 (H-5III), 4.17 (H-6aIII), 3.97 (m,
1H, CHHCH2Si), 3.82 (t, 1H, J3,4 9.6 Hz, H-4I), 3.80
(OCH3), 3.75 (H-6bIII), 3.72 (H-4III), 3.71 (H-4II),
3.61 (H-6aI), 3.56 (t, 1H, J2,3 9.5 Hz, J3,4 9.6 Hz, H-
3I), 3.55 (H-6aII, H-6bI), 3.54 (m, 1H, CHHCH2Si),
3.33 (t, 1H, J1,2 8.8 Hz, J2,3 9.5 Hz, H-2I), 3.32 (H-
6bII), 3.27 (m, 1H, H-5I), 3.12 (H-5II), 3.10 (t, 1H,
J2,3 8.8 Hz, H-3II), 3.02 (t, 1H, J2,3 8.8 Hz, H-2II),
1.88 (s, 3H, COCH3), 1.01 (m, 2H, CH2CH2Si), 0.00
(s, 9H, 3SiCH3). 13C NMR (CDCl3): d 190.80, 170.07,
160.11, 139.49, 138.90, 138.34, 138.27, 137.83, 131.94,
129.34, 128.89, 128.62, 128.47, 128.35, 128.30, 128.26,
128.20, 128.12, 128.05, 127.86, 127.76, 127.72, 127.63,
127.52, 127.40, 127.29, 127.16, 127.04, 114.23, 113.56,
103.08 (C-1I), 101.87 (C-1II), 101.51 (CHC6H4OMe-p),
99.67 (C-1III), 82.81 (C-3I), 81.71 (C-2I), 80.41 (C-3II),
80.08 (C-2II), 79.35 (C-4II), 76.41 (C-4I), 75.63, 75.01
(C-5I), 74.93, 74.69, 73.18, 73.02, 72.96, 72.66 (C-5III),
72.30 (C-5II), 69.25, 68.59, 68.18 (C-6I), 68.06, 67.25
(OCH2CH2Si), 65.50 (C-4III), 60.34 (C-6II), 55.52,
55.32, 55.23 (OCH3), 20.66 (COCH3), 18.38 (CH2Si),
14.14, ꢀ1.45 (SiCH3). Anal. Calcd for C83H91NO19Si:
C, 69.48; H, 6.39; N, 0.98. Found: C, 69.55; H, 6.56;
N, 1.04.
sively washed with satd aq NaCl, dried and concen-
trated. Purification by flash column chromatography
(1:2 EtOAc–hexane) gave 4 (637 mg, 88%) as an amor-
phous mass. [a]D +13.9 (c 1.00, CHCl3); 1H NMR
(CDCl3):
d 7.65–6.82 (m, 38H, 6Ph, NPhth, p-
MeOC6H4), 5.94 (dd, 1H, J2,3 11.0 Hz, J3,4 9.0 Hz, H-
3III), 5.39 (d, 1H, J1,2 8.3 Hz, H-1III), 5.08 (d, 1H, Jgem
9.8 Hz, CHHPh), 4.89 (d, 1H, Jgem 11.0 Hz, CHHPh),
4.79 (d, 1H, Jgem 11.0 Hz, CHHPh), 4.76 (d, 1H, Jgem
9.8 Hz, CHHPh), 4.46 (d, 1H, Jgem 12.1 Hz, CHHPh),
4.45 (d, 1H, Jgem 11.1 Hz, CHHPh), 4.44 (d, 1H, Jgem
12.1 Hz, CHHPh), 4.43 (d, 1H, Jgem 11.1 Hz, CHHPh),
4.34 (d, 1H, J1,2 7.8 Hz, H-1I), 4.33 (t, 1H, J1,2 8.3 Hz,
J2,3 11.0 Hz, H-2III), 4.20 (d, 1H, J1,2 7.6 Hz, H-1II),
4.14 (d, 1H, Jgem 10.8 Hz, p-MeOC6H4CHH), 3.98 (m,
1H, CHHCH2Si), 3.90 (t, 1H, J3,4 9.0 Hz, H-4III), 3.82
(t, 1H, J3,4 9.3 Hz, J4,5 9.6 Hz, H-4I), 3.76 (H-6aIII),
3.76 (s, 3H, OCH3), 3.73 (d, 1H, J3,4 2.3 Hz, H-4II),
3.69 (H-6bIII), 3.69 (H-5III), 3.68 (H-6aII), 3.62 (H-6aI),
3.59 (H-6bI), 3.55 (m, 1H, CHHCH2Si), 3.53 (t, 1H,
J2,3 9.1 Hz, J3,4 9.3 Hz, H-3I), 3.47 (d, 1H, Jgem
10.8 Hz, p-MeOC6H4CHH), 3.35 (dd, 1H, J5,6b 5.1 Hz,
H-6bII), 3.31 (dd, 1H, J1,2 7.8 Hz, J2,3 9.1 Hz, H-2I),
3.27 (ddd, 1H, J4,5 9.6 Hz, H-5I), 3.13 (dd, 1H, J5,6b
5.1 Hz, H-5II), 3.10 (dd, 1H, J2,3 9.6 Hz, J3,4 2.3 Hz,
H-3II), 3.01 (dd, 1H, J1,2 7.6 Hz, J2,3 9.6 Hz, H-2II),
1.91 (s, 3H, COCH3), 1.01 (m, 2H, CH2CH2Si), 0.00
(s, 9H, 3SiCH3). 13C NMR (CDCl3): d 171.31, 159.28,
139.07, 138.93, 138.66, 138.29, 138.26, 137.95, 129.54,
129.33, 129.25, 128.38, 128.20, 128.11, 128.05, 127.96,
127.84, 127.67, 127.41, 127.36, 127.31, 127.26, 113.83,
103.08 (C-1I), 101.81 (C-1II), 99.19 (C-1III), 82.78 (C-
3I), 81.74 (C-2I), 80.48 (C-3II), 80.10 (C-2II), 76.26 (C-
4I), 75.93, 75.05, 74.96 (C-5I), 74.91 (C-4II), 74.65,
73.38, 73.21 (C-5III), 72.98, 72.95 (C-3III), 72.86, 72.73
(C-5II), 72.44, 71.51 (C-4III), 69.68 (C-6III), 68.47 (C-
6II), 68.15 (C-6I), 67.25 (OCH2), 55.19 (OCH3), 54.58
(C-2III), 20.76 (COCH3), 18.38 (CH2Si), ꢀ1.46 (SiCH3).
Anal. Calcd for C83H93NO19Si: C, 69.39; H, 6.52; N,
0.97. Found: C, 69.27; H, 6.71; N, 0.88.
3.4. 2-(Trimethylsilyl)ethyl O-[4-O-acetyl-2-deoxy-6-O-
[(4-methoxyphenyl)methyl]-2-phthalimido-b-D-galacto-
pyranosyl]-(1!4)-O-(2,3,6-tri-O-benzyl-b-D-galactopyr-
anosyl)-(1!4)-2,3,6-tri-O-benzyl-b-D-glucopyranoside (5)
3.3. 2-(Trimethylsilyl)ethyl O-[3-O-acetyl-2-deoxy-6-O-
[(4-methoxyphenyl)methyl]-2-phthalimido-b-D-glucopyr-
anosyl]-(1!4)-O-(2,3,6-tri-O-benzyl-b-D-galactopyrano-
syl)-(1!4)-2,3,6-tri-O-benzyl-b-D-glucopyranoside (4)
A solution of 4 (733 mg, 0.51 mmol) in anhyd CH2Cl2
(16 mL) and anhyd pyridine (3.1 mL) was cooled to
0 °C and Tf2O (257 lL, 1.5 mmol) added dropwise.
The mixture was stirred at rt for 1 h, then diluted with
anhyd CH2Cl2, successively washed with 2 M HCl and
satd aq NaCl, dried and the residue was co-evaporated
with toluene (three times) at rt. The residue was dis-
solved in anhyd DMF (100 mL). The solution was
heated to 60 °C and kept for 1 h and concentrated. Puri-
fication by flash column chromatography (1:3 EtOAc–
A solution of 3 (721 mg, 0.50 mmol) and molecular
˚
sieves 4 A AW300 (2.7 g) in anhyd DMF (4.2 mL) was
cooled to 0 °C and NaB(CN)H3 (333 mg, 5.0 mmol)
was added. A solution of TFA (135 lL) in anhyd
DMF (7.9 mL) was then added dropwise and the mix-
ture was allowed to stir at rt for 24 h. The reaction mix-
ture was poured into cold satd aq NaHCO3 and
extracted with CH2Cl2. The organic layer was succes-

Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
215
hexane) gave 5 (606 mg, 83%) as an amorphous mass.
1
trated. Purification by flash column chromatography
(5:7 EtOAc–hexane) gave 6 (553 mg, 89%) as an amor-
phous mass. [a]D +8.2 (c 1.00, CHCl3); 1H NMR
(CDCl3): d 7.41–7.17 (m, 34H, 6Ph, p-MeOC6H4), 4.90
(d, 1H, Jgem 10.4 Hz, CHHPh), 4.89 (d, 1H, Jgem
10.8 Hz, CHHPh), 4.84 (d, 1H, Jgem 11.3 Hz, CHHPh),
4.74 (d, 1H, Jgem 10.8 Hz, CHHPh), 4.73 (d, 1H, Jgem
10.4 Hz, CHHPh), 4.72 (d, 1H, Jgem 9.8 Hz, CHHPh),
4.64 (d, 1H, Jgem 9.8 Hz, CHHPh), 4.57 (2 d, 2H, Jgem
11.3 Hz, Jgem 12.3 Hz, 2CHHPh), 4.43 (d, 1H, Jgem
11.9 Hz, CHHPh), 4.42 (d, 1H, J1,2 8.3 Hz, H-1II), 4.39
(d, 1H, J1,2 8.3 Hz, H-1III), 4.38 (d, 1H, Jgem 11.9 Hz,
CHHPh), 4.37 (d, 1H, Jgem 12.3 Hz, CHHPh), 4.36
(d, 1H, J1,2 7.9 Hz, H-1I), 4.34 (d, 1H, Jgem 12.3 Hz,
p-MeOC6H4CHH), 4.27 (d, 1H, Jgem 12.3 Hz, p-
MeOC6H4CHH), 4.01 (d, 1H, J3,4 2.3 Hz, H-4II), 3.98
(m, 1H, CHHCH2Si), 3.93 (t, 1H, J3,4 9.3 Hz, H-4I),
3.85 (d, 1H, J3,4 3.2 Hz, H-4III), 3.73–3.69 (m, 6H,
OCH3, H-6aI, H-6bI, H-2III), 3.55 (m, 1H, CHHCH2Si),
3.53 (t, 1H, J3,4 9.3 Hz, H-3I), 3.47 (m, 2H, H-2II, H-
3II), 3.42 (dd, 1H, J2,3 9.0 Hz, J3,4 3.2 Hz, H-3III), 3.36
(m, 2H, H-2I, H-5I), 1.50 (s, 3H, NCOCH3), 1.01 (m,
2H, CH2CH2Si), 0.00 (s, 9H, 3SiCH3). 13C NMR
(CDCl3): d 173.94, 159.16, 138.64, 138.59, 138.51,
138.22, 138.12, 136.24, 129.85, 129.38, 128.97, 128.82,
128.68, 128.42, 128.22, 128.18, 128.13, 127.95, 127.75,
127.57, 127.47, 127.40, 127.36, 113.73, 103.07 (C-1II),
102.72 (C-1III), 102.29 (C-1I), 82.39 (C-3I), 82.08,
81.87, 80.59, 76.46, 75.58 (C-4II), 75.49 (C-4I), 75.26,
74.99, 74.93, 74.86, 73.72, 73.25, 73.19, 73.10, 68.82,
68.55, 68.10, 67.31, 67.15, 56.27, 55.14 (OCH3), 22.14
(COCH3), 18.38 (CH2Si), ꢀ1.47 (SiCH3). Anal. Calcd
for C75H91NO17Si: C, 68.94; H, 7.02; N, 1.07. Found:
C, 68.91; H, 6.87; N, 1.01.
[a]D ꢀ2.3 (c 1.00, CHCl3); H NMR (CDCl3): d 7.85–
6.82 (m, 38H, 6Ph, NPhth, p-MeOC6H4), 5.49 (d, 1H,
J3,4 3.4 Hz, H-4III), 5.26 (d, 1H, J1,2 8.3 Hz, H-1III),
5.04 (d, 1H, Jgem 9.4 Hz, CHHPh), 4.98 (dd, 1H, J2,3
11.0 Hz, J3,4 3.4 Hz, H-3III), 4.91 (d, 1H, Jgem 11.1 Hz,
CHHPh), 4.82 (d, 1H, Jgem 11.1 Hz, CHHPh), 4.79 (d,
1H, Jgem 9.4 Hz, CHHPh), 4.45 (d, 1H, Jgem 12.3 Hz,
CHHPh), 4.43 (dd, 1H, J1,2 8.3 Hz, J2,3 11.0 Hz, H-
2III), 4.42 (d, 1H, Jgem 12.3 Hz, CHHPh), 4.40 (d, 1H,
Jgem 11.3 Hz, CHHPh), 4.34 (d, 1H, J1,2 7.9 Hz, H-1I),
4.30 (d, 1H, Jgem 11.3 Hz, CHHPh), 4.28 (d, 1H, Jgem
12.3 Hz, CHHPh), 4.27 (d, 1H, Jgem 12.3 Hz, CHHPh),
4.26 (d, 1H, Jgem 12.1 Hz, CHHPh), 4.16 (d, 1H, J1,2
7.6 Hz, H-1II), 4.14 (d, 1H, Jgem 11.0 Hz, p-
MeOC6H4CHH), 4.10 (d, 1H, Jgem 12.1 Hz, CHHPh),
3.98 (m, 1H, CHHCH2Si), 3.93 (m, 1H, J5,6b 5.7 Hz,
H-5III), 3.76 (s, 3H, OCH3), 3.76 (H-4I), 3.70 (d, 1H,
J3,4 2.6 Hz, H-4II), 3.64 (H-6aII), 3.60 (H-6aI, H-6bI),
3.54 (H-6aIII), 3.54 (m, 1H, CHHCH2Si), 3.53 (H-3I),
3.49 (m, 1H, J5,6b 5.7 Hz, H-6bIII), 3.41 (d, 1H, Jgem
11.0 Hz, p-MeOC6H4CHH), 3.31 (dd, 1H, J1,2 7.9 Hz,
H-2I), 3.28 (H-6bII), 3.27 (H-5I), 3.10 (m, 1H, H-5II),
3.08 (dd, 1H, J2,3 9.4 Hz, J3,4 2.6 Hz, H-3II), 3.03 (dd,
1H, J1,2 7.6 Hz, J2,3 9.4 Hz, H-2II), 2.12 (s, 3H, COCH3),
1.01 (m, 2H, CH2CH2Si), 0.00 (s, 9H, 3SiCH3). 13C
NMR (CDCl3): d 171.59, 168.74, 167.82, 159.24,
139.02, 138.95, 138.79, 138.29, 138.26, 137.96, 133.72,
133.45, 132.54, 131.77, 129.70, 129.57, 129.48, 128.28,
128.23, 128.18, 128.14, 128.10, 128.02, 127.95, 127.81,
127.64, 127.41, 127.37, 127.32, 127.25, 127.21, 127.16,
127.01, 123.16, 122.84, 113.76, 103.05 (C-1I), 102.02
(C-1II), 99.87 (C-1III), 82.70 (C-3I), 81.74 (C-2I), 80.42
(C-3II), 79.99 (C-2II), 76.57 (C-4I), 76.09, 75.28 (C-4II),
75.06 (C-5I), 74.96, 74.51, 73.06 (C-5II), 72.99, 72.94,
72.38, 71.56 (C-5III), 69.89 (C-4III), 68.65 (C-6II), 68.11
(C-6I), 67.44 (C-6III), 67.23, 66.74 (C-3III), 55.17
(OCH3), 54.47 (C-2III), 29.62, 21.00 (COCH3), 18.36
(CH2Si), ꢀ1.46 (SiCH3). Anal. Calcd for C83H93NO19Si:
C, 69.39; H, 6.52; N, 0.97. Found: C, 69.36; H, 6.69; N,
0.81.
3.6. Dodecyl 2,3,4,6-tetra-O-benzoyl-1-thio-b-D-galacto-
pyranoside (7)
Dodecyl 2,3,4,6-tetra-O-acetyl-1-thio-b-D-galactopyrano-
side10 (10.4 g, 19.5 mmol) was suspended in a solution of
anhyd MeOH (16 mL), anhyd CH2Cl2 (24 mL) and M
NaOMe (3.7 mL), the mixture was stirred at rt for 4 h.
This solution was then treated with ion-exchange resin
Amberlite IR-120 (H+) for about 5 min. The resin was
filtered off, and the solvent was removed under dimin-
ished pressure to quantitatively give a white solid
(7.1 g). To a stirred solution of the solid in anhyd pyr-
idine (60 mL) and anhyd CH2Cl2 (40 mL) was added a
solution of benzoyl chloride (12.9 mL, 111 mmol) drop-
wise at 0 °C. The reaction was stirred at rt for 6 h. The
reaction mixture was then diluted with anhyd CH2Cl2
(250 mL) and sequentially washed with 2 M HCl, satd
aq NaHCO3 and then satd aq NaCl. The organic layer
was dried and concentrated. Purification by flash col-
umn chromatography (1:10 EtOAc–hexane) gave 7
(13.8 g, 91%) as a colourless syrup. [a]D +60.0 (c 1.00,
3.5. 2-(Trimethylsilyl)ethyl O-[2-acetamido-2-deoxy-6-O-
[(4-methoxyphenyl)methyl]-b-D-galactopyranosyl]-(1!4)-
O-(2,3,6-tri-O-benzyl-b-D-galactopyranosyl)-(1!4)-
2,3,6-tri-O-benzyl-b-D-glucopyranoside (6)
A solution of 5 (686 mg, 0.48 mmol) in 95% EtOH
(14.3 mL) and hydrazine monohydrate (0.49 mL) heated
under reflux for 2 h. The mixture were evaporated and
co-evaporated several times with toluene to give a resi-
due, which was dissolved in MeOH (24 mL) and Ac2O
(3.5 mL), and stirred overnight at rt. After pyridine
(7 mL) was added, the mixture was concentrated and
solution of the residue in CH2Cl2 was successively
washed with 2 M HCl, satd aq NaCl, dried and concen-

216
Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
1
CHCl3); H NMR (CDCl3): d 8.01–7.15 (m, 20H, 4Ph),
5.97 (d, 1H, J3,4 3.4 Hz, H-4), 5.78 (t, 1H, J1,2 10.0 Hz,
J2,3 10.0 Hz, H-2), 5.58 (dd, 1H, J2,3 10.0 Hz, J3,4
3.4 Hz, H-3), 4.79 (d, 1H, J1,2 10.0 Hz, J2,3 10.0 Hz,
H-1), 4.60 (dd, 1H, H-6a), 4.34–4.29 (m, 2H, H-5, 6b),
2.79–2.66 (m, 2H, SCH2), 1.67–1.50 (m, 2H, SCH2CH2),
1.25–1.13 (m, 18H, 9CH2), 0.79 (t, 3H, CH3). 13C NMR
(CDCl3): d 165.95, 165.44, 165.38, 165.26 (4C@O),
133.60, 133.30, 133.27, 129.88, 129.73, 129.66, 129.16,
128.95, 128.79, 128.57, 128.53, 128.38, 128.31, 128.22,
84.30 (C-1), 74.77 (C-5), 72.53 (C-3), 68.15 (C-4),
68.00 (C-2), 62.08 (C-6), 31.85, 30.33, 29.71, 29.60,
29.57, 29.52, 29.48, 29.30, 29.08, 28.82, 22.64, 14.13.
Anal. Calcd for C46H52O9S: C, 70.74; H, 6.71; S, 4.11.
Found: C, 70.52; H, 6.90; S, 3.95.
102.41 (C-1II), 102.33 (C-1IV), 99.32 (C-1III), 82.98 (C-
3I), 81.89, 81.68, 80.13 (C-3III), 79.86, 76.48 (C-4I),
75.21, 75.18, 75.13, 74.93, 73.73, 73.25, 73.11, 73.03,
72.83, 72.36, 72.25 (C-4II), 71.55, 71.44 (C-3IV), 69.53
(C-2IV), 69.33, 69.09, 68.36 (C-4III), 68.00 (C-4IV),
67.29 (CH2CH2Si), 62.11, 55.19 (OCH3), 54.18 (C-2III),
22.77 (COCH3), 18.47 (CH2CH2Si), ꢀ1.42 (SiCH3).
Anal. Calcd for C109H117NO26Si: C, 69.45; H, 6.26; N,
0.74. Found: C, 69.73; H, 6.18; N, 0.65.
3.8. 2-(Trimethylsilyl)ethyl O-(2,3,4,6-tetra-O-benzoyl-b-
D-galactopyranosyl)-(1!3)-O-(2-acetamido-2-deoxy-b-D-
galactopyranosyl)-(1!4)-O-(2,3,6-tri-O-benzyl-b-D-
galactopyranosyl)-(1!4)-2,3,6-tri-O-benzyl-b-D-gluco-
pyranoside (9)
3.7. 2-(Trimethylsilyl)ethyl O-(2,3,4,6-tetra-O-benzoyl-b-
D-galactopyranosyl)-(1!3)-O-[2-acetamido-2-deoxy-6-
O-[(4-methoxyphenyl)methyl]-b-D-galactopyranosyl]-
(1!4)-O-(2,3,6-tri-O-benzyl-b-D-galactopyranosyl)-
(1!4)-2,3,6-tri-O-benzyl-b-D-glucopyranoside (8)
To a solution of 8 (488 mg, 0.26 mmol) in CH3CN
(4.6 mL) and water (1.1 mL) was added dropwise
ammonium cerium(IV) nitrate (426 mg, 0.78 mmol) in
CH3CN (6.8 mL) and the mixture was stirred for 1 h
at rt. The reaction mixture was diluted with EtOAc
and was successively washed with satd aq NaHCO3,
dried and concentrated. Purification by flash column
chromatography (1:1 EtOAc–hexane) gave 9 (415 mg,
91%) as an amorphous mass. [a]D +68.0 (c 1.00,
CHCl3); 1H NMR (CDCl3): d 7.85–6.82 (m, 50H,
10Ph), 5.94 (d, 1H, J3,4 3.6 Hz, H-4IV), 5.77 (dd, 1H,
J1,2 7.8 Hz, J2,3 10.5 Hz, H-2IV), 5.55 (dd, 1H, J2,3
10.5 Hz, J3,4 3.6 Hz, H-3IV), 5.18 (d, 1H, NH), 4.92
(d, 1H, J1,2 8.1 Hz, H-1III), 4.91 (d, 1 H, J1,2 7.8 Hz,
H-1IV), 4.90 (d, 1H, Jgem 11.0 Hz, CHHPh), 4.87 (d,
1H, Jgem 10.0 Hz, CHHPh), 4.74 (d, 1H, Jgem 11.0 Hz,
CHHPh), 4.69 (d, 1H, Jgem 10.0 Hz, CHHPh), 4.66 (d,
1H, Jgem 11.0 Hz, CHHPh), 4.58 (t, 1H, J3,4 2.8 Hz,
H-3III), 4.35 (d, 1H, J1,2 8.1 Hz, H-1I), 4.30 (d, 1H,
J1,2 8.1 Hz, H-1II), 4.05 (d, 1H, J3,4 2.8 Hz, H-4III),
3.98 (m, 1H, CHHCH2Si), 3.86 (d, 1H, J3,4 2.8 Hz, H-
4II), 3.84 (t, 1H, J3,4 9.3 Hz, J4,5 9.4 Hz, H-4I), 3.71
(dd, 1H, J5,6a 4.2 Hz, J6a,6b 10.8 Hz, H-6aI), 3.66 (m,
1H, J5,6b 10.6 Hz, J6a,6b 10.8 Hz, H-6bI), 3.57 (m, 1H,
CHHCH2Si), 3.50 (t, 1H, J3,4 9.3 Hz, H-3I), 3.40 (H-
2III), 3.38 (H-2I), 3.38 (H-2II), 3.33 (m, 1H, J4,5
9.4 Hz, J5,6a 4.2 Hz, J5,6b 10.6 Hz, H-5I), 3.19 (dd, 1H,
J3,4 2.8 Hz, H-3II), 1.00 (m, 5H, COCH3, CH2CH2Si),
0.00 (s, 9H, 3SiCH3). 13C NMR (CDCl3): d 165.95,
165.52, 165.40, 164.79, 138.81, 138.61, 138.45, 138.38,
138.16, 138.06, 133.66, 133.51, 133.31, 133.26, 129.98,
129.75, 129.66, 129.22, 129.07, 128.77, 128.64, 128.54,
128.43, 128.34, 128.32, 128.25, 128.23, 128.19, 128.15,
128.10, 127.96, 127.78, 127.65, 127.62, 127.56, 127.54,
127.51, 127.49, 127.47, 127.36, 103.13 (C-1I), 102.54
(C-1II), 102.23 (C-1III), 100.01 (C-1IV), 82.96 (C-3I),
81.88, 81.14 (C-3IV), 80.32, 79.82, 76.73 (C-4I), 75.69,
75.18, 75.12, 74.93, 74.00, 73.35, 73.13, 73.05, 72.34,
71.80, 71.44, 71.40, 69.37, 68.32, 68.25, 68.08, 67.66,
67.28, 62.42, 62.32, 54.06 (C-2III), 22.64 (COCH3),
To a solution of 6 (543 mg, 0.42 mmol) and 7 (466 mg,
0.60 mmol) in anhyd CH2Cl2 (11 mL) were added
˚
molecular sieves 4 A AW300 (500 mg) and NIS
(242 mg, 1.08 mmol), then cooled to 0 °C. To the mix-
ture was added TfOH (5.3 lL, 0.06 mmol), and the stir-
ring was continued for 30 min at 0 °C. The reaction was
quenched with Et3N (50.0 lL). The precipitate was fil-
tered off and washed with CH2Cl2. The filtrate and
washings were combined, and the solution was succes-
sively washed with satd aq NaHCO3, M Na2S2O3 and
satd aq NaCl, dried and concentrated. Purification by
flash column chromatography (1:4 EtOAc–hexane) gave
8 (453 mg, 58%) as an amorphous mass. [a]D +61.8 (c
1
1.00, CHCl3); H NMR (CDCl3): d 8.08–6.79 (m, 54H,
10Ph, p-MeOC6H4), 5.95 (d, 1H, J3,4 3.2 Hz, H-4IV),
5.77 (dd, 1H, J1,2 7.9 Hz, J2,3 10.4 Hz, H-2IV), 5.56
(dd, 1H, J2,3 10.4 Hz, J3,4 3.2 Hz, H-3IV), 5.14 (d, 1H,
J2,NH 6.4 Hz, NH), 5.07 (d, 1H, J1,2 8.3 Hz, H-1III),
4.92 (d, 1H, J1,2 7.9 Hz, H-1IV), 4.89–4.18 (14d, 14H,
12CHHPh, p-MeOC6H4CH2), 4.52 (m, 1H, J3,4 3.0 Hz,
H-3III), 4.37 (d, 1H, J1,2 8.3 Hz, H-1II), 4.35 (d, 1H,
J1,2 8.1 Hz, H-1I), 4.16 (d, 1H, J3,4 3.0 Hz, H-4III),
4.00 (d, 1H, J3,4 2.6 Hz, H-4II), 3.97 (m, 1H,
CHHCH2Si), 3.86 (t, 1H, J3,4 9.3 Hz, H-4I), 3.71 (s,
3H, OCH3), 3.57 (m, 1H, CHHCH2Si), 3.50 (t, 1H,
J3,4 9.3 Hz, H-3I), 3.38 (H-2II), 3.37 (H-2III), 3.34 (H-
5I), 3.33 (H-2I), 3.28 (H-3II), 1.04 (s, 3H, COCH3),
1.01 (m, 2H, CH2CH2Si), 0.00 (s, 9H, 3SiCH3). 13C
NMR (CDCl3): d 165.94, 165.57, 165.47, 159.20,
138.85, 138.79, 138.55, 138.35, 138.08, 133.65, 133.33,
133.28, 130.19, 130.02, 129.78, 129.72, 129.30, 129.24,
128.90, 128.67, 128.55, 128.50, 128.42, 128.35, 128.26,
128.21, 128.14, 128.10, 127.99, 127.92, 127.74, 127.60,
127.53, 127.48, 127.37, 127.25, 113.80, 103.13 (C-1I),

Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
217
18.44 (CH2CH2Si), ꢀ1.45 (SiCH3). Anal. Calcd for
101H109NO25Si: C, 68.73; H, 6.22; N, 0.79. Found:
C, 68.59; H, 6.48; N, 0.67.
76.35, 75.47, 75.20, 75.14, 74.88, 73.85, 73.44, 73.24,
73.07, 73.02, 72.31, 72.07, 72.00, 71.79, 71.42, 71.28,
69.60, 69.01, 68.53, 68.44, 68.37, 67.91, 67.79, 67.52,
67.43, 67.22, 63.06, 62.00 (C-9V), 61.60, 54.25 (C-2III),
52.93 (OCH3), 51.67, 37.65, 29.62, 29.51, 22.81, 20.94,
20.65, 20.58, 18.43, ꢀ1.45 (SiCH3). Anal. Calcd for
C
3.9. 2-(Trimethylsilyl)ethyl O-(2,3,4,6-tetra-O-benzoyl-b-
D-galactopyranosyl)-(1!3)-[O-(methyl 4,7,8,9-tetra-O-
acetyl-3,5-dideoxy-5-trichloroacetamido-D-glycero-a-D-
galacto-2-nonulopyranosylonate)-(2!6)]-O-(2-acetam-
ido-2-deoxy-b-D-galactopyranosyl)-(1!4)-O-(2,3,6-tri-
O-benzyl-b-D-galactopyranosyl)-(1!4)-2,3,6-tri-O-
benzyl-b-D-glucopyranoside (11)
C
121H133Cl3N2O37Si: C, 62.06; H, 5.72; N, 1.20. Found:
C, 5.49; H, 5.89; N, 1.06.
3.10. 2-(Trimethylsilyl)ethyl O-(2,3,4,6-tetra-O-benzoyl-
b-D-galactopyranosyl)-(1!3)-[O-(methyl 5-acetamido-
4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero-a-D-
galacto-2-nonulopyranosylonate)-(2!6)]-O-(2-acetam-
ido-2-deoxy-b-D-galactopyranosyl)-(1!4)-O-(2,3,6-tri-
O-benzyl-b-D-galactopyranosyl)-(1!4)-2,3,6-tri-O-benz-
yl-b-D-glucopyranoside (12)
To a solution of 9 (253 mg, 0.14 mmol) and ethyl
(methyl 4,7,8,9-tetra-O-acetyl-3,5-dideoxy-1-thio-5-tri-
chloroacetamido-D-glycero-D-galacto-2-nonulopyrano-
sid)onate11 (10, 269 mg, 0.42 mmol) in anhyd CH3CN
˚
(3.5 mL) were added molecular sieves 3 A AW300
(380 mg) and NIS (190 mg, 0.84 mmol), then cooled to
ꢀ40 °C. To the mixture was added TfOH (9.3 lL,
0.11 mmol), and stirring was continued for 1.5 h at
ꢀ40 °C. The reaction was quenched with Et3N
(100 lL). The precipitate was filtered off and washed
with CH2Cl2. The filtrate and washings were combined,
and the solution was successively washed with satd aq
NaHCO3, M Na2S2O3 and satd aq NaCl, dried and con-
centrated. Purification by flash column chromatography
(1:3 EtOAc–hexane) gave 11 (200 mg, 60%) as an amor-
phous mass. [a]D +36.8 (c 1.00, CHCl3); 1H NMR
(CDCl3): d 8.08–7.15 (m, 50H, 10Ph), 6.63 (d, 1H,
NHV), 5.97 (d, 1H, J3,4 3.1 Hz, H-4IV), 5.76 (dd, 1H,
J1,2 8.5 Hz, J2,3 10.0 Hz, H-2IV), 5.59 (dd, 1H, J2,3
10.0 Hz, J3,4 3.1 Hz, H-3IV), 5.32 (H-8V), 5.29 (H-7V),
5.17 (d, 1H, JNH,2 6.4 Hz, NHIII), 5.13 (d, 1H, J1,2
8.5 Hz, H-1III), 5.06 (ddd, 1H, J3ax,4 12.2 Hz, J4,5
4.5 Hz, H-4V), 4.95 (d, 1H, J1,2 8.5 Hz, H-1IV), 4.88 (d,
1H, Jgem 11.3 Hz, CHHPh), 4.85 (d, 1H, Jgem 11.3 Hz,
CHHPh), 4.56 (H-3III), 4.47 (d, 1H, Jgem 12.1 Hz,
CHHPh), 4.38 (d, J1,2 7.6 Hz, H-1II), 4.35 (d, J1,2
8.7 Hz, H-1I), 4.29 (H-6V), 4.20 (H-4III), 4.19 (H-9aV),
4.00 (H-9bV), 3.98 (m, 1H, CHHCH2Si), 3.88 (J4,5
4.5 Hz, H-5V), 3.87 (J4,5 9.1 Hz, H-4I), 3.77 (s, 3H,
COOCH3), 3.58 (m, 1H, CHHCH2Si), 3.50 (t, 1H, J3,4
9.1 Hz, H-3I), 3.38 (H-2II), 3.36 (H-5I), 3.34 (H-3II),
3.32 (H-2III), 3.30 (H-2I), 2.60 (m, 1H, Jgem 12.2 Hz,
H-3eqV), 2.10, 2.09, 1.95, 1.91, 1.10 (s, each 3H,
5COCH3), 1.83 (t, 1H, Jgem = J3ax,4 = 12.2 Hz, H-3axV),
1.00 (m, 2H, CH2CH2Si), 0.00 (s, 9H, 3SiCH3). 13C
NMR (CDCl3): d 177.60, 170.62, 170.49, 170.31,
169.88, 169.69, 167.81, 165.84, 165.49, 165.44, 164.84,
161.95, 138.92, 138.84, 138.78, 138.54, 138.33, 138.12,
133.59, 133.33, 133.25, 129.96, 129.76, 129.66, 129.25,
129.21, 128.97, 128.62, 128.50, 128.44, 128.36, 128.21,
128.17, 128.11, 128.06, 128.00, 127.93, 127.81, 127.72,
127.64, 127.57, 127.44, 127.30, 127.20, 103.09, (C-1I),
102.37 (C-1II), 102.33 (C-1IV), 99.24 (C-1III), 98.92,
92.06, 82.98 (C-3I), 81.91, 81.87, 79.82, 79.62 (C-3III),
A solution of 11 (193 mg, 82.5 lmol), tributyltin hydride
(197 lL, 0.74 mmol), N,N-dimethylacetamide (0.54 mL)
and 2,20-azobisisobutyronitrile (AIBN; 3.3 mg) in anhyd
benzene (3.3 mL) heated under reflux for 15 min. The
reaction mixture was then cooled and concentrated
under diminished pressure. Purification by flash column
chromatography (2:1 EtOAc–hexane) gave 12 (178 mg,
96%) as an amorphous mass. [a]D +38.5 (c 1.00, CHCl3);
1H NMR (CDCl3): d 8.15–7.10 (m, 50H, 10Ph), 5.97
(d,1H, J3,4 3.4 Hz, H-4IV), 5.76 (dd, 1H, J1,2 7.9 Hz,
J2,3 10.2 Hz, H-2IV), 5.58 (dd, 1H, J2,3 10.2 Hz, J3,4
3.4 Hz, H-3IV), 5.34–5.28 (m, 3H, H-7V, H-8V, NH),
5.11 (d, 1H, J1,2 8.3 Hz, H-1III), 4.96 (d, 1H, J1,2
7.9 Hz, H-1IV), 4.82 (m, 1H, H-4V), 4.52 (m, 1H, H-
3III), 4.38 (d, 1 H, J1,2 7.6 Hz, H-1II), 4.35 (d, 1H, J1,2
8.3 Hz, H-1I), 4.21–4.17 (m, 2H, H-4III, H-9aV), 4.12
(d, 1H, J3,4 2.2 Hz, H-4II), 3.99–3.93 (m, 2H, H-9bV,
CHHCH2Si), 3.86 (t, 1H, J3,4 9.1 Hz, H-4I), 3.75 (s,
3H, COOCH3), 3.69–3.67 (m, 3H, H-6aI, H-6bI, H-
6aII), 3.56 (m, 1H, CHHCH2Si), 3.49 (t, 1H, J3,4
9.1 Hz, H-3I), 3.43 (m, 1H, H-5II), 3.40–3.27 (m, 6H,
H-2II, H-2III, H-5I, H-3II, H-2I, H-6bII), 2.54 (dd, 1H,
Jgem 12.8 Hz, J3eq,4 4.5 Hz, H-3eqV), 1.85 (m, 1H, Jgem
12.8 Hz, H-3axV), 2.08, 2.07, 1.95, 1.92, 1.84, 1.10 (6s,
each 3H, 6COCH3), 1.00 (m, 2H, CH2CH2Si), 0.00 (s,
9H, 3SiCH3). 13C NMR (CDCl3): d 170.75, 170.65,
170.54, 170.21, 169.96, 169.60, 168.00, 165.83, 165.50,
165.41, 164.82, 38.97, 138.86, 138.77, 138.58, 138.35,
138.13, 133.58, 133.33, 133.23, 133.20, 129.95, 129.78,
129.75, 129.68, 129.26, 128.99, 128.62, 128.58, 128.51,
128.44, 128.36, 128.31, 128.22, 128.19, 128.18, 128.11,
128.08, 128.06, 128.01, 127.95, 127.89, 127.85, 127.73,
127.71, 127.69, 127.62, 127.57, 127.48, 127.44, 127.42,
127.31, 127.18, 103.10 (C-1I), 102.34 (C-1II, C-1IV),
98.95 (C-1III), 82.98 (C-3I), 81.93, 81.90, 79.84, 79.72
(C-3III), 76.38, 75.49, 75.22, 75.15, 74.89, 73.95, 73.28,
73.08, 72.64, 72.30, 72.05, 71.43, 71.30, 69.63, 69.07,
69.02, 68.59, 68.39, 67.91, 67.45, 67.38, 67.23
(CH2CH2Si), 62.91, 62.17 (C-9V), 61.60, 54.23 (C-2III),

218
Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
52.75 (OCH3), 49.45 (C-5V), 37.59 (C-3V), 23.11, 22.80,
20.96, 20.76, 20.74, 20.65, 18.44 (CH2CH2Si), ꢀ1.45
(SiCH3). Anal. Calcd for C121H136N2O37Si: C, 62.92;
H, 6.12; N, 1.25. Found: C, 62.83; H, 6.26; N, 1.25.
1III), 96.38 (C-1V), 74.28, 74.10 (C-4I), 73.59 (C-3II),
73.16 (C-3I), 73.08, 72.56, 71.81 (C-2I), 71.76 (C-3IV),
71.63, 70.72, 70.30 (C-2II), 70.07 (C-2IV), 69.73, 69.01,
68.99, 68.77 (C-4II), 67.81, 67.49, 67.38 (CH2CH2Si),
63.65, 62.56, 62.00, 61.89, 61.83 (OCH3), 56.52, 55.04,
53.38, 52.39, 49.60, 37.31, 29.65, 23.11, 20.78, 20.74,
20.69, 20.65, 17.84 (CH2CH2Si), ꢀ1.58 (SiCH3). Anal.
Calcd for C128H128N2O44Si: C, 63.36; H, 5.32; N, 1.15.
Found: C, 63.24; H, 5.60; N, 1.02.
3.11. 2-(Trimethylsilyl)ethyl O-(2,3,4,6-tetra-O-benzoyl-
b-D-galactopyranosyl)-(1!3)-[O-(methyl 5-acetamido-
4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero-a-D-
galacto-2-nonulopyranosylonate)-(2!6)]-O-(2-acetam-
ido-4-O-benzoyl-2-deoxy-b-D-galactopyranosyl)-(1!4)-
O-(2,3,6-tri-O-benzoyl-b-D-galactopyranosyl)-(1!4)-
2,3,6-tri-O-benzoyl-b-D-glucopyranoside (13)
3.12. O-(2,3,4,6-Tetra-O-benzoyl-b-D-galactopyranosyl)-
(1!3)-[O-(methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-
3,5-dideoxy-D-glycero-a-D-galacto-2-nonulopyranosylo-
nate)-(2!6)]-O-(2-acetamido-4-O-benzoyl-2-deoxy-b-D-
galactopyranosyl)-(1!4)-O-(2,3,6-tri-O-benzoyl-b-D-
galactopyranosyl)-(1!4)-O-(2,3,6-tri-O-benzoyl-b-D-
glucopyranosyl)-(1!1)-(2S,3R,4E)-2-azido-3-O-benzoyl-
4-eicosene-1,3-diol (15)
Compound 12 (136 mg, 60.8 lmol) was dissolved in
EtOAc (5.4 mL) and then a solution of sodium bromate
(165 mg, 1.1 mmol) in water (3.7 mL) was added. To the
well stirred two-phase system an aqueous solution of so-
dium hydrosulfite (80% pure, 203 mg, dissolved in
7.7 mL water) was added dropwise over 20 min at rt.
The mixture was diluted with EtOAc and the organic
phase was successively washed with M Na2S2O3 and
satd aq NaCl, dried (MgSO4) and concentrated under
diminished pressure. To the solution of the residue in
pyridine (2.5 mL) added dropwise benzoyl chloride for
2.5 h at rt. The reaction was quenched with MeOH
(60 lL) and diluted with CH2Cl2, successively washed
with satd aq NaHCO3 and satd aq NaCl, dried and con-
centrated. Purification by flash column chromatography
(1:80 MeOH–CH2Cl2) gave 13 (80 mg, 59%) as an amor-
phous mass. [a]D +21.1 (c 1.00, CHCl3); 1H NMR
(CDCl3): d 8.31–7.02 (m, 55H, 11Ph), 6.07 (d, 1H, J3,4
3.4 Hz, H-4IV), 5.84 (dd, 1H, J1,2 7.7 Hz, J2,3 10.2 Hz,
H-2IV), 5.74 (t, 1H, J2,3 9.6 Hz, J3,4 9.8 Hz, H-3I), 5.69
(dd, 1H, J2,3 10.2 Hz, J3,4 3.4 Hz, H-3IV), 5.59 (dd,
1H, J1,2 7.8 Hz, J2,3 10.5 Hz, H-2II), 5.51 (dd, 1H, J2,3
9.6 Hz, J1,2 7.9 Hz, H-2I), 5.40 (dd, 1H, J2,3 10.5 Hz,
J3,4 2.3 Hz, H-3II), 5.35 (d, 1H, J1,2 7.8 Hz, H-1III),
5.25 (d, 1H, NH), 5.20 (d, 1H, J1,2 7.7 Hz, H-1IV),
4.91 (d, 1H, J1,2 7.8 Hz, H-1II), 4.87 (m, 1H, J3eq,4
4.7 Hz, H-4V), 4.74 (d, 1H, J1,2 7.9 Hz, H-1I), 4.59 (d,
1H, J3,4 2.3 Hz, H-4II), 4.35 (t, 1H, J3,4 9.8 Hz, H-4I),
4.33 (t, 1H, J1,2 7.8 Hz, J2,3 12.8 Hz, H-2III), 4.09 (dd,
1H, J2,3 12.8 Hz, H-3III), 4.03–3.97 (m, 2H, H-5V,
CHHCH2Si), 3.88 (m, 1H, H-5I), 3.43 (s, 3H,
COOCH3), 2.48 (dd, 1H, Jgem 13.0 Hz, J3eq,4 4.5 Hz,
H-3eqV), 1.78 (m, 1H, H-3axV), 2.16, 2.08, 2.07, 20.6,
2.01, 1.94 (6s, each 3H, 6COCH3), 0.95 (m, 2H,
CH2CH2Si), 0.00 (s, 9H, 3SiCH3). 13C NMR (CDCl3):
d 170.87, 170.42, 170.09, 169.96, 169.61, 167.48,
165.90, 165.84, 165.56, 165.46, 165.43, 164.96, 164.86,
133.54, 133.48, 133.27, 133.23, 133.11, 132.62, 130.99,
130.26, 130.15, 130.12, 130.02, 129.94, 129.91, 129.79,
129.75, 129.69, 129.63, 129.53, 129.50, 129.43, 129.29,
129.25, 129.17, 128.96, 128.95, 128.89, 128.72, 128.68,
128.61, 128.55, 128.51, 128.43, 128.35, 128.23, 128.18,
101.53 (C-1IV), 100.54 (C-1I), 99.42 (C-1II), 98.41 (C-
To a solution of 13 (131 mg, 54.0 lmol) in anhyd
CH2Cl2 (2.0 mL) was added TFA (2.5 mL) at 0 °C and
the mixture was stirred for 30 min at 0 °C, quenched
with propylacetate (5.0 mL) and concentrated. The res-
idue was then dissolved in anhyd CH2Cl2 (2.0 mL) and
CCl3CN (126 lL, 1.25 mmol) and 1,8-diazabicy-
clo[5.4.0]undec-7-ene (DBU; 13 lL, 87.0 lmol) were
added at rt. The mixture was stirred for 1 h at rt, then
concentrated. The residue was filtered through short
pad of silica gel and concentrated. To a solution of
the residue and (2S,3R,4E)-2-azido-3-O-benzoyl-4-eico-
sene-1,3-diol12 (14, 40 mg, 87.4 lmol) in anhyd CH2Cl2
˚
were added molecular sieves 4 A AW300 (130 mg) and
BF3ÆEt2O (8.5 lL, 67.0 lmol). The mixture was stirred
for 2 h at 0 °C and diluted with anhyd CH2Cl2 and suc-
cessively washed with satd aq NaHCO3 and satd aq
NaCl, dried and concentrated. Purification by flash col-
umn chromatography (1:80 MeOH–CH2Cl2) gave 15
(54 mg, 36%) as an amorphous mass. [a]D +14.4 (c
1
1.00, CHCl3); H NMR (CDCl3): d 8.04–6.93 (m, 60H,
12Ph), 5.87 (d, 1H, J3,4 3.8 Hz, H-4IV), 5.64 (dd, 1H,
J1,2 7.7 Hz, J2,3 10.4 Hz, H-2IV), 5.60 (m, 1H, J4,5
0
15.9 Hz, J5,6 = J5,6 6.8 Hz, H-5 of sphingosine), 5.56
(t, 1H, J2,3 9.4 Hz, J3,4 9.4 Hz, H-3I), 5.49 (dd, 1H,
J2,3 10.4 Hz, J3,4 3.8 Hz, H-3IV), 5.43 (t, 1H, J3,4
7.9 Hz, H-3 of sphingosine), 5.39 (dd, 1H, J1,2 7.6 Hz,
J2,3 10.6 Hz, H-2II), 5.34 (m, 1H, J3,4 7.9 Hz, J4,5
15.9 Hz, H-4 of sphingosine), 5.33 (dd, 1H, J1,2
7.6 Hz, J2,3 9.4 Hz, H-2I), 5.21 (dd, 1H, J2,3 10.6 Hz,
J3,4 2.5 Hz, H-3II), 5.16 (d, 1H, J3,4 2.1 Hz, H-4III),
5.15 (d, 1H, J1,2 5.3 Hz, H-1III), 4.99 (d, 1H, J1,2
7.7 Hz, H-1IV), 4.98 (d, 1H, JNH,5 9.8 Hz, NHV), 4.71
(d, 1H, J1,2 7.6 Hz, H-1II), 4.64 (m, 1H, H-4V), 4.54
(d, 1H, J1,2 7.6 Hz, H-1I), 4.40 (d, 1H, J3,4 2.5 Hz, H-
4II), 4.17 (t, 1H, J3,4 9.4 Hz, H-4I), 4.13 (dd, 1H, J2,3
12.5 Hz, J3,4 2.1 Hz, H-3III), 3.89 (dd, 1H, J1,2 5.3 Hz,
J2,3 12.5 Hz, H-2III), 3.68 (m, 1H, H-5I), 3.22 (s, 3H,

Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
219
COOCH3), 2.28 (dd, 1H, Jgem 12.9 Hz, H-3eqV), 1.96,
1.88, 1.87, 1.86, 1.79, 1.74 (6s, each 3H, 6COCH3),
1.57 (t, 1H, Jgem 12.9 Hz, H-3axV), 1.40–1.04 (m, 26H,
13CH2), 0.80 (t, 3H, CH2CH3). 13C NMR (CDCl3): d
140.45 (C-5 of sphingosine), 136.25–128.86 (arom-C),
124.02 (C-4 of sphingosine), 103.12 (C-1IV), 102.55 (C-
1I), 100.00 (C-1II), 97.99 (C-1III), 76.26, 75.88 (C-4III),
75.34 (C-4I), 75.15 (C-3II), 74.96, 74.32 (C-3I), 74.14,
73.34, 73.23 (C-3IV), 73.14 (C-2I), 72.30, 71.84 (C-2IV),
71.66, 70.57, 70.36, 69.80, 69.73, 69.39 (C-4IV), 68.96,
64.99, 63.88, 63.59, 63.49, 63.47, 63.44, 63.41, 63.88
(C-3III), 63.59, 63.49, 63.47 (C-2III), 63.44, 63.41,
58.01, 54.07, 51.19, 38.80, 33.86, 24.78, 24.77, 22.27,
22.19, 15.66. The carbonyl carbons and the quaternary
carbon (C-2V) of 15 could not be assigned, because
these chemical shifts of the carbons were recorded by
1H–13C correlation spectroscopy. Anal. Calcd for
4.74 (d, 1H, J1,2 7.5 Hz, H-1II), 4.71 (m, 1H, J3eq,4
4.7 Hz, H-4V), 4.51 (d, 1H, J1,2 7.9 Hz, H-1I), 4.46 (d,
1H, J3,4 2.4 Hz, H-4II), 3.68 (m, 1H, H-5I), 3.26 (s,
3H, OCH3), 2.35 (dd, 1H, Jgem 13.2 Hz, J3eq,4 4.7 Hz,
H-3Veq), 2.03 (s, 3H, COCH3), 1.96 (m, 2H, H-6 of
sphingosine), 1.95, 1.94, 1.93, 1.86, 1.82 (6s, each 3H,
5COCH3), 1.78 (t, 1H, Jgem 13.2 Hz, H-3axV), 1.40–
1.04 (m, 54H, 27CH2 of sphingosine and octadecan-
amide), 0.88 (t, 6H, 2CH2CH3 of sphingosine and octa-
decanamide). 13C NMR (CDCl3): d 172.54, 172.23,
170.93, 170.48, 170.11, 169.95, 169.60, 167.38, 165.89,
165.79, 165.66, 165.54, 165.50, 165.44, 165.41, 165.03,
165.00, 164.95, 164.67, 164.40, 137.44 (C-5 of sphingo-
sine), 133.61, 133.58, 133.50, 133.34, 133.31, 133.24,
133.18, 133.13, 132.79, 132.66, 130.89, 130.16, 130.11,
130.08, 129.92, 129.89, 129.77, 129.74, 129.66, 129.59,
129.49, 129.41, 129.27, 129.25, 129.18, 129.09, 129.08,
128.82, 128.80, 128.70, 128.60, 128.56, 128.51, 128.46,
128.44, 128.31, 128.19, 128.18, 124.75 (C-4 of sphingo-
sine), 101.50 (C-1IV), 100.90 (C-1I), 99.14 (C-1II), 96.24
(C-1III), 74.30, 73.85, 73.46, 73.43, 73.21, 72.48, 72.42,
72.39, 72.01, 71.60, 70.58, 70.14, 69.99, 69.59, 68.80,
68.70, 68.55, 68.52, 67.69, 67.51, 67.22, 63.61, 61.92,
61.76, 61.71, 56.39, 52.38, 50.27, 49.49, 37.26, 36.43,
33.53, 32.27, 31.90, 29.68, 29.67, 29.64, 29.61, 29.51,
29.48, 29.45, 29.34, 29.25, 29.23, 29.16, 29.06, 28.91,
25.48, 24.76, 23.32, 23.15, 22.66, 20.81, 20.77, 20.70,
14.12. The carbonyl carbons and the quaternary carbon
(C-2V) of 16 could not be assigned, because these chem-
ical shifts of the carbons were recorded by 1H–13C
correlation spectroscopy. HRFABMS: calcd for
C150H157N5O46: C, 65.14; H, 5.72; N, 2.53. Found: C,
64.09; H, 5.81; N, 2.61.
3.13. O-(2,3,4,6-Tetra-O-benzoyl-b-D-galactopyranosyl)-
(1!3)-[O-(methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-
3,5-dideoxy-D-glycero-a-D-galacto-2-nonulopyranosylo-
nate)-(2!6)]-O-(2-acetamido-4-O-benzoyl-2-deoxy-b-D-
galactopyranosyl)-(1!4)-O-(2,3,6-tri-O-benzoyl-b-D-
galactopyranosyl)-(1!4)-O-(2,3,6-tri-O-benzoyl-b-D-
glucopyranosyl)-(1!1)-(2S,3R,4E)-3-O-benzoyl-4-eico-
sene-2-octadecanamido-1,3-diol (16)
A mixture of 15 (46 mg, 16.6 lmol), zinc powder
(370 mg) and acetic acid (27 lL) in anhyd CH2Cl2
(2.6 mL) was stirred at rt for 4 h and then filtered
through Celite. The filtrate was concentrated under
diminished pressure and remaining AcOH was co-evap-
orated with toluene. The residue filtered through short
pad of silica gel and concentrated. To a solution of the
residue (34 mg) and octadecanoic acid (35 mg,
0.12 mmol) in anhyd CH2Cl2 (2.5 mL) was added N-
(3-dimethylaminopropyl)-N0-ethylcarbodiimide hydro-
chloride (12 mg, 0.06 mmol) and stirred for 12 h at rt.
The reaction mixture was diluted with anhyd CH2Cl2
and was successively washed with water, dried and con-
centrated. Purification by flash column chromatography
(1:80 MeOH–CH2Cl2) gave 16 (37 mg, 74%). [a]D +24.2
(c 1.00, CHCl3); 1H NMR (CDCl3): d 8.17–6.96 (m,
60H, 12Ph), 5.95 (d, 1H, J3,4 3.8 Hz, H-4IV), 5.82 (m,
C
3005.2188.
168H193N3O47 [M+H]+: m/z 3005.2883, found
3.14. O-(b-D-Galactopyranosyl)-(1!3)-[O-(5-acetamido-
3,5-dideoxy-D-glycero-a-D-galacto-2-nonulopyranosylo-
nate)-(2!6)]-O-(2-acetamido-2-deoxy-b-D-galactopyr-
anosyl)-(1!4)-O-(b-D-galactopyranosyl)-(1!4)-O-(b-D-
glucopyranosyl)-(1!1)-(2S,3R,4E)-4-eicosene-2-octa-
decanamido-1,3-diol (17)
To a solution of 16 (15 mg, 5.0 lmol) in anhyd MeOH
(1.5 mL) was added 0.5 M NaOMe in anhyd MeOH
(70.0 lL), and the mixture was stirred for 24 h at rt,
and water (0.5 mL) was added. The solution was stirred
for 17 h at rt, then acidified with Amberlite IR-120 (H+)
resin until pH 4 (pH-indicator paper), and the mixture
was filtered. The resin was washed with MeOH, and
combined filtrate and washings were concentrated. Puri-
fication by column chromatography (7:40:50 H2O–
MeOH–CHCl3) of the residue on Sephadex LH-20
(8 g) gave 17 (6 mg, 76%). [a]D ꢀ11.0 (c 0.34, 7:40:50
0
1H, J4,5 15.2 Hz, J5,6 = J5,6 = 7.6 Hz, H-5 of sphingo-
sine), 5.71 (dd, 1H, J1,2 7.9 Hz, J2,3 10.4 Hz, H-2IV),
5.62 (t, 1H, J2,3 9.7 Hz, H-3I), 5.59 (d, 1H, JNH,2
9.4 Hz, NH of sphingosine), 5.55 (dd, 1H, J2,3 10.4 Hz,
J3,4 3.8 Hz, H-3IV), 5.47 (t, 1H, J3,4 7.7 Hz, H-3 of sphin-
gosine), 5.43 (dd, 1H, J1,2 7.5 Hz, J2,3 10.6 Hz, H-2II),
5.40 (dd, 1H, J3,4 7.7 Hz, J4,5 15.2 Hz, H-4 of sphingo-
sine), 5.34 (dd, 1H, J1,2 7.9 Hz, J2,3 9.7 Hz, H-2I), 5.27
(dd, 1H, J2,3 10.6 Hz, J3,4 2.4 Hz, H-3II), 5.09 (d, 1H,
JNH,5 9.2 Hz, NHV), 5.06 (d, 1H, J1,2 7.9 Hz, H-1IV),
1
H2O–MeOH–CHCl3); H NMR (1:3:3 D2O–CD3OD–
CDCl3): d 5.70 (m, 1H, J4,5 14.8 Hz, H-5 of sphingo-
sine), 5.44 (dd, 1H, J3,4 8.0 Hz, J4,5 14.8 Hz, H-4 of
sphingosine), 4.63 (d, 1H, J1,2 8.5 Hz, H-1IV), 4.37 (d,

220
Y. Takeda, S. Horito / Carbohydrate Research 340 (2005) 211–220
1H, J1,2 7.7 Hz, H-1II), 4.35 (d, 1H, J1,2 7.6 Hz, H-1III),
4.31 (d, 1H, J1,2 7.7 Hz, H-1I), 4.12 (d, 1H, J3,4 2.8 Hz,
H-4IV), 4.08 (t, 1H, J3,4 8.0 Hz, H-3 of sphingosine),
4.04 (d, 1H, J3,4 2.5 Hz, H-4II), 4.00 (dd, 1H, J1,2
8.5 Hz, J2,3 10.8 Hz, H-2IV), 3.83 (d, 1H, J3,4 3.0 Hz,
H-4III), 3.75 (dd, 1H, J2,3 10.8 Hz, J3,4 2.8 Hz, H-3IV),
3.64 (dd, 1H, J2,3 10.1 Hz, J3,4 2.5 Hz, H-3II), 3.54 (dd,
1H, J2,3 9.7 Hz, J1,2 7.6 Hz, H-2III), 3.49 (dd, 1H, J2,3
9.7 Hz, J3,4 3.0 Hz, H-3III), 3.43 (dd, 1H, J1,2 7.7 Hz,
J2,3 10.1 Hz, H-2II), 2.77 (dd, 1H, Jgem 12.0 Hz, J3eq,4
4.2 Hz, H-3eqV), 2.18 (t, 2H, COCH2), 2.04, 2.02 (2 s,
each 3H, 2NCOCH3), 2.03 (m, 2H, H-6, 60 of sphingo-
sine), 1.61 (t, 1H, Jgem 12.0 Hz, H-3axV), 1.58 (m, 2H,
COCH2CH2), 1.33–1.23 (m, 54H, 27CH2 of sphingosine
and octadecanamide), 0.89 (t, 6H, 2CH2CH3 of sphingo-
sine and octadecanamide). Signals overlapping with the
signal of solvents could not be assigned. 13C NMR (1:3:3
D2O–CD3OD–CDCl3): d 135.37 (C-5 of sphingosine),
129.96 (C-4 of sphingosine), 105.58 (C-1III), 103.92 (C-
1II), 103.49 (C-1I), 103.18 (C-1IV), 80.90, 79.74, 76.80,
75.69, 75.58, 73.80, 73.75, 73.58, 73.53, 72.53, 72.19,
71.75, 71.58, 70.75, 69.64, 69.36, 68.87, 68.81, 64.32,
64.21, 63.87, 63.82, 61.60, 61.54, 61.26, 60.88, 53.66,
52.83, 52.22, 41.36, 41.06, 36.97, 33.05, 30.07, 26.58,
22.78, 23.14, 18.93, 14.27. The carbonyl carbons and
the quaternary carbon (C-2V) of 17 could not be as-
signed, because these chemical shifts of the carbons
were recorded by 1H–13C correlation spectroscopy.
HRFABMS: calcd for C168H193N3O47 [M+H]+: m/z
1573.9080, found 1573.9091.
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