Synthesis of a mannose heptasaccharide of the pathogenic yeast, Candida glabrata IFO 0622 strain

Article abstract of DOI:10.1016/S0008-6215(02)00162-3

An effective synthesis of the mannose heptasaccharide existing in the pathogenic yeast, Candida glabrata IFO 0622 strain was achieved via TMSOTf-promoted condensation of a tetrasaccharide donor 13 with a trisaccharide acceptor 16, followed by deprotection

Full text of DOI:10.1016/S0008-6215(02)00162-3

Carbohydrate Research 337 (2002) 1367–1371
www.elsevier.com/locate/carres
Synthesis of a mannose heptasaccharide of the pathogenic yeast,
Candida glabrata IFO 0622 strain
Youlin Zeng, Jianjun Zhang, Fanzuo Kong*
Research Center for Eco-En6ironmental Sciences, Academia Sinica, Chinese Academy of Sciences, PO Box 2871, Beijing 100085, PR China
Received 12 April 2002; accepted 9 May 2002
Abstract
An effective synthesis of the mannose heptasaccharide existing in the pathogenic yeast, Candida glabrata IFO 0622 strain was
achieved via TMSOTf-promoted condensation of a tetrasaccharide donor 13 with a trisaccharide acceptor 16, followed by
deprotection. The tetrasaccharide 13 was constructed by coupling of 2,3,4,6-tetra-O-benzoyl-a-
tri-O-acetyl-a- -mannopyranosyl trichloroacetimidate (7) with allyl 3,4,6-tri-O-benzoyl-a- -mannopyranosyl-(1“2)-3,4,6-tri-O-
benzoyl-a- -mannopyranoside (10), followed by deallylation and trichloroacetimadation. The trisaccharide 16 was obtained by
coupling of 6-O-acetyl-2,3,4-tri-O-benzoyl-a- -mannopyranosyl trichloroacetimidate with 10, and subsequent 6-O-deacetylation.
The disaccharide 7 was prepared through coupling of perbenzoylated mannosyl trichloroacetimidate with 4,6-O-benzylidene-1,2-
O-ethylidene-b- -mannopyranose, then simultaneous debenzylidenation and deethylidenation, and subsequent acetylation, selec-
tive 1-O-deacetylation, and trichloroacetimidation. The disaccharide 10 was obtained by self-condensation of
D-mannopyranosyl-(1“3)-2,4,6-
D
D
D
D
D
3,4,6-tri-O-benzoyl-1,2-O-allyloxyethylidene-b-D-mannopyranose, followed by selective 2-O-deacetylation. © 2002 Elsevier Sci-
ence Ltd. All rights reserved.
Keywords: Mannose oligosaccharides; Trichloroacetimidates; Regio- and stereoselective synthesis
1. Introduction
D
-Manp-(1“2)-D-Man. To the best of our knowledge,
there have been no reports dealing with the synthesis of
the heptasaccharide. For an investigation of structure–
function relationships of mannan, we present herein a
facile and convergent synthesis of the mannose hep-
tasaccharide.
Candida species are opportunistic pathogens of hu-
mans that frequently cause severe systemic infections in
patients with AIDS,1 cancer,2 and burns3 as well as in
those under immunosuppressive or radiation therapy.4
Candida glabrata as a medically important fungus has
been steadily attracting attention from microbiologists
interested in infectious disease research.5 A structural
analysis of the cell wall mannan isolated from C.
glabrata IFO 0622 strain was carried out by Suzuki’s
group.6 Three novel oligosaccharides, i.e., a tetraose, a
hexaose, and a heptaose, were obtained from mild
acetolysis of acid- and alkali-stable mannan moiety.
The tetraose and hexaose were known a-(1“2)- and
a-(1“3)-linked structures, while the heptaose was iden-
2. Results and discussion
Retrosynthetic analysis indicated that the mannose
heptamer can be obtained with a (1“6) linkage by
condensation of two moieties, i.e., a mannose tetramer
donor and a mannose trimer acceptor. The tetra-
asaccharide then can be constructed from a (1“3)-
linked disaccharide donor and
a
(1“2)-linked
tified
as
a-
D
-Manp-(1“3)-a-
D
-Manp-(1“2)-a-
D-
disaccharide acceptor, while the trisaccharide can be
built from the same (1“2)-linked disaccharide acceptor
and a mannose donor.
Manp-(1“2)-a-
D
-Manp-(1“6)-a-
D
-Manp-(1“2)-a-
Our synthetic route is shown in Scheme 1. Coupling
* Corresponding author. Tel.: +86-10-62936613; fax: +
86-10-62923563
of perbenzoylated mannosyl trichloroacetimidate7
1
E-mail address: fzkong@mail.rcees.ac.cn (F. Kong).
with 4,6-O-benzylidene-1,2-O-ethylidene-b-D-mannopy-
0008-6215/02/$ - see front matter © 2002 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(02)00162-3

1368
Y. Zeng et al. / Carbohydrate Research 337 (2002) 1367–1371
Scheme 1. Reagents and conditions: a. TMSOTf, CH2Cl2, N2, −15 °C to rt, 4 h; b. CH2Cl2, CH3OH–CH3COCl, rt; c. Ac2O/py
(dry), rt, 6 h; d. CH2Cl2, NH3–MeOH, rt; e. CCl3CN, DBU, CH2Cl2, rt, 8 h; f. PdCl2, CH2Cl2, rt, 2 h; g. NH3–MeOH, rt.
ranose (2) gave 2,3,4,6-tetra-O-benzoyl-a-
ranosyl-(1“3)-4,6-O-benzylidene-1,2-O-ethylidene-b-
-mannopyranose (3) in high yield (83%). Removal of
D
-mannopy-
deacetylation produced the trisaccharide acceptor 16.
Finally, condensation of the tetrasaccharide donor 13
with 16 gave the heptasaccharide 17, and subsequent
deacylation in ammonia-saturated methanol yielded the
target mannose heptasaccharide 18, whose bioassay is
in progress.
D
the benzylidene and ethylidene groups was readily
achieved simultaneously with 0.3:50:10 acetyl chloride–
methanol–dichloromethane within 1 h, giving the dis-
accharide 4 as a white solid in high yield (90%) after
purification. This reaction was smooth and easily con-
trolled. Acetylation of 4 with acetic anhydride in pyri-
dine, followed by selective 1-O-deacetylation with M
solution of ammonia in 1:1 methanol–dichlor-
omethane, and then trichloroacetimidation with
trichloroacetonitrile in the presence of DBU furnished
Compared to the previously reported syntheses10,12 of
complex mannans containing (1“2), (1“3), and (1“
6) linkages, the method presented herein is simpler and
convergent, owing to the sole use of acyl protection
groups. It should be possible to carry out large-scale
synthesis employing the method as described.
2,3,4,6-tetra-O-benzoyl-a-
D
-mannopyranosyl-(1“3)-
2,4,6-tri-O-acetyl-a- -mannopyranosyl trichloroacetim-
D
3. Experimental
idate (7) in 61% yield (for three steps). The disaccharide
10 was prepared by TMSOTf-promoted self condensa-
tion of 3,4,6-tri-O-benzoyl-1,2-O-allyloxyethylidene-b-
General methods.—Optical rotations were deter-
mined at 25 °C with a Perkin–Elmer model 241-Mc
automatic polarimeter. 1H, 13C NMR and 1H–13C
COSY spectra were recorded with Bruker ARX 400
D
-mannopyranose,8 followed by selective 2-O-
deacetylation with 0.5:100:50 acetyl chloride–
methanol–dichloromethane.8,9 Condensation of the dis-
accharide donor 7 with the disaccharide acceptor 10
gave the tetrasaccharide 11 in 61% yield. Deallylation10
of 11 with PdCl2 in dichloromethane furnished the
tetrasaccharide hemiacetal 12, and subsequent
trichloroacetimidation produced the tetrasaccharide
donor 13. Coupling of 6-O-acetyl-2,3,4-tri-O-benzoyl-
1
spectrometers (400 MHz for H, 100 MHz for 13C) at
25 °C for solutions in CDCl3 or D2O as indicated. Mass
spectra were measured with MALDITOF-MS spec-
trometer or recorded with a VG PLATFORM mass
spectrometer using the ESI mode. Thin-layer chro-
matography (TLC) was performed on Silica Gel HF254
with detection by charring with 30% (v/v) H2SO4 in
MeOH or in some cases by a UV lamp. Column
chromatography was conducted by elution of a column
a-
D
-mannopyranosyl trichloroacetimidate11 (14) with
the disaccharide acceptor 10, followed by selective 6-O-

Y. Zeng et al. / Carbohydrate Research 337 (2002) 1367–1371
1369
(16×240 mm, 18×300 mm, 35×400 mm) of silica gel
(100–200 mesh) with EtOAc–petroleum ether (60–
90 °C) as the eluent. Solutions were concentrated at
B60 °C under reduced pressure. The pure R-isomer13
of 1,2-O-ethylidene-b-D-mannopyranose was used for
the synthesis of 2, giving predominantly the R-isomer
of 2, and subsequently 3 as well.
azabicyclo[5.4.0]undecene (DBU) (0.50 mL, 4.04 mmol)
in dry CH2Cl2 (50 mL) was stirred under nitrogen for 3
h and then concentrated. The residue was purified by
flash chromatography (2:1 petroleum ether–EtOAc) to
give 7 (5.55 g, 61% for three steps from 5 to 7) as a
1
white foam: [h]D −8.5° (c 1.0, CHCl3); H NMR (400
MHz, CDCl3): l 8.79 (s, 1 H, NH), 8.15–7.21 (m, 20
H, 4 BzH), 6.35 (d, 1 H, J1,2 1.7 Hz, H-1), 6.20 (dd, 1
H, J3,4=J4,5 9.9 Hz, H-4%), 5.78 (dd, 1 H, J2,3 3.2, J3,4
9.9 Hz, H-3%), 5.59 (dd, 1 H, J1,2 1.4, J2,3 3.2 Hz, H-2%),
5.54 (dd, 1 H, J3,4=J4,5 10.0 Hz, H-4), 5.48 (dd, 1 H,
J1,2 1.7, J2,3 3.2 Hz, H-2), 5.37 (d, 1 H, J1,2 1.4 Hz,
H-1%), 4.64–4.56 (m, 2 H, 2 H-6%), 4.47 (dd, 1 H, J5,6
2.6, J6,6 12.1 Hz, H-6), 4.42 (dd, 1 H, J2,3 3.2, J3,4 10.0
Hz, H-3), 4.27 (dd, 1 H, J5,6 5.2, J6,6 12.4 Hz, H-6),
4.21–4.10 (m, 2 H, H-5, H-5%), 2.39 (s, 3 H, MeCO),
2.24 (s, 3 H, MeCO), 2.10 (s, 3 H, MeCO). Anal. Calcd
for C48H44Cl3NO18: C, 56.01; H, 4.31. Found: C, 55.88;
H, 4.35.
2,3,4,6-Tetra-O-benzoyl-h-
D
-mannopyranosyl-(1“3)-
4,6-O-benzylidene-1,2-O-ethylidene-i-
D
-mannopyranose
(3).—To a cooled solution (0 °C) of 1 (7.40 g, 10
mmol) and 2 (R-form, 2.94 g, 10 mmol) in anhyd
CH2Cl2 (50 mL) was added TMSOTf (50 mL, 0.28
mmol). The mixture was stirred at this temperature for
2 h, and then quenched with Et3N (2 drops). The
solvents were evaporated in vacuo to give a residue,
which was purified by silica gel column chromatogra-
phy (2:1 petroleum ether–EtOAc) to give disaccharide
3 as white foam (4.72 g, 83%): For R-isomer: [h]D
1
−80.6° (c 1.0, CHCl3); H NMR (400 MHz, CDCl3): l
8.07–7.37 (m, 25 H, PhH), 6.07 (dd, 1 H, J3,4=J4,5 9.8
Hz, H-4%), 6.01 (dd, 1 H, J2,3 3.0, J3,4 9.8 Hz, H-3%), 5.86
(dd, 1 H, J1,2 1.0, J2,3 3.0 Hz, H-2%), 5.66 (s, 1 H,
PhCH), 5.59 (d, 1 H, J1,2 1.0 Hz, H-1%), 5.41 (q, 1 H, J
4.7 Hz, MeCH), 5.07 (d, 1 H, J1,2 1.6 Hz, H-1), 4.69
(dd, 1 H, J2,3 2.0, J3,4 9.7 Hz, H-3), 4.62 (ddd, 1 H, J4,5
9.7, J5,6%a 4.2, J5,6%b 2.6 Hz, H-5%), 4.70 (dd, 1 H, J5,6% 4.5,
J6,6% 13.5 Hz, H-6%), 4.34–4.23 (m, 4 H, H-2, H-6% and 2
H-6), 3.81 (dd, 1 H, J3,4=J4,5 9.7 Hz, H-4), 3.36 (ddd,
1 H, J4,5 9.7, J5,6a 4.5, J5,6b 4.3 Hz, H-5), 1.58 (d, 3 H,
J 4.7 Hz, MeCH). Anal. Calcd for C49H44O15: C 67.42;
H 5.08. Found: C 67.21; H 5.06.
Allyl 3,4,6-tri-O-benzoyl-h-
D
-mannopyranosyl-(1“
2)-3,4,6-tri-O-benzoyl-h- -mannopyranoside (10).—To
D
a solution of 9 (8.67 g, 10.0 mmol) in anhyd MeOH
(100 mL) and CH2Cl2 (50 mL) was added AcCl (0.5
mL). The flask was stoppered, and the solution was
stirred at rt until TLC (3:1 petroleum ether–EtOAc)
showed that the starting material disappeared (about
0.5–1 h). The solution was neutralized with Et3N, then
concentrated to dryness. The residue was passed
through a short silica gel column (3:1 petroleum ether–
EtOAc) to give 10 (6.69 g, 80%) as a white solid: [h]D
1
−14.2° (c 1.0, CHCl3); H NMR (400 MHz, CDCl3): l
2,3,4,6-Tetra-O-benzoyl-h-
D
-mannopyranosyl-(1“3)-
8.14–7.24 (m, 30 H, 6 BzH), 5.97 (dd, 1 H, J3,4=J4,5
9.9 Hz, H-4%), 5.90 (dd, 1 H, J3,4=J4,5 10.0 Hz, H-4),
5.88 (m, 1 H, CHꢀCH2), 5.82 (dd, 1 H, J2,3 3.2, J3,4 9.9
Hz, H-3%), 5.79 (dd, 1 H, J2,3 3.1, J3,4 10.0 Hz, H-3),
5.26 (dd, 1 H, J 1.5, J 17.2 Hz, CHꢀCH2), 5.18 (dd, 1
H, J 1.5, J 10.6 Hz, CHꢀCH2), 5.17 (d, 1 H, J1,2 1.4 Hz,
H-1%), 5.14 (d, 1 H, J1,2 1.6 Hz, H-1), 4.63–4.35 (m, 8
H, H-2%, H-2, H-5%, H-5, 2 H-6%, 2 H-6), 4.18 (dd, 1 H,
J 5.4, J 12.7 Hz, CH2ꢁCHꢀCH2), 3.92 (dd, 1 H, J 6.0,
J 12.7 Hz, CH2ꢁCHꢀCH2). Anal. Calcd for C57H50O17:
C, 67.99; H, 5.00. Found: C, 68.12; H, 5.03.
2,4,6-tri-O-acethyl-h- -mannopyranosyl trichloroacet-
D
imidate (7).—To a solution of 3 (7.58 g, 10.0 mmol) in
anhyd MeOH (250 mL) and CH2Cl2 (50 mL) was
added AcCl (1.5 mL). The flask was stoppered, and the
solution was stirred at rt for 1 h, at the end of which
time TLC (1:2 petroleum ether–EtOAc) showed that
the starting material had disappeared. The solution was
neutralized with Et3N, then concentrated to dryness.
The residue was washed with water and extracted 3–4
times with CH2Cl2. The organic phase was dried over
anhyd Na2SO4, then concentrated to dryness. The
residue was passed through a short silica gel column
(1:1.5 petroleum ether–EtOAc) to give 4 (6.86g, 90%)
as a white solid. The white solid was dissolved in
pyridine (40 mL), and then Ac2O (20 mL) was added.
After stirring the mixture at rt for 12 h, TLC (3:2
petroleum ether–EtOAc) indicated that the reaction
was complete. The reaction mixture was concentrated
to dryness. The resultant crude product 5 was dissolved
in a M solution of NH3–MeOH (400 mL) and stirred
at rt until TLC (3:1 petroleum ether–EtOAc) indicated
that the reaction was complete. The solution was con-
centrated to give compound 6 as a syrup. A mixture of
6, trichloroacetonitrile (3.2 mL, 15 mmol), and 1,8-di-
2,3,4,6-Tetra-O-benzoyl-h-
2,4,6-tri-O-acetyl-h- -mannopyranosyl-(1“2)-3,4,6-
tri-O-benzoyl-h- -mannopyranosyl-(1“2)-3,4,6-tri-O-
benzoyl-h- -mannopyranosyl trichloroacetimidate (13).
D
-mannopyranosyl-(1“3)-
D
D
D
—Compound 11 (3.25 g, 61%) was prepared by cou-
pling of 7 (2.91 g, 2.83 mmol) with 10 (2.84 g, 2.82
mmol) under the same conditions as described for the
synthesis of 3 by coupling of 1 with 2. To a solution of
11 (0.8 g, 0.43 mmol) in anhyd MeOH (20 mL) was
added PdCl2 (50 mg). After stirring the mixture at rt for
2 h, TLC (3:2 petroleum ether–EtOAc) indicated that
the reaction was complete. The mixture was filtered,
and the solution was concentrated to dryness. The
resultant residue was purified by flash chromatography

1370
Y. Zeng et al. / Carbohydrate Research 337 (2002) 1367–1371
(2:1 petroleum ether–EtOAc) to give 13 (0.6 g, 70% for
two steps) as a white foam: [h]D −19.6° (c 1.0, CHCl3);
1H NMR (400 MHz, CDCl3): l 8.69 (s, 1 H, NH),
8.15–7.21 (m, 50 H, 10 BzH), 6.59 (d, 1 H, J1,2 2.3 Hz,
H-1), 6.24 (dd, 1 H, J3,4=J4,5 10.1 Hz, H-4%%%), 6.05 (dd,
1 H, J3,4=J4,5 10.3 Hz, H-4%), 5.97 (dd, 1 H, J3,4=J4,5
10.1 Hz, H-4%%), 5.81 (dd, 1 H, J2,3 3.2, J3,4 10.1 Hz,
H-3%%%), 5.78 (dd, 1 H, J2,3 3.1, J3,4 10.3 Hz, H-3%), 5.73
(dd, 1 H, J2,3 3.2, J3,4 10.1 Hz, H-3%%), 5.47–5.39 (m, 3
H, H-1%%%, H-1%%, H-3), 5.30 (d, 1 H, J1,2 1.6 Hz, H-1%),
5.23 (dd, 1 H, J3,4=J4,5 12.0 Hz, H-4), 4.71–4.34 (m,
13 H, H-2%%%, H-2%%, H-2%, H-2, H-5%%%, 2 H-6%%%, 2 H-6%%, 2
H-6%, 2 H-6), 3.96 (m, 3 H, H-5%%, H-5%, H-5), 2.22 (s, 3
H, MeCO), 2.04 (s, 3 H, MeCO), 2.02 (s, 3 H, MeCO).
Anal. Calcd for C102H88Cl3NO34: C, 61.93; H, 4.48.
Found: C, 61.78; H, 4.43.
Allyl
(1“3)-2,4,6-tri-O-acethyl-h-
3,4,6-tri-O-benzoyl-h- -mannopyranosyl-(1“2)-3,4,6-
tri-O-benzoyl-h- -mannopyranosyl-(1“6)-2,3,4-tri-O-
benzoyl-h- -mannopyranosyl-(1“2)-3,4,6-tri-O-ben-
zoyl-h- -mannopyranosyl-(1“2)-3,4,6-tri-O-benzoyl-
h- -mannopyranoside (17).—Under the same condi-
2,3,4,6-tetra-O-benzoyl-h-
D-mannopyranosyl-
D
-mannopyranosyl-(1“2)-
D
D
D
D
D
tions as described for the synthesis of 3 by coupling of
1 with 2, heptasaccharide 17 (94 mg, 51%) was obtained
from coupling of 13 (150 mg, 0.076 mmol) with 16 (100
mg, 0.067 mmol): [h]D −19.6° (c 1.0, CHCl3); 1H
NMR (400 MHz, CDCl3): l 8.19–7.20 (m, 95 H, 9
BzH), 6.31–6.18 (m, 2 H, 2 H-4), 6.08–5.63 (m, 11 H,
H-2, 4 H-3, 5 H-4, CHꢀCH2), 5.51–5.15 (m, 10 H, 4
H-1, H-2, 3 H-3, 2 CHꢀCH2), 4.81–4.33 (m, 22 H, 3
H-1, 3 H-2, 4 H-5, 12 H-6), 4.30–4.08 (m, 5 H, 2 H-5,
2 H-6, CH2ꢁCHꢀCH2), 4.06–3.68 (m, 4 H, 2 H-2, H-5,
CH2ꢁCHꢀCH2), 2.19 (s, 3 H, MeCO), 1.94 (s, 3 H,
MeCO), 1.85 (s, 3 H, MeCO). 13C NMR (100 MHz,
CDCl3): l 170.1, 169.8, 169.6 (3 C, 3 MeCO), 166.2,
165.9, 165.8, 165.7, 165.6, 165.5, 165.3, 165.2, 165.1,
165.0, 164.9 (19 C, PhCO), 133.7–132.2, 130.2–129.2,
129.1–127.9 (PhCO, ꢁCH2ꢁCHꢀCH2), 118.0 (1 C,
ꢁCH2ꢁCHꢀCH2), 100.6, 99.8, 99.8, 99.7, 99.0, 98.3,
98.0 (7 C-1), 77.3 (C-3%%%%%), 73.4, 72.2, 71.3, 70.8, 70.6,
70.3, 70.1, 70.0, 69.7, 69.6, 69.4, 69.2, 68.7, 68.5, 67.2,
66.3, 65.9, 65.8, 63.6, 63.5, 62.9, 62.8, 62.2, 62.0 (C-2–
C-6, ꢁCH2ꢁCHꢀCH2), 21.4, 20.7, 20.5 (3 MeCO). Anal.
Calcd for C184H158O58: C, 67.03; H, 4.83. Found: C,
67.10; H, 4.82.
Allyl 2,3,4-tri-O-benzoyl-h-
2)-3,4,6-tri-O-benzoyl-h- -mannopyranosyl-(1“2)-
3,4,6-tri-O-benzoyl-h- -mannopyranoside (16).—To a
D
-mannopyranosyl-(1“
D
D
cooled solution (0 °C) of 10 (1.01 g, 1 mmol) and 14
(0.81g, 1.2 mmol) in anhyd CH2Cl2 (50 mL) was added
TMSOTf (30 mL, 0.12 mmol). The mixture was stirred
at 0 °C for 2 h and then quenched with Et3N (2 drops).
The solvent was evaporated to give a residue, which
was purified by silica gel column chromatography (2:1
petroleum ether–EtOAc) to give trisaccharide 15 as a
foamy solid (1.24 g, 82%). Compound 15 was dissolved
in anhyd MeOH (200 mL) and CH2Cl2 (100 mL), and
to the mixture was added AcCl (1.2 mL). The flask was
stoppered, and the solution was stirred at rt until TLC
(1:1 petroleum ether–EtOAc) showed that the starting
material disappeared (2 h). The solution was neutral-
ized with Et3N, then concentrated to dryness. The
residue was passed through a short silica gel column
(1:2 petroleum ether–EtOAc) to give 16 (810 mg, 67%
for two steps) as a white solid: [h]D −71.5° (c 1.0,
Allyl h-
osyl-(1“2)-h-
pyranosyl - (1“6) - h -
mannopyranosyl-(1“2)-h-
D
-mannopyranosyl-(1“3)-h-
-mannopyranosyl-(1“2)-h-
- mannopyranosyl - (1“2) - h -
-mannopyranoside (18).—
D
-mannopyran-
D
D
-manno-
D
D-
D
Compound 17 (75 mg, 0.0275 mmol) was dissolved in
satd NH3–MeOH (10 mL). After 2 weeks at rt, the
reaction solution was concentrated, and the residue was
purified on a BioGel P2 column with MeOH–water as
the eluent to afford 18 (24 mg, 80%) as a pulverous
1
CHCl3); H NMR (400 MHz, CDCl3): l 8.14–7.24 (m,
45 H, 9 BzH), 6.04–5.86 (m, 5 H, H-3%%, H-4, H-4%,
H-4%%, CHꢀCH2), 5.71–5.65 (m, 3 H, H-3, H-3%,H-2%%),
5.39 (d, 1 H, J1,2 1.5 Hz, H-1%%), 5.27 (dd, 1 H, J 1.4, J
17.2 Hz, CHꢀCH2), 5.18 (dd, 1 H, J 1.4, J 10.3 Hz,
CHꢀCH2), 5.14 (d, 1 H, J1,2 1.9 Hz, H-1%), 4.73 (d, 1 H,
J1,2 1.0 Hz, H-1), 4.63–4.50 (m, 6 H, H-5%%, 2 H-6%%, 2
H-6%, H-6),. 4.41–4.36 (m, 2 H, H-5%, H-6), 4.18 (dd, 1
H, J 6.0, J 12.7 Hz, CH2ꢁCHꢀCH2), 4.11 (ddd, 1 H, J4,5
11.3, J5,6a 8.8, J5,6b 4.4 Hz, H-5), 3.96 (dd, 1 H, J 6.0, J
12.7 Hz, CH2ꢁCHꢀCH2), 3.52 (m, 2 H, H-2%, H-2). 13C
NMR (100 MHz, DCCl3): l 166.2, 166.2, 166.0, 166.0,
165.4, 165.2, 165.1, 164.9, 164.7 (9 C, 9 PhCO), 133.5–
1
crystalloid: [h]D +52.6° (c 1.0, water); H NMR (400
MHz, D2O): l 5.87 (m, 1 H, CHꢀCH2), 5.25 (dd, 1 H,
J 1.6, J 17.2 Hz, CHꢀCH2), 5.20, 5.18, 5.10, 5.05, 5.04
(5 H, H-1), 5.01 (dd, 1 H, J 1.6, J 10.8 Hz, CHꢀCH2),
4.94, 4.94 (2 H, H-1), 4.18–3.40 (m, 44 H,
CH2ꢁCHꢀCH2, H-2–H-6). 13C NMR (100 MHz, D2O):
l
132.7 (1 C, ꢁCH2ꢁCHꢀCH2), 117.9 (1 C,
ꢁCH2ꢁCHꢀCH2), 101.7, 101.7, 101.6, 100.3, 100.0, 97.7,
96.9 (7 C-1, JC-1,H-1 172.0–173.5 Hz), 78.2, 78.1, 78.0,
77.8, 77.4, 75.4, 75.2, 75.0, 73.6, 72.9, 72.8, 72.8, 72.4,
72.3, 72.2, 71.5, 71.1, 70.2, 70.0, 70.0, 69.8, 69.7, 69.5,
69.4, 69.1, 67.6, 66.5, 66.4, 66.3, 65.8, 65.6, 61.9, 60.5,
60.4, 60.2, 60.2 (C-2–C-6, ꢁCH2ꢁCHꢀCH2). Anal.
Calcd for C45H76O36: C, 45.30; H, 6.42. Found: C,
45.17; H, 6.47.
132.6,
130.0–128.2
(PhCO),
118.0
(1
C,
ꢁCH2ꢁCHꢀCH2), 100.4, 99.7, 98.0 (3 C, 3 C-1), 71.9,
71.9, 70.3, 70.0, 69.6, 69.3, 68.7, 68.7, 68.6, 67.3, 67.0,
66.7, 63.5, 63.5, 61.6, 61.4 (C-2–C-6, ꢁCH2ꢁCHꢀCH2).
Anal. Calcd for C84H72O25: C, 68.10; H, 4.90. Found:
C, 67.90; H, 4.87.

Y. Zeng et al. / Carbohydrate Research 337 (2002) 1367–1371
1371
T.; Shibata, N.; Suzuki, S. Arch. Biochem. Biophys. 1992,
294, 662–669.
Acknowledgements
7. Schmidt, R. R.; Kinzy, W. Ad6. Carbohydr. Chem.
Biochem. 1994, 50, 21–125.
8. (a) Zhu, Y.; Kong, F. Synlett 2000, 1783–1788;
(b) Zhu, Y.; Kong, F. Chin. J. Chem. 2001, 19, 119–123.
9. (a) Auzanneau, F.-I.; Forooghian, F.; Pinto, B. M. Car-
bohydr. Res. 1996, 291, 21–40;
This work was supported by The Chinese Academy
of Sciences (Projects KJ952J1510 and KIP-
RCEES9904) and by The National Natural Science
Foundation of China (Projects 39970864 and
30070815).
(b) Wang, W.; Kong, F. Carbohydr. Res. 1999, 315,
128–135.
10. Ogawa, T.; Yamamoto, H. Carbohydr. Res. 1985, 137,
79–87.
References
11. Heng, L.; Ning, J.; Kong, F. J. Carbohydr. Chem. 2001,
20, 285–296.
1. Goodman, D. S.; Teplitz, E. D.; Wishner, A.; Klein, R.
S.; Burk, P. G.; Hershenbaum, E. J. Am. Acad. Dermatol.
1987, 17, 210–218.
12. (a) Nakahara, Y.; Shibayama, S.; Nakahara, Y.; Ogawa,
T. Carbohydr. Res. 1996, 280, 67–84;
2. Bodey, G. P. Am. J. Med. 1984, 77, 13–22.
3. Spebar, M. J.; Pruitt, B. A. J. Trauma 1981, 21, 237–243.
4. Silverman, S., Jr.; Luangjarmekorn, L.; Greenspan, D. J.
Oral Med. 1984, 39, 194–201.
(b) Grice, P.; Ley, S. V.; Pietrunzka, J. Angew. Chem.,
Int. Ed. Engl. 1996, 35, 197–200;
(c) Ogawa, T.; Sasajima, K. Tetrahedron 1981, 37, 2787–
2792.
5. Goldman, W. E. Science 1999, 285, 539–541.
6. Kobayashi, H.; Mitobe, H.; Takahashi, K.; Yamamoto,
13. Betaneli, V. I.; Ovchinnikov, M. V.; Kochetkov, N. K.
Carbohydr. Res. 1982, 107, 285–291.