A practical synthesis of a (1→6)-linked β-D-glucosamine nonasaccharide

Article abstract of DOI:10.1016/S0008-6215(02)00494-9

A (1→6)-β-D-glucosamine nonasaccharide was convergently synthesized using isopropyl thioglycosides as donors. Anomeric acetylated glucosamine derivatives were proved to be good acceptors in the NIS/TMSOTf catalyzed glycosylation. The target nonasaccharide showed a mild antitumor activity against H₂₂ on the preliminary mice tests.

Full text of DOI:10.1016/S0008-6215(02)00494-9

Carbohydrate Research 338 (2003) 495–502
www.elsevier.com/locate/carres
A practical synthesis of a (1“6)-linked b- -glucosamine
D
nonasaccharide
Feng Yang, Yuguo Du*
Research Center for Eco-En6ironmental Sciences, Chinese Academy of Sciences, Academia Sinica, P.O. Box 2871, Beijing 100085, PR China
Received 18 September 2002; accepted 16 November 2002
Abstract
A (1“6)-b-D-glucosamine nonasaccharide was convergently synthesized using isopropyl thioglycosides as donors. Anomeric
acetylated glucosamine derivatives were proved to be good acceptors in the NIS/TMSOTf catalyzed glycosylation. The target
nonasaccharide showed a mild antitumor activity against H22 on the preliminary mice tests. © 2003 Elsevier Science Ltd. All
rights reserved.
Keywords: Glycosylation; Oligosaccharides; Glucosamine; Bioactivity
1. Introduction
the first synthesis of
nonasaccharide and preliminary results of its anticancer
activity against the H22 tumor.
a
(1“6)-b-D-glucosamine
2-Amino-2-deoxy-
D
-glucose (glucosamine) is the most
common amino sugar and occurs as a constituent of
glycoproteins, glycolipids, bacterial lipopolysaccha-
rides, proteoglycans and nodulation factors associated
with leguminous plants.1 Chitin and chitosan, poly-
2. Results and discussion
(1“4)-b-D-glucosamine derivatives, are widely used as
The convergent synthesis of the target nonasaccharide
is outlined in Scheme 1. As we have observed in previ-
ous work that the rapid consumption of 3,4,6-tri-O-
a bifidus factor to promote animal growth and as an
elicitor to protect plants from fungal invasions.2 Glu-
cosamine oligosaccharides with the (1“6)-linkage have
acetyl-2-deoxy-2-phthalimido-a-D-glucopyranosyl
gotten less attention with respect to their b-
D
-(1“4)-
trichloroacetimidate during its glycosylation afforded a
linked counterparts.3 A literature search suggests that
low yield of product,7 we decided to use isopropyl
(1“6)-b-D-glucosamine oligosaccharides may act as a
3,4-di-O-acetyl-2-deoxy-2-phthalimido-1-thio-b-D-
potential antitumor and immunostimulating agents.4
More impressively, a carbohydrate-based catalyst, con-
glucopyranoside (7) as the basic building block for
preparing the elongation synthon. Thus, the b isomer of
4,8 re-crystallized from its a,b mixture, was reacted with
2-propanethiol under the promotion of BF3·Et2O in
CH2Cl2 at room temperature to give thioglycoside 5 in
86% yield. Using the a,b mixture of 4, we could also get
a good yield of 5 in CH2Cl2 under reflux. Saponifica-
tion of 5 with NaOMe in MeOH, followed by 6-O-sily-
lation with tert-butylchlorodimethylsilane in pyridine
and subsequent in situ acetylation furnished key syn-
thon 7.
taining three (1“6)-linked b-D-glucosamine units, has
been designed and recently synthesized.5 It showed a
marked rate enhancement and specificity for the hy-
drolysis of GTP to GDP and orthophosphate (OP). We
proved, in a test in mice, that the b-D-glucosamine
hexamer could significantly increase the number of
white blood cells and marrow cells compared to the
results from chemotherapy (CTX).6 Herein, we report
We were gratified to find that the coupling of donor
7 with the acetyl acceptor, 1,3,4-tri-O-acetyl-2-deoxy-2-
* Corresponding author. Tel.: +86-10-62914475; fax: +
86-10-62923563
E-mail address: ygdu@mail.rcees.ac.cn (Y. Du).
phthalimido-a,b- -glucopyranose (3), in CH2Cl2 in the
D
0008-6215/03/$ - see front matter © 2003 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(02)00494-9

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F. Yang, Y. Du / Carbohydrate Research 338 (2003) 495–502
presence of NIS/TMSOTf at −20 °C gave an excellent
yield of disaccharide 10. This result significantly sim-
plified our synthesis towards the target. Removal of the
silyl group from 10 with BF3·Et2O (“11), followed by
glycosylation with thioglycoside 5, afforded fully O-
acetylated triglucosamine derivative 12. It should be
noted that the TBAF-desilylation caused acyl migration
from C-4 to C-6 in this case.9 A modified Helfrich
reaction of 12 and 2-propanethiol in CH2Cl2 under
reflux gave isopropyl thioglycoside 13 in 73% yield. If
the above reaction was carried out at room tempera-
ture, compound 13 was obtained in 53% yield, and the
a isomer of 12 was recovered. Condensation of trisac-
charide donor 13 with disaccharide acceptor 11 gave
pentasaccharide 14 with the predominant a configura-
tion on the reducing end (sugar unit I). Pure a product
was obtained in a yield of 78% after a silica gel column
separation. Again, a modified Helfrich reaction of pen-
tasaccharide 14 and 2-propanethiol in CH2Cl2 under
reflux gave isopropyl thioglycoside 15 (60%). No reac-
tion was observed at 0 °C in 2 h.
Convergently, glycosylation of 7 with octanol in
CH2Cl2 using NIS/TMSOTf catalysts gave glycoside 8,
which was further desilylated with 2 equiv of BF3·Et2O
to give acceptor 9 (85% from 7). Reiterative coupling
and desilylation of 7 and 9 finally gave a tetra-
saccharide acceptor 21 in 28% overall yield (from 9 to
21). With pentasaccharide donor 15 and tetrasaccharide
acceptor 21 in hand, we tried condensation of these two
species in CH2Cl2 at −10 °C using NIS/TMSOTf pro-
Scheme 1. Reagents and conditions: (a) TBSCl, Pyr; Ac2O; 70% for 2; 78% for 7; (b) BF3·Et2O, CH2Cl2; 99.5% for 3; 89% for
9; 84.6% for 11; 78% for 17; 86.5% for 19; 84% for 21; (c) Ac2O, Pyr; (d) BF3·Et2O, CH2Cl2, 86%; (e) NIS, TMSOTf, CH2Cl2;
95.5% for 8; 94% for 10; 86% for 12; 78% for 14; 80% for 16; 92% for 18; 67.5% for 20; 47.4% for 24; (f) BF3·Et2O, CH2Cl2, reflux;
73% for 13; 60% for 15; (g) MeOH satd with NH3; 72%.

F. Yang, Y. Du / Carbohydrate Research 338 (2003) 495–502
497
moters, and 47.4% of nonasaccharide 24 was obtained.
Full deprotection of 24 in ammonia-saturated methanol
afforded nonasaccharide 25 in good yield (72%).
product was subjected to column chromatography (1:1
EtOAc–petroleum ether) to give syrupy 3 (1.97 g, 70%)
1
as an a,b mixture (3:2): H NMR for b isomer: l 1.87,
The antitumor activity of nonasaccharide 25 was
preliminarily studied according to the method described
by Sasaki and co-workers.10 Balb/c mice weighing
about 20 g and H22 (2.5×107 cells) were used for the
bioassay. Lentinan (imported from Japan for medical
usage) and Cisplatin® (CDDP) were selected as the
positive controls in parallel tests. The samples were
injected daily for 9 days, while CDDP was given every
other day. The tumor inhibition ratios for 25, lentinan
and CDDP were 36.8–43.1% (dosage: 0.5–2.0 mg/kg/
day), 37.3% (dosage: 1 mg/kg/day) and 55.4–56.1%
(dosage: 3 mg/kg/every other day), respectively.
2.00, 2.08 (3 s, 9 H, Ac), 3.62–3.85 (m, 3 H, H-6a,
H-6b, H-5), 4.45 (dd, 1 H, J 10.4, 8.8 Hz, H-2), 5.17
(dd, 1 H, J 9.2, 5.8 Hz, H-4), 5.93 (dd, 1 H, J 10.4, 9.2
Hz, H-3), 6.53 (d, 1 H, J 8.8 Hz, H-1), 7.25–7.87 (m, 4
1
H, Ph); H NMR for a isomer: l 1.89, 2.09, 2.10 (3 s,
9 H, Ac), 3.62–3.84 (m, 2 H, H-6a, H-6b), 4.12–4.14
(m, 1 H, H-5), 4.70 (dd, 1 H, J 11.6, 3.4 Hz, H-2), 5.15
(dd, 1 H, J 9.2, 3.0 Hz, H-4), 6.30 (d, 1 H, J 3.4 Hz,
H-1), 6.61 (dd, 1 H, J 11.6, 9.2 Hz, H-3), 7.26–8.02 (m,
4 H, Ph). MALDITOF-MS: Calcd for C20H21NO10: 435
[M]; Found 458.3 [M+Na]+.
In conclusion, we have described a practical conver-
3.3. Isopropyl 3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-
gent synthesis of the linear (1“6)-b-
D-glucosamine
1-thio-b-D-glucopyranoside (5)
nonasaccharide. The reiterative usage of 6-O-silylated
isopropyl thioglycoside and C-1 acetylated acceptor
simplified the whole procedure. Moreover, the synthetic
target compound showed a mild antitumor activity
towards liver cancer model H22.
To a mixture of b anomer of 4 (9.53 g, 20 mmol) and
2-propanethiol (2.43 mL, 26 mmol) in CH2Cl2 (50 mL)
was added BF3·Et2O (3.25 mL, 26 mmol). The mixture
was stirred at rt for 2 h, then poured into cold satd aq
NaHCO3, and extracted with CH2Cl2. The organic
phase was concentrated, and the crude product was
subjected to column chromatography (2:5 EtOAc–
petroleum ether) to give syrupy 5 (8.47 g, 86%): [h]D
3. Experimental
1
3.1. General methods
+42° (c 1, CHCl3); H NMR: l 1.24 (t, 6 H, J 7.2 Hz,
SCH(CH3)2), 1.87, 2.05, 2.11 (3 s, 9 H, Ac), 3.13–3.17
(m, 1 H, SCH(CH3)2), 3.90–3.92 (m, 1 H, H-5), 4.17
(dd, 1 H, J 12.0, 2.2 Hz, H-6a), 5.31 (dd, 1 H, J 12.0,
4.3 Hz, H-6b), 4.37 (t, 1 H, J 10.0 Hz, H-2), 5.17 (t, J
10.0 Hz, H-4), 5.57 (d, 1 H, J 10.0 Hz, H-1), 5.83 (t, 1
H, J 10.0 Hz, H-3), 7.74–7.88 (m, 4 H, Ph). Anal.
Calcd for C23H27NO9S: C, 55.97; H, 5.51. Found: C,
55.80; H, 5.67.
Optical rotations were determined at 25 °C with a
Perkin–Elmer Model 241-Mc automatic polarimeter.
1H NMR, 13C NMR and H–1H, H–13C COSY spec-
tra were recorded with a Bruker ARX 400 spectrome-
ters for solutions in CDCl3 or D2O. Chemical shifts are
given in ppm downfield from internal Me4Si. Mass
spectra were measured using MALDITOF-MS with
a-cyano-4-hydroxycinnamic acid (CCA) as matrix.
Thin-layer chromatography (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
detector.
1
1
3.4. Isopropyl 3,4-di-O-acetyl-6-O-tert-butyldimethylsil-
yl-2-deoxy-2-phthalimido-1-thio-b-D-glucopyranoside (7)
Compound 5 (10 g, 20.26 mmol) in MeOH (120 mL)
was treated with 1 M NaOMe (5 mL) at rt for 2 h and
then neutralized with Amberlite IR-120 (H+) resin.
After filtration, the filtrate was concentrated. The crude
3.2. 1,3,4-Tri-O-acetyl-2-deoxy-2-phthalimido-D-
glucopyranose (3)
1
6 gave the following H NMR: l 1.18 (2 t, 6 H, J 6.4
To a pre-cooled solution of compound 18a (2.0 g, 6.467
mmol) in pyridine (10 mL) was added tert-
butylchlorodimethylsilane (1.065 g, 7.1 mmol) and
DMAP (20 mg). The mixture was stirred at room
temperature (rt) for 24 h, and then a premixed solution
of acetic anhydride (6 mL) and pyridine (6 mL) was
added. After 4 h, the mixture was co-evaporated to
dryness with the help of toluene. The above crude
product was treated with BF3·Et2O (1.0 mL, 7.96
mmol) in CH2Cl2 (25 mL) at rt for 75 min, then poured
into cold satd aq NaHCO3, and extracted with CH2Cl2.
The organic phase was concentrated, and the crude
Hz, CH3), 3.09–3.13 (m, 1 H, CH), 3.48–3.50 (m, 1 H,
H-5), 3.68 (t, 1 H, J 10.0 Hz, H-4), 3.88 (d, 2 H, J 2.8
Hz, H-6), 4.11 (t, 1 H, J 10.0 Hz, H-2), 4.33 (t, 1 H, J
10.0 Hz, H-3), 5.42 (d, 1 H, J 10.0 Hz, H-1), 7.69–7.82
(m, 4 H, Ph). To the above residue was added tert-
butylchlorodimethylsilane (3.45 g, 23 mmol) and
DMAP (30 mg). The mixture was stirred at rt for 10 h,
then was added a premixed solution of acetic anhydride
(8 mL) and pyridine (8 mL). The mixture was stirred at
rt for 4 h and then co-evaporated with toluene. The
residue was subjected to the column chromatography
(1:1 EtOAc–petroleum ether) to give 7 (8.93 g, 78%) as

498
F. Yang, Y. Du / Carbohydrate Research 338 (2003) 495–502
a foam: [h]D +53° (c 1, CHCl3); 1H NMR: l 0.06, 0.08
(2 s, 6 H, Si(CH3)2), 0.90 (s, 9 H, t-Bu), 1.22, 1.26 (2 s,
6 H, SCH(CH3)2), 1.85, 2.03 (2 s, 6 H, Ac), 3.13–3.16
(m, 1 H, SCH), 3.69–3.72 (m, 3 H, 2 H-6, H-5), 4.34 (t,
J 10.0 Hz, H-2), 5.09 (t, 1 H, J 10.0 Hz, H-4), 5.53 (d,
1 H, J 10.0 Hz, H-1), 5.82 (t, 1 H, J 10.0, H-3),
7.27–7.87 (m, 4 H, Ph); 13C NMR: l −5.40, 18.27,
20.48, 20.71, 23.76, 24.12, 25.81, 35.23, 54.04 (C-2),
62.81 (C-6), 69.63 (C-4), 71.97 (C-3), 78.99 (C-5), 80.45
(C-1), 123.60, 123.64, 131.28, 131.67, 134.17, 134.28,
167.17, 167.80, 169.43, 170.23. MALDITOF-MS: Calcd
for C27H39NO8SSi: 565 [M]; Found 588.3 [M+Na]+.
column chromatography (1:1 EtOAc–petroleum ether)
to give syrupy 9 (2.9 g, 89%): [h]D +30° (c 1, CHCl3);
1H NMR: l 0.81 (t, 3 H, J 7.2 Hz, CH2CH3), 0.88–1.43
(m, 12 H, 6 CH2), 1.88, 2.01 (2 s, 6 H, Ac), 3.42–3.45
(m, 1 H, OCHaHb), 3.64–3.82 (m, 3 H, H-5, H-6a,
H-6b), 3.82–3.85 (m, 1 H, OCHaHb), 4.29 (dd, 1 H, J
10.8, 8.4 Hz, H-2), 5.12 (t, 1 H, J 9.3 Hz, H-4), 5.39 (d,
1 H, J 8.4 Hz, H-1), 5.84 (dd, 1 H, J 10.8, 9.3 Hz, H-3),
7.34–7.87 (m, 4 H, Ph). Anal. Calcd for C26H35NO9: C,
61.77; H, 6.98. Found: C, 61.54; H, 7.09.
3.8. 3,4-Di-O-acetyl-6-O-tert-butyldimethylsilyl-2-de-
oxy-2-phthalimido-b-
O-acetyl-2-deoxy-2-phthalimido-
D
-glucopyranosyl-(1“6)-1,3,4-tri-
3.5. General procedure for NIS/TMSOTf-catalyzed
coupling reaction
D-glucopyranose (10)
Compound 7 (885 mg, 1.56 mmol) and 3 (620 mg, 1.42
To a solution of isopropyl thioglycoside donor (1
mmol) and alcohol acceptor (0.98 mmol or as claimed
specifically) in anhyd CH2Cl2 at −20 °C was added
NIS (1.5 mmol) and TMSOTf (0.05 mmol), respec-
tively, under N2 protection. The mixture was stirred
under these conditions for 60 min, neutralized with
Et3N and then concentrated. The residue was subjected
to silica gel column chromatography (EtOAc–
petroleum ether) to give the desired product.
mmol) was coupled as described in general procedure to
1
give 10 (1.24 g, 94%): H NMR: l 0.08, 0.09 (2 s, 6 H,
Si(CH3)2), 0.91 (s, 9 H, t-Bu), 1.77, 1.82, 1.84, 1.88,
2.02 (5 s, 5×1.2 H, b-COCH3), 1.71, 1.78, 1.85, 1.93,
2.02 (5 s, 5×1.8 H, a-COCH3), 3.59–3.78 (m, 4 H, 4
H-6), 3.82–3.84 (m, 0.4 H, Hb-5), 3.90 (t, 1 H, J 12.6
Hz, H-5%), 4.12–4.14 (m, 0.6 H, Ha-5), 4.30–4.33 (m,
1.4 H, H-2%, Hb-2), 5.54 (dd, 0.6 H, J 11.6, 3.4 Hz,
Ha-2), 4.90 (t, 0.6 H, J 9.2 Hz, Ha-4), 5.02 (t, 0.4 H, J
10.0 Hz, Hb-4), 5.11 (t, 1 H, J 9.6 Hz, H-4%), 5.41 (d, 1
H, J 8.8 Hz, H-1%), 5.75 (br t, 1.4 H, J 10.0 Hz, H-3%,
Hb-3), 5.94 (d, 0.6 H, J 3.6 Hz, Ha-1), 6.34 (d, 0.4 H, J
8.8 Hz, Hb-1), 6.39 (dd, 0.6 H, J 11.6, 9.2 Hz, Ha-3),
7.69–7.86 (m, 8 H, Ph). MALDITOF-MS: Calcd for
C44H52N2O18Si: 924 [M]; Found 947.1 [M+Na]+.
3.6. Octyl 3,4-di-O-acetyl-6-O-tert-butyldimethylsilyl-2-
deoxy-2-phthalimido-b-D-glucopyranoside (8)
Compound 7 (4.16 g, 7.35 mmol) was reacted with
octanol (1.87 mL, 11.5 mmol) as described in general
procedure to give 8 (4.35 g, 95.5%) as a syrup: [h]D
1
+23° (c 1, CHCl3); H NMR: l 0.06, 0.07 (2 s, 6 H,
Si(CH3)2), 0.79 (t, 3 H, J 7.2 Hz, CH2CH3), 0.89 (s, 9
H, t-Bu), 0.98–1.40 (m, 12 H, 6 CH2), 1.84, 2.01 (2 s,
6 H, Ac), 3.42 (m, 1 H, OCHaHb), 3.67–3.75 (m, 3 H,
H-5, H-6a, H-6b), 3.79–3.81 (m, 1 H, OCHaHb), 4.53
(dd, 1 H, J 10.8, 8.4 Hz, H-2), 5.09 (t, 1 H, J 9.4 Hz,
H-4), 5.31 (d, 1 H, J 8.4 Hz, H-1), 5.79 (dd, 1 H, J 10.8,
9.4 Hz, H-3), 7.71–7.85 (m, 4 H, Ph); 13C NMR: l
−5.47, 13.88, 18.16, 20.33, 20.57, 22.41, 25.69, 28.94,
29.12, 31.48, 54.71, 62.53, 69.59, 69.64, 71.07, 74.61,
97.73, 123.35, 131.36, 134.06, 167.56, 169.30, 170.11.
Anal. Calcd for C32H49NO9Si: C, 62.01; H, 7.97.
Found: C, 61.79; H, 8.05.
3.9. 3,4-Di-O-acetyl-2-deoxy-2-phthalimido-b-
ranosyl-(1“6)-1,3,4-tri-O-acetyl-2-deoxy-2-phthal-
imido- -glucopyranose (11)
D-glucopy-
D
Compound 10 (1.47 g, 1.59 mmol) was treated with
BF3·Et2O (0.4 mL, 3.2 mmol) as described in prepara-
tion of 9 to give 11 (1.09 g, 84.6%) as a foamy solid; H
1
NMR: l 1.80, 1.85, 1.93, 1.94, 2.05 (5 s, 5×1.2 H,
b-COCH3), 1.79, 1.86, 1.93, 1.96, 2.06 (5 s, 5×1.8 H,
a-COCH3), 3.64–3.92 (m, 5.4 H, 4 H-6, Hb-5, H-5%),
4.14–4.16 (m, 0.6 H, Ha-5), 4.31 (dd, 1 H, J 10.0, 8.4
Hz, H-2%), 4.36 (dd, 0.4 H, J 10.0, 8.8 Hz, Hb-2), 5.58
(dd, 0.6 H, J 12.0, 3.4 Hz, Ha-2), 5.00 (t, 0.6 H, J 9.2
Hz, Ha-4), 5.10 (t, 1.4 H, J 10.0 Hz, Hb-4, H-4%), 5.56
(d, 0.4 H, J 8.4 Hz, Hb-1%), 5.41 (d, 0.6 H, J 8.4 Hz,
Ha-1%), 5.78 (t, 1.4 H, J 10.0 Hz, H-3%, Hb-3), 6.00 (d,
0.6 H, J 3.4 Hz, Ha-1), 6.38 (d, 0.4 H, J 8.8 Hz, Hb-1),
6.42 (dd, 0.6 H, J 12.0, 9.2 Hz, Ha-3), 7.7–7.86 (m, 8 H,
Ph). MALDITOF-MS: Calcd for C38H38N2O18: 810
[M]; Found 833.3 [M+Na]+.
3.7. Octyl 3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranoside (9)
Compound 8 (4.0 g, 6.45 mmol) was treated with
BF3·Et2O (1.6 mL, 12.7 mmol) in CH2Cl2 (40 mL) for
15 min at rt, then poured into cold satd aq NaHCO3,
and extracted with CH2Cl2. The organic phase was
concentrated, and the crude product was subjected to

F. Yang, Y. Du / Carbohydrate Research 338 (2003) 495–502
499
3.10. Isopropyl 3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-
b- -glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-
phthalimido-b- -glucopyranosyl-(1“6)-3,4-di-O-acetyl-
2-deoxy-2-phthalimido-1-thio-b- -glucopyranoside (13)
J 8.4 Hz, H-1III), 5.30 (d, 1 H, J 8.4 Hz, H-1IV), 5.49 (d,
1 H, J 8.4 Hz, H-1V), 5.57 (dd, 1 H, J 10.6, 9.2 Hz,
H-3II), 5.61 (dd, 1 H, J 10.6, 9.2 Hz, H-3III), 5.66 (dd,
1 H, J 10.8, J 9.2 Hz, H-3IV), 5.79 (dd, 1 H, J 10.6, 9.2
Hz, H-3V), 5.86 (d, 1 H, J 3.6 Hz, H-1I), 6.39 (dd, 1 H,
J 11.6, 9.2 Hz, H-3I), 7.71–7.92 (m, 20 H, Ph); Selected
13C NMR: l 20.09, 20.12, 20.21, 20.30, 20.34, 20.50,
20.57, 20.72, 52.57, 53.94, 54.15, 61.67, 66.17, 66.97,
67.54, 67.61, 67.85, 68.68, 69.18, 69.26, 69.38, 70.32,
70.47, 70.76, 71.71, 72.40, 72.76, 72.93, 90.05, 97.17,
97.41, 97.72, 97.76, 168.90, 169.01, 169.10, 169.19,
169.22, 169.70, 169.80, 170.41. MALDITOF-MS: Calcd
for C94H91N5O43: 1977.5 [M]; Found 2001.3 [M+Na]+
. Anal. Calcd for C94H91N5O43: C, 57.06; H, 4.64.
Found: C, 56.92; H, 4.67.
D
D
D
Compound 5 (665 mg, 1.35 mmol) and 11 (1.07 g, 1.32
mmol) was coupled as described in general procedure to
give 12 (1.39 g, 86%), which was transformed into the
thioglycoside as described in the preparation of 5 to
1
give 13 (748 mg, 53%): [h]D +43° (c 1, CHCl3); H
NMR: l 1.04, 1.09 (2 s, 6 H, J 6.0 Hz, SCH(CH3)2),
1.78, 1.80, 1.86, 1.93, 1.95, 2.04, 2.16 (7 s, 21 H,
COCH3), 2.97 (m, 1 H SCH(CH3)2), 3.47 (dd, 1 H, J
9.2, 6.8 Hz, H-6a), 3.57–3.60 (m, 1 H, H-5), 3.73–3.85
(m, 3 H, H-6a%, H-5%, H-6b), 3.87–4.10 (m, 2 H, H-5¦,
H-6b%), 4.18–4.23 (m, 3 H, H-2, H-2%, H-6a¦), 4.35 (dd,
1 H, J 10.6, 8.4 Hz, H-2¦), 4.36 (dd, 1 H, H-6b¦), 4.82
(t, 1 H, J 9.2 Hz, H-4), 4.94 (t, 1 H, J 10.4 Hz, H-4%),
5.19 (t, 1 H, J 9.2 Hz, H-4¦), 5.32 (d, 1 H, J 8.8 Hz,
H-1%), 5.39 (d, 1 H, J 10.6 Hz, H-1), 5.52 (d, 1 H, J 8.4
Hz, H-1¦), 5.62 (dd, 1 H, J 10.4, 8.8 Hz, H-3%), 5.68 (dd,
1 H, J 10.0, 9.2 Hz, H-3), 5.77 (dd, 1 H, J 10.6, 9.2 Hz,
H-3¦), 7.71–7.93 (m, 12 H, Ph); 13C NMR: l 20.29,
20.34, 20.34, 20.43, 20.49, 20.56, 20.74, 23.46, 23.86,
34.90, 53.78, 54.38, 61.97, 67.84, 68.21, 68.90, 69.57,
69.61, 70.69, 70.80, 71.40, 72.02, 72.09, 76.76, 80.20,
97.54, 97.93, 123.52, 123.55, 131.70, 131.17, 131.47,
134.117, 134.37, 167.57, 169.26, 169.29, 169.44, 169.49,
169.97, 169.99, 170.03, 170.68. MALDITOF-MS: Calcd
for C59H61N3O25S: 1243 [M]; Found 1266.76 [M+
Na]+. Anal. Calcd for C59H61N3O25S: C, 56.96; H,
4.94. Found: C, 56.81; H, 5.02.
3.12. Isopropyl 3,4,6-tri-O-acetyl-2-deoxy-2-phthal-
imido-b-
oxy-2-phthalimido-b-
acetyl-2-deoxy-2-phthalimido-b-
6)-3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-
pyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-phthal-
imido-1-thio-b- -glucopyranose (15)
D
-glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-de-
-glucopyranosyl-(1“6)-3,4-di-O-
-glucopyranosyl-(1“
-gluco-
D
D
D
D
To a mixture of 14 (1.24 g, 0.627 mmol) and 2-
propanethiol (0.08 mL, 0.86 mmol) in CH2Cl2 (10 mL)
was added BF3·Et2O (0.1 mL, 0.8 mmol). The mixture
was stirred under reflux for 1 h, then poured into cold
satd aq NaHCO3, and extracted with CH2Cl2. The
organic phase was concentrated, and the crude producr
was subjected to column chromatography (5:4 EtOAc–
petroleum ether) to give syrupy 15 (750 mg, 60%): [h]D
1
3.11. 3,4,6-Tri-O-acetyl-2-deoxy-2-phthalimido-b-
glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-ph-
thalimido-b- -glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-
deoxy-2-phthalimido-b- -glucopyranosyl-(1“6)-3,4-di-
O-acetyl-2-deoxy-2-phthalimido-b- -glucopyranosyl-(1
D-
+44° (c 1, CHCl3); H NMR: l 0.99, 1.06 (2 d, 6 H, J
4.6 Hz, SCH(CH3)2), 1.78 (s, 9 H, 3 Ac), 1.79, 1.86,
1.93, 1.94, 1.95, 2.04, 2.17 (7 s, 24 H, 8 Ac), 2.93–2.96
(m, 1 H, SCH(CH3)2), 3.43–3.52 (m, 5 H, H-6aII,
H-6aI, H-6aIII, H-5II, H-5III), 3.56–3.62 (m, 2 H, H-6bI,
H-5I), 3.70–3.80 (m, 4 H, H-6aIV, H-5IV, H-6bII, H-
6bI), 3.90–3.97 (m, 2 H, H-6bIV, H-5V), 4.12–4.24 (m, 5
H, H-2II, H-2III, H-2IV, H-2I, H-6aV), 4.35–4.41 (m, 2
H, H-6bV, H-2V), 4.79–4.85 (m, 3 H, H-4II, H-4III,
H-4IV), 4.93 (t, 1 H, J 9.2 Hz, H-4I), 5.20 (t, 1 H, J 9.2
Hz, H-4V), 5.25 (d, 1 H, J 8.4 Hz, H-1II), 5.30 (d, 1 H,
J 8.4 Hz, H-1III), 5.31 (d, 1 H, J 8.4 Hz, H-1IV), 5.36 (d,
1 H, J 10.4 Hz, H-1I), 5.50 (d, 1 H, J 8.4 Hz, H-1V),
5.61 (dd, 1 H, J 10.6, 9.2 Hz, H-3II), 5.63 (dd, 1 H, J
10.6, 9.2 Hz, H-3III), 5.67 (t, 2 H, J 9.2 Hz, H-3IV,
H-3V), 5.79 (dd, 1 H, J 10.6, 9.2 Hz, H-3I), 7.71–7.92
(m, 20 H, Ph); Selected 13C NMR: l 20.39, 20.43,
20.55, 20.66, 20.82, 23.43, 23.90, 34.81, 53.78 (C-2I),
54.42 (C-2II,C-2III,C-2IV,C-2V), 62.64 (C-6V), 67.19 (C-
6II), 67.85 (C-6III), 67.99 (C-6IV), 68.23 (C-6I), 68.99
(C-4V), 68.48 (C-4II), 69.61 (C-4III, C-4IV), 69.68 (C-4I),
70.66 (C-3II, C-3III, C-3IV), 70.72 (C-3V), 70.82 (C-3I),
71.45 (C-5I), 72.08 (C-5V), 72.59 (C-5IV), 72.65 (C-5III),
D
D
D
“6)-1,3,4-tri-O-acetyl-2-deoxy-2-phthalimido-a-D-
glucopyranose (14)
Coupling of 13 (1.611 g, 1.29 mmol) and 11 (1.02 g,
1.26 mmol) as described in general procedure to give 14
as a foamy solid (1.94 g, 78%): [h]D +61° (c 1, CHCl3);
1H NMR: l 1.77 (×2), 1.78, 1.79, 1.86, 1.89, 1.92,
1.93, 1.94, 1.97, 2.04, 2.16 (11 s, 36 H, COCH3),
3.42–3.58 (m, 5 H, H-6aIII, H-6aI, H-6aII, H-5II, H-5III),
3.62 (d, 1 H, J 12.0 Hz, H-6bII), 3.70–3.85 (m, 3 H,
H-6aIV, H-5IV, H-6bIII), 3.90–3.96 (m, 3 H, H-5V, H-
6bIV, H-6bI), 4.08–4.20 (m, 4 H, H-5I, H-2II, H-6aV,
H-2III), 4.23 (dd, 1 H, J 10.8, 8.4 Hz, H-2IV), 4.32–4.40
(m, 2 H, H-2V, H-6bV), 4.54 (dd, 1 H, J 11.6, 3.6 Hz,
H-2I), 4.76 (t, 1 H, J 9.2 Hz, H-4II), 4.80 (dd, 1 H, J
10.0, 9.2 Hz, H-4III), 4.89 (dd, 1 H, J 10.0, 9.2 Hz,
H-4I), 4.95 (t, 1 H, J 9.2 Hz, H-4IV), 5.19 (t, 1 H, J 9.2
Hz, H-4V), 5.23 (d, 1 H, J 8.4 Hz, H-1II), 5.29 (d, 1 H,

500
F. Yang, Y. Du / Carbohydrate Research 338 (2003) 495–502
73.03 (C-5II), 80.19 (C-1I), 97.43 (C-1IV), 97.56 (C-1III),
97.79 (C-1II), 98.12 (C-1V), 170.77. MALDITOF-MS:
Calcd for C95H95N5O41S: 1993.5 [M]; Found 2016.82
[M+Na]+.
(C-5%), 74.19 (C-5), 97.64 (C-1%), 97.78 (C-1), 123.44,
123.54, 131.37, 131.46, 134.12, 134.18, 169.88, 170.06,
170.10. MALDITOF-MS: Calcd for C44H52N2O17: 880
[M]; Found 903.34 [M+Na]+.
3.13. Octyl 3,4-di-O-acetyl-6-O-tert-butyldimethylsilyl-
2-deoxy-2-phthalimido-b-D-glucopyranosyl-(1“6)-3,4-
di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyranoside
(16)
3.15. Octyl 3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-
glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-
D-
phthalimido-b-D-glucopyranosyl-(1“6)-3,4-di-O-acetyl-
2-deoxy-2-phthalimido-b-
D
-glucopyranoside (19)
Coupling of 7 (1.7 g, 3 mmol) and 9 (1.44 g, 2.85 mmol)
as described in general procedure to give 16 as a syrup
(2.268 g, 80%): [h]D +37° (c 1, CHCl3); H NMR: l
Coupling of 7 (1.87 g, 3.31 mmol) and 17 (2.65 g, 3.01
mmol) as described in general procedure gave 18 as a
foamy solid (3.8 g, 92%); H NMR: l 0.10, 0.11 (2 s, 6
1
1
0.08, 0.10 (2 s, 6 H, Si(CH3)2), 0.81 (t, 3 H, J 7.2 Hz,
CH2CH3), 0.92 (s, 9 H, t-Bu), 0.88–1.30 (m, 12 H, 6
CH2), 1.78, 1.84, 1.92, 2.02 (4 s, 12 H, COCH3),
3.12–3.15 (m, 1 H, OCHaHb), 3.43–3.46 (m, 1 H,
OCHaHb), 3.63 (dd, 1 H, J 10.8, 6.8 Hz, H-6a), 3.66–
3.84 (m, 4 H, H-5, H-5%, H-6a%, H-6b), 3.90 (dd, 1 H, J
10.8, 3.2 Hz, H-6b%), 4.16 (dd, 1 H, J 10.8, 8.4 Hz, H-2),
4.29 (dd, 1 H, J 10.8, 8.4 Hz, H-2%), 4.88 (t, 1 H, J 9.2
Hz, H-4), 5.14 (t, 1 H, J 9.2 Hz, H-4%), 5.17 (d, 1 H, J
8.4 Hz, H-1), 5.44 (d, 1 H, J 8.4 Hz, H-1%), 5.67 (dd, 1
H, J 10.8, 9.0 Hz, H-3), 5.77 (dd, 1 H, J 10.8, 9.2 Hz,
H-3%), 7.70–7.85 (m, 8 H, Ph); 13C NMR: l −5.36,
13.96, 18.26, 20.34, 20.40, 20.42, 20.65, 22.50, 25.71,
25.79, 29.00, 29.02, 29.06, 31.55, 54.59 (C-2%), 54.62
(C-2), 62.29 (C-6), 68.07 (C-6%), 69.35 (OCH2), 69.35
(C-4%), 69.72 (C-4), 71.70 (C-3), 71.07 (C-3%), 73.03
(C-5%), 74.66 (C-5), 97.59, 97.66 (C-1%, C-1), 123.40,
123.44, 131.37, 131.49, 134.08, 167.25, 167.56, 167.70,
167.80, 169.29, 170.02, 170.19. Anal. Calcd for
C50H66N2O17Si: C, 60.35; H, 6.68. Found: C, 60.56; H,
6.61.
H), 0.81 (t, 3 H, J 7.2 Hz), 0.93 (s, 9 H), 0.85–1.28 (m,
12 H), 1.77, 1.79, 1.85, 1.91, 1.93, 2.02 (6 s, 18 H),
3.19–3.21 (m, 1 H), 3.46 (dd, 1 H, J 10.8, 6.4 Hz),
3.49–3.52 (m, 2 H), 3.66–3.84 (m, 6 H), 3.91 (d, 1 H),
4.13 (dd, 1 H, J 10.4, 8.4 Hz), 4.20 (dd, 1 H, J 10.8, 8.4
Hz), 4.30 (dd, 1 H, J 10.8, 8.4 Hz), 4.79 (t, 1 H, J 9.2
Hz), 4.92 (t, 1 H, J 9.2 Hz), 5.13 (d, 1 H, J 8.4 Hz), 5.15
(t, 1 H, J 9.2 Hz), 5.30 (d, 1 H, J 8.4 Hz), 5.45 (d, 1 H,
J 8.4 Hz), 5.62 (dd, 2 H, J 10.8, 9.0 Hz), 5.77 (dd, 1 H,
J 10.8, 9.2 Hz), 7.77–7.92 (m, 12 H); 13C NMR: l
−5.36, −5.37, 13.96, 18.28, 20.29, 20.35, 20.39, 20.46,
20.66, 20.94, 22.50, 25.73, 25.81, 29.00, 29.04, 29.08,
31.55, 54.41, 54.53, 54.57, 62.33, 67.67, 67.90, 69.39,
69.43, 69.54, 69.64, 70.65, 70.70, 71.06, 72.72, 73.11,
74.64, 97.54, 97.62, 97.71, 123.40, 123.45, 123.61,
131.38, 131.46, 134.07, 131.46, 134.29, 169.23, 169.34,
170.00, 170.06, 171.18. Compound 18 (1.77 g, 1.29
mmol) was desilylated as described in the preparation
of 9 to give 19 (1.40 g, 86.5%) as a foam: [h]D +38° (c
1, CHCl3); 1H NMR: l 0.81 (t, 3 H, J 7.2 Hz,
CH2CH3), 0.85–1.26 (m, 12 H, 6 CH2), 1.79, 1.80, 1.86,
1.92, 1.97, 2.06 (6 s, 18 H, COCH3), 3.20–3.22 (m, 1 H,
OCHaHb), 3.48–3.51 (m, 1 H, OCHaHb), 3.53 (dd, 1 H,
J 10.8, 6.4 Hz, H-6a), 3.60–3.63 (m, 1 H, H-5), 3.66
(dd, 1 H, J 10.8, 6.4 Hz, H-6a¦), 3.71–3.85 (m, 5 H,
H-5¦, H-5%, H-6a%, H-6b, H-6b¦), 3.95–3.97 (m, 1 H,
H-6b%), 4.15 (dd, 1 H, J 10.4, 8.4 Hz, H-2), 4.23 (dd, 1
H, J 10.4, 8.4 Hz, H-2%), 4.31 (dd, 1 H, J 10.0, 8.4 Hz,
H-2¦), 4.84 (t, 1 H, J 9.2 Hz, H-4), 5.03 (t, 1 H, J 9.2
Hz, H-4%), 5.15 (t, 1 H, J 10.0 Hz, H-4¦), 5.16 (d, 1 H,
J 8.4 Hz, H-1), 5.35 (d, 1 H, J 8.4 Hz, H-1%), 5.51 (d, 1
H, J 8.4 Hz, H-1¦), 5.65 (dd, 2 H, J 10.4, 9.2 Hz, H-3,
H-3%), 5.80 (t, 1 H, J 10.0 Hz, H-3¦), 7.71–7.90 (m, 12
H, Ph); 13C NMR: l 13.91, 20.25, 20.30, 20.30, 20.47,
20.50, 20.54, 22.43, 25.66, 28.95, 28.97, 29.02, 31.49,
54.36 (C-2¦, C-2%), 54.55 (C-2), 61.15 (C-6¦), 67.92
(C-6%), 68.35 (C-6), 69.16 (C-4¦), 69.44 (OCH2), 69.85
(C-4%), 69.87 (C-4), 70.56 (C-3, C-3%), 70.75 (C-3¦), 72.51
(C-5%), 72.69 (C-5), 74.22 (C-5¦), 97.48 (C-1¦), 97.61
(C-1), 97.72 (C-1%), 123.38, 123.42, 123.55, 131.29,
131.34, 131.46, 134.08, 134.22, 169.54, 169.58, 169.88,
169.93, 169.96, 169.96. MALDITOF-MS: Calcd for
3.14. Octyl 3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-
glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-
D-
phthalimido-b-D-glucopyranoside (17)
Compound 16 (2.2 g, 2.21 mmol) was desilylated as
described in the preparation of 9 to give 17 (1.53 g,
1
78%) as a syrup: [h]D +37° (c 1, CHCl3); H NMR: l
0.81 (t, 3 H, J 7.2 Hz, CH2CH3), 0.88–1.28 (m, 12 H,
6 CH2), 1.82, 1.86, 1.98, 2.06 (4 s, 12 H, Ac), 3.20–3.22
(m, 1 H, OCHaHb), 3.51–3.53 (m, 1 H, OCHaHb),
3.64–3.73 (m, 4 H, H-5, H-5%, H-6a%, H-6a), 3.83 (dd, 1
H, J 12.0, 3.2 Hz, H-6b%), 3.90–3.92 (m, 1 H, H-6b),
4.18 (dd, 1 H, J 10.8, 8.4 Hz, H-2), 4.29 (dd, 1 H, J
10.8, 8.8 Hz, H-2%), 4.97 (t, 1 H, J 9.6 Hz, H-4), 5.14 (t,
1 H, J 9.4 Hz, H-4%), 5.20 (d, 1 H, J 8.4 Hz, H-1), 5.49
(d, 1 H, J 8.8 Hz, H-1%), 5.69 (dd, 1 H, J 10.8, 9.6 Hz,
H-3), 5.81 (dd, 1 H, J 10.8, 9.4 Hz, H-3%), 7.70–7.87 (m,
8 H, Ph); 13C NMR: l 13.96, 20.36, 20.60, 20.69, 22.50,
25.70, 29.00, 29.02, 29.09, 31.55, 54.48 (C-2%), 54.62
(C-2), 62.20 (C-6), 68.46 (C-6%), 69.23 (C-4%), 69.64
(OCH2), 70.26 (C-4), 70.64 (C-3), 70.68 (C-3%), 72.62

F. Yang, Y. Du / Carbohydrate Research 338 (2003) 495–502
501
C62H69N3O25: 1255.4 [M]; Found 1278.9 [M+Na]+.
Anal. Calcd for C62H69N3O25: C, 59.28; H, 5.54.
Found: C, 59.02; H, 5.63.
69.99, 70.60, 70.70, 70.75, 70.90 (4 C-3), 72.58, 72.77,
72.87 (C-5III, C-5II, C-5IV), 74.34 (C-5I), 97.58 (C-1I),
97.61 (C-1II), 97.69 (C-1III), 97.74 (C-1IV), 169.58,
169.65, 169.73, 170.73, 170.01, 170.04, 170.06, 170.12,
170.15. MALDITOF-MS: Calcd for C80H86N4O33:
1630.5 [M]; Found 1654.0 [M+Na]+. Anal. Calcd for
C80H86N4O33: C, 58.89; H, 5.31. Found: C, 58.68; H,
5.39.
3.16. Octyl 3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-
glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-
D-
phthalimido-b-D-glucopyranosyl-(1“6)-3,4-di-O-acetyl-
2-deoxy-2-phthalimido-b-
D-glucopyranosyl-(1“6)-3,4-
di-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyranoside
(21)
3.17. Octyl 3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-
glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-
D-
Coupling of 7 (1.212 g, 2.14 mmol) and 19 (2.56 g, 2.04
mmol) as described in general procedure gave 20 as a
foamy solid (2.403 g, 67.5%); H NMR: l 0.09, 0.11 (2
phthalimido-b-
2-deoxy-2-phthalimido-b-
di-O-acetyl-2-deoxy-2-phthalimido-b-
(1“6)-3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-
glucopyranosyl-(1“6)-3,4-di-O-acetyl-2-deoxy-2-
phthalimido-b- -glucopyranosyl-(1“6)-3,4-di-O-acetyl-
2-deoxy-2-phthalimido-b- -glucopyranosyl-(1“6)-3,4-
di-O-acetyl-2-deoxy-2-phthalimido-b- -glucopyranosyl-
D
-glucopyranosyl-(1“6)-3,4-di-O-acetyl-
-glucopyranosyl-(1“6)-3,4-
-glucopyranosyl-
D
1
D
s, 6 H, 2 SiCH3), 0.80 (t, 3 H, J 7.2 Hz, CH2CH3),
0.83–1.28 (m, 12 H, 6 CH2), 1.76, 1.78, 1.80, 1.85, 1.92,
1.93, 1.95, 2.05 (8 s, 24 H, COCH3), 3.16 (m, 1 H,
OCHaHb), 3.35–3.46 (m, 4 H, H-6aII, H-6aIII, H-5III,
OCHaHb), 3.57–3.59 (m, 1 H, H-5II), 3.61–3.65 (m, 2
H, H-6bII, H-6III), 3.73–3.85 (m, 5 H, H-5I, H-5IV,
H-6aI, H-6aIV, H-6bIV), 3.93 (dd, 1 H, J 11.2, 2.4 Hz,
H-6bI), 4.11–4.19 (m, 3 H, H-2II, H-2III, H-2I), 4.30
(dd, 1 H, J 11.6, 8.4 Hz, H-2IV), 4.75 (t, 1 H, J 9.2 Hz,
H-4III), 4.82 (t, 1 H, J 9.2 Hz, H-4II), 4.90 (t, 1 H, J 9.2
Hz, H-4I), 5.14 (d, 1 H, J 8.4 Hz, H-1III), 5.16 (t, 1 H,
J 9.2 Hz, H-4IV), 5.23 (d, 1 H, J 8.4 Hz, H-1II), 5.29 (d,
1 H, J 8.4 Hz, H-1I), 5.43 (d, 1 H, J 8.4 Hz, H-1IV), 5.56
(dd, 1 H, J 11.6, 9.2 Hz, H-3III), 5.64 (dd, 1 H, J 11.6,
9.2 Hz, H-3II), 5.65 (dd, 1 H, J 11.6, 9.2 Hz, H-3I), 5.79
(dd, 1 H, J 11.6, 9.2 Hz, H-3IV), 7.70–7.79 (m, 16 H,
Ph); Selected 13C NMR: l −5.36, 13.95, 18.27, 22.49,
25.72, 25.80, 28.99, 29.02, 29.03, 31.53, 54.35, 54.40,
54.54, 54.60 (4 C-2), 62.28 (C-6IV), 67.26 (C-6III), 67.93
(C-6I, C-6II), 69.32, 69.40, 69.50, 69.57, 69.58 (5C),
70.57, 70.61, 70.69, 71.04 (4 C-3), 72.64 (C-5III), 72.69
(C-5II), 72.95 (C-5IV), 74.58 (C-5I), 97.48 (C-1I), 97.60
(C-1II), 97.61 (C-1III), 97.74 (C-1IV), 169.22, 169.30,
169.37, 169.45, 169.95, 169.99, 170.05, 170.16. Com-
pound 20 (480 mg, 0.275 mmol) was desilylated as
described in the preparation of 9 to give 21 (377 mg,
D-
D
D
D
(1“6)-3,4-di-O-acetyl-2-deoxy-2-phthalimido-b-D-
glucopyranoside (24)
Coupling of 15 (528 mg, 0.264 mmol) and 21 (572 mg,
0.352 mmol) at −10 °C as described in general proce-
dure gave 24 as a foamy solid (443 mg, 47.4%): [h]D
1
+41° (c 1, CHCl3); H NMR: l 0.81 (t, 3 H, J 7.6 Hz,
CH2CH3), 0.85–1.28 (m, 12 H, 6 CH2), 1.77–2.16 (m,
57 H, 19 COCH3), 3.14–3.16 (m, 1 H, OCHaHb),
3.35–3.60 (m, 16 H), 3.61–3.80 (m, 8 H), 3.87–3.95 (m,
2 H), 3.41–3.43 (m, 9 H), 4.37–4.44 (m, 2 H), 4.74–
4.83 (dd, 7 H), 4.93 (t, 1 H, J 9.2 Hz), 5.12 (d, 1 H, J1,2
8.4 Hz, H-1), 5.21 (t, 1 H, J 9.2 Hz), 5.22–5.26 (m, 6 H,
6 H-1), 5.31 (d, 1 H, J 8.4 Hz, H-1), 5.50 (d, 1 H, J 8.4
Hz, H-1), 5.58–5.66 (m, 8 H), 5.80 (dd, 1 H, J 10.6, 9.2
Hz), 7.71–7.92 (m, 36 H, Ph); Selected 13C NMR: l
13.96, 20.31, 22.49, 25.72, 28.99, 29.02, 29.03, 31.53,
54.37, 54.58, 61.94, 67.18, 67.37, 67.47, 67.48, 67.57,
67.76, 67.92, 68.15, 68.13, 68.90, 69.32, 69.44, 69.62,
70.50, 70.63, 70.73, 71.98, 72.56, 72.93, 97.51, 97.58,
97.59, 97.63, 97.68, 98.01, 169.36, 169.43, 169.46,
169.89, 170.03, 171.06. MALDITOF-MS: Calcd for
1
84%) as a foam: [h]D +40° (c 1, CHCl3); H NMR: l
0.81 (t, 3 H, J 7.2 Hz, CH2CH3), 0.83–1.26 (m, 12 H,
6 CH2), 1.78, 1.79, 1.80, 1.87, 1.91, 1.95, 1.97, 2.06 (8 s,
24 H, COCH3), 3.05 (br s, 1 H, OH), 3.17–3.19 (m, 1
H, OCHaHb), 3.45–3.85 (m, 12 H), 3.95 (m, 1 H,
H-6bI), 4.11–4.24 (m, 3 H, H-2II, H-2III, H-2I), 4.33
(dd, 1 H, J 11.6, 8.4 Hz, H-2IV), 4.82 (t, 1 H, J 9.2 Hz,
H-4III), 4.85 (t, 1 H, J 9.2 Hz, H-4II), 5.03 (t, 1 H, J 9.2
Hz, H-4I), 5.14 (d, 1 H, J 8.4 Hz, H-1III), 5.17 (t, 1 H,
J 9.2 Hz, H-4IV), 5.29 (d, 1 H, J 8.4 Hz, H-1II), 5.36 (d,
1 H, J 8.4 Hz, H-1I), 5.52 (d, 1 H, J 8.4 Hz, H-1IV), 5.58
(dd, 3 H, J 11.6, 9.2 Hz, 3 H-3), 5.82 (dd, 1 H, J 11.6,
9.2 Hz, H-3IV), 7.71–7.93 (m, 16 H, Ph). Selected 13C
NMR: l 14.04, 22.58, 25.80, 29.08, 29.10, 29.12, 31.62,
54.40, 54.42, 54.51, 54.69 (4 C-2), 61.24 (C-6IV), 67.99
(C-6I, C-6II), 68.08 (C-6III), 69.31, 69.48, 69.74, 69.95,
C
172H173N9O74: 3548 [M]; Found 3572.2 [M+Na]+.
3.18. Octyl 2-amino-2-deoxy-b-
6)-2-amino-2-deoxy-b- -glucopyranosyl-(1“6)-2-amino-
2-deoxy-b- -glucopyranosyl-(1“6)-2-amino-2-deoxy-b-
-glucopyranosyl-(1“6)-2-amino-2-deoxy-b- -gluco-
pyranosyl-(1“6)-2-amino-2-deoxy-b- -glucopyranosyl-
(1“6)-2-amino-2-deoxy-b- -glucopyranosyl-(1“6)-2-
amino-2-deoxy-b- -glucopyranosyl-(1“6)-2-amino-2-
deoxy-b- -glucopyranoside (25)
D-glucopyranosyl-(1“
D
D
D
D
D
D
D
D
To NH3-satd MeOH (150 mL) was added 24 (350 mg,
0.099 mmol). The mixture was stirred at rt for 9 days,
then concentrated. The residue was dissolved in H2O (1
mL) and then passed through a Bio-Gel P-2 column to

502
F. Yang, Y. Du / Carbohydrate Research 338 (2003) 495–502
(b) Zhang, H.; Du, Y.; Yu, X.; Mitsutoni, M.; Aiba, S.
Carbohydr. Res. 1999, 320, 257–260;
(c) In Chitin in Nature and Technology; Muzzarelli, R.;
Jeuniaux, C.; Gooday, G. W., Eds.; New York: Plenum
Press, 1986.
give 25 (112 mg, 72%) as a white solid: [h]D −21° (c
0.5, CHCl3); 13C NMR (100 MHz, D2O): l 102.96,
103.08, 103.54 (3 C), 103.60 (2 C), 103.66, 103.72 (9
C-1). MALDITOF-MS: Calcd for C62H117N9O37:
1579.8 [M]; Found 1603.2 [M+Na]+.
3. Hinou, H.; Umino, A.; Matsuoka, K.; Terunuma, D.;
Takahashi, S.; Esumi, Y.; Kuzuhara, H. Tetrahedron
Lett. 1997, 38, 8041–8044.
4. Defaye J.; Gadelle A.; Pedersen C.; Fr. Demande FR
2,640,628 (1990); Chem. Abstr., 1991, 114, 247674x.
5. (a) Fridman, M.; Solomon, D.; Yogev, S.; Baasov, T.
Org. Lett. 2002, 4, 281–283;
(b) Solomon, D.; Fridman, M.; Zhang, J.; Baasov, T.
Org. Lett. 2001, 3, 4311–4314.
6. (a) Yang, F.; He, H.; Du, Y. Tetrahedron Lett., 2002, 43,
7561–7563;
Acknowledgements
This work was supported by NNSF of China
(Projects 29972053, 39970179), Beijing YFAX Sci-Tech
Ltd and RCEES of CAS. The authors thank Professor
Qi-Xiang Lu of National Institute for the Control of
Pharmaceutical and Biological Products for performing
the mice tests.
(b) Du, Y.; Yang, F. Chinese Pat. Appl. 01136484x, 2001.
7. Du, Y.; Zhang, M.; Kong, F. Tetrahedron 2001, 57,
1757–1763.
8. (a) Lemieux, R. U.; Takeda, T.; Chung, B. Y. ACS
Symp. Ser. 1976, 39, 90–115;
(b) Nakano, T.; Ito, Y.; Ogawa, T. Carbohydr. Res. 1993,
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