Synthesis of the trisaccharide portion of soyasaponin βg: Evaluation of a new glucuronic acid acceptor

Article abstract of DOI:10.1016/S0008-6215(03)00195-2

The synthesis of the trisaccharide portion of soyasaponin βg was successfully achieved using a new glucuronic acid acceptor: methyl 1-O-allyl-3,4-di-O-methoxymethyl-β-D-glucuronate (9). This compound and methyl 1-O-allyl-3,4-di-O-tert-butyldimethylsilyl-β

Full text of DOI:10.1016/S0008-6215(03)00195-2

Carbohydrate Research 338 (2003) 1441ꢁ1454
/
www.elsevier.com/locate/carres
Synthesis of the trisaccharide portion of soyasaponin bg: evaluation
of a new glucuronic acid acceptor
Karen Ple´*
Laboratoire de Pharmacognosie, Universite´ de Reims Champagne-Ardenne, CNRS UMR 6013, CPCBAI, Baˆt. 18, Moulin de la Housse, BP 1039, F-
51687 Reims, France
Received 5 March 2003; accepted 24 April 2003
Abstract
The synthesis of the trisaccharide portion of soyasaponin bg was successfully achieved using a new glucuronic acid acceptor:
methyl 1-O-allyl-3,4-di-O-methoxymethyl-b-
silyl-b-
successfully coupled to ethyl 2-O-benzoyl-3,4,6-tri-O-benzyl-1-thio-b-
D-glucuronate (9). This compound and methyl 1-O-allyl-3,4-di-O-tert-butyldimethyl-
D
-glucuronate (8) were both prepared from glucuronolactone via a glycal intermediate. The former compound 9 was
D
-galactopyranoside (13) in excellent yield. Synthesis of the
protected trisaccharide was then completed by the addition of a suitably protected rhamnose derivative to the disaccharide portion.
The reactivity of the glucuronic acid derivative 9 was also explored with trichloroacetimidate and fluoride donors.
# 2003 Elsevier Science Ltd. All rights reserved.
Keywords: DDMP saponins; Glucuronic acid glycosylation; Saponin synthesis
1. Introduction
the instability of this compound and the tedious
extraction process required to obtain even a small
quantity from natural sources. Oxygen and radical
scavenging activity have been reported,7 as well as the
antimutagenic activity of saponin extracts containing a
mixture of DDMP and non-DDMP saponins.8
Soyasaponin bg (1), a triterpenoid saponin, was first
isolated and identified by our laboratory1 as well as
others2 in 1992. Also known as Soysaponin VI1 and
Chromosaponin I,2b it is one of the major soybean
saponins, and is found in a variety of other leguminous
plants. In general, saponins are composed of oligosac-
charides of various lengths which are attached to a
triterpene or steroid aglycone. Soyasaponin bg (Fig. 1) is
unique in that it possesses a 2,3-dihydro-2,5-dihydroxy-
6-methyl-4H-pyran-4-one or ‘DDMP’ fragment at-
tached through a glycoside bond to the C-22 hydroxyl
group of the sapogenin Soyasapogenol B.
Numerous biological activities have been attributed to
group B soybean saponins,3 in particular to soyasaponin
I, the non-DDMP counterpart of soyasaponin bg. Some
of these activities include antiviral activity against HIV
in vitro,4 hepatoprotective effects,5 and sialyltransferase
inhibition.6 In comparison, the biological activity of
soyasaponin bg has been less studied, most likely due to
Chemical synthesis offers an alternative to the extrac-
tion of low abundance saponins in natural products.
The trisaccharide moiety of soyasaponin bg is composed
of a b-
a b- -galactose residue, which in turn is coupled in
position 2 to an a- -rhamnose. This trisaccharide unit,
D-glucuronic acid residue coupled in position 2 to
D
L
also called ‘fabatriose’ has been shown to be essential to
the hepatoprotective activity of certain b-fabatriosyl
oleanene glycosides from Fabaceae plants.5a
The presence of a b-
D-glucuronic acid residue coupled
in position 2 to another sugar is common in a large
variety of natural products, especially saponins. Un-
fortunately, the use of glucuronic acid derivatives as
acceptors in glycosylation reactions remains proble-
matic because the synthesis of the desired derivatives is
not always straightforward, and the reactivity of the
resulting hydroxy compound is often low. In many
cases, glucose is used as a glucuronic acid equivalent,9
and oxidation is performed at a later stage in the
* Tel.: ꢀ
/
33-3-26918419; fax: ꢀ
/
33-3-26913596.
E-mail address: karen.ple@univ-reims.fr (K. Ple´).
0008-6215/03/$ - see front matter # 2003 Elsevier Science Ltd. All rights reserved.
doi:10.1016/S0008-6215(03)00195-2

1442
K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ1454
/
tion with sodium methoxide followed by protection with
tert-butyldimethylsilyl (TBS) or methoxymethyl
(MOM) groups led to compounds 4 and 5. Treatment
of the glycals with dimethoxydioxirane15 led to the
formation of only one epoxide in the case of the TBS
derivative 6, and a mixture of epoxides when starting
from the MOM compound 5 (Scheme 1).
The orientation of the epoxide (a or b) could not be
determined in an absolute manner based solely on H
1
NMR coupling constants, but ring opening of the TBS
derivative by allyl alcohol gave the b anomer exclusively
(8), indicating that the precursor was the a epoxide.
Similar treatment of the MOM epoxide mixture resulted
in the formation of the two easily separated anomers in
an a/b ratio of 1:9, again showing that the major
epoxide was a.
Fig. 1. Soyasaponin bg.
synthesis. This, however, involves additional protection
and deprotection steps for the hydroxyl group in
position 6 as well as final oxidation to the acid. To
avoid these extra steps, we opted for the use of a
glucuronic acid derivative in spite of the potential
drawbacks.
The thioglycoside ethyl 2-O-acetyl-3,4,6-tri-O-benzyl-
1-thio-b-
D
-galactopyranoside16 10 (Scheme 2) was cho-
sen as a suitable galactose donor. The acetate group in
position 2 could be selectively deprotected after dis-
accharide formation, and would also serve to promote
b-glycosyeudation. Unfortunately, coupling of the TBS
acceptor 8 and the thioglycoside donor 10 proved
unsuccessful despite the different reaction conditions
exemplified in Scheme 2.
The preparation and use of synthetic protected
glucuronic acid derivatives having a free hydroxyl group
in position 2 has been limited. Saito and co-workers
reported the use of a 2-O-trichloroacetyl glucuronic acid
chloride obtained by treatment of methyl 2,3,4-tri-O-
One of the only identifiable products obtained in trace
amounts was the coupling product between two galac-
tose residues 11 as a result of acetate migration. The
limited reactivity of the acceptor seemed to be the origin
of the failed glycosylations for steric and/or electronic
reasons. The first positive results were obtained by using
the MOM glucuronic acceptor 9 with the thioglycoside
10 in the presence of trifluoromethanesulfonic acid
(TfOH) and N-iodosuccinimide (NIS) (Scheme 3). The
disaccharide 14 was synthesized in 50% yield, which
tended to confirm the hypothesis of steric hindrance
influencing the reactivity of the acceptor.
acetyl-b-
D
-glucuronate with PCl5 in toluene.10 In the
preparation of glycyrrhetinic acid a and b glycosides11
and sarsasapogenin glycosides,10 the glucuronic acid
derivatives were first coupled to the genine, followed by
selective deprotection of the trichloroacetyl group. The
poor yields obtained for the synthesis of the glucuronic
acid portion as well as for the glycosylation steps
discouraged the use of this method.
More recently, glucuronic acid glycals were reported
as being a possible source of 2-hydroxy glucuronic acid
derivatives through oxidation of the glycal followed by
ring opening. 2-Hydroxythioglycosides of methyl glu-
curonate have been described using dimethoxydioxirane
as the oxidizing agent.12 Gin and Kim reported the
sulfonium mediated oxidation of glucuronic acid glycals
followed by nucleophilic epoxide opening, and the use of
this sugar in their synthesis of the immunologic adjuvant
QS-21A.13 We wish to report the preparation of a new
glucuronic acid derivative and its use in the synthesis of
the trisaccharide portion of Soyasapogenin bg. The
reactivity of this compound as an acceptor in glycosyla-
tion reactions with various donors was also studied.
A series of reactions were then carried out to optimize
disaccharide formation. Although no orthoacetate for-
mation had been detected in the glycosylation reaction,
the acetate group in position 2 was replaced with a
2. Results and discussion
The starting point of the synthesis was methyl 3,4-di-O-
acetyl-D-glucuronate glycal (2), readily available from
commercially available glucuronolactone.14 Deacetyla-
Scheme 1.

K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ
/1454
1443
derivative was therefore used for the rest of the
synthesis.
The next step was the deprotection of the benzoate
ester in position 2 of the galactose residue (Scheme 5).
Deprotection under strongly basic conditions resulted in
total elimination of the protected hydroxyl group in
position 4 of the glucuronic acid portion to give 17a
which went on to lose the benzoate ester giving 17b. Use
of milder aqueous conditions resulted in the initial
saponification of the glucuronic ester, thus effectively
avoiding the elimination reaction, followed by removal
of the benzoate ester. Treatment of the crude reaction
mixture with diazomethane gave the hydroxy disacchar-
ide 18 in good yield.
Scheme 2.
Finally, coupling of the disaccharide 18 with 2,3,4-tri-
L
-rhamnopyranosyl trichloroacetimidate17
O-acetyl-a-
(19) in the presence of trimethylsilyl trifluoromethane-
sulfonate (TMSOTf) gave the desired trisaccharide in
44% yield. A further improvement was made by using
2,3,4-tri-O-benzoyl-a-L-rhamnopyranosyl trichloroace-
timidate18 (20) under the same reaction conditions
which gave 80% of the trisaccharide 22 (Scheme 5).
This rhamnose derivative also proved to be more stable
and give a cleaner reaction than the acetylated one.
Having the glucuronic acceptor 9 in hand, it became
very interesting to investigate its reactivity in the
presence of different donors. Three types of glycosyl
donors are routinely used in carbohydrate synthesis:
thioglycosides,19 trichloroacetimidates,20 and glycosyl
fluorides.21,22 Having successfully used thioglycoside
donors, trichloroacetimidates were then studied. 2-O-
Benzoyl-3,4,6-tri-O-benzyl-a-
chloroacetimidate (28a) was synthesized from
1,2,3,4,6-penta-O-acetyl- -galactose via the allyl
D
-galactopyranosyl
tri-
Scheme 3.
D
orthoester 24 (Scheme 6). One pot deacetylation/benzy-
lation of 24 followed by orthoester opening with
TMSOTf gave 26 in good overall yield. The acetate at
position 2 was then replaced with a benzoate ester 27,
and the allyl anomeric protecting group was removed
with rhodium catalysis. DBU catalysis of trichloroace-
timidate formation gave a 1:1 mixture of anomers 28a/
28b which were easily separated by column chromato-
graphy.
benzoate ester in an attempt to increase the yield of the
reaction. Glycosylation with the galactose derivative 13
at 0 8C gave a 50% yield of disaccharide 15 and lowering
the temperature to ꢂ78 8C gave 92% of the disacchar-
/
ide. When these optimized conditions were applied to
the coupling reaction between the thioglycoside 13 and
the TBS acceptor 8, disaccharide formation was ob-
served for the first time (Scheme 4), but the yields were
lower compared to glycosylation with 9. The MOM
Limited disaccharide formation was observed with the
trichloroacetimidate 28a and the glucuronic derivative 9
in the presence of TMSOTf at ꢂ
/
20 or ꢂ78 8C. The
/
major product of the reaction was identified as being the
amide 29, resulting from acid catalyzed rearrange-
ment.23 (Scheme 7). To our surprise, reaction of the b
anomer under identical conditions gave different results
as the disaccharide became the major product.
In an attempt to understand these results, two other
galactose donors were prepared in order to compare the
reactivity of the two anomers and to see if the ester at
position 2 could influence the reaction mechanism in
favor of increased disaccharide formation. The benzoate
Scheme 4.

1444
K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ1454
/
Scheme 5.
ester was replaced by a p-methoxybenzoate, and a p-
nitrobenzoate. Four new trichloroacetimidates were
thus synthesized from allyl 2-O-p-methoxybenzoyl-
3,4,6-tri-O-benzyl-
2-O-p-nitrobenzoyl-3,4,6-tri-O-benzyl-
D
-galactopyranoside (30) and allyl
-galactopyra-
D
noside (31) (see Section 3). The results of the glycosyla-
tion reaction with all of the trichloroacetimidate
derivatives and the MOM glucuronic acid acceptor 9
are summarized in Scheme 8.
The presence of a p-methoxybenzoyl in position 2 of
the galactose residue had no influence on the reaction,
but use of the b anomer gave more disaccharide at both
temperatures. Using the p-nitrobenzoate trichloroaceti-
midates changed little in the case of the a anomer, but
using the b anomer at ꢂ
/
78 8C afforded 80% of the
disaccharide.
Scheme 6.
When considering the reaction mechanism, the differ-
ence in behavior of the a and b anomers might be
explained by rapid formation of the oxonium inter-
mediate 36 (Scheme 9) in the case of the a anomer,
followed by extremely fast trichloroacetamide ‘capture’
of the oxonium ion as well as formation of the
orthoester 37 and reaction with either the acceptor
hydroxyl or the trichloroacetamide. In the case of the b
anomer, activation of the donor most likely results in
the simultaneous formation of the orthoester intermedi-
ate and trichloroacetamide departure without passing
through the oxonium intermediate, thus resulting in a
smaller amount of rearrangement product.
The net increase in yield with a p-nitrobenzoyl
protecting group in position 2 could result from an
increase in the reactivity of the intermediate orthoester
caused by the destabilizing effect of the electronegative
p-nitro group. We observed that the glycosylation
Scheme 7.
reaction at ꢂ78 8C was over within minutes using the
/

K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ
/1454
1445
Scheme 8.
p-nitrobenzoate derivative as opposed to several hours
In summary, the synthesis of the protected trisacchar-
ide 22 was completed with an overall yield of 59% from
the protected monosaccharides, giving a versatile carbo-
hydrate intermediate for the synthesis of soyasaponin bg
as well as other glycoconjugates. A novel glucuronic
acid derivative 9 was synthesized and tested in several
types of glycosylation reactions giving disaccharides in
good to excellent yields. For this type of acceptor having
a limited reactivity, thioglycosides were shown to be the
best donors followed by trichloroacetimidates, then
for the benzoate and p-methoxybenzoate ester deriva-
tives.
The use of the glycosyl fluoride donor, 2-O-benzoyl-
3,4,6-tri-O-benzyl-b-
D
-galactopyranosyl fluoride22 (38)
failed under trifluromethanesulfonic acid catalysis.22
Catalysis with silver trifluoromethanesulfonate and
hafnocene dichloride at ꢂ
/
10 and ꢂ78 8C gave the
/
desired disaccharide 15 in only 24 and 17% yield,
respectively (Scheme 10).
Scheme 9.

1446
K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ1454
/
washed with satd aq NaHCO3. The aq layer was further
extracted with EtOAc, and the combined extracts were
washed with brine, dried (Na2SO4), and concentrated.
The residue was chromatographed (1:1 cyclohexaneꢁ
/
EtOAc) to give 5 as a colorless syrup (0.555 g, 69%),
as well as 0.044 g of the mono 3-OH protected derivative
1
which was recycled; [a]D
ꢀ
/
7.68 (c 1, CHCl3); H NMR
(CDCl3): d 6.65 (d, 1 H, J1,2 6.5 Hz, H-1), 5.01 (ddd, 1
H, J2,3 5.5, J2,4 1.7 Hz, H-2), 4.83 (dd, 1 H, J4,5 2.6, J3,5
1.8 Hz, H-5), 4.8 (s, 2 H, ꢀ
/
OCH2Oꢀ
/
), 4.65 (d, 1 H, ꢀ
/
OCH2Oꢀ), 4.59 (d, 1 H, ꢀOCH2Oꢀ
/
/
/
), 4.31 (dd, 1 H, J3,4
4.2 Hz, H-4), 3.8 (ddd, 1 H, H-3), 3.73 (s, 3 H, OCH3),
3.42 (s, 3 H, OCH3), 3.36 (s, 3 H, OCH3). 13C NMR
(CDCl3): d 168.4 (COOCH3), 145.1 (C-1), 98.3 (C-2), 96
Scheme 10.
(ꢀ
/
OCH2Oꢀ
/
), 93.9 (ꢀ
/
OCH2Oꢀ/), 73.3 (C-5), 72, (C-4),
65.2 (C-3), 55.8 (OCH3), 55.6 (OCH3), 52.1 (OCH3).
Anal. Calcd for C11H18O7: C, 50.38; H, 6.92. Found: C,
50.41; H, 6.74.
glycosyl fluorides. In the case of trichloroacetimidates,
the anomeric configuration of the reacting donor as well
as the ester protecting group at position 2 can have an
important influence on the outcome of the reaction.
Work is being continued toward completion of the
synthesis of soyasaponin bg.
3.3. Methyl 1-O-allyl-3,4-di-O-tert-butyldimethylsilyl-b-
D
-glucuronate (8)
A solution of glycal 412 (1.2 g, 3.0 mmol) in CH2Cl2 (10
mL) was added dropwise to an acetone solution of
3. Experimental
dimethoxydioxirane (Â 0.095 M, 54 mL, 5.1 mmol) at
/
0 8C, and the mixture was stirred for 2 h until an NMR
sample showed the reaction to be complete. After most
of the acetone was evaporated, the residue was dissolved
in CH2Cl2, dried (Na2SO4), and filtered. The filtrate was
then evaporated to give the crude epoxide 6. Allyl
alcohol (25 mL) was added and the reaction was stirred
for 5 days at r.t. and stopped when an NMR sample of
the reaction mixture showed no more starting material.
The solvent was evaporated, and the residue chromato-
3.1. General methods
All chemicals were reagent grade and used as supplied
unless otherwise noted. Dichloromethane (CH2Cl2) and
triethylamine were refluxed over calcium hydride and
distilled prior to use. Analytical thin-layer chromato-
graphy (TLC) was performed on E. Merck Silica Gel 60
F254 plates. Compounds were visualized by dipping in
an anisaldehyde solution in ethanol and heating. Col-
umn chromatography was performed using E. Merck
Geduran Silica Gel Si 60 (40ꢁ
were recorded on a Perkinꢁ
ESIMS were recorded with a Thermofinnigan quad-
ripolar mass spectrometer with positive ion data col-
lected automatically. NMR spectra were obtained using
a Bruker Avance DRX 500 spectrometer (500 MHz for
1H and 125 MHz for 13C). Elemental analyses were
graphed on silica gel (19:1 cyclohexaneꢁ
8 as a colorless syrup (1.2 g, 83%); [a]D
CHCl3); 1H NMR (CDCl3): d 5.92 (m, 1 H, CH ꢁ
5.3 (dd, 1 H, J 17.3, J 1.6 Hz, CHꢁCH2), 5.19 (dd, 1 H,
J 10.4, J 1.3 Hz, CHꢁCH2), 4.74 (d, 1 H, J1,2 4.5 Hz, H-
1), 4.5 (ddt, 1 H, J 12.7, J 5, J 1.4 Hz, CH2ꢀCHꢁCH2),
4.25 (dd, 1 H, J3,4 J4,5 4.5 Hz, H-4), 4.19 (d, 1 H, H-5),
4.08 (dd, 1 H, J 12.7, J 6.4 Hz, CH2ꢀCHꢁCH2), 3.81
(dd, 1 H, J2,3 J3,4 4.5 Hz, H-3), 3.75 (s, 3 H,
/
EtOAc) to give
36.58 (c 1,
CH2),
ꢂ
/
/
60 mM). Optical rotations
/
/
Elmer 241 polarimeter.
/
/
/
/
ꢃ
/
/
/
ꢃ
/
performed on a PerkinꢁElmer CHN 2400.
/
COOCH3), 3.55 (ddd, 1 H, J2, OH 7.8, H-2), 3.08 (d, 1
H, OH), 0.92 (s, 9 H, tert-butyl), 0.89 (s, 9 H, tert-
3.2. Methyl 1,5-anhydro-2-deoxy-3,4-di-O-
methoxymethyl- -arabino-hex-1-enuronate (5)
butyl), 0.19 (s, 3 H, CH3Si), 0.12 (s, 6 H, 2ꢄ
(s,
H, CH3Si); 13C NMR (CDCl3):
(COOCH3), 134.3 (CHꢁCH2), 117.1 (CHꢁ
/
CH3Si), 0.1
D
3
d
169.7
/
/CH2),
Methoxymethyl chloride (1.57 g, 19.5 mmol) was added
at 0 8C to a solution of glycal 312 (0.566 g, 3.25 mmol)
and iPr2EtN (3.36 g, 26 mmol) in anhyd CH2Cl2 (25
mL). The mixture was then stirred for 48 h at room
temperature (r.t.). The reaction mixture was evaporated
to dryness and taken up in EtOAc (50 mL) which was
100.8 (C-1), 75.9 (C-5), 72.5 (C-3), 72.2 (C-2), 72.1 (C-
4), 70.1 (CH2ꢀCHꢁCH2), 52.1 (OCH3), 25.9, 25.7 (tert-
butyl), ꢂ4.2, ꢂ4.3, ꢂ4.4 (CH3Si); ESIMS: m/z 499
(Mꢀ
Na)ꢀ; Anal. Calcd for C22H44O7Si2: C, 55.42; H,
9.3. Found: C, 55.19; H, 9.61.
/
/
/
/
/
/

K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ
/1454
1447
3.4. Methyl 1-O-allyl-3,4-di-O-methoxymethyl-b-
glucuronate (9)
D
-
(CDCl3): d 165.4, 138.6, 137.8, 137.7, 133, 130.1,
129.9, 128.5, 128.3, 128.2, 128.1, 128, 127.9, 127.7,
127.5, 83.7 (C-1), 81.1 (C-3), 77.5 (C-5), 74.4, 73.6,
72.8 (C-4), 71.7, 70.2 (C-2), 68.5 (C-6), 23.7, 14.8;
A solution of glycal 5 (2.43 g, 9.3 mmol) in CH2Cl2 (20
mL) was added dropwise to an acetone solution of
ESIMS: m/z 621 (Mꢀ
/
Na)ꢀ; Anal. Calcd for
dimethoxydioxirane (Â 0.084 M, 133 mL, 11.2 mmol)
/
C36H38O6S: C, 72.22; H, 6.40. Found: C, 72.27; H, 6.55.
at 0 8C, and the mixture was stirred for 2 h until an
NMR sample showed the reaction to be complete. After
most of the acetone was evaporated, the residue was
dissolved in CH2Cl2, dried (Na2SO4), and filtered. The
filtrate was then evaporated to give the crude epoxide 7.
Allyl alcohol (30 mL) was added and the reaction was
stirred for 5 days at r.t. and stopped when an NMR
sample of the reaction mixture showed no more starting
material. The solvent was evaporated, and the residue
3.6. Methyl (2-O-acetyl-3,4,6-tri-O-benzyl-b-
galactopyranosyl)-(102)-(allyl 3,4-di-O-methoxymethyl-
b- -glucopyranosid)uronate (14)
D-
/
D
A mixture of alcohol 9 (0.082 g, 0.24 mmol), donor 10
˚
(0.196 g, 0.37 mmol), and 4 A powdered molecular
sieves (1 g) was stirred for 2 h at r.t. in CH2Cl2 (7 mL).
The mixture was cooled to ꢂ78 8C and N-iodosuccini-
/
chromatographed on silica gel (1:1 cyclohexaneꢁ
/
mide (0.082 g, 0.37 mmol) was added followed by the
dropwise addition of a triflic acid solution (0.162 mL,
EtOAc) to give 9 as a colorless syrup (2.12 g, 68%), as
well as 0.210 g of the more polar a isomer (6.8%); [a]D
0.02 mmol, Â0.15 M solution). The reaction was slowly
/
ꢀ
34.28 (c 1, CHCl3); 1H NMR (CDCl3): d 5.95 (m, 1 H,
/
allowed to warm, and was complete within 30 min. The
reaction was quenched with triethylamine and filtered
through Celite. The filtrate was washed with NaHCO3,
10% Na2S2O3, and water. The organic layer was dried
(Na2SO4), filtered and evaporated. The crude residue
was purified by column chromatography (1.5:1 cy-
CH ꢁ
5.19 (dd, 1 H, J 10.4, J 1.1 Hz, CHꢁ
OCH2Oꢀ), 4.79 (d, 1 H, ꢀOCH2Oꢀ
OCH2Oꢀ), 4.65 (d, 1 H, ꢀOCH2Oꢀ), 4.41 (d, 1 H, J1,2
7.2 Hz, H-1), 4.40 (ddt, 1 H, J 12.8, J 5.2, J 1.5 Hz,
/
CH2), 5.35 (dd, 1 H, J 17.2, J 1.4 Hz, CHꢁ
CH2), 4.84 (d, 1 H,
), 4.78 (d, 1 H, ꢀ
/
CH2),
/
ꢀ
/
/
/
/
/
/
/
/
CH2ꢀ
/
CHꢁ
/
CH2), 4.15 (ddt, 1 H, J 12.8, J 6.5, J 1.0 Hz,
CH2), 3.89 (m, 2 H, H-4, H-5), 3.82 (s, 3 H,
clohexaneꢁ
colorless syrup; [a]D
NMR (CDCl3): d 7.31 (m, 15 H), 5.82 (m, 1 H, CH ꢁ
CH2), 5.35 (dd, 1 H, J2?,3? 10, J1?,2? 8.2 Hz, H-2?), 5.22
(dd, 1 H, J 17.2, J 1.5 Hz, CHꢁCH2), 5.09 (dd, 1 H, J
10.5, J 1.3 Hz, CHꢁCH2), 4.97 (d, 1 H, CH2Ph), 4.83 (d,
1 H, ꢀOCH2Oꢀ), 4.77 (m, 2 H, H-1?, CH2Ph), 4.67 (m, 3
H, CH2Ph, ꢀOCH2Oꢀ), 4.6 (d, 1 H, CH2Ph), 4.52 (m, 2
H, H-1, ꢀOCH2Oꢀ), 4.41 (t, 2 H, CH2Ph), 4.32 (m, 1 H,
CH2ꢀCHꢁCH2), 4.02 (m, 1 H, CH2ꢀCHꢁCH2), 3.98
/
EtOAc) to give 14 (0.099 g, 50%) as a
CH2ꢀ
/
CHꢁ
/
ꢀ
/
31.98 (c 0.96, CHCl3); 1H
COOCH3), 3.5 (m, 5 H, H-2, H-3, OCH3), 3.33 (s, 3 H,
OCH3); 13C NMR (CDCl3): d 168.5 (COOCH3), 133.4
/
(CHꢁ
(2ꢄ
(C-2), 70.3 (CH2ꢀ
/
CH2), 118.2 (CHꢁ
OCH2Oꢀ), 86.2 (C-3), 76.5 (C-4), 75 (C-5), 72.6
CH2), 56.2, 56.1 (2ꢄ OCH3),
/
CH2), 101.7 (C-1), 98.3, 98
/
ꢀ
/
/
/
/
CHꢁ
/
/
/
52.5 (COOCH3); Anal. Calcd for C14H24O9: C, 50.00;
/
/
H, 7.19. Found: C, 49.79; H, 7.37.
/
/
/
/
3.5. Ethyl 2-O-benzoyl-3,4,6-tri-O-benzyl-1-thio-b-D-
galactopyranoside (13)
/
/
/
/
(d, 1 H, J3?,4? 2.4 Hz, H-4?), 3.91 (d, 1 H, J4,5 9.2 Hz, H-
5), 3.85 (t, 1 H, J 9 Hz, H-4), 3.8 (s, 3 H, OCH3), 3.68 (t,
Benzoyl chloride (0.453 mL, 3.9 mmol) was added to a
-galacto-
1 H, H-3), 3.65ꢁ3.44 (m, 4 H, H-2, H-5?, H-6a?/6b?), 3.5
/
mixture of ethyl 3,4,6-tri-O-benzyl-1-thio-b-
D
(dd, 1 H, H-3?), 3.38 (s, 3 H, OCH3), 3.31 (s, 3 H,
OCH3); 13C NMR (CDCl3): d 169.5, 168.9, 138.4, 137.8,
137.6, 133.7, 128.4, 128.2, 128.1, 128, 127.8, 127.7,
127.5, 127.4, 117, 101.8 (C-1), 101.1 (C-1?), 98.6, 98.3,
81.4 (C-2), 80.4 (C-3?), 79.3 (C-3), 77.3 (C-4), 75 (C-5),
74.4, 73.5 (C-5?), 73.5, 72.2 (C-2?), 72.1 (C-4?), 71.7, 70.5,
pyranoside16 (1.29 g, 2.6 mmol), triethylamine (1.09 mL,
7.8 mmol), and dimethylaminopyridine (DMAP) (0.064
g, 0.52 mmol) in CH2Cl2 (40 mL). The reaction was
refluxed overnight, cooled, and quenched by the addi-
tion of MeOH (3 mL). The solvent was evaporated, the
crude residue taken up in EtOAc, and the organic layer
washed with HCl (2 N), NaHCO3, and brine. After
drying, (Na2SO4), the solvent was evaporated and the
crude residue purified by column chromatography (19:1
68.2 (C-6?), 56.2, 52.4, 21; ESIMS: m/z 833 (Mꢀ
Na)ꢀ.
/
3.7. Methyl (2-O-benzoyl-3,4,6-tri-O-benzyl-b-
galactopyranosyl)-(102)-(allyl 3,4-di-O-methoxymethyl-
b- -glucopyranosid)uronate (15)
D-
cyclohexaneꢁ
solid; mp 80 8C; [a]D
(CDCl3): d 8.1 (d, 2 H, Bzꢀ
(t, 2 H, BzꢀH), 7.45ꢁ7.2 (m, 15 H), 5.78 (t, 1 H, J1,2
/
EtOAc) to give 13 (1.37 g, 88%) as a white
/
1
ꢀ
/
23.48 (c 1, CHCl3); H NMR
D
/
H), 7.65 (t, 1 H, BzꢀH), 7.52
/
/
/
ꢃ
/
A mixture of alcohol 9 (0.150 g, 0.45 mmol), donor 13
˚
(0.4 g, 0.67 mmol), and 4 A powdered molecular sieves
(2 g) was stirred for 2 h at r.t. in CH2Cl2 (15 mL). The
J2.3 9.7 Hz, H-2), 5.09 (d, 1 H, CH2Ph), 4.71 (m, 2 H,
CH2Ph), 4.55 (m, 4 H, H-1, CH2Ph), 4.12 (d, 1 H, J3,4
2.6 Hz, H-4), 3.73 (m, 4 H, H-3, H-5, H-6a/b), 2.69 (m, 2
H, CH3CH2S), 1.8 (t, 3 H, CH3CH2S); 13C NMR
mixture was cooled to ꢂ
/
78 8C and N-iodosuccinimide
(0.150 g, 0.67 mmol) was added followed by the

1448
K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ1454
/
dropwise addition of a triflic acid solution (0.297 mL,
0.04 mmol, Â0.15 M solution). The reaction was slowly
H-1?), 4.69 (d, 1 H, CH2Ph), 4.6 (d, 1 H, CH2Ph), 4.51
(d, 1 H, CH2Ph), 4.45 (m, 3 H, CH2ꢀCHꢁCH2,
CH2Ph), 4.32 (sl, 1 H, H-5), 4.21 (dl, 1 H, J3,4 2.9 Hz,
/
/
/
allowed to warm, and was complete within 30 min. The
reaction was quenched with triethylamine and filtered
through Celite. The filtrate was washed with NaHCO3,
10% Na2S2O3, and water. The organic layer was dried
(Na2SO4), filtered and evaporated. The crude residue
was purified by column chromatography (1.5:1 cy-
H-4), 4.04 (sl, 1 H, H-4?), 3.95 (dd, 1 H, J 12.3, J 5.7 Hz,
CH2ꢀ
(m, 4 H, H-2?, H-3?, H-5?, H-6b?), 0.9 (s, 9 H, tert-butyl),
0.69 (s, 9 H, tert-butyl), 0.1 (s, 6 H, 2ꢄ CH3Si), ꢂ0.22
(s, 3 H, CH3Si), ꢂ
0.25 (s, 3 H, CH3Si); 13C NMR
/
CHꢁ
/
CH2), 3.71 (m, 5 H, H-3, H-6a?, OCH3), 3.6
/
/
/
clohexaneꢁ
colorless syrup; [a]D
(CDCl3): d 8.03 (d, 2 H, Bzꢀ
(t, 2 H, BzꢀH), 7.4ꢁ7.1 (m, 15 H), 5.84 (m, 1 H, CH ꢁ
CH2), 5.64 (dd, 1 H, J2?,3? 9.9, J1?,2? 8.2 Hz, H-2?), 5.23
(dd, 1 H, J 17.3, J 1.5 Hz, CHꢁCH2), 5.1 (dd, 1 H, J
10.5, J 1.1 Hz, CHꢁCH2), 5.02 (d, 1 H, CH2Ph), 4.89 (d,
1 H, H-1?), 4.64 (m, 4 H, CH2Ph, ꢀOCH2Oꢀ), 4.54 (m, 2
H, H-1, CH2Ph), 4.5ꢁ4.4 (m, 4 H, CH2Ph, ꢀOCH2Oꢀ),
4.32 (dd, 1 H, J 12.4, J 5.1 Hz, CH2ꢀCHꢁCH2), 4.03
(m, 2 H, H-4?, CH2ꢀCHꢁCH2), 3.85 (d, 1 H, J4,5 9.1 Hz,
/
EtOAc) to give 15 (0.357 g, 92%) as a
(CDCl3): d 170.2, 164.9, 138.9, 137.8, 137.7, 134.5,
132.8, 130.3, 129.9, 128.4, 128.2, 128.1, 128, 127.9,
127.8, 127.6, 127.1, 116.5, 100.5 (C-1?), 100 (C-1), 83
(C-2), 79.8 (C-3?), 79 (C-5), 74.3, 74.2 (C-3), 73.5, 72.7
(C-5?), 72.6 (C-4?), 72.1 (C-2?), 72 (C-4), 71.3, 70.4, 68.4
1
ꢀ
/
19.38 (c 1, CHCl3); H NMR
H), 7.6 (t, 1 H, BzꢀH), 7.47
/
/
/
/
/
/
(C-6?), 52.1, 25.6, 25.5, ꢂ
1036 (Mꢀ
Na)ꢀ; Anal. Calcd for C56H76O13Si2: C,
66.37; H, 7.56. Found: C, 66.03; H, 7.38.
/
5, ꢂ
/
5.4, ꢂ
/
5.2; ESIMS: m/z
/
/
/
/
/
/
/
/
/
/
/
H-5), 3.77 (s, 3 H, OCH3), 3.69 (m, 2 H, H-4, H-6a?), 3.6
(m, 5 H, H-2, H-3, H-3?, H-5?, H-6b?), 3.25 (s, 3 H,
OCH3), 3.19 (s, 3 H, OCH3); 13C NMR (CDCl3): d
168.9, 165.3, 138.4, 137.6, 137.5, 133.7, 132.9, 130.2,
129.7, 128.4, 128.3, 128.2, 128.1, 128, 127.9, 127.6,
127.5, 117, 101.8 (C-1), 101.2 (C-1?), 98.5, 98.2, 81.8
(C-2), 80 (C-3?), 79.7 (C-3), 77.2 (C-4), 74.9 (C-5), 74.4,
73.5 (C-5?), 73.5, 72.6 (C-2?), 72 (C-4?), 71.4, 70.6, 68.2
3.9. Methyl (3,4,6-tri-O-benzyl-b-
(102)-(allyl 3,4-di-O-methoxymethyl-b-
glucopyranosid)uronate (18)
D-galactopyranosyl)-
/
D-
A solution of KOH (5% in 1:1 EtOHꢁwater, 30 mL) was
/
added to the disaccharide 15 (0.158 g, 0.018 mmol). The
reaction was stirred for 2 h at r.t., and then heated to
50 8C for another 2 h. The reaction was carefully
acidified with HCl (2 N), extracted with EtOAc, dried
(Na2SO4), and concentrated. The crude residue was
taken up in Et2O (5 mL), and treated with diazomethane
at 0 8C. Stirring was continued for 30 min at r.t.
Evaporation of the solvent followed by column chro-
(C-6?), 56.1, 56, 52.4; ESIMS: m/z 895 (Mꢀ
Na)ꢀ;
Anal. Calcd for C48H56O15: C, 66.04; H, 6.47. Found: C,
66.35; H, 6.54.
/
3.8. Methyl (2-O-benzoyl-3,4,6-tri-O-benzyl-b-
galactopyranosyl)-(102)-(allyl 3,4-di-O-tert-
butyldimethylsilyl-b- -glucopyranosid)uronate (16)
D-
/
matography (1.5:1 cyclohexaneꢁ/EtOAc) of the crude
D
product gave 18 (0.127 g, 91%) as a colorless syrup; [a]D
1
ꢀ
/
7.98 (c 1, CHCl3); H NMR (CDCl3): d 7.45ꢁ
15 H), 5.8 (m, 1 H, CH ꢁCH2), 5.22 (dd, 1 H, J 17.3, J
1.7 Hz, CHꢁCH2), 5.09 (dd, 1 H, J 10.5, J 1.5 Hz, CHꢁ
CH2), 4.94 (d, 1 H, CH2Ph), 4.88 (m, 2 H, ꢀOCH2Oꢀ),
4.82 (d, 1 H, CH2Ph), 4.8 (d, 1 H, ꢀOCH2Oꢀ), 4.75 (d, 1
H, CH2Ph), 4.68 (d, 1 H, ꢀOCH2Oꢀ), 4.62 (d, 1 H,
/
7.2 (m,
A mixture of alcohol 8 (0.030 g, 0.06 mmol), donor 13
/
˚
(0.056 g, 0.09 mmol), and 4 A powdered molecular
sieves (400 mg) was stirred for 2 h at r.t. in CH2Cl2 (5
/
/
/
/
mL). The mixture was cooled to ꢂ78 8C and N-
/
/
/
iodosuccinimide (0.021 g, 0.09 mmol) was added fol-
lowed by the dropwise addition of a triflic acid solution
/
/
CH2Ph), 4.58 (d, 1 H, J1,2 6.6 Hz, H-1), 4.48 (d, 1 H,
(0.041 mL, 0.006 mmol, Â0.15 M solution). The
/
J1?,2? 7.7 Hz, H-1?), 4.42 (d, 1 H, CH2Ph), 4.39 (d, 1 H,
reaction was slowly allowed to warm, and was complete
within 30 min. The reaction was quenched with triethy-
lamine and filtered through Celite. The filtrate was
washed with NaHCO3, 10% Na2S2O3, and water. The
organic layer was dried (Na2SO4), filtered and evapo-
rated. The crude residue was purified by column
CH2Ph), 4.31 (m, 1 H, CH2ꢀ
CH2ꢀCHꢁCH2), 3.99 (dd, 1 H, J2?,3? 9.8, Hz, H-2?), 3.92
(d, 1 H, J3?,4? 2.5 Hz, H-4?), 3.88 (m, 2 H, H-4, H-5), 3.8
(s, 3 H, OCH3), 3.7 (m, 2 H, H-2, H-3), 3.65ꢁ3.5 (m, 3
H, H-5?, H-6a?/6b?), 3.49 (d, 1 H, H-3?), 3.47 (s, 3 H,
OCH3), 3.33 (s, 3 H, OCH3); 13C NMR (CDCl3): d
168.7, 138.6, 138.5, 137.7, 133.7, 128.4, 128.2, 128.1,
127.9, 127.8, 127.5, 127.4, 116.7, 104.7 (C-1?), 101.9 (C-
1), 98.7, 98.1, 82.4 (C-2), 82.1 (C-3), 81.5 (C-3?), 77.1 (C-
4), 74.7, 74.3 (C-5), 73.5 (C-5?), 73.4, 73.3 (C-4?), 72.7,
72.3 (C-2?), 70.5, 68.3 (C-6?), 56.4, 56.3, 52.5; Anal.
/
CHꢁ
/
CH2), 4.03 (m, 1 H,
/
/
/
chromatography (9:1 cyclohexaneꢁ
(0.043 g, 67%) as a colorless syrup; [a]D
CHCl3); 1H NMR (CDCl3): d 8.05 (d, 2 H, Bzꢀ
(t, 1 H, BzꢀH), 7.45 (t, 2 H, BzꢀH), 7.4ꢁ7.1 (m, 15 H),
5.91 (m, 1 H, CH ꢁCH2), 5.6 (dd, 1 H, J2?,3? 10, J1?,2? 8.1
Hz, H-2?), 5.28 (dl, 1 H, J 17.3 Hz, CHꢁCH2), 5.13 (m,
2 H, H-1, CHꢁCH2), 5.08 (d, 1 H, CH2Ph), 4.75 (d, 1 H,
/
EtOAc) to give 16
17.08 (c 0.94,
H), 7.59
ꢀ
/
/
/
/
/
/
/
Calcd for C41H52O14×
/0.3 H2O: C, 63.60; H, 6.85. Found:
/
C, 63.40; H, 6.59.

K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ
/1454
1449
1
3.10. Methyl (2,3,4-tri-O-acetyl-a-
L-rhamnopyranosyl)-
g, 80%) as a colorless oil; [a]D
ꢀ
/
49.68 (c 1, CHCl3); H
(10
/
2)-(3,4,6-tri-O-benzyl-b-
D
-galactopyranosyl)-(10
/
NMR (CDCl3): d 8.05 (t, 4 H, Bzꢀ
H), 7.62 (t, 1 H, BzꢀH), 7.58ꢁ7.1 (m, 23 H), 5.86 (m, 3
H, H-2ƒ, H-3ƒ, CH ꢁCH2), 5.67 (t, 1 H, J3ƒ,4ƒꢃJ4ƒ,5ƒ 10
Hz, H-4ƒ), 5.57 (bs, 1 H, H-1ƒ), 5.31 (dd, 1 H, J 17.2, J
1.5 Hz, CHꢁCH2), 5.11 (dd, 1 H, J 10.5, J 1.2 Hz, CHꢁ
CH2), 5.02 (d, 1 H, ꢀOCH2Oꢀ), 4.96 (d, 1 H, ꢀ
OCH2Oꢀ), 4.91 (d, 1 H, CH2Ph), 4.84 (m, 2 H, H-1?,
OCH2Oꢀ), 4.72 (m, 5 H, H-1, H-5ƒ, CH2Ph, ꢀ
OCH2Oꢀ ), 4.58 (d, 1 H, CH2Ph), 4.5 (d, 1 H,
CH2Ph), 4.48 (d, 1 H, CH2Ph), 4.34 (m, 1 H, CH2ꢀ
CHꢁCH2), 4.2 (t, 1 H, J3,4 J4,5 9.0 Hz, H-4), 4.15
(dd, 1 H, J1?,2?ꢃJ2?,3? 8.0 Hz, H-2?), 4.05 (m, 3 H, H-2,
H-5, CH2ꢀCHꢁCH2), 3.98 (d, 1 H, J3?,4? 2.5 Hz, H-4?),
/
H), 7.84 (d, 2 H, Bzꢀ
/
2)-(allyl 3,4-di-O-methoxymethyl-b-
D-
/
/
glucopyranosid)uronate (21)
/
/
A mixture of alcohol 18 (0.087 g, 0.11 mmol), donor
1917 (0.074 g, 0.17 mmol), and 4 A powdered molecular
sieves (350 mg) was stirred for 1 h at r.t. in CH2Cl2 (5
/
/
˚
/
/
/
/
mL). The mixture was cooled to ꢂ
/
78 8C followed by the
ꢀ
/
/
/
dropwise addition of a 0.1 M solution of TMSOTf in
CH2Cl2 (0.056 mL, 0.006 mmol, 0.05 equiv). The
reaction was slowly allowed to warm until TLC
indicated that no more rhamnose imidate was present.
The reaction was quenched with triethylamine, filtered
through Celite, and evaporated. The crude residue was
purified by column chromatography (1:1.5 cy-
clohexaneꢁ
44%) as a colorless oil; H NMR (CDCl3): d 7.3 (m, 15
H), 5.86 (m, 1 H, CH ꢁCH2), 5.38 (dd, 1 H, J2ƒ,3ƒ 3.4,
J1ƒ,2ƒ 1.6 Hz, H-2ƒ), 5.29 (m, 3 H, H-1ƒ, H-3ƒ, CHꢁ
CH2), 5.11 (dd, 1 H, J 10.5, J 1.3 Hz, CHꢁCH2), 5.07
(t, 1 H, J3ƒ,4ƒꢃJ4ƒ,5ƒ 13.5 Hz, H-4ƒ), 4.88 (m, 2 H, ꢀ
OCH2Oꢀ, CH2Ph), 4.82 (m, 2 H, ꢀOCH2Oꢀ), 4.7 (m, 4
H, H-1, H-1?, ꢀOCH2Oꢀ, CH2Ph), 4.55 (d, 2 H,
CH2Ph), 4.45 (d, 2 H, CH2Ph), 4.34 (m, 2 H, H-5ƒ,
CH2ꢀCHꢁCH2), 4.21 (t, 1 H, J3,4 J4,5 9.1 Hz, H-4),
4.02 (m, 4 H, H-2, H-2?, H-5?, CH2ꢀCHꢁCH2), 3.95 (s,
/
/
/
ꢃ
/
/
/
/
3.9 (dd, 1 H, J2,3 5.7 Hz, H-3), 3.7 (dd, 1 H, J3?,4? 2.7 Hz,
H-3?), 3.62 (m, 6 H, H-5?, H-6a?/6b?, OCH3), 3.49 (s, 3
H, OCH3), 3.33 (s, 3 H, OCH3), 1.38 (d, 3 H, J5ƒ,6ƒ 6.2
Hz, H-6ƒ); 13C NMR (CDCl3): d 169.7, 165.8, 165.4,
165.2, 138.4, 137.8, 137.3, 133.7, 133.2, 133.1, 132.9,
129.9, 129.8, 129.6, 129.5, 129.4, 128.4, 128.3, 128.2,
127.9, 127.8, 127.7, 127.5, 117.2, 100.2 (C-1), 99.7 (C-1?),
98.3 (C-1ƒ), 98.1, 97.6, 83.4 (C-3?), 80.4 (C-3), 77.5 (C-2),
76.3 (C-4), 74.7 (C-2?), 74.5, 74.3 (C-5), 73.5, 73.4 (C-5?),
72.7 (C-4?), 72.5, 71.8 (C-4ƒ), 70.4 (C-2ƒ), 70.3 (C-3ƒ),
69.3, 68.4 (C-6?), 66.7 (C-5ƒ), 56.4, 56.2, 52.1, 17.3 (C-
/
Et2O) to give the trisaccharide 21 (0.052 g,
1
/
/
/
/
/
/
/
/
/
/
/
/
ꢃ
/
6ƒ); ESIMS: m/z 1250 (Mꢀ
/
Na)ꢀ; Anal. Calcd for
/
/
C68H74O21: C, 66.55; H, 6.08. Found: C, 66.32; H, 6.09.
1 H, H-4?), 3.79 (s, 3 H, OCH3), 3.75 (dd, 1 H, J2,3 5.1
Hz, H-3), 3.59 (m, 4 H, H-3?, H-5?, H-6a?/6b?), 3.42 (s, 3
H, OCH3), 3.32 (s, 3 H, OCH3), 2.09, (s, 3 H, COCH3),
2.06 (s, 3 H, COCH3), 1.98 (s, 3 H, COCH3), 1.2 (d, 3
H, J5ƒ,6ƒ 6.2 Hz, H-6ƒ); 13C NMR (CDCl3): d 170.4,
170.2, 169.9, 169.8, 138.3, 137.6, 137.3, 133.7, 128.4,
128.2, 128, 127.9, 127.8, 127.6, 117.3, 99.9 (C-1), 99.5
(C-1?), 98.1 (C-1ƒ), 98, 97.4, 83.3 (C-3?), 80.4 (C-3), 77.6
(C-2), 76.1 (C-4), 74.7 (C-2?), 74.5 (C-5), 74.5, 73.5, 73.3
(C-5?), 72.5 (C-4?), 72.3, 71 (C-4ƒ), 69.2 (C-2ƒ, C-3ƒ),
68.4, 67.5 (C-6?), 66.3 (C-5ƒ), 56.2, 56.1, 52.3, 20.8, 20.7,
3.12. 3,4,6-Tri-O-acetyl-1,2-O-(allyloxyethylidene)-a-D-
galactopyranose (24)
A solution of HBr in AcOH (33%, 3.7 mL) was slowly
added to a stirred solution of 1,2,3,4,6-penta-O-acetyl-
D-galactopyranose (1 g, 2.6 mmol) in CH2Cl2 (7 mL) at
0 8C. After 90 min at r.t., the solvent was evaporated
and toluene (20 mL) was distilled off three times from
the residue. This was taken up in nitromethane (6.5 mL)
and, after addition of 2,6-lutidine (0.45 mL, 3.84 mmol),
allyl alcohol (0.70 mL, 10.2 mmol) and tetrabutylam-
monium bromide (0.082 g, 0.26 mmol), heated to 40 8C
for 20 h. The solution was then partitioned between
EtOAc and aq NaHCO3. The aq layer was extracted
17 (C-6ƒ); ESIMS: m/z 1064 (Mꢀ
/
Na)ꢀ.
3.11. Methyl (2,3,4-tri-O-benzoyl-a-
rhamnopyranosyl)-(102)-(3,4,6-tri-O-benzyl-b-
galactopyranosyl)-(102)-(allyl 3,4-di-O-methoxymethyl-
b- -glucopyranosid)uronate (22)
L-
/
D-
/
with EtOAc (3ꢄ
residue was purified by column chromatography (4:1 to
3:1 cyclohexaneꢁEtOAc, 1% Et3N) to give 24 as a
/
), dried (Na2SO4), and evaporated. The
D
/
A mixture of alcohol 18 (0.160 g, 0.21 mmol), donor
2018 (0.223 g, 0.37 mmol), and 4 A powdered molecular
sieves (800 mg) was stirred for 1 h at r.t. in CH2Cl2 (20
colorless oil composed of an unequal mixture of endo/
1
exo isomers (0.86 g, 86%); H NMR (CDCl3): [major
˚
isomer (a), minor isomer (b)] d 5.9 (m, 2 H, CH ꢁ
/
mL). The mixture was cooled to ꢂ
/
78 8C followed by the
CH2(a,b)), 5.82 (d, 1 H, J1,2 4.8 Hz, H-1a), 5.7 (d, 1
H, J1,2 5.2 Hz, H-1b), 5.44 (m, 3 H, H-3b, H-4b, H-4a),
dropwise addition of a 0.1 M solution of TMSOTf in
CH2Cl2 (0.104 mL, 0.01 mmol, 0.05 equiv). The reaction
was slowly allowed to warm, and was complete within 1
h. The reaction was quenched with triethylamine,
filtered through Celite, and evaporated. The crude
residue was purified by column chromatography (3:1
5.32 (dd, 1 H, J 16.2, J 1.5 Hz, CHꢁ
H, J 17.2, J 1.6 Hz, CHꢁCH2(a)), 5.2 (dd, 1 H, J 10.5, J
1.4 Hz, CHꢁCH2(b)), 5.17 (dd, 1 H, J 10.5, J 1.4 Hz,
CHꢁCH2(a)), 5.07 (dd, 1 H, J3,4 6.8, J2,3 3.4 Hz, H-3a),
4.37 (ddd, 1 H, J5,6a J5,6b 6.6, J4,5 1.4 Hz, H-5b), 4.33
(m, 2 H, H-2a, H-5a), 4.23 (dd, 1 H, H-2b), 4.2ꢁ4.1 (m,
/
CH2(b)), 5.29 (dd, 1
/
/
/
ꢃ
/
cyclohexaneꢁ
/
EtOAc) to give the trisaccharide 22 (0.206
/

1450
K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ1454
/
1
6 H, H-6a/b (a), H-6a/b (b),), CH2ꢀ
/
CHꢁ
/
CH2 (a,b)),
2.05 (m, 18 H,
solid; mp 64 8C; [a]D
(CDCl3): d 7.35 (m, 15 H), 5.89 (m, 1 H, CH ꢁ
5.47 (dd, 1 H, J2,3 10.0, J1,2 8.0 Hz, H-2), 5.3 (dd, 1 H, J
17.2, J 1.5 Hz, CHꢁCH2), 5.19 (dd, 1 H, J 10.5, J 1.4
Hz, CHꢁCH2), 5.0 (d, 1 H, CH2Ph), 4.73 (d, 1 H,
ꢂ
/
5.38 (c 1, CHCl3); H NMR
4.06 (m, 2 H, CH2ꢀCHꢁCH2 (a,b)), 2.15ꢁ
/
/
/
/CH2),
CH3 (a, b)), 1.72 (s, 3 H, orthoester-CH3(a)), 1.62 (s, 3
H, orthoester-CH3(b)); 13C NMR (CDCl3): d 170.5,
170, 169.9, 169.8, 134, 133.7, 117, 116.7, 97.9, 97.4 (C-1),
73.8, 73.6 (C-2), 71.6, 71.3 (C-3), 69.1 (C-5), 66.2, 65.9
(C-4), 64.7, 64, 61.5, 61.3 (C-6), 23.8, 23.1, 20.7, 20.5;
/
/
CH2Ph), 4.65 (d, 1 H, CH2Ph), 4.58 (d, 1 H, CH2Ph),
4.52 (d, 1 H, CH2Ph), 4.48 (d, 1 H, CH2Ph), 4.45 (d, 1 H,
ESIMS: m/z 411 (Mꢀ
/
Na)ꢀ; Anal. Calcd for
H-1), 4.38 (dd, 1 H, J 13.4, J 4.7 Hz, CH2ꢀ
/
CHꢁ
/CH2),
C17H24O10: C, 52.58; H, 6.23. Found: C, 52.81; H, 6.53.
4.1 (dd, 1 H, J 13.4, J 6 Hz, CH2ꢀCHꢁCH2), 4.02 (d, 1
/
/
H, J3,4 2.5 Hz, H-4), 3.69 (m, 2 H, H-6), 3.62 (t, 1 H, J5,6
5.9 Hz, H-5), 3.58 (dd, 1 H, H-3); 13C NMR (CDCl3): d
169.5, 138.4, 138, 137.8, 133.9, 128.4, 128.2, 128.1, 128,
127.8, 127.7, 127.5, 127.4, 116.9, 100.2 (C-1), 80.3 (C-3),
74.4, 73.6 (C-5), 73.55, 72.5 (C-4), 71.9, 71.3 (C-2), 69.2,
3.13. 3,4,6-Tri-O-benzyl-1,2-O-(allyloxyethylidene)-a-D-
galactopyranose (25)
To a solution of orthoester 24 (1.03 g, 2.8 mmol) and
benzyl bromide (1.5 g, 8.9 mmol) in dry THF (7 mL)
was added powdered KOH (1.75 g, 31.2 mmol) and the
mixture was refluxed for 3 h with stirring. After the
mixture was cooled, EtOAc was added, and the solution
68.6 (C-6), 21; ESIMS: m/z 555 (Mꢀ
Na)ꢀ; Anal. Calcd
for C32H36O7: C, 72.16; H, 6.81. Found: C, 72.03; H,
6.54.
/
was successively washed with water (3ꢄ
/
20 mL), a satd
3.15. Allyl 2-O-benzoyl-3,4,6-tri-O-benzyl-b-D-
galactopyranose (27)
NaHCO3 soln (2ꢄ20 mL), and water (2ꢄ
/
/20 mL). The
organic layer was dried (Na2SO4), evaporated, and the
crude residue was purified by column chromatography
To a freshly prepared solution of NaOMe (0.1 M, 200
mL) was added the acetate 26 (5.97 g, 11.2 mmol), and
the reaction was stirred for 2 h at r.t., and then heated to
50 8C overnight. The mixture was neutralized with
Amberlite IR 120 (Hꢀ form), and concentrated to
give the deacetylated product (5.4 g, 98%), as a white
solid, pure enough for use in the next step.
Benzoyl chloride (1.35 mL, 11.7 mmol) was added to
a mixture of the above alcohol (2.86 g, 5.8 mmol),
triethylamine (3.25 mL, 23.3 mmol), and DMAP (0.142
g, 1.2 mmol) in CH2Cl2 (50 mL). The reaction was
refluxed overnight, cooled, and quenched by the addi-
tion of MeOH (3 mL). The solvent was evaporated, the
crude residue taken up in EtOAc, and the organic layer
washed with HCl (2 N), NaHCO3, and brine. After
drying, (Na2SO4), the solvent was evaporated and the
crude residue purified by column chromatography (19:1
(19:1 to 9:1 cyclohexaneꢁEtOAc, 1% Et3N) to give 25 as
/
a colorless oil composed of an unequal mixture of
isomers (endo/exo) (1.2 g, 81%); 1H NMR (CDCl3):
[major isomer (a), minor isomer (b)] d 7.4 (m, 30 H),
5.96 (m, 2 H, CH ꢁ
H-1a), 5.65 (d, 1 H, J1,2 4.5 Hz, H-1b), 5.34 (m, 2 H,
CH2(a,b)), 5.0 (d, 2 H,
/
CH2(a,b)), 5.81 (d, 1 H, J1,2 4.4 Hz,
CHꢁ
/
CH2(a,b)), 5.2 (m, 2 H, CHꢁ
/
CH2Ph), 4.88 (d, 2 H, CH2Ph), 4.74 (m, 2 H, CH2Ph),
4.68 (d, 2 H, CH2Ph), 4.54 (m, 5 H, CH2Ph, H-2a), 4.42
(dd, 1 H, J2,3 5.7 Hz, H-2b), 4.2ꢁ/4.05 (m, 8 H, H-4(a,b),
H-5(a,b), CH2ꢀCHꢁCH2 (a,b)), 4.0 (dd, 1 H, J2,3 6.0,
/
/
J3,4 2.1 Hz, H-3b), 3.69 (m, 5 H, H-3a, H-6a/b (a), H-6a/
b (b)), 1.68 (s, 3 H, orthoester-CH3(a)), 1.65 (s, 3 H,
orthoester-CH3(b)); 13C NMR (CDCl3): d 138.4, 138.3,
138.1, 138, 137.9, 137.8, 134.6, 134.4, 128.1, 127.9,
127.8, 127.7, 127.5, 122.3, 121.8, 116.6, 116.3, 97.7,
96.9 (C-1), 80.3, 80.2 (C-3), 79.7, 79.2 (C-2), 74.5, 73.5,
73.4, 73.2 (C-4), 73.1, 73 (C-5), 71.5, 71.4, 68 (C-6), 63.7,
to 4:1 cyclohexaneꢁ
white solid; mp 101 8C; [a]D
NMR (CDCl3): d 8.05 (d, 2 H, Bzꢀ
H), 7.49 7.6 (t, 2 H, BzꢀH), 7.42ꢁ7.1 (m, 15 H), 5.78 (m,
1 H, CH ꢁCH2), 5.7 (dd, 1 H, J2,3 10, J1,2 8 Hz, H-2),
5.21 (dd, 1 H, J 17.3, J 1.6 Hz, CHꢁCH2), 5.09 (dd, 1
H, J 10.5, J 1.3 Hz, CHꢁCH2), 5.02 (d, 1 H, CH2Ph),
4.68 (d, 2 H, CH2Ph), 4.58 (d, 1 H, H-1), 4.51 (m, 3 H,
CH2Ph), 4.33 (ddt, 1 H, J 13.4, J 4.7, J 1.6 Hz, CH2ꢀ
CHꢁCH2), 4.09 (ddt, 1 H, J 13.4, J 6.2 J 1.4 Hz, CH2ꢀ
CHꢁCH2), 4.04 (d, 1 H, J3,4 2.7 Hz, H-4), 3.7 (m, 4 H,
/
EtOAc) to give 27 (2.95 g, 85%) as a
1
ꢀ20.78 (c 1, CHCl3); H
/
63.5, 24.9, 24; ESIMS: m/z 555 (Mꢀ
/
Na)ꢀ; Anal. Calcd
/
H), 7.6 (t, 1 H, Bzꢀ
/
for C32H36O7: C, 72.16; H, 6.81. Found: C, 72.47; H,
7.13.
/
/
/
/
3.14. Allyl 2-O-acetyl-3,4,6-tri-O-benzyl-b-
galactopyranose (26)
D-
/
/
To a solution of orthoester 25 (6.7 g, 12.6 mmol) in
˚
CH2Cl2 (40 mL) was added 4 A molecular sieves (2 g),
and the mixture was stirred for 30 min. The solution was
cooled to 0 8C, and TMSOTf (0.251 mL, 1.38 mmol)
was added. After stirring for 2 h, the reaction was
quenched by the addition of Et3N, diluted with CH2Cl2,
/
/
/
H-3, H-5, H-6a/b); 13C NMR (CDCl3): d 165.3, 138.4,
137.8, 137.6, 133.8, 132.8, 130.2, 129.8, 128.4, 128.3,
128.2, 127.9, 127.8, 127.6, 127.5, 117, 100.1 (C-1), 79.9
(C-3), 74.4, 73.7 (C-5), 73.6, 72.3 (C-4), 71.8 (C-2), 71.1,
washed with water (2ꢄ
trated. Column chromatography (6:1 cyclohexaneꢁ
EtOAc) of the residue gave 26 (5.33 g, 77%) as a white
/), dried (Na2SO4), and concen-
69.3, 68.6 (C-6); ESIMS: m/z 617 (Mꢀ
Na)ꢀ; Anal.
Calcd for C37H38O7: C, 74.73; H, 6.44. Found: C, 74.88;
H, 6.36.
/
/

K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ
/1454
1451
3.16. 2-O-Benzoyl-3,4,6-tri-O-benzyl-a/b-
galactopyranosyl trichloroacetimidate (28)
D-
70.6 (C-2), 68 (C-6); Anal. Calcd for C36H34Cl3NO7: C,
61.86; H, 4.9; N, 2.0. Found: C, 61.90; H, 4.92; N, 1.96.
To a stirring solution of 27 (2.58 g, 4.33 mmol) in a
mixture of toluene (41 mL), EtOH (95%, 105 mL), and
water (14 mL) was added tris(triphenylphosphine)rho-
dium(I) chloride (0.681 g, 0.74 mmol) followed by 1,4-
diazabicyclo[2.2.2]octane (DABCO) (0.365 g, 3.2
mmol). The mixture was stirred at r.t. for 1 h, and
then refluxed for 17 h. The resulting brown solution was
3.17. Preparation of disaccharides using glycosyl
trichloroacetimidates
Two reactions were carried out with each glycosyl
trichloroacetimidate (a or b): coupling at ꢂ
78 8C. General procedure: a mixture of methyl 1-O-
allyl-3,4-di-O-methoxymethyl-b- -glucuronate (9) (1.0
/
20 or ꢂ
/
D
evaporated, and the residue dissolved in 9:1 acetoneꢁ
/
equiv), a or b glycosyl trichloroacetimidate (1.5 equiv),
water (150 mL). To this solution was added mercury(II)
chloride (11 g, 43.3 mmol) followed by yellow mercur-
y(II) oxide (0.047 g, 0.21 mmol), and the reaction was
stirred at r.t. for 6 h. The solvent was then evaporated,
the residue diluted with CH2Cl2, and filtered through a
pad of Celite. The pad was rinced with CH2Cl2, and the
combined filtrates were washed with 20% aq NaI and
water. The organic layer was dried (Na2SO4), evapo-
rated, and the crude residue purified by column
˚
and 4 A powdered molecular sieves was stirred for 1 h at
r.t. in CH2Cl2. The mixture was cooled and TMSOTf
(0.05 equiv, 0.1 M in CH2Cl2) was added. The reaction
was quenched with triethylamine, filtered through
Celite, and evaporated. The crude residue was purified
by column chromatography to give the disaccharide.
3.18. Methyl (2-O-benzoyl-3,4,6-tri-O-benzyl-b-
galactopyranosyl)-(102)-(allyl 3,4-di-O-methoxymethyl-
b- -glucopyranosid)uronate (15)
D-
chromatography (4:1 cyclohexaneꢁ
deprotected 2-O-benzoyl-3,4,6-tri-O-benzyl-a/b-
lactopyranoside as an oily mixture of epimers (1.98 g,
83%).
/
EtOAc) to give the
D
-ga-
/
D
To a solution of the above compound (1 g, 1.8 mmol)
in CH2Cl2 (25 mL) was added trichloroacetonitrile (1.8
mL) followed by a catalytic amount of DBU (several
drops). The reaction was stirred at r.t. for 4 h. The
solvent was then evaporated and the crude residue
purified by column chromatography (19:1 to 9:1
3.18.1. Using the a glycosyl trichloroacetimidate. Cou-
pling of the acceptor 9 (30 mg, 89 mmol) and donor 28a
(93 mg, 130 mmol) using TMSOTf (44 mL, 4 mmol)
according to the general procedure at ꢂ
gave, after purification (1.5:1 cyclohexaneꢁ
disaccharide 15 (0.016 g, 21%) as well as the rearrange-
ment product 29 (0.054 g, 68%). Reaction at ꢂ78 8C (2
/
20 8C for 20 min
/EtOAc), the
cyclohexaneꢁEtOAc, 1% Et3N) to give the a trichlor-
/
/
oacetimidate 28a as an oil (0.400 g, 32%) and the b
trichloroacetimidate 28b as a white solid (0.390 g, 31%).
h) gave 27% of the disaccharide and 69% of the
rearrangement product.
a Anomer: [a]D
(CDCl3): d 8.45 (s, 1 H, NH), 8.0 (d, 2 H, Bzꢀ
1 H, BzꢀH), 7.5ꢁ7.25 (m, 17 H), 6.69 (d, 1 H, J1,2 3.6
ꢀ
/
89.58 (c 1, CHCl3); 1H NMR
/
H), 7.6 (t,
/
/
3.18.2. Using the b glycosyl trichloroacetimidate. Cou-
pling of the acceptor 9 (25 mg, 74 mmol) and donor 28b
(78 mg, 111 mmol) using TMSOTf (37 mL, 3.7 mmol)
Hz, H-1), 5.85 (dd, 1 H, J2,3 9.9 Hz, H-2), 5.05 (d, 1 H,
CH2Ph), 4.77 (d, 1 H, CH2Ph), 4.69 (d, 1 H, CH2Ph),
4.68 (d, 1 H, CH2Ph), 4.54 (d, 1 H, CH2Ph), 4.49 (d, 1 H,
CH2Ph), 4.28 (m, 2 H, H-4, H-5), 4.22 (bs, 1 H, H-3),
3.75 (dd, 1 H, J6a,6b 9.2, J6a,5 9.0 Hz, H-6a), 3.66 (dd, 1
H, J6b,5 5.5 Hz, H-6b); 13C NMR (CDCl3): d 165.6,
according to the general procedure at ꢂ
gave, after purification (1.5:1 cyclohexaneꢁ
/
20 8C for 20 min
EtOAc), the
/
disaccharide 15 (0.043 g, 66%) identical to the above
described product as well as the rearrangement product
160.5 (Cꢁ
/
NH), 138.2, 137.7, 137.6, 133.1, 129.8, 129.6,
29 (0.033 g, 42%). Reaction at ꢂ78 8C (2 h) gave 65% of
the disaccharide and 15% of the rearrangement product.
For the rearrangement product 2-O-benzoyl-3,4,6-tri-
/
128.4, 128.3, 128.2, 128.1, 128, 127.9, 127.8, 127.7, 94.6
(C-1), 75.5 (C-3), 74.9, 73.6 (C-4), 73.5, 72.3 (C-5), 72.1,
70.1 (C-2), 68.1 (C-6).
O-benzyl-N-trichloroacetyl-b-
29: 1H NMR (CDCl3): d 7.99 (m, 2 H, Bzꢀ
2 H, NH, BzꢀH), 7.5ꢁ7.2 (m, 17 H), 5.68 (t, 1 H, J 9.6
D-galactopyranosylamine
1
b Anomer: mp 142 8C; [a]D
NMR (CDCl3): d 8.57 (s, 1 H, NH), 8.0 (d, 2 H, Bzꢀ
7.59 (t, 1 H, BzꢀH), 7.5ꢁ7.15 (m, 17 H), 5.94 (dd, 1 H,
J2,3 9.8, J1,2 8.2 Hz, H-2), 5.86 (d, 1 H, H-1), 5.05 (d, 1
H, CH2Ph), 4.7 (m, 2 H, CH2Ph), 4.5 (m, 3 H, CH2Ph),
4.11 (d, 1 H, J3,4 1.8 Hz, H-4), 3.89 (t, 1 H, J 6.1 Hz, H-
5), 3.75 (m, 3 H, H-3, H-6a/b); 13C NMR (CDCl3): d
165, 161.7 (Cꢁ
129.7, 128.4, 128.3, 128.2, 128, 127.9, 127.8, 127.7, 96.8
(C-1), 79.3 (C-3), 74.8 (C-5), 74.7, 73.5, 72.2 (C-4), 71.7,
ꢀ
/
458 (c 1, CHCl3); H
/H), 7.65 (m,
/H),
/
/
/
/
Hz, H-2), 5.16 (t, 1 H, J 9.0 Hz, H-1), 5.0 (d, 1 H,
CH2Ph), 4.68 (m, 3 H, CH2Ph), 4.5 (d, 2 H, CH2Ph),
4.15 (d, 1 H, J3.4 2.3 Hz, H-4), 3.85 (m, 2 H, H-3, H-5),
3.7 (m, 2 H, H-6a/6b); 13C NMR (CDCl3): d 166.9,
162.1, 138, 137.6, 137.3, 133.6, 129.9, 129.1, 128.6,
128.5, 128.4, 128.3, 128, 127.9, 127.8, 80.7 (C-1), 79.6
(C-3), 75.6 (C-5), 75.1, 73.6, 72.9 (C-4), 72.3, 71.2 (C-2),
/NH), 138.1, 137.6, 137.3, 133, 129.8,
67.7 (C-6); ESIMS: m/z 722 (Mꢀ
Na)ꢀ.
/

1452
K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ1454
/
1
3.19. Allyl 2-O-p-methoxybenzoyl-3,4,6-tri-O-benzyl-b-
-galactopyranoside (30)
yellow solid; mp 88 8C; [a]D
NMR (CDCl3): d 8.33 (d, 2 H, Bzꢀ
H), 7.45ꢁ7.15 (m, 15 H), 5.78 (m, 1 H, CH ꢁ
(dd, 1 H, J2,3 10, J1,2 8.0 Hz, H-2), 5.21 (dd, 1 H, J 17.3,
J 1.3 Hz, CHꢁCH2), 5.1 (dd, 1 H, J 10.4, J 0.9 Hz,
CHꢁCH2), 5.04 (d, 1 H, CH2Ph), 4.71 (m, 2 H, CH2Ph),
4.59 (d, 1 H, H-1), 4.5 (m, 3 H, CH2Ph), 4.35 (dd, 1 H, J
13.4, J 4.6 Hz, CH2ꢀCHꢁCH2), 4.09 (m, 2 H, H-4,
CH2ꢀCHꢁ
CH2), 3.7 (m, 4 H, H-3, H-5, H-6a/b); 13C
ꢀ
/
7.78 (c 0.68, CHCl3); H
H), 8.18 (d, 2 H, Bzꢀ
CH2), 5.7
D
/
/
/
/
Allyl
2-O-acetyl-3,4,6-tri-O-benzyl-b-
D-galactopyra-
nose (26) was treated with NaOMe as indicated for
compound 27. The resulting deacetylated product (0.536
g, 1.1 mmol) in THF (15 mL) was treated with 1,3-
dicyclohexylcarbodiimide (DCC) (0.811 g, 3.9 mmol),
and the reaction was stirred for 30 min at r.t. p-
Methoxybenzoic acid (0.831 g, 5.5 mmol) was then
added followed by DMAP (38 mg, 0.3 mmol). The
reaction was refluxed for 16 h. After cooling, the solvent
was evaporated, the residue taken up in EtOAc and
filtered over Celite. The pad was rinced with EtOAc,
and the combined filtrates washed with HCl (2 N),
NaHCO3 (satd), and brine. The solution was dried
(Na2SO4), and the solvent evaporated. The crude
product was purified by column chromatography (4:1
/
/
/
/
/
/
NMR (CDCl3): d 163.4, 150.5, 138.2, 137.7, 137.5,
135.6, 133.7, 130.8, 128.5, 128.3, 128.2, 128, 127.9,
127.8, 127.7, 127.6, 123.4, 117.2, 99.8 (C-1), 79.7, (C-
3), 74.6, 73.7 (C-5), 73.6, 72.7 (C-2), 72.1 (C-4), 71.5,
69.3, 68.5 (C-6); ESIMS: m/z 662 (Mꢀ
Na)ꢀ; Anal.
Calcd for C37H37NO9: C, 69.47; H, 5.83; N, 2.19.
Found: C, 69.07; H, 5.86; N, 2.18.
/
3.21. 2-O-p-Methoxybenzoyl-3,4,6-tri-O-benzyl-a/b-D-
galactopyranosyl trichloroacetimidate (32)
cyclohexaneꢁ
white solid; mp 67 8C; [a]D
NMR (CDCl3): d 8.07 (d, 2 H, Bzꢀ
H), 7.0 (d, 2 H, BzꢀH), 5.81 (m, 1 H, CH ꢁ
(dd, 1 H, J2,3 9.5, J1,2 8.4 Hz, H-2), 5.25 (d, 1 H, J 17.3
CH2), 5.07
/
EtOAc) to give 30 (0.386 g, 57%) as a
1
ꢀ
/
18.18 (c 0.78, CHCl3); H
H), 7.45ꢁ7.2 (m, 15
CH2), 5.73
/
/
The anomeric allyl group was first removed in the same
manner as for compound 28. To a solution of the crude
alcohol (0.339 g, 0.46 mmol) and trichloroacetonitrile (2
mL) in anhyd CH2Cl2 (15 mL) was added anhyd K2CO3
(0.500 g). The mixture was stirred overnight, the solid
material was filtered off and the solvent evaporated. The
oily residue was purified by column chromatography
/
/
Hz, CHꢁ
/
CH2), 5.12 (d, 1 H, J 10.5 Hz, CHꢁ
/
(d, 1 H, CH2Ph), 4.71 (d, 2 H, CH2Ph), 4.6 (d, 1 H, H-1),
4.53 (m, 3 H, CH2Ph), 4.37 (dd, 1 H, J 13.5, J 3.5 Hz,
CH2ꢀ
/
CHꢁ
/
CH2), 4.14 (ddd, 1 H, J 13.5, J 6.2, J 0.9 Hz,
CH2), 4.08 (d, 1 H, J3,4 2.2 Hz, H-4), 3.95 (s,
CH2ꢀ
/
CHꢁ
/
(9:1 cyclohexaneꢁEtOAc, 1% Et3N) to give the a
/
3 H, OCH3), 3.72 (m, 4 H, H-3, H-5, H-6a/b); 13C NMR
(CDCl3): d 165, 163.3, 138.5, 137.9, 137.8, 133.9, 131.9,
128.5, 128.3, 128.2, 127.9, 127.8, 127.7, 127.6, 127.5,
122.7, 117, 113.5, 100.2 (C-1), 79.9 (C-3), 74.4, 73.7 (C-
5), 73.6, 72.4 (C-4), 71.7, 71.6 (C-2), 69.3, 68.7 (C-6),
trichloroacetimidate 32a as an oil (0.082 g, 19%) and
the b trichloroacetimidate 32b as an amorphous white
solid (0.265 g, 63%).
a Anomer: [a]D
(CDCl3): d 8.47 (s, 1 H, NH), 7.9 (d, 2 H, Bzꢀ
7.2 (m, 15 H), 6.95 (d, 2 H, BzꢀH), 6.69 (d, 1 H, J1,2 3.5
ꢀ
/
77.48 (c 1, CHCl3); 1H NMR
H), 7.45ꢁ
/
/
55.5; ESIMS: m/z 647 (Mꢀ
/
Na)ꢀ; Anal. Calcd for
/
C38H40O8×
/
0.2 cyclohexane: C, 73.39; H, 6.66. Found: C,
Hz, H-1), 5.85 (dd, 1 H, J2,3 10 Hz, H-2), 5.09 (d, 1 H,
CH2Ph), 4.79 (d, 1 H, CH2Ph), 4.71 (d, 2 H, CH2Ph),
4.56 (d, 1 H, CH2Ph), 4.51 (d, 1 H, CH2Ph), 4.27 (m, 3
H, H-3, H-4, H-5), 3.92 (s, 3 H, OCH3), 3.78 (dd, 1 H,
J6a,6b 9.1, J5,6a 8.0 Hz, H-6a), 3.69 (dd, 1 H, J5,6b 5.4 Hz,
73.09; H, 6.95.
3.20. Allyl 2-O-p-nitrobenzoyl-3,4,6-tri-O-benzyl-b-D-
galactopyranoside (31)
H-6b); 13C NMR (CDCl3): d 165.3, 163.5, 160.5 (Cꢁ
/
Allyl
2-O-acetyl-3,4,6-tri-O-benzyl-b-
D-galactopyra-
NH), 138.3, 137.7, 131.9, 128.5, 128.4, 128.3, 128.2, 128,
127.9, 127.8, 127.7, 122, 113.5, 94.7 (C-1), 75.6 (C-3),
74.9, 73.6 (C-4), 73.5, 72.3 (C-5), 72.2, 69.8 (C-2), 68.1
(C-6), 55.4.
nose (26) was treated with NaOMe as indicated for
compound 27. The resulting de-acetylated product
(0.498 g, 1.0 mmol) in THF (13 mL) was treated with
DCC (0.754 g, 3.7 mmol), and the reaction was stirred
for 30 min at r.t. p-Nitrobenzoic acid (0.848 g, 5.1
mmol) was then added followed by DMAP (34 mg, 0.3
mmol). Stirring was continued for 30 min when TLC
indicated complete conversion of the starting material.
The solvent was evaporated, the residue taken up in
EtOAc and filtered over Celite. The pad was washed
with EtOAc, and the combined filtrates washed with
HCl (2 N), NaHCO3 (satd), and brine. The solution was
dried (Na2SO4), and the solvent evaporated. The crude
product was purified by column chromatography (4:1
b Anomer: [a]D
(CDCl3): d 8.58 (s, 1 H, NH), 7.98 (d, 2 H, Bzꢀ
7.45ꢁ7.2 (m, 15 H), 6.95 (d, 2 H, BzꢀH), 5.95 (t, 1 H,
J1,2 J2,3 8.2 Hz, H-2), 5.89 (d, 1 H, H-1), 5.09 (d, 1 H,
CH2Ph), 4.71 (d, 2 H, CH2Ph), 4.6ꢁ4.5 (m, 3 H,
CH2Ph), 4.14 (d, 1 H, J3,4 1.8 Hz, H-4), 3.93 (s, 3 H,
OCH3), 3.9 (t, 1 H, J5,6a J5,6b 6.7 Hz, H-5), 3.81ꢁ3.71
(m, 3 H, H-3, H-6a/b); 13C NMR (CDCl3): d 164.7,
NH), 138.2, 137.7, 137.5, 131.8, 128.5,
ꢀ
/
418 (c 1, CHCl3); 1H NMR
H),
/
/
/
ꢃ
/
/
ꢃ
/
/
163.4, 161.8 (Cꢁ
/
128.4, 128.3, 128.1, 127.9, 127.8, 122.3, 113.5, 96.9 (C-1),
79.3 (C-3), 74.8 (C-5), 74.7, 73.6, 72.3 (C-4), 71.7, 70.3
cyclohexaneꢁ
/
EtOAc) to give 31 (0.587 g, 90%) as a pale
(C-2), 68 (C-6), 55.4; Anal. Calcd for C37H36Cl3N1O7×
/

K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ
/1454
1453
0.8 cyclohexane: C, 63.04; H, 5.77; N, 1.76. Found: C,
62.97; H, 5.72; N, 1.81.
3.23.2. Using the b-glycosyl trichloroacetimidate. Cou-
pling of the acceptor 9 (25 mg, 74 mmol) and donor 32b
(81 mg, 111 mmol) using TMSOTf (37 mL, 3.7 mmol)
3.22. 2-O-p-Nitrobenzoyl-3,4,6-tri-O-benzyl-a/b-
galactopyranosyl trichloroacetimidate (33)
D-
according to the general procedure at ꢂ
gave, after purification (1.5:1 cyclohexaneꢁ
disaccharide 34 (0.033 g, 47%) as well as the rearrange-
ment product (0.026 g, 32%). Reaction at ꢂ78 8C (2 h)
/
20 8C for 20 min
/EtOAc) the
The anomeric allyl group was first removed in the same
manner as for compound 28. To a solution of the crude
alcohol (0.356 g, 0.59 mmol) and trichloroacetonitrile (2
mL) in anhyd CH2Cl2 (15 mL) was added anhyd K2CO3
(0.500 g). The mixture was stirred overnight, the solid
material was filtered off and the solvent evaporated. The
oily residue was purified by column chromatography
/
gave 62% of the disaccharide and 30% of the rearrange-
ment product.
Disaccharide 34: mp 127 8C; [a]D
ꢀ
/
19.28 (c 1,
CHCl3); H NMR (CDCl3): d 8.02 (d, 2 H, BzꢀH),
7.42ꢁ7.18 (m, 15 H), 6.98 (d, 2 H, BzꢀH), 5.87 (m, 1 H,
CH ꢁCH2), 5.65 (dd, 1 H, J2?,3? 9.8, J1?,2? 8.2 Hz, H-2?),
5.25 (dd, 1 H, J 17.2, J 1.3 Hz, CHꢁCH2), 5.1 (d, 1 H, J
10.5 Hz, CHꢁCH2), 5.06 (d, 1 H, CH2Ph), 4.9 (d, 1 H,
H-1?), 4.69 (m, 4 H, ꢀOCH2Oꢀ, CH2Ph), 4.58 (m, 2 H,
H-1, ꢀOCH2Oꢀ), 4.5 (d, 1 H, CH2Ph), 4.46 (m, 3 H, ꢀ
OCH2Oꢀ, CH2Ph), 4.35 (dd, 1 H, J 12.4, J 5.3 Hz,
CH2ꢀCHꢁCH2), 4.05 (m, 2 H, H-4?, CH2ꢀCHꢁCH2),
1
/
/
/
/
(6:1 cyclohexaneꢁ
/
EtOAc, 1% Et3N) to give the a
/
trichloroacetimidate 33a as an oil (0.172 g, 39%) and
the b trichloroacetimidate 33b as an amorphous white
solid (0.115 g, 26%).
/
/
/
/
/
/
a Anomer: [a]D
(CDCl3): d 8.45 (s, 1 H, NH), 8.3 (d, 2 H, Bzꢀ
2 H, BzꢀH), 7.45ꢁ7.26 (m, 15 H), 6.7 (d, 1 H, J1,2 3.5
ꢀ
/
97.68 (c 1, CHCl3); 1H NMR
H), 8.1 (d,
/
/
/
/
/
/
/
/
3.95 (s, 3 H, OCH3), 3.89 (d, 1 H, J4,5 9.2 Hz, H-5), 3.8
(s, 3 H, OCH3), 3.71 (m, 2 H, H-4, H-6a?), 3.61 (m, 5 H,
H-2, H-3, H-3?, H-5?, H-6b?), 3.27 (s, 3 H, OCH3), 3.23
(s, 3 H, OCH3); 13C NMR (CDCl3): d 168.9, 165, 163.3,
138.4, 137.6, 137.5, 133.7, 131.8, 128.4, 128.3, 128.2,
128, 127.9, 127.6, 127.5, 122.7, 117.1, 113.6, 101.9 (C-1),
101.3 (C-1?), 98.6, 98.2, 81.8 (C-2), 80.1 (C-3?), 79.8 (C-
3), 77.1 (C-4), 74.9 (C-5), 74.4, 73.6, 73.5 (C-5?), 72.3 (C-
2?), 72.1 (C-4?), 71.5, 70.6, 68.3 (C-6?), 56.2, 56.1, 55.4,
Hz, H-1), 5.86 (dd, 1 H, J2,3 10.3 Hz, H-2), 5.08 (d, 1 H,
CH2Ph), 4.79 (d, 1 H, CH2Ph), 4.71 (d, 1 H, CH2Ph),
4.61 (d, 1 H, CH2Ph), 4.56 (d, 1 H, CH2Ph), 4.51 (d, 1 H,
CH2Ph), 4.29 (m, 3 H, H-3, H-4, H-5), 3.79 (dd, 1 H,
J6a,6b 9.0, J5,6a 8.0 Hz, H-6a), 3.7 (dd, 1 H, J5,6b 5.4 Hz,
H-6b); 13C NMR (CDCl3): d 163.7, 160.6 (Cꢁ
/NH),
150.6, 138.1, 137.6, 137.4, 134.9, 130.8, 128.5, 128.4,
128.3, 128.2, 128.1, 128, 127.9, 127.8, 123.5, 94.3 (C-1),
75.2 (C-3), 75.1, 73.6, 73.2 (C-4), 72.4 (C-5), 71.8, 70.7
52.4; ESIMS: m/z 925 (Mꢀ
Na)ꢀ; Anal. Calcd for
C49H58O16: C, 65.18; H, 6.47. Found: C, 64.79; H, 6.43.
/
(C-2), 68 (C-6).
b Anomer: [a]D
1
ꢀ
/
48.88 (c 0.67, CHCl3); H NMR
(CDCl3): d 8.62 (s, 1 H, NH), 8.3 (d, 2 H, Bzꢀ
/
H), 8.11
3.24. Methyl (2-O-p-nitrobenzoyl-3,4,6-tri-O-benzyl-b-
(d, 2 H, BzꢀH), 7.46ꢁ7.15 (m, 15 H), 5.93 (m, 2 H, H-1,
/
/
D
-galactopyranosyl)-(10
/
2)-(allyl 3,4-di-O-
H-2), 5.06 (d, 1 H, CH2Ph), 4.73 (m, 2 H, CH2Ph), 4.54
(m, 3 H, CH2Ph), 4.2 (d, 1 H, J3,4 2.0 Hz, H-4), 3.92 (m,
methoxymethyl-b-D-glucopyranosid)uronate (35)
1 H, H-5), 3.83ꢁ
/
3.72 (m, 3 H, H-3, H-6a/b); 13C NMR
3.24.1. Using the a-glycosyl trichloroacetimidate. Cou-
pling of the acceptor 9 (27 mg, 80 mmol) and donor 33a
(90 mg, 120 mmol) using TMSOTf (40 mL, 4.0 mmol)
(CDCl3): d 163.2, 161.5 (Cꢁ/NH), 150.5, 138, 137.6,
137.2, 135.1, 130.8, 128.5, 128.4, 128.3, 128.1, 128,
127.9, 127.8, 123.5, 96.5 (C-1), 79.1 (C-3), 74.9, 74.8
(C-5), 73.6, 72 (C-4), 71.6, 71.5 (C-2), 67.8 (C-6); Anal.
according to the general procedure at ꢂ
gave, after purification (32:1 to 6:1 tolueneꢁ
disaccharide 35 (0.016 g, 22%) as well as the rearrange-
ment product (0.059 g, 66%). Reaction at ꢂ78 8C (20
/
20 8C for 30 min
/EtOAc) the
Calcd for C36H33Cl3N2O7×
/
0.8 cyclohexane: C, 60.40; H,
5.29; N, 3.45. Found: C, 60.35; H, 5.43; N, 3.47.
/
min) gave 24% of the disaccharide and 78% of the
rearrangement product.
3.23. Methyl (2-O-p-methoxybenzoyl-3,4,6-tri-O-
benzyl-b-
D
-galactopyranosyl)-(10
/
2)-(allyl 3,4-di-O-
methoxymethyl-b-D-glucopyranosid)uronate (34)
3.24.2. Using the b-glycosyl trichloroacetimidate. Cou-
pling of the acceptor 9 (26 mg, 77 mmol) and donor 33b
(86 mg, 116 mmol) using TMSOTf (38 mL, 3.8 mmol)
3.23.1. Using the a-glycosyl trichloroacetimidate. Cou-
pling of the acceptor 9 (25 mg, 74 mmol) and donor 32a
(81 mg, 111 mmol) using TMSOTf (37 mL, 3.7 mmol)
according to the general procedure at ꢂ
gave, after purification (32:1 to 6:1 tolueneꢁ
disaccharide 35 (0.037 g, 51%) as well as the rearrange-
ment product (0.024 g, 28%). Reaction at ꢂ78 8C (20
/
20 8C for 20 min
/EtOAc) the
according to the general procedure at ꢂ
/
20 8C for 30 min
EtOAc), the
gave, after purification (1.5:1 cyclohexaneꢁ
/
/
disaccharide 34 (0.010 g, 24%) as a white powder as well
as the trichloroacetamide rearrangement product (0.065
min) gave 80% of the disaccharide and 20% of the
rearrangement product.
g, 80%). Reaction at ꢂ
disaccharide and 70% of the rearrangement product.
/
78 8C (2 h) gave 21% of the
Disaccharide 35: mp 134 8C; [a]D
CHCl3); H NMR (CDCl3): d 8.33 (d, 2 H, BzꢀH),
ꢀ
/
27.48 (c 0.8,
1
/

1454
K. Ple´ / Carbohydrate Research 338 (2003) 1441ꢁ1454
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7. (a) Yoshiki, Y.; Okubo, K. Biosci. Biotech. Biochem. 1995,
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(b) Yoshiki, Y.; Kahara, T.; Okubo, K.; Sakabe, T.;
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8. Berhow, M.; Wagner, E.; Vaughn, S.; Plewa, M. Mutat.
8.17 (d, 2 H, Bzꢀ
CH ꢁCH2), 5.65 (dd, 1 H, J2?,3? 10, J1?,2? 8.1 Hz, H-2?),
5.25 (dd, 1 H, J 17.3, J 1.6 Hz, CHꢁCH2), 5.14 (dd, 1
H, J 10.5, J 1.4 Hz, CHꢁCH2), 5.05 (d, 1 H, CH2Ph),
4.92 (d, 1 H, H-1?), 4.71ꢁ4.62 (m, 4 H, ꢀOCH2Oꢀ
CH2Ph), 4.55 (m, 2 H, H-1, ꢀOCH2Oꢀ), 4.5ꢁ4.4 (m, 4
H, ꢀOCH2Oꢀ, CH2Ph), 4.35 (ddt, 1 H, J 12.4, J 5.1, J
1.5 Hz, CH2ꢀCHꢁCH2), 4.09 (d, 1 H, J3?,4? 2.2 Hz, H-
4?), 4.05 (dd, 1 H, J 12.4, J 6.0 Hz, CH2ꢀCHꢁCH2),
3.88 (d, 1 H, J4,5 9.1 Hz, H-5), 3.8 (s, 3 H, OCH3), 3.74ꢁ
/
H), 7.45ꢁ
/
7.15 (m, 15 H), 5.88 (m, 1 H,
/
/
/
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/
/,
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Res. 2000, 448, 11ꢁ
9. (a) Garegg, P. J.; Olsson, L.; Oscarson, S. J. Org. Chem.
1995, 60, 2200ꢁ2204;
/
22.
/
/
/
/
/
/
/
(b) Battistelli, C. L.; De Castro, C.; Iadonisi, A.; Lanzetta,
R.; Mangoni, L.; Parrilli, M. J. Carbohydr. Chem. 1999,
/
3.6 (m, 6 H, H-3?, H-3, H-4, H-5?, H-6a/b?), 3.58 (dd, 1
H, J 7.1, J 6.9 Hz, H-2), 3.25 (s, 3 H, OCH3), 3.23 (s, 3
H, OCH3); 13C NMR (CDCl3): d 168.8, 163.4, 150.5,
138.2, 137.5, 137.3, 135.6, 133.6, 130.7, 128.5, 128.3,
128.2, 128, 127.9, 127.8, 127.6, 123.5, 117.2, 101.7 (C-1),
101 (C-1?), 98.3, 98.2, 81.6 (C-2), 79.8 (C-3?), 79.3 (C-3),
77.2 (C-4), 75 (C-5), 74.6, 73.6 (C-2?, C-5?), 71.8 (C-4?),
71.3, 70.6, 68.1 (C-6?), 56.2, 56.1, 52.4; ESIMS: m/z 940
18, 69ꢁ
10. Saito, S.; Ichinose, K.; Sasaki, Y.; Sumita, S. Chem.
Pharm. Bull. 1992, 40, 3261ꢁ3268.
11. (a) Saito, S.; Kuroda, K.; Hayashi, Y.; Sasaki, Y.;
Nagamura, Y.; Nishida, K.; Ishiguro, I. Chem. Pharm.
/
86.
/
Bull. 1991, 39, 2333ꢁ2339;
/
(b) Saito, S.; Sasaki, Y.; Kuroda, K.; Hayashi, Y.; Sumita,
S.; Nagamura, Y.; Nishida, K.; Ishiguro, I. Chem. Pharm.
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/543;
(Mꢀ
Na)ꢀ; Anal. Calcd for C48H55NO17: C, 62.81; H,
6.04; N, 1.53. Found: C, 62.86; H, 6.16; N, 1.61.
/
/
/
Acknowledgements
/
1804.
The author thanks the C.N.R.S. (Centre National de
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/
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