Synthesis of ABO histo-blood group type i and II antigens

Article abstract of DOI:10.1016/j.carres.2010.08.012

The ABO histo-blood group system is one of the most clinically important antigen families. As part of our overall goal to prepare the entire set of the A, B and H type I-VI antigens for a range of biochemical investigations, we report herein the synthesis of the type I and II antigens with a 7-octen-1-yl aglycone. This linker was chosen to facilitate not only the future conjugation of the antigens to a protein or solid support but also the synthesis of the H type I and II octyl glycosides for enzyme kinetic studies.

Full text of DOI:10.1016/j.carres.2010.08.012

Carbohydrate Research 345 (2010) 2305–2322
Contents lists available at ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Synthesis of ABO histo-blood group type I and II antigens
⇑
Peter J. Meloncelli, Todd L. Lowary
Department of Chemistry and Alberta Ingenuity Centre for Carbohydrate Science, Gunning-Lemieux Chemistry Centre, University of Alberta, Edmonton, AB, Canada T6G 2G2
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 25 June 2010
Received in revised form 13 August 2010
Accepted 17 August 2010
Available online 16 September 2010
The ABO histo-blood group system is one of the most clinically important antigen families. As part of our
overall goal to prepare the entire set of the A, B and H type I–VI antigens for a range of biochemical inves-
tigations, we report herein the synthesis of the type I and II antigens with a 7-octen-1-yl aglycone. This
linker was chosen to facilitate not only the future conjugation of the antigens to a protein or solid support
but also the synthesis of the H type I and II octyl glycosides for enzyme kinetic studies.
Ó 2010 Elsevier Ltd. All rights reserved.
Keywords:
ABO histo-blood group
ABO type I
ABO type II
Chemical synthesis
Trichloroacetimidate
7-Octen-1-yl
1. Introduction
the nomenclature of the type V and VI antigens was not clear.10
Although a bit tangential to the main goal of this paper, we de-
The ABO histo-blood group system has long been of interest to
the biological and medical sciences as these antigens are one of the
most important clinical considerations for both transfusion and
transplantation.1,2 This family of antigens consists of three oligo-
saccharide epitopes (1–3, Fig. 1), the structures of which were elu-
cidated by Morgan and Watkins in 1957.3 The H(O) antigen was
shown to be a disaccharide consisting of L-fucose and D-galactose
(3, Fig. 1), while the A and the B antigens (1 and 2) were shown
to be trisaccharides containing the disaccharide H antigen with
scribe briefly this ambiguity, as a concise description does not ap-
pear to be present elsewhere in the literature. The first report of a
type VI antigen structure from human sources was achieved in
1956 by Kuhn and co-workers, who isolated the H type VI antigen
as the free oligosaccharide in appreciable quantities from human
milk.11 Subsequently, a type VI antigen from tissue samples was
reported by Briemer and co-workers in 1982.12 This glycolipid,
named A-4 by Briemer, contained what was in the future termed
the A type VI antigen, linked to ceramide.
an additional N-acetyl-galactosamine or
a
galactose residue,
The origin of the type V antigen proved more difficult to un-
cover. The first mention of this structure was by Mollicone and
co-workers in 1990.8 The authors obtained synthetic samples of
the type V and VI antigens from Chembiomed, a now defunct com-
mercial entity, and this is presumably the origin of this nomencla-
ture. To the best of our knowledge, no ABH type V structure has
been isolated from a mammalian source. A recent review by
Yamamoto on the ABO histo-blood group system makes reference
to the type I–IV and type VI antigens, excluding the type V antigen.1
This exclusion of the type V antigen was likely based on the lack of
evidence for existence of ABH type V structures in humans. This is
surprising because Hakomori and Stellner reported the isolation of
a glycolipid from human erythrocytes that contained the type V
respectively. It was later shown by Tuppy and Staudenbauer that
individuals who are of the A histo-blood group possess an N-acet-
yl-galactosaminyl transferase (GTA) capable of adding N-acetyl
galactosamine to the 3-OH group of the H antigen galactose moi-
ety.4 On the other hand, individuals of the B histo-blood group pos-
sess a galactosyl transferase (GTB) that adds galactose to this
hydroxyl group. The genetic basis of the ABO histo-blood group
system was discovered in the 1990s, when the genes responsible
for coding GTA and GTB were identified.5–7
The ABO antigens can be further divided into six subtypes, type
I–VI (Table 1).8 The type I–IV and VI antigens are present either as
glycoproteins, glycolipids, or as free oligosaccharides in humans.1
After the publication of our synthesis of the ABO type V and VI his-
to-blood group antigens,9 it came to our attention that the origin of
(b-D-Galp-(1?3)-b-D
-Gal) backbone at the terminal end.13 This
structure could easily be converted into the H type V antigen using
the same fucosyl transferase responsible for the biosynthesis of the
H type I–IV antigens present on erythrocytes. Based on this
chronology, the nomenclature assignment of the type V and VI anti-
gens should be reversed; however, as the nomenclature is well
⇑
Corresponding author. Tel.: +1 780 492 1861; fax: +1 780 492 7705.
E-mail address: tlowary@ualberta.ca (T.L. Lowary).
0008-6215/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2010.08.012

2306
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
HO
OH
HO
HO
OH
O
OH
O
HO
HO
O
OH
O
OH
HO
O
HO
O
OR
HO
HO
AcHN
O
OR
O
OR
O
O
O
OH
O
O
OH
OH
OH
HO
OH
OH
HO
HO
H Antigen
B Antigen
A Antigen
3
1
2
Figure 1. Structures of the A, B and H(O) antigens. R = Glycoprotein, glycolipid or free oligosaccharide.
Table 1
2. Results and discussion
Carbohydrate moieties responsible for the six subtypes (I–VI) of the ABO histo-blood
group antigens
To prepare 4–9 we chose a linear chemical synthesis, similar to
the strategy employed in our recently reported synthesis of the
type V and VI antigens.9 This strategy would enable an expedited
synthesis of all three of the type I and II antigens in sufficient quan-
tities for a range of investigations. A block synthesis, such as that
reported by Bovin and co-workers,17 would not enable access to
the H structures. We chose to employ a 7-octen-1-yl aglycone for
two reasons. Firstly, it would enable ready access to the octyl gly-
coside, via catalytic hydrogenation, for enzyme kinetics studies.24
Secondly, it would enable facile preparation of glycoconjugates.
We have demonstrated that a thiol-ene ‘click reaction’25 can be
used to introduce a reactive linker that allows direct conjugation
of carbohydrates to silica-based surfaces.26
Type
Carbohydrate
Type I
ABO-b-(1?3)-b-
ABO-b-(1?4)-b-
ABO-b-(1?3)-a-D-GalpNAc
ABO-b-(1?3)-b-
ABO-b-(1?3)-b-
ABO-b-(1?4)-b-
D
-GlcpNAc
-GlcpNAc
Type II
Type III
Type IV
Type V
Type VI
D
D
D
D
-GalpNAc
-Galp
-Glcp
established in multiple sources we have adhered to the conven-
tional naming of the type I–VI antigens.
The overall goal of our synthetic endeavour is to prepare all
known ABO type I–VI histo-blood group antigens. We hope that
with all of the antigens in hand, we will be able to conduct a range
of studies on the biological importance of these sub-types. An
example of such an investigation is quantitatively determining
the difference between the A1 and A2 blood groups. It has been
shown that individuals of the A1 subtype have a higher density of
A antigen on their erythrocytes than individuals of the A2 subtype.14
Further work by Clausen and co-workers has demonstrated that this
difference can be attributed to the fact that GTA1 is able to transfer
N-acetyl-galactosamine onto the H type I–IV structures present on
erythrocytes, while GTA2 is only able to affect transfer onto the H
type I and type II structures.15 These pioneering studies were con-
ducted in a qualitative manner via immunostaining. Access to syn-
thetic samples of these antigens would enable more detailed
quantitative kinetics assays to be conducted.
2.1. Synthesis of type I antigens
The synthesis of the type I ABO histo-blood group antigens com-
menced from the known27 and easily prepared trichloroacetimi-
date donor 10 (Scheme 1). Glycosylation with the 7-octen-1-ol
acceptor afforded the 7-octen-1-yl glycoside 11 as a mixture of
anomers (a/b, 11:41). In the absence of a C2 participating group,
the stereochemical outcome for this glycosylation was dependent
on the anomeric configuration of the trichloroacetimidate. To en-
sure a high ratio of the desired b-anomer, a trichloroacetimidate
donor with predominately the
a stereochemistry was required.
Presumably, the mechanism for the formation of the b-glycoside
involves an SN2-type process.28 Introduction of the aglycone in
the presence of an azido protecting group was not trivial when
compared to other amine protecting groups such as N-phthali-
mide; however, the azido group can be converted into an amine
under milder conditions than the phthalimide. Deacetylation with
sodium methoxide, followed by introduction of a benzylidene ace-
Herein we report the synthesis of the ABO type I and II histo-
blood group antigens (4–9, Fig. 2) attached to an octen-1-yl agly-
cone. To date, the synthesis of the ABO type I and II antigens has
received more interest than the type III–VI structures. The chemi-
cal synthesis of all three type I antigens has been reported by Paul-
tal, enabled isolation of the corresponding b and
and 14, respectively, in moderate yield, with the desired b anomer
being the predominant product. The 1H NMR spectrum of the
a glycosides, 13
sen and Kolár as well as by Bovin and co-workers.17,18 The
ˇ16
chemoenzymatic synthesis of the A type I and II tetrasaccharides
was reported by Palcic and co-workers,19 and more recently Wang
and co-workers have developed a chemoenzymatic synthesis of
the B type I antigen.20 Interestingly, the latter study did not use
the human glycosyltransferases, but rather three enzymes present
in Escherichia coli O86, which produces a B type III mimetic in its
lipopolysaccharide O-chain. The synthesis of the type II antigens
has been more limited, most notably the chemical synthesis of
the ABO type II structures was achieved by Paulsen and co-work-
ers21,22 as well as by Bovin and co-workers.23
a
anomer 13 showed an anomeric proton signal appearing at
4.90 ppm as a doublet with a coupling of 3.6 Hz, while the spec-
trum of the b anomer 14 had an anomeric proton signal at
4.42 ppm as a doublet with a coupling of 8.0 Hz.
Introduction of the galactose core residue was achieved by
glycosylation of 13 with trichloroacetimidate 15 affording disac-
charide 16 (Scheme 2). To facilitate removal of trichloroacetimi-
date-related impurities, a deacetylation was conducted under
Zemplén conditions to afford diol 17 in moderate yield (63%) over
two steps. Introduction of the pivaloyl ester selectively at the 30
position of 17 was achieved with complete regioselectivity using
trimethylacetyl chloride in pyridine in excellent yield (89%). Next,
The A1 and A2 blood groups should not be confused with the A type I and type II
structures. The latter refers to distinct chemical entities while the former refers to
subclasses of the A blood type, which are characterized by different densities of the
antigen on erythrocytes.
introduction of the
a-
L-fucopyranosyl moiety was achieved
through the use of b-
L-fucopyranosyl trichloroacetimidate 19

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2307
HO
OH
HO
OH
O
O
OH
OH
OH
OH
OH
O
OH
O
O
HO
HO
O
HO
HO
AcHN
HO
O
O
O
O
O
O
NHAc
NHAc
O
O
O
O
OH
OH
4
5
OH
OH
B Type I
HO
A Type I
HO
HO
OH
O
HO
OH
OH
O
OH
O
HO
O
O
O
HO
O
HO
HO
O
NHAc
O
O
NHAc
O
O
OH
OH
7
6
OH
OH
H Type I
H Type II
HO
HO
HO
HO
HO
OH
O
OH
O
OH
OH
OH
OH
HO
OH
O
OH
O
O
O
AcHN
HO
O
O
HO
O
O
HO
O
O
O
O
NHAc
NHAc
O
O
OH
OH
OH
8
9
OH
HO
HO
B Type II
A Type II
Figure 2. Structure of ABO histo-blood group type I and II antigen targets.
OAc
O
OAc
O
NH
(a)
AcO
AcO
AcO
AcO
CCl3
O
N3
O
N3
10
11
(b)
O
Ph
O
O
O
HO
N3
OH
O
(c)
13
14
HO
HO
O
N3
12
O
Ph
O
O
HO
N3
O
Scheme 1. Reagents: (a) HO(CH2)6CH@CH2, 4 Å MS, TMSOTf, CH2Cl2; (b) NaOCH3, CH3OH; (c) PhCH(OCH3)2, p-TsOH, DMF, (a/b, 11:41) 52% (three steps).
following the methodology of Schmidt and co-workers.29 Treat-
ment of 18 with trichloroacetimidate 19 under standard glycosyl-
ation conditions afforded trisaccharide 20 in good yield (80%) and
with excellent stereoselectivity. The anomeric proton of the fuco-
pyranoside of 20 showed an anomeric signal at 5.41 ppm as a dou-
blet with a coupling constant of 1.5 Hz. Removal of the pivaloate
ester from 20 required forcing conditions (lithium methoxide in
methanol at reflux) and prolonged reaction times (seven days);
however, the reaction afforded alcohol 21 in good yield. A similar
observation was made in our synthesis of the type V and VI anti-
gens. The decision to continue using this protecting group as op-
posed to others that might be removed more efficiently was
based on the fact that the pivaloate ester could be installed and re-
moved in good yield and with minimal by-products.9
Access to the H type I antigen was achieved first by conversion of
the azide moiety into an N-acetyl group using thiolacetic acid in pyr-
idine to afford 22 (Scheme 3). Surprisingly, some acetylation of the
alcohol in 22 was observed during this reaction. The formation of
this by-product was not, however, problematic as Zemplén deacet-
ylation easily converted the O-acetylated compound into 22.
Next, global deprotection was achieved using a Birch reduction
resulting in the conversion of 22 into 6 in 92% yield. For investiga-

2308
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
O
Ph
O
O
O
HO
N3
Ph
Ph
13
O
O
Ph
O
O
O
(a)
O
Ph
O
Ph
O
O
O
O
O
O
O
O
O
O
O
RO
PivO
N3
N3
OR
(b)
HO
O
18
16 R = Ac
17 R = H
AcO
(c)
AcO
O
CCl3
NH
15
O
OBn
CCl3
O
NH
(d)
OBn
BnO
19
Ph
Ph
O
O
O
O
O
Ph
O
O
Ph
O
O
O
O
O
O
O
O
O
(e)
HO
PivO
N3
N3
O
O
O
O
OBn
OBn
OBn
OBn
21
BnO
20
BnO
Scheme 2. Reagents: (a) TMSOTf, 4 Å MS, CH2Cl2; (b) NaOCH3, CH3OH, 63% (two steps); (c) (CH3)3CCOCl, pyridine, 89%; (d) TMSOTf, 4 Å MS, Et2O, CH2Cl2, 80%; (e) LiOCH3,
CH3OH, 82%.
Ph
Ph
O
O
O
O
O
Ph
O
Ph
O
O
O
O
O
O
O
O
O
HO
(a)
O
HO
N3
O
NHAc
O
O
O
OBn
OBn
22
21
OBn
OBn
BnO
BnO
(b)
OH
O
HO
OH
O
HO
OH
OH
O
HO
O
HO
O(CH2)7CH3
(c)
O
O
O
HO
HO
NHAc
NHAc
O
O
O
O
OH
23
OH
OH
OH
HO
HO
6
Scheme 3. Reagents: (a) (i) CH3COSH, pyridine; (ii) NaOCH3, CH3OH, 62%; (b) NH3, Na, CH3OH, THF, 92%; (c) H2, 10% Pd–C, THF, H2O, 83%.
tions involving conjugation of the H type I antigen to a support, the
7-octen-1-yl glycoside 6 was desired; however, other studies, such
as the examination of GTA and GTB kinetics, required the octyl gly-
coside. To this end, catalytic reduction of the alkene moiety of 6
afforded octyl glycoside 23 in 83% yield.
Access to the A type I antigen was achieved first by glycosyla-
tion of 21 with trichloroacetimidate 24 to afford tetrasaccharide
25 in good (92%) yield (Scheme 4). The stereochemistry of the
newly formed glycosidic linkage was confirmed using 1H NMR
spectroscopy; the anomeric proton of the 2-azido-2-deoxy-a-D-
galactopyranoside appeared at 5.88 ppm as a doublet with a cou-
pling of 3.2 Hz. Conversion of the two azides into N-acetyl groups
using thiolacetic acid in pyridine afforded 26. Deacetylation under
Zemplén conditions afforded 27 and subsequent complete depro-
tection using dissolving metal reduction afforded 5. Overall, tetra-
saccharide 5 was obtained in 69% yield from 25.
The B type I antigen could be obtained first by glycosylation of
21 with trichloroacetimidate 28 affording tetrasaccharide 29 in

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2309
Ph
O
O
O
Ph
O
O
Ph
O
O
O
HO
O
AcO
N3
OAc
O
O
O
O
(a)
Ph
O
O
AcO
OBn
O
O
N3
O
OBn
O
21
24
O
BnO
N3
O
AcO
AcO
O
OAc
O
OBn
25
OBn
O
CCl3
BnO
N3
NH
(b)
Ph
RO
O
HO
HO
OR
O
OH
O
O
OH
OH
OH
O
O
Ph
RO
O
O
O
HO
(d)
O
AcHN
AcHN
O
O
O
O
O
O
NHAc
NHAc
O
O
O
O
OBn
OH
OBn
OH
BnO
HO
26 R = Ac
27 R = H
5
(c)
Scheme 4. Reagents: (a) TMSOTf, 4 Å MS, Et2O, 92%; (b) CH3COSH, pyridine, 74%; (c) NaOCH3, CH3OH, 94%; (d) NH3, Na, CH3OH, THF, 88%.
Ph
O
O
O
Ph
O
O
Ph
O
O
O
HO
O
BnO
BnO
N3
OBn
O
O
O
O
(a)
Ph
O
O
OBn
21
O
O
BnO
O
OBn
O
O
BnO
N3
O
BnO
BnO
O
OBn
O
OBn
29
OBn
O
CCl3
BnO
OBn
NH
28
(b)
Ph
HO
BnO
BnO
O
OH
O
OBn
O
O
OH
OH
OH
O
O
Ph
HO
O
O
HO
O
(c)
O
HO
O
BnO
O
O
O
O
O
NHAc
NHAc
O
O
O
O
OH
OBn
OH
OBn
30
4
HO
BnO
Scheme 5. Reagents: (a) TMSOTf, 4 Å MS, Et2O, 73%; (b) CH3COSH, pyridine, 65%; (c) NH3, Na, CH3OH, THF, 94%.
moderate yield (Scheme 5). Conversion of the azide into an N-acet-
yl group using thiolacetic acid in pyridine afforded 30, under con-
ditions analogous to those used in the preparation of 22. Other
techniques do exist to convert an azide into an N-acetyl group,
and several were explored including Staudinger reaction
followed by acetylation;30 however, none were found to be as
a

2310
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
efficient as the tandem reduction–acetylation process induced by
treatment with thiolacetic acid. Final deprotection of 30 using a
Birch reduction afforded the B type I antigen 4 in 94% yield. In
our synthesis of the type V and VI antigens,9 the yield for deprotec-
tion reactions of this type on similar substrates was at times low
(60–70%). These lower yields were caused by precipitation of the
starting material in liquid ammonia, leading to incomplete re-
moval of the protecting groups. In the course of the synthesis of
the targets reported here, we found that increasing the volume of
the THF used to dissolve the starting material successfully circum-
vented this problem.
type II antigen 7 with the 7-octen-1-yl glycoside moiety intact.
Reduction of part of this material by hydrogenation afforded, in
96% yield, the octyl glycoside 40 for use in enzymatic studies.
The B type II antigen was accessed first by treatment of 38 with
the trichloroacetimidate donor 28 under standard conditions
(Scheme 9). Unfortunately, complete purification of 41 was not
possible; therefore, the unequivocal confirmation of the stereo-
chemistry was not possible until after the conversion of 41 into
42 using thiolacetic acid in pyridine. In 1H NMR spectrum of 42,
the anomeric signal for the
a-galactopyranosyl linkage appeared
at 5.36 ppm as a doublet with a coupling constant of 3.6 Hz, as ex-
pected for this stereochemistry. Final global deprotection of 42
upon treatment with sodium and liquid ammonia in THF afforded
8 in high yield (96%).
2.2. Synthesis of type II antigens
The type I and II antigens are quite similar in structure. The only
difference is that the galactose core residue is linked to the reduc-
ing-end GlcNAc moiety by either a b-(1?3) or b-(1?4) linkage in
the type I and II antigens, respectively. This led us to exploit the
reducing end acceptor 13, which was used in the type I synthesis,
and putting it to use in the preparation of the type II antigens
(Scheme 6). Protection of 13 using standard benzylation conditions
afforded 31 in excellent yield, and then selective ring opening of
the benzylidene acetal using trifluoroacetic acid and triethylsilane
resulted in formation of the 4-OH derivative 32. Confirmation of
the regiochemistry of the ring opening was achieved by acetylation
(acetic anhydride and DMAP in pyridine) and observance of a
downfield shift of H-4 in the 1H NMR spectrum from 3.63 ppm in
32 to 5.05–4.91 ppm in 33.
Finally, access to the A type II antigen was achieved in an anal-
ogous manner to its A type I counterpart (Scheme 10). Glycosyla-
tion of 38 with trichloroacetimidate 24 afforded tetrasaccharide
43; subsequent reduction–acetylation of the two azides using thi-
olacetic acid in pyridine afforded 44. Confirmation of the stereo-
chemistry of the glycosidic linkage was achieved using 1H NMR
spectroscopy. The anomeric proton of the newly formed glycosidic
linkage appeared as a doublet at 5.09 ppm with a 3.7 Hz coupling
constant. Deacetylation of 44 to provide 45 followed by complete
deprotection resulted in the preparation of the A type II antigen
9 in excellent yield (88% yield over the two steps).
2.3. NMR analysis of 4–9
Next, treatment of glycosyl acceptor 32 with trichloroacetimi-
date 15 using a TMSOTf promoter system yielded an impure disac-
charide 34. Contamination of 34 with trichloroacetimidate-derived
by-products necessitated the conversion of diacetate 34 into diol
35 to facilitate purification (Scheme 7). Protection of the 30 position
of the galactose core was again achieved using trimethylacetyl
chloride in pyridine to yield 36. Glycosylation using the b-L-fuco-
pyranosyl trichloroacetimidate 19 afforded the trisaccharide in
excellent yield (94%) and with complete stereoselectivity. The ano-
With the ABO type I and II antigens 4–9 in hand, we conducted
detailed NMR studies to probe if there are any conformational dif-
ferences between the structures. Unequivocally assigning proton
resonances in deprotected oligosaccharides using traditional 1D
and 2D techniques is difficult due to spectral overlap. To overcome
this problem for the ring protons in 4–9, a gradient-enhanced
chemical shift selective filtering (ge-CSSF) technique was used. In
particular, 1D-ge-CSSF-TOCSY experiments were carried out to
provide coupling constants and to assign chemical shifts.31,32 The
use of this experiment enables the straightforward analysis of
the 1H NMR spectra of each ring individually. In addition to analyz-
ing any differences between the type I and II structures, the mea-
surement of 1D-ge-CSSF-TOCSY enabled complete assignment of
all 1H NMR chemical shifts for 4–9. The main limitation of this ap-
proach is that signal transfer throughout each ring is dependent on
the size of the coupling constants. In the case of both galactose and
fucose, when a ring is irradiated at H1, signal transfer from H3 to
meric proton of the
signal at 5.41 ppm as a doublet with a coupling constant of
3.8 Hz, consistent with stereochemistry. Removal of the pivalo-
a-fucopyranoside of 20 showed an anomeric
a
ate ester was achieved using refluxing lithium methoxide in meth-
anol; again, forcing conditions were needed, similar to those
required to prepare 21.
Given the structural similarities between the type I and II anti-
gens, the deprotection strategies would be expected to be analo-
gous and this was indeed the case. The conversion of the azido
group in 38 to an N-acetyl functionality using thiolacetic acid affor-
ded 39 in good (88%) yield (Scheme 8). Interestingly, in contrast to
the preparation of the type I counterpart 22 (see Scheme 3 above),
no acetylation of the alcohol on 39 was observed. Exhaustive
deprotection was achieved in an analogous manner to the type I
counterpart: Birch reduction of 39 afforded a 75% yield of the H
3
H4 is significantly reduced (due to JH3,H4 <3.5 Hz), and transfer
to H5 and H6 does not occur. In most cases this problem could
be circumvented by irradiation of the H5 or H6 resonance and then
allowing the magnetization to transfer the opposite way around
the spin system.
The chemical shift data from these studies are shown in Table 2,
and the associated coupling constants are shown in Table 3. The
OBn
O
O
Ph
O
Ph
O
O
(a)
(b)
HO
BnO
O
O
O
O
O
BnO
HO
N3
31
N3
N3
32
13
OBn
O
(c)
AcO
BnO
O
N3
33
Scheme 6. Reagents: (a) BnBr, NaH, DMF, 98%; (b) Et3SiH, CF3COOH, 4 Å MS, CH2Cl2 83%; (c) Ac2O, DMAP, pyridine, 85%.

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2311
OBn
O
HO
BnO
O
Ph
Ph
N3
O
32
O
O
O
Ph
OBn
O
(a)
O
OBn
O
O
O
RO
O
BnO
PivO
O
BnO
O
O
O
RO
HO
N3
O
N3
36
34 R = Ac
AcO
(b)
(c)
AcO
35 R = H
O
CCl3
NH
15
NH
O
OBn
CCl3
O
(d)
OBn
OBn
BnO
19
Ph
Ph
O
O
O
O
OBn
O
O
O
O
HO
O
BnO
PivO
O
BnO
(e)
O
O
O
O
N3
N3
O
O
OBn
OBn
OBn
OBn
38
37
BnO
BnO
Scheme 7. Reagents: (a) TMSOTf, 4 Å MS, CH2Cl2; (b) NaOCH3, CH3OH, 78% (two steps); (c) (CH3)3CCOCl, pyridine, 88%; (d) TMSOTf, 4 Å MS, Et2O, 94%; (e) LiOCH3, CH3OH,
83%.
Ph
Ph
O
O
O
O
OBn
O
O
OBn
O
O
(a)
HO
O
BnO
HO
O
BnO
O
O
O
O
NHAc
N3
O
O
OBn
OBn
OBn
OBn
38
39
BnO
BnO
(b)
HO
HO
OH
OH
OH
OH
O
O
O
(c)
O
HO
O
HO
HO
O
HO
O(CH2)7CH3
O
O
O
NHAc
NHAc
O
O
OH
OH
7
40
OH
OH
HO
HO
Scheme 8. Reagents: (a) CH3COSH, pyridine, 88%; (b) Na, NH3, CH3OH, THF, 75%; (c) H2, 10% Pd–C, CH3OH, 96%.
residue assignments are shown in Figure 3. In all cases only the
coupling constants for the ring protons were analysed. A signal is
referred to as a multiplet only in instances where the coupling con-
stant could not be determined, either due to second-order coupling
or where inadequate signal transfer in the 1D-ge-CSSF-TOCSY (out-
lined above) resulted in individual proton resonances not being
able to be identified. In cases where insufficient information was
obtained using 1D-ge-CSSF-TOCSY, additional techniques such as
1H–1H COSY, HSQC and HMBC were employed to aid in the identi-
fication of proton resonances. Not surprisingly, the NMR spectra of
all six structures showed significant similarity; little deviation was
observed between the spectra of any of the compounds with re-
spect to both chemical shift (Table 2) and coupling constants
(Table 3).
2.4. Conclusions
In summary, we report the linear syntheses of the A, B and H
type I and II histo-blood group antigens (4–9) attached to either
a 7-octen-1-yl or an octyl aglycone. Key to the synthesis was the

2312
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
Ph
O
O
Ph
OBn
O
O
O
BnO
BnO
HO
O
BnO
OBn
O
O
O
O
N3
(a)
OBn
O
O
OBn
BnO
O
O
O
OBn
O
38
O
BnO
BnO
N3
O
OBn
41
BnO
OBn
O
OBn
BnO
O
CCl3
BnO
OBn
NH
28
(b)
Ph
O
BnO
BnO
HO
OBn
O
OH
O
O
OH
OH
OBn
HO
O
OH
O
O
(c)
BnO
HO
O
O
O
O
O
HO
O
O
O
BnO
O
NHAc
NHAc
O
O
OBn
OH
OBn
OH
42
8
BnO
HO
Scheme 9. Reagents: (a) TMSOTf, 4 Å MS, Et2O, 75%; (b) CH3COSH, pyridine, 91%; (c) Na, NH3, CH3OH, THF, 96%.
preparation of the two reducing-end disaccharides 13 and 32,
which served as the core upon which the rest of the structures
were built. In addition, approaches developed in the synthesis of
the type V and VI antigen synthesis9 were adapted to the synthesis
of the type I and II antigens. Use of these methods enabled the tar-
gets to be obtained with a high level of efficiency. Most notably, the
performed on Silica Gel 60 (40–60
beaded silica gel 6RS-8060, which is manufactured by Iatron Labo-
ratories (Tokyo). C18 silica gel (35–70 m) was manufactured by
lm). Iatrobeads refer to a
l
Toronto Research Chemicals. Optical rotations were measured at
22 2 °C and are in units of deg dmꢀ1 cm3 gꢀ1; in all cases the
concentrations are in the units g/100 mL. 1H NMR spectra were re-
corded at 400 and 500 MHz, and chemical shifts were referenced to
the peak for TMS (0.0 ppm, CDCl3) or CD3OD (3.30 ppm, CD3OD).
For final compounds, analysis was conducted using 1D-ge-CSSF-
TOCSY31,32 at 600 MHz, to identify individual proton resonances.
Samples for 1D-ge-CSSF-TOCSY experiments were prepared in
CD3OD at a concentration of 21 mg/mL, and the experiments were
carried out at 300 K. 13C NMR (APT) spectra were recorded at 125
or 100 MHz, and 13C chemical shifts were referenced to the peak
for internal CDCl3 (77.1 ppm, CDCl3) or CD3OD (49.0, CD3OD). All
spectra were recorded in CDCl3 unless specified otherwise. Melting
points were measured using a Perkin–Elmer Thermal Analysis.
Electrospray-ionization mass spectra were recorded on samples
suspended in mixtures of THF with CH3OH and added NaCl.
preparation of the 7-octen-1-yl b-
ciently prepared without anchimeric assistance. In addition, the
installation of the -fucopyranosyl linkage was entirely stereose-
D-glucopyranoside 11 was effi-
a
-L
lective, taking advantage of the excellent stereoselectivity of the
trichloroacetimidate methodology.29 An NMR study was con-
ducted on each of the type I and II ABO antigens (4–9). Based upon
the chemical shift and coupling constant data, it was clear that the
reducing-end residue has little to no impact on the conformation of
the canonical di- and trisaccharide blood group antigens at the
non-reducing end of the molecule.
3. Experimental
3.1. General methods
3.2. 7-Octen-1-yl 2-acetamido-2-deoxy-3-O-(2-O-(
fucopyranosyl)-3-O-( -galactopyranosyl)-b- -galactopyr-
anosyl)-b- -glucopyranoside (4)
a-L-
a
-D
D
All reagents were purchased from commercial sources and were
used without further purification, unless otherwise stated. Reac-
tion solvents were purchased and were used without purification;
dry solvents were purified by successive passage through columns
of alumina and copper under nitrogen. All reactions were carried
out at room temperature under a positive pressure of argon, unless
otherwise stated. Thin layer chromatography (TLC) was performed
on E. Merck Silica Gel 60 F254 aluminium-backed plates that were
stained by heating (>200 °C) with either p-anisaldehyde in 5% sul-
furic acid in EtOH or 10% ammonium molybdate in 10% sulfuric
acid. Unless otherwise indicated, all column chromatography was
D
Redistilled liquid ammonia (15 mL) was collected in a flask
cooled to ꢀ78 °C and treated with sodium until the blue colour per-
sisted. A solution of tetrasaccharide 30 (67 mg, 0.042 mmol) in THF
(4 mL) and CH3OH (15 lL, 0.37 mmol) was added dropwise, and the
solution was stirred (ꢀ78 °C, 1 h). The reaction was then quenched
by the addition of CH3OH (4 mL), and ammonia was evaporated.
The solution was taken up in CH3OH (100 mL), neutralized with
Amberlite IR 120 (H+) and filtered, and the residue was subjected

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2313
Ph
O
O
Ph
OBn
O
O
O
AcO
AcO
OAc
O
HO
O
BnO
O
O
O
N3
OBn
O
O
(a)
O
N3
OBn
O
O
BnO
38
O
OBn
O
BnO
N3
O
OBn
OBn
43
AcO
OAc
O
BnO
O
CCl3
AcO
N3
NH
(b)
24
Ph
O
HO
O
RO
OH
O
OR
O
OH
OH
HO
OH
O
O
RO
OBn
O
O
O
(d)
AcHN
AcHN
O
O
HO
O
O
BnO
O
O
O
NHAc
NHAc
O
O
OH
OBn
OH
OBn
HO
BnO
9
R = Ac 44
R = H 45
(c)
Scheme 10. Reagents: (a) TMSOTf, 4 Å MS, Et2O, 94%; (b) CH3COSH, pyridine, 76%; (c) NaOCH3, CH3OH, 92%; (d) Na, NH3, CH3OH, THF, 96%.
to C18 chromatography (CH3OH–H2O 1:1) to afford the fully depro-
tected tetrasaccharide 4 (31 mg, 94%) as a colourless oil. [ ] +69.0
68.50, 67.5, 63.8 (C20, C200, C3, C30, C300, C3000, C4, C40, C400, C4000, C5,
C50, C500, C5000), 71.5 (CH@CH2(CH2)5CH2O), 62.3, 62.1, 61.6 (C6, C60,
C6000), 55.6 (C2), 50.5 (C2000), 34.0 (CH@CH2(CH2)5CH2O), 29.4
(CH@CH2(CH2)5CH2O), 29.0 (CH@CH2(CH2)5CH2O), 28.8 (CH@CH2
(CH2)5CH2O), 25.8 (CH@CH2(CH2)5CH2O), 23.2 (CH3C@O), 22.8
(CH3C@O), 16.1 (C600). ESIMS: m/z Calcd [C36H62N2O20]Na+:
865.3788. Found: 865.3788.
a
(c 1.3, MeOH); 1H NMR: see Tables 2 and 3; 13C NMR (175 MHz):
dC 173.2 (C@O), 140.0 (CH2@CH), 114.7 (CH2@CH), 103.6 (C1),
102.1 (C10), 100.5 (C100), 96.2 (C1000), 80.2, 79.9, 77.6, 76.2, 74.5,
73.6, 73.2, 71.4, 71.2 (2C), 70.6, 70.0, 69.8 (C20, C200, C2000, C3, C30,
C300, C3000, C4, C400, C4000, C5, C50, C5000), 70.7 (CH@CH2(CH2)5CH2O),
67.4 (C500), 65.7 (C40), 63.4, 62.7, 62.5 (C6, C60, C6000), 55.9 (C2), 34.8
(CH@CH2(CH2)5CH2O), 30.6 (CH@CH2(CH2)5CH2O), 30.1 (CH@CH2
(CH2)5CH2O), 29.9 (CH@CH2(CH2)5CH2O), 26.9 (CH@CH2(CH2)5
CH2O), 23.2 (CH3C@O), 16.5 (C600). ESIMS: m/z Calcd [C34H59NO20]-
Na+: 824.3523. Found: 824.3522.
3.4. 7-Octen-1-yl 2-acetamido-2-deoxy-3-O-(2-O-(
a-L-
fucopyranosyl)-b- -galactopyranosyl)-b- -glucopyranoside (6)
D
D
Redistilled liquid ammonia (15 mL) was collected in a flask
cooled to ꢀ78 °C and treated with sodium until the blue colour
persisted. A solution of trisaccharide 22 (130 mg, 0.120 mmol) in
3.3. 7-Octen-1-yl 2-acetamido-3-O-(3-O-(2-acetamido-2-deoxy-
a-
D
-galactopyranosyl)-2-O-(
a-
L
-fucopyranosyl)-b-
D-
THF (4 mL) and CH3OH (24 lL, 0.60 mmol) was added dropwise,
galactopyranosyl)-2-deoxy-b-
D-glucopyranoside (5)
and the solution was stirred (ꢀ78 °C, 2 h). The reaction was then
quenched by the addition of CH3OH (4 mL), and ammonia was
evaporated. The solution was taken up in CH3OH (100 mL),
neutralized with Amberlite IR 120 (H+) and filtered, and the residue
was subjected to C18 chromatography (CH3OH–H2O 1:1) to afford
the fully deprotected trisaccharide 6 as a colourless oil (70 mg,
Redistilled liquid ammonia (20 mL) was collected in a flask
cooled to ꢀ78 °C and treated with sodium until the blue colour per-
sisted. A solution of tetrasaccharide 27 (58 mg, 0.045 mmol) in THF
(4 mL) and CH3OH (9.1 lL, 0.225 mmol) was added dropwise, and
the solution was stirred (ꢀ78 °C, 1 h). The reaction was then
quenched by the addition of CH3OH (4 mL), and ammonia was
evaporated. The solution was taken up in CH3OH (100 mL), neutral-
ized with Amberlite IR 120 (H+), and filtered, and the residue was
subjected to C18 chromatography (CH3OH–H2O 1:1) to afford the
fully deprotected tetrasaccharide 5 (33.5 mg, 88%) as a colourless
92%). [
a
] ꢀ62.7 (c 1.0, CH3OH); 1H NMR: see Tables 2 and 3; 13C
NMR (125 MHz, CD3OD): dC 173.1 (C@O), 140.1 (CH2@CH), 114.8
(CH2@CH), 103.6 (C1), 102.2 (C10) 101.2 (C100), 80.3, 78.0, 77.7,
77.0, 75.6, 73.6, 71.4, 70.8, 70.5, 70.3, 67.6 (C20, C200, C3, C30, C300,
C4, C40, C400, C5, C50, C500), 70.7 (CH@CH2(CH2)5CH2O), 62.7, 62.6
(C6, C60), 56.1 (C2), 34.9 (CH@CH2(CH2)5CH2O), 30.6 (CH@CH2
(CH2)5CH2O), 30.2 (CH@CH2(CH2)5CH2O), 30.0 (CH@CH2(CH2)5
CH2O), 27.0 (CH@CH2(CH2)5CH2O), 23.2 (CH3C@O), 16.6 (C600).
ESIMS: m/z Calcd [C28H49NO15]Na+: 662.2994. Found: 662.3000.
oil. [a
] +27.1 (c 0.2, H2O); 1H NMR: see Tables 2 and 3; 13C NMR
(125 MHz, D2O): dC 175.7 (C@O), 174.5 (C@O), 141.2 (CH2@CH),
114.9 (CH2@CH), 102.8, 100.8, 100.0 (C1, C10, C100), 92.1 (C1000),
78.3, 76.33, 76.27, 75.7, 74.7, 72.7, 71.8, 70.6, 69.7, 69.4, 68.53,

2314
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
Table 2
1H Assignments (ppm) of the ABH type I–II antigens 4–9, recorded at 600 MHz with CD3OD as solvent
Proton
A Type 1 5
A Type II 9
B Type 1 4
B Type II 8
H Type 1 6
H Type II 7
H1000
H2000
H3000
H4000
H5000
H6000
H6000
5.15 (d)
5.15 (d)
5.14 (d)
5.15 (d)
3.84 (dd)
3.78 (dd)
3.9 (d)
4.14–4.10 (m)
3.76–3.71 (m)
3.70–3.66 (m)
4.29 (dd)
3.85 (dd)
3.87 (d)
4.20 (dd)
3.75 (dd)
3.68 (dd)
4.32 (dd)
3.82 (dd)
3.89 (d)
4.16 (dd)
3.76 (dd)
3.68 (dd)
3.83 (dd)
3.81 (dd)
3.89 (dd)
4.16 (dd)
3.73 (dd)
3.68 (dd)
H100
H200
H300
H400
H500
H600
5.23 (s)
3.69 (dd)
3.65 (dd)
3.72 (d)
4.36 (q)
1.19 (d)
S.33 (d)
3.72 (dd)
3.68 (dd)
3.69 (d)
4.32 (q)
1.21 (d)
5.21 (d)
3.7 (dd)
3.65 (dd)
3.71 (d)
4.45 (q)
1.18 (d)
5.32 (m)
3.73 (m)
3.69–3.66 (m)
3.69–3.66 (m)
4.29 (q)
5.20 (d)
5.21 (d)
3.73 (dd)
3.68 (dd)
3.71–3.69 (m)
4.29 (q)
3.77–3.71 (m)
3.77–3.71 (m)
3.66(d)
4.17 (q)
1.20 (d)
1.20 (d)
1.19 (d)
H10
H20
H30
H40
H50
H60
H60
4.57 (d)
3.91 (dd)
3.86 (dd)
4.09 (d)
3.49 (dd)
3.77–3.73 (m)
3.66–3.65 (m)
4.52 (d)
3.99 (dd)
3.90 (dd)
4.09 (d)
3.29–3.25 (m)
3.81–3.77 (m)
3.72–3.66 (m)
4.60–4.56 (m)
3.93–3.87 (m)
3.93–3.87 (m)
4.10 (s)
3.54 (dd)
3.79 (dd)
4.53 (d)
4.54–4.51 (m)
3.71–3.65 (m)
3.71–3.65 (m)
3.80–3.78 (m)
3.52 (dd)
4.50–4.44 (m)
3.75–3.68 (m)
3.75–3.68 (m)
3.78(d)
3.54 (dd)
3.65–3.61 (m)
3.75–3.73 (m)
3.98 (dd)
3.94 (dd)
4.12 (d)
3.60–3.54 (m)
3.79 (dd)
3.67 (dd)
3.67–3.64 (m)
3.77–3.74 (m)
3.68 (dd)
H1
H2
H3
H4
H5
H6
H6
4.25–4.20 (m)
3.86–3.79 (m)
3.86–3.79 (m)
3.41–3.37 (m)
3.32–3.28 (m)
3.78–3.74 (m)
3.69–3.66 (m)
4.39 (d)
3.71 (dd)
3.69 (dd)
3.59 (dd)
3.28 (dd)
3.90 (d)
3.78(d)
4.25–4.19 (m)
3.86–3.77 (m)
3.86–3.77 (m)
3.41–3.37 (m)
3.31–3.28 (m)
3.93–3.83 (m)
3.76–3.64 (m)
4.36 (d)
4.28–4.22 (m)
3.84–3.78 (m)
3.84–3.78 (m)
3.42–3.38 (m)
3.32–3.28 (m)
3.74–3.65 (m)
3.92–3.84 (m)
4.37 (d)
3.70 (dd)
3.59 (dd)
3.66 (dd)
3.30–3.26 (m)
3.90 (dd)
3.77 (dd)
3.72 (dd)
3.59 (dd)
3.69 (dd)
3.33–3.29 (m)
3.89 (dd)
3.83 (dd)
Other
CH2O
CH2O
3.91–3.87 (m)
3.44–3.40 (m)
4.99–4.95 (m)
4.91–4.89 (m)
5.84–5.76 (dddd)
2.08–2.01 (m)
1.57–1.47 (m)
1.42–1.26 (m)
1.99 (s)
3.86–3.83 (m)
3.45–3.41 (dt)
5.00–4.94 (m)
4.92–4.88 (m)
5.84–5.76 (dddd)
2.07–2.01 (m)
1.58–1.48 (m)
1.42–1.27 (m)
2.00 (s)
3.92–3.83 (m)
3.46–3.37 (m)
4.99–4.95 (m)
4.92–4.89 (m)
5.84–5.75 (dddd)
2.07–2.01 (m)
1.57–1.46 (m)
1.42–1.27 (m)
1.99 (s)
3.91–3.83 (m)
3.48–3.43 (dt)
5.00–4.95 (m)
4.93–4.88 (m)
5.84–5.76 (dddd)
2.08–2.01 (m)
1.58–1.49 (m)
1.43–1.25 (m)
1.95 (s)
3.45–3.37 (m)
3.91–3.83 (m)
4.99–4.93 (m)
4.92–4.88 (m)
5.84–5.75 (dddd)
2.07–2.00 (m)
1.58–1.46 (m)
1.41–1.26 (m)
1.97 (s)
3.47–3.42 (m)
3.91–3.82 (dt)
4.99–4.94 (m)
4.92–4.88 (m)
5.83–5.75 (dddd)
2.07–2.01 (m)
1.58–1.48 (m)
1.41–1.28 (m)
1.96 (s)
CH@CH2
CH@CH2
CH@CH2
(CH2)5
(CH2)5
(CH2)5
CH3CO
CH3CO
1.99 (s)
1.96 (s)
Table 3
2JH,H,3JH,H (Hz) of the ABH type I–II antigens 4–9, recorded at 600 MHz with CD3OD as solvent
Coupling
A Type I 5
A Type II 9
B Type I 4
B Type II 8
H Type I 6
H Type II 7
3JH-l
3.8
10.8
3.4
0
7.6
4.1
11.7
3.7
11.0
3.0
0
7.2
4.5
11.5
3.5
10.2
3.1
0.8
7.3
3.8
10.1
2.9
m
rr
m
000 ,H-2000
3JH-2
000 ,H-3000
3JH-3
000 ,H-4000
3JH-4
000 ,H-5000
3JH-5
000 ,H-6a000
3JH-5
4.3
11.4
000 ,H-6b
2JH-6a
m
000 ,H-6b000
3 JH-l
3.9
10.5
3.1
3.9
9.5
3.3
6.6
4.1
9.3
3.3
6,6
3.9
9.9
m
3.8
10.0
3.1
6.6
3.3
m
2.8
6.6
00 ,H-200
3JH-2
00 ,H-300
3JH-3
00 ,H-400
3JH-5
6.5
6.6
00 ,H-600
3
0
0
JH-l ,H-2
7.3
9.9
3.1
7.9
4.3
m
7.7
9.8
3.0
rr
m
rr
m
m
0
7.8
4.4
11.4
7.4
9.7
2.7
7.3
3.9
11.6
m
m
m
7.7
4.3
m
3
3
3
3
2
0
0
0
JH-2 ,H-3
JH-3 ,H-4
JH-5 ,H-6a
JH-5 ,H-6b
0
4.0
7.6
4.0
0
0
0
0
0
0
JH-6a ,H-6b
3JH-l,H-2
m
m
m
m
m
m
m
8.4
9.3
8.9
9.8
4.9
0
m
m
m
m
m
m
m
8.4
10.4
8.8
9.2
5.0
m
m
m
m
m
m
rr
8.4
10.5
8.8
10.4
4.4
1.9
3JH-2,H-3
3JH-3,H-4
3JH-4,H-5
3JH-5,H-6a
3JH-5,H-6b
2JH-6a,H-6b
1.8
11.9
11.2
12.0
CH2O
m
9.7, 6.6
m
9.7, 6.6
m
9.6, 6.6
CH@CH2
17.1, 10.2, 6.7, 6.7
17.0, 10.2, 6.7, 6,7
17.0, 10.2, 6.7, 6.7
17.0, 10.2, 6.7, 6.7
17.0, 10.2, 6.7, 6.7
17.0, 10.2, 6.7, 6.7

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2315
HO
in CH3OH, and the solution was stirred for 2 h. The solution was
then neutralized with Amberlite IR 120 (H+) and filtered, and the
residue was subjected to flash chromatography (CH2Cl2–CH3OH
10:1) to afford triol 45 (440 mg, 92%) as a colourless oil. Redistilled
liquid ammonia (20 mL) was collected in a flask cooled to (ꢀ78 °C)
and treated with sodium until the blue colour persisted. A solution
of tetrasaccharide 45 (221 mg, 0.160 mmol) in THF (4 mL) and
OH
O
OH
OH
HO
OH
O
O
HO
O
O
HO
O
H1
H1′′′
O
H1′
NHAc
H1′′
OH
O
CH3OH (39 lL, 0.961 mmol) was added dropwise, and the mixture
8
OH
HO
B Type II
was stirred (ꢀ78 °C, 1 h). The reaction was then quenched by the
addition of CH3OH (4 mL), and ammonia was evaporated. The solu-
tion was taken up in CH3OH (100 mL), neutralized with Amberlite
IR 120 (H+) and filtered, and the residue was subjected to C18 chro-
matography (CH3OH–H2O 1:1) to afford the fully deprotected tet-
Figure 3. Nomenclature system used for 4–9, using the B type II antigen 8 for illus-
trative purposes. This nomenclature is used for the data presented in Tables 2 and 3.
3.5. 7-Octen-1-yl 2-acetamido-2-deoxy-4-O-(2-O-(
a-L-
fucopyranosyl)-b- -galactopyranosyl)-b- -glucopyranoside (7)
D
D
rasaccharide 9 (129 mg, 96%) as a colourless oil. [a] +5.8 (c 1.6,
CH3OH); 1H NMR: see Tables 2 and 3; 13C NMR (125.7 MHz,
CD3OD): dC 174.4 (C@O), 173.3 (C@O), 140.0 (CH2@CH), 114.8
(CH2@CH), 104.2 (C10), 100.5 (C100), 98.1 (C1), 93.9 (C1000), 77.6,
77.4, 76.2, 75.8, 73.4, 72.7, 71.8, 71.6, 70.6, 70.1, 70.0, 69.88 (C20,
C200, C3, C30, C300, C3000, C4, C400, C4000, C5, C50, C5000), 68.94
(CH@CH2(CH2)5CH2O), 68.4 (C500), 64.9 (C40), 63.4, 62.8, 62.5 (C6,
C60, C6000), 51.4, 50.9 (C2, C2000), 34.9 (CH@CH2(CH2)5CH2O), 30.6
Redistilled liquid ammonia (20 mL) was collected in a flask
cooled to (ꢀ78 °C) and treated with sodium until the blue colour
persisted. A solution of trisaccharide 39 (185 mg, 0.157 mmol) in
THF (4 mL) and CH3OH (38 lL, 0.94 mmol) was added dropwise,
and the mixture was stirred (ꢀ78 °C, 1 h). The reaction was then
quenched by the addition of CH3OH (4 mL), and ammonia was evap-
orated. The solution was taken up in CH3OH (100 mL), neutralized
with Amberlite IR 120 (H+) and filtered, and the residue was sub-
jected to C18 chromatography (CH3OH–H2O 1:1) to afford the fully
(CH@CH2(CH2)5CH2O),
30.1
(CH@CH2(CH2)5CH2O),
30.0
(CH@CH2(CH2)5CH2O), 27.2 (CH@CH2(CH2)5CH2O), 22.9 (CH3CO),
22.8 (CH3CO), 16.9 (C600). ESIMS: m/z Calcd [C36H62NO20]Na+:
865.3788. Found: 865.3783.
deprotected trisaccharide 7 (75 mg, 75%) as a colourless oil. [a]
ꢀ78.9 (c 0.4, CH3OH); 1H NMR: see Tables 2 and 3; 13C NMR
(125.7 MHz, CD3OD): dC 178.4 (C@O), 140.1 (CH2@CH), 114.8
(CH2@CH), 102.9, 102.6 (C1, C10), 101.9 (C100), 79.1, 78.3, 77.1, 77.0,
75.3, 74.1, 73.6, 71.7, 70.8, 70.7 (C20, C200, C3, C30, C300, C4, C40, C400,
C5, C50), 70.7 (CH@CH2(CH2)5CH2O), 68.3 (C500), 62.6, 61.7 (C6, C60),
56.8 (C2), 34.8 (CH@CH2(CH2)5CH2O), 30.6 (CH@CH2(CH2)5CH2O),
30.1 (CH@CH2(CH2)5CH2O), 30.0 (CH@CH2(CH2)5CH2O), 27.0
(CH@CH2(CH2)5CH2O), 23.0 (CH3CO), 16.8 (C600). ESIMS: m/z Calcd
[C28H49NO15]Na+: 662.2994. Found: 662.2985.
3.8. 7-Octen-1-yl 2-azido-4,6-O-benzylidene-2-deoxy-b-
glucopyranoside (13) and 7-Octen-1-yl 2-Azido-4,6-O-
D-
benzylidene-2-deoxy-a-D-glucopyranoside (14)
A
stirred solution of trichloroacetimidate27 10 (8.69 g,
18.3 mmol) and 7-octen-1-ol (2.82 g, 22.0 mmol) in dry CH2Cl2
(50 mL) was treated with 4 Å molecular sieves (3.5 g), and the
mixture was stirred (rt, 1 h). The mixture was cooled (ꢀ30 °C),
treated with TMSOTf (300 lL) and allowed to slowly warm to
3.6. 7-Octen-1-yl 2-acetamido-2-deoxy-4-O-(2-O-(
fucopyranosyl)-3-O-( -galactopyranosyl)-b- -galactopyr-
anosyl)-b- -glucopyranoside (8)
a
-
L
-
0 °C. The mixture was neutralized by the addition of Et3N
(1 mL), filtered, concentrated and subjected to flash chromatogra-
phy (EtOAc–hexanes 1:3) to give an inseparable a/b mixture 11,
a-D
D
D
which was used immediately in the subsequent step. The oil
was taken up in CH3OH (80 mL) and treated with a catalytic
amount of NaOCH3 in CH3OH, and the solution was stirred (rt,
1 h). The solution was then neutralized with Amberlite IR 120
(H+), and the mixture was filtered; concentration followed by
flash chromatography (EtOAc–hexanes 2:1) yielded triol 12
Redistilled liquid ammonia (20 mL) was collected in a flask
cooled to (ꢀ78 °C) and treated with sodium until the blue colour
persisted. A solution of tetrasaccharide 42 (244 mg, 0.144 mmol)
in THF (4 mL) and CH3OH (58 lL, 1.44 mmol) was added dropwise,
and the mixture was stirred (ꢀ78 °C, 1 h). The reaction was then
quenched by the addition of CH3OH (4 mL), and ammonia was evap-
orated. The solution was taken up in CH3OH (100 mL), neutralized
with Amberlite IR 120 (H+) and filtered, and the residue was sub-
jected to C18 chromatography (CH3OH–H2O 1:1) to afford the fully
(3.32 g) as an inseparable
a/b mixture. A solution of triol 12
(3.32 g, 10.5 mmol) in dry DMF (20 mL) was then treated with
benzaldehyde dimethyl acetal (2.13 g, 14.0 mmol) and p-TsOH
(100 mg), and the solution was stirred (50 °C, 4 h). The solution
was treated with Et3N (1 mL) and concentrated, and the residue
was subjected to flash chromatography (EtOAc–hexanes 1:3) to
firstly afford the b glycoside 13 as a colourless oil (3.05 g, 41%).
deprotected tetrasaccharide 8 (111 mg, 96%) as a colourless oil. [a]
ꢀ28.9 (c 0.5, CH3OH); 1H NMR: see Tables 2 and 3; 13C NMR
(125.7 MHz, CD3OD): dC 173.4 (C@O), 140.1 (CH2@CH), 114.8
(CH2@CH), 102.8, 102.2 (C1, C10), 100.3 (C100), 96.1 (C1000), 79.8,
78.6, 77.1, 76.7, 74.1, 73.8, 73.6, 73.1, 71.8, 71.4, 71.3, 70.0, 69.9,
67.6, 65.8 (C20, C200, C2000, C3, C30, C300, C3000, C4, C40, C400, C4000, C5, C50,
C500, C5000), 70.7 (CH@CH2(CH2)5CH2O), 63.3, 62.6, 61.8 (C6, C60,
C6000), 56.8 (C2), 34.8 (CH@CH2(CH2)5CH2O), 30.6 (CH@CH2(CH2)5
CH2O), 30.1 (CH@CH2(CH2)5CH2O), 30.0 (CH@CH2(CH2)5CH2O),
27.0 (CH@CH2(CH2)5CH2O), 23.0 (CH3CO), 16.6 (C600). ESIMS: m/z
Calcd [C34H59NO20]Na+: 824.3523. Found: 824.3518.
[
a
] ꢀ38.4 (c 0.4, CH2Cl2); Rf 0.18 (EtOAc–hexanes 7:3); 1H NMR
(500 MHz): dH 7.52–7.35 (5H, m, Ph), 5.87–5.76 (1H, m, CH@CH2),
5.54 (1H, s, PhCH), 5.04–4.92 (2H, m, CH@CH2), 4.42 (1H, d,
J1,2 = 8.0 Hz, H1), 4.34 (1H, dd, J6,6 = 10.3 Hz, J5,6 = 5.0 Hz, H6),
3.97–3.89 (1H, m, CH@CH2(CH2)5CH2O), 3.79 (1H, dd,
J6,6 = J5,6 = 10.3 Hz,
H6),
3.69–3.51
(3H,
m,
H3,
H4,
CH@CH2(CH2)5CH2O), 3.45–3.35 (2H, m, H2, H5), 2.69 (1H, br s,
OH), 2.10–1.99 (2H, m, CH@CH2(CH2)5CH2O), 1.73–1.56 (2H, m,
CH@CH2(CH2)5CH2O), 1.46–1.25 (6H, m, CH@CH2(CH2)5CH2O).
13C NMR (125 MHz): dC 139.0 (CH@CH2), 136.8 (Ph), 129.4 (Ph),
128.4 (Ph), 126.2 (Ph), 114.3 (CH@CH2), 102.7, 102.0 (PhCH, C1),
80.6, 72.0, 66.5, 66.2 (C2, C3, C4, C5), 70.7 (CH@CH2(CH2)5CH2O),
68.5 (C6), 33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O),
28.81 (CH@CH2(CH2)5CH2O), 28.78 (CH@CH2(CH2)5CH2O), 25.8
(CH@CH2(CH2)5CH2O). ESIMS: m/z Calcd [C21H29N3O5]Na+:
3.7. 7-Octen-1-yl 2-acetamido-4-O-(3-O-(2-acetamido-2-deoxy-
a-
D
-galactopyranosyl)-2-O-(
a-
L
-fucopyranosyl)-b-
D-
galactopyranosyl)-2-deoxy-b-
D-glucopyranoside (9)
A solution of tetrasaccharide 44 (520 mg, 0.345 mmol) in
CH3OH (20 mL) was treated with a catalytic amount of NaOCH3

2316
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
426.2000. Found: 426.2002. Further elution yielded
a
glycoside
3.10. 7-Octen-1-yl 2-azido-3-O-(4,6-O-benzylidene-3-O-
pivaloyl-2-O-(2,3,4-tri-O-benzyl- -fucopyranosyl)-b-
galactopyranosyl)-2-deoxy-b- -glucopyranoside (20)
14 as a colourless oil (0.8 g, 9%). [ ] +80.3 (c 0.7, CH2Cl2); Rf
a
a-L
D-
0.16 (EtOAc–hexanes 7:3); 1H NMR (500 MHz): dH 7.41–7.37
(5H, m, Ph), 5.87–5.77 (1H, m, CH@CH2), 5.55 (s, PhCH), 5.04–
4.93 (2H, m, CH@CH2), 4.90 (1H, d, J1,2 = 3.6 Hz, H1), 4.28 (1H,
dd, J6,6 = 10.2 Hz, J5,6 = 4.9 Hz, H6), 4.23 (1H, dd, J2,3 = 10.0 Hz,
J3,4 = 9.6 Hz, H3), 3.87 (1H, ddd, J5,6 = 9.9 Hz, J4,5 = 9.9 Hz,
J5,6 = 4.9 Hz, H5), 3.77–3.71 (2H, m, H6, CH@CH2(CH2)5CH2O),
3.54–3.46 (2H, m, H4, CH@CH2(CH2)5CH2O), 3.23 (1H, dd,
J2,3 = 10.0 Hz, J1,2 = 3.6 Hz, H2), 2.76 (1H, br s, OH), 2.09–2.03
(2H, m, CH@CH2(CH2)5CH2O), 1.70–1.62 (2H, m, CH@CH2(CH2)5
CH2O), 1.46–1.33 (6H, m, CH@CH2(CH2)5CH2O). 13C NMR
(125 MHz): dC 139.0 (CH@CH2), 136.9 (Ph), 129.5 (Ph), 128.4
(Ph), 126.4 (Ph), 114.3 (CH@CH2), 102.1 (PhCH), 98.6 (C1), 82.0
(C4), 68.9, 68.8 (C6, CH@CH2(CH2)5CH2O), 68.7 (C3), 63.1, 62.4
(C2, C5), 33.7 (CH@CH2(CH2)5CH2O), 29.4 (CH@CH2(CH2)5CH2O),
D
A solution of acceptor 18 (415 mg, 0.563 mmol) in dry Et2O–
CH2Cl2 (90:10, 20 mL) was stirred over 4 Å molecular sieves (rt,
1 h). The solution was then cooled (ꢀ10 °C), treated with TMSOTf
(20 l
L), followed by dropwise addition of trichloroacetimidate29
19 (1.02 g, 13.8 mmol) in dry Et2O (15 mL). The mixture was trea-
ted with Et3N (0.5 mL) and filtered, and the resulting residue was
subjected to flash chromatography (EtOAc–hexanes 1:3) to yield
trisaccharide 20 as a colourless oil (510 mg, 80%). [
a
] ꢀ20.7 (c
0.2, CH2Cl2); Rf 0.59 (EtOAc–hexanes 3:7); 1H NMR (500 MHz): dH
7.55–7.22 (25H, m, Ph), 5.87–5.77 (1H, m, CH2@CH), 5.41 (1H, d,
J1 ,2 = 1.5 Hz, H100), 5.48 (1H, s, PhCH), 5.37 (1H, s, PhCH), 5.04–
00 00
4.92 (4H, m, H30, PhCH2, CH2@CH), 4.79, 4.74 (2H, AB, J = 11.5 Hz,
PhCH2), 4.76 (1H, d, J1 ,2 = 8.1 Hz, H10), 4.69 (1H,
A of AB,
0
0
28.81
(CH@CH2(CH2)5CH2O),
28.78
(CH@CH2(CH2)5CH2O),
25.9 (CH@CH2(CH2)5CH2O). ESIMS: m/z Calcd [C21H29N3O5]Na+:
J = 11.7 Hz, PhCH2), 4.79, 4.63 (2H, AB, J = 11.5 Hz, PhCH2), 4.51
(1H, q, J5 ,6 = 6.3 Hz, H500), 4.42 (1H, d, J1,2 = 7.7 Hz, H1), 4.34–
00 00
426.2000. Found: 426.1995.
4.29 (2H, m, H6, H60), 4.24 (1H, d, J1 ,2 = J2 ,3 = 8.5, H20), 4.13–4.07
(3H, m, H200, H300, H400), 3.97–3.91 (1H, m, CH@CH2(CH2)5CH2O),
3.83–3.70 (5H, m, H3, H4, H40, H6, H60), 3.63–3.56 (1H, m,
CH@CH2(CH2)5CH2O), 3.43–3.35 (2H, m, H2, H5), 3.04–2.99 (1H,
m, H50), 2.11–2.03 (2H, m, CH@CH2(CH2)5CH2O), 1.73–1.63 (2H,
m, CH@CH2(CH2)5CH2O), 1.46–1.31 (6H, m, CH@CH2(CH2)5CH2O),
0
0
0
0
3.9. 7-Octen-1-yl 2-azido-3-O-(4,6-O-benzylidene-3-O-pivaloyl-
b-D
-galactopyranosyl)-2-deoxy-b-D-glucopyranoside (18)
A solution of acceptor 13 (2.3 g, 5.7 mmol) in dry CH2Cl2
1.20 (3H, d, J5 ,6 = 6.3 Hz, H600), 1.08 (9H, s, (CH3)3C). 13C NMR
(20 mL) was stirred over 4 Å molecular sieves (1.5 g), and the mix-
ture was stirred (rt, 1 h). The solution was then cooled (ꢀ40 °C)
and treated with TMSOTf (0.2 mL), followed by the dropwise addi-
tion of trichloroacetimidate 1533 (8.0 g, 16 mmol). The mixture
was allowed to warm to 0 °C, and it was then neutralized by the
addition of Et3N (2 mL). Concentration and flash chromatography
(EtOAc–hexanes 1:1) of the residue afforded a colourless oil that
was immediately used in the next step. The colourless oil was ta-
ken up in CH3OH (100 mL), treated with a solution of NaOCH3 in
CH3OH and stirred (rt, 3 h). The solution was neutralized with
Amberlite IR 120 (H+), filtered and subjected to flash chromatogra-
phy (EtOAc–hexanes 1:1) to afford diol 17 as a colourless oil
(2.3 g, 63%). A solution of 17 (2.20 g, 3.37 mmol) in dry pyridine
(30 mL) was treated with trimethylacetyl chloride (600 mg,
5 mmol), and the solution was stirred (rt, 30 min). Further tri-
methylacetyl chloride (600 mg, 5 mmol) was added, and the solu-
tion was stirred (rt, 30 min). The solution was then concentrated
and subjected to flash chromatography (EtOAc–hexanes 2:3) to
00 00
(125 MHz): dC 177.9 (C@O), 139.01 (Ph), 138.98 (CH2@CH), 138.6
(Ph), 138.4 (Ph), 137.6 (Ph), 136.8 (Ph), 129.2 (Ph), 128.7 (Ph),
128.5 (Ph), 128.4 (Ph), 128.33 (Ph), 128.28 (Ph), 128.2 (Ph), 128.0
(2C, Ph), 127.6 (Ph), 127.5 (Ph), 127.44 (Ph), 127.38 (Ph), 126.2
(2C, Ph), 114.3 (CH2@CH), 102.8 (C1), 101.05 (C10), 101.7 (PhCH),
100.8 (PhCH), 96.8 (C100), 79.9, 79.7, 78.0, 77.5, 76.55, 76.52 (C3,
C30, C300, C4, C40, C400), 75.0 (PhCH2), 73.5 (PhCH2), 72.9 (PhCH2),
72.6, 70.1 (C20, C200), 70.7 (CH@CH2(CH2)5CH2O), 68.8, 68.6
(C6, C60), 66.5 (C500), 66.4, 65.9, 65.7 (C2, C5, C50), 38.8 ((CH3)3C),
33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O), 28.83
(CH@CH2(CH2)5CH2O),
28.78
(CH@CH2(CH2)5CH2O),
27.0
((CH3)3C), 25.8 (CH@CH2(CH2)5CH2O), 16.9 (C600). ESIMS: m/z Calcd
[C66H79N3O15]Na+: 1176.5403. Found: 1176.5402.
3.11. 7-Octen-1-yl 2-azido-3-O-(4,6-O-benzylidene-2-O-(2,3,4-
tri-O-benzyl-a-L-fucopyranosyl)-b-D-galactopyranosyl)-2-
deoxy-b- -glucopyranoside (21)
D
afford 18 as a colourless oil (2.2 g, 89%). [a] +5.8 (c 0.1, CH2Cl2);
Rf 0.75 (EtOAc–hexanes 2:3); 1H NMR (500 MHz): dH 7.52–7.46
(4H, m, Ph), 7.38–7.30 (6H, m, Ph), 5.86–5.76 (1H, m, CH2@CH),
5.54 (1H, s, PhCH), 5.46 (1H, s, PhCH), 5.04–4.92 (2H, m, CH2@CH),
A solution of pivaloyl ester 20 (1.456 g, 1.26 mmol) in CH3OH
(150 mL) was treated with catalytic LiOCH3 (100 mg), and the
solution was heated at reflux (7 d). The solution was then concen-
trated, extracted with EtOAc (400 mL) and washed with satd aq
NaHCO3 and brine. The organic extract was then dried, concen-
trated and subjected to flash chromatography (EtOAc–hexanes
4.78 (1H, dd, J2 ,3 = 9.5 Hz, J3 ,4 = 3.6 Hz, H30), 4.49 (1H, d,
0
0
0
0
J1 ,2 = 7.9 Hz, H10), 4.47 (1H, d, J1,2 = 8.0 Hz, H1), 4.37–4.29 (2H,
0
0
m, H40, H6), 4.17 (1H, d, J6 ,6 = 12.1 Hz, H60), 4.05 (1H, dd,
0
0
J2 ,3 = 9.5 Hz, J1 ,2 = 8.2 Hz, H20), 3.96–3.88 (2H, m, H60,
CH@CH2(CH2)5CH2O), 3.80 (1H, dd, J6,6 = J5,6 = 10.1 Hz, H6), 3.77–
3.72 (2H, m, H3, H4), 3.62–3.49 (2H, m, H2, CH@CH2(CH2)5CH2O),
3.46–3.35 (1H, m, H5), 3.33–3.29 (1H, m, H50), 3.02–2.96 (1H, br s,
OH), 2.11–2.01 (2H, m, CH@CH2(CH2)5CH2O), 1.72–1.60 (2H, m,
CH@CH2(CH2)5CH2O), 1.46–1.30 (6H, m, CH@CH2(CH2)5CH2O),
1.22 (9H, s, (CH3)3C). 13C NMR (125 MHz): dC 178.3 (C@O), 139.0
(CH2@CH), 137.9 (Ph), 137.0 (Ph), 129.1 (Ph), 128.7 (Ph), 128.2
(Ph), 128.0 (Ph), 126.02 (Ph), 125.96 (Ph), 114.3 (CH2@CH), 104.5
(C10), 102.7 (C1), 101.4 (PhCH), 100.5 (PhCH), 79.9, 79.8 (C3, C4),
73.21, 73.20 (C30, C40), 70.8 (CH@CH2(CH2)5CH2O), 69.1 (C20),
68.8, 68.5 (C6, C60), 67.0 (C50), 66.3 (C5), 65.5 (C2), 39.0
((CH3)3C), 33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O),
28.80 (CH@CH2(CH2)5CH2O), 28.77 (CH@CH2(CH2)5CH2O), 27.1
((CH3)3C), 25.7 (CH@CH2(CH2)5CH2O). ESIMS: m/z Calcd
[C44H59N3O13]Na+: 760.3416. Found: 760.3415.
0
0
0
0
3:7) to afford alcohol 21 as a colourless oil (1.10 g, 82%). [a]
ꢀ20.7 (c 0.1, CH2Cl2); Rf 0.26 (EtOAc–hexanes 3:7); 1H NMR
(500 MHz): dH 7.63–7.51 (4H, m, Ph), 7.42–7.19 (21H, m, Ph),
5.89–5.79 (1H, m, CH2@CH), 5.57 (1H, s, PhCH), 5.54 (1H, s, PhCH),
5.33 (1H, s, H100), 5.06–4.94 (3H, m, PhCH2, CH2@CH), 4.85 (1H, A
of AB, J = 11.5 Hz, PhCH2), 4.84–4.74 (3H, m, PhCH2), 4.68 (1H, d,
´
0
0
J1 ,2 = 7.1 Hz, H1), 4.66 (1H, A of AB, J = 11.1 Hz PhCH2), 4.42 (1H,
d, J1,2 = 8.2 Hz, H1), 4.35 (1H, dd, J6,6 = 10.5 Hz, J5,6 = 4.8 Hz, H6),
4.31 (1H, q, J5 ,6 = 6.3 Hz, H500), 4.23 (1H, d, J6 ,6 = 12.4 Hz, H60),
00 00
0
0
4.18 (1H, d, J3 ,4 = 3.2 Hz, H40), 4.13–4.06 (2H, m, H200, H300),
3.98–3.90 (3H, m, H20, H6, CH@CH2(CH2)5CH2O), 3.86–3.79 (3H,
m, H400, H60, OH), 3.78–3.72 (2H, m, H3, H30), 3.68 (1H, dd,
0
0
J3,4 = 9.0 Hz,
J4,5 = 9.0 Hz,
H4),
3.63–3.58
(1H,
m,
CH@CH2(CH2)5CH2O), 3.44 (1H, dd, J1,2 = 8.2 Hz, J2,3 = 8.2 Hz, H2),
3.41–3.36 (1H, m, H5), 3.26 (1H, s, H50), 2.12–2.04 (2H, m,
CH@CH2(CH2)5CH2O), 1.78–1.56 (2H, m, CH@CH2(CH2)5CH2O),

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2317
00 00
1.49–1.32 (6H, m, CH@CH2(CH2)5CH2O), 1.24 (d, 3H, J5 ,6 = 6.3 Hz,
colourless oil (68 mg, 83%). [
a
] ꢀ15.1 (c 0.1, H2O); 1H NMR
H600). 13C NMR (125 MHz): dC 139.0 (CH2@CH), 138.8 (Ph), 138.7
(Ph), 137.9 (Ph), 137.8 (Ph), 137.1 (Ph), 129.0 (Ph), 128.7 (Ph),
128.41 (Ph), 128.38 (Ph), 128.36 (Ph), 128.23 (2C, Ph), 128.19
(Ph), 128.1 (Ph), 127.8 (Ph), 127.54 (Ph), 127.46 (Ph), 127.4 (Ph),
126.9 (Ph), 126.1 (Ph), 114.3 (CH2@CH), 102.9 (C1), 101.7, 101.2,
100.9 (3C, PhCH, C10), 99.41 (C100), 79.9, 78.8, 78.0, 77.8, 77.1,
76.5, 75.5 (C20, C200, C3, C30, C300, C4, C40), 75.0 (PhCH2), 74.0
(PhCH2), 73.8 (C400), 72.7 (PhCH2), 70.7 (CH@CH2(CH2)5CH2O),
69.0, 68.5 (C6, C60), 66.8, 66.74, 66.70, 66.6 (C2, C5, C50, C500),
33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O), 28.83
(CH@CH2(CH2)5CH2O), 28.80 (CH@CH2(CH2)5CH2O), 25.8 (CH@CH2
(CH2)5CH2O), 17.04 (C600). ESIMS: m/z Calcd [C61H71N3O14]Na+:
1092.4828. Found: 1092.4823.
(500 MHz, D2O): dH 5.19 (1H, d, J1 ,2 = 4.0 Hz, H100), 4.65 (1H, d,
00 00
J1,2 = 7.7 Hz, H1), 4.41 (1H, d, J1 ,2 = 8.8 Hz, H10), 4.32 (1H, q,
0
0
J5 ,6 = 6.6 Hz, H500), 4.01–3.45 (17H, m, H2, H20, H200, H3, H30,
00 00
H300, H4, H40, H400, H5, H50, H6, H60, CH3(CH2)6CH2O), 2.06 (3H, s,
CH3C@O), 1.60–1.48 (2H, m, CH3(CH2)6CH2O), 1.37–1.26 (10H,
m, CH3(CH2)6CH2O), 1.24 (3H, d, 3H, J5 ,6 = 6.6 Hz, H600), 0.87
00 00
(3H, t, J = 7.0 Hz, CH3(CH2)6CH2O). 13C NMR (125 MHz, D2O): dC
175.0 (C@O), 103.4, 101.7, 101.0 (C1, C10, C100), 78.9, 78.2, 76.9,
76.6, 75.0, 73.3, 70.9, 70.7, 70.3, 69.6, 68.0 (C20, C200, C3, C30,
C300, C4, C40, C400, C5, C50, C500), 72.1 (CH3(CH2)6CH2O), 62.7, 62.3
(C6, C60), 56.4 (C2), 32.7 (CH3(CH2)6CH2O), 30.1 (CH3(CH2)6CH2O),
30.0
(CH3(CH2)6CH2O),
29.9
(CH3(CH2)6CH2O),
26.6
(CH3(CH2)6CH2O), 23.5 (CH3(CH2)6CH2O), 16.8 (C600), 14.9
(CH3(CH2)6CH2O). ESIMS: m/z Calcd [C28H51NO15]Na+: 664.3151.
Found: 664.3160.
3.12. 7-Octen-1-yl 2-acetamido-3-O-(4,6-O-benzylidene-2-O-
(2,3,4-tri-O-benzyl-a-L-fucopyranosyl)-b-D-galactopyranosyl)-2-
deoxy-b- -glucopyranoside (22)
D
3.14. 7-Octen-1-yl 2-azido-3-O-(3-O-(2-azido-2-deoxy-3,4,6-
tetra-O-acetyl-
(2,3,4-tri-O-benzyl-
deoxy-b- -glucopyranoside (25)
a
-
D
-galactopyranosyl)-4,6-O-benzylidene-2-O-
A solution of azide 21 (250 mg, 0.234 mmol) in dry pyridine
(2 mL) was treated with CH3COSH (4 mL), and the solution was
stirred (rt, 8d). The mixture was filtered, concentrated and sub-
jected to flash chromatography (EtOAc–CH2Cl2 1:1) to afford firstly
unreacted azide 21 (55 mg, 22%). Further elution yielded the de-
sired compound 22 contaminated with some acetylated alcohol.
The resulting oil was taken up in MeOH (15 mL) and treated with
a catalytic amount of NaOMe in MeOH, and the solution was stir-
red (rt, 12 h). The solution was neutralized with Amberlite IR 120
(H+) and filtered, and the residue was subjected to flash chroma-
tography (EtOAc–CH2Cl2 1:1) to afford 22 (155 mg, 62%) as a col-
a-
L
-fucopyranosyl)-b- -galactopyranosyl)-2-
D
D
A solution of acceptor 21 (621 mg, 0.58 mmol) and the trichlo-
roacetimidate (823 mg, 1.74 mmol) in dry Et2O (15 mL) was trea-
ted with 4 Å molecular sieves (rt, 1 h). The mixture was cooled
(ꢀ20 °C) and treated with TMSOTf (10
lowed to warm (0 °C). The mixture was treated with Et3N
(200 L), filtered, concentrated and subjected to flash chromatog-
raphy (EtOAc–CH2Cl2 3:97) to afford tetrasaccharide 25 as a col-
lL, 0.058 mmol) and al-
l
ourless oil (737 mg, 92%).
[a] +8.9 (c 0.1, CH2Cl2); Rf 0.25
(EtOAc–hexanes 2:3); 1H NMR (500 MHz): dH 7.55–7.20 (25H,
ourless oil. [
a
] ꢀ22.2 (c 0.1, CH2Cl2); Rf 0.81 (CH2Cl2–MeOH, 9:1);
1H NMR (500 MHz): dH 7.61–7.54 (4H, m, Ph), 7.39–7.15 (19H, m,
Ph), 7.07–7.01 (2H, m, Ph), 5.85–5.76 (1H, m, CH2@CH), 5.58 (1H,
00 00
m, Ph), 5.87–5.77 (1H, m, CH2@CH), 5.53 (1H, d, J1 ,2 = 2.5 Hz,
H100), 5.51 (1H, s, PhCH), 5.49 (1H, s, PhCH), 5.28 (1H, d,
s, PhCH), 5.56 (1H, s, PhCH), 5.11 (1H, d, J1 ,2 = 3.0 Hz, H100),
00 00
= 3.2 Hz, H1000), 5.20 (1H, dd, J2
= 11.0 Hz, J3
= 2.9 Hz,
000,2000
000,3000
000,4000
J1
H3000), 5.18–5.12 (2H, m, PhCH2, H4000), 5.04–4.93 (3H, m, PhCH2,
5.03–4.93 (3H, m, PhCH2, CH2@CH), 4.90 (1H, d, J1,2 = 8.2 Hz, H1),
4.79–4.67 (4H, m, PhCH2), 4.63 (1H, A of AB, J = 11.5 Hz, PhCH2),
CH2@CH), 4.90 (1H, A of AB, J = 11.9 Hz, PhCH2), 4.75 (2H, s,
4.39 (1H, d, J1 ,2 = 7.6 Hz, H10), 4.36 (1H, dd, J6,6 = 10.4 Hz,
J5,6 = 4.9 Hz, H6), 4.28–4.20 (2H, m, H3, H40), 4.15–4.04 (4H, m,
H200, H300, H500, H60), 3.97–3.88 (3H, m, H20, H60, OH), 3.87–3.78
(2H, m, H6, CH@CH2(CH2)5CH2O), 3.70 (1H, dd, J3,4 = J4,5 = 9.2 Hz,
H4), 3.66–3.62 (2H, m, H30, H400), 3.41–3.53 (3H, m, H2, H5,
CH@CH2(CH2)5CH2O), 3.16–3.14 (m, H50), 1.85 (3H, s, CH3C@O),
2.07–2.00 (2H, m, CH@CH2(CH2)5CH2O), 1.59–1.48 (2H, m,
CH@CH2(CH2)5CH2O), 1.40–1.25 (6H, m, CH@CH2(CH2)5CH2O),
0
0
PhCH2), 4.67 (1H, d, J1 ,2 = 7.9 Hz, H10), 4.63 (1H,
A
of AB,
0
0
J = 11.9 Hz, PhCH2), 4.52 (1H, q, J5 ,6 = 6.3 Hz, H500), 4.46 (1H, d,
J1,2 = 8.0 Hz, H1), 4.33 (1H, dd, J6,6 = 10.5 Hz, J5,6 = 4.7 Hz, H6),
4.28–4.25 (1H, m, H40), 4.22–4.12 (5H, m, H20, H200, H300, H5000,
00 00
H60), 3.88 (1H, d, J6 ,6 = 12.4 Hz, H60), 3.84–3.74 (5H, m, H30, H4,
H400, H6, H6000), 3.70 (1H, dd, J2,3 = J3,4 = 9.2 Hz, H3), 3.99–
3.92 (1H, m, CH@CH2(CH2)5CH2O), 3.65–3.60 (1H, m,
0
0
000,3000
000,2000
= 11.0 Hz, J1
CH@CH2(CH2)5CH2O), 3.55 (1H, dd, J2
= 3.2 Hz,
1.22 (d, 3H, J5 ,6 = 6.2 Hz, H600). 13C NMR (125 MHz): dC 170.6
00 00
H2000), 3.50–3.37 (2H, m, H2, H5), 3.22 (1H, dd, J6
= 11.5 Hz,
000,6000
J5
= 3.5 Hz, H6000), 3.10–3.07 (1H, m, H50), 2.10–2.06 (2H, m,
(C@O). 139.0 (CH2@CH), 138.5 (Ph), 138.4 (Ph), 138.1 (Ph), 137.5
(Ph), 136.8 (Ph), 129.0 (Ph), 128.9 (Ph), 128.6 (Ph), 128.44 (Ph),
128.41 (Ph), 128.3 (Ph), 128.2 (Ph), 128.15 (Ph), 128.07 (Ph),
128.1 (Ph), 127.7 (Ph), 127.6 (Ph), 127.4 (Ph), 126.7 (Ph), 126.1
(Ph), 114.2 (CH2@CH), 102.82 (C10), 101.5, 101.3, 101.0 (3C, C1,
PhCH), 99.0 (C100), 80.2, 79..0, 78.9, 77.5, 77.3, 77.2, 74.5, 72.4
(C20, C200, C3, C30, C300, C4, C40, C400), 74.8, 73.1 (3C, PhCH2), 69.9,
69.1, 68.8 (C6, C60, CH@CH2(CH2)5CH2O), 68.6, 66.3 (3C, C5, C50,
C500), 57.55 (C2), 33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2
(CH2)5CH2O), 28.9 (CH@CH2(CH2)5CH2O), 28.8 (CH@CH2(CH2)5
CH2O), 25.7 (CH@CH2(CH2)5CH2O), 23.1 (CH3C@O), 17.0 (C600).
ESIMS: m/z Calcd [C63H75NO15]Na+: 1108.5029. Found: 1108.5029.
000,6000
CH@CH2(CH2)5CH2O), 2.09 (3H, s, CH3C@O), 2.09 (3H, s, CH3C@O),
1.94 (3H, s, CH3C@O), 1.77–1.55 (2H, m, CH@CH2(CH2)5CH2O),
1.47–1.35 (6H, m, CH@CH2(CH2)5CH2O), 1.22 (3H, d, J5 ,6 = 6.3 Hz,
00 00
H600). 13C NMR (125 MHz): dC 170.3 (C@O), 169.7 (C@O), 169.4
(C@O), 139.4 (Ph), 139.0 (Ph), 138.81 (Ph), 138.79 (CH2@CH),
137.6 (Ph), 137.0 (Ph), 129.0 (Ph), 128.7 (Ph), 128.3 (Ph), 128.24
(Ph), 128.16 (Ph), 128.1 (Ph), 128.0 (Ph), 127.45 (Ph), 127.42
(Ph), 127.37 (Ph), 127.3 (Ph), 127.2 (Ph), 126.2 (Ph), 126.1 (Ph),
114.3 (CH2@CH), 102.9 (C1), 101.4, 101.2, 100.7 (3C, C10, PhCH),
97.9 (C100), 94.1 (C1000), 80.7 (C20), 79.7 (C3), 74.9 (PhCH2), 74.0
(PhCH2), 72.5 (PhCH2), 77.9, 77.8, 77.3, 76.0, 72.0 (C200, C30, C300,
C4, C400), 72.0 (C40), 70.8 (CH@CH2(CH2)5CH2O), 69.1, 68.6
(C6, C60), 68.8, 68.0 (C3000,C4000), 67.6 (C5000), 66.7, 66.4, 66.11,
66.09 (C2, C5, C50, C500), 62.7 (C6000), 57.9 (C2000), 33.7 (CH@CH2
(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O), 28.82 (CH@CH2(CH2)5
CH2O), 28.80 (CH@CH2(CH2)5CH2O), 25.8 (CH@CH2(CH2)5CH2O),
20.7 (CH3C@O), 20.62 (CH3C@O), 20.58 (CH3C@O), 16.9 (C600).
ESIMS: m/z Calcd [C71H84N5O19]Na+: 1405.5738. Found:
1405.5740.
3.13. 7-Octyl 2-acetamido-2-deoxy-3-O-(2-O-(
a-L-
fucopyranosyl)-b- -galactopyranosyl)-b- -glucopyranoside (23)
D
D
Octenyl glycoside 6 (82 mg, 0.128 mmol) was then taken up in
THF–H2O (1:1, 10 mL), treated with Pd–C (10%, 15 mg) and stirred
under a H2 atmosphere (48 h). The mixture was then filtered and
concentrated to afford the fully deprotected trisaccharide 23 as a

2318
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
3.15. 7-Octen-1-yl 2-acetamido-3-O-(3-O-(2-acetamido-2-
deoxy-3,4,6-tetra-O-acetyl- -galactopyranosyl)-4,6-O-
benzylidene-2-O-(2,3,4-tri-O-benzyl- -fucopyranosyl)-b-
galactopyranosyl)-2-deoxy-b- -glucopyranoside (26)
H50), 2.06–1.99 (2H, m, CH@CH2(CH2)5CH2O), 1.85 (3H, s, CH3C@O),
1.60–1.50 (2H, m, CH@CH2(CH2)5CH2O), 1.48 (3H, s, CH3C@O),
a
-D
00 00
a
-L
D-
1.41–1.28 (6H, m, CH@CH2(CH2)5CH2O), 1.28 (1H, d, J5 ,6 = 6.4 Hz,
D
H600). 13C NMR (125 MHz): dC 172.5 (C@O), 171.6 (C@O), 139.2
(Ph), 138.8 (Ph), 138.7 (Ph), 138.6 (CH2@CH), 138.3 (Ph), 137.6
(Ph), 128.6 (Ph), 128.3 (Ph), 128.0 (Ph), 127.9 (Ph), 127.84 (2C,
Ph), 128.78 (Ph), 127.7 (Ph), 127.2 (2C, Ph), 127.0 (Ph), 126.8 (Ph),
126.4 (Ph), 126.0 (2C, Ph), 113.4 (CH2@CH), 102.5, 101.3, 101.1,
100.9 (4C, C1, C10, PhCH), 98.0 (C100), 91.8 (C1000), 80.1 (C4), 74.9
(PhCH2), 72.7 (PhCH2), 71.9 (PhCH2), 79.7, 77.7, 76.28, 76.27, 75.3,
73.5, 71.4, 70.5 (C20, C200, C3, C300, C3000, C40, C400, C4000), 69.4 (C30),
69.6, 68.8, 68.3 (C6, C60, CH@CH2(CH2)5CH2O), 68.2, 66.8, 66.4,
66.0 (C5, C50, C500, C5000), 62.0 (C6000), 55.4 (C2), 49.7 (C2000), 33.4
(CH@CH2(CH2)5CH2O), 29.2 (CH@CH2(CH2)5CH2O), 28.7 (CH@CH2
(CH2)5CH2O), 28.5 (CH@CH2(CH2)5CH2O), 25.5 (CH@CH2(CH2)5
CH2O), 22.0 (CH3C@O), 21.0 (CH3C@O), 15.8 (C600). ESIMS: m/z Calcd
[C71H88N2O20]Na+: 1311.5823. Found: 1311.5819.
A solution of tetrasaccharide 25 (355 mg, 0.257 mmol) in pyri-
dine (2 mL) was treated with AcSH (4 mL), and the solution was
stirred (14 d). The mixture was filtered, concentrated and sub-
jected to flash chromatography (EtOAc–CH2Cl2 1:1) to afford 26
as a colourless oil (270 mg, 74%). [
a
] ꢀ8.9 (c 0.2, CH2Cl2); Rf 0.76
(MeOH–CH2Cl2 1:9); 1H NMR (500 MHz): dH 7.55–7.23, 7.20–7.10
(25H, m, Ph). 6.78 (1H, d, J = 7.3 Hz, NH), 5.85–5.76 (1H, m,
CH2@CH), 5.50 (1H, s, PhCH), 5.35 (1H, s, PhCH), 5.43 (1H, d,
00 00
000,4000
J3
= 2.1 Hz, H4000), 5.27 (1H, d, J1 ,2 = 4.1 Hz, H100), 5.20 (1H, d,
J = 9.8 Hz, NH000), 5.05–4.92 (7H, m, H1000, H3000, PhCH2, CH2@CH),
4.72 (1H, of AB, J = 11.6 Hz, PhCH2), 4.67 (1H, of AB,
A
A
J = 12.0 Hz, PhCH2), 4.60 (1H, A of AB, J = 11.1 Hz, PhCH2), 4.54–
4.46 (3H, m, H1, H2000, H5000), 4.38 (1H, d, J1 ,2 = 8.2 Hz, H10), 4.32
0
0
00 00
(1H, dd, J6,6 = 10.7 Hz, J5,6 = 4.8 Hz, H6), 4.13 (1H, q, J5 ,6 = 6.3 Hz,
3.17. 7-Octen-1-yl 2-acetamido-3-O-(4,6-O-benzylidene-3-O-
H500), 4.11–3.92, 3.89–3.74 (14H, 2ꢁm, H2, H20, H200, H3, H300, H4,
(2,3,4,6-tetra-O-benzyl-
benzyl- -fucopyranosyl)-b-
glucopyranoside (30)
a
-
D
-galactopyranosyl)-2-O-(2,3,4-tri-O-
H40, H400, H6, H60, H6000, CH@CH2(CH2)5CH2O), 3.69 (1H, d,
a-
L
D-galactopyranosyl)-2-deoxy-b-
D-
J6 ,6 = 12.6 Hz, H60), 3.62 (1H, dd, J2 ,3 = 9.6 Hz, J3 ,4 = 3.8 Hz, H30),
3.50–3.41 (2H, m, H5, CH@CH2(CH2)5CH2O), 2.76 (s, H50), 2.12
(3H, s, CH3C@O), 2.07–2.00 (2H, m, CH@CH2(CH2)5CH2O), 1.94
(3H, s, CH3C@O), 1.85 (3H, s, CH3C@O), 1.71 (3H, s, CH3C@O),
1.60–1.51 (2H, m, CH@CH2(CH2)5CH2O), 1.40–1.28 (2H, m,
CH@CH2(CH2)5CH2O), 1.27 (3H, s, CH3C@O), 1.22 (3H, d,
0
0
0
0
0
0
A solution of acceptor 21 (400 mg, 0.37 mmol) and trichloroace-
timidate 2834 (770 mg, 1.12 mmol) in dry Et2O (15 mL) was treated
with 4 Å molecular sieves (rt, 1 h). The mixture was cooled (ꢀ20 °C)
and treated with TMSOTf (10 lL, 0.058 mmol) and allowed to warm
J5 ,6 = 6.3 Hz, H600). 13C NMR (125 MHz): dC 170.6 (C@O), 170.3
(0 °C). The mixture was treated with Et3N (200 lL), filtered, concen-
00 00
(C@O), 170.2 (C@O), 170.0 (C@O), 169.8 (C@O), 139.7 (Ph), 139.1
(CH2@CH), 138.7 (Ph), 138.6 (Ph), 137.7 (Ph), 137.3 (Ph), 129.5
(Ph), 129.4 (Ph), 128.4 (Ph), 128.3 (2C, Ph), 128.21(Ph), 128.19
(Ph), 128.0 (Ph), 127.9 (Ph), 127.7 (Ph), 127.6 (Ph), 127.3 (Ph),
127.1 (Ph), 126.6 (Ph), 126.2 (Ph), 114.2 (CH2@CH), 102.7, 102.5,
101.9, 101.5 (4C, C1, C10, PhCH), 98.2, 92.3 (C100, C1000), 80.9, 80.0,
77.9, 77.5, 77.4, 76.5 (C20, C200, C3, C300, C4, C400), 75.0 (PhCH2),
74.1 (PhCH2), 73.4 (C30), 73.0 (PhCH2), 70.2 (C40), 69.7, 69.1, 69.0
(C6, C60, CH@CH2(CH2)5CH2O), 68.7, 67.7, 67.3, 66.8, 66.6, 65.5
(C3000, C4000, C5, C50, C500, C5000), 62.4 (C6000), 55.2 (C2), 46.9 (C2000),
33.7 (CH@CH2(CH2)5CH2O), 29.4 (CH@CH2(CH2)5CH2O), 28.84
trated and subjected to flash chromatography (EtOAc–hexanes 1:1)
to afford tetrasaccharide 29 as a colourless oil (440 mg, 73%). A
solution of tetrasaccharide (350 mg, 0.220 mmol) in dry pyridine
(4 mL) was treated with AcSH (2 mL) and stirred (rt, 8 d). Concen-
tration followed by flash chromatography (EtOAc–CH2Cl2 3:7)
afforded firstly unreacted 29 (70 mg, 20%). Further elution afforded
the desired 30 as a colourless oil (230 mg, 65%). [
a
] ꢀ16.0 (c 0.6,
CH2Cl2); Rf 0.57 (EtOAc–hexanes 1:1); 1H NMR (500 MHz): dH
7.55–7.48 (4H, m, Ph), 7.38–7.12 (33H, m, Ph), 7.10–6.97 (8H, m,
Ph), 6.79 (1H, d, J = 7.4 Hz, NH), 5.85–5.75 (1H, m, CH2@CH), 5.52
= 4.0 Hz, H1000),
000,2000
(1H, s, PhCH), 5.47 (1H, s, PhCH), 5.31 (1H, d, J1
5.23 (1H, d, J1 ,2 = 3.5 Hz, H100), 5.02–4.92 (2H, m, CH2@CH), 4.88
(1H, A of AB, J = 11.8 Hz, PhCH2), 4.83 (1H, A of AB, J = 11.3 Hz,
PhCH2), 4.68 (1H, A of AB, J = 12.0 Hz, PhCH2), 4.61–4.51 (4H, m,
H1, PhCH2), 4.49–4.29 (9H, m, H10, H6, PhCH2), 4.23 (1H, A of AB,
00 00
(CH@CH2(CH2)5CH2O),
28.81
(CH@CH2(CH2)5CH2O),
25.6
(CH@CH2(CH2)5CH2O), 22.9 (CH3C@O), 22.5 (CH3C@O), 20.69
(CH3C@O), 20.66 (CH3C@O), 20.6 (CH3C@O), 17.3 (C600). ESIMS:
m/z Calcd [C77H94N2O23]Na+: 1437.6140. Found: 1437.6138.
J = 11.8 Hz, PhCH2), 4.19 (1H, d, J3 ,4 = 3.3 Hz, H40), 4.15 (1H, dd,
0
0
J2 ,3 = 10.0 Hz, J3 ,4 = 2.6 Hz, H300), 4.10–3.95 (5H, m, H20, H200, H3,
H500, H60), 3.92 (1H, dd, J2 = 4.0 Hz, H2000), 3.88–
= 10.0 Hz, J1
00 00
00 00
3.16. 7-Octen-1-yl 2-acetamido-3-O-(3-O-(2-acetamido-2-
000,3000
000,2000
deoxy-
O-benzyl-
-glucopyranoside (27)
a
-
D
-galactopyranosyl)-4,6-O-benzylidene-2-O-(2,3,4-tri-
a
-
L
-fucopyranosyl)-b- -galactopyranosyl)-2-deoxy-b-
D
3.70 (9H, m, H2, H30, H3000, H400, H4000, H6, H60, H6000, CH@CH2
(CH2)5CH2O), 3.56–3.40 (4H, m, H4, H5, H6000, CH@CH2(CH2)5CH2O),
3.30 (1H, d, J = 1.8 Hz, H5000), 2.80 (1H, s, H50), 2.08–2.00 (2H, m,
CH@CH2(CH2)5CH2O), 1.72 (3H, s, CH3C@O), 1.61–1.49 (2H, m,
CH@CH2(CH2)5CH2O), 1.41–1.23 (6H, m, CH@CH2(CH2)5CH2O),
D
A solution of triacetate 26 (225 mg, 0.160 mmol) in CH3OH was
treated with a catalytic amount of NaOCH3 in CH3OH, and the solu-
tion was stirred (2 h). The solution was neutralized with Amberlite
IR 120 (H+) and filtered, and the residue was subjected to flash chro-
matography (EtOAc–CH2Cl2 1:1) to afford triol 27 as a colourless oil
1.16 (3H, d, J5 ,6 = 6.4 Hz, H600). 13C NMR (125 MHz): dC 170.4
00 00
(C@O), 139.1 (CH2@CH), 139.0 (Ph), 138.8 (Ph), 138.7 (Ph), 138.64
(Ph), 138.58 (Ph), 138.3 (Ph), 128.2 (Ph), 137.6 (Ph), 137.3 (Ph),
129.3 (Ph), 128.9 (Ph), 128.5 (Ph), 128.44 (Ph), 128.43 (2C, Ph),
128.40 (2C, Ph), 128.34 (Ph), 128.27 (Ph), 128.22 (Ph), 128.17
(Ph), 128.12 (Ph), 128.10 (Ph), 128.04 (Ph), 127.98 (Ph), 127.94
(Ph), 127.89 (Ph), 127.87 (Ph), 127.82 (Ph), 127.78 (2C, Ph),
127.67 (2C, Ph), 127.65 (Ph), 127.61 (Ph), 127.5 (Ph), 127.42 (Ph),
127.38 (Ph), 127.1 (Ph), 127.05 (2C, Ph), 127.02 (2C, Ph), 126.95
(Ph), 126.4 (Ph), 126.2 (Ph), 114.2 (CH2@CH), 102.9, 102.4, 101.9,
101.0 (4C, C1, C10, PhCH), 97.6 (C1000), 92.4 (C100), 80.9, 79.5, 78.3,
77.8, 77.3, 76.5, 71.0, 69.5 (12C, C20, C200, C2000, C3, C30, C300, C3000,
C4, C40, C400, C4000, C5), 74.8 (PhCH2), 74.4 (PhCH2), 73.6 (2C, PhCH2),
72.82 (PhCH2), 72.78 (PhCH2), 71.6 (PhCH2), 69.9, 69.6, 69. 2, 69.1
(C6, C60, C6000, CH@CH2(CH2)5CH2O), 67.4, 66.6, 65.8 (C50, C500, C5000),
(192 mg, 94%). [
a
] ꢀ12.9 (c 0.3, CH3OH); Rf 0.55 (CH3OH–CH2Cl2
1:9); 1H NMR (500 MHz, CD3OD): dH 7.57–7.18 (25H, m, Ph),
5.85–5.73 (1H, m, CH2@CH), 5.60 (1H, s, PhCH), 5.52 (1H, s, PhCH),
5.29 (1H, d, J1 ,2 = 3.8 Hz, H100), 5.05 (1H, d, J1
= 3.3 Hz, H1000),
00 00
000,2000
5.00–4.88 (4H, m, PhCH2, CH2@CH), 4.77 (1H, A of AB, J = 12.5 Hz,
PhCH2), 4.69 (1H, A of AB, J = 10.5 Hz, PhCH2), 4.67 (1H, d,
J1,2 = 6.7 Hz, H1), 4.83, 4.60 (2H, AB, J = 10.8 Hz, PhCH2), 4.49–4.35
(3H, m, H10, H500, H6), 4.33–4.06, 4.00–3.90, 3.87–3.79 (16H,
3 ꢁ m, H2, H20, H200, H2000, H3, H300, H3000, H40, H400, H4000, H5000, H6,
H60, CH@CH2(CH2)5CH2O), 3.73 (1H, dd, J3,4 = J4,5 = 9.2 Hz, H4),
3.58–3.53 (1H, m, H6000), 3.52–3.41 (3H, m, H3, H5,
´
CH@CH2(CH2)5CH2O), 3.28–3.22 (1H, m, H6000), 3.22–3.18 (1H, m,

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2319
55.5 (C2), 33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O),
28.84 (CH@CH2(CH2)5CH2O), 28.82 (CH@CH2(CH2)5CH2O), 25.6
(CH@CH2(CH2)5CH2O), 23.0 (CH3C@O), 17.1 (C600). ESIMS: m/z Calcd
[C97H109NO20]Na+: 1630.7435. Found: 1630.7455.
65.8 (C2), 33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O),
28.85 (CH@CH2(CH2)5CH2O), 28.81 (CH@CH2(CH2)5CH2O), 25.8
(CH@CH2(CH2)5CH2O). ESIMS: m/z Calcd [C28H37N3O5]Na+:
518.2625. Found: 518.2621.
3.18. 7-Octen-1-yl 2-azido-3-O-benzyl-4,6-O-benzylidene-2-
3.20. 7-Octen-1-yl 4-O-acetyl-2-azido-3,6-di-O-benzyl-2-deoxy-
deoxy-b-
D
-glucopyranoside (31)
b-D
-glucopyranoside (33)
A stirred solution of alcohol 13 (7.11 g, 17.6 mmol) in dry DMF
(50 mL) was cooled (ꢀ10 °C), the solution was treated with BnBr
(4.20 mL, 35.3 mmol) and NaH (60%, 960 mg, 24.0 mmol) and al-
lowed to warm (rt, 1 h). The mixture was then treated with CH3OH
(1 mL) and partially concentrated; the residuewas taken up in EtOAc
(250 mL) and washed with water and brine. The organic extract was
dried, concentrated and then subjected to flash chromatography
(EtOAc–hexanes 2:3) to afford benzyl ether 31 as a white crystalline
A solution of alcohol 32 (130 mg, 0.262 mmol) in pyridine
(5 mL) was treated with Ac2O (1 mL) and DMAP (5 mg), and the
solution was stirred (2 h). The excess Ac2O was quenched by the
addition of CH3OH (2 mL), and the solution was concentrated.
The residue was subjected to flash chromatography (EtOAc–hex-
anes 1:2) to afford 33 as a colourless oil (120 mg, 85%). [
a
] ꢀ18.3
(c 1.1, CH2Cl2); Rf 0.64 (EtOAc–hexanes 3:7). 1H NMR (400 MHz):
dH 7.39–7.26 (10H, m, Ph), 5.89–5.77 (1H, m, CH@CH2), 5.05–4.91
(3H, m, H4, CH@CH2), 4.83, 4.63 (2H, AB, J = 10.1 Hz, PhCH2), 4.53
(2H, s, PhCH2), 4.31 (1H, d, J1,2 = 7.8 Hz, H1), 3.95 (1H, ddd,
J = 9.4 Hz, 6.5 Hz, 6.5 Hz, CH@CH2(CH2)5CH2O), 3.60–3.37 (6H, m,
H2, H3, H5, H6, CH@CH2(CH2)5CH2O), 2.11–2.01 (2H, m,
CH@CH2(CH2)5CH2O), 1.86 (3H, s, CH3C@O), 1.72–1.63 (2H, m,
CH@CH2(CH2)5CH2O), 1.49–1.30 (6H, m, CH@CH2(CH2)5CH2O);
13C NMR (100.3 MHz): dC 167.12 (C@O), 136.5 (CH@CH2), 135.31
(Ph), 135.29 (Ph), 125.9 (Ph), 125.8 (Ph), 125.4 (Ph), 125.34 (Ph),
125.30 (Ph), 125.2 (Ph), 111.7 (CH@CH2), 99.6 (C1), 77.8 (C3),
72.3 (PhCH2), 71.1 (PhCH2), 71.0, 68.4 (C4, C5), 67.9, 67.1 (C6,
CH@CH2(CH2)5CH2O), 63.5 (C2), 31.2 (CH@CH2(CH2)5CH2O), 27.0
solid (8.50 g, 98%). Mp 53–55 °C; [
a
] ꢀ70.8 (c 1.5, CH2Cl2); Rf 0.81
(EtOAc–hexanes 3:7). 1H NMR (500 MHz): dH 7.52–7.47 (2H, m,
Ph), 7.43–7.28 (8H, m, Ph), 5.87–5.78 (1H, m, CH@CH2), 5.58 (1H, s,
PhCH), 5.05–4.99 (1H, m, CH@CH2), 4.97–4.94 (1H, m, CH@CH2),
4.93, 4.81 (2H, AB, J = 11.3 Hz, PhCH2), 5.01 (1H, d, J1,2 = 8.2 Hz,
H1), 4.35 (1H, dd, J6,6 = 10.7 Hz, J5,6 = 4.9 Hz, H6), 3.92 (1H, ddd,
J = 9.4 Hz, 6.9 Hz, 6.9 Hz, CH@CH2(CH2)5CH2O), 3.81 (1H, dd,
J6,6 = 10.7 Hz, J5,6 = 10.3 Hz, H6), 3.72 (1H, dd, J3,4 = 9.2 Hz,
J4,5 = 9.2 Hz, H4), 3.61–3.52 (2H, m, H3, CH@CH2(CH2)5CH2O), 3.46
(1H, dd, J2,3 = 9.5 Hz, J1,2 = 8.2 Hz, H2), 3.39 (1H, ddd, J5,6 = 10.3 Hz,
J4,5 = 9.2 Hz, J5,6 = 4.9 Hz, H5), 2.10–2.03 (2H, m, CH@CH2(CH2)5
CH2O), 1.72–1.61 (2H, m, CH@CH2(CH2)5CH2O), 1.46–1.31 (6H, m,
CH@CH2(CH2)5CH2O); 13C NMR (125.7 MHz): dC 139.0 (CH@CH2),
137.9 (Ph), 137.1 (Ph), 129.0 (Ph), 128.3 (Ph), 128.2 (Ph), 128.1
(Ph), 127.8 (Ph), 126.0 (Ph), 114.2 (CH@CH2), 102.6 (C1), 101.3
(PhCH), 81.5 (C4), 78.9 (C3), 74.8 (PhCH2), 70.6 (C6), 68.6
(CH@CH2(CH2)5CH2O), 66.3, 66.1 (C2, C5), 33.6 (CH@CH2(CH2)5
CH2O), 29.35 (CH@CH2(CH2)5CH2O), 28.8 (CH@CH2(CH2)5CH2O),
28.7 (CH@CH2(CH2)5CH2O), 25.7 (CH@CH2(CH2)5CH2O), ESIMS:
m/z Calcd [C28H35N3O5]Na+: 516.2469. Found: 516.2467.
(CH@CH2(CH2)5CH2O),
26.33
(CH@CH2(CH2)5CH2O),
26.30
(CH@CH2(CH2)5CH2O), 23.3 (CH@CH2(CH2)5CH2O), 18.3 (CH3C@O).
ESIMS: m/z Calcd [C30H39N3O6]Na+: 560.2731. Found: 560.2733.
3.21. 7-Octen-1-yl 2-azido-4-O-(4,6-O-benzylidene-b-D-
galactopyranosyl)-3,6-di-O-benzyl-2-deoxy-b-D-
glucopyranoside (35)
A solution of acceptor 32 (1.32 g, 2.67 mmol) in dry CH2Cl2
(20 mL) was stirred over 4 Å molecular sieves (1.5 g), and the mix-
ture was stirred (rt, 1 h). The solution was then cooled (ꢀ40 °C),
treated with TMSOTf (0.2 mL), followed by dropwise addition of
trichloroacetimidate 1532 (3.3 g, 6.7 mmol), and then the mixture
was allowed to warm (0 °C). The mixture was neutralized with
Et3N (2 mL), concentrated and subjected to flash chromatography
(EtOAc–hexanes 1:1) to afford a colourless oil that was immedi-
ately used in the next step. The colourless oil was taken up in
CH3OH (100 mL), treated with a solution of NaOCH3 in CH3OH
and stirred (rt, 3 h). The solution was neutralized with Amberlite
IR 120 (H+), filtered and subjected to flash chromatography
(EtOAc–hexanes 1:1) to afford diol 35 as a colourless oil (1.54 g,
3.19. 7-Octen-1-yl 2-azido-3,6-di-O-benzyl-2-deoxy-b-D-
glucopyranoside (32)
A solution of benzylidene acetal 31 (9.5 g, 19.3 mmol) in CH2Cl2
(200 mL) was treated with 4 Å molecular sieves (5 g), and the mix-
ture was stirred (rt, 1 h). The mixture was cooled (0 °C) and treated
with Et3SiH (15.4 mL, 96.7 mmol) and CF3COOH (7.45 mL,
96.7 mmol), and the mixture was allowed to slowly warm (rt,
2 h). The mixture was then neutralized with Et3N (10 mL), filtered
and then diluted with CH2Cl2 (500 mL). The organic extract was
washed with water and brine, dried and then filtered. Concentra-
tion followed by flash chromatography (EtOAc–hexanes 1:2) affor-
78%). [
a
] ꢀ37.5 (c 0.8, CH2Cl2); Rf 0.28 (EtOAc–hexanes 1:1); 1H
ded alcohol 32 as a colourless oil (7.95 g, 83%). [
a
] ꢀ27.3 (c 1.1,
NMR (500 MHz): dH 7.51–7.44 (4H, m, Ph), 7.39–7.21 (11H, m,
Ph), 5.86–5.78 (1H, m, CH2@CH), 5.47 (1H, s, PhCH), 5.06, 4.89
(2H, AB, J = 12.2 Hz, PhCH2), 5.04–4.99 (1H, m, CH2@CH), 4.97–
4.93 (1H, m, CH2@CH), 4.71, 4.57 (2H, AB, J = 12.2 Hz, PhCH2),
CH2Cl2); Rf 0.54 (EtOAc–hexanes 3:7). 1H NMR (500 MHz): dH
7.43–7.23 (10H, m, Ph), 5.87–5.77 (1H, m, CH@CH2), 5.03–4.98
(1H, m, CH@CH2), 4.96–4.93 (1H, m, CH@CH2), 4.92, 4.78 (2H, AB,
J = 11.0 Hz, PhCH2), 4.61, 4.58 (2H, AB, J = 12.6 Hz, PhCH2), 4.29
(1H, d, J1,2 = 8.5 Hz, H1), 3.91 (1H, ddd, J = 9.4 Hz, 6.5 Hz, 6.5 Hz,
CH@CH2(CH2)5CH2O), 3.77–3.70 (2H, m, H6), 3.63 (1H, ddd,
J3,4 = 9.3 Hz, J4,4 = 9.3 Hz, J = 2.3 Hz, H4), 3.54 (1H, ddd, J = 9.4 Hz,
6.8 Hz, 6.8 Hz, CH@CH2(CH2)5CH2O), 3.44–3.39 (1H, m, H5), 3.38
(1H, dd, J2,3 = 10.1 Hz, J1,2 = 8.5 Hz, H2), 3.25 (1H, dd,
J2,3 = 10.1 Hz, J3,4 = 9.3 Hz, H3), 2.67 (1H, d, J = 2.3 Hz, OH), 2.10–
2.02 (2H, m, CH@CH2(CH2)5CH2O), 1.69–1.61 (2H, m,
CH@CH2(CH2)5CH2O), 1.45–1.31 (6H, m, CH@CH2(CH2)5CH2O);
13C NMR (125.7 MHz): dC 139.1 (CH@CH2), 138.2 (Ph), 137.7 (Ph),
128.6 (Ph), 128.5 (Ph), 128.1 (Ph), 128.0 (Ph), 127.8 (Ph),
127.7(Ph), 114.2 (CH@CH2), 102.2 (C1), 82.5 (C3), 75.0 (PhCH2),
73.9 (C5), 73.7 (PhCH2), 72.1 (C4), 70.3 (C6, CH@CH2(CH2)5CH2O),
4.52 (1H, d, J1 ,2 = 7.8 Hz, H10), 4.41 (1H, d, J1,2 = 7.5 Hz, H1), 4.26
0
0
(1H, d, J6 ,6 = 12.5 Hz, H60), 4.06–4.00 (3H, m, H4, H40, H6), 3.92
(1H, J = 9.4 Hz, 6.5 Hz, 6.5 Hz, CH@CH2(CH2)5CH2O), 3.83–3.76
(2H, m, H6, H60), 3.67–3.61 (1H, m, H20), 3.58–3.51 (1H, m,
CH@CH2(CH2)5CH2O), 3.49–3.41 (5H, m, H2, H3, H30, H5, OH),
2.94 (1H, s, H50), 2.50 (1H, d, J = 8.8, OH), 2.11–2.03 (2H, m,
CH@CH2(CH2)5CH2O), 1.72–1.61 (2H, m, CH@CH2(CH2)5CH2O),
1.49–1.31 (6H, m, CH@CH2(CH2)5CH2O); 13C NMR (125.7 MHz):
dC 139.1 (CH2@CH), 138.5 (Ph), 137.72 (Ph), 137.67 (Ph), 129.1
(Ph), 128.4 (Ph), 128.3 (Ph), 128.22 (Ph), 128.17 (Ph), 127.9 (Ph),
127.8 (Ph), 127.7 (Ph), 126.4 (Ph), 114.2 (CH2@CH), 103.3 (PhCH),
102.3 (C1), 101.3 (C10), 82.0 (C3), 77.4 (C4), 75.3 (PhCH2), 75.1,
74.5, 72.8, 72.4 (C20, C30, C40, C5), 73.4 (PhCH2), 70.3
0
0

2320
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
(CH@CH2(CH2)5CH2O), 68.9 (C60), 68.2 (C6), 66.7, 66.4 (C2, C50),
33.7, (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O), 28.9
(CH@CH2(CH2)5CH2O), 28.8 (CH@CH2(CH2)5CH2O), 25.8 (CH@CH2
(CH2)5CH2O). ESIMS: m/z Calcd [C41H51N3O10]Na+: 768.3467.
Found: 768.3466.
CH@CH2(CH2)5CH2O), 1.49–1.32 (6H, m, CH@CH2(CH2)5CH2O),
1.16 (3H, d, J5 ,6 = 6.5 Hz, H600), 1.13 (9H, s, (CH3)3C); 13C NMR
00 00
(125.7 MHz): dC 178.0 (C@O), 139.1 (CH2@CH), 138.7 (Ph), 138.61
(Ph), 138.58 (Ph), 138.98 (Ph), 138.96 (Ph), 137.9 (Ph), 129.1 (Ph),
128.8 (Ph), 128.5 (Ph), 128.42 (Ph), 128.39 (Ph), 128.33 (Ph),
128.29 (Ph), 128.2 (Ph), 128.0 (Ph), 127.9 (Ph), 127.8 (Ph), 127.6
(Ph), 127.54 (Ph), 127.46 (Ph), 127.4 (Ph), 127.2 (Ph), 126.2 (Ph),
114.3 (CH2@CH), 102.1 (C1), 100.9, 100.7 (C10, PhCH), 97.2 (C100),
81.2 (C3), 79.2 (C300), 77.5 (C50), 76.6, 75.7, 75.6, 75.3, 72.8, 71.3
(C20, C200, C30, C4, C40, C400), 75.8 (PhCH2), 74.8 (PhCH2), 73.4 (PhCH2),
73.2 (PhCH2), 72.5 (PhCH2), 70.2 (CH@CH2(CH2)5CH2O), 68.7, 67.8
(C6, C60), 66.6, 66.3, 65.8 (C2, C5, C500), 38.9 ((CH3)3C), 33.7
(CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O), 28.9 (CH@CH2
(CH2)5CH2O), 28.8 (CH@CH2(CH2)5CH2O), 27.1 ((CH3)3C), 25.8
(CH@CH2(CH2)5CH2O), 16.7 (C600). ESIMS: m/z Calcd [C73H87N3O15]-
Na+: 1268.6029. Found: 1268.6027.
3.22. 7-Octen-1-yl 2-azido-4-O-(4,6-O-benzylidene-3-O-
pivaloyl-b-D-galactopyranosyl)-3,6-di-O-benzyl-2-deoxy-b-D-
glucopyranoside (36)
A solution of diol 35 (970 mg, 1.30 mmol) in dry pyridine
(10 mL) was treated with trimethylacetyl chloride (230 L,
l
1.8 mmol), and the solution was stirred (rt, 2 h). The solution
was concentrated, and the residue subjected to flash chromatogra-
phy (EtOAc–hexanes 1:3) to afford 36 as a colourless oil (949 mg,
88%). [
a
] ꢀ16.8 (c 0.4, CH2Cl2); Rf 0.59 (EtOAc–hexanes 3:7); 1H
NMR (400 MHz): dH 7.49–7.23 (15H, m, Ph), 5.90–5.77 (1H, m,
CH2@CH), 5.44 (1H, s, PhCH), 5.06–4.89 (4H, m, PhCH2, CH2@CH),
3.24. 7-Octen-1-yl 2-azido-4-O-(4,6-O-benzylidene-2-O-(2,3,4-
0
0
4.73, 4.58 (2H, AB, J = 12.2 Hz, PhCH2), 4.66 (1H, dd, J2 ,3 = 10.2 Hz,
tri-O-benzyl-
O-benzyl-2-deoxy-b-
a
-
L
-fucopyranosyl)-b-
D-galactopyranosyl)-3,6-di-
J3 ,4 = 3.7 Hz, H30), 4.62 (1H, d, J1 ,2 = 7.8 Hz, H10), 4.26 (1H,
D
-glucopyranoside (38)
0
0
0
0
J1,2 = 7.8 Hz, H1), 4.20 (1H, d, J3 ,4 = 3.7 Hz, H40), 4.08–3.86 (5H,
m, H20, H4, H6, H60, CH@CH2(CH2)5CH2O), 3.79 (1H, dd,
0
0
A solution of trisaccharide 37 (651 mg, 0.522 mmol) in CH3OH
(50 mL) was treated with a catalytic amount of LiOCH3 (100 mg),
and the solution heated at reflux (7 d). The solution was neutral-
ized with Amberlite IR 120 (H+), filtered and subjected to flash
chromatography (EtOAc–hexanes 1:2) to firstly afford unreacted
37 (60 mg, 9%). Further elution (EtOAc–hexanes 1:1) afforded alco-
J6 ,6 = 4.3 Hz, J5 ,6 = 2.0 Hz, H60), 3.76 (1H, dd, J6,6 = 5.3 Hz,
J5,6 = 2.0 Hz, H6), 3.55 (1H, ddd, J = 9.4 Hz, J = 6.8 Hz, J = 6.8 Hz,
CH@CH2(CH2)5CH2O), 3.48–3.40 (3H, m, H2, H3, H5), 3.28 (1H, br
s, OH), 2.90 (1H, s, H50), 2.12–2.02 (2H, m, CH@CH2(CH2)5CH2O),
1.75–1.64 (2H, m, CH@CH2(CH2)5CH2O), 1.51–1.33 (6H, m,
CH@CH2(CH2)5CH2O), 1.24 (9H, s, (CH3)3C) 13C NMR (100 MHz):
dC 178.6 (C@O), 139.3 (CH2@CH), 138.8 (Ph), 138.1 (Ph), 137.7
(Ph), 128.9 (Ph), 128.7 (Ph), 128.5 (Ph), 128.4 (Ph), 128.2 (Ph),
128.1 (Ph), 127.6 (Ph), 127.5 (Ph), 126.2 (Ph), 114.5 (CH2@CH),
103.9 (C10), 102.6 (C1), 100.6 (PhCH), 82.5 (C3), 77.7 (C4), 75.3
(PhCH2), 74.6, 73.7, 73.4, 69.7 (C20, C30, C40, C5), 73.9 (PhCH2),
70.5 (CH@CH2(CH2)5CH2O), 69.0, 68.7 (C6, C60), 66.8, 66.6 (C2,
C50), 39.2 (CH3)3C), 33.9 (CH@CH2(CH2)5CH2O), 29.7 (CH@CH2
(CH2)5CH2O), 29.1 (CH@CH2(CH2)5CH2O), 29.0 (CH@CH2(CH2)5
CH2O), 27.3 (CH3)3C), 26.0 (CH@CH2(CH2)5CH2O). ESIMS: m/z Calcd
[C46H59N3O11]Na+: 852.4042. Found: 852.4034.
0
0
0
0
hol 38 (503 mg, 83%) as a colourless oil. [
a
] ꢀ75.8 (c 0.3, CH2Cl2); Rf
0.19 (EtOAc–hexanes 3:7); 1H NMR (500 MHz): dH 7.57–7.53 (4H,
m, Ph), 7.41–7.06 (26H, m, Ph), 5.87–5.77 (1H, m, CH2@CH), 5.59
(1H, s, PhCH), 5.14, 4.61 (2H, AB, J = 10.4 Hz, PhCH2), 5.03–4.92
(2H, m, CH2@CH), 5.01, 4.66 (2H, AB, J = 11.7 Hz, PhCH2), 4.95
(1H, d, J1 ,2 = 3.7 Hz, H100), 4.82, 4.74 (2H, AB, J = 11.3 Hz, PhCH2),
00 00
4.80, 4.72 (2H, AB, J = 12.0 Hz, PhCH2), 4.69, 4.40 (2H, AB,
J = 11.3 Hz, PhCH2), 4.40 (1H, d, J1 ,2 = 8.1 Hz, H10), 4.32 (1H, d,
0
0
J6 ,6 = 12.0 Hz, H60), 4.17–4.11 (3H, m, H1, H40, OH), 4.08–4.03
0
0
(2H, m, H200, H4), 3.99 (1H, dd, J6 ,6 = 12.0 Hz, J5 ,6 = 1.3 Hz, H60),
3.92–3.84 (4H, m, H5, H500, H6, CH@CH2(CH2)5CH2O), 3.79 (1H,
0
0
0
0
dd, J2 ,3 = 9.8 Hz, J1 ,2 = 8.1 Hz, H20), 3.64–3.55 (3H, m, H30, H400,
H6), 3.49 (1H, ddd, J = 9.4 Hz, 6.8 Hz, 6.8 Hz, CH@CH2(CH2)5CH2O),
3.37 (1H, dd, J2,3 = 9.9 Hz, J1,2 = 8.2 Hz, H2), 3.27–3.19 (2H, m, H3,
H300), 3.12 (1H, s, H50), 2.10–2.01 (2H, m, CH@CH2(CH2)5CH2O),
1.70–1.60 (2H, m, CH@CH2(CH2)5CH2O), 1.45–1.29 (6H, m,
0
0
0
0
3.23. 7-Octen-1-yl 2-azido-4-O-(4,6-O-benzylidene-3-O-
pivaloyl-2-O-(2,3,4-tri-O-benzyl-a-L-fucopyranosyl)-b-D-
galactopyranosyl)-3,6-di-O-benzyl-2-deoxy-b-D-
glucopyranoside (37)
CH@CH2(CH2)5CH2O), 1.07 (3H, d, J5 ,6 = 6.5 Hz, H600); 13C NMR
00 00
A solution of alcohol 36 (920 mg, 1.11 mmol) in dry Et2O (10 mL)
was treated with 4 Å molecular sieves (500 mg), and the mixture
was stirred (rt, 1 h). The mixture was then cooled (ꢀ10 °C), treated
(125.7 MHz): dC 139.1 (CH2@CH), 138.6 (Ph), 138.5 (Ph), 138.13
(Ph), 138.11 (Ph), 138.08 (Ph), 137.3 (Ph), 129.2 (2C, Ph), 128.9
(Ph), 128.8 (2C, Ph), 128.4 (2C, Ph), 128.34 (Ph), 128.31 (Ph),
128.26 (Ph), 128.2 (Ph), 128.1 (2C, Ph), 127.74 (Ph), 127.65 (Ph),
127.5 (Ph), 126.5 (Ph), 114.2 (CH2@CH), 102.0 (C1), 101.4 (C10),
101.3 (PhCH), 99.6 (C100), 81.1 (C3), 79.0 (C5), 78.5 (C20), 77.5,
77.3, 75.9, 75.7, 75.0, 72.6 (C2 , C3 , C3 , C4, C4, C4 ), 75.8 (PhCH2),
74.8 (PhCH2), 74.3 (PhCH2), 73.4 (PhCH2), 73.0 (PhCH2), 70.1
(CH@CH2(CH2)5CH2O), 69.0, 67.9 (C6, C60), 67.5, 66.6, 65.8 (C2,
C50, C500), 33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O),
28.9 (CH@CH2(CH2)5CH2O), 28.8 (CH@CH2(CH2)5CH2O), 25.8
(CH@CH2(CH2)5CH2O), 16.8 (C600). ESIMS: m/z Calcd [C68H79N3O14]-
Na+: 1184.5454. Found: 1184.5441.
with TMSOTf (50 lL), followed by dropwise addition of trichloro-
acetimidate 1929 (1.87 g, 3.30 mmol) in dry Et2O (5 mL). The mix-
ture was treated with Et3N (0.5 mL), filtered and subjected to
flash chromatography (EtOAc–hexanes 1:3) to yield trisaccharide
00
0
00
00
´
37 as a colourless oil (1.30 g, 94%). [
a
] ꢀ66.7 (c 0.2, CH2Cl2); Rf 0.5
(EtOAc–hexanes 3:7); 1H NMR (500 MHz): dH 7.52–7.16 (30H, m,
Ph), 5.88–5.79 (1H, m, CH2@CH), 5.45 (1H, s, PhCH), 5.41 (1H, d,
J1 ,2 = 3.8 Hz, H100), 5.17 (1H, A of AB, J = 10.4 Hz, PhCH2), 5.05–
00 00
4.94 (3H, m, CH2@CH, PhCH2), 4.81 (1H, A of AB, J = 10.4 Hz, PhCH2),
4.80 (1H, dd, J2 ,3 = 10.2 Hz, J3 ,4 = 3.7 Hz, H30), 4.77–4.57 (6H, m,
0
0
0
0
PhCH2), 4.44 (1H, d, J1 ,2 = 7.9 Hz, H10), 4.38 (1H,
A of AB,
0
0
J = 12.0 Hz, PhCH2), 4.33 (1H, d, J3 ,4 = 4.0 Hz, H40), 4.28 (1H, dd,
3.25. 7-Octen-1-yl 2-acetamido-4-O-(4,6-O-benzylidene-2-O-
0
0
J6 ,6 = 12.4 Hz, J5 ,6 = 1.0 Hz, H60), 4.22–4.15 (3H, m, H1, H20, H500),
4.13–4.05 (2H, m, H200, H4), 3.96–3.90 (2H, m, H60,
CH@CH2(CH2)5CH2O), 3.83–3.76 (2H, m, H300, H6), 3.68 (1H, d,
(2,3,4-tri-O-benzyl-
a-
L-fucopyranosyl)-b-
D-galactopyranosyl)-
0
0
0
0
3,6-di-O-benzyl-2-deoxy-b-
D-glucopyranoside (39)
J3 ,4 = 2.1 Hz, H400), 3.60–3.51 (2H, m, H6, CH@CH2(CH2)5CH2O),
3.39 (1H, dd, J2,3 = 9.8 Hz, J1,2 = 8.2 Hz, H2), 3.21 (1H, dd,
J2,3 = 9.8 Hz, J2,3 = 9.0 Hz, H3), 3.13–3.09 (1H, m, H5), 3.02 (1H, s,
H50), 2.11–2.04 (2H, m, CH@CH2(CH2)5CH2O), 1.74–1.63 (2H, m,
A solution of trisaccharide 38 (500 mg, 0.431 mmol) in dry
pyridine (6 mL) was treated with AcSH (3 mL), and the solution
was stirred (rt, 8 d). The mixture was concentrated and then sub-
jected to flash chromatography (CH3OH–CH2Cl2 1:9) to afford 39
0
0

P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
2321
as a colourless oil (426 mg, 84%). [
a
] ꢀ54.5 (c 0.4, CH2Cl2); Rf 0.30
2834 (1.49 g, 3.15 mmol) in dry Et2O (15 mL) was then added drop-
wise, and the mixture was allowed to stand (20 min). The mixture
was neutralized with Et3N (0.5 mL), filtered, concentrated and sub-
jected to flash chromatography (EtOAc–hexanes 1:4) to afford the
nearly pure tetrasaccharide 41 (1.33 g, 75%) as a colourless oil. A
solution of tetrasaccharide 41 in dry pyridine (6 mL) was treated
with AcSH (3 mL), and the solution was stirred (rt, 2 d). The solu-
tion was then concentrated and subjected to flash chromatography
(CH3OH–CH2Cl2 1:9) to afford 42 (510 mg, 91%) as a colourless oil.
(EtOAc–hexanes 1:1); 1H NMR (500 MHz): dH 7.53–7.47 (4H, m,
Ph), 7.40–7.10 (26H, m, Ph), 5.84–5.74 (1H, m, CH2@CH), 5.71
00 00
(1H, d, J = 7.1 Hz, NH), 5.55 (1H, s, PhCH), 5.03 (1H, d, J1 ,2 = 3.5 Hz,
H100), 5.01–4.91 (4H, m, CH2@CH, PhCH2), 4.83, 4.71 (2H, AB,
J = 11.6 Hz, PhCH2), 4.78 (1H, A of AB, J = 12.4 Hz, PhCH2), 4.76
(1H, d, J1,2 = 8.2 Hz, H1), 4.73 (1H, A of AB, J = 12.0 Hz, PhCH2),
4.67–4.59 (3H, m, PhCH2), 4.43–4.39 (2H, m, H10, PhCH2), 4.26
(1H, d, J6 ,6 = 12.0 Hz, H60), 4.14 (1H, d, J3 ,4 = 3.5 Hz, H40), 4.07
0
0
0
0
(1H, dd, J2 ,3 = 10.1, J1 ,2 = 3.5 Hz, H200), 4.03 (1H, dd, J3,4 = J4,5
[a
] ꢀ7.8 (c 0.4, CH2Cl2); Rf 0.50 (CH3OH–CH2Cl2 1:20); 1H NMR
00 00
00 00
7.5 Hz, H4) 4.00–3.92 (3H, m, H3, H500, H60), 3.90 (1H, dd,
(500 MHz): dH 7.42–7.09 (48H, m, Ph), 7.03–6.98 (2H, m, Ph),
J2 ,3 = 10.1 Hz, J3 ,4 = 2.6 Hz, H300) 3.86–3.75 (3H, m, H20, H6,
CH@CH2(CH2)5CH2O), 3.72 (1H, dd, J6,6 = 10.5 Hz, J5,6 = 3.3 Hz, H6),
3.66–3.60 (2H, m, H30, H400) 3.56–3.46 (2H, m, H2, H5), 3.45–3.38
(1H, m, CH@CH2(CH2)5CH2O), 3.16 (1H, s, H50), 2.06–1.98 (2H, m,
CH@CH2(CH2)5CH2O), 1.84 (3H, s, CH3CO), 1.59–1.49 (2H, m,
CH@CH2(CH2)5CH2O), 1.41–1.23 (6H, m, CH@CH2(CH2)5CH2O),
5.87–5.76 (2H, m, CH2@CH, NH), 5.62 (1H, d, J1 ,2 = 3.6 Hz, H100),
00 00
00 00
00 00
5.38 (1H, s, PhCH), 5.36 (1H, d, J1
= 3.6 Hz, H1000), 5.02–4.97
000,2000
(2H, m, CH2@CH, PhCH2), 4.96–4.92 (2H, m, CH2@CH, PhCH2),
4.87–4.79 (4H, m, H1, PhCH2), 4.72 (1H, A of AB, J = 12.3 Hz,
PhCH2), 4.66–4.57 (6H, m, PhCH2), 4.52 (1H, A of AB, J = 12.0 Hz,
PhCH2), 4.47–4.30 (9H, m, H10, H500, H60, PhCH2), 4.18–4.12 (2H,
m, H20, H40), 4.08–3.91 (6H, m, H200, H2000, H3, H300, H4, H6), 3.87–
3.76 (5H, m, H30, H3000, H5000, H60, CH@CH2(CH2)5CH2O), 3.71 (1H,
s, H400), 3.59–3.54 (1H, m, H5), 3.53–3.40 (5H, m, H2, H4000, H6,
H6000, CH@CH2(CH2)5CH2O), 3.16–3.10 (1H, m, H6000), 2.92 (1H, s,
H50), 2.08–2.00 (2H, m, CH@CH2(CH2)5CH2O), 1.84 (3H, s, CH3CO),
1.63–1.53 (2H, m, CH@CH2(CH2)5CH2O), 1.41–1.23 (9H, m, H600,
CH@CH2(CH2)5CH2O). 13C NMR (125.7 MHz): dC 170.1 (C@O),
139.2 (Ph), 139.04 (CH2@CH), 138.99 (Ph), 138.96 (Ph), 138.76
(Ph), 138.72 (Ph), 138.66 (Ph), 138.5 (Ph), 138.4 (Ph), 138.3 (Ph),
137.8 (Ph), 128.8 (Ph), 128,6 (Ph), 128.4 (Ph), 128.34 (Ph), 128.32
(Ph), 128.25 (Ph), 128.21 (Ph), 128.18 (Ph), 128.08 (Ph), 128.06
(Ph), 127.98 (Ph), 127.65 (Ph), 127.59 (Ph), 127.5 (Ph), 127.45
(Ph), 127.40 (Ph), 127.37 (Ph), 127.24 (Ph), 127.15 (Ph), 127.07
(Ph), 126.3 (Ph), 114.2 (CH2@CH), 101.5, 101.1 (PhCH, C10), 99.8
(C1), 97.8 (C100), 92.8 (C1000), 80.0, 78.1, 78.0, 77.2, 76.32, 76.26,
76.1, 75.5, 75.3, 72.4, 72.2, 70.4 (13C, C20, C200, C2000, C3, C30, C300,
C3000, C4, C40, C400, C4000, C5, C5000), 75.0 (PhCH2), 74.5 (PhCH2), 74.2
(PhCH2), 73.5 (PhCH2), 73.4 (PhCH2), 73.2 (PhCH2), 72.8 (PhCH2),
72.3 (PhCH2), 71.3 (PhCH2), 69.8, 69.5, 69.1, 69.0 (C6, C60, C6000,
CH@CH2(CH2)5CH2O), 66.5 (C500), 66.2 (C50), 54.9 (C2), 33.7
(CH@CH2(CH2)5CH2O), 29.6 (CH@CH2(CH2)5CH2O), 28.91 (CH@CH2
(CH2)5CH2O), 28.89 (CH@CH2(CH2)5CH2O), 25.9 (CH@CH2(CH2)5
CH2O), 23.5 (CH3CO), 16.9 (C600). ESIMS: m/z Calcd [C104H117NO20]-
Na+: 1722.8061. Found: 1722.8063.
1.13 (3H, d, J5 ,6 = 6.4 Hz, H600); 13C NMR (125.7 MHz): dC 170.1
00 00
(C@O), 139.1 (CH2@CH), 138.8 (Ph), 138.62 (Ph), 138.58 (Ph),
138.3 (Ph), 138.0 (Ph), 137.5 (Ph), 129.0 (Ph), 128.7 (Ph), 128.6
(Ph), 128.4 (Ph), 128.3 (Ph), 128.24 (Ph), 128.22 (Ph), 128.18 (Ph),
128.1 (Ph), 128.0 (Ph), 127.7 (Ph), 127.59 (Ph), 127.57 (Ph), 127.5
(Ph), 127.4 (Ph), 126.5 (Ph), 114.2 (CH2@CH), 101.3 (PhCH), 101.0
(C10), 99.85, 99.78 (C1, C100), 79.1 (C300), 78.8 (C20), 77.6, 77.4 (C200,
C400), 76.1, 75.6, 72.9 (4C, C3, C30, C4, C500), 74.8 (PhCH2), 74.7
(C40), 74.1 (2C, PhCH2), 73.4 (PhCH2), 72.8 (PhCH2), 69.4, 69.0,
68.8 (C6, C60, CH@CH2(CH2)5CH2O), 67.6, 66.5 (C5, C50), 55.0 (C2),
33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O), 28.9 (2C,
CH@CH2(CH2)5CH2O), 25.8 (CH@CH2(CH2)5CH2O), 23.4 (CH3CO),
16.8 (C600). ESIMS: m/z Calcd [C70H83NO15]Na+: 1200.5655. Found:
1200.5661.
3.26. 7-Octyl 2-acetamido-4-O-(2-O-(
a-L-fucopyranosyl)-b-D-
galactopyranosyl)-2-deoxy-b- -glucopyranoside (40)
D
A solution of trisaccharide 7 (145 mg, 0.227 mmol) was taken
up in CH3OH, treated with Pd–C (10%, 20 mg) and subjected to a
H2 atmosphere (rt, 1 d). The mixture was then filtered and concen-
trated to afford octyl glycoside 40 as a colourless oil (140 mg, 96%).
[
a
] ꢀ106.1 (c 0.5, CH3OH); 1H NMR (500 MHz, CD3OD): dH 5.21 (1H,
d, J1 ,2 = 3.2 Hz, H100), 4.48 (1H, d, J1 ,2 = 7.1 Hz, H10), 4.37 (1H, d,
00 00
0
0
J1,2 = 8.4 Hz, H1), 4.17 (1H, q, J5 ,6 = 6.5 Hz, H500), 3.93–3.80, 3.79–
00 00
3.53, 3.34–3.30 (16H, 3 ꢁ m, H2, H20, H200, H3, H30, H300, H4, H40,
H400, H5, H50, H6, H60, CH3(CH2)6CH2O), 3.48–3.41 (1H, m,
CH3(CH2)6CH2O), 1.96 (3H, s, CH3CO), 1.57–1.48 (2H, m,
CH3(CH2)6CH2O)), 1.37–1.24 (10H, m, CH3(CH2)6CH2O), 1.20 (3H,
3.28. 7-Octen-1-yl 2-acetamido-4-O-(3-O-(2-acetamido-2-
deoxy-3,4,6-tri-O-acetyl-
a-
D-galactopyranosyl)-4,6-O-
benzylidene-2-O-(2,3,4-tri-O-benzyl-
a
-L
-fucopyranosyl)-b-D-
galactopyranosyl)-3,6-di-O-benzyl-2-deoxy-b-D-
glucopyranoside (44)
d, J5 ,6 = 6.5 Hz, H600), 0.89 (3H, t, J = 6.9 Hz, CH3(CH2)6CH2O). 13C
00 00
NMR (125.7 MHz, CD3OD): dC 173.4 (C@O), 102.9, 102.6 (C1, C10),
101.9 (C100), 79.1, 78.3, 77.1, 77.0, 75.3, 74.1, 73.6, 71.7, 70.8,
70.70 (C20, C200, C3, C30, C300, C4, C40, C400, C5, C500), 70.7
(CH3(CH2)6CH2O), 68.3 (C50), 62.6, 61.7 (C6, C60), 56.8 (C2), 33.0
A solution of acceptor 38 (1.17 g, 1.01 mmol) in dry Et2O
(45 mL) was treated with 4Å molecular sieves (1 g) and the mix-
ture was stirred (rt, 1 h). The mixture was then cooled (ꢀ10 °C),
(CH3(CH2)6CH2O),
30.7
(CH3(CH2)6CH2O),
30.5
(2C,
treated with TMSOTf (50 lL, 0.29 mmol); trichloroacetimidate
CH3(CH2)6CH2O), 27.2 (CH3(CH2)6CH2O), 23.7 (CH3(CH2)6CH2O),
23.0 (CH3CO), 16.8 (C600), 14.4 (CH3(CH2)6CH2O). ESIMS: m/z Calcd
[C28H51NO15]Na+: 664.3151. Found: 664.3156.
2435 (1.43 g, 0.965 mmol) in dry Et2O (15 mL) was then added
dropwise and the mixture was allowed to stand (20 min). The mix-
ture was neutralized with Et3N (0.5 mL), filtered, concentrated and
subjected to flash chromatography (EtOAc–hexanes 1:3) to afford
the nearly pure tetrasaccharide as a colourless oil (1.21 g, 94%).
The residue was taken up in pyridine (6 mL) and treated with AcSH
(3 mL) and the solution was stirred (7 d). The solution was concen-
trated and subjected to flash chromatography (CH2Cl2–CH3OH,
10:1) to afford 44 as a colourless oil (530 mg, 76%). Rf 0.65
(CH3OH–CH2Cl2 1:9); 1H NMR (500 MHz): dH 7.44–7.18 (30H, m,
3.27. 7-Octen-1-yl 2-acetamido-4-O-(4,6-O-benzylidene-3-O-
(2,3,4,6-tetra-O-benzyl-
benzyl- -fucopyranosyl)-b-
benzyl-2-deoxy-b- -glucopyranoside (42)
a
-D
-galactopyranosyl)-2-O-(2,3,4-tri-O-
a
-L
D
-galactopyranosyl)-3,6-di-O-
D
A solution of acceptor 38 (1.22 g, 1.05 mmol) in dry Et2O
(40 mL) was treated with 4 Å molecular sieves (1 g), and the mix-
ture was stirred (rt, 1 h). The mixture was then cooled (ꢀ10 °C)
00 00
Ph), 5.85–5.73 (2H, m, CH2@CH, NH), 5.45 (1H, d, J1 ,2 = 3.9 Hz,
H100), 5.41 (1H, s, PhCH), 5.38 (1H, J = 9.8 Hz, NH), 5.20 (1H, A of
AB, J = 12.3 Hz, PhCH2), 5.12 (1H, A of AB, J = 11.1 Hz, PhCH2),
and treated with TMSOTf (50 lL, 0.29 mmol); trichloroacetimidate

2322
P. J. Meloncelli, T. L. Lowary / Carbohydrate Research 345 (2010) 2305–2322
5.09 (1H, d, J1
= 3.7 Hz, H1000), 5.04–4.90 (7H, m, H1, H3000, H4000,
000,2000
References
CH2@CH, PhCH2), 4.71–4.55 (8H, m, H10, H2000, PhCH2), 4.45–4.38
(1H, m, H500), 4.32–4.06 (7H, m, H20, H200, H3, H4, H40, H5000, H60),
4.01–3.97 (1H, m, H60), 3.94–3.81 (4H, m, H30, H300, H6,
CH@CH2(CH2)5CH2O), 3.80–3.72 (2H, m, H400, H6), 3.67 (1H, dd,
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J6
= 11.5 Hz, J5
= 7.6 Hz, H6000), 3.52–3.41 (2H, m, H5,
000,6000
000,6000
CH@CH2(CH2)5CH2O), 3.33–3.26 (1H, m, H2), 3.20 (1H, s, H50),
= 3.7 Hz, H6000), 2.10–1.99 (5H,
000,6000
000,6000
3.09 (1H, dd, J6 = 11.5 Hz, J5
m, CH3CO, CH@CH2(CH2)5CH2O), 1.94 (3H, s, CH3CO), 1.87 (3H, s,
CH3CO), 1.42 (3H, s, CH3CO), 1.64–1.55 (2H, m, CH@CH2
(CH2)5CH2O), 1.41–1.24 (12H, m, CH3CO, H600, CH@CH2(CH2)5
CH2O). 13C NMR (125.7 MHz): dC 170.34 (2C, C@O), 170.32 (C@O),
170.1 (C@O), 170.0 (C@O), 139.4 (Ph), 139.0 (CH2@CH), 138.7
(Ph), 138.70 (Ph), 138.67 (Ph), 128.3 (Ph), 138.2 (Ph), 137.6 (Ph),
129.1 (Ph), 128.8 (Ph), 128.4 (Ph), 128.31 (Ph), 128.27 (Ph), 128.2
(Ph), 128.1 (Ph), 128.04 (Ph), 128.02 (Ph), 127.8 (Ph), 127.61 (Ph),
127.57 (Ph), 127.51 (Ph), 127.47 (Ph), 127.4 (Ph), 127.31 (Ph),
126.5 (Ph), 126.2 (Ph), 114.3 (CH2@CH), 101.2, 101.0 (PhCH, C10),
99.7 (C1), 98.6 (C100), 92.1 (C1000), 80.1, 77.7, 77.5, 77.0, 75.8, 75.3,
71.8, 70.4, 69.7, 69.0, 67.0, 66.2 (13C, C20, C200, C3, C30, C300, C3000,
C4, C40, C400, C4000, C5, C500, C5000), 74.9 (PhCH2), 73.7 (PhCH2), 73.4
(PhCH2), 73.3 (PhCH2), 72.1 (PhCH2), 68.7, 68.4, 67.4 (C6, C60,
CH@CH2(CH2)5CH2O), 67.7 (C50), 63.0 (C6000), 56.6 (C2), 46.5 (C2000),
33.7 (CH@CH2(CH2)5CH2O), 29.5 (CH@CH2(CH2)5CH2O), 28.9 (2C,
CH@CH2(CH2)5CH2O), 25.9 (CH@CH2(CH2)5CH2O), 23.5 (CH3CO),
22.6 (CH3CO), 20.69 (CH3CO), 20.66 (CH3CO), 20.65 (CH3CO), 16.8
(C600). ESIMS: m/z Calcd [C84H102NO23]Na+: 1529.6766. Found:
1529.6744.
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T. L.; Buriak, J. M., in preparation.
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Funding was provided by a Canadian Institutes of Health Re-
search (CIHR) team grant (RMF 92091), a Natural Sciences and
Engineering Research Council of Canada (NSERC) and Canadian
Institutes of Health Research (CIHR) CHRP grant (CHRPJ350946-
08), and the Alberta Ingenuity Centre for Carbohydrate Science.
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Supplementary data
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Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.carres.2010.08.012.