Synthesis of a heparan sulfate mimetic disaccharide with a conformationally locked residue from a common intermediate

Article abstract of DOI:10.1016/j.carres.2009.09.012

A simple mimetic of a heparan sulfate disaccharide sequence that binds to the growth factors FGF-1 and FGF-2 was synthesized by coupling a 2-azido-2-deoxy-d-glucopyranosyl trichloroacetimidate donor with a 1,6-anhydro-2-azido-2-deoxy-β-d-glucopyranose acceptor. Both the donor and acceptor were obtained from a common intermediate readily obtained from d-glucal. Molecular docking calculations showed that the predicted locations of the disaccharide sulfo groups in the binding site of FGF-1 and FGF-2 are similar to the positions observed for co-crystallized heparin-derived oligosaccharides obtained from published crystal structures.

Full text of DOI:10.1016/j.carres.2009.09.012

Carbohydrate Research 344 (2009) 2394–2398
Contents lists available at ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Note
Synthesis of a heparan sulfate mimetic disaccharide with a conformationally
locked residue from a common intermediate
à
,§
*
Jon K. Fairweather , Tomislav Karoli, Ligong Liu , Ian Bytheway, Vito Ferro
Drug Design Group, Progen Pharmaceuticals Limited, Brisbane, Qld 4066, Australia
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 6 July 2009
Received in revised form 11 September
2009
Accepted 14 September 2009
Available online 19 September 2009
A simple mimetic of a heparan sulfate disaccharide sequence that binds to the growth factors FGF-1 and
FGF-2 was synthesized by coupling a 2-azido-2-deoxy- -glucopyranosyl trichloroacetimidate donor with
a 1,6-anhydro-2-azido-2-deoxy-b- -glucopyranose acceptor. Both the donor and acceptor were obtained
from a common intermediate readily obtained from -glucal. Molecular docking calculations showed that
the predicted locations of the disaccharide sulfo groups in the binding site of FGF-1 and FGF-2 are similar
to the positions observed for co-crystallized heparin-derived oligosaccharides obtained from published
crystal structures.
D
D
D
Keywords:
Heparan sulfate mimetics
Disaccharides
Ó 2009 Elsevier Ltd. All rights reserved.
Fibroblast growth factors
The fibroblast growth factors FGF-1 and FGF-2 are proteins that
play important roles in tumor angiogenesis.1 They initiate this pro-
cess by binding with their receptors (FGFRs) and heparan sulfate
(HS) to form a ternary complex which leads to receptor dimeriza-
tion/activation and subsequent cell signaling.2 Inhibiting the for-
mation of the HS–FGF–FGFR complex by antagonizing HS–FGF
binding with HS mimetics is thus a viable strategy for antiangio-
genic therapies.3–5
two key sulfo groups [GlcN(2S) and IdoA(2S)], the compounds were
designed to mimic the conformational flexibility14,15 of the IdoA
residue. Docking calculations showed that the predicted locations
of disaccharide sulfo groups in the binding site of FGF-1 were con-
sistent with the positions observed for co-crystallized heparin-
derived oligosaccharides. Docking scores correlated with
experimental Kd values (22 lM to 1.4 mM) obtained from binding
assays.12 The docking score for a model HS disaccharide binding to
FGF-1 was similar.12
Various groups have reported the synthesis of HS and HS-like
oligosaccharides designed to interact with FGF-1 or FGF-2.6–8
These studies have provided valuable information about the struc-
tural requirements for oligosaccharide-FGF binding and activation,
however, the syntheses of such oligosaccharides are difficult and
laborious. This has lead to the pursuit of less synthetically chal-
lenging oligosaccharide mimetics as FGF antagonists.9–12 As part
of a program aimed at developing antiangiogenic compounds, we
recently described12 the preparation of simple disaccharides such
as 2 and 3 which mimic the HS disaccharide GlcN(2S, 6S)-IdoA(2S)
(1, Fig. 1), which has been postulated from X-ray crystallographic
analyses as a minimal heparin/HS consensus sequence for FGF
In crystal structures of heparin oligosaccharides bound to FGF,
IdoA is found in the 1C4 conformation when bound only to the pro-
tein16,17 or in a skew-boat (2SO) conformation when part of a ternary
complex.18,19 NMR studies also indicate that FGF-1 can bind both
conformations of IdoA in a bioactive hexasaccharide.20 These obser-
vations led us to consider the synthesis of simple disaccharides in
OSO3Na
OH
HO
O
O
O
R
binding.13 As well as maintaining the
a-(1?4) linkage between
HO
HO
NaO3SO
OSO3Na
OMe
O
HN
OH
O
the two monosaccharide units and the spatial orientation of the
O
O
NaO3S
O
NaO3SO
NaO3SO
* Corresponding author. Tel.: +617 3138 1108; fax: +617 3138 1804.
E-mail addresses: vito.ferro@qut.edu.au, v.ferro@griffith.edu.au (V. Ferro).
Present address: Chemical Analysis Pty Ltd, Bulleen, Vic. 3105, Australia.
1
2 R = CH2OSO3Na
3 R = H
à
Present address: Centre for Drug Design and Development, Institute for Molecular
Bioscience, University of Queensland, Brisbane, Qld 4072, Australia.
§
Present addresses: School of Physical and Chemical Sciences, Queensland
Figure 1. Structures of the GlcN(2S, 6S)-IdoA(2S) disaccharide sequence 1, which
represents a minimal consensus sequence for FGF–HS binding,13 and two conform-
ationally flexible mimetics 2 and 3.12
University of Technology, GPO Box 2434, Brisbane, Qld 4001, Australia; Institute for
Glycomics, Griffith University, Gold Coast Campus, Qld 4222, Australia.
0008-6215/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2009.09.012

J. K. Fairweather et al. / Carbohydrate Research 344 (2009) 2394–2398
2395
whichthe IdoAmimic is locked in the 1C4 conformation. A similarap-
proach has been successfully used to probe the active conformations
of the ATIII-binding heparin pentasaccharide.21,22
alyst and the resulting diamine 14 was sulfonated with SO3Átri-
methylamine complex and debenzoylated (1 M NaOH) to give
the benzyl ether 16 in moderate overall yield (28%, three steps).
Hydrogenolysis over Pd(OH)2 at 50 psi then furnished the target
disaccharide 17 in excellent yield (98%).
Molecular docking calculations were performed using the GLIDE
program28 to examine the binding modes of 16 and 17 with FGF-
1 and FGF-2. Compound 16 was examined in order to probe the ef-
fects of an extra hydrophobic group on FGF binding as it has been
shown that some heparin derivatives with lipophilic modifications
can bind to FGF-1 with similar or greater affinity than unmodified
heparin.29,30 The poses of 16 and 17 with the best GlideScores for
binding to FGF-1 and FGF-2 are shown in Figure 3. Also shown
are the van der Waals surfaces of the central sulfo groups of cocrys-
tallized, heparin-derived hexa- and tetrasaccharide ligands from
the crystal structures (pdb accession codes 2AXM16 for FGF-1 and
1BFB17 for FGF-2, respectively).
The preferred mode of binding of 16 and 17 to FGF-1 in Figure
3a shows congruence between the ligand sulfo groups with those
observed crystallographically. The FGF-2 binding mode of 16,
shown in Figure 3b, also shows the same congruence; however,
the preferred mode for 17, involves ionic hydrogen bonding inter-
actions with the positively charged residues LYS130 and LYS120 of
FGF-2. These residues are not involved in binding to the cocrystal-
lized heparin tetrasaccharide fragment, although their proximity to
the binding site region and their inherent flexibility suggests that
their involvement in ligand binding is reasonable. In the absence
It has been demonstrated that an O-sulfo group can substitute
for an N-sulfo group in a heparin oligosaccharide without loss of
binding affinity.23 The known 1,6-anhydro-2-azido-2-deoxy-b-
D
-
glucopyranose 8 was thus identified as a suitable IdoA mimic pre-
cursor because it is locked in the required 1C4 conformation, can be
selectively sulfonated at the C-2 position and is readily available in
four steps from D
-glucal (4) via intermediates 5–7 (Fig. 2).24 The
precursor to 8 in this sequence of transformations can, in fact, be
easily converted, via intermediates 9–11, into useful glycosyl do-
nors such as the imidate 12 for use in the synthesis of heparin/
HS oligosaccharides.24–26 It was therefore decided to prepare both
8 and 12 and use them to synthesize a disaccharide (17) with the
desired features.
Following literature precedent,27 TBDMSOTf was selected as the
promoter for the glycosylation of the alcohol 8 with the imidate 12.
The reaction proceeded well in dichloromethane at À20 °C; how-
ever, a chromatographically inseparable mixture of anomers re-
sulted (
tribenzoate via Zémplen deacetylation followed by benzoylation
with benzoyl chloride and pyridine, and the desired -linked disac-
a/b = 3.8:1). The product was thus converted into the
a
charide 13 was isolated by flash chromatography in good overall
yield (46%, three steps, Scheme 1), along with 12% of the b-linked
disaccharide 13b. The azide groups of 13 were then reduced via
transfer hydrogenation with ammonium formate over Pd(OH)2 cat-
OAc
O
OH
OR2
O
O
HO
HO
O
AcO
AcO
OR
N3
R3O
R1
4
5 R1 = I, R2 = R3 = H
10 R = Ac
11 R = OH
12 R = C(=NH)CCl3
6 R1 = N3, R2 = R3 = H
7 R1 = N3, R2 = R3 = Bn
8 R1 = N3, R2 = Bn, R3 = H
9 R1 = N3, R2 = R3 = A c
Figure 2. The structures of the glycosyl acceptor and glycosyl donor and their intermediates used in this study.
OBz
O
OBz
O
a, b
8 + 12
c
BzO
BzO
BzO
BzO
OBn
O
OBn
O
O
O
N3
H2N
O
O
d
N3
H2N
13
14
OH
OBz
O
HO
HO
e
O
BzO
BzO
OR
O
OBn
O
HN
SO3Na
O
O
HN
O
O
SO3Na
HN
SO3Na
HN
SO3Na
15
16 R = Bn
17 R = H
f
Scheme 1. Reagents and conditions: (a) TBDMSOTf, CH2Cl2, À20 °C; (b) (i) NaOMe, MeOH, (ii) BzCl, pyridine, 46%, three steps; (c) Pd(OH)2, NH4HCO2, EtOAc–MeOH, 58%; (d)
SO3ÁMe3N, DMF, 60 °C; (e) 1 M NaOH, 28%, three steps; (f) H2, Pd(OH)2, 98%.

2396
J. K. Fairweather et al. / Carbohydrate Research 344 (2009) 2394–2398
1.2. 3,4,6-Tri-O-acetyl-2-azido-2-deoxy-D-glucopyranosyl
trichloroacetimidate (12)
(a) A mixture of
D-glucal (1.95 g, 13.3 mmol), (n-Bu3Sn)2O
(5.44 mL, 10.6 mmol), and molecular sieves (4.0 g of 3 Å powder)
in MeCN (50 mL) was heated under reflux (3 h). The mixture was
cooled (rt) and I2 (4.06 g, 16.0 mmol) was introduced with continu-
ous stirring (1.5 h). The mixture was filtered and the filtrate was
washed with hexane (3 Â 100 mL) prior to evaporation of the sol-
vent affording, presumably, the iodide 5 as a pale yellow oil. This
was used directly in the next reaction without further purification
or characterization. (b) A mixture of the crude iodide and NaN3
(2.59 g, 39.9 mmol) in a mixture of DMF (45 mL) and H2O (5 mL)
was heated under reflux (o/n) and then evaporated and co-evapo-
rated (toluene). The residue was treated with pyridine (10 mL),
Ac2O (5 mL), and DMAP (50 mg) and the combined mixture was
stirred (rt, o/n). The mixture was then treated with ice-water
(10 mL) and stirring continued (3 h) before being subjected to
workup (EtOAc) and flash chromatography (10?40% EtOAc–hex-
ane) to yield the diacetate 9 as a colorless oil (1.90 g), used in the
next reaction without further purification or characterization. (c)
Concd H2SO4 (50 lL) was added to a cooled (0 °C) solution of crude
9 (2.33 g, 8.53 mmol) and Ac2O (5.00 mL, 49.0 mmol) in AcOH
(10 mL) and the combined mixture was stirred (0 °C?rt, o/n). So-
dium acetate (500 mg) was added portionwise until pH >5.0 and
then the mixture was treated with MeOH (3 mL). The mixture
was filtered and the solvent was evaporated and co-evaporated
(toluene) prior to workup (EtOAc) to yield a colorless oil (3.04 g).
This residue was co-evaporated (MeCN, 2 Â 100 mL) and used in
the next reaction without further purification or characterization.
(d) Hydrazine acetate (921 mg, 10.0 mmol) was added to a stirred
solution of the crude mixture from (c) (8.53 mmol, max.) in DMF
(20 mL) and the combined mixture was heated (55 °C, 15 min).
The mixture was poured onto saturated brine and extracted
(EtOAc). The organic layer was evaporated and subjected to rapid
silica filtration (10?40% EtOAc–hexane) to yield, presumably, the
hemiacetal 11 as a colorless oil (1.89 g, 76%, two steps). This residue
was co-evaporated (MeCN, 2 Â 100 mL) and used in the next reac-
tion without further purification or characterization. (e) K2CO3
(1.30 g, 9.5 mmol) was added to a solution of the hemiacetal 11
(1.85 g, 6.3 mmol) and trichloroacetonitrile (2.0 mL, 20 mmol) in
1,2-DCE (20 mL) and the combined mixture was stirred (0 °C?rt,
1 h). The mixture was filtered, the solvent was evaporated, and
the residue was subjected to flash chromatography (10–30%
Figure 3. The best Glide poses for 16 (gray C atoms) and 17 (green C atoms) docked
with FGF-1 (represented by a blue
a-carbon backbone) and FGF-2 (represented by
an orange -carbon backbone). Also shown are the van der Waals surfaces of the
a
sulfo groups of the heparin fragments cocrystallized with each growth factor (red O
atoms and yellow S atoms). (a) The best poses for 16 (Gscore = À9.2 kcal/mol) and
17 (Gscore = À7.5 kcal/mol) docked with FGF-1; (b) The best poses for 16
(Gscore = À7.2 kcal/mol) and 17 (Gscore = À6.1 kcal/mol) docked with FGF-2.
Hydrogen bonding interactions are depicted by the dotted yellow lines. (For
interpretation of the references to color in this figure legend, the reader is referred
to the web version of this article.)
of confirmatory experimental binding data these results are not
definitive. However, they do suggest that conformationally locked
residues and hydrophobic substituents29,30 should be further ex-
plored in HS mimetic design to target amino acid residues adjacent
to the HS-binding site of FGF-1 and FGF-2.
EtOAc–hexane) to yield an anomeric mixture (a/b = 9:1) of the imi-
In conclusion, a simple conformationally locked HS mimetic
disaccharide was efficiently synthesized via the coupling of a 2-azi-
date 12 as a colorless oil (2.19 g, 80%). This mixture was separable
by more careful chromatography and gave 1H NMR data in good
agreement with the literature.33,34
a
-Imidate: 1H NMR (400 MHz,
do-2-deoxy-
1,6-anhydro-2-azido-2-deoxy-b-
the donor and acceptor were obtained from a common intermedi-
ate readily obtained from -glucal. Molecular docking calculations
D-glucopyranosyl trichloroacetimidate donor with a
D-glucopyranose acceptor. Both
CDCl3) d 2.02, 2.03, 2.08 (3s, 3H each; 3 Â Ac), 3.74 (dd, 1H,
J1,2 = 3.6, J2,3 = 10.5 Hz; H-2); 4.06 (dd, 1H, J5,6a = 2.1, J6a,6b = 12.4 Hz;
H-6a), 4.18 (ddd, 1H, J4,5 = 10.3, J5,6b = 4.3 Hz; H-5), 4.21 (dd, 1H, H-
6b), 5.12 (dd, 1H, J3,4 = 9.4 Hz; H-4), 5.48 (dd, 1H, H-3), 6.46 (d, 1H,
H-1), 8.80 (s, 1H; NH). b-Imidate: partial 1H NMR (400 MHz, CDCl3)
d 5.68 (d, 1H, J1,2 = 8.4 Hz; H-1).
D
indicated that the predicted locations of the disaccharide sulfo
groups in the binding site of FGF-1 and FGF-2 are similar to the
positions observed for co-crystallized heparin-derived oligosaccha-
rides. These results may aid in the design of potential inhibitors of
FGF-mediated angiogenesis.
1.3. 1,6-Anhydro-2-azido-3-O-benzyl-2-deoxy-b-D-
glucopyranose (8)
1. Experimental
The alcohol 8 was prepared according to the literature proce-
dure24,35 by selective debenzylation of the dibenzyl ether 7 with
TiCl4. 1H NMR (400 MHz, CDCl3) d 1.96 (br s, 1H; OH), 3.48–3.49
(m, 1H; H-2), 3.56–3.58 (m, 1H; H-3), 3.64–3.66 (m, 1H; H-4),
3.75 (dd, 1H, J5,6a = 5.9, J6a,6b = 7.3 Hz; H-6a), 4.17 (dd, 1H,
J5,6b = 1.1, H-6b), 4.50–4.52 (m, 1H; H-5), 4.58, 4.61 (AB,
JA,B = 11.9 Hz; CH2Ph), 5.42–5.43 (m, 1H; H-1), 7.26–7.36 (m, 5H;
Ph).
1.1. General methods
General experimental details have been given previously.31
Capillary electrophoresis (CE) was performed in reverse polarity
mode with inverse UV detection, using 10 mM 5-sulfosalicylic
acid (pH 3) as the background electrolyte as described
previously.32

J. K. Fairweather et al. / Carbohydrate Research 344 (2009) 2394–2398
2397
1.4. 2-Azido-3,4,6-tri-O-benzoyl-2-deoxy-
(1?4)-1,6-anhydro-2-azido-3-O-benzyl-2-deoxy-b-
glucopyranose (13)
a
-
D
-glucopyranosyl-
Without further purification, to the diamine were added DMF
(5 mL), SO3ÁMe3N (41 mg, 295 mol), and NaHCO3 (40 mg,
475 mol). The mixture was heated (60 °C) for 1 h then cooled to
D
-
l
l
rt and the reaction was quenched with ice and Na2CO3 (satd aq).
This suspension was stored at À18 °C overnight and the sample
was filtered and evaporated. TLC indicated that partial hydrolysis
of the benzoate groups had occurred (presumably in the workup
(a) A solution of the imidate 12 (201 mg, 453
lmol) and alcohol
8 (84 mg, 302 mol) in 1,2-DCE (5 mL) was stirred in the presence
l
of activated mol. sieves (300 mg of 3 Å powder) under an atmo-
sphere of argon (rt, 30 min). The mixture was cooled (À20 °C) with
procedure). Water (1 mL) and NaOH (250 lL, 1 M) were added
continuous stirring (10 min) and TBDMSOTf (21
l
L, 91
lmol) was
and the solution was stirred overnight, then loaded directly onto
a Bio-Gel P-2 (5 Â 100 cm) column and was purified by size exclu-
sion chromatography (0.1 M ammonium bicarbonate, flow
rate = 2.8 mL/min). Fractions containing carbohydrate (determined
by spotting onto silica gel plates and visualization by charring)
were checked for purity by CE and those deemed to be free of salt
were pooled and lyophilized to give disulfate 16 (22 mg, 28%, three
steps) as an amorphous white solid. 1H NMR (400 MHz, D2O, sol-
vent suppressed) d 7.35–7.21 (m, 5H, Ph), 5.43 (br s, 1H, H-1I),
5.18 (d, 1H, J1,2 = 3.6 Hz, H-1II), 4.72–4.69 (m, 1H, H-5I), 4.54–4.52
(m, 2H, PhCH2), 4.05 (d, 1H, Jgem = 7.9 Hz, H-6aI), 3.85 (br s, 1H,
H-3I), 3.76–3.58 (m, 5H, H4I, H6bI, H5II, H6aII, 6bII), 3.51 (dd, 1H,
J2-3 = 10.4, J3-4 = 9.1, H-3II), 3.34 (dd, 1H, J3-4ꢀ4-5 = 9.2, H-4II), 3.23
(br s, 1H, H-2I), 3.12 (dd, 1H, H-2II); 13C NMR (100 MHz, CDCl3) d
133.3, 124.8, 124.6, 124.4, 96.9, 95.1, 72.7, 71.5, 70.8, 68.3, 68.2,
67.2, 66.0, 61.0, 56.6, 54.0, 49.8. The purity was 99% by CE
(tm = 14.9 min).
introduced dropwise and stirring was maintained (À20 °C,
10 min). Et3N (100 L) was introduced and the mixture was fil-
l
tered and evaporated. The residue was subjected to workup
(EtOAc) and flash chromatography (10?40% EtOAc–hexane) to
yield a fraction presumed to contain the disaccharide product as
a
pale yellow oil (130 mg). This residue was co-evaporated
(2 Â 10 mL MeCN) and used in the next reaction without further
purification or characterization. (b) A sliver of sodium was added
to a stirred solution of the crude product from (a) (0.302 mmol,
max.) in MeOH–THF (2:1, 3 mL) at 0 °C. The solution was allowed
to warm to rt and stirred (3 h). The mixture was neutralized
(Bio-Rad AGÒ-50W-X8 resin, H+ form), filtered and the resin was
washed with MeOH. The combined filtrate and washings were
evaporated to yield the alcohol as a colorless oil (98 mg). This res-
idue was co-evaporated (2 Â 10 mL MeCN) and used in the next
reaction without further purification or characterization. (c) The
crude product (0.302 mmol, max.) was dissolved in CH2Cl2 (3 mL)
and treated with benzoyl chloride (1.3 equiv per hydroxyl) and
pyridine (1 mL). The mixture was stirred at rt overnight, washed
with ice-chilled 0.5 M HCl and sat. NaHCO3. The organic phase
was dried (MgSO4), filtered, and evaporated, and the residue was
purified by flash chromatography (10?30% EtOAc–hexane) to give,
1.6. 2-Deoxy-2-sulfamino-
a-
D-glucopyranosyl-(1?4)-1,6-
anhydro-2-deoxy-2-sulfamino–
D
-glucopyranoside, disodium
salt (17)
A mixture of benzyl ether 16 (12.9 mg, 20.8 lmol) and Pearl-
firstly, the
a
-linked disaccharide 13 as a colorless foam (101 mg,
man’s catalyst (5 mg) in purified water (2 mL) was subjected to
50 psi H2 overnight. The mixture was filtered and lyophilized to
yield 10.7 mg (98%) of tetrol 17. 1H NMR (400 MHz, D2O) d 5.47
(br s, 1H, H-1I), 5.20 (d, 1H, J1-2 = 3.5, H-1II), 4.68 (br d, 1H, J5-
4 = 5.5, H-5), 4.07 (d, 1H, Jgem = 7.6, H-6AI), 3.98 (br s, 1H, H-3I),
3.75–3.64 (m, 4H), 3.52 (t, 1H, J2-3ꢀ3-4 = 9.3, H-3II), 3.34 (t, 1H, J3-
4ꢀ4-5 = 9.3, H-4II), 3.13 (obsd dd [partially obscured by H2I], 1H,
H-2II), 3.11 (br s, 1H, H-2I). Purity was 99% by CE (tm = 12.6 min).
46%, three steps). 1H NMR (400 MHz, CDCl3) d 3.11 (s, 1H; H-2I),
3.41 (dd, 1H, J1,2 = 3.7, J2,3 = 10.7 Hz; H-6I), 3.61 (s, 1H; H-3I), 3.39
(s, 1H; H-4I), 4.05 (d, 1H, J6,6 = 7.3 Hz; H-6I), 4.41–4.49 (m, 2H; H-
6II), 4.55, 4.68 (AB quartet, JA,B = 11.9 Hz; CH2Ph), 4.79 (ddd, 1H,
J4,5 = 10.3, J5,6 = 2.9, 5.9 Hz; H-5II), 4.91 (br d, 1H, J5,6 = 5.5 Hz; H-
5I), 5.08 (d, 1H, J1,2 = 3.6 Hz; H-1II), 5.51 (dd, 1H, J3,4 = 9.5,
J4,5 = 10.2 Hz, H-4II), 5.60 (s, 1H,; H-1I), 6.10 (dd, 1H, J2,3 = 10.7,
J3,4 = 9.3 Hz; H-3II), 7.29–7.55, 7.89–8.03 (2m, 20H; ArH); 13C
NMR (100 MHz, CDCl3) d 58.7, 61.5, 63.3, 64.8, 69.2, 69.5, 70.5,
73.2, 74.6, 78.1, 79.6, 100.7, 101.2, 128.1, 128.4, 128.5(8),
128.6(1), 128.6(4), 128.8, 128.9, 129.2, 129.8(6), 129.9(0), 130.1,
130.2, 133.4, 133.5, 133.7, 137.2, 165.6(1), 165.6(2), 166.3. Next,
the b-anomer 13b was obtained as a colorless oil (27 mg, 12%,
three steps). 1H NMR (400 MHz, CDCl3) d 3.19 (s, 1H; H-2I), 3.74
(dd, 1H, J5,6 = 6.2, J6,6 = 7.1 Hz; H-6I), 3.79–3.88 (m, 2H; H-2II, H-
3I), 3.88 (ddd, J4,5 = 9.2, J5,6 = 3.1, 4.7 Hz; H-5II), 3.95 (br s, 1H; H-
4I), 4.10 (d, 1H, J6,6 = 7.3 Hz; H-6I), 4.34 (dd, 1H; J5,6 = .9,
J6,6 = 12.2 Hz; H-6II), 4.50 (dd, 1H, J5,6 = 3.1, J6,6 = 12.3 Hz, H-6II),
4.52, 4.59 (AB quartet, JA,B 12.0 = Hz; CH2Ph), 4.65 (d, 1H,
J1,2 = 7.9 Hz; H-1II), 4.69 (br d, 1H, J5,6 = 5.5 Hz; H-5I), 5.44 (t, 1H,
J2,3=3,4 = 9.7 Hz; H-3II), 5.49 (br s, 1H; H-1I), 5.51 (t, 1H,
J3,4=4,5 = 9.6 Hz; H-4II), 7.23–7.50, 7.84–7.96 (2m, 20H; ArH).
Acknowledgments
We thank Drs. Siska Cochran, Cai Ping Li, Robert Don, and Ed-
ward Hammond for useful discussions.
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.carres.2009.09.012.
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