Synthesis of potential metal-binding group compounds to examine the zinc dependency of the GPI de-N-acetylase metalloenzyme in Trypanosoma brucei

Article abstract of DOI:10.1016/j.carres.2011.02.004

A small zinc-binding group (ZBG) library of deoxy-2-C-branched- monosaccharides, for example, 1,5-anhydroglucitols, consisting of either monodentate ligand binding carboxylic acids or bidentate ligand binding hydroxamic acids, were prepared to assess the

Full text of DOI:10.1016/j.carres.2011.02.004

Carbohydrate Research 346 (2011) 708–714
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Carbohydrate Research
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Synthesis of potential metal-binding group compounds to examine
the zinc dependency of the GPI de-N-acetylase metalloenzyme in
Trypanosoma brucei
⇑
Nuha Z. Abdelwahab, Michael D. Urbaniak, Michael A. J. Ferguson , Arthur T. Crossman
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, The University of Dundee, DD1 5EH Dundee, Scotland, United Kingdom
a r t i c l e i n f o
a b s t r a c t
Article history:
A small zinc-binding group (ZBG) library of deoxy-2-C-branched-monosaccharides, for example, 1,5-
anhydroglucitols, consisting of either monodentate ligand binding carboxylic acids or bidentate ligand
binding hydroxamic acids, were prepared to assess the zinc affinity of the putative metalloenzyme 2-
Received 15 December 2010
Received in revised form 1 February 2011
Accepted 2 February 2011
acetamido-2-deoxy-a-D-glucopyranosyl-(1?6)-phosphatidylinositol de-N-acetylase (EC 3.5.1.89) of gly-
Available online 24 February 2011
cosylphosphatidylinositol biosynthesis. The N-ureido thioglucoside was also synthesised and added to
the ZBG library because a previous N-ureido analogue, synthesised by us, had inhibitory activity against
the aforementioned de-N-acetylase, presumably via the N-ureido motif.
Keywords:
Glycosylphosphatidylinositol (GPI)
biosynthesis
Ó 2011 Elsevier Ltd. All rights reserved.
Zinc metalloenzyme inhibitor
Zinc-binding group
Branched monosaccharides,
Phosphatidylinositol de-N-acetylase
1. Introduction
we have designed and synthesised a small library of deoxymono-
saccharides [5–12 (Fig. 2)] containing recognisable zinc binding
Glycosylphosphatidylinositol (GPI) acts as a membrane anchor for
small but significant proportion of higher eukaryote cell-
groups (ZBGs), that is, carboxylic acids and hydroxamic acids, as
well as a potentially new ZBG, the ureido derivative, that should
a
surface glycoproteins that are particularly abundant in protozoan
parasites such as Trypanosoma brucei, the causative agent of African
sleeping sickness in humans and the related disease Nagana in cat-
tle.1 The structure, biosynthesis, and function of GPI anchors and re-
lated molecules have been extensively reviewed.1–4 Disruption of GPI
biosynthesis in the clinically relevant bloodstream form of T. brucei
has been genetically5–8 and chemically9 validated as a drug target.
A key early step in the biosynthesis of the GPI anchors is the
continue to probe the trypanosomal
acetylase.
a-D-GlcpNAc-PI de-N-
A good starting point for our compound library was the earlier
work by Hindsgaul and co-workers18,19 which demonstrated the
effectiveness of 1,5-anhydro-2-deoxy-D-glucitol hydroxamic acids,
for example 7,19 as ZBG probes. The hydroxamic acid 7 was resyn-
thesised and included in the compound library because 7 was
shown to be a potent inhibitor of LpxC,19 presumably via zinc che-
lation, and could serve as the standard by which to compare the
potency of the other analogues in the library. Therefore, com-
pounds 5, 6 and 8 resemble those of Hindsgaul et al. whereby the
2-C appendage is either a hydroxamic acid or a carboxylic acid
ZBG moiety. Compounds 9–11 were synthesised to supply potential
glycosyl donors for another project but might also exhibit some de-
gree of inhibition towards the trypanosome de-N-acetylase enzyme.
Lastly, the N-ureido thioglycoside 12 was fashioned because of pre-
vious inhibitory data of the N-ureido-GlcNAc-PI derivative 420
(Fig. 1) against the trypanosome de-N-acetylase enzyme. Analogue
12 is a truncated version of 4 which focuses on, what we believe
to be the most potent inhibitory component of 4, the N-ureido motif.
de-N-acetylation of 2-acetamido-2-deoxy-
(1?6)-phosphatidylinositol10
-GlcpNAc-PI (1, Fig. 1)] to form
-GlcpNH2-PI (2, Fig. 1). De-N-acetylation is a prerequisite for
a-D-glucopyranosyl-
[a-D
a-D
subsequent processing of 2 that leads to mature GPI anchor precur-
sors.11 In T. brucei, de-N-acetylation is followed by mannosylation
and subsequent inositol-acylation of 2, whereas in mammalian
cells the order of these reactions is reversed.12,13
Previously, we have shown14 that mammalian and trypanoso-
mal a-D-GlcpNAc-PI de-N-acetylases are zinc metalloenzymes, pro-
posed a mechanism of action similar to that of zinc peptidases and
postulated that known zinc binding motifs15,16 such as the
N-hydroxyurea analogue 3 (Fig. 1),17 could act as inhibitors. Here,
⇑
Corresponding author. Tel.: +44 1382 384219.
E-mail address: m.a.j.ferguson@dundee.ac.uk (M.A.J. Ferguson).
Compound 8 is in the literature, Jackman, J. E. et al. J. Biol. Chem. 2000, 275,
11002–11009, however, no preparative details are given therein.
0008-6215/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2011.02.004

N. Z. Abdelwahab et al. / Carbohydrate Research 346 (2011) 708–714
709
analytical data of 16). The benzyloxyamide 16 was hydrogenated, as
described in the literature, to give the hydroxamic acid 7;19 1H NMR
assignments for 7 were identical to those reported in the literature19
and see the Supplementary data for the 13C NMR assignments of 7.
The ZBG analogue 8 was synthesised following the sequence
16?1718?8, as previously described for 6. An alternative synthesis
of the derivative 1718 is described in the Supplementary data.
The synthesis of the targeted carboxylic acid 9 (Scheme 2) be-
gan from the acetolysis of the 1,6-anhydro derivative 1822 to give,
OH
R1
O
HO
HO
HO
OH
NH
OH
OH
O
O
O
P
OR2
O
R2
OCOC19H31
OCOC17H35
R1
exclusively, the
a-2-C-allyl derivative 19 [J1,2 = 3.1 Hz]. The tetra-
acetate derivative 19 proved to be a very useful intermediate be-
cause 19 could be altered to supply analogues 10 and 11, as well.
Thus, a portion of the 2-C-allyl intermediate 19 was ozonised to
give the aldehyde 20, which was oxidised, following Pinnicks’ pro-
tocols,21 to furnish the carboxylic acid 21 in 94% yield. Lastly, the
tetraacetate 21 was de-O-acetylated with 0.03 M methanolic so-
dium methoxide to produce the fully deprotected carboxylic acid
1
Ac
H
H2C
H2C
OCOC19H31
OCOC17H35
2
3
analogue 9 in 51% yield, as a mixture of
Another portion of the 2-C-allyl derivative 19 was transformed
into the corresponding - and b-phenylthioglucosides 22 and 23,
respectively, via Lewis acid (BF3ꢀEt2O) catalysed substitution of
the anomeric acetate with thiophenol in refluxing dichlorometh-
ane.23 These two anomers were separated by radial band chroma-
a/b anomers.
CONHOH
CONH2
C18H37
a
OCOC15H31
OCOC15H31
4
H2C
Figure 1.
tography to furnish the
a-anomer 22 (J1,2 = 4.9 Hz) and the
b-anomer 23 (J1,2 = 10.9 Hz) in 48% and 13% yields, respectively.
The closing sequences 22?24?26?10 and 23?25?27?11 were
then conducted without incident, essentially as those described for
9; the exception being 26?10 which was achieved via acid hydro-
lysis24 (Scheme 2).
2. Results and discussion
The synthesis, of the analogues 5–8, is based on a successful ap-
proach18,19 used previously (Scheme 1).
A synthesis of 1-thiophenyl-2-deoxy-2-ureido-b-D-glucopyran-
The first three steps, benzoylation?ozonolysis?Pinnick21 oxi-
dation, from the known18 2-C-allyl derivative 13 was accomplished
straightforwardly to furnish the pivotal carboxylic acid 14.19 The
carboxylic acid analogue 1419 and the corresponding intermediates
from 1318 were not fully characterised in the literature. Conse-
quently, we have included the analytical data for those intermedi-
ates, and that of compound 14,19 in this paper as Supplementary
data. Hydrogenolysis of the benzylidene protecting group of com-
pound 14 furnished the target analogue 5 in 59% yield; alterna-
tively, the yield could be improved to 70% by using aqueous TFA.
The synthesis of carboxylic acid 6 emerged from the de-O-ben-
zoylation of 14,19 under Zemplén conditions, followed by hydrog-
enolysis over 10% palladium on carbon to give the crude
derivative 6 (Scheme 1). The analogue 6 was then purified by
reversed phase chromatography (RPC) to afford the final target
glucitol 6 in 80% yield.
oside 12 was obtained on treatment of the known amine25 28 with
potassium cyanate (KOCN) and water at room temperature in total
darkness26,27 (Scheme 3). After evaporation to dryness, the crude
ureido compound was purified by reversed phase chromatography
to give crystalline 12 (65% yield; characteristic 13C carbonyl carbon
at d 158.47 ppm).
Details of the results of enzymatic studies with the above ZBG
analogues will be reported elsewhere in due course.
3. Experimental
3.1. General methods
1H, 13C, 31P NMR spectra were recorded on a Bruker AVANCE
500 MHz spectrometer using deuteriochloroform as a solvent and
tetramethylsilane as the internal standard, unless otherwise
indicated. All coupling constants (J) are given in Hertz. High reso-
lution electrospray ionisation mass spectra (HRESIMS) and liquid
chromatography mass spectra (LCMS) were recorded with a Bruker
The carboxylic acid derivative 1419 was coupled with O-ben-
zylhydroxylamine hydrochloride (BnONH2ꢀHCl) using N-(3-dimeth-
ylaminopropyl)-N0-ethylcarbodiimide hydrochloride (EDAC) to give
the known19 hydroxyamide 16 (see the Supplementary data for the
OH
OH
OH
OH
O
O
O
O
HO
BzO
HO
HO
HO
HO
HO
BzO
O
O
O
O
OH
O
OH
O
NHOH
O
NHOH
O
5
6
7
8
OH
OH
OH
OH
HO
HO
HO
HO
HO
HO
HO
HO
SPh
SPh
NH
OH
O
O
SPh
O
O
OH
OH
OH
NH2
9
10
11
12
Figure 2. A small library of zinc chelator probes.

710
N. Z. Abdelwahab et al. / Carbohydrate Research 346 (2011) 708–714
O
O
HO
O
O
BzO
Ph
Ph
O
O
3 Steps
O
OH
13
14
H2/ Pd/C/ AcOH
59%
NaOMe/ MeOH
75%
BnONH2 HCl/ EDAC
Et3N/ CH2Cl2
78%
(or) TFA(aq)/ THF
70%
O
O
HO
O
O
BzO
OH
Ph
Ph
O
O
O
HO
BzO
O
O
O
OH
NHOBn
OH
15
16
5
H2/ Pd/C/ AcOH
80%
H2/ Pd/C/ AcOH
67%
NaOMe/ MeOH
73%
OH
O
HO
HO
OH
O
O
Ph
O
O
HO
BzO
O
HO
OH
O
6
O
NHOBn
NHOH
17
7
H2/ Pd/C/ AcOH
62%
OH
O
HO
HO
O
NHOH
8
Scheme 1.
microTof spectrometer. Melting points were determined on a Reic-
hert hot-plate apparatus and are uncorrected. Optical rotations
were measured with a Perkin–Elmer 343 polarimeter. Thin layer
chromatography (TLC) was performed on Kieselgel 60 F254 (Merck)
or RP-18 F254s (Merck) plates with various solvent systems as
developers, followed by detection under UV light or by charring
using either sulfuric acid–water–ethanol (15:85:5), phospho-
molybdic acid, orcinol or ninhydrin spray reagents. Flash column
chromatography (FCC) was performed on Kieselgel 60 (0.040–
0.063 mm) (Merck). Reversed phase chromatography was per-
formed on a C18 cartridge supplied by Sigma–Aldrich. Radial-band
chromatography (RBC) was performed using a Chromatotron
(model 7924T, TC Research UK) with silica gel F254 TLC standard
grade as the adsorbent. All reactions were carried out in commer-
cially available dry solvents, unless otherwise stated. Light petro-
leum refers to the fraction having a boiling range 60–80 °C,
unless indicated otherwise.
pressure. The residue was purified by FCC (10:1:0.02 CHCl3–
MeOH–AcOH) to furnish a brown paste 5 (9 mg, 59%), which was
indistinguishable from that obtained by the following procedure.
3.2.1.2. Method B.
A solution of the benzylidene compound
1419 (40 mg, 0.10 mmol) in THF (2 mL) and 96% (aq) TFA (0.5 mL)
was stirred at room temperature for 3 h. The reaction mixture
was concentrated under reduced pressure and co-evaporated with
toluene (2 ꢁ 5 mL). The residue was purified with the same solvent
system as in method A to give the acid 5 (21 mg, 70%): Rf 0.20
(10:1:0.02 CHCl3–MeOH–AcOH); ½a D25
ꢂ
+8.9 (c 1.0, MeOH); 1H
NMR (CD3OD, 500 MHz): d 8.10–7.49 (m, 5H, Ph), 5.10 (dd, 1H,
J2,3 10.7, J3,4 9.3 Hz, H-3), 4.10 (dd, 1H, J1a,2 4.7, J1a,1b 11.5 Hz,
H-1a), 3.88 (dd, 1H, J5,6a 2.1, J6a,6b 11.8 Hz, H-6a), 3.71 (dd, 1H,
H-6b), 3.60 (t, 1H, J4,5 9.3 Hz, H-4), 3.40 (t, 1H, J1a,1b 11.5 Hz, H-
1b), 3.37–3.34 (m, 1H, H-5), 2.50–2.41 (m, 1H, H-2), 2.35 (dd, 1H,
J2,7a 4.8, J7a,7b 16.0 Hz, H-7a), 2.17 (dd, 1H, H-7b); 13C NMR (CD3OD,
125 MHz): d 175.49 (C@O), 168.15 (PhCO), 134.29–129.56 (C-Ph),
82.75 (C-5), 79.92 (C-3), 70.79 (C-4), 70.56 (C-1), 62.94 (C-6),
40.03 (C-2), 34.10 (C-7). HRESIMS: Calcd for [C15H18O7ꢃH]ꢃ:
309.0980. Found m/z: 309.0967.
3.2. Synthesis of the ZBG library
3.2.1. 1,5-Anhydro-3-O-benzoyl-2-C-carboxymethyl-2-deoxy-
D-glucitol (5)
3.2.1.1. Method A.
A solution of the benzylidene compound
3.2.2. 1,5-Anhydro-4,6-O-benzylidene-2-C-carboxymethyl-2-
deoxy-D-glucitol (15)
A methanolic 0.03 M MaOMe (0.6 mL, 0.018 mmol) solution
was added to the benzoate derivative 1419 (60 mg, 0.15 mmol) in
THF–MeOH (1:4 5 mL) and the reaction mixture was stirred over-
1419 (20 mg, 0.05 mmol) in AcOH (2 mL) containing 10% palladium
on carbon (10 mg) was stirred under a slight overpressure of
hydrogen at room temperature for 4 h. The reaction mixture was
filtered through a pad of Celite and concentrated under reduced

N. Z. Abdelwahab et al. / Carbohydrate Research 346 (2011) 708–714
711
O
OAc
OAc
Ac2O/ TFA
98%
O
O
AcO
AcO
OAc
OAc
18
19
PhSH/ BF3 Et2O/CH2Cl2/Δ
O3/ Ph3P/ CH2Cl2
88%
87%
OAc
OAc
O
O
AcO
AcO
AcO
R1
AcO
O
OAc
R2
H
1
2
20
22:
23:
R = H, R = SPh (48%)
R = SPh, R2 = H (13%)
1
NaClO2/ NaH2PO4
tert-BuOH/ Amylene/ H2O
O3/ Ph3P (or Me2S)
CH2Cl2
94%
OAc
O
OAc
O
AcO
AcO
AcO
AcO
R1
O
R2
O
OAc
H
OH
1
2
21
24:
R = H, R = SPh (88%)
25: R1 = SPh, R2 = H (87%)
NaClO2/ NaH2PO4
tert-BuOH/ Amylene/ H2O
NaOMe/ MeOH
51%
OH
O
OAc
HO
O
AcO
AcO
HO
R1
O
OH
O
R2
OH
9
OH
R = H, R2 = SPh (93%)
1
26:
27:
1
2
R = SPh, R = H (92%)
Δ
CH3COCH3/HCl/H2O/ for 26
10
and
NaOMe/ MeOH for 27
11
OH
O
HO
HO
R1
R2
O
OH
1
2
10:
11:
R = H, R = SPh (75%)
R = SPh, R2 = H (81%)
1
Scheme 2.
OH
OH
O
O
KOCN/ H2O/RT/darkness
65%
HO
HO
HO
HO
SPh
SPh
NH
NH2
O
28
NH2
12
Scheme 3.

712
N. Z. Abdelwahab et al. / Carbohydrate Research 346 (2011) 708–714
night at room temperature. Afterwards, the reaction mixture was
neutralised with Amberlite IR-120 (H+) ion-exchange resin, filtered
and the filtrate concentrated under reduced pressure and co-evap-
orated with water (5 ꢁ 5 mL). The residue was purified by FCC
(20:1:0.02 CH2Cl2–MeOH–AcOH) to give the crystalline acid 15
(33 mg, 75%): mp 183–185 °C; Rf 0.24 (20:1:0.02 CH2Cl2–MeOH–
J2,3 = J3,4 = 10.8 Hz, H-3), 5.01–4.91 (m, 3H, H-4, H-9a,b), 4.23 (dd,
1H, J5,6a 4.0, J6a,6b 12.4 Hz, H-6a), 3.97–3.93 (m, 2H, H-5, H-6b),
2.16–2.12 (m, 2H, H-2, H-7a), 2.09, 2.00, 1.97, 1.96 (4 ꢁ s, 12H,
4 ꢁ CH3CO), 1.97–1.93 (m, 1H, H-7b); 13C NMR (CDCl3,
125 MHz): d 169.78, 169.60, 168.83, 167.96 (4 ꢁ C@O), 133.06
(C-8), 116.54 (C-9), 90.68 (C-1), 70.81 (C-3), 68.73 (C-5), 68.09
(C-4), 60.90 (C-6), 41.89 (C-2), 30.71 (C-7), 19.82, 19.76, 19.70,
19.53, (4 ꢁ CH3CO). HRESIMS: Calcd for [C17H24O9+Na]+:
395.1313. Found m/z: 395.1298.
AcOH); ½a 2D5
ꢂ
ꢃ209.0 (c 1.0, MeOH); 1H NMR (CD3OD, 500 MHz): d
7.50–7.33 (m, 5H, Ph), 5.58 (s, 1H, PhCH), 4.20 (dd, 1H, J5,6a 5.0,
J6a,6b 10.3 Hz, H-6a), 4.02 (dd, 1H, J1a,2 4.7, J1a,1b 11.4 Hz, H-1a),
3.70 (t, 1H, J6a,6b 10.3 Hz, H-6b), 3.50–3.45 (m, 2H, H-3, H-4),
3.37–3.28 (m, 2H, H-1b, H-5), 2.77 (dd, 1H, J2,7a 3.0, J7a,7b 15.8 Hz,
H-7a), 2.21–2.14 (m, 1H, H-2), 2.11 (dd, 1H, H-7b); 13C NMR
3.2.6. 1,3,4,6-Tetra-O-acetyl-2-deoxy-2-C-formylmethyl-a-D-
glucopyranose (20)
(CD3OD, 125 MHz):
d
176.06 (C@O), 139.32–127.56 (C-Ph),
Ozone was passed through a solution of the allyl compound 19
(150 mg, 0.403 mmol) in CH2Cl2 (20 mL) at ꢃ78 °C until the solu-
tion turned blue. The excess ozone was removed by a stream of
argon until the solution was clear and then followed by the addi-
tion of triphenylphosphine (264.3 mg, 1.01 mmol). The mixture
was allowed to warm to room temperature for 2 h, concentrated
under reduced pressure and purified by RBC (6:1?2:1 light petro-
leum–EtOAc) to give the aldehyde 20 (84 mg, 88%): Rf 0.28 (1:1
103.06 (PhCH), 84.55 (C-4), 73.57 (C-3), 73.17 (C-5), 71.46 (C-1),
69.84 (C-6), 41.88 (C-2), 33.58 (C-7). HRESIMS: Calcd for
[C15H18O6ꢃH]ꢃ: 293.1031. Found m/z: 293.1030.
3.2.3. 1,5-Anhydro-2-C-carboxymethyl-2-deoxy-D-glucitol (6)
A solution of the benzylidene derivative 15 (58 mg, 0.20 mmol)
in AcOH (10 mL) containing 10% palladium on carbon (29 mg) was
stirred under a slight overpressure of hydrogen at room tempera-
ture for 3 h. The reaction mixture was filtered through a pad of Cel-
ite and concentrated under reduced pressure. The resulting residue
was purified by an RPC C18 column (10% MeOH) to furnish the car-
light petroleum–EtOAc); ½a D25
ꢂ
+172.7 (c 0.6, CHCl3); 1H NMR
(CDCl3, 500 MHz): d 9.61 (t, 1H, J 1.2 Hz, HC@O), 6.20 (d, 1H, J1,2
3.4 Hz, H-1), 5.18 (dd, 1H, J2,3 11.4, J3,4 9.5 Hz, H-3), 5.03 (t, 1H,
J4,5 9.5 Hz, H-4), 4.23 (dd, 1H, J5,6a 4.6, J6a,6b 13.0 Hz, H-6a),
4.01–3.97 (m, 2H, H-5, H-6b), 2.76–2.70 (m, 1H, H-2), 2.38 (m,
2H, H-7a, H-7b), 2.09, 2.02, 19.96, 19.94 (4 ꢁ s, 12H, 4 ꢁ CH3CO);
13C NMR (CDCl3, 125 MHz): d 197.80 (HC@O), 169.70, 169.52,
168.66, 167.88 (4 ꢁ C@O), 90.86 (C-1), 70.18 (C-3), 68.84 (C-5),
67.80 (C-4), 60.73 (C-6), 41.42 (C-7), 36.92 (C-2), 19.85, 19.80,
19.70, 19.62, (4 ꢁ CH3CO). HRESIMS: Calcd for [C16H22O10+Na]+:
397.1313. Found m/z: 397.1298.
boxylic acid 6 (32 mg, 80%): Rf 0.40 (10% MeOH); ½a D25
ꢂ
+379.6 (c 0.5,
MeOH); 1H NMR (CD3OD, 500 MHz): d 3.99 (dd, 1H, J1a,2 3.9, J1a,1b
11.5 Hz, H-1a), 3.83 (dd, 1H, J5,6a 2.1, J6a,6b 11.8 Hz, H-6a), 3.62
(dd, 1H, H-6b), 3.23 (t, 1H, J3,4 = J4,5 = 8.6 Hz, H-4), 3.19–3.15 (m,
3H, H-1b, H-3, H-5), 2.77–2.71 (m, 1H, H-7a), 2.08–2.00 (m, 2H,
H-2, H-7b); 13C NMR (CD3OD, 125 MHz): d 176.44 (C@O), 82.71
(C-5), 77.59 (C-3), 73.12 (C-4), 70.81 (C-1), 63.26 (C-6), 41.34 (C-
2), 33.83 (C-7). HRESIMS: Calcd for [C8H14O6ꢃH]ꢃ: 205.0718.
Found m/z: 205.0724.
3.2.7. 1,3,4,6-Tetra-O-acetyl-2-C-carboxymethyl-2-deoxy-a-D-
glucopyranose (21)
3.2.4. 1,5-Anhydro-2-C-(carboxymethyl N-hydroxyamide)-2-
deoxy-D-glucitol (8)
A solution of sodium chlorite (2.58 g, 28.56 mmol) and sodium
dihydrogen phosphate (3.92 g, 32.63 mmol) in water (20 mL) was
10% Palladium on carbon (40 mg) was added to a solution of the
benzyloxyamide 1718 (40 mg, 0.10 mmol) in AcOH (10 mL). The
reaction mixture was stirred under a slight over pressure of hydro-
gen at room temperature for 4 h. After filtration through a pad of
Celite the solvent was concentrated under reduced pressure. The
resulting residue was purified by an RPC C18 column (10% MeOH)
to furnish the hydroxamic acid 8 (13.6 mg, 62%): Rf 0.38 (10%
added dropwise to a solution of the aldehyde 20 (724 mg,
1.931 mmol) in tert-BuOH (56.7 mL, 604 mmol) and amylene
(17 mL, 203 mmol). The reaction mixture was stirred for 1 h then
diluted with ice water and extracted with EtOAc (2 ꢁ 50 mL). The
combined organic extracts were washed with brine (30 mL), dried
(Na2SO4), and concentrated under reduced pressure. The residue
was purified by FCC (1:1:0.02 light petroleum–EtOAc–AcOH) to
furnish the acid 21 (709 mg, 94%): Rf 0.27 (1:1:0.02 light petro-
MeOH); ½a 2D5
ꢂ
+38.9 (c 1.3, MeOH); 1H NMR (CD3OD, 500 MHz): d
3.92 (dd, 1H, J1a,2 4.6, J1a,1b 11.5 Hz, H-1a), 3.83 (dd, 1H, J5,6a 1.9,
J6a,6b 11.8 Hz, H-6a), 3.61 (dd, 1H, H-6b), 3.24–3.13 (m, 4H, H-1b,
H-3, H-4, H-5), 2.54 (dd, 1H, J2,7a 3.9, J7a,7b 14.3 Hz, H-7a), 2.06–
1.95 (m, 1H, H-2), 1.84 (dd, 1H, H-7b); 13C NMR (CD3OD,
125 MHz): d 171.44 (C@O), 82.72, 77.99, 73.08, 70.66 (C-1), 63.26
(C-6), 41.64 (C-2), 32.66 (C-7). HRESIMS: Calcd for [C8H15NO6+-
Na]+: 244.0792. Found m/z: 244.0795.
leum–EtOAc–AcOH); ½a D25
ꢂ
+88.3 (c 1.3, CHCl3); 1H NMR (CDCl3,
500 MHz): d 6.25 (d, 1H, J1,2 3.4 Hz, H-1), 5.22 (dd, 1H, J2,3 11.4,
J3,4 9.5 Hz, H-3), 5.03 (t, 1H, J4,5 9.5 Hz, H-4), 4.28 (dd, 1H, J5,6a
4.1, J6a,6b 12.4 Hz, H-6a), 4.08–4.02 (m, 2H, H-5, H-6b), 2.61–2.56
(m, 1H, H-2), 2.34 (dd, 1H, J2,7a 5.8, J7a,7b 16.3 Hz, H-7a), 2.27 (dd,
1H, H-7b), 2.04, 2.01, 2.00, 19.99 (4 ꢁ s, 12H, 4 ꢁ CH3CO); 13C
NMR (CDCl3, 125 MHz): d 174.59, 172.40, 172.14, 171.44, 170.75
(5 ꢁ C@O), 93.16 (C-1), 72.67 (C-3), 71.13 (C-5), 70.45 (C-4),
63.16 (C-6), 41.79 (C-2), 32.98 (C-7), 20.79, 20.75, 20.70, 20.65
(4 ꢁ CH3CO). HRESIMS: Calcd for [C16H22O11ꢃH]ꢃ: 389.1089.
Found m/z: 389.1085.
3.2.5. 1,3,4,6-Tetra-O-acetyl-2-C-allyl-2-deoxy-a-D-
glucopyranose (19)
A solution of the known22 1,6-anhydro derivative 18 (0.865 g,
3.2 mmol) in Ac2O–trifluoroacetic acid (9:1, 20 mL) was stirred at
room temperature overnight, whereafter it was neutralised with
a solution of satd NaHCO3. The aqueous solution was extracted
with CH2Cl2 (2 ꢁ 200 mL) and the organic extracts were combined,
washed with H2O (200 mL), brine (200 mL), dried with MgSO4 and
then concentrated under reduced pressure. The residue was puri-
fied by FCC (5:1?2:1 light petroleum–EtOAc) to give the tetraace-
tate 19 as white needles (1.17 g, 98%): mp 99–101 °C (from 10:1
3.2.8. 2-C-Carboxymethyl-2-deoxy-D-glucopyranose (9)
To a solution of benzoylated compound 21 (93 mg, 0.238 mmol)
in MeOH (2 mL) was added 0.03 M sodium methoxide in MeOH
(6.2 mL, 0.186 mmol) at room temperature. After 48 h, the reaction
mixture was neutralised with Amberlite IR-120 (H+) ion-exchange
resin, filtered and the filtrate was concentrated under reduced
pressure; followed by co-evaporation with water (5 ꢁ 5 mL). The
residue was purified by FCC (3:1:0.02 CH2Cl2–MeOH–AcOH) to
light petroleum–EtOH); Rf 0.20 (1:1 light petroleum–EtOAc); ½a D25
ꢂ
+123.0 (c 1.0, CHCl3); 1H NMR (CDCl3, 500 MHz): d 6.06 (d, 1H,
J1,2 3.1 Hz, H-1), 5.65–5.57 (m, 1H, H-8), 5.20 (t, 1H,
give the carboxylic acid 9 as an
a:b (1.5:1) mixture (27 mg, 51%):
Rf 0.25 (3:1:0.02 CH2Cl2–MeOH–AcOH); 1H NMR (CD3OD,

N. Z. Abdelwahab et al. / Carbohydrate Research 346 (2011) 708–714
713
500 MHz): d 5.23 (d, 1H, J1,2 3.1 Hz, H-1
1b), 3.85 (dd, J5,6a 1.9, J6a,6b 11.7 Hz, H-6ab), 3.80–3.76 (m, 2H, H-
5, H-6a ), 3.70 (dd, J6a,6b 11.4 Hz, H-6b ), 3.66 (dd, H-6bb), 3.51
(dd, J2,3 10.9, J3,4 8.9 Hz, H-3 ), 3.36 (dd, J2,3 10.7, J3,4 8.1 Hz, H-
3b), 3.32–3.21 (m, 3H, H-4 , H-4b, H-5b), 2.71 (dd, J2,7a 3.3, J7a,7b
16.6 Hz, H-7a ), 2.60 (dd, J2,7a 4.1, J7a,7b 16.1 Hz, H-7ab), 2.46 (dd,
H-7bb), 2.39 (dd, H-7b
1H, H-2b); 13C NMR (CD3OD, 125 MHz):
(2 ꢁ C@O), 98.23 (C-1b), 93.79 (C-1
73.25 (C-5 ), 73.17 (C-3 ), 72.90 (C-4), 63.06 (C-6b), 63.02 (C-
a
), 4.63 (d, J1,2 8.6 Hz, H-
(HC@O), 170.63, 170.31, 169.85 (3 ꢁ C@O), 132.96–127.85 (C-Ph),
87.52 (C-1), 71.92 (C-3), 69.54 (C-4), 68.62 (C-5), 62.24 (C-6),
43.37 (C-7), 39.7 3 (C-2), 20.72, 20.69, 20.67 (3 ꢁ CH3CO). HRE-
SIMS: Calcd for [C20H24O8S+Na]+: 447.1084. Found m/z: 447.1096.
a
a
a
a
a
3.2.11. Phenyl 3,4,6-tri-O-acetyl-2-deoxy-2-C-formylmethyl-1-
thio-b-D-glucopyranoside (25)
This compound was prepared from the allyl derivative 23
(25 mg, 0.059 mmol) essentially as described for the previous
a
), 2.08–2.03 (m, 1H, H-2
a
), 1.91–1.85 (m,
177.02, 175.55
d
a
), 77.99 (C-5b), 76.06 (C-3b),
a
a
a-derivative 24. However, dimethyl sulfide (130 lL, 0.177 mmol)
6
a
) 47.62 (C-2b), 44.83 (C-2
a
), 33.62 (C-7
a
), 33.51 (C-7b). HRE-
was used in place of triphenylphosphine. The residue was purified
by RBC (6:1?2:1 light petroleum–EtOAc) to afford the aldehyde 25
SIMS: Calcd for [C8H14O7ꢃH]ꢃ: 221.0667. Found m/z: 221.0659.
(21.8 mg, 87%): Rf 0.21 (2:1 light petroleum–EtOAc); ½a D25
ꢂ
+11.0
3.2.9. Phenyl 3,4,6-tri-O-acetyl-2-C-allyl-2-deoxy-1-thio-
b- -glucopyranoside (22) and (23)
To stirred solution of the tetraacetate 19 (200 mg,
0.537 mmol) in freshly distilled CH2Cl2 (10 mL) at room tempera-
ture under argon was added thiophenol (110 L, 1.074 mmol)
and boron trifluoride diethyl etherate (270 L, 2.148 mmol). The
resulting mixture was heated to reflux for 3 h, cooled to room tem-
perature, and then diluted with CH2Cl2 (10 mL), washed with satd
NaHCO3 (10 mL), brine (10 mL), dried (Na2SO4), and concentrated
under reduced pressure. RBC (10:1?4:1 light petroleum–EtOAc)
a- and
(c 1.5, CHCl3); 1H NMR (CDCl3, 500 MHz): d 9.59 (s, 1H, HC@O),
7.53–7.31 (m, 5H, Ph), 5.15 (dd, 1H, J2,3 10.7, J3,4 9.5 Hz, H-3),
4.96 (t, 1H, J4,5 9.5 Hz, H-4), 4.84 (d, 1H, J1,2 10.7 Hz, H-1), 4.27
(dd, 1H, J5,6a 5.3, J6a,6b 12.2 Hz, H-6a), 4.17 (dd, 1H, H-6b), 3.76–
3.71 (m, 1H, H-5), 2.84 (dd, 1H, J2,7a 3.8, J7a,7b 16.4 Hz, H-7a), 2.57
(dd, 1H, H-7b), 2.45–2.38 (m, 1H, H-2), 2.10, 2.01, 1.97 (3 ꢁ s, 9H,
D
a
l
l
3 ꢁ CH3CO); 13C NMR (CDCl3, 125 MHz):
d 198.94 (HC@O),
170.69, 170.31, 169.82 (3 ꢁ C@O), 132.87–128.45 (C-Ph), 86.51
(C-1), 75.67 (C-5), 74.49 (C-3), 69.26 (C-4), 62.47 (C-6), 43.09
(C-7), 40.51 (C-2), 20.81, 20.67, 20.61 (3 ꢁ CH3CO). HRESIMS: Calcd
for [C20H24O8S+Na]+: 447.1084. Found m/z: 447.1096.
of the residue provided the
a-anomer 22 (95 mg, 48%), the
b-anomer 23 (25.7 mg, 13%), as well as, an
a/b mixture (51.5 mg,
26%).
3.2.12. Phenyl 3,4,6-tri-O-acetyl-2-C-carboxymethyl-2-deoxy-1-
thio-a-D-glucopyranoside (26)
a
-Anomer 22: Rf 0.28 (4:1 light petroleum–EtOAc); ½a D25
ꢂ
+281.0
(c 1.0, CHCl3); 1H NMR (CDCl3, 500 MHz): d 7.47–7.27 (m, 5H, Ph)
5.76–5.68 (m, 1H, H-8), 5.46 (d, 1H, J1,2 4.9 Hz, H-1), 5.23–5.18
(m, 2H, H-3, H-9a), 5.08 (dd, 1H, J 9.8 Hz, H-9b), 5.00 (t, 1H,
J3,4 = J4,5 = 10.2 Hz, H-4), 4.63–4.60 (m, 1H, H-5), 4.31 (dd, 1H, J5,6a
5.2, J6a,6b 12.3 Hz, H-6a), 4.01 (dd, 1H, H-6b), 2.44–2.38 (m, 1H,
H-2), 2.32–2.26 (m, 1H, H-7a), 2.25–2.16 (m, 1H, H-7b) 2.05,
2.04, 2.03 (3 ꢁ s, 12H, 3 ꢁ CH3CO); 13C NMR (CDCl3, 125 MHz): d
170.63, 170.31, 169.97 (3 ꢁ C@O), 134.06 (C-8), 133.57–127.64
(C-Ph), 117.94 (C-9), 88.09 (C-1), 72.46 (C-3), 69.98 (C-4), 68.80
(C-5), 62.41 (C-6), 45.04 (C-2), 33.03 (C-7), 20.73, 20.72, 20.69
(3 ꢁ CH3CO). HRESIMS: Calcd for [C21H26O7S+Na]+: 445.1291.
Found m/z: 445.1294.
Pinnick21 oxidation of the aldehyde 24 (0.383 g, 0.902 mmol) in
the presence of sodium chlorite (1.20 g, 13.04 mmol), sodium dihy-
drogen phosphate (1.83 g, 15.24 mmol), tert-BuOH (26.5 mL,
282 mmol), amylene (7.96 mL, 94.71 mmol) and water (10 mL),
essentially as described for compound 21, furnished a crude resi-
due of 26. This residue was purified by FCC (1:1:0.02 hexane–
Et2O–AcOH) to give the white crystalline carboxylic acid 26
(0.369 g, 93%): mp 95–98 °C; Rf 0.25 (1:1:0.02 hexane–Et2O–
AcOH); ½a 2D5
ꢂ
+195.0 (c 1.0, CHCl3); 1H NMR (CDCl3, 500 MHz): d
7.46–7.27 (m, 5H, Ph), 5.77 (d, 1H, J1,2 5.1 Hz, H-1), 5.18 (dd, 1H,
J2,311.7, J3,4 9.1 Hz, H-3), 5.03 (dd, 1H, J4,5 10.1 Hz, H-4), 4.61–
4.56 (m, 1H, H-5), 4.32 (dd, 1H, J5,6a 5.1, J6a,6b 12.3 Hz, H-6a), 4.05
(dd, 1H, H-6b), 2.90–2.85 (m, 1H, H-2), 2.61 (dd, 1H, J2,7a 8.4,
J7a,7b 17.0 Hz, H-7a), 2.50 (dd, 1H, H-7b), 2.05, 2.04, 2.02 (3 ꢁ s,
9H, 3 ꢁ CH3CO); 13C NMR (CDCl3, 125 MHz): d 176.64, 170.86,
170.43, 170.03 (4 ꢁ C@O), 133.10–127.79 (C-Ph), 87.65 (C-1),
71.94 (C-3), 69.75 (C-4), 68.65 (C-5), 62.30 (C-6), 41.70 (C-2),
33.78 (C-7), 20.84, 20.70, 20.61 (3 ꢁ CH3CO). HRESIMS: Calcd for
[C20H24O9SꢃH]ꢃ: 439.1068. Found m/z: 439.1085.
b-Anomer 23: Rf 0.24 (4:1 light petroleum–EtOAc); ½a D25
ꢂ
+60.0 (c
0.2, CHCl3); 1H NMR (CDCl3, 500 MHz): d 7.56–7.31 (m, 5H, Ph)
5.80–5.71 (m, 1H, H-8), 5.14–5.08 (m, 3H, H-3, H-9a, H-9b), 4.94
(dd, 1H, J3,4 = J4,5 = 10.0 Hz, H-4), 4.55 (d, 1H, J1,2 10.9 Hz, H-1),
4.24 (dd, 1H, J5,6a 5.6, J6a,6b 12.1 Hz, H-6a), 4.13 (dd, 1H, H-6b),
3.65–3.60 (m, 1H, H-5), 2.45–2.40 (m, 1H, H-7a), 2.34–2.29 (m,
1H, H-7b), 2.07 (s, 3H, CH3CO), 2.06–2.02 (m, 1H, H-2), 2.01, 2.00
(2 ꢁ s, 6H, 2 ꢁ CH3CO); 13C NMR (CDCl3, 125 MHz): d 170.71,
170.29, 169.94 (3 ꢁ C@O), 132.47 (C-8), 132.79–128.11 (C-Ph),
118.93 (C-9), 86.45 (C-1), 75.32 (C-5), 73.16 (C-3), 69. 86 (C-4),
62.70 (C-6), 43.81 (C-2), 32.05 (C-7), 20.79, 20.75, 20.70
(3 ꢁ CH3CO).
3.2.13. Phenyl 3,4,6-tri-O-acetyl-2-C-carboxymethyl-2-deoxy-1-
thio-b-D-glucopyranoside (27)
Pinnick21 oxidation of the aldehyde 25 (20 mg, 0.047 mmol) in
the presence of sodium chlorite (63 mg, 0.695 mmol), sodium
dihydrogen phosphate (95 mg, 0.794 mmol), tert-BuOH (1.40 mL,
3.2.10. Phenyl 3,4,6-tri-O-acetyl-2-deoxy-2-C-formylmethyl-1-
14.71 mmol), amylene (413.5 lL, 4.94 mmol) and water (10 mL),
thio-
a
-
D
-glucopyranoside (24)
essentially as described for compound 21, gave the crude com-
pound 27. This material was purified by FCC (1:1:0.02 hexane–
Et2O–AcOH) and gave the acid 27 as white crystals (19.4 mg,
This compound was prepared from the allyl derivative 22
(95 mg, 0.225 mmol) and then quenched with triphenylphosphine
(147 mg, 0.562 mmol) essentially as described for 20. RBC
(6:1?2:1 light petroleum–EtOAc) of the residue yielded the alde-
92%): mp 95–98 °C; Rf 0.25 (1:1:0.02 hexane–Et2O–AcOH); ½a D25
ꢂ
+10 (c 1.0, CHCl3); 1H NMR (CDCl3, 500 MHz): d 7.50–7.31 (m,
5H, Ph), 5.26 (dd, 1H, J2,3 10.6 Hz, J3,4 9.3, H-3), 4.95 (t, 1H, J4,5
9.3 Hz, H-4), 4.92 (d, 1H, J1,2 10.9 Hz, H-1), 4.25 (dd, 1H, J5,6a 5.3,
J6a,6b 12.2 Hz, H-6a), 4.17 (dd, 1H, H-6b), 3.75–3.71 (m, 1H, H-5),
2.69 (dd, 1H, J2,7a 4.0, J7a,7b 17.1 Hz, H-7a), 2.61 (dd, 1H, H-7b),
2.35–2.28 (m, 1H, H-2), 2.09, 2.01, 1.99 (3 ꢁ s, 9H, 3 ꢁ CH3CO);
13C NMR (CDCl3, 125 MHz): d 176.26, 170.75, 170.49, 169.85
(4 ꢁ C@O), 132.96–128.37 (C-Ph), 86.24 (C-1), 75.59 (C-5), 73.97
(C-3), 69.43 (C-4), 62.53 (C-6), 41.93 (C-2), 32.94 (C-7), 20.81,
hyde 24 (84 mg, 88%): Rf 0.21 (2:1 light petroleum–EtOAc); ½a D25
ꢂ
+268.27 (c 0.9, CHCl3); 1H NMR (CDCl3, 500 MHz): d 9.74 (s, 1H,
HC@O), 7.44–7.28 (m, 5H, Ph), 5.75 (d, 1H, J1,2 5.1 Hz, H-1), 5.16
(dd, 1H, J2,3 11.5, J3,4 9.5 Hz, H-3), 5.04 (t, 1H, J4,5 9.5 Hz, H-4),
4.61–4.56 (m, 1H, H-5), 4.32 (dd, 1H, J5,6a 5.1, J6a,6b 12.3 Hz, H-
6a), 4.05 (dd, 1H, H-6b), 3.03–2.95 (m, 1H, H-2), 2.76 (dd, 1H,
J2,7a 8.1, J7a,7b 18.3 Hz, H-7a), 2.61 (dd, 1H, H-7b), 2.07, 2.04, 2.02
(3 ꢁ s, 9H, 3 ꢁ CH3CO); 13C NMR (CDCl3, 125 MHz): d 198.86

714
N. Z. Abdelwahab et al. / Carbohydrate Research 346 (2011) 708–714
20.70, 20.61 (3 ꢁ CH3CO). HRESIMS: Calcd for [C20H24O9SꢃH]ꢃ:
439.1068. Found m/z: 439.1085.
125.85 (C-Ph), 86.56 (C-1), 80.87 (C-5), 76.04 (C-3), 70.54 (C-4),
60.97 (C-6), 54.94 (C-2). HRESIMS: Calcd for [C13H18N2O5S+H]+:
315.1009. Found m/z: 315.1012.
3.2.14. Phenyl 2-C-carboxymethyl-2-deoxy-1-thio-a-D-
glucopyranoside (10)
Acknowledgements
To a stirred mixture of the triacetate 26 (75 mg, 0.170 mmol) in
acetone (10 mL) at 56 °C was added dropwise a solution of concen-
trated hydrochloric acid (1 mL) in water (1.8 mL). Stirring was con-
tinued overnight at 56 °C, whereafter the mixture was neutralised
with TEA, concentrated under reduced pressure and co-evaporated
with toluene (2 ꢁ 5 mL). The residue was purified by an RPC C18
column (55% MeOH) to furnish the carboxylic acid as white needles
This work was supported by a programme grant from The Well-
come Trust (085622). One of the authors (N. Z. Abdelwahab) is in-
debted to the BBSRC for a Ph.D. studentship. We also thank Dr. A. V.
Nikolaev for his interest and advice.
10 (40 mg, 75%): mp 144–146 °C; Rf 0.38 (55% MeOH); ½a D25
ꢂ
+247 (c
1.0, MeOH); 1H NMR (CD3OD, 500 MHz): d 7.49–7.26 (m, 5H, Ph),
5.61 (d, 1H, J1,2 4.5 Hz, H-1), 4.12–4.09 (m, 1H, H-5), 3.80 (dd,
1H, J6a,6b 12.0 Hz, H-6a), 3.76 (dd, 1H, J5,6b 4.9 Hz, H-6b), 3.42–
3.34 (m, 2H, H-3, H-4), 2.88 (dd, 1H, J2,7a 3.7, J7a,7b 16.0 Hz, H-7a),
2.56–2.46 (m, 2H, H-2, H-7b); 13C NMR (CD3OD, 125 MHz): d
174.48 (COOH), 136.11–128.54, (C-Ph), 90.54 (C-1), 75.13 (C-5),
74.09, 72.93, 62.56 (C-6), 45.29 (C-2), 34.85 (C-7). HRESIMS: Calcd
for [C14H18O6SꢃH]ꢃ: 313.0751. Found m/z: 313.0766.
Supplementary data
Supplementary data (additional experimental procedures and
partial characterisation data for those intermediates obtained from
the sequence 13?14, hydroxamates 16 and 17, plus the hydroxa-
mic acid 7) associated with this article can be found, in the online
version, at doi:10.1016/j.carres.2011.02.004.
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81%): mp 205–208 °C; Rf 0.42 (55% MeOH); ½a D25
ꢃ50.0 (c 0.6,
ꢂ
MeOH); 1H NMR (CD3OD, 500 MHz): d 7.54–7.27 (m, 5H, Ph),
4.85 (d, 1H, J1,2 10.7 Hz, H-1), 3.86 (dd, 1H, J6a,6b 11.9 Hz, H-6a),
3.68 (dd, 1H, J5,6b 5.4 Hz, H-6b), 3.51 (t, 1H, J2,3 = J3,4 = 8.9 Hz, H-
3), 3.34–3.29 (m, 1H, H-5), 3.26 (t, 1H, J4,5 8.9 Hz, H-4), 2.73 (dd,
1H, J2,7a 3.2, J7a,7b 16.6 Hz, H-7a), 2.60 (dd, 1H, H-7b), 2.04–1.96
(m, 1H, H-2); 13C NMR (CD3OD, 125 MHz): d 174.59 (COOH),
135.78–128.22 (C-Ph), 88.85 (C-1), 81.82 (C-5), 77.85 (C-3), 72.83
(C-4), 63.10 (C-6), 45.59 (C-2), 36.85 (C-7). HRESIMS: Calcd for
[C14H18O6SꢃH]ꢃ: 313.0751. Found m/z: 313.0766.
3.2.16. Phenyl 2-(N-aminocarbonyl)amino-2-deoxy-1-thio-b-D-
glucopyranoside (12)
Potassium cyanate (278 mg, 3.42 mmol) was added to a suspen-
sion of the known25 amino-glucopyranoside 28 (607 mg,
2.24 mmol) in water (15 mL). The mixture was stirred in total
darkness at room temperature for 4 days. Whereafter, the water
was evaporated to dryness under reduced pressure and the residue
was co-evaporated with toluene (3 ꢁ 20 mL). RPC (25% CH3CN) of
the residue yielded the ureido compound 12 (458 mg, 65%): mp
220–222 °C (MeOH); Rf 0.40 (25% CH3CN); ½a D25
ꢃ32.0 (c 1.5,
ꢂ
DMSO); 1H NMR (DMSO, 500 MHz): d 7.41–7.17 (m, 5H, Ph), 6.02
(d, 1H, J 8.7 Hz, NH), 5.52 (s, 2H, NH2), 5.07 (d, 2H, J 5.2 Hz, OH-3
and OH-4), 4.81 (d, 1H, J1,2 10.3 Hz, H-1), 4.62 (dd, 1H, J 5.8, J
11.5 Hz, 6-OH), 3.70 (ddd, 1H, J5,6a 5.3, J6a,6b 11.8 Hz, H-6a), 3.45
(d, 1H, H-6b), 3.40 (ddd, 1H, J2,3 9.0 Hz, H-2), 3.30 (dt, 1H, J3,4
9.0.Hz, H-3), 3.24–3.21 (m, 1H, H-5), 3.13 (dt, 1H, J4,5 9.0 Hz,
H-4); 13C NMR (DMSO, 125 MHz): d 158.47 (C@O), 136.17–
27. Baker, B. R.; Hullar, T. L. J. Org. Chem. 1965, 30, 4038–4044.