Synthesis of C-Glycosyl Phosphate and Phosphonate, Analogues of N-Acetyl-α-D-Glucosamine 1-Phosphate

Article abstract of DOI:10.1081/CAR-120026465

Synthesis of α-C-ethylene phosphate and phosphonate as well as α-C-methylene phosphate analogues of N-acetyl-α-D-glucosamine 1-phosphate is reported starting from the common perbenzylated 2-acetamido-2-deoxy-α-C-allyl glucoside. Anomerisation of the corre

Full text of DOI:10.1081/CAR-120026465

Journal of Carbohydrate Chemistry
ISSN: 0732-8303 (Print) 1532-2327 (Online) Journal homepage: http://www.tandfonline.com/loi/lcar20
Synthesis of C‐Glycosyl Phosphate
and Phosphonate, Analogues of
N‐Acetyl‐α‐d‐Glucosamine 1‐Phosphate
Olivier Gaurat , Juan Xie & Jean‐Marc Valéry
To cite this article: Olivier Gaurat , Juan Xie & Jean‐Marc Valéry (2003) Synthesis of C‐Glycosyl
Phosphate and Phosphonate, Analogues of N‐Acetyl‐α‐d‐Glucosamine 1‐Phosphate ,
Journal of Carbohydrate Chemistry, 22:7-8, 645-656, DOI: 10.1081/CAR-120026465
To link to this article: http://dx.doi.org/10.1081/CAR-120026465
Published online: 23 Feb 2007.
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Date: 04 October 2015, At: 22:06

JOURNAL OF CARBOHYDRATE CHEMISTRY
Vol. 22, Nos. 7 & 8, pp. 645–656, 2003
Synthesis of C-Glycosyl Phosphate and Phosphonate,
Analogues of N-Acetyl-a-D-Glucosamine 1-Phosphatey
*
Olivier Gaurat, Juan Xie, and Jean-Marc Vale´ry
Structure et Fonction de Mole´cules Bioactives, Equipe de Chimie des Glucides,
Universite´ Pierre et Marie Curie, Paris, France
ABSTRACT
Synthesis of a-C-ethylene phosphate and phosphonate as well as a-C-methylene
phosphate analogues of N-acetyl-a-D-glucosamine 1-phosphate is reported starting
from the common perbenzylated 2-acetamido-2-deoxy-a-C-allyl glucoside. Anom-
erisation of the corresponding amino a-C-glucosyl aldehyde to the b-aldehyde
was observed. Thus, both amino a- and b-C-glucosyl methanol were obtained
after reduction.
Key Words: Glycosyl phosphate; a-C-glucosyl phosphate; a-C-glucosyl
phosphonate; Amino C-glycosides.
INTRODUCTION
Glycosyl phosphates are essential precursors in the biosynthesis of the oligo-
saccharidic chains of glycoconjugates. The preparation of C-glycosyl phosphates and
phosphonates as metabolically stable mimics of natural glycosyl phosphates is of great
yThis paper is dedicated to Professor Ge´rard Descotes on the occasion of his 70th birthday.
*
Correspondence: Juan Xie, Laboratoire de Chimie des Glucides, Universite´ Pierre et Marie
Curie, CNRS UMR 7613, 4 place Jussieu, 75005 Paris, France; Fax: 33-1-44-27-55-13;
E-mail: xie@ccr.jussieu.fr.
645
DOI: 10.1081/CAR-120026465
0732-8303 (Print); 1532-2327 (Online)
www.dekker.com
Copyright D 2003 by Marcel Dekker, Inc.

646
Gaurat, Xie, and Vale´ry
Figure 1. Structure of GlcNAc-1-P and target compounds 1–4.
interest for the potential modulation of biological signals and metabolic activities.[1]
Among the anomeric sugar phosphates, N-acetyl-a-D-glucosamine 1-phosphate
(GlcNAc-1-P) is of particular interest. Besides being the key intermediate in the
biosynthesis of N-linked glycoproteins, it is the metabolic precursor of the bacterial
cell-wall components teichoic acid and mureine. Despite its important biological
implication, only three synthetic analogues of GlcNAc-1-P have been reported. Nicotra
and co-workers synthesised the phosphonate bio-isostere following a multi-step
sequence.[2] The amino function was introduced at the end of the sequence to
overcome the difficulty encountered during the preparation of the corresponding amino
C-glycosyl halides and their subsequent conversion into phosphonate. Junker and
Fessner prepared the diethyl 2-(3’,4’,6’-tri-O-acetyl-2’-deoxy-2’-trifluoroacetamido-a-D-
glucopyranosyl)ethane phosphonate by radical promoted C C bond formation between
diethyl vinyl-phosphonate and the corresponding glycosyl bromide.[3] More recently,
Scha¨fer and Thiem reported the synthesis of an a-C-methylene phosphate analogue of
GlcNAc-1-P by using Kessler’s dianion strategy to prepare the 1-C-a-carboxylic acid
derivative of GlcNAc followed by transformation into the phosphate.[4]
In a previous communication,[5] we reported a concise synthesis of C-ethylene
phosphate 1 and phosphonate 2 of GlcNAc-1-P (Figure 1). In this paper, we describe
the detailed synthesis of compounds 1 and 2 starting from the amino a-C-allyl
glucoside 5, and our investigations into the accessibility of the known C-methylene
phosphate and phosphonate analogues, 3[4] and 4[2] respectively, using the same starting
material. These C-glycosyl phosphates and phosphonates may be considered as
substrate analogues or inhibitors of GlcNAc-1-P uridyltransferase (Glm U)[6] and UDP-
GlcNAc pyrophosphorylase.[7] They may also serve as precursors for the synthesis of
potential inhibitors of N-acetylglycosaminyltransferases.[4]
RESULTS AND DISCUSSION
The stereoselective installation of a C-alkyl phosphate or phosphonate chain at the
anomeric carbon atom of amino sugars with good stereocontrol is not a trivial task.[2]
We decided to use the readily available amino a-C-glycoside 5,[8] exhibiting the
desired stereochemistry at the anomeric center, and to try to convert it into the target
phosphates and phosphonates 1 to 4. As shown in Scheme 1, the C-ethylene analogues
1 and 2 would be obtained via the intermediate alcohol 7. On the other hand, the C-
methylene analogues 3 and 4 can be synthesized from the C-glycoside 6,[9] which is
readily accessible by isomerisation of the double bond in 5.

Synthesis of C-Glycosyl Phosphate and Phosphonate
647
Scheme 1. Retrosynthesis of the target compounds 1–4.
Synthesis of 1 and 2 is shown in Schemes 2 and 3. The amino a-C-allyl glu-
copyranoside 5 was converted into 7 in 85% overall yield through a 2-step process
(oxidative cleavage of the double bond with OsO4 cat/NaIO4, and reduction of the so
obtained aldehyde). The phosphate moiety was introduced satisfactorily either by a one-
step process (treatment of 7 with POCl3) or by a two-step process (phosphorylation with
iPr2N(OBn)2 and subsequent oxidation with mCPBA). Deprotection of 8 and 9 by catalytic
hydrogenolysis led to the desired C-glycopyranosyl phosphate 1 in excellent yield.
Synthesis of the phosphonate analogue 2 was achieved by conversion of alcohol 7
into the bromide 10 upon treatment with CBr4/PPh3 (Scheme 3). The Arbuzov reaction
of 10 with P(OEt)3 afforded 11 (85%) and a small portion of 12, resulting from the
intramolecular substitution of 10. This side reaction was avoided when P(OMe)3 was
used in place of P(OEt)3, thus allowing the use of a lower reflux temperature for the
Arbuzov step. Finally, treatment of 11 and 13 with Me3SiI (20 equiv) in CCl4 led to the
expected phosphonate 2.
To obtain the C-methylene homologues 3 and 4, we intended to use the C-
glycoside 6,[9] obtained by isomerisation of the double bond in 5 (Scheme 4). Thus, the
alkene 6 was oxidatively cleaved using catalytic OsO4 and NaIO4 to provide the
aldehyde 14. However, this a-aldehyde was slowly and irreversibly isomerised to the b-
anomer 15 after work-up. Attempted silica gel purification of 14 led to complete
Scheme 2. i. OsO4, NaIO4, THF/H2O; ii. NaBH4, CH2Cl2/MeOH, 85%; iii. POCl3, THF, 88%; iv.
iPr2NP(OBn)2, tetrazole, THF; v. mCPBA, 97%; vi. Pd/C, MeOH, AcOH cat. quant.

648
Gaurat, Xie, and Vale´ry
Scheme 3. i. CBr4, PPh3, CH2Cl2, RT, quant.; ii. P(OEt)3, reflux; iii. P(OMe)3, reflux, 90%; iv.
TMSI, CCl4, 0°C to RT, quant.
epimerisation, yielding the b-aldehyde 15 in 79% isolated yield. Assignment of the
anomeric configuration of 14 and 15 was based on the observed J1,2 coupling
3
constants (3.1 Hz for 14 and 10.3 Hz for 15), and confirmed by comparison of the d
values for CHO, H-1, H-2 and NH with those in related aldehydes[10,11] (see Table 1).
Indeed, compared to those of the b-isomer, the chemical shifts of the signals for CHO,
H-1, H-2 and NH in the a-isomer are shifted downfield. In addition the aldehyde proton
1
appears as a singlet in the H NMR spectrum of the a-anomer and as a doublet in the
1H NMR spectrum of the b-anomer. Consequently, the crude a-aldehyde 14 was
immediately converted to the alcohol 16[4] in order to avoid epimerisation. Compound
16 was isolated in 80% overall yield from 6 (Scheme 4). Our method may be
considered as an alternative approach to the preparation of 16 in 35% overall yield
from 2-acetamido-3,4,6-tri-O-benzyl-2-deoxy-a-D-glucopyranose.[4] Treatment of 16
with POCl3 failed to furnish the corresponding phosphate. Nevertheless, phosphory-
lation of the hydroxyl group in 16 with iPr2NP(OBn)2 and subsequent oxidation with
Scheme 4. i. OsO4, NaIO4, THF/H2O, quant.; ii. NaBH4, CH2Cl2/MeOH; iii. iPr2NP(OBn)2,
tetrazole, THF; iv. mCPBA, 86%; v. Pd/C, MeOH, AcOH cat. quant.

Synthesis of C-Glycosyl Phosphate and Phosphonate
Table 1. NMR data for selected C-D-glycopyranosyl aldehydes.
649
Compound
Config.
d CHO
H-1
d H-2
d NH (ppm)
14
15
a
b
9.84 (s)
9.45 (d)
9.98 (s)
9.65 (d)
4.03 (J1,2 = 3.1 Hz)
3.56 (J1,2 = 10.3 Hz)
4.40
4.35–4.55
3.90
6.47
5.23
a[10]
b[10]
OBn
O
3.58–3.80
OBn
CHO
CHO
1
BnO
BnO
2
OBn
OBn
O
a[11]
b[11]
9.88 (s)
9.67 (d)
4.32
3.78
OBn
1
2
OBn
mCPBA gave the phosphate 17 in 86% yield. Final hydrogenolysis of the benzyl
protecting groups afforded 3 in quantitative yield.
Compound 4 has been prepared by Nicotra and colleagues through a multi-step
sequence which required introduction of the amino function after installation of a
phosphono group.[2] In order to provide an alternative approach, we tried to convert the
a alcohol 16 into the bromide 19. Unfortunately, all attempts (with PPh3/CBr4 or MsCl/
pyr then LiBr) only resulted in the recovery of the starting material. Indeed, the
bromide 19 formed in the reaction mixture decomposed during workup. Direct
Scheme 5. i. PPh3, CBr4, CH2Cl2; ii. MsCl, Pyr.; iii. LiBr, acetone; iv. P(OMe)3, reflux; v.
NaBH4, CHCl2/MeOH.

650
Gaurat, Xie, and Vale´ry
treatment of mesylate 18 with trimethyl phosphite also failed to afford the desired
phosphonate 20.[12] On the contrary, treatment of the b-alcohol 21 (obtained by
reduction of aldehyde 15 in 89% yield) with PPh3 and CBr4 furnished the expected
bromide 22 in 21% isolated yield (Scheme 5).
In summary, the phosphate and phosphonate analogues of N-acetyl-a-D-glucosamine-
1-phosphate 1 to 3 have been successfully synthesised from the common perbenzylated
amino a-C-allyl glycoside 5. In addition original preparations of a- and b-C-glycosyl
methanol 16 and 21 by oxidation of a-C-methylvinyl glycoside 6, and subsequent
reduction, is reported. These compounds might be useful for the synthesis of potential
inhibitors of glycosyltransferases and more generally of mimics of GlcNAc-1-P.
EXPERIMENTAL
General methods. Melting points were measured with a Thomas-Hoover
1
apparatus. H and 13C NMR spectra were recorded on a Bruker AGH-250 spectrometer
in CDCl3 solution unless noted with tetramethylsilane (Me4Si) as the internal standard.
1
1
Assignments were confirmed by H/1H, H/13C correlations and DEPT 135. Optical
rotations were measured using a Perkin-Elmer 141 polarimeter. Column chromatog-
raphy was performed on E. Merck Silica Gel 60 (230–400 mesh). Analytical thin-layer
chromatography was performed on E. Merck aluminum precoated plates of Silica Gel
60F-254 with detection by UV and by spraying with 6 N H2SO4 and heating about 2
min at 300°C, or by spraying with a solution containing 10% H2SO4 (10 mL), FeCl3
(0.1 g) and 6% orcinol in EtOH (1 mL) and then heating 10 min at 100°C.
Dichloromethane was distilled over CaH2. Tetrahydrofuran was distilled over Na and
benzophenone. Microanalyses were performed at the Service de Microanalyse de
l’Universite´ Pierre et Marie Curie.
4-Acetamido-3,7-anhydro-5,6,8-tri-O-benzyl-2,4-dideoxy-D-glycero-D-ido-octitol
(7). As described,[8] compound 5 was oxidized to the corresponding aldehyde (886
mg, 1.714 mmol) which was dissolved in MeOH (10 mL). NaBH4 (130 mg, 3.428
mmol) was added, and the mixture was stirred for 1 h at rt. The solution was
concentrated, dissolved in EtOAc (30 mL), washed with water, dried over MgSO4,
filtered and concentrated. The residue was purified by column chromatography (1/1
then 2/1 to 1/0 AcOEt/hexane) to afford 7 as a white solid (757 mg, 85%): mp 109–
111°C, Rf 0.23 (AcOEt), [a]D + 15.2 (c 1.0, CH2Cl2); IR (KBr) 3300, 1750, 1650,
1
1525 cmꢀ1. H NMR: d 1.61–1.64 (m, 1H, H-2), 1.73–1.76 (m, 1H, H-2’), 1.81 (s, 3H,
Ac), 2.78 (broad s, 1H, OH), 3.40 (t, 1H, J = 1.5 Hz), 3.53 (dd, 1H, Jgem = 10.3,
J7,8 = 5.8 Hz, H-8), 3.67 (dd, 1H, J = 2.8, J = 1.7 Hz), 3.80 (t, 2H, J = 5.8 Hz, H-1),
4.04 (dd, 1H, Jgem = 10.3, J7,8’ = 8.8 Hz, H-8’), 4.17–4.20 (m, 2H, H-3,4), 4.29 (ddd,
1H, J7,8’ = 8.8, J7,8 = 5.8, J6,7 = 1.0 Hz, H-7), 4.40–4.64 (m, 6H, 3 ꢁ OCH2), 6.91 (d,
1H, J4,NH = 9.8 Hz, NH), 7.18–7.35 (m, 15H, Ph); 13C NMR: d 23.7 (Ac), 32.7 (C-2),
48.5 (C-4), 60.7 (C-1), 67.2 (C-3), 67.4 (C-8), 72.3, 72.6 (CH2), 73.6 (CH), 73.6 (CH2),
74.4 (CH), 75.1 (C-7), 128.0–128.9 (Ph), 137.1, 137.3, 137.6 (Cipso), 170.0 (CO).
Anal. Calcd for C31H37NO6: C, 71.65; H, 7.18; N, 2.70. Found: C, 71.63; H, 7.13;
N, 2.58.

Synthesis of C-Glycosyl Phosphate and Phosphonate
651
(4-Acetamido-3,7-anhydro-5,6,8-tri-O-benzyl-2,4-dideoxy-D-glycero-D-ido-octi-
tol-1-yl) phosphate (8). POCl3 (0.36 mL, 3.850 mmol) was added to a 0°C solution
of 7 (200 mg, 0.385 mmol) in anhydrous THF (12 mL) under argon. After stirring for
20 h at rt, water (2 mL) was added and stirring was continued for 15 min. The
solution was concentrated, the residue was dissolved in CH2Cl2 and washed twice with
1% aq HCl. The organic layer was concentrated and purified by eluting with MeOH
on a resin Dowex H+ (50WX8) column to afford 8 as a white solid (206 mg, 88%):
mp 170–172°C, Rf 0.21 (8.5/0.5/1 CH2Cl2/MeOH/AcOH), [a]D ꢀ 3.5 (c 1.0, CH2Cl2);
1
IR (KBr) 3500, 3300, 1650, 1580 cmꢀ1. H NMR: d 1.70–1.95 (m, 2H, H-2), 1.84 (s,
3H, Ac), 3.54 (s, 1H), 3.65 (s, 1H), 3.71 (dd, 1H, Jgem = 10.0, J7,8 = 6.6 Hz, H-8),
3.89 (dd, 1H, Jgem = 10.0, J7,8’ = 7.0 Hz, H-8’), 4.00–4.30 (m, 5H), 4.38–4.67 (m, 6H,
3 ꢁ OCH2), 6.88 (d, 1H, J4,NH = 9.3 Hz, NH), 7.23–7.35 (m, 15H, Ph), 8.45 (s, 2H,
2 ꢁ OH); 13C NMR: d 22.9 (Ac), 31.7 (C-2), 48.0 (C-4), 63.5 (C-1), 65.0 (C-3), 67.8
(C-8), 72.0, 72.1, 73.3 (CH2), 73.7, 74.1 (C-5,6), 74.7 (C-7), 127.8–128.6 (Ph), 137.4,
137.6, 138.1 (Cipso), 171.5 (CO); 31P NMR (202.46 MHz): d 2.55.
Anal. Calcd for C31H38NO9Pꢂ0.5 H2O: C, 61.18; H, 6.46; N, 2.30. Found: C,
61.31; H, 6.70; N, 2.23.
Di-O-benzyl (4-acetamido-3,7-anhydro-5,6,8-tri-O-benzyl-2,4-dideoxy-D-gly-
cero-D-ido-octitol-1-yl) phosphate (9). Tetrazole (81 mg, 1.155 mmol) and di-O-
benzyl-di-N,N-isopropylphosphorodiamidite (194 mL, 0.578 mmol) were added to a
solution of alcohol 7 (200 mg, 0.385 mmol) in anhydrous THF under argon. After stirring
for 6 h at rt, mCPBA (332 mg, 1.925 mmol) was added, and srirring was continued for
2 h. The solution was diluted with CH2Cl2 (25 mL), washed successively with 10% aq
Na2SO3, 10% aq NaHCO3 and water, dried over MgSO4, and concentrated. Purification
by flash chromatography (1/1 to 1/0 AcOEt/hexane) afforded the title compound as a
white solid (292 mg, 97%): mp 43–44°C, Rf 0.44 (AcOEt), [a]D + 3.3 (c 1.86, CH2Cl2);
IR (KBr) 3300, 1630, 1540 cmꢀ1. 1H NMR: d 1.75–1.90 (m, 2H, H-2), 1.87 (s, 3H, Ac),
3.63 (t, 1H, J = 1.5 Hz), 3.71 (t, 1H, J = 2.0 Hz), 3.75–3.85 (m, 2H, H-8,8’), 4.09–4.26
(m, 5H), 4.40–4.66 (m, 6H, 3 ꢁ OCH2), 5.02 (d, 1H, JC,P = 8.3 Hz, POCH), 5.03 (d, 1H,
JC,P = 8.3 Hz, POCH), 6.69 (d, 1H, J4,NH = 9.8 Hz, NH), 7.25–7.37 (m, 25H, Ph); 31P
NMR (202.46 MHz): d ꢀ 0.15.
Anal. Calcd for C45H50NO9P: C, 69.31; H, 6.46; N, 1.80. Found: C, 69.15; H, 6.66;
N, 1.83.
(4-Acetamido-3,7-anhydro-2,4-dideoxy-D-glycero-D-ido-octitol-1-yl) phosphate
(1). A solution of 8 or 9 (50 mg, 0.082 mmol) in MeOH (3 mL) was hydrogenated at
atmospheric pressure in the presence of 10% palladium on charcoal (10 mg) for 24 h. The
catalyst was filtered off, and the filtrate concentrated to give 29 mg (100%) of the title
compound as a white solid: mp 198–200°C, Rf 0.64 (8/1 iPrOH/AcONH4 1 M), [a]D + 5.2
1
(c 0.7, DMSO); IR (KBr) 3300, 1650, 1580 cmꢀ1. H NMR (D2O): d 1.90 (dtd, 1H,
Jgem = 14.7, J1,2 = 7.7, J2,3 = 3.5 Hz, H-2), 2.07 (s, 3H, Ac), 2.13 (dddd, 1H, Jgem = 14.7,
J1’,2’ = 11.5, J1,2’ = 5.3, J2’,3 = 9.1 Hz, H-2’), 3.46 (t, 1H, J = 9.1 Hz, H-6), 3.63 (ddd, 1H,
J6,7 = 9.1, J7,8 = 5.0, J7,8’ = 2.2 Hz, H-7), 3.76 (t, 1H, J = 9.1 Hz, H-5), 3.77 (dd, 1H,
Jgem = 12.3, J7,8 = 5.0 Hz, H-8), 3.89 (dd, 1H, Jgem = 12.3, J7,8’ = 2.2 Hz, H-8’), 3.95–
4.11 (m, 3H, H-1,1’,4), 4.27 (ddd, 1H, J2’,3 = 9.1, J3,4 = 5.7, J2,3 = 3.5 Hz, H-3); 13C NMR

652
Gaurat, Xie, and Vale´ry
(D2O): d 22.5 (Ac), 26.2 (d, JC,P = 6.6 Hz, C-2), 53.7 (C-4), 61.6 (C-8), 63.0 (d, JC,P = 4.6
Hz, C-1), 71.0, 71.2, 71.3 (C-3,5,6), 73.5 (C-7), 175.1 (CO); 31P NMR (202.46 MHz,
D2O): d 2.82.
Anal. Calcd for C10H20NO9Pꢂ1.5H2O: C, 33.71; H, 6.51; N, 3.93. Found: C, 33.80;
H, 6.14; N, 3.80.
4-Acetamido-3,7-anhydro-5,6,8-tri-O-benzyl-1-bromo-1,2,4-trideoxy-D-glycero-
D-ido-octitol (10). To a solution of alcohol 7 (200 mg, 0.385 mmol) in anhydrous
CH2Cl2, were added PPh3 (202 mg, 0.77 mmol) and CBr4 (281 mg, 0.847 mmol) under
an argon atmosphere. After stirring for 1 h at rt, the reaction mixture was diluted with
CH2Cl2 (15 mL) and washed twice with water. The organic layer was then dried
(MgSO4) and concentrated. Purification by column chromatography (1/1 AcOEt/
hexane) afforded 10 as a white solid (224 mg, 100%): mp 101–102°C, Rf 0.61 (1/1
cyclohexane/AcOEt), [a]D + 13.9 (c 1.0, CH2Cl2); IR (KBr) 3300, 1750, 1650, 1525
1
cmꢀ1. H NMR: d 1.78–2.05 (m, 2H, H-2), 1.79 (s, 3H, Ac), 3.49 (dd, 1H, J1,2 = 7.6,
J1,2’ =5.8 Hz, H-1), 3.57 (t, 1H, J = 1.3 Hz, H-6), 3.64 (ddd, 1H, J4,5 = 4.0, J5,6 = 1.3,
J5,7 = 2.8 Hz, H-5), 3.77 (dd, 1H, Jgem = 10.0, J7,8 = 7.0 Hz, H-8), 3.90 (dd, 1H,
Jgem = 10.0, J7’,8 = 7.1 Hz, H-8’), 4.16–4.28 (m, 3H, H-3,4,7), 4.39–4.67 (m, 6H,
3 ꢁ OCH2), 6.64 (d, 1H, J4,NH = 9.5 Hz, NH), 7.22–7.37 (m, 15H, Ph); 13C NMR:
d 23.4 (Ac), 30.2 (C-2), 34.3 (C-1), 47.3 (C-4), 65.9 (C-3), 67.9 (C-8), 71.9, 72.1, 73.3
(CH2), 73.5 (C-6), 74.2 (C-5), 75.2 (C-7), 127.6–129.7 (Ph), 137.3, 137.5, 138.1
(Cipso), 169.9 (CO).
Anal. Calcd for C31H36BrNO5: C, 63.92; H, 6.23; N, 2.40. Found: C, 63.56; H,
6.21; N, 2.41.
Di-O-ethyl (4-acetamido-3,7-anhydro-5,6,8-tri-O-benzyl-1,2,4-trideoxy-D-gly-
cero-D-ido-octitol-1-yl) phosphonate (11). A mixture of 10 (220 mg, 0.378 mmol)
and triethyl phosphite (10 mL) was heated under reflux for 15 h, then concentrated in
vacuo. Purification by column chromatography (0/1 to 1/9 acetone/AcOEt) afforded
205 mg (85%) of 11 (white solid) and 19 mg (10%) of pyrrolidine 12. Compound 11:
mp 90–92°C, Rf 0.61 (1/1 AcOEt/hexane), [a]D + 7.0 (c 1.0, CH2Cl2); IR (KBr) 3330,
1
1650, 1525 cmꢀ1. H NMR: d 1.23 (t, 6H, J = 7.0 Hz, 2 ꢁ CH3), 1.30–1.85 (m, 4H,
H-1,2), 1.78 (s, 3H, Ac), 3.53 (t, 1H, J = 1.5 Hz), 3.63 (m, 1H), 3.68 (dd, 1H,
Jgem = 15.6, J7,8 = 8.7 Hz, H-8), 3.79 (dd, 1H, Jgem = 15.6, J7’,8 = 6.9 Hz, H-8’), 3.81–
3.86 (m, 1H, H-3), 3.99 (dt, 2H, J = 7.0, JH,P = 14.8 Hz, CH2-OP), 4.00 (dt, 2H,
J = 7.0, JH,P = 15.1 Hz, CH2-OP), 4.08–4.19 (m, 2H, H-4,7), 4.37–4.58 (m, 6H,
3 ꢁ OCH2), 6.45 (d, 1H, J4,NH = 9.6 Hz, NH), 7.23–7.36 (m, 15H, Ph); 13C NMR:
d 16.3 (d, JC,P = 5.6 Hz, CH3), 21.4 (d, JC,P = 142.5 Hz, C-1), 23.1 (Ac), 23.9 (C-2),
47.5 (C-4), 61.4 (d, JC,P = 6.1 Hz, CH2-OP), 67.6 (C-8), 68.0 (d, JC,P = 17.4 Hz, C-3),
71.8, 72.1, 73.1 (CH2), 73.3, 74.4, 74.6 (CH), 127.5–128.4 (Ph), 137.2, 137.4, 137.9
(Cipso), 169.7 (CO); 31P NMR (202.46 MHz): d 33.02.
Anal. Calcd for C35H46NO8P: C, 65.71; H, 7.25; N, 2.19. Found: C, 65.31; H, 7.12;
N, 2.08.
Di-O-methyl (4-acetamido-3,7-anhydro-5,6,8-tri-O-benzyl-1,2,4-trideoxy-D-gly-
cero-D-ido-octitol-1-yl) phosphonate (13). Compound 13 was prepared from 10
and trimethyl phosphite as described for 12. Yield 90%, mp 51–52°C, Rf 0.12

Synthesis of C-Glycosyl Phosphate and Phosphonate
653
1
(AcOEt), [a]D + 9.7 (c 0.95, CH2Cl2). H NMR: d 1.61–1.90 (m, 4H, H-1,2), 1.82 (s,
3H, Ac), 3.56 (s, 1H), 3.65 (m, 1H), 3.70 (dd, 1H, Jgem = 10.0, J7,8 = 6.8 Hz, H-8),
3.75 (d, 6H, JH,P = 10.8 Hz, 2 ꢁ CH3-OP), 3.82 (dd, 1H, Jgem = 10.0, J7’,8 = 7.0 Hz,
H-8’), 3.90 (m, 1H, H-3), 4.10–4.22 (m, 2H, H-4,7), 4.41–4.63 (m, 6H, 3 ꢁ OCH2),
6.48 (d, 1H, J4,NH = 9.5 Hz, NH), 7.21–7.35 (m, 15H, Ph); 13C NMR: d 21.0 (d,
JC,P = 142.3 Hz, C-1), 23.6 (Ac), 24.3 (d, JC,P = 4.0 Hz, C-2), 48.0 (C-4), 68.1 (C-8),
68.6 (d, JC,P = 17.3 Hz, C-3), 72.4, 72.6, 73.6 (CH2), 73.6, 75.0 (CH), 127.5–128.4
(Ph), 137.2, 137.5, 137.9 (Cipso), 169.7 (CO); 31P NMR (202.46 MHz): d 35.9.
Anal. Calcd for C33H42NO8P: C, 64.80; H, 6.92; N, 2.29. Found: C, 64.43; H, 7.13;
N, 2.09.
(4-Acetamido-3,7-anhydro-1,2,4-trideoxy-D-glycero-D-ido-octitol-1-yl) phospho-
nate (2). To a solution of 11 (180 mg, 0.281 mmol) in anhydrous CCl4 (7 mL) at 0°C
under an argon atmosphere, was added TMSI (0.801 mL, 5.62 mmol). After stirring for
1 h at rt, water (5 mL) was added and stirring continued during 30 min. The mixture
was then separated. The aqueous layer was washed twice with Et2O and concentrated
to afford 11 as a solid which was recrystallized from THF (88 mg, 100%): mp 132–
1
134°C (THF), Rf 0.79 (8/1 iPrOH/AcONH4 1 M), [a]D + 55.8 (c 0.87, H2O). H NMR
(D2O): d 1.50–1.80 (m, 2H, H-1), 1.65–1.90 (m, 2H, H-2), 1.92 (s, 3H, Ac), 3.38 (t,
1H, J = 9.0 Hz, H-6), 3.48 (ddd, 1H, J6,7 = 9.0, J7,8 = 5.0, J7,8’ = 1.8 Hz, H-7), 3.68
(dd, 1H, Jgem = 12.0, J7,8 = 5.0 Hz, H-8), 3.71 (t, 1H, J = 9.0 Hz, H-5), 3.84 (dd, 1H,
Jgem = 12.0, J7,8’ = 1.8 Hz, H-8’), 3.95 (dd, 1H, J3,4 = 5.6, J4,5 = 9.0 Hz, H-4), 3.95–
4.08 (m, 1H, H-3); 13C NMR (D2O): d 20.6 (C-2), 23.2 (Ac), 24.9 (d, JC,P = 134.5, C-
1), 54.6 (C-4), 62.3 (C-8), 71.8, 72.2 (C-5,6), 73.6 (C-7), 75.6 (C-3), 175.1 (CO); 31P
NMR (202.46 MHz, D2O): d 32.04.
Anal. Calcd for C10H20NO8P: C, 38.34; H, 6.44; N, 4.47. Found: C, 38.60; H, 6.66;
N, 4.31.
3-Acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-aldehydo-D-glycero-D-ido-
heptopyranose (14) and 3-acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-alde-
hydo-D-glycero-D-gulo-heptopyranose (15). To a solution of alkene 6[9] (255 mg,
0.495 mmol) in a mixture of THF/H2O (2/1, 2 mL), were added OsO4 (4% solution in
t-BuOH, 124 mL) and NaIO4 (529 mg, 2.475 mmol). After stirring for 20 h at rt, the
mixture was concentrated, diluted in CH2Cl2, washed with water, 5% aq Na2S2O3 and
brine, dried over MgSO4, filtered and concentrated to give the aldehyde 14 as an oil
(244 mg, 98%). Purification by column chromatography (AcOEt) epimerised the a-
aldehyde 14 to the b-aldehyde 15 which was isolated as an oil (197 mg, 79%).
1
Compound 14: Rf 0.50 (AcOEt). H NMR: d 1.75 (s, 3H, Ac), 3.50–3.75 (m,
4H), 4.11 (td, 1H, J = 3.4, J = 6.0 Hz, H-6), 4.03 (d, 1H, J2,3 = 3.1 Hz, H-2), 4.35–
4.55 (m, 7H, H-3, 3 ꢁ OCH2), 6.47 (d, 1H, J3,NH = 9.5 Hz, NH), 7.19–7.36 (m,
15H, Ph), 9.84 (s, 1H, H-1); 13C NMR: d 23.0 (Ac), 46.6 (C-3), 67.1 (C-7), 72.6,
72.8, 73.2 (CH2), 73.9, 75.1, 75.4 (CH), 127.7–128.5 (Ph), 137.1, 137.3, 137.7
(Cipso), 170.0, 199.5 (CO).
1
Compound 15: Rf 0.50 (AcOEt). H NMR: d 1.68 (s, 3H, Ac), 3.43–3.48 (m, 1H),
3.56 (dd, 1H, J1,2 = 2.5, J2,3 = 10.3 Hz, H-2), 3.60–3.67 (m, 4H), 3.90 (ddd, 1H,
J2,3 = 10.3 Hz, J3,NH = 8.3, J3,4 = 8.1 Hz, H-3), 4.45–4.60 (m, 4H, 2 ꢁ OCH2), 4.72
(d, 1H, Jgem = 9.6 Hz, CH-O), 4.80 (d, 1H, Jgem = 9.6 Hz, CH-O), 5.23 (d, 1H,

654
Gaurat, Xie, and Vale´ry
J3,NH = 8.3 Hz, NH), 7.10–7.30 (m, 15H, Ph), 9.45 (d, 1H, J1,2 = 2.5 Hz, H-1); 13C
NMR: d 22.1 (Ac), 50.0 (C-3), 67.5 (C-7), 72.5, 73.5, 73.9 (CH2), 78.6, 79.3, 82.3, 82.4
(CH), 128.2–129.1 (Ph), 136.6, 136.9 (Cipso), 170.6, 197.5 (CO).
Anal. Calcd for C30H33NO6: C, 71.55; H, 6.60; N, 2.78. Found: C, 71.39; H, 6.71;
N, 2.88.
3-Acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-D-glycero-D-ido-heptitol
(16).[4] NaBH4 (30 mg, 0.789 mmol) was added to a solution of 14 (206 mg, 4 mmol)
in a mixture of MeOH/CH2Cl2 (1/1, 2 mL), and the mixture was stirred for 20 h at rt.
The solution was concentrated, dissolved in AcOEt, washed with water, dried (MgSO4),
filtered and concentrated. Purification by column chromatography (2/1 to 1/0 AcOEt/
cyclohexane) afforded 16 (165 mg, 80%) as a white solid: mp 90°C, Rf 0.55 (AcOEt),
[a]D + 20.7 (c 1.1, CH2Cl2), + 19.2 (c 1.0, acetone), Lit.[4] mp 89–91°C, [a]D + 20.1 (c
1.22, acetone); IR (KBr) 3421, 3324, 3107, 3059, 3035, 1685, 1660 cmꢀ1
.
Di-O-benzyl (3-acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-D-glycero-D-
ido-heptitol-1-yl) phosphate (17). Compound 17 was prepared from 16 as described
for 9. Purification by flash chromatography (1/1 to 1/0 AcOEt/hexane) afforded the
title compound as an oil (86%): Rf 0.66 (AcOEt), [a]D + 0.0, [a]Hg
ꢀ 0.44,
[a]Hg
ꢀ 11.3 (c 1.13, CH2Cl2); IR (KBr) 3300, 1650, 1525 cmꢀ1(546)
.
1H NMR:
(365)
d 1.77 (s, 3H, Ac), 3.60 (m, 1H), 3.66 (t, 1H, J = 3.1 Hz), 3.72–3.76 (m, 2H, H-
8,8’), 4.01–4.07 (m, 2H, H-1), 4.17 (ddd, 1H, J = 7.4, J = 5.3, J = 2.2 Hz, H-2), 4.22–
4.27 (m, 2H, H-3,6), 4.40–4.61 (m, 6H, 3 ꢁ OCH2), 4.95–5.08 (m, 4H, 2 ꢁ POCH2),
6.54 (d, 1H, J = 9.5 Hz, NH), 7.19–7.34 (m, 25H, Ph); 13C NMR: d 23.3 (Ac), 46.0
(C-3), 67.7 (C-7), 67.8 (d, JC,P = 7.0 Hz, C-2), 67.9 (d, JC,P = 4.0 Hz, C-1), 69.3 (d,
JC,P = 5.0 Hz, CH2OP), 72.5, 72.7, 73.7 (CH2), 73.8, 74.8 (C-4,5), 75.5 (C-6), 127.7–
128.6 (Ph), 136.0 (d, JC,P = 4.5 Hz, Cipso Ph-OP), 137.4, 137.5, 138.1 (Cipso), 169.9
(CO); 31P NMR (202.46 MHz): d 0.01.
Anal. Calcd for C44H48NO9P: C, 69.01; H, 6.32; N, 1.83. Found: C, 68.64; H, 6.41;
N, 1.72.
(3-Acetamido-2,6-anhydro-3-deoxy-D-glycero-D-ido-heptitol-1-yl) phosphate
(3).[4] Compound 17 was deprotected as described for 9 to afford 3 as a white solid
(100%): mp 120–124°C, [a]D + 31.7 (c 0.7, H2O), Litt.[4] [a]D + 29 (c 0.5, H2O). 31P
NMR (202.46 MHz, D2O): d 2.72.
3-Acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-1-O-methylsulfonyl-D-gly-
cero-D-ido-heptitol (18). To a solution of 16 (70 mg, 0.139 mol) in dry pyridine,
was added methanesulfonyl chloride (22 mL, 0.278 mmol) dropwise. After 1 h,
CH2Cl2 was added and the mixture was washed twice with 5% aq HCl and once
with water, dried (MgSO4), and concentrated. Purification by flash chromatography
(AcOEt) afforded the title compound as a white solid (92%): mp 57–58°C, Rf 0.60
1
(AcOEt), [a]D + 9.4 (c 0.85, CH2Cl2); IR (KBr) 3300, 1650, 1525 cmꢀ1. H NMR:
d 1.85 (s, 3H, Ac), 3.01 (s, 3H, Ms), 3.58–3.59 (m, 1H), 3.68 (dd, 1H, Jgem = 10.0,
J6,7 = 6.5 Hz, H-7), 3.63–3.67 (m, 1H), 3.92 (dd, 1H, Jgem = 10.0, J6,7’ = 7.5 Hz,
H-7’), 4.20–4.25 (m, 3H, H-1,2), 4.30–4.40 (m, 2H, H-3,6), 4.40–4.65 (m, 6H,
3 ꢁ OCH2), 6.70 (d, 1H, J3,NH = 9.0 Hz, NH), 7.20–7.40 (m, 15H, Ph); 13C NMR:

Synthesis of C-Glycosyl Phosphate and Phosphonate
655
d 23.7 (Ac), 38.1 (Ms), 46.1 (C-3), 67.2 (C-2), 67.7 (C-7), 70.9 (C-1), 72.4, 72.6
(CH2), 73.3 (CH), 73.8 (CH2), 74.1 (CH), 75.7 (C-6), 127.6–128.7 (Ph), 137.0,
137.2, 137.9 (Cipso), 169.9 (CO).
Anal. Calcd for C31H37NO8S: C, 63.79; H, 6.39; N, 2.40. Found: C, 63.73; H, 6.61;
N, 2.46.
3-Acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-D-glycero-D-gulo-heptitol
(21). Compound 21 was prepared from 15 as described for 16. Yield 89%, mp 142–
1
143°C, Rf 0.19 (AcOEt), [a]D + 23.1 (c 1.16, CH2Cl2). H NMR: d 1.74 (s, 3H, Ac),
3.11 (dd, 1H, J3,4 = 10.0, J4,5 = 2.5 Hz, H-4), 3.43–3.71 (m, 7H), 3.86 (td, 1H,
J2,3 = J3,4 = 10.0, J3,NH = 8.0 Hz, H-3), 4.52–4.90 (m, 6H, 3 ꢁ OCH2), 4.94 (d, 1H,
J3,NH = 8.0 Hz, NH), 7.19–7.42 (m, 15H, Ph); 13C NMR: d 23.2 (Ac), 50.9 (C-3), 61.9,
69.2 (C-1,7), 73.6, 74.2, 75.1 (CH2), 79.1, 79.2 (CH), 79.9 (C-4), 82.0 (CH), 127.8–
128.9 (Ph), 137.8, 138.0, 138.4 (Cipso), 171.8 (CO).
Anal. Calcd for C30H35NO6: C, 71.27; H, 6.98; N, 2.77. Found: C, 71.55; H, 7.01;
N, 2.68.
3-Acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-1-bromo-4,5,7-tri-O-benzyl-1,3-
dideoxy-D-glycero-D-gulo-heptitol (22). Compound 22 was prepared from 21 as de-
scribed for 10. Purification by flash chromatography (1/1 AcOEt/cyclohexane) afforded
the title compound as a white solid (21%): mp 177–178°C, Rf 0.76 (AcOEt), [a]D + 16.0
(c 1.0, CH2Cl2). 1H NMR: d 1.74 (s, 3H, Ac), 3.32–3.47 (m, 3H, CH + H-1), 3.52–3.71
(m, 6H), 4.50–4.82 (m, 6H, 3 ꢁ OCH2), 5.02 (d, 1H, J3,NH = 7.0 Hz, NH), 7.13–7.32
(m, 15H, Ph); 13C NMR: d 23.6 (Ac), 33.4 (C-1), 54.7 (C-3), 68.8 (C-7), 73.6, 74.7, 75.0
(CH2), 78.2, 78.9, 79.4, 82.2 (CH), 127.7–128.8 (Ph), 138.1, 138.4, 138.5 (Cipso),
170.8 (CO).
Anal. Calcd for C30H34BrNO6: C, 61.86; H, 5.53; N, 2.40. Found: C, 61.58; H,
5.66; N, 2.60.
REFERENCES
1. Nicotra, F. Synthesis of glycosyl phosphate mimics. In Carbohydrate Mimics:
Concepts and Methods; Chapleur, Y., Ed.; Wiley-VCH: Weinheim, 1998; 67–85.
2. Casero, F.; Cipolla, L.; Lay, L.; Nicotra, F.; Panza, L.; Russo, G. Stereoselective
synthesis of the isosteric phosphono analogues of N-acetyl-a-D-glucosamine 1-
phosphate and N-acetyl-a-D-mannosamine 1-phosphate. J. Org. Chem. 1996, 61
(10), 3428–3432.
3. Junker, H.D.; Fessner, W.D. Diastereoselective synthesis of C-glycosylphos-
phonates via free-radical glycosylation. Tetrahedron Lett. 1998, 39 (3/4), 269–272.
4. Scha¨fer, A.; Thiem, J. Synthesis of novel donor mimetics of UDP-Gal, UDP-
GlcNAc, and UDP-GalNAc as potential transferases inhibitors. J. Org. Chem. 2000,
65 (1), 24–29.
5. Gaurat, O.; Xie, J.; Vale´ry, J.M. A concise synthesis of C-glycosyl phosphate and
phosphonate analogues of N-acetyl-a-D-glucosamine 1-phosphate. Tetrahedron
Lett. 2000, 41 (8), 1187–1189.
6. Brown, K.; Pompeo, F.; Dixon, S.; Mengin-Lecreulx, D.; Cambillau, C.; Bourne, Y.

656
Gaurat, Xie, and Vale´ry
Crystal structure of the bifunctional N-acetylglucosamine 1-phosphate uridyltrans-
ferase from Escherichia coli: a paradigm for the related pyrophosphorylase
superfamily. EMBO J. 1999, 18, 4096–4107.
7. Szumilo, T.; Zeng, Y.; Pastuszak, I.; Drake, R.; Szumilo, H.; Elbein, A. Purification
to homogeneity and properties of UDP-GlcNAc (GalNAc) pyrophosphorylase. J.
Biol. Chem. 1996, 271, 13147–13154.
8. Grugier, J.; Xie, J.; Duarte, I.; Vale´ry, J.M. Synthesis of 2-(N-acetylamino)-2-
deoxy-C-glucopyranosyl nucleosides as potential inhibitors of chitin synthases. J.
Org. Chem. 2000, 65 (4), 979–984.
9. Xie, J. Synthesis of new sugar aminoacid derivatives of D-glucosamine. Carbohydr.
Res. 2003, 338, 399–406.
10. Dondoni, A.; Scherrmann, M.C. Thiazole-based synthesis of formyl C-glycosides.
J. Org. Chem. 1994, 59 (20), 6404–6412.
11. Kobertz, W.R.; Bertozzi, C.R.; Bednarski, M.D. An efficient method for the
synthesis of a- and b-C-glycosyl aldehydes. Tetrahedron Lett. 1992, 33 (6), 737–
740.
12. Frische, K.; Schmidt, R.R. Glycal-1-ylmethylphosphonates-precursors of glycosyl-
transferase inhibitors. Liebigs Ann. Chem. 1994, 297–303.
Received February 24, 2003
Accepted July 24, 2003