Synthesis of methyl β-D-arabinofuranoside 5-[1D (and L)-myo-inositol 1-phosphate], the capping motif of the lipoarabinomannan of Mycobacterium smegmatis

Article abstract of DOI:10.1016/S0008-6215(99)00078-6

The total synthesis of methyl β-D-arabinofuranoside 5-(myo-inositol 1-phosphate), the capping motif of the lipoarabinomannan (LAM) of Mycobacterium smegmatis, has been completed. The stereoselective synthesis of β-D-arabinofuranosides has been achieved vi

Full text of DOI:10.1016/S0008-6215(99)00078-6

www.elsevier.nl/locate/carres
Carbohydrate Research 317 (1999) 110–118
Synthesis of methyl b-
D
-arabinofuranoside 5-[1
D
(and
L
)-myo-inositol 1-phosphate], the capping motif of the
lipoarabinomannan of Mycobacterium smegmatis
Je´roˆme De´sire´, Jacques Prandi *
Institut de Pharmacologie et de Biologie Structurale, Centre National de la Recherche Scientifique,
205 route de Narbonne, F-31077 Toulouse, France
Received 9 November 1998; accepted 24 March 1999
Abstract
The total synthesis of methyl b-
lipoarabinomannan (LAM) of Mycobacterium smegmatis, has been completed. The stereoselective synthesis of
b- -arabinofuranosides has been achieved via an internal aglycon delivery approach using Ogawa and Ito’s method.
D-arabinofuranoside 5-(myo-inositol 1-phosphate), the capping motif of the
D
Coupling with enantiomeric myo-inositol derivatives gave the diastereoisomeric title compounds in good overall
yield. Comparison with the natural product firmly established the proposed structure for the capping of the LAM but
left the absolute configuration of the myo-inosityl moiety undetermined. © 1999 Elsevier Science Ltd. All rights
reserved.
Keywords: Mycobacterium smegmatis; Lipoarabinomannan; b-
D
-Arabinofuranoside; Sugar phosphates
isolated from various species of mycobacteria
were observed and these variations have been
tentatively correlated to small structural dif-
ferences. In the LAM isolated from the non-
pathogenic species Mycobacterium smegmatis
[5], the hydroxymethyl group of the terminal
1. Introduction
Lipoarabinomannans (LAMs) are highly
antigenic oligosaccharide components, ubiqui-
tous to the Mycobacterium genus, which play
a key role in the immunopathogenicity of
these microorganisms [1]. They are isolated
from the cell wall and share common struc-
tural features: a phosphatidyl-myo-inositol an-
chor and an oligomannan core substituted by
an arabinan domain [1]. The nonreducing
ends of this arabinan moiety are terminated
b- -arabinofuranosides is esterified by a phos-
D
phate group carrying a 1-myo-inositol of un-
determined series. This LAM was found to be
an activator of TNF-a production by human
macrophages (THP-1 cell line) at a concentra-
tion of 10 mg/mL and this activity was reduced
after a mild basic treatment [5]. On the other
hand, LAMs isolated from M. tuberculosis
and M. bo6is BCG were devoid of this activ-
ity; in these compounds, the caps on the ter-
minal arabinofuranosides are small a-(1“2)-
linked oligomannosides [2,3].
by b- -arabinofuranosidic units substituted at
D
C-5 by small motifs, highly variable on the
mycobacterial species [2–5]. Strong modula-
tions of the biological properties of the LAMs
We now report the synthesis of methyl b-
D
-
* Corresponding author. Fax: +33-5-6117-5994.
E-mail address: prandi@ipbs.fr (J. Prandi)
arabinofuranoside 5-[1 (and )-myo-inositol
D
L
0008-6215/99/$ - see front matter © 1999 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(99)00078-6

J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
111
1-phosphate], 1 and 2, the capping motif of
the lipoarabinomanan of M. smegmatis. We
also report the use of the internal aglycon
delivery approach described by Ogawa et al.
[6,7] for the first stereoselective synthesis of
ous benzylation procedures (pMeOBnCl,
NaH, DMF or pMeOBnCl, Bu4NHSO4,
CH2Cl2–NaOH [23]) were ineffective. This hy-
droxyl group was eventually benzylated in 59%
yield with p-methoxybenzyl bromide [24] and 2
-tert-butylimino-2-diethylamino-1,3-dimethyl-
b- -arabinofuranosides [8], compounds of in-
D
terest as potential arabinosyl transferase in-
hibitors [9–11].
perhydro-1,3,2-diazaphosphorine
(BEMP)
[25–27] in acetonitrile. Compound 6 was first
treated with 1.5 equivalent of 2,3-dichloro-5,6-
dicyanobenzoquinone (DDQ) and 1 equivalent
of methanol and the very sensitive intermediate
mixed acetal [6,7] was then activated with
iodonium dicollidine perchlorate (IDCP) [28] in
the presence of a catalytic amount of
trimethylsilyltriflate (Me3SiOTf) to promote
the glycosylation [29,30]. Under these condi-
tions, the TIPS protecting group was lost and
only degradation products were observed. The
TIPS group in 6 was thus removed and replaced
by acetyl groups through successive treatment
2. Results and discussion
Synthesis of i- -arabinofuranosides.—
D
Among the numerous methods of intramolec-
ular glycosylation recently described for the
stereoselective synthesis of b-mannosides [12–
16], Ogawa’s procedure [6,7], which uses the
p-methoxybenzylidene group as temporary
linker, was found to be the most convenient
[17,18] (Scheme 1).
of
6
with tetrabutylammonium fluoride
Treatment of methyl tri-O-benzoyl-a- -ara-
D
(nBu4NF) in tetrahydrofuran (THF) and acety-
lation with acetic anhydride in pyridine to 8
(77%, two steps). Compound 8 was then se-
quentially treated as above with DDQ–
methanol and IDCP–Me3SiOTf to give
binofuranoside [19] with ethanethiol and boron
trifluoride–etherate gave the ethyl thiogly-
coside 3 in 80% yield. The benzoyl protecting
groups were removed with a catalytic amount
of sodium methoxide and simultaneous protec-
tion of the 3 and 5 positions of triol 4 was
achieved with 1 equivalent of 1,3-dichloro-
1,1,3,3-tetraisopropyldisiloxane (TIPS chlo-
ride) in pyridine to give 5 in 75% yield [20–22].
Introduction of the p-methoxybenzyl ether at
C-2 of 5 was somewhat troublesome and vari-
anomerically pure methyl 3,5-di-O-acetyl-b-
D
-
arabinofuranoside (9) in 51% yield from 8. The
b-( ) configuration of 9 was established from
D
1
the H and 13C NMR data (lH-1 4.90 ppm,
JH-1,H-2 4.5 Hz, lC-1 102.3 ppm) [31–34];
deacetylation then furnished methyl b-
D-ara-
binofuranoside (10) in 80% yield [35].
In the same manner, the b-linked diarabino-
furanoside 14 was obtained in two steps from
8. Reaction of 8 with 1.5 equivalents of DDQ
and 1.05 equivalents of methyl 2,3-di-O-acetyl-
a- -arabinofuranoside [36] gave the intermedi-
D
ate acetal in 71% yield after purification on
silica, intramolecular glycosylation as above
gave disaccharide 14 with the b-( ) configura-
D
tion at the new anomeric center as the only
isolated coupling product (74%, lH-1% 5.12 ppm,
JH-1%,H-2%, 4.5 Hz, lC-1% 101.6 ppm) [31–34].
Synthesis of 5-(myo-inositol 1-phosphate) i-
D-arabinofuranosides.—The primary hydroxyl
group of methyl b-
D
-arabinofuranoside (10)
was selectively silylated with 1 equivalent of
tert-butyldiphenylsilyl chloride in dimethyl-
formamide (65%) and the 2 and 3 positions of
Scheme 1.

112
J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
Scheme 2.
11 were then benzylated (BnBr, NaH) to give
12 in 87% yield. Deprotection of the tert-
butyldiphenylsilyl group with nBu4NF in THF
afforded the 2,3-diprotected arabinofuranoside
acceptor 13 in 74% yield.
hydrogenolysis of the benzyl groups gave the
phosphodiester 2.
The 1H, 13C and 31P NMR spectra for the two
diastereoisomers 1 and 2 were then recorded
and compared with the reported values for the
LAM of M. smegmatis [5]. NMR data for 1 and
2 were found to be very similar and very close
to the data for the natural product (see Table
3), only minor differences could be observed.
Enantiomeric 1
L
- and 1 -1,2,4,5,6-penta-O-
D
benzyl-myo-inositol (15 and 17) were obtained
from the racemic compound by resolution with
(1S)-camphanic acid chloride [37,38], and each
enantiomer was then carried out along the
1
Furthermore, in the H NMR spectrum of the
synthesis. The 1
L
-1,2,4,5,6-penta-O-benzyl-
natural product, a severe crowding in the region
of the sugar protons was observed, which
precluded complete assignment of the signals of
the arabinofuranoside moiety. Although these
data unambiguously confirmed the proposed
structure for the capping motif of the LAM of
M. smegmatis, no correlation could be drawn
with the absolute configuration of the myo-
inositol moiety in the natural product, which
then remains undetermined.
myo-inositol (15) {[h]2D5 −9.8°, (c 1.08, CHCl3),
lit. −9.0° [37]} was converted to the H-phos-
phonate 16 by treatment with 3 equivalents of
phosphorus trichloride and imidazole [39–41]
in 57% yield {[h]2D5 +22° (c 1.49, CHCl3)}.
Activation of the H-phosphonate 16 with 3
equivalents of pivaloyl chloride in pyridine and
,
coupling with 13 in the presence of 4 A molec-
ular sieves gave the phosphodiester 19 after
oxidation with iodine (67% yield, two steps).
Final deprotection of the benzyl groups was
achieved by hydrogenolysis with palladium hy-
droxide on carbon and gave compound 1 in
79% yield (Scheme 2).
A similar sequence was carried out from 17
{[h]2D5 +9.7° (c 1.00, CHCl3), lit.+9.2° [37]}.
Conversion to the H-phosphonate 18 {56%,
[h]2D5 −23° (c 1.65, CHCl3)}, coupling with 13
under pivaloyl chloride activation followed by
iodine oxidation to 20 (69%, two steps) and
No TNF-a production by macrophages (cell
line THP-1) could be detected with compounds
1 and 2 up to a concentration of 100 mg/mL.
Finally, compounds 1 and 2 were shown to be
stable to the basic conditions used on the
natural product (NaOH, 0.1 N, 40 °C, 2 h).
This result indicates that the observed diminu-
tion of TNF-a production by macrophages
after basic treatment of the natural LAM was
not caused by some hydrolysis of the inositol
phosphate [5].

J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
113
3. Conclusions
imeter. Mass spectra were obtained on a
Perkin–Elmer SCIEX API 300 (ion spray, IS)
or Finnigan MAT TSQ 700 (electrospray,
negative mode, ES). Elemental analyses were
carried out at the Service Central de Micro-
analyse du CNRS at Vernaison or at the
‘Laboratoire de Chimie de Coordination du
CNRS’ at Toulouse.
The internal aglycon delivery approach de-
veloped by Ogawa et al. [6,7] was found to
offer a general and stereospecific access to
b-
D
-arabinofuranoside compounds. This was
used for the synthesis of methyl b-
D-arabino-
furanoside 5-(myo-inositol 1-phosphate), the
proposed structure for the caps of the LAMs
isolated from M. smegmatis which was fully
confirmed. In order to ascertain the absolute
configuration of the myo-inositol in these
caps, derivatization of compounds 1 and 2 for
comparison with fragments obtained from the
degradation of the LAM of M. smegmatis are
currently under progress and will be reported
in the future.
Ethyl
binofuranoside (3).—Methyl 2,3,5-tri-O-ben-
zoyl-a- -arabinofuranoside [19] (12.0 g, 25
2,3,5-tri-O-benzoyl-1-thio-h-
D-ara-
D
mmol) in anhyd CH2Cl2, EtSH (2.80 mL, 38
mmol) and BF3·Et2O (3.83 mL, 30 mmol)
were kept for 16 h at room temperature (rt),
then neutralized with triethylamine and con-
centrated. Chromatography (4:1 petroleum
ether–EtOAc) gave 3 as a colorless oil (10.35
1
g, 81%); [h]2D5 +22.5° (c 1.29, CHCl3); H
13
NMR data see Table 1; C NMR (CDCl3): l
166.05, 165.50, 165.33 (CꢀO), 133.50, 133.44,
132.97, 129.93, 129.87, 129.75, 129.62, 129.56,
128.85, 128.40, 128.21, 127.80 (Ar), 88.05 (C-
1), 82.90, 80.60, 78.03 (C-2, C-3, C-4), 63.35
(C-5), 25.21 (CH2S), 14.80 (CH3CH2S). Anal.
Calcd for C28H26O7S: C, 66.39; H, 5.17.
Found: C, 66.21; H, 5.28.
4. Experimental
General methods.—Reactions were per-
formed under argon in anhyd purified sol-
vents. NMR spectra were recorded on Bruker
DPX 250, AMX 250 (operating at 250 MHz
1
for H and at 62.9 MHz for 13C) and DMX
1
Ethyl 3,5-O-(1,1,3,3-tetraisopropyldisilox-
500 (operating at 500 MHz for H, at 125.8
MHz for 13C, and at 202.46 MHz for 31P)
spectrometers, chemical shifts are expressed in
ane-1,3-diyl)-1-thio-h- -arabinofuranoside
D
(5).—Compound 3 (8.0 g, 16 mmol) was
treated at rt with MeONa in MeOH (0.1 M,
30 mL). After completion of the reaction, the
mixture was neutralized with Amberlyst IR
ppm, from Me4Si for H and 13C spectra and
1
31
from H3PO4 for P spectra. Optical rotations
were recorded on a Perkin–Elmer 41 polar-
Table 1
1H NMR chemical shifts (l), apparent multiplicities and coupling constants in Hz for compounds 3, 5, 6 and 8
H
1
3
5
6
8
5.64 d
5.09 d
J1,2 5.5
4.10 dd
J2,3 6.5
4.20 dd
J3,4 8.2
3.89 ddd
J 3.0, 6.2
3.97 m
5.21 d
5.37 d
J1,2 2.2
3.94 t
J2,3 2.2
5.08 dd
J3,4 4.0
4.31 m
J1,2 1.2
5.56 dd
J2,3 1.2
5.62 dd
J3,4 2.5
4.79 m
J1,2 4.2
3.88 dd
J2,3 5.8
4.27 dd
J3,4 7.2
3.91 m
2
3
4
5a
4.79 m
3.97 m
4.31 m
5b
4.79 m
3.97 m
3.97 m
4.31 m
Other signals
CH3 1.36 t
J 7.5
CH3 1.29 t
J 7.5
CH3 1.29 t
J 7.5
CH3 1.30 t
J 7.5
CH2 2.77 m
CH2 2.77 m
OH 2.19 m
CH2 2.69 m
OCH2Ph 4.58 2d
OAc 2.06;
2.09, s
CH2 2.68 m
OCH2Ph 4.55 2d

114
J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
120 resin (H+), filtered and the solvent evap-
orated. Filtration on silica (10:1 CH2Cl2–
(25 mL) was treated with nBu4NF (1 M in
THF, 6.3 mL, 6.3 mmol) for 15 min at rt.
After evaporation of the solvent, the crude
product was dried under high vacuum and
taken up in pyridine (2 mL) and acetic anhy-
dride (2 mL) for 4 h. Chromatography (3:1
petroleum ether–EtOAc) gave 8 as a slightly
yellow oil (965 mg, 77%); [h]2D5 +113° (c
1.05, CHCl3); H NMR data see Table 1; C
NMR (CDCl3): l 170.73, 170.13 (CꢀO),
159.42, 129.50, 129.21, 113.84 (Ar), 88.00 (C-
1), 87.49 (C-2), 79.68 (C-4), 77.72 (C-3),
71.70 (OCH2Ar), 63.13 (C-5), 55.27 (OCH3),
25.42 (CH2S), 20.91, 20.80 (CH3CO), 14.85
MeOH) gave ethyl 1-thio-a- -arabino-
D
furanoside (4) (2.09 g). This compound was
taken up in pyridine (30 mL) and treated with
TIPS chloride (4 mL, 12.50 mmol) at rt. Af-
ter 3 h at rt, the solvent was evaporated and
the last traces of pyridine were removed by
coevaporation with toluene. The residue was
taken up in EtOAc and washed once with
water before drying on MgSO4. Chromato-
graphy (3:1 petroleum ether–EtOAc) gave 5
as a colorless oil (4.50 g, 65%); [h]2D5 +71° (c
1
13
1
13
1.26, CHCl3); H NMR data see Table 1; C
NMR (CDCl3): l 87.99 (C-1), 82.08 (C-2),
80.04 (C-4), 76.23 (C-3), 61.04 (C-5), 25.56
(CH2S), 17.41, 17.29, 17.07, 17.04, 16.97
(iPr), 15.02 (CH3CH2S), 13.45, 13.09, 12.74,
12.51 (iPr); ISMS: m/z 454 [M−1+Na].
Anal. Calcd for C19H40O5SSi2: C, 52.25; H,
9.23. Found: C, 51.81; H, 9.18.
(CH3CH2S);
ISMS:
m/z
420
[M−
1+Na]. Anal. Calcd for C19H26O7S: C,
57.26; H, 6.58. Found: C, 57.44; H, 6.61.
Methyl 3,5-di-O-acetyl-i- -arabinofuran-
D
oside (9).—A solution of 8 (650 mg, 1.63
mmol), MeOH (80 mL, 1.97 mmol) in CH2Cl2
,
(30 mL) was treated with activated 3 A
molecular sieves for 20 min before addition of
DDQ (555 mg, 2.44 mmol) at 0 °C. The reac-
tion mixture was stirred at rt for 6 h,
quenched with an aq solution of ascorbic
acid (0.7%), citric acid (1.3%) and NaOH
(0.9%) (20 mL) according to Ogawa’s proce-
dure [7]. The mixture was filtered on Celite
and the filtrate was washed with aq NaHCO3
and satd NaCl. Evaporation and filtration of
the residue on silica (3:1 petroleum ether–
EtOAc) gave the intermediate mixed acetal
contaminated with some starting material
(550 mg).
Ethyl 2-O-(4-methoxybenzyl)-3,5-O-(1,1,3,3-
tetraisopropyldisiloxane - 1,3 - diyl) - 1 - thio - h-
D-arabinofuranoside (6).—A solution of 5
(3.75 g, 8.58 mmol) in MeCN (25 mL) was
cooled to 0 °C before addition of 2-tert-
butylimino-2-diethylamino-1,3-dimethylperhy-
dro-1,3,2-diazaphosphorine (5 mL, 17.27
mmol) and a solution of 4-methoxybenzyl
bromide (9.10 g, 42.30 mmol) in 10 mL of
MeCN. The reaction medium was kept for 15
min at 0 °C and then 4 h at rt before concen-
tration. The residue was extracted with
EtOAc–satd NaHCO3 and the organic phase
was dried on MgSO4. Chromatography (25:1
petroleum ether–EtOAc) gave 6 as a slightly
yellow oil (2.26 g, 59%); [h]2D5 +60° (c 1.01,
CHCl3); 1H NMR data see Table 1; 13C
NMR (CDCl3): l 159.21, 129.87, 129.41,
113.67 (Ar), 89.07 (C-2), 86.42 (C-1), 79.67
(C-4), 76.05 (C-3), 72.30 (OCH2Ar), 61.11
(C-5), 55.26 (OCH3), 25.46 (CH2S), 17.44,
17.32, 17.09, 17.05, 16.98 (iPr), 14.91
(CH3CH2S), 13.49, 13.11, 12.82, 12.50 (iPr);
ISMS: m/z 557 [M+1]. Anal. Calcd for
C27H48O6SSi2: C, 58.23; H, 8.69. Found: C,
58.39; H, 8.66.
This mixture was taken up in CH2Cl2 (35
,
mL) and stirred with 3 A molecular sieves
and IDCP (600 mg, 1.28 mmol) for 20 min at
rt. A solution of Me3SiOTf (1 M in toluene,
107 mL, 0.107 mmol) was added and the re-
action left to proceed for 2 h. After neutral-
ization with triethylamine, the salts were
filtered on Celite and the filtrate evaporated.
Chromatography (1:1 petroleum ether–
EtOAc) gave 9 as a slightly yellow oil (200
1
mg, 51%); [h]2D5 −95° (c 1.04, CHCl3); H
NMR data see Table 2; 13C NMR (CDCl3):
l 170.98, 170.68 (CꢀO), 102.33 (C-1), 79.37
(C-3), 79.23 (C-4), 76.49 (C-2), 65.38 (C-5),
55.52 (OCH3), 20.85, 20.81 (CH3CO); ISMS:
m/z 271 [M+Na]. Anal. Calcd for C10H16O7:
C, 48.38; H, 6.50. Found: C, 48.71; H, 6.46.
Ethyl 3,5-di-O-acetyl-2-O-(4-methoxybenz-
yl)-1-thio-h-
tion of 6 (1.75 g, 3.14 mmol) in dry THF
D
-arabinofuranoside (8).—A solu-

J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
115
Table 2
1H NMR chemical shifts (l), apparent multiplicities and coupling constants in Hz for b-
D
-arabinofuranoside derivatives 9–13
H
1
9
10
11
12
13
4.90 d
J1,2 4.5
4.27 ddd
J2,3 6.2
4.73 d
J1,2 4.0
3.92 m
4.80 d
J1,2 4.5
4.14 m
4.71 d
4.62 d
J1,2 4.2
4.07 dd
J2,3 6.5
J1,2 4.5
4.07 dd
J2,3 7.5
2
J2,OH 8.5
5.05 dd
J3,4 5.0
4.11 ddd
J4,5a 3.2
J4,5b 7.0
4.16 dd
J5a,5b 11.0
3
4
3.92 m
4.05 m
4.19 dd
J3,4 5.2
4.05 m
4.26 dd
J3,4 6.0
4.04 m
3.76 ddd
J4,5a 4.0
J4,5b 7.0
3.54 dd
3.89 ddd
J 4.5, 5.0,
7.5
3.77 d
J5a,4 5.0
5a
3.73 d
J5a,4 6.2
3.56 dd
J5a,4 5.5
J5a,5b 11.5
J5a,5b 12.0
3.69 dd
5b
4.37 dd
3.66 dd
3.77 d
3.73 d
J5b,4 3.5
Other signals
OCH3 3.48 s
OAc 2.09,
2.12, s
OCH3 3.40 s
OCH3 3.36 s
CH3 1.05 s
OCH3 3.26 s
CH3 1.04 s
OCH3 3.40 s
OH 2.15 m
OH 2.80 d
Methyl
2,3-di-O-benzyl-i-
D
-arabinofura-
petroleum ether–EtOAc) gave methyl 2,3-di-
noside (13).—Deacetylation of 9 (220 mg, 0.88
mmol) was carried out as described above for
the preparation of 4. Chromatography (10:1
O-benzyl-5-O-tert-butyldiphenylsilyl-b- -ara-
D
binofuranoside (12) as a colorless oil (110 mg,
87%); [h]2D5 −24° (c 1.04, CHCl3); H NMR
1
CH2Cl2 –MeOH) gave methyl b-
D-arabino-
data see Table 2; 13C NMR (CDCl3): l 135.58,
129.68, 128.37, 128.31, 128.18, 127.90, 127.68,
127.58, 126.86 (Ar), 101.51 (C-1), 84.29, 83.32,
82.04 (C-2, C-3, C-4), 72.52, 72.37 (OCH2Ph),
66.01 (C-5), 54.93 (OCH3), 29.70, 26.82 (tBu).
Compound 12 (150 mg, 0.257 mmol) was
treated in THF (3 mL) with nBu4NF (1 M in
THF, 290 mL, 0.29 mmol) for 40 min at rt.
Work-up and chromatography (2:1 petroleum
ether–EtOAc) gave 13 as a slightly yellow oil
furanoside (10) (115 mg, 80%); [h]2D5 −114° (c
1
1.05, MeOH), lit. −117° [35]; H NMR data
see Table 2. Compound 10 (88 mg, 0.54
mmol) in DMF (4 mL) was treated at rt with
imidazole (48 mg, 0.70 mmol) and tert-
butyldiphenylsilyl chloride (150 mL, 0.58
mmol) and the mixture was heated at 60 °C
for 1 h. Work-up and chromatography (1:2
petroleum ether–EtOAc) gave methyl 5-O-
1
tert-butyldiphenylsilyl-b-
D
-arabinofuranoside
(65 mg, 74%); [h]2D5 −38° (c 1.03, CHCl3); H
(11) as a colorless oil (140 mg, 65%); [h]D25
NMR data see Table 2; C NMR (CDCl3): l
13
1
−48° (c 1.04, CHCl3); H NMR data see
137.91, 137.47, 128.42, 128.18, 128.01, 127.81,
127.76, 125.84 (Ar), 101.83 (C-1), 84.31 (C-2),
82.22 (C-4), 80.97 (C-3), 72.67, 72.58
(OCH2Ph), 63.99 (C-5), 55.70 (OCH3). Anal.
Calcd for C20H24O5: C, 69.75; H, 7.02. Found:
C, 69.56; H, 7.21.
13
Table 2; C NMR (CDCl3): l 133.55, 133.20,
129.81, 129.77, 127.74, 125.84 (Ar), 101.75
(C-1), 81.82 (C-4), 78.04 (C-3), 65.03 (C-5),
55.37 (OCH3), 26.80, 19.24 (tBu).
A solution of 11 (87 mg, 0.216 mmol) and
benzyl bromide (56 mL, 0.47 mmol) in DMF
(3 mL) was treated at 0 °C with NaH (60%
dispersion in mineral oil, 32 mg, 0.86 mmol).
The reaction was left at rt for 4 h, quenched
with MeOH and a few drops of 1 M HCl
before concentration. Chromatography (5:1
Methyl 2,3-di-O-acetyl-5-(3,5-di-O-acetyl-
i-D
-arabinofuranosyl)-h- -arabinofuranoside
D
(14).—A mixture of 8 (50 mg, 0.125 mmol)
and methyl 2,3-di-O-acetyl-a- -arabinofuran-
oside (32.5 mg, 0.132 mmol) was stirred with
D
,
activated 4 A molecular sieves in CH2Cl2 for

116
J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
20 min before addition of DDQ (42.5 mg,
0.187 mmol) at 0 °C. The reaction was left 3 h
at rt and treated as described above for the
preparation of 9. Chromatography (4:1 then
1:1 petroleum ether–EtOAc) gave the inter-
mediate acetal as a colorless oil (57 mg, 71%).
This oil was treated as above in CH2Cl2 (1.6
mL) containing IDCP (48 mg, 0.102 mmol)
CD3OD): l 138.61, 138.32, 138.23, 138.01,
137.95, 128.17, 127.94, 127.52, 127.43, 127.23,
121.47, 83.02, 81.25, 80.64, 80.47, 75.65, 75.50,
31
74.68, 72.18; P NMR (9:1 CDCl3–CD3OD):
l 8.80; ESMS: m/z 693 [M−1]. Anal. Calcd
for C41H43O8P: C, 70.88; H, 6.24. Found: C,
70.72; H, 6.08.
1 -1,2,4,5,6-Penta-O-benzyl-myo-inositol 3-
D
,
hydrogen phosphonate (18).—Starting from 17
(150 mg, 0.24 mmol) and following the proce-
dure described above for the preparation of
16, compound 18 was obtained as a white
foam (95 mg, 56%); [h]2D5 −23° (c 1.65,
CHCl3); for NMR data, see 16; ESMS: m/z
693 [M−1]. Anal. Calcd for C41H43O8P: C,
70.88; H, 6.24. Found: C, 70.82; H, 6.07.
and 4 A molecular sieves. Glycosylation, pro-
moted with Me3SiOTf (8.5 mL, 0.008 mmol)
was over in 45 min at rt. Work-up and chro-
matography (1:2 petroleum ether–EtOAc)
gave 14 as a yellow oil (29 mg, 52% from 8);
[h]2D5 −22° (c 1.40, CHCl3); 1H NMR
(CDCl3): l 5.12 (d, 1 H, J1%,2% 4.5 Hz, H-1%),
5.10 (d, 1 H, J2,3 1.2 Hz, H-2), 5.09 (dd, 1 H,
J2%,3% 6.5, J3%,4% 5.0 Hz, H-3%), 5.03 (dd, 1 H, J3,4
4.5 Hz, H-3), 4.93 (s, 1 H, H-1), 4.37 (dd, 1 H,
J5a%,5b% 11.2, J5a%,4% 4.0 Hz, H-5a%), 4.31 (dd, 1 H,
H-2%), 4.20 (dd, 1 H, J5b%,4% 7.5 Hz, H-5b%), 4.11
(dd, 1 H, J5a,5b 10.8, J5a,4 3.0 Hz, H-5a),
4.20–4.10 (m, 2 H, H-4 and H-4%), 3.76 (dd, 1
H, J5b,4 4.2 Hz, H-5b), 3.40 (s, 3 H, OMe),
2.09–2.12 (4s, 12 H, OAc); 13C NMR
(CDCl3): l 170.66, 170.47, 169.79 (CꢀO),
106.71 (C-1), 101.54 (C-1%), 82.01, 81.35,
79.41, 78.97, 77.19, 67.19, 65.48 (C-5, C-5%),
54.93 (OCH3). Anal. Calcd for C19H28O13: C,
49.14; H, 6.08. Found: C, 48.78; H, 5.92.
Methyl
2,3-di-O-benzyl-5-(1 -1,2,4,5,6-
L
penta-O-benzyl-myo-inositol 3-phosphate) i-
D-
arabinofuranoside (19).—A solution of 16 (20
mg, 0.028 mmol) and 13 (8.8 mg, 0.025 mmol)
in anhyd pyridine (0.75 mL) was stirred for 20
,
min with activated 4 A molecular sieves before
addition of pivaloyl chloride (9.5 mL, 0.077
mmol). After 20 h at rt, a solution of iodine
(13 mg, 0.051 mmol) in 98% aq pyridine (0.3
mL) was added and the mixture left for an
additional 1 h before filtration on Celite. The
filtrate was diluted with CH2Cl2 and washed
with satd sodium thiosulfate. Chromatogra-
phy (15:1 CH2Cl2–MeOH) gave 19 as a color-
less oil (17.5 mg, 67%); [h]2D5 −4.8° (c 0.87,
1 -1,2,4,5,6-Penta-O-benzyl-myo-inositol 3-
L
hydrogen phosphonate (16).—A solution of
imidazole (220 mg, 3.23 mmol) in anhyd
toluene (2.7 mL) was treated at 0 °C with PCl3
(62 mL, 0.71 mmol in 0.65 mL of toluene) and
Et3N (220 mL, 1.58 mmol in 0.65 mL of
toluene). After 10 min, the temperature was
lowered to −5 °C and a solution of 15 (150
mg, 0.24 mmol) in 4.5 mL of toluene was
added dropwise over 1 h. The reaction mix-
ture was left for 1 h and allowed to warm to
15 °C before quenching with aq pyridine (4:1
pyridine–water, 35 mL); CH2Cl2 extraction
and chromatography (8:1 CH2Cl2–MeOH)
gave 16 as a white foam (97 mg, 57%); [h]D25
1
CHCl3); H NMR (9:1 CDCl3–CD3OD): l
7.35–7.03 (m, 35 H, Ar), 4.87–4.41 (m, 15 H,
CH2Ph, H-1), 4.37 (s, 1 H, H-2%), 4.05–3.84
(m, 8 H, H-1%, H-4%, H-6%, H-2, H-3, H-4,
H-5), 3.35 (dd, 1 H, J3%,4% 9.5, J3%,2% 2.0 Hz,
H-3%), 3.28 (t, 1 H, J5%,6%ꢀJ5%,4% 9.5 Hz, H-5%),
3.17 (s, 3 H, OCH3); 13C NMR (9:1 CDCl3–
CD3OD): l 138.32, 137.67, 137.24, 128.48,
128.16, 127.94, 127.55, 127.40 (Ar), 101.69
(C-1), 83.82, 82.98, 82.70, 81.40, 80.69, 80.60,
80.33, 75.72, 74.81, 72.37, 72.20, 72.03, 67.97,
54.78; 31P NMR (9:1 CDCl3–CD3OD, 303 K):
l 0.67; ESMS: m/z 1036 [M−1].
1
+22° (c 1.49, CHCl3); H NMR (9:1 CDCl3–
Methyl 2,3-di-O-benzyl-5-(1
D
-1,2,4,5,6-pen-
CD3OD): l 7.76–6.90 (m, 25 H, Ar), 6.91 (d,
1 H, JP,H 642 Hz, H–P), 7.94–4.72 (m, 8 H,
CH2Ph), 4.63, 4.58 (2d, 2 H, J 11.0 Hz,
CH2Ph), 4.23 (br t, 1 H, J1,2 2.2, J2,3 2.2 Hz,
H-2), 4.19–3.95 (m, 3 H, H-4, H-5, H-6), 3.44
(m, 2 H, H-1, H-3); 13C NMR (9:1 CDCl3–
ta-O-benzyl-myo-inositol 3-phosphate)-i- -
D
arabinofuranoside (20).—Starting from 18 (20
mg, 0.028 mmol) and 13 (8.8 mg, 0.025 mmol)
and following the procedure described
above for the preparation of 19, compound 20
was obtained (18 mg, 69%); [h]2D5 −26°

J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
117
Table 3
1H NMR chemical shifts (l), apparent multiplicities, coupling constants in Hz, and 13C NMR chemical shifts (l) for compounds
1 and 2 (D2O, 303 K)a
H-1
H-2
H-3
H-4
H-5
H-5
1
2
4.90, d
J1,2 4.2
4.89, d
J1,2 4.5
4.14, dd
J2,3 7.5
4.14, dd
J2,3 8.0
4.09, dd
J3,4 6.0
4.09, m
J3,4 6.0
4.01, m
4.00, m
4.07, m
4.04, m
3.95, m
3.97, m
H-1%
H-2%
H-3%
H-4%
H-5%
H-6%
1
3.94, ddd
J1%,2% 2.7
J1%,P 11
4.25, t
J2%,3% 2.7
3.54, dd
J3%,4% 9.5
3.64, t
J4%,5% 9.5
3.31, t
J5%,6% 9.5
3.74, t
J6%,1% 9.5
2
3.95, ddd
J1%,2% 2.8
3.98, t
J1%,2% 2.2
J1%,P 9.6
4.25, t
3.53, dd
J3%,4% 9.5
3.58, d
3.63, t
J4%,5% 9.5
3.67, t
3.31, t
J5%,6% 9.5
3.35, t
3.74, t
J6%,1% 9.5
3.77, t
J2%,3% 2.8
4.28, s
J2%,3% 2.2
LAM
J3%,4% 9.2
J4%,5% 9.2
J5%,6% 9.6
J6%,1% 9.6
C-1
C-2
C-3
C-4
C-5
1
2
103.45
103.18
77.15
77.18
75.38
75.42
81.52
81.51
67.50
67.46
C-1%
C-2%
C-3%
C-4%
C-5%
C-6%
1
2
77.29
77.32
77.37
72.19
72.28
72.20
71.62
71.82
71.61
73.21
73.28
73.13
74.89
74.99
74.89
72.35
72.47
72.24
LAM
a Values for the LAM are taken from Ref. [5].
(c 1.11, CHCl3); 1H NMR (9:1 CDCl3–
CD3OD): l 7.35–7.04 (m, 35 H, Ar), 4.88–
4.42 (m, 15 H, CH2Ph, H-1), 4.37 (s, 1 H,
H-2%), 4.04–3.90 (m, 8 H, H-1%, H-4%, H-6%,
H-2, H-3, H-4, H-5), 3.36 (dd, 1 H, J3%,4% 9.5,
J3%,2% 2.0 Hz, H-3%), 3.29 (t, 1 H, J5%,6%ꢀJ5%,4% 9.5
(OH)2–C for 8 h. The catalyst was filtered
on Celite and the residue was chro-
matographed on silica (1:2:1 EtOAc–
MeOH–H2O) to give 1 as a white foam (6.5
1
mg, 56%); for H NMR and 13C NMR data,
see Table 3; 31P NMR (D2O, 303 K): l 5.52;
ESMS: m/z 405 [M−1].
13
Hz, H-5%), 3.18 (s, 3 H, OCH3); C NMR (9:1
Methyl i-
D
-arabinofuranoside 5-(1 -myo-
L
CDCl3-CD3OD): l 138.92, 138.41, 138.26,
138.14, 137.82, 137.59, 137.18, 128.27, 128.13,
128.01, 127.93, 127.85, 127.53, 127.35, 127.29,
127.11 (Ar), 101.54 (C-1), 83.78, 82.91 (C-5%),
82.26, 81.35, 80.88, 80.46, 80.34 (C-3%), 76.37
(C-2%), 76.11, 75.81, 75.70, 74.77, 72.31, 72.10,
71.98, 67.72 (C-5), 54.70 (OCH3); 31P NMR
(9:1 CDCl3–CD3OD, 303 K): l 0.67; ESMS:
m/z 1036 [M−1].
inositol 1-phosphate) (2).—Hydrogenolysis of
20 (30 mg, 0.029 mmol), as above, gave 2
1
as a white foam (10.5 mg, 79%); for H
NMR and 13C NMR data, see Table 3; 31P
NMR (D2O, 303 K): l 5.52; ESMS: m/z 405
[M−1].
Acknowledgements
Methyl i-
D
-arabinofuranoside 5-(1 -myo-
D
inositol 1-phosphate) (1).—Compound 19 (30
mg, 0.029 mmol) in 1:2:1 EtOAc–MeOH–
H2O (0.5 mL) was hydrogenolyzed on Pd-
We thank Patricia Constant, IPBS-CNRS,
for performing the TNF-a production tests on
our products.

118
J. De´sire´, J. Prandi / Carbohydrate Research 317 (1999) 110–118
[22] S. Kaneko, Y. Kawabata, T. Ishii, Y. Gama, I.
References
Kusakabe, Carbohydr. Res., 268 (1995) 307–311.
[23] P. Garegg, I. Kvarnstro¨m, A. Niklasson, G. Niklasson,
S.C.T. Svensson, J. Carbohydr. Chem., 12 (1993) 933–954.
[24] N. Kornblum, R.A. Smiley, R.K. Blackwood, D.C.
Iffland, J. Am. Chem. Soc., 77 (1955) 6269–6280.
[25] B.S. Sproat, B. Beijer, A. Iribarren, Nucleic Acids Res., 18
(1990) 41–49.
[26] C.H. Gotfredsen, J.P. Jacobsen, J. Wengel, Tetrahedron
Lett., 35 (1994) 6941–6944.
[27] M. Grøtli, M. Douglas, B. Beijer, R. Eritja, B. Sproat,
Biorg. Med. Chem. Lett., 7 (1997) 425–428.
[28] R.U. Lemieux, A.R. Morgan, Can. J. Chem., 43 (1965)
2190–2198.
[29] G.H. Veeneman, S.H. Van Leeuwen, H. Zuurmond, J.H.
van Boom, J. Carbohydr. Chem., 9 (1990) 783–796.
[30] P. Smid, G.A. de Ruiter, G.A. van der Marel, F.M.
Rombouts, J.H. van Boom, J. Carbohydr. Chem., 10
(1991) 833–849.
[31] P.A.J. Gorin, M. Mazurek, Can. J. Chem., 53 (1975)
1212–1223.
[32] R.G.S. Ritchie, N. Cyr, B. Korsch, H.J. Koch, A.S. Perlin,
Can. J. Chem., 53 (1975) 1424–1433.
[33] R.C. Beier, B.P. Mundy, J. Carbohydr. Chem., 3 (1984)
253–266.
[1] D. Chatterjee, K.-H. Khoo, Glycobiology, 8 (1998) 113–
120.
[2] T.I.A. Roach, C.H. Barton, D. Chatterjee, J.M. Blackwell,
J. Immunol., 150 (1993) 1886–1896.
[3] A. Venisse, J.-J. Fournie´, G. Puzo, Eur. J. Biochem., 231
(1995) 440–447.
[4] S. Prinzis, D. Chatterjee, P.J. Brennan, J. Gen. Microbiol.,
139 (1993) 2649–2658.
[5] M. Gilleron, N. Himoudi, O. Adam, P. Constant, A.
Venisse, M. Rivie`re, G. Puzo, J. Biol. Chem., 272 (1997)
117–124.
[6] Y. Ito, T. Ogawa, Angew. Chem., Int. Ed. Engl., 33 (1994)
1765–1767.
[7] A. Dan, Y. Ito, T. Ogawa, J. Org. Chem., 60 (1995)
4680–4681.
[8] H.B. Mereyala, S. Hotha, M.K. Gurjar, Chem. Commun.
(Cambridge), (1998) 685–686.
[9] B.A. Wolucka, M.R. McNeil, E. de Hoffmann, T. Choj-
nacki, P.J. Brennan, J. Biol. Chem., 269 (1994) 23328–
23335.
[10] R.E. Lee, K. Mikus' ova´, P.J. Brennan, G.S. Besra, J. Am.
Chem. Soc., 115 (1995) 11829–11832.
[11] R.E. Lee, P.J. Brennan, G.S. Besra, Glycobiology, 7 (1997)
1121–1128.
[34] A.S. Serianni, R. Barker, J. Org. Chem., 49 (1984) 3292–
3300.
[35] C.T. Bishop, F.P. Cooper, Can. J. Chem., 41 (1963)
2743–2758.
[36] W.J. Hennen, H.M. Sweers, Y.-F. Wang, C.-H. Wong, J.
Org. Chem., 53 (1988) 4939–4945.
[12] F. Barresi, O. Hindsgaul, J. Am. Chem. Soc., 113 (1991)
9376–9377.
[13] F. Barresi, O. Hindsgaul, Synlett, (1992) 759–761.
[14] G. Stork, G. Kim, J. Am. Chem. Soc., 114 (1992) 1087–
1088.
[37] C.E. Dreef, M. Douwes, C.J.J. Elie, G.A. van der Marel,
J.H. van Boom, Synthesis, (1991) 443–447.
[38] C. Pratt, Y. Liu, T. Chu, K. Melkonian, B.E. Tropp, R.
Engel, Can. J. Chem., 70 (1992) 2135–2141.
[39] P.J. Garegg, T. Regberg, J. Stawin˜ski, R. Stro¨mberg,
Chem. Scr., 25 (1985) 280–282.
[40] P.J. Garegg, I. Lindh, T. Regberg, J. Stawin˜ski, C.
Henrichson, Tetrahedron Lett., 27 (1986) 4055–4058.
[41] I. Lindh, J. Stawin˜ski, J. Org. Chem., 54 (1989) 1338–
1342.
[15] M. Bols, J. Chem. Soc., Chem. Commun., (1992) 913–914.
[16] M. Bols, Tetrahedron, 44 (1993) 10049–10060.
[17] C. Krog-Jensen, S. Oscarson, J. Org. Chem., 61 (1996)
4512–4513.
[18] C. Krog-Jensen, S. Oscarson, J. Org. Chem., 63 (1998)
1780–1784.
[19] R.K. Ness, H.G. Fletcher, Jr., J. Am. Chem. Soc., 80
(1958) 2007–2010.
[20] W.T. Markiewicz, J. Chem. Res. (S), (1979) 24–25.
[21] S. Czernecki, T. Le Diguarher, Synthesis, (1991) 683–686.
.