Syntheses of the L-manno and some other analogs of the terminal determinants of the O-PS of Vibrio cholerae O:1

Article abstract of DOI:10.1016/S0008-6215(00)00265-2

Analogs of the methyl α-glycosides of the terminal residues of the O-specific polysaccharides (O-PS) of Vibrio cholerae O:1, serotype Inaba and Ogawa, have been prepared as probes to study their interaction with anti V. cholerae O:1 antibodies. They differ from the termini of the respective O-PSs in anomeric or absolute configuration of perosamine, position of the O-methyl group in d-perosamine, and nature of the N-acyl side chain.

Full text of DOI:10.1016/S0008-6215(00)00265-2

www.elsevier.nl/locate/carres
Carbohydrate Research 330 (2001) 7–20
Syntheses of the -manno and some other analogs of the
L
terminal determinants of the O-PS of Vibrio cholerae O:1
Emmanuel Poirot, Xiaodong Zhang, Noel F. Whittaker, Pavol Kova´cˇ*
National Institutes of Health, Building 8, Room B1A24, Bethesda, MD 20892-0815, USA
Received 24 July 2000; accepted 14 September 2000
Abstract
Analogs of the methyl a-glycosides of the terminal residues of the O-specific polysaccharides (O-PS) of Vibrio
cholerae O:1, serotype Inaba and Ogawa, have been prepared as probes to study their interaction with anti V.
cholerae O:1 antibodies. They differ from the termini of the respective O-PSs in anomeric or absolute configuration
of perosamine, position of the O-methyl group in
D-perosamine, and nature of the N-acyl side chain. © 2001
Elsevier Science Ltd. All rights reserved.
Keywords: Vibrio cholerae O:1; Yersinia enterocolitica; Brucella abortus; Perosamine
1. Introduction
dideoxy-4-(3-deoxy-
-mannopyranose
tetronamido)-a- -perosamine]. The two O-
L
-glycero-tetronamido)-a-
D
[4-(3-deoxy- -glycero-
L
Cholera is a serious, enteric infectious dis-
ease caused, inter alia, by Vibrio cholerae O:1
bacterial pathogens. A satisfactory vaccine for
the disease is not available. One of the ongo-
ing projects in this laboratory is aimed at
developing a potent immunogen for anti V.
cholerae O:1 antibodies that would provide
long-lasting protection against cholera. Our
approach involves conjugation of synthetic,
linker-equipped fragments of the O-specific
antigen (O-specific polysaccharide, O-PS) to a
carrier protein.
D
PSs differ in that the position O-2 of the
upstream, terminal perosamine group in the
Ogawa strain is methylated (Fig. 1). We have
V. cholerae O:1 is divided into two
serotypes, Inaba and Ogawa. The internal
parts of their O-PSs are identical and con-
sist1,2 of a chain of a-(1“2)-linked 4,6-
* Corresponding author. Tel.: +1-301-4963569; fax: +1-
301-4020589.
E-mail address: kpn@helix.nih.gov (P. Kova´cˇ).
Fig. 1.
0008-6215/01/$ - see front matter © 2001 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(00)00265-2

8
E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
studied3 binding of some monoclonal antibod-
ies for V. cholerae O:1 with synthetic frag-
ments of the O-PS and some analogs of the
terminal monosaccharide residues. We found,
among other things, that some antibodies
showed a remarkable tolerance to irregulari-
ties in the structure of ligands. For example,
O:1 O-PSs in the anomeric or absolute
configuration of perosamine, the nature of the
amide group, or in the position of the methyl
group in the
D-perosamine moiety.
To prepare the b analogs of the monosac-
charide determinants of the O-PS of V.
cholerae O:1, serotype Inaba (6) and Ogawa
(9), we applied our method of glycosylation
via locked anomeric configuration,8,9 which
involves the use of 1,2-O-stannylene acetals as
intermediates. We showed previously10 that
the undesirable epimerization that often ac-
companies the formation of such compounds
from free sugars could be avoided by position-
ing an alkyl group at O-3 in the synthon.
Thus, diol 3, obtained from 1 through the
acetate 2,11–13 was converted to the corre-
sponding stannylene acetal, which was then
treated8,9 with methyl trifluoromethanesul-
fonate (methyltriflate), to give stereospecifi-
cally the corresponding b-glycoside (4) in 82%
yield. Subsequent methylation of 4 gave 7.
The foregoing azides were converted to the
corresponding amines (5 and 8 respectively),
changing the
L
configuration of the
-series
tetronamido side chain for that of the
D
did not result in cessation of binding of the
monoclonal antibodies specific to V. cholerae
O:1, serotype Ogawa. Another interesting
finding was that the monosaccharide ligand
belonging to the Ogawa series that was deoxy-
genated at position 4% showed higher affinity
for the antibodies studied than the ligand
reflecting the structure of the end-group in the
natural Ogawa O-PS. To learn more about the
structural requirements for binding, and per-
haps reveal structural elements that would
increase binding, we have now synthesized a
series of new analogs of the upstream terminal
monosaccharides of the two serotypes.
In addition to the O-PS of V. cholerae O:1,
variously N-acylated perosamine is the key
constituent of O-specific polysaccharides of
many other bacterial pathogens. There, the
O-PS differ from that of V. cholerae O:1
mainly in the nature of the N-acyl side chain.
In this context, it is interesting to note that N-
which were treated with 3-deoxy- -glycero-
L
tetronolactone, to give the target glycosides 6
and 9 (Scheme 1). The intermediate com-
pound 2, previously obtained amorphous,14
was now obtained crystalline.
Probing the importance of the location of
the methyl group at O-2 for binding of the
monosaccharide determinant of the O-PS of
V. cholerae O:1, serotype Ogawa, with their
homologous antibodies required the 3-O-
methyl derivative 15. It was obtained from
diol 1011–13 as starting material. Peters and
Bundle15 have shown that benzylation of 10
under phase-transfer catalysis gave 2-O-benzyl
ether as the major product. When 10 was
treated with iodomethane under the same con-
ditions, similar regioselectivity of alkylation
was observed, yielding the crystalline, hitherto
unknown 2-methyl ether 12 as the main
product. In addition, a small amount of the
2,3-di-O-methyl derivative 13 and the 3-O-
methyl derivative 11 were also obtained
(Scheme 2). The use of this method of methyl-
ation was preferred, compared to the one
yielding preferentially the corresponding 3-
alkyl derivative,12 as compound 12 was
3-hydroxypropionyl-a-
D
-perosamine and N-3-
hydroxypropionyl - 2 - O - methyl - a -
D-
perosamine were identified as constituents of
the lipopolysaccharide from Vibrios serogroup
1875, referred to as Original and Variant,
respectively.4,5 On the other hand, the O-
polysaccharide chain from V. cholerae O-766
and O-1447 are homopolymers of, respec-
tively, N-2-hydroxy-L- and
D
-propionyl-a- -
L
perosamine. Thus, some of the compounds
herein described for the first time are related
to structural features of important epitopes in
the above O-polysaccharides and, as such, can
be useful for immunochemical studies also in
those antigen–antibody systems.
2. Results and discussion
The synthetic ligands reported here differ
from the end-groups in the natural V. cholerae

E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
9
Scheme 1.
needed in much larger amount during this
work. Compound 15 was then readily synthe-
sized from 11 via amine 14.
Many analogs here prepared (18–27) differ
from the regular constituents of the O-PS of
V. cholerae O:1 in the acylamido group.
They were obtained from amines 16 and 17 by
N-acylation with various acylating reagents,
some of which were available commercially.
The hitherto unknown acid 38 was prepared
by acetylation of the hemicalcium salt of the
parent, unprotected acid. In many cases, chro-
matography of crude products of N-acylation
readily yielded pure materials. When purifica-
tion of desired products was difficult, crude
products of such couplings were acetylated
with Ac2O in pyridine, and the per-O-acetates
thus obtained were purified by chromatogra-
phy and crystallization. Subsequent deacetyla-
tion then gave the pure, desired substances
(c.f., for example the sequence 5“crude 6“
28“6).
Each of the reactions of 38 with amines 16
and 17 gave two products. To aid separation
and purification, crude products of these reac-
tions were processed as described above, and
the fully acetylated derivatives were isolated
by chromatography. The reaction of 16
yielded compounds 32 and 34, the former
largely predominating. When examined by
CIMS, these compounds produced the same
pseudomolecular ion peaks, and their NMR
spectra were very similar, indicating that these
materials were a pair of diastereoisomers.

10
E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
Scheme 2.
Compounds 32 and 34 were fully character-
ized and identified as the - and -glycero
L
D
isomers, respectively, based on the optical ro-
tation data. The specific optical rotation we
found for the commercial
calcium salt monohydrate suggested that the
material contained a small proportion of the
L
-glyceric acid hemi-
D
enantiomeric form, which explains the forma-
tion of diastereoisomers described above. Sim-
ilarly, the predominating product of the
reaction of 17 with the same N-acylation
reagent, followed by acetylation, was the
L
-
compound 33.
The title
L
analogs 45 and 46 were obtained
-glycero-
tetronolactone of amines 4116 and 44, which
were obtained from methyl a- -rhamnopyrano-
by N-acylations with 3-deoxy-
L
L
side17 following synthetic strategies devel-
oped,11–13,18 for the corresponding
analogs.
D
3. Experimental
Optical rotations were measured at 25 °C
with a Perkin–Elmer model 341 automatic
polarimeter. All reactions were monitored by
thin-layer chromatography (TLC) on silica
gel-coated glass slides (Whatman or Anal-
tech). Detection was effected by charring with
5% H2SO4 in EtOH and, when applicable, by
UV light. Preparative chromatography was
performed by gradient elution from columns

E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
11
resolved mixture of anomeric acetates
obtained as described above, was deacetylated
(Zemple´n) to give 3 in virtually theoretical
yield, mp 135.5–136.5 °C (from EtOH–Et2O),
[h]D +95° (c 0.75, MeOH). 1H NMR
(CDCl3–CD3OD): l 5.11 (d, 1 H, J1,2 1.5 Hz,
H-1), 4.72, 4.67 (2 d, 2 H, CH2Ph), 4.00 (dd, 1
H, J2,3 3.0 Hz, H-2), 3.82–3.74 (m, 2 H,
H-3,5), 3.42 (t, 1 H, J 10.1 Hz, H-4), 1.30 (d,
3 H, J5,6 6.0 Hz, H-6). Before deuterium ex-
of Silica Gel 60 (E. Merck, particle size
0.035–0.070 mm) using at the onset of devel-
opment a solvent mixture slightly less polar
than that used for TLC. NMR spectra were
1
obtained at 300 MHz for H and 75 MHz for
13C with a Varian Mercury spectrometer. As-
signment of signals were made by first-order
analysis, homonuclear decoupling and/or
homo- and heteronuclear two-dimensional
correlation spectroscopy. Chemical ionization
mass spectra (CIMS, positive and negative
ion) were obtained on a Finnigan 4000
quadrupole mass spectrometer with NH3
reagent gas. Electron-impact ionization spec-
tra were obtained with a VG 70E mass spec-
trometer. Palladium-on-charcoal (5%) catalyst
(ESCAT 103) was purchased from Engelhart
Industries. The use of this catalyst largely
eliminated problems encountered when prod-
ucts of debenzylation and amines are formed
1
change, H NMR (CDCl3) showed the signal
for H-1 as a doublet of doublets, and the
signal for H-2 as a multiplet. The signals for
HO-1 (J1,OH 3.7 Hz) and HO-2 (J2,OH 1.8 Hz)
appear at l 2.56 and 2.44, respectively. 13C
NMR (CDCl3 –CD3OD): l 93.54 (C-1), 77.74
(C-3), 71.62 (CH2Ph), 67.34 (C-2), 66.21 (C-
5), 64.00 (C-4), 18.25 (C-6). Anal. Calcd for
C13H17N3O4: C, 55.91; H, 6.14; N, 15.05.
Found: C, 55.74; H, 6.14; N, 15.15.
Methyl 4-azido-3-O-benzyl-4,6-dideoxy-i-
-mannopyranoside (4).—A mixture of 3 (279
at the same time.19,20
-Lactic (2-hydroxypro-
L
D
pionic) acid was purchased from Sigma Chem-
ical Co., and glycolic acid was purchased from
mg, 1 mmol) and Bu2SnO (149 mg, 1 mmol)
in anhyd MeOH was stirred at 70 °C until a
clear soln was obtained (ꢀ1.5 h). CsF (0.91
g, 6 mmol) and toluene (5 mL) was added, the
mixture was concd, and the residue was kept
at 50 °C/133 Pa for 3 h. Methyl triflate (136
mL, 1.2 mmol) was added at −10 °C to a
suspension of the residue and molecular sieves
Aldrich Chemical Co. -Gyceric acid hemical-
L
cium salt monohydrate {[h]D −12.5° (c 4.6,
H2O), lit.21, [h]D −14.6° (c 5.0 H2O)}, b-pro-
piolactone and 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (EDAC)
were purchased from Fluka Chemical com-
pany. O - (7 - Azabenzotriazol - 1 - yl) - 1,1,3,3-
,
4 A (0.5 g) in DMF (10 mL), which had been
tetramethyluronium
hexafluorophosphate
stirred at rt for 2 h. Stirring was continued at
the same temperature until TLC (2:1 hexane–
acetone) showed that the reaction was essen-
tially complete (ꢀ30 min). Chromatography
gave pure 4 (241 mg, 82%), mp 88.5–89 °C
(HATU) was purchased from PerSeptive
Biosystems. The commercial products listed
were used without further purification. Solu-
tions in organic solvents were dried with an-
hyd Na2SO4 and concd at 40 °C/2 kPa.
1
(from 2-Pr2O), [h]D +17° (c 1, CHCl3). H
1,2-Di-O-acetyl-4-azido-3-O-benzyl-4,6-
NMR (CDCl3): l 4.76, 4.68 (2 d, 2 H, J 11.8
Hz, CH2Ph), 4.25 (d, 1 H, J1,2 1.1 Hz, H-1),
4.07 (m, 1 H, H-2), 3.52 (s, partially over-
lapped, OCH3), 3.49 (t, partially overlapped,
H-4), 3.40 (dd, 1 H, J2,3 3.0, J3,4 9.5 Hz, H-3),
3.14 (m, 1 H, H-5), 2.44 (bs, 1 H, OH), 1.45
dideoxy-h- -mannopyranose (2).—This com-
D
pound was prepared from the corresponding
methyl glycoside (1) as described.11,14 Crystal-
lization from 2-Pr2O–hexane gave pure 2, mp
47.5–48.5 °C [h]D +112.5° (c 0.7, CHCl3);
Ref.11 [h]D +89° (c 1, CHCl3) for the material
obtained in admixture with a small amount of
the b isomer; Ref.14, +111° (c 1.3, CH2Cl2)
for the previously reported amorphous, pure
a-acetate. Anal. Calcd for C17H21N3O6: C,
56.19; H, 5.83; N, 11.56. Found: C, 56.39; H,
5.86; N, 11.70.
13
(d, 3 H, J5,6 6.1 Hz, H-6); C NMR (CDCl3):
l 100.44 (C-1, JC-1,H-1 155.4 Hz), 79.81 (C-3),
71.31 (CH2Ph), 70.79 (C-5), 67.19 (C-2), 63.57
(C-4), 56.95 (OCH3), 18.41 (C-6); CIMS: m/z
293 ([M+18]+). Anal. Calcd for C14H19N3O4:
C, 57.33; H, 6.53; N, 14.33. Found: C, 57.49;
H, 6.53; N, 14.20.
4 - Azido - 3 - O - benzyl - 4,6 - dideoxy - h,i -
D-
Methyl
4-amino-4,6-dideoxy-i- -manno-
D
mannopyranose (3).—Compound 2, or the un-
pyranoside (5).—A soln of the azide 4 (1.43 g)

12
E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
Deacetylation of 28 (Zemple´n) gave 6 in
in EtOH (50 mL) was treated at rt and atmo-
spheric pressure with H2 in the presence of
ESCAT 103 (1 g) until TLC (4:1 CH2Cl2–
MeOH) showed that the reaction was complete
(24–48 h). The mixture was worked up con-
ventionally to give 5 in virtually theoretical
yield, mp 124–125.5 °C (from MeOH–hex-
virtually theoretical yield, mp 179.5–180 °C
1
(from EtOH), [h]D −84.5° (c 1.2, MeOH), H
NMR (D2O): l 4.51 (d, 1 H, J1,2 1.0 Hz, H-1),
4.27 (dd, 1 H, J2%,3%a 4.0, J2%,3%b 8.8 Hz, H-2%), 3.98
(dd, 1 H, J2,3 2.6 Hz, H-2), 3.77–3.74 (m,
partially overlapped, H-3,4), 3.72 (q, partially
overlapped, H-4%a,b), 3.57–3.49 (m, 4 H, H-5,
including s, 3.51, OCH3), 2.08–1.77 (2 m, 2 H,
H-3%a,b), 1.20 (d, 3 H, J5,6 6.2 Hz, H-6); 13C
NMR (D2O): l 177.27 (CO), 101.16 (C-1),
71.12 (C-5), 70.63 (C-3), 69.84 (C-2), 69.08
(C-2%), 57.93 (C-4%), 56.92 (OCH3), 53.00 (C-4),
36.05 (C-3%), 16.93 (C-6); CIMS: m/z 280
([M+1]+), 297 ([M+18]+). Anal. Calcd for
C11H21NO8·0.5 H2O: C, 44.43; H, 7.67; N, 4.71.
Found: C, 44.55; H, 7.58; N, 4.55.
1
ane), [h]D −109° (c 1, CHCl3); H NMR
(CDCl3): l 4.34 (d, 1 H, J1,2 1.0 Hz, H-1), 3.88
(dd, 1 H, J2,3 3.2 Hz, H-2), 3.53 (s, 3 H, OCH3),
3.29 (dd, 1 H, J3,4 10 Hz, H-3), 3.17 (m, 1 H,
H-5), 2.70 (t, 1 H, J 9.7 Hz, H-4), 1.32 (d, 3 H,
13
J5,6 6.2 Hz, H-6); C NMR (CDCl3): l 100.84
(C-1), 73.35 (C-3), 72.60 (C-5), 69.71 (C-2),
56.40 (OCH3), 54.12 (C-4), 17.28 (C-6); CIMS:
m/z 178 ([M+1]+), 195 ([M+18]+). Anal.
Calcd for C7H15NO4: C, 47.45; H, 8.53; N,
7.90. Found: C, 47.32; H, 8.46; N, 7.80.
Methyl 2,3-di-O-acetyl-4-(2,4-di-O-acetyl-
Methyl 4-azido-3-O-benzyl-2-O-methyl-4,6-
dideoxy-i-D-mannopyranoside
(7).—Iodo-
3-deoxy-
i- -mannopyranoside (28) and methyl 4-(3-de-
oxy- -glycero-tetronamido)-4,6-dideoxy-i-
mannopyranoside (6).—A mixture of amine 5
(356 mg, 2 mmol), 3-deoxy- -glycero-tetrono-
L
-glycero-tetronamido)-4,6-dideoxy-
methane (0.52 mL, 8.5 mmol) was added to a
stirred mixture of the b-glycoside 4 (1.66 g,
5.66 mmol) and powdered KOH (1 g, ꢀ20
mmol) in Me2SO (20 mL), and stirring was
continued for 1 h). TLC (2:1 hexane–acetone)
showed that the reaction was complete and
that one faster moving product was formed.
After filtration through a sintered glass funnel
(coarse porosity), water (100 mL) was added to
the filtrate, pH was adjusted to 7.5 by addition
of dilute AcOH, and the product was extracted
into CH2Cl2. The organic phase was dried,
concd, and the residue was passed through a
short column of silica gel to give the title
compound 7 (1.7 g, 98%) as an oil, [h]D +18°
(c 0.9, CHCl3). 1H NMR (CDCl3): l 4.74, 4.68
D
L
D-
L
lactone (490 mg, 4.8 mmol) and pyridine (0.2
mL) was heated at 110 °C (bath) overnight.
After cooling to rt, pyridine (2 mL) and Ac2O
(2 mL) was added, and the mixture was kept at
rt overnight. After concentration, chromatog-
raphy and crystallization gave the fully acetyl-
ated compound 28 (0.96 g, 77.5%), mp
139–139.5 °C (from EtOAc–2-Pr2O), [h]D
1
+30° (c 0.75, CHCl3). H NMR (CDCl3): l
6.01 (d, 1 H, J4,NH 9.3 Hz, NH), 5.43 (dd, 1 H,
J1,2 0.9, J2,3 3.3 Hz, H-2), 5.07 (dd, partially
overlapped, J2%,3a% 5.07, J2%,3b% 7.9 Hz, H-2%), 5.04
(dd, partially overlapped, J3,4 10.8 Hz, H-3),
4.48 (d, 1 H, H-1), 4.20–4.06 (m, 3 H, H-
4,4%a,b), 3.51 (s, partially overlapped, OCH3),
3.52–3.42 (m, partially overlapped, H-5),
2.22–2.03 (m, 14 H, H-3%a,b, including 4 s at
2.19, 2.16, 2.05, 2.04, 4 COCH3), 1.32 (d, 3 H,
J5,6 6.2 Hz, H-6); 13C NMR (CDCl3): l 171.07,
170.27, 169.66, 169.43, 162.53 (5 CO), 99.46
(C-1), 71.98 (C-5), 70.99 (C-2%), 70.31 (C-3),
68.37 (C-2), 59.77 (C-4%), 57.18 (OCH3), 51.52
(C-4), 30.53 (C-3%), 20.90, 20.82, 20.72, 20.67 (4
COCH3), 17.78 (C-6); CIMS: m/z 465 ([M+
18]+). Anal. Calcd for C19H29NO11: C, 51.00;
H, 6.53; N, 3.13. Found: C, 51.15; H, 6.69; N,
3.23.
2
(2 d, 1 H each, J 11.9 Hz, CH2Ph), 4.19 (d, 1
H, J1,2 0.7 Hz, H-1), 3.61 (s, partially over-
lapped, OCH3-2), 3.60 (bd, partially over-
lapped, H-2), 3.48 (s, partially overlapped,
OCH3-1), 3.47 (t, partially overlapped, J 9.4
Hz, H-4), 3.35 (dd, 1 H, J2,3 2.9, J3,4 9.9 Hz,
H-3), 3.14–3.05 (m, 1 H, H-5), 1.38 (d, 3 H, J5,6
6.3 Hz, H-6); 13C NMR (CDCl3): l 101.98
(C-1, JC-1,H-1 154.0 Hz), 80.52 (C-3), 76.65
(C-2), 71.75 (CH2Ph), 71.05 (C-5), 64.00 (C-4),
61.29 (OCH3-2), 57.03 (OCH3-1), 18.41 (C-6);
CIMS: m/z 325 ([M+18]+). Anal. Calcd for
C15H21N3O4: C, 58.62; H, 6.89; N, 13.67.
Found: C, 58.57; H, 6.84; N, 13.77.
Methyl 4-amino-4,6-dideoxy-2-O-methyl-i-
-mannopyranoside (8).—The 2-O-methyl
D

E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
13
TLC (6:1 toluene–acetone) showed that the
starting diol was essentially consumed and
that three products were formed. The phases
were separated, the aq soln was extracted with
CH2Cl2, and the combined solutions in
CH2Cl2 were washed with satd aq NaCl. The
organic phase was dried and concd, and the
residue was chromatographed.
derivative 7 (1.68 g) was treated with H2, as
described above for the preparation of 5, to
give the title compound 8 (0.89 g, 85%), mp
152.5–153 °C (from EtOH–hexane), [h]D
1
−124° (c 0.9, MeOH). H NMR (CDCl3–
CD3OD): l 4.35 (d, 1 H, J1,2 0.8 Hz, H-1),
3.62 (s, 3 H, OCH3-2), 3.53–3.51 (m, 4 H,
including s at 3.52 for OCH3-1, H-2), 3.27 (dd,
1 H, J2,3 3.5, J3,4 10.1 Hz, H-3), 3.18–3.09 (m,
1 H, H-5), 2.63 (t, 1 H, J 9.6 Hz, H-4), 1.32
The 2,3-di-O-methyl derivative 13 eluted
first was a colorless oil (0.7 g, 12.4%), [h]D
1
+125° (c 0.7, CHCl3); H NMR (CDCl3): l
13
(d, 3 H, J5,6 6.3 Hz, H-6); C NMR (CDCl3–
4.74 (d, 1 H, J1,2 1.8 Hz, H-1), 3.57 (dd, 1 H,
J2,3 3.0 Hz, H-2), 3.52, 3.50 (2 s, partially
overlapped, OCH3-2,3), 3.52–3.39 (m, 3 H,
H-3,5 including t at 3.42, H-4), 3.35 (s, 3 H,
OCH3-1), 1.31 (d, 3 H, J5,6 5.7 Hz, H-6). 13C
NMR (CDCl3): l 98.16 (C-1), 79.96 (C-3),
75.69 (C-2), 66.85 (C-5), 63.98 (C-4), 59.15,
57.24 (2 OCH3-2,3), 54.90 (OCH3-1), 18.38
(C-6). CIMS: m/z 249 ([M+18]+). Anal.
Calcd for C9H17N3O4: C, 46.74; H, 7.41; N,
18.17. Found: C, 46.68; H, 7.35; N, 18.17.
Eluted next was the 2-O-methyl derivative
12 (2.66 g, 50.2%), mp 57–57.5 °C (from 2-
CD3OD): l 102.31 (C-1), 79.58 (C-2), 73.29
(C-3), 72.70 (C-5), 61.48 (OCH3-2), 56.61
(OCH3-1), 54.76 (C-4), 17.29 (C-6); CIMS:
m/z 192 ([M+1]+), 209 ([M+18]+). Anal.
Calcd for C8H17NO4: C, 50.25; H, 8.96; N,
7.32. Found: C, 50.20; H, 8.95; N, 7.40.
Methyl 4-(3-deoxy-
4,6-dideoxy-2-O-methyl-i-
(9).—A mixture of amine 8 (640 mg, 3.35
mmol), 3-deoxy- -glycero-tetronolactone (684
L
-glycero-tetronamido)-
D-mannopyranoside
L
mg, 6.7 mmol) and pyridine (0.2 mL) was
stirred at 110 °C (bath) overnight. TLC (5:1
CH2Cl2 –MeOH) showed that all starting
amine was consumed. Elution from a small
column of silica gel gave 9 (702 mg, 75%),
which solidified on standing, mp 229–
229.5 °C (from EtOH), [h]D −108° (c 0.9,
1
Pr2O), [h]D +71° (c 0.8, CHCl3); H NMR
(CDCl3): l 4.76 (bd, 1 H, H-1), 3.82 (ddd,
partially overlapped, 1 H, J2,3 3.8, J3,4 10.1,
J3,OH 10.1 Hz, H-3), 3.53–3.43 (m, 5 H, H-5,
including s, 3.48 for OCH3-2 and dd, J1,2
ꢀ1.4 Hz for H-2), 3.36 (s, 3 H, OCH3-1), 3.19
(t, 1 H, H-4), 1.32 (d, 3 H, J5,6 6.0 Hz, H-6);
13C NMR (CDCl3): l 97.02 (C-1), 79.30 (C-2),
70.27 (C-3), 66.53 (C-4), 66.29 (C-5), 58.75
(OCH3-2), 54.93 (OCH3-1), 18.29 (C-6);
CIMS: m/z 235 ([M+18]+) Anal. Calcd for
C8H15N3O4: C, 44.23; H, 6.96; N, 19.34.
Found: C, 44.52; H, 7.07; N, 19.55.
1
MeOH); H NMR (CDCl3–CD3OD): l 4.38
(d, 1 H, J1,2 ꢀ0.6 Hz, H-1), 4.21 (dd, 1 H,
J2%,3a% 4.0, J2%,3b% 7.6 Hz, H-2%), 3.80–3.74 (m, 3
H, H-4,4%a,b), 3.62 (s, partially overlapped,
OCH3-2), 3.60 (dd, partially overlapped, J2,3
3.3, J3,4 10.3 Hz, H-3), 3.57 (dd, 1 H, H-2),
3.52 (s, 3 H, OCH3-1), 3.49–3.40 (m, 1 H,
H-5), 2.08–1.97, 1.91–1.78 (2 m, 1 H each,
H-3%a,b), 1.26 (d, 3 H, J5,6 6.3 Hz, H-6); 13C
NMR (CDCl3–CD3OD): l 101.86 (C-1),
79.56 (C-2), 70.88 (C-3), 70.51 (C-5), 69.38
(C-2%), 60.88 (OCH3-2), 58.01 (C-4%), 56.16
(OCH3-1), 53.26 (C-4), 35.94 (C-3%), 16.83 (C-
6); CIMS: m/z 294 ([M+1]+), 311 ([M+18]+).
Anal. Calcd for C11H23NO7: C, 49.14; H, 7.90;
N, 4.78. Found: C, 49.18; H, 8.02; N, 4.77.
Methyl 4-azido-2-O- (12), 3-O- (11) and 2,3-
Eluted last was the amorphous 3-O-methyl
derivative 11 (1.18 g, 22.3%), [h]D +156.5° (c
1
1.2, CHCl3); H NMR (CDCl3): l 4.73 (d, J1,2
1.5 Hz, H-1), 4.02 (dd, 1 H, J2,3 3.0 Hz, H-2),
3.56–3.44 (m, 5 H, H-3,5, including s at 3.50
for OCH3-2), 3.36 (s, partially overlapped,
OCH3-1), 3.34 (t, partially overlapped, J 9.7
Hz, H-4), 1.32 (d, 3 H, J5,6 6.3 Hz, H-6); 13C
NMR (CDCl3): l 100.90 (C-1), 80.55 (C-3),
66.52 (C-2), 66.35 (C-5), 63.69 (C-4), 57.22
(OCH3-3), 54.97 (OCH3-1), 18.32 (C-6);
CIMS: m/z 235 ([M+18]+). Found: C, 44.37;
H, 7.01; N, 19.13.
di-O-methyl-4,6-dideoxy-h- -mannopyranoside
D
(13).—A mixture of compound 10,11–13 (5 g,
23.6 mmol) Bu4NHSO4 (1.28 g), 20% NaOH
(120 mL) and MeI (5.9 mL, 94.8 mmol) in
CH2Cl2 (120 mL) was stirred at rt overnight.
Methyl 4-(3-deoxy-
L
-glycero-tetronamido)-
-mannopyranoside
4,6-dideoxy-3-O-methyl-h-
D

14
E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
dd at 3.67, J3,4 10.6 Hz, H-3), 2.25 (t, 2 H, J
7.6 Hz, H-2%), 1.22 (d, 3 H, J5,6 6.2 Hz, H-6),
1.15 (t, 3 H, H-3%); 13C NMR (4:1 CDCl3–
CD3OD): l 100.78 (C-1), 69.52 (C-2), 69.30
(C-3), 54.43 (OCH3), 52.93 (C-4), 29.21 (C-2%),
17.35 (C-6), 9.55 (C-3%); CIMS: m/z 234
([M+1]+), 252 ([M+18]+). Anal. Calcd for
C10H19NO5: C, 51.49; H, 8.21; N, 6.00. Found:
C, 51.34; H, 8.14; N, 5.92.
(15).—Compound 11 (1.19 g) was treated
with H2 as described for the preparation of 8.
The crude product was purified by chromatog-
raphy to give amorphous, hygroscopic methyl
4 - amino - 4,6 - dideoxy - 3 - O - methyl - a - -
D
mannopyranoside (14) (725 mg, 75%). 1H
NMR (CDCl3–CD3OD): l 4.70 (d, 1 H, J1,2
1.5 Hz, H-1), 4.01 (dd, 1 H, J2,3 3.0 Hz, H-2),
3.61–3.49 (m, 1 H, H-5), 3.43 (s, 3 H, OCH3-
3), 3.37 (s, 3 H, OCH3-1), 3.24 (dd, 1 H, J3,4
9.9 Hz, H-3), 2.83 (t, 1 H, J 9.8 Hz, H-4), 1.28
Methyl 4,6-dideoxy-2-O-methyl-4-propan-
amido-h- -mannopyranoside (23).—A mixture
D
13
of azide 12 (740 mg, 3.4 mmol), ESCAT 103
(0.7 g) and propanoic anhydride (0.87 mL, 6.8
mmol) was treated with H2 as described for
the preparation of 18 to give the title com-
pound 23 (660 mg, 78%), mp 188.5–189 °C
(d, 3 H, J5,6 6.4 Hz, H-6); C NMR (CDCl3–
CD3OD): l 100.89 (C-1), 80.42 (C-3), 68.35
(C-5), 64.68 (C-2), 55.96 (OCH3-3), 54.47
(OCH3-1), 52.65 (C-4), 17.41 (C-6), CIMS:
m/z 192 ([M+1]+), 209 ([M+18]+).
1
(from EtOH). [h]D +65° (c 0.9, CHCl3). H
Amine 14 (380 mg, 2 mmol) was treated
NMR (CDCl3): l 4.37 (d, 1 H, J1,2 1.5 Hz,
H-1), 3.86 (t, 1 H, J 10.3 Hz, H-4), 3.67 (dd,
1 H, J2,3 3.4, J3,4 10.7 Hz, H-3), 3.59 (m, 1 H,
H-5), 3.50 (s, 3 H, OCH3-2), 3.42 (dd, 1 H,
H-2), 3.37 (s, 3 H, OCH3-1), 2.25 (q, 1 H, J2%,3%
7.6 Hz, H-2%), 1.20 (d, 3 H, J5,6 6.2 Hz, H-6),
1.15 (t, 3 H, H-3%). Before deuterium ex-
change, the NH and H-3 signals appear at l
5.46 (s) and 3.69 (dt). 13C NMR (CDCl3): l
97.44 (C-1), 79.28 (C-2), 69.81 (C-3), 67.35
(C-5), 58.87 (OCH3-2), 54.91 (OCH3-1), 54.10
(C-4), 29.82 (C-2%), 17.93 (C-6), 9.82 (C-3%).
CIMS: m/z 248 ([M+1]+), 265 ([M+18]+).
Anal. Calcd for C11H21NO5: C, 53.43; H, 8.56;
N, 5.66. Found: C, 53.31; H, 8.45; N, 5.52.
with 3-deoxy- -glycero-tetronolactone (858
L
mg, 8.4 mmol) as described for the prepara-
tion of 9. Chromatography and crystallization
from acetone gave 15 (381 mg, 65%), mp
137–137.5 °C (from acetone), [h]D +43° (c
1
0.8, MeOH). H NMR (CDCl3 –CD3OD): l
4.72 (d, 1 H, J1,2 1.7 Hz, H-1), 4.25 (dd, 1 H,
J2%,3%a 3.9, J2%,3%b 8.2 Hz, H-2%), 4.05 (dd, 1 H,
J2,3 2.9 Hz, H-2), 3.96 (t, 1 H, J 10.3 Hz, H-4),
3.80–3.69 (m, 3 H, H-5,4%a,b), 3.47 (dd, 1 H,
J3,4 10.4 Hz, H-3), 3.39 (s, 3 H, OCH3-1), 3.38
(s, 3 H, OCH3-3), 2.09–1.98, 1.88–1.76 (2 m,
1 H each, H-3%a,b), 1.23 (d, 3 H, J5,6 6.2 Hz,
H-6); 13C NMR (CDCl3–CD3OD): l 100.52
(C-1), 78.16 (C-3), 69.87 (C-2%), 66.81 (C-5),
65.51 (C-2), 58.66 (C-4%), 56.35 (OCH3-3),
54.51 (OCH3-1), 51.14 (C-4), 36.09 (C-3%),
17.25 (C-6); CIMS: m/z 294 ([M+1]+), 311
([M+18]+). Anal. Calcd for C12H23NO7: C,
49.14; H, 7.90; N, 4.78. Found: C, 49.21; H,
7.78; N, 4.84.
Methyl 4,6-dideoxy-4-(
L
-glycero-2-hydroxy-
-mannopyranoside (19) and
2,3-di-O-acetyl-4,6-dideoxy-4-(2-O-
-glycero-propanamido)-h- -manno-
propanamido)-h-
D
methyl
acetyl-
L
D
pyranoside (29).—A mixture of amine 1612
(558 mg, 3 mmol),
L
-lactic acid (270 mg, 9
mmol), EDAC (1.72 g, 9 mmol) and HOBT
(1.28 g, 9 mmol) in 1:2 MeCN–CH2Cl2 (15
mL) was stirred at rt overnight. TLC (2:1
CH2Cl2–MeOH) then showed that all amine
had been consumed. After concentration, the
residue was chromatographed to give 19 that
still contained some UV positive material.
Acetylation, as described below for the prepa-
ration of 28, gave 29 (0.97 g, 82.7%), mp
131.5–132 °C (from EtOAc–2-Pr2O), [h]D
Methyl
4,6-dideoxy-4-propanamido-h- -
D
mannopyranoside (18).—A mixture of azide
1011–13 (1.29 g, 5.9 mmol), propanoic anhy-
dride (1.5 mL, 11 mmol) and ESCAT 103 (0.5
g) was stirred in a H2 atmosphere for 2 h.
After concentration, chromatography and
crystallization gave the title compound 18
(1.28 g, 92%), mp 171–171.5 °C (from EtOH),
1
[h]D +82° (c 0.8, MeOH). H NMR (4:1
1
CDCl3–CD3OD): l 4.66 (d, 1 H, J1,2 1.6 Hz,
H-1), 3.87 (t, partially overlapped, J 10.2,
H-4), 3.82 (dd, partially overlapped, J2,3 3.4
Hz, H-2), 3.70–3.58 (m, 2 H, H-5, including
+63° (c 0.8, CHCl3). H NMR (CDCl3): l
6.12 (d, 1 H, J4,NH 9.4 Hz, NH), 5.27 (dd, 1 H,
J2,3 3.4, J3,4 11.1 Hz, H-3), 5.14 (dd, 1 H, J1,2
1.8 Hz, H-2), 5.07 (q, 1 H, J2%,3% 6.9 Hz, H-2%),

E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
15
H, OCH3-2), 3.44 (dd, 1 H, H-2), 3.38 (s, 3 H,
OCH3-1), 1.40 (d, 3 H, H-3%), 1.21 (s, 3 H, J5,6
6.2 Hz, H-6); 13C NMR (CDCl3–CD3OD): l
97.57 (C-1), 79.29 (C-2), 68.91 (C-3), 67.95
(C-2%), 66.88 (C-5), 58.72 (OCH3-2), 54.66
(OCH3-1), 53.48 (C-4), 20.16 (C-3%), 17.41 (C-
6). CIMS: m/z 264 ([M+1]+), 281 ([M+18]+).
Anal. Calcd for C11H21NO6: C, 50.18; H, 8.04;
N, 5.32. Found: C, 50.28; H, 8.01; N, 5.24.
Methyl 4,6-dideoxy-4-(hydroxyacetamido)-
4.66 (d, 1 H, H-1), 4.22 (2 t, partially over-
lapped, H-4), 3.75–3.65 (m, 1 H, H-5), 3.39 (s,
3 H, OCH3), 2.17, 2.15, 2.04 (3 s, 3 H each, 3
COCH3), 1.42 (d, 3 H, H-3%), 1.25 (d, 3 H, J5,6
6.2 Hz, H-6); 13C NMR (CDCl3): l 171.03,
170.72, 170.01, 169.48 (4 CO), 98.30 (C-1),
70.44 (C-2%), 69.17 (C-2), 68.36 (C-3), 67.95
(C-5), 55.02 (OCH3), 51.21 (C-4), 20.92, 20.86,
20.70 (3 COCH3), 17.67 (C-6), 17.55 (C-3%);
CIMS: m/z 250 ([M+1]+), 267 ([M+18]+).
Anal. Calcd for C16H25NO9: C, 51.19; H, 6.71;
N, 3.73. Found: C, 51.40; H, 6.74; N, 3.76.
Compound 29 was deacetylated (Zemple´n)
to give the crystalline hemihydrate of 19 in
virtually theoretical yield, mp 137.5–138 °C
(from acetone–hexane), [h]D +66° (c 0.6,
h-
D
-mannopyranoside (20).—Methyl 4-amino-
4,6-dideoxy-a-
D
-mannopyranoside (16) (224
mg, 1.26 mmol)12 was treated with glycolic
acid (123 mg, 1.62 mmol) as described below
for the corresponding 2-O-methyl derivative,
to give 20 (240 mg, 81%), mp 145.5–146 °C
1
MeOH). H NMR (CDCl3–CD3OD): l 7.25
1
(from acetone), [h]D +67° (c 0.8, H2O). H
(d, 1 H, J4,NH 9.4 Hz, NH), 4.68 (d, 1 H, J1,2
1.6 Hz, H-1), 4.17 (q, 1 H, J2%,3% 6.8 Hz, H-3%),
3.88–3.81 (m, 2 H, H-2,4), 3.78–3.67 (m, 2 H,
H-3,5), 3.37 (s, 3 H, OCH3), 1.40 (d, 3 H,
H-3%), 1.22 (d, 3 H, J5,6 6.2 Hz, H-6); 13C
NMR (CDCl3): l 100.82 (C-1), 69.58 (C-2),
69.14 (C-3), 67.86 (C-2%), 66.78 (C-5), 54.54
(OCH3), 53.06 (C-4), 20.17 (C-3%), 17.36 (C-6).
Anal. Calcd for C10H19NO6·0.5 H2O: C, 46.50;
H, 7.80; N, 5.42. Found: C, 46.68; H, 7.73; N,
5.40.
NMR (CDCl3–CD3OD): l 4.68 (d, 1 H, J1,2
1.7 Hz, H-1), 4.07, 4.00 (2 d, 2 H, J 16.3 Hz,
2
H-2%a,b), 3.92 (t, 1 H, J 10.3 Hz, H-4), 3.85
(dd, 2 H, J2,3 3.3 Hz, H-2), 3.78–3.67 (m, 2 H,
including dd for H-3 at 3.74, J3,4 10.5 Hz,
H-5), 3.38 (s, 3 H, OCH3), 1.23 (d, 3 H, J5,6
6.2 Hz, H-6); 13C NMR (CDCl3–CD3OD): l
100.77 (C-1), 69.45 (C-2), 49.13 (C-3), 66.73
(C-5), 61.38 (C-2%), 54.47 (OCH3), 52.73 (C-4),
17.30 (C-6); CIMS: m/z 236 ([M+1]+), 253
([M+18]+). Anal. Calcd for C9H17NO6: C,
45.95; H, 7.28; N, 5.95. Found: C, 46.01; H,
7.26; N, 6.02.
Methyl 4,6-dideoxy-4-( -glycero-2-hydroxy-
L
propanamido)-2-O-methyl-h- -mannopyran-
D
oside (24).—A mixture of amine 1718 (pre-
pared from azide 12 as described below, 191
Compound 20 was further characterized as
methyl 2,3-di-O-acetyl-4,6-dideoxy-4-(acetoxy-
mg, 1 mmol),
L
-lactic acid (108 mg, 1.2
acetamido)-a- -mannopyranoside (30), mp
D
mmol), EDAC (574 mg, 3 mmol) and 1-hy-
droxybenzotriazole (406 mg, 3 mmol) in
CH2Cl2 (10 mL) was stirred at rt overnight.
TLC (6:1 CH2Cl2–MeOH) then showed that
all amine was consumed and that one major,
faster moving product was formed. The mix-
ture was washed successively with 2 M HCl,
aq NaHCO3 and NaCl, dried, and concd, and
the residue was chromatographed to give the
pure title compound 24 (205 mg, 78%), mp
165.5–166 °C (from EtOH–hexane), [h]D
+50°(c1,CHCl3).1HNMR(CDCl3–CD3OD):
l 7.13 (d, B1 H, due to incomplete H“D
exchange, J1,2 9.3 Hz, NH), 4.77 (d, 1 H, J1,2
1.4 Hz, H-1), 4.15 (q, 1 H, J2%,3% 6.9 Hz, H-2%),
3.82 (t, 1 H, J 10.3, H-4), 3.75 (dd, partially
overlapped, J2,3 3.1, J3,4 10.3 Hz, H-3), 3.75–
3.63 (m, partially overlapped, H-5), 3.51 (s, 3
158–161 °C (from EtOH), [h]D +77° (c 0.9,
1
CHCl3). H NMR (CDCl3): l 6.12 (d, 1 H,
J4,NH 9.3 Hz, NH), 5.27 (dd, 1 H, J3,4 11.1, J2,3
3.4 Hz, H-3), 5.13 (dd, 1 H, J1,2 1.8 Hz, H-2),
2
4.67 (d, 1 H, H-1), 4.68 (d, 1 H, J 15.5 Hz,
H-2%a), 4.40 (d, 1 H, H-2%b), 4.25 (q, 1 H, J
10.1 Hz, H-4), 3.72 (m, 1 H, H-5), 3.39 (s, 3
H, OCH3), 2.19, 2.16, 2.01 (3 s, 3 H each, 3
COCH3), 1.28 (d, 3 H, J5,6 6.3 Hz, H-6). 13C
NMR (CDCl3): l 171.9, 170.57, 170.05,
167.96 (4 CO), 98.57 (C-1), 69.46 (C-2), 68.40
(C-3), 68.34 (C-5), 62.75 (C-2%), 55.22 (OCH3),
51.64 (C-4), 21.01, 20.80, 20.68 (3 CH3CO),
17.89 (C-6); CIMS: m/z 379 ([M+18]+).
Anal. Calcd for C15H23NO9: C, 49.86; H, 6.42;
N, 3.88; O, 39.85. Found: C, 49.94; H, 6.41;
N, 3.88.

16
E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
C, 48.19; H, 7.68; N, 5.62; O, 38.51. Found:
Methyl 4,6-dideoxy-4-(hydroxyacetamido)-
2-O-methyl-h- -mannopyranoside (25).—
C, 48.29; H, 7.71; N, 5.66.
D
Methyl 2,3-di-O-acetyl-4,6-dideoxy-4-(2,3-
Azide 12 was treated with H2 as described for
the preparation of 5 to give methyl 4-amino-
di-O-acetyl-
amido)-h- -mannopyranoside (32).—To
suspension of -glyceric acid hemicalcium salt
D
- (34) and
L
-glycero-propan-
D
a
4,6-dideoxy-2-O-methyl-a- -mannopyranoside
D
L
(17), which was identical (NMR) with the
previously described, independently synthe-
sized compound.18 To a soln of the foregoing
amine 17 (450 mg, 2.35 mmol) and glycolic
acid (358 mg, 4.7 mmol) in MeCN (25 mL)
was added HATU (1.79 g, 4.7 mmol), and
diisopropylethylamine (0.85 mL, 4.7 mmol),
and the mixture was stirred for 1 h at rt. TLC
(5:1 CHCl3–MeOH) then showed that all
starting amine had been consumed. After con-
centration and chromatography the material
that was obtained, which still contained some
impurities, was acetylated to give methyl 3-O-
acetyl - 4,6 - dideoxy - 4 - (acetoxyacetamido) - 2-
monohydrate (2.0 g, 13.97 mmol) in pyridine
(100 mL) was added Ac2O (15 mL), and the
mixture was stirred at rt for 24 h. Water (75
mL) was added at 0 °C to the resulting clear
solution, and the mixture was stirred for 2 h
to decompose the excess Ac2O. The soln was
concd and co-evaporated with toluene (3×
150 mL). A suspension of the residue in
CHCl3 (400 mL) was washed with 10% HCl
satd with NaCl (80 mL), and the aq layer was
extracted with CHCl3 (3×100 mL). The or-
ganic layers were combined, washed with
brine (100 mL), dried, and concd to give
O-methyl-a-
D
-mannopyranoside (31) (516 mg,
2,3-di-O-acetyl- -glyceric acid (38) (2.52 g,
L
66%), mp 143.5–144.5 °C (from EtOAc), [h]D
97%), which was sufficiently pure for the next
1
1
+105° (c 0.4, CHCl3); H NMR (CDCl3): l
step. H NMR (CDCl3): l 9.11 (bs, 1 H,
6.04 (d, 1 H, J4,NH 9.2 Hz, NH), 5.20 (dd, 1 H,
J2,3 3.2, J3,4 11.3 Hz, H-3), 4.75 (d, 1 H, J1,2
COOH), 5.37 (dd, 1 H, J2,3a 3.2, J2,3b 5.1 Hz,
2
H-2), 4.54 (dd, 1 H, J 12.1 Hz, H-3a), 4.45
2
1.4 Hz, H-1), 4.67, 4.38 (2 s, 1 H each, J 15.5
(dd, 1 H, H-3b), 2.19, 2.11 (2 s, 3 H each, 2
COCH3); 13C NMR (CDCl3): l 171.85,
170.97, 170.42 (3 CO), 69.92 (C-2), 62.49 (C-
3), 20.48, 20.30 (2 CH3); CIMS: m/z 208
([M+18]+).
Hz, H-2%a,b), 4.28 (q, 1 H, J 9.9 Hz, H-4),
3.70–3.60 (m, 1 H, H-5), 3.51 (s, 3 H, OCH3-
2), 3.49 (dd, 1 H, H-2), 3.38 (s, 3 H, OCH3-1),
2.19, 2.08 (2 s, 3 H each, 2 COCH3), 1.26 (d,
3 H, J5,6 6.3 Hz, H-6); 13C NMR (CDCl3): l
98.58 (C-1), 77.85 (C-2), 70.89 (C-3), 68.42
(C-5), 62.57 (C-2%), 59.57 (OCH3-2), 54.92
(OCH3-1), 51.57 (C-4), 20.94, 20.68 (2
COCH3), 17.92 (C-6); CIMS: m/z 334 ([M+
1]+), 351 ([M+18]+). Anal. Calcd for
C14H23NO8: C, 50.44; H, 6.95; N, 4.20; O,
38.40. Found: C, 50.45; H, 7.01; N, 4.13.
Deacetylation of 31 gave the title substance
25, [h]D +40° (c 0.8, H2O). 1H NMR
(CDCl3): l 7.03 (d, 1 H, J4,NH 9.7 Hz, NH),
4.78 (d, 1 H, J1,2 1.7 Hz, H-1), 4.06 (dd, 2 H,
2J 16.4 Hz, H-2%a,b), 3.90 (dd, 1 H, J3,4 10.1,
J4,5 10.1 Hz, H-4), 3.78 (bdd, 1 H, J2,3 ꢀ2.8
Hz, H-3), 3.67 (m, 1 H, H-5), 3.50 (s, 3 H,
OCH3-2), 3.45 (dd, 1 H, H-2), 3.38 (s, 3 H,
OCH3-1), 1.24 (d, 3 H, H-6); 13C NMR
(CDCl3): l 174.13 (CO), 97.39 (C-1), 79.27
(C-2), 69.29 (C-3), 66.91 (C-5), 62.21 (C-2%),
58.79 (OCH3-2), 54.90 (OCH3-1), 53.58 (C-4),
17.82 (C-6); CIMS: m/z 250 ([M+1]+) and
267 ([M+18]+). Anal. Calcd for C10H19NO6:
To a soln of the foregoing acid 38 (1.59 g,
8.38 mmol) and amine 16 (1.41 g, 6.44 mmol)
in MeCN (80 mL) was added HATU (3.18 g,
8.38 mmol), and the mixture was stirred for 4
h at rt. N,N-Diisopropylamine (DIPEA, 1.43
mL, 8.38 mmol) was added dropwise to com-
plete the conversion, and after 30 min when
TLC (5:1 CH2Cl2–Et2O) showed complete dis-
appearance of the starting amine, 10 mL of
pyridine and 10 mL of Ac2O was added, and
stirring was continued overnight. The mixture
was concd, and a soln of the residue in
CH2Cl2 was washed successively with 1 N HCl
and aq NaHCO3. After concentration, chro-
matography gave first the -glycero isomer 34
D
(0.511 g, 18.3%), mp 98–100 °C (from Et2O),
1
[h]D +77° (c 1, CHCl3). H NMR (CDCl3): l
6.31 (d, 1 H, J4,NH 9.6 Hz, NH), 5.34 (dd, 1 H,
J2,3a 2.8, J2,3b 4.3 Hz, H-2%), 5.28 (dd, 1 H, J2,3
3.2, J3,4 11.0 Hz, H-3), 5.12 (dd, 1 H, J1,2 1.8
2
Hz, H-2), 4.67 (d, 1 H, H-1), 4.45 (dd, 1 H, J
12.3 Hz, H-3%a), 4.38 (dd, 1 H, H-3%b), 4.23 (q,

E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
17
mixture of amine 17 (300 mg, 1.57 mmol),
acid 38 (650 mg, 3.42 mmol) and HATU (775
mg, 2.04 mmol) in MeCN (20 mL) was stirred
at rt until TLC (3:1 CH2Cl2–MeOH) showed
that all starting amine was consumed (ꢀ3 h).
Pyridine (4 mL) and Ac2O (4 mL) were added,
and stirring was continued overnight. After
concentration, a soln of the residue in CH2Cl2
was washed successively with 1 N HCl and aq
NaHCO3. Chromatography of the material in
1 H, J 10.0 Hz, H-4), 3.74 (m, 1 H, H-5), 3.39
(s, 3 H, OCH3), 2.21, 2.17, 2.07, 1.99 (4 s, 3 H
each, 4 COCH3), 1.29 (d, 3 H, J5,6 6.1 Hz,
13
H-6); C NMR (CDCl3): l 98.25 (C-1), 71.39
(C-2%), 69.25 (C-2), 68.16 (C-5), 67.86 (C-3),
63.38 (C-3%), 54.99 (OCH3), 51.70 (C-4), 20.81,
20.63, 20.54, 20.50 (4 COCH3), 17.46 (C-6);
CIMS: m/z 432 ([M−1]−), 451 ([M+18]+).
Anal. Calcd for C18H27NO11: C, 49.88; H,
6.28; N, 3.23. Found: C, 49.81; H, 6.37; N,
3.13.
the organic phase gave first the
D-glycero iso-
Eluted next was a mixture of the two
diastereoisomers (0.81 g, 29%).
mer 35 (0.16 g, 25%), mp 170–171 °C (from
EtOH–Et2O), [h]D +105° (c 0.75, CHCl3). 1H
NMR (CDCl3): l 6.20 (d, 1 H, J4,NH 11.0 Hz,
NH), 5.36 (dd, 1 H, J2%,3%a 2.8, J2%,3%b 4.3 Hz,
H-2%), 5.21 (dd, 1 H, J2,3 3.0, J3,4 11.3 Hz,
H-3), 4.75 (d, 1 H, J1,2 1.7 Hz, H-1), 4.45 (dd,
Eluted last was the
L
-glycero isomer 32
(1.18 g, 42.3%, total yield of N-acylation,
1
89.7%), [h]D +43° (c 0.8, CHCl3); H NMR
(CDCl3): l 6.27 (d, 1 H, J4,NH 9.6 Hz, NH),
5.27 (dd, partially overlapped, J2,3 3.4, J3,4
10.9 Hz, H-3), 5.25 (dd, partially overlapped,
J2,3a 3.7, J2,3b 5.6 Hz, H-2%), 5.15 (dd, 1 H, J1,2
1.8 Hz, H-2), 4.66 (d, 1 H, H-1), 4.53 (dd, 1
2
1 H, J2%,3a 3.0, J 12.1 Hz, H-3%a), 4.38 (dd,
J2%,3b 4.2 Hz, H-3%b), 4.28 (q, 1 H, J 10.4 Hz,
H-4), 3.66 (m, 1 H, H-5), 3.52 (d, 3 H, OCH3-
2), 3.47 (dd, 1 H, H-2), 3.39 (s, 3 H, OCH3-1),
2.22, 2.06, 2.05 (3 s, 3 H each, 3 COCH3), 1.27
2
H, J 12.0 Hz, H-3%a), 4.31 (dd, partially over-
2
13
lapped, 1 H, J 12.0 Hz, H-3%b), 4.22 (q,
(d, 3 H, J5,6 6.3 Hz, H-6); C NMR (CDCl3):
partially overlapped, J 10.3 Hz, H-4), 3.72 (m,
l 172.21, 170.19, 169.08 (4 CO), 98.67 (C-1),
77.95 (C-2), 71.43 (C-2%), 70.65 (C-3), 68.63
(C-5), 63.60 (C-3%), 59.59 (OCH3-2), 54.95
(OCH3-1), 51.87 (C-4), 20.87, 20.82, 20.64 (3
COCH3), 17.66 (C-6); CIMS: m/z 404 ([M+
1]+), 423 ([M+18]+). Anal. Calcd for
C17H27NO10: C, 50.37; H, 6.71; N, 3.46.
Found: C, 50.58; H, 6.71; N, 3.50.
1 H, H-5), 2.18, 2.17, 2.07, 2.05 (4 s, 3 H each,
13
4 COCH3); C NMR (CDCl3): l 98.29 (C-1),
71.56 (C-2%), 69.06 (C-2), 68.25 (C-3), 67.73
(C-5), 62.58 (C-3%), 55.01 (OCH3), 51.42 (C-4),
20.85, 20.67, 20.56, 20.52 (4 COCH3), 17.65
(C-6); CIMS: m/z 432 ([M−1]+), 451 ([M+
18]+). Found: C, 49.75, H, 6.35, N, 3.16.
Methyl
glycero - propanamido) - h -
4,6-dideoxy-4-(2,3-dihydroxy-
L
-
Eluted next was a mixture of the two
diastereoisomers (0.12 g, 19%).
D
- mannopyranoside
(22).—Deacetylation of compound 32 (1.03 g,
Zemple´n) gave, after chromatography, depro-
tected substance 22 (531 mg, 84%), mp 155–
156°C (from EtOH–EtOAc), [h]D +41° (c
0.8, H2O). 1H NMR (D2O): l 4.72 (d, 1 H, J1,2
1.4 Hz, H-1), 4.24 (dd, 1 H, J2,3%a 4.0, J2,3%b 4.9
Hz, H-3%), 3.93–3.75 (m, 6 H, H-2,3,4,5,3%a,b),
3.37 (s, 3 H, OCH3), 1.18 (d, 3 H, J5,6 5.8 Hz,
Eluted last was the
L
-glycero isomer 33
(0.215 g, 34%, total yield of N-acylation,
78%), mp 110–112 °C (from Et2O–hexane),
1
[h]D +66° (c 0.8, CHCl3). H NMR (CDCl3):
l 6.13 (d, 1 H, J4,NH 9.4 Hz, NH), 5.26 (dd, 1
H, J2%,3%a 3.8, J2%,3%b 5.3 Hz, H-2%), 5.21 (dd, 1 H,
J2,3 2.8, J3,4 11.0 Hz), 4.74 (d, 1 H, J1,2 1.9 Hz,
2
H-1), 4.53 (dd, 1 H, J 12.0 Hz, H-3%a), 4.30
13
H-6); C NMR (D2O): l 175.14 (CO), 100.99
(dd, partially overlapped, H-3%b), 4.25 (q, par-
tially overlapped, H-4), 3.66 (m, 1 H, H-5),
3.51 (m, 4 H, H-2, OCH3-2), 3.38 (s, 3 H,
OCH3-1), 2.18, 2.13, 2.07 (3 s, 3 H each, 3
COCH3), 1.23 (d, 3 H, J5,6 6.3 Hz, H-6); 13C
NMR (CDCl3): l 171.59, 170.42, 169.49,
167.00 (4 CO), 98.52 (C-1), 77.70 (C-2), 71.56
(C-2%), 70.81 (C-3), 67.98 (C-5), 62.67 (C-3%),
59.54 (OCH3-2), 54.91 (OCH3-1), 51.64 (C-4),
17.76 (C-6); CIMS: m/z 404 ([M+1]+), 423
(C-1), 72.61 (C-2%), 69.23 (C-2), 67.90 (C-3),
67.34 (C-5), 63.41 (C-3%), 54.85 (OCH3), 53.12
(C-4), 16.89 (C-6); CIMS: m/z 266 ([M+1]+),
283 ([M+18]+). Anal. Calcd for C10H19NO7:
C, 45.28; H, 7.22; N, 5.28. Found: C, 45.46;
H, 7.28; N, 5.33.
Methyl 3-O-acetyl-4,6-dideoxy-4-(2,3-di-O-
acetyl-
D
- (35) and
L
-glycero-propanamido)-2-
O-methyl-h-
D
-mannopyranoside (33).—To a

18
E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
([M+18]+). Found: C, 50.34; H, 6.72; N,
(C-4), 37.0 (C-2%), 17.9 (C-6); CIMS: m/z 340
([M+1]+) and 357 ([M+18]+). Anal. Calcd
for C17H25NO6: C, 60.16; H, 7.42; N, 4.13.
Found: C, 60.21; H, 7.45; N, 4.13.
3.49.
Methyl 4,6-dideoxy-4-(2,3-dihydroxy-
L
-gly-
cero-propanamido)-2-O-methyl-h-
D
-manno-
Methyl
4,6-dideoxy-4-(3-hydroxypropan-
-mannopyranoside (21).—A soln of
pyranoside (27).—Deacetylation of 33 (Zem-
ple´n) gave amorphous, hygroscopic 27 in vir-
tually theoretical yield, [h]D +18° (c 0.7,
amido)-h-
D
compound 36 (315 mg) in EtOH (10 mL) was
treated with H2 at atmospheric pressure and rt
in the presence of 5% Pd–C catalyst (ꢀ100
mg) to give 21 in virtually theoretical yield,
mp 134–136 °C (from acetone), [h]D +67° (c
1, H2O). 1H NMR (D2O): 4.70 (d, 1 H, J1,2 1.7
Hz, H-1), 3.89 (dd, 1 H, J2,3 2.9 Hz, H-2),
3.85–3.66 (m, 5 H, H-3,4,5,3%a,b), 3.66 (s, 3 H,
OCH3), 2.48 (t, 2 H, J 6.0 Hz, H-2%a,b), 1.17
1
H2O). H NMR (CDCl3): l 4.91 (d, 1 H, J1,2
1.7 Hz, H-1), 4.26 (dd, 1 H, J2%,3a 4.0, J2%,3b 5.2
Hz, H-2%), 3.97 (bdd, 1 H, J2,3 3.4, J3,4 10.5 Hz,
H-3), 3.86–3.77 (m, 4 H, H-4,5,3%a,b), 3.59
(dd, 1 H, H-2), 3.50 (s, 3 H, OCH3-2), 3.42 (s,
3 H, OCH3-1), 1.19 (d, 3 H, J5,6 5.8 Hz, H-6);
13C NMR (CDCl3): l 175.3 (CO), 97.82 (C-1),
79.07 (C-2), 72.56 (C-2%), 67.65 (C-3), 67.20
(C-5), 63.40 (C-3%), 58.92 (OCH3-2), 54.92
(OCH3-1), 53.54 (C-4), 16.87 (C-6). CIMS:
m/z 280 ([M+1]+), 297 ([M+18]+).
13
(d, 3 H, J5,6 6.0 Hz, H-6); C NMR (D2O): l
174.9 (CO), 100.95 (C-1), 69.15 (C-2), 68.34
(C-3), 67.54 (C-5), 57.99 (C-3%), 54.86 (OCH3),
53.02 (C-4), 38.90 (C-2%), 16.93 (C-6); CIMS:
m/z 250 ([M+1]+), 267 ([M+18]+). Anal.
Calcd for C10H19NO6: C, 48.19; H, 7.68; N,
5.62. Found: C, 48.39; H, 7.77; N, 5.42.
3-Benzyloxypropionic acid (39).—This com-
pound was prepared from b-propiolactone as
described by Li et al.,22 except that the
amount of HCl added to the aq phase before
extraction with CH2Cl2 was adjusted to make
Methyl 4,6-dideoxy-4-(3-benzyloxypropan-
1
it strongly acidic. H NMR data agreed with
amido)-2-O-methyl-h- -mannopyranoside (37).
D
those reported. 13C NMR (CDCl3): l 177.5
(CO), 137.7 (1 C, Ar), 128.3 (2 C, Ar), 127.66
(3 C, Ar), 72.9 (CH2Ph), 65.0 (C-3), 34.6
(C-2).
—To a soln of compound 17 (192 mg, 1
mmol) and 3-benzyloxypropionic acid (39)
(216 mg, 1.2 mmol) in MeCN (10 mL) was
added HATU (456 mg, 1.2 mmol), and the
mixture was stirred at rt for 30 min. After
addition of DIPEA (0.22 mL, 1.2 mmol), stir-
ring was continued and, after 2 h, the mixture
was processed as described in the preparation
of 33. Chromatography (neat AcOEt) gave
compound 37 (289 mg, 82%), mp 89–90 °C
(from acetone–Et2O), [h]D +38° (c 0.5,
Methyl 4,6-dideoxy-4-(3-benzyloxypropan-
amido)-h- -mannopyranoside (36).—HATU
D
(644 mg, 1.7 mmol), followed by DIPEA (0.3
mL, 1.7 mmol), was added to a soln of amine
1611–13 (250 mg, 1.41 mmol) and acid 39 (520
mg, 2.8 mmol) in MeCN (15 mL). The mix-
ture was stirred for 1 h at rt, when TLC (2:1
CH2Cl2 –MeOH) showed that all starting
amine was consumed. After concentration, a
soln of the residue in dichloromethane was
processed as described above for the prepara-
tion of 33, and chromatography gave 36 (434
mg, 90%), mp 122–124 °C (from EtOAc), [h]D
1
CHCl3). H NMR (CDCl3): l 6.47 (d, 1 H,
JNH,4 9.4 Hz, NH), 4.75 (d, 1 H, J1,2 1.5 Hz,
H-1), 4.51 (2 d, 2 H, J 11.8 Hz, CH2Ph), 3.87
(m, 1 H, H-4), 3.73 (t, 2 H, J 5.7 Hz, H-3%a,b),
3.66 (dd, 1 H, J2,3 3.4, J3,4 10.5 Hz, H-3), 3.49
(m, 1 H, H-5), 3.45 (s, 3 H, OCH3-2), 3.42
(dd, 1 H, H-2), 3.34 (s, 3 H, OCH3-1), 2.51
(dd, 2 H, J2,3%a 6.9, J2,3%b 5.4 Hz, H-2%), 1.18 (d,
3 H, J5,6 6.2 Hz, H-6); 13C NMR (CDCl3): l
172.9 (CO), 97.2 (C-1), 79.2 (C-2), 73.0
(CH2Ph), 69.3 (C-3), 66.9 (C-5), 66.2 (C-3%),
58.5 (OCH3-2), 54.7 (OCH3-1), 53.7 (C-4),
36.9 (C-2%), 17.7 (C-6). CIMS: m/z 352 ([M−
1]−), 354 ([M+1]+). Anal. Calcd for
C18H27NO6: C, 61.17; H, 7.70; N, 3.96. Found:
C, 60.88; H, 7.69; N, 4.03.
1
+89° (c 0.9, CHCl3). H NMR (CDCl3): l
6.42 (d, 1 H, JNH,4 8.2 Hz, NH), 4.70 (d, 1 H,
J1,2 1.4 Hz, H-1), 4.54 (s, 2 H, CH2Ph), 3.88
(m, 1 H, H-4), 3.84 (dd, 1 H, J2,3 3.5 Hz, H-2),
3.75 (m, 2 H, H-3%a,b), 3.67 (dd, 1 H, J3,4 10.0
Hz, H-3), 3.50 (m, 1 H, H-5), 3.38 (s, 3 H,
OMe), 2.54 (t, 2 H, J 6.2 Hz, H-2%a,b), 1.18 (d,
3 H, H-6); 13C NMR (CDCl3): l 173.9 (CO),
100.4 (C-1), 73.4 (CH2Ph), 71.1 (C-3), 69.6
(C-2), 66.2 (C-3%), 66.2 (C-5), 55.0 (OMe), 54.2

E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
19
Methyl 4-azido-3-O-benzyl-4,6-dideoxy-h-
-mannopyranoside (42).—Azide 40 (12.2 g)
Methyl
amido)-2-O-methyl-h-
—A soln of the benzyl ether 37 (175 mg) in
EtOH was treated with H2 as described for the
preparation of 21 to give 26 in virtually theo-
retical yield, mp 145–146.5 °C (from acetone),
4,6-dideoxy-4-(3-hydroxypropan-
-mannopyranoside (26).
L
D
was benzylated as described for the
D
enan-
tiomer,12 to give, after chromatography, the
title 3-benzyl ether 42 (13.1 g, 74%), [h]D
1
−116° (c 1.2, MeOH). H NMR data agreed
enantiomer;12
1
with those reported for the
D
[h]D +47° (c 0.6, H2O). H NMR (D2O): l
CIMS: m/z 311 ([M+18]+). Anal. Calcd for
C14H19N3O4: C, 57.33; H, 6.53; N, 14.33.
Found: C, 57.29; H, 6.46; N, 14.30.
4.90 (d, 1 H, J1,2 1.8 Hz, H-1), 3.87–3.70 (m,
5 H, H-3,4,5,3%a,b), 3.58 (dd, 1 H, J2,3 3.0 Hz,
H-2), 3.48 (s, 3 H, OCH3-2), 3.40 (s, 3 H,
OCH3-1), 2.5 (t, 2 H, J 6.3 Hz, H-2%a,b), 1.18
Methyl 4-azido-3-O-benzyl-2-O-methyl-4,6-
13
dideoxy-h-
L
-mannopyranoside
(43).—The
(d, 3 H, J5,6 6.2 Hz, H-6); C NMR (CDCl3):
foregoing benzyl ether 42 (9.7 g) was methyl-
l 174.98 (CO), 97.82 (C-1), 79.03 (C-2), 68.10
(C-3), 67.42 (C-5), 58.91 (OCH3-2), 57.97 (C-
3%), 54.95 (OCH3-1), 38.87 (C-2%), 16.86 (C-6);
CIMS: m/z 264 ([M+1]+), 281 ([M+18]+).
Anal. Calcd for C11H21NO6: C, 50.18; H, 8.04;
N, 5.32. Found: C, 50.25; H, 8.02; N, 5.30.
ated as described for the corresponding
D
enantiomer18 to give, after chromatography,
the title 2-methyl ether 43 (9.6 g, 94%) as an
oil, [h]D −128° (c 1.3, CHCl3), lit.18, +117°
for the
D
enantiomer. The NMR data agreed
with those reported for the
D
enantiomer;18
Methyl 4,6-dideoxy-4-(3-deoxy-
tetronamido)-h- -mannopyranoside (45).— -
L
-glycero-
CIMS: m/z 325 ([M+18]+). Anal. Calcd for
C15H21N3O4: C, 58.62; H, 6.89; N, 13.67.
Found: C, 58.61; H, 6.88; N, 13.64.
L
L
Rhamnose was converted to the 4-azido
derivative 40 essentially as described by Mar-
tin et al.,23 which was then converted to the
corresponding amine 4116 whose NMR spec-
Methyl 4-amino-4,6-dideoxy-2-O-methyl-h-
-mannopyranoside (44).—The foregoing fully
L
protected compound 43 (7.3 g) was treated
with H2 as described above for similar conver-
sions to give amine 44 in virtually theoretical
yield, [h]D −50° (c 1.3, MeOH). The NMR
tra agreed with those reported for the
D
enantiomer.11–13
A mixture of the foregoing amine 41 (520
mg, 2.9 mmol), 3-deoxy- -glycero-tetronolac-
L
data agreed with those reported for the
D
tone (450 mg, 4.4 mmol) and pyridine (0.05
mL) was stirred at 100 °C until TLC (6:1:0.2
CH2Cl2 –MeOH–concd NH4OH) showed that
all starting material was consumed. The crude
product was chromatographed (twice, first us-
ing 6:1:0.2 CH2Cl2–MeOH–concd NH4OH,
then 9:1 EtOAc–MeOH), to give the title
compound 45 as a white foam in virtually
enantiomer;18 CIMS: m/z 192 ([M+1]+), 209
([M+18]+). Anal. Calcd for C8H17NO4: C,
50.25; H, 8.96; N, 7.32. Found: C, 49.95; H,
8.96; N, 7.26.
Methyl 4,6-dideoxy-4-(3-deoxy-
tetronamido) - 2 - O - methyl - h - - mannopyran-
oside (46).—A melt of amine 44 (778 mg, 4.07
mmol) and 3-deoxy- -glycero-tetronolactone
L
-glycero-
L
L
1
theoretical yield, [h]D −72° (c 1.3, H2O). H
(830 mg, 8.14 mmol) was stirred at 85 °C for 7
h. Chromatography (twice, first using 9:1
CH2Cl2–MeOH, then 1:1 CH2Cl2–acetone)
gave 46 (980 mg, 82%), mp 127–129 °C (from
NMR (D2O): l 4.72 (d, 1 H, J1,2 1.4 Hz, H-1),
4.27 (dd, 1 H, J2%,3%a 3.8, J2%,3%b 5.8 Hz, H-2%),
3.92–3.27 (m, 4 H, H-2,3,4,5), 3.72 (q, 2 H, J
5.6, J 7.4 Hz, H-4%), 3.38 (s, 3 H, OCH3), 2.02
(m, 1 H, H-3%a), 1.80 (m, 1 H, H-3%b), 1.17 (d,
3 H, J5,6 5.6 Hz, H-6); 13C NMR (CDCl3): l
177.50 (CO), 101.0 (C-1), 69.2 (C-2), 69.1
(C-2%), 68.2 (C-3), 67.3 (C-5), 58.0 (C-4%), 54.9
(O CH3), 52.9 (C-4), 36.0 (C-3%), 16.9 (C-6).
CIMS: m/z 280 ([M+1]+) and 297 ([M+
18]+). Anal. Calcd for C11H21NO7: C, 47.31;
H, 7.58; N, 5.02. Found: C, 47.05; H, 7.65; N,
5.00.
1
acetone), [h]D −61° (c 1, H2O). H NMR
(D2O): l 4.88 (d, 1 H, J1,2 1.7 Hz, H-1), 4.26
(dd, 1 H, J2%,3%a, J2%,3%b 8.7 Hz, H-2%), 3.91 (m, 1
H, H-3), 3.82–3.76 (m, 2 H, H-4,5), 3.72 (q, 2
H, J 5.8, J 7.3 Hz, H-4%a b), 3.55 (dd, 1 H, J2,3
3.4 Hz, H-2), 3.47 (s, 3 H, OCH3-2), 3.39 (s, 3
H, OCH3-1), 2.01 (m, 1 H, H-3%a), 1.80 (m, 1
13
H, H-3%b), 1.16 (d, 3 H, J5,6 5.6 Hz, H-6); C
NMR (D2O): l 177.4 (CO), 97.8 (C-1), 79.1
(C-2), 69.1 (C-2%), 67.9 (C-3), 67.1 (C-5), 58.9

20
E. Poirot et al. / Carbohydrate Research 330 (2001) 7–20
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(C-4%), 57.9 (OCH3-2), 54.9 (OCH3-1), 53.3
(C-4), 36.0 (C-3%), 16.9 (C-6). CIMS: m/z 294
([M+1]+), 311 ([M+18]+). Anal. Calcd for
C12H23NO7: C, 49.14; H, 7.90; N, 4.78. Found:
C, 49.21; H, 7.97; N, 4.76.
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323.
13. Gotoh, M.; Kova´cˇ, P. J. Carbohydr. Chem. 1994, 13,
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