Synthesis of spacer-containing di- and tri-saccharides that represent parts of the capsular polysaccharide of Streptococcus pneumoniae type 6B

Article abstract of DOI:10.1016/S0008-6215(97)00271-1

In the framework of studies towards oligosaccharide-conjugate-based vaccines against Streptococcus pneumoniae, the synthesis is reported of several spacer-containing oligosaccharides that represent parts of the capsular polysaccharide of S. pneumoniae ser

Full text of DOI:10.1016/S0008-6215(97)00271-1

Ž
.
Carbohydrate Research 306 1998 93–109
Synthesis of spacer-containing di- and
tri-saccharides that represent parts of the capsular
polysaccharide of Streptococcus pneumoniae type
6B
)
Mark J.L. Thijssen, Merlijn N. van Rijswijk, Johannis P. Kamerling ,
Johannes F.G. Vliegenthart
BijÕoet Center, Department of Bio-Organic Chemistry, Utrecht UniÕersity, P.O. Box 80.075, NL-3508
TB, Utrecht, Netherlands
Received 11 April 1997; accepted 25 August 1997
Abstract
In the framework of studies towards oligosaccharide-conjugate-based vaccines against
Streptococcus pneumoniae, the synthesis is reported of several spacer-containing oligosaccha-
rides that represent parts of the capsular polysaccharide of S. pneumoniae serotype 6B,
Ž
.
Ž
.
namely a-L-rhamnopyranosyl- 1™4 -5-O- 3-aminopropyl hydrogen phosphate -D-ribitol, 3-
Ž
.
aminopropyl D-ribitol- 5™hydrogen phosphate™2 -a-D-galactopyranoside, 3-aminopropyl
Ž
.
Ž
.
a-L-rhamnopyranosyl- 1 ™ 4 -D-ribityl- 5 ™ hydrogen phosphate ™ 2 -a-D-galactopyrano-
Ž
.
Ž
side, and 3-aminopropyl D-ribityl- 5™hydrogen phosphate™2 -a-D-galactopyranosyl- 1
.
™3 -a-D-glucopyranoside. Phosphorylations were carried out using the H-phosphonate
method. q 1998 Elsevier Science Ltd. All rights reserved.
Keywords: Phosphorylated oligosaccharide; Capsular polysaccharide; Streptococcus pneumoniae 6B; Vaccines
1. Introduction
Since the discovery of antibiotic-resistant pneumo-
have the highest incidence in pneumococcal infec-
tions in the United States, indeed is an effective
protecting agent against these infections in adults in
w x
coccal strains 1 , polysaccharide-containing vaccines
w x
the USA 2 . In many other countries, though, a
have been considered an adequate answer to bacterial
w
x
reformulation of the vaccine may be required 4–7 .
To members of high risk groups such as infants under
the age of two years, elderly people, and immuno-
compromised patients, the purified polysaccharides
from several serotypes in the vaccine are only low-
infections with Streptococcus pneumoniae. The cur-
rent polyvalent vaccine Pneumovaxw 23 2 , which is
w x
constituted of the purified capsular polysaccharides
w x
of 23 of the 90 serotypes of S. pneumoniae 3 that
w x
immunogenic 8 and, thus, do not offer the required
protection. These low-immunogenic polysaccharides
) Corresponding author.
include the capsules of the serotypes 6Ar6B, 14,
0008-6215r98r$19.00 q 1998 Elsevier Science Ltd. All rights reserved.

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)
94
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
19F, and 23F. The induction of a Thymus-Indepen-
to a carrier. More recently, we reported on the syn-
thesis of a spacer-containing tetrasaccharide repre-
senting a repeating unit of the capsular polysaccha-
Ž
.
dent TI -response by polysaccharides, which implies
the lack of immunological memory development to-
wards these antigens, and the generation of a certain
tolerance of the immune system towards the adminis-
Ž .
ride 1 of serotype 6B, namely, a-D-galactopyrano-
Ž
.
Ž
Ž
.
syl- 1™3 -a-D-glucopyranosyl- 1™3 -a-L-rhamno-
w x
.
Ž
tered polysaccharides 9 , also add to the desire for
pyranosyl- 1 ™ 4 -5-O- 3-aminopropyl hydrogen
.
w x
the development of more effective vaccines. Polysac-
charide–protein conjugates have proven their value in
that they are able to convert the TI-response into a
phosphate -D-ribitol 27 . Here, we describe the
preparation of a series of spacer-containing di- and
Ž
.
trisaccharides 38, 43, 47, and 51 related to the 6B
capsular polysaccharide, all having a phosphate func-
tion.
Ž
.
Thymus-Dependent TD -response, with the con-
comitant development of immunological memory
w
x
10–17 . However, antibodies produced against this
kind of conjugate are often less effective in affording
w
.
Ž
.
™2 -a-D-Galp- 1™3 -
1
Ž
.
Ž
.
a-D-Glcp- 1™3 -a-L-Rhap- 1™4
w
x
protection 18 . Vaccines consisting of oligosaccha-
ride-conjugates are now being investigated for their
ability to overcome these disadvantages 19–22 . The
required size of oligosaccharides needed for an ade-
quate immune response can be as small as one disac-
charide repeating unit. For Haemophilus influenzae
type b, di- and tetra-saccharide conjugates one or
two repeating units were able to bind polysaccha-
ride-specific antibodies if administered in sufficient
amounts 23 . A tetrasaccharide repeating unit of the
capsular polysaccharide of S. pneumoniae type 23F
conjugated to KLH was shown to elicit antibodies
with no major difference in epitope specificity com-
pared to antibodies elicited against the native poly-
Ž
x
-D-Rib-ol- 5™phosphate™
Ž
.
Ž
a-L-Rhap- 1™4 -D-Rib-ol- 5™phosphate
38
43
47
51
w
x
Ž
.
Ž
™ CH2 3NH2
.
D-Rib-ol- 5™phosphate™2 -a-D-Galp-
Ž
Ž
.
1™O CH2 3NH2
Ž
.
Ž
a-L-Rhap- 1™4 -D-Rib-ol- 5™phosphate
Ž
.
Ž
Ž
.
.
™2 -a-D-Galp- 1™O CH2 3NH2
.
Ž
D-Rib-ol- 5™phosphate™2 -a-D-Galp-
Ž
.
Ž
Ž
.
1™3 -a-D-Glcp- 1™O CH2 3NH2
w
x
2. Results and discussion
In order to produce building blocks that can be
used in the preparation of the aimed di- and tri-sac-
charide products with a 3-aminopropyl spacer, syn-
thetic routes were devised towards suitably protected
w
x
saccharide 21 .
In previous reports, we have presented the synthe-
sis of various oligosaccharide fragments of the capsu-
lar polysaccharides of the cross-reactive serotypes 6A
and 6B 24–26 , which strains are among the most
virulent serotypes of S. pneumoniae. These com-
pounds, however, could not be used in immunologi-
cal studies towards oligosaccharide-conjugate vac-
cines because of the lack of a spacer unit for coupling
Ž
.
derivatives of a-L-rhamnopyranosyl- 1™4 -D-ribitol
w
x
Ž
.
Ž
.
™5 , D-ribitol ™8 , 3-aminopropyl a-D-galacto-
Ž
Ž
.
.
pyranoside ™12 , and 3-aminopropyl a-D-galacto-
Ž
.
pyranosyl- 1 ™ 3 -a-D-glucopyranoside ™ 33 .
Combination of the appropriate fragments by phos-
phorylation and subsequent deprotection will result in
Scheme 1.

(
)
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
95
the target compounds 38, 43, 47, and 51. The protec-
tive group patterns used allow for future expansion of
the strategy towards spacer-containing tetrasaccharide
derivatives.
was estimated from the 13C NMR spectrum to be 1:1.
Ž
.
The use of iodonium dicollidineperchlorate IDCP
w
x
32 instead of methyl triflate gave a better a:b ratio
Ž
.
Ž
.
3:2 but lowered the yield of 11 51% . Separation
Preparation of building blocks.—The first build-
of the a and b anomers was possible only after
Ž
.
5
ing block 2,3,4-tri-O-acetyl-a-L-rhamnopyranosyl -
Ž .
de-p-methoxybenzylation using ammonium
Ž
.
Ž
.
1 ™ 4 -1-O-benzoyl-2,3-di-O-benzyl-D-ribitol
cerium IV nitrate, to give the desired compound 12
Ž
.
Ž
.
was prepared in two steps by coupling of ethyl
58% , and 12b 41% .
Ž .
2
2,3,4-tri-O-acetyl-1-thio-a-L-rhamnopyranoside
28 and 5-O-allyl-1-O-benzoyl-2,3-di-O-benzyl-D-
In an alternative reaction sequence, ethyl 1-thio-b-
w
x
Ž
.
w
x
D-galactopyranoside 13 33,34 was converted into
Ž .
3
w x
Ž
.
ribitol
27 in 1,2-dichloroethane-diethyl ether
14 71% using 2,2-dimethoxypropane, affording a
Ž
.
Ž
.
using N-iodosuccinimide NIS –triflic acid TfOH
galactose moiety with a HO-2 group, that could be
w
x
Ž
.
Ž
. Ž
.
29 ™4, 74% , and subsequent deallylation of the
allylated ™15, 67% Scheme 2 . Deisopropylide-
Ž
resulting disaccharide Wilkinson catalyst, then mer-
nation of 15 using aqueous 50% CH3COOH at 50 8C
. Ž
Ž
Ž
.
curic oxide–mercuric chloride;™5, 68% Scheme
™16, 98% , followed by benzylation, yielded 17
.
.
70% . Coupling of 17 with spacer 9 using iodonium
1 .
In order to obtain 1,2,3,4-tetra-O-benzyl-D-ribitol
Ž . Ž .
w
x
8 , 5-O-allyl-2,3-di-O-benzyl-D-ribitol 6 30 was
Ž
.
benzylated ™7, 90% and subsequently deallylated
by stirring with potassium t-butoxide in DMF at 80
8C followed by cleavage of the resulting 1-propenyl
Ž
.
Ž
.
function ™8, 83% .
in a higher total yield 99% . Separation of the
Preparation of the third building unit 3-N-benzyl-
anomers of 18 was only possible after removal of the
Ž
.
oxycarbonylaminopropyl 3,4,6-tri-O-benzyl-a-D-
protecting group at C-2, so that 12 38% and 12b
Ž
.
Ž
. Ž
galactopyranoside 12 started with coupling of ethyl
3,4,6-tri-O-benzyl-2-O-p-methoxybenzyl-1-thio-b-D-
32% were isolated after deallylation of 18 Wilkin-
.
son catalyst, then mercuric oxide–mercuric chloride .
In an attempt to improve the a:b ratio in the
preparation of the 3-aminopropyl a-D-galactopyrano-
side building block, by reduction of the reactivity of
the donor molecule, another reaction scheme was
Ž
. w x
galactopyranoside 10 27 with 3-N-benzyloxycar-
Ž .
w x
bonylaminopropanol 9 31 in dichloromethane–di-
ethyl ether using methyl triflate MeOTf as a pro-
moter, giving 11 62% Scheme 2 . The a:b ratio
Ž
.
Ž
. Ž
.
Scheme 2.

(
)
96
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
Ž
.
followed. Instead of benzyl, acetyl protective groups
were introduced on the proper positions in the
molecule.
Scheme 3 . First, 23 was prepared by glycosylation
of spacer 9 with ethyl 3-O-allyl-2,4,6-tri-O-benzyl-1-
Ž
. w x
thio-b-D-glucopyranoside 21 24 in diethyl ether
using methyl triflate, to give 22 52% and 22b
Ž .
Thus, 15 was deisopropylidenated using aqueous
Ž
.
Ž
.
50% CH3COOH at 50 8C ™16 , then acetylated to
30% . A coupling procedure using iodonium dicol-
lidineperchlorate in dichloromethane–diethyl ether
Ž
. Ž
.
give 19 82% over 2 steps Scheme 2 . Coupling of
Ž
.
this donor to spacer 9 in dichloromethane-diethyl
gave 22ab in a yield of 62% a:b 3:1 . Subsequent
removal of the allyl function of 22 Wilkinson cata-
.
lyst, then mercuric oxide–mercuric chloride gave
compound 23 91% .
Ž
ether using methyl triflate as a promoter gave 20
1
Ž
.
61% , with an a:b ratio of 1:2, as indicated by H
and 13C NMR analysis. Attempted coupling of 19 to
9 using iodonium dicollidineperchlorate resulted in a
complex reaction mixture.
Ž
.
In order to prepare a suitable galactosyl donor,
p-methoxyphenyl 2,3,4,6-tetra-O-acetyl-b-D-galacto-
Ž
.
w x
Ž
.
The ease of preparation of 17 and the better cou-
pling results make this donor superior to 10 and 19 in
the preparation of the protected Gal-spacer analogues,
despite the modest yield of deallylation of 18.
The preparation of building block 3-N-benzyl-
pyranoside 24 35 was deacetylated ™25 , and
Ž
.
then isopropylidenated ™26, 75% over 2 steps .
Ž
.
Allylation of 26 with allyl bromide ™27 and sub-
sequent deisopropylidenation in aqueous 50%
Ž
.
CH3COOH at 50 8C gave 28 97% from 26 , which
was acetylated to give 29 quant. . Removal of the
p-methoxyphenyl function using ammonium
cerium IV nitrate proved to be unexpectedly 35–37
difficult, resulting in a modest yield for 30 47% .
Activation of the anomeric centre with trichloroace-
tonitrile in the presence of potassium carbonate gave
Ž
Ž
.
oxycarbonylaminopropyl 3,4,6-tri-O-acetyl-a-D-
. Ž
.
galactopyranosyl - 1™3 -2,4,6-tri-O-benzyl-a-D-
Ž
.
Ž
.
w
x
glucopyranoside 33 could be accomplished via a
stepwise route, by glycosylating 3-N-benzyloxycar-
bonylaminopropyl 2,4,6-tri-O-benzyl-a-D-gluco-
Ž
.
Ž
.
pyranoside 23 with a suitable galactosyl donor
Scheme 3.

(
)
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
97
Scheme 4.
Ž
.
Ž
.
imidate 31 as a mixture of anomers a:b 1:1, 97% .
Glycosylation of glucosyl acceptor 23 with imidate
donor 31 in diethyl ether using trimethylsilyl triflate
obtained in a yield of only 26% and 32b, 27% .
Removal of the allyl function at O-2 of the galactose
residue Wilkinson catalyst, then mercuric oxide–
Ž
Ž
.
.
Ž
.
as a promoter gave disaccharide derivative 32 67% ,
and 32b 28% . When using thioethyl donor 19 with
mercuric chloride gave 33 64% .
Ž
.
Preparation and deprotection of phosphorylated
methyl triflate as a promoter in diethyl ether, 32 was
compounds.—The phosphate function was intro-
Scheme 5.

(
)
98
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
Scheme 6.
duced in all molecules via the phosphonate method
For the preparation of disaccharide 3-aminopropyl
w
x
Ž
38–43 . Thus, after preparation of the phosphonate
glycoside 43, two pathways were followed Scheme
Ž
.
Ž
ester of 5 2-chloro-4H-1.3.2-benzodioxaphosphorin-
5 . First, 12 was phosphonylated 2-chloro-4H-
.
4-one in pyridine–acetonitrile,™34, 68% , phospho-
1.3.2-benzodioxaphosphorin-4-one in pyridine–aceto-
.
nylation of spacer 9 in pyridine–acetonitrile with 34
nitrile,™39, quant and then coupled with 8 in pyri-
w
x
Ž
.
Ž
.
in the presence of pivaloyl chloride 41 ™35 , and
subsequent mild in situ oxidation of the resulting
phosphonate diester using iodine in pyridine–water,
dine–acetonitrile using pivaloyl chloride ™41 , fol-
lowed by mild in situ oxidation using iodine in
Ž
.
pyridine–water, giving 42 38% over 2 steps . Alter-
Ž
. Ž
.
Ž
gave 36 43% over 2 steps Scheme 4 . Deprotection
natively, compound 8 was phosphonylated 2-chloro-
of the resulting phosphate diester by deacylation
Ž
.
methanol–ammonia,™37 , followed by removal of
Ž
the benzyl and benzyloxycarbonyl functions Pd–C,
H2 gave, after desalting on Bio-Gel P-2, target
.
Ž
.
structure 38 in a yield of 74% from 36 .
Scheme 7.

(
)
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
99
moval of the benzyl and benzyloxycarbonyl groups
tus. Evaporations were conducted under reduced
Ž
Ž
.
Ž
.
Pd–C, H2 and desalting on Bio-Gel P-2 gave 43
pressure at 40 8C bath . Reactions were performed
under dry conditions using an atmosphere of nitrogen
or argon.
.
77% .
For the preparation of trisaccharide 3-aminopropyl
Ž
.
(
. Ž
glycoside 47 Scheme 6 , disaccharide derivative 5
was coupled with compound 39 in pyridine–aceto-
nitrile using pivaloyl chloride to give 44, which was
mildly oxidised in situ with iodine in pyridine–water
2,3,4-Tri-O-acetyl-a-L-rhamnopyranosyl - 1 ™
.
4 -5-O-allyl-1-O-benzoyl-2,3-di-O-benzyl-D-ribitol
Ž .
4 .—A mixture of ethyl 2,3,4-tri-O-acetyl-1-thio-a-
Ž .
w
x
Ž
.
3
L-rhamnopyranoside 2 28 1.03 g, 3.08 mmol ,
5-O-allyl-1-O-benzoyl-2,3-di-O-benzyl-D-ribitol
Ž
.
Ž
Ž .
to give 45 89% . Deacylation methanol–
˚
.
w
x
Ž
.
ammonia,™46 , followed by removal of the benzyl
27 1.32 g, 2.77 mmol and 4 A molecular sieves in
Ž
.
Ž
.
and benzyloxycarbonyl functions Pd–C, H2 , and
1,2-dichloroethane 8.0 mL was stirred for 30 min at
Ž
Ž
.
.
desalting on Bio-Gel P-2, gave 47 47% over 2
0 8C. Then, a mixture of NIS 0.70 g, 3.11 mmol
and TfOH 25 mL, 0.28 mmol in 1:1 1,2-dichloro-
ethane–Et2O 25 mL was added. After 1 min, TLC
9:1 toluene–EtOAc showed the appearance of a
single product Rf 0.54 , and the mixture was neu-
tralised with Et3N, filtered through Celite, diluted
with CH2Cl2, washed with aq 10% Na2S2O3 2=
and water 3= , dried MgSO4 , filtered, and con-
centrated. Column chromatography 9:1 toluene–
EtOAc of the residue afforded 4, isolated in 74% as
a syrup 1.53 g ; a D y258 c 1 ; H NMR CDCl3 :
d 8.05–7.10 m, 15 H, 3 Ph , 5.834 m, 1 H,
.
Ž
steps .
Ž
.
Finally, for the preparation of trisaccharide 3-
aminopropyl glycoside 51, compound 40 was coupled
with disaccharide derivative 33 in pyridine–aceto-
nitrile using pivaloyl chloride ™48 , followed by
mild in situ oxidation of the resulting phosphonate
diester using iodine in pyridine–water, giving 49
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
. Ž
.
Ž
77% over 2 steps Scheme 7 . Deprotection as
Ž
.
described above methanol–ammonia, ™50; then
1
.
Ž
Ž
.
Ž
w x
Ž
.
.
Ž
.
Pd–C, H2 gave, after desalting on Bio-Gel P-2,
.
Ž
product 51 quant. over 2 steps .
X
.
Ž
.
X
X
Conjugation of the free, spacer-containing saccha-
rides to carrier proteins and the results of immunolog-
ical tests with the neoglycoconjugates will be pub-
lished elsewhere.
OCH2CHsCH2 , 5.148 d, 1 H, J1 ,2 1.9 Hz, H-1 ,
X
Ž
.
X
X
X
X
5.045 t, 1 H, J3 ,4 sJ4 ,5 s9.8 Hz, H-4 , 2.143,
Ž
.
Ž
2.000, and 1.992 3 s, each 3 H, 3 Ac , 1.001 d, 3 H,
X
.
X
X
J5 ,6 6.2 Hz, 3 H-6 . Anal. Calcd for C41H48O13: C,
65.76; H, 6.46. Found: C, 66.02; H, 6.42.
Ž
. Ž
2,3,4-Tri-O-acetyl-a-L-rhamnopyranosyl - 1 ™
3. Experimental
.
Ž .
4 -1-O-benzoyl-2,3-di-O-benzyl-D-ribitol 5 .—To a
soln of
diazabicyclo 2.2.2 octane DABCO; 400 mg, 3.57
mmol in 8:3:1 EtOH–toluene–water 120 mL was
added tris triphenylphosphine rhodium I chloride
250 mg . After boiling under reflux for 45 min, TLC
1
Ž
Ž .
1.40 g, 1.86 mmol and 1,4-
General methods.— H NMR spectra 300 and 500
4
w
.
x
Ž
MHz were recorded at 25 8C with Bruker AC 300
and Bruker AMX 500 spectrometers. 13C NMR spec-
.
Ž
.
Ž
.
Ž
.
Ž .
tra 50 and 75 MHz were recorded at 25 8C with
Ž
Ž
.
Bruker WP 200 and Bruker AC 300 spectrometers.
31
Ž
.
.
P NMR spectra 121 MHz were recorded at 25 8C
9:1 toluene–EtOAc showed a complete conversion
with a Bruker AC 300 spectrometer. Chemical shifts
of the allyl into the 1-propenyl function. The mixture
was diluted with CH2Cl2, washed with 1 M HCl and
1
Ž .
d are given in ppm, for H relative to the signal for
Ž
.
Ž
.
internal Me4Si CDCl3 or internal sodium 4,4-di-
water 3= , and concentrated. To a soln of the
Ž
Ž
.
Ž
methyl-4-silapentane-1-sulfonate D2O; indirectly to
residue in 9:1 acetone–water 50 mL were added
HgO 670 mg, 3.09 mmol and HgCl2 780 mg, 2.87
mmol . After stirring the mixture for 30 min, TLC
9:1 toluene–EtOAc showed the depropenylation to
internal acetone, d 2.225 , for 13C relative to the
.
Ž
.
.
signal for internal CDCl3 d 76.9 , and for 31P
relative to the signal for external aq 85% H3PO4.
Column chromatography was performed on Kieselgel
Ž
.
Ž
.
be completed. The mixture was diluted with CH2Cl2,
Ž
.
60 E. Merck, -230 mesh or 70–230 mesh and
fractions were monitored by TLC on Kieselgel 60
Ž
.
F254 E. Merck , using detection with UV light and
then charring with aq 50% H2SO4. Optical rotations
were measured at 20 8C for solutions in CHCl3
unless stated otherwise with a Perkin–Elmer 241
polarimeter, using a 10-cm 1-mL cell. Melting points
Ž
.
Ž
.
uncorrected were determined with a Kofler appara-

(
)
100
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
.
Ž
J5a,5b 12.1 Hz, H-5a , 4.491 dd, 1 H, J4,5b 5.0 Hz,
3-N-Benzyloxycarbonylaminopropyl 3,4,6-tri-O-
benzyl-2-O-p-methoxybenzyl-arb-D-galactopyranoside
.
Ž
.
H-5b , 2.144, 2.006, and 2.000 3 s, each 3 H, 3 Ac ,
1.033 d, 3 H, J5 ,6 6.2 Hz, 3 H-6 ; 13C, d 169.9–
X
Ž
Ž
.
X
X
Ž
.
11 and 3-N-benzyloxycarbonylaaminopropyl 3,4,6-
X
.
Ž
.
Ž
.
Ž .
169.6 COCH3 and COPh , 96.2 C-1 , 78.7, 76.8,
76.6, 70.7, 69.9, 69.1, and 67.0 C-2,3,4,2 ,3 ,4 ,5 ,
73.6 and 72.4 2 OCH2 Ph , 63.4 and 61.2 C-1,5 ,
tri-O-benzyl-a-D-galactopyranoside 12 .— a
A
X
X
X
X
Ž
.
.
mixture of ethyl 3,4,6-tri-O-benzyl-2-O-p-methoxy-
Ž
Ž
.
Ž
.
Ž
. w x Ž
benzyl-1-thio-b-D-galactopyranoside 10 27 155
X
. Ž
.
Ž
.
20.6 and 20.5 2 C 3 COCH3 , 17.0 C-6 . Anal.
Calcd for C38H44O13 P1r2H2O: C, 63.59; H, 6.32.
Found: C, 63.79; H, 6.45.
mg, 0.252 mmol , 3-N-benzyloxycarbonyla-
minopropanol 9 31 69 mg, 0.33 mmol and 4 A
molecular sieves in 1:5 CH2Cl2–Et2O 9 mL was
˚
Ž .
w
x
Ž
.
Ž
.
Ž .
5-O-Allyl-1,2,3,4-tetra-O-benzyl-D-ribitol 7 .—A
soln of 5-O-allyl-2,3-di-O-benzyl-D-ribitol 6 30
2.0 g, 5.4 mmol and benzyl bromide 1.53 mL, 12.8
mmol in DMF 25 mL was added dropwise to a
stirred, cooled 0 8C suspension of NaH 0.38 g,
15.8 mmol in DMF 10 mL . TLC 8:2 hexane–
EtOAc, Rf 0.65 indicated the benzylation to be
completed in 1 h. After destroying the excess of NaH
cooled to y40 8C, and stirred for 30 min. Then,
Ž . w x
Ž
.
MeOTf 70 mL, 0.62 mmol was added. After 2 h,
Ž
.
Ž
TLC did not show any reaction, and the temperature
.
Ž
.
Ž
was raised to 0 8C. After 12 h, TLC 6:4 hexane–
Ž
.
Ž
.
EtOAc indicated the disappearance of 10 and the
appearance of a product with Rf 0.79 11 . The
.
Ž
.
Ž
Ž
.
.
mixture was neutralised with Et3N, diluted with
Ž
.
Ž
.
CH2Cl2, washed with water 3= , dried MgSO4 ,
with MeOH, the mixture was diluted with EtOAc,
filtered, and concentrated. Column chromatography
Ž
.
Ž
.
Ž
.
washed with water 3= , dried MgSO4 , filtered,
6:4 hexane–EtOAc of the residue afforded 11, iso-
Ž
Ž
. Ž .
and concentrated. Column chromatography 8:2 hex-
ane–EtOAc of the residue gave 7, isolated in 90%
yield as a syrup 2.66 g ; a q38 c 1 ; H NMR
CDCl3 : d 7.32–7.26 m, 20 H, 4 Ph , 5.881 m, 1
H, OCH 2C H5CH 2
OCH2CH5CH2 , 4.724, 4.707, 4.679, 4.656, 4.624,
and 4.594 6 d, each 1 H, 3 OCH2 Ph , 4.487 s, 2 H,
lated as a syrup 119 mg, 62%, a:b 1:1 . b A
mixture of 10 250 mg, 0.407 mmol , 9 105 mg,
0.502 mmol , and 4 A molecular sieves in 1:5 1,2-di-
chloroethane–Et2O 4.2 mL was stirred for 30 min.
Then, IDCP 0.39 g, 0.83 mmol was added. After 20
min, TLC 7:3 hexane–EtOAc indicated the disap-
pearance of 10 and the appearance of a new spot 11,
Rf 0.32 . The mixture was diluted with CH2Cl2 ,
.
Ž
.
Ž
1
˚
Ž
.
w x
Ž
.
.
D
Ž
.
Ž
.
Ž
Ž
.
.
Ž
Ž
.
.
,
5.27–5.10 m,
2
H,
.
Ž
Ž
.
Ž
Ž
.
Ž
.
.
OCH2 Ph , 3.95–3.93 m, 2 H, OCH2CH5CH2 .
Anal. Calcd for C36H40O5: C, 78.23; H, 7.29. Found:
C, 78.09; H, 7.21.
filtered through Celite, washed with aq 10%
Na2S2O3, water, aq 10% NaHCO3, and water, dried
Ž .
Ž
.
1,2,3,4-Tetra-O-benzyl-D-ribitol 8 .—A soln of 7
MgSO4 , filtered, and concentrated. Column chro-
Ž
.
Ž
Ž
.
Ž
.
2.60 g, 4.70 mmol in DMF 50 mL was heated at
80 8C, and KOtBu 620 mg, 5.52 mmol was added,
giving the soln a deep black colour. After 30 min,
TLC 25:1 toluene–acetone indicated the complete
conversion of the allyl Rf 0.69 into the 1-propenyl
function Rf 0.83 . The mixture was cooled, diluted
with CH2Cl2, washed with water 3= , and concen-
trated. The residue was dissolved in 9:1 acetone–0.1
M HCl 40 mL and the soln was boiled under reflux
for 30 min, when TLC 25:1 toluene–acetone indi-
cated a complete conversion of the 1-propenyl-con-
taining compound into a lower moving spot Rf
0.23 . The mixture was neutralised with aq 25%
NH4OH, concentrated, diluted with CH2Cl2, washed
with aq 10% NaHCO3 and water, dried MgSO4 ,
filtered, and concentrated. Column chromatography
25:1 toluene–EtOAc of the residue afforded 8, iso-
matography 9:1 toluene–acetone of the residue af-
forded 11, isolated as a syrup 159 mg, 51%, a:b
3:2 ; C NMR CDCl3 : d 156.4 NCOOCH2 Ph ,
103.3 C-1b , 98.0 C-1 a , 79.0, 75.8, 75.1, and
69.7 C-2,3,4,5 , 74.4, 73.1, and 72.9 3 OCH2 Ph ,
69.1 and 66.2 2 C NCOOCH2 Ph, OCH2 CH2 2 N,
and C-6 , 55.0 C6H4OCH3 , 39.4 O CH2 2CH2 N ,
29.4 OCH2CH2CH2 N .
.
Ž
13
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
. Ž
Ž
.
Ž
.
.
Ž
.
w
Ž
.
x
Ž
.
Ž
.
Ž .
Ž
.
a To a soln of 11 200 mg, 0.262 mmol in 3:6:1
Ž
.
Ž
.
toluene–acetonitrile–water 6 mL was added ammo-
nium cerium IV nitrate CAN; 320 mg, 0.584 mmol .
After stirring vigorously for 45 min, TLC 8:2
CH2Cl2–EtOAc indicated the conversion of 11 into
12 Rf 0.24 to be completed. The mixture was
diluted with CH2Cl2, washed with water, aq 10%
NaHCO3, and water, dried MgSO4 , filtered, and
concentrated. Column chromatography 75:25
CH2Cl2–EtOAc of the residue afforded 12b 69
mg, 41%, Rf 0.49 and 12 98 mg, 58%, Rf 0.39 ,
both isolated as a syrup. b To a soln of 18 213 mg,
0.312 mmol and DABCO 155 mg, 1.38 mmol in
Ž
.
Ž
.
Ž
Ž
.
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
1
Ž
.
w x
Ž
.
.
Ž
lated in 83% as a syrup 2.01 g ; a D q68 c 1 ; H
Ž
.
Ž
.
.
Ž
.
NMR CDCl3 ; d 7.50–7.15 m, 20 H, 4 Ph , 4.728,
Ž
Ž .
Ž
4.708, 4.662, 4.623, 4.518, and 4.472 6 d, each 1 H,
.
Ž
.
Ž
.
Ž
.
3 OCH2 Ph , 4.565 s, 2 H, OCH2 Ph , 2.27 bt, 1 H,
.
Ž
.
HO-5 .
8:3:1 EtOH–toluene–water 24 mL was added

(
)
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
101
Ž
.
Ž .
Ž
.
tris triphenylphosphine rhodium I chloride 70 mg .
of the residue gave 15, isolated in 67% as a syrup
1
Ž
Ž
.
w x
Ž
.
Ž
.
After boiling under reflux for 18 h, TLC 7:3 hex-
ane–EtOAc indicated a complete conversion of the
allyl into the 1-propenyl function Rf 0.51 . The
mixture was diluted with CH2Cl2, washed with 0.1
M HCl and water, and concentrated. The residue was
dissolved in 9:1 acetone–water 20 mL , and to the
soln were added HgCl2 500 mg, 1.84 mmol and a
catalytic amount of HgO 5.0 mg . After stirring the
mixture for 18 h, TLC 75:25 hexane–EtOAc indi-
cated the depropenylation to be completed. The mix-
ture was diluted with CH2Cl2, washed with water, aq
5% KI, water, aq 10% NaHCO3, and water, dried
0.75 g ; a y128 c 1 ; H NMR CDCl3 : d
D
.
Ž
.
Ž
5.954 m, 1 H, OCH2CH5CH2 , 5.34–5.17 m, 2 H,
OCH2CH5CH2 , 4.373 d, 1 H, J1,2 9.7 Hz, H-1 ,
4.29–4.24 m, 2 H, OCH2CH5CH2 , 4.208 dd, 1
H, J3,4 5.6, J4,5 2.0 Hz, H-4 , 4.136 dd, 1 H, J2,3
6.5 Hz, H-3 , 3.798 m, 1 H, J5,6a sJ5,6b s6.2 Hz,
H-5 , 3.398 dd, 1 H, H-2 , 3.222 s, 3 H,
C CH3 2OCH3 , 2.79–2.63 m, 2 H, SCH2CH3 ,
1.352 and 1.343 2 s, each 3 H, C CH3 2OCH3 ,
Ž
.
.
Ž
.
Ž
.
Ž
.
Ž
Ž
.
.
Ž
Ž
.
.
Ž
.
.
Ž
Ž
.
Ž
.
Ž
.
.
Ž
.
Ž
Ž
.
Ž
.
1.304 t, 3 H, SCH2CH3 . Anal. Calcd for
C18H32O6S: C, 57.42; H, 8.57. Found: C, 57.33; H,
8.66.
Ž
.
MgSO4 , filtered, and concentrated. Column chro-
Ethyl 2-O-allyl-3,4,6-tri-O-benzyl-1-thio-b-D-
Ž
.
Ž
.
Ž
matography 75:25 CH2Cl2–EtOAc of the residue
afforded 12b 64 mg, 32%, Rf 0.49 and 12 77 mg,
38%, Rf 0.39 , both isolated as a syrup; for 12: a
q198 c 1 ; NMR CDCl3 : H, d 7.60–7.10 m, 20
H, 4 Ph , 5.07 bs, 2 H, COOCH2 Ph , 4.924 d, 1 H,
J1,2 4.0 Hz, H-1 , 4.860, 4.679, 4.628, 4.533, 4.510,
and 4.402 6 d, each 1 H, 3 OCH2 Ph , 4.167 ddd, 1
galactopyranoside 17 .—A soln of 15 4.0 g, 11
Ž
.
.
Ž
.
Ž
.
mmol in aq 50% HOAc 250 mL was stirred for 18
h at 50 8C, when TLC 85:15 CH2Cl2–acetone
w x
Ž
.
D
1
Ž
.
Ž
.
Ž
indicated the deisopropylidenation to be completed.
The mixture was concentrated, and co-concentrated
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
with toluene 2= , EtOH 2= , and CH2Cl2 2= ,
Ž
.
Ž
Ž
.
giving 16 as a syrup 2.75 g, 98% . A soln of 16
2.30 g, 8.70 mmol and benzyl bromide 3.7 mL, 31
mmol in DMF 15 mL was added dropwise to a
stirred, cooled 0 8C suspension of NaH 1.55 g,
64.6 mmol in DMF 10 mL . TLC 7:3 hexane–
EtOAc, Rf 0.37 showed the benzylation to be com-
.
Ž
Ž
.
Ž
H, J2,3 10.0, JH-2,OH 7.1 Hz, H-2 , 3.904 dd, 1 H,
J3,4 2.7, J4,5 2.9 Hz, H-4 , 3.679 dd, 1 H, H-3 ,
.
Ž
.
.
.
Ž
.
Ž
Ž
.
Ž
Ž
.
Ž
3.25–3.14 m, 2 H, O CH2 2CH2 N , 2.36 bd, 1 H,
HO-2 , 1.90–1.74 m, 2 H, OCH2CH2CH2 N ; 13C, d
.
Ž
.
.
Ž
.
Ž
Ž
.
.
Ž
.
.
156.4 NCOOCH2 Ph , 98.6 C-1 , 79.6, 74.0, 70.0,
and 68.8 C-2,3,4,5 , 74.3, 73.3, and 72.4 3
OCH2 Ph , 69.0, 66.3, and 66.1 NCOOCH2 Ph,
OCH2 CH2 2 N, and C-6 , 38.6 O CH2 2CH2 N ,
29.2 OCH 2 C H 2CH 2 N . Anal. Calcd for
C38H43NO8: C, 71.12; H, 6.75. Found: C, 70.94; H,
6.83.
Ž
Ž
pleted in 4 h. After destroying the excess of NaH
with MeOH, the mixture was diluted with CH2Cl2,
.
Ž
Ž
Ž
.
.
w
.
x
Ž
.
Ž
.
washed with water 3= , dried MgSO4 , filtered,
Ž
.
Ž
and concentrated. Column chromatography 7:1 hex-
ane–EtOAc of the residue gave 17, as a syrup 3.25
g 70% ; a y238 c 1 ; H NMR CDCl3 : d
7.40–7.24 m, 15 H, 3 Ph , 5.986 m, 1 H,
.
Ž
1
.
w x
Ž
.
Ž
.
D
(
Ž
.
Ž
m ,
Ethyl 2-O-allyl-3,4-O-isopropylidene-6-O- 2-
)
.
Ž
methoxy-2-propyl -1-thio-b-D-galactopyranoside
Ž
side 13 33,34 3.79 g, 16.9 mmol in 2,2-di-
methoxypropane 155 mL was added a catalytic
OCH 2 C H 5 CH 2
,
5.31 – 5.13
2
H,
.
.
15 .—To a soln of ethyl 1-thio-b-D-galactopyrano-
OCH2CH5CH2 , 4.918, 4.743, 4.697, 4.588, 4.449,
and 4.400 6 d, each 1 H, 3 OCH2 Ph , 4.361 d, 1 H,
J1,2 9.3 Hz, H-1 , 4.38–4.26 m,
OCH2CH5CH2 , 3.920 d, 1 H, J3,4 2.8, J4,5 0 Hz,
Ž
.
w
x
Ž
.
Ž
.
Ž
Ž
.
.
Ž
2
H,
Ž
.
.
Ž
amount of p-toluenesulfonic acid 300 mg . TLC
Ž
.
.
Ž
.
Ž
9:1 CH2Cl2–acetone showed the formation of 14 to
H-4 , 3.687 t, 1 H, J2,3 9.3 Hz, H-2 , 3.496 dd, 1
.
Ž
.
Ž
be complete in 18 h. The mixture was neutralised
H, H-3 , 2.80–2.63 m, 2 H, SCH2CH3 , 1.275 t, 3
.
using Et3N and concentrated. Column chromatogra-
phy 91:8:1 CH2Cl2–acetone–Et3N of the residue
afforded 14 as a white foam 4.04 g 71% . A soln of
14 1.0 g, 3.0 mmol and allyl bromide 0.31 mL, 3.6
mmol in DMF 10 mL was added dropwise to a
stirred, cooled 0 8C suspension of NaH 0.15 g, 6.3
mmol in DMF 5 mL . TLC 95:5 CH2Cl2–acetone,
Rf 0.35 showed the allylation to be completed in 30
min. After destroying the excess of NaH with MeOH,
the mixture was diluted with EtOAc, washed with
water 3= , dried MgSO4 , filtered, and concen-
trated. Column chromatography 8:2 hexane–EtOAc
H, SCH2CH3 . Anal. Calcd for C32 H38O5S: C, 71.88;
Ž
.
H, 7.16. Found: C, 71.79; H. 7.09.
Ž
.
3-N-Benzyloxycarbonylaminopropyl2-O-allyl-
Ž
.
Ž
Ž
.
3,4,6-tri-O-benzyl-arb-D-galactopyranoside 18 .—
.
Ž
.
Ž .
Ž
.
Ž
a A mixture of 17 385 mg, 0.720 mmol , 9 210
mg, 1.00 mmol and 4 A molecular sieves in 1:5
CH2Cl2–Et2O 6 mL was stirred for 30 min, then
IDCP 0.67 g, 1.5 mmol was added. After 90 min,
˚
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
.
Ž
.
Ž
.
TLC 7:3 hexane–EtOAc showed the disappearance
Ž
.
of 17 and the appearance of a new spot Rf 0.25 .
The mixture was diluted with CH2Cl2, filtered
through Celite, washed with aq 10% Na2S2O3, wa-
Ž
.
Ž
.
Ž
.

(
)
102
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
Ž
.
.
Ž
.
ter, aq 10% NaHCO3, and water, dried MgSO4 ,
filtered, and concentrated. Column chromatography
11.2 Hz, H-6a , 4.077 dd, 1 H, J5,6b 6.5 Hz, H-6b ,
Ž
.
Ž
.
3.870 m, 1 H, H-5 , 3.534 t, 1 H, H-2 , 2.84–2.67
m, 2 H, SCH2CH3 , 2.133, 2.033, and 2.022 3 s,
each 3 H, 3 Ac , 1.325 t, 3 H, SCH2CH3 . Anal.
Calcd for C17H26O8S: C, 52.30; H, 6.71. Found: C,
52.19; H, 6.62.
Ž
.
Ž
.
Ž
7:3 hexane–EtOAc of the residue afforded 18, iso-
lated as a syrup 358 mg, 73%, a:b 7:5 . b A
mixture of 17 960 mg, 1.80 mmol , 9 620 mg, 2.96
mmol and 4 A molecular sieves in Et2O 50 mL
Ž
. Ž .
.
Ž
.
Ž
.
Ž
˚
.
Ž
.
was cooled to 0 8C, and stirred for 30 min. Then,
3-N-Benzyloxycarbonylaminopropyl 3,4,6-tri-O-
Ž
.
Ž
.
MeOTf 0.98 mL, 8.7 mmol was added. After 90
acetyl-2-O-allyl-arb-D-galactopyranoside 20 .—A
Ž
.
Ž
.
Ž
min, TLC 7:3 hexane–EtOAc showed the disap-
pearance of 17 and the appearance of a single spot
Rf 0.25 , and the mixture was neutralised with
Et3N, diluted with CH2Cl2, washed with water, aq
10% NaHCO3, and water, dried MgSO4 , filtered,
mixture of 19 103 mg, 0.264 mmol , 9 66 mg, 0.32
mmol and 4 A molecular sieves in 1:19 CH2Cl2–
Et2O 2 mL was cooled 0 8C and stirred for 30
min. Then, MeOTf 89 mL, 0.79 mmol was added.
After 1 h, TLC 6:4 hexane–EtOAc showed almost
no progression of the reaction, and the temperature
was raised to room temperature. After 5 h, TLC 6:4
hexane–EtOAc indicated the disappearance of 19
and the appearance of a single new spot 20, Rf
0.19 , and the mixture was neutralised with Et3N.
After dilution with CH2Cl2, the soln was washed
with aq 10% NaHCO3 2= and water, dried
MgSO4 , filtered, and concentrated. Column chro-
matography 6:4 hexane–EtOAc of the residue af-
forded 20, isolated in 61% as a syrup 86 mg, a:b
1:2 ; NMR CDCl3 : H, d 7.36–7.30 m, 5 H, Ph ,
˚
.
Ž
Ž
.
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
and concentrated. Column chromatography 65:35
.
Ž
hexane–EtOAc of the residue afforded 18 as a syrup
1.22 g, 99%, a:b 6:5 ; NMR CDCl3 : 1H, d
Ž
.
Ž
.
.
Ž
.
Ž
m ,
Ž
7.35–7.24 m, 20 H, 4 Ph , 5.951 m, 1 H,
OCH 2 C H 5 CH 2
OCH2CH5CH2 , 5.073 s, 2 H, COOCH2 Ph , 4.906
d, 1 H, J1,2 3.7 Hz, H-1, 18a , 4.450 d, 1 H, J1,2
7.2 Hz, H-1, 18b , 3.944 dd, 1 H, J2,3 9.6 Hz, H-2,
18a , 3.851 dd, 1 H, J2,3 8.4 Hz, H-2, 18b ,
.
Ž
.
,
5.30 – 5.10
2
H,
.
Ž
.
Ž
.
Ž
Ž
.
.
Ž
Ž
.
.
Ž
.
Ž
.
Ž
Ž
.
.
Ž
Ž
3.35–3.20 m, 2 H, O CH2 2CH2 N , 1.85–1.70 m,
H, OCH 2 C H2 CH 2 N ; 13 C,
d
156.4
1
.
.
Ž
.
Ž
.
2
Ž
.
Ž
.
Ž
.
Ž
NCOOCH2 Ph , 135.0 OCH2CH5CH2, 18b ,
5.826 m, 1 H, OCH2CH5CH2 , 5.41 bd, 1 H, J3,4
3.4 Hz, H-4, 20a , 5.34 bd, 1 H, J3,4 3.4 Hz, H-4,
Ž
.
.
Ž
134.8
Ž
O C H C H 5 C H , 18 a , 116.8
2
2
.
Ž
Ž
.
Ž
.
Ž
OCH2CH5CH2, 18a , 116.3 OCH2CH5CH2,
18b , 103.6 C-1, 18b , 97.9 C-1, 18a , 78.9,
76.1, 75.1, and 79.7 C-2,3,4,5 , 74.4, 73.2, 73.0,
72.2, 69.1, 67.2, and 66.1
OCH2CH5CH2, OCH2 CH2 2 N, NCOOCH2 Ph,
and C-6 , 39.2
OCH2CH2CH2 N .
Ethyl 3,4,6-tri-O-acetyl-2-O-allyl-1-thio-b-D-
galactopyranoside 19 .—A soln of 15 0.74 g, 2.0
mmol in aq 50% HOAc 50 mL was stirred for 5 h
at 50 8C, when TLC 85:15 CH2Cl2–acetone indi-
cated the deisopropylidenation to be completed. The
mixture was concentrated and co-concentrated with
20b , 5.09 bs, 2 H, COOCH2 Ph , 4.902 d, 1 H,
.
Ž
.
.
.
Ž
J1,2 3.5 Hz, H-1, 20a , 4.379 d, 1 H, J1,2 7.7 Hz,
H-1, 20b , 3.772 dd, 1 H, J2,3 10.5 Hz, H-2, 20a ,
Ž
.
.
Ž
.
Ž
Ž
Ž
.
.
Ž
3
OCH 2 Ph,
3.69–3.61 m, 2 H, OCH2 CH2 2 N , 3.501 dd, 1 H,
Ž
.
.
Ž
Ž
J2,3 10.1 Hz, H-2, 20b , 2.128, 2.026, and 2.014 3
s, each 3 H, 3 Ac, 20b , 2.116, 2.026, and 1.987 3
.
w
Ž . x
O CH 2 2 C H 2 N , 28.8
.
Ž
.
.
Ž
s, each 3 H, 3 Ac, 20a , 1.82–1.78 m, 2 H,
OCH2CH2CH2 N ; 13C, d 156.4 NCOOCH2 Ph,
.
Ž
Ž
.
Ž
.
Ž
.
20 a , 156.2 NCOOCH 2 Ph, 20 b , 134.5
OCH2CH5CH2 , 117.1 OCH2CH5CH2, 20a ,
116.4 OCH2CH5CH2, 20b , 103.4 C-1, 20b ,
97.7 C-1, 20a , 75.9, 72.2, 70.4, and 67.3 C-2,3,4,5,
.
Ž
.
Ž
.
Ž
.
.
Ž
.
Ž
.
Ž
Ž
.
.
Ž
Ž
Ž
.
20b , 73.1, 69.6, 68.3, and 66.3 C-2,3,4,5, 20a ,
Ž
.
Ž
.
Ž
.
Ž
Ž
.
.
Ž
.
toluene 2= , EtOH 2= , and CH2Cl2 2= ,
giving 16, isolated as a syrup. Compound 16 was
61.7 OCH2 CH2 2 N, 20a , 61.3 OCH2 CH2 2 N,
.
Ž
Ž
.
.
20 b , 38.9 O CH 2 2 C H 2 N, 20 b , 38.3
Ž
.
Ž Ž
.
.
Ž
dissolved in 2:1 pyridine–Ac2O 75 mL , and after
18 h, TLC 8:2 hexane–EtOAc indicated a complete
conversion of the starting material into 19 Rf 0.23 .
Column chromatography 75:25 hexane–EtOAc of
the residue afforded 19 as a syrup 629 mg, 82% ;
D y98 c 1 ; H NMR CDCl3 : d 5.879 m, 1 H,
OCH2CH5CH2 , 5.400 dd, 1 H, J3,4 3.4, J4,5 1.1
O CH2 2CH2 N, 20a , 29.5 OCH2CH2CH2 N,
20b , 28.9 OCH2CH2CH2 N, 20a , 20.4 COCH3 .
Ž
.
.
Ž
.
Ž
.
Ž
.
3-N-Benzyloxycarbonylaminopropyl 3-O-allyl-
Ž
.
Ž
.
Ž .
2,4,6-tri-O-benzyl-a-D-glucopyranoside 22 .— a A
mixture of ethyl 3-O-allyl-2,4,6-tri-O-benzyl-1-thio-
Ž
.
1
w x
a
Ž . Ž . Ž
Ž
.
.
w
x
Ž
˚
.
b-D-glucopyranoside 21 23 1.82 g, 3.40 mmol , 9
1.10 g, 5.26 mmol and 4 A molecular sieves in
Et2O 30 mL was stirred for 30 min. Then, MeOTf
1.9 mL, 17 mmol was added. After 2 h, TLC 7:3
hexane–EtOAc showed the disappearance of 21 and
the appearance of a single new spot 22, Rf 0.26 .
.
Ž
Ž
.
Ž
.
Ž
.
Hz, H-4 , 5.28–5.13. m, 2 H, OCH2CH5CH2 ,
Ž
.
Ž
Ž
Ž
.
Ž
4.965 dd, 1 H, J2,3 9.7 Hz, H-3 , 4.490 d, 1 H, J1,2
9.7 Hz, H-1 , 4.33–4.26 and 4.14–4.07 2 m, each 1
H, OCH2CH5CH2 , 4.154 dd, 1 H, J5,6a 6.7, J6a,6b
.
.
.
Ž
Ž
.

(
)
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
103
The mixture was neutralised with Et3N, diluted with
propenylation to be completed with the formation of
Ž
.
Ž
.
Ž
.
CH2Cl2, washed with water 3= , dried MgSO4 ,
a single spot 23, Rf 0.27 . The mixture was diluted
filtered, and concentrated. Column chromatography
with CH2Cl2, washed with water, aq 5% KI, water
Ž
.
Ž
.
65:35 hexane–EtOAc of the residue afforded 22
aq 10% NaHCO3, and water, dried MgSO4 , fil-
Ž
.
Ž .
Ž
.
Ž
1.20 g, 52% and 22b 0.71 g, 30% , both isolated
tered, and concentrated. Column chromatography 6:4
hexane–EtOAc of the residue afforded 23, isolated
as a syrup 1.03 g, 91% ; a q628 c 1 ; NMR
CDCl3 : H, d 7.35–7.18 m, 20 H, 4 Ph , 5.05 bs,
2 H, COOC H2 Ph , 4.810, 4.675, 4.611, 4.570, 4.507,
and 4.475 6 d, each 1 H, 3 OCH2 Ph , 4.701 d, 1 H,
Ž
.
as a syrup. b A mixture of 21 151 mg, 0.282
mmol , 9 47 mg, 0.23 mmol and 4 A molecular
sieves in 1:5 CH2Cl2–Et2O 2.4 mL was stirred for
30 min, then IDCP 210 mg, 0.448 mmol was added.
After 24 h, TLC 6:4 hexane–EtOAc indicated the
reaction to be incomplete, therefore, an additional
amount of IDCP 105 mg, 0.224 mmol in 1:5
CH2Cl2–Et2O 2.4 mL was added. TLC after an-
˚
.
Ž
.
Ž
.
w x
Ž
Ž
.
D
1
Ž
.
Ž
.
.
Ž
Ž
.
.
Ž
.
Ž
.
Ž
.
Ž
J1,2 3.6 Hz, H-1 , 4.035 t, 1 H, J2,3 sJ3,4 s9.4 Hz,
Ž
.
.
Ž
.
Ž
H-3 , 3.478 dd, 1 H, J4,5 9.0 Hz, H-4 , 3.375 dd, 1
Ž
.
.
Ž
Ž
.
.
H, H-2 , 3.25–3.14 m, 2 H, O CH2 2CH2 N , 1.82–
1.71 m, 2 H, OCH2CH2CH2 N ; 13C, d 156.4
Ž
.
other 18 h showed the disappearance of 21 and the
Ž
.
Ž
.
Ž
.
appearance of a major new spot with Rf 0.54 22 .
The mixture was diluted with CH2Cl2, filtered
through Celite, washed with aq 10% Na2S2O3 and
water 2= , dried MgSO4 , filtered, and concen-
trated. Column chromatography 65:35 hexane–
EtOAc of the residue afforded 22ab, isolated as a
syrup 95 mg, 62%, a:b 3:1 ; for 22: a D q508 c
1 ; NMR CDCl3 : H, d 7.35–7.15 m, 20 H, 4 Ph ,
5.906 m, 1 H, OCH2CH5CH2 , 5.27–5.05 m, 2 H,
OCH2CH5CH2 , 5.03 bs, 2 H, COOCH2 Ph , 4.801,
4.727, 4.577, 4.556, 4.462, and 4.445 6 d, each 1 H,
3 OCH2 Ph , 4.651 d, 1 H, J1,2 3.7 Hz, H-1 ,
4.37–4.15 m, 2 H, OCH2CH5CH2 , 3.780 t, 1 H,
J2,3 sJ3,4 s9.4 Hz, H-3 , 3.491 dd, 1 H, J4,5 9.0
Hz, H-4 , 3.454 dd, 1 H, H-2 , 3.24–3.13 m, 2 H,
NCOOCH2 Ph , 96.8 C-1 , 79.5, 77.1, 73.7, and
Ž
.
Ž
.
70.7 C-2,3,4,5 , 74.5, 73.4, and 73.0 3 OCH2 Ph ,
Ž
Ž
.
68.6, 66.2, and 66.3 C-6, OCH2 CH2 2 N, and
NCOOCH 2 Ph , 39.2 O CH 2 2C H 2 N , 29.1
OCH2CH2CH2 N . Anal. Calcd for C38H43NO8 P
1r2H2O: C, 70.13; H, 6.81. Found: C, 69.98; H,
Ž
.
Ž
.
.
w
Ž
.
x
Ž
Ž
.
.
Ž
.
w x
Ž
6.73.
1
.
Ž
.
Ž
.
p-Methoxyphenyl 2-O-allyl-b-D-galactopyranoside
28 .—To a solution of p-methoxyphenyl 2,3,4,6-te-
Ž
.
Ž
Ž
.
.
Ž
.
Ž
. w x Ž
tra-O-acetyl-b-D-galactopyranoside 24 34 7.0 g,
Ž
.
Ž
.
15 mmol in MeOH 150 mL was added NaOMe
.
Ž
.
Ž
.
Ž
pH 12 . After 18 h, TLC 10:2:1 EtOAc–EtOH–
water showed a complete conversion into 25 Rf
0.47 . The mixture was neutralised with Dowex-50
resin, filtered, and concentrated. To a solution
of the residue in 2,2-dimethoxypropane 180 mL
was added a catalytic amount of p-toluenesulfonic
acid 270 mg . After 5 h, TLC 9:1 CH2Cl2–acetone
Ž
.
Ž
.
Ž
.
Ž
.
.
q
.
Ž
.
Ž
Ž
H
Ž
.
.
Ž
Ž
.
O C H 2 C H 2 N , 1.87 – 1.68
m ,
2
H ,
2
OCH2CH2CH2 N ; 13C, d 156.4 NCOOCH2 Ph ,
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
.
135.1 OCH2CH5CH2 , 116.1 OCH2CH5CH2 ,
97.3 C-1 , 81.7, 79.4, 77.4, and 70.3 C-2,3,4,5 ,
74.8, 74.0, 73.2, and 73.0 3 OCH2 Ph and
OCH2CH5CH2 , 68.4, 66.3, and 66.2 C-6,
Ž
.
Ž
.
Ž
showed the appearance of a new spot 26, Rf 0.31 .
Ž
The mixture was neutralised with Et3N and concen-
trated. Column chromatography 90:9:1 CH2Cl2–
acetone–Et3N of the residue afforded 26, isolated as
.
Ž
Ž
Ž
.
.
.
OCH 2 CH 2 2 N, and NCOOCH 2 Ph , 39.4
w
Ž
.
x
Ž
.
Ž
.
Ž
O CH2 2CH2 N , 28.9 OCH2CH2CH2 N . Anal.
Calcd for C41H47NO8 P1r2H2O: C, 71.28; H, 7.00.
a syrup 4.59 g, 75% . A soln of 26 3.27 g, 8.21
.
Ž
.
mmol and allyl bromide 1.10 mL, 12.7 mmol in
DMF 32 mL was added dropwise to a stirred,
cooled 0 8C suspension of NaH 0.60 g, 25 mmol
in DMF 15 mL . TLC 95:5 CH2Cl2–acetone, Rf
0.43 indicated the allylation to be completed in 30
min. After destroying the excess of NaH with MeOH,
the mixture was diluted with EtOAc, washed with
water 3= , dried MgSO4 , filtered, and concen-
trated. Column chromatography 95:5 CH2Cl2–
acetone of the residue gave 27 as a syrup 3.6 g .
Compound 27 was dissolved in aq 50% HOAc 150
mL and the soln was stirred for 3 h at 50 8C, when
Ž
Ž
.
Found: C, 71.82; H, 6.79.
.
Ž
.
3-N-Benzyloxycarbonylaminopropyl 2,4,6-tri-O-
Ž
.
Ž
.
Ž
benzyl-a-D-glucopyranoside 23 .—To a soln of 22
Ž
mmol in 8:3:1 EtOH–toluene–water 120 mL was
added tris triphenylphosphine rhodium I chloride
.
Ž
.
1.20 g, 1.76 mmol and DABCO 370 mg, 3.30
.
Ž
.
Ž
.
Ž .
Ž
.
Ž
.
Ž
.
230 mg . After boiling under reflux for 8 h, the
Ž
mixture had turned black, and was diluted with
Ž
.
.
Ž
.
CH2Cl2, washed with 0.1 M HCl and water 2= ,
and concentrated. The residue was dissolved in 9:1
acetone–water 120 mL , and to the soln were added
HgCl2 2.8 g, 10 mmol and a catalytic amount of
HgO 19 mg . After stirring the mixture for 18 h,
TLC 65:35 hexane–EtOAc indicated the de-
Ž
Ž
.
.
Ž
.
Ž
.
TLC 85:15 CH2Cl2–acetone indicated the deiso-
Ž
.
Ž
.
propylidenation ™28, Rf 0.03 to be completed.
Ž
.
The mixture was concentrated and co-concentrated

(
)
104
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
Ž
.
Ž
.
Ž
.
.
Ž
.
with toluene 3= , EtOH 3= and CH2Cl2 3= .
H-3, 30a , 4.952 dd, 1 H, J2,3 10.2 Hz, H-3, 30b ,
Ž
.
Ž
.
Ž
Ž
Column chromatography 9:1 CH2Cl2–MeOH of
the residue afforded 28, isolated as a white solid
4.762 d, 1 H, J1,2 7.6 Hz, H-1, 30b , 4.448 m, 1
.
H, J5,6a sJ5,6b s6.7 Hz, H-5, 30a , 4.38–4.15 m, 2
1
Ž
.
w x
Ž
.
Ž
Ž
.
.
Ž
2.60 g, 97% ; a y148 c 1 ; H NMR CDCl3 :
H, OCH2CH5CH2 , 3.918 m, 1 H, J5,6a sJ5,6b
s
D
Ž
.
.
Ž
.
.
d 7.01–6.81 m, 4 H, C6 H4OCH3 , 5.959 m, 1 H,
6.4 Hz, H-5, 30b , 3.786 dd, 1 H, H-2, 30a , 3.513
.
Ž
Ž
.
Ž
OCH 2 C H 5 CH 2
OCH2CH5CH2 , 4.833 d, 1 H, J1,2 7.6 Hz, H-1 ,
4.515 and 4.284 2 m, each 1 H, OCH2CH5CH2 ,
3.772 s, 3 H, C6H4OCH3 , 3.611 ddd, 1 H, H-5 ,
2.95, 2.80, and 2.30 3 bs, each 1 H, 3 OH . Anal.
Calcd for C16H22O7 PH2O: C, 55.81; H, 7.02. Found:
C, 56.04; H, 6.89.
p-Methoxyphenyl 3,4,6-tri-O-acetyl-2-O-allyl-b-
D-galactopyranoside 29 .—A soln of 28 2.60 g,
,
5.35 – 5.19
m ,
2
H,
dd, 1 H, H-2, 30b , 2.147 s, 3 H, Ac, 30b , 2.141
s, 3 H, Ac, 30a , 2.055 and 2.032 2 s, each 3 H, 2
.
Ž
.
Ž
.
Ž
Ž
.
.
Ac .
Ž
.
Ž
.
Ž
.
To a solution of 30 67 mg, 0.19 mmol and
trichloroacetonitrile 163 mL, 1.63 mmol in CH2Cl2
1.5 mL was added freshly fused K2CO3 133 mg .
After 90 min, TLC 95:5 CH2Cl2–acetone showed a
complete conversion of 30 into 31 Rf 0.51 , and the
mixture was concentrated and purified over a short
silica column 9:1 CH2Cl2–acetone to give 31a 48
mg and 31b 44 mg , both isolated as a glass, in a
total yield of 97%; for 31a: a q338 c 1 ; H
NMR CDCl3 : d 6.591 d, 1 H, J1,2 3.6 Hz, H-1 ,
5.844 m, 1 H, OCH2CH5CH2 , 5.535 dd, 1 H,
J3,4 3.3, J4,5 1.4 Hz, H-4 , 5.327 dd, 1 H, J2,3 10.5
Hz, H-3 , 5.32–5.16 m, 2 H, OCH2CH5CH2 ,
4.414 m, 1 H, J5,6a sJ5,6b s6.6 Hz, H-5 , 3.974
dd, 1 H, H-2 , 2.154, 2.031, and 2.013 3 s, each 3
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
7.97 mmol in 2:1 pyridine–Ac2O 150 mL was
stirred for 18 h, when TLC 95:5 CH2Cl2–acetone
Ž
.
.
Ž
.
1
Ž
w x
Ž
.
indicated the acetylation to be completed ™29, Rf
D
.
Ž
.
Ž
.
0.61 . The mixture was concentrated and co-con-
Ž
.
Ž
.
Ž
.
Ž
Ž
centrated with toluene 3= , EtOH 3= , and
CH2Cl2 3 = . Column chromatography 95:5
CH2Cl2–acetone of the residue afforded 29, as a
syrup 3.60 g, quant ; a y38 c 1 ; H NMR
CDCl3 : d 7.02–7.00 and 6.85–6.81 2 m, each 2
H, C6 H4OCH3 , 5.890 m, 1 H, OCH2CH5CH2 ,
5.406 dd, 1 H, J3,4 3.5, J4,5 1.0 Hz, H-4 , 5.30–5.14
m, 2 H, OCH2CH5CH2 , 5.006 dd, 1 H, J2,3 10.2
Hz, H-3 , 4.866 d, 1 H, J1,2 7.7 Hz, H-1 , 4.45–4.19
m, 2 H, OCH2CH5CH2 , 4.208 dd, 1 H, J5,6b 6.8,
J6a,6b 11.5 Hz, H-6b , 4.124 dd, 1 H, J5,6a 6.4 Hz,
H-6a , 3.947 m, 1 H, H-5 , 3.784 dd, 1 H, H-2 ,
3.781 s, 3 H, C6H4OCH3 , 2.163 and 2.045 2 s, 3,6
H, 3 Ac . Anal. Calcd for C22 H28O10: C, 58.40; H,
6.24. Found: C, 58.42; H, 6.18.
Ž
.
Ž
.
.
.
Ž
.
1
Ž
.
w x
Ž
.
Ž
.
D
Ž
.
Ž
Ž
.
Ž
H, 3 Ac ; for 31b:
q48 c 1 ; 1H NMR
.
Ž
.
.
w x
a
Ž
.
D
Ž
.
Ž
Ž
.
Ž
.
CDCl3 : d 5.838 m, 1 H, OCH2CH5CH2 , 5.776
d, 1 H, J1,2 8.1 Hz, H-1 , 5.424 dd, 1 H, J3,4 3.5,
J4,5 1.3 Hz, H-4 , 5.26–5.12 m,
OCH2CH5CH2 , 5.046 dd, 1 H, J2,3 10.1 Hz,
H-3 , 4.36–4.11 m, 2 H, OCH2CH5CH2 , 4.043
ddd, 1 H, J5,6a 6.0, J5,6b 7.4 Hz, H-5 , 3.799 dd, 1
H, H-2 , 2.164, 2.044, and 2.033 3 s, each 3 H, 3
Ž
.
Ž
.
Ž
.
Ž
.
.
Ž
2
H,
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
.
Ž
.
Ž
.
Ž
.
Ž
Ž
.
Ž
.
Ž
.
.
Ac .
(
3-N-Benzyloxycarbonylaminopropyl 3,4,6-tri-O-
) (
)
3 ,4 ,6 -T ri-O -a c e ty l-2 -O -a lly l-a r b -D -
acetyl-2-O-allyl-a-D-galactopyranosyl - 1™3 -2,4,6-
Ž
.
Ž
.
Ž .
galactopyranose 30 and 3,4,6-tri-O-acetyl-2-O-al-
lyl-a r b-D-galactopyranosyl trichloroacetimidate
tri-O-benzyl-a-D-glucopyranoside 32 .— a A mix-
Ž
.
Ž
ture of 19 217 mg, 0.556 mmol , 23 241 mg, 0.376
˚
Ž
.
Ž
.
.
Ž
.
31 .—To a soln of 29 117 mg, 0.259 mmol in
mmol and 4 A molecular sieves in Et2O 10 mL
Ž
.
Ž
3:6:1 toluene–acetonitrile–water 4.0 mL was added
CAN 425 mg, 0.775 mmol . After 4 h, TLC 85:15
CH2Cl2–acetone showed the conversion of the start-
ing compound Rf 0.82 into a minor spot Rf 0.77
and a major spot 30, Rf 0.46 . The mixture was
diluted with CH2Cl2 and washed with water, aq 5%
NaHSO3 aq 10% NaHCO3, and water, dried
was stirred for 30 min. Then, MeOTf 125 mL, 1.10
Ž
.
Ž
.
Ž
mmol was added. After 7 h, TLC 9:1 CH2Cl2–
.
.
acetone showed the disappearance of 23 and the
appearance of a new spot 32, Rf 0.78 , and the
mixture was neutralised with Et3N, diluted with
CH2Cl2, washed with water 3= , dried MgSO4 ,
filtered, and concentrated. Column chromatography
95:5 CH2Cl2–acetone of the residue afforded 32
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
Ž
.
MgSO4 , filtered, and concentrated. Column chro-
Ž
.
.
Ž
.
matography 9:1 CH2Cl2–acetone of the residue
afforded 30 as a syrup 42 mg, 47%, a:b 2:1 ; H
NMR CDCl3 : d 5.882 m, 1 H, OCH2CH5CH2 ,
5.436 dd, 1 H, J3,4 3.4, J4,5 1.3 Hz, H-4, 30a ,
5.417 d, 1 H, J1,2 3.6 Hz, H-1, 30a , 5.371 dd, 1
H, J3,4 3.5, J4,5 1.2 Hz, H-4, 30b , 5.32–5.15 m, 2
H, OCH2CH5CH2 , 5.273 dd, 1 H, J2,3 10.3 Hz,
96 mg, 26% and 32b 97 mg, 27% , both isolated
1
Ž
.
Ž .
Ž
as a syrup. b A mixture of 31 145 mg, 0.295
mmol , 23 140 mg, 0.218 mmol and 4 A molecular
˚
Ž
.
Ž
.
.
.
Ž
.
Ž
Ž
.
sieves in Et2O 12.5 mL was cooled to y10 8C, and
Ž
.
Ž
Ž
Ž
stirred for 30 min. Then, TMSOTf 37 mL, 0.19
.
.
Ž
mmol was added. After 20 min, TLC 9:1 CH2Cl2–
.
Ž
.
acetone showed the disappearance of 23 and the

(
)
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
105
X
Ž
.
.
Ž
X
X
X
X
X
X
appearance of a single product Rf 0.78 , and the
mixture was neutralised with Et3N, diluted with
CH2Cl2, washed with water, aq 10% NaHCO3, and
H, J1 ,2 3.5 Hz, H-1 , 5.300 dd, 1 H, J3 ,4 3.3, J4 ,5
X
X
.
Ž
.
Ž
1.2 Hz, H-4 , 5.147 dd, 1 H, H-3 , 5.08 and 5.02 2
bd, each 1 H, COOC H2 Ph , 4.743, 4.673, 4.609,
4.534, 4.487, and 4.460 6 d, each 1 H, 3 OCH2 Ph ,
4.683 d, 1 H, J1,2 3.8 Hz, H-1 , 4.122 dd, 1 H, J2,3
9.7, J3,4 9.4 Hz, H-3 , 3.643 dd, 1 H, J4,5 9.0 Hz,
H-4 , 3.472 dd, 1 H, J2 ,3 10.5 Hz, H-2 , 3.26–3.17
m, 2 H, O CH2 2CH2 N , 2.098, 2.038, and 1.930 3
s, each 3 H, 3 Ac , 1.84–1.72 m, 2 H,
.
Ž
.
Ž
.
water, dried MgSO4 , filtered, and concentrated.
Column chromatography 93:5:6.5 CH2Cl2–acetone
of the residue afforded 32 141 mg, 67% and 32b
59 mg, 28% , both isolated as a syrup; for 32: a
q328 c 1 ; NMR CDCl3 : H, d 7.36–7.11 m, 20
H, 4 Ph , 5.659 m, 1 H, OCH2CH5CH2 , 5.545 d,
1 H, J1 ,2 3.6 Hz, H-1 , 5.368 dd, 1 H, J2 ,3 10.6,
J3 ,4 3.1 Hz, H-3 , 5.292 dd, 1 H, J4 ,5 1.5 Hz,
Ž
.
Ž
.
Ž
Ž
.
.
Ž
X
Ž
.
w x
.
Ž
.
X
X
D
1
Ž
.
Ž
.
Ž
Ž
Ž
.
.
.
Ž
.
Ž
.
Ž
Ž
X
.
Ž
X
X
X
X
.
OCH2CH2CH2 N . Anal. Calcd for C50H59NO16: C,
64.57; H, 6.39. Found: C, 64.55; H, 6.37.
X
.
Ž
Ž
X
X
X
X
X
.
.
(
) (
)
H-4 , 5.12–5.01 m, 2 H, OCH2CH5C H2 , 5.07
bs, 2 H, COOCH2 Ph , 4.730 d, 1 H, J1,2 3.7 Hz,
H-1 , 4.918, 4.629, 4.562, 4.538, and 4.448 5 d,
2,3,4-Tri-O-acetyl-a-L-rhamnopyranosyl - 1™4 -
Ž
.
Ž
(
1-O-benzoyl-2,3-di-O-benzyl-5-O- triethylammonium H
.
Ž
)
Ž
. Ž
-phosphonate -D-ribitol 34 , 2,3,4-tri-O-acetyl-a-L-
.
Ž
) (
)
1,1,1,1,2 H, 3 OCH2 Ph , 4.182 dd, 1 H, J2,3 9.7,
rhamnopyranosyl - 1™4 -1-O-benzoyl-2,3-di-O-
.
Ž
.
.
(
J3,4 8.0 Hz, H-3 , 3.707 t, 1 H, J4,5 8.0 Hz, H-4 ,
3.543 dd, 1 H, H-2 , 3.29–3.17 m, 2 H,
O CH2 2CH2 N , 2.085, 2.012, and 1.918 3 s, each 3
H, 3 Ac , 1.82–1.73 m, 2 H, OCH2CH2CH2 N ;
benzyl - 5 - O - 3 - N - benzyloxycarbonylaminopropyl
Ž
.
Ž
Ž
)
Ž
.
triethylammonium phosphate - D - ribitol 36 , a - L -
Ž
.
(
)
Ž
rhamnopyranosyl- 1™4 -2,3-di-O-benzyl-5-O- 3-N-
.
Ž
.
benzyloxycarbonylaminopropyl triethylammonium
13
Ž
.
)
Ž
.
C, d 170.2, 170.0, and 169.7 3 COCH3 , 156.4
phosphate -D-ribitol 37 , and a-L-rhamnopyranosyl-
Ž
Ž
.
Ž
.
(
)
Ž
(
)
NCOOCH2 Ph , 134.3 OCH2CH5CH2 , 117.4
1™4 -5-O- 3-aminopropyl hydrogen phosphate -D-
X
.
Ž
.
.
Ž
.
OCH2CH5CH2 , 97.7 and 96.6 C-1,1 , 78.8, 78.3,
ribitol 38 .—To a soln of 5 500 mg, 0.705 mmol
Ž
Ž
.
75.7, 73.3, 70.3, 69.9, 68.9, and 66.0 C-
in 1:5 pyridine–acetonitrile 6 mL was added 2-
chloro-4H-1.3.2-benzodioxaphosphorin-4-one 171
mg, 0.844 mmol , and the mixture was stirred for 24
h, when TLC 9:1 CH2Cl2–acetone revealed the
disappearance of the starting material. After the addi-
tion of 1:1 pyridine–water 1 mL , the mixture was
diluted with CH2Cl2, washed with 1 M triethylam-
monium bicarbonate 2= and water, dried MgSO4 ,
filtered, and concentrated. Column chromatography
80:19:1 CH2Cl2–acetone–Et3N, then 80:19:1
CH2Cl2–MeOH–Et3N of the residue afforded 34,
isolated as a syrup 421 mg, 68% ; H NMR CDCl3 :
d 7.97–6.77 m, 15 H, 3 Ph , 6.832 d, 1 H, JP – H
613 Hz, P–H , 5.355 dd, 1 H, J1 ,2 1.7, J2 ,3 3.4 Hz,
H-2 , 5.261 dd, 1 H, J3 ,4 10.2 Hz, H-3 , 5.193 d,
1 H, H-1 , 5.021 t, 1 H, J4 ,5 10.2 Hz, H-4 , 3.074
X
X
X
X
.
Ž
2,3,4,5,2 ,3 ,4 ,5 , 73.6, 73.4, 73.1, 72.9, 68.3, 66.4
X
Ž
.
Ž
.
Ž
2 C , and 61.5 3 OCH2 Ph, C-6,6 , OCH2CH5CH2,
Ž
.
.
.
.
N C O O C H P h , O C H C H 2 N , 3 8 .8
O CH2 2CH2 N , 29.3 OCH2CH2CH2 N , 20.7 and
20.5 2 C 3 COCH3 . Anal. Calcd for C53H63NO16:
C, 65.62; H, 6.55. Found: C, 65.38; H, 6.52.
2
2
2
w
Ž
.
x
Ž
Ž
.Ž
.
Ž
.
(
Ž
.
Ž
.
3-N-Benzyloxycarbonylaminopropyl 3,4,6-tri-O-
) (
)
acetyl-a-D-galactopyranosyl - 1™3 -2,4,6-tri-O-ben-
Ž
.
Ž
.
Ž
zyl-a-D-glucopyranoside 33 .—To a soln of 32 96
.
Ž
.
mg, 99 mmol and DABCO 100 mg, 891 mmol in
8:3:1 EtOH–toluene–water 24 mL was added
1
Ž
.
Ž
.
Ž
.
Ž
.
Ž .
Ž
.
.
Ž
.
Ž
tris triphenylphosphine rhodium I chloride 45 mg .
Ž
.
Ž
X
X
X
X
After refluxing for 6 h, TLC 95:5 CH2Cl2–acetone
did not show any new spots. The mixture was con-
centrated, diluted with CH2Cl2, washed with 0.1 M
HCl and water 2= , and concentrated. To a soln of
X
X
.
Ž
.
Ž
X
X
X
X
.
Ž
.
X
X
Ž
.
Ž
Ž
. .
Ž
q, 6 H, N CH2CH3 3 , 2.176, 2.126, and 2.126 3 s,
Ž
.
.
.
Ž
Ž
. .
the residue in 9:1 acetone–water 10 mL were added
each 3 H, 3 Ac , 1.333 t, 9 H, N CH2CH3 3 , 0.955
X
Ž
.
Ž
HgCl2 160 mg, 0.589 mmol and a catalytic amount
of HgO 8 mg . After stirring for 18 h, TLC 6:4
hexane–EtOAc showed the disappearance of the
d, 3 H, 3 H-6 .
Ž
.
.
Ž
Ž
.
Ž
A mixture of 34 421 mg, 0.482 mmol and 9 282
.
Ž
.
mg, 1.35 mmol in 1:5 pyridine–acetonitrile 7.9 mL
higher moving spot, indicating the rearrangement re-
action and subsequent depropenylation to be com-
pleted. The mixture was diluted with CH2Cl2, fil-
tered, washed with water, aq 5% KI, water, aq 10%
was stirred for 10 min, and pivaloyl chloride was
Ž
.
added 83 mL, 0.67 mmol . After stirring for 20 min,
Ž
an additional amount of pivaloyl chloride 50 mL,
0.41 mmol was added. After 60 min, TLC 9:1
CH2Cl2–acetone revealed the disappearance of 34
and the appearance of a new spot with Rf 0.31 35 .
.
Ž
Ž
.
.
NaHCO3, and water, dried MgSO4 , filtered, and
Ž
Ž
.
concentrated. Column chromatography 93:7
CH2Cl2–acetone of the residue afforded 33, isolated
.
Then, a 0.5 M soln of iodine in 95:5 pyridine–water
1
Ž
.
w x
Ž
.
Ž
.
as a syrup 59 mg, 64% ; a D q628 c 1 ; H NMR
100 mL was added to the mixture, and after 20 min,
Ž
.
Ž
.
Ž
Ž
.
CDCl3 : d 7.34–7.11 m, 20 H, 4 Ph , 5.441 d, 1
TLC 9:1 CH2Cl2–acetone indicated the disappear-

(
)
106
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
Ž
.
ance of 35 and a new spot on the baseline. The
mixture was diluted with CH2Cl2, washed with aq
5% Na2S2O3 and 1 M triethylammonium bicarbonate
for 4 h, when TLC 9:1 CH2Cl2–acetone indicated
an incomplete reaction. Therefore, an additional
amount of 2-chloro-4H-1.3.2-benzodioxaphosphorin-
Ž
.
Ž
.
Ž
.
2= , dried MgSO4 , filtered, and concentrated.
4-one 32 mg, 158 mmol was added, and after 20 h
Ž
Ž
.
Column chromatography 90:9:1 CH2Cl2–acetone–
Et3N, then 90:9:1 CH2Cl2–MeOH–Et3N of the
TLC 9:1 CH2Cl2–acetone revealed the disappear-
ance of the starting material. After the addition of 1:1
pyridine–water 1 mL , the mixture was diluted with
CH2Cl2, washed with 1 M triethylammonium bicar-
bonate 2= and water, dried MgSO4 , filtered, and
concentrated. Column chromatography 80:19:1
CH2Cl2–acetone–Et3N, then 80:19:1 CH2Cl2–
MeOH–Et3N of the residue afforded 39, isolated as
a glass 97 mg, quant ; H NMR CDCl3 : d 7.47–
6.74 m, 20 H, 4 Ph , 6.953 d, 1 H, JP – H 629 Hz,
P–H , 5.66–5.55 m, 2 H, COOCH2 Ph , 5.615 d, 1
H, J1,2 3.4 Hz, H-1 , 5.435, 5.334, 5.165, 5.073,
5.040, and 4.931 6 d, each 2 H, 3 OCH2 Ph , 3.029
q, 6 H, N CH2CH3 3 , 1.261 t, 9 H, N CH2CH3 3
1,2,3,4-Tetra-O-benzyl-5-O- triethylammonium
H-phosphonate -D-ribitol 40 .—To a soln of 8 83
mg, 0.16 mmol in 1:5 pyridine–acetonitrile 1.1 mL
.
Ž
.
.
Ž
.
residue gave 36, isolated as a glass 224 mg, 43% ;
13
Ž
.
Ž
Ž
C NMR CDCl3 : d 169.8 2 C and 169.6 3
X
.
Ž
.
Ž
Ž
.
Ž
.
Ž
.
COCH3 , 166.0 COPh , 96.2 C-1 , 77.2, 75.5,
75.3, 70.5, 69.5, 69.2, and 66.7 C-2,3,4,2 ,3 ,4 ,5 ,
73.0 and 72.3 2 OCH2 Ph , 66.0 and 63.7
Ž
Ž
Hz , 45.7 N CH2CH3 3 , 37.1 O CH2 2CH2 N ,
30.2 OCH2CH2CH2 N , 20.6 2 C and 20.5 3
COCH3 , 16.9 C-6 , 8.4 N CH2CH3 3 .
X
X
X
X
.
Ž
Ž
.
.
.
NCOOCH 2 Ph and C-1 , 65.0 and 62.7
OCH2 CH2 2 N and C-5, JP,OC-spacer f JP,C-5 f 5.3
1
Ž
.
Ž
.
Ž
.
.
w
Ž
.
.
x
w
Ž
.
x
Ž
Ž
.
.
Ž
Ž
Ž
.
Ž
.
Ž
X
.
Ž
Ž
.
w
Ž
. x
.
.
Ž
.
A soln of 36 21.2 mg, 19.6 mmol in 2:1 MeOH–
Ž
.
Ž
Ž
. .
Ž
Ž
. .
aq 25% NH4OH 12.5 mL was heated for 24 h at 50
8C, and concentrated, and after repeating the de-O-
acylation, crude 37 was obtained, which was purified
on Sephadex LH-20 50:50:1 CH2Cl2–MeOH–
Et3N ; C NMR CD3OD : d 101.7 C-1 , 81.2,
80.5, 77.7, 74.1, 72.4, 72.3, and 70.3 C-
2,3,4,2 ,3 ,4 ,5 , 74.7 and 73.4 2 OCH2 Ph , 67.4,
66.7, 64.3, and 62.1 NCOOCH2 Ph, OCH2 CH2 2 N,
and C-1,5 , 38.7 O CH 2 2 C H 2 N , 31.9
.
Ž
.
.
Ž
.
Ž
Ž
Ž
.
13
X
.
Ž
.
Ž
.
was added 2-chloro-4H-1.3.2-benzodioxaphosphorin-
Ž
Ž
.
4-one 70 mg, 0.35 mmol , and the mixture was
X
X
X
X
.
Ž
.
Ž
.
stirred for 2 h, when TLC 1:1 hexane–EtOAc indi-
cated the disappearance of the starting material and
the formation of a new spot on the baseline. After the
addition of 1:1 pyridine–water 0.5 mL , the mixture
was diluted with CH2Cl2, washed with 1 M trieth-
Ž
Ž
.
.
w
Ž
.
x
X
Ž
.
Ž
.
Ž
.
OCH2CH2CH2 N , 18.0 C-6 .
To a soln of 37 in 1:2:2 EtOAc–2-propanol–MeOH
Ž
.
Ž
.
Ž
.
5 mL was added 10% Pd–C 10 mg , and the
ylammonium bicarbonate 2= and water, dried
Ž
.
mixture was hydrogenolysed at atmospheric pressure
for 16 h. After filtration, the mixture was concen-
trated, and the residue was purified by Bio-Gel P-2
gel-permeation chromatography using water as elu-
ent, affording 38 as a white powder 6.3 mg, 74% ;
NMR D2O : H, d 5.074 d, 1 H, J1 ,2 1.8 Hz,
H-1 , 3.465 t, 1 H, J3 ,4 sJ4 ,5 s9.6 Hz, H-4 ,
3.156 t, 2 H, O CH2 2CH2 N , 2.11–1.98 m, 2 H,
OCH2CH2CH2 N , 1.292 d, 3 H, J5 ,6 6.3 Hz, 3
H-6 ; C, d 101.6 C-1 , 78.4 C-4, JP,C-4 7.6 Hz ,
MgSO4 , filtered, and concentrated. Column chro-
matography 80:19:1 CH2Cl2–acetone–Et3N, then
80:19:1 CH2Cl2–MeOH–Et3N of the residue af-
forded 40, isolated as a syrup 111 mg, quant ; NMR
Ž
.
Ž
.
1
Ž
.
Ž
.
Ž
.
Ž
CDCl3 : H, d 7.42–7.08 m, 20 H, 4 Ph , 6.904 d,
1
13
Ž
.
Ž
X
X
.
Ž
.
1 H, JP – H 614 Hz, P–H ; C, d 78.6 C-2,3,4 ,
73.6, 73.1, 72.3, 72.0, 70.3, and 63.0 4 OCH2 Ph and
X
X
.
Ž
.
X
X
X
X
Ž
Ž
Ž
.
.
.
Ž
.
w
Ž
.
x
w
Ž
.
x
.
C-1,5 , 45.1 N CH2CH3 3 , 8.0 N CH2CH3 3
Ž
X
X
(
3-N-Benzyloxycarbonylaminopropyl 1,2,3,4-tetra-
X
13
X
.
Ž
.
Ž
.
. Ž
O-benzyl-D-ribityl - 5 ™ triethylammonium phos-
Ž
.
Ž
)
73.4 2 C , 72.8, 71.7, 71.6, and 70.6 C-
2,3,2 ,3 ,4 ,5 , 65.8 and 64.7 OCH2 CH2 2 N and
phate™2 -3,4,6-tri-O-benzyl-a-D-galactopyrano-side
X
X
X
X
.
Ž
Ž
.
Ž
.
Ž
42 and 3-aminopropyl D-ribitol- 5 ™ hydrogen
.
Ž
.
.
Ž
.
Ž .
C-5, JP,OC-spacer f JP,C-5 f 5.4 Hz , 64.0 C-1 , 38.6
phosphate™2 - a - D - galactopyranoside 43 .— a
w
Ž
.
x
Ž
OCH 2 C H 2 CH 2 N,
Ž
.
Ž
O CH 2 2 C H 2 N , 29.2
A mixture of 8 46 mg, 89 mmol and 39 97 mg,
31
q
.
Ž
.
.
Ž
.
JP,OC
6.8 Hz ; P, d 3.71 PO4 . FABMS
0.12 mmol in 1:5 pyridine–acetonitrile 2.6 mL was
stirred for 10 min, and pivaloyl chloride 70 mL, 0.57
mmol was added. After 17 h, an additional amount
of pivaloyl chloride 50 mL, 0.41 mmol was added.
After 24 h TLC 9:1 CH2Cl2–acetone revealed the
disappearance of 8 and the appearance of a new spot
with Rf 0.60 41 . A 0.5 M solution of iodine in 95:5
pyridine–water 360 mL was added. After 18 h,
C -spacer
w
xq
Ž
Calcd for C14H30NO12 P: mrz 436.4 MqH .
Found: mrz 436.4 MqH .
w
xq
.
Ž
.
3-N-Benzyloxycarbonylaminopropyl 3,4,6-tri-O-
Ž
.
Ž
.
benzyl-2-O- triethylammonium H-phosphonate -a-
D-galactopyranoside 39 .—To a soln of 12 75 mg,
117 mmol in 1:5 pyridine–acetonitrile 6 mL was
added 2-chloro-4H-1.3.2-benzodioxaphosphorin-4-
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
Ž
.
one 32 mg, 158 mmol , and the mixture was stirred
TLC 9:1 CH2Cl2–acetone showed the disappear-

(
)
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
107
ance of 41 and a new spot on the baseline. The
mixture was diluted with CH2Cl2, washed with aq
5% Na2S2O3 and 1 M triethylammonium bicarbonate
disappearance of 5 and the appearance of a new spot
Ž
.
with Rf 0.47 44 . A 0.5 M solution of iodine in 95:5
Ž
.
pyridine–water 50 mL was added, and after 3 h
Ž
.
Ž
.
Ž
.
2= , dried MgSO4 , filtered, and concentrated.
TLC 9:1 CH2Cl2–acetone indicated the disappear-
ance of 44. The mixture was diluted with CH2Cl2,
washed with aq 5% Na2S2O3 and 1 M triethylammo-
Ž
Column chromatography 80:19:1 CH2Cl2–acetone–
Et3N, then 80:19:1 CH2Cl2–MeOH–Et3N of the
residue gave 42, isolated as a glass 45 mg, 38% . b
A mixture of 40 145 mg, 0.214 mmol and 12 106
mg, 0.165 mmol in 1:5 pyridine–acetonitrile 5.4
mL was stirred for 10 min, and pivaloyl chloride
Ž
additional amount of pivaloyl chloride 100 mL, 0.813
mmol was added. After 20 h, TLC 9:1 CH2Cl2–
acetone revealed the disappearance of 12. A 0.5 M
solution of iodine in 95:5 pyridine–water 350 mL
.
Ž
. Ž .
Ž
.
Ž
.
nium bicarbonate 2= , dried MgSO4 , filtered, and
Ž
.
.
Ž
Ž
concentrated. Column chromatography 80:19:1
Ž
toluene–acetone–Et3N, then 80:19:1 toluene–
.
.
MeOH–Et3N of the residue gave 45, isolated as a
1
.
Ž
.
Ž
.
100 mL, 0.813 mmol was added. After 18 h, an
glass 21 mg, 89% ; NMR CDCl3 : H, d 7.95–7.45
m, 35 H, 7 Ph , 2.722 q, 6 H, N CH2CH3 3 ,
2.020, 1.947, and 1.940 3 s, each 3 H, 3 Ac , 1.112
t, 9 H, N CH2CH3 3 , 0.907 d, 3 H, J5 ,6 6.2 Hz,
3 H-6 ; C, d 169.8 COCH3 , 166.3 COPh ,
156.7 NCOOCH2 Ph , 98.7 and 95.8 C-1,1 , 45.2
Ž
Ž
.
Ž
Ž
. .
.
Ž
Ž
.
Y
Y
.
Ž
Ž
. .
Ž
Y
13
Ž
.
.
.
Ž
.
Ž
.
Y
Ž
Ž
.
Ž
.
was added. After 20 h, TLC 9:1 CH2Cl2–acetone
w
Ž
Ž
.
x
w Ž . x
37.5 O CH 2 2C H 2 N , 29.5
indicated the oxidation to be completed, and the
mixture was diluted with CH2Cl2, washed with aq
5% Na2S2O3 and 1 M triethylammonium bicarbonate
N C H 2CH 3
,
3
.
Ž
Ž
31
. Ž
.
OCH2CH2CH2 N , 20.7 and 20.6 2 C 3 COCH3 ,
17.0 C-6 , 8.3 N CH2CH3 3
Y
Ž
.
w
Ž
.
x
;
Ž .
P, d y2.57 PO4 .
Ž
.
Ž
.
.
2= , dried MgSO4 , filtered, and concentrated.
A soln of 45 21 mg, 14 mmol in 2:1 MeOH–aq
Ž
Ž
.
Column chromatography 80:19:1 CH2Cl2–acetone–
Et3N, then 80:19:1 CH2Cl2–MeOH–Et3N of the
25% NH4OH 5 mL was heated for 24 h at 50 8C,
then concentrated. This de-O-acylation procedure was
repeated twice, to yield crude 46, which was purified
.
Ž
.
residue gave 42, isolated as a syrup 88 mg, 40% ;
1
Ž
.
Ž
.
Ž
H NMR CDCl3 : d 7.31–7.15 m, 40 H, 8 Ph ,
on Sephadex LH-20 50:50:1 CH2Cl2–MeOH–
Ž
.
Ž
.
5.265 d, 1 H, J1,2 3.8 Hz, H-1 , 5.062 and 4.979 2
d, each 1 H, COOCH2 Ph , 4.840, 4.829, 4.711,
Et3N . To a soln of 46 in 1:2:2:2 water–EtOAc–2-
propanol–EtOH 5 mL was added 10% Pd–C 10
mg , and the mixture was hydrogenolysed at atmo-
.
Ž
.
Ž
.
4.665, 4.634, 4.584, 4.523, 4.480, 4.449, and 4.374
Ž
.
Ž
10 d, each 1 H, 5 OCH2 Ph , 4.633 and 4.421 2 d,
spheric pressure for 16 h, filtered, and concentrated.
The hydrogenolysis procedure was repeated twice,
and the crude product was purified by Bio-Gel P-2
gel-permeation chromatography using water as elu-
.
Ž
Ž
. .
each 2 H, 2 OCH2 Ph , 2.952 q, 6 H, N CH2CH3 3 ,
1.237 t, 9 H, N CH2CH3 3 .
Ž
Ž
. .
Ž
.
.
To a soln of 42 120 mg, 91.1 mmol in 1:2:2:2
Ž
Ž
water–EtOAc–2-propanol–EtOH 7.5 mL was added
ent, affording 47, isolated as a white powder 3.9 mg,
1
Ž
.
.
Ž
.
Ž
10% Pd–C 20 mg , and the mixture was hy-
drogenolysed at 4 kgrcm2 for 24 h. After filtration,
the mixture was concentrated, and the residue was
purified by Bio-Gel P-2 gel-permeation chromatogra-
47% ; H NMR D2O : d 5.164 d, 1 H, J1,2 4.0 Hz,
H-1 , 5.086 d, 1 H, J1 ,2 1.7 Hz, H-1 , 3.462 t, 1
Y
.
Ž
.
Ž
Y
Y
Y
.
Ž
Y
Y
Y
Y
H, J3 ,4 sJ4 ,5 s9.7 Hz, H-4 , 3.22–3.11 m, 2 H,
Ž
.
.
Ž
O C H 2 C H 2 N , 2.04 – 1.98
m ,
Y
2
H ,
2
.
Ž
Y
phy using water as eluent, affording 43, isolated as a
OCH2CH2CH2 N , 1.292 d, 3 H, J5 ,6 6.3 Hz, 3
1
H-6 ; 31P, d 0.65 PO4 . FABMS Calcd for
Y
q
Ž
.
Ž
.
.
Ž
.
white powder 32 mg, 77% ; H NMR D2O : d
w
xq
C20H40NO17P: mrz 598.5 MqH . Found: mrz
Ž
.
Ž
5.178 d, 1 H, J1,2 3.8 Hz, H-1 , 3.28–3.11 m, 2 H,
O C H 2 C H 2 N , 2.07 – 1.96
OCH2C H2CH2 N , 1.331 d, 3 H, J5,6 6.5 Hz, 3
w
xq
Ž
.
.
Ž
m ,
2
H ,
598.5 MqH .
2
.
Ž
(
3-N-Benzyloxycarbonylaminopropyl 1,2,3,4-tetra-
q
.
) (
H-6 . FABMS Calcd for C14H30NO13P: mrz 452.4
O - benzyl - D - ribityl - 5™triethylammonium phos-
w
xq
w
xq
) (
)
.
MqH . Found: mrz 452.4 MqH .
phate™2 - 3,4,6-tri-O-acetyl-a-D-galactopyranosyl -
(
(
)
Ž
3-N-Benzyloxycarbonylaminopropyl 2,3,4-tri-O-
1™3 -2,4,6-tri-O-benzyl-a-D-glucopyranoside 49
. Ž
. Ž
(
acetyl-a-D-rhamnopyranosyl - 1™4 - 1-O-benzeyl-
and 3 - aminopropyl D - ribityl- 5™hydrogen phos-
.
Ž
)
(
)
2,3 - di - O - benzyl - D - ribityl
-
5™
phate™2 - a - D - galactopyranosyl - 1™3 - a - D -
)
Ž
.
Ž
triethylammonium phosphate ™ 2 -3,4,6-tri-O-
glucopyranoside 51 .—A mixture of 40 37 mg, 55
Ž
.
.
Ž
.
benzyl-a-D-galactopyranoside 45 .—A mixture of
mmol and 33 54 mg, 58 mmol in 1:5 pyridine–
Ž
.
Ž
.
Ž
.
39 19 mg, 24 mmol and 5 11 mg, 16 mmol in 1:5
acetonitrile 2.5 mL was stirred for 15 min, and
Ž
Ž
.
Ž
.
pyridine–acetonitrile 1.6 mL was stirred for 10 min,
pivaloyl chloride 30 mL, 0.25 mmol was added.
.
and pivaloyl chloride 20 mL, 0.16 mmol was added.
After 3 h, TLC 9:1 CH2Cl2–acetone revealed the
After 5 h, an additional amount of pivaloyl chloride
Ž
.
Ž
.
30 mL, 0.25 mmol was added, and after 18 h TLC

(
)
108
M.J.L. Thijssen et al.rCarbohydrate Research 306 1998 93–109
Ž
.
9:1 CH2Cl2–acetone revealed the disappearance of
References
Ž
.
33 and the formation of a new spot 48 . A 0.5 M
soln of iodine in 95:5 pyridine–water 82 mL was
added, and after 4 h TLC 8:2 CH2Cl2–acetone
showed the disappearance of 48. The mixture was
diluted with CH2Cl2, washed with aq 5% Na2S2O3
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.
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.
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MgSO4 , filtered, and concentrated. Column chro-
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.
80:19:1 CH2Cl2–MeOH–Et3N of the residue gave
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.
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.
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.
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.
Ž
CDCl3 : d 7.33–7.16 m, 40 H, 8 Ph , 5.872 d, 1
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.
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.
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.
X
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H-2 , 5.25 d, 1 H, J3 ,4 -1 Hz, H-3 , 5.072 and
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J1,2 3.4 Hz, H-1 , 4.230 dd, 1 H, J2,3 9.4, J3,4 9.0
Hz, H-3 , 3.424 dd, 1 H, H-2 , 2.947 q, 6 H,
N CH2CH3 3 , 1.928 and 1.901 2 s, 3,6 H, 3 Ac ,
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.
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.
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.
A soln of 49 42 mg, 26 mmol in 2:1 MeOH–aq
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¨
¨
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.
25% NH4OH 7.5 mL was heated for 24 h at 50 8C,
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1 H, J1 ,2 2.5 Hz, H-1 , 4.944 d, 1 H, J1,2 2.4 Hz,
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.
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