Synthesis of lipid A type pyran carboxylic acids with ether chains and their biological activities

Article abstract of DOI:10.1016/S0008-6215(01)00134-3

Synthesis of lipid A type pyran carboxylic acids having ether chains at both the C-3′ and C-4 positions and their bioactivities toward human U937 cells are described.

Full text of DOI:10.1016/S0008-6215(01)00134-3

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Carbohydrate Research 333 (2001) 203–231
Synthesis of lipid A type pyran carboxylic acids with
ether chains and their biological activities
Yukiko Watanabe,a Takashi Mochizuki,a Masao Shiozaki,a,* Saori Kanai,b
Shin-ichi Kurakata,b Masahiro Nishijimac
aExploratory Chemistry Research Laboratories, Sankyo Co. Ltd., Hiromachi 1-2-58, Shinagawa-ku,
Tokyo 140-8710, Japan
bBiological Research Laboratories, Sankyo Co. Ltd., Hiromachi 1-2-58, Shinagawa-ku, Tokyo 140-8710, Japan
cDepartment of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Toyama 1-23-1,
Shinjuku-ku, Tokyo 162-8640, Japan
Received 15 March 2001; accepted 30 April 2001
Abstract
Synthesis of lipid A type pyran carboxylic acids having ether chains at both the C-3% and C-4 positions and their
bioactivities toward human U937 cells are described. © 2001 Elsevier Science Ltd. All rights reserved.
Keywords: Lipid A; Endotoxin; LPS-agonist; LPS-antagonist; TNFa production inhibitor
1. Introduction
polysaccharide chain to the cell wall. Ana-
logues of both the nonreducing distal subunit
and the reducing sugar part of lipid A are
usually still biologically active.3
Endotoxin (lipopolysaccharide; LPS)1 is a
toxic substance of Gram-negative bacteria and
one of the components of their outer surface
membrane. It is also a highly potent stimula-
tor of the immune system.2 A variety of re-
sponses, both beneficial and harmful, can be
elicited by LPS. One of these harmful re-
sponses is fatal endotoxic shock (bacterial sep-
sis) caused as a consequence of acute
inflammatory response, and this fatal shock
has precluded the clinical use of LPS. Most of
the biological activities of LPS reside in a
relatively small portion of the molecule, that
is, the terminal disaccharide phospholipid sub-
unit known as lipid A, which is a hydrophobic
anchor substance holding an essentially linear
LPS and its related compounds have mainly
been investigated as anticancer drugs4 that
function as LPS-agonists by activating
macrophages. However, in recent years, lipid
A-related compounds have been studied as
LPS-antagonists, which may have potential as
immunosuppressants,5 and in autoimmune
diseases5 and septicemia6 by deactivating LPS-
induced aggressive macrophages. For exam-
ple, Qureshi’s group7 isolated a nontoxic lipid
A-related compound from Rhodobacter
sphaeroides as an LPS antagonist, and an Ei-
sai group recently developed a related com-
pound, E5564,6,8 as
anti-septicemia drug.
a
highly potent
On the other hand, during our investigation
of the biological activities of compounds re-
lated to GLA-60,3a which is a nonreducing
* Corresponding author. Tel.: +81-3-34913131; fax: +81-
3-54368570.
E-mail address: shioza@shina.sankyo.co.jp (M. Shiozaki).
0008-6215/01/$ - see front matter © 2001 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(01)00134-3

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Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
distal subunit analogue in the lipid A
molecule, we also found that most of them
had LPS-agonistic activity, but a few of them
behaved as LPS antagonists. The anomeric
carboxymethyl GLA-60 analogue A9 and the
pyran carboxylic acid B10 exhibited fairly
strong LPS-antagonistic activity, and also
lipid A-type disaccharides constructed from
pyran carboxylic acid C11 showed a strong
LPS-antagonistic activity toward human U937
cells. Compound C has an acetamido group in
the 2%-position, four fatty acid chains, and a
carboxylic acid at the anomeric position.
Therefore, by using this information on the
characteristics of C, we synthesized the title
compounds which have both C-4–O and C-
3%–O ether bonds instead of ester bonds in
compound C, in order to stabilize the com-
pound and to increase its activity. Herein we
report the synthesis of lipid A-type pyran
carboxylic acid analogues with ether chains
and their activities (Fig. 1).
reduction12 in THF of commercially available
(R)-3-hydroxytetradecanoic acid, was treated
with 4-methoxybenzaldehyde dimethylacetal
using p-TsOH·H2O as a catalyst to yield single
acetal 3. The configurations of two sub-
stituents of compounds 3 should be equatorial
toward the [1,3]dioxane ring, because the
equatorial configuration should be more sta-
ble than axial. DIBAL reduction of com-
pound 3 in CH2Cl2 gave 4 exclusively without
detection of a secondary alcohol. Mesylation
of 4 with methanesulfonyl chloride using Et3N
as a base yielded 5.
Pyran carboxylic acid 113 as a glycosyl ac-
ceptor should be coupled with the glycosyl
donors, the imidates obtained from com-
pounds 15, 22, 24, 54, 59 and 61, to yield the
pseudo-disaccharides 28, 29, 30, 62, 63 and 64,
respectively (Schemes 5 and 9). Therefore, we
converted trifluoroacetamide 614 to com-
pounds 15, 22, 24, 54, 59 and 61 as shown in
Schemes 2–4, 7 and 8.
The glycosyl donor precursor 15 was syn-
thesized from trifluoroacetamide 6 as follows
(Scheme 2). The C-3 hydroxy compound 6
was alkylated with (R)-3-(4-methoxybenzyl)-1-
(methanesulfonyloxy)tetradecane and sodium
hydride in N,N-dimethylformamide (DMF) at
room temperature to give 7. The trifluoroac-
etamide of 7 was hydrolyzed with aqueous 1
2. Results and discussion
Synthesis.—First, in Scheme 1, mesylates 5
as side-chain material was prepared from opti-
cally active starting diol 2. The starting diol 2,
easily obtained by lithium aluminum hydride
Fig. 1. LPS-antagonists towards human U937 cells.
Scheme 1. Reagents and conditions: PMB=p-methoxybenzyl; (a) 4-methoxybenzaldehyde dimethylacetal, p-TsOH, DMF, 99%;
(b) DIBAL, CH2Cl2, −78 °C, 93%; (c) MsCl, Et3N, CH2Cl2, 0 °C, 99%.

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
205
Scheme 2. Reagents and conditions: (a) (R)-3-(4-methoxybenzyloxy)-1-(methanesulfonyloxy) tetradecane, NaH, DMF, rt, 18 h,
84%; (b) aq 1 M NaOH, EtOH, 60 °C, 3 h, 99%; (c) ClCOOCH2CCl3, satd aq NaHCO3, CH2Cl2, rt, 30 min, 94%; (d) 80% aq
AcOH, 60 °C, 1 h, 83%; (e) ClCOOBn, pyridine, CH2Cl2, rt, 2 h, 97%; (f) (PhO)2P(O)Cl, DMAP, CH2Cl2, rt, 1 h, 94%; (g) DDQ,
CH2Cl2–water, rt, 1 h, 94%; (h) C13H27COCl, Et3N, THF, rt, 18 h, 93%; (i) [C8H12Ir(PMePh2)2] PF6, THF, rt, 3 h; then THF,
water, pyridine, I2, rt, 1 h, 81%.
in tetrahydrofuran (THF) yielded ester 14.
Double bond migration of the anomeric allyl
group of 14 to anomeric vinyl ether by treat-
ment with (1,5-cyclooctadiene)bis(methyl-
diphenylphosphine)iridium(I) hexafluorophos-
phate, and successive oxidative hydrolysis
with water–pyridine–iodine in THF afforded
cyclic hemiacetal 15 in 65% overall yield from
10 in five steps.
M NaOH in EtOH to convert to amine 8.
Treatment of 8 with 2,2,2-trichloroethyl chlo-
roformate and satd aqueous NaHCO3 as a
base in dichloromethane gave carbamate 9.
The O-isopropylidene group of 9 was re-
moved by treatment with aqueous 80% AcOH
at 60 °C for 3 h to give diol 10. The overall
yield of 10 from 6 was 65% in four steps.
Treatment of 10 with benzyl chloroformate
and pyridine in dichloromethane gave carbon-
ate 11, regioselectively. Phosphorylation of the
C-4 alcohol of 11 with diphenyl chlorophos-
phate and 4-dimethylaminopyridine (DMAP)
yielded phosphate 12 without formation of
cyclic carbonate in those two steps from 10.
Deprotection of the 4-methoxybenzyl group
on the alkyl side chain of 12 by 2,3-dichloro-
5,6-dicyano-1,4-benzoquinone (DDQ) gave al-
cohol 13. Acylation of the alcohol group of 13
with tetradecanoyl chloride and triethylamine
The glycosyl donor precursor 22,
a
methoxylated compound at the C-6 position,
was synthesized from diol 10 as follows
(Scheme 3). Regioselective silylation of the
primary C-6–OH group of 10 with tert-
butyldimethylsilyl chloride (TBDMSCl) and
DMAP as a base gave silyl ether 16. Com-
pound 16 was converted to 19 via 17 and 18
by the same procedure as in the formation of
14 from 11. Desilylation of 19 was performed
by treatment with aqueous 3 M HCl in THF

206
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
etal 24 according to the same deallylation
procedure from 14 to 15. The yield of 24 from
20 was 58% in two steps.
to give alcohol 20 in 74% overall yield from 10
in five steps. Methylation of the C-6–OH
group of 20 using trimethyloxonium tetra-
fluoroborate and 2,6-di-tert-butyl-4-methyl-
pyridine as a base gave C-6–OMe ether 21,
which was further converted to cyclic hemiac-
etal 22 according to the same deallylation
procedure from 14 to 15. The yield of 22 from
20 was 66% in two steps.
The glycosyl donor precursor 24, a fluori-
nated compound at the C-6 position, was
synthesized from the primary alcohol 20 as
follows (Scheme 4). Fluorination of the C-6–
OH group of 20 using diethylaminosulfur trifl-
uoride (DAST) gave C-6–F compound 23,
which was further converted to cyclic hemiac-
The coupling of 15, 22, and 24 via the
corresponding imidates 25, 26, and 27 with 1,
and successive conversion to corresponding
2%-acetamides 37, 38, and 39 are shown in
Scheme 5. Treatment of 15, 22, and 24 with
trichloroacetonitrile
using
1,8-diazabicy-
clo[5.4.0]undec-7-ene (DBU) as a base gave
imidates 25, 26, and 27, which were indepen-
dently condensed with
1
using both
trimethylsilyl trifluoromethanesulfonate (TM-
,
SOTf) and 4 A molecular sieves (MS4A) as
condensing reagents to afford 28, 29, and 30,
respectively. Each trichloroethoxycarbonyl
Scheme 3. Reagents and conditions: (a) TBDMSCl, DMAP, CH2Cl2, rt, 3 h, 96%; (b) (PhO)2POCl, DMAP, CH2Cl2, rt, 1 h, 98%;
(c) DDQ, CH2Cl2–water, rt, 1 h, 91%; (d) C13H27COCl, Et3N, DMAP, THF, rt, 24 h, 95%; (e) 3 M HCl, THF, rt, 2 h, 91%; (f)
Me3OBF4, 2,6-di-tert-butyl-4-methylpyridine, CH2Cl2, rt, 3 h, 91%; (g) [C8H12Ir(PMePh2)2]PF6, THF, rt, 2 h; then THF, water,
pyridine, I2, rt, 2 h, 73%.
Scheme 4. Reagents and conditions: (a) DAST, DME, 0 °C, 3 h, 80%; (b) [C8H12Ir(PMePh2)2]PF6, THF, rt, 3 h; then water,
pyridine, I2, THF, 2 h, 73%.

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
207
Scheme 5. Reagents and conditions: (a) (i) Cl3CCN, cat. DBU, CH2Cl2, 0 °C, 1 h; (ii) 1, cat. TMSOTf, MS4A, CH2Cl2, −78 °C,
2 h, 28 (88%): 29 (72%): 30 (72%); (b) (i) Zn, AcOH, rt, 24 h; (ii) Ac2O, pyridine, THF–water, rt, 4 h, 31 (70%): 32 (65%): 33
(66%); (c) H2, 20% Pd(OH)2–C, EtOH, rt, 14 h, 34 (52%): 35 (72%): 36 (64%); (d) H2, PtO2, THF, rt, 18 h, 37 (95%): 38 (96%):
39 (98%).
46, 47, and 48 via 43, 44, and 45 according to
the same deprotection procedure in the forma-
tion of 37, 38, and 39 from 31, 32, and 33,
respectively.
(Troc) group as an amine protecting group of
28, 29, and 30 was removed with zinc–acetic
acid, and successive treatment of each corre-
sponding amine with acetic anhydride–pyri-
dine yielded acetamides 31, 32, and 33. The
protecting groups for hydroxy and ester
groups of 31, 32, and 33 were removed by
hydrogenolysis using palladium hydroxide-on-
charcoal to give 34, 35, and 36, which were
successively deprotected to 37, 38, and 39 by
hydrogenolysis using platinum oxide as a
catalyst.
Compounds 46, 47, and 48 having a [(R)-3-
dodecanoyloxytetradecanamido] group at the
C-2%-amide position were synthesized from 28,
29, and 30, respectively, as follows (Scheme 6).
The amines derived from 28, 29, and 30 were
treated with 3-(R)-3-dodecanoyloxytetra-
decanoic acid and dicyclohexylcarbodiimide
(DCC) to give amides 40, 41, and 42, respec-
tively. These amides were further converted to
Next, (R)-3%-O-dodecyloxytetradecyl ana-
logues 71, 72, and 73 were synthesized as
follows (Schemes 7–9). The glycosyl donor
precursor 54 was synthesized from trifluoroac-
etamide 614 according to almost the same pro-
cedures as in the formation of 15 from 6 as
follows (Scheme 7). C-3–OH compound 6 was
alkylated with (R)-3-(dodecyloxy)-1-(methane-
sulfonyloxy)tetradecane14 and NaH in DMF
at room temperature to give C-3 ether 49. The
trifluoroacetamide of 49 was hydrolyzed with
aqueous 1 M NaOH in EtOH to convert it to
the corresponding amine, which was treated
with 2,2,2-trichloroethyl chloroformate and
satd aqueous NaHCO3 as
a
base in
dichloromethane to give carbamate 50. The
O-isopropylidene group of 50 was deprotected

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Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
ment with (1,5-cyclooctadiene)bis(methyl-
diphenylphosphine)iridium(I) hexafluorophos-
phate, and successive oxidative hydrolysis
with water–pyridine–iodine in THF afforded
cyclic hemiacetal 54 in 64% yield from 51.
Additionally, C-6-methoxy and C-6-fluoro
compounds 59 and 61 as glycosyl donors were
also synthesized from diol 51 according to
almost the same procedures as in the forma-
by treatment with aqueous 80% AcOH at
60 °C for 3 h to give diol 51 in 62% from 6.
Treatment of 51 with benzyl chloroformate
and pyridine in CH2Cl2 gave carbonate 52
regioselectively. Phosphorylation of the C-4–
OH group of 52 with diphenyl chlorophos-
phate and DMAP yielded phosphate 53.
Double bond migration of the anomeric allyl
group of 53 to anomeric vinyl ether by treat-
Scheme 6. Reagents and conditions: (a) (i) Zn, AcOH, rt, 20 h; (ii) (R)-3-(dodecanoyloxy)tetradecanoic acid, DCC, (ClCH2)2, rt,
6 h, 40 (54%), 41 (71%), and 42 (58%), respectively; (b) H2, 20% Pd(OH)2–C, EtOH, rt, 14 h, 43 (49%), 44 (77%), and 45 (58%),
respectively; (c) H2, PtO2, THF, rt, 18 h, 46 (98%), 47 (77%), and 48 (88%), respectively.
Scheme 7. Reagents and conditions: (a) (R)-3-(dodecyloxy)-1-(methanesulfonyloxy)tetradecane, NaH, DMF, rt, 3 h, 75%; (b) 1 M
aq NaOH, EtOH, 60 °C, 4 h; then ClCOOCH2CCl3, satd aq NaHCO3, CH2Cl2, rt, 1 h, 87%; (c) 80% aq AcOH 60 °C, 4 h, 95%;
(d) ClCOOBn, pyridine, CH2Cl2, rt, 3 h, 94%; (e) (PhO)2P(O)Cl, DMAP, CH2Cl2, rt, 2 h, 96%; (f) [C8H12Ir(PMePh2)2]PF6, THF,
rt, 3 h; then water, pyridine, I2, THF, rt, 1 h, 71%.

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
209
Scheme 8. Reagents and conditions: (a) TBDMSCl, DMAP, CH2Cl2, rt, 3 h, 99%; (b) (PhO)2P(O)Cl, DMAP, CH2Cl2, rt, 2 h,
99%; (c) 3 M HCl, THF, rt, 3 h, 89%; (d) Me3OBF4, 2,6-di-tert-butyl-4-methylpyridine, CH2Cl2, rt, 3 h, 91%; (e)
[C8H12Ir(PMePh2)2]PF6, THF, rt, 3 h; then water, pyridine, I2, THF, rt, 1 h, 72% (59 from 58) and 82% (61 from 60), respectively;
(f) DAST, DME, −40 °C, 30 min, then rt, 2 h, 83%.
tion of 22 and 24 from 10 as follows (Scheme
8). The diol 51 was selectively protected as
silyl ether 55 using TBDMSCl and DMAP
and was further converted to phosphate 56
using diphenyl chlorophosphate and DMAP.
Treatment of 56 with aqueous 3 M HCl in
THF afforded alcohol 57, which was con-
verted to C-6–OMe compound 58 or C-6–F
compound 60 by treatment with Me3OBF4 or
DAST. Anomeric allyl group deprotection of
58 and 60 according to the same procedure as
in the formation of 15 from 14 was performed
to give cyclic hemiacetals 59 and 61, respec-
tively. The yields of 59 and 61 from 51 were
57 and 59%, respectively.
Pseudodisaccharides 71, 72, and 73 were
synthesized from 54, 59 and 61, respectively,
by coupling reaction with 1, and by a series of
conversions of corresponding disaccharides
according to almost the same procedures as in
the formation of 37, 38 and 39 from 15, 22
and 24 (Scheme 9). Activation of the anomeric
hydroxy group of 54, 59 and 61 with
trichloroacetonitrile using catalytic amount of
cesium carbonate afforded corresponding
trichloroacetimidate compounds, which were
immediately used for subsequent glycosylation
with diol 1 in the presence of TMSOTf and
MS4A to give pseudodisaccharides 62, 63 and
64, respectively. Deprotection of the C-2%–N-
Troc groups of 62, 63 and 64 with zinc dust in
acetic acid, and successive acetylation of the
corresponding amines with acetic anhydride
and pyridine gave acetamides 65, 66 and 67,
respectively. Treatment of 65, 66 and 67 in
hydrogen using 20% palladium hydroxide-on-
charcoal as a catalyst gave carboxylic acids
68, 69 and 70, respectively. Finally, hy-
drogenolysis of 68, 69 and 70 using PtO2 as a
catalyst gave phosphoric acids 71, 72 and 73,
respectively. The yields of 71, 72 and 73 from
54, 59 and 61 were 37, 45, and 41%,
respectively.
Biological acti6ity.—The inhibitory activity
of compounds 37, 38, 39, 46, 47, 48, 71, 72
and 73 on LPS-induced TNFa production was
investigated in vitro using human monoblastic
U937 cells. Compounds 37, 38, 39, 71, 72 and
73, which have four chains in their molecules,
inhibited TNFa production as LPS-antago-
nists, and the IC50 (nM) values of these six
compounds were 0.86, 0.61, 1.2, 11, 6.4 and
10, respectively. Compound 47, having six
chains in the molecule, was almost inactive.

210
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
Scheme 9. Reagents and conditions: (a) Cl3CCN, cat. Cs2CO3, CH2Cl2, rt, 1 h; then 1, cat. TMSOTf, MS4A, CH2Cl2, −40 °C,
2 h, 75% (62), 68% (63) and 80% (64), respectively; (b) Zn, AcOH, rt, 5 h; then Ac2O, pyridine, THF–water, rt, 2 h, 71% (65),
74% (66) and 68% (67), respectively; (c) H2, 20% Pd(OH)2–C, EtOH, rt, 18 h, 72% (68), 89% (69) and 80% (70), respectively; (d)
H2, PtO2, THF, rt, 5 h, 96% (71), 100% (72) and 94% (73), respectively.
3. Experimental
On the other hand, compounds 46 and 48
having six chains in the molecules, showed
moderate acceleration of TNFa production as
LPS-agonists as shown in Figs. 2 and 3.
Judging from the activity of these com-
pounds, the total number of chains in the
molecules plays an important role in whether
or not the compound exhibits LPS-antago-
nism or LPS-agonism. If the total number of
chains is four, the compounds are LPS-antag-
onists. However, if it is six, the compounds
are LPS-agonists or inactive. And, also the
three C-6 substitution patterns having hy-
droxy, methoxy and fluorine groups were in-
vestigated. However, there were no big
differences between them in their activity. This
tendency is correlated with the previous re-
sults.11 The difference between 3-tetra-
decanoyloxytetradecyl and 3-dodecyloxy-
tetradecyl in the C-3%–O substituent is not
critical for inhibitory activity toward human
monoblastic U937 cells.
Melting points are uncorrected. Optical ro-
tations were obtained by the use of a JASCO
P-1030 polarimeter. IR absorption spectra
were recorded on a JASCO IR A-2 spec-
trophotometer. 1H NMR spectra were
recorded with a JEOL-GSX 400 spectrometer
using Me4Si as an internal standard, and mass
Fig. 2. Inhibitions of TNFa release from TPA-treated U937
cells stimulated by LPS (30 ng/mL) in the presence of lipid
A-type pyrancarboxylic acid derivatives.

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
211
3.80), 3.94 (1 H, m), 4.24 (1 H, m), 5.46 (1 H,
s), 6.89 (2 H, d, J 8.6 Hz), 7.42 (2 H, d, J 8.7
Hz). FABMS (positive-ion): m/z 349 [M+
H]+. HRFABMS (positive-ion): Calcd for
C22H37O3: 349.2743; Found: 349.2751. Anal.
Calcd for C22H36O3 (348.5): C, 75.82; H,
10.41. Found: C, 75.60; H, 10.68.
(ii) To a solution of the above-obtained 3
(2.71 g, 7.80 mmol) in CH2Cl2 (25 mL) was
added DIBAL (1.0 M solution in CH2Cl2, 24
mL, 24.0 mmol) at −78 °C under N2. After
stirring for 2 h at 0 °C, the reaction mixture
was quenched with satd NH4Cl at 0 °C, and
0.5 M aq Rochelle salt was added to this
mixture. After stirring for 30 min at rt, the
mixture was extracted with CH2Cl2, washed
with water and brine, dried over MgSO4,
filtered, and concentrated in vacuo to give a
mixture that was chromatographed on a silica-
gel column. Elution with 7:3 cyclohexane–
EtOAc gave (R)-1-hydroxy-3-(4-methoxy-
benzyloxy)tetradecane (4, 2.54 g, 93%) as an
oil. IR wmax(CHCl3) 3626, 3499, 2929, 2857,
Fig. 3. Inhibitions of TNFa release from U937 cells from
TPA-treated U937 cells stimulated by LPS (30ng/mL) in the
presence of lipid A-type pyrancarboxylic acid derivatives.
spectra were obtained with a JMS-700 mass
spectrometer. Elemental analyses were per-
formed by the Institute of Science and Tech-
nology, Inc. Separation of the compounds by
column chromatography was carried out with
Silica Gel 60 (E. Merck, 0.040–0.063 mm) at
slightly elevated pressure (1.1–1.5 atm) for
easy elution, and the quantity of the used
silica gel was 50–100 times the weight of the
purified compounds. Analytical chromatogra-
phy was performed on E. Merck Art 5715
Silica Gel 60-F254 plates. THF was distilled in
the presence of radical anions generated by
Na–benzophenone ketyl. Dichloromethane
was dried by being passed through an ICN
Alumina B-Super I, and DMF and pyridine
1
1613 cm−1. H NMR (270 MHz, CDCl3) l
0.88 (3 H, t, J 6.6 Hz), 1.27–1.84 (22 H, m),
2.44 (1 H, t, J 5.4 Hz, OH), 3.62 (1 H, m),
3.69–3.82 (5 H, m, containing 3 H, s, at l
3.80), 4.41, 4.54 (2 H, AB-q, J 11.1 Hz), 6.88
(2 H, d, J 8.6 Hz), 7.26 (2 H, d, J 8.6 Hz).
FABMS (positive-ion): m/z 373 [M+Na]+.
HRFABMS (positive-ion): m/z Calcd for
C22H38O3: 350.2821; Found: 350.2800. Anal.
Calcd for C22H38O3 (350.5): C, 74.77; H,
11.10. Found: C, 75.08; H, 10.93.
,
were dried by storage over 4 A molecular
sieves.
(R)-1-Methanesulfonyloxy-3-(4-methoxy-
benzyloxy)tetradecane (5).—(i) To a solution
of (R)-1,3-dihydroxytetradecane (2, 1.85 g, 8.0
mmol) in DMF (20 mL) were added 4-
methoxybenzaldehyde dimethylacetal (2.90 g,
16.0 mmol) and p-TsOH·H2O (152 mg, 0.80
mmol). After stirring at rt for 2 h, the reaction
mixture was diluted with EtOAc, washed with
satd NaHCO3 and brine, dried over MgSO4,
filtered, and concentrated in vacuo to give a
mixture, which was chromatographed on a
silica-gel column. Elution with 9:1 cyclohex-
ane–EtOAc gave 2-(4-methoxyphenyl)-4-(R)-
undecyl-[1,3]dioxane (3, 2.78 g, 99%) as an oil.
(iii) To a solution of 4 (2.50 g, 7.10 mmol)
in CH2Cl2 (20 mL) were added Et3N (1.5 mL,
10.7 mmol) and methanesulfonyl chloride
(1.22 g, 10.7 mmol) at 0 °C. After stirring for
1 h at 0 °C, the reaction mixture was diluted
with EtOAc, washed with water and brine,
dried over MgSO4, filtered, concentrated in
vacuo, and chromatographed on a silica-gel
column. Elution with 7:3 cyclohexane–EtOAc
gave 5 (3.02 g, 99%) as an oil. IR wmax(CHCl3)
1
2929, 2857, 1613 cm−1. H NMR (270 MHz,
CDCl3) l 0.88 (3 H, t, J 6.6 Hz), 1.27 (18 H,
bs), 1.43–1.66 (2 H, m), 1.81–1.98 (2 H, m),
2.95 (3 H, s), 3.54 (1 H, m), 3.80 (3 H, s),
4.26–4.51 (4 H, m, containing 2 H, AB-q, J
11.0 Hz, at l 4.38, 4.51), 6.88 (2 H, d, J 8.7
Hz), 7.26 (2 H, d, J 8.6 Hz). FABMS (posi-
1
IR wmax(CHCl3) 2928, 2856, 1616 cm−1. H
NMR (270 MHz, CDCl3) l 0.88 (3 H, t, J 6.5
Hz), 1.19–1.56 (20 H, m), 1.60–1.81 (2 H, m),
3.75–3.82 (m, 4 H, containing 3 H, s, at l

212
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
+
+
’
(28 H, m, containing 3 H, s, at l 1.39, and 3
H, s, at l 1.48), 1.71–1.85 (2 H, m), 2.72 (1 H,
dd, J 3.7, 9.5 Hz), 3.27 (1 H, t, J 9.1 Hz),
3.49–3.86 (9 H, m, containing 3 H, s, at l
3.80), 4.17 (1 H, m), 4.40, 4.47 (2 H, AB-q, J
11.2 Hz), 4.86 (1 H, d, J 3.6 Hz), 5.19–5.34 (2
H, m), 5.88 (1 H, m), 6.87 (2 H, d, J 8.6 Hz),
7.26 (2 H, d, J 8.6 Hz). FABMS (positive-
ion): m/z 592 [M+H]+. HRFABMS (posi-
tive-ion): m/z Calcd for C34H58NO7: 592.4213;
Found: 592.4207. Anal. Calcd for C34H57NO7
(591.8): C, 69.00; H, 9.71; N, 2.37. Found: C,
68.86; H, 10.21; N, 2.31.
tive-ion): m/z 451 [M+Na] , 428 [M ]. HR-
FABMS (positive-ion), Calcd for C23H40O5S:
428.2596; Found: 428.2584.
Allyl
[(R) - 3 - (4 - methoxybenzyloxy)tetradecyl] - 2-
trifluoroacetamide-h- -glucopyranoside (7).—
To a solution of allyl 2-deoxy-4,6-O-isopropy-
lidene-2-trifluoroacetamido-a- -glucopyranosid
2-deoxy-4,6-O-isopropylidene-3-O-
D
D
e (6, 1.28 g, 3.60 mmol) in DMF (15 mL) was
gradually added NaH (55% oil dispersion, 205
mg, 4.70 mmol) at 0 °C with stirring. After 15
min, (R)-3-(4-methoxybenzyloxy)-1-methane-
sulfonyloxytetradecane (1.29 g, 3.00 mmol)
was added to this solution, which was then
stirred for 18 h at rt. The reaction mixture was
quenched with water, extracted with EtOAc,
washed with water and brine, dried over
MgSO4, and filtered. The filtrate was concen-
trated in vacuo to give a mixture that was
chromatographed on a silica-gel column. Elu-
tion with 3:1 cyclohexane–EtOAc gave 7 (1.74
g, 84%) as a wax. IR wmax(CHCl3) 3429, 2929,
Allyl
[(R) - 3 - (4 - methoxybenzyloxy)tetradecyl] - 2-
[(2,2,2 - trichloroethoxycarbonyl)amino] - h -
2-deoxy-4,6-O-isopropylidene-3-O-
D
-
glucopyranoside (9).—To a solution of 8 (1.20
g, 2.03 mmol) in CH2Cl2 (8 mL) were added
satd NaHCO3 (8 mL) and chloroformic acid
2,2,2-trichloroethyl ester (534 mg, 2.52 mmol).
After stirring for 30 min at rt, the mixture was
diluted with EtOAc, washed with water and
brine, dried over MgSO4, filtered, concen-
trated, and chromatographed on a silica-gel
column. Elution with 4:1 cyclohexane–EtOAc
gave 9 (1.47 g, 94%) as a gum. IR wma1x(CHCl3)
3440, 2929, 2857, 1744, 1613 cm−1. H NMR
(270 MHz, CDCl3) l 0.88 (3 H, t, J 6.6
Hz),1.26–1.48 (26 H, m, containing 3 H, s, at
l 1.40, and 3 H, s, at l 1.48), 1.62–1.78 (2 H,
m), 3.38–4.02 (13 H, m, containing 3 H, s, at
l 3.80), 4.16 (1 H, m), 4.39, 4.45 (2 H, AB-q,
J 11.2 Hz), 4.68, 4.74 (2 H, AB-q, J 12.0 Hz),
4.86 (1 H, d, J 3.7 Hz), 5.13–5.32 (3 H, m),
5.89 (1 H, m), 6.88 (2 H, d, J 8.6 Hz), 7.27 (2
H, d, J 8.5 Hz). FABMS (positive-ion): m/z
788 [M+Na]+. FABMS (negative-ion): m/z
764 [M−H]−. HRFABMS (positive-ion): m/z
Calcd for C37H58Cl3NO9: 788.3075; Found:
788.3077. Anal. Calcd for C37H58Cl3NNaO9
(767.2): C, 57.92; H, 7.62; Cl, 13.86; N, 1.83.
Found: C, 57.50; H, 7.38; Cl, 13.87; N, 1.85.
Allyl 2-deoxy-3-O-[(R)-3-(4-methoxybenzy-
loxy)tetradecyl] - 2 - [(2,2,2 - trichloroethoxycar-
2857, 1734 cm−1
.
1H NMR (270 MHz,
CDCl3) l 0.88 (3 H, t, J 6.5 Hz), 1.26–1.49
(26 H, m, containing 3 H, s, at l 1.40, and 3
H, s, at l 1.49), 1.64–1.78 (2 H, m), 3.41–3.91
(11 H, m, containing 3 H, s, at l 3.80), 3.99 (1
H, m), 4.13–4.21 (2 H, m), 4.36, 4.43 (2 H,
AB-q, J 11.2 Hz), 4.87 (1 H, d, J 3.8 Hz),
5.24–5.32 (2 H, m), 5.87 (1 H, m), 6.41 (1 H,
d, J 9.3 Hz, NH), 6.87 (2 H, d, J 8.6 Hz), 7.25
(2 H, d, J 7.9 Hz). FABMS (positive-ion): m/z
710 [M+Na]+. FABMS (negative-ion): m/z
686 [M−H]−. HRFABMS (positive-ion): m/z
Calcd for C36H56F3NNaO8: 710.3856; Found:
710.3851. Anal. Calcd for C36H56F3NO8
(687.8): C, 62.86; H, 8.21; F, 8.19; N, 2.04.
Found: C, 62.56; H, 8.15; F, 7.94; N, 2.07.
Allyl
dene-3-O-[(R)-3-(4-methoxybenzyloxy)tetra-
decyl]-h- -glucopyranoside (8).—A solution
2-amino-2-deoxy-4,6-O-isopropyli-
D
of 7 (1.45 g, 2.11 mmol) in EtOH (10 mL) and
1 M NaOH aq (10 mL) was stirred at 60 °C
for 3 h. The solution was concentrated in
vacuo, diluted with EtOAc, washed with water
and brine, dried over MgSO4, filtered, concen-
trated, and chromatographed on a silica-gel
column. Elution with 2:3 cyclohexane–EtOAc
gave 8 (1.23g, 99%) as a gum. IR wmax(CHCl3)
bonyl)amino]-h-
D
-glucopyranoside
(10).—A
solution of 9 (885 mg, 1.15 mmol) in 80%
AcOH aq (10 mL) was stirred at 60 °C for 1 h.
The solution was diluted with EtOAc, washed
with satd aq NaHCO3 and brine, dried over
MgSO4, filtered, concentrated in vacuo, and
chromatographed on a silica-gel column. Elu-
2928, 2855, 1612 cm−1. H NMR (270 MHz,
1
CDCl3) l 0.88 (3 H, t, J 6.6 Hz), 1.26–1.62

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
213
Allyl 6-O-benzyloxycarbonyl-2-deoxy-4-O-
(diphenylphosphono)-3-O-[(R)-3-(4-methoxy-
benzyloxy)tetradecyl] - 2 - [(2,2,2 - trichloro-
tion with 1:1 cyclohexane–EtOAc gave 10
(696 mg, 83%) as a white powder. IR
wmax(CHCl3) 3601, 3436, 2928, 1742, 1612
1
cm−1. H NMR (270 MHz, CDCl3) l 0.88 (3
ethoxycarbonyl)amino] - h -
D
- glucopyranoside
(12).—To a solution of 11 (390 mg, 0.453
mmol) and DMAP (84 mg, 0.679 mmol) in
CH2Cl2 (3 mL) was added dropwise diphenyl
chlorophosphate (140 mL, 0.675 mmol). After
stirring for 1 h at rt, the reaction mixture was
diluted with EtOAc, washed with satd aq
NaHCO3 and brine, dried over MgSO4,
filtered, and concentrated in vacuo to give a
residue, which was chromatographed on a
silica-gel column. Elution with 3:1 cyclohex-
ane–EtOAc gave 12 (467 mg, 94%) as a gum.
IR wmax(CHCl3) 3435, 2928, 2855, 1747, 1612
H, t, J 6.6 Hz), 1.26 (18 H, bs), 1.43–1.80 (4
H, m), 2.08 (1 H, broad, OH), 3.41–3.54 (4 H,
m), 3.56–3.91 (9 H, m, containing 3 H, s, at l
3.80), 3.99 (1 H, dd, J 6.3, 12.8 Hz), 4.18 (1 H,
dd, J 5.3, 12.8 Hz), 4.39, 4.45 (2 H, AB-q, J
11.2 Hz), 4.68, 4.75 (2 H, AB-q, J 12.0 Hz),
4.85 (1 H, d, J 3.5 Hz), 5.21–5.32 (3 H, m),
5.89 (1 H, m), 6.88 (2 H, d, J 8.8 Hz), 7.26 (2
H, d, J 8.5 Hz). FABMS (positive-ion) 748
[M+Na]+. FABMS (negative-ion): m/z 724
[M−H]−. HRFABMS (positive-ion): m/z
Calcd for C34H54Cl3NNaO9: 748.2762; Found:
748.2744. Anal. Calcd for C34H54Cl3NO9
(727.2): C, 56.16; H, 7.49; Cl, 14.63; N, 1.93.
Found: C, 56.10; H, 7.36; Cl, 14.60; N, 1.96.
Allyl 6-O-benzyloxycarbonyl-2-deoxy-3-O-
[(R) - 3 - (4 - methoxybenzyloxy)tetradecyl] - 2-
1
cm−1. H NMR (270 MHz, CDCl3) l 0.88 (3
H, t, J 6.6 Hz), 1.17–1.72 (22 H, m), 3.37 (1
H, m), 3.65–3.84 (6 H, m, containing 3 H, s,
at l 3.78), 3.93–4.04 (3 H, m), 4.14 (1 H, dd,
J 5.3, 12,9 Hz), 4.20–4.39 (4 H, m), 4.63 (1 H,
m), 4.49, 4.76 (2 H, AB-q, J 12.1 Hz), 4.87 (1
H, d, J 3.6 Hz), 5.06, 5.12 (2 H, AB-q, J 12.2
Hz), 5.21–5.30 (3 H, m), 5.86 (1 H, m), 6.83
(2 H, d, J 8.6 Hz), 7.11–7.37 (17 H, m).
FABMS (positive-ion): m/z 1114 [M+Na]+.
FABMS (negative-ion): m/z 1090 [M−H]−.
HRFABMS (negative-ion): m/z Calcd for
[(2,2,2 - trichloroethoxycarbonyl)amino] - h - -
D
glucopyranoside (11).—To a solution of 10
(366 mg, 0.503 mmol) in CH2Cl2 (5 mL) were
added pyridine (60.4 mg, 0.764 mmol) and
benzyloxycarbonyl chloride (90 mL, 0.630
mmol) at 0 °C under N2. After stirring for 2 h
at rt, the solution was diluted with EtOAc,
washed with 1 N NH4Cl, satd aq NaHCO3,
and brine, dried over MgSO4, and filtered.
The filtrate was concentrated in vacuo, and
chromatographed on a silica-gel column. Elu-
tion with 3:1 cyclohexane–EtOAc gave 11
(419 mg, 97%) as a wax. IR wmax(CHCl3) 3436,
C54H68Cl3NO14P:
1090.3443;
Found:
1090.3431. Anal. Calcd for C54H69Cl3NO14P
(1093.5): C, 59.32; H, 6.36; Cl, 9.73; N, 1.28;
P, 2.83. Found: C, 59.35; H, 6.35; Cl, 9.80; N,
1.21; P, 3.04.
Allyl 6-O-benzyloxycarbonyl-2-deoxy-4-O-
(diphenylphosphono) - 3 - O - [(R) - 3 - (hydroxy)-
tetradecyl]-2-[(2,2,2-trichloroethoxycarbonyl)-
1
2928, 2856, 1744, 1612 cm−1. H NMR (270
MHz, CDCl3) l 0.88 (3 H, t, J 6.6 Hz),
1.26–1.78 (22 H, m), 3.37–3.53 (4 H, m,
containing OH), 3.69–3.99 (8 H, m, contain-
ing 3 H, s, at l 3.79 ppm), 4.15 (1 H, dd, J
5.3, 13.0 Hz), 4.37–4.47 (4 H, m), 4.67, 4.74 (2
H, AB-q, J 12.1 Hz), 4.84 (1 H, d, J 3.7 Hz),
5.18–5.30 (5 H, m), 5.87 (1 H, m), 6.87 (2 H,
d, J 8.6 Hz), 7.25 (2 H, d, J 7.3 Hz), 7.33–7.41
(5 H, m). FABMS (positive-ion): m/z 882
[M+Na]+. FABMS (negative-ion): m/z 858
[M−H]−. HRFABMS (negative-ion): m/z
Calcd for C42H59Cl3NO11: 858.3154; Found:
858.3133. Anal. Calcd for C42H60Cl3NO11
(861.3): C, 58.57; H, 7.02; Cl, 12.35; N, 1.63.
Found: C, 58.38; H, 6.93; Cl, 12.58; N, 1.61.
amino]-h- -glucopyranoside (13).—A mixture
D
of 12 (338 mg, 0.309 mmol), water (0.3 mL),
and DDQ (85 mg, 0.373 mmol) in CH2Cl2 (3
mL) was stirred for 1 h at rt. The mixture was
diluted with EtOAc, washed with satd aq
NaHCO3 and brine, dried over MgSO4,
filtered, concentrated in vacuo, and chro-
matographed on a silica-gel column. Elution
with 7:3 cyclohexane–EtOAc gave 13 (283
mg, 94%) as a gum. IR wmax(CHCl3) 3435,
1
2927, 2855, 1746, 1592 cm−1. H NMR (270
MHz, CDCl3) l 0.88 (3 H, t, J 6.6 Hz),
1.25–1.75 (22 H, m), 3.64–3.85 (4 H, m),
3.95–4.08 (3 H, m), 4.15 (1 H, dd, J 5.3, 12.7
Hz), 4.23–4.38 (2 H, m), 4.63–4.75 (3 H, m),

214
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
complex. Immediately after 2 min, when the
red color solution of iridium complex had
become almost colorless, the hydrogen was
completely replaced with N2. This solution
was stirred for 3 h at rt. After confirming a
double bond shift to an enol ether from the
1-allyloxy group (as indicated by a slightly
higher Rf value on the TLC), water (0.3 mL),
pyridine (15 mg), and I2 (22 mg) were added
to this solution. After 1 h stirring at rt, the
mixture was diluted with EtOAc, washed with
aq 10% Na2S2O3, satd NaHCO3, and brine,
dried over MgSO4, and concentrated to give a
mixture that was chromatographed on a silica-
gel column. Elution with 7:3 cyclohexane–
EtOAc gave 15 (106 mg, 81%) as a gum. IR
wmax(CHCl3) 3599, 3434, 3326, 2927, 2855,
4.93 (1 H, d, J 3.6 Hz), 5.04, 5.11 (2 H, AB-q,
J 12.2 Hz), 5.22–5.31 (2 H, m), 5.44 (1 H, d,
J 9.2 Hz, NH), 5.88 (1 H, m), 7.11–7.34 (15
H, m). FABMS (positive-ion): m/z 994 [M+
Na]+; 972 [M+H]+. HRFABMS (positive-
ion): m/z Calcd for C46H62Cl3NO13P:
972.3024; Found: 972.3010. Anal. Calcd for
C46H61Cl3NO13P (973.3): C, 56.77; H, 6.32; Cl,
10.93; N, 1.44; P, 3.18. Found: C, 56.60; H,
6.49; Cl, 11.17; N, 1.45; P, 3.33.
Allyl 6-O-benzyloxycarbonyl-2-deoxy-4-O-
(diphenylphosphono)-3-O-[(R)-3-(tetradecan-
oyloxy)tetradecyl] - 2 - [(2,2,2 - trichloroethoxy-
carbonyl)amino]-h- -glucopyranoside (14).—
D
To a solution of 13 (271 mg, 0.278 mmol) in
THF (3 mL) were added Et3N (60 mL, 0.427
mmol), DMAP (51 mg, 0.420 mmol), and
tetradecanoyl chloride (0.12 mL, 0.438 mmol)
with stirring. After stirring for 18 h at rt, the
mixture was diluted with EtOAc, washed with
satd aq NaHCO3 and brine, dried over
MgSO4, filtered, concentrated, and chro-
matographed on a silica-gel column. Elution
with 4:1 cyclohexane–EtOAc gave 14 (305
mg, 93%) as a gum. IR wmax(CHCl3) 3435,
1
1746 cm−1. H NMR (270 MHz, CDCl3) l
0.88 (3 H, t, J 6.6 Hz), 1.20–1.77 (44 H, m),
2.21 (2 H, t, J 7.5 Hz), 3.41–3.53 (2 H, m,
containing OH), 3.75 (1 H, m), 3.83–3.95 (2
H, m), 4.19–4.43 (3 H, m), 4.58–4.79 (3 H,
m), 4.92 (1 H, m), 5.04, 5.11 (2 H, AB-q, J
12.1 Hz), 5.29 (1 H, m), 6.03 (1 H, d, J 8.6 Hz,
NH), 7.12–7.34 (15 H, m). FABMS (positive-
ion): m/z 1142 [M+H]+. HRFABMS (posi-
tive-ion): m/z Calcd for C57H83Cl3NNaO14P:
1164.4518; Found: 1164.4518. Anal. Calcd for
C57H83Cl3NO14P (1143.6): C, 59.87; H, 7.37;
Cl, 9.03; N, 1.23; P, 2.71. Found: C, 59.84; H,
7.29; Cl, 9.05; N, 1.46; P, 2.76.
1
2927, 2855, 1746 cm−1. H NMR (270 MHz,
CDCl3) l 0.88 (3 H, t, J 6.6 Hz), 1.19–1.71
(44 H, m), 2.20 (2 H, t, J 7.5 Hz), 3.50 (1 H,
m), 3.68 (1 H, m), 3.82 (1 H, m), 3.92–4.01 (3
H, m), 4.15 (1 H, dd, J 5.3, 13.0 Hz), 4.28–
4.41 (2 H, m), 4.63 (1 H, m), 4.75 (2 H, s),
4.84–4.93 (2 H, m, containing 1 H, d, J 3.6
Hz, at l 4.90), 5.05, 5.11 (2 H, AB-q, J 12.2
Hz), 5.20–5.34 (2 H, m), 5.78 (1 H, d, J 10.7
Hz, NH), 5.87 (1 H, m), 7.12–7.34 (15 H, m).
FABMS (positive-ion): m/z 1182 [M+H]+.
HRFABMS (positive-ion): m/z Calcd for
Allyl 6-O-(tert-butyldimethylsilyl)-2-deoxy-
3-O-[(R)-3-(4-methoxybenzyloxy)tetradecyl]-
2-[(2,2,2-trichloroethoxycarbonyl)amino]-h- -
D
glucopyranoside (16).—To a solution of 15
(1.15 g, 1.58 mmol) in CH2Cl2 (5 mL) were
added DMAP (304 mg, 2.49 mmol) and tert-
butyldimethylsilyl chloride (364 mg, 2.42
mmol). After stirring for 3 h at rt, the mixture
was diluted with EtOAc, washed with water,
satd aq NaHCO3, and brine, dried over
MgSO4, and filtered. The filtrate was concen-
trated in vacuo, and chromatographed on a
silica-gel column. Elution with 4:1 cyclohex-
ane–EtOAc gave 16 (1.28 g, 96%) as a gum.
IR wmax(CHCl3) 3437, 2955, 2929, 2856, 1742,
C60H88Cl3NO14P:
1182.5008;
Found:
1182.5021. Anal. Calcd for C60H87Cl3NO14P
(1183.7): C, 60.88; H, 7.41; Cl, 8.99; N, 1.18;
P, 2.62. Found: C, 61.12; H, 7.46; Cl, 9.18; N,
1.13; P, 2.72.
6-O-Benzyloxycarbonyl-2-deoxy-4-O-(di-
phenylphosphono)-3-O-[(R)-3-(tetradecanoyl-
oxy)tetradecyl] - 2 - [(2,2,2 - trichloroethoxycar-
bonyl)amino]- -glucopyranose (15).—To a so-
D
1
lution of 14 (136 mg, 0.114 mmol) in THF (3
mL) was added [C8H12Ir(PMePh2)2]PF6 (3.6
mg). The air in the reaction flask was com-
pletely replaced with N2 and then further re-
placed with hydrogen to activate the iridium
1613 cm−1. H NMR (270 MHz, CDCl3) l
0.09 (6 H, s), 0.86–0.91 (12 H, m), 1.26 (18 H,
bs), 1.40–1.80 (4 H, m), 3.34–3.67 (4 H, m,
containing OH), 3.76–3.93 (9 H, m, contain-
ing 3 H, s, at l 3.80), 3.98 (1 H, dd, J 6.3, 12.8

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
215
mixture was diluted with EtOAc, washed with
satd aq NaHCO3 and brine, dried over
MgSO4, filtered, concentrated in vacuo, and
chromatographed on a silica-gel column. Elu-
tion with 3:1 cyclohexane–EtOAc gave 18
(1.22 g, 91%) as a gum. IR wmax(CHCl3) 3434,
Hz), 4.18 (1 H, dd, J 5.2, 12.9 Hz), 4.40, 4.45
(2 H, AB-q, J 11.5 Hz), 4.70 (2 H, s), 4.84 (1
H, d, J 3.6 Hz), 5.19–5.32 (3 H, m, containing
NH), 5.90 (1 H, m), 6.87 (2 H, d, J 8.6 Hz),
7.26 (2 H, d, J 8.6 Hz). FABMS (positive-
ion): m/z 878 [M+K]+. FABMS (negative-
ion): m/z 838 [M−H]−. HRFABMS
(negative-ion): m/z Calcd for C40H68Cl3-
NNaO9Si: 862.3627; Found: 862.3622. Anal.
Calcd for C40H68Cl3NO9Si (841.4): C, 57.10;
H, 8.15; Cl, 12.64; N, 1.67. Found: C, 57.54;
H, 8.13; Cl, 12.72; N, 1.57.
1
2928, 2856, 1740 cm−1. H NMR (270 MHz,
CDCl3) l 0.002 (6 H, s), 0.82–0.90 (12 H, m),
1.25–1.67 (22 H, m), 3.65–3.87 (8 H, m,
containing OH), 3,93–4.04 (2 H, m), 4.21 (1
H, dd, J 5.3, 12.9 Hz), 4.62 (1 H, m), 4.75 (2
H, s), 4.93 (1 H, d, J 3.7 Hz), 5.22–5.34 (2 H,
m), 5.44 (1 H, d, J 9.3 Hz, NH), 5.91 (1 H,
m), 7.15–7.35 (10 H, m). FABMS (positive-
ion): m/z 974 [M+Na]+; 952 [M+H]+. HR-
FABMS (positive-ion): m/z Calcd for
Allyl 6-O-tert-butyldimethylsilyl-2-deoxy-4-
O - (diphenylphosphono) - 3 - O - [(R) - 3 - (4-
methoxybenzyloxy)tetradecyl] - 2 - [(2,2,2 - tri-
chloroethoxycarbonyl)amino]-h- -glucopyran-
D
oside (17).—To a solution of 16 (1.26 g, 1.50
mmol) in CH2Cl2 (5 mL) were added DMAP
(278 mg, 2.28 mmol) and diphenyl
chlorophosphate (0.46 mL, 2.23 mmol). After
stirring for 1 h at rt, the mixture was diluted
with EtOAc, washed with satd aq NaHCO3
and brine, dried over MgSO4, and filtered.
The filtrate was concentrated in vacuo, and
chromatographed on a silica-gel column. Elu-
tion with 4:1 cyclohexane–EtOAc gave 17
(1.57 g, 98%) as an oil. IR wmax(CHCl3) 3435,
C44H69Cl3NNaO11PSi:
974.3345.
Allyl 6-O-(tert-butyldimethylsilyl)-2-deoxy-
4-O-(diphenylphosphono)-3-O-[(R)-3-(tetra-
decanoyloxy)tetradecyl] - 2 - [(2,2,2 - trichloro-
974.3341;
Found:
ethoxycarbonyl)amino] - h -
D
- glucopyranoside
(19).—To a solution of 18 (1.21 g, 1.27 mmol)
in THF (6 mL) were added Et3N (2.55 mL,
2.56 mmol), DMAP (326 mg, 2.67 mmol), and
tetradecanoyl chloride (0.70 mL, 2.55 mmol)
with stirring. After stirring for 24 h at rt, the
mixture was diluted with EtOAc, washed with
water, satd aq NaHCO3, and brine, dried over
MgSO4, filtered, concentrated, and chro-
matographed on a silica-gel column. Elution
with 4:1 cyclohexane–EtOAc gave 19 (1.40 g,
95%) as an oil. IR wmax(CHCl3) 3435, 2928,
1
2955, 2929, 2856, 1743, 1613 cm−1. H NMR
(270 MHz, CDCl3) l 0.002 (6 H, s), 0.86–0.92
(12 H, m), 1.26 (18 H, bs), 1.66–1.74 (4 H,
m), 3.37 (1 H, m), 3.69–3.87 (9 H, m, contain-
ing 3 H, s, at l 3.79), 3.94–4.04 (2 H, m), 4.22
(1 H, dd, J 5.3, 12.7 Hz), 4.30 (2 H, s),
4.43–4.61 (2 H, m), 4.75 (1 H, m) 4.89 (1 H,
d, J 3.6 Hz), 5.20–5.34 (3 H, m, containing
NH), 5.91 (1 H, m), 6.84 (2 H, d, J 8.6 Hz),
7.14–7.33 (12 H, m). FABMS (positive-ion):
m/z 1094 [M+Na]+; 1072 [M+H]+. HR-
FABMS (positive-ion): m/z Calcd for
C52H77Cl3NNaO12PSi: 1094.3916; Found:
1094.3905. Anal. Calcd for C52H77Cl3NO12PSi
(1073.6): C, 58.18; H, 7.23; Cl, 9.91; N, 1.31;
P, 2.89. Found: C, 57.85; H, 7.05; Cl, 10.57;
N, 1.27; P, 2.16.
2856, 1737 cm−1
.
1H NMR (270 MHz,
CDCl3) l 0.01 (6 H, s), 0.86–0.92 (15 H, m),
1.26 (40 H, bs), 1.53–1.74 (4 H, m), 2.18 (2 H,
t, J 7.3–7.7 Hz), 3.55 (1 H, m), 3.67–4.07 (7
H, m), 4.22 (1 H, dd, J 5.2–12.8 Hz), 4.56 (1
H, m), 4.77 (2 H, s), 4.83–4.91 (2 H, m,
containing 1 H, d, J 3.6 Hz, at l 4.91),
5.22–5.36 (2 H, m), 5.68 (1 H, d, J 9.4 Hz,
NH), 5.92 (1 H, m), 7.15–7.35 (10 H, m).
FABMS (positive-ion): m/z 1162 [M+H]+;
1200 [M+K]+ (on addition of KI). HR-
FABMS (positive-ion): m/z Calcd for
Allyl 6-O-(tert-butyldimethylsilyl)-2-deoxy-
4 - O - (diphenylphosphono) - 3 - O - [(R)-3-
(hydroxy)tetradecyl]-2-[(2,2,2-trichloroethoxy-
C58H95Cl3KNO12PSi:
1200.5026.
Allyl 2-deoxy-4-O-(diphenylphosphono)-3-
O - [(R) - 3 - (tetradecanoyloxy)tetradecyl] - 2-
1200.5064;
Found:
carbonyl)amino]-h- -glucopyranoside (18).—A
D
mixture of 17 (1.50 g, 1.40 mmol), water (0.5
mL), and DDQ (386 mg, 1.70 mmol) in
CH2Cl2 (5 mL) was stirred for 1 h at rt. The
[(2,2,2 - trichloroethoxycarbonyl)amino] - h - -
D

216
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
1062 [M+H]+. HRFABMS (positive-ion): m/
z Calcd for C53H83Cl3NNaO12P: 1084.4616;
glucopyranoside (20).—To a solution of 19
(1.35 g, 1.16 mmol) in THF (9 mL) was added
aq 3 M HCl (1.2 mL) with stirring. After 2 h
at rt, the reaction mixture was diluted with
EtOAc, washed with satd aq NaHCO3 and
brine, dried over MgSO4, filtered, concen-
trated, and chromatographed on a silica-gel
column. Elution with 7:3 cyclohexane–EtOAc
gave 20 (1.11 g, 91%) as a gum. IR
wmax(CHCl3) 3691, 3606, 3437, 2927, 2855,
Found:
1084.4600.
Anal.
Calcd
for
C53H83Cl3NO12P (1063.6): C, 59.85; H, 7.87;
Cl, 10.00; N, 1.32; P, 2.91. Found: C, 60.14;
H, 7.59; Cl, 10.08; N, 1.42; P, 2.92.
2 - Deoxy - 4 - O - (diphenylphosphono) - 6 - O-
methyl - 3 - O - [(R) - 3 - (tetradecanoyloxy)tetra-
decyl]-2-[(2,2,2-trichloroethoxycarbonyl)amino]-
h- -glucopyranose (22).—Compound 21 (478
D
1
1737 cm−1. H NMR (400 MHz, CDCl3) l
mg, 0.450 mmol) was treated as described in
the formation of 15 from 14 to give 22 (334
mg, 73%) as a solid. IR wmax(KBr) 3436, 3352,
0.88 (6 H, t, J 6.6 Hz), 1.19–1.73 (44 H, m),
2.23 (2 H, t, J 7.3–8.1 Hz), 3.52 (1 H, m),
3.60–3.70 (4 H, m), 3.89–4.03 (3 H, m), 4.17
(1 H, dd, J 5.1, 12.5 Hz), 4.65 (1 H, m), 4.73,
4.78 (2 H, AB-q, J 12.5 Hz), 4.91–4.95 (2 H,
m, containing 1 H, d, J 3.7 Hz, at l 4.95),
5.23–5.35 (2 H, m), 5.82 (1 H, d, J 9.5 Hz,
NH), 5.89 (1 H, m), 7.20–7.38 (10 H, m).
FABMS (positive-ion): m/z 1070 [M+Na]+.
HRFABMS (positive-ion): m/z Calcd for
1
2953, 2921, 2851, 1726 cm−1. H NMR (400
MHz, CDCl3) l 0.88 (6 H, t, J 6.6–7.3 Hz),
1.19–1.73 (44 H, m), 2.19 (2 H, t, J 7.3 Hz),
3.22 (3 H, s), 3.45–3.56 (3 H, m), 3.72–3.94 (4
H, m, containing OH), 4.13 (1 H, m), 4.56 (1
H, m), 4.71, 4.77 (2 H, AB-q, J 12.5 Hz), 4.90
(1 H, m), 5.29 (1 H, m), 5.88 (1 H, d, J 8.8 Hz,
NH), 7.17–7.35 (10 H, m). FABMS (positive-
ion): m/z 1060 [M+K]+ (on addition of KI);
1044 [M+Na]+; 1022 [M+H]+. HRFABMS
(positive-ion): m/z Calcd for C50H79Cl3-
KNO12P: 1060.4043; Found: 1060.4039. Anal.
Calcd for C50H79Cl3NO12P (1023.5): C, 58.68;
H, 7.78; Cl, 10.39; N, 1.37; P, 3.03. Found: C,
58.37; H, 7.62; Cl, 10.47; N, 1.35; P, 2.95.
Allyl 2,6-dideoxy-4-O-((diphenylphosphono))-
6-fluoro-3-O-[(R)-3-(tetradecanoyloxy)tetra-
decyl]-2-[(2,2,2-trichloroethoxycarbonyl)amino]-
C52H81Cl3NNaO12P:
1070.4460;
Found:
1070.4452. Anal. Calcd for C52H81Cl3NO12P
(1049.5): C, 59.51; H, 7.78; Cl, 10.13; N, 1.34;
P, 2.95. Found: C, 59.30; H, 7.90; Cl, 10.24;
N, 1.51; P, 2.94.
Allyl 2-deoxy-4-O-(diphenylphosphono)-6-
O - methyl - 3 - O - [(R) - 3 - (tetradecanoyloxy)-
tetradecyl]-2-[(2,2,2-trichloroethoxycarbonyl)-
amino]-h- -glucopyranoside (21).—To a solu-
D
tion of 20 (1.05 g, 1.00 mmol) in CH2Cl2 (5
mL) were added 2,6-di-tert-butyl-4-methyl-
pyridine (679 mg, 3.31 mmol) and trimethy-
loxonium tetrafluoroborate (446 mg, 3.02
mmol) at 0 °C. After stirring for 3 h at rt, the
reaction mixture was diluted with CH2Cl2,
washed with satd aq NaHCO3 and brine,
dried over MgSO4, filtered, concentrated, and
chromatographed on a silica-gel column. Elu-
tion with 7:3 cyclohexane–EtOAc gave 21
(966 mg, 91%) as a gum. IR wmax(CHCl3) 3435,
h- -glucopyranoside (23).—To a solution of
D
20 (938 mg, 0.894 mmol) in DME (5 mL) was
gradually added a solution of DAST (0.35
mL, 2.65 mmol) in DME (1 mL) at 0 °C with
stirring under N2. After stirring for 3 h at
0 °C, water was added to the reaction mixture,
and it was extracted with EtOAc. The EtOAc
layer was washed with satd aq NaHCO3 and
brine, dried over MgSO4, and concentrated to
give a crude oil. The crude product was chro-
matographed on a silica-gel column. Elution
with 7:3 cyclohexane–EtOAc gave 23 (753
mg, 80%) as a gum. IR wmax(CHCl3) 3435,
1
2927, 2855, 1738 cm−1. H NMR (400 MHz,
CDCl3) l 0.88 (6 H, t, J 6.6–7.3 Hz), 1.18–
1.72 (44 H, m), 2.18 (2 H, t, J 7.3–8.1 Hz),
3.24 (3 H, s), 3.48–3.59 (3 H, m), 3.69 (1 H,
m), 3.82–3.88 (2 H, m), 3.96–4.04 (2 H, m),
4.20 (1 H, dd, J 5.1, 13.2 Hz), 4.64–4.80 (3 H,
m), 4.88 (1 H, m), 4.94 (1 H, d, J 3.7 Hz),
5.22–5.35 (2 H, m), 5.69 (1 H, d, J 8.8 Hz,
NH), 5.90 (1 H, m), 7.16–7.35 (10 H, m).
FABMS (positive-ion): m/z 1084 [M+Na]+;
1
2927, 2855, 1738, 1601 cm−1. H NMR (400
MHz, CDCl3) l 0.88 (6 H, t, J 6.6–7.3 Hz),
1.19–1.73 (44 H, m), 2.20 (2 H, t, J 7.3 Hz),
3.52 (1 H, m), 3.71 (1 H, t, J 9.5 Hz), 3.81–
4.06 (4 H, m), 4.20 (1 H, m), 4.44–4.65 (3 H,
m), 4.76 (2 H, s), 4.90 (1 H, m), 4.95 (1 H, d,
J 3.7 Hz), 5.23–5.36 (2 H, m), 5.78 (1 H, d, J

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
217
chromatographed on a silica-gel column. Elu-
tion with 7:3 cyclohexane–EtOAc gave 28
(152 mg, 88%) as a gum. IR wmax(CHCl3) 3432,
9.5 Hz, NH), 5.90 (1 H, m), 7.17–7.35 (10 H,
m). FABMS (positive-ion): m/z 1072 [M+
Na]+; 1050 [M+H]+. HRFABMS (positive-
ion): m/z Calcd for C52H80Cl3FNNaO11P:
1072.4416; Found: 1072.4424.
1
3354, 2927, 2855, 1739, 1670 cm−1. H NMR
(400 MHz, CDCl3) l 0.88 (12 H, t, J 6.5–7.1
Hz), 1.00–1.73 (86 H, m), 2.19–2.26 (4 H, m),
3.04 (1 H, bs, OH), 3.15 (1 H, m), 3.38–3.68
(12 H, m), 3.85–3.97 (2 H, m), 4.20 (1 H, dd,
J 5.2, 11.9 Hz), 4.27–4.53 (7 H, m, containing
1 H, d, J 5.5 Hz, at l 4.62), 4.78 (1 H, AB-q,
J 12.1 Hz), 4.94–5.12 (3 H, m, containing 2
H, AB-q, J 12.1 Hz, at l 5.04 and 5.10), 5.97
(1 H, m, NH), 6.80 (1 H, d, J 8.7 Hz, NH),
6.82 (1 H, s), 7.12–7.36 (35 H, m). FABMS
(positive-ion): m/z 2138 [M+Na]+; 2115
[M+H]+. HRFABMS (positive-ion): m/z
Calcd for C119H170Cl3N2NaO22P: 2138.0946;
Found: 2138.0940.
2,6 - Dideoxy - 4 - O - (diphenylphosphono) - 6-
fluoro - 3 - O - [(R) - 3 - (tetradecanoyloxy)tetra-
decyl]-2-[(2,2,2-trichloroethoxycarbonyl)amino]-
h- -glucopyranose (24).—Compound 23 (365
D
mg, 0.347 mmol) was treated as described in
the formation of 15 from 14 to give 24 (256
mg, 73%) as a gum. IR wmax(CHCl3) 3434,
1
3326, 2927, 2855, 1737 cm−1. H NMR (400
MHz, CDCl3) l 0.88 (6 H, t, J 6.6 Hz),
1.20–1.71 (44 H, m), 2.22 (2 H, t, J 7.3–8.1
Hz), 3.38 (1 H, bs, OH), 3.49 (1 H, m),
3.76–3.95 (3 H, m), 4.17 (1 H, m), 4.44–4.65
(3 H, m), 4.72, 4.77 (2 H, AB-q, J 12.1 Hz),
4.92 (1 H, m), 5.36 (1 H, m), 6.07 (1 H, d, J
8.8 Hz, NH), 7.18–7.35 (10 H, m). FABMS
(positive-ion): m/z 1010 [M+H]+. HR-
FABMS (positive-ion): m/z Calcd for C49H77-
Cl3FNO11P: 1010.4284; Found: 1010.4271.
Diphenylmethyl 2,6-anhydro-7-O-[6-O-(ben-
zyloxycarbonyl)-2-deoxy-4-O-(diphenylphos-
phono) - 3 - O - [(R) - 3 - (tetradecanoyloxy)tetra-
decyl]-2-[(2,2,2-trichloroethoxycarbonyl)amino]-
Diphenylmethyl 2,6-anhydro-7-O-[2-deoxy-
4 - O - (diphenylphosphono) - 6 - O - methyl - 3 - O-
[(R)-3-(tetradecanoyloxy)tetradecyl]-2-[(2,2,2-
trichloroethoxycarbonyl)amino] - i -
D
- gluco-
pyranosyl] - 3 - [(R) - 3 - (benzyloxy)tetradecana-
mido] - 4 - O - [(R) - 3 - (benzyloxy)tetradecyl] - 3-
deoxy-
D
-glycero- -ido-heptonate (29).—Com-
D
pound 22 (279 mg, 0.273 mmol) was treated as
described in the formation of 28 from 15 to
give 29 (261 mg, 72%) as a gum after chro-
matography on a silica-gel column eluted by
3:2 hexane–EtOAc. IR wmax(CHCl3) 3431,
i -
tetradecanamido] - 4 - O - [(R) - 3 - (benzyloxy)-
tetradecyl]-3-deoxy- -glycero- -ido-heptonate
D
- glucopyranosyl] - 3 - [(R) - 3 - (benzyloxy)-
D
D
1
(28).—To a solution of 15 (116 mg, 0.102
mmol) in CH2Cl2 (2 mL) were added Cl3CCN
(100 mL, 0.997 mmol) and a catalytic amount
of DBU (5 mg) at 0°C. After stirring for 1 h at
0 °C, the reaction mixture was concentrated in
vacuo, and rapidly chromatographed to re-
move DBU on a short column of silica gel
with 3:1 cyclohexane–EtOAc. The elution was
concentrated in vacuo to give a crude imidate
25 (115 mg), which was immediately used for
the next reaction without further purification.
In a N2 atmosphere, a solution of the thus-ob-
tained imidate 25 (115 mg), diol 1 (80.6 mg,
0.081 mmol), and MS4A in CH2Cl2 (2 mL)
was stirred at rt. After stirring for 1 h, ca.
TMSOTf (5 mL) was added to the mixture at
−78 °C. After stirring for 2 h at −78 °C, the
mixture was quenched with satd aq NaHCO3,
diluted with EtOAc, washed with water and
brine, dried over MgSO4, filtered, and concen-
trated in vacuo to give a mixture, which was
3354, 2927, 2855, 1732, 1669 cm−1. H NMR
(400 MHz, CDCl3) l 0.88 (12 H, t, J 6.6 Hz),
1.25–1.76 (86 H, m), 2.20–2.25 (4 H, m), 3.10
(1 H, s, OH), 3.17–3.25 (4 H, m, containing 3
H, s, at l 3.20), 3.40–3.74 (13 H, m), 3.82 (1
H, m), 3.98 (1 H, m), 4.27–4.37 (3 H, m,
containing 2 H, AB-q, J 11.7 Hz, at l 4.29
and 4.33), 4.40, 4.44 (2 H, AB-q, J 11.7 Hz),
4.51 (1 H, m), 4.62–4.65 (3 H, m, containing
1 H, d, J 5.1 Hz, at l 4.62), 4.78 (1 H, AB-q,
J 11.7 Hz), 4.91 (1 H, m), 5.01 (1 H, m), 5.88
(1 H, m, NH), 6.78 (1 H, d, J 8.1 Hz, NH),
6.82 (1 H, s), 7.17–7.35 (30 H, m). FABMS
(positive-ion): m/z 2017 [M+Na]+; 1995
[M+H]+. HRFABMS (positive-ion): m/z
Calcd for C112H166Cl3N2NaO20P: 2018.0735;
Found: 2018.0732.
Diphenylmethyl
2,6 - anhydro - 7 - O - [2,6-
dideoxy-4-O-(diphenylphosphono)-6-fluoro- 3-
O - [(R) - 3 - (tetradecanoyloxy)tetradecyl] - 2-
[(2,2,2 - trichloroethoxycarbonyl)amino] - i - -
D

218
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
(7 H, m), 4,63 (1 H, d, J 5.7 Hz), 5.02–5.10 (3
H, m, containing 2 H, AB-q, J 12.1 Hz, at l
5.03 and 5.10), 5.27 (1 H, d, J 8.2 Hz), 6.64 (1
H, d, J 6.3 Hz, NH), 6.78 (1 H, d, J 8.9 Hz,
NH), 6.81 (1 H, s), 7.12–7.37 (35 H, m).
FABMS (positive-ion): m/z 2006 [M+Na]+;
1984 [M+H]+. HRFABMS (positive-ion):
m/z Calcd for C118H171N2NaO21P: 2006.2010;
Found: 2006.2009.
glucopyranosyl] - 3 - [(R) - 3 - (benzyloxy)tetra-
decanamido] - 4 - O - [(R) - 3 - (benzyloxy)tetra-
decyl] - 3 - deoxy -
D
- glycero -
D
- ido - heptonate
(30).—Compound 24 (255 mg, 0.252 mmol)
was treated as described in the formation of
28 from 15 to give 30 (261 mg, 72%) as a gum
after chromatography on a silica-gel column
eluted by 3:2 hexane–EtOAc. IR wma1x(CHCl3)
2927, 2855, 1732, 1670, 1601 cm−1. H NMR
(400 MHz, CDCl3) l 0.88 (12 H, t, J 6.6 Hz),
1.25–1.71 (86 H, m), 2.21–2.26 (4 H, m), 2.97
(1 H, s, OH), 3.17 (1 H, m), 3.43–4.01 (14 H,
m), 4.27–4.81 (11 H, m, containing 1 H, d, J
5.9 Hz, at l 4.63), 5.01 (1 H, m), 5.94 (1 H, bs,
NH), 6.81 (1 H, d, J 11.0 Hz, NH), 6.82 (1 H,
s), 7.17–7.35 (30 H, m). FABMS (positive-
ion): m/z 2005 [M+Na]+; 1983 [M+H]+.
HRFABMS (positive-ion): m/z Calcd for
C111H163Cl3FN2NaO19P: 2006.0535; Found:
2006.0531.
Diphenylmethyl 2,6-anhydro-7-O-[2-acetam-
ido-2-deoxy-4-O-(diphenylphosphono)-6-O-
methyl - 3 - O - [(R) - 3 - (tetradecanoyloxy)tetra-
decyl]-i-
oxy)tetradecanamido]-4-O-[(R)-3-(benzyloxy)-
tetradecyl]-3-deoxy- -glycero- -ido-heptonate
D
-glucopyranosyl]-3-[(R)-3-(benzyl-
D
D
(32).—Compound 29 (135 mg, 0.068 mmol)
was treated as described in the formation of
31 from 28 to give 32 (81.4 mg, two steps,
65%) as a gum after chromatography on a
silica-gel column eluted by 1:1 hexane–
EtOAc. IR wmax(CHCl3) 3433, 3369, 2927,
Diphenylmethyl 2,6-anhydro-7-O-[2-acetam-
ido-6-O-(benzyloxycarbonyl)-2-deoxy-4-O-
(diphenylphosphono) - 3 - O - [(R) - 3 - (tetrade-
1
2855, 1720, 1669, 1600 cm−1. H NMR (400
MHz, CDCl3) l 0.88 (12 H, t, J 6.6–7.3 Hz),
1.22–1.74 (86 H, m), 1.93 (3 H, s), 2.20–2.24
(4 H, m), 3.18 (1 H, m), 3.22 (3 H, s), 3.37–
3.72 (14 H, m, containing OH), 3.89 (1 H, m),
4.02 (1 H, m), 4.29–4.38 (3 H, m, containing
2 H, AB-q, J 11.0 Hz, at l 4.30 and 4.34), 4.42
(2 H, s), 4.50 (1 H, m), 4.63 (1 H, d, J 5.9 Hz),
5.07 (1 H, m), 5.21 (1 H, d, J 8.1 Hz), 6.56 (1
H, d, J 6.6 Hz, NH), 6.76 (1 H, d, J 8.8 Hz,
NH), 6.81 (1 H, s), 7.17–7.36 (30 H, m).
FABMS (positive-ion): m/z 1885 [M+Na]+.
HRFABMS (positive-ion): m/z Calcd for
canoyloxy)tetradecyl]-i-
[(R) - 3 - (benzyloxy)tetradecanamido] - 4 - O-
[(R) - 3 - (benzyloxy)tetradecyl] - 3 - deoxy -
glycero- -ido-heptonate (31).—To a solution
D
-glucopyranosyl]-3-
D
-
D
of 28 (60.8 mg, 0.029 mmol) in AcOH (2 mL)
was added zinc dust (40.2 mg). After stirring
vigorously for 24 h at rt, the solution was
filtered through Celite to remove the zinc dust,
and concentrated in vacuo to give a crude
product. The product was diluted with
EtOAc, washed with satd aq NaHCO3 and
brine, dried over Na2SO4, filtered, and concen-
trated in vacuo. Next, the crude product was
dissolved in THF (0.4 mL) and water (0.8
mL), pyridine (10 mL) and Ac2O (10 mL)
were added to this solution. After stirring for
4 h at rt, the mixture was diluted with EtOAc,
washed with water and brine, dried over
MgSO4, filtered, concentrated in vacuo, and
chromatographed on a silica-gel column. Elu-
tion with 3:2 cyclohexane–EtOAc gave 31
(40.2 mg, two steps, 70%) as a gum. IR
wmax(CHCl3) 3432, 3365, 2927, 2855, 1738,
C111H167N2NaO19P:
1886.1825.
1886.1798;
Found:
Diphenylmethyl 2,6-anhydro-7-O-[2-acetam-
ido-2,6-dideoxy- 4-O-(diphenylphosphono)-6-
fluoro - 3 - O - [(R) - 3 - (tetradecanoyloxy)tetra-
decyl]-i-
oxy)tetradecanamido]-4-O-[(R)-3-(benzyloxy)-
tetradecyl]-3-deoxy- -glycero- -ido-heptonate
D
-glucopyranosyl]-3-[(R)-3-(benzyl-
D
D
(33).—Compound 30 (152 mg, 0.077 mmol)
was treated as described in the formation of
31 from 28 to give 33 (94.2 mg, two steps,
66%) as a gum after chromatography on a
silica-gel column eluted by 3:2 hexane–
EtOAc. IR wmax(CHCl3) 3432, 3364, 2927,
1
1668 cm−1. H NMR (400 MHz, CDCl3) l
0.88 (12 H, t, J 6.6–7.3 Hz), 1.00–1.76 (86 H,
m), 1.92 (3 H, s), 2.17–2.26 (4 H, m), 3.08 (1
H, m), 3.33–3.72 (12 H, m), 3.89–4.02 (2 H,
m), 4.21 (1 H, dd, J 5.1, 11.9 Hz), 4.28–4.52
1
2855, 1718, 1668, 1600 cm−1. H NMR (400
MHz, CDCl3) l 0.88 (12 H, t, J 6.6 Hz),
1.22–1.73 (86 H, m), 2.21–2.24 (4 H, m), 3.10

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
219
(1 H, m), 3.39–3.69 (12 H, m, containing
OH), 3.93–4.05 (2 H, m), 4.28–4.58 (8 H, m,
containing 2 H, AB-q, J 11.4 Hz, at l 4.30,
4.34, and 2 H, s, at l 4.43), 4.63 (1 H, d, J 5.9
Hz), 5.07 (1 H, m), 5.28 (1 H, d, J 8.8 Hz),
6.59 (1 H, d, J 6.6 Hz, NH), 6.78 (1 H, d, J
8.8 Hz, NH), 6.81 (1 H, s), 7.17–7.35 (30 H,
m). FABMS (positive-ion): m/z 1873 [M+
Na]+; 1851 [M+H]+. HRFABMS (positive-
ion): m/z Calcd for C110H164FN2NaO18P:
1874.1599; Found: 1874.1609.
MHz, CD3OD) l 0.90 (12 H, t, J 6.6–7.3 Hz),
1.28–1.76 (86 H, m), 2.00 (3 H, s), 2.19 (2 H,
t, J 7.3 Hz), 2.34–2.37 (2 H, m), 3.21 (3 H, s),
3.43–3.48 (3 H, m), 3.57–4.01 (13 H, m), 4.21
(1 H, m), 4.47 (1 H, d, J 5.1 Hz), 4.55 (1 H,
m), 4.64 (1 H, d, J 8.1 Hz), 4.83 (1 H, m),
7.20–7.41 (10 H, m). FABMS (positive-ion):
m/z 1539 [M+Na]+. HRFABMS (positive-
ion): m/z Calcd for C84H145N2NaO19P:
1540.0095; Found: 1540.0077.
2,6-Anhydro-7-O-[2-acetamido-2,6-dideoxy-
4-O-(diphenylphosphono)-6-fluoro-3-O-[(R)-
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-4-O-
(diphenylphosphono)-3-O-[(R)-3-(tetradecan-
3 - (tetradecanoyloxy)tetradecyl] - i -
D
- gluco-
oyloxy)tetradecyl] - i -
deoxy - 3 - [(R) - 3 - (hydroxy)tetradecanamido]-
4-O-[(R)-3-(hydroxy)tetradecyl]- -glycero-
D
- glucopyranosyl] - 3-
pyranosyl]-3-deoxy-3-[(R)-3-(hydroxy)tetra-
decanamido]-4-O-[(R)-3-(hydroxy)tetradecyl]-
D
D
-
D
-glycero- -ido-heptonic acid (36).—Com-
D
ido-heptonic acid (34).—A solution of 31 (38.8
mg, 0.020 mmol) in EtOH (2 mL) containing
20% Pd(OH)2–C (35.2 mg) was stirred vigor-
ously under hydrogen for 14 h at rt. The
reaction mixture was filtered and concentrated
in vacuo to give a crude product. This crude
product was chromatographed on a short
column of silica gel. Elution with 8:1 CHCl3–
MeOH gave 34 (15.2 mg, 52%) as an amor-
phous resin. A fair amount of 34 was
irreversibly absorbed on the silica-gel column.
IR wmax(CHCl3) 3691, 3606, 3415, 3358, 1716,
pound 33 (90.3 mg, 0.049 mmol) was treated
as described in the formation of 34 from 31 to
give 36 (47.1 mg, 64%) as an amorphous solid
after chromatography on a silica-gel column
eluted by 8:1 CHCl3–MeOH. IR wmax(CHCl3)
3606, 3416, 3357, 2927, 2855, 1713, 1663, 1602
cm−1. 1H NMR (400 MHz, CDCl3) l 0.90 (12
H, t, J 6.6 Hz), 1.23–1.75 (86 H, m), 2.00 (3
H, s), 2.20 (2 H, t, J 7.3 Hz), 2.34–2.37 (2 H,
m), 3.45 (1 H, m), 3.58–4.04 (13 H, m), 4.20
(1 H, m), 4.38–4.58 (4 H, m, containing 1 H,
d, J 5.1 Hz, at l 4.47), 4.70 (1 H, d, J 8.1 Hz),
7.19–7.42 (10 H, m). FABMS (positive-ion):
m/z 1527 [M+Na]+. HRFABMS (positive-
ion): m/z Calcd for C83H142FN2NaO18P:
1527.9877; Found: 1527.9941.
1662, 1602 cm−1
.
1H NMR (400 MHz,
CD3OD) l 0.90 (12 H, t, J 6.6 Hz), 1.14–1.76
(86 H, m), 2.01 (3 H, s), 2.19 (2 H, t, J 7.1
Hz), 2.33–2.37 (2 H, m), 3.43 (1 H, m), 3.51–
4.04 (14 H, m), 4.20 (1 H, m), 4.41 (1 H, d, J
4.3 Hz), 4.54 (1 H, m), 4.70 (1 H, m), 4.81–
4.90 (2 H, m), 7.19–7.41 (10 H, m). FABMS
(positive-ion): m/z 1525 [M+Na]+; 1503
[M+H]+. HRFABMS (positive-ion): m/z
Calcd for C83H143N2NaO19P: 1525.9920;
Found: 1525.9941.
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-4-O-
phosphono - 3 - O - [(R) - 3 - (tetradecanoyloxy)-
tetradecyl] - i -
[(R)-3-(hydroxy)tetradecanamido]-4-O-[(R)-
3-(hydroxy)tetradecyl]- -glycero- -ido-hep-
D
- glucopyranosyl] - 3 - deoxy - 3-
D
D
tonic acid (37).—A solution of 34 (3.5 mg,
0.002 mmol) in THF (1 mL) containing PtO2
(3.1 mg) was stirred vigorously under H2 for
18 h at rt. The reaction mixture was filtered
and concentrated in vacuo to give a residue.
The residue was dissolved in CHCl3 (4 mL),
MeOH (8 mL) and aq 0.1 M HCl (3.2 mL).
To this solution was added another volume of
CHCl3 (4 mL) and aq 0.1 M HCl (4 mL) to
separate the solution into two phases. The
lower CHCl3 phase was collected and concen-
trated to give 37 (3.0 mg, 95%) as a white
powder, mp 168–169 °C. [h]2D5 −4.2° (c 0.10,
CHCl3). IR wmax(CHCl3) 3353, 1713, 1657,
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-4-O-
(diphenylphosphono)-6-O-methyl-3-O-[(R)-3-
(tetradecanoyloxy)tetradecyl]-i-
osyl]-3-deoxy-3-[(R)-3-(hydroxy)tetradecana-
mido] - 4 - O - [(R) - 3 - (hydroxy)tetradecyl] -
glycero- -ido-heptonic acid (35).—Compound
D
-glucopyran-
D
-
D
32 (73.1 mg, 0.039 mmol) was treated as de-
scribed in the formation of 34 from 31 to give
35 (42.7 mg, 72%) as an amorphous solid after
chromatography on a silica-gel column eluted
by 8:1 CHCl3–MeOH. IR wmax(KBr) 3321,
1
2924, 2854, 1731, 1651 cm−1. H NMR (400

220
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
1375 [M+Na]+; 1353 [M+H]+. HRFABMS
1604 cm−1
.
1H NMR (400 MHz, 3:1
(positive-ion): m/z Calcd for C71H134FN2-
NaO18P: 1375.9251; Found: 1375.9238. Anal.
Calcd for C71H134FN2O18P (1353.8): C, 62.99;
H, 9.98; F, 1.40; N, 2.07; P, 2.29. Found: C,
62.77; H, 9.72; F, 1.30; N, 2.05; P, 2.28.
Diphenylmethyl 2,6-anhydro-7-O-[6-O-(ben-
zyloxycarbonyl)-2-deoxy-4-O-(diphenylphos-
phono)-2-[(R)-3-(dodecanoyloxy)tetradecana-
mido] - 3 - O - [(R) - 3 - (tetradecanoyloxy)tetra-
CD3OD–CDCl3) l 0.89 (12 H, t, J 6.6–7.3
Hz), 1.28–1.75 (86 H, m), 2.01 (3 H, s),
2.28–2.42 (4 H, m), 3.31–4.53 (20 H, m), 4.99
(1 H, m). FABMS (positive-ion): m/z 1389
[M+K]+ (on addition of KI); 1373 [M+
Na]+; 1351 [M+H]+. HRFABMS (positive-
ion): m/z Calcd for C71H136N2O19P: 1351.9475;
Found: 1351.9468.
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-6-O-
methyl - 4 - O - phosphono - 3 - O - [(R) - 3 - (tetra-
decanoyloxy)tetradecyl]-i-
3-deoxy-3-[(R)-3-(hydroxy)tetradecanamido]-
4-O-[(R)-3-(hydroxy)tetradecyl]- -glycero- -
decyl]-i-
oxy)tetradecanamido]-4-O-[(R)-3-(benzyloxy)-
tetradecyl]-3-deoxy- -glycero- -ido-heptonate
D
-glucopyranosyl]-3-[(R)-3-(benzyl-
D
-glucopyranosyl]-
D
D
(40).—To a solution of 28 (118.2 mg, 0.056
mmol) in AcOH (3 mL) was added zinc dust
(105 mg). After stirring vigorously for 20 h at
rt, the solution was filtered through Celite to
remove the zinc dust and concentrated in
vacuo to give a crude product. The product
was diluted with EtOAc, washed with satd aq
NaHCO3 and brine, dried over Na2SO4,
filtered, and concentrated in vacuo. Next, the
crude product was dissolved in 1,2-dichloro-
ethane, and DCC (25.5 mg, 0.124 mmol) and
(R)-3-(dodecanoyloxy)tetradecanoic acid (48.5
mg, 0.114 mmol) were added. After stirring
for 6 h at rt, the mixture was filtered, concen-
trated in vacuo, and chromatographed on a
silica-gel column. Elution with 3:2 cyclohex-
ane–EtOAc gave 40 (70.6 mg, two steps, 54%)
as a gum. IR wmax(CHCl3) 3692, 3435, 3364,
D
D
ido-heptonic Acid (38).—Compound 35 (40.8
mg, 0.027 mmol) was treated as described in
the formation of 37 from 34 to give 38 (35.4
mg, 96%) as a white powder, mp 177.5–
24
D
178.6 °C. [h]
−44.6° (c 0.12, CHCl3). IR
wmax(KBr) 3315, 2924, 2854, 1730, 1646 cm−1
.
1H NMR (400 MHz, 1:1 CD3OD–CDCl3) l
0.89 (12 H, t, J 6.6–7.3 Hz), 1.23–1.85 (86 H,
m), 2.01 (3 H, s), 2.29–2.42 (4 H, m), 3.41 (3
H, s), 3.48–3.92 (15 H, m), 4.02–4.14 (2 H,
m), 4.28 (1 H, dd, J 5.1, 9.5 Hz), 4.50 (1 H, d,
J 5.1 Hz), 4.62 (1 H, d, J 8.8 Hz), 4.95 (1 H,
m). FABMS (positive-ion): m/z 1387 [M+
Na]+; 1365 [M+H]+. HRFABMS (positive-
ion): m/z Calcd for C72H137N2NaO19P:
1541387.9451; Found: 1387.9464. Anal. Calcd
for C72H137N2O19P (1365.8): C, 63.32; H,
10.11; N, 2.05; P, 2.27. Found: C, 62.95; H,
9.94; N, 2.15; P, 2.27.
1
2928, 2855, 1732, 1660, 1601 cm−1. H NMR
(400 MHz, CDCl3) l 0.88 (18 H, t, J 6.6 Hz),
1.05–1.96 (124 H, m), 2.14 (2 H, t, J 7.3–8.1
Hz), 2.21–2.23 (2 H, m), 2.27 (2 H, t, J
7.3–8.1 Hz), 2.40–2.42 (2 H, m), 3.10 (1 H,
m), 3.39–3.70 (12 H, m, containing OH),
3.90–4.05 (2 H, m), 4.17 (1 H, dd, J 5.1, 11.7
Hz), 4.31 (2 H, s), 4.34–4.37 (2 H, m), 4.41 (2
H, s), 4.48 (1 H, dd, J 8.4, 17.2 Hz), 4.62 (1 H,
d, J 5.1 Hz), 5.01–5.11 (3 H, m, containing 2
H, AB-q, J 11.7 Hz, at l 5.02 and 5.09),
5.22–5.28 (2 H, m, containing 1 H, d, J 8.1
Hz, at l 5.23), 6.69 (1 H, d, J 5.9 Hz), 6.74 (1
H, d, J 9.5 Hz), 6.80 (1 H, s), 7.11–7.34 (35
H, m). FABMS (positive-ion): m/z 2387 [M+
K]+ (on addition of KI); 2371 [M+Na]+.
HRFABMS (positive-ion): m/z Calcd for
2,6-Anhydro-7-O-[2-acetamido-2,6-dideoxy-
6-fluoro-4-O-phosphono-3-O-[(R)-3-(tetra-
decanoyloxy)tetradecyl]-i-
3-deoxy-3-[(R)-3-(hydroxy)tetradecanamido]-
4-O-[(R)-3-(hydroxy)tetradecyl]- -glycero- -
D
-glucopyranosyl]-
D
D
ido-heptonic acid (39).—Compound 36 (45.3
mg, 0.030 mmol) was treated as described in
the formation of 37 from 34 to give 39 (40.1
mg, 98%) as a white powder, mp. 191.5–
192.0 °C. [h]2D4 −30.6° (c 0.10, CHCl3). IR
wmax(KBr) 3304, 2924, 2854, 1730, 1648 cm−1
.
1H NMR (400 MHz, 1:1 CD3OD–CDCl3) l
0.89 (12 H, t, J 6.6 Hz), 1.18–1.83 (86 H, m),
2.01 (3 H, s), 2.30–2.42 (4 H, m), 3.48–3.92
(16 H, m), 4.04–4.14 (2 H, m), 4.27 (1 H, m),
4.50 (1 H, d, J 5.1 Hz), 4.65–4.69 (2 H, m),
4.96 (1 H, m). FABMS (positive-ion): m/z
C142H217KN2O23P:
2388.5266.
2388.5247;
Found:

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
221
H, d, J 6.6 Hz), 6.76 (1 H, d, J 8.8 Hz), 6.81
(1 H, s), 7.15–7.35 (30 H, m). FABMS (posi-
tive-ion): m/z 2239 [M+Na]+, 2217 [M+
H]+. HRFABMS (positive-ion): m/z Calcd
for C134H210FN2NaO20P: 2240.5096; Found:
2240.5071.
Diphenylmethyl 2,6-anhydro-7-O-[2-deoxy-
4-O-(diphenylphosphono)-2-[(R)-3-(dodecan-
oyloxy)tetradecanamido]-6-O-methyl-3-O-[(R)-
3 - (tetradecanoyloxy)tetradecyl] - i -
D
- gluco-
pyranosyl] - 3 - [(R) - 3 - (benzyloxy)tetradecana-
mido] - 4 - O - [(R) - 3 - (benzyloxy)tetradecyl] - 3-
2,6-Anhydro-7-O-[2-deoxy-4-O-(diphenyl-
phosphono)-2-[(R)-3-(dodecanoyloxy)tetrade-
canamido]-3-O-[(R)-3-(tetradecanoyloxy)tetra-
deoxy-
D
-glycero- -ido-heptonate (41).—Com-
D
pound 29 (110.3 mg, 0.055 mmol) was treated
as described in the formation of 40 from 28 to
give 41 (87.6 mg, two steps, 71%) as a gum
after chromatography on a silica-gel column
eluted by 3:2 hexane–EtOAc. IR wmax(CHCl3)
decyl]-i-
(hydroxy)tetradecanamido] - 4 - O - [(R) - 3 -
(hydroxy)tetradecyl]- -glycero- -ido-heptonic
D
-glucopyranosyl]-3-deoxy-3-[(R)-3-
D
D
1
3431, 3364, 2927, 2855, 1727, 1671 cm−1. H
acid (43).—Compound 40 (70.5 mg, 0.030
mmol) was treated as described in the forma-
tion of 34 from 31 to give 43 (27.4 mg, 49%)
as an amorphous solid after chromatography
on a silica-gel column eluted by 9:1 CHCl3–
MeOH. IR wmax(CHCl3) 3418, 2927, 2855,
NMR (400 MHz, CDCl3) l 0.88 (18 H, t, J
6.6 Hz), 1.22–1.75 (124 H, m), 2.16 (2 H, t, J
7.3–8.1 Hz), 2.21–2.23 (2 H, m), 2.27 (2 H, t,
J 7.3 Hz), 2.41–2.43 (2 H, m), 3.16–3.23 (4 H,
m, containing 3 H, s, at l 3.20), 3.41–3.60 (12
H, m, containing OH), 3.69–3.75 (2 H, m),
3.87–3.99 (2 H, m), 4.31 (2 H, s), 4.35 (1 H,
m), 4.42 (2 H, s), 4.47 (1 H, m), 4.62 (1 H, d,
J 5.9 Hz), 5.10 (1 H, m), 5.17 (1 H, d, J 8.1
Hz), 5.27 (1 H, m), 6.66 (1 H, d, J 6.6 Hz),
6.74 (1 H, d, J 8.8 Hz), 6.81 (1 H, s), 7.17–
7.34 (30 H, m). FABMS (positive-ion): m/z
2251 [M+Na]+. HRFABMS (positive-ion):
m/z Calcd for C135H213N2NaO21P: 2252.5296;
Found: 2252.5334.
1
1719, 1665, 1600 cm−1. H NMR (400 MHz,
3:1 CD3OD–CDCl3) l 0.90 (18 H, t, J 6.6–7.3
Hz), 1.28–1.77 (124 H, m), 2.22 (2 H, t, J 7.3
Hz), 2.28–2.41 (4 H, m), 2.49–2.60 (2 H, m),
3.47–4.02 (16 H, m), 4.23 (1 H, m), 4.46 (1 H,
d, J 5.1 Hz), 4.54 (1 H, m), 4.75 (1 H, d, J 8.1
Hz), 4.89 (1 H, m), 5.27 (1 H, m), 7.18–7.40
(10 H, m). FABMS (positive–ion): m/z 1891
[M+Na]+; 1869 [M+H]+. HRFABMS
(positive-ion): m/z Calcd for C107H189N2-
NaO21P: 1892.3418; Found: 1892.3439.
Diphenylmethyl
2,6-anhydro-7-O-[2,6-
2,6-Anhydro-7-O-[2-deoxy-4-O-(diphenyl-
phosphono) - 2 - [(R) - 3 - (dodecanoyloxy)tetra-
decanamido]-6-O-methyl-3-O-[(R)-3-(tetra-
dideoxy-4-O-(diphenylphosphono)-2-[(R)-3-
(dodecanoyloxy)tetradecanamido] - 6 - fluoro - 3-
O-[(R)-3-(tetradecanoyloxy)tetradecyl]-i- -
D
decanoyloxy)tetradecyl]-i-
3-deoxy-3-[(R)-3-(hydroxy)tetradecanamido]-
4-O-[(R)-3-(hydroxy)tetradecyl]- -glycero- -
D
-glucopyranosyl]-
glucopyranosyl]-3-[(R)-3-(benzyloxy)tetrade-
canamido]-4-O-[(R)-3-(benzyloxy)tetradecyl]-
D
D
3-deoxy-
D
-glycero-
D
-ido-heptonate
(42).—
ido-heptonic acid (44).—Compound 41 (80.6
mg, 0.036 mmol) was treated as described in
the formation of 34 from 31 to give 44 (52.5
mg, 77%) as an amorphous solid after chro-
matography on a silica-gel column eluted by
9:1 CHCl3–MeOH. IR wmax(CHCl3) 2927,
Compound 30 (80.1 mg, 0.040 mmol) was
treated as described in the formation of 40
from 28 to give 42 (51.4 mg, two steps, 58%)
as a gum after chromatography on a silica-gel
column eluted by 3:2 hexane–EtOAc. IR
wmax(CHCl3) 3431, 3362, 2927, 2855, 1727,
1
2855, 1716, 1666, 1601 cm−1. H NMR (400
1671, 1601 cm−1
.
1H NMR (400 MHz,
MHz, 3:1 CD3OD–CDCl3) l 0.90 (18 H, t, J
6.6–7.3 Hz), 1.28–1.74 (124 H, m), 2.22 (2 H,
t, J 7.3 Hz), 2.28–2.41 (4 H, m), 2.50–2.59 (2
H, m), 3.20 (3 H, s), 3.44–3.98 (16 H, m), 4.23
(1 H, m), 4.48 (1 H, d, J 5.1 Hz), 4.55 (1 H,
m), 4.73 (1 H, d, J 8.1 Hz), 4.89 (1 H, m), 5.27
(1 H, m), 7.19–7.40 (10 H, m). FABMS (pos-
itive-ion): m/z 1905 [M+Na]+. HRFABMS
(positive-ion): m/z Calcd for C108H191N2-
NaO21P: 1906.3575; Found: 1906.3572.
CDCl3) l 0.88 (18 H, t, J 6.6 Hz), 1.22–1.74
(124 H, m), 2.15 (2 H, t, J 7.3–8.1 Hz),
2.22–2.23 (2 H, m), 2.27 (2 H, t, J 7.3–8.1
Hz), 2.41–2.42 (2 H, m), 3.13 (1 H, m), 3.42–
3.70 (12 H, m, containing OH), 3.93–4.01 (2
H, m), 4.31 (2 H, s), 4.36–4.55 (6 H, m,
containing 2 H, s, at l 4.42), 4.62 (1 H, d, J
5.1 Hz), 5.10 (1 H, m), 5.23–5.28 (2 H, m,
containing 1 H, d, J 8.1 Hz, at l 5.24), 6.69 (1

222
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
3-(hydroxy)tetradecyl]-
D
-glycero- -ido-hep-
D
2,6 - Anhydro - 7 - O - [2,6 - dideoxy - 4 - O - (di-
tonic acid (47).—Compound 44 (80.6 mg,
0.036 mmol) was treated as described in the
formation of 42 from 34 to give 47 (52.5 mg,
77%) as a white powder, mp 176.5–177.0 °C.
[h]2D4 −3.4° (c 0.18, CHCl3). IR wmax(KBr)
phenylphosphono) - 2 - [(R) - 3 - (dodecanoyloxy)-
tetradecanamido]-6-fluoro-3-O-[(R)-3-(tetra-
decanoyloxy)tetradecyl]-i-
3-deoxy-3-[(R)-3-(hydroxy)tetradecanamido]-
4-O-[(R)-3-(hydroxy)tetradecyl]- -glycero- -
D
-glucopyranosyl]-
D
D
3409, 2957, 2922, 2853, 1730, 1649 cm−1. H
1
ido-heptonic acid (45).—Compound 42 (80.1
mg, 0.040 mmol) was treated as described in
the formation of 34 from 31 to give 45 (51.4
mg, two steps, 58%) as an amorphous solid
after chromatography on a silica-gel column
eluted by 9:1 CHCl3–MeOH IR wma1x(CHCl3)
2927, 2855, 1716, 1665, 1602 cm−1. H NMR
(400 MHz, 3:1 CD3OD–CDCl3) l 0.88 (18 H,
t, J 6.6–7.3 Hz), 1.29–1.82 (124 H, m), 2.23 (2
H, t, J 7.3–8.1 Hz), 2.30 (2 H, t, J 7.3–8.1
Hz), 2.34–2.40 (2 H, m), 2.50–2.56 (2 H, m),
3.47–4.01 (14 H, m), 4.22 (1 H, m), 4.34–4.58
(4 H, m, containing 1 H, d, J 5.1 Hz, at l
4.47), 4.76–4.89 (2 H, m, containing 1 H, d, J
8.1 Hz, at l 4.80), 5.28 (1 H, m), 7.18–7.42
(10 H, m). FABMS (positive-ion): m/z 1893
[M+Na]+; 1871 [M+H]+. HRFABMS
(positive-ion): m/z Calcd for C107H188FN2-
NaO20P: 1894.3375; Found: 1894.3374.
NMR (400 MHz, 3:1 CD3OD–CDCl3) l 0.89
(18 H, t, J 6.6 Hz), 1.28–1.85 (124 H, m),
2.29–2.42 (6 H, m), 2.49 (1 H, dd, J 5.9, 14.6
Hz), 2.57 (1 H, dd, J 7.3, 14.6 Hz), 3.40 (3 H,
s), 3.53–3.63 (7 H, m), 3.69 (1 H, m), 3.79–
4.00 (8 H, m), 4.09 (1 H, m), 4.27 (1 H, m),
4.51 (1 H, d, J 5.1 Hz), 4.66 (1 H, d, J 8.1 Hz),
4.98 (1 H, m), 5.26 (1 H, m). FABMS (posi-
tive-ion): m/z 1753 [M+Na]+. FABMS (neg-
ative-ion): m/z 1729 [M−H]−1. HRFABMS
(positive-ion): m/z Calcd for C96H183N2-
NaO21P: 1754.2949; Found: 1754.3015. Anal.
Calcd for C96H183N2O21P (1732.5): C, 66.56;
H, 10.65; N, 1.62; P, 1.79. Found: C, 66.16; H,
10.66; N, 1.54; P, 1.68.
2,6-Anhydro-7-O-[2,6-dideoxy-2-[(R)-3-
(dodecanoyloxy)tetradecanamido] - 6 - fluoro - 4-
O-phosphono-3-O-[(R)-3-(tetradecanoyloxy)-
tetradecyl] - i -
[(R)-3-(hydroxy)tetradecanamido]-4-O-[(R)-
3-(hydroxy)tetradecyl]- -glycero- -ido-hep-
D
- glucopyranosyl] - 3 - deoxy - 3-
2,6-Anhydro-7-O-[2-deoxy-2-[(R)-3-(dode-
canoyloxy)tetradecanamido] - 4 - O - phosphono-
3 - O - [(R) - 3 - (tetradecanoyloxy)tetradecyl] -
D
D
tonic acid (48).—Compound 45 (11.8 mg,
0.006 mmol) was treated as described in the
formation of 37 from 34 to give 48 (9.5 mg,
88%) as a white powder, mp 195.0–196.0 °C.
[h]2D5 −6.2° (c 0.22, CHCl3). IR wmax(KBr)
3423, 3312, 3085, 2957, 2923, 2853, 1731, 1649
i -
(hydroxy)tetradecanamido] - 4 - O - [(R) - 3-
(hydroxy)tetradecyl]- -glycero- -ido-heptonic
D
- glucopyranosyl] - 3 - deoxy - 3 - [(R) - 3-
D
D
acid (46).—Compound 43 (20.1 mg, 0.011
mmol) was treated as described in the forma-
tion of 37 from 34 to give 46 (18.1 mg, 98%)
as a white powder, mp 185.5–186.3 °C. [h]D25
−0.9° (c 0.25, CHCl3). IR wmax(KBr) 3400,
3310, 3085, 2957, 2923, 2853, 1730, 1650
1
cm−1. H NMR (400 MHz, 1:1 CD3OD–
CDCl3) l 0.90 (18 H, t, J 6.6–7.3 Hz), 1.29–
1.87 (124 H, m), 2.30–2.42 (6 H, m), 2.50 (1
H, dd, J 5.9, 14.6 Hz), 2.57 (1 H, dd, J 7.3,
14.6 Hz), 3.52–4.03 (14 H, m), 4.11 (1 H, m),
4.27 (1 H, m), 4.51 (1 H, d, J 5.1 Hz),
4.57–4.73 (3 H, m), 4.99 (1 H, m), 5.26 (1 H,
m). FABMS (positive-ion): m/z 1741 [M+
Na]+. HRFABMS (positive-ion): m/z Calcd
for C95H180FN2NaO20P: 1742.2749; Found:
1742.2716.
1
cm−1. H NMR (400 MHz, 1:1 CD3OD–
CDCl3) l 0.90 (18 H, t, J 6.6–7.0 Hz), 1.12–
1.85 (124 H, m), 2.30–2.57 (8 H, m),
3.34–4.01 (16 H, m), 4.13 (1 H, m), 4.30 (1 H,
m), 4.50 (1 H, d, J 5.0 Hz), 4.68 (1 H, d, J 8.3
Hz), 5.00 (1 H, m), 5.25 (1 H, m). FABMS
(positive-ion): m/z 1739 [M+Na]+. HR-
FABMS (positive-ion): m/z Calcd for C95H181-
N2NaO21P: 1740.2792; Found: 1740.2795.
2,6-Anhydro-7-O-[2-deoxy-2-[(R)-3-(dode-
canoyloxy)tetradecanamido]-6-O-methyl-4-O-
phosphono - 3 - O - [(R) - 3 - (tetradecanoyloxy)-
Allyl
tetradecyl] - 4,6 - O - isopropylidene - 2 - trifluoro-
acetamido-h- -glucopyranoside (49).—To a
solution of allyl 2-deoxy-4,6-O-isopropyli-
2-deoxy-3-O-[(R)-3-(dodecyloxy)-
D
dene-2-trifluoroacetamido-a- -glucopyrano-
D
tetradecyl] - i -
D
- glucopyranosyl] - 3 - deoxy - 3-
side (6, 4.23 g, 11.9 mmol) in DMF (60 mL)
[(R)-3-(hydroxy)tetradecanamido]-4-O-[(R)-
was gradually added NaH (60% oil dispersion,

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
223
4.13 (1 H, dd, J 5.9, 12.5 Hz), 4.74 (2 H, s),
4.87 (1 H, d, J 3.7 Hz), 5.22–5.32 (3 H, m,
containing NH), 5.89 (1 H, m). FABMS (pos-
itive-ion): m/z 814 [M+H]+. HRFABMS
(positive-ion): m/z Calcd for C41H75Cl3NO8:
814.4558; Found: 814.4551. Anal. Calcd for
C41H74Cl3NO8 (815.4): C, 60.39; H, 9.15; Cl,
13.04; N, 1.72. Found: C, 59.94; H, 9.04; Cl,
12.89; N, 1.70.
725 mg, 18.1 mmol) at 0 °C with stirring.
After 30 min, (R)-3-(dodecyloxy)-1-(methane-
sulfonyloxy)tetradecane (6.80 g, 14.3 mmol)
was added to this solution, which was then
stirred at rt for 3 h. The reaction mixture was
quenched with water, extracted with EtOAc,
washed with water and brine, dried over
MgSO4, and filtered. The filtrate was concen-
trated in vacuo to give a mixture that was
chromatographed on a silica-gel column. Elu-
tion with 4:1 hexane–EtOAc gave 49 (6.60 g,
75%) as a wax. IR wmax(CHCl3) 3431, 2928,
Allyl
tetradecyl]-2-[(2,2,2-trichloroethoxycarbonyl)-
amino]-h- -glucopyranoside (51).—A solution
2-deoxy-3-O-[(R)-3-(dodecyloxy)-
D
2855, 1734 cm−1
.
1H NMR (400 MHz,
of 50 (3.51 g, 4.30 mmol) in 80% aq AcOH (20
mL) was stirred at 60 °C for 4 h. The solution
was diluted with EtOAc, washed with satd
aq NaHCO3 and brine, dried over MgSO4,
filtered, concentrated in vacuo, and chro-
matographed on a silica-gel column. Elution
with 3:2 hexane–EtOAc gave 51 (3.15 g, 95%)
as a white powder. IR wmax(KBr) 3335, 2923,
CDCl3) l 0.88 (6 H, t, J 6.6 Hz), 1.21–1.67
(48 H, m, containing 3 H, s, at l 1.41, and 3
H, s, at l 1.51), 3.29–3.41 (3 H, m), 3.49–3.58
(2 H, m), 3.67–3.78 (3 H, m), 3.83–3.89 (2 H,
m), 3.98 (1 H, dd, J 5.1, 12.5 Hz), 4.13–4.20
(2 H, m), 4.89 (1 H, d, J 3.7 Hz), 5.24–5.31 (2
H, m), 5.86 (1 H, m), 6.55 (1 H, d, J 9.5 Hz,
NH). FABMS (positive-ion): m/z 736 [M+
H]+. HRFABMS (positive-ion): m/z Calcd
for C40H73F3NO7: 736.5339; Found: 736.5337.
Anal. Calcd for C40H72F3NO7 (736.0): C,
65.28; H, 9.86; F, 7.74; N, 1.90. Found: C,
65.35; H, 9.89; F, 7.86; N, 1.90.
2853, 1709 cm−1
.
1H NMR (400 MHz,
CDCl3) l 0.88 (6 H, t, J 6.6 Hz), 1.26–1.73
(42 H, m), 2.17 (1 H, t, J 6.6 Hz, OH),
3.36–3.50 (4 H, m), 3.60–3.91 (8 H, m, con-
taining OH), 3.99 (1 H, dd, J 6.6, 13.2 Hz),
4.19 (1 H, dd, J 5.1, 13.2 Hz), 4.69, 4.81 (2 H,
AB-q, J 11.7 Hz), 4.87 (1 H, d, J 3.7 Hz),
5.22–5.32 (3 H, m), 5.90 (1 H, m). FABMS
(positive-ion): m/z 774 [M+H]+. HRFABMS
(positive-ion): m/z Calcd for C38H71Cl3NO8:
774.4245; Found: 774.4228. Anal. Calcd for
C38H70Cl3NO8 (775.3): C, 58.87; H, 9.10; Cl,
13.72 N, 1.81. Found: C, 58.87; H, 8.94; Cl,
14.00; N, 1.81.
Allyl
tetradecyl]-4,6-O-isopropylidene-2-[(2,2,2-tri-
chloroethoxycarbonyl)amino]-h- -glucopyran-
2-deoxy-3-O-[(R)-3-(dodecyloxy)-
D
oside (50).—A solution of 41 (4.70 g, 6.39
mmol) in EtOH (10 mL) and aq 1 M NaOH
(10 mL) was stirred at 80 °C for 4 h. The
solution was concentrated in vacuo, diluted
with EtOAc, washed with water and brine,
dried over Na2SO4, filtered and concentrated
to give an amine, which was dissolved in
CH2Cl2 (10 mL). To this solution were added
satd aq NaHCO3 (10 mL) and chloroformic
acid 2,2,2-trichloroethyl ester (1.60 g, 7.55
mmol) with stirring at rt. After 1 h, the mix-
ture was diluted with CH2Cl2, washed with
water and brine, dried over MgSO4, filtered,
concentrated in vacuo, and chromatographed
on a silica-gel column. Elution with 4:1 hex-
ane–EtOAc gave 50 (4.54 g, 87%) as a gum.
Allyl 6-O-benzyloxycarbonyl-2-deoxy-3-O-
[(R) - 3 - (dodecyloxy)tetradecyl] - 2 - [(2,2,2-tri-
chloroethoxycarbonyl)amino]-h- -glucopyran-
D
oside (52).—To a solution of 51 (526 mg,
0.678 mmol) in CH2Cl2 (5 mL) were added
pyridine (332 mg, 4.20 mmol) and benzyloxy-
carbonyl chloride (0.50 mL, 3.50 mmol) at
0 °C under N2. After stirring for 3 h at rt, the
solution was diluted with EtOAc, washed with
aq 1 M NH4Cl, satd aq NaHCO3, and brine,
dried over MgSO4, and filtered. The filtrate
was concentrated in vacuo, and chro-
matographed on a silica-gel column. Elution
with 3:1 hexane–EtOAc gave 52 (581 mg,
94%) as a gum. IR wmax(KBr) 3522, 3329,
IR wmax(CHCl3) 3439, 2928, 2855, 1743 cm−1
.
1H NMR (400 MHz, CDCl3) l 0.88 (6 H, t, J
6.6–7.3 Hz), 1.21–1.68 (48 H, m, containing 3
H, s, at l 1.41, and 3 H, s, at l 1.50),
3.30–3.48 (4 H, m), 3.57–3.77 (4 H, m), 3.82–
3.92 (3 H, m), 3.98 (1 H, dd, J 6.6, 12.5 Hz),
1
2923, 2853, 1725, 1709 cm−1. H NMR (400
MHz, CDCl3) l 0.88 (6 H, t, J 6.6 Hz),

224
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
tetradecyl]-2-[(2,2,2-trichloroethoxycarbonyl)-
amino]-
1.59–1.21 (40 H, m), 1.71–1.74 (2 H, m),
3.33–3.48 (4 H, m), 3.56 (1 H, m), 3.64 (1 H,
bs, OH), 3.69 (1 H, m), 3.79–3.84 (2 H, m),
3.88 (1 H, dt, J 3.7, 10.3 Hz), 3.96 (1 H, dd, J
6.6, 12.5 Hz), 4.15 (1 H, dd, J 5.1, 12.5 Hz),
4.39–4.46 (2 H, m), 4.68, 4.81 (2 H, AB-q, J
12.5 Hz), 4.85 (1 H, d, J 3.7 Hz), 5.18 (2 H, s),
5.20–5.30 (3 H, m, containing NH), 5.87 (1 H,
m), 7.32–7.40 (5 H, m). FABMS (positive-
ion): m/z 908 [M+H]+. HRFABMS (posi-
tive-ion): m/z Calcd for C46H77Cl3NO10:
908.4613; Found: 908.4592. Anal. Calcd for
C46H76Cl3NO10 (909.5): C, 60.75; H, 8.42; Cl,
11.70; N, 1.54. Found: C, 60.44; H, 8.17; Cl,
11.63; N, 1.56.
D
-glucopyranose (54).—To a solution
of 53 (613 mg, 0.537 mmol) in THF (6 mL)
was added bis(methyldiphenylphosphine)-
cyclooctadieneiridium(I)
hexafluorophos-
phate, [C8H12Ir(PMePh2)2]PF6 (31.5 mg). The
air in the reaction flask was completely re-
placed with N2 and then further replaced with
hydrogen to activate the iridium complex. Im-
mediately after 1 min, when the red color
solution of the iridium complex had become
almost colorless, the hydrogen was completely
replaced with N2. This solution was stirred for
3 h at rt. After confirming a double bond shift
to an enol ether from the 1-allyloxy group (as
indicated by a slightly higher Rf value on the
TLC), water (5 mL) and I2 (276 mg) were
added to this solution. After stirring for 1 h at
rt, the mixture was diluted with EtOAc,
washed with 10% aq Na2S2O3, satd aq
NaHCO3, and brine, dried over MgSO4, and
concentrated to give a mixture that was chro-
matographed on a silica-gel column. Elution
with 3:2 hexane–EtOAc gave 54 (421 mg,
71%) as a gum. IR wmax(CHCl3) 3600, 3435,
Allyl 6-O-benzyloxycarbonyl-2-deoxy-4-O-
(diphenylphosphono)-3-O-[(R)-3-(dodecyloxy)-
tetradecyl]-2-[(2,2,2-trichloroethoxycarbonyl)-
amino]-h- -glucopyranoside (53).—To a solu-
D
tion of 52 (553 mg, 0.608 mmol) and DMAP
(111 mg, 0.911 mmol) in CH2Cl2 (5 mL) was
added dropwise diphenyl chlorophosphate
(0.19 mL, 0.917 mmol). After stirring for 2 h
at rt, the reaction mixture was diluted with
EtOAc, washed with satd aq NaHCO3 and
brine, dried over MgSO4, filtered, and concen-
trated in vacuo to give a residue, which was
chromatographed on a silica-gel column. Elu-
tion with 3:1 hexane–EtOAc gave 53 (663 mg,
96%) as a gum. IR wmax(CHCl3) 3436, 2928,
1
2928, 2855, 1747 cm−1. H NMR (400 MHz,
CDCl3) l 0.88 (6 H, t, J 6.6 Hz), 1.18–1.30
(38 H, m), 1.43–1.45 (2 H, m), 1.62–1.65 (2
H, m), 3.20–3.27 (3 H, m, containing 2 H, t,
J 6.6 Hz at l 3.25), 3.41 (1 H, bs, OH),
3.64–3.84 (3 H, m), 3.94 (1 H, m), 4.19–4.39
(3 H, m), 4.62 (1 H, q, J 9.5 Hz), 4.69, 4.76 (2
H, AB-q, J 12.1 Hz), 5.06, 5.11 (2 H, AB-q, J
11.7 Hz), 5.25 (1 H, t, J 3.7 Hz), 5.43 (1 H, d,
J 9.5 Hz, NH), 7.12–7.34 (15 H, m). FABMS
(positive-ion): m/z 1100 [M+H]+. HR-
FABMS (positive-ion): m/z Calcd for
2855, 1747 cm−1
.
1H NMR (400 MHz,
CDCl3) l 0.88 (6 H, t, J 6.6 Hz), 1.18–1.30
(38 H, m), 1.43–1.45 (2 H, m), 1.61–1.63 (2
H, m), 3.20 (1 H, m), 3.25 (2 H, t, J 6.6 Hz),
3.64–3.72 (2 H, m), 3.80 (1 H, m), 3.95–4.02
(3 H, m), 4.14 (1 H, dd, J 5.1, 13.2 Hz), 4.29
(1 H, dd, J 5.1, 11.7 Hz), 4.36 (1 H, dd, J 1.5,
11.7 Hz), 4.63 (1 H, q, J 9.5 Hz), 4.67, 4.79 (2
H, AB-q, J 12.5 Hz), 4.89 (1 H, d, J 3.7 Hz),
5.06, 5.11 (2 H, AB-q, J 12.5 Hz), 5.21–5.30
(3 H, m, containing NH), 5.87 (1 H, m),
7.12–7.36 (15 H, m). FABMS (positive-ion):
m/z 1162 [M+Na]+; 1140 [M+H]+. HR-
FABMS (positive-ion): m/z Calcd for
C55H82Cl3NO13P:
1100.4601.
1100.4589;
Found:
Allyl 6-O-(tert-butyldimethylsilyl)-2-deoxy-
3-O-[(R)-3-(dodecyloxy)tetradecyl]-2-[(2,2,2-
trichloroethoxycarbonyl)amino] - h -
D
- gluco-
pyranoside (55).—To a solution of 51 (1.32 g,
1.70 mmol) in CH2Cl2 (6 mL) were added
DMAP (332 mg, 2.56 mmol) and tert-
butyldimethylsilyl chloride (385 mg, 2.56
mmol). After stirring for 3 h at rt, the mixture
was diluted with EtOAc, washed with water,
satd aq NaHCO3, and brine, dried over
MgSO4, and filtered. The filtrate was concen-
trated in vacuo and chromatographed on a
C58H86Cl3NO13P:
1140.4902;
Found:
1140.4890. Anal. Calcd for C58H85Cl3NO13P
(1141.6): C, 61.02; H, 7.51; Cl, 9.32; N, 1.23;
P, 2.71. Found: C, 60.69; H, 7.61; Cl, 9.44; N,
1.19; P, 2.96.
6-O-Benzyloxycarbonyl-2-deoxy-4-O-(di-
phenylphosphono) - 3 - O - [(R) - 3 - (dodecyloxy)-

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
225
1
3499, 3373, 2921, 2851, 1713, 1646 cm−1. H
NMR (400 MHz, CDCl3) l 0.88 (6 H, t, J 6.6
Hz), 1.18–1.47 (40 H, m), 1.65 (1 H, q, J 6.6
Hz), 3.21–3.29 (3 H, m), 3.59–3.70 (5 H, m),
3.88 (1 H, m), 3.97–4.03 (2 H, m), 4.17 (1 H,
dd, J 5.9, 13.2 Hz), 4.65 (1 H, q, J 9.5 Hz),
4.72, 4.76 (2 H, AB-q, J 12.5 Hz), 4.93 (1 H,
d, J 3.7 Hz), 5.23–5.33 (3 H, m, containing
NH), 5.89 (1 H, m), 7.18–7.37 (10 H, m).
FABMS (positive-ion): m/z 1006 [M+H]+.
HRFABMS (positive-ion): m/z Calcd for
silica-gel column. Elution with 9:1 hexane–
EtOAc gave 55 (1.49 g, 99%) as a gum. IR
1
wmax(CHCl3) 3438, 2928, 2856, 1742 cm−1. H
NMR (400 MHz, CDCl3) l 0.09 (6 H, s),
0.86–0.91 (15 H, m), 1.21–1.62 (40 H, m),
1.72 (2 H, q, J 5.9 Hz), 3.36–3.67 (7 H, m,
containing OH), 3.76–3.89 (5 H, m), 3.98 (1
H, dd, J 6.6, 12.5 Hz), 4.19 (1 H, dd, J 5.1,
12.5 Hz), 4.70, 4.79 (2 H, AB-q, J 12.5 Hz),
4.86 (1 H, d, J 3.7 Hz), 5.20–5.31 (3 H, m,
containing NH), 5.90 (1 H, m). FABMS (pos-
itive-ion): m/z 910 [M+Na]+; 888 [M+H]+.
HRFABMS (positive-ion): m/z Calcd for
C50H80Cl3NO11P:
1006.4535;
Found:
1006.4539. Anal. Calcd for C50H79Cl3NO11P
(1007.5): C, 59.61; H, 7.90; Cl, 10.56; N, 1.39;
P, 3.07. Found: C, 59.29; H, 7.82; Cl, 10.66;
N, 1.37; P, 3.09.
C44H84Cl3NNaO8Si:
910.4930;
Found:
910.4911. Anal. Calcd for C44H84Cl3NO8Si
(889.6): C, 59.41; H, 9.52; Cl, 11.96; N, 1.58.
Found: C, 59.13; H, 9.30; Cl, 12.02; N, 1.49.
Allyl 2-deoxy-4-O-(diphenylphosphono)-3-
O-[(R)-3-(dodecyloxy)tetradecyl]-6-O-methyl-
Allyl
6-O-tert-butyldimethylsilyl-2-deoxy-
4-O-(diphenylphosphono)-3-O-[(R)-3-(dode-
cyloxy)tetradecyl] - 2 - [(2,2,2 - trichloroethoxy-
2-[(2,2,2-trichloroethoxycarbonyl)amino]-h- -
D
glucopyranoside (58).—To a solution of 57
(385 mg, 0.382 mmol) in CH2Cl2 (5 mL) were
added 2,6-di-tert-butyl-4-methylpyridine (91
mg, 0.441 mmol) and trimethyloxonium tetra-
fluoroborate (64 mg, 0.435 mmol) at rt. After
3 h, the reaction mixture was diluted with
EtOAc, washed with satd aq NaHCO3 and
brine, dried over MgSO4, filtered, concen-
trated, and chromatographed on a silica-gel
column. Elution with 7:3 hexane–EtOAc gave
58 (357 mg, 91%) as a gum. IR wmax(CHCl3)
carbonyl)amino]-h- -glucopyranoside (56).—
D
Compound 55 (1.45 g, 1.63 mmol) was treated
as described in the formation of 53 from 52 to
give 56 (1.81 g, 99%) as a gum. IR
1
wmax(CHCl3) 3436, 2928, 2856, 1744 cm−1. H
NMR (400 MHz, CDCl3) l 0.01 (6 H, s),
0.91–0.85 (15 H, m), 1.18–1.65 (42 H, m),
3.19–3.26 (3 H, m), 3.65–3.83 (6 H, m), 3.86–
4.03 (2 H, m), 4.22 (1 H, dd, J 5.9, 13.2 Hz),
4.55 (1 H, q, J 9.5 Hz), 4.69, 4.81 (2 H, AB-q,
J 12.1 Hz), 4.90 (1 H, d, J 3.7 Hz), 5.23–5.33
(3 H, m, containing NH), 5.92 (1 H, m),
7.15–7.35 (10 H, m). FABMS (positive-ion):
m/z 1120 [M+H]+. HRFABMS (positive-
ion): m/z Calcd for C56H94Cl3NO11PSi:
1120.5399; Found: 1120.5409. Anal. Calcd for
C56H93Cl3NO11PSi (1121.8): C, 59.96; H, 8.36;
Cl, 9.48; N, 1.25; P, 2.76. Found: C, 59.59; H,
8.23; Cl, 9.66; N, 1.18; P, 2.81.
3436, 2928, 2855, 1744 cm−1. H NMR (400
1
MHz, CDCl3) l 0.88 (6 H, t, J 6.6 Hz),
1.17–1.32 (38 H, m), 1.43–1.44 (2 H, m),
1.62–1.64 (2 H, m), 3.19–3.26 (6 H, m, con-
taining 3 H, s, at l 3.24), 3.48 (1 H, dd, J 2.2,
11.0 Hz), 3.54 (1 H, dd, J 4.4, 11.0 Hz),
3.65–3.89 (4 H, m), 3.98–4.04 (2 H, m), 4.21
(1 H, dd, J 5.1, 13.2 Hz), 4.67 (1 H, m), 4.69,
4.77 (2 H, AB-q, J 11.7 Hz), 4.93 (1 H, d, J
3.7 Hz), 5.23–5.33 (3 H, m, containing NH),
5.90 (1 H, m), 7.15–7.35 (10 H, m). FABMS
(positive-ion): m/z 1020 [M+H]+. HR-
FABMS (positive-ion): m/z Calcd for
Allyl 2-deoxy-4-O-(diphenylphosphono)-3-
O-[(R)-3-(dodecyloxy)tetradecyl]-2-[(2,2,2-tri-
chloroethoxycarbonyl)amino]-h- -glucopyran-
D
oside (57).—To a solution of 56 (1.65 g, 1.47
mmol) in THF (9 mL) was added aq 3 M HCl
(1.5 mL) with stirring. After 3 h at rt, the
reaction mixture was diluted with EtOAc,
washed with satd aq NaHCO3 and brine,
dried over MgSO4, filtered, concentrated, and
chromatographed on a silica-gel column. Elu-
tion with 3:2 hexane–EtOAc gave 57 (1.31 g,
89%) as an amorphous solid. IR wmax(KBr)
C51H82Cl3NO11P:
1020.4691;
Found:
1020.4669. Anal. Calcd for C51H81Cl3NO11P
(1021.5): C, 59.97; H, 7.99; Cl, 10.41; N, 1.37;
P, 3.03. Found: C, 59.50; H, 7.85; Cl, 10.51;
N, 1.29; P, 3.27.
2 - Deoxy - 4 - O - (diphenylphosphono) - 3 - O-
[(R)-3-(dodecyloxy)tetradecyl]-6-O-methyl-2-
[(2,2,2 - trichloroethoxycarbonyl)amino] -
D
-

226
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
Cl3FNO10P (1009.5): C, 59.49; H, 7.79; Cl,
10.54; F, 1.88; N, 1.39; P, 3.07. Found: C,
59.45; H, 7.68; Cl, 10.45; F, 2.15; N, 1.48; P,
3.24.
glucopyranose (59).—Compound 58 (695 mg,
0.681 mmol) was treated as described in the
formation of 54 from 53 to give 59 (483 mg,
72%) as a gum. IR wmax(KBr) 3426, 3339,
1
2922, 2851, 1721 cm−1. H NMR (400 MHz,
2,6-Dideoxy-4-O-(diphenylphosphono)-3-O-
[(R) - 3 - (dodecyloxy)tetradecyl] - 6 - fluoro - 2-
CDCl3) l 0.88 (6 H, t, J 6.6 Hz), 1.17–1.32
(38 H, m), 1.42–1.44 (2 H, m), 1.64–1.65 (2
H, m), 3.16–3.26 (6 H, m, containing 3 H, s,
at l 3.21 and 2 H, t, J 6.6 Hz, at l 3.25),
3.43–3.51 (2 H, m), 3.65–3.85 (4 H, m, con-
taining OH), 3.95 (1 H, m), 4.13 (1 H, m),
4.55 (1 H, q, J 9.5 Hz), 4.70, 4.75 (2 H, AB-q,
J 11.7 Hz), 5.27 (1 H, t, J 3.7 Hz), 5.40 (1 H,
d, J 9.5 Hz, NH), 7.16–7.35 (10 H, m).
FABMS (positive-ion): m/z 1018 [M+K]+,
1002 [M+Na]+, 980 [M+H]+. HRFABMS
(positive-ion): m/z Calcd for C48H78Cl3NO11P:
980.4360; Found: 980.4378. Anal. Calcd for
C48H77Cl3NO11P (981.5): C, 58.74; H, 7.91; Cl,
10.84; N, 1.43; P, 3.16. Found: C, 58.49; H,
8.15; Cl, 10.74; N, 1.41; P, 3.14.
[(2,2,2 - trichloroethoxycarbonyl)amino] -
D
-
glucopyranose (61).—Compound 60 (416 mg,
0.412 mmol) was treated as described in the
formation of 54 from 53 to give 61 (329 mg,
82%) as a gum. IR wmax(CHCl3) 3602, 3435,
1
2928, 2855, 1745 cm−1. H NMR (400 MHz,
CDCl3) l 0.88 (6 H, t, J 6.6 Hz), 1.18–1.32
(38 H, m), 1.44–1.45(2 H, m), 1.62–1.67 (2 H,
m), 3.21–3.27 (3 H, m, containing 2 H, t, J 6.6
Hz, at l 3.26), 3.65–3.85 (4 H, m, containing
OH), 3.96 (1 H, m), 4.17 (1 H, m), 4.44 (1 H,
m), 4.56 (1 H, m), 4.60 (1 H, q, J 9.5 Hz),
4.71, 4.76 (2 H, AB-q, J 11.7 Hz), 5.31 (1 H,
t, J 3.7 Hz), 5.47 (1 H, d, J 9.5 Hz, NH),
7.17–7.35 (10 H, m). FABMS (positive-ion):
m/z 968 [M+H]+. HRFABMS (positive-ion):
m/z Calcd for C47H75Cl3FNO10P: 968.4187;
Found: 968.4178. Anal. Calcd for C47H74Cl3-
FNO10P (969.4): C, 58.23; H, 7.69; Cl, 10.97;
F, 1.96; N, 1.45; P, 3.20. Found: C, 58.27; H,
7.76; Cl, 11.20; F, 2.04; N, 1.43; P, 3.25.
Allyl 2,6-dideoxy-4-O-(diphenylphosphono)-
3-O-[(R)-3-(dodecyloxy)tetradecyl]-6-fluoro-
2-[(2,2,2-trichloroethoxycarbonyl)amino]-h- -
D
glucopyranoside (60).—To a solution of 57
(521 mg, 0.517 mmol) in DME (5 mL) was
gradually added a solution of DAST (0.2 mL,
1.51 mmol) in DME (1 mL) at −40 °C with
stirring under N2. After stirring for 30 min at
−40 °C, followed by 2 h at rt, water was
added to the reaction mixture, and it was
extracted with EtOAc. The organic layer was
washed with satd aq NaHCO3 and brine,
dried over MgSO4, and concentrated to give a
crude oil. The crude product was chro-
matographed on a silica-gel column. Elution
with 3:1 hexane–EtOAc gave 60 (432 mg,
83%). IR wmax(CHCl3) 3691, 3436, 2928, 2855,
Diphenylmethyl 2,6-anhydro-7-O-[6-O-ben-
zyloxycarbonyl - 2 - deoxy - 4 - O - (diphenylphos-
phono) - 3 - O - [(R) - 3 - (dodecyloxy)tetradecyl]-
2-[(2,2,2-trichloroethoxycarbonyl)amino]-i- -
D
glucopyranosyl] - 3 - [(R) - 3 - (benzyloxy)tetra-
decanamido] - 4 - O - [(R) - 3 - (benzyloxy)tetra-
decyl] - 3 - deoxy -
D
- glycero -
D
- ido - heptonate
(62).—To a solution of 54 (521 mg, 0.473
mmol) in CH2Cl2 (10 mL) were added
Cl3CCN (0.70 mL, 6.98 mmol) and a catalytic
amount of Cs2CO3 (77 mg, 0.237 mmol). After
stirring for 1 h at rt, the reaction mixture was
quenched with satd aq NaHCO3 (20 mL), and
extracted with CH2Cl2. The extract was
washed with brine and dried over Na2SO4.
Removal of the solvent in vacuo gave a crude
imidate (591 mg), which was immediately used
for subsequent glycosylation without further
purification. In a N2 atmosphere, a solution of
the thus-obtained imidate (591 mg), diol 1
(234 mg, 0.236 mmol), and MS4A (750 mg) in
CH2Cl2 (10 mL) was stirred at rt. After stir-
ring for 1 h, a catalytic amount of TMSOTf
(15 mL, 0.083 mmol) was added to the mix-
ture at −40 °C. After stirring for 2 h at
1
1744 cm−1. H NMR (400 MHz, CDCl3) l
0.88 (6 H, t, J 6.6–7.3 Hz), 1.17–1.32 (38 H,
m), 1.43–1.45 (2 H, m), 1.57–1.65 (2 H, m),
3.21 (1 H, m), 3.25 (2 H, t, J 6.6–7.3 Hz),
3.65–3.83 (3 H, m), 3.90–4.05 (3 H, m), 4.20
(1 H, dd, J 5.9, 13.2 Hz), 4.46 (1 H, m),
4.55–4.64 (2 H, m), 4.68, 4.79 (2 H, AB-q, J
11.7 Hz), 4.94 (1 H, d, J 3.7 Hz), 5.24–5.34 (3
H, m, containing NH), 5.89 (1 H, m), 7.16–
7.34 (10 H, m). FABMS (positive-ion): m/z
1008 [M+H]+. HRFABMS (positive-ion): m/
z Calcd for C50H79Cl3FNO10P: 1008.4491;
Found: 1008.4485. Anal. Calcd for C50H78-

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
227
NaO19P: 1976.0629; Found: 1976.0585. Anal.
Calcd for C110H164Cl3N2O19P (1955.8): C,
67.55; H, 8.45; Cl, 5.44; N, 1.43; P, 1.58.
Found: C, 67.53; H, 8.25; Cl, 5.14; N, 1.43; P,
1.32.
−40 °C, the mixture was quenched with satd
aq NaHCO3, diluted with EtOAc, washed
with water and brine, dried over MgSO4,
filtered, and concentrated in vacuo to give a
mixture, which was chromatographed on a
silica-gel column. Elution with 7:3 hexane–
EtOAc gave 62 (365 mg, 75%) as a gum. IR
wmax(CHCl3) 3691, 3604, 3519, 3405, 2928,
Diphenylmethyl
2,6-anhydro-7-O-[2,6-
dideoxy-4-O-(diphenylphosphono)-3-O-[(R)-3-
(dodecyloxy)tetradecyl]-6-fluoro-2-[(2,2,2-tri-
1
2855, 1735, 1669 cm−1. H NMR (400 MHz,
chloroethoxycarbonyl)amino]-i-
osyl]-3-[(R)-3-(benzyloxy)tetradecanamido]-4-
O-[(R)-3-(benzyloxy)tetradecyl]-3-deoxy-
glycero- -ido-heptonate (64).—Compound 61
D
-glucopyran-
CDCl3) l 0.88 (12 H, t, J 6.6 Hz), 1.24–1.74
(84 H, m), 2.24 (2 H, d, J 5.9 Hz), 3.02 (1 H,
bs, OH), 3.14–3.27 (4 H, m), 3.43–3.72 (11 H,
m), 3.92–4.07 (2 H, m), 4.21–4.45 (7 H, m,
containing 2 H, AB-q, J 11.5 Hz, at l 4.39,
4.44, and 2 H, AB-q, J 11.0 Hz, at l 4.28,
4.33), 4.51 (1 H, q, J 9.5 Hz), 4.61 (1 H, d, J
5.9 Hz), 4.66, 4.71 (2 H, AB-q, J 11.7 Hz),
4.86 (1 H, m), 5.04, 5.10 (2 H, AB-q, J 11.7
Hz), 5.39 (1 H, m, NH), 6.81 (1 H, m, NH),
6.83 (1 H, s), 7.11–7.35 (35 H, m). FABMS
(positive-ion): m/z 2095 [M+Na]+. HR-
FABMS (positive-ion): m/z Calcd for
C117H168Cl3N2O21PNa: 2096.0841; Found:
2096.0845. Anal. Calcd for C117H168Cl3N2O21P
(2075.9): C, 67.69; H, 8.16; Cl, 5.12; N, 1.35;
P, 1.49. Found: C, 67.21; H, 8.04; Cl, 5.27; N,
1.34; P, 1.37.
D
-
D
(230 mg, 0.238 mmol) was treated as described
in the formation of 62 from 54 to give 64 (185
mg, 80%) as a gum. IR wmax(CHCl3) 2928,
1
2855, 1735, 1669 cm−1. H NMR (400 MHz,
CDCl3) l 0.88 (12 H, t, J 6.6 Hz), 1.25–1.74
(84 H, m), 2.25 (2 H, d, J 5.9 Hz), 3.03 (1 H,
bs, OH), 3.15–3.27 (4 H, m), 3.43–3.75 (11 H,
m), 3.92–4.02 (2 H, m), 4.26–4.57 (8 H, m,
containing 2 H, AB-q, J 11.4 Hz, at l 4.28,
4.33, and 2 H, AB-q, J 11.7 Hz, at l 4.40 and
4.45), 4.61 (1 H, d, J 5.1 Hz), 4.67, 4.75 (2 H,
AB-q, J 11.7 Hz), 4.90 (1 H, m), 5.43 (1 H, m,
NH), 6.83 (1 H, s), 6.82 (1 H, m, NH),
7.17–7.35 (30 H, m). FABMS (positive-ion):
m/z 1963 [M+Na]+. HRFABMS (positive-
ion): m/z Calcd for C109H161Cl3FN2NaO18P:
1964.0429; Found: 1964.0414. Anal. Calcd for
C109H161Cl3FN2O18P (1943.78): C, 67.35; H,
8.35; Cl, 5.47; F, 0.98; N, 1.44; P, 1.59.
Found: C, 67.34; H, 8.10; Cl, 5.39; F, 1.26; N,
1.48; P, 1.43.
Diphenylmethyl 2,6-anhydro-7-O-[2-deoxy-
4-O-(diphenylphosphono)-3-O-[(R)-3-(dode-
cyloxy)tetradecyl] - 6 - O - methyl - 2 - [(2,2,2 - tri-
chloroethoxycarbonyl)amino]-i-
osyl]-3-[(R)-3-(benzyloxy)tetradecanamido]-4-
O-[(R)-3-(benzyloxy)tetradecyl]-3-deoxy-
glycero- -ido-heptonate (63).—Compound 59
D
-glucopyran-
D
-
D
Diphenylmethyl 2,6-anhydro-7-O-[2-acetam-
ido-6-O-(benzyloxycarbonyl)-2-deoxy-4-O-
(diphenylphosphono)-3-O-[(R)-3-(dodecyloxy)-
(438 mg, 0.446 mmol) was treated as described
in the formation of 62 from 54 to give 63 (298
mg, 68%) as a gum. IR wmax(CHCl3) 2928,
tetradecyl] - i -
(benzyloxy)tetradecanamido] - 4 - O - [(R) - 3-
(benzyloxy)tetradecyl]-3-deoxy- -glycero- -
D
- glucopyranosyl] - 3 - [(R) - 3-
1
2855, 1735, 1668 cm−1. H NMR (400 MHz,
CDCl3) l 0.88 (12 H, t, J 6.6 Hz), 1.24–1.75
(84 H, m), 2.23 (2 H, d, J 5.9 Hz), 3.03 (1 H,
bs, OH), 3.10–3.27 (7 H, m, containing 3 H, s,
at l 3.21), 3.41–3.76 (13 H, m), 3.89–4.00 (2
H, m), 4.26–4.34 (3 H, m, containing 2 H,
AB-q, J 11.4 Hz, at l 4.28 and 4.33), 4.40,
4.44 (2 H, AB-q, J 11.7 Hz), 4.54 (1 H, q, J
9.5 Hz), 4.61 (1 H, d, J 5.9 Hz), 4.67, 4.75 (2
H, AB-q, J 11.7 Hz), 4.84 (1 H, m), 5.33 (1 H,
m, NH), 6.79 (1 H, d, J 9.5 Hz, NH), 6.83 (1
H, s), 7.15–7.35 (30 H, m). FABMS (positive-
ion): m/z 1975 [M+Na]+. HRFABMS (posi-
tive-ion): m/z Calcd for C110H164Cl3N2-
D
D
ido-heptonate (65).—To a solution of 62 (230
mg, 0.111 mmol) in AcOH (5 mL) was added
activated zinc dust (148 mg, 2.27 mmol). After
stirring vigorously for 5 h at rt, the solution
was filtered to remove the zinc dust and con-
centrated in vacuo to give a crude product.
The product was diluted with EtOAc, washed
with satd aq NaHCO3
and brine, dried over
Na2SO4, filtered, and concentrated in vacuo.
The crude product was then dissolved in THF
(3 mL) and water (2 mL), and pyridine (45
mL) and Ac2O (55 mL) were added. After

228
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
Diphenylmethyl 2,6-anhydro-7-O-[2-acetam-
ido-2,6-dideoxy-4-O-(diphenylphosphono)-3-
O-[(R)-3-(dodecyloxy)tetradecyl]-6-fluoro-i-
stirring for 2 h at rt, the mixture was diluted
with EtOAc, washed with water and brine,
dried over MgSO4, filtered, and concentrated
in vacuo, and chromatographed on a silica-gel
column. Elution with 3:2 hexane–EtOAc gave
65 (152 mg, two steps, 71%) as a gum. IR
wmax(CHCl3) 3436, 2928, 2855, 1745, 1670
cm−1. 1H NMR (400 MHz, CDCl3) l 0.89 (12
H, t, J 6.6 Hz), 1.26–1.73 (84 H, m), 1.91 (3
H, s), 2.25 (2 H, d, J 5.1 Hz), 3.03 (1 H, m),
3.27–3.72 (14 H, m, containing OH), 4.00–
4.07 (2 H, m), 4.23 (1 H, dd, J 5.1, 11.7 Hz),
4.28–4.39 (5 H, m), 4.50 (1 H, q, J 9.5 Hz),
4.43 (2 H, s), 4.62 (1 H, d, J 5.1 Hz), 5.05,
5.11 (2 H, AB-q, J 11.7 Hz), 5.21 (1 H, d, J
8.1 Hz), 6.07 (1 H, d, J 6.6 Hz, NH), 6.80 (1
H, d, J 8.8 Hz, NH), 6.82 (1 H, s), 7.12–7.33
(35 H, m). FABMS (positive-ion): m/z 1963
[M+Na]+. HRFABMS (positive-ion): m/z
Calcd for C116H169N2NaO20P: 1964.1904;
D
-glucopyranosyl]-3-[(R)-3-(benzyloxy)tetra-
decanamido] - 4 - O - [(R) - 3 - (benzyloxy)tetra-
decyl] - 3 - deoxy - - glycero - - ido - heptonate
D
D
(67).—Compound 64 (180 mg, 0.093 mmol)
was treated as described in the formation of
65 from 62 to give 67 (115 mg, 68%) as a gum.
IR wmax(CHCl3) 3529, 3335, 3064, 3032, 2924,
1
2854, 1733, 1646 cm−1. H NMR (400 MHz,
CDCl3) l 0.88 (12 H, t, J 6.6 Hz), 1.25–1.74
(84 H, m), 1.91 (3 H, s), 2.24 (2 H, d, J 5.9
Hz), 3.04 (1 H, m), 3.22–3.33 (4 H, m, con-
taining OH), 3.41–3.71 (11 H, m), 4.03–4.11
(2 H, m), 4.27–4.57 (8 H, m, containing 2 H,
AB-q, J 11.4 Hz, at l 4.29, 4.34, and 2 H, s,
at l 4.42), 4.61 (1 H, d, J 5.1 Hz), 5.21 (1 H,
d, J 8.1 Hz), 6.03 (1 H, d, J 6.6 Hz, NH), 6.80
(1 H, d, J 11.7 Hz, NH), 6.82 (1 H, s),
7.17–7.37 (30 H, m). FABMS (positive-ion):
m/z 1831 [M+Na]+. HRFABMS (positive-
ion): m/z Calcd for C108H162FN2NaO17P:
1832.1493; Found: 1832.1459. Anal. Calcd for
C108H162FN2O17P (1810.4): C, 71.65; H, 9.02;
F, 1.05; N, 1.55; P, 1.71. Found: C, 71.34; H,
8.88; F, 1.18; N, 1.50; P, 1.52.
Found:
1964.1870.
Anal.
Calcd
for
C116H169N2O20P (1942.6): C, 71.72; H, 8.77; N,
1.44; P, 1.59. Found: C, 71.46; H, 8.60; N,
1.42; P, 1.49.
Diphenylmethyl 2,6-anhydro-7-O-[2-acetam-
ido-2-deoxy-4-O-(diphenylphosphono)-3-O-
[(R)-3-(dodecyloxy)tetradecyl]-6-O-methyl-i-
D
-glucopyranosyl]-3-[(R)-3-(benzyloxy)tetra-
decanamido] - 4 - O - [(R) - 3 - (benzyloxy)tetra-
decyl] - 3 - deoxy - - glycero - - ido - heptonate
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-4-O-
(diphenylphosphono)-3-O-[(R)-3-(dodecyloxy)-
D
D
tetradecyl] - i -
(hydroxy)tetradecanamido]-4-O-[(R)-3-(hydro-
xy)tetradecyl]-3-deoxy- -glycero- -ido-hep-
D
- glucopyranosyl] - 3 - [(R) - 3-
(66).—Compound 63 (229 mg, 0.117 mmol)
was treated as described in the formation of
65 from 62 to give 66 (158 mg, 74%) as a gum.
IR wmax (CHCl3) 3436, 2928, 2855, 1732, 1670
cm−1. 1H NMR (400 MHz, CDCl3) l 0.88 (12
H, t, J 6.6 Hz), 1.25–1.74 (84 H, m), 1.91 (3
H, s), 2.23 (2 H, d, J 5.1 Hz), 3.11 (1 H, m),
3.22–3.30 (6 H, m, containing 3 H, s, at l
3.22), 3.40–3.74 (14 H, m, containing OH),
3.99–4.05 (2 H, m), 4.29, 4.33 (2 H, AB-q, J
11.4 Hz), 4.35 (1 H, m), 4.42 (2 H, s), 4.52 (1
H, q, J 9.5 Hz), 4.61 (1 H, d, J 5.9 Hz), 5.15
(1 H, d, J 8.1 Hz), 5.97 (1 H, d, J 7.3 Hz,
NH), 6.78 (1 H, d, J 8.8 Hz, NH), 6.81 (1 H,
s), 7.15–7.33 (30 H, m). FABMS (positive-
ion): m/z 1843 [M+Na]+. HRFABMS (posi-
tive-ion): m/z Calcd for C109H165N2NaO18P:
1844.1693; Found: 1844.1700. Anal. Calcd for
C109H165N2O18P (1822.5): C, 71.84; H, 9.13; N,
1.54; P, 1.70. Found: C, 71.38; H, 8.96; N,
1.51; P, 1.53.
D
D
tonic acid (68).—A solution of 65 (115 mg,
0.059 mmol) in EtOH (5 mL) containing 20%
Pd(OH)2–C (111 mg) was stirred vigorously
under H2 for 18 h at rt. The reaction mixture
was filtered and concentrated in vacuo to give
a crude product. The crude product was
purified by preparative silica gel thin-layer
chromatography (8:1 CHCl3–MeOH) to give
68 (62.3 mg, 72%) as an amorphous solid. IR
wmax(KBr) 3320 (broad), 3072, 2924, 2854,
1726, 1656 cm−1
.
1H NMR (400 MHz,
CD3OD) l 0.90 (12 H, t, J 6.6 Hz), 1.20–1.76
(84 H, m), 2.30–2.41 (2 H, m), 2.00 (3 H, s),
3.25–3.35 (3 H, m), 3.46 (1 H, t, J 8.1 Hz),
3.52–3.93 (14 H, m), 4.04 (1 H, m), 4.21 (1 H,
m), 4.48 (1 H, d, J 5.1 Hz), 4.53 (1 H, m), 4.63
(1 H, m), 7.19–7.40 (10 H, m). FABMS (pos-
itive-ion): m/z 1483 [M+Na]+, 1461 [M+
H]+. HRFABMS (positive-ion): m/z Calcd

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
229
PtO2 (55.6 mg) was stirred vigorously under
H2 for 5 h at rt. The reaction mixture was
filtered and concentrated in vacuo to give a
residue. The residue was dissolved in CHCl3 (5
mL), MeOH (10 mL) and aq 0.1 M HCl (4
mL). To this solution was added another vol-
ume of CHCl3 (5 mL) and 0.1 M HCl (5 mL)
to separate the solution into two phases. The
lower CHCl3 phase was collected and concen-
trated to give 71 (48.3 mg, 96%) as a white
powder, mp 179.0–181.0 °C. [h]2D4 −25.5° (c
0.78, CHCl3). IR wmax(KBr) 3294 (broad),
for C81H141N2NaO18P: 1483.9789; Found:
1483.9815.
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-4-O-
(diphenylphosphono)-3-O-[(R)-3-(dodecyloxy)-
tetradecyl]-6-O-methyl-i-
[(R)-3-(hydroxy)tetradecanamido]-4-O-[(R)-3-
(hydroxy)tetradecyl]-3-deoxy- -glycero- -ido-
D
-glucopyranosyl]-3-
D
D
heptonic acid (69).—Compound 66 (136 mg,
0.074 mmol) was treated as described in the
formation of 68 from 65 to give 69 (98.2 mg,
89%) as an amorphous solid. IR wmax(KBr)
3318 (broad), 3072, 2924, 2854, 1729, 1656
cm−1. H NMR (400 MHz, CD3OD) l 0.90
3040, 2924, 2854, 1723, 1627 cm−1. H NMR
1
1
(12 H, t, J 6.6 Hz), 1.21–1.75 (84 H, m), 2.03
(3 H, s), 2.33–2.40 (2 H, m), 3.20 (3 H, s),
3.22–4.00 (18 H, m), 4.19–4.22 (2 H, m), 4.29
(1 H, m), 4.56 (1 H, q, J 8.8 Hz), 4.75 (1 H, d,
J 8.1 Hz), 7.19–7.40 (10 H, m). FABMS
(positive-ion): m/z 1497 [M+Na]+; 1475
[M+H]+. HRFABMS (positive-ion): m/z
Calcd for C82H143N2NaO18P: 1497.9971;
Found: 1497.9939.
(400 MHz, 5:1 CD3OD–CDCl3) l 0.89 (12 H,
t, J 6.6 Hz), 1.29–1.60 (80 H, m), 1.71–1.73 (4
H, m), 2.00 (3 H, s), 2.32 (1 H, dd, J 8.1, 14.6
Hz), 2.38 (1 H, dd, J 4.4, 14.6 Hz), 3.40–3.49
(5 H, m), 3.52–3.70 (5 H, m), 3.74–3.93 (8 H,
m), 4.03–4.14 (2 H, m), 4.23 (1 H, dd, J 5.1,
9.5 Hz), 4.49 (1 H, d, J 5.1 Hz), 4.59 (1 H, d,
J 8.1 Hz). FABMS (positive-ion): m/z 1331
[M+Na]+; 1309 [M+H]+. HRFABMS
(positive-ion): m/z Calcd for C69H133N2-
NaO18P: 1331.9189; Found: 1331.9203. Anal.
Calcd for C69H133N2O18P (1309.8): C, 63.27;
H, 10.24; N, 2.14; P, 2.37. Found: C, 62.92; H,
10.21; N, 2.04; P, 2.01.
2,6-Anhydro-7-O-[2-acetamido-2,6-dideoxy-
4-O-(diphenylphosphono)-3-O-[(R)-3-(dode-
cyloxy)tetradecyl]-6-fluoro-i-
syl]-3-[(R)-3-(hydroxy)tetradecanamido]-4-O-
[(R) - 3 - (hydroxy)tetradecyl] - 3 - deoxy -
glycero- -ido-heptonic acid (70).—Compound
D
-glucopyrano-
D
-
D
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-3-O-
[(R)-3-(dodecyloxy)tetradecyl]-6-O-methyl-4-
67 (75.2 mg, 0.042 mmol) was treated as de-
scribed in the formation of 68 from 65 to give
70 (48.3 mg, 80%) as an amorphous solid. IR
wmax(KBr) 3323 (broad), 3072, 2924, 2854,
O-phosphono-i-
(hydroxy)tetradecanamido] - 4 - O - [(R) - 3-
(hydroxy)tetradecyl]-3-deoxy- -glycero- -ido-
D
-glucopyranosyl]-3-[(R)-3-
D
D
1729, 1656 cm−1
.
1H NMR (400 MHz,
heptonic acid (72).—Compound 69 (95.4 mg,
0.065 mmol) was treated as described in the
formation of 71 from 68 to give 72 (85.4 mg,
quantitatively) as a white powder, mp 181–
183 °C. [h]2D4 −28.6° (c 0.45, CHCl3). IR
wmax(KBr) 3304 (broad), 3078, 2956, 2923,
CD3OD) l 0.90 (12 H, t, J 6.6 Hz), 1.20–1.76
(84 H, m), 2.00 (3 H, s), 2.30–2.39 (2 H, m),
3.25–3.36 (3 H, m), 3.45 (1 H, t, J 8.1 Hz),
3.60 (1 H, t, J 7.3–9.5 Hz), 3.68–3.94 (10 H,
m), 4.03 (1 H, m), 4.19–4.22 (2 H, m), 4.36–
4.58 (4 H, m, containing 1 H, d, J 5.1 Hz, at
l 4.49), 4.68 (1 H, d, J 8.1 Hz), 7.18–7.41 (10
H, m). FABMS (positive-ion): m/z 1485 [M+
Na]+; 1463 [M+H]+. HRFABMS (positive-
ion): m/z Calcd for C81H140FN2NaO17P:
1485.9771; Found: 1485.9769.
1
2854, 1733, 1654 cm−1. H NMR (400 MHz,
5:1 CD3OD–CDCl3) l 0.89 (12 H, t, J 6.6
Hz), 1.29–1.59 (80 H, m), 1.72–1.75 (4 H, m),
2.00 (3 H, s), 2.32 (1 H, dd, J 8.1, 14.6 Hz),
2.39 (1 H, dd, J 4.4, 14.6 Hz), 3.40 (3 H, s),
3.35–4.04 (19 H, m), 4.09 (1 H, q, J 9.5 Hz),
4.24 (1 H, dd, J 5.1, 9.5 Hz), 4.50 (1 H, d, J
5.1 Hz), 4.59 (1 H, d, J 8.1 Hz). FABMS
(positive-ion): m/z 1345 [M+Na]+; 1323
[M+H]+. HRFABMS (positive-ion): m/z
Calcd for C70H135N2NaO18P: 1345.9345;
Found: 1345.9336. Anal. Calcd for C70H135N2-
O18P (1323.8): C, 63.51; H, 10.28; N, 2.12; P,
2,6-Anhydro-7-O-[2-acetamido-2-deoxy-3-O-
[(R) - 3 - (dodecyloxy)tetradecyl] - 4 - O - phos-
phono - i -
(hydroxy)tetradecanamido] - 4 - O - [(R) - 3-
(hydroxy)tetradecyl]-3-deoxy- -glycero- -ido-
D
- glucopyranosyl] - 3 - [(R) - 3-
D
D
heptonic Acid (71).—A solution of 68 (56.4
mg, 0.039 mmol) in THF (3 mL) containing

230
Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
2.34. Found: C, 63.23; H, 10.14; N, 1.94; P,
2.05.
well plates (Corning, Cambridge, MA), and
were cultured in the presence of TPA (30
ng/mL) for 72 h at 37 °C. After removing the
supernatant, the cells were incubated in 200
mL of fresh RPMI-1640 medium containing
10% NBCS, in the absence or the presence of
30 ng/mL of LPS with graded concentrations
of the compounds in a humidified atmosphere
of 5% CO2 for 4.5 h at 37 °C. After incuba-
tion, the amount of TNFa produced in the
culture supernatants was determined using the
TNFa ELISA kits. As a control, the amount
of TNFa produced by the U937 cells, which
were stimulated with 30 ng/mL of LPS in the
absence of compounds, was used. The concen-
trations (nM) of compounds required to in-
hibit the LPS-induced TNFa production by
U937 cells by 50% (IC50) was calculated from
the control amount. All experiments were car-
ried out at least twice, showing that the data
are reproducible.
2,6-Anhydro-7-O-[2-acetamido-2,6-dideoxy-
3-O-[(R)-3-(dodecyloxy)tetradecyl]-6-fluoro-
4-O-phosphono-i-
(hydroxy)tetradecanamido] - 4 - O - [(R) - 3-
(hydroxy)tetradecyl]-3-deoxy- -glycero- -ido-
D
-glucopyranosyl]-3-[(R)-3-
D
D
heptonic acid (73).—Compound 70 (46.5 mg,
0.032 mmol) was treated as described in the
formation of 71 from 68 to give 73 (39.3 mg,
94%) as a white powder, mp 199.0–200.5 °C.
[h]2D4 −34.5° (c 0.70, CHCl3). IR wmax(KBr)
3399 (broad), 3306, 3085, 2957, 2923, 2854,
1
1733, 1645 cm−1. H NMR (400 MHz, 5:1
CD3OD–CDCl3) l 0.89 (12 H, t, J 6.6 Hz),
1.29–1.60 (80 H, m), 1.69–1.77 (4 H, m), 2.00
(3 H, s), 2.33 (1 H, dd, J 8.1, 14.6 Hz), 2.38 (1
H, dd, J 4.4, 14.6 Hz), 3.40–3.49 (4 H, m),
3.55–4.06 (13 H, m), 4.11 (1 H, q, J 9.5 Hz),
4.23 (1 H, dd, J 5.1, 9.5 Hz), 4.49 (1 H, d, J
5.1 Hz), 4.54–4.80 (3 H, m, containing 1 H, d,
J 8.1 Hz, at l 4.63). FABMS (positive-ion):
m/z 1333 [M+Na]+; 1311 [M+H]+. HR-
FABMS (positive-ion): m/z Calcd for
References
C69H132FN2NaO17P:
1333.9145;
Found:
1333.9158. Anal. Calcd for C69H132FN2O17P
(1311.8): C, 63.18; H, 10.14; F, 1.45; N, 2.14;
P, 2.36. Found: C, 63.31; H, 10.18; F, 1.46; N,
2.06; P, 2.08.
1. Westphal, O.; Luderitz, O. Angew. Chem. 1954, 66, 407–
417.
2. Galanos, C.; Luderitz, O.; Rietschel, E. T.; Westphal, O.
Int. Re6. Biochem. 1977, 14, 239–335.
3. (a) Matsuura, M.; Kojima, Y.; Homma, J. Y.; Kubota,
Y.; Yamamoto, A.; Kiso, M.; Hasegawa, A. FEBS Lett.
1984, 167, 226–230;
(b) Kiso, M.; Ishida, H.; Hasegawa, A. Agric. Biol. Chem.
1984, 48, 251–252;
(c) Kiso, M.; Tanaka, S.; Fujita, M.; Fujishima, Y.;
Ogawa, Y.; Ishida, H.; Hasegawa, H. Carbohydr. Res.
1987, 162, 127–140;
(d) Kiso, M.; Ogawa, Y.; Tanaka, S.; Fujishima, Y.;
Fujita, M.; Tanaka, S.; Hasegawa, A. J. Carbohydr.
Chem. 1987, 6, 625–638.
4. Methods for measurement of biological
activity
The sources of the materials used in the
study are as follows: lipopolysaccharide (LPS)
from E. coli serotype 026:B6 and 12-O-tetra-
decanoylphorbor acetate (TPA) were from
Sigma, St. Louis, MO; RPMI-1640 medium,
fetal bovine serum (FBS), and newborn calf
serum (NBCS) were from Gibco, Grand Is-
land, NY; and human tumor necrosis factor-a
enzyme-linked immunosorbent assay (TNFa
ELISA) kit was from Genzyme, Cambridge,
MA.
4. Zhang, M.; Tracey, K. L. In Endotoxin in Health and
Disease; Brade, H.; Opal, S. M.; Vogel, S. N.; Morrison,
D. C., Eds.; Marcel Dekker: New York, 1999; pp. 915–
926 Chapter 63.
5. Fukushima, D.; Shibayama, S.; Tada, H. Ono Pharm.
Co. Ltd., Patent, PCT International, WO 9965480, Dec.
23, 1999.
6. Rossignol, D. P.; Hawkins, L. D.; Christ, W. J.;
Kobayashi, S.; Kawata, T.; Lynn, M.; Yamatsu, I.;
Kishi, Y. In Endotoxin in Health and Disease; Brade, H;
Opel, S. M.; Vogel, S. N.; Morrison, D. C., Eds.; Marcel
Dekker: New York, 1999; pp. 699–717 Chapter 47.
7. (a) Qureshi, N.; Honovich, J. P.; Hara, H.; Cotter, R. J.;
Takayama, K. J. Biol. Chem. 1988, 263, 5502–5504;
(b) Qureshi, N.; Takayama, K.; Kurtz, R. Infect. Im-
munol. 1991, 59, 441–444;
Cell culture.—Human monoblastic U937
cells were maintained in RPMI-1640 medium
supplemented with 10% FBS, 100 U/mL of
penicillin and 100 mg/mL of streptomycin
(growth medium).
(c) Qureshi, N.; Takayama, K.; Meyer, K. C.; Kirkland,
T. N.; Bush, C. A.; Chen, L.; Wang, R.; Cotter, R. J. J.
Biol. Chem. 1991, 266, 6532–6538;
Production of TNFa by U937 cells: U937
cells (1×104/200 mL/well) were plated in 96-

Y. Watanabe et al. / Carbohydrate Research 333 (2001) 203–231
231
Maeda, H.; Kurakata, S. Carbohydr. Res. 1996, 283, 27–
51.
10. Shiozaki, M.; Kurakata, S.; Tatsuta, T.; Maeda, H.;
Nishijima, M. Tetrahedron 1997, 53, 16041–16060.
11. (a) Mochizuki, T.; Iwano, Y.; Shiozaki, M.; Kurakata, S.;
Kanai, S.; Nishijima, M. Carbohydr. Res. 2000, 324, 225–
230;
(d) Christ, W. J.; McGuinness, P. D.; Asano, O.; Wang,
Y.; Mullarkey, M. A.; Perez, M.; Hawkins, L. D.; Blythe,
T. A.; Dubuc, G. R.; Robidoux, A. L. J. Am. Chem. Soc.
1994, 116, 3637–3638;
(e) Kaltashov, I. A.; Doroshenco, V.; Cotter, R. J.;
Katayama, K.; Qureshi, N. Anal. Chem. 1997, 69, 2317–
2322.
(b) Mochizuki, T.; Iwano, Y.; Shiozaki, M.; Kurakata,
S.; Kanai, S.; Nishijima, M. Tetrahedron. 2000, 56, 7691–
7703.
8. Christ, W. J.; Asano, O.; Robidoux, A. L. C.; Perez, M.;
Wang, Y.; Dubuc, G. R.; Gavin, W. E.; Hawkins, L. D.;
McGuinness, P. D.; Mullarkey, M. A.; Lewis, M. D.;
Kishi, Y.; Kawata, T.; Bristol, J. R.; Rose, J. R.; Rossig-
nol, D. P.; Kobayashi, S.; Hishinuma, I.; Kimura, A.; As-
akawa, N.; Katayama, K.; Yamatsu, I. Science 1995, 268,
80–83.
12. Shiozaki, M.; Kobayashi, Y.; Arai, M.; Watanabe, T.;
Hiraoka, T. J. Med. Chem. 1991, 34, 2643–2646.
13. Watanabe, Y.; Miura, K.; Shiozaki, M.; Kanai, S.;
Kurakata, S.; Nishijima, M. Carbohydr. Res. 2001, 332,
257–277.
14. Shiozaki, M.; Kobayashi, Y.; Ishida, N.; Arai, M.; Hi-
raoka, T.; Nishijima, M.; Kuge, S.; Otsuka, T.; Aka-
matsu, Y. Carbohydr. Res. 1991, 222, 57–68.
9. Shiozaki, M.; Deguchi, N.; Macindoe, W. M.; Arai, M.;
Miyazaki, H.; Mochizuki, T.; Tatsuta, T.; Ogawa, J.;
.