Synthesis of GLA-60 type pyran carboxylic acids with an alkyl chain instead of an ester chain as LPS-antagonists

Article abstract of DOI:10.1016/S0008-6215(01)00055-6

Synthesis of GLA-60 type pyran carboxylic acid analogues with an alkyl chain instead of an ester chain and their LPS-antagonist activity toward human U937 cells are described.

Full text of DOI:10.1016/S0008-6215(01)00055-6

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Carbohydrate Research 332 (2001) 257–277
Synthesis of GLA-60 type pyran carboxylic acids with
an alkyl chain instead of an ester chain as
LPS-antagonists
Yukiko Watanabe,a Kumiko Miura,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 4 December 2000; accepted 9 February 2001
Abstract
Synthesis of GLA-60 type pyran carboxylic acid analogues with an alkyl chain instead of an ester chain and their
LPS-antagonist activity toward human U937 cells are described. © 2001 Elsevier Science Ltd. All rights reserved.
Keywords: Lipid A; Endotoxin; LPS-antagonist; TNFa production inhibitor
is a hydrophobic anchor substance holding an
essentially linear polysaccharide chain to the
cell wall. In recent years, endotoxin-related
compounds have been studied as LPS-antago-
nists that may have potential as immunosup-
pressants,3 or in the treatment of
inflammation, autoimmune diseases3 and
septicemia4, by deactivating LPS-induced ag-
gressive macrophages. For example, Lipid
IVa, which is a biosynthetic precursor of Lipid
A, acts as an LPS-antagonist in the human
body.5 Furthermore, Qureshi’s group6 isolated
a lipid A-related compound from Rhodobacter
sphaeroides that showed potent LPS antago-
nist activity, and recently the Eisai group de-
veloped a related compound, E5564,4,7 as a
highly potent anti-septicemia drug.
1. Introduction
Lipopolysaccharides (LPS)1 cover the outer
surface membrane of gram-negative bacteria
such as Escherichia coli and are highly potent
stimulators of the immune system.2 A variety
of responses, both beneficial and harmful, can
be elicited by LPS. One of these harmful
responses is fatal endotoxic shock (bacterial
sepsis) caused as a consequence of acute in-
flammatory response, which 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, in the terminal disaccharide
phospholipid subunit known as lipid A, which
During our investigation of the biological
activities of compounds related to GLA-60,8
which is a non-reducing monosaccharide ana-
* Corresponding author. Tel.: +81-334-913131; fax: +81-
354-368570.
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)00055-6

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Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
should be more stable than the axial. DIBAL
reduction of compounds 2a and 2b in CH2Cl2
gave 3a and 3b, respectively. Mesylation of 3a
and 3b yielded 4a and 4b, respectively.
Next, in Scheme 2, C-7 hydroxy compounds
17aa, 17ab, 17ac, and 17ba were synthesized
from 5. The starting amine 511 was converted
to an azide 6 by the reported method.12 Alkyl-
ation of the alcohol of 6 with (R)-3-dodecyl-
Fig. 1. Monosaccharide LPS-antagonists toward human U937
cells.
oxy-1-(methanesulfonyloxy)tetradecane
or
(R) - 3 - benzyloxy - 1 - (methanesulfonyloxy)-
tetradecane and sodium hydride gave an ether
7a or 7b, respectively. The anomeric allyl pro-
tecting groups of 7a and 7b were deprotected
to give 8a and 8b, respectively, by treatment
with SeO2 for 8a from 7a, or by treatment
logue of Lipid A, 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 A
and the pyran carboxylic acid B exhibited
fairly strong LPS-antagonistic activity9 toward
human U937 cells. By using this information
on the characteristics of A and B, we synthe-
sized nine compounds in which the ester
bonds were replaced by ether bonds10a in
order to stabilize the compounds and to aim
at increasing their activities. In this paper, we
report the synthesis of these compounds as
LPS-antagonists and their activities (Fig. 1).
with
1,5-cyclooctadienebis(methyldiphenyl-
phosphine)iridium hexafluorophosphate13 as a
catalyst, and successive treatment with water–
pyridine–iodine for 8b from 7b. The reaction
of 8a and 8b with trichloroacetonitrile and a
catalytic amount of 1,8-diazobicyclo[5.4.0]-
undec-7-ene (DBU) gave an anomeric mixture
of trichloroacetimidates 9a and 9b, respec-
tively.14 The anomeric a-cyanation was per-
formed by a previously reported method.15
Treatment of 9a and 9b with trimethylsilyl
2. Results and discussion
,
cyanide using 4 A molecular sieves (4AMS)
and trifluoromethanesulfonate as a catalyst
stereospecifically gave the nitriles 10a and 10b,
respectively. Reduction of the azides of 10a
and 10b with triphenylphosphine and aqueous
ammonia, and successive acylation of the
amines thus obtained with (R)-3-benzyloxy-
tetradecanoic acid, 2,2-difluorotetradecanoic
acid16 or (R)-3-dodecyloxytetradecanoic acid
Synthesis.—First, in Scheme 1, two mesy-
lates 4a and 4b as side chain materials were
prepared from chiral starting diol 1. The start-
ing diol 1, easily obtained by lithium alu-
minum hydride reduction10b in THF of
commercially available (R)-3-hydroxytetra-
decanoic acid, was treated with dodecyl alde-
hyde or benzaldehyde dimethyl acetal using
p-TsOH·water as a catalyst to yield single
acetal 2a or 2b, respectively. The configura-
tions of all substituents of compounds 2a and
2b should be equatorial toward the 1,3-diox-
ane ring, because the equatorial configuration
using
dicyclohexylcarbodiimide
(DCC)-
dimethylaminopyridine (DMAP), or acylation
with tetradecanoyl chloride and triethylamine
gave amides 11aa–11bb. Hydration of the ni-
trile parts of 11aa–11bb with water and 4 M
Scheme 1. Reagents and conditions: (a) n-C11H23CHO or PhCH(OMe)2, p-TsOH, PhCH3, 60 °C, 30 min, 96% (2a) or 97% (2b);
(b) DIBAL, CH2Cl2, rt, 5 h, 89% (3a) and 95% (3b), respectively; (c) MsCl, Et3N, CH2Cl2, 0 °C, 30 min, 98% (4a) and 96% (4b),
respectively.

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
259
Scheme 2. Reagents and conditions: (a) 0.4 M TfN3 in CH2Cl2, DMAP, MeOH, rt, 18 h, 91%; (b) (R)-3dodecyloxy-1-(methane-
sulfonyloxy)tetradecane or (R)-3-benzyloxy-1-(methanesulfonyloxy)tetradecane, NaH, DMF, 73–74%; (c) SeO2, cat. AcOH,
dioxane, reflux, 1 h, 70% (8a from 7a); or [C8H12Ir(PMePh2)2]PF6, THF, rt, 3 h, then water, I2, pyridine, rt, 12 h, 65% (8b from
7b); (d) Cl3CCN, cat. DBU, CH2Cl2, 0 °C, 1 h, 91% (9a from 8a); 0 °C, 2 h, 88% (9b from 8b); (e) TMSCN, cat. TMSOTf, 4AMS,
CH2Cl2, rt, 3 h, 75% (10a from 9a); rt, 2 h, 93% (10b from 9b); (f) (1) Ph3P, THF, 28% NH3 in water, 60 °C, 24 h, (2)
(R)-3-(benzyloxy)tetradecanoic acid, DCC, DMAP, CH2Cl2, rt, 18 h, 79% (11aa from 10a) and 63% (11bb from 10b), or
2,2-difluorotetradecanoic acid, DCC, DMAP, CH2Cl2, rt, 18 h, 94% (11ab from 10a), or tetradecanoyl chloride, Et3N, CH2Cl2,
rt, 1 h, 84% (11ac from 10a), or (R)-3-(dodecyloxy)tetra-decanoic acid, DCC, DMAP, rt, 2 h, 74% (11ba from 10b); (g) 4 M HCl
dioxane, water, 60 °C (80 °C in the case of 11bb), 3–4 h (72 h in the case of 11bb), then Ph2CN2, DMF, 60 °C, 1–2 h, 50% (12aa
from 11aa), 41% (12ab from 11ab), 46% (12ac from 11ac), 26% (12ba from 11ba), 43% (12bb from 11bb); (h) TBDMSCl, DMAP,
CH2Cl2, rt, 3 h, 87% (13aa from 12aa), 88% (13ab from 12ab), 89% (13ac from 12ac), and 62% (13ba from 12ba); (i)
(PhO)2P(O)Cl, DMAP, CH2Cl2, rt, 2 h, 95–97% (\80% in 14ba); (j) 3 M HCl aq, THF, rt, 6 h, 88–93% (15ba, two steps 80%
from 13ba); (k) H2, 20% Pd(OH)2/C in the case of 15aa and 15ba, or 10% Pd/C in the case of 15ab and 15ac, EtOH, rt, 2–16
h, 16aa: 82%, 16ab: 78%, 16ac: 69%, 16ba: 58%; (l) H2, PtO2, THF, rt, 18 h, 17aa: 74%, 17ab: 96%, 17ac: 99%, and 17ba: 86%.

260
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
Scheme 2. (Continued)
hydrogenolysis using Pd(OH)2-on-carbon as a
catalyst, or that of the benzhydryl esters of
18b and 18c using Pd/C as a catalyst produced
carboxylic acids 19a–c. Hydrogenolysis of the
phosphates 19a–c using PtO2 as a catalyst
yielded 5-O-phosphono carboxylic acids 20a–
c, respectively.
Last, in Scheme 4, 7-C-fluoro derivatives
23a and 23b were synthesized from 15aa and
15ab, respectively. Fluorination of the C-7
primary alcohol of 15aa and 15ab with diethyl-
aminosulfur trifluoride (DAST) gave 21a and
21b, respectively. Deprotection of the benzhy-
dryl ester and benzyl ether of 21a by hy-
drogenolysis using Pd(OH)2-on-carbon as a
catalyst, and that of the benzhydryl ester of
21b using Pd/C as a catalyst produced car-
boxylic acids 22a and 22b. Hydrogenolysis of
the phosphates 22a and 22b using PtO2 as a
catalyst yielded 5-O-phosphono carboxylic
acids 23a and 23b, respectively.
Thus, we were able to synthesize a total of
nine compounds, 17aa, 17ab, 17ac, 17ba, 20a,
20b, 20c, 23a and 23b.
Biological acti6ities.—The inhibitory activ-
ity of the nine synthesized compounds, 17aa,
17ab, 17ac, 17ba, 20a, 20b, 20c, 23a and 23b,
on LPS-induced TNFa production was inves-
tigated in vitro using human monoblastic
U937 cells. The IC50 values (nM) of these nine
HCl in dioxane, and successive esterification
of the acid thus obtained with diphenyldi-
azomethane gave esters 12aa–12bb, respec-
tively. Selective protection of the C-7 alcohol
in diols 12aa–12ba with tert-butyldimethyl-
silyl chloride (TBDMSCl) in dichloromethane
using DMAP as a base gave silyl ethers 13aa–
13ba, respectively. Phosphorylation of the C-5
alcohol of 13aa–13ba with diphenyl
chlorophosphate and DMAP yielded phos-
phates 14aa–14ba, respectively. Desilylation
of 14aa–14ba by aqueous 3 M HCl in THF
gave alcohols 15aa–15ba, respectively. Depro-
tection of the benzhydryl esters and benzyl
ethers of both 15aa and 15ba by hydrogenoly-
sis using Pd(OH)2-on-carbon as a catalyst, or
that of the benzhydryl esters of both 15ab and
15ac using Pd/C as a catalyst produced the
carboxylic acids 16aa–16ba. Finally, hy-
drogenolysis of four phosphates 16aa–16ba
using PtO2 as a catalyst yielded 5-O-phos-
phono
respectively.
carboxylic
acids
17aa–17ba,
Thirdly, in Scheme 3, 7-O-methyl deriva-
tives 20a, 20b and 20c were synthesized from
15aa, 15ab and 15ac, respectively. Methyla-
tion of the C-7 primary alcohol of 15aa–15ac
with trimethyloxonium tetrafluoroborate gave
18a–18c, respectively. Deprotection of the
benzhydryl ester and benzyl ether of 18a by

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
261
Scheme 3. Reagents and conditions: (a) Me3OBF4, DTBMP, CH2Cl2, rt, 3 h, 18a: 92%, 18b: 70%, and 18c: 87%; (b) H2, 20%
Pd(OH)2/C in the case of 18a, and 10% Pd/C in the case of 18b and 18c, EtOH, rt, 2–8 h, 19a: 83%, 19b: 69%, and 19c: 62%;
(c) H2, PtO2, THF, rt, 18 h, 20a: 92%, 20b: 98%, and 20c: 92%.
Scheme 4. Reagents and conditions: (a) DAST, DME, CH2Cl2, 0 °C, 2 h, 21a: 52%, and 2b: 77%; (b) H2, 20% Pd(OH)2/C, EtOH,
rt, 8 h, 72% (22a), or 10% Pd/C, EtOH, rt, 2 h, 62% (22b); (f) H2, PtO2, THF, rt, 18 h, 23a: 69%, and 23b: 93%.
and ether compounds in this series may not be
so large.
compounds, 17aa, 17ab, 17ac, 17ba, 20a, 20b,
20c, 23a and 23b, were 32, 3.6, 12, 98, 70, 1.5,
360, 21, and 5.7 nM, respectively. The IC50
values of compounds 17ab and 20b were
stronger than those of compounds A and B (5
and 5.7 nM, respectively).9a,9b The activity of
compound 23b was equal to that of com-
pound B. These three active compounds 17ab,
20b, and 23b coincidentally have the 2,2-
difluorotetradecanoylamino group as an
amide at the C-3 position. The IC50 values of
ether 17ac and its ester ((R)-3-tetra-
decanoyloxytetradecanoyl) derivative9b were
12 and 7.9 nM, respectively. Therefore, the
difference of inhibitory activity between ester
3. Experimental
Melting points were determined on a
Yanagimoto micro melting point apparatus
and are uncorrected. Optical rotations were
obtained by the use of a JASCO P-1030 polar-
1
imeter. H NMR spectra were recorded with a
JEOL JNM-270 or a JEOL-GSX 400 spec-
trometer using Me4Si as the internal standard.
IR absorption spectra were determined with
an IR A-2 spectrophotometer, and mass spec-

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Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
brine, dried over MgSO4, filtered, and concen-
trated in vacuo to give a mixture that was
chromatographed on a silica gel column. Elu-
tion with 3:1 hexane–EtOAc gave (R)-3-dode-
cyloxy-1-hydroxytetradecane (3a) (14.4 g,
89%) as an oil. IR wmax(CHCl3): 3003, 2928,
tra were obtained with a JMS-700 mass spec-
trometer. Elemental analyses were performed
by the Institute of Science and Technology.
Column chromatography was carried out with
Silica Gel 60 (230–400 mesh ASTM, E.
Merck) under a slightly elevated pressure
(1.1–1.5 atm) for easy elution, and the quan-
tity of silica gel used was 50–100 times the
weight charged on the column. Analytical
chromatography was performed on E. Merck
No. 5715 Silica Gel 60-F254 plates. THF was
distilled in the presence of radical anions gen-
erated by sodium-benzophenone ketyl.
Dichloromethane was dried by being passed
through an ICN Alumina B-Super I, and
DMF and pyridine were dried by storage over
1
2855 cm−1. H NMR (400 MHz, CDCl3): l
0.88 (6 H, t, J 6.6–7.3 Hz), 1.26–1.81 (42 H,
m, containing 38 H, broad s, at l 1.26), 2.83
(1 H, brs, OH), 3.40 (1 H, m), 3.46–3.54 (2 H,
m), 3.71–3.83 (2 H, m). FABMS, (positive-
ion): m/z 399 (M+H)+. HRFABMS (posi-
tive-ion): Calcd for C26H55O2: 399.4202;
Found: 399.4221. Anal. Calcd for C26H54O2
(398.7): C, 78.32; H, 13.65. Found: C, 78.48;
H, 13.88.
,
4 A molecular sieves.
(iii) To a solution of the above obtained 3a
(14.2 g, 35.6 mmol) in CH2Cl2 (70 mL) were
added Et3N (7.4 mL, 53.1 mmol) and
methanesulfonyl chloride (4 mL, 51.7 mmol)
at 0 °C. After stirring for 30 min 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. Elu-
tion with 4:1 hexane–EtOAc gave (R)-3-dode-
cyloxy - 1 - (methanesulfonyloxy)tetradecane
(4a) (16.7 g, 98%) as an oil. IR wmax(CHCl3):
(R)-3-Dodecyloxy-1-(methanesulfonyloxy)-
tetradecane (4a).—(i) To a solution of (R)-
1,3-dihydroxytetradecane (1) (9.91 g, 43.0
mmol) in toluene (150 mL) were added dode-
cyl aldehyde (8.73 g, 47.4 mmol) and p-
TsOH·water (1.64 g, 8.62 mmol). After
stirring at 60 °C for 1 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 that was chromatographed on a silica
gel column. Elution with 19:1 hexane–EtOAc
gave (4R)-2,4-diundecyl-1,3-dioxane (2a) (16.3
g, 96%) as an oil. IR wmax(CHCl3): 2927, 2855,
1
2928, 2855, 1359 cm−1. H NMR (400 MHz,
CDCl3): l 0.88 (6 H, t, J 6.6 Hz), 1.26 (38 H,
broad s), 1.40–1.57 (2 H, m), 1.79–1.97 (2 H,
m), 3.00 (3 H, s), 3.32–3.40 (2 H, m), 3.48 (1
H, m), 4.29–4.39 (2 H, m). FABMS, (positive-
ion): m/z 515 (M+K)+ (on addition of KI);
477 (M+H)+. HRFABMS (positive-ion):
Calcd for C27H56O4SK: 515.3536; Found:
515.3561. Anal. Calcd for C27H56O4S (476.8):
C, 68.02; H, 11.84; S, 6.73. Found: C, 68.06;
H, 11.60; S, 6.21.
(R) - 3 - Benzyloxy - 1 - (methanesulfonyloxy)-
tetradecane (4b).—(i) To a solution of (R)-
1,3-dihydroxytetradecane (1) (5.10 g, 22.1
mmol) in DMF (40 mL) were added benzalde-
hyde dimethyl acetal (10 mL, 66.6 mmol) and
p-TsOH·water (845 mg, 4.40 mmol). After
stirring at rt for 1 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 that was chromatographed on a silica
gel column. Elution with 49:1 cyclohexane–
1
1467, 1142, 1120 cm−1. H NMR (400 MHz,
CDCl3): l 0.88 (6 H, t, J 6.6–7.3 Hz), 1.26–
1.43 (38 H, m), 1.54–1.68 (4 H, m), 3.55 (1 H,
m), 3.72 (1 H, dt, J 2.2, 11.7 Hz), 4.09 (1 H,
dd, J 5.1, 11.7 Hz), 4.48 (1 H, t, J 5.1 Hz).
FABMS, (positive-ion): m/z 419 (M+Na)+,
397 (M+H)+; FABMS, (negative-ion): m/z
395 (M−H)−. HRFABMS (positive-ion):
Calcd for C26H53O2: 397.4046; Found:
397.4054. Anal. Calcd for C26H52O2 (396.7):
C, 78.72; H, 13.21. Found: C, 78.53; H, 13.07.
(ii) To a solution of the above obtained 2a
(16.1 g, 40.6 mmol) in CH2Cl2 (50 mL) was
added DIBAL (1.0 M solution in CH2Cl2, 100
mL, 100 mmol) at 0 °C under N2. After stir-
ring for 5 h at rt 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 1 h at rt, the mixture was
extracted with CH2Cl2, washed with water and

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
263
Allyl 2-azido-2-deoxy-4,6-O-isopropylidene-
i- -glucopyranoside (6).—To a solution of 5
EtOAc gave (4R)-2-phenyl-4-undecyl-1,3-
dioxane (2b) (6.85 g, 97%) as an oil. IR
D
1
wmax(CHCl3): 2927, 2856, 1602, 1114 cm−1. H
(6.35 g, 24.5 mmol) in MeOH (120 mL) was
added DMAP (3.07 g, 25.1 mmol) and 0.4 M
TfN3 in CH2Cl2 (120 mL, 48.0 mmol). After
stirring at rt for 18 h, the reaction mixture was
concentrated in vacuo and chromatographed
on a silica gel column. Elution with 3:2 cyclo-
hexane–EtOAc gave 6 (6.33 g, 91%) as a
solid; mp 67 °C. IR wmax(KBr): 3452, 2996,
NMR (270 MHz, CDCl3): l 0.88 (3 H, t, J 6.6
Hz), 1.26–1.53 (20 H, m), 1.61–1.88 (2 H, m),
3.81 (1 H, m), 3.96 (1 H, dt, J 2.6, 11.9 Hz),
4.27 (1 H, dd, J 4.5, 11.2 Hz), 5.51 (1 H, s),
7.29–7.52 (5 H, m). FABMS, (positive-ion):
+
’
m/z 318 (M ). FABMS, (negative-ion): m/z
317 (M−H)−. HRFABMS (positive-ion):
Calcd for C21H34O2: 318.2559; Found:
318.2549. Anal. Calcd for C21H34O2 (318.5):
C, 79.19; H, 10.76. Found: C, 79.14; H, 10.57.
(ii) To a solution of the above obtained 2b
(6.74 g, 21.2 mmol) in CH2Cl2 (40 mL) was
added DIBAL (1.0 M solution in CH2Cl2, 65
mL, 65.0 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 4:1 cyclohexane–
EtOAc gave (R)-3-benzyloxy-1-hydroxytetra-
decane (3b) (6.47 g, 95%) as an oil. IR
2887, 2113 cm−1
.
1H NMR (400 MHz,
CD3OD): l 1.37 (3 H, s), 1.51 (3 H, s),
3.21–3.26 (2 H, m), 3.40 (1 H, t, J 9.5 Hz),
3.53 (1 H, t, J 9.2 Hz), 3.78 (1 H, t, J 10.6
Hz), 3.86 (1 H, dd, J 5.5, 10.6 Hz), 4.09–4.16
(1 H, m), 4.31–4.36 (1 H, m), 4.45 (1 H, d, J
7.3 Hz), 5.17–5.35 (2 H, m), 5.93 (1 H, m).
FABMS, (positive-ion): m/z 286 (M+H)+.
HRFABMS (positive-ion): m/z: Calcd for
C12H20N3O5: 286.1403; Found: 286.1413.
Anal. Calcd for C12H19N3O5 (285.3): C, 50.52;
H, 6.71; N, 14.73. Found: C, 50.28; H, 6.70;
N, 14.80.
Allyl 2-azido-2-deoxy-3-O-[(R)-3-(dodecyl-
oxy)tetradecyl] - 4,6 - O - isopropylidene - i - -
D
glucopyranoside (7a).—To a solution of 6
(5.15 g, 18.1 mmol) in DMF (50 mL) was
gradually added NaH (60% oil dispersion,
1.36 g, 34.0 mmol) at 0 °C with stirring. After
15 min at 0 °C, (R)-3-dodecyloxy-1-(methane-
sulfonyloxy)tetradecane (7.15 g, 15.0 mmol)
was added to the mixture. After stirring for 20
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 concentrated in vacuo
and chromatographed on a silica gel column.
Elution with 4:1 hexane–EtOAc gave 7a (7.31
g, 73%) as a gum. IR wmax(CHCl3): 2928, 2855,
1
wmax(CHCl3): 3626, 3505, 2928, 2856 cm−1. H
NMR (270 MHz, CDCl3): l 0.88 (3 H, t, J 6.6
Hz), 1.27–1.84 (22 H, m), 2.42 (1 H, brs, OH),
3.60–3.85 (3 H, m), 4.48, 4.61 (2 H, ABq, J
11.3 Hz), 7.25–7.35 (5 H, m); FABMS, (posi-
tive-ion): m/z 321 (M+H)+. HRFABMS
(positive-ion): Calcd for C21H37O2: 321.2794;
Found: 321.2789. Anal. Calcd for C21H36O2
(320.5): C, 78.70; H, 11.32. Found: C, 79.77;
H, 11.80.
(iii) The above obtained 3b was treated as
described in Ref. 17 to give (R)-3-benzyloxy-
1-(methanesulfonyloxy)tetradecane (4b) (7.72
g, 96%) as an oil. IR wmax(CHCl3): 2926, 2854
1
2114 cm−1. H NMR (400 MHz, CDCl3): l
0.88 (6 H, t, J 6.6–7.3 Hz), 1.21–1.56 (46 H,
m, containing 3 H, s, at l 1.39, 3 H, s, at l
1.49), 1.69–1.74 (2 H, m), 3.15–3.31 (2 H, m),
3.33 (1 H, t, J 8.1–9.5 Hz), 3.37–3.47 (3 H,
m), 3.60 (1 H, t, J 9.5 Hz), 3.69–3.80 (2 H,
m), 3.84–3.92 (2 H, m), 4.13 (1 H, dd, J 5.9,
13.2 Hz), 4.32 (1 H, d, J 8.1 Hz), 4.36 (1 H,
dd, J 5.1, 13.2 Hz), 5.22–5.36 (2 H, m), 5.92
(1 H, m). FABMS, (positive-ion): m/z 704
(M+K)+, 666 (M+H)+. HRFABMS (posi-
tive-ion): Calcd for C38H72N3O6: 666.5421;
1
cm−1. H NMR (270 MHz, CDCl3): l 0.88 (3
H, t, J 6.6Hz), 1.27 (18 H, brs), 1.47–1.68 (2
H, m), 1.83–2.05 (2 H, m), 2.94 (3 H, s), 3.59
(1 H, m), 4.28–4.42 (2 H, m), 4.45, 4.58 (2 H,
ABq, J 11.3 Hz), 7.25–7.35 (5 H, m).
FABMS, (positive-ion): m/z 421 (M+Na)+,
399 (M+H)+. HRFABMS (positive-ion):
Calcd for C22H38NaO4S: 421.2389; Found:
421.2391.

264
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
J 5.1 Hz, OH), 3.60–3.97 (5 H, m), 4.60 (0.5
H, dt, J 2.2, 5.1–5.9 Hz), 5.24 (0.5 H, t, J 3.7
Hz). FABMS, (positive-ion): m/z 626 (M+
H)+. HRFABMS (positive-ion): Calcd for
C35H68N3O6: 626.5108; Found: 626.5111.
Anal. Calcd for C35H67N3O6 (625.9): C, 67.16;
H, 10.79; N, 6.71. Found: C, 66.85; H, 10.46;
N, 6.80.
Found: 666.5444. Anal. Calcd for C38H71N3O6
(666.0): C, 68.53; H, 10.75; N, 6.31. Found: C,
68.63; H, 10.83; N, 6.53.
Allyl 2-azido-3-O-[(R)-3-(benzyloxy)tetra-
decyl] - 2 - deoxy - 4,6 - O - isopropylidene - i - -
D
glucopyranoside (7b).—To a solution of 6
(4.52 g, 15.8 mmol) in DMF (60 mL) was
gradually added NaH (55% oil dispersion, 963
mg, 22.0 mmol) at 0 °C with stirring. After 15
min at 0 °C, (R)-3-benzyloxy-1-(methanesul-
fonyloxy)tetradecane (5.74 g, 14.4 mmol) was
added to the mixture. After stirring for 20 h at
rt, the mixture was quenched with water, ex-
tracted with EtOAc, washed with water and
brine, dried over MgSO4, and filtered. The
filtrate was concentrated in vacuo and chro-
matographed on a silica gel column. Elution
with 4:1 cyclohexane–EtOAc gave 7b (6.23 g,
74%) as an oil. IR wmax(CHCl3): 2928, 2856,
2-Azido-3-O-[(R)-3-(benzyloxy)tetradecyl]-
2-deoxy-4,6-O-isopropylidene- -glucopyranose
D
(8b).—To a solution of 7b (6.15 g, 10.5 mmol)
in THF (120 mL) was added [C8H12Ir-
(PMePh2)2]PF6 (450 mg). The air in the reac-
tion flask was completely replaced with nitro-
gen and then further replaced with hydrogen
to activate the iridium complex. Immediately
after 1 min, when the red-colored solution of
the iridium complex had become almost color-
less, the hydrogen was completely replaced
with nitrogen. This solution was stirred for 3 h
at rt. After confirming a double bond shift to
an enol ether (as indicated by a slightly higher
Rf value) from the 1-allyloxy group by TLC,
water (45 mL), pyridine (4.5 mL) and I2 (2.3
g) were added to this solution. After 12 h
stirring at rt, the mixture was concentrated in
vacuo, diluted with EtOAc, washed with 10%
Na2S2O3, satd, aq NaHCO3, and brine, dried
over MgSO4, and concentrated to give a mix-
ture that was purified on a silica gel column.
Elution with 3:1 cyclohexane–EtOAc gave an
amorphous mixture 8b (3.71 g, 65%) as a gum.
1
2114 cm−1. H NMR (270 MHz, CDCl3): l
0.88 (3 H, t, J 6.6 Hz), 1.26–1.62 (26 H, m,
containing 3 H, s, at 1.39, 3 H, s, at 1.47),
1.70–1.84 (2 H, m), 3.13–3.23 (2 H, m), 3.33
(1 H, t, J 8.0–9.6 Hz), 3.53–3.63 (2 H, m),
3.72–3.81 (2 H, m), 3.82–3.95 (2 H, m), 4.13
(1 H, dd, J 6.1, 12.7 Hz), 4.31–4.39 (2 H, m,
containing 1 H, d, J 8.0 Hz, at l 4.32), 4.53 (2
H, s), 5.22–5.38 (2 H, m), 5.94 (1 H, m),
7.25–7.45 (5 H, m). FABMS, (positive-ion):
m/z 588 (M+H)+. HRFABMS (positive-
ion): m/z: Calcd for C33H54N3O6: 588.4013;
Found: 588.4023. Anal. Calcd for C33H53N3O6
(587.8): C, 67.43; H, 9.09; N, 7.15. Found: C,
67.33; H, 8.99; N, 7.31.
1
IR wmax(CHCl3): 2928, 2855, 2114 cm−1. H
NMR (270 MHz, CDCl3): l 0.88 (3 H, t, J 6.6
Hz), 1.26–1.60 (26 H, m, containing 1.5 H, s,
at l 1.39, 1.5 H, s, at l 1.40, 3 H, s, at l 1.48),
1.71–1.90 (2 H, m), 2.93 (0.5 H, broad, OH),
3.18–3.27 (1.5 H, m, containing 0.5 H, OH),
3.30–3.99 (8 H, m), 4.46–4.60 (2.5 H, m,
containing 2 H, s, at l 4.52), 5.23 (0.5 H, d, J
3.3Hz), 7.22–7.37 (5 H, m). FABMS, (posi-
tive-ion): m/z 548 (M+H)+. HRFABMS
2-Azido-2-deoxy-3-O-[(R)-3-(dodecyloxy)-
tetradecyl] - 4,6 - O - isopropylidene -
D
- gluco-
pyranose (8a).—A suspension of a mixture of
7a (6.10 g, 9.75 mmol), SeO2 (1.33 g, 12.0
mmol) and AcOH (0.5 mL) in 1,4-dioxane (40
mL) was heated under reflux for 1 h. The
mixture was filtered and insoluble material
was washed with 1,4-dioxane. The filtrate was
concentrated in vacuo and chromatographed
on a silica gel column. Elution with 7:3 hex-
ane–EtOAc gave 8a (4.26 g, 70%) as a gum.
(positive-ion):
Calcd
for
C30H50N3O6:
548.3700; Found: 548.3683. Anal. Calcd for
C33H53N3O6 (547.7): C, 65.79; H, 9.02; N,
7.67. Found: C, 65.43; H, 9.05; N, 7.68.
IR wmax(CHCl3): 3599, 2928, 2855, 2114 cm−1
.
1H NMR (400 MHz, CDCl3): l 0.88 (6 H, t, J
6.6 Hz), 1.21–1.56 (46 H, m, containing 3 H,
s, at l 1.40, 3 H, s, at l 1.49), 1.68–1.81 (2 H,
m), 2.99 (0.5 H, d, J 2.9 Hz, OH), 3.21–3.30
(2 H, m), 3.36–3.48 (4 H, m), 3.50 (0.5 H, d,
2,2,2-Trichloroethylimidoyl
oxy-3-O-[(R)-3-(dodecyloxy)tetradecyl]-4,6-
O-isopropylidene-h-
2-azido-2-de-
D
-glucopyranoside (9a).—
To a solution of 8a (4.25 g, 6.79 mmol) and
CCl3CN (7.0 mL, 69.8 mmol) in CH2Cl2 (15

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
265
with 4:1 hexane–EtOAc gave 10a (2.95g, 75%)
as a gum. IR wmax(CHCl3): 2928, 2855, 2119
mL) was added DBU (211 mg, 1.38 mmol) at
0 °C. After stirring for 1 h at 0 °C, satd
NaHCO3 was added to the reaction mixture,
and it was diluted with EtOAc, washed with
water and brine, dried over MgSO4, and
filtered. The filtrate was concentrated in vacuo
and chromatographed on a silica gel column.
Elution with 7:3 hexane–EtOAc gave 9a (4.77
g, 91%) as a gum. IR wmax(CHCl3): 3344, 2928,
1
cm−1. H NMR (400 MHz, CDCl3): l 0.88 (6
H, t, J 6.6–7.3 Hz), 1.21–1.59 (46 H, m,
containing 3 H, s, at l 1.42, 3 H, s, at l 1.49),
1.71–1.79 (2 H, m), 3.34–3.46 (3 H, m), 3.59–
3.82 (6 H, m), 3.91–4.00 (2 H, m), 4.78 (1 H,
d, J 5.9 Hz). FABMS, (positive-ion): 635
(M+H)+. HRFABMS (positive-ion): Calcd
for C36H67N4O5: 635.5133; Found: 635.5111.
Anal. Calcd for C36H66N4O5 (634.9): C, 68.10;
H, 10.48; N, 8.82. Found: C, 67.67; H, 10.46;
N, 8.93.
1
2855, 2115, 1674 cm−1. H NMR (400 MHz,
CDCl3): l 0.88 (6 H, t, J 6.6–7.3 Hz), 1.26–
1.58 (46 H, m, containing 3 H, s, at l 1.43, 3
H, s, at l 1.52), 1.71–1.82 (2 H, m), 3.21–3.30
(2 H, m), 3.36–3.46 (3 H, m), 3.58 (dd, 1 H, J
3.7, 9.5 Hz), 3.69–4.01 (7 H, m), 6.31 (1 H, d,
J 3.7 Hz), 8.70 (1 H, s). FABMS, (positive-
ion): m/z 791 (M+Na)+. HRFABMS (posi-
tive-ion): Calcd for C37H67Cl3N4NaO6:
791.4024; Found: 791.4002.
2,6-Anhydro-3-azido-4-O-[(R)-(benzyloxy)-
tetradecyl]-3-deoxy-5,7-O-isopropylidene-
glycero- -ido-heptononitrile
D
-
D
(10b).—Com-
pound 9b (4.05 g, 5.85 mmol) was treated as
described above to give 10b (3.04 g, 93%) as a
gum. IR wmax(CHCl3): 2928, 2856, 2119 cm−1
.
2,2,2-Trichloroethylimidoyl
[(R) - 3 - (benzyloxy)tetradecyl] - 2 - deoxy - 4,6-
O-isopropylidene-h- -glucopyranoside (9b).—
2-azido-3-O-
1H NMR (270 MHz, CDCl3): l 0.88 (3 H, t, J
6.6 Hz), 1.26–1.58 (26 H, m, containing 3 H,
s, at l 1.41, 3 H, s, at l 1.47), 1.70–1.91 (2 H,
m), 3.52–4.05 (9 H, m), 4.54, 4.48 (2 H, ABq,
J 11.6 Hz), 4.78 (1 H, d, J 5.4 Hz), 7.24–7.36
(5 H, m). FABMS, (positive-ion): m/z 579
(M+Na)+, 557 (M+H)+. HRFABMS (pos-
itive-ion): Calcd for C31H48NaN4O5: 579.3522;
Found: 579.3505. Anal. Calcd for C31H48N4O5
(556.7): C, 66.88; H, 8.69; N, 10.06. Found: C,
66.79; H, 8.91; N, 10.00.
D
Compound 8b (3.66 g, 6.70 mmol) was treated
as described above to give 9b (4.08 g, 88%) as
a gum. IR wmax(CHCl3): 2116, 1676, 1603
1
cm−1. H NMR (270 MHz, CDCl3): l 0.88 (3
H, t, J 6.6 Hz), 1.26–1.65 (26 H, m, contain-
ing 3 H, s, at l 1.41, 3 H, s, at l 1.50),
1.73–1.95 (2 H, m), 3.52–4.07 (9 H, m), 4,53
(2 H, s), 6.31 (1 H, d, J 3.8 Hz), 7.25–7.35 (5
H, m), 8.70 (1 H, s, NH); FABMS, (positive-
ion): m/z 713 (M+Na)+(on addition of NaI).
HRFABMS (positive-ion): Calcd for C32H49-
N4O6Cl3Na: 713.2615; Found: 713.2606. Anal.
Calcd for C32H49Cl3N4O6 (692.1): C, 55.53; H,
7.14; Cl, 15.37; N, 8.10. Found: C, 55.53; H,
6.78; Cl, 15.45; N, 7.93.
2,6-Anhydro-3-[(R)-3-(benzyloxy)tetrade-
canamido]-3-deoxy-4-O-[(R)-3-(dodecyloxy)-
tetradecyl]-5,7-O-isopropylidene-
D
-glycero- -
D
ido-heptononitrile (11aa).—To a solution of
10a (1.01 g, 1.59 mmol) in THF (8 mL) was
added PPh3 (438 mg, 1.67 mmol). After stir-
ring for 1 h at rt, 28% NH3 in water (4 mL)
was added to this solution. The mixture was
stirred for 24 h at 60 °C, concentrated in
vacuo, diluted with toluene, and concentrated
in vacuo again. This procedure was repeated
three times to completely remove the water.
The residual mixture was dissolved in CH2Cl2
(10 mL). To this solution, (R)-3-(benzyl-
oxy)tetradecanoic acid (640 mg, 1.91 mmol),
DCC (496 mg, 2.40 mmol), and DMAP (297
mg, 2.43 mmol) were added with stirring.
After 18 h at rt, the reaction mixture was
filtered, and the filtrate was concentrated in
vacuo to give a mixture. The mixture was
chromatographed on a silica gel column. Elu-
2,6-Anhydro-3-azido-3-deoxy-4-O-[(R)-3-
(dodecyloxy)tetradecyl]-5,7-O-isopropylidene-
D
-glycero- -ido-heptononitrile (10a).—To a
D
solution of 9a (4.75 g, 6.17 mmol) and
Me3SiCN (1.30 mL, 9.75 mmol) in CH2Cl2 (15
mL) was added 4AMS (3.55 g). After stirring
for 1 h at rt, TMSOTf (0.35 mL, 1.93 mmol)
was added to the suspension. After stirring for
3 h at rt, satd NaHCO3 was added. The
reaction mixture was filtered through celite,
diluted with EtOAc, washed with water and
brine, dried over MgSO4, and filtered. The
filtrate was concentrated in vacuo and chro-
matographed on a silica gel column. Elution

266
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
tetradecanamido-
D
-glycero- -ido-heptononi-
D
tion with 4:1 hexane–EtOAc gave 11aa (1.16
g, 79%) as a gum. IR wmax(CHCl3): 3328, 2928,
trile (11ac).—Compound 10a (926 mg, 1.46
mmol) was treated as described above to give
a mixture of a product amine and spent
reagents, which was dissolved in CH2Cl2 (5
mL). To this solution, Et3N (0.3 mL, 2.13
mmol) and tetradecanoyl chloride (0.6 mL,
2.13 mmol) were added at 0 °C. After stirring
for 1 h at rt, the reaction mixture was diluted
with EtOAc, washed with water, satd
NaHCO3, and brine, dried over MgSO4, and
filtered. The filtrate was concentrated in vacuo
and chromatographed on a silica gel column.
Elution with 4:1 hexane–EtOAc gave 11ac
(999 mg, 84%) as a gum. IR wmax(CHCl3):
2855, 1672 cm−1
.
1H NMR (400 MHz,
CDCl3): l 0.88 (9 H, t, J 5.9–7.3 Hz), 1.11–
1.70 (68 H, m, containing 3 H, s, at l 1.40, 3
H, s, at l 1.47) 2.41 (1 H, dd, J 5.9, 15.4 Hz),
2.60 (1 H, dd, J 3.7, 15.4 Hz), 3.18–3.33 (5 H,
m), 3.58–3.80 (5 H, m), 3.88–4.00 (2 H, m),
4.56, 4.59 (2 H, ABq, J 11.0 Hz), 5.27 (1 H, d,
J 5.9 Hz), 7.20 (1 H, d, J 5.1 Hz, NH),
7.26–7.39 (5 H, m). FABMS, (positive-ion):
m/z 925 (M+H)+. HRFABMS (positive-
ion): Calcd for C57H101N2O7: 925.7609;
Found: 925.7619. Anal. Calcd for C57H100-
N2O7 (925.4): C, 73.98; H, 10.89; N, 3.03.
Found: C, 74.09; H, 10.70; N, 3.12.
3691, 2927, 2855, 1676 cm−1. H NMR (400
1
MHz, CDCl3): l 0.88 (9 H, t, J 6.6–7.3 Hz),
1.21–1.72 (70 H, m containing 3 H, s, at l
1.42, 3 H, s, at l 1.50), 2.23 (2 H, dt, J 3.7, 7.3
Hz), 3.35 (2 H, t, J 6.6–7.3 Hz), 3.46–3.76 (6
H, m), 3.91–4.04 (3 H, m), 5.40 (1 H, d, J 5.9
Hz), 6.49 (1 H, d, J 4.4 Hz, NH). FABMS,
(positive-ion): m/z 819 (M+H)+. HR-
FABMS (positive-ion): Calcd for C50H95N2O6:
819.7190; Found: 819.7203. Anal. Calcd for
C50H94N2O6 (819.3): C, 73.30; H, 11.57; N,
3.42; Found: C, 73.49; H, 11.10; N, 3.49.
2,6-Anhydro-4-O-[(R)-3-(benzyloxy)tetra-
decyl] - 3 - deoxy - 3 - [(R) - 3 - (dodecyloxy)tetra-
2,6-Anhydro-3-deoxy-3-(2,2-difluorotetra-
decanamido) - 4 - O - [(R) - 3 - (dodecyloxy)tetra-
decyl]-5,7-O-isopropylidene-
D
-glycero- -ido-
D
heptononitrile (11ab).—To a solution of 10a
(1.06 g, 1.67 mmol)) in THF (5 mL) was
added PPh3 (451 mg, 1.72 mmol). After stir-
ring for 1 h at rt, 28% NH3 in water (2.5 mL)
was added to this solution. The mixture was
stirred for 20 h at 60 °C, concentrated in
vacuo, diluted with toluene, and concentrated
in vacuo again. This procedure was repeated
three times to remove the water completely.
The residual mixture was dissolved in CH2Cl2
(8 mL). To this solution, 2,2-difluorotetra-
decanoic acid (663 mg, 2.51 mmol), DCC (517
mg, 2.51 mmol), and DMAP (307 mg, 2.51
mmol) were added. After stirring for 18 h at
rt, the reaction mixture was filtered. The
filtrate was concentrated in vacuo and chro-
matographed on a silica gel column. Elution
with 4:1 hexane–EtOAc gave 11ab (1.34 g,
94%) as a gum. IR wmax(CHCl3): 3436, 2928,
decanamido]-5,7-O-isopropylidene-
D
-glycero-
D
-ido-heptononitrile (11ba).—Compound 10b
(1.49 g, 2.67 mmol) was treated as described
above to give a mixture of a product amine
and spent reagents, which was dissolved in
CH2Cl2 (10 mL). To this solution, (R)-3-
dodecyloxytetradecanoic acid (1.32 g, 3.20
mmol), and DCC (0.827 g, 4.01 mmol),
DMAP (0.489 g, 4.01 mmol) was added. After
stirring for 2 h at rt, the mixture was filtered
and concentrated in vacuo to give a mixture.
The mixture was chromatographed on a silica
gel column. Elution with 7:3 hexane–EtOAc
gave 11ba (1.82 g, 74%) as a gum. IR
1
2856, 1713, 1671 cm−1. H NMR (400 MHz,
CDCl3): l 0.88 (9 H, t, J 6.6–7.3 Hz), 1.11–
1.86 (68 H, m, containing 3 H, s, at l 1.43, 3
H, s, at l 1.51), 2.03–2.15 (2 H, m), 3.31–3.43
(3 H, m), 3.56–3.78 (5 H, m), 3.91–4.00 (2 H,
m), 4.05 (1 H, m), 5.27 (1 H, d, J 5.9 Hz), 6.82
(1 H, d, J 5.1 Hz, NH). FABMS, (positive-
ion): m/z 855 (M+H)+. HRFABMS (posi-
tive-ion): Calcd for C50H93F2N2O6: 855.7002;
Found: 855.7004.
wmax(CHCl3): 3304, 2924, 2854, 1657 cm−1. H
1
NMR (400 MHz, CDCl3): l 0.88 (t, 9 H, J
5.9–7.3 Hz), 1.17–1.85 (m, 60 H), 2.26–2.32
(m, 1 H), 2.52–2.48 (m, 1 H), 3.40–3.73 (m, 9
H), 3.90–3.95 (m, 2 H), 4.05–4.09 (m, 1 H),
4.49 (s, 2 H), 5.20 (d, 1 H, J 6.6 Hz), 7.17 (d,
1 H, J 5.8 Hz), 7.26–7.36 (m, 5 H). HR-
FABMS (positive-ion): Calcd for C57H101N2O7
(M+H)+: 925.7609; Found: 925.7594.
2,6-Anhydro-3-deoxy-4-O-[(R)-3-(dode-
cyloxy)tetradecyl] - 5,7 - O - isopropylidene - 3-

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
267
2,6 - Anhydro - 3 - [(R) - 3 - (benzyloxy)tetra-
decanamido] - 4 - O - [(R) - 3 - (benzyloxy)tetra-
Diphenylmethyl 2,6-anhydro-3-[(R)-3-(ben-
zyloxy)tetradecanamido] - 3 - deoxy - 4 - O - [(R)-
decyl]-3-deoxy-5,7-O-isopropylidene-
D
-glycero-
3 - (dodecyloxy)tetradecyl] -
D
- glycero -
D
- ido-
D
-ido-heptononitrile (11bb).—(i) Compound
heptonate (12aa).—A solution of 11aa (1.11 g,
1.20 mmol) in 4 M HCl in dioxane (5 mL) and
water (1 mL) was stirred at 60 °C for 3 h. The
reaction mixture was concentrated and dried
in vacuo to give a mixture that was dissolved
in DMF (6 mL). Ph2CN2 (585 mg, 3.01 mmol)
was added to this solution. This solution was
warmed at 60 °C for 1 h. The mixture was
diluted with EtOAc, washed with satd
NaHCO3 and brine, dried over MgSO4, and
filtered. The filtrate was concentrated in vacuo
and chromatographed on a silica gel column.
Elution with 1:1 hexane–EtOAc gave 12aa
(642 mg, 50%) as a solid; mp 105.5–106.5 °C.
IR wmax(KBr): 3329, 2921, 2851, 1735, 1645
10b (3.02 g, 5.43 mmol) was treated as de-
scribed above to give a mixture, which was
chromatographed on a silica gel column. Elu-
tion with cyclohexane–EtOAc (9:1, then 3:2)
gave an amine (R2=H in 11b, 2.03 g, 71%) as
a gum. IR wmax(CHCl3): 3691, 2928, 2856,
1
1602 cm−1. H NMR (400 MHz, CDCl3): l
0.88 (3 H, t, J 6.9 Hz), 1.26–1.65 (28 H, m,
containing 3 H, s, at l 1.40, 3 H, s, at l 1.46),
2.95 (1 H, dd, J 6.0, 9.7 Hz), 3.22 (1 H, t, J 9.2
Hz), 3.50–3.56 (2 H, m), 3.65–3.76 (3 H, m),
3.91 (1 H, m), 3.99 (1 H, m), 4.46, 4.55 (2 H,
ABq, J 11.7 Hz), 4.74 (1 H, d, J 6.0 Hz),
7.26–7.36 (5 H, m). FABMS, (positive-ion):
m/z 553 (M+Na)+, 531 (M+H)+. HR-
FABMS (positive-ion): Calcd for C31H50-
NaN2O5: 553.3617; Found: 553.3603. Anal.
Calcd for C31H50N2O5 (530.7): C, 70.15; H,
9.50; N, 5.28. Found: C, 69.72; H, 9.47; N,
4.52.
1
cm−1. H NMR (400 MHz, CDCl3): l 0.88 (9
H, t, J 5.9–7.3 Hz), 1.11–1.65 (62 H, m) 2.08
(1 H, t, J 5.9–6.6 Hz, OH), 2.32 (2 H, d, J 5.1
Hz), 3.31–3.43 (5 H, m), 3.49–3.83 (8 H, m,
containing OH), 4.33, 4.38 (2 H, ABq, J 11.3
Hz), 4.65 (1 H, d, J 5.1 Hz), 6.85 (1 H, s), 6.89
(1 H, d, J 8.8 Hz, NH), 7.22–7.35 (15 H, m).
FABMS, (positive-ion): m/z 1092 (M+Na)+,
1070 (M+H)+. HRFABMS (positive-ion):
Calcd for C67H107NaNO9: 1092.7844; Found:
1092.7838. Anal. Calcd for C67H107NO9
(1070.6): C, 75.17; H, 10.07; N, 1.31. Found:
C, 74.95; H, 10.11; N, 1.36
(ii) To a solution of the above obtained
amine (R2 =H in 11b) (1.96 g, 3.71 mmol) in
CH2Cl2 (30 mL) were added (R)-3-(benzyl-
oxy)tetradecanoic acid (1.50 g, 4.48 mmol),
DCC (963 mg, 4.67 mmol) and DMAP (574
mg, 4.70 mmol). After stirring for 18 h at rt,
the mixture was filtered, diluted with EtOAc,
washed with satd NaHCO3 and brine, dried
over MgSO4, filtered, and concentrated in
vacuo to give a mixture. The mixture was
chromatographed on a silica gel column. Elu-
tion with 4:1 cyclohexane–EtOAc gave 11bb
(2.79 g, 89%) as a gum. IR wmax(CHCl3): 3691,
Diphenylmethyl
(2,2 - difluorotetradecanamido) - 4 - O - [(R) - 3-
(dodecyloxy)tetradecyl] - - glycero - - ido-
2,6-anhydro-3-deoxy-3-
D
D
heptonate (12ab).—A solution of 11ab (1.05 g,
1.23 mmol)) in 4 M HCl in dioxane (5 mL)
and concd HCl (1 mL) was stirred at 60 °C for
4 h. The reaction mixture was concentrated
and dried in vacuo to give a mixture that was
dissolved in DMF (5 mL). Ph2CN2 (599 mg,
3.08 mmol) was added to this solution, which
was warmed at 60 °C for 2 h. The mixture was
diluted with EtOAc, washed with satd
NaHCO3 and brine, dried over MgSO4, and
filtered. The filtrate was concentrated in vacuo
and chromatographed on a silica gel column.
Elution with 3:2 hexane–EtOAc gave 12ab
(501mg, 41%) as an amorphous solid. IR
wmax(KBr): 3394, 3329, 2923, 2853, 1736, 1679
1
2928, 2855, 1672, 1603 cm−1. H NMR (270
MHz, CDCl3): l 0.88 (3 H, t, J 6.6 Hz),
1.25–1.77 (48 H, m, containing 3 H, s, at l
1.39, 3 H, s, at l 1.44), 2.35 (1 H, dd, J 6.5,
15.5 Hz), 2.54 (1 H, dd, J 3.6, 15.5 Hz),
3.21–3.30 (2 H, m), 3.40 (1 H, m), 3.56–3.81
(5 H, m), 3.86–4.03 (2 H, m), 4.40, 4.45 (2 H,
ABq, J 11.7 Hz), 4.55 (2 H, s), 5.25 (1 H, d, J
6.0 Hz), 7.10 (1 H, d, J 5.5 Hz, NH), 7.25–
7.40 (10 H, m). FABMS, (positive-ion): m/z
869 (M+Na)+, 847 (M+H)+. HRFABMS
(positive-ion):
Calcd
for
C52H83N2O7:
847.6200; Found: 847.6199. Anal. Calcd for
C52H82N2O7 (847.2): C, 73.72; H, 9.76; N,
3.31. Found: C, 73.50; H, 9.82; N, 3.55.
1
cm−1. H NMR (400 MHz, CDCl3): l 0.88 (9
H, t, J 6.6–7.3 Hz), 1.21–1.55 (60 H, m),

268
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
C67H107KNO9 (M+K)+: 1108.7583; Found:
1108.7576.
1.67–1.71 (2 H, m), 1.99–2.03 (3 H, m, con-
taining OH), 3.32–3.40 (4 H, m), 3.48 (1 H, t,
J 8.8–10.3 Hz), 3.57–3.62 (2 H, m), 3.72–3.79
(3 H, m, containing OH), 3.83 (1 H, m,
changed to dd, J 3.7, 11.7 Hz, on addition of
D2O), 4.39 (1 H, m), 4.58 (1 H, d, J 5.9 Hz),
6.94 (1 H, s), 7.15 (1 H, d, J 9.5 Hz, NH),
7.29–7.41 (10 H, m). FABMS, (positive-ion):
m/z 1038 (M+K)+, 1022 (M+Na)+. HR-
FABMS (positive-ion): Calcd for C60H99F2-
NNaO8: 1022.7236; Found: 1022.7223. Anal.
Calcd for C60H99F2NO8 (1000.4): C, 72.03; H,
9.98; F, 3.80; N, 1.40. Found: C, 71.64; H,
9.82; F, 3.82; N, 1.58.
Diphenylmethyl 2,6-anhydro-3-[(R)-3-(ben-
zyloxy)tetradecanamido]-4-O-[(R)-3-(benzyl-
oxy)tetradecyl] - 3 - deoxy -
D
- glycero -
D
- ido-
heptonate (12bb).—Compound 11bb (2.70 g,
3.19 mmol) was treated, as described in the
formation of 12aa from 11aa to give 12bb
(1.35 g, 43%) as an amorphous solid. IR
wmax(CHCl3): 3691, 2928, 2855, 1672, 1603
1
cm−1. H NMR (270 MHz, CDCl3): l 0.88 (3
H, t, J 6.5 Hz), 1.25–1.71 (42 H, m), 1.95 (1
H, broad, OH), 2.29 (2 H, d, J 5.3 Hz), 3.30 (1
H, broad, OH), 3.35–3.74 (10 H, m), 4.32,
4.38 (2 H, ABq, J 11.3 Hz), 4.43 (2 H, s), 4.62
(1 H, d, J 5.6 Hz), 6.84 (1 H, s), 6.87 (1 H, d,
J 8.8 Hz, NH), 7.23–7.33 (10 H, m). FABMS,
(positive-ion): m/z 1014 (M+Na)+, 992
(M+H)+. HRFABMS (positive-ion): Calcd
for C62H90NO9: 992.6616; Found: 992.6610.
Anal. Calcd for C62H89NO9 (992.4): C, 75.04;
H, 9.04; N, 1.41. Found: C, 74.62; H, 8.94; N,
1.43.
Diphenylmethyl 2,6-anhydro-3-deoxy-4-O-
[(R) - 3 - (dodecyloxy)tetradecyl] - 3 - tetradecan-
amido-
D
-glycero-
D
-ido-heptonate
(12ac).—
Compound 11ac (901 mg, 1.10 mmol) was
treated, as described in the formation of 12aa
from 11aa to give 12ac (485 mg, 46%) as a
white solid; mp 128.5–129.5 °C. IR wmax(KBr):
1
3406, 3332, 1738, 1646 cm−1. H NMR (400
MHz, CDCl3): l 0.88 (6 H, t, J 6.6–7.3 Hz),
1.21–1.73 (64 H, m), 2.07 (2 H, t, J 7.3–8.1
Hz), 3.35 (2 H, t, J 6.6 Hz), 3.39–3.44 (2 H,
m), 3.52–3.63 (4 H, m, containing OH), 3.76–
3.84 (4 H, m, containing OH), 4.43 (1 H, dt, J
5.9, 9.5 Hz), 4.62 (1 H, d, J 5.9 Hz), 6.09 (1 H,
d, J 9.5 Hz, NH), 6.90 (1 H, s), 7.29–7.38 (10
H, m). FABMS, (positive-ion): m/z 986 (M+
Na)+, 964 (M+H)+. HRFABMS (positive-
ion): Calcd for C60H101NNaO8: 986.7425;
Found: 986.7424. Anal. Calcd for C60H101NO8
(964.5): C, 74.72; H, 10.56; N, 1.45. Found: C,
74.41; H, 10.27; N, 1.51.
Diphenylmethyl 2,6-anhydro-3-[(R)-3-(ben-
zyloxy)tetradecanamido] - 3 - deoxy - 7 - O - tert-
butyldimethylsilyl - 4 - O - [(R) - 3 - (dodecyloxy)-
tetradecyl]-
D
-glycero- -ido-heptonate (13aa).
D
—To a solution of 12aa (514 mg, 0.480 mmol)
in CH2Cl2 (3 mL) were added DMAP (88.3
t
mg, 0.723 mmol) and BuMe2SiCl (109 mg,
0.720 mmol). After stirring for 3 h at rt, the
mixture was diluted with EtOAc, washed with
water, satd NaHCO3, and brine, dried over
MgSO4, filtered, and chromatographed on a
silica gel column. Elution with 7:3 hexane–
EtOAc gave 13aa (496 mg, 87%) as a gum. IR
wmax(CHCl3): 3432, 3356, 2928, 2856, 1732,
Diphenylmethyl
2,6-anhydro-4-O-[(R)-3-
(benzyloxy)tetradecyl] - 3 - deoxy - 3 - [(R) - 3-
(dodecyloxy)tetradecanamido] - 5,7 - O - isopro-
1
1667 cm−1. H NMR (400 MHz, CDCl3): l
0.059 (3 H, s), 0.069 (3 H, s), 0.85–0.95 (18 H,
m), 1.26–1.62 (62 H, m), 2.30 (2 H, d, J 5.9
Hz), 3.29–3.43 (5 H, m), 3.50–3.64 (5 H, m,
containing OH), 3.75–3.84 (2 H, m), 4.30–
4.38 (3 H, m, containing 2 H, ABq, J 11.0 Hz,
at l 4.33, 4.37), 4.64 (1 H, d, J 5.1 Hz), 6.82
(1 H, d, J 8.8 Hz, NH), 6.84 (1 H, s), 7.22–
7.35 (15 H, m). FABMS, (positive-ion): m/z
1184 (M+H)+. HRFABMS (positive-ion):
Calcd for C73H122NO9Si: 1184.8889; Found:
1184.8903. Anal. Calcd for C73H121NO9Si
(1184.8): C, 74.00; H, 10.29; N, 1.18. Found:
C, 74.23; H, 10.32; N, 1.18.
pylidene- -ido-heptonate (12ba).—Compound
D
11ba (146 mg, 0.158 mmol) was treated, as
described in the formation of 12aa from 11aa
to give 12ba (44.5 mg, 26%) as a gum. IR
1
wmax(CHCl3): 2927, 2855, 1734, 1669 cm−1. H
NMR (400 MHz, CDCl3): l 0.88 (9 H, t, J
6.6–7.3 Hz), 1.16–1.78 (62 H, m), 2.07–2.35
(2 H, m), 3.18–3.31 (2 H, m), 3.34–3.87 (9 H,
m), 4.34–4.49 (3 H, m), 4.63 (1 H, d, J 5.9
Hz), 6.88 (1 H, s), 6.96 (1 H, d, J 8.8 Hz),
7.24–7.34 (15 H, m). FABMS, (positive-ion):
m/z 1108 (M+K)+ (on addition of KI).
HRFABMS
(positive-ion):
Calcd
for

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
269
tetradecanomido]-
D
-glycero-
D
-ido-heptonate
Diphenylmethyl
2,6-anhydro-7-O-tert-
(13ba).—Compound 12ba (38.8 mg, 0.036
mmol) was treated, as described in the forma-
tion of 13aa from 12aa to give 13ba (26.8 mg,
62%) as a gum. IR wmax(CHCl3): 2919, 2850,
butyldimethylsilyl-3-deoxy-3-(2,2-difluorotetra-
decanamido) - 4 - O - [(R) - 3 - (dodecyloxy)tetra-
decyl]-
D
-glycero-
D
-ido-heptonate
(13ab).—
Compound 12ab (431 mg, 0.430 mmol) was
treated as described in the formation of 13aa
from 12aa to give 13ab (421 mg, 88%) as a
gum. IR wmax(CHCl3): 3692, 3607, 3424, 2928,
1734, 1650 cm−1
.
1H NMR (500 MHz,
CDCl3): l 0.05 (s, 3 H), 0.063 (s, 3 H) 0.82–
0.94 (m, 18 H), 1.26–1.83 (m, 62 H), 2.21–
2.23 (m, 2 H), 3.17–3.29 (m, 2 H), 3.35–3.42
(m, 2 H), 3.50–3.55 (m, 1 H), 3.60–3.65 (m, 2
H), 3.72–3.80 (m, 4 H), 4.21–4.26 (m, 1 H),
4.33–4.37 (m, 1 H), 4.45–4.50 (m, 2 H), 4.63
(d, 1 H, J 5.5 Hz), 6.87–6.89 (m, 2 H),
7.23–7.39 (m, 15 H). FABMS, (positive-ion):
m/z 1222 (M+K)+ (on addition of KI). HR-
FABMS (positive-ion): Calcd for C73H121-
KNO9Si (M+K)+: 1222.8448; Found:
1222.8459.
2856, 1708 cm−1
.
1H NMR (400 MHz,
CDCl3): l 0.07 (3 H, s), 0.08 (3 H, s), 0.84–
0.89 (18 H, m), 1.26–1.50 (60 H, m), 1.67–
1.71 (2 H, m), 1.98–2.06 (2 H, m), 3.31–3.37
(4 H, m), 3.46–3.50 (2 H, m, containing OH,
changed to t, 1 H, J 8.1–10.3 Hz, at l 3.48,
on addition of D2O), 3.65–3.71 (3 H, m), 3.78
(1 H, dd, J 4.4, 11.0 Hz), 3.83 (1 H, dd, J 4.4,
11.0 Hz), 4.37 (1 H, m), 4.57 (1 H, d, J 5.1
Hz), 6.94 (1 H, s), 7.11 (1 H, d, J 8.8 Hz,
NH), 7.28–7.39 (10 H, m). FABMS, (positive-
ion): m/z 1136 (M+Na)+, 1114 (M+H)+.
HRFABMS (positive-ion): Calcd for C66H113-
F2NNaO8Si: 1136.8101; Found: 1136.8101.
Anal. Calcd for C66H113F2NO8Si (1114.7): C,
71.12; H, 10.22; F, 3.41; N, 1.26. Found: C,
70.86; H, 10.02; F, 3.45; N, 1.35.
Diphenylmethyl 2,6-anhydro-3-[(R)-3-(ben-
zyloxy)tetradecanamido] - 3 - deoxy - 7 - O - tert-
butyldimethylsilyl-5-O-diphenylphosphono-4-O-
[(R) - 3 - (dodecyloxy)tetradecyl] -
D
- glycero - -
D
ido-heptonate (14aa).—To a solution of 13aa
(436 mg, 0.368 mmol) in CH2Cl2 (3 mL) were
added DMAP (67.5 mg, 0.553 mmol) and
(PhO)2P(O)Cl (0.115 mL, 0.555 mmol). After
stirring for 2 h at rt, the mixture was diluted
with EtOAc, washed with satd NaHCO3, and
brine, dried over MgSO4, filtered, and chro-
matographed on a silica gel column. Elution
with 3:1 hexane–EtOAc gave 14aa (496 mg,
95%) as a gum. IR wmax(CHCl3): 3429, 3347,
Diphenylmethyl 2,6-anhydro-7-O-tert-butyl-
dimethylsilyl - 3 - deoxy - 4 - O - [(R) - 3 - (dode-
cyloxy)tetradecyl] - 3 - tetradecanamido -
D
-
glycero- -ido-heptonate (13ac).—Compound
D
12ac (448 mg, 0.465 mmol) was treated, as
described in the formation of 13aa from 12aa
to give 13ac (446 mg, 89%) as a gum. IR
wmax(CHCl3): 2928, 2855, 1728, 1672 cm−1. H
2928, 2856, 1754, 1668 cm−1. H NMR (400
1
1
NMR (400 MHz, CDCl3): l 0.06 (3 H, s), 0.07
(3 H, s), 0.84–0.89 (18 H, m), 1.23–1.51 (62
H, m), 1.67–1.72 (2 H, m), 2.04 (2 H, t, J
6.6–8.8 Hz), 3.33–3.44 (5 H, m, containing
OH, 2 H, t, J 6.6 Hz, at l 3.35), 3.54 (1 H, t,
J 8.8–9.5 Hz), 3.65–3.71 (3 H, m), 3.77 (1 H,
dd, J 5.1, 11.0 Hz), 3.83 (1 H, dd, J 4.4, 10.3
Hz), 4.37 (1 H, m), 4.64 (1 H, d, J 5.1 Hz),
6.08 (1 H, d, J 8.8 Hz, NH), 6.89 (1 H, s),
7.29–7.37 (10 H, m). FABMS, (positive-ion):
m/z 1078 (M+H)+. HRFABMS (positive-
ion): Calcd for C66H116NO8Si: 1078.8470;
MHz, CDCl3): l 0.020 (6 H, s), 0.85–0.92 (18
H, m), 1.17–1.47 (58 H, m), 1.59–1.71 (4 H,
m), 2.06 (1 H, dd, J 3.7, 12.5 Hz), 2.18 (1 H,
dd, J 8.1, 15.4 Hz), 3.24–3.39 (4 H, m),
3.65–3.92 (5 H, m), 4.02 (1 H, m), 4.27, 4.36
(2 H, ABq, J 11.4 Hz), 4.59 (1 H, m), 4.66–
4.72 (2 H, m), 6.85 (1 H, s), 6.86 (1 H, d, J 8.8
Hz, NH), 7.11–7.32 (25 H, m). FABMS, (pos-
itive-ion): m/z 1416 (M+H)+. HRFABMS
(positive-ion): Calcd for C85H131NO12PSi:
1416.9178; Found: 1416.9197.
Diphenylmethyl 2,6-anhydro-7-O-tert-butyl-
dimethylsilyl - 3 - deoxy - 3 - (2,2 - difluorotetra-
decanamido) - 5 - O - diphenylphosphono - 4 - O-
Found:
1078.8462.
Anal.
Calcd
for
C66H115NO8Si (1078.7): C, 73.49; H, 10.75; N,
1.30. Found: C, 73.69; H, 10.58; N, 1.37.
[(R) - 3 - (dodecyloxy)tetradecyl] -
D
- glycero - -
D
Diphenylmethyl
2,6-anhydro-4-O-[(R)-3-
ido-heptonate (14ab).—Compound 13ab (368
mg, 0.330 mmol) was treated as described in
the formation of 14aa from 13aa to give 14ab
(benzyloxy)tetradecyl] - 7 - O - tert - butyldime-
thylsilyl - 3 - deoxy - 3 - [(R) - 3 - (dodecyloxy)-

270
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
15aa (385 mg, 93%) as an amorphous residue.
IR wmax(CHCl3): 3433, 2928, 2855, 1731, 1672
(425 mg, 96%) as an oil. IR wmax(CHCl3):
1
3691, 3425, 2928, 2856, 1755, 1706 cm-1. H
1
cm−1. H NMR (400 MHz, CDCl3): l 0.88 (9
NMR (400 MHz, CDCl3): l 0.011 (3 H, s),
0.020 (3 H, s), 0.85–0.91 (18 H, m), 1.17–1.49
(60 H, m), 1.68–1.83 (4 H, m), 3.26 (1 H, m),
3.34 (2 H, t, J 6.6 Hz), 3.67–3.72 (3 H, m),
3.83 (1 H, dd, J 6.6, 10.3 Hz), 3.89 (1 H, dd,
J 6.6, 10.3 Hz), 4.12 (1 H, m), 4.56 (1 H, m),
4.67 (1 H, d, J 2.9 Hz), 4.73 (1 H, m), 6.97 (1
H, s), 7.07 (1 H, d, J 9.5 Hz, NH), 7.16–7.36
(20 H, m). FABMS, (positive-ion): m/z 1384
(M+K)+ (on addition of KI). FABMS, (neg-
ative-ion): m/z 1344 (M−H)−. HRFABMS
(negative-ion): Calcd for C78H121F2NO11PSi:
1344.8415; Found: 1344.8405. Anal. Calcd for
C78H122F2NO11PSi (1346.9): C, 69.56; H, 9.13;
F, 2.82; N, 1.04; P, 2.30. Found: C, 69.55; H,
9.04; F, 2.82; N, 1.09; P, 2.05.
H, t, J 5.9–7.3 Hz), 1.24–1.61 (62 H, m), 2.26
(2 H, d, J 5.9 Hz), 3.15 (1 H, m), 3.22 (2 H, t,
J 6.6–7.3 Hz), 3.40 (1 H, m), 3.50–3.57 (4 H,
m), 3.66 (1 H, m), 3.78 (1 H, dd, J 8.1, 9.5
Hz), 4.31, 4.35 (2 H, ABq, J 11.7 Hz), 4.49 (1
H, m), 4.57 (1 H, q, J 8.8 Hz), 4.73 (1 H, d, J
5.1 Hz), 6.82 (1 H, s), 6.83 (1 H, d, J 7.3 Hz,
NH), 7.12–7.34 (25 H, m). FABMS, (positive-
ion): m/z 1324 (M+Na)+, 1302 (M+H)+.
HRFABMS (positive-ion): Calcd for C79H116-
NNaO12P: 1324.8133; Found: 1324.8129.
Anal. Calcd for C79H116NO12P (1302.7): C,
72.84; H, 8.98; N, 1.08; P, 2.38. Found: C,
72,53; H, 8.72; N, 1.04; P, 1.99.
Diphenylmethyl
2,6-anhydro-3-deoxy-3-
Diphenylmethyl 2,6-anhydro-7-O-tert-butyl-
dimethylsilyl-3-deoxy-5-O-diphenylphosphono-
4-O-[(R)-3-(dodecyloxy)tetradecyl] -3-tetra-
(2,2 - difluorotetradecanamido) - 5 - O - diphenyl-
phosphono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-
D
-glycero-
D
-ido-heptonate
(15ab).—Com-
decanamido-
D
-glycero-
D
-ido-heptonate (14ac).
pound 14ab (390 mg, 0.290 mmol) was treated
as described in the formation of 15aa from
14aa to give 14ab (316 mg, 88%) as an amor-
phous solid. IR wmax(KBr): 3449, 3338, 2922,
—Compound 13ac (425 mg, 0.394 mmol) was
treated, as described in the formation of 14aa
from 13aa to give 14ac (500 mg, 97%) as a
gum. IR wmax(CHCl3): 2928, 2855, 1752, 1675
2853, 1713 cm−1
.
1H NMR (400 MHz,
1
cm−1. H NMR (400 MHz, CDCl3): l 0.07 (3
CDCl3): l 0.88 (9 H, t, J 6.6 Hz), 1.18–1.45
(60 H, m), 1.63–1.66 (2 H, m), 1.94–1.98 (2
H, m), 3.18 (1 H, m), 3.24–3.35 (4 H, m,
containing OH), 3.58–3.66 (3 H, m), 3.72–
3.82 (2 H, m), 4.50 (1 H, m), 4.60 (1 H, dd, J
8.1, 17.6 Hz), 4.68 (1 H, d, J 5.1 Hz), 6.95 (1
H, s), 7.02 (1 H, d, J 9.5 Hz, NH), 7.13–7.37
(20 H, m). FABMS, (positive-ion): m/z 1254
(M+Na)+, 1232 (M+H)+. HRFABMS
(positive-ion): Calcd for C72H108NNaO11F2P:
1254.7526; Found: 1254.7517.
H, s), 0.018 (3 H, s), 0.83–0.89 (18 H, m),
1.11–1.48 (62 H, m), 1.68–1.82 (4 H, m), 3.28
(1 H, m), 3.34 (2 H, t, J 6.6 Hz), 3.66–3.85 (4
H, m), 3.93 (1 H, dd, J 2.9 Hz), 4.20 (1 H, m),
4.55 (1 H, m), 4.67 (1 H, d, J 2.9 Hz), 4.73 (1
H, m), 6.00 (1 H, d, J 9.5 Hz, NH), 6.94 (1 H,
s), 7.16–7.35 (20 H, m). FABMS, (positive-
ion): m/z 1332 (M+Na)+, 1310 (M+H)+.
HRFABMS (positive-ion): Calcd for C78H124-
NNa:O11PSi 1332.8579; Found: 1332.8627.
Anal. Calcd for C78H124NO11PSi (1310.9): C,
71.47; H, 9.54; N, 1.07; P, 2.36. Found: C,
71.59; H, 9.59; N, 1.38; P, 2.46.
Diphenylmethyl 2,6-anhydro-3-deoxy-5-O-
diphenylphosphono-4-O-[(R)-3-(dodecyloxy)-
tetradecyl] - 3 - tetradecanamido -
D
- glycero - -
D
Diphenylmethyl 2,6-anhydro-3-[(R)-3-(ben-
zyloxy)tetradecanamido] - 3 - deoxy - 5 - O-
diphenylphosphono-4-O-[(R)-3-(dodecyloxy)-
ido-heptonate (15ac).—Compound 14ac (471
mg, 0.359 mmol) was treated as described in
the formation of 15aa from 14aa to give 15ac
(392 mg, 91%) as a white solid; mp 73.5–
74.5 °C. [h]2D5 +15.4° (c 0.15, CHCl3). IR
wmax(CHCl3): 3435, 2927, 2855, 1728, 1677
tetradecyl]-
D
-glycero- -ido-heptonate (15aa).
D
—A solution of 14aa (452 mg, 0.319 mmol) in
THF (3 mL) and 3 M HCl aq (0.4 mL) was
stirred for 6 h at rt. The reaction mixture was
diluted with EtOAc, washed with satd
NaHCO3 and brine, dried over MgSO4,
filtered, and chromatographed on a silica gel
column. Elution with 1:1 hexane–EtOAc gave
1
cm−1. H NMR (400 MHz, CDCl3): l 0.88 (9
H, t, J 6.6–7.3 Hz), 1.20–1.47 (60 H, m),
1.62–1.64 (4 H, m), 1.92–1.98 (2 H, m), 3.15
(1 H, brs, OH), 3.26–3.29 (3 H, m, containing
2 H, t, J 6.6 Hz, at 3.27), 3.59–3.67 (4 H, m),

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
271
NMR (400 MHz, CDCl3): l 0.88 (9 H, t, J
6.6–7.3 Hz), 1.10–1.68 (62 H, m), 2.20–2.32
(2 H, m), 3.28–3.31 (3 H, m, containing 2 H,
t, J 6.6 Hz, at 3.30), 3.66–3.75 (4 H, m), 3.84
(1 H, t, J 6.6–7.3 Hz), 3.97–3.99 (2 H, m),
4.49 (1 H, m), 4.59 (1 H, m), 4.70 (1 H, d, J
4.4 Hz), 7.18–7.35 (10 H, m). FABMS, (posi-
tive-ion): m/z 1068 (M+Na)+, 1046 (M+
H)+. HRFABMS (positive-ion): Calcd for
3.77–3.82 (2 H, m), 4.49 (1 H, m), 4.59 (1 H,
m), 4.78 (1 H, d, J 5.1 Hz), 6.04 (1 H, d, J 8.8
Hz, NH), 6.91 (1 H, s), 7.15–7.35 (20 H, m).
FABMS, (positive-ion): m/z 1218 (M+Na)+,
1196 (M+H)+. HRFABMS (positive-ion):
Calcd for C72H110NNaO11P: 1218.7714;
Found: 1218.7747. Anal. Calcd for C72H110-
NO11P (1196.6): C, 72.27; H, 9.27; N, 1.17; P,
2.59. Found: C, 71.88; H, 8.85; N, 1.18; P,
2.56.
C59H100NNaO12P:
1068.6887.
1068.6881;
Found:
Diphenylmethyl
2,6-anhydro-4-O-[(R)-3-
(benzyloxy)tetradecyl]-3-deoxy-5-O-diphenyl-
phosphono - 3 - [(R) - 3 - (dodecyloxy)tetradecan-
2,6-Anhydro-3-deoxy-3-(2,2-difluorotetra-
decanamido) - 5 - O - diphenylphosphono - 4 - O-
amido]-
D
-glycero-
D
-ido-heptonate (15ba).—
[(R) - 3 - (dodecyloxy)tetradecyl] -
D
- glycero - -
D
To a solution of 13ba (200 mg, 0.168 mmol) in
CH2Cl2 (2 mL) were added DMAP (41.2 mg,
0.338 mmol) and (PhO)2P(O)Cl (0.07 mL,
0.338 mmol). After stirring for 2 h at rt, the
reaction mixture was diluted with EtOAc,
washed with satd NaHCO3 aq, and brine,
dried over MgSO4, filtered, and concentrated
to give a mixture of 14ba and spent reagents.
This mixture was treated, as described in the
formation of 15aa from 14aa to give 15ba (176
mg, 80%). IR wmax(CHCl3): 3467, 2920, 2851,
ido-heptonic acid (16ab).—Compound 15ab
(42.5 mg, 0.035 mmol) was treated as de-
scribed below in the formation of 16ac from
15ac to give 16ab (25.4 mg, 69%) as an amor-
phous solid. IR wmax(CHCl3): 3381, 3309,
1
2922, 2851, 1717, 1691 cm−1. H NMR (400
MHz, 9:1 CD3OD–CDCl3): l 0.89 (9 H, t, J
6.6 Hz), 1.13–1.48 (60 H, m), 1.66–1.71 (2 H,
m), 2.03–2.07 (2 H, m), 3.27–3.36 (3 H, m),
3.73–3.77 (4 H, m), 3.87 (1 H, t, J 6.6–7.3
Hz), 4.07 (1 H, m), 4.44 (1 H, m), 4.60 (1 H,
d, J 4.4 Hz), 4.67 (1 H, m), 7.20–7.40 (10 H,
m), 8.01 (1 H, d, J 8.8 Hz, NH). FABMS,
(positive-ion): m/z 1088 (M+Na)+, 1066
(M+H)+. HRFABMS (positive-ion): Calcd
for C59H98F2NNaO11P: 1088.6743; Found:
1088.6752.
1733, 1649 cm−1
.
1H NMR (400 MHz,
CDCl3): l 0.88(9 H, t, J 6.8–7.0 Hz), 1.17–
1.77 (6 H, m), 2.14 (2 H, d, J 6.1 Hz),
3.13–3.17 (1 H, m), 3.21–3.31 (2 H, m), 3.38–
3.42 (2 H, m), 3.55 (2 H, s), 3.73–3.82 (2 H,
m), 3.86–3.90 (1 H, m), 4.37 (2 H, s), 4.53 (1
H, dt, J 5.4, 9.1 Hz), 4.60 (1 H, dd, J 8.2, 17.4
Hz), 4.72 (1 H, d, J 5.3 Hz), 6.88 (1 H, s), 6.95
(1 H, d, J 9.0 Hz), 7.13–7.34 (25 H, m).
2,6 - Anhydro - 3 - deoxy - 5 - O - diphenylphos-
phono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-3-
tetradecanamido-
D
-glycero- -ido-heptonic acid
D
HRFABMS,
(positive-ion):
Calcd
for
(16ac).—A solution of 15ac (38.1 mg, 0.032
mmol) in EtOH (2 mL) containing 10% Pd/C
(18.8 mg) was stirred vigorously under N2 for
2 h at rt. The reaction mixture was filtered
and concentrated in vacuo to give a crude
product. The crude product was chro-
matographed on a short column of silica gel.
Elution with 8:1 CHCl3–MeOH gave 16ac
(25.6 mg, 78%) as an amorphous residue. IR
wmax(CHCl3): 3434, 2927, 2855, 1747, 1675
C79H117NO12P (M+H)+: 1302.8313; Found:
1302.8339.
2,6 - Anhydro - 3 - deoxy - 5 - O - diphenylphos-
phono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-3-
[(R)-3-hydroxytetradecanamido]-
D
-glycero- -
D
ido-heptonic acid (16aa).—A solution of 15aa
(64.5 mg, 0.050 mmol) in EtOH (2 mL) con-
taining 20% Pd(OH)2-on-carbon (31 mg) was
stirred vigorously under hydrogen for 8 h at
rt. The reaction mixture was filtered and con-
centrated in vacuo to give a crude product.
The crude product was chromatographed on a
silica gel short solumn. Elution with 8:1
CHCl3–MeOH gave 16aa (42.3 mg, 82%) as
an amorphous residue. IR wmax(KBr): 3334,
1
cm−1. H NMR (400 MHz, CDCl3): l 0.88 (9
H, t, J 6.6–7.3 Hz), 1.11–1.56 (62 H, m),
1.66–1.68 (2 H, m), 2.08–2.15 (2 H, m), 3.30–
3.33 (3 H, m, containing 2 H, t, J 6.6–7.3 Hz,
at 3.32), 3.68–3.83 (5 H, m), 3.95 (1 H, m),
4.52 (1 H, m), 4.60 (1 H, m), 4.67 (1 H, d, J
3.7 Hz), 6.50 (1 H, d, J 8.8 Hz, NH), 7.19–
1
3063, 2955, 2922, 2852, 1742, 1649 cm−1. H

272
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
m), 4.57 (1 H, d, J 4.4 Hz). FABMS, (posi-
tive-ion): m/z 916 (M+Na)+. FABMS, (neg-
ative-ion): m/z 892 (M−H)−. HRFABMS
(positive-ion): Calcd for C47H92NO12PNa:
916.6225; Found: 916.6227. Anal. Calcd For
C47H92NO12P (894.2): C. 63.13; H, 10.37; N,
1.57; P, 3.46. Found: C, 62.12; H, 9.91; N,
1.58; P, 3.35.
7.37 (10 H, m). FABMS, (positive-ion): m/z
1052 (M+Na)+. HRFABMS (positive-ion):
Calcd for C59H100NNaO11P: 1052.6932;
Found: 1052.6930.
2,6 - Anhydro - 3 - deoxy - 5 - O - diphenylphos-
phono-3-[(R)-3-(dodecyloxy)tetradecanamido]-
4-O-[(R)-3-hydroxytetradecyl]-
D
-glycero- -
D
ido-heptonic acid (16ba).—A solution of 15ba
(26.0 mg, 0.020 mmol) in EtOH (2 mL) con-
taining 20% Pd(OH)2-on-carbon (30.0 mg)
was stirred vigorously under hydrogen for 16
h at rt. The reaction mixture was filtered and
concentrated in vacuo to give a crude product.
The crude product was chromatographed on a
silica gel TLC plate. Elution with 4:1 CHCl3–
MeOH gave 16ba (12.2 mg, 58%). IR
2,6-Anhydro-3-deoxy-3-(2,2-difluorotetra-
decanamido) - 4 - O - [(R) - 3 - (dodecyloxy)tetra-
decyl]-5-O-phosphono-
D
-glycero- -ido-heptonic
D
acid (17ab).—Compound 16ab (21.5 mg,
0.020 mmol) was treated as described in the
formation of 17aa from 16aa to give 17ab
(17.8 mg, 96%) as a white powder; mp 133.0–
135.5 °C. [h]2D3 +4.1° (c 0.60, CHCl3). IR
wmax(KBr): 3338, 2922, 2852, 1688 cm−1. H
1
1
wmax(KBr): 3412, 2919, 2850, 1645 cm−1. H
NMR (500 MHz, CDCl3): l 0.88–0.91 (m, 9
H), 1.19–1.70 (m, 62 H), 2.32–2.45 (m, 2 H),
3.31 (s, 3 H), 3.41–3.44 (m, 1 H), 3.48–3.54
(m, 1 H), 3.60–3.62 (m, 1 H), 3.65–3.76 (m, 4
H), 3.83 (dd, 1 H, J 6.2 Hz, 8.2 Hz), 3.87–3.94
(m, 2 H), 4.32 (d, 1 H, J 4.5 Hz), 4.37–4.41
(m, 1 H), 4.61 (dd, 1 H, J 1.8, 7.4 Hz),
7.21–7.40 (m, 10 H). HRFABMS (positive-
ion): Calcd for C59H101NO12P (M+H)+:
1046.7061; Found:1046.7063.
NMR (400 MHz, 5:1 CD3OD–CDCl3): l 0.89
(9 H, t, J 6.6–7.3 Hz), 1.20–1.58 (60 H, m),
1.74–1.79 (2 H, m), 1.99–2.08 (2 H, m), 3.39
(1 H, m), 3.45 (2 H, t, J 6.6 Hz), 3.70–3.76 (3
H, m), 3.82 (1 H, m), 3.94–3.97 (2 H, m), 4.27
(1 H, m), 4.42 (1 H, dd, J 4.4, 6.6 Hz), 4.59 (1
H, d, J 3.7 Hz). FABMS, (positive-ion): m/z
936 (M+Na)+, 914 (M+H)+. FABMS,
(negative-ion): m/z 912 (M−H)−. HR-
FABMS (positive-ion): Calcd for C47H90-
NNaO11F2P: 936.6117; Found: 936.6133.
Anal. Calcd for C47H90NO11F2P (914.2): C,
61.75; H, 9.92; N, 1.53; F, 4.16; P, 3.39.
Found: C, 61.70; H, 9.51; N, 1.38; F, 3.88; P,
3.02.
2,6-Anhydro-3-deoxy-4-O-[(R)-3-(dode-
cyloxy)tetradecyl] - 3 - [(R) - 3 - hydroxytetra-
decanamido]-5-O-phosphono-
D
-glycero- -ido-
D
heptonic acid (17aa).—A solution of 16aa
(37.5 mg, 0.036 mmol) in THF (2 mL) con-
taining PtO2 (20.1 mg) was stirred vigorously
under hydrogen for 18 h at rt. The reaction
mixture was filtered and concentrated in
vacuo to give a residue. The residue was dis-
solved in CHCl3 (4 mL), MeOH (8 mL) and
aq 0.1M HCl (4 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 concentrated to give 17aa (23.8
mg, 74%) as a white powder; mp. 129.5–
130.5 °C. [h]2D4 +11.2° (c 0.15, CHCl3). IR
wmax(KBr): 3313, 2956, 2922, 2853, 1734, 1646
2,6-Anhydro-3-deoxy-4-O-[(R)-3-(dode-
cyloxy)tetradecyl] - 5 - O - phosphono - 3 - tetra-
decanamido-
D
-glycero-
D
-ido-heptonic
acid
(17ac).—Compound 16ac (23.8 mg, 0.023
mmol) was treated as described in the forma-
tion of 17aa from 16aa to give 17ac (20.0 mg,
99%) as a white powder; mp. 119.5–121.0 °C.
[h]2D6 +8.3° (c 0.41, CHCl3). IR wmax(KBr):
1
3307, 2922, 2852, 1733, 1650 cm−1. H NMR
(400 MHz, 9:1 CD3OD–CDCl3): l 0.90 (9 H,
t, J 6.6–7.3 Hz), 1.20–1.78 (64 H, m), 2.23 (2
H, t, J 7.3–8.1 Hz), 3.42–3.48 (3 H, m, con-
taining 2 H, t, J 6.6 Hz, at l 3.46), 3.70–3.81
(4 H, m), 3.90 (1 H, m), 4.04 (1 H, m), 4.20 (1
H, m), 4.34 (1 H, m), 4.56 (1 H, d, J 4.4 Hz).
FABMS, (positive-ion): m/z 900 (M+Na)+.
1
cm−1. H NMR (400 MHz, 5:1 CD3OD–
CDCl3): l 0.88 (9 H, t, J 6.6–7.3 Hz), 1.16–
1.58 (60 H, m), 1.75–1.78 (2 H, m), 2.31 (1 H,
dd, J 8.1, 14.6 Hz), 2.39 (1 H, dd, J 4.4, 14.6
Hz), 3.42–3.49 (3 H, m), 3.71–3.80 (4 H, m),
3.90–3.99 (3 H, m), 4.22 (1 H, m), 4.39 (1 H,
HRFABMS
(positive-ion):
Calcd
for
C47H92NNaO11P: 900.6306; Found: 900.6315.
Anal. Calcd for C47H92NO11P (878.2): C,

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
273
FABMS, (positive-ion): m/z 1316 (M+H)+.
HRFABMS (positive-ion): Calcd for
C80H119NO12P: 1316.8470; Found: 1316.8475.
Diphenylmethyl 2,6-anhydro-3-deoxy-3-
(2,2 - difluorotetradecanamido) - 5 - O - diphenyl-
phosphono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-
64.28; H, 10.56; N, 1.60; P, 3.53. Found: C,
62.62; H, 10.45; N, 1.24; P, 3.45.
2,6-Anhydro-3-deoxy-3-[(R)-3-(dodecyloxy)-
tetradecanamido] - 4 - O - [(R) - 3 - hydroxytetra-
decyl] - 5 - O - phosphono -
D
- glycero -
D
- ido-
heptonic acid (17ba).—A solution of 16ba (8.7
mg, 0.008 mmol) in THF (2 mL) containing
PtO2 (21.3 mg, 0.094 mmol) was stirred vigor-
ously under hydrogen 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 (4 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 concentrated to give
17ba (6.4 mg, 86%). [h]2D4 −6.9° (c 0.30,
CHCl3). IR wmax(KBr): 3370, 2957, 2919, 2850,
7-O-methyl-
D
-glycero- -ido-heptonate (18b).
D
—Compound 15ab (78.9 mg, 0.064 mmol)
was treated as described in the formation of
18a from 15aa to give 18b (56.1 mg, 70%) as
an amorphous solid. IR wmax(CHCl3): 2927,
2855, 1709 cm−1
.
1H NMR (400 MHz,
CDCl3): l 0.88 (9 H, t, J 5.9–7.3 Hz), 1.20–
1.46 (60 H, m), 1.66 (2 H, q, J 6.6 Hz),
1.69–1.83 (2 H, m), 3.22 (1 H, m), 3.27 (3 H,
s), 3.29 (2 H, t, J 7.3 Hz), 3.52–3.70 (5 H, m),
4.01 (1 H, m), 4.53 (1 H, m), 4.63 (1 H, d, J
5.1 Hz), 4.67 (1 H, d, J 4.4 Hz), 6.96 (1 H, s),
7.03 (1 H, d, J 8.8 Hz, NH), 7.16–7.37 (20 H,
m). FABMS, (positive-ion): m/z 1268 (M+
Na)+, 1246 (M+H)+. HRFABMS (positive-
1
1646 cm−1. H NMR (400 MHz, CD3OD): l
0.90 (9 H, t, J 6.8 Hz), 1.16–1.78 (62 H, m),
2.28–2.30 (2 H, m), 3.41–3.48 (1 H, m), 3.50–
3.56 (1 H, m), 3.66–3.69 (2 H, m), 3.73–3.92
(6 H, m), 4.24–4.25 (1 H, m), 4.33–4.35 (1 H,
m), 4.52 (1 H, d, J 4.9 Hz). HRFABMS
(positive-ion): Calcd for C49H93NO12P (M+
H)+: 894.6435; Found: 894.6436.
ion):
Calcd
for
C73H110NNaO11F2P:
1268.7682; Found: 1268.7653.
Diphenylmethyl 2,6-anhydro-3-deoxy-5-O-
diphenylphosphono-4-O-[(R)-3-(dodecyloxy)-
tetradecyl]-7-O-methyl-3-tetradecanamido-
glycero- -ido-heptonate (18c).—Compound
D
-
D
Diphenylmethyl 2,6-anhydro-3-[(R)-3-(ben-
zyloxy)tetradecanamido] - 3 - deoxy - 5 - O-
diphenylphosphono-4-O-[(R)-3-(dodecyloxy)-
15ac (73.2 mg, 0.061 mmol) was treated as
described in the formation of 18c from 15ac to
give 18c (64.1 mg, 87%) as a white solid; mp.
70.5–71.0 °C. [h]D26 +4.4° (c 0.18, CHCl3). IR
tetradecyl] - 7 - O - methyl -
D
- glycero -
D
- ido-
heptonate (18a).—To a solution of 15aa (86.0
wmax(KBr): 3349, 2922, 2852, 1741, 1648 cm−1
.
mg, 0.066 mmol) in CH2Cl2 (3 mL) were
1H NMR (400 MHz, CDCl3): l 0.88 (9 H, t, J
6.6 Hz), 1.13–1.47 (62 H, m), 1.65–1.70 (2 H,
m), 1.77–1.83 (2 H, m), 3.26 (1 H, m), 3.29 (3
H, s), 3.32 (2 H, t, J 6.6 Hz), 3.56 (1 H, dd, J
5.9, 10.3 Hz), 3.77 (1 H, t, J 5.1 Hz), 4.16 (1
H, m), 4.53 (1 H, m), 4.63 (1 H, m), 4.72 (1 H,
d, J 3.7 Hz), 5.98 (1 H, d, J 8.8 Hz, NH), 6.93
(1 H, s), 7.17–7.37 (20 H, m). FABMS, (posi-
tive-ion): m/z 1232 (M+Na)+, 1210 (M+
H)+. HRFABMS (positive-ion): Calcd for
added
2,6-di-tert-butyl-4-methyl-pyridine
(44.8 mg, 0.218 mmol) and Me3OBF4 (30.1
mg, 0.204 mmol). After stirring for 3 h at rt,
the mixture was diluted with EtOAc, washed
with satd NaHCO3 and brine, dried over
MgSO4, filtered, and chromatographed on a
silica gel column. Elution with 7:3 hexane–
EtOAc gave 18a (80.2 mg, 92%) as an amor-
phous residue. IR wmax(CHCl3): 3691, 2928,
1
2855, 1732, 1669 cm−1. H NMR (400 MHz,
C73H112NNaO11P:
1232.7871.
1232.7833;
Found:
CDCl3): l 0.88 (9 H, t, J 5.9–7.3 Hz), 1.24–
1.66 (62 H, m), 2.11 (1 H, dd, J 3.7, 15.4 Hz),
2.20 (1 H, dd, J 8.1, 15.4 Hz), 3.19–3.29 (6 H,
m, containing 3 H, s, at l 3.26), 3.44–3.64 (5
H, m), 3.78 (1 H, m), 3.94 (1 H, m), 4.28, 4.38
(2 H, ABq, J 11.7 Hz), 4.53–4.63 (2 H, m),
4.70 (1 H, d, J 4.4 Hz), 6.84 (1 H, s), 6.85 (1
H, d, J 8.8 Hz, NH), 7.12–7.33 (25 H, m).
2,6 - Anhydro - 3 - deoxy - 5 - O - diphenylphos-
phono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-3-
[(R)-3-hydroxytetradecanamido]-7-O-methyl-
D
-glycero- -ido-heptonic acid (19a).—Com-
D
pound 18a (75.5 mg, 0.0573 mmol) was
treated as described in the formation of 16aa
from 15aa to give 19a (50.3 mg, 83%) as an

274
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
amorphous residue. IR wmax(KBr): 3343, 3069,
2,6-Anhydro-3-deoxy-4-O-[(R)-3-(dode-
cyloxy)tetradecyl] - 3 - [(R) - 3 - hydroxytetra-
1
2922, 2852, 1734, 1651 cm−1. H NMR (400
MHz, CDCl3): l 0.88 (9 H, t, J 6.6–7.3 Hz),
1.25–1.53 (60 H, m), 1.67–1.72 (2 H, m), 2.15
(1 H, dd, J 9.5, 15.4 Hz), 2.26 (1 H, m),
3.26–3.38 (6 H, m, containing 3 H, s, at l
3.31), 3.59–3.78 (6 H, m), 3.94 (1 H, m), 4.31
(1 H, m), 4.58 (1 H, d, J 4.4 Hz), 4.65 (1 H,
m), 7.06 (1 H, d, J 9.5 Hz, NH), 7.18–7.37 (10
H, m). FABMS, (positive-ion): m/z 1082
(M+Na)+, 1060 (M+H)+. HRFABMS
(positive-ion): Calcd for C60H102NNaO12P:
1082.7037; Found: 1082.7019.
decanamido]-7-O-methyl-5-O-phosphono-
D
-
glycero- -ido-heptonic acid (20a).—Com-
D
pound 19a (46.5 mg, 0.0439 mmol) was
treated as described in the formation of 17aa
from 16aa to give 20a (36.7 mg, 92%) as a
white powder; mp 102.5–103.0 °C. [h]2D5 +6.7°
(c 0.14, CHCl3). IR wmax(KBr): 3310, 2956,
1
2923, 2853, 1733, 1648 cm−1. H NMR (400
MHz, 5:1 CD3OD–CDCl3): l 0.88 (9 H, t, J
6.6–7.3 Hz), 1.18–1.58 (60 H, m), 1.76–1.77
(2 H, m), 2.31 (1 H, dd, J 8.1, 14.6 Hz), 2.39
(1 H, dd, J 4.4, 14.6 Hz), 3.32–3.49 (6 H, m,
containing 3 H, s, at l 3.39), 3.65–3.78 (5 H,
m), 3.93 (1 H, m), 4.19–4.27 (2 H, m), 4.43 (1
H, m), 4.57 (1 H, d, J 3.7 Hz). FABMS,
(positive-ion): m/z 930 (M+Na)+. FABMS,
(negative-ion): m/z 906 (M−H)−. HR-
FABMS (positive-ion): Calcd for C48H94-
NNaO12P: 930.6411; Found: 930.6417. Anal.
Calcd for C48H94NO12P (908.2): C, 63.48; H,
10.43; N, 1.54; P, 3.41. Found: C, 61.81; H,
10.45; N, 1.49; P, 3.45.
2,6-Anhydro-3-deoxy-3-(2,2-difluorotetra-
decanamido) - 5 - O - diphenylphosphono - 4 - O-
[(R)3-(dodecyloxy)tetradecyl]-7-O-methyl-
D
-
glycero- -ido-heptonic acid (19b).—Com-
D
pound 15ab (39.5 mg, 0.032 mmol) was
treated as described in the formation of 16ac
from 15ac to give 19b (23.6 mg, 69%) as an
amorphous solid. IR wmax(KBr): 3352, 2922,
1
2852, 1733, 1695 cm−1. H NMR (400 MHz,
CDCl3): l 0.88 (9 H, t, J 6.6 Hz), 1.23–1.49
(60 H, m), 1.69 (2 H, q, J 6.6 Hz), 1.98–2.05
(2 H, m), 3.25–3.35 (6 H, m, containing 3 H,
s, at l 3.30), 3.52 (1 H, dd, J 4.4, 11.0 Hz),
3.67–3.75 (4 H, m), 4.23 (1 H, m), 4.54–4.62
(3 H, m, containing 1 H, d, J 3.7 Hz, at l
4.60), 7.18–7.36 (11 H, m, containing NH).
FABMS, (positive-ion): m/z 1102 (M+Na)+,
1080 (M+H)+. HRFABMS (positive-ion):
Calcd for C60H100F2NNaO11P: 1102.6900;
Found: 1102.6884.
2,6-Anhydro-3-deoxy-3-(2,2-difluorotetra-
decanamido) - 4 - O - [(R) - 3 - (dodecyloxy)tetra-
decyl]-7-O-methyl-5-O-phosphono-
-ido-heptonic acid (20b).—Compound 19b
D
-glycero-
D
(20.8 mg, 0.0193 mmol) was treated as de-
scribed in the formation of 17aa from 16aa to
give 20b (17.6 mg, 98%) as a white powder;
mp 124.5–125.5 °C. [h]2D6 +3.0° (c 0.65,
CHCl3). IR wmax(KBr): 3348, 2923, 2853, 1718,
2,6 - Anhydro - 3 - deoxy - 5 - O - diphenylphos-
phono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-7-
1693 cm−1
.
1H NMR (400 MHz, 9:1
CD3OD–CDCl3): l 0.90 (9 H, t, J 5.9–7.3
Hz), 1.20–1.56 (60 H, m), 1.76–1.77 (2 H, m),
2.01–2.07 (2 H, m), 3.39 (3 H, s), 3.42–3.48 (3
H, m), 3.66–3.80 (5 H, m), 4.24–4.31 (2 H,
m), 4.44 (1 H, m), 4.84 (1 H, d, J 3.7 Hz).
FABMS, (positive-ion): m/z 950 (M+Na)+.
O-methyl-3-tetradecanamido-
D
-glycero- -ido-
D
heptonic acid (19c).—Compound 18c (58.1
mg, 0.0480 mmol) was treated as described in
the formation of 16ac from 15ac to give 19c
(31.2 mg, 62%) as an amorphous residue. IR
wmax(CHCl3): 3436, 2927, 2855, 1774, 1753,
1
1677 cm−1. H NMR (400 MHz, CDCl3): l
HRFABMS
(positive-ion):
Calcd
for
C48H92NO11F2PNa:
950.6274;
Found:
0.88 (9 H, t, J 6.6–7.3 Hz), 1.20–1.52 (62 H,
m), 1.67–1.72 (2 H, m), 2.00–2.17 (2 H, m),
3.26–3.37 (6 H, m, containing 3 H, s, at l
3.31), 3.59 (1 H, m), 3.66–3.75 (4 H, m), 4.31
(1 H, m), 4.57–4.62 (3 H, m, containing 1 H,
d, J 4.4 Hz, at l 4.61), 6.49 (1 H, d, J 8.8 Hz,
NH), 7.20–7.36 (10 H, m). FABMS, (positive-
ion): m/z 1066 (M+Na)+. HRFABMS (posi-
950.6270. Anal. Calcd for C48H92F2NO11P
(928.2): C, 62.11; H, 9.99; F, 4.09; N, 1.51; P,
3.34. Found: C, 61.77; H, 10.14; F, 3.84; N,
1.54; P, 3.61.
2,6-Anhydro-3-dzeoxy-4-O-[(R)-3-(dode-
cyloxy)tetradecyl]-7-O-methyl-5-O-phosphono-
3-tetradecanamido-
D
-glycero- -ido-heptonic
D
acid (20c).—Compound 19c (28.6 mg, 0.027
tive-ion):
Calcd
for
C60H102NNaO11P:
1066.7088; Found: 1066.7092.
mmol) was treated as described in the forma-

Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
275
mixture was quenched with water, extracted
with EtOAc, washed with satd 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 21b (45.1 mg,
77%) as an amorphous solid. IR wmax(CHCl3):
tion of 17aa from 16aa to give 19c (22.5 mg,
92%) as a white powder; mp 95.5–96.0 °C.
[h]2D6 +9.4° (c 0.54, CHCl3). IR wmax(KBr):
1
3333, 2922, 2852, 1734, 1650 cm-1. H NMR
(400 MHz, 9:1 CD3OD–CDCl3): l 0.90 (9 H,
t, J 6.6–7.3 Hz), 1.29–1.60 (62 H, m), 1.75–
1.77 (2 H, m), 2.22 (2 H, t, J 7.3–8.1 Hz), 3.38
(3 H, s), 3.42–3.48 (3 H, m), 3.66–3.79 (5 H,
m), 4.23 (1 H, m), 4.39 (1 H, m), 4.56 (1 H, d,
J 3.7 Hz). FABMS, (positive-ion): m/z 914
(M+Na)+, 892 (M+H)+. HRFABMS (pos-
1
3691, 2927, 2855, 1756, 1709 cm−1. H NMR
(400 MHz, CDCl3): l 0.88 (9 H, t, J 6.6 Hz),
1.20–1.47 (60 H, m), 1.66 (2 H, q, J 6.6 Hz),
1.75–1.83 (2 H, m), 3.21 (1 H, m), 3.25–3.37
(2 H, m), 3.65–3.71 (3 H, m), 4.09 (1 H, m),
4.46 (1 H, ddd, J 3.7, 10.3, 46.1 Hz), 4.51–
4.57 (2 H, m), 4.61 (1 H, ddd, J 6.6, 10.3, 46.1
Hz), 4.71 (1 H, d, J 3.7 Hz), 6.97 (1 H, s), 7.02
(1 H, d, J 9.5 Hz, NH), 7.16–7.36 (20 H, m).
FABMS, (positive-ion): m/z 1256 (M+Na)+,
1234 (M+H)+. HRFABMS (positive-ion):
Calcd for C72H107F3NNaO10P: 1256.7482;
Found: 1256.7454.
itive-ion):
Calcd
for
C48H94NNaO11P:
914.6462; Found: 914.6450.
Diphenylmethyl 2,6-anhydro-3-[(R)-3-(ben-
zyloxy)tetradecanamido] - 3,7 - dideoxy - 5 - O-
diphenylphosphono-4-O-[(R)-3-(dodecyloxy)-
tetradecyl]-7-fluoro-
D
-glycero- -ido-heptonate
D
(21a).—To a solution of 15aa (101 mg, 0.078
mmol) in DME (2 mL) was gradually added a
solution of DAST (0.03 mL, 0.227 mmol) in
DME (1 mL) at 0 °C with stirring under N2.
After stirring for 2 h at 0 °C, the reaction
mixture was quenched with water, extracted
with EtOAc, washed with satd NaHCO3 and
brine, dried over MgSO4, filtered, and chro-
matographed on a silica gel column. Elution
with 3:1 hexane–EtOAc gave 21a (52.2 mg,
52%) as an amorphous residue. IR
2,6-Anhydro-3,7-dideoxy-5-O-diphenylphos-
phono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-7-
fluoro-3-[(R)-3-hydroxytetradecanamido] -
glycero- -ido-heptonic acid (22a).—Com-
D
-
D
pound 21a (21.8 mg, 0.017 mmol) was treated
as described in the formation of 16aa from
15aa to give 22a (12.6 mg, 72%) as an amor-
phous residue. IR wmax(KBr): 3350, 2923,
2853, 1736, 1653 cm−1. H NMR (400 MHz,
1
1
wmax(CHCl3): 2928, 2855, 1755, 1670 cm−1. H
NMR (400 MHz, CDCl3): l 0.88 (9 H, t, J
5.9–7.3 Hz), 1.24–1.65 (62 H, m), 2.12 (1 H,
dd, J 3.7, 15.4 Hz), 2.20 (1 H, dd, J 8.1, 15.4
Hz), 3.20 (1 H, m), 3.27 (2 H, t, J 6.6 Hz),
3.41 (1 H, m), 3.60–3.63 (2 H, m), 3.80 (1 H,
t, J 6.6 Hz), 4.00 (1 H, m), 4.29, 4.33 (2 H,
ABq, J 11.0 Hz), 4.36–4.64 (4 H, m), 4.73 (1
H, d, J 4.4 Hz), 6.86 (1 H, s), 6.89 (1 H, d, J
8.8 Hz, NH), 7.11–7.35 (25 H, m). FABMS,
(positive-ion): m/z 1326 (M+Na)+, 1304
(M+H)+. HRFABMS (positive-ion): Calcd
for C79H116FNO11P: 1304.8270; Found:
1304.8263.
CDCl3): l 0.88 (9 H, t, J 6.6 Hz), 1.21–1.51
(60 H, m), 1.68–1.70 (2 H, m), 2.16 (1 H, dd,
J 9.5, 15.4 Hz), 2.27 (1 H, m), 3.30–3.41 (3 H,
m), 3.67–3.77 (4 H, m), 3.94 (1 H, m), 4.46 (1
H, m), 4.57–4.65 (4 H, m), 7.12 (1 H, d, J 9.5
Hz, NH), 7.18–7.38 (10 H, m). FABMS, (pos-
itive-ion): m/z 1070 (M+Na)+, 1048
(M+H)+. HRFABMS (positive-ion): Calcd
for C59H99FNNaO11P: 1070.6838; Found:
1070.6848.
2,6-Anhydro-3,7-dideoxy-3-(2,2-difluorotet-
radecanamido)-5-O-diphenylphosphono-4-O-
[(R)3 - (dodecyloxy)tetradecyl] - 7 - fluoro -
glycero- -ido-heptonic acid (22b).—Com-
D
-
Diphenylmethyl 2,6-anhydro-3,7-dideoxy-3-
(2,2 - difluorotetradecanamido) - 5 - O - diphenyl-
phosphono-4-O-[(R)-3-(dodecyloxy)tetradecyl]-
D
pound 21b (40.5 mg, 0.033 mmol) was treated
as described in the formation of 16ac from
15ac to give 22b (25.1 mg, 72%) as an amor-
phous solid. IR wmax(CHCl3): 3434, 2928,
7-fluoro-
D
-glycero- -ido-heptonate (21b).—
D
To a solution of 15ab (58.4 mg, 0.0474 mmol)
in CH2Cl2 (2 mL) was gradually added a
solution of DAST (0.016 mL, 0.121 mmol) in
CH2Cl2 (1 mL) at −40 °C with stirring under
N2. After stirring for 2 h at 0 °C, the reaction
1
2855, 1780, 1711 cm−1. H NMR (400 MHz,
CDCl3): l 0.88 (9 H, t, J 6.6–7.3 Hz), 1.25–
1.49 (60 H, m), 1.66–1.71 (2 H, m), 1.96–2.05
(2 H, m), 3.23–3.38 (3 H, m), 3.66–3.77 (3 H,

276
Y. Watanabe et al. / Carbohydrate Research 332 (2001) 257–277
Determination of biological acti6ity.—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.
Cell culture.—Human monoblastic U937
cells were maintained in RPMI-1640 medium
supplemented with 10% FBS, 100 units/mL of
penicillin and 100 mg/mL of streptomycin
(growth medium).
m), 4.32 (1 H, m), 4.49 (1 H, ddd, J 3.7, 10.3,
46.1 Hz), 4.56–4.62 (2 H, m), 4.64 (1 H, ddd,
J 6.6, 10.3, 46.1 Hz), 7.19–7.37 (11 H, m,
containing NH). FABMS, (positive-ion): m/z
1090 (M+Na)+, 1068 (M+H)+. HR-
FABMS (positive-ion): Calcd for C59H97F3-
NNaO10P: 1090.6700; Found: 1090.6688.
2,6 - Anhydro - 3,7 - dideoxy - 4 - O - [(R) - 3-
(dodecyloxy)tetradecyl] - 7 - fluoro - 3 - [(R) - 3-
hydroxytetradecanamido] - 5 - O - phosphono -
glycero- -ido-heptonic acid (23a).—Com-
D
-
D
pound of 22a (10.3 mg, 0.010 mmol) was
treated as described in the formation of 17aa
from 16aa to give 23a (6.1 mg, 69%) as a
white powder; mp 112.5–113.5 °C. [h]2D5 +8.1°
(c 0.10, CHCl3). IR wmax(KBr): 3311, 2956,
1
2923, 2853, 1730, 1646 cm−1. H NMR (400
Production of TNFh by U937 cells.—U937
cells (1×104/200 mL/well) were plated in 96-
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 the data are
reproducible.
MHz, 5:1 CD3OD–CDCl3): l 0.89 (9 H, t, J
6.6 Hz), 1.29–1.56 (60 H, m), 1.76–1.77 (2 H,
m), 2.32 (1 H, dd, J 8.1, 14.6 Hz), 2.39 (1 H,
dd, J 3.7, 14.6 Hz), 3.43–3.48 (3 H, m),
3.76–3.80 (4 H, m), 3.91 (1 H, m), 4.20–4.73
(5 H, m, containing 1 H, d, J 3.7 Hz, at 4.60).
FABMS, (positive-ion): m/z 918 (M+Na)+.
FABMS, (negative-ion): m/z 894 (M−H)−.
HRFABMS
(positive-ion):
Calcd
for
C47H91FNNaO11P:
918.6232.
918.6211; Found:
2,6-Anhydro-3,7-dideoxy-3-(2,2-difluorotet-
radecanamido)-4-O-[(R)-3-(dodecyloxy)tetra-
decyl]-7-fluoro-5-O-phosphono-
D
-glycero- -
D
ido-heptonic acid (23b).—Compound 22b
(21.1 mg, 0.020 mmol) was treated as de-
scribed in the formation of 17aa from 16aa to
give 23b (16.7 mg, 93%) as a white powder;
mp 72.0–74.0 °C. [h]2D6 +3.7° (c 0.44, CHCl3).
IR wmax(KBr): 3327, 2922, 2852, 1691 cm−1
.
1H NMR (400 MHz, 9:1 CD3OD–CDCl3): l
0.90 (9 H, t, J 6.6–7.3 Hz), 1.29–1.57 (60 H,
m), 1.74–1.79 (2 H, m), 1.99–2.07 (2 H, m),
3.41–3.47 (3 H, m), 3.73–3.83 (3 H, m), 4.23–
4.34 (2 H, m), 4.41 (1 H, dd, J 3.7, 6.6 Hz),
4.60 (1 H, ddd, J 3.7, 10.3, 48.3 Hz), 4.63 (1
H, d, J 3.7 Hz), 4.77 (1 H, ddd, J 6.6, 10.3,
48.3 Hz). FABMS, (positive-ion): m/z 938
(M+Na)+, 916 (M+H)+. HRFABMS (pos-
itive-ion): Calcd for C47H89F3NNaO10P:
938.6074; Found: 938.6078. Anal. Calcd for
C47H89F3NO10P (916.2): C, 61.62; H, 9.79; F,
6.22; N, 1.53; P, 3.38. Found: C, 60.94; H,
9.94; F, 5.82; N, 1.50; P, 3.56.
Acknowledgements
This study was supported, in part, by the
Social Insurance Agency Fund commissioned
by the Japan Health Sciences Foundation.
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