Practical stereoselective synthesis of α-linked C-glucosamine propionic acid esters: Conversion to GLA-60 derivatives

Article abstract of DOI:10.1016/S0008-6215(01)00290-7

Radical C-glycosylation of glucosamine derivatives by acrylic acid esters gave the corresponding 3-(α-C-glucosyl)-propionate derivatives in moderate yields. One of them was used as a versatile synthon for GLA-60 derivatives. However, the biological activi

Full text of DOI:10.1016/S0008-6215(01)00290-7

Carbohydrate Research 337 (2002) 97–104
www.elsevier.com/locate/carres
Practical stereoselective synthesis of a-linked C-glucosamine
propionic acid esters: conversion to GLA-60 derivatives
Takanori Wakabayashi, Masao Shiozaki,* Shin-ichi Kurakata
Exploratory Chemistry Research Laboratories, Sankyo Co. Ltd., Hiromachi 1-2-58, Shinagawa-ku, Tokyo 140-8710, Japan
Received 7 August 2001; accepted 27 September 2001
Abstract
Radical C-glycosylation of glucosamine derivatives by acrylic acid esters gave the corresponding 3-(a-C-glucosyl)-propionate
derivatives in moderate yields. One of them was used as a versatile synthon for GLA-60 derivatives. However, the biological
activity of these compounds as LPS-antagonists was disappointing. © 2002 Elsevier Science Ltd. All rights reserved.
Keywords: GLA-60 derivatives; a-Linked C-glycosylation of glucosamine; LPS-antagonist
reactivity of a-oriented anomeric radical on the glucose
ring.2c Actually, the radical a-C-glucosylation reaction
of 2,3,4,6-tetra-O-acetyl-a-D-glucopyranosyl bromide
1. Introduction
with Bu3SnH and methyl acrylate using light or V-704
as a radical starter gave a corresponding a-linked C-
glucopyranoside in moderate yield.2l In the case of
N-acylglucosamines, this radical a-C-glucosylation was
carried out using allyltributyltin, vinyl phosphonate, or
acrylphosphonate to yield corresponding a-linked C-
glycosidic adducts in moderate yield.2j,2m,2o,2p However,
in these attempts, the amide groups such as the acetyl-
amino group derived from the starting glucosamines
were too stable to further modifications and functional-
izations for the access to the proposed compounds. One
of the serious problems is stable intramolecular d-lac-
tam formation. This prevents further conversion of
In our investigation of LPS-antagonists, the a
anomer A (a GLA-60 derivative) showed LPS-antago-
nist activity toward human monoblastic cells in spite of
its b anomer lacking in this activity.1 This result made
us interested in the biological activity of methylene
derivatives of A such as 15 (Fig. 1).
Syntheses of a-C-glycopyranosides have been studied
extensively, and many methods to control the stereo-
chemistry at the anomeric position have been re-
ported.2,3 Among these studies, we thought that the
radical a-C-glucosylation method should be one of
choice for our purposes, because of the stability and
3-(a-C-glucosyl)-propionate
Therefore, we also attempted another radical a-C-glu-
cosylation reaction of -glucosamine analogues to solve
derivatives
produced.
D
these restrictions. Herein we describe the results that
were applied for the further conversion to a-linked
C-glucosamine propionic acid derivatives of GLA-60 to
give an example.
Fig. 1. Structures of compound A and GLA-60.
2. Results and discussion
* Corresponding author. Tel.: +81-3-3492-3131; fax: +81-
3-54368570.
E-mail address: shioza@shina.sankyo.co.jp (M. Shiozaki).
As substrates, we used three kinds of glucosamine
analogues, that is, 2-deoxy-2-trifluoroacetamido-, 2-
0008-6215/02/$ - see front matter © 2002 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(01)00290-7

98
T. Wakabayashi et al. / Carbohydrate Research 337 (2002) 97–104
Table 1
Anomeric a-C-propionylation of 3,4,6-tri-O-acetyl-2-amino-2-deoxy-a-D-glucopyranosyl bromide analogues (1) with acrylic acid
esters
Entry
Substrate
R1
R2
Product
Yield (%)
1
2
3
4
1a
1a
1b
1c
NHCOCF3
NHCOCF3
NHCOOBn
NꢀCHꢁPMP
Bn
But
But
Bn
2a
2b
2c
2d
84
58
60
50
V-70=2,2%-azobis(2,4-dimethyl-4-methoxyvaleronitrile); PMP=4-methoxyphenyl.
deprotection of the 9-benzylcarbonate by chemoselec-
tive hydrogenolysis using the ethylenediamine-deacti-
vated Pd (Pd–C(en))5 as a catalyst gave the 9-OH
phosphonate 9 in 76% yield in four steps from 5.
Treatment of 9 with Me3O·BF4 and 2,6-di-tert-butyl-4-
methylpyridine (DTBMP) at room temperature gave
9-OMe 10 in 92% yield. On the other hand, treatment
of 9 with diethylaminosulfur trifluoride (DAST) gave
11 in 71% accompanying with a small amount of an
intramolecular cyclic 7,9-O-phenylphosphate. Finally,
the protective groups were removed from these com-
pounds to afford the objective compounds. Treatment
of compounds 8, 10, and 11 with CF3COOH in
CH2Cl2, and successive treatment of the resulted com-
pounds with hydrogen using Pd(OH)2-on-carbon as a
catalyst gave 12, 13, and 14, respectively, and these
compounds were further hydrogenolyzed using PtO2 as
a catalyst to afford 15, 16, and 17, respectively (Scheme
1).
Biological acti6ity.—The biological activity of com-
pounds 15, 16, and 17 on LPS-induced TNFa produc-
tion was investigated in vitro using human monoblastic
U937 cells. Compound 15 showed weak LPS-antagonis-
tic activity (IC50 50 nM), and compound 17 even
showed faint LPS-agonistic activity at a concentration
of 50 mM. But both compounds 16 and 17 were almost
inactive.
(benzyloxycarbonyl)amino-2-deoxy-, and 2-deoxy-2-(4-
methoxybenzyl)imino-a- -glucopyranosyl bromides
D
(1a, 1b, and 1c, respectively), which were anticipated to
be possible for further conversion. According to the
reported procedure of Kita et al.2l treatment of each
compound 1a,b,c with benzyl or tert-butyl acrylate in
the presence of tributyltin hydride using 2,2%-azobis-
(2,4-dimethyl-4-methoxyvaleronitrile) (V-70) as a radi-
cal initiator gave the corresponding a-linked 5-amino
C-glucosides 2a,b,c,d, without detection of b-linked
epimers (Table 1). In this series, the yield of trifluoroac-
etamido-benzyl ester 2a was 84%, and that of 4-
methoxybenzylimino benzyl ester 2d was 50%. This
reaction generally proceeds in moderate yield and thus
may have application for other 5-N substitution groups
in general.
Next, we used compound 2c for the conversion of 2
to propionic acid analogues of GLA-60 (15–17) ac-
cording to the modified procedure of the reported
method.1 Deprotection of the benzyloxycarbonyl group
from the C-5 carbamate of 2c by hydrogenolysis using
Pd-on-carbon as a catalyst, and successive amide for-
mation with (R)-3-benzyloxytetradecanoic acid using
3-(3-dimethylaminopropyl)-1-ethyl-carbodiimide hydro-
chloride (WSCDI) as a condensing agent, gave the C-5
amide which was treated with NaOMe in MeOH to
give the C-6,-7,-9-triol, and successive treatment of the
triol with 2,2-dimethoxypropane using catalytic amount
of TsOH as a catalyst gave the 7,9-O-isopropylidene
compound 4. The 6-OH group of 4 was esterified with
(R)-3-(tetradecanoyloxy)tetradecanoic
acid
using
3. Conclusions
DCC–DMAP as condensing reagents to give the C-6
ester 5 in 64% yield in five steps from 2c. Deblocking of
the 7,9-O-isopropylidene group of 5 in MeOH solution
using TsOH to give a diol 6, protection of the 9-OH
group of 6 with benzyl chloroformate using pyridine as
a base gave 7, diphenylphosphonate ester formation of
the 7-OH group of 7 with diphenyl chlorophosphate
using DMAP as a base gave 8, and regioselective
Thus, in this study we were able to explore a practi-
cal stereoselective synthesis for 5-amino-4,8-anhy-
dro-2,3,5-trideoxy-D-glycero-D-ido-nononic acid equiv-
alents, and we were able to apply this result to the
synthesis of GLA-60 derivatives. However, the biologi-
cal activity of these compounds as LPS-antagonists was
disappointing.

T. Wakabayashi et al. / Carbohydrate Research 337 (2002) 97–104
99
Scheme 1. Reagents and conditions: (a) (1) H2, Pd(OH)2–C, EtOH, rt, 20 min; (2) (R)-3-benzyloxytetradecanoic acid, WSCDI,
rt, 16 h, two steps 83%; (b) (1) NaOMe, MeOH, rt, 20 min; (2) 2,2-dimethoxypropane, TsOH·H2O, DMF, rt, 1 h, two steps 88%;
(c) (R)-3-(tetradecanoyloxy)tetradecanoic acid, DCC, DMAP, 88%; (d) TsOH, MeOH, rt, 2 h, 95%; (e) BnOOCCl, pyridine,
CH2Cl2, rt, 5 h, 93%; (f) diphenyl chlorophosphate, DMAP, CH2C12, rt, 4 h, 94%; (g) H2, Pd–C(en), THF, rt, 92%; (h)
Me3O·BF4, DTBMP, CH2Cl2, rt, 5 h, 92%; (i) DAST, CH2Cl2, 0 °C–rt, 16 h, 71%; (j) CF3COOH, CH2Cl2, rt, 4 h, then H2,
Pd(OH)2–C, EtOH, rt, 2 h, 12 in 73% (from 8), 13 in 76% (from 10), and 14 in 56% (from 11), respectively; (k) H2, PtO2, THF,
rt, 16 h, 93% (15), quant (16), and 94% (17), respectively.
4. Experimental
solution of tributyltinhydride (151 mg, 0.52 mmol) in
Et2O (4 mL) was slowly added at rt with stirring. After
1 h, the reaction mixture was filtered, concentrated in
vacuo, and chromatographed on a silica gel column.
Elution with 1:9, then 1:1 EtOAc–hexane gave 2a (192
mg, 84% yield). Rf 0.20 (3:7 EtOAc–hexane). IR
wmax(CHCl3) 3425, 2960, 1738, 1542, 1371, 1249, 1171,
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 polarimeter. 1H NMR spectra were recorded
with a JEOL-GSX 400 spectrometer using TMS as the
internal standard, and 13C NMR spectra were recorded
at 100 MHz using TMS as the internal standard. IR
absorption spectra were determined with an IR A-2
spectrophotometer, and mass spectra were obtained
with a JMS-700 mass spectrometer. Elemental analyses
were performed by the Institute of Science and Tech-
nology, Inc. Separation of the compounds by column
chromatography was done with Silica Gel 60 (230–400
mesh ASTM, E. Merck) under a slightly elevated pres-
sure (1.1–1.5 atm) for easy elution, and the quantity of
silica gel used was 50–100 times the weight charged on
the column. THF was distilled in the presence of radical
anions generated by sodium–benzophenone ketyl.
1
1046 cm−1. H NMR (CDCl3): l 7.39–7.31 (m, 5 H),
7.00 (d, 1 H, J 8.7 Hz, NH, D2O exchanged), 5.14, 5.12
(AB-q, 2 H, J 12.4 Hz), 5.04 (t, 1 H, J 5.5 Hz), 4.91 (t,
1 H, J 4.9 Hz), 4.45 (dd, 1 H, J 12.1 and 7.3 Hz, C9-H),
4.23–4.16 (m, 2 H, C5-H and C4-H or C8-H), 4.09 (dd,
1 H, J 12.1 and 4.5 Hz, C9-H), 3.99 (m, 1 H, C4-H or
C8-H), 2.48–2.43 (m, 2 H), 2.11 (s, 3 H), 2.09 (s, 3 H),
2.07 (s, 3 H), 1.93 (m, 1 H), 1.79 (m, 1 H). FABMS
(positive-ion): m/z 548 [M+H]+, 570 [M+Na]+. HR-
FABMS (positive-ion): Calcd for C24H29F3NO10:
548.1744. Found: 548.1738. Anal. Calcd for
C24H28F3NO10 (547.5): C, 52.65; H, 5.15; N, 2.56; F,
10.41. Found: C, 52.54; H, 5.10; N, 2.43; F, 10.32.
6,7,9-Tri-O-acetyl-4,8-anhydro-2,3,5-trideoxy-5-trifl-
,
DMF and pyridine were dried by storage over 4 A
molecular sieves.
uoroacetylamino-D-glycero-D-ido-nononic acid tert-butyl
6,7,9-Tri-O-acetyl-4,8-anhydro-2,3,5-trideoxy-5-trifl-
ester (2b).—Compound 1b (237 mg, 0.51 mmol) was
treated with tert-butyl acrylate as described in the
formation of 2a from 1a and benzyl acrylate to give 2b
(152 mg, 58% yield) as a gum. Rf 0.35 (7:3 Et2O–hex-
ane). IR: wmax(film) 3317, 2980, 1747, 1728, 1554, 1370,
uoroacetylamino-
ester (2a).—To a suspension of 2-acetamido-3,4,6-tri-
O-acetyl-2-deoxy-a- -glucopyranosyl bromide (1a)
D-glycero-D-ido-nononic acid benzyl
D
(194 mg, 0.42 mmol), 2,2%-azobis-(2,4-dimethyl-4-
methoxyvaleronitrile) (V-70) (160 mg, 0.52 mmol) and
benzyl acrylate (680 mg, 4.2 mmol) in Et2O (4 mL), a
1
1228, 1157, 1094, 1039, 758 cm−1. H NMR (CDCl3):
l 6.98 (d, 1 H, J 8.7 Hz, NH, D2O exchanged), 5.05 (t,

100
T. Wakabayashi et al. / Carbohydrate Research 337 (2002) 97–104
1 H, J 5.9 Hz), 4.94 (t, 1 H, J 5.1 Hz), 4.44 (dd, 1 H,
J 11.9 and 7.1 Hz, C9-H), 4.22 (m, 1 H, C5-H, changed
to dd, J 3.5, 6.4 Hz, on addition of D2O), 4.20–4.14 (m,
2 H, C9-H and C4-H or C8-H), 4.00 (m, 1 H, C4-H or
C8-H), 2.36–2.31 (m, 2 H), 2.12 (s, 3 H), 2.11 (s, 3 H),
2.09 (s, 3 H), 1.88 (m, 1 H), 1.72 (m, 1 H), 1.45 (s, 9 H).
13C NMR (100 MHz, CDCl3): l 171.9, 170.5, 170.1,
168.7, 156.9, 115.7, 80.8, 71.9, 68.6, 66.8, 60.7, 50.1,
30.9, 28.0, 23.7, 20.7, 20.6. FABMS (positive-ion): m/z
514 [M+H]+, 512 [M−H]+, 536 [M+Na]+, 552
[M+K]+ (on addition of KI), 590 [M+2K−H]+.
HRFABMS (positive-ion): Calcd for C21H30F3KNO10:
552.1459. Found: 552.1477.
Pd(OH)2–C (115 mg) was stirred under a hydrogen
atmosphere at rt for 20 min. The reaction mixture was
filtered and concentrated in vacuo. The crude product
was dissolved in CH2Cl2 (10 mL) then (R)-3-(benzy-
loxy)tetradecanoic acid (381 mg, 1.14 mmol) and WSCI
(1.16 mmol) were added. After stirring overnight at rt,
the reaction mixture was diluted with EtOAc, washed
with water and brine, dried over Na2SO4 and filtered.
The filtrate was concentrated in vacuo and chro-
matographed on a silica gel column. Elution with 3:7,
then 1:1 EtOAc–hexane gave 3 (626 mg, 83% yield). Rf
0.43 (2:3 EtOAc–hexane). IR wmax(CHCl3) 3429, 3356,
2928, 2856, 1743, 1671, 1514, 1456, 1369, 1251, 1154,
1
6,7,9-Tri-O-acetyl-4,8-anhydro-2,3,5-trideoxy-5-ben-
1045 cm−1. H NMR (CDCl3): l 7.34–7.28 (m, 5 H),
zyloxycarbonylamino-
D
-glycero-
D
-ido-nononic acid tert-
6.64 (d, 1 H, J 8.8 Hz), 5.02 (dd, 1 H, J 7.8 and 6.9 Hz),
4.89 (t, 1 H, J 6.6 Hz), 4.56, 4.52 (ABq, 2 H, J 11.3
Hz), 4.35–4.28 (m, 2 H), 4.12–4.06 (m, 2 H), 3.85 (m,
1 H), 3.79 (m, 1 H), 2.46 (dd, 1 H, J 15.1 and 3.8 Hz),
2.36 (dd, 1 H, J 15.0 and 7.0 Hz), 2.28–2.14 (m, 2 H),
2.10 (s, 3 H), 2.05 (s, 3 H), 1.89 (s, 3 H), 1.87–1.50 (m,
4 H), 1.44 (s, 9 H), 1.32–1.25 (m, 18 H), 0.88 (t, 3 H,
J 6.8 Hz). 13C NMR (100 MHz, CDCl3): l 172.1, 171.1,
170.6, 170.3, 169.3, 138.0, 128.4, 128.1, 127.8, 80.5,
76.2, 71.3, 70.8, 70.5, 69.8, 67.7, 61.5, 49.8, 41.2, 33.6,
31.9, 31.0, 29.61, 29.55, 29.3, 28.1, 25.2, 22.7, 22.4, 20.8,
20.7, 20.5, 14.1. FABMS (positive-ion): m/z 734 [M+
H]+, 756 [M+Na]+. HRFABMS (positive-ion): Calcd
for C40H64NO11: 734.4479. Found: 734.4498.
butyl ester (2c).—Compound 1b (1.885 g, 3.75 mmol)
was treated with tert-butyl acrylate as described in the
formation of 2a from 1a and benzyl acrylate to gave 15
(1.124 g, 54% yield). Rf 0.38 (2:3 EtOAc–hexane). IR
wmax(KBr) 3351, 2977, 1746, 1729, 1533, 1330, 1237,
1
1154, 1039 cm−1. H NMR (CDCl3): l 7.36–7.31 (m, 5
H), 5.15–5.05 (m, 4 H), 4.95 (t, 1 H, J 7.0 Hz), 4.32
(dd, 1 H, J 12.0 and 6.0 Hz), 4.13–4.00 (m, 3 H), 3.87
(m, 1 H), 2.38–2.25 (m, 2 H), 2.10 (s, 3 H), 2.02 (s, 3
H), 2.01 (s, 3 H), 1.94 (m, 1 H), 1.80 (m, 1 H), 1.45 (s,
9 H). 13C NMR (100 MHz, CDCl3): l 172.1, 170.6,
169.1, 155.7, 136.2, 128.5, 128.2, 128.1, 80.6, 71.4, 70.4,
69.9, 67.8, 67.0, 61.6, 52.2, 31.1, 28.0, 22.1, 20.7.
FABMS (positive-ion): m/z 552 [M+H]+, 574 [M+
4,8 - Anhydro - 5 - [(R) - 3 - (benzyloxy)tetradecanoyl -
Na]+.
HRFABMS
(positive-ion):
Calcd
for
amino]-2,3,5-trideoxy-7,9-O-isopropylidene-
D-glycero-
C27H38NO11: 552,2445. Found: 552.2435. Anal. Calcd
for C27H37NO11: C, 58.79; H, 6.76; N, 2.54. Found: C,
58.66; H, 6.66; N, 2.48.
D
-ido-nononic acid tert-butyl ester (4).—A solution of 3
(626 mg, 0.853 mmol) in MeOH (10 mL) was treated
with 1 M NaOMe in MeOH (0.1 mL) for 20 min at rt,
then evaporated in vacuo. To the residue were added
DMF (10 mL), 2,2-dimethoxypropane (5 mL) and
TsOH·H2O (40 mg, 0.21 mmol). After stirring for 1 h at
rt, the reaction mixture was quenched with satd aq
NaHCO3, extracted with EtOAc, washed with water
and brine, dried over Na2SO4 and filtered. The filtrate
was concentrated in vacuo and chromatographed on a
silica gel column. Elution with 1:1, then 4:1 EtOAc–
hexane gave 4 (485 mg, 88% yield). Rf 0.34 (2:3
EtOAc–hexane). IR wmax(KBr) 3440, 3315, 2925, 2854,
6,7,9-Tri-O-acetyl-4,8-anhydro-2,3,5-trideoxy-5-[(4-
methoxybenzylidene)amino] -
acid benzyl ester (2d).—3,4,6-Tri-O-acetyl-2-deoxy-2-
[(4 - methoxybenzylidene)amino] - a - - glucopyranosyl
D - glycero - D - ido - nononic
D
bromide (1c) (1.885 g, 3.75 mmol) was treated as de-
scribed in the formation of 2a from 1a to give 2d (1.124
g, 54%). Rf 0.38 (2:3 EtOAc–hexane). IR wmax(film)
3030-2840, 1745, 1641, 1605, 1577, 1513 cm−1 1H
.
NMR (CDCl3): l 8.17 (s, 1 H), 7.66 (d, 2 H, J 8.8 Hz),
7.34 (s, 5 H), 6.90 (d, 2 H, J 8.8 Hz), 5.47 (dd, 1 H, J
10.2, 9.5 Hz, C6-H), 5.12 (s, 2 H), 5.04 (t, 1 H, J 9.5
Hz), 4.29 (dd, 1 H, J 11.7, 4.4 Hz), 4.04–3.96 (m, 3 H,
C4,6,9-H), 3.84 (s, 3 H), 3.66 (dd, 1 H, 9.5, 5.9 Hz,
C8-H), 2.51–2.42 (m, 2 H), 2.37–2.31 (m, 2 H), 2.08 (s,
3 H), 2.03 (s, 3 H), 1.86 (s, 3 H). FABMS (positive-ion):
m/z 570 [M+H]+, [M+Na]+. HRFABMS (positive-
ion): Calcd for C30H36NO10: 570.2339. Found:
570.2337. Anal. Calcd for C30H35NO10: C, 63.26; H,
6.19; N, 2.46. Found: C, 63.16; H, 6.33; N, 2.55.
6,7,9-Tri-O-acetyl-4,8-anhydro-5-[(R)-3-(benzyloxy-
1728, 1630, 1456, 1368, 1156, 1100 cm−1. H NMR
1
(CDCl3): l 7.39–7.28 (m, 5 H), 6.50 (d, 1 H, J 7.2 Hz),
4.63, 4.51 (ABq, 2 H, J 11.3 Hz), 4.16 (m, 1 H), 4.08
(m, 1 H), 3.84–3.77 (m, 2 H), 3.68 (t, 1 H, J 10.4 Hz),
3.55–3.45 (m, 2 H), 3.32 (m, 1 H), 2.60 (d, 1 H, J 1.9
Hz), 2.53 (dd, 1 H, J 15.0 and 3.8 Hz), 2.42 (dd, 1 H,
J 15.0 and 6.6 Hz), 2.22 (m, 1 H), 2.10 (m, 1 H),
1.84–1.52 (m, 4 H), 1.49 (s, 3 H), 1.44 (s, 9 H), 1.42 (s,
3 H), 1.39–1.25 (m, 18 H), 0.88 (t, 3 H, J 6.9 Hz). 13C
NMR (100 MHz, CDCl3): l 172.3, 138.1, 128.5, 127.9,
127.8, 99.8, 80.5, 76.7, 75.4, 74.2, 71.3, 69.9, 64.3, 62.6,
54.0, 40.9, 33.4, 31.9, 31.2, 29.61, 29.55, 29.3, 29.1, 25.2,
22.7, 20.6, 19.1, 14.1. FABMS (positive-ion): m/z 648
tetradecanoyl)amino]-2,3,5-trideoxy-D -glycero-D -ido-
nononic acid tert-butyl ester (3).—A solution of 2c (570
mg, 1.03 mmol) in EtOH (5 mL) containing 20%

T. Wakabayashi et al. / Carbohydrate Research 337 (2002) 97–104
101
[M+H]+. HRFABMS (positive-ion): Calcd for
C37H62NO8: 648.4476. Found: 648.4470.
4,8 - Anhydro - 9 - O - benzyloxycarbonyl - 5 - [(R) - 3-
(benzyloxy)tetradecanoylamino] - 2,3,5 - trideoxy - 6 - O-
4,8 - Anhydro - 5 - [(R) - 3 - (benzyloxy)tetradecanoyl-
amino]-2,3,5-trideoxy-7,9-O-isopropylidene-6-O-[(R)-
[(R) - 3 - (tetradecanoyloxy)tetradecanoyl] - D - glycero - D-
ido-nononic acid tert-butyl ester (7).—To a solution of 6
(644 mg, 0.616 mmol) and pyridine (1 mL) in CH2Cl2
(12 mL) were gradually added ClCOOBn (1 mL) with
stirring at rt. After 5 h, the reaction mixture was
concentrated in vacuo, and satd aq NaHCO3 was
added. The aqueous layer was extracted with EtOAc,
and washed with water and brine, and the organic
extract was dried over Na2SO4 and filtered. The filtrate
was concentrated in vacuo and chromatographed on a
silica gel column. Elution with (1:4, then 3:7) EtOAc–
hexane gave 7 (676 mg, 93% yield). Rf 0.40 (3:7
EtOAc–hexane). IR wmax(KBr) 3484, 3297, 2921, 2852,
1730, 1650, 1292 cm−1. 1H NMR (CDCl3): l 7.40–7.27
(m, 5 H), 6.49 (d, 1 H, J 7.3 Hz), 5.17 (s, 2 H), 5.09 (m,
1 H), 4.90 (dd, 1 H, J 10.6 and 8.4 Hz), 4.55, 4.51
(ABq, 2 H, J 11.7 Hz), 4.42–4.36 (m, 2 H), 4.24 (m, 1
H), 4.12 (m, 1 H), 3.81 (m, 1 H), 3.62 (m, 1 H), 3.55 (m,
1 H), 3.27 (d, 1 H, J 3.7 Hz) 2.49 (d, 2 H, J 5.9 Hz),
2.37–2.20 (m, 5 H), 2.07 (m, 1 H), 1.83 (m, 1 H),
1.63–1.53 (m, 5 H), 1.42 (s, 9 H), 1.36–1.25 (m, 58 H),
0.88 (t, 9 H, J 6.6 Hz). FABMS (positive-ion): m/z 1178
[M+H]+, 1216 [M+K]+ (on addition of KI). HR-
FABMS (positive-ion): Calcd for C70H115NKO13:
1216.8006. Found: 1216.8030.
3 - (tetradecanoyloxy)tetradecanoyl] -
D - glycero - D - ido-
nononic acid tert-butyl ester (5).—To a solution of 4
(403 mg, 0.622 mmol), (R)-3-(tetradecanoyloxy)-
tetradecanoic acid (377 mg, 0.83 mmol) and DMAP
(10.6 mg) in CH2Cl2 were added DCC (160 mg, 0.776
mmol) at rt. After stirring for 7 h, the reaction mixture
was filtered, concentrated in vacuo and chro-
matographed on a silica gel column. Elution with 3:17,
then 1:3 EtOAc–hexane gave 5 (596 mg, 88% yield). Rf
0.39 (1:4 EtOAc–hexane). IR wmax(KBr) 3419, 2921,
2851, 1732, 1661, 1521, 1469, 1368, 1200, 1161, 1111
1
cm−1. H NMR (CDCl3): l 7.38–7.25 (m, 5 H), 6.34
(d, 1 H, J 7.5 Hz), 5.14 (m, 1 H), 5.01 (dd, 1 H, J 10.8
and 9.6 Hz), 4.56, 4.51 (ABq, 2 H, J 11.8 Hz), 4.29 (m,
1 H), 4.13 (m, 1 H), 3.84–3.78 (m, 2 H), 3.71–3.66 (m,
2 H), 3.42 (m, 1 H), 2.60 (dd, 1 H, J 15.1 and 6.9 Hz),
2.50 (dd, 1 H, J 14.9 and 5.9 Hz), 2.35–2.25 (m, 4 H),
2.18 (m, 1 H), 2.02 (m, 1 H), 1.88 (m, 1 H), 1.67–1.52
(m, 5 H), 1.46 (s, 3 H), 1.43 (s, 9 H), 1.36 (s, 3 H),
1.32–1.25 (m, 58 H), 0.88 (t, 9 H, J 6.7 Hz). 13C NMR
(100 MHz, CDCl3): l 173.3, 172.1, 171.3, 171.0, 138.5,
128.4, 127.64, 127.58, 99.7, 80.4, 76.1, 74.3, 72.8, 71.2,
70.2, 70.0, 64.7, 62.7, 52.8, 41.6, 39.3, 34.4, 33.9, 33.8,
31.9, 31.1, 29.7, 29.53, 29.46, 29.3, 29.2, 29.0, 28.1, 25.2,
25.0, 22.7, 20.3, 19.1, 14.1. FABMS (positive-ion): m/z
1084 [M+H]+. HRFABMS (positive-ion): Calcd for
C65H114NO11: 1084.8391. Found: 1084.8365.
4,8 - Anhydro - 9 - O - benzyloxycarbonyl - 5 - [(R) - 3-
(benzyloxy)tetradecanoylamino] - 2,3,5 - trideoxy - 7 - O-
diphenylphosphono-6-O-[(R)-3-(tetradecanoyloxy)tetra-
decanoyl]-D-glycero-D-ido-nononic acid tert-butyl ester
4,8 - Anhydro - 5 - [(R) - 3 - (benzyloxy)tetradecanoyl-
amino]-2,3,5-trideoxy-6-O-[(R)-3-(tetradecanoyloxy)-
tetradecanoyl]-D-glycero-D-ido-nononic acid tert-butyl
(8).—To a solution of 7 (676 mg, 0.574 mmol) and
DMAP (176 mg, 1.441 mmol) in CH2Cl2 (6 mL) were
added diphenyl chlorophosphate (311 mg, 1.157 mmol)
in CH2Cl2 (1 mL) with stirring at rt. After 4 h, the
reaction mixture was quenched with satd aq NaHCO3,
and extracted with EtOAc. The organic extract was
washed with water and brine, dried over Na2SO4, and
filtered. The filtrate was concentrated in vacuo and
chromatographed on a silica gel column. Elution with
1:4, then 3:7 EtOAc–hexane gave 8 (762 mg, 94%
yield). Rf 0.40 (1:3 EtOAc–hexane). IR wmax(KBr) 3335,
ester (6).—To a suspension of 5 (596 mg, 0.550 mmol)
in MeOH (20 mL) were added TsOH·H2O (10.5 mg) at
rt. After stirring for 2 h, the reaction mixture was
quenched with NEt3, evaporated and chromatographed
on a silica gel column. Elution with 2:3, then 1:1
EtOAc–hexane gave 6 (546 mg, 95% yield). Rf 0.50 (1:1
EtOAc–hexane). IR wmax(KBr) 3472, 3302, 2922, 2852,
1730, 1649, 1545, 1468, 1368, 1161, 1101 cm−1 1H
.
NMR (CDCl3): l 7.36–7.25 (m, 5 H), 6.55 (d, 1 H, J
7.5 Hz), 5.09 (m, 1 H), 4.91 (dd, 1 H, J 10.4 and 8.3
Hz), 4.56, 4.52 (ABq, 2 H, J 11.6 Hz), 4.20 (m, 1 H),
4.12 (m, 1 H), 3.84–3.78 (m, 2 H), 3.59 (m, 1 H), 3.51
(m, 1 H), 3.23 (d, 1 H, J 3.7 Hz) 2.50 (d, 2 H, J 5.8 Hz),
2.38–2.21 (m, 5 H), 2.11 (m, 1 H), 1.85 (m, 1 H),
1.68–1.53 (m, 5 H), 1.44 (s, 9 H), 1.32–1.26 (m, 58 H),
0.88 (t, 9 H, J 6.7 Hz). 13C NMR (100 MHz, CDCl3):
l 174.6, 172.4, 172.2, 171.3, 138.4, 128.4, 127.7, 80.6,
76.2, 74.3, 72.54, 72.47, 71.2, 71.1, 69.8, 62.4, 51.4, 41.4,
40.5, 34.8, 34.5, 34.0, 31.9, 31.2, 29.7, 29.5, 29.3, 29.1,
28.1, 25.3, 25.2, 24.9, 22.7, 20.7, 14.1. FABMS (posi-
tive-ion): m/z 1044 [M+H]+, 1066 [M+Na]+. HR-
FABMS (positive-ion): Calcd for C62H109NNaO11:
1066.7899. Found: 1066.7874.
2922, 2852, 1744, 1729, 1654, 1265, 954 cm−1 1H
.
NMR (CDCl3): l 7.37–7.12 (m, 20 H), 6.45 (d, 1 H, J
7.4 Hz), 5.23 (dd, 1 H, J 10.0 and 8.1 Hz), 5.14–5.07
(m, 3 H), 5.02 (ABq, 1 H, J 12.2 Hz), 4.65 (dd, 1 H, J
17.1 and 8.4 Hz), 4.56, 4.51 (ABq, 2 H, J 11.7 Hz),
4.30–4.16 (m, 4 H), 3.90–3.82 (m, 2 H), 2.51–2.28 (m,
4 H), 2.22–2.15 (m, 3 H), 2.06 (m, 1 H), 1.84 (m, 1 H),
1.67–1.37 (m, 14 H), 1.32–1.25 (m, 58 H), 0.88 (t, 9 H,
J 6.8 Hz). 13C NMR (100 MHz, CDCl3): l 173.4, 172.2,
171.4, 170.7, 154.7, 150.3, 138.5, 135.1, 129.8, 128.5,
128.4, 128.3, 128.2, 127.6, 127.5, 125.6, 120.1, 80.5,
76.2, 74.3, 72.3, 71.2, 70.1, 69.9, 69.8, 69.7, 65.3, 51.3,
41.7, 39.2, 34.3, 34.2, 34.1, 31.9, 31.0, 29.7, 29.5, 29.3,
29.2, 28.1, 25.2, 25.1, 25.0, 22.7, 20.7, 14.1. FABMS

102
T. Wakabayashi et al. / Carbohydrate Research 337 (2002) 97–104
(positive-ion): m/z 1410 [M+H]+, 1432 [M+Na]+.
HRFABMS (positive-ion): Calcd for C82H125NO16P:
1410.8736. Found: 1410.8744. Anal. Calcd for
C82H124NO16P: C, 69.81; H, 8.86; N, 0.99; P, 2.20.
Found: C, 69.64; H, 8.69; N, 0.96; P, 2.00.
4,8 - Anhydro - 5 - [(R) - 3 - (benzyloxy)tetradecanoyl-
amino] - 2,3,5 - trideoxy - 7 - O - diphenylphosphono - 6 - O-
[(R) - 3 - (tetradecanoyloxy)tetradecanoyl] - D - glycero - D-
4.29 (m, 1 H), 4.20 (m, 1 H), 3.85 (m, 1 H), 3.74 (m, 1
H), 3.44 (d, 2 H, J 3.5 Hz), 3.22 (s, 3 H), 2.49–2.16 (m,
7H), 2.08 (m, 1 H), 1.86 (m, 1 H), 1.68–1.40 (m, 14 H),
1.32–1.25 (m, 58 H), 0.88 (t, 9 H, J 6.8 Hz). 13C NMR
(100 MHz, CDCl3): l 173.4, 172.3, 171.3, 170.9, 150.4,
138.6, 129.8, 128.3, 127.7, 127.5, 125.6, 120.2, 80.4,
76.2, 74.7, 72.5, 71.2, 70.7, 70.2, 69.8, 59.2, 51.7, 41.8,
39.1, 34.3, 34.1, 31.9, 31.1, 29.7, 29.6, 29.4, 29.2, 28.1,
25.2, 25.1, 25.0, 22.7, 20.5, 14.1. FABMS (positive-ion):
m/z 1290 [M+H]+, 1312 [M+Na]+. HRFABMS
(positive-ion): Calcd for C75H121NO14P: 1290.8521.
Found: 1290.8525. Anal. Calcd for C75H120NO14P: C,
69.79; H, 9.37; N, 1.09; P, 2.40. Found: C, 69.37; H,
9.02; N, 1.12; P, 2.33.
ido-nononic acid tert-butyl ester (9).—A solution of 8
(305 mg, 0.216 mmol) in THF (2 mL) containing
Pd(en)–C (32 mg) was stirred under a hydrogen atmo-
sphere at rt for 1 h. The reaction mixture was filtered,
concentrated in vacuo and chromatographed on a silica
gel column. Elution with 3:7, then 1:1 EtOAc–hexane
gave 9 (254 mg, 92% yield). Rf 0.43 (2:3 EtOAc–hex-
ane). IR wmax(KBr) 2922, 2852, 1736, 1650, 1492, 1197,
4,8 - Anhydro - 5 - [(R) - 3 - (benzyloxy)tetradecanoyl-
amino] - 2,3,5,9 - tetradeoxy - 7 - O - diphenylphosphono - 9-
1160, 1046, 1026, 968 cm−1
.
1H NMR (CDCl3): l
fluoro-6-O-[(R)-3-(tetradecanoyloxy)tetradecanoyl]-
D-
7.37–7.16 (m, 15 H), 6.42 (d, 1 H, J 7.4 Hz), 5.25 (dd,
1 H, J 10.4 and 8.5 Hz), 5.11 (m, 1 H), 4.66 (dd, 1 H,
J 17.7 and 8.7 Hz), 4.57, 4.51 (ABq, 2 H, J 11.7 Hz),
4.30 (m, 1 H), 4.20 (m, 1 H), 3.85 (m, 1 H), 3.69–3.54
(m, 3 H), 3.08 (dd, 1 H, J 7.9 and 6.9 Hz), 2.44–2.03
(m, 8 H), 1.85 (m, 1 H), 1.66–1.21 (m, 72 H) 0.88 (t, 9
H, J 6.9 Hz). 13C NMR (100 MHz, CDCl3): l 173.4,
172.5, 171.4, 170.8, 150.3, 138.5, 129.9, 128.3, 127.6,
127.5, 125.8, 120.1, 80.6, 76.2, 74.3, 72.4, 71.9, 71.2,
70.4, 69.8, 60.4, 51.8, 41.7, 39.1, 34.4, 34.2, 34.0, 31.9,
31.3, 29.7, 29.6, 29.3, 29.2, 28.1, 25.2, 25.1, 25.0, 22.7,
20.5, 14.1. FABMS (positive-ion): m/z 1276 [M+H]+,
1298 [M+Na]+. HRFABMS (positive-ion): Calcd for
C74H118NNaO14P: 1298.8188. Found: 1298.8207. Anal.
Calcd for C74H118NO14P: C, 69.62; H, 9.32; N, 1.10; P,
2.43. Found: C, 69.55; H, 9.15; N, 1.16; P, 2.51.
glycero- -ido-nononic acid tert-butyl ester (11).—To a
D
solution of 9 (105 mg, 0.082 mmol) in CH2Cl2 (4 mL)
was gradually added DAST (200 mL), with stirring at
0 °C. After stirring overnight, the reaction mixture was
quenched with satd aq NaHCO3, and extracted with
EtOAc. The organic extract was washed with water and
brine, dried over Na2SO4 and filtered. The filtrate was
concentrated in vacuo and chromatographed on a silica
gel column. Elution with 1:4, then 1:1 EtOAc–hexane
gave 11 (74.7 mg, 71% yield). Rf 0.58 (3:7 EtOAc–hex-
ane). IR wmax(KBr) 3423, 3371, 2927, 2855, 1724, 1674,
1491, 1292, 1161, 963 cm−1 1H NMR (CDCl3): l
.
7.36–7.16 (m, 15 H), 6.44 (d, 1 H, J 7.2 Hz), 5.26 (dd,
1 H, J 10.2 and 8.4 Hz), 5.10 (m, 1 H), 4.63 (dd, 1 H,
J 17.6 and 8.8 Hz), 4.57, 4.51 (ABq, 2 H, J 11.7 Hz),
4.42 (m, 2 H, JHF 46.9 Hz), 4.31–4.19 (m, 2 H),
3.87–3.77 (m, 2 H), 2.50–2.29 (m, 4 H), 2.23–2.15 (m,
3 H), 2.08 (m, 1 H), 1.86 (m, 1 H), 1.67–1.42 (m, 14 H),
1.39–1.25 (m, 58 H), 0.88 (t, 9 H, J 6.8 Hz). 13C NMR
(100 MHz, CDCl3): l 173.4, 172.3, 171.5, 170.9, 150.3,
138.6, 129.9, 128.4, 127.6, 127.5, 125.7, 120.1, 80.9,
80.5, 76.2, 73.9, 72.8, 71.2, 70.6, 70.3, 69.9, 51.6, 41.7,
39.2, 34.3, 34.2, 34.1, 31.9, 31.1, 29.7, 29.6, 29.4, 29.2,
28.1, 25.2, 25.1, 25.0, 22.7, 20.2, 14.1. FABMS (posi-
tive-ion): m/z 1278 [M+H]+, 1300 [M+Na]+. HR-
FABMS (positive-ion): Calcd for C74H118FNO13P:
1278.8325. Found: 1278.8334. Anal. Calcd for
C74H117FNO13P: C, 69.51; H, 9.22; N, 1.10; F, 1.49; P,
2.42. Found: C, 69.19; H, 8.43; N, 1.03; F, 1.38; P, 2.47
4,8-Anhydro-2,3,5-trideoxy-7-O-diphenylphosphono-5-
[(R)-3-(hydroxy)tetradecanoylamino]-6-O-[(R)-3-(tetra-
4,8 - Anhydro - 5 - [(R) - 3 - (benzyloxy)tetradecanoyl-
amino] - 2,3,5 - trideoxy - 7 - O - diphenylphosphono - 9 - O-
methyl-6-O-[(R)-3-(tetradecanoyloxy)tetradecanoyl]-
D-
glycero- -ido-nononic acid tert-butyl ester (10).—A so-
D
lution of 8 (97.9 mg, 0.069 mmol) in THF (1 mL)
containing Pd(en)–C (9.8 mg) was stirred under a
hydrogen atmosphere at rt for 3 h. The reaction mix-
ture was filtered and concentrated in vacuo to give
crude 9 (88.1 mg, 99%). The crude 9 was dissolved in
CH2Cl2 (2 mL). 2,6-Di-tert-butyl-4-methylpyridine (59
mg) and Me3O·BF4 (39 mg) were added portionwise to
the solution. After stirring overnight at rt, the reaction
mixture was diluted with EtOAc, washed with satd aq
NaHCO3, water and brine, and the organic extract was
dried over Na2SO4 and filtered. The filtrate was concen-
trated in vacuo and chromatographed on a silica gel
column. Elution with 1:9, then 2:3 EtOAc–hexane gave
10 (76.3 mg, 85% yield). Rf 0.32 (3:7 EtOAc–hexane).
IR wmax(KBr) 3423, 3372, 2927, 2855, 1725, 1674, 1490,
decanoyloxy)tetradecanoyl] -
D - glycero - D - ido - nononic
acid (12).—A solution of 8 (71.5 mg, 0.051 mmol) in
CH2Cl2 (2 mL) was treated with CF3COOH (0.2 mL)
for 4 h at rt, then evaporated in vacuo to give nononic
acid, which was dissolved in EtOH (2 mL). Pd(OH)2–C
(21 mg) was added. The mixture was stirred 2 h under
a hydrogen atmosphere at rt and was filtered. The
filtrate was concentrated in vacuo and chro-
1
1290, 1161, 1087, 1025, 959 cm−1. H NMR (CDCl3):
l 7.35–7.17 (m, 15 H), 6.38 (d, 1 H, J 7.4 Hz), 5.24 (dd,
1 H, J 10.3 and 8.4 Hz), 5.11 (m, 1 H), 4.70 (dd, 1 H,
J 17.4 and 8.7 Hz), 4.56, 4.51 (ABq, 2 H, J 11.7 Hz),

T. Wakabayashi et al. / Carbohydrate Research 337 (2002) 97–104
103
matographed on a silica gel column. The crude product
was purified by preparative TLC to give 12 (42.1 mg,
73% yield). Rf 0.60 (1:9 MeOH–CH2Cl2). IR wmax(KBr)
3414, 2924, 2854, 1738, 1648, 1664, 1490, 1457, 1275,
3.95 (m, 1 H), 3.83 (m, 1 H, J 9.0, JFH 23.3 Hz)
2.51–2.21 (m, 8 H), 2.01–1.91 (m, 2 H), 1.58–1.25 (m,
62 H) 0.88 (t, 9 H, J 6.7 Hz). 13C NMR (100 MHz,
CD3OD): l 176.1, 174.1, 173.1, 171.1, 150.3, 129.9,
125.7, 120.1, 80.9, 74.0, 72.6, 70.6, 70.4, 70.1, 68.7, 51.6,
42.8, 39.5, 37.2, 34.4, 34.3, 31.9, 29.7, 29.6, 29.5, 29.4,
29.2, 25.5, 25.1, 25.0, 22.7, 20.2, 14.1. FABMS (posi-
tive-ion): m/z 1132 [M+H]+, 1154 [M+Na]+. HR-
FABMS (positive-ion): Calcd for C63H104FNO13P:
1132.7229. Found: 1132.7230. Anal. Calcd for
C63H103FNO13P: C, 66.82; H, 9.17; F, 1.68; N, 1.24; P,
2.74. Found: C, 66.63; H, 9.01; F, 1.70; N, 1.17; P, 2.62.
4,8-Anhydro-2,3,5-trideoxy-5-[(R)-3-(hydroxy)tetra-
decanoylamino]-7-O-phosphono-6-O-[(R)-3-(tetrade-
1
1189, 1025, 964 cm−1. H NMR (CD3OD): l 7.41-7.19
(m, 10 H), 5.42 (dd, 1 H, J 10.4 and 8.7 Hz), 5.14 (m,
1 H), 4.68 (dd, 1 H, J 17.7 and 8.9 Hz), 4.33 (dd, 1 H,
J 10.6 and 5.7 Hz), 4.08 (m, 1 H), 3.93 (m, 1 H), 3.75
(m, 1 H), 3.68 (dd, 1 H, J 12.0 and 1.6 Hz), 3.59 (dd, 1
H, J 12.2 and 4.9 Hz), 2.55–2.23 (m, 6 H), 2.19 (t, 2 H,
J 7.3 Hz), 2.10 (m, 1 H), 1.90 (m, 1 H), 1.59–1.28 (m,
62 H) 0.89 (t, 9 H, J 6.6 Hz). 13C NMR (100 MHz,
CD3OD): l 173.4, 173.0, 170.2, 150.3, 129.6, 125.5,
119.8, 75.5, 72.7, 71.5, 70.7, 69.5, 68.2, 60.2, 51.2, 43.3,
38.4, 37.1, 33.8, 33.6, 31.6, 29.4, 29.3, 29.0, 28.8, 28.7,
25.3, 24.7, 22.3, 20.6, 13.0. FABMS (positive-ion): m/z
1131 [M+H]+, 1153 [M+Na]+, 1169 [M+K]+ (on
addition of KI), 1207 [M+2K−H]+. HRFABMS
(positive-ion): Calcd for C63H104NNaO14P: 1152.7092.
Found: 1152.7097. Anal. Calcd for C63H104NO14P: C,
66.94; H, 9.27; N, 1.24; P, 2.74. Found: C, 66.40; H,
8.79; N, 1.17; P, 2.95.
canoyloxy)tetradecanoyl]-D-glycero-D-ido-nononic acid
(15).—A solution of 12 (18.8 mg, 0.017 mmol) in THF
(2 mL) containing PtO2 (9.8 mg) was stirred under a
hydrogen atmosphere at rt for 16 h. The reaction
mixture was filtered and concentrated in vacuo. The
residue were dissolved in CHCl3 (6 mL), MeOH (12
mL) and 0.1 M aq HCl (4.8 mL). To the solution was
added another volume of CHCl3 (6 mL) and 0.1 M aq
HCl (6 mL) to separate the solution into two phases.
The lower CHCl3 phase was collected and concentrated
4,8-Anhydro-2,3,5-trideoxy-7-O-diphenylphosphono-5-
[(R) - 3 - (hydroxy)tetradecanoylamino] - 9 - O - methyl - 6-
O-[(R)-3-(tetradecanoyloxy)tetradecanoyl]-
D
-glycero-
D
-
to give 15 (16.0 mg, 98%), mp 211–213 °C. [h]D25
+
ido-nononic acid (13).—Compound 10 (67.4 mg) was
treated as described in the formation of 12 from 8 to
give 13 (49.8 mg, 83% yield). Rf 0.54 (1:9 MeOH–
CH2Cl2). IR wmax(neat) 3307, 2922, 2852, 1737, 1654,
49.5° (c 0.20, CHCl3). IR wmax(KBr) 3304, 2921, 2852,
1732, 1649, 1649, 1545, 1468, 1261, 1182, 1097, 1056
cm−1. 1H NMR (4:1 CD3OD–CDCl3): l 5.27–5.18 (m,
2 H), 4.30–4.22 (m, 2 H), 4.08 (m, 1 H), 3.94 (m, 1 H),
3.78 (br, 2 H), 3.64 (m, 1 H), 2.65 (d, 2 H, J 6.3 Hz),
2.49–2.24 (m, 6 H), 2.06 (m, 1 H), 1.87 (m, 1 H),
1.61–1.60 (m, 4 H), 1.45–1.29 (m, 58 H), 0.89 (m, 9 H).
13C NMR (100 MHz, CD3OD): l 176.8, 175.0, 174.2,
171.6, 74.0, 73.9, 73.4, 72.0, 71.2, 69.5, 61.8, 52.2, 44.4,
38.4, 35.2, 35.0, 32.8, 30.6, 30.2, 30.0, 29.9, 26.5, 26.5,
26.1, 25.9, 23.5, 21.6, 14.4. FABMS (positive-ion): m/z
978 [M+H]+, 1000 [M+Na]+. HRFABMS (positive-
ion): Calcd for C51H97NO14P: 978.6647. Found:
978.6631. Anal. Calcd for C51H96NO14P: C, 62.62; H,
9.89; N, 1.43; P, 3.17. Found: C, 62.61; H, 10.14; N,
1.24; P, 2.96.
1491, 1290, 1192, 1094, 1025, 960 cm−1 1H NMR
.
(CDCl3): l 7.36–7.17 (m, 10 H), 6.85 (d, 1 H, J 7.4 Hz),
5.30 (dd, 1 H, J 10.0 and 8.7 Hz), 5.08 (m, 1 H), 4.70
(dd, 1 H, J 17.5 and 8.8 Hz), 4.36–4.26 (m, 2 H), 3.94
(m, 1 H), 3.78 (m, 1 H), 3.46 (d, 2 H, J 3.5 Hz), 3.24 (s,
3 H), 2.54–2.20 (m, 8 H), 2.03–1.88 (m, 2 H), 1.58–
1.22 (m, 62 H) 0.88 (t, 9 H, J 6.6 Hz). 13C NMR (100
MHz, CD3OD): l 175.8, 173.9, 172.9, 171.2, 150.4,
129.9, 125.7, 120.2, 74.8, 72.3, 70.8, 70.7, 70.3, 70.0,
68.7, 59.2, 51.5, 42.8, 39.4, 37.2, 34.4, 34.3, 31.9, 29.7,
29.6, 29.5, 29.4, 29.2, 25.5, 25.1, 25.0, 22.7, 20.6, 14.1.
FABMS (positive-ion): m/z 1144 [M+H]+, 1166 [M+
Na]+. HRFABMS (positive-ion): Calcd for C64H106
-
4,8-Anhydro-2,3,5-trideoxy-5-[(R)-3-(hydroxy)tetra-
decanoylamino]-7-O-phosphono-9-O-methyl-6-O-[(R)-
NNaO14P: 1166.7248. Found: 1166.7263. Anal. Calcd
for C64H106NO14P: C, 67.16; H, 9.34; N, 1.22; P, 2.71.
Found: C, 67.32; H, 9.43; N, 1.14; P, 2.98.
4,8-Anhydro-2,3,5,9-tetradeoxy-7-O-diphenylphos-
phono-9-fluoro-5-[(R)-3-(hydroxy)tetradecanoylamino]-
3 - (tetradecanoyloxy)tetradecanoyl] -
D - glycero - D - ido-
nononic acid (16).—Compound 13 (38.3 mg) was
treated as described in the formation of 15 from 12 to
give 16 (33.6 mg, quant), mp 157–160 °C. [h]2D3 +18.4°
(c 0.20, CHCl3). IR wmax(neat) 3298, 2922, 2852, 1734,
6-O-[(R)-3-(tetradecanoyloxy)tetradecanoyl]-
D-glycero-
D
-ido-nononic acid (14).—Compound 11 (52.9 mg) was
1647, 1543, 1468, 1181, 1091 cm−1 1H NMR
.
treated as described in the formation of 12a from 8 to
give 12c (35.6 mg, 76% yield). Rf 0.55 (1:9 MeOH–
CH2Cl2). IR wmax(neat) 3307, 2925, 2854, 1736, 1655,
(CD3OD): l 5.25–5.17 (m, 2 H), 4.27–4.19 (m, 2 H),
4.03 (m, 1 H), 3.94 (m, 1 H), 3.78 (m, 2 H), 3.69 (dd, 1
H, J 10.7 and 2.4 Hz), 3.58 (dd, 1 H, J 10.8 and 5.9
Hz), 3.38 (s, 3 H), 2.65 (d, 2 H, J 6.2 Hz), 2.47–2.24
(m, 6 H), 2.04 (m, 1 H), 1.85 (m, 1 H), 1.60–1.59 (m, 4
H), 1.45–1.29 (m, 58 H) 0.90 (t, 9 H, J 6.8 Hz). 13C
NMR (100 MHz, CD3OD): l 176.9, 175.1, 174.4, 171.7,
1
1490, 1189, 961 cm−1. H NMR (CDCl3): l 7.36–7.16
(m, 10 H), 6.93 (d, 1 H, J 6.4 Hz), 5.32 (t, 1 H, J 9.3
Hz), 5.07 (m, 1 H), 4.64 (dd, 1 H, J 17.8 and 9.0 Hz),
4.46 (dd, 2 H, J 2.5, JFH 47.0 Hz), 4.34–4.29 (m, 2 H),

104
T. Wakabayashi et al. / Carbohydrate Research 337 (2002) 97–104
74.5, 73.6, 72.8, 72.4, 72.3, 71.4, 69.7, 59.5, 52.2, 44.8,
40.3, 38.6, 35.3, 35.2, 33.1, 30.8, 30.7, 30.5, 30.3, 26.8,
26.4, 26.2, 23.8, 22.3, 14.5. FABMS (positive-ion): m/z
992 [M+H]+, 1014 [M+Na]+, 1036 [M+2Na−H]+.
HRFABMS (positive-ion): Calcd for C52H98NNaO14P:
1014.6623. Found: 1014.6633. Anal. Calcd for
C52H98NO14P: C, 62.94; H, 9.96; N, 1.41; P, 3.12.
Found: C, 62.98; H, 9.84; N, 1.58; P, 3.11.
supernatant, the cells were incubated in 200 mL of fresh
RPMI-1640 medium containing 10% NBCS, 30 ng/mL
LPS and graded concentrations of compounds in a
humidified atmosphere of 5% CO2 for 4.5 h at 37 °C.
After incubation, the amounts of TNFa produced in the
culture supernatants were determined by the TNFa
ELISA kits. As a control, the amount of TNFa pro-
duced by U937 cells, which were stimulated with 30
ng/mL of LPS in the absence of compounds, was used.
TNFa production of U937 cells increased dose-depen-
dently until coming to 10 ng/mL of the LPS concentra-
tion, and reached to almost ceiling. Therefore, 30 ng/mL
of LPS for stimulation of U937 cells was used. All
experiments were carried out at least twice.
4,8-Anhydro-2,3,5,9-tetradeoxy-9-fluoro-5-[(R)-3-
(hydroxy)tetradecanoylamino] - 7 - O- p hosphono - 6 - O-
[(R) - 3 - (tetradecanoyloxy)tetradecanoyl] - D - glycero - D-
ido-nononic acid (17).—Compound 12c (33.2 mg) was
treated as described in the formation of 15 from 12 to
give 17 (27.1 mg, 94% yield), mp 148–167 °C. [h]D25
+
14.2° (c 0.21, CHCl3). IR wmax(neat) 3371, 2921, 2852,
1
1733, 1645, 1544, 1467, 1182, 1098 cm−1. H NMR
(CD3OD): l 5.28–5.17 (m, 2 H), 4.67–4.55 (m, 2 H, JFH
47.4 Hz), 4.29–4.19 (m, 2 H), 4.06 (m, 1 H), 3.94 (m, 1
H), 3.85 (m, 1 H, JFH 22.5 Hz), 2.66 (d, 2 H, J 6.3 Hz),
2.45–2.25 (m, 6 H), 2.07 (m, 1 H), 1.88 (m, 1 H), 1.61
References
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