Syntheses of hydantocidin and C-2-thioxohydantocidin

Article abstract of DOI:10.1016/S0008-6215(02)00217-3

Hydantocidin (12), a naturally occurring strong herbicide, was synthesized in 35.2% overall yield, with accompanying 5-epi-hydantocidin (12′) in 9.6% overall yield via isothiocyanate (13) and spiro-hydantoin (10) from 2,3-O-isopropylidene-D-ribono-1,4-lactone (1). C-2-Thioxo-hydantocidin (24) was also synthesized in 16.5% overall yield with accompanying 5-epi-C-2-thioxohydantocidin (24′, 9.2% yield) via isothiocyanate (22).

Full text of DOI:10.1016/S0008-6215(02)00217-3

Carbohydrate Research 337 (2002) 2077–2088
www.elsevier.com/locate/carres
Syntheses of hydantocidin and C-2-thioxohydantocidin
Masao Shiozaki*
Exploratory Chemistry Research Laboratories, Sankyo Co., Ltd., Hiromachi 1-2-58, Shinagawa-ku, Tokyo 140-8710, Japan
Received 27 February 2002; accepted 22 May 2002
Dedicated to Professor Derek Horton on the occasion of his 70th birthday
Abstract
Hydantocidin (12), a naturally occurring strong herbicide, was synthesized in 35.2% overall yield, with accompanying
5-epi-hydantocidin (12%) in 9.6% overall yield via isothiocyanate (13) and spiro-hydantoin (10) from 2,3-O-isopropylidene- -ri-
D
bono-1,4-lactone (1). C-2-Thioxo-hydantocidin (24) was also synthesized in 16.5% overall yield with accompanying 5-epi-C-2-
thioxohydantocidin (24%, 9.2% yield) via isothiocyanate (22). © 2002 Elsevier Science Ltd. All rights reserved.
Keywords: Spironucleoside; Furanoid spirohydantoin; Glycosyl isothiocyanate; Herbicide
tant weeds. However, the high cost of hydantocidin
production, whether by fermentation or by total syn-
thesis,3 has made its use as a herbicide impractical.
Therefore, economical production of hydantocidin is
being sought. At this time, the author has accomplished
a fairly good overall yield for hydantocidin, that is,
hydantocidin 12 was synthesized in 35.2% overall yield
accompanying epi-hydantocidin 12% in 9.6% overall
1. Introduction
Together with the explosive increase in the world
population, deficiency of provisions, environmental de-
struction and pollution, and global warming have be-
come serious problems. Means for grain production to
maintain this large population have come under in-
creased development. One aspect is the use of herbi-
cides. Glyphosate has become one of the most popular
herbicides in the world. However, glyphosate-resistant
weeds have been reported recently. In addition, proteins
yield from 2,3-O-isopropylidene-D-ribono-1,4-lactone
(1). And also C-2-thioxohydantocidin 24 was synthe-
sized from 1 in 16.5% overall yield via isothiocyanate
22. Therefore, I would like to report the synthetic
procedure in detail here.4
produced by grain plants recombinated by
a
glyphosate-resistant gene are a cause for concern, be-
cause of the awareness of the existence of pathogenic
proteins.
Hydantocidin produced from Streptomyces hygro-
scopicus,1 the first naturally occurring spiro-ribofura-
nose having strong herbicidal activity toward annual,
biennial and perennial weeds by action as an adenylo-
succinate synthetase inhibitor2 without showing toxicity
to microorganisms and animals and without remaining
for a long period in the soil, may be used in the near
future as a potential herbicide against glyphosate-resis-
2. Results and discussion
The starting material, 2,3-O-isopropylidene-D-ri-
bono-1,4-lactone (1),5 was converted to 5-O-benzyl
ether 2 or 5-O-(4-methoxybenzyl) ether 15 in 95 or 77%
yield, respectively, by treatment with benzyl bromide or
4-methoxybenzyl chloride using NaH as a base. Treat-
ment of
2
with CBrCl3 using tris(dimethy-
lamino)phosphine [(Me2N)3P] as a base according to
Chapleur’s procedure6 gave dichloroolefin 37 in 86%
yield. Compound 15 also gave 16 in 95% yield. In this
reaction, Lakhrissi and Chapleur6 used CCl4 instead of
CBrCl3. However, in the case of using CCl4, the reac-
* Present address: Chemistry Department, Chemtech Labo,
Inc., Hiromachi 1-2-58, Shinagawa-ku, Tokyo 140-8710,
Japan; tel.: +81-3-34923131; fax: +81-3-54368570
E-mail address: shioza@shina.sankyo.co.jp (M. Shiozaki).
0008-6215/02/$ - see front matter © 2002 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(02)00217-3

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M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
4-methoxybenzyl isocyanate. Therefore, another syn-
thetic route via isothiocyanates 13 and 22 was investi-
gated (Scheme 3).
tion, on occasion, did not proceed. The dichloroolefin 3
was further converted to methyl a-chlorourosonates 4
and 4% in 68% yield as a 4:1 diastereomeric mixture via
C-1ꢀC-2 epoxide by treatment with 3-chloroperoxyben-
zoic acid.6 The mixture of 4 and 4% was separated
chromatographically. The configuration of 4 was deter-
mined by measurement of the nuclear Overhauser effect
(NOE) between H-3 (the proton attached to the 3-posi-
tion carbon) and 13C-1 (methyl ester carbonyl carbon)
of compounds 4 and 4%. The NOE between H-3 and
13C-1 of 4 was detected. However, the NOE between
H-3–13C-1 of 4% was not detected. This result indicates
that the configurations of the carbonyl carbon of the
C-1 methyl ester and the C-3 proton in compound 4 are
on the same side of the molecule, while those of 4% are
trans to one another. Compound 16 was also converted
to a 4:1 mixture of 17 and 17% (epimer of 17) in 67%
yield in the same treatment. The mixture of 17 and 17%
was separated chromatographically.
Firstly, a route through carbodiimide 7 and ureido 9
was attempted. Treatment of 4 with sodium azide in
DMF at room temperature gave azide 5 in 95% yield
with inversion of configuration. Conversion of 5 to
phosphine imide 6 by treatment of triphenylphosphine,
and successive treatment of 6 with 4-mehoxybenzyl
isocyanate yielded carbodiimide 7 (90%) as a single
product. Treatment of 7 with aq 1 M HCl in THF
afforded urea 8 quantitatively. Deprotection of the
4-methoxybenzyl group from 8 was carried out by
treatment with ammonium cerium(IV) nitrate (CAN) to
give 9 in 97% yield. Hydantoin formation from 9 with
NH3 in MeOH proceeded smoothly to give spiro-hy-
dantoin 10 in 99% yield. Hydrogenolysis of 10 using
Pd-on-charcoal afforded 11 quantitatively. Treatment
of 11 with 1:3 CF3COOH–water at 0 °C according to
the reported method3f yielded hyantocidin 12 quantita-
tively. This synthetic hydantocidin was identical with
natural hydantocidin in all respects, including herbici-
dal activity toward many kinds of weeds (Schemes 1
and 2).
In this second route, treatment of 4 with KSCN at
80 °C in DMF gave a 4:1 mixture of isothiocyanate 13
and 13% in 84% yield without yielding corresponding
thiocyanates.4,8,9 Similarly, the same treatment of 4%
gave a 5:2 mixture of 13 and 13% in 69% yield. Separa-
tion of isothiocyanates 13 and 13% was difficult by
chromatography. However, a sample of the mixture
was partially separated on a TLC plate by development
with 3:1 cyclohexane–EtOAc (13, Rf 0.500; 13%, Rf
0.467) to measure the physical data. The 13C NMR
spectra of 13 and 13% indicated the presence of the
isothiocyanates (l 143.2 and 144.4 ppm, respectively).9
This mixture was treated with NH3 in MeOH to give
C-2-thioxohydantoins 14 and 14%, quantitatively. The
ratio of 14 and 14% was in parallel with that of the
starting 13 and 13%. And the same reaction of com-
pounds 17 and 17% also gave a 5:1 mixture of corre-
sponding isothiocyanates 18 and 18% in 74% yield from
17, and a 3:1 mixture of 18 and 18% in 72% yield from
17%. These compounds 18 and 18% were also inseparable
by chromatography. And the mixture of 18 and 18% was
also treated by NH3 to give hydantoins 19 and 19%
quantitatively in the same ratio as that of the starting
18 and 18%. These 4-methoxybenzyl-protected spiro-hy-
dantoins 19 and 19% were also used for unnatural C-2-
thioxohydantocidin 24 as mentioned later. Treatment
of thioxo-hydantoins 14 and its spiro epimer 14% with
H2O2–aq NaHCO3 quantitatively gave hydantoin 10
and its epimer 10%, respectively.10 And thioxo-com-
pounds 19 and 19% were also converted to oxo-com-
pounds 20 and 20% in 96 and 95% yields, respectively.
This procedure was also applicable for the oxidation of
thiourea, thiohydantoin and cyclic thiocarbamate to
yield corresponding urea, hydantoin and carbamate as
reported in a communication.4
Spiro-hydantoin 10 and its spiro epimer 10% were
converted quantitatively to hydantocidin 12 and its
spiro epimer 12% in two steps as mentioned above. And
compounds 20 and 20% were also converted quantita-
tively to hydantocidin 12 and 12% via the hy-
drogenolyzed compounds 11 and 11%, respectively.
Thus, hydantocidin 12 was synthesized in eight steps
from 1 via isothiocyanate 13, and the overall yield was
35.2% accompanied by 9.6% yield of spiro-epi-hydanto-
cidin 12%. Also, hydantocidin was synthesized in eight
steps from 1 via isothiocyanates 18 and 18% in 30.0%
overall yield accompanied by spiro-epi-hydantocidin 12%
(6.3%). The yield of this route was lower than that of
the route via 13. Besides, the 4-methoxybenzyl group is
much more expensive than the benzyl group for synthe-
sis. In either case, the difference is little, that is, one is
benzyl, and the other is 4-methoxybenzyl.
The spiro-epi-hydantocidin (12%) was also synthesized
by the same procedure as for the synthesis of 12 from
methyl a-chlorourosonate 4% through the corresponding
epimeric intermediates 5%–11%. In the preparation of
ureido 8% (67% yield) from 7%, a fair amount of 8%% (14%)
was obtained by acidic cleavage of the acetonide pro-
tecting group. Compound 10% was also obtained in 91%
yield from 9 by both epimerization and hydantoin
formation using 1,8-diazabicyclo[5.4.0]undec-7-ene
(DBU). Necessarily, compound 10 was epimerized to
10% in 87% yield (with 12% recovery of 10) using the
same conditions.
However, this route was not economical because of
the high cost of the reagents for agricultural chemicals
used, such as sodium azide, triphenylphosphine and

M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
2079
Scheme 1. Reagents and conditions: Bn=benzyl; PMB=4-methoxybenzyl; (a) NaH, BnBr, or PMB–Cl, DMF, 24 °C, 16 h, 95%
(2) and 77% (15); (b) CBrCl3, (Me2N)3P, CH2Cl2, −78–24 °C, 16 h, 86% (3) and 95% (16); (c) m-CPBA, MeOH, CH2Cl2, 24 °C,
16 h, 54% (4) and 14% (4%), and 54% (17) and 13% (17%); (d) NaN3, DMF, 24 °C, 16 h, 95%; (e) PPh3, THF, 24 °C, 2 h; (f)
PMBꢀNꢁCꢁO, THF, 24 °C, 16 h, two steps 90%; (g) 1:20 aq 1 M HCl–THF, rt, 15 min, quant; (h) Ce(NH4)2(NO3)6, 2:1
MeCN–water, rt, 20 min, 97%; (i) 0.2 M NH3 in MeOH, 27 °C, 4 h, 99%; (j) H2, Pd/C, EtOAc, 24 °C, 30 min, quant; (k) (Ref.
3f) 1:3 CF3COOH–water, 0 °C, 2 h, quant.
Thirdly, C-2-thioxohydantocidin 24, which has been
reported along with its spiro epimer 24% as being com-
parable with hydantocidin 12 in herbicidal activity to
many different types of weeds,3k,11 is not found in
nature and, therefore, has to be synthesized.
Methyl a-chlorourosonate 4 was hydrogenolyzed to
deprotect the benzyl group, and the resulting alcohol
was acetylated with acetic anhydride and pyridine to
give 21 in 90% yield. On the other hand, compound 4%
just decomposed to unknown materials in the same
procedure without yielding 21%. However, considering
that treatment of 4% itself at 80 °C for 1 h in DMF gave
ca. 1:1 mixture of 4 and 4% by thermal equilibration, this
problem (unavailability of 4%) should be resolved in
further studies. Methyl a-chlorourosonate 21 was also
converted to isothiocyanate 22 (68%) and its epimer 22%
(13%) by treatment with potassium thiocyanate
(KSCN) at 80 °C in DMF. The 13C NMR spectra of 22
and 22% showed resonances for isothiocyanate groups (l
144.5 and 145.0 ppm, respectively).9 Reaction of 22 and
22% with NH3 in MeOH gave the deacetylated spiro-hy-
dantoins 23 and 23% quantitatively, respectively. On the
other hand, treatment of 19 and 19% in CH2Cl2 with
PhSH and SnCl4 at −78 °C under N2 gave 23 and 23%

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M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
Scheme 2. Reagents and conditions: Bn=benzyl; PMB=4-methoxybenzyl; (d) NaN3, DMF, 24 °C, 16 h, 89%; (e) PPh3, THF,
24 °C, 2 h; (f) PMBꢀNꢁCꢁO, THF, 24 °C, 16 h, two steps 69%; (g) 1:20 aq 1 M HCl–THF, rt, 15 min, 67% (8%) and 14% (8%%);
(h) Ce(NH4)2(NO3)6, 2:1 MeCN–water, rt, 20 min, 72%; (i) 0.2 M NH3 in MeOH, 27 °C, 4 h, quant; (j) H2, Pd/C, EtOAc, 24 °C,
30 min, quant; (k) (Ref. 3f) 1:3 CF3COOH–water, 0 °C, 2 h, quant; (l) DBU, THF–DMF, rt, 7 days, 91% from 9, and 87% from
10 (12% recovery of 10).
in 81 and 72% yields, respectively, accompanying recov-
ery of 19 (6%) and 19% (13%), respectively.12 Finally, the
protecting isopropylidene group of 23 was cleaved by
treatment with 3:1 CF3COOH–water at 0 °C for 2 h
and −5 °C for 16 h to give a chromatographically
separable mixture of 24 (61%) and 24% (16%).3f How-
ever, the same treatment of 23% only gave 24% in 94%
yield. As a result, C-2-thioxohyantocidin 24 was synthe-
sized from 1 in eight steps via the acetylated isothio-
cyanates 22 and 22% in 16.5% overall yield, along with
the accompanying spiro epimer 24% (9.2% yield). These
compounds were also synthesized from 1 in seven steps
via isothiocyanate intermediates 19 and 19% in 15.4%
overall yield, along with the accompanying spiro epimer
24% (4.2% yield). The yield of this route was a little
lower than that of the route via 22 (Scheme 4).
with a JEOL-GSX 400 spectrometer (400 MHz) using
TMS as the internal standard, and 13C NMR spectra
were recorded at 125 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. Separa-
tion of the compounds by column chromatography was
done 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 quantity of silica gel used was
50–100 times the weight charged on the column. Thin-
layer chromatography was performed on E. Merck no.
5715 Silica Gel 60-F254 plates. Tetrahydrofuran (THF)
was distilled in the presence of radical anions generated
by sodium-benzophenone ketyl. Dichloromethane was
dried by being passed through an ICN Alumina B-Su-
per I, and DMF and pyridine were dried by storage
Thus, hydantocidin 12 and spiro-epi-hydantocidin
,
12% were synthesized from 2,3-O-isopropylidene-
D
-ri-
over 4 A molecular sieves.
5-O-Benzyl-2,3-O-isopropylidene-
bono-1,4-lactone 1 in overall yields of 35.2 and 9.6%
via isothiocyanate intermediates 13 and 13%, and 30.0
and 6.3% via intermediates 19 and 20, and 19% and 20%,
respectively. C-2-Thioxohydantocidin 24 and spiro-epi-
C-2-thioxohydantocidin 24% were synthesized from 1 in
overall yields of 16.5 and 9.2% via isothiocyanate inter-
mediates 22 and 22%, and 15.4 and 4.2% via intermedi-
ates 19 and 19%.
D
-ribono-1,4-lac-
-ri-
tone (2).—To a solution of 2,3-O-isopropylidene-
D
bono-1,4-lactone (1, 7.45 g, 39.58 mmol) and benzyl
bromide (8.12 g, 47.50 mmol) in DMF (30 mL) was
added NaH (60–72% oil dispersion, 1.90 g) under ice
cooling. The mixture was stirred at 0 °C for 30 min,
and then rt for 16 h. The mixture was then diluted
with EtOAc, washed with aq 0.1 M HCl, satd aq
NaHCO3, and brine, then dried over MgSO4, filtered,
and concentrated in vacuo to give an oily mixture. The
mixture was chromatographed on a silica gel column.
Elution with 3:1 cyclohexane–EtOAc gave 2 (10.45 g,
3. Experimental
1
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
95%). H NMR (CDCl3): l 1.37 (3 H, s), 1.47 (3 H, s),
3.68 (1 H, dd, J 1.5, 11.0 Hz), 4.72 (1 H, dd, J 2.2, 11.0
Hz), 4.47, 4.56 (2 H, AB-q, J 11.7 Hz), 4.65 (1 H, dd,
J 1.5, 2.2 Hz), 4.71 (1 H, d, J 5.9 Hz), 4.79 (1 H, d, J

M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
2081
ture, which was chromatographed on a silica gel
column. Elution with 3:1 cyclohexane–EtOAc gave 3
(6.37 g, 86%) as an oil and starting 2 (0.45 g, 7.5%
recovery). IR wmax(film) 1765, 1731 cm−1 1H NMR
.
(CDCl3): l 1.39 (3 H, s), 1.49 (3 H, s), 3.65 (2 H, s),
4.51 (2 H, s), 4.65 (1 H, s), 4.80, 5.26 (2 H, AB-q, J 6.0
Hz), 7.27–7.40 (5 H, m). FABMS (positive-ion) m/z:
344 [M+ ]. HRFABMS (positive-ion), Calcd for
’
C16H18 35Cl2O4: 344.0582; Found: 344.0599. Anal.
Calcd for C16H18Cl2O4: C, 55.67; H, 5.26; Cl, 20.54.
Found: C, 55.94; H, 5.16; Cl, 20.11.
Methyl 2,5-anhydro-6-O-benzyl-2-chloro-3,4-O-iso-
propylidene-h-
methyl 2,5-anhydro-6-O-benzyl-2-chloro-2-deoxy-3,4-
O-isopropylidene-i- -ribo-2-hexulofuranosonate (4%).—
D-ribo-2-hexulofuranosonate
(4)
and
D
To a solution of 3 (345 mg, 1.00 mmol) in CH2Cl2 (20
mL) containing MeOH (300 mg) were added hy-
droquinone (50 mg) and 4-chloroperoxybenzoic acid
(1.20 g, purity ca 77%, Aldrich Chemical Co.) under N2
at rt. The mixture was stirred for 16 h at rt, and then
the reaction mixture was diluted with CH2Cl2 and
washed with 10% aq NaHSO3 (two times), satd aq
NaHCO3, and brine. The organic layer was dried over
MgSO4, filtered, and concentrated in vacuo to give a
residue, which was chromatographed on a silica gel
column. Elution with 3:1 cyclohexane–EtOAc gave 2-
epimer 4% (0.51 g, 14%, Rf 0.508) as an amorphous solid
and 4 (1.94 g, 54%, Rf 0.398) as a gum. Physical data of
4%: mp 48–48.5 °C (from hexane); [h]2D5 −99.5° (c 0.5,
1
CHCl3). IR wmax(KBr) 1762 cm−1. H NMR (CDCl3):
l 1.33 (3 H, s), 1.44 (3 H, s), 3.75 (2 H, d, J 7.3 Hz,
C-6ꢀH2), 3.89 (3 H, s), 4.58, 4.60 (2 H, AB-q, J 12.5
Hz), 4.66 (1 H, dt, J 2.2, 7.3 Hz, C-5ꢀH), 4.93 (1 H, dd,
J 1.5, 5.9 Hz, C-4ꢀH), 5.14 (1 H, d, J 5.9 Hz, C-3ꢀH),
7.27–7.35 (5 H, m). 13C NMR (100 MHz, CDCl3):
25.44, 26.25, 53.48 (OCH3), 68.81, 73.30, 81.71, 88.51,
89.80, 103.09, 114.14, 127.63 (3 C), 128.31 (2 C),
137.35, 164.89. NOE between H-3 and 13C-1 was not
detected. FABMS (positive-ion) 355 [M−H]+, 357
[M+H]+, 379 [M+Na]+ (on addition of aq NaI).
HRFABMS (positive-ion), Calcd for C17H21ClNaO6:
379.0924; Found: 379.0940. Physical data of 4: [h]D23
+41.4° (c 3.2, CHCl3). IR wmax(film) 3089–2867, 1750
Scheme 3. Reagents and conditions: Bn=benzyl; PMB=4-
methoxybenzyl; (a) KSCN, DMF, 80 °C, 5 h, 84% (a 4:1
mixture of 13 and 13% from 4), 69% (a 5:2 mixture of 13 and
13% from 4%), 73% (a 5:1 mixture of 18 and 18% from 17), and
72% (a 3:1 mixture of 18 and 18% from 17%); (b) NH3, MeOH,
rt, 30 min, quant; (c) H2O2, NaHCO3, MeCN–water, rt, 15
min, quant (10 and 10%), 96% (20), and 95% (20%); (d) H2,
Pd/C, EtOAc, rt, quant.
5.9 Hz), 7.24–7.38 (5 H, m). FABMS (positive-ion)
m/z: 279 [M+H]+.
2,5-Anhydro-6-O-benzyl-1,1-dichloro-1-deoxy-3,4-O-
1
cm−1. H NMR (CDCl3): l 1.40 (3 H, s), 1.70 (3 H, s),
3.69 (1 H, dd, J 3.2, 11.3 Hz, C-6ꢀH), 3.73 (1 H, dd, J
3.2, 11.3 Hz, C-6ꢀH), 3.83 (3 H, s), 4.53, 4.58 (2 H,
AB-q, J 12.2 Hz), 4.59 (1 H, q, J 3.2 Hz, C-5ꢀH), 4.81
(1 H, dd, J 3.2, 7.1 Hz, C-4ꢀH), 5.02 (1 H, d, J 7.1 Hz,
C-3ꢀH), 7.28–7.39 (5 H, m). 13C NMR (100 MHz ,
CDCl3): 25.29 (2 C), 53.48 (OCH3), 68.58, 73.41, 79.80,
83.68 (2 C), 102.44, 116.78, 127.55 (2 C), 127.75, 128.31
(2 C), 137.21, 166.22. NOEs between H-3 and 13C-1,
H-3 and 13C-2, H-3 and 13C-4, H-3 and 13C-5, and H-3
and 13C-benzylic were detected. Anal. Calcd for
C17H21ClO6: C, 57.23; H, 5.93; Cl, 9.94. Found: C,
57.01; H, 5.77; Cl, 10.14.
isopropylidene- -ribo-hex-1-enitol (3).—To a solution
D
of 2 (6.00 g, 21.56 mmol) and BrCCl3 (21.37 g, 107.80
mmol) in CH2Cl2 (60 mL) was added dropwise a solu-
tion of P(NMe2)3 (35.20 g, 46.08 mmol) in CH2Cl2 (40
mL) at −78 °C over 1.5 h under nitrogen. The temper-
ature was gradually elevated to −40 °C during 4 h
stirring, and then to rt with stirring for 16 h. The
reaction mixture was diluted with CH2Cl2, washed with
aq 0.5 M HCl, water, aq NaHCO3, and brine, and the
organic layer was then dried over MgSO4. The solution
was filtered and concentrated in vacuo to give a mix-

2082
M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
Scheme 4. Reagents and conditions: (a) (i) H2, Pd/C, EtOAc, rt, 20 min; (ii) Ac2O, pyridine, EtOAc, rt, 15 h, 2 steps 90%; (b)
KSCN, DMF, 80 °C, 15 h, 68% (22) and 13% (22%); (c) NH3 in MeOH, rt, 6 h, quant; (d) SnCl4, PhSH, CH2Cl2, −78 °C, 30 min,
81% (23), or 76% (23%) and 13% (recovery of 19%); (e) 1:3 CF3COOH–water, 0 °C, 3 h, 68% (24) and 13% (24%) from 23, or 20 °C,
2 h, 94% (24%) from 23%.
Methyl 2,5-anhydro-2-azido-6-O-benzyl-3,4-O-iso-
propylidene-i- -ribo-2-hexulofuranosonate (5).—To a
d, J 6.2 Hz), 7.26–7.34 (5 H, m). FABMS (positive-ion)
m/z: 362 [M−H]+, 386 [M+Na]+.
D
solution of 4 (835 mg, 2.34 mmol) in DMF (10 mL)
was added NaN3 (457 mg, 7.02 mmol). After 5 h
stirring at rt, the reaction mixture was diluted with
EtOAc, washed with water, and brine. The organic
extract was dried over MgSO4, filtered, and then con-
centrated in vacuo (finally with a pump) to give 5 (808
Methyl
2,5-anhydro-6-O-benzyl-3,4-O-isopropyli-
-ribo-
dene-2-[3-N-(4-methoxybenzyl)carbodiimido]-i-
D
2-hexulofuranosonate (7).—To a solution of 5 (760 mg,
2.09 mmol) in THF (35 mL) was added PPh3 (1.20 g,
4.57 mmol) to yield phosphine imide 6. After 2 h
stirring at rt, 4-methoxybenzyl isocyanate (1.00 g, 6.13
mmol) was added to this solution. The mixture was
stirred for 3 days at rt. The reaction mixture was
concentrated in vacuo to give a mixture that was chro-
matographed on a silica gel column. Elution with 2:1
cyclohexane–EtOAc gave carbodiimide 7 (904 mg,
90%) as an oil. IR wmax(film) 3063–2840, 2130, 1767,
1
mg, 95%). IR wmax(film) 2117, 1764 cm−1. H NMR
(CDCl3): l 1.30 (3 H, s), 1.45 (3 H, s), 3.63 (1 H, dd, J
6.2, 10.1 Hz, C-6ꢀH), 3.65 (1 H, dd, J 5.2, 10.1 Hz,
C-6ꢀH), 3.87 (3 H, s), 4.56, 4.61 (2 H, AB-q, J 12.0
Hz), 4.63–4.67 (2 H, m), 4.87 (1 H, dd, J 1.5, 5.8 Hz),
7.34–7.36 (5 H, m). FABMS (positive-ion) m/z: 386
[M+Na]+ (on addition of NaI). Anal. Calcd for
C17H21N3O6: C, 56.19; H, 5.83; N, 11.57. Found: C,
56.02; H, 5.76; N, 11.50.
1745, 1612 cm−1. H NMR (CDCl3): l 1.29 (3 H, s),
1
1.44 (3 H, s), 3.60–3.74 (2 H, m), 3.78 (6 H, s), 4.10 (1
H, m), 4.19 (1 H, m), 4.37, 4.52 (2 H, AB-q, J 11.5 Hz),
4.58 (1 H, s), 4.69 (1 H, m), 4.85 (1 H, m), 6.81–6.84 (2
H, m), 7.10–7.12 (2 H, m), 7.26–7.27 (5 H, m).
FABMS (positive-ion) m/z: 483 [M+H]+. HRFABMS
(positive-ion), Calcd for C26H31N2O7: 483.2131; Found:
483.2153. Anal. Calcd for C26H30N2O7: C, 64.72; H,
6.27; N, 5.81. Found: C, 64.36; H, 6.37; N, 5.75.
Methyl 2,5-anhydro-2-azido-6-O-benzyl-3,4-O-iso-
propylidene-h- -ribo-2-hexulofuranosonate (5%).—Com-
D
pound 5% was also obtained in 89% yield as an oil from
4% according to the same procedure described above. IR
wmax(film) 3065–2866, 2127, 1751 cm−1 1H NMR
.
(CDCl3): l 1.38 (3 H, s), 1.65 (3 H, s), 3.61–3.63 (2 H,
m), 3.70 (3 H, s), 4.45, 4.54 (2 H, AB-q, J 12.0 Hz),
4.52 (1 H, m), 4.79 (1 H, dd, J 1.6, 6.2 Hz), 5.13 (1 H,
Methyl
2,5-anhydro-6-O-benzyl-3,4-O-isopropyli-
-ribo-
dene-2-[3-N-(4-methoxybenzyl)carbodiimido]-h-
D

M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
2083
2-hexulofuranosonate (7%).—Compound 7% was also ob-
tained in 69% yield from 5% via phosphazene 6% accord-
ing to the same procedure described above. IR
Calcd for C26H33N2O8: 501.2237; Found: 501.2233.
Physical data of 8%%: IR wmax(CHCl3) 3350, 3365–2840,
1
1746, 1654, 1612, 1587, 1514 cm−1. H NMR (CDCl3):
1
wmax(film) 3063–2838, 2134, 1744, 1613, 1587 cm−1. H
l 3.69–3.72 (3 H, m), 3.76 (6 H, s), 4.14 (1 H, m), 4.34
(2 H, s), 4.53, 4.56 (2 H, AB-q, J 11.2 Hz), 5.05 (1 H,
d, J 5.9 Hz), 6.82–6.85 (2 H, m), 7.19–7.22 (2 H, m),
7.25–7.35 (5 H, m). FABMS (positive-ion) m/z: 443
[M+H]+, 481 [M+K]+ (on addition of KI). HR-
FABMS (positive-ion), Calcd for C23H26KN2O7:
481.1377; Found: 481.1374. Anal. Calcd for
C23H26N2O7: C, 62.43; H, 5.92; N, 6.33. Found: C,
62.21; H, 6.13; N, 6.20.
NMR (CDCl3): l 1.37 (3 H, s), 1.61 (3 H, s), 3.59–3.61
(2 H, m), 3.64 (3 H, s), 3.79 (3 H, s), 4.43–4.57 (5 H,
m), 4.75 (1 H, dd, J 2.2, 6.6 Hz), 5.02 (1 H, d, J 6.6
Hz), 6.84–6.87 (2 H, m), 7.24–7.36 (7 H, m). FABMS
(positive-ion) m/z: 483 [M+H]+. HRFABMS (posi-
tive-ion), Calcd for C26H31N2O7: 483.2131; Found:
483.2126.
Methyl
2,5-anhydro-6-O-benzyl-3,4-O-isopropyli-
-ribo-2-hex-
dene-2-[3-N-(4-methoxybenzyl)ureido]-i-
D
Methyl
dene-2-ureido-i-
2,5-anhydro-6-O-benzyl-3,4-O-isopropyli-
-ribo-2-hexulofuranosonate (9).—To
ulofuranosonate (8).—To a solution of 7 (452 mg, 0.94
mmol) in THF (20 mL) was added aq 1 M HCl (1.00
mL) at rt. After stirring for 40 min, the reaction
mixture was diluted with EtOAc. The solution was
washed with satd aq NaHCO3 and brine, and the
organic layer was dried over MgSO4, and filtered. The
filtrate was concentrated in vacuo to give ureido 8 (469
mg, quant) as a solid; mp 149–151 °C (from EtOAc).
[h]2D5 −84° (c 0.05, CHCl3). IR wmax(KBr) 3350, 3088–
D
a solution of 8 (230 mg, 0.46 mmol) in MeCN (20 mL)
was added a solution of ammonium cerium(IV) nitrate
(4.11 g, 7.50 mmol) in water (10 mL) at rt with stirring.
After 20 min, the reaction mixture was diluted with
EtOAc, washed with sat aq NaHCO3, and brine. The
organic layer was dried over MgSO4, filtered, and con-
centrated in vacuo to give a mixture, which was chro-
matographed on a silica gel column. Elution with 3%
MeOH in EtOAc gave 9 (170 mg, 97%) as a solid; mp
201–202 °C (from MeOH). IR wmax(KBr) 3365, 3207,
1
2838, 1754, 1656, 1614, 1564, 1513 cm−1. H NMR
(CDCl3): l 1.29 (3 H, s), 1.44 (3 H, s), 3.65 (1 H, dd, J
4.1, 10.3 Hz), 3.69 (1 H, dd, J 3.8, 10.3 Hz), 3.77 (3 H,
s), 3.78 (3 H, s), 4.10 (1 H, dd, J 4.8, 14.6 Hz, changed
to a doublet, J 14.6 Hz, on addition of D2O), 4.19 (1 H,
dd, J 5.8, 14.6 Hz, changed to a doublet, J 14.6 Hz, on
addition of D2O), 4.37, 4.51 (2 H, AB-q, J 11.4 Hz),
4.58 (1 H, m), 4.69 (1 H, d, J 6.2 Hz), 4.85 (1 H, m,
changed to dd, J 2.2, 6.2 Hz, on addition of D2O), 5.20
(1 H, bs, NH, exchanged on addition of D2O), 5.65 (1
H, s, NH, not exchanged on addition of D2O), 6.82 (2
H, d, J 8.3 Hz), 7.11 (2 H, m), 7.19–7.21 (2 H, m),
7.26–7.30 (3 H, m). FABMS (positive-ion) m/z: 501
[M+H]+, 523 [M+Na+]. HRFABMS (positive-ion),
Calcd for C26H32N2NaO8: 523.2057; Found: 523.2065.
Anal. Calcd for C26H32N2O8: C, 62.39; H, 6.44; N, 5.60.
Found: C, 62.28; H, 6.10; N, 5.55.
3088–2864, 2557–2385, 1751, 1658 cm−1 1H NMR
.
(CDCl3): l 1.31 (3 H, s), 1.45 (3 H, s), 3.68 (1 H, dd, J
4.1, 10.4 Hz), 3.74 (1 H, dd, J 4.1, 10.4 Hz), 3.80 (3 H,
s), 4.49, 4.57 (2 H, AB-q, J 11.4 Hz), 4.59 (1 H, m),
4.81 (1 H, d, J 6.4 Hz), 4.90 (1 H, dd, J 2.5, 6.4 Hz),
4.94 (2 H, bs, NH2, exchanged on addition of D2O),
6.23 (1 H, s, NH, not exchanged on addition of D2O),
7.28–7.40 (5 H, m). FABMS (positive-ion) m/z: 381
[M+H]+, 403 [M+Na+]. HRFABMS (positive-ion),
Calcd for C18H25N2O7: 381.1662; Found: 381.1668.
Anal. Calcd for C18H24N2O7: C, 56.83; H, 6.36; N, 7.36.
Found: C, 56.29; H, 6.78; N, 6.92.
Methyl
2,5-anhydro-6-O-benzyl-3,4-O-isopropyli-
dene-2-ureido-h-D-ribo-2-hexulofuranosonate (9%).—
Compound 9%, accompanied by a small amount of
chromatographycally inseparable 4-epimer (9), was also
obtained from 8% in 72% yield according to the same
procedure described above. IR wmax(CHCl3) 4214, 3514,
Methyl
2,5-anhydro-6-O-benzyl-3,4-O-isopropyli-
-ribo-2-hex-
dene-2-[3-N-(4-methoxybenzyl)ureido]-h-
D
ulofuranosonate (8%).—Compound 8% was also obtained
in 67% yield as a powder by treatment of 7% for 2 h,
instead of 40 min according to the procedure as de-
scribed above, and a byproduct (8%%) was obtained in
14% yield. Physical data of 8%: IR wmax(CHCl3) 4214,
3403, 3020–2860, 1755, 1690, 1594 cm−1 1H NMR
.
(CDCl3): l 1.39 (3 H, s), 1.63 (3 H, s), 3.64–3.70 (5 H,
m, containing 3 H, s, at d 3.68 ppm), 4.41 (1 H, m),
4.48–4.58 (2 H, m), 4.77–4.88 (2 H, m), 5.10 (2 H, bs,
NH2), 5.83 (1 H, s, NH), 7.29–7.37 (5 H, m). FABMS
(positive-ion) m/z: 381 [M+H]+, 403 [M+Na+]. HR-
FABMS (positive-ion), Calcd for C18H25N2O7:
381.1662; Found: 381.1642.
[5S-(5h,7h,8i,9i)]-7-(Benzyloxymethyl)-8,9-isopro-
pylidenedioxy - 6 - oxa - 1,3 - diazaspiro[4.4]nonane - 2,4-
dione (10).—(a) To a solution of 9 (48.0 mg, 0.13
mmol) in MeOH (5 mL) was added a solution of 2 M
NH3 in MeOH (0.5 mL) at 37 °C with stirring. After 4
h, the reaction mixture was concentrated in vacuo to
3411, 3088–2839, 1753, 1678, 1613 cm−1 1H NMR
.
(CDCl3): l 1.39 (3 H, s), 1.62 (3 H, s), 3.62 (3 H, s),
3.64 (1 H, dd, J 4.4, 11.0 Hz), 3.70 (1 H, dd, J 4.4, 11.0
Hz), 3.79 (3 H, s), 4.26 (1 H, dd, J 5.9, 14.6 Hz),
4.37–4.42 (2 H, m), 4.49, 4.57 (2 H, AB-q, J 12.1 Hz),
4.78 (1 H, dd, J 2.9, 6.6 Hz), 4.83 (1 H, d, J 7.3 Hz),
5.66 (1 H, bs, NH, exchanged on addition of D2O), 5.73
(1 H, s, NH), 6.83–6.85 (2 H, m), 7.19–7.22 (2 H, m),
7.27–7.35 (5 H, m). FABMS (positive-ion) m/z: 501
[M+H]+, 523 [M+Na]+. HRFABMS (positive-ion),

2084
M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
give a solid, which was chromatographed on a silica gel
column. Elution with 1:1 cyclohexane–EtOAc gave 10
(43.5 mg, 99%) as a solid; mp 164–166 °C (from
EtOAc–hexane). [h]2D4 −85.8° (c 0.4, CHCl3). IR
was stirred under hydrogen at rt for 30 min. After
filtration of the reaction mixture, the filtrate was con-
centrated in vacuo to give 11 (37 mg, quant) as a solid;
mp 188–190 °C (from EtOAc). [h]2D3 −64.5° (c 0.07,
MeOH). IR wmax(KBr) 3531, 3466, 3434, 3407, 3240,
wmax(KBr) 3410–2870, 1786, 1734 cm−1 1H NMR
.
(CDCl3): l 1.31 (3 H, s), 1.61 (3 H, s), 3.60 (1 H, dd, J
1.9, 10.5 Hz), 3.75 (1 H, dd, J 1.9, 10.5 Hz), 4.53, 4.63
(2 H, AB-q, J 11.4 Hz), 4.59 (1 H, m), 4.77 (1 H, d, J
5.7 Hz), 4.80 (1 H, d, J 5.7 Hz), 6.31 (1 H, bs, NH),
7.31–7.45 (5 H, m), 7.68 (1 H, bs, NH, exchanged on
addition of D2O). FABMS (positive-ion) m/z: 349
[M+H]+, 403 [M+Na+]. HRFABMS (positive-ion),
Calcd for C17H21N2O6: 349.1400; Found: 349.1388.
Anal. Calcd for C17H20N2O6: C, 58.61; H, 5.79; N, 8.04.
Found: C, 58.73; H, 5.77; N, 8.05.
2987, 2936, 1790, 1745, 1725 cm−1 1H NMR
.
(CD3OD): l 1.32 (3 H, s), 1.54 (3 H, s), 3.68 (1 H, dd,
J 3.4, 11.9 Hz), 3.71 (1 H, dd, J 3.3, 11.9 Hz), 4.46 (1
H, m), 4.81–4.93 (2 H, m). FABMS (positive-ion) m/z:
517 [2 M+H]+, 259 [M+H]+. HRFABMS (positive-
ion), Calcd for C10H15N2O6: 259.0930. Found:
259.0922.
(b) Compound 20 was treated as described in the
formation of 11 from 10 to give 11 (quant), which was
identical with that obtained from 10.
(b) To a solution of 14 (100 mg, 2.74 mmol) in
MeCN (3 mL) was added a solution of 1 M aq
NaHCO3 (2 mL) and aq 30% H2O2 (0.3 mL). After
stirring for 15 min at rt, the reaction mixture was
diluted with EtOAc, which was washed with 10% aq
NaHSO3 and brine. The organic extract was dried over
MgSO4 and filtered. The filtrate was concentrated in
vacuo to give 10 (96 mg, quant), which was identical
with that obtained from 9.
[5R - (5h,7i,8h,9h)] - 1,3 - Diaza - 8,9 - isopropylidenedi-
oxy-7-(hydroxymethyl)-6-oxaspiro[4.4]nonane-2,4-dione
(11%).—(a) Compound 11% was also obtained quantita-
tively as a powder from 10% according to the procedure
described above in the formation of 11 from 10. IR
wmax(KBr) 3700–2940, 1790, 1737 cm−1 1H NMR
.
(CD3OD): l 1.32 (3 H, s), 1.54 (3 H, s), 3.68 (1 H, dd,
J 3.4, 11.9 Hz), 3.71 (1 H, dd, J 3.3, 11.9 Hz), 4.46 (1
H, m), 4.81–4.93 (2 H, m). FABMS (positive-ion) m/z:
259 [M+H]+. HRFABMS (positive-ion), Calcd for
C10H15N2O6: 259.0930; Found: 259.0934.
(b) Compound 20% was treated as described in the
formation of 11 from 10 to give 11% (quant), which was
identical with that obtained from 10%.
[5R - (5h,7i,8h,9h)] - 7 - (Benzyloxymethyl) - 1,3 - diaza-
8,9-isopropylidenedioxy-6-oxaspiro[4.4]nonane-2,4-dione
(10%).—(a) Compound 10% was also obtained quantita-
tively as a powder from 9% using NH3 according to the
same procedure described in the above procedure (a) to
obtain 10 from 9. IR wmax(CHCl3) 3432, 1801, 1756,
[5S-(5h,7h,8i,9i)]-1,3-Diaza-8,9-dihydroxy-7-(hy-
droxymethyl)-6-oxaspiro[4.4]nonane-2,4-dione (12, hy-
dantocidin).—A solution of 11 (31 mg, 0.12 mmol) in
1:3 CF3COOH–water (5 mL) was stirred at 0 °C for 2
h, and concentrated in vacuo to give 12 (26 mg, quant)
as a solid, mp 190–192 °C (from MeOH–EtOAc). [h]D23
+28.3° (c 0.44, water) [lit. +29.0° (c 0.62, water)].3c IR
1
1385, 1091 cm−1. H NMR (CDCl3): l 1.36 (3 H, s),
1.57 (3 H, s), 3.61–3.68 (2 H, m), 4.40 (1 H, m), 4.57,
4.60 (2 H, AB-q, J 12.0 Hz), 4.76–4.82 (2 H, m), 6.05
(1 H, bs, NH), 7.25–7.40 (5 H, m), 8.00 (1 H, bs, NH,
exchanged on addition of D2O). FABMS (positive-ion)
m/z: 349 [M+H]+. HRFABMS (positive-ion), Calcd
for C17H21N2O6: 349.1400; Found: 349.1395.
wmax(KBr) 3470–2770, 1780, 1740, 1710 cm−1 1H
.
(b) To a solution of 9 (60 mg, 0.518 mmol) or 10 (60
mg) in DMF (4 mL) and THF (24 mL) was added
DBU (60 mg). The solution was stirred at rt for 7 days,
diluted with EtOAc, and washed with 0.01 M aq HCl,
water, and brine. The organic extract was dried over
MgSO4 and filtered, and the filtrate was evaporated in
vacuo to give a residue, which was chromatographed
on a silica gel column. Elution with 1:1 cyclohexane–
EtOAc gave an epimer 10% [50 mg (91%) from 9; or 52
mg (87%) from 10 accompanying the recovered starting
10 (7 mg, 12%)].
NMR (D2O, Me3SiCD2CD2COONa as an internal
standard): l 3.68 (1 H, dd, J 4.6, 12.7 Hz), 3.78 (1 H,
dd, J 3.0, 12.7 Hz), 4.22 (1 H, dd, J 4.4, 5.8 Hz), 4.34
(1 H, m), 4.40 (1 H, d, J 5.8 Hz). FABMS (positive-ion)
m/z: 219 [M+H]+. HRFABMS (positive-ion), Calcd
for C7H11N2O6: 219.0617; Found: 219.0623. Anal.
Calcd for C7H10N2O6: C, 38.53; H, 4.62; N, 12.84.
Found: C, 38.50; H, 4.73; N, 12.85.
[5R-(5h,7i,8h,9h)]-1,3-Diaza-8,9-dihydroxy-7-(hy-
droxymethyl)-6-oxaspiro[4.4]nonane-2,4-dione
(12%,
spiro-epi-hydantocidin).—Compound 12% was also ob-
tained from 11% as a powder according to the same
procedure described above. [h]2D3 −9.8° (c 0.75, MeOH)
[lit. −11.0° (c 0.3, MeOH)].3j IR wmax(KBr) 3470–2770,
(c) Compound 10% was also obtained quantitatively
from 14% according to the procedure described above to
obtain 10 from 14. This compound was identical with
that obtained above from 9%.
1
1780, 1740, 1710 cm−1. H NMR (CD3OD), l 3.59 (1
[5S-(5h,7h,8i,9i)]-1,3-Diaza-7-(hydroxymethyl)-8,9-
isopropylidenedioxy - 6 - oxaspiro[4.4]nonane - 2,4 - dione
(11).—(a) A solution of 10 (50 mg, 0.14 mmol) in
EtOAc (10 mL) containing 10% Pd-on-charcoal (25 mg)
H, dd, J 5.2, 12.2 Hz, C-6ꢀH), 3.78 (1 H, dd, J 4.3, 12.2
Hz, C-6ꢀH), 4.08 (1 H, ddd, J 3.4, 4.3, 5.2 Hz, C-5ꢀH),
4.16 (1 H, dd, J 3.4, 5.0 Hz, C-4ꢀH), 4.24 (1 H, d, J 5.0
Hz, C-3ꢀH). FABMS (positive-ion) m/z: 219 [M+H]+.

M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
2085
HRFABMS (positive-ion), Calcd for C7H11N2O6:
219.0617; Found: 219.0598. Anal. Calcd for
C7H10N2O6: C, 38.54; H, 4.62; N, 12.84. Found: C,
38.33; H, 4.86; N, 12.70.
(113 mg, 19.6%; Rf 0.449 (2:1 cyclohexane–EtOAc))
and 14 (466 mg, 80.5%; Rf 0.359 (2:1 cyclohexane–
EtOAc)). Physical data of 14: [h]2D4 −169.7° (c 3.7,
CHCl3). IR wmax(KBr) 3400–2860, 1781, 1498 cm−1
.
Methyl
cyanato-i-
6-O-benzyl-3,4-O-isopropylidene-2-isothio-
-ribo-2-hexulofuranosonate (13) and Methyl
1H NMR (CDCl3): l 1.30 (3 H, s), 1.61 (3 H, s), 3.62 (1
H, dd, J 1.5, 10.3 Hz), 3.78 (1 H, dd, J 1.5, 10.3 Hz),
4.56, 4.64 (2 H, AB-q, J 11.4 Hz), 4.61 (1 H, t, J 1.5
Hz), 4.78 (1 H, d, J 5.9 Hz), 4.81 (1 H, d, J 5.9 Hz),
7.38–7.47 (5 H, m), 7.78 (1 H, s, NH, exchanged on
addition of D2O), 8.16 (1 H, s, NH, exchanged on
addition of D2O). FABMS (positive-ion) m/z: 365
[M+H]+. HRFABMS (positive-ion), Calcd for
C17H21N2O5S: 365.1171; Found: 365.1167. Anal. Calcd
for C17H20N2O5S: C, 56.03; H, 5.53; N, 7.69; S, 8.80.
Found: C, 55.57; H, 5.67; N, 7.44; S, 8.56. Physical
data of 14%: [h]D24 −61.5° (c 1.3, CHCl3). IR wmax(KBr)
D
6-O-benzyl-3,4-O-isopropylidene-2-isothiocyanato-h-D-
ribo-2-hexulofuranosonate (13%).—To a solution of 4
(3.57 g, 10.00 mmol) in DMF (36 mL) was added
KSCN (3.00 g, 30.87 mmol). After 5 h stirring at 80 °C,
the reaction mixture was diluted with EtOAc, and
washed with water and brine. The organic extract was
dried over MgSO4, filtered, and concentrated in vacuo
(finally with a pump) to give a residue, which was
chromatographed on a silica gel column. Elution with
2:1 cyclohexane–EtOAc gave a 4:1 mixture of 13 and
13% (3.26 g, 84%). The mixture was partially separated
on a TLC plate by development with 3:1 cyclohexane–
EtOAc (13, Rf 0.500; 13%, Rf 0.467) to measure the
physical data. Physical data of 13: [h]2D4 −118.8° (c 3.9,
CHCl3). IR wmax(film) 3090–2860, 2016, 1773, 1756
3450–2850, 1755, 1505 cm−1 1H NMR (CDCl3): l
.
1.36 (3 H, s), 1.59 (3 H, s), 3.62 (1 H, dd, J 6.2, 10.4
Hz), 3.67 (1 H, dd, J 6.2, 10.4 Hz), 4.43 (1 H, t, J 6.2
Hz), 4.59 (2 H, s), 4.81 (2 H, s), 7.26 (1 H, bs, NH,
exchanged on addition of D2O), 7.28–7.36 (5 H, m),
8.39 (1 H, bs, NH, exchanged on addition of D2O).
FABMS (positive-ion) m/z: 365 [M+H]+. HRFABMS
(positive-ion), Calcd for C17H21N2O5S: 365.1171;
Found: 365.1183. Anal. Calcd for C17H20N2O5S: C,
56.03; H, 5.53; N, 7.69; S, 8.80. Found: C, 56.43; H,
5.46; N, 7.44; S, 8.50.
1
cm−1. H NMR (CDCl3): l 1.30 (3 H, s), 1.44 (3 H, s),
3.60–3.68 (2 H, m), 3.89 (3 H, s), 4.55, 4.65 (2 H, AB-q,
J 11.7 Hz), 4.64 (1 H, m), 4.89–4.92 (2 H, m), 7.30–
7.37 (5 H, m). 13C NMR (CDCl3): l 25.0 (CH3), 25.8
(CH3), 53.4 (OCH3), 69.6 (CH2), 73.8 (CH2), 82.1 (CH),
86.9 (CH), 89.2 (CH), 98.1 (anomeric C), 114.2
(O2CMe2), 127.8 (aromatic CH×2), 127.9 (aromatic
CH), 128.4 (aromatic CH×2), 137.5 (aromatic C),
143.2 (ꢀNCS), 165.0 (COOMe). FABMS (positive-ion)
m/z: 402 [M+Na]+, 380 [M+H]+. Anal. Calcd for
C18H21NO6S: C, 56.98; H, 5.58; N, 3.69; S, 8.45.
Found: C, 57.19; H, 5.57; N, 3.17; S, 8.17. Physical
data of 13%: [h]D24 −36.3° (c 0.2, CHCl3). 1H NMR
(CDCl3): l 1.39 (3 H, s), 1.69 (3 H, s), 3.63–3.70 (2 H,
m), 3.79 (3 H, s), 4.50 (1 H, m), 4.51, 4.57 (2 H, AB-q,
J 12.0 Hz), 4.78 (1 H, m), 5.03 (1 H, d, J 6.6 Hz),
7.30–7.42 (5 H, m). 13C NMR (CDCl3): l 25.3 (CH3),
26.1 (CH3), 53.6 (OCH3), 69.5 (CH2), 73.5 (CH2), 81.1
(CH), 83.7 (CH), 84.3 (CH), 95.3 (anomeric C), 116.4
(O2CMe2), 127.6 (aromatic CH×2), 128.4 (aromatic
CH×2), 137.5 (aromatic C), 144.4 (ꢀNCS), 166.2
(COOMe). FABMS (positive-ion) m/z: 402 [M+Na]+,
380 [M+H]+. HRFABMS (positive-ion), Calcd for
C18H22NO6S: 380.1168; Found: 380.1169.
2,3-O-Isopropylidene-5-O-(4-methoxybenzyl)-D-ri-
bono-1,4-lactone (15).—Compound 15 (10.37 g, 77%)
was also obtained as an oil from 1 (8.2 g, 43.57 mmol),
4-methoxybenzyl chloride (8.20 g, 52.36 mmol) and
NaH (60–70% oil dispersion, 2.09 g) in DMF (30 mL)
according to the same procedure described in the for-
mation of 2 from 1. IR wmax(film) 3030–2840, 1785,
1614, 1587, 1515 cm−1. 1H NMR (CDCl3): l 1.36 (3 H,
s), 1.47 (3 H, s), 3.64 (1 H, dd, J ꢀ1.0, 10.3 Hz), 3.68
(1 H, dd, J 1.8, 10.3 Hz), 3.81 (3 H, s), 4.40, 4.48 (2 H,
AB-q, J 11.7 Hz), 4.63 (1 H, dd, J 1.0, 1.8 Hz), 4.68 (1
H, d, J 5.5 Hz), 4.76 (1 H, d, J 5.5 Hz), 6.88 (2 H, d,
J 8.4 Hz), 7.18 (2 H, d, J 8.4 Hz). FABMS (positive-
ion) m/z: 308 [M+], 309 [M+H]+, 331 [M+Na]+
HRFABMS (positive-ion), Calcd for C16H20O6:
308.1260; Found: 308.1263. Anal. Calcd for C16H20O6:
C, 62.33; H, 6.54. Found: C, 61.98; H, 6.75.
2,5-Anhydro-1,1-dichloro-1-deoxy-3,4-O-isopropyli-
[5S - (5h,7h,8i,9i)] - 7 - (Benzyloxymethyl) - 1,3 - diaza-
8,9 - isopropylidenedioxy - 6 - oxa - 2 - thioxo - spiro[4.4]no-
nan-4-one (14) and [5R-(5h,7i,8h,9h)]-7-(benzy-
loxymethyl)-1,3-diaza-8,9-isopropylidenedioxy-6-oxa-
2-thioxo-spiro[4.4]nonan-4-one (14%).—To a solution of
a 4:1 mixture of 13 and 13% (600 mg, 1.58 mmol) in
MeOH (6.0 mL) was added a solution of 2 M NH3 in
MeOH (2.0 mL) at 24 °C with stirring. After 2 h the
reaction mixture was concentrated in vacuo to give a
solid, which was chromatographed on a silica gel
column. Elution with 3:1 cyclohexane–EtOAc gave 14%
dene - 6 - O - (4 - methoxybenzyl) - D - ribo - hex - 1 - enitol
(16).—Compound 15 (4.77 g) was treated as described
in the formation of 3 from 2 to give 16 (5.54 g, 95%) as
a
solid, mp 62–65 °C (thread-like needles from
EtOAc–hexane). [h]2D4 −147.3° (c 1.1, CHCl3). IR
wmax(KBr) 3040–2830, 1733, 1664, 1610, 1585, 1513
cm−1. H NMR (CDCl3): l 1.38 3 H, s), 1.48 (3 H, s),
1
3.59 (1 H, dd, J 2.9, 11.0 Hz), 3.63 (1 H, dd, J 2.9, 11.0
Hz), 3.82 (3 H, s), 4.42, 4.45 (2 H, AB-q, J 11.7 Hz),
4.64 (1 H, t, J 2.9 Hz), 4.77 (1 H, d, J 5.9 Hz), 5.23 (1
H, d, J 5.9 Hz), 6.88–6.90 (2 H, m), 7.18–7.21 (2 H,

2086
M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
m). FABMS (positive-ion) m/z: 374 [M+]. HRFABMS
(positive-ion), Calcd for C17H20 35Cl2O5: 374.0688;
Found: 374.0691. Anal. Calcd for C17H20Cl2O5: C,
54.41; H, 5.37; Cl, 18.90. Found: C, 54.30; H, 5.38; Cl,
18.66.
oxa-2-thioxo-1,3-diazaspiro[4.4]nonan-4-one (19%).—A
5:1 mixture of 18 and 18% (130 mg, 0.317 mmol) was
treated as described in the formation of 14 and 14% from
a mixture of 13 and 13% to give 19% (20 mg, 16%, Rf
0.474 in 3:2 cyclohexane–EtOAc) and 19 (105 mg, 84%,
Rf 0.382 in 3:2 cyclohexane–EtOAc). Physical data of
19%: [h]2D4 −58.1° (c 0.32, CHCl3). IR wmax(film) 3226,
Methyl 2,5-anhydro-2-chloro-3,4-O-isopropylidene-6-
O-(4-methoxybenzyl)-h-
(17) and methyl 2,5-anhydro-2-chloro-3,4-O-isopropyli-
dene-6-O-(4-methoxybenzyl)-i- -ribo-2-hexulofurano-
D -ribo-2-hexulofuranosonate
2993–2838, 1768, 1613, 1587, 1513, 1465 cm−1 1H
.
D
NMR (CDCl3): l 1.36 (3 H, s), 1.59 (3 H, s), 3.58–3.65
(2 H, m), 3.80 (3 H, s), 4.41 (1 H, m), 4.52 (2 H, s),
4.77–4.82 (2 H, m), 6.88 (2 H, d, J 8.8 Hz), 7.28 (2 H,
d, J 8.8 Hz), 7.40 (1 H, bs, NH), 8.79 (1 H, broad,
NH). FABMS (positive-ion) m/z: 395 [M+H]+. HR-
FABMS (positive-ion), Calcd for C18H23N2O6S:
395.1276. Found: 395.1274. Physical data of 19: [h]D24
−154.4° (c 1.3, CHCl3). IR wmax(film) 3392, 3225,
sonate (17%).—Compound 16 (3.752 g) was treated as
described in the formation of 4 from 3 to give 17% (507
mg, 13%) and 17 (2.07 g, 54%). Physical data of 17%: IR
1
wmax(film) 3000–2830, 1766, 1613, 1587, 1514 cm−1. H
NMR (CDCl3): l 1.33 (3 H, s), 1.43 (3 H, s), 3.71 (2 H,
d, J 7.2 Hz),), 3.81 (3 H, s), 3.89 (3 H, s), 4.50, 4.53 (2
H, AB-q, J 11.0 Hz), 4.63 (1 H, dt, J 1.6, 7.2 Hz), 4.92
(1 H, dd, J 1.8, 5.6 Hz, C-4ꢀH), 5.13 (1 H, d, J 5.6 Hz),
6.88 (2 H, d, J 8.8 Hz), 7.27 (2 H, d, J 8.8 Hz). FABMS
(positive-ion) 386 [35Cl, M+]. HRFABMS (positive-
ion), Calcd for C18H23 35ClO7: 386.1123; Found:
386.1125. Physical data of 17: IR wmax(film) 2990–2860,
2986–2838, 1781, 1613, 1586, 1513, 1500 cm−1 1H
.
NMR (CDCl3): l 1.30 (3 H, s), 1.61 (3 H, s), 3.60 (1 H,
dd J 1.8, 10.6 Hz), 3.75 (1 H, dd J 1.8, 10.6 Hz), 3.84
(3 H, s), 4.50, 4.56 (2 H, AB-q, J 11.0 Hz), 4.61 (1 H,
m), 4.76 (1 H, d, J 5.5 Hz), 4.80 (1 H, d, J 5.5 Hz), 6.96
(2 H, d, J 8.8 Hz), 7.31 (2 H, d, J 8.8 Hz), 7.80 (1 H,
s, NH), 8.53 (1 H, broad, NH). FABMS (positive-ion)
m/z: 395 [M+H]+. HRFABMS (positive-ion), Calcd
for C18H23N2O6S: 395.1276; Found: 395.1279. Anal.
Calcd for C18H22N2O6S: C, 54.81; H, 5.62; N, 7.10; S,
8.13. Found: C, 54.54; H, 5.61; N, 7.01; S, 8.06.
[5S - (5h,7h,8i,9i)] - 8,9 - Isopropylidenedioxy - 7 - [(4-
methoxybenzyl)oxymethyl] - 6 - oxa - 1,3 - diazaspiro[4.4]-
nonane-2,4-dione (20).—Compound 19 was treated as
described in the formation of 10 from 14 to give 20
(96% after chromatographic purification) as a solid, mp
180–181 °C (from EtOAc). IR wmax(KBr) 3500–2830,
1793, 1740, 1612 cm−1. 1H NMR (CDCl3): l 1.30 (3 H,
s), 1.61 (3 H, s), 3.57 (1 H, dd, J 1.8, 10.6 Hz), 3.71 (1
H, dd, J 1.8, 10.6 Hz), 3.83 (3 H, s), 4.47, 4.54 (2 H,
AB-q, J 11.0 Hz), 4.57 (1 H, m), 4.75 (1 H, d, J 5.9 Hz),
4.77 (1 H, d, J 5.9 Hz), 6.36 (1 H, s, NH), 6.94 (2 H, d,
J 8.8 Hz), 7.26 (2 H, d, J 8.8 Hz), 7.86 (1 H, bs, NH).
FABMS (positive-ion) m/z: 379 [M+H]+. HRFABMS
(positive-ion), Calcd for C18H23N2O7: 379.1506; Found:
379.1508.
1762, 1751, 1613, 1587, 1514 cm−1 1H NMR (500
.
MHz, CDCl3): l 1.39 (3 H, s), 1.70 (3 H, s), 3.63–3.70
(2 H, m), 3.81 (3 H, s), 3.83 (3 H, s), 4.45, 4.51 (2 H,
AB-q, J 11.7 Hz), 4.57 (1 H, dd, J 2.9, 5.8 Hz), 4.78 (1
H, dd, J 2.9, 6.8 Hz, C-4ꢀH), 5.00 (1 H, d, J 7.8 Hz),
6.88 (2 H, d, J 8.8 Hz), 7.22 (2 H, d, J 8.8 Hz). FABMS
(positive-ion) m/z: 386 [35Cl, M+], 387 [M+H]+, 425
[M+K]+ (on addition of KI). HRFABMS (positive-
ion), Calcd for C18H23 35ClO7 K: 425.0770; Found:
425.0779.
Methyl 2,5-anhydro-3,4-O-isopropylidene-2-isothio-
cyanato-6-O-(4-methoxybenzyl)-h-
anosonate (18) and methyl 2,5-anhydro-3,4-O-isopropy-
lidene-2-isothiocyanato-6-O-(4-methoxybenzyl)-i-
D-ribo-2-hexulofur-
D-
ribo-2-hexulofuranosonate (18%).—Compound 17 (193
mg) was treated as described in the formation of 13 and
13% from 4 to give a chromatographically inseparable
5:1 mixture of 18 and 18% (149 mg, 73%). IR wmax(film)
3000–2830, 2017, 1772, 1756, 1613, 1587, 1514 cm−1
.
Anal. Calcd for C19H23NO7S (409.4): C, 55.73; H, 5.66;
N, 3.42; S, 7.83. Found: C, 55.29; H, 5.78; N, 3.24; S,
1
7.70. H NMR (CDCl3) of 18: l 1.29 (3 H, s), 1.43 (3
[5R - (5h,7i,8h,9h)] - 8,9 - Isopropylidenedioxy - 7 - [(4-
methoxybenzyl)oxymethyl] - 6 - oxa - 1,3 - diazaspiro[4.4]-
nonane-2,4-dione (20%).—Compound 19% was treated as
described in the formation of 10 from 14 to give 20%
(95% after chromatographic purification) as an amor-
phous solid. IR wmax(film) 3259 (broad), 1793, 1740,
H, s), 3.57–3.63 (2 H, m), 3.82 (3 H, s), 3.88 (3 H, s),
4.49, 4.57 (2 H, AB-q, J 11.7 Hz), 4.63 (1 H, dt, J 1.0,
4.8 Hz), 4.87 (1 H, dd, J 1.0, 5.7 Hz), 4.89 (1 H, d, J 5.7
1
Hz), 6.89 (2 H, d, J 8.7 Hz), 7.28 (2 H, d, J 8.7 Hz). H
NMR (CDCl3) of 18%: l 1.38 (3 H, s), 1.68 (3 H, s),
3.57–3.63 (2 H, m), 3.81 (3 H, s), 3.82 (3 H, s), 4.43,
4.50 (2 H, AB-q, J 11.7 Hz), 4.48 (1 H, m), 4.75 (1 H,
dd, J 2.5, 6.6 Hz), 5.02 (1 H, d, J 6.6 Hz), 6.88 (2 H, d,
J 8.7 Hz), 7.21 (2 H, d, J 8.7 Hz).
[5S - (5h,7h,8i,9i)] - 8,9 - Isopropylidenedioxy - 7 - [4-
(methoxbenzyloxy)methyl] - 6 - oxa - 2 - thioxo - 1,3 - diaza-
spiro[4.4]nonan-4-one (19) and [5R-(5h,7i,8h,9h)]-8,9-
isopropylidenedioxy-7-[4-(methoxbenzyloxy)methyl]-6-
1
1613 cm−1. H NMR (CDCl3): l 1.35 (3 H, s), 1.56 (3
H, s), 3.60 (1 H, dd, J 6.5, 10.3 Hz), 3.62 (1 H, dd, J
6.4, 10.3 Hz), 3.80 (3 H, s), 4.37 (1 H, dt, J 1.4, 6.5 Hz),
4.49, 4.52 (2 H, AB-q, J 11.7 Hz), 4.77 (2 H, singlet-
like), 6.38 (1 H, s, NH), 6.88 (2 H, d, J 8.8 Hz), 7.27 (2
H, d, J 8.8 Hz), 8.85 (1 H, bs, NH). FABMS (positive-
ion) m/z: 378 [M+]. HRFABMS (positive-ion), Calcd
for C18H22N2O7: 378.1427. Found: 378.1424.

M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
2087
Methyl 6-O-acetyl-2,5-anhydro-2-chloro-3,4-O-iso-
propylidene-h- -ribo-2-hexulofuranosonate (21).—A
one (23).—(a) To a solution of 22 (18 mg, 0.054 mmol)
in MeOH (1 mL) was added a solution of 2 M NH3 in
MeOH (0.5 mL). The solution was stirred for 6 h at rt,
and concentrated in vacuo to give thioxohydantoine 23
quantitatively as a solid, mp 158–161 °C (from CHCl3).
[h]2D4 −174.9° (c 0.3, MeOH). IR wmax(KBr) 3500–
D
solution of 4 (350 mg, 0.981 mmol) in AcOEt (25 mL)
was hydrogenolyzed at rt for 20 min using 10% Pd-on-
charcoal (50 mg) as a catalyst. The catalyst was filtered,
and to this filtrate was added Ac2O (2 mL) and pyridine
(1 mL). This solution was stirred for 15 h at rt and
concentrated in vacuo to give a residue, which was
chromatographed on a silica gel column. Elution with
1:1 cyclohexane–EtOAc gave 21 (273 mg, 90%) as a
solid, mp 105–107 °C (from EtOAc–hexane). IR
1
3000, 1763, 1509 cm−1. H NMR (CDCl3+D2O): l
1.34 (3 H, s), 1.64 (3 H, s), 3.87 (1 H, dd, J 1.8, 11.4
Hz), 3.93 (1 H, dd, J 1.8, 11.4 Hz), 4.63 (1 H, s), 4.92
(1 H, d, J 5.9 Hz), 4.95 (1 H, d, J 5.9 Hz). FABMS
(positive-ion) m/z: 275 [M+H]+. HRFABMS (posi-
tive-ion), Calcd for C10H15N2O5S: 275.0702; Found:
275.0705. Anal. Calcd for C10H14N2O5S: C, 43.79; H,
5.14; N, 10.21; S, 11.69. Found: C, 43.67; H, 5.01; N,
10.23; S, 11.87.
1
wmax(film) 3000–2950, 1743 cm−1. H NMR (CDCl3): l
1.41 (3 H, s), 1.71 (3 H, s), 2.10 (3 H, s), 3.89 (3 H, s),
4.29 (1 H, dd, J 4.9, 12.2 Hz), 4.38 (1 H, dd, J 3.9, 12.2
Hz), 4.64 (1 H, dd, J 3.9, 7.8 Hz), 4.71 (1 H, dd, J 3.9,
6.8 Hz), 5.03 (1 H, d, J 7.8 Hz). Anal. Calcd for
C12H17ClO7: C, 46.69; H, 5.55; Cl, 11.48. Found: C,
46.78; H, 5.53; Cl, 11.39.
(b) To a solution of 19 (64 mg, 0.160 mmol) and
PhSH (45 mg, 0.408 mmol) in CH2Cl2 (4 mL) was
added SnCl4 (1 M solution in CH2Cl2, 0.40 mL) at
−78 °C under nitrogen. After stirring for 30 min at
−78 °C, the reaction mixture was diluted with EtOAc,
and the solution was washed with satd aq NaHCO3 and
brine. The organic extract was dried over MgSO4, and
filtered. The filtrate was concentrated in vacuo to give a
residue, which was chromatographed on a silica gel
column. Elution with 2:1 cyclohexane–EtOAc gave the
starting 19 (4 mg, 6% recovery) and 23 (36 mg, 81%),
which was identical with that obtained from 22.
[5R-(5h,7i,8h,9h)]-1,3-Diaza-7-hydroxymethyl-8,9-
isopropylidenedioxy-6-oxa-2-thioxo-spiro[4.4]nonan-4-
one (23%).—(a) Compound 23% was also obtained in 72%
yield as a powder from 22% according to the procedure
described above to obtain 23 from 22. [h]2D4 −43.6° (c
0.3, MeOH). IR wmax(KBr) 4215, 3420, 3000–2870,
Methyl 6-O-acetyl-2,5-anhydro-3,4-O-isopropylidene-
2-isothiocyanato-i-
and methyl 6-O-acetyl-2,5-anhydro-3,4-O-isopropylidene
- 2 - isothiocyanato - h - - ribo - 2 - hexulofuranosonate
D-ribo-2-hexulofuranosonate
(22)
D
(22%).—To a solution of 21 in DMF (2 mL) was added
KSCN (94 mg, 0.967 mmol). After 15 h stirring at
80 °C, the reaction mixture was diluted with EtOAc and
washed with water and brine. The organic extract was
dried over MgSO4, filtered, and concentrated in vacuo
(finally with a pump) to give a residue, which was
chromatographed on a silica gel column. Elution with
2:1 cyclohexane–EtOAc gave 22 (73 mg, 68%, Rf 0.547)
and 22% (14 mg, 13%, Rf 0.453). Physical data of 22: IR
wmax(film) 2990–2950, 2011, 1773, 1749 cm−1. 1H NMR
(CDCl3): l 1.33 (3 H, s), 1.46 (3 H, s), 2.14 (3 H, s), 3.91
(3 H, s), 4.20 (1 H, dd, J 5.9, 12.1 Hz), 4.31 (1 H, dd, J
5.9, 12.1 Hz), 4.68 (1 H, ddd, J 1.5, 5.1, 5.9 Hz), 4.86 (1
H, dd, J 1.5, 5.9 Hz), 4.94 (1 H, d, J 5.1 Hz). 13C NMR
(CDCl3): l 20.8 (CH3), 25.1 (CH3), 25.9 (CH3), 53.6
(OCH3), 63.4 (CH2), 81.9 (CH), 85.8 (CH), 89.0 (CH),
97.7 (anomeric C), 114.7 (O2CMe2), 144.5 (ꢀNCS),
164.7 (COOMe), 170.4 (MeCOO). FABMS (positive-
ion) m/z: 332 [M+H]+, 354 [M+Na]+. Anal. Calcd
for C13H17NO7S: C, 47.12; H, 5.17; N, 4.23; S, 9.68.
Found: C, 47.27; H, 5.15; N, 4.21; S, 9.33. Physical data
1
1760 cm−1. H NMR (CDCl3+D2O): l 1.39 (3 H, s),
1.62 (3 H, s), 3.71 (1 H, dd, J 2.9, 13.2 Hz, on addition
of D2O), 3.87 (1 H, dd, J 2.2, 13.2 Hz), 4.08 (1 H, bs,
OH), 4.51 (1 H, m), 4.92 (1 H, d, J 5.9 Hz), 5.04 (1 H,
dd, J 1.5, 5.9 Hz), 7.31 (1 H, bs, HH), 9.23 (1 H, bs,
NH). FABMS (positive-ion) m/z: 275 [M+H]+. HR-
FABMS (positive-ion), Calcd for C10H15N2O5S:
275.0702; Found: 275.0707. Anal. Calcd for
C10H14N2O5S: C, 43.79; H, 5.14; N, 10.21; S, 11.69.
Found: C, 43.46; H, 4.91; N, 10.49; S, 12.17.
(b) Compound 19% was treated as described in the
formation of 23 from 19 to give starting 19% (13%
recovery) and 23% (76%), which was identical with that
obtained from 22%.
[5S-(5h,7h,8i,9i)]-1,3-Diaza-7-hydroxymethyl-8,9-
dioxy-6-oxa-2-thioxo-spiro[4.4]nonan-4-one (24, C-2-
thioxo-hydantocidin) and its spiro-epimer (24%).—A solu-
tion of 23 (18 mg, 0.066 mmol) in 1:3 CF3COOH–water
(0.4 mL) was stirred at 0 °C for 2 h, then −5 °C for 16
h. The solution was concentrated in vacuo to give a
mixture, which was chromatographed on a preparative
silica gel plate. Development with 9:1 EtOAc–MeOH
gave two bands of products. The products from each
1
of 22%: IR wmax(film) 2990–2950, 2022, 1750 cm−1. H
NMR (CDCl3): l 1.40 (3 H, s), 1.70 (3 H, s), 2.12 (3 H,
s), 3.89 (3 H, s), 4.18 (1 H, dd, J 4.8, 12.1 Hz), 4.37 (1 H,
dd, J 3.7, 12.1 Hz), 4.54 (1 H, m), 4.71 (1 H, dd, J 2.8,
6.9 Hz), 5.02 (1 H, d, J 7.3 Hz). 13C NMR (CDCl3): l
20.8 (CH3), 25.4 (CH3), 26.1 (CH3), 53.9 (OCH3), 63.2
(CH2), 80.9 (CH), 82.5 (CH), 84.3 (CH), 94.9 (anomeric
C), 117.3 (O2CMe2), 145.0 (ꢀNCS), 166.0 (COOMe),
170.4 (MeCOO). FABMS (positive-ion) m/z: 332 [M+
H]+. HRFABMS (positive-ion), Calcd for C13H18NO7S:
332.0804; Found: 332.0797.
[5S-(5h,7h,8i,9i)]-1,3-Diaza-7-hydroxymethyl-8,9-
isopropylidenedioxy-6-oxa-2-thioxo-spiro[4.4]nonan-4-

2088
M. Shiozaki / Carbohydrate Research 337 (2002) 2077–2088
(d) Fonne-Pfister, R.; Chemla, P.; Ward, E.; Girardet,
M.; Kreuz, K. E.; Honzatko, R. B.; Fromm, H. J.; Scha¨r,
H-P.; Gru¨tter, M. G.; Cowan-Jacob, S. W. Proc. Natl.
Acad. Sci. USA 1996, 93, 9431–9436.
band were eluted with 9:1 EtOAc–MeOH, and the
eluants were diluted with EtOAc. Each diluted solution
was washed with brine. The organic extract was dried
over MgSO4, filtered, and concentrated in vacuo to give
24 (9.4 mg, 61%, Rf 0.400) as a powder, and also 24%
(2.4 mg, 16%, Rf 0.433, identical with that obtained
3. (a) Mio, S.; Ichinose, R.; Goto, K.; Sugai, S.; Sato, S.
Tetrahedron 1991, 47, 2111–2120;
(b) Mio, S.; Ichinose, R.; Shiraishi, M.; Sugai, S.;
Haruyama, H.; Sato, S. Tetrahedron 1991, 47, 2121–
2132;
(c) Mio, S.; Kumagawa, Y.; Sugai, S. Tetrahedron 1991,
47, 2133–2144;
(d) Fairbanks, A. J.; Ford, P. S.; Watkin, D. J.; Fleet, G.
W. J. Tetrahedron Lett. 1993, 34, 3327–3330;
(e) Burton, J. W.; Son, J. C.; Fairbanks, A. J.; Choi, S.
S.; Taylor, H.; Watkin, D. J.; Winchester, B. G.; Fleet,
G. W. J. Tetrahedron Lett. 1993, 34, 6119–6122;
(f) Chemla, P. Tetrahedron Lett. 1993, 34, 7391–7394;
(g) Harrington, P. M.; Jung, M. E. Tetrahedron Lett.
1994, 35, 5145–5148;
(h) Fairbanks, A.; Fleet, G. W. J. Tetrahedron 1995, 51,
3881–3894;
(i) Nakajima, N.; Matsumoto, M.; Katoh, T.; Terashima,
S. Tetrahedron 1996, 52, 1177–1194;
(j) Mio, S.; Ueda, M.; Hamura, M.; Kitagawa, J.; Sugai,
S. Tetrahedron 1991, 47, 2145–2154;
(k) Sano, H.; Mio, S.; Kitagawa, J.; Shindou, M.;
Honma, T.; Sugai, S. Tetrahedron 1995, 51, 12563–
12572.
from 23%), respectively. Physical data of 24: [h]D24
−
41.0° (c 0.4, MeOH) (lit.3k [h]D24 −52.7° (c 0.8,
MeOH)). IR wmax(KBr) 3329 (broad), 1760, 1676, 1516
1
cm−1. H NMR (CD3OD): l 3.62 (1 H, dd, J 4.1, 12.1
Hz), 3.65 (1 H, dd, J 3.8, 12.1 Hz), 4.07 (1 H, dd, J 2.6,
5.9 Hz), 4.25 (1 H, m), 4.30 (1 H, dd, J 1.4, 5.9 Hz).
FABMS (positive-ion) m/z: 235 [M+H]+. Anal. Calcd
for C7H10N2O5S·1.2 H2O: C, 32.86; H, 4.89; N, 10.95;
S, 12.58. Found: C, 33.11; H, 5.01; N, 10.61; S, 12.23.
[5R-(5h,7i,8h,9h)]-1,3-Diaza-7-hydroxymethyl-8,9-
dioxy-6-oxa-2-thioxo-spiro[4.4]nonan-4-one (24%).—A
solution of 23% (25 mg, 0.09 mmol) in 1:3 CF3COOH–
water (2.5 mL) was stirred at 20 °C for 2 h. The
solution was concentrated in vacuo to give a residue,
which was chromatographed on a silica gel column.
Elution with EtOAc gave 24% (20 mg, 94%) as a powder.
[h]D24 +35.0° (c 0.4, MeOH) (lit.3k [h]2D4 +36.8° (c 0.9,
MeOH)). IR wmax(KBr) 3334 (broad), 2930, 1763, 1630
4. Shiozaki, M. Carbohydr. Res. 2001, 335, 147–150.
5. Shiozaki, M.; Arai, M.; Kobayashi, Y.; Kasuya, A.;
Miyamoto, S. J. Org. Chem. 1994, 59, 4450–4460.
6. Lakhrissi, M.; Chapleur, Y. Tetrahedron Lett. 1998, 39,
4659–4662.
7. (a) Wheeler, T. N. J. Org. Chem. 1984, 49, 706–709;
(b) Speziale, A. J.; Marco, G. J.; Ratts, K. W. J. Am.
Chem. Soc. 1960, 82, 1260;
1
(w), 1510 cm−1. H NMR (CD3OD): l 3.60 (1 H, dd,
J 5.1, 12.4 Hz), 3.66 (1 H, dd, J 4.0, 12.4 Hz), 4.13 (1
H, m), 4.18 (1 H, dd, J 3.7, 4.4 Hz), 4.28 (1 H, d, J 4.4
Hz). FABMS (positive-ion) m/z: 235 [M+H]+. HR-
FABMS (positive-ion), Calcd for C7H11N2O5S:
235.0389; Found: 235.0381.
(c) Ravinowitz, R.; Marcus, R. J. Am. Chem. Soc. 1962,
84, 1312–1313;
(d) Chapleur, Y. J. Chem. Soc., Chem. Commun. 1984,
449–450;
(e) Bandzouzi, A.; Chapleur, Y. J. Chem. Soc., Perkin
Trans. 1 1987, 661–664;
(f) Lakhrissi, M.; Chapleur, Y. J. Org. Chem. 1994, 59,
5752–5757.
Acknowledgements
I would like to thank Dr. Shigeru Mio for helpful
advice and discussions.
8. (a) Kamarasa, M. J.; Ferna´ndez-Resa, P.; Garc´ıa-Ro´pez,
M. T.; De Las Heras, F. G.; Me´ndez-Castrillo´n, P. P.;
San Felix, A. Synthesis 1984, 509–510;
(b) Gasch, C.; Salameh, B. A. B.; Pradera, M. A.;
Fuentes, J. Tetrahedron Lett. 2001, 42, 8615–8617.
9. (a) Lindhorst, T. K.; Kieburg, K. Synthesis 1995, 1228–
1230;
(b) Somsa´k, L.; Czifra´k, K.; Deim, T.; Szila´gyi, L.;
Be´nyei, A. Tetrahedron: Asymmetry 2001, 12, 731–736.
10. (a) Yao, H.; Richardson, D. E. J. Am. Chem. Soc. 2000,
122, 3220–3221;
(b) P-Santander, C.; Scott, A. I. Tetrahedron Lett. 2000,
41, 2825–2829.
11. Mio, S.; Sano, H. Annu. Rep. Sankyo Res. Lab. 1997, 49,
91–119.
12. Yu, W.; Su, M.; Gao, X.; Yang, Z.; Jin, Z. Tetrahedron
Lett. 2000, 41, 4015–4017.
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