Synthesis of enantiomers of indanoxazolidinone based on the lipase-catalyzed resolution of the corresponding N-carbamylamino derivative

Article abstract of DOI:10.1016/S0040-4020(01)00454-9

Enantiomerically enriched (4R,5S)- and (4S,5R)-indano[1,2-d]oxazolidinones were enzymatically prepared from (±)-1-amino-2-indanol. Racemic 1-(N′-chloroacetyl-N-carbamylamino)-2-indanol O-chloroacetate was hydrolyzed with immobilized Pseudomonas cepacia lipase in the presence of β-cyclodextrin in acetone-buffer solution, to afford (1S,2R)-1-(N′-chloroacetyl-N-carbamylamino)-2-indanol (90%e.e.) and the unreacted (1R,2S)-substrate (97%e.e.), in nearly quantitative yields. The deprotection provided enantiomers of 1-N-carbamylamino-2-indanol, the precursor of indanoxazolidinone, via nitrosation-deaminocyclization reaction.

Full text of DOI:10.1016/S0040-4020(01)00454-9

TETRAHEDRON
Pergamon
Tetrahedron 57 &2001) 4841±4848
Synthesis of enantiomers of indanoxazolidinone based on the
lipase-catalyzed resolution of the corresponding
N-carbamylamino derivative
Masumi Suzuki, Chiaki Nagasawa and Takeshi Sugaip
Department of Chemistry, Keio University, Hiyoshi, Yokohama 223-8522, Japan
Received 26 March 2001; accepted 23 April 2001
AbstractÐEnantiomerically enriched &4R,5S)- and &4S,5R)-indano[1,2-d]oxazolidinones were enzymatically prepared from &^)-1-amino-
2-indanol. Racemic 1-&N0-chloroacetyl-N-carbamylamino)-2-indanol O-chloroacetate was hydrolyzed with immobilized Pseudomonas
cepacia lipase in the presence of b-cyclodextrin in acetone-buffer solution, to afford &1S,2R)-1-&N0-chloroacetyl-N-carbamylamino)-2-
indanol &90%e.e.) and the unreacted &1R,2S)-substrate &97%e.e.), in nearly quantitative yields. The deprotection provided enantiomers of
1-N-carbamylamino-2-indanol, the precursor of indanoxazolidinone, via nitrosation-deaminocyclization reaction. q 2001 Elsevier Science
Ltd. All rights reserved.
1. Introduction
Furthermore, the amino alcohol [&1S,2R)-2] has another
importance as the component of Crixivanw, a potent
protease inhibitor of HIV.2,6
Enantiomerically enriched forms of indanoxazolidinone
&1) [for example, &4S,5R)-form, Scheme 1] have been
developed as a unique and highly ef®cient chiral auxiliary
for the asymmetric aldol reaction.1 Many efforts have so far
been devoted toward the enantiomerically enriched forms of
cis-1-amino-2-indanol &2), the framework of 1.1a,b,2±4 Other
pioneering work in asymmetric syntheses has also been
carried out based on the open chain derivatives of 2 itself.5
Here we present an approach based on the enantiomeric
resolution of 1-N-carbamylamino-2-indanol &3), the direct
precursor of 1 through nitrosation-deaminocyclization
reaction,7 by way of lipase-catalyzed enantioselective
hydrolysis of the corresponding acyl precursor [&^)-4],
starting from racemic 1-amino-2-indanol [&^)-2].8
Scheme 1.
Keywords: enzymes and enzyme reactions; hydrolysis; nitroso compounds; oxazolidinones.
Corresponding author. Fax: 181-45-566-1697; e-mail: sugai@chem.keio.ac.jp
p
0040±4020/01/$ - see front matter q 2001 Elsevier Science Ltd. All rights reserved.
PII: S0040-4020&01)00454-9

4842
M. Suzuki et al. / Tetrahedron 57 -2001) 4841±4848
Scheme 2.
Table 1. Attempts for bisacetylation of N-carbamylamino indanol &3)
Entry
Reagent
2O, Py
2O, Py, DMAPc
2O, Et3N/DMFc
2O, AcONa
2O, AcOk
2O
Temp.
Time
Yield of 5 &%)
Major product
1
2
3
4
5
6
A
A
A
A
A
A
c
r.t.
r.t.
32 h
20 h
11 h
5 min
5 min
1.5 h
21
62
10
79
79
±
4a
5
4a
5
5
5
r.t.!508C
c
c
c
re¯ux
re¯ux
re¯ux
2. Results and discussion
acetylated by-product &5) was obtained in 21% yield. The
improved lipophilicity and lower melting point &see experi-
mental) of 5 compared to 4a, however, were attractive, so
that bisacetate would work as an alternative as the substrate
for lipase-catalyzed hydrolysis. This situation prompted us
to optimize the reaction conditions to obtain an enhanced
yield of 5.
Anumber of works have so far been reported in regard to
enzyme-catalyzed kinetic resolutions of related inter-
mediates. Most of them adopted the acylation of 2-indanols
bearing hydroxyl-,3a N-acetamido-3c and azido3g group on
the 1-position. Our plan as illustrated in Scheme 1 would
have advantages in the following points. The resolution of
direct precursors can exclude any loss of isomeric and
chemical purity of the resolved objectives. In this sense,
only one related example has been reported, however, the
yield was not satisfactory.3d In addition, the remarkable
advantage of enzyme-catalyzed hydrolysis, compared to
the reverse reaction such as acylation and/or transesteri®-
cation,9 is the essentially high speci®c activity of enzymes
in aqueous media as well as the irreversibility of the
reaction.
Table 1 summarizes the attempts toward ef®cient N0,O-bis-
acetylation. In the initial trial, the rate of reaction was still
very slow even when molar equivalent DMAP was added.
Moreover, further reacted side-products were given &entry
2). In this reaction, these unexpected by-products were
revealed to be N-decarbamylated derivatives &6a and 6b).
To suppress the formation of the undesired materials, the
reaction condition was modi®ed by modulating the nucleo-
philicity and basicity of reagents. Abrief treatment of 3 with
acetic anhydride and sodium or potassium acetate10 &entries
4 and 5) at a raised temperature was effective to give as high
as 79% yield of 5. Without a base &entry 6), the reaction was
slower than those observed in entries 4 and 5, and the ratio
of the desired product &5) did not improve, judging from its
TLC analysis.
Our initial candidate for the enzyme substrate was the carba-
mylamino acetate &4a). Toward this end, the carbamylamino
alcohol &3) was submitted to acetylation in a conventional
manner, as shown in Scheme 2. The yield of 4a was,
however, as low as 76%, and besides this, an N0,O-bis-
Scheme 3.

M. Suzuki et al. / Tetrahedron 57 -2001) 4841±4848
Table 2. The lipase-catalyzed hydrolysis of bisacetate &5)
4843
results are shown in Table 2. Although we con®rmed the
large differences between the enantiomers of acetate &5) on
both Pseudomonas cepacia lipase and Candida antarctica-
lipase, which had been successfully applied to the lipase-
catalyzed transesteri®cation on the structurally related
substrates,3 the rates of reaction were very low.
Entry
Lipase
Substrate
Time &day)
As expected, the introduction of an electron-withdrawing
group11 was very effective, as exempli®ed in such as chloro-
acetates 8 and 9, and the hydrolysis of the &1S,2R)-isomers
was highly accelerated &entries 2 and 3). Fortunately, still a
large difference in the rates of hydrolysis between the enan-
tiomers was observed &entries 2 and 4, 3 and 5). From these
results, in the practical sense, chloroacetates were expected
for lipase-catalyzed kinetic resolution of racemate. Further
investigation was continued with racemic N0,O-bischloro-
acetate [&1R*,2S*)-9] as the substrate.
1
2
3
4
5
1
2
3
4
Candida antarctica
&1S,2R)-5
&1R,2S)-5
^
2
^
2
^
2
^
2
1
2
^
2
1
2
1
2
1
2
1
2
Pseudomonas cepacia &1S,2R)-5
&1R,2S)-5
Table 3. The effect of the electron-withdrawing group on the lipase-
catalyzed hydrolysis
As a considerable lowering of pH was observed even in an
acetone-buffer in the preliminary experiments &Table 3),
due to the liberation of a strong acid &chloroacetate), the
racemic substrate &9) was incubated under pH-controlling
condition &at 7.0±8.0). The reaction, however, was sluggish
and even after 3 days, the conversion was as low as 25%.
The reason for such low reactivity was ascribable to the very
low solubility of the racemate of 9. The melting point of the
racemate &172.9±173.48C) was much higher than that of
the pure enantiomer &123.4±123.98C). An X-ray crystallo-
graphic analysis revealed a large difference in the hydrogen-
bonding network between the racemate and the enantio-
merically pure form, which led to a denser packing of mole-
cules in the racemic crystal.12
Entry
Substrate
Product
Conversion &%)
Final pH
1
2
3
4
5
&1S,2R)-5
&1S,2S)-8
&1S,2R)-9
&1R,2S)-8
&1R,2S)-9
7
7
10
7
2.7
52.9
67.1
1.8
6.3
3.4
3.1
5.6
5.7
10
2.1
Under the same protocol, chloroacetates &8) and &9), which
would have enhanced reactivity toward lipase-catalyzed
hydrolysis,11 were prepared &Scheme 3). Selective de-O-
acetylation of &5) provided N0-acetylcarbamylamino alcohol
&7), which was then chloroacetylated to give N0-acetyl-O-
chloroacetylated substrate &8). On the other hand, a direct
treatment of carbamylamino alcohol &3) with chloroacetic
anhydride gave N0,O-bischloroacetate &9) in 95% yield.
Under either condition for the chloroacetylation as above,
no decarbamylation was observed even under as high as
858C.
The hydrolysis of &^)-8 was faster than that of &^)-9, as
expected from its lower melting point &164.2±164.88C)
compared to 9 &see the previous paragraph). Due to the
accessibility of bischloroacetate [&^)-9] by the simple
preparation from &^)-3, however, the substrate was ®xed
to be &^)-9, and the reaction conditions were extensively
elaborated as shown in Table 4 to overcome the problem
caused by this low solubility.
First, the ratio of a co-solvent &acetone) to water was
increased, expecting enhanced dissolution of the substrate
&entries 2 and 3). The medium range of concentration of
acetone &42±67%) only resulted in a loss of activity of
lipase. At a higher ratio &83%), however, the reaction
With the substrates in hand, the rates of lipase-catalyzed
hydrolysis between the enantiomers were compared. The
Table 4. The effect of the organic co-solvent on the lipase-catalyzed hydrolysis
Entry
Sub. conc. &w/v%)
Acetone/buffer
State
Reaction time &h)
Conversion &%)
1
2
3
4
5
4.2
2.0
2.7
3.3
4.2
1:14
1:1.0
1:0.5
1:0.2
1:1.4
Susp.
Susp.
Susp.
Sol.
71
66
68
72
37
25.1
25.9
11.6
47.9
3.2
Susp.a
a
DMF precipitation.13

4844
M. Suzuki et al. / Tetrahedron 57 -2001) 4841±4848
Table 5. The effect of additives on the lipase-catalyzed hydrolysis
Entry
Sub. conc. &w/v%)
Cosolvent
Cosolvent-water
Additive
None
None
Triton X-100
b-cyclodextrin
Effect
1
2
3
4
2.7
10.0
3.0
Acetone
DMF
Acetone
Acetone
1:0.13
1:0.14
1:0.13
1:0.13
No reaction
2
1
2.7
smoothly proceeded, as expected from the homogeneity of
the reaction mixture &entry 4). In the present example, the
state of substrates, the completely dissolved form, was
important and the ef®ciency was not affected by the surface
of the solid substrate. Indeed, pre-treatment of the substrate
with DMF &DMF aggregate amorphous formation)13 had no
effect in our present case &entry 5).
The unreacted starting material &1R,2S)-9, and the corre-
sponding alcohol &1S,2R)-10 were obtained in 47 and 51%
yield, respectively. Both products were ef®ciently converted
to the enantiomers of 3, the precursor for cyclization, by
treatment with a sodium methoxide-methanol solution.
After the nitrous-acid catalyzed cyclization reaction,7 the
e.e.s were determined at the stage of the oxazolidinones
&1) by the GLC analysis.19 The e.e.s of &1R,2S)-9 &97%)
and &1S,2R)-10 &90%) indicated that the conversion and E
value20 of the Pseudomonas lipase-catalyzed reaction were
51.8 and 79, respectively &Scheme 4).
Further attempts for the enhancement of the rate of
hydrolysis were continued, as the retention of high enzyme
activity in organic solvent was indexed as shown in Table 5.
From this standpoint, the immobilization of Pseudomonas
lipase on Toyonite 200-M14 was very effective &entry 1).
Changing the co-solvent from acetone to DMF, which
would lower the limit of the organic solvent to make a
homogeneous state, however, caused the complete
denaturation of enzyme &entry 2).15 At this stage, as a
different approach, the effect of additives was also
examined, expecting an ef®cient transfer of the substrate
which stayed in the organic solvent to the hydrophilic
surroundings of the enzyme.16 In contrast to a detergent
such as Triton X-100 &entry 3),17 the addition of b-cyclo-
dextrin &0.1±1.0 equiv.),18 had a remarkable accelerating
effect &entry 4).
As we mentioned earlier, pure &4S,5R)-enantiomer of
indanoxazolidinone &1) has more importance as the building
block of Crixivanw. The enantiomerically enriched alcohol,
&1S,2R)-10 &for example, 81%e.e.) was allowed to give
the desired product by reacylation and the subsequent
hydrolysis with P. cepacia lipase as shown in Scheme 5.
In this case, due to the low melting point of the substrate
&115.5±116.48C), the hydrolysis proceeded more smoothly,
even under the low concentration of an organic co-solvent
and with native &non-immobilized) lipase.
In summary, both enantiomers of indanoxazolidinone were
synthesized from racemic 1-amino-2-indanol via immo-
bilized lipase PS-catalyzed kinetic resolution. The highly
crystalline nature of the substrate of the enzymatic
Based on the above-mentioned elaborated condition,
upscaling of the reaction &substrate: 2 g) was attempted.
Scheme 4.

M. Suzuki et al. / Tetrahedron 57 -2001) 4841±4848
4845
Scheme 5.
hydrolysis brought about the very low reactivity under
conventional conditions, and elaboration of the reaction
conditions, especially in regard to the dissolution of the
substrate, overcame this problem.
5.20 mmol]. After stirring for 5 min at re¯ux, the reaction
mixture was poured into ice, and the mixture was stirred for
30 min to destroy excess acetic anhydride. The mixture was
concentrated in vacuo, then toluene was added and further
evaporated until the trace of AcOH was removed. The resi-
due was puri®ed by silica gel column chromatography
&150 g). Elution with hexane-AcOEt &2:3 to 3:7) afforded
5 &79% yield). Recrystallization from Et2O provided color-
less prisms. Mp 128.0±128.48C; 1H NMR &400 MHz,
CDCl3) d 10.19 &s, 1H), 9.03 &d, 1H, Jˆ8.3 Hz), 7.30 &m,
4H), 5.68±5.61 &m, 2H), 3.25 &dd, 1H, Jˆ4.9, 17.1 Hz), 3.06
&d, 1H, Jˆ17.1 Hz), 2.15 &s, 3H), 2.06 &s, 3H); IR &KBr)
3300, 3239, 3115, 2981, 2944, 1738, 1715, 1691, 1550,
1510, 1375, 1259, 1243, 1040 cm21. Anal. Calcd. for
C14H16N2O4: C, 60.86; H, 5.84; N, 10.14. Found: C,
60.87; H, 5.86; N, 10.16.
3. Experimental
3.1. General
All mps were uncorrected. IR spectra were measured as
®lms for oils and KBr disks for solids, on a JASCO FT/
IR-410 spectrometer. H NMR spectra were measured at
1
270 MHz on a JEOL JNM EX-270 or at 400 MHz on a
JEOL JNM GX-400 spectrometer. 13C-NMR spectra were
measured at 100 MHz on a JEOL JNM GX-400 spectro-
meter. Mass spectra were recorded on Hitachi M-80B spec-
trometer at 70 eV. GLC data were recorded on a GC-353
&GL Sciences Inc.) gas chromatograph. Optical rotations
were recorded on a JASCO DIP 360 polarimeter. Wako
Gel B-5F and silica gel 60 K070-WH &70-230 mesh) of
Katayama Chemical Co. were used for preparative TLC
and column chromatography, respectively.
In the course of the preparation of 5, 1-N-carbamylamino-2-
indanol O-acetate &4a), 1-acetamide-2-indanol O-acetate
&6a) and 1-N,N-diacetylamino-2-indanol O-acetate &6b)
were obtained as by-products. 4a &Fine needles from
1
AcOEt-EtOH): mp 190.2±191.08C; H NMR &400 MHz,
CD3OD) d 7.23 &m, 4H), 5.55 &m, 1H), 5.36 &d, 1H, Jˆ
4.9 Hz), 3.18 &dd, 1H, Jˆ5.4, 17.1 Hz), 2.95 &dd, 1H, Jˆ
1.5, 17.1 Hz), 2.00 &s, 3H); IR &KBr) 3491, 3335, 3291,
3.1.1. *1R*,2S*)-*^)-1-N-Carbamylamino-2-indanol 3.
To a solution of &1R*,2S*)-&^)-1-amimo-2-indanol &2,
1.00 g, 6.70 mmol) in 2 M HCl &4.36 mL, 8.71 mmol) was
added potassium cyanate &0.82 g, 10.05 mmol). The mixture
was stirred at room temperature for 5 min, a white solid was
deposited. After stirring for another 30 min, the precipitate
was ®ltered and washed with ice-cooled water. The
carbamylamino alcohol &3) was obtained in 95% yield.
Mp 235.2±235.58C; 1H NMR &400 MHz, DMSO-d6) d
7.12 &m, 4H), 6.14 &d, 1H, Jˆ8.8 Hz), 5.74 &s, 2H), 5.09
&d, 1H, Jˆ3.9 Hz), 4.98 &m, 1H), 4.35 &dd, 1H, Jˆ3.9,
8.8 Hz), 2.99 &dd, 1H, Jˆ4.9, 16.1 Hz), 2.75 &d, 1H, Jˆ
16.1 Hz); 13C-NMR &100 MHz, DMSO-d6) d 39.8, 57.4,
72.3, 123.7, 124.7, 126.0, 126.8, 140.2, 143.5, 158.6; IR
&KBr) 3441, 3328, 1649, 1629, 1457, 1381, 1297, 1177,
1052 cm21. Anal. Calcd. for C10H12N2O2: C, 62.49; H,
6.29; N, 14.57. Found: C, 62.40; H, 6.33; N, 14.59. The
enantiomers [&1S,2R)- and &1R,2S)-form] were prepared in
the same manner.
1734, 1720, 1653, 1597, 1566, 1376, 1245, 1041 cm21
.
Anal. Calcd. for C12H14N2O3: C, 61.53; H, 6.02; N, 11.96.
Found: C, 61.18; H, 6.04; N, 11.62. 6a &Colorless solid):
1H-NMR &400 MHz, CDCl3) d 7.25 &m, 4H), 5.98 &d, 1H,
Jˆ9.3 Hz), 5.68 &dd, 1H, Jˆ5.4, 9.3 Hz), 5.55 &ddd, 1H,
Jˆ1.5, 5.4, 5.4 Hz), 3.21 &dd, 1H, Jˆ5.4, 17.1 Hz), 3.01
&dd, 1H, Jˆ1.5, 17.1 Hz), 2.10 &s, 3H), 2.02 &s, 3H); IR
&KBr) 3303, 3072, 1732, 1651, 1550, 1377, 1288, 1244,
1
1211, 1043 cm21. H NMR spectrum was identical with
an authentic sample prepared from &1S,2R)-aminoindanol
&2). 6b &Colorless oil): 1H NMR &400 MHz, CDCl3) d
7.19 &m, 3H), 7.09 &d, 1H, Jˆ7.3 Hz), 5.92 &d, 1H,
Jˆ8.3 Hz), 5.53 &m, 1H), 3.31 &dd, 1H, Jˆ7.8, 16.8 Hz),
3.25 &dd, 1H, Jˆ5.4, 16.8 Hz), 2.22 &br s, 6H), 1.96 &s,
3H); 13C-NMR &100 MHz, CDCl3) d 20.8, 27.0, 38.3,
60.7, 72.7, 123.8, 125.7, 127.8, 129.1, 139.2, 140.5, 171.3,
175.0; IR &NaCl) 3023, 2937, 1738, 1704, 1432, 1368, 1239,
1070 cm21. Anal. Calcd. for C15H17NO4: C, 65.44; H, 6.22;
N, 5.09. Found: C, 65.44; H, 6.24; N, 5.16.
3.1.2. *1R,2S)-1-*N'-Acetyl-N-carbamylamino)-2-indanol
O-acetate 5. To a re¯uxing suspension of potassium acetate
&0.37 g, 3.80 mmol) in acetic anhydride &15.0 mL) was
added N-carbamylamino alcohol [&1R,2S)-3, 1.00 g,
In the same manner as described above, &1S,2R)-5 was
obtained from &1S,2R)-3. H NMR spectrum was identical
with that of &1R,2S)-5.
1

4846
M. Suzuki et al. / Tetrahedron 57 -2001) 4841±4848
3.1.3. *1S,2R)-1-*N'-Acetyl-N-carbamylamino)-2-indanol 7.
To a solution of &1S,2R)-5 &400 mg, 1.45 mmol) in MeOH
&4.0 mL) was added NaOMe solution &0.11 M in MeOH,
1.33 mL, 0.15 mmol) dropwise at 08C. After stirring for
1 h at room temperature, the reaction was quenched with
aqueous NH4Cl and extracted with AcOEt three times. The
combined organic layer was dried over Na2SO4 and concen-
trated in vacuo. The residue was puri®ed by silica gel
column chromatography &14 g). Elution with AcOEt
afforded &1S,2R)-7 &74% yield). Recrystallization from
hexane-EtOH provided colorless needles. Mp 181.5±
152.6, 166.8, 167.6; IR &KBr) 3300, 3235, 3112, 2973,
1753, 1724, 1694, 1561, 1513, 1404, 1316, 1250, 1200,
1176, 1042 cm21. Anal. Calcd. for C14H14Cl2N2O4: C,
48.71; H, 4.09; N, 8.12. Found: C, 48.75; H, 4.02; N, 8.07.
In the same manner, &1S,2R)-9 was prepared. Recrystalli-
zation from hexane-AcOEt provided colorless needles; mp.
20
123.4±123.98C; [a]D ˆ237.2 &c 1.01, CHCl3); IR &KBr)
3291, 3245, 3141, 2953, 1751, 1728, 1702, 1550, 1499,
1405, 1316, 1248, 1199, 1173, 1057 cm21 1H NMR
.
spectrum was same as &^)-9.
1
181.98C; H NMR &270 MHz, CDCl3) d 9.76 &s, 1H), 8.94
&d, 1H, Jˆ7.4 Hz), 7.24 &m, 4H), 5.24 &dd, 1H, Jˆ5.2,
7.4 Hz), 4.59 &br s, 1H), 3.11 &dd, 1H, Jˆ5.2, 16.5 Hz),
2.92 &dd, 1H, Jˆ1.5, 16.5 Hz), 2.59 &br s, 1H), 2.03 &s,
3H); IR &KBr) 3518, 3406, 3230, 3241, 3133, 2964, 1690,
1676, 1571, 1536, 1255, 1044 cm21. Anal. Calcd. for
C12H14N2O3: C, 61.53; H, 6.02; N, 11.96. Found: C,
61.45; H, 6.01; N, 11.96.
The enantiomer [&1R,2S)-form] was also prepared in the
same manner. H NMR spectrum was identical with that
of &^)-9.
1
3.2. Chirazymew L-2 and lipase PS catalyzed hydrolysis
of *1R*,2S*)-5
The rates of hydrolyses of both enantiomers of 5 with C.
antarcticalipase &Chirazymew L-2) and P. cepacia lipase
&Amano PS) were compared as follows.
3.1.4. *1S,2R)-1-*N'-Acetyl-N-carbamylamino)-2-indanol
O-chloroacetate 8. The mixture of &1S,2R)-7 *81 mg,
0.35 mmol), chloroacetic anhydride &117 mg, 0.69 mmol),
pyridine &1.4 mL) and MS 3A&ca. 400 mg) was stirred for
1 h at room temperature. After quenching with pH 7.0 phos-
phate buffer, the reaction mixture was ®ltered to remove
insoluble materials, and the ®ltrate was extracted with
AcOEt three times. The combined organic layer was
concentrated in vacuo. To the residue was added toluene
and evaporated in vacuo until pyridine was removed. The
residue was puri®ed by silica gel column chromatography
&13 g). Elution with hexane-AcOEt &2:3) afforded &1S,2R)-8
&quant.). Recrystallization from hexane-AcOEt provided
To a solution of N0,O-diacetyl-N-carbamylamino alcohol
[&1S,2R)- or &1R,2S)-5, each 50 mg] in acetone &0.4 mL)
and pH 7.0 phosphate buffer &0.1 M, 0.6 mL) was added
lipase &10 mg). The reaction mixture was stirred at room
temperature. The conversion was roughly estimated on the
basis of TLC analysis &AcOEt, Rfˆ0.56 for the substrate 5,
0.30 for the product 7).
3.3. Comparison the rate of lipase PS catalyzed
hydrolysis between *1S,2R)-5, *1R*,2S*)-8 and
*1R*,2S*)-9
1
colorless plates. Mp 119.6±120.18C; H NMR &270 MHz,
CDCl3) d 10.11 &s, 1H), 8.97 &d, 1H, Jˆ8.9 Hz), 7.31 &m,
4H), 5.77 &dd, 1H, Jˆ5.1, 5.2 Hz), 5.66 &dd, 1H, Jˆ5.2,
8.9 Hz), 4.08 &dd, 2H, Jˆ15.3 Hz), 3.30 &dd, 1H, Jˆ5.1,
17.2 Hz), 3.11 &d, 1H, Jˆ17.2 Hz), 2.16 &s, 3H); IR &KBr)
3231, 3118, 2960, 1765, 1691, 1550, 1481, 1239, 1173,
1037 cm21. Anal. Calcd. for C14H15ClN2O4: C, 54.11; H,
4.87; N, 9.02. Found: C, 54.01; H, 4.68; N, 9.00.
The substrate &50 mg), acetone &0.5 mL), pH 7.0 phosphate
buffer &0.1 M, 0.7 mL) and lipase PS &10 mg) were mixed
and stirred for 24 h at room temperature. To the reaction
mixture was added brine and extracted with AcOEt 3±5
times. The combined organic layer was washed with
brine, dried over Na2SO4 and concentrated in vacuo. The
1
residue was dissolved in CDCl3 and its H NMR spectrum
In the same manner as described above, &1R,2S)-8 was
obtained. Its H NMR spectrum was identical with that of
&1S,2R)-8.
was measured. The ratios between expecting products 7, 10
and the starting materials respectively, were estimated by
comparing the signals of the substrates [H-1 and H-2 &d
5.68±5.61 for 5, 5.77 and 5.66 for 8, 5.70 and 5.60 for 9)]
with those of the products &d 5.24 and 4.59 for 7, 5.29 and
4.63 for 10).
1
3.1.5.
*1R*,2S*)-*^)-1-*N'-Chloroacetyl-N-carbamyl-
amino)-2-indanol O-chloroacetate 9. Amixture of
N-carbamylamino alcohol &3, 4.50 g, 23.4 mmol) and
chloroacetic anhydride &17.0 g, 99.5 mmol) was heated at
858C and stirred for 10 min. The reaction mixture was
poured into pH 7.0 phosphate buffer and stirred for 1 h,
white solids were gradually deposited. The precipitate was
®ltered and washed with aqueous NaHCO3 and water to give
9 &95% yield). This was employed for the next step without
further puri®cation. Analytical sample &^)-9 &colorless
needles from AcOEt): mp 172.9±173.48C; 1H NMR
&400 MHz, CDCl3) d 9.17 &br s, 1H), 8.56 &d, 1H, Jˆ
8.8 Hz), 7.23 &m, 4H), 5.70 &m, 1H), 5.60 &dd, 1H, Jˆ5.4,
8.8 Hz), 4.07 &s, 2H), 4.03 &d, 1H, Jˆ15.1 Hz), 3.97 &d, 1H,
Jˆ15.1 Hz), 3.23 &dd, 1H, Jˆ4.9, 17.1 Hz), 3.04 &d, 1H,
Jˆ17.1 Hz); 13C-NMR &100 MHz, CDCl3) d 37.2, 41.0,
42.5, 56.5, 77.1, 123.9, 125.0, 127.5, 128.6, 138.8, 139.3,
3.4. Immobilization of lipase PS on toyonite 200-M
Asuspension of commercial lipase PS &12.50 g) in pH 7.0
phosphate buffer &0.1 M, 50.0 mL) was stirred for 30 min.
The mixture was centrifuged, and the supernatant was
passed through an Amicon YM10 ultra®ltration membrane
&62 mmf) until the volume of the solution inside the cell
reached 7.6 mL. To this concentrated protein solution was
added Toyonite 200-M &268 mg) and the mixture was
shaken for 14 h at 258C under argon. It was ®ltered and
the residue was allowed to dry in vacuo to afford immobi-
lized lipase PS &300 mg). The weight of protein retained on
the support was estimated to be 23 mg by the CBB method,
being calibrated with g-globulin as the standard.

M. Suzuki et al. / Tetrahedron 57 -2001) 4841±4848
4847
3.5. Effect of additives in immobilized lipase PS
catalyzed hydrolysis
acetyl-N-carbamylamino alcohol 9 was converted to
&4R,5S)-1 &2 steps, quant.). On the basis of GLC analysis,
the e.e. was estimated to be 96.7%.
To a mixture of &^)-9 &400 mg, 1.16 mmol) and an additive
&Triton X-100 or b-CD) in acetone-water, was added immo-
bilized lipase PS &17 mg). While keeping its pH above 6.6
by a pH controller, the reaction mixture was stirred at room
temperature until ca. 50% conversion. The conversion was
checked by 1H NMR analysis &CDCl3) of a small portion of
the reaction mixture, in the same manner as described above.
3.7. Enhancement of the e.e. of *1S,2R)-10 by repetition
of lipase-catalyzed hydrolysis
An enantiomerically enriched form of &1S,2R)-10
&80.9%e.e.), which obtained by the lipase-catalyzed hydro-
lysis as mentioned above, was chloroacetylated again to
give &1S,2R)-9. This &50 mg, 0.14 mmol) was suspended
in acetone &0.5 mL) and pH 7.0 phosphate buffer &0.1 M,
0.7 mL), and was added native &non-immobilized) lipase
PS &10 mg). While controlling its pH about 6 by addition
of aqueous KOH, the reaction mixture was stirred for 20 h at
room temperature. Conventional workup and chromato-
graphic separation [hexane-AcOEt &2:3)] afforded &1S,2R)-
9 &5.5%e.e., 20% yield) and &1S,2R)-10 &99.9%e.e., 80%
yield). The e.e.s were determined at the stage of oxazo-
lidinones, in the same manner as described above.
3.6. Upscaling of the hydrolysis of *^)-9 with
immobilized lipase PS
To a solution of &^)-9 &2.00 g, 5.79 mmol) and b-CD
&0.66 g, 0.58 mmol) in acetone &65.0 mL) and pH 7.0 phos-
phate buffer &0.01M, 6.6 mL), was added immobilized
lipase PS &200 mg). While keeping its pH above 6.9 by a
pH controller, the reaction mixture was stirred for 15 h at
358C. The immobilized lipase was ®ltered off and the ®ltrate
was evaporated in vacuo to remove acetone. The mixture
was extracted with AcOEt ®ve times. The combined organic
layer was dried over Na2SO4 and concentrated in vacuo. The
residue was puri®ed by silica gel column chromatography
&300 g). Elution with CHCl3-AcOEt &1:0 to 1:9) afforded
&1R,2S)-9 &47% yield) and &1S,2R)-10 &51% yield). Both
products were converted to indanoxazolidinones &1) without
further puri®cation and the e.e.s were determined. &1S,2R)-
10: 1H NMR &400 MHz, CDCl3) d 8.68 &s, 1H), 8.62 &d, 1H,
Jˆ7.2 Hz), 7.22 &m, 4H), 5.29 &dd, 1H, Jˆ5.4, 7.2 Hz), 4.63
&dddd, 1H, Jˆ2.4, 5.4, 5.4, 5.4 Hz), 3.14 &dd, 1H, Jˆ5.4,
16.6 Hz), 2.93 &dd, 1H, Jˆ2.4, 16.6 Hz), 1.98 &d, 1H, Jˆ
5.4 Hz); MS m/zˆ269 &M1). Recrystallization from AcOEt
gave an analytical sample as colorless needles. Anal. Calcd.
for C12H13ClN2O3: C, 53.64; H, 4.88; N, 10.43. Found: C,
53.79; H, 4.89; N, 10.31.
Acknowledgements
The authors thank Amano Enzyme Co., Roche Diagnostics
Co. and Toyo Denka Kogyo, for the generous gifts of lipase
PS, Chirazymew L-2 and Toyonite 200-M, respectively. We
are grateful to Professor Shigeru Ohba of Keio University
for X-ray analysis. We also express our thanks to Dr Harumi
Kaga of Hokkaido National Industrial Research Institute
and Professor Tadashi Kometani of Toyama National
College of Technology for their discussion. This work
was accomplished as `Science and Technology Program
on Molecules, Supra-Molecules and Supra-Structured
Materials' of an Academic Frontier Promotional Project,
by Japan's Ministry of Education, Culture, Sports, Science
and Technology, and acknowledged with thanks. This work
was also supported by a Grant-in-Aid for Scienti®c
Research &No. 12660120).
3.6.1. *4S,5R)-Indano[1,2-d]oxazolidinone 1. To a solution
of &1S,2R)-10 &75 mg, 0.28 mmol) in MeOH &3.0 mL) was
added NaOMe solution &0.11 M, 0.25 mL, 0.03 mmol) at
room temperature. The reaction mixture was stirred over-
night and quenched with H2O. After evaporating the organic
solvent in vacuo, 2 M HCl &1.5 mL, 3.0 mmol) and AcOH
&7.0 mL) were added to the residue. To the solution was
added NaNO2 &31 mg, 0.45 mmol) and the mixture
was stirred for 1 h at room temperature. To the reaction
was added aqueous KOH until the pH of the mixture was
reached 10 and the mixture was extracted with CHCl3 three
times. The combined organic layer was washed with
aqueous NaHCO3 and brine, dried over Na2SO4, concen-
trated in vacuo. &4S,5R)-Oxazolidinone &1) was obtained
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