Biocatalytic syntheses of protected D-mannose-d5, D-mannose-d7, D-mannitol-2,3,4,5,6-d5, and D-mannitol-1,1,2,3,4,5,6,6-d8

Full text of DOI:10.1021/jo951666s

J . Org. Chem. 1996, 61, 4151-4153
4151
diols.10 Among these accomplishments are the brief
preparations of cyclitols,11 aminocyclitols,12 monosaccha-
rides,13 aza sugars,14 a novel alkaloid kifunensine,15
pseudo-sugars,16 and, most recently, amino sugars.17 For
this study deuterated derivatives of the four biologically
important hexoses, D-glucose, D-mannose, D-galactose,
and D-fucose, were selected as targets in which the
synthesis of perdeuteriomannose would be evaluated to
determine the feasibility of this approach. Herein we
report the first application of our general protocol to the
synthesis of d5- and d7-mannoses and perdeuterated
mannitol derivatives via toluene dioxygenase-mediated
oxidation of d5-halobenzenes.
Bioca ta lytic Syn th eses of P r otected
D-Ma n n ose-d 5, D-Ma n n ose-d 7,
D-Ma n n itol-2,3,4,5,6-d 5, a n d
D-Ma n n itol-1,1,2,3,4,5,6,6-d 8
Tomas Hudlicky,* Kevin K. Pitzer,
Michele R. Stabile, and Andrew J . Thorpe
Department of Chemistry, University of Florida,
Gainesville, Florida 32611-7200
Gregory M. Whited
Genencor International, Inc., 180 Kimball Way,
South San Francisco, California 94080
Perdeuteriochlorobenzene (1a ) was subjected to whole
cell oxidation by Pseudomonas putida 39/D18 or Escheri-
chia coli J M109(pDTG601)19 as previously described,20
yielding the corresponding halocyclohexadiene-cis-dihy-
drodiol 2a in approximate yields of 1 g/L (Scheme 1).
Protection of diols 2a and 2b21 as their acetonides
followed by osmylation provided the key intermediates:
cis-diol 4a as a colorless oil (85% overall yield) and cis-
diol 4b as a white solid (68% overall yield). Diols 4 were
subjected to ozonolysis and reductive workup, giving a
mixture of compounds which were acetylated under
standard conditions providing a mixture of anomers of
acetyl 4,6-diacetyl-2,3-O-isopropylidene-D-mannoside-
2,3,4,5,6-d5 (5) as the major identifiable product. This
compound was isolated as a 3:2 mixture of C6 epimers,
indicated by a heteronuclear multiple quantum correla-
tion experiment (HMQC). Alternatively, ozonolysis of
diol 4a and subsequent reduction with sodium borodeu-
teride gave the intermediate alcohol which was acetyl-
ated to yield acetyl 4,6-diacetyl-2,3-O-isopropylidene-D-
mannoside-1,2,3,4,5,6,6-d7 (6). In order to provide a
viable route to perdeuteriomannose, diol 4a was sub-
jected to a similar ozonylitic/reductive sequence employ-
ing sodium borodeuteride to give the intermediate triol
which was converted to the glycoside 7 in two steps.
Received September 11, 1995 (Revised Manuscript Received
March 14, 1996 )
Specifically labeled carbohydrates constitute a group
of compounds that are both difficult to prepare and highly
desirable for various biological studies. For example,
labeled glucoses were utilized to help obtain dynamic
measurements of the cerebral pentose phosphate path-
way both in vivo1 and in cultured cells.2 Deuterated
sugars have been applied to study the pathway of hepatic
glycogen synthesis in rats3 and in humans.4 d7-Glucose
was employed to study the biodistribution of glucose in
rats,5 and a labeled galactitol was used to help diagnose
disorders of galactose metabolism of fetuses.6 Finally,
labeled glucose has been used to study glucose metabo-
lism in animals and man.7
Deuterated carbohydrates that are available com-
mercially include, but are not limited to, such sugars as
D-glucose-6,6-d2 ($86.60/500 mg), D-glucose-1-d ($68.49/
250 mg), and D-glucose-2-d ($169.40/250 mg). Perfluori-
nated derivatives are not available presumably because
of the expected facile elimination of HF from unprotected
species. Of the eight simple D-sugars, only glucose had
been previously synthesized as its hepta-C-deuteride.8
Analogs of the simple D-sugars will be eminently useful
in protein binding studies as well as in instances where
deuterium incorporation originating in carbohydrate
metabolism is desired. Deuterated sugars will also be
helpful in understanding the dynamics of carbohydrates
in solution.9
1
Comparison of the H NMR of this molecule 7 with that
(10) For recent reviews concerning halocyclohexadiene-cis-diols,
see: (a) Hudlicky, T.; Reed, J . W. Adv. Asymm. Synth. 1995, 1, 271.
(b) Brown, S. M.; Hudlicky, T. In Organic Synthesis: Theory and
Practice; Hudlicky, T., Ed; J AI Press: Greenwich, CT, 1992; Vol. 2, p
113. (c) Carless, H. A. J . Tetrahedron: Asymmetry 1992, 795. (d)
Widdowson, D. A.; Ribbons, D. A.; Thomas, S. D. J anssen Chim. Acta
1990, 8, 3.
(11) (a) Mandel, M.; Hudlicky, T. J . Org. Chem. 1993, 58, 2331. (b)
Mandel, M.; Hudlicky, T. J . Chem. Soc., Perkin Trans. 1 1993, 741. (c)
Hudlicky, T.; Luna, H.; Olivo, H. F.; Anderson, C.; Nugent, T.; Price,
J . D. J . Chem. Soc., Perkin Trans. 1 1991, 2907. (d) Hudlicky, T.; Price,
J . D.; Luna, H.; Anderson, C. M. Synlett 1990, 309. (e) Hudlicky, T.;
Price, J . D.; Rulin, F.; Tsunoda, T. J . Am. Chem. Soc. 1990, 112, 9439.
(12) Hudlicky, T.; Olivo, H. F.; McKibben, B. J . Am. Chem. Soc.
1994, 116, 5108.
(13) (a) Hudlicky, T.; Mandel, M.; Rouden, J .; Lee, R. S.; Bachmann,
B.; Dudding, T.; Yost, K. J .; Merola, J . J . Chem. Soc., Perkin Trans. 1
1994, 1553. (b) Hudlicky, T.; Price, J . D.; Rulin, F. J . Org. Chem. 1990
55, 4683. (c) Hudlicky, T.; Luna, H.; Price, J . D.; Rulin, F. Tetrahedron
Lett. 1989, 30, 4053.
(14) Hudlicky, T.; Rouden, J .; Luna, H. J . Org. Chem. 1993, 58, 985.
(15) (a) Hudlicky, T.; Rouden, J .; Luna, H.; Allen, S. J . Am. Chem.
Soc. 1994, 116, 5099. (b) Hudlicky, T.; Rouden, J .; Luna, H. J . Org.
Chem. 1993, 58, 985.
For the past several years we have devoted consider-
able time and effort to the development of concise
syntheses of carbohydrates from halocyclohexadiene-cis-
(1) Ben-Yoseph, O.; Camp, D. M.; Robinson, T. E.; Ross, B. D. J .
Neurochem. 1995, 64, 1336.
(2) Ross, B. D.; Kingsley, P. B.; Ben-Yoseph, O. Biochem. J . 1994,
302, 31.
(3) Goodman, M. N.; Masuoka, L. K.; DeRopp, J . S.; J ones, A. D.
Biochem. Biophys. Res. Commun. 1989, 159, 522.
(4) Argoud, G. M.; Schade, D. S.; Eaton, R. P. Am. J . Physiol. 1987,
252, E606.
(5) Gatley, S. J .; Wess, M. M.; Govini, P. L.; Wagner, A.; Katz, J . J .;
Friedman, A. M. J . Nucl. Med. 1986, 27, 388.
(6) J akobs, C.; Warner, T. G.; Sweetman, L.; Nyhan, W. L. Ped. Res.
1984, 18, 714.
(16) Entwistle, D. A.; Hudlicky, T. Tetrahedron Lett. 1995, 36, 2591.
(17) Pitzer, K.; Hudlicky, T. Synlett 1995, 803.
(18) (a) Gibson, D. T.; Koch, J . R.; Kallio, R. E. Biochemistry 1968,
7, 2653. (b) Gibson, D.; Hemsley, M.; Yoshioka, H.; Mabry, T.
Biochemistry 1970, 9, 1626.
(19) Zylstra, G. J .; Gibson, D. T. J . Biol. Chem. 1989, 264, 14940.
(20) Hudlicky, T.; Boros, C. H.; Boros, E. E. Synthesis 1992, 174.
(21) Diol 2b is derived from perdeuteriobromobenzene which was
obtained from Genencor International.
(7) Bier, D. M.; Arnold, K. J .; Sherman, W. R.; Holland, W. H.;
Holmes, W. F.; Kipnis, D. M. Diabetes 1977, 26, 1005.
(8) (a) Koch, H. J .; Stuart, R. S. Carbohydr. Res. 1978, 67, 341. (b)
Koch, H. J .; Stuart, R. S. Carbohydr. Res. 1978, 64, 127. (c) Koch, H.
J .; Stuart, R. S. Carbohydr. Res. 1978, 59, C1. (d) This sugar is
available commercially.
(9) Gervay, J ., University of Arizona, private communication.
S0022-3263(95)01666-5 CCC: $12.00 © 1996 American Chemical Society

4152 J . Org. Chem., Vol. 61, No. 12, 1996
Notes
Sch em e 1a
2,3,4,5,6-d5 (11) (44% overall yield from 8) and ((tert-
butyldimethylsilyl)oxy)-2,3-O-isopropylidene-D-mannitol-
1,1,2,3,4,5,6,6-d8 (12) (34% overall yield from 8).23
In conclusion, the synthesis of perdeuterated carbohy-
drates from d5-halobenzenes proved to be reasonably
facile, certainly when compared to any preparations of
the title compounds from natural sugars by oxidation/
reduction sequences. Studies leading to other labeled
carbohydrates by application of the previously disclosed
general method of synthesis coupled with biooxidation
of specificaly labeled substrates are in progress.
Exp er im en ta l Section
General experimental procedures have been published pre-
viously.11c
(1S ,2S )-3-C h lo r o c y c lo h e x a -3,5-d ie n e -1,2-d io l-1,2,4,
5,6-d 5 (2a ). Perdeuterated diol 2a was used without purifica-
tion:mp 91-94 °C; [R]26 +59.0 (c 1.06, CHCl3); IR (KBr) cm-1
D
3294, 2777, 2191, 1604, 1566; 1H NMR (CDCl3) δ 4.4 (s, OH),
4.4 (s, OH); 13C NMR (CDCl3) δ 135.9, 131.1, 130.8, 130.4, 123.5,
123.1, 122.8, 122.5, 122.1, 121.8, 71.8, 71.5, 71.2, 70.3, 70.0, 69.7;
MS (EI) m/z (rel intensity) 151 (M+) (60), 133 (23), 105 (100), 85
(21), 70 (40); HRMS calcd for C6D5H2ClO2 151.0448, found
151.0450, error 1.3 ppm.
a
Reagents: (i) Pseudomonas putida 39/D or Escherichia coli
J M109; (ii) DMP, p-TSA, CH2Cl2; (iii) OsO4, t-BuOH/H2O (5:2);
(iv) (a) O3, MeOD, -78 °C, (b) NaBH4, 0 °C-rt; (v) (a) O3, MeOD,
-78 °C, (b) NaBD4, 0 °C-rt; (vi) Ac2O, pyridine, CH2Cl2; (vii)
MeOH, HCl (concd), rt.
(1S ,2S )-3-B r o m o c y c lo h e x a -3,5-d ie n e -1,2-d io l-1,2,4,
5,6-d 5 (2b). Perdeuterated diol 2b was obtained from Genencor
International and used without further purification: mp 92-
95 °C; [R]27 +23.8 (c 1.73, CHCl3); IR (KBr) cm-1 3348, 2262,
Sch em e 2a
D
1561, 1406, 1352; 1H NMR (CDCl3) δ 2.26 (bs, OH); 13C NMR
(CDCl3) δ 131.2, 130.9, 130.7, 128.1, 127.9, 127.6, 126.8, 123.7,
123.4, 123.2, 73.6, 73.4, 73.1, 70.8, 70.6, 70.4; MS (CI) m/z (rel
intensity) 197 (M+ + 1)(4 ), 195 (4), 180 (13), 178 (18), 148 (5),
116 (7), 99 (57), 41 (100).
(1S,2S,3S,4S)-5-Ch lor o-3,4-O-isop r op ylid en ecycloh ex-5-
en e-1,2,3,4-tetr ol-1,2,3,4,6-d 5 (4a ). Procedures for acetonide
formation and subsequent diol formation have been previously
described.11e,24 Spectral and physical data for diol 4a : [R]26
D
-33.1 (c 2.07, CHCl3); IR cm-1 3405, 2988, 2936, 2168, 1632,
1373; 1H NMR (CDCl3) δ 2.96 (s, 2OH), 1.41 (s, 3H), 1.38 (s,
3H); 13C NMR (CDCl3) δ 132.4, 126.8, 126.5, 126.3, 110.3, 75.7,
75.5, 75.3, 74.7, 74.4, 74.2, 69.2, 68.9, 68.7, 66.2, 66.0, 65.7, 27.5,
25.9; MS (CI) m/z (rel intensity) 226 (M+ + 1) (100), 210 (30),
150 (65), 103 (32); HRMS calcd for C9D5H9ClO4 226.0894, found
226.0901, error 2.7 ppm.. Anal. Calcd for C9D5H8ClO4: C, 47.90;
H, 5.80. Found: C, 47.70; H, 5.89.
(1S,2S,3S,4S)-5-Br om o-3,4-O-isop r op ylid en ecycloh ex-5-
en e-1,2,3,4-tetr ol-1,2,3,4,6-d 5 (4). Procedures for acetonide
formation and subsequent diol formation have been previously
described.11e, 24 Spectral and physical data for diol 4b: mp 122-
124 °C; [R]27 -8.5 (c 1.00, CHCl3); IR (KBr) cm-1 3507, 3388,
D
a
1
Reagents: (i) O3, MeOH, -78 °C; (ii) NaBH4, 0 °C-rt; (iii)
2986, 2933, 2162, 1629, 1370; H NMR (CDCl3) δ 4.12 (s, OH),
4.04 (s, OH), 1.44 (s, 3H), 1.41 (s, 3H); 13C NMR (CDCl3) δ 130.9,
130.6, 130.5, 123.4, 110.2, 75.9, 75.7, 75.5, 69.1, 68.8, 68.6, 67.0,
66.8, 66.5, 27.6, 26.1; MS (CI) m/z (rel intensity) 270 (M+ + 1)
(100), 254 (27), 194 (5), 103 (57). Anal. Calcd for C9D5H8BrO4:
C, 40.01; H, 4.85. Found: C, 39.92; H, 4.86.
NaBD4, 0 °C-rt; (iv) LAH, THF, rt; (v) LAD, THF, rt.
of the corresponding hydrido derivative22 establishes the
stereochemistry of the methylglycoside to be as depicted.
The syntheses of deuterated mannitols were also
undertaken from deuteriodiol 4b (Scheme 2). Diol 4b
was protected as its disilyl ether 8 and subjected to
ozonolysis and reductive workup. When sodium boro-
hydride was employed as the reducing agent, methyl
(2S,3S,4R,5S)-6-hydroxy-4,5-bis((tert-butyldimethylsily)-
loxy)-2,3-O-isopropylidenehexanoate-2,3,4,5,6-d5 (9) was
the major product; with sodium borodeuteride, methyl
(2S,3S,4R,5S)-6-hydroxy-4,5-bis((tert-butyldimethylsilyl)-
oxy)-2,3-O-isopropylidenehexanoate-1,2,3,4,5,6,6-d6 (10)
was obtained.
(1S,2S,3S,4S)-5-Br om o-1,2-bis((ter t-bu tyld im eth ylsilyl)-
oxy)-3,4-O-isopr opyliden ecycloh ex-5-en e-1,2,3,4,6-d5 (8). Diol
4b (916.2 mg, 3.392 mmol) was dissolved in dry CH2Cl2 (20 mL),
and after 5 min, TBSCl (2.159 g, 4.22 equiv) was added followed
by imidazole (1.1781 g, 5.1 equiv). Stirring was continued for 9
h whereupon more TBSCl (1.0854 g, 2.12 equiv) and imidazole
(522.5 mg, 2.26 equiv) were added. After an additional 23 h 45
min of stirring, the reaction was filtered, concentrated onto silica
gel, and flash chromatographed (19:1 hexane:ethyl acetate) to
yield the title compound as a white solid in 100% yield: [R]27
D
-45.6 (c 1.01, CHCl3); IR (KBr) cm-1 2928, 2891, 2856, 2137,
1627, 1472; 1H NMR (CDCl3) δ 1.42 (s, 3H), 1.39 (s, 3H), 0.91
(s, 9H), 0.87 (s, 9H), 0.097 (s, 6H), 0.089 (s, 3H), 0.084 (s, 3H);
Esters 9 and 10 were reduced with lithium aluminum
hydride (LiAlH4) and lithium aluminum deuteride (Li-
AlD4), respectively, to give the target mannitols: ((tert-
butyldimethylsilyl)oxy)-2,3-O-isopropylidene-D-mannitol-
(23) The LiAlH4 (LiAlD4) reductions also led to the cleavage of one
of the silyl protecting groups, although it is not clear from the
spectroscopic evidence which one.
(24) Hudlicky, T.; Fan, R.; Tsunoda, T.; Luna, H.; Andersen, C.;
Price, J . D. Isr. J . Chem. 1991, 31, 229.
(22) Goux, W. J . Carbohydr. Res. 1988, 184, 47.

Notes
J . Org. Chem., Vol. 61, No. 12, 1996 4153
13C NMR (CDCl3) δ 121.4, 110.0, 27.5, 26.2, 26.0, 25.7, 18.3, 18.1,
-4.4, -4.5, -4.8, -4.9; MS (CI) m/z (rel intensity) 498 (M+ + 1)
(7), 442 (15), 440 (14), 426 (20), 424 (20) 384 (63), 382 (60), 360
(100); HRMS calcd for C21D5H37BrO4Si2 498.2119, found 498.2101,
error 3.6 ppm. Anal. Calcd for C21D5H36BrO4Si2: C, 50.58; H,
8.29. Found: C, 50.67; H, 8.26.
3.69 (d, 1H, J ) 6 Hz), 2.38 (d, OH, J ) 5.6 Hz), 1.58 (s, 3H),
1.36 (s, 3H), 0.91 (s, 9H), 0.90 (s, 9H), 0.121 (s, 3H), 0.118 (s,
3H), 0.099 (s, 3H), 0.094 (s, 3H); 13C NMR (CDCl3) δ 170.8, 110.0,
63.3, 63.1, 62.9, 52.0, 26.4, 26.0, 25.9, 25.6, 18.4, 18.1, -4.3, -4.4,
-4.8, -4.9; MS (CI) m/z (rel intensity) 468 (M+ - 15) (2), 428
(15), 353 (9), 295 (20), 143 (28), 59 (100). Anal. Calcd for
Gen er a l P r oced u r e for Ozon olysis. The compound was
dissolved in methyl alcohol-d and cooled to -78 °C. A stream
of O3/O2 was passed until a dark yellow/green color persisted
for 5 min. After the excess O3 was removed at -78 °C, the
temperature was raised to 0 °C and NaBH4 (NaBD4) was added.
The reaction was brought to ambient temperature, and more
NaBH4 (NaBD4) (equivalents varied) was added until thereduc-
tion was complete. The reaction was diluted with water and
brought to pH ∼2 with HCl (1 N). The solution was then
extracted with ethyl acetate. The combined organic layers were
washed with brine (2×), dried (MgSO4), filtered, and concen-
trated.
Acetyl 4,6-Dia cetyl-2,3-O-isop r op ylid en e-D-m a n n osid e-
2,3,4,5,6-d 5 (5). The crude ozonolysis product (prepared from
97.2 mg of diols 4) was dissolved in excess acetic anhydride and
pyridine and permitted to stir for approximately 12 h whereupon
ethanol was added followed by water. The aqueous layer was
extracted with ethyl acetate (5×). The combined organic layers
were washed with brine, dried, and concentrated onto silica gel
and subjected to flash chromatography (3:2 ether:hexane) to give
a 3:2 epimeric ratio of triacetyl-d5 mannose 5 as a colorless oil
(9 mg, 7%): IR cm-1 2991, 1747, 1458, 1438, 1374; 1H NMR
(CDCl3) δ 6.16 (s, 1H), 4.59 (s, 0.6H), 4.15 (s, 0.4H), 2.08 (s, 6H),
2.06 (s, 3H), 1.47 (s, 3H), 1.30 (s, 3H); 13C NMR (CDCl3) δ 170.6,
169.6, 169.2, 113.5, 100.5, 25.9, 24.9, 21.0, 20.9, 20.8; MS (CI)
m/z (rel intensity) 336 (M+ - 15) (10), 292 (84), 234 (29), 61 (38),
43 (100). Anal. Calcd for C15D5H17O9; C, 51.27; H, 6.31. Found:
C, 51.36; H, 6.27.
C
22D5H41O7Si2: C, 54.61, H, 9.58. Found: C, 54.78, H, 9.59.
Met h yl (2S,3S,4R,5S)-6-H yd r oxy-4,5-b is((ter t-b u t yld i-
m e t h ylsilyl)oxy)-2,3-O-isop r op ylid e n e h e xa n oa t e -2,3,4,
5,6,6-d 6 (10). Ester 10 was prepared from acetonide 8 (211.6
mg, 0.424 mmol) following the general ozonolysis procedure and
was subjected to flash chromatography (17:3 hexane:ethyl
acetate) to yield pure ester 10 as a colorless oil (81.1 mg,
39.4%): [R]22 +12.1 (c 2.09, CHCl3); IR cm-1 3499, 2953, 2930,
D
1
2886, 2857, 2116, 1736, 1472; H NMR (CDCl3) δ 3.73 (s, 3H),
2.31 (bs, OH), 1.56 (s, 3H), 1.34 (s, 3H), 0.89, (s, 9H), 0.87 (s,
9H), 0.099 (s, 3H), 0.096 (s, 3H), 0.077 (s, 3H), 0.072 (s, 3H); 13
C
NMR (CDCl3) δ 170.8, 110.0, 52.0, 26.5, 26.0, 25.9, 25.6, 18.5,
18.1, -4.3, -4.4, -4.8, -4.9; MS (CI) m/z (rel intensity) 485 (M+
+ 1) (16), 469 (18), 453 (10), 427 (100), 353 (27), 295 (45). Anal.
Calcd for C22D6H40O7Si2: C, 54.50; H, 9.56. Found: C, 54.61; H,
9.59.
Gen er a l P r oced u r e for Lith iu m Alu m in u m Hyd r id e/
Lith iu m Alu m in u m Deu ter id e Red u ction s. The ester was
dissolved in freshly distilled THF and cooled to 0 °C. After ∼3
min, LAH (LAD) was added. The reaction was slowly brought
to room temperature and LAH (LAD) was again added (1 equiv).
The reaction was cooled to -78 °C, and the reaction was
quenched with water (1 µL/mg LAH), NaOH (10% aqueous, 1
µL/mg LAH), and water (3 µL/mg LAH). The reaction was
brought to room temperature and stirred until a white paste
formed. The paste was filtered, concentrated onto silica gel, and
flash chromatographed (3:2 ethyl acetate:hexane) to give pure
mannitols.
Acetyl 4,6-Dia cetyl-2,3-O-isop r op ylid en e-D-m a n n osid e-
1,2,3,4,5,6,6-d 7 (6). Compound 6 (331.3 mg, 35%) was prepared
following the above procedures from compound 4a (601.3 mg,
2.7 mmol). 6: [R]26D +22.0 (c 2.06, CHCl3); IR cm-1 2991, 2940,
((ter t-Bu t yld im et h ylsilyl)oxy)-2,3-O-isop r op ylid en e-D-
m a n n itol-2,3,4,5,6-d 5 (11). Mannitol 11 (13.9 mg, 44%) was
prepared from ester 9 (44.9 mg, 0.0928 mmol) as a colorless oil
following the general procedure for LAH reductions. 11: mp
44-47 °C; [R]22D +4.77 (c 1.09, CHCl3); IR cm-1 3403, 2953, 2930,
2886, 2856, 2173, 1472; 1H NMR (CDCl3) δ 3.81 (m, 3H), 3.04
(s, OH), 2.73 (s, OH), 2.69 (bs, OH), 1.51 (s, 3H), 1.40 (s, 3H),
0.90 (s, 9H), 0.091 (s, 6H); 13C NMR (CDCl3) δ 108.1, 64.0, 63.8,
63.6, 61.2, 27.0, 25.8, 24.9, 18.2, -5.4, -5.5; MS (CI) m/z (rel
intensity) 344 (M+ + 3) (4), 343 (M+ + 2) (8), 342 (M+ + 1) (10),
326 (5), 308 (4), 284 (34), 134 (54), 73 (95), 59 (100). Anal. Calcd
for C15D5H27O6Si: C, 52.75; H, 9.45. Found: C, 52.65; H, 9.40.
((ter t-Bu t yld im et h ylsilyl)oxy)-2,3-O-isop r op ylid en e-D-
m a n n itol-1,1,2,3,4,5,6,6-d 8 (12). Mannitol 12 (37 mg, 34%) was
prepared from ester 10 (159.4 mg, 0.3197 mmol) as a colorless
oil following the general procedure for LAD reductions. 12:
1
2188, 1747, 1434; H NMR (CDCl3) δ 2.03 (s, 6H), 2.02 (s, 3H),
1.42 (s, 3H), 1.26 (s, 3H); 13C NMR (CDCl3) δ 170.6, 169.5, 169.1,
113.4, 25.9, 24.8, 20.94, 20.87, 20.7; MS (EI) m/z (rel intensity)
338 (M+ - 15) (40), 294 (10), 266 (12), 174 (18), 43 (100); HRMS
calcd for C14H12D7O9 338.1468, found 338.1468, error 0.0 ppm.
Anal. Calcd for C15H15D7O9: C, 50.97; H, 6.28. Found: C, 50.69;
H, 6.35.
Meth yl 2,3,4,6-Tetr a a cetyl-D-m a n n osid e-1,2,3,4,6,6-d 6 (7).
To the crude ozonolysis product (113.4 mg, 0.5 mmol) in
methanol (1 mL) was added several drops of concentrated
hydrochloric acid. After 24 h the solution was concentrated and
the residue dissolved in excess pyridine and acetic acid. After
5 h the solution was concentrated and the residue flash chro-
matographed (2:1 hexane:ethyl acetate) to yield the title com-
pound (42.4 mg, 23%) which exhibited spectral properties similar
to those described previously for nondeuterated analog:22 1H
NMR (CDCl3) 3.41 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H), 2.04 (s,
3H), 1.99 (s, 3H).
[R]25 +2.70 (c 2.04, CHCl3); IR cm-1 3440, 2954, 2930, 2884,
D
2857, 2101, 1471; 1H NMR (CDCl3) δ 3.06 (s, OH), 2.74 (s, OH),
2.70 (s, OH), 1.51 (s, 3H), 1.40 (s, 3H), 0.91 (s, 9H), 0.095 (s,
3H), 0.092 (s, 3H); 13C NMR (CDCl3) δ 108.1, 27.0, 25.8, 24.9
18.2, -5.4, -5.5; MS (CI) m/z (rel intensity) 346 (M+ + 2) (2),
345 (M+ + 1) (5), 344 (M+) (1), 287 (8), 250 (4), 155 (10), 137
(13), 73 (40), 59 (72), 41 (100). Anal. Calcd for C15D8H24O6Si:
C, 52.29, H, 9.36. Found: C, 52.04; H, 9.28.
Met h yl (2S,3S,4R,5S)-6-H yd r oxy-4,5-b is((ter t-b u t yld i-
m e t h ylsilyl)oxy)-2,3-O-isop r op ylid e n e h e xa n oa t e -2,3,4,
5,6-d 5 (9). Crude ester 9 was prepared from acetonide 8 (459.6
mg, 0.9217 mmol) according to the general ozonolysis procedure
and was subjected to flash chromatography (17:3 hexane:ethyl
acetate) to yield pure ester 9 as a colorless oil (78.6 mg, 67%
Ack n ow led gm en t. We thank the National Science
Foundation and the University of Florida for the fund-
ing of this work (CHE-9315684 and CHE-9521489).
yield): [R]23.5 +12.9 (c 1.68, CHCl3); IR cm-1 3504, 2954, 2930,
D
1
2881, 2856, 2136, 1735, 1473; H NMR (CDCl3) δ 3.75 (s, 3H),
J O951666S