1-Aza-sugars from D-Glucose. Preparation of 1-Deoxy-5-dehydroxymethyl-Nojirimycin, its analogues and evaluation of glycosidase inhibitory activity

Article abstract of DOI:10.1016/S0968-0896(02)00073-1

D-Glucose derived pentodialdoses 11a-c on reduction followed by tosylation, azide displacement, hydrogenation and protection with -Cbz group gave N-Cbz protected compounds 14a-c, respectively, which on removal of 1,2-acetonide functionality and hydrogenat

Full text of DOI:10.1016/S0968-0896(02)00073-1

Bioorganic & Medicinal Chemistry10 (2002) 2155–2160
1-Aza-sugars from D-Glucose. Preparation of
1-Deoxy-5-dehydroxymethyl-Nojirimycin, Its Analogues
and Evaluation of Glycosidase Inhibitory Activity
Nitin T. Patil,a Sheeja John,b Sushma G. Sabharwalb and Dilip D. Dhavalea,*
aDepartment of Chemistry, Garware Research Centre, University of Pune, Pune-411 007, India
bDepartment of Chemistry, Division of Biochemistry, University of Pune, Pune-411 007, India
Received 11 January2002; accepted 9 February2002
Abstract—d-Glucose derived pentodialdoses 11a–c on reduction followed by tosylation, azide displacement, hydrogenation and
protection with -Cbz group gave N-Cbz protected compounds 14a–c, respectively, which on removal of 1,2-acetonide functionality
and hydrogenation afforded corresponding 1-aza-sugars 3, 9 and 10 in good overall yields. The glycosidase inhibition activity of
these 1-aza-sugars was tested with sweet almond as a rich source of different glycosidases. # 2002 Elsevier Science Ltd. All rights
reserved.
In the last two decades, glycosidase inhibitors such as
aza-sugars have been attractive target molecules for
synthetic as well as bio-chemists not only because they
serve as a useful tool for studying the biological func-
tion of oligosachorides1 but also because theymayhave
great potential as drugs in the treatment of a varietyof
carbohydrate mediated diseases.2 Amongst these aza-
sugars, nojirimycin (1) and 1-deoxynojirimycin ( 2) are
the first naturallyoccurring alkaloids with promising
biological activity.3 In recent years, improved glycosi-
dase inhibition is being examined for each hydroxyl
substituent in 1 or 2 and systematic data in the inhibi-
tion of b-glucosidases is documented in the literature.4
In this aspect, it has been noted that the removal of the
hydroxymethyl substituent at C-5 in 2 had verylittle
effect on enzyme substrate activity.5 In view of this,
Ganem et al. have first reported the synthesis of piper-
idine triols 3, 4 and 5 and referred these compounds as
1-aza-sugars or 1-N-imino sugars wherein the nitrogen
atom is considered to be at one position.5 Later on,
Genjiro and coworkers have isolated 1-aza-sugars 3 and
4 from Eupatorium fortunei Turz.6 Since then a number
of new derivatives of 5-de(hydroxymethyl)-1-deoxy-
nojirimycin 3 with different stereochemical orientation
of the –OH functionalityat C-3/C-4/C-5 (e.g., 3, 4 and 5)
as well as the replacement of one of the –OH functionality
at C-3/C-5 with hydroxymethyl substituent (e.g., 6, 7
and 8) have been synthesised and evaluated for glycosi-
dase inhibition.7 As a part of our continuing interest in
the synthesis of nojirimycin analogues,8 we are now
reporting an efficient route for the synthesis of 3S, 4R,
5R piperidine triol (3), 3S, 4S, 5R piperidine triol (9),
and 3S,4S(2-hydroxyethyl), 5R piperidine triol (10) and
discuss their glycosidase inhibitory activity.
Results and Discussion
Synthesis of 3S, 4R, 5R piperidine triol (3) and 3S, 4S,
5R piperidine triol (9)
As shown in Scheme 1, d-glucose was converted to
1,2-O-isopropylidene-3-O-benzyl-a-d-xylo-pentodialdose
11a in 68% overall yield as reported earlier by us.8 The
pentodialdose 11a was reduced with sodium borohydride
in methanol to give an alcohol which on tosylation
afforded 5-O-tosyl 1,2-O-isopropylidene-3-O-benzyl
a-d-xylofuranose 12a in good yield. The nucleophilic
displacement of the tosyl group by NaN3 in DMF led to
the formation of an azido compound 13a.9 Subse-
quently, 13a was subjected to catalytic hydrogenation
and the amino alcohol thus obtained was directly
treated with benzylchloroformate in the presence of
sodium bicarbonate in ethanol to give 5-(N-benzoxy-
* Corresponding author. Tel.:+91-20-569-6061; fax.:+91-20-569-
1728; e-mail: ddd@chem.unipune.ernet.in
0968-0896/02/$ - see front matter # 2002 Elsevier Science Ltd. All rights reserved.
PII: S0968-0896(02)00073-1

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Nitin T Patil et al. / Bioorg. Med. Chem. 10 (2002) 2155–2160
Scheme 1. Reagents and conditions: (i) NaBH4, MeOH, rt, 15 min; (ii) TsCl, pyridine, rt, 12 h; (iii) NaN3, DMSO, 100 ꢀC, 6 h; (iv) 10% Pd/C, H2,
80 psi, MeOH, 12 h; (v) CbzCl, NaHCO3, EtOH-H2O (8:2), rt, 2 h; (vi) TFA-H2O (3:2), rt, 2 h; (vii) 10% Pd/C, H2, 8 psi, MeOH, 12 h.
carbonylamino)-5-deoxy 1,2-O-isopropylidene-3-O-ben-
zyl-a-d-xylofuranose 14a. Treatment of 14a with TFA-
water followed by catalytic hydrogenation afforded 3.
The spectral and analytical data of 3 was found to be in
good agreement with that reported.5 The same sequence
of reactions was repeated using d-glucose derived 1,2-O-
isopropylidene-3-O-benzyl-a-d-ribo-pentodialdose
11b
and 3S, 4S, 5R piperidine triol 9 was synthesised in
overall 62% yield. Compound 9 was treated with
.
MeOH-HCl and amine-hydrochloride salt 9 HCl was
isolated as a solid. The spectral and analytical data of
.
9 HCl was in consonance with that reported [mp=162–
163 ꢀC (reported mp=161–163 ꢀC)].7l
Synthesis of 3S, 4S(2-hydroxyethyl), 5R piperidine triol
(10):
Figure 1.
The literature surveyindicates that different analogues
of 3 wherein –OH group at C-3 and C-5 is replaced
with a hydroxymethyl group are known. However,
substitution of C-4 hydroxy with either ÀCH2OH or
ÀCH2CH2OH group is not reported so far. In the pre-
sent study, we thought of synthesising a new analogue
wherein C-4 hydroxyl group in 3 is substituted by
ÀCH2CH2OH functionalitywith inversion of orienta-
tion of the group. Thus, d-glucose was converted to
Conformational Assignment of 3, 9 and 10
Nojirimycin (1) and 1-deoxy-nojirimycin (2) are known
to exist in 4C1 conformation. Recently,8d we have
described the conformational studyof 1-deoxy- d-gluco-
homonojirimycin 15a and 1-deoxy-l-ido-homonojiri-
mycin 15b and shown that both the compounds exist in
4C1 conformations although, in 15b the bulky
ÀCH2CH2OH group at C-5 is axiallyoriented (Fig. 2).
The assignment of conformation is based on the 1H
NMR spectra and coupling constant information
between H-1, H-2, H-3, H-4 and H-5. In the case of 1-
aza-sugars 3, 9 and 10, we have observed a dramatic
known aldehyde 11c with a-oriented –CH2CH2OH
10
functionalityat C-3 bya known method.
The alde-
hyde group in 11c was subjected to NaBH4 reduction in
methanol and the product thus obtained on tosylation
and nucleophilic displacement with NaN3 afforded
azido compound 13c (Scheme 1). Catalytic hydrogena-
tion of 13c followed byCbz protection afforded 14c as a
thick oil in 90% yield. Treatment of 14c with TFA-H2O
and catalytic hydrogenation afforded a compound that
on purification with amberlite IR 400A (OHÀ) resin
gave a piperidine triol 10 in 95% yield.
1
differences in the H NMR spectra which we would like
to attribute to the conformational changes of these
molecules. Thus, in case of compound 3 two conforma-
5
2
tions C2 (3a) and C5 (3b) are possible. In conforma-
tion 3a, it is expected that the coupling constant J2a,3a
,
J3a,4a and J4a,5a should be large (Jaa=7–9 Hz) while
in the case of 3b coupling constants should be small as

Nitin T Patil et al. / Bioorg. Med. Chem. 10 (2002) 2155–2160
2157
Figure 2. Different conformations of 3, 9 and 10.
Biological Activity
Table 1. Inhibitorypotencies of 1-aza-sugars (IC 50, CM) of 3, 9 and
10
Almond seeds are known to be a rich source of glycosi-
dases namely a-glucosidases (E.C. 3.2.1.23), b-gluco-
sidases (E.C. 3.2.1.2.4), a-galactosidases (E.C.
3.2.1.2.2), b-galactosidases (E.C. 3.2.1.2.3), and a-man-
nosidases (E.C. 3.2.1.2.4). Therefore, the potencyof 1-
aza-sugars 3, 9 and 10 as glycosidase inhibitors was
evaluated using almond seed extract as a glycosidase
source. The IC50 values obtained are summarised in
Table 1.
Enzyme
3
9
10
b-Glucosidase
a-Glucosidase
b-Galactosidase
a-Galactosidase
a-Mannosidase
1.40Â10À3
—
3.6Â10À6
4.44Â10À6
—
3.18Â10À6
4.97Â10À6
—
3.27Â10À6
—
—
—
—
—
H-3, H-4 and H-5 are equatoriallyoriented. In 3, the
appearance of triplet at 3.37 (J3,4=8.9 Hz), a doublet of
doublet at 3.32 (J2e,2a=12.5, J2e,3a=4.8 Hz), and
another doublet of doublet at 2.80 (J2e,2a=12.7,
J2a,3a=12.2 Hz) clearlyindicated the trans-diaxial rela-
tionship between the H-2a, H-3a, and H-3a, H-4a con-
In the case of 3, the observed IC50 value for b-gluco-
sidase inhibition is found to be comparable to that
reported.5 The glycosidases inhibition activity of 9,
.
although known in the literature as 9 HCl, is not
reported. Our results demonstrate that 9 is a potent
b-glucosidase inhibitor than 3. This indicates that
inversion of configuration at C-4 in piperidine triol,
with –OH substituent, has a significant increase in
b-glucosidase inhibition activity. Biological activities of
3 and 10 (wherein b-hydroxy functionality at C-4 was
replaced by a-CH2CH2OH functionality) were com-
pared. The comparative studydemonstrates that the
replacement of C4-OH with a-oriented –CH2CH2OH
functionalityresulted in increased potencytowards
b-glucosidases with high specificity.
5
firming C2 conformation 3a. However, in the case of
1
compound 9, the H NMR showed small coupling con-
stant values (WH=5 Hz) for H-2, H-3 and H-3, H-4
protons indicating 2C5 conformation 9b. In the 1H
NMR spectra of 10, two broad doublets at 2.50 and
2.74 (J2e,2a=13.5 Hz) corresponding to protons H-2a
and H-2e suggested that H-3 is equatoriallyoriented
and bisecting the H-2 protons (dihedral angle ꢁ55ꢀ).
The H-3 and H-5 protons also appeared as narrow
2
multiplets thus indicating C5 conformation 10b. The
conformation 9b and 10b is stabilised byintramolecular
hydrogen bonding (Fig. 2). In addition, in the case of
10b, the CH2CH2OH is equatorial while in 10a it is
axial. This further stabilises 10b in relation to 10a.
In conclusion, we have demonstrated an efficient and
straightforward methodologyfor the synthesis of 1-aza-
sugars that can be reproduced on multigram scale. The
concept that we have demonstrated herein clearly

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Nitin T Patil et al. / Bioorg. Med. Chem. 10 (2002) 2155–2160
indicates a design strategyfor the development of new
analogues of 1-aza-sugars. The synthesised compounds
were tested for inhibition of glycosidases and are found
to be good b-glucosidase inhibitors.
tosyl chloride (0.597 g, 3.13 mmol) was added. The
reaction mixture was warmed to room temperature and
stirred for 12 h. Water was added and resulting reaction
mixture was extracted with ethyl acetate (10 mLÂ3).
The organic layer was washed with cooled dil HCl,
water and dried. Evaporation of organic layer afforded
tosyl derivative 12c (1.02 g, 85%) as a thick liquid.
Experimental
1H NMR (300 MHz) and 13C NMR (75 MHz) spectra
were recorded using CDCl3 as a solvent unless other-
wise stated. Chemical shifts are reported in ppm (d)
Rf=0.65 (n-hexane/ethyl acetate=7/3); [a]D=+21.5 (c,
0.6, CHCl3); IR nmax (neat)=1656; H NMR (CDCl3,
1
300 MHz): 1.35 (3H, s, CH3), 1.51 (3H, s, CH3), 1.64–
1.80 (1H,m, CH2/H-3), 1.88–2.00 (1H, m, CH2/H-3),
2.10–2.22 (1H, m, CH2/H-3), 2.49 (3H, m, CH3), 3.58–
3.70 (2H, m, CH2OBn), 3.98–4.06 (1H, m, H-4), 4.14
(1H, dd, J=4.2, 11.1 Hz, H-5), 4.32 (1H, dd, J=2.7,
11.1 Hz, H-5), 4.7 (2H, bs, O-CH2Ph), 4.62 (1H, dd,
J=3.9, 4.2 Hz, H-2), 5.74 (1H, d, J=3.9 Hz, H-1),
7.2 (7H, Ar–H), 7.85 (2H, d, J=8 Hz, Ar–H); 13C
NMR (CDCl3, 75 MHz): 25.0 (CH3), 26.2 (CH3), 40.8
(CH2), 42.9 (CH2), 51.7 (O-CH2), 67.2 (O-CH2Ph),
73.9, 75.5, 81.8 (C-2, C-3, C-4), 104.5 (C-1), 111.3 (O–
C–O), 127.6, 128.4, 129, 138.3, 144 (Ar–C); Anal. calcd
for C24H30SO7: C, 62.87; H, 6.60. Found C, 62.90; H,
6.80.
relative to internal standard Me4Si. IR spectra (n, cmÀ1
)
were measured as thin films or Nujol mulls or KBr pel-
lets. Optical rotations were recorded at 25 ꢀC. Whenever
required the reactions were carried out in oven-dried
glassware under dryN2. On workup, reaction mixture
was extracted with organic solvents, evaporated at
reduced pressure with rotaryevaporator. Thin laeyr
chromatographywas performed on 0.25 mm pre-
coated silica gel, flash chromatographywas carried out
on silica gel 200–400 mesh and column chromatography
on silica gel 100–200 mesh. The organic solvents such as
n-hexane, THF, diethyl ether, chloroform, petroleum
ether (pet. ether, 60–70 ꢀC fraction), ethyl acetate and
methanol were purified and dried before use. LAH,
Cbz-Cl were purchased from Aldrich and/or Fluka. 1,2-
O-Isopropylidene-3-[2-benzyloxyethyl]-a-D-ribo-pento-
dialdo-1,4-furanose 11c and azido compounds 13a,b
were prepared as per literature procedure and char-
acterisedbyspectral methods.
1,2-O-Isopropylidene-3(2-benyloxyethyl)-5-azido-5-deoxy-
ꢀ-D- ribofuranose (13c). A mixture of 12c (0.6 g, 1.298
mmol) and NaN3 (0.42 g, 6.49 mmol) in anhydrous
DMF was stirred at 100 ꢀC for 6 h. The reaction mix-
ture was cooled to room temperature. After adding
water the reaction mixture was extracted with ethyl
acetate. The organic layer was dried and concentrated
to furnish azido compound 13c (0.37 g, 86%) as a thick
liquid.
Assay method
The substrates (2 mM in 0.025 M citrate buffer, pH 4.0)
used were p-nitrophenyl N-acetyl-b-glucosaminide,
p-nitrophenyl b-d-galactoside, p-nitrophenyl a-d-galacto-
side, p-nitrophenyl a-d-glucoside, p-nitrophenyl a-d-
mannoside, respectively. The test compound was
pre-incubated with the enzyme (almond seed extract)
for 1 h at 37 ꢀC. The enzyme reaction was initiated
bythe addition of 100 mL substrate. Controls were
run simultaneouslyin the absence of the test com-
pound. The reaction was terminated at the end of
90 min bythe addition of borate buffer (0.05 M,
pH=9.8) and absorbance of the liberated p-nitrophenol
was measured at 420 nm. The IC50 values were
determined (IC50 value is the concentration of inhibitor
at 50% ofenzyme activity). One unit of glycosidase
activityis defined as the amount of enzmye that
hydrolysed 1 mmole of p-nitrophenyl pyranoside per
min at 25 ꢀC.
Rf=0.72 (n-hexane/ethyl acetate=7/3); [a]D=+31.00
(c, 0.6, CHCl3); IR ꢀmax (neat)=2102; 1H NMR
(CDCl3, 300MHz): 1.32 (3H, s, CH3), 1.54–2.02 (3H, m,
CH2 and H-3), 3.48 (2H, narrow multiplet, N3CH2),
3.65–3.78 (2H, m, O–CH2), 3.80–3.85 (1H, m, H-4), 4.67
(1H, dd, J=3.6, 3.5 Hz, H-2), 5.11 (2H, ABq, J=12.3
Hz, CH2Ph), 5.76 (1H, d, J=3.5 Hz, H-1), 7.2 (5H, s,
Ar–H); 13C NMR (CDCl3, 75 MHz): 24.9 (CH3), 26.3
(CH3), 26.8 (CH2), 42.6 (CH2), 51.7 (OCH2), 68.2
(CH2Ph), 72.9, 80.5, 81.2 (C-2, C-3, C-4), 104.8 (C-1),
111.7 (O–C–O), 127.6, 128.4, 138.3 (Ar–C); Anal. calcd
for C17H23N3O4: C, 61.24; H, 6.95. Found C, 61.34; H,
7.01.
General procedure for hydrogenation and N-Cbz protec-
tion. A mixture of azido compounds (1 mmol) and
10% Pd/C (10% w/w of the starting) in methanol was
subjected to hydrogenation at 80 psi for 12 h. The cat-
alyst was filtered through Celite and washed with
methanol (5 mL). To the filtrate, sodium bicarbonate (4
mmol) and benzyloxycarbonyl chloride (1.2 mmol) was
added at 0 ꢀC. The mixture was stirred at 25 ꢀC for 1 h.
The methanol was evaporated and water (2 mL) was
added. The reaction mixture was extracted with chloro-
form (3Â10 mL), washed with water, dried and the
organic layer was evaporated to give a thick liquid.
Purification bycolumn chromatographyafforded N-Cbz
compound.
1,2-O-Isopropylidene-3(2-benyloxyethyl)-5-O-tosyl-ꢀ-D-
ribofuranose (12c). To an ice cooled solution of 11c (0.8
g, 2.61 mmol) in methanol–water (4:1, 10 mL) was
added NaBH4 (0.118 g, 3.13 mmol) over a period of 10
min. The reaction mixture was warmed to room tem-
perature and stirred for 0.5 h. Methanol was evaporated
and resulting reaction mixture was extracted with ethyl
acetate (10 mLÂ3). The organic layer was dried over
sodium sulphate and evaporated to obtain product as a
thick liquid. The product thus obtained was dissolved in
pyridine (1.05 mL, 13.05 mmol), cooled to 0 ꢀC and

Nitin T Patil et al. / Bioorg. Med. Chem. 10 (2002) 2155–2160
2159
1,2-O-Isopropylidene-3-O-benzyl-5-(N-benzoxycarbonyl-
amino)-5-deoxy-ꢀ-D-xylofuranose (14a). White solid;
mp=119–121 ꢀC; yield=95%; Rf=0.36 (n-hexane/ethyl
acetate=7/3); [a]D=31.25 (c, 0.8, CHCl3);IR ꢀmax
(neat)=1687, 3350–3150 (broad band); 1H NMR
(CDCl3 + D2O, 300 MHz): 1.23 (3H, s, CH3), 1.45 (3H,
s, CH3), 3.20–3.25 (1H, dd, J=3.3, 14.0 Hz), 3.60–3.68
(1H, dd, J=9.3, 14.0 Hz), 4.0–4.06 (2H, m, H-3, H-4),
4.60 (1H, d, J=3.6 Hz, H-2), 5.05 (2H, s, O-CH2), 5.85
(1H, d, J=3.6 Hz, H-1), 7.17–7.22 (5H, m, Ar–H); 13C
NMR (CDCl3 + D2O, 75 MHz): 26.1 (CH3), 26.8
(CH3), 38.3 (N–CH2), 67.5 (O–CH2), 73.9, 80.1, 84.9
(C-2/C-3/C-4), 104.7 (C-1), 111.5 (O–C–O), 128.16,
128.4, 128.6, 135.8 (Ar–C), 157.9 (NCO); Anal. calcd
for C16H21NO6: C, 59.43; H, 6.55. Found C, 59.60; H,
6.60.
dissolved in methanol and stirred with basic ion-
exchange resin IRA 400 for 10 min. Filtration through
Celite and concentration of the methanol under reduced
pressure afforded 1-aza-sugar.
(3S, 4R, 5R) 3,4,5-Piperidine triol (3). Semi solid;
yield=92%; Rf=0.18 (chloroform/methanol=1/1);
[a]D=0.0 (c 0.9, MeOH); IR nmax (KBr pellet)=3300–
3250 (broad band); 1H NMR (CDCl3+D2O, 300 MHz):
2.80 (1H, dd, J=12.8, 12.3 Hz, H2a/H6a) 3.32 (1H, dd,
J=12.8, 4.8 Hz, H-2e/H-6e), 3.37 (1H, t, J=8.95 Hz,
H-4), 3.64 (1H, m, H-3/H-5); 13C NMR (CDCl3
+
D2O, 75 MHz): 46.3 (C-2, C-6), 66.9 (C-3, C-5), 74.76
(C-4); Anal. calcd for C5H11NO3: C, 45.10; H, 8.33.
Found C, 45.32; H, 8.55.
(3S, 4S, 5R) 3,4,5-piperidine triol (9). Semi solid;
yield=94%; Rf=0.18 (chloroform/methanol=1/1);
[a]D=0.0 (c 0.8, MeOH); IR nmax (KBr pellet)=3300–
3200 (broad band); 1H NMR (CDCl3 + D2O,
300 MHz): 3.0–3.04 (4H, m, H-2, H-6), 3.81–3.85 (3H,
m, H-3,H-4, H-5); 13C NMR (CDCl3 + D2O, 75 MHz):
44.29 (C-2, C-6), 65.55 (C-3, C-5), 68.35 (C-4); Anal.
calcd for C5H11NO3: C, 45.10; H, 8.33. Found C, 45.20;
H, 8.50.
1,2-O-Isopropylidene-3-O-benzyl-5-(N-benzoxycarbonyl-
amino)-5-deoxy-ꢀ-D-ribofuranose (14b). Thick liquid;
yield=93%; Rf=0.18 (n-hexane/ethyl acetate=7/3);
[a]D=25.33 (c, 0.9, CHCl3); IR ꢀmax (neat)=1685,
3360–3100 (broad band); 1H NMR (CDCl3 +D2O,
300 MHz): 1.36 (3H, s, CH3), 1.56 (3H, s, CH3), 3.47–
3.6 (2H, m, N–CH2), 3.7 (1H, dd, J=5.1, 8.8 Hz, H-3),
3.85 (1H, m, H-4), 4.57 (1H, dd, J=3.6, 5.1 Hz, H-2),
5.1 (2H, bs, COCH2), 7.75 (d, J=3.6 Hz, H-1); 13C
NMR (CDCl3 +D2O, 75 MHz): 26.31 (CH3), 26.34
(CH3), 41.1 (N-CH2), 66.88 (O-CH2), 72.50, 78.58,
78.64 (C-2/C-3/C-4), 103. 62 (C-1), 112.72 (O–C–O),
128.08, 128.43, 136.21 (Ar–C), 156.9 NCO); Anal. calcd
for C16H21NO6: C, 59.43; H, 6.55. Found C, 59.53; H,
6.72.
To a stirred solution of compound 9 (0.2 g, 1.50 mmol)
.
in drymethanol was added HCl (0.1 mL) under dryN
The reaction mixture was stirred at room temperature
for 2 h. After 2 h, the solvent was evaporated under
2
.
reduced pressure to give 9 HCl (0.23 g, 95%) as crystal
which was crystallized from DMF–EtOH.
1,2-O-Isopropylidene-3(2-benzyloxyethyl)-5-(N-benzoxy-
carbonylamino)-5-deoxy-ꢀ-D-ribofuranose (14c). White
solid; yield=90%; Rf=0.25 (n-hexane/ethyl acetate=7/
3); [a]D=+31.00 (c, 0.6, CHCl3); IR ꢀmax
(neat)=1680, 3400–3100 (broad band); 1H NMR
(CDCl3 +D2O, 300 MHz): 1.31 (3H, s, CH3), 1.49 (3H,
s, CH3), 1.56–1.7 (1H, m, CH2), 1.82–1.95 (1H, m,
CH2), 2.08–2.19 (1H, m, N-CH2), 2.22 (1H, dd, J=4.8,
12.9 Hz, N-CH2), 3.56–3.64 (3H, m, H-3, O-CH2), 3.95–
4.01 (1H, m, H-4), 4.6 (1H, t, J=4.2 Hz, H-2), 5.8 (1H,
d, J=4.2 Hz, H-1); 13C NMR (CDCl3 + D2O,
75 MHz): 26.3 (CH3), 26.7 (CH3), 27.7 (CH2), 41.4 (N-
CH2/O-CH2) 41.6 (N-CH2/O-CH2), 60.7 (COCH2),
60.7, 80.6, 81.4 (C-2, C-3, C-4), 104.8 (C-1), 111.6 (O–
C–O), 128.2, 128.5, 136.2 (Ar–C), 157.1 (NCO); Anal.
calcd for C18H25NO6: C, 61.52; H, 7.17. Found C,
61.54; H, 7.21.
White solid; mp=162–163 ꢀC (reported mp=161–
163 ꢀC); yield=95%; [a]D=0.0 (c 0.8, MeOH); IR ꢀmax
(KBr pellet)=3300–3200 (broad band); 1H NMR
(CDCl3 +D2O, 300 MHz): 3.12–3.31 (4H, m, H-2, H-
6), 4.05–4.10 (3H, m, H-3, H-4, H-5); 13C NMR (CDCl3
+ D2O, 75 MHz): 44.20 (C-2, C-6), 65.45 (C-3, C-5),
69.30 (C-4).
3S, 4S (2-Hydroxyethyl), 5R piperidine triol (10). Semi
solid; yield=95%; Rf=0.15 (chloroform/methanol=1/
1); [a]D=0.0 (c 0.1, MeOH); IR ꢀmax (KBr pel-
let)=3300–3260 (broad band); 1H NMR (D2O,
300 MHz): 1.05–1.30 (3H, m, CH2, H-4), 2.50 (2H, bd,
J=13.5 Hz, H-2a, H-6a, 2.74 (2H, d, J=13.5 Hz, H-2e,
H-6e), 2.90–3.10 (2H, m, CH2), 3.4 (2H, bs, H-3, H-5);
13C NMR (D2O, 75 MHz): 30.0 (CH2), 36.3 (C-1/CH2),
49.8 (C-1/CH2), 58.6 (C-4), 65.30 (C-3); Anal. calcd
for C7H15NO3: C, 52.15; H, 9.38. Found C, 52.20; H,
9.49.
General procedure for acid hydrolysis and hydrogena-
tion. A solution of N-Cbz compound (1 mmol) in
TFA-H2O (3/2, 2 mL) was stirred at 25 ꢀC for 2 h. Tri-
fluroacetic acid was co-evaporated with benzene to fur-
nish a thick liquid, which was directlyused in the next
reaction. To a solution of the above product in metha-
nol was added 10% Pd/C (10% w/w of the starting) and
solution was hydrogenated at 80 psi for 12 h. The cata-
lyst was filtered through Celite, washed with methanol
and the filtrate concentrated to get crude compound.
The crude compound was washed with chloroform
(which was discarded) and thick oil thus obtained was
Acknowledgements
We are grateful to BRNS, Mumbai for the financial-
support (98/37/11/BRNS). NTP is thankful to CSIR,
New Delhi for the JRF and SRF. We are indebted to
Prof. M. S. Wadia and Prof. N. S. Narasimhan for the
discussion. We gratefullyacknowledge UGC, New
Delhi, for the grant to procure a high field NMR facility
(300 MHz).

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Nitin T Patil et al. / Bioorg. Med. Chem. 10 (2002) 2155–2160
(b) Liang, X.; Lohse, A.; Bols, M. J. Org. Chem. 2000, 65,
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