Probing the aglycon binding site of a β-glucosidase: A collection of C-1-modified 2,5-dideoxy-2,5-imino-D-mannitol derivatives and their structure-activity relationships as competitive inhibitors

Article abstract of DOI:10.1016/j.bmc.2004.04.037

A range of new C-1 modified derivatives of the powerful glucosidase inhibitor 2,5-dideoxy-2,5-imino-D-mannitol has been synthesised and their biological activities probed with the β-glucosidase from Agrobacterium sp. K_i values are compared with

Full text of DOI:10.1016/j.bmc.2004.04.037

Bioorganic & Medicinal Chemistry 12 (2004) 3485–3495
Probing the aglycon binding site of a b-glucosidase: a collection of
C-1-modified 2,5-dideoxy-2,5-imino-
their structure–activity relationships as competitive inhibitors
D
-mannitol derivatives and
a
Tanja M. Wrodnigg, Frederik Diness, Christoph Gruber, Herwig Hausler, Inge Lundt,
b
a
a
b
€
Karen Rupitz, Andreas J. Steiner, Arnold E. Stutz, Chris A. Tarling,
c
a
a,
c
*
€
Stephen G. Withers and Heidrun Wolfler
c
a
€
aGlycogroup, Institut f€ur Organische Chemie, Technische Universit€at Graz, Stremayrgasse 16, A-8010 Graz, Austria
bLaboratory of Organic Chemistry, Department of Chemistry, Technical University of Denmark,
Building 201, DK-2800 Lyngby, Denmark
cDepartment of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada V6T 1Z1
Received 27 February 2004; accepted 27 April 2004
Dedicated to Professor Christian Pedersen
Abstract—A range of new C-1 modified derivatives of the powerful glucosidase inhibitor 2,5-dideoxy-2,5-imino-D-mannitol has
been synthesised and their biological activities probed with the b-glucosidase from Agrobacterium sp. Ki values are compared with
those of previously prepared close relatives. Findings suggest dramatic effects exerted by the aglycon binding site on substrate/
inhibitor binding.
ꢀ 2004 Elsevier Ltd. All rights reserved.
1. Introduction
This fact was attributed to the relatively flat five-mem-
bered ring, which was assumed to more closely mimic
the putative transition state of enzymatic glycoside
hydrolysis, along with its C2-axis of symmetry. Standard
modifications at C-1, such as in the 1-deoxyfluoro (3) as
well as the 1-O-methyl (4) derivatives caused losses of
inhibitory activities in the range of two orders of mag-
nitude (Fig. 2).3
The iminoalditol 2,5-dideoxy-2,5-imino-D-mannitol
(DMDP, 1), a natural product,1 is known as a potent
reversible inhibitor of
rivalling the activity of 1-deoxynojirimycin (1,5-dideoxy-
1,5-imino- -glucitol, DNJ, 2), the paradigmatic -glu-
D-glucosidases and invertase
D
D
cosidase inhibitor in the class of compounds under
consideration (Fig. 1).2
The same was true for 5, the 1-aminodeoxy derivative of
compound 1, which was prepared via an Amadori
rearrangement reaction of 5-azidodeoxy-D-glucofura-
nose.4
H
H
N
N
HO
HO
OH
C-1 modified derivatives with extended alkyl chains such
as compounds 6 as well as 8 and 9, initially designed for
immobilisation studies,5 were synthesised and found to
exhibit inhibitory properties comparable to or even
better than those of parent compound 1.5 Similar results
have been found with 10 and, by other workers,6 with
compound 7 as well as natural products featuring the
iminomannitol basic structure such as the broussone-
tines,7 for example, compound 12. These results sug-
gested that the aglycon binding site is able to exert
HO
OH
HO
OH
OH
2
1
Figure 1. Structures of DMDP (1) and 1-DNJ (2).
Keywords: b-Glucosidase inhibitors; Iminosugars.
* Corresponding author. Tel.: +43-316-873-8747; fax: +43-316-873-
8740; e-mail: stuetz@orgc.tu-graz.ac.at
0968-0896/$ - see front matter ꢀ 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2004.04.037

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T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
H
N
3: X = F
4: X = OMe
7: X = NHAc
8: X = O(CH2)6OH
HO
X
5: X = NH2
9: X = NHCO(CH2)5OH
6: X = NH(CH2)6OH 10: X = NHCO(CH2)4CH3
11: X = NHCO(CH2)2Ph
HO
OH
Figure 2. Structures of compounds 3–11.
binding forces compensating for activity losses caused
by the removal of one crucial hydroxyl group as well as
the loss of symmetry resulting from this formal ‘deoxy-
genation’ (Fig. 3).
Unfortunately and despite all efforts made thus far, for
the b-glucosidase from Agrobacterium sp. there is no
XRD structure available, as yet.
Consequently, we continued to resort to the conven-
tional means of exploration taking advantage of pieces
of information provided by structure–activity consider-
ations based on results with selected new inhibitors.
Further studies revealed that mixed aliphatic–aromatic
side chains exhibit similar properties, for example in
compound 11.8 The general significance of long aliphatic
substituents had also been shown by Legler and co-
workers9 with compound 13 as well as by Vasella and
his group10 with compound 14 in the iminosugar field
and by Ogawa et al.11 for examples in the aminocyclitol
family of compounds such as 15, just to mention a few
relevant examples. To gain a better understanding of the
structure-activity relationships applying in ‘our’ family
of compounds, we have extended the range of deriva-
tives by substitution with markedly lipophilic aromatic
side chains such as in 16 and 17.8 Some of these deriv-
atives, in particular, compounds 18 and 19 featuring
dialkylamino substituents in the respective aromatic
system, exhibited Ki values in the low nanomolar range
exceeding the parent compound’s activity by two orders
of magnitude (Fig. 4).12
In addition, novel chain-extended 1-O-alkyl derivatives
of 2,5-dideoxy-2,5-imino-D-glucitol, the epimer at C-2
of compound 1, were synthesised and their activities
compared with the parent compounds.
2. Results and discussion
To address the respective influence of various parame-
ters such as the nature of the hetero atom linking side
chain and sugar, the nature and type of hybridisation of
the attached carbon or heteroatom, the size of the aro-
matic system as well as the presence and positioning of
the aromatic amine substituent, derivatives 20–27 were
synthesised. Compound 37, the known epimer of 1, as
well as its derivatives 32 and 33 were also expected to
improve our understanding of the structural prerequi-
sites necessary for optimal inhibition of the enzyme
under scrutiny.
When compared to the ‘simple’ modifications in 3–5 as
well as 7, this strong contribution to the binding process
by the 1-N-substituent resulted in a re-gain of activity of
four orders of magnitude. Clearly, this observation
makes the aglycon binding site a highly interesting and
worthwhile target to explore.
N
NHC12H25
OH
O
H
N
HO
(CH2)7C(CH2)4OH
HO
HO
HO
OH
OH
12
13
Ph
N
N
Me
NH(CH2)5CH3
OH
HO
HO
HO
OH
OH
OH
14
15
Figure 3. Structures of compounds 12–15.

T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
3487
H
N
H
N
HO
NHCO
HO
NHSO2
HO
OH
16
HO
OH
17
NMe2
O
NEt2
O
H
N
H
N
HO
NHSO2
HO
NHCO
HO
OH
HO
OH
18
19
Figure 4. Structures of compounds 16–19.
O
R
HOOC
H
N
H
N
R
HO
NH2
HO
N
H
R'
R'
HO
OH
HO
OH
HBTU, Et3N
5
20: R = R’ = H
21: R = NMe2, R’ = H
22: R = H, R’ = NMe2
Scheme 1. Synthesis of compounds 20–22.
Compounds 20–22 were accessible by the same reaction
sequence, starting from known 1-amino-1,2,5-trideoxy-
To investigate the contributions of the diethylamino
group and the aromatic system of the strong inhibitor
19, the corresponding derivative 25 was synthesised
(Fig. 5) by treatment of compound 5 with coumarin-3-
carboxylic acid in the presence of HBTU (63% yield).
2,5-imino-D
-mannitol (5).4 The different phenyl substit-
uents were introduced by coupling of the corresponding
benzoic acid derivatives employing O-benzotriazol-
1-yl-N,N,N0,N0-tetramethyluronium hexafluoroborate
(HBTU) as the coupling reagent (Scheme 1).
Inhibitor 26 containing a vinylogous amide was pre-
pared from 5 in practically quantitative yield by reaction
with 1,3-dimethyl-5-[(dimethylamino)methylene]-2,4,6-
(1H, 3H, 5H)-trioxopyrimidine (DTPM reagent)13 in
methanol in the presence of Et3N (Fig. 5).
Yields ranged from 75% for the simple benzamide com-
pound 20 to 14% for the 3-dimethylaminobenzoyl
derivative 21 and 16% for the 4-substituted compound 22.
To address the influence of the carboxamide versus
sulfonamide linkage on biological activities, sulfona-
mides 23 and 24 were prepared by reaction of compound
5 with the respective sulfonyl chloride (Scheme 2).
For the synthesis of 2,5-dideoxy-6-O-hexyl-2,5-imino-
mannitol 27, 5-azido-5-deoxy-1,2-O-isopropylidene-3-
O-methoxymethyl-a-
-glucofuranose5 28 was employed
D-
D
as the starting material (Scheme 3). Compound 28 was
treated with sodium hydride and 1-bromohexane in
THF to obtain 29, followed by removal of the 1,2-O-
isopropylidene group with ion exchange resin Amberlite
IR 120 [Hþ] to yield free glucofuranose derivative 30. Its
conversion to the corresponding open chain 5-azido-6-
O-hexyl fructose derivative 31 was achieved exploiting
glucose isomerase (EC 5.3.1.5).14 The equilibrium of the
isomerisation reaction was found to be on the aldose
side. Consequently, remaining starting material had to
be oxidised with Br2 and BaCO3 to the corresponding
H
N
HO
NHSO2R
23: R = phenyl
24: R = octyl
5
HO
OH
Scheme 2. Synthesis of compounds 23 and 24.

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T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
H
H
O
N
N
H
N
HO
H
N
HO
NHC
O
O
O
O
HO
OH
HO
OH
N
Me
Me
O
25
26
Figure 5. Structures of compounds 25 and 26.
N3
N3
O
O
HO
O
CH3(CH2)5O
O
1-Br(CH2)5CH3
NaH
MOMO
O
MOMO
O
28
29
N3
O
CH3(CH2)5O
IR 120 H +
OH
EC 5.3.1.5
MOMO
OH
30
H
N
N3
OH
HO
O(CH2)5CH3
CH3(CH2)5O
Pd(OH) 2, H2
OH
OH
O
HO
OH
31
27
Scheme 3. Synthesis of compounds 27.
gluconolactone, which could be separated from 31 by
chromatography. Hydrogenation of 31 with concomi-
tant intramolecular reductive amination with Pd(OH)2/
39. Subsequent isomerisation with glucose isomerase to
the corresponding -sorbose derivative 40 followed by
oxidation of the remaining aldose allowed access to pure
40. Its hydrogenation gave the desired 1-O-hexyl-imi-
noalditol 32 (Scheme 4).
L
C gave the desired 2,5-dideoxy-6-O-hexyl-2,5-imino-
mannitol 27.
D-
To investigate the influence of the stereochemistry at
position C-2, two 2,5-imino-2,5-dideoxy- -glucitol
derivatives 32 and 33 were synthesised. Starting from 5-
azido-5-deoxy-1,2-O-isopropylidene-
-idofuranose15 34
protection of position O-6 was achieved as the t-butyl-
dimethylsilyl ether 35. Protection of the remaining free
hydroxyl group at position 3 with chloromethyl methyl
ether gave 36 and deprotection of the silyl ether
employing tetrabutylammonium fluoride furnished
The 6-O-phenylpropyl derivative 33 was synthesised
accordingly. Following the above sequence employing 1-
D
bromophenylpropane in the reaction of
(37) to derivative 41, acidic deprotection (42), enzymatic
isomerisation to the -sorbose derivative 43 and sub-
sequent intramolecular reductive amination gave 2,5-
L-idofuranose
L
L
dideoxy-2,5-imino-6-O-(1-phenylpropyl)-D-glucitol 33
in fair overall yields.
L
-idofuranose derivative 37. Reaction of the liberated
hydroxyl group with bromohexane in the presence of
sodium hydride gave 38. Treatment with acidic ion
exchange resin Amberlite IR 120 [Hþ] in acetonitrile–
3. Biological activities
Ki values were measured with b-glucosidase from
Agrobacterium sp. in the presence of the corresponding
water furnished 5-azido-5-deoxy-6-O-hexyl-L-idofuranose

T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
3489
N3
N3
O
O
HO
O
TBDMSO
O
TBDMSCl
imidazole
MOMCl
HO
O
HO
O
34
35
N3
N3
O
O
TBDMSO
O
HO
O
TBAF
RBr, NaH
MOMO
O
MOMO
O
36
37
N3
N3
O
O
RO
O
RO
E C 5.3.1.5
IR 120 H +
OH
MOMO
O
MOMO
OH
38: R = (CH2)5CH3
41: R = (CH2)3Ph
39: R = (CH2)5CH3
42: R = (CH2)3Ph
H
N
N3
OH
HO
OR
CH3(CH2)5O
Pd(OH) 2, H2
OH
OH
O
HO
OH
40: R = (CH2)5CH3
43: R = (CH2)3Ph
32: R = (CH2)5CH3
33: R = (CH2)3Ph
Scheme 4. Synthesis of compounds 32 and 33.
inhibitors using para-nitrophenyl-b-glucoside as sub-
strate ([S]ꢀ1.5 Km) in 50 mM NaPi buffer containing
0.1% BSA at pH 7.
comparing two bicyclic aromatic substituents, the un-
substituted napthoyl (16) and coumarin carboxamide
(25) compounds, the heterocyclic coumarin system of-
fers an advantage of one order of magnitude in
inhibitory power.
From the data obtained for the compounds in Table 1,
the following conclusions can be drawn: All compounds
screened are competitive inhibitors of the enzyme under
scrutiny. Amides as well as sulfonamides at the 1-posi-
tion (9, 10, 24) are generally better inhibitors than the
corresponding 1-ethers (8, 27). There are no significant
differences in activity between structurally equivalent
amides and sulfonamides such as between 20 and 23, 10
and 24 as well as 16 and 17.
Bicyclic aromatic systems featuring tertiary amine sub-
stituents in the distal ring such as 18 and 19 have been
found most powerful, thus far, with improvements of
two orders of magnitude when compared to parent
compound 1 as well as similarly strong improvements
over their corresponding unsubstituted relatives (17 and
25, respectively). They compare favourably with the
most active compounds known to date.
Medium length, simple unsubstituted aliphatic substit-
uents (e.g., 10 and 27) are distinctly better than short
chains (4, 7). They are slightly better than terminally
functionalised relatives (8 and 9) with no additional
effect of increased chain length (12), and are compa-
rable to the simple unsubstituted (20) and amine
substituted monocyclic aromatic systems (21 and 22) as
well as the unsubstituted napthyl derivatives (16, 17)
probed in this study. Small improvements were de-
tected with amine substituted benzamides (21 and 22)
over the unsubstituted compound (20). Interestingly,
The distance of the amine substituted aromatic system
from the inhibitor moiety was not found crucial in the
one example (18 vs 34) probed.
Compound 26 is half as active as the parent compound
1, but still in the sub-micromolar range. This type of
substituent may provide a new entry to a group of
compounds in which additional hydrogen bonding be-
tween the inhibitor and the enzyme may strengthen the
sugar–protein interaction.

Table 1. Activities (Ki, lM, pH ¼ 7.0) of C-1 derivatives of DMDP against Agrobacterium sp. b-glucosidase
H
N
H
N
H
N
H
N
H
N
H
N
HO
HO
OH
F
HO
OMe
HO
NH2
HO
NH(CH2)6OH
HO
HO
OH
HO
OH
HO
OH
HO
OH
HO
OH
HO
OH
OH
55 Ki = 25
13 Ki = 0.2
43 = 10
212 Ki = 12
33 Ki = 30
Ki
65 Ki = 10
H
N
H
N
H
N
H
N
H
N
O
O
O
O
HO
NHCCH3
HO
O(CH2)6OH
NHC(CH2)5OH
HO
NHC(CH2)4CH3
HO
NHC(CH2)2Ph
HO
H
N
HO
NHC
HO
OH
HO
OH
O
HO
OH
HO
OH
HO
OH
HO
OH
105 Ki = 0.1
75 Ki = 25
85 Ki = 1.2
95 Ki = 0.3
118 Ki = 0.15
168 Ki = 0.55
NMe2
NMe2
Me 2
N
O
O
NEt2
H
N
H
H
H
N
HO
NHSO2
H
N
H
N
N
N
HO
NHC
O
HO
NHSO2
HO
NHC
HO
NHC
HO
NHC
O
HO
OH
O
O
HO
OH
HO
OH
HO
OH
HO
OH
HO
OH
198 Ki = 0.0012
22 = 0.27
1812 Ki = 0.0024
178 Ki = 0.10
Ki
21 Ki = 0.39
20 Ki = 0.88
H
N
H
HO
N
H
O
O
H
N
H
N
H
N
H
N
H
O
O
HO
NHSO2
N
HO
O(CH2)5CH3
HO
O(CH2)5CH3
HO
NHSO2(CH2)7CH3
HO
OH
HO
NHC
N
N
O
Me
HO
OH
Me
HO
OH
HO
OH
HO
OH
HO
OH
O
24 Ki = 0.2
23 Ki = 1.0
25 Ki = 0.037
26 Ki = 0.44
27 = 0.74
Ki
32 Ki = 120
H
H
N
H
N
H
N
O
NHC(CH2)10CH3
H
N
N
SO2NHCH2CH2OH
HO
OH
HO
NH(CH2)3NH2
HO
O(CH2)3Ph
HO
HO
NHC(CH2)5NHSO2
O
HO
OH
HO
OH
HO
OH
HO
OH
HO
OH
NMe2
NMe2
3512 Ki = 420
368 Ki = 0.1
3412 Ki = 0.0021
33 Ki = 50
Ki
37 = 19
385 Ki
= 15

T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
3491
2.5
2
19
34
18
For inhibitors stronger than 1:
Multiples of activities of compounds x compared to parent
compound 1, as expressed by log [(Ki (1)/Ki (x)].
1.5
1
25
0.5
0
10 12 17
11
1
22
23
9
21
26
-0.5
-1
16
27
20
24
8
For inhibitors weaker than 1:
Fractions of activities of compounds x compared to parent
compound 1, as expressed by -log [(Ki (x)/Ki (1)].
-1.5
-2
4
6
38
7
3
5
-2.5
Figure 6. Comparison of Ki values of compounds as multiples, or fractions, respectively, of Ki (1).
Two gluco-configured derivatives, 32 and 33, epimeric to
compound 1 at position C-2 were synthesised, carrying a
hexyl and a propylphenyl ether substituent, respectively,
at C-1. Not unexpectedly, compared to their C-2 epi-
mers, both compounds showed only weak interaction
with b-glucosidase from Agrobacterium sp (Fig. 6).
itly. Mass spectroscopy was conducted on a HP 1100
series MSD, Hewlett Packard. The scan mode for po-
sitive ions (mass range 100–1000 D) was employed
varying the fragmentation voltage from 50 to 250 V with
best molecular peaks observed at 150 V.
Agrobacterium sp. b-glucosidase was purified and as-
sayed as described.16 Kinetic studies were performed
at 37 ꢁC in pH 7.0 sodium phosphate buffer (50 mM)
containing 0.1
% bovine serum albumin, using
4. Conclusion
7.2 · 10ꢁ5 mg/mL enzyme. Approximate values of Ki
were determined using a fixed concentration of sub-
We have synthesised a range of new derivatives of the
strate, 4-nitrophenyl b-
D
-glucopyranoside (0.11 mM ¼
powerful glucosidase inhibitor 2,5-dideoxy-2,5-imino-
mannitol as well as selected relatives of the corre-
sponding -gluco epimer with different substituents at
D-
1:5 ꢂ Km) and inhibitor concentrations ranging from 0.2
times to 5 times the Ki value ultimately determined.
A horizontal line drawn through 1=Vmax in a Dixon
plot of this data (1=v vs [S]) intersects the experimental
line at an inhibitor concentration equal to Ki. Full Ki
determinations were performed using the same range of
inhibitor concentrations while also varying substrate (4-
nitrophenyl glucoside) concentrations from approxi-
mately 0.015–0.6 mM. Data were analysed by direct fit
D
positions C-1 and C-6, respectively. These compounds
were found to be competitive inhibitors of the Agro-
bacterium sp. b-glucosidase employed in this study.
Clearly, it could be shown that various lipophilic sub-
stituents generally increased the activity and that an
additional alkylamino group attached to aromatic sys-
tems further improved the interaction between the imi-
nosugar and the enzyme. The size of the aromatic
system was also found to be very important for strong
inhibitory activity.
to the Michaelis Menten equation describing reaction in
17
the presence of inhibitors using the program GRAFIT.
5.1. 1-Benzoylamino-1,2,5-trideoxy-2,5-imino-
(20)
D-mannitol
5. Experimental
Compound 54 (98 mg, 0.6 mmol) was dissolved in 10 mL
methanol, to the cooled (0 ꢁC) solution HBTU (230 mg,
0.6 mmol), Et3N (0.1 mL, 0.71 mmol) and benzoic acid
(41 mg, 0.3 mmol) were added and the reaction was
stirred at room temperature for 1 h. During this period
3 mL DMF were slowly added for solubility reasons.
The solvents were removed under reduced pressure and
the crude product was purified by silica gel chroma-
Thin layer chromatography (TLC) was performed on
precoated aluminium plates (Merck 5554) employing 5%
vanillin/sulfuric acid as well as ceric ammonium
molybdate as staining agents. For column chromato-
graphy, silica gel 60, 230–400 mesh (Merck 9385), was
1
used. H NMR spectra were recorded on a Varian IN-
OVA 500 operating at 499.925 MHz. 13C NMR spectra
were recorded at 75.47 or 125 MHz. Residual nondeu-
terated solvent was used as internal standard for deter-
mination of chemical shifts. Signals of aromatic as well
as alkyl substituents including protecting groups were
found in the expected regions and are not listed explic-
tography using CHCl3/MeOH (v/v 5/1 containing 1%
20
D
NH4OH) to give 120 mg (74.5%). ½aꢃ +22.0 (c 0.5,
H2O). 13C NMR (D2O): d 1712 (C@O), 79.3, 77.6 (C-3,
C-4), 61.7, 61.6, 59.9 (C-2, C-5, C-6), 42.2 (C-1). H
1
NMR (D2O): d 3,80–3.73 (2d, 2H, J2;3¼4;5 3.8, H-3, H-4),

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T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
3.61 (dd, 1H, J6;6 ¼ 11:8, J6 ;5 ¼ 4:4, H-60), 3.53 (dd, 1H,
J5;6 ¼ 6:2, H-6), 3.49–3.39 (m, 2H, H-2, H-5), 3.13 (dd,
J3;4 ¼ 6:3, J2;3 ¼ 7:3, H-3), 3.82 (dd, 1H, J4;5 ¼ 7:8, H-4),
0
0
0
3.76 (dd, 1H, J6;6 ¼ 12:6, J5;6 ¼ 3:9, H-6), 3.68 (dd, 1H,
J5;6 ¼ 5:9, H-60), 3.41–3.36 (m, 2H, H-2, H-5) 3.13 (d,
0
0
1H, J1;1 ¼ 12:2, J1;2 ¼ 5:9, H-1), 2.98 (dd, 1H,
J1 ;2 ¼ 4:8, H-10).
2H, J1 ;2 ¼ 5:9, H-1, H-10).
0
0
5.2. 1-(3-Dimethylamino)benzoylamino-1,2,5-trideoxy-
2,5-imino-
5.5. 1-(n-Octylsulfonyl)amino-1,2,5-trideoxy-2,5-imino-
D-mannitol (24)
D
-mannitol (21)
Compound 5 (90 mg, 0.56 mmol) and 3-dimethylami-
nobenzoic acid (91.6 mg, 0.55 mmol), HBTU (209 mg,
0.55 mmol) and Et3N (0.1 mL, 0.71 mmol) were reacted
To a 5% solution of 1-amino-1,2,5-trideoxy-2,5-imino-
D-mannitol 5 (39 mg, 0.24 mmol) in DMF (1.2 mL) octyl
sulfonic chloride (50 lL, 0.24 mmol) and Et3N (three
drops) were added and the reaction stirred at room
temperature for 3 h. TLC (CHCl3/MeOH/NH4OH
1:2:1) indicated completed reaction. The mixture was
concentrated under reduced pressure and purified by
silica gel chromatography (CH2Cl2/MeOH/NH4OH
20
to obtain product 21 (24 mg, 14%): ½aꢃ +24.4 (c 0.6,
D
H2O). 13C NMR (CD3OD): d 171.3 (C@O), 77.9, 76.4
(C-3, C-4), 64.6 (C-5), 63.4 (C-2), 59.7 (C-6), 40.8 (C-1),
39.6 (2 C, 2 · NCH3). 1H NMR: 3.82 (m, 2H, J3;4 ¼ 5:8,
0
H-3, H-4), 3.71 (dd, 1H, J5;6 ¼ 3:9, J6;6 ¼ 11:2, H-6),
3.63 (dd, 1H, J5;6 ¼ 2:4, H -60), 3.61 (m, 1H, J1 ;2 ¼ 5:4,
500:100:1). Product 23 was obtained as white semicrys-
0
0
20
H-10), 3.54 (dd, 1H, J1;2 ¼ 7:3, J1;1 ¼ 13:7, H-1), 3.24 (m,
talline solid (30.2 mg, 42%): ½aꢃ +23.0 (c 0.2, MeOH).
0
D
1H, H-2), 3.08 (m, 1H, H-5), 2.99 (2s, 6H, 2CH3). MS
([MHþ] m=z) Calcd for C15H23O4N3: 309.41; Found:
[M+H]þ 310.35.
13C NMR (CD3OD): d 78.0, 76,9 (C-3, C-4), 64.5, 62.8,
60.4 (C-2, C-5, C-6), 51.5 (C-7), 43.3 (C-1), 31.8 (C-8),
29.1, 29.0, 28.2, 23.5, 22,5 (C-9, C-10, C-11, C-12, C-13),
13.2 (C-14). 1H NMR: d 3.91 (dd, 1H, J2;3 ¼ 5:9,
J3;4 ¼ 5:40, H-3), 3.87 (dd, 1H, J3;4 ¼ J4;5 ¼ 5:4, H-4),
0
5.3. 1-(4-Dimethylamino)-benzoylamino-1,2,5-trideoxy-
-mannitol (22)
3.79 (dd, 1H, J6;6 ¼ 11:2, J6;5 ¼ 3:9, H-6), 3.70 (dd, 1H,
J6 ;5 ¼ 6:3, H-60), 3.40–3.26 (m, 4H, H-2, H-5, H-1, H-
0
2,5-imino-
D
10), 3.11 (t, 2H, J ¼ 7.8, H-7), 1.78 (m, 2H, H-8), 1.45
(m, 2H, H-9), 1.40–1.28 (m, 8H, H-10, H-11, H-12, H-
13), 0.92 (t, 3H, J ¼ 6.8, H-14).
A 5% solution of 1-amino-1,2,5-trideoxy-2,5-imino-
D
-
mannitol 5 (53 mg, 0.327 mmol) was dissolved in DMF
(5 mL) and the mixture was cooled to 0 ꢁC. 4-Dimeth-
ylaminobenzoic acid (55 mg, 0.33 mmol), HBTU
(131 mg, 0.35 mmol) and Et3N (71 lL, 0.50 mmol) were
added and the mixture was stirred at 25 ꢁC. After 4.5 h,
TLC indicated completed reaction. The mixture was
concentrated under reduced pressure and purified by
silica gel chromatography (CHCl3/MeOH/NH4OH
5.6. 1-Amino-N-(coumarin-3-oyl)-1,2,5-trideoxy-2,5-imino-
D
-mannitol (25)
A 10% solution of 1-amino-1,2,5-trideoxy-
D
-mannitol 5
(72 mg, 0.44 mmol) in DMF was cooled to 0 ꢁC and
consecutively coumarin-3-carboxylic acid (84 mg,
0.44 mmol), HBTU (167,4 mg, 0.45 mmol) and Et3N
(0.1 mL, 0.71 mmol) were added and the reaction was
stirred at room temperature for 4 h. The mixture was
concentrated under reduced pressure and chromato-
graphed on silica gel (CHCl3/MeOH/NH4OH
100:500:6). Product 22 was obtained as white semicrys-
20
D
talline solid (16 mg, 16%): ½aꢃ +22.5 (c 0.8, H2O). 13C
NMR (D2O): d 170.3 (C@O), 75.8, 74.3 (C-3, C-4), 62.9
(C-5), 61.4 (C-6), 58.1 (C-2), 46.4 (2C, CH3), 39.4 (C-1).
1H NMR: d 4.08 (m, 2H, H-3, H-4), 3.90 (dd, 1H,
J5;6 ¼ 3:7, J6;6 ¼ 12:7, H-6); 3.83 (m, 3H, H-1, H-10, H-
0
60), 3.72 (m, 1H, H-5), 3.62 (m, 1H, H-2), 3.21 (2s, 6H,
2CH3). MS ([MHþ] m=z) Calcd for C15H24O4N3: 310.41;
Found: [M+H]þ 310.34.
100:600:7). Product 25 was obtained as white glass
20
D
(92.5 mg, 63%): ½aꢃ +20.7 (c 0.8, H2O). 13C NMR
(CD3OD): d 79.9, 77.9 (C-3, C-4), 63.7, 61.6, 61.3 (C-2,
C-5, C-6), 42.4 (C-1). 1H NMR: d 3.80 (2 dd, 2H,
J6;6 ¼ 13:2, J5;6 ¼ J5;6 ¼ 6:7, H-6, H-60), 3.70 (dd, 1H,
0
0
0
5.4. 1-(Benzenesulfonyl)amino-1,2,5-trideoxy-2,5-imino-
D
J4;5 ¼ 3:9, J3;4 ¼ 11:2, H-4), 3.67 (dd, 1H, J1;1 ¼ 13:2,
J1 ;2 ¼ 5:4, H-10), 3.62 (dd, 1H, J2;3 ¼ 5:9, H-3), 3.56 (dd,
0
-mannitol (23)
1H, J1;2 ¼ 6:8, H-1), 3.21 (m, 1H, H-2), 3.03 (m, 1H, H-
5). MS([MHþ] m=z) Calcd for C16H18O6N2: 334.4;
Found: [M+H]þ 335.1.
To a 5% solution of 1-amino-1,2,5-trideoxy-2,5-imino-
-mannitol 5 (39 mg, 0.24 mmol) in MeOH (1.2 mL),
D
benzenesulfonic chloride (31.0 lL, 0.24 mmol) was ad-
ded and the pH 7–8 was adjusted by addition of Et3N
(24.7 lL, 0.18 mmol). The reaction mixture was stirred
at room temperature for 2 h, when TLC (CHCl3/MeOH/
NH4OH 1:2:1) indicated completed conversion. The
mixture was concentrated under reduced pressure and
purified by silica gel chromatography (CH2Cl2/MeOH/
5.7. 1-[N-(1,3-Dimethyl-2,4,6 (1H, 3H, 5H)-trioxopyr-
imidine-5-ylidene)methyl]amino-1,2,5-trideoxy-2,5-imino-
D
-mannitol (26)
The starting material 5 (57.2 mg, 0.32 mmol) was dis-
solved in 2 mL methanol, Et3N (88 lL, 0.36 mmol) and
1,3-dimethyl-5-[(dimethylamino)methylene] 2,4,6 (1H,
3H, 5H)-trioxopyrimidine (DTPM reagent)13 (75 mg,
0.36 mmol) dissolved in 200 lL MeOH were added in
NH4OH 400:100:1). Product 23 was obtained as white
20
D
glass (30.2 mg, 42%): ½aꢃ +23.9 (c 0.6, MeOH). 13C
NMR (D2O): d 76.1, 75.0 (C-3, C-4), 62.9, 61.0, 58.8 (C-
2, C-5, C-6), 41.8 (C-1). 1H NMR: d 3.90 (dd, 1H,

T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
3493
one portion and the reaction was stirred at room tem-
5.10. 5-Azido-5-deoxy-6-O-hexyl-D-fructose (31)
perature for 1 h to give 26 (108.6 mg, 99 %) as a white
precipitate.13C NMR (D2O): d 179.5 (C@O), 164.5,
162.5, 160.8, 153.0 (DTPM), 90.5 (DTPM), 78.6, 76.8
(C-3, C-4), 61.8, 61.0, 59.7 (C-2, C-5, C-6), 52.6 (C-1).1H
NMR (D2O): d 3.80 (m, 2H, J2;3 ¼ J3;4 ¼ 3:8, H-3, H-4),
A 5% aqueous solution of the free aldose 30 was spun on
the rotary evaporator in the presence of immobilised
glucose isomerase (Sweetzyme T, EC 5.3.1.5, 1.5 g) at
60 ꢁC for 12 h. The solids were filtered off, BaCO3
(643 mg, 3.3 mmol) and bromine (150 mg, 0.94 mmol)
were added and the solution was stirred at ambient
temperature until the remaining aldose was converted
3.68 (dd, 1H, J6;6 ¼ 10:8, J5;6 ¼ 4:7, H-60), 3.60 (dd, 1H,
J5;6 ¼ 5:5, H-6), 3.58–3.52 (m, 2H, H-2, H-5), 3.24–3.22
(m, 1H, H-10), 3.06 (s, 6H, 2CH3), 2.99–296 (m, 1H,
H-1).
0
0
into the less polar
D-gluconotactone. Air was bubbled
through the solution to remove the excess of bromine.
After filtration, the solution was concentrated under
reduced pressure and the crude product was purified by
silica gel chromatography (cyclohexane/ethyl acetate 4/1
5.8. 5-Azido-5-deoxy-6-O-hexyl-1,2-O-isopropylidene-3-
O-methoxymethyl-a-D-glucofuranose (29)
v/v) to give pure ketose 31 (30 mg, 8%): ½aꢃ20 )40.8 (c 0.4,
D
To a 5% solution of 285 (630 mg, 2.18 mmol) in THF
containing 20% DMF, sodium hydride (55% dispersion
in oil, 638.2 mg, 15.2 mmol) and 1-bromohexane
(770.0 lL, 5.5 mmol) were added. The mixture was stir-
red at room temperature until the starting material was
no longer detectable by TLC (4 h). The heterogeneous
mixture was quenched by addition of excess MeOH.
Solvents were evaporated under reduced pressure, the
remaining residue was diluted with CH2Cl2, washed
with 5% aqueous HCl, satd aqueous NaHCO3 and dried
over Na2SO4. The filtrate was concentrated under re-
duced pressure and the residue was purified by silica gel
CH2Cl2); 13C NMR (CDCl3): d 212.4 (C-2), 75.6 (C-3),
72.7 (C-4), 72.4 (C-7), 71.0 (C-6), 66.7 (C-1), 60.8 (C-5),
1
31.8, 29.7, 25.9, 21.3, 14.2. H NMR: d 4.60 (d, 1H,
J ¼ 19:8, H-1), 4.49–4.41 (m, 2H, H-10, H-3), 4.00 (m,
1H, H-4), 3.72–3.56 (m, 3H, H-5, H-6, H-60), 3.52 (m,
2H), 1.52 (m, 2H), 1.35 (m, 6H), 0.89 (t, 3H).
5.11. 2,5,6-Trideoxy-1-O-hexyl-2,5-imino-D-mannitol
(27)
Hydrogenation and concomitant intramolecular reduc-
tive amination of ketose 31 (30 mg, 0.1 mmol) was con-
ducted in dry methanol in the presence of a catalytic
amount of Pd(OH)2/C (20%) under hydrogen atmo-
sphere at ambient pressure. After 2 h, TLC (CHCl3/
MeOH/NH4OH 2:2:1) showed no further starting
material. The catalyst was removed by filtration, the
solution was concentrated and the resulting yellow syrup
was purified by silica gel chromatography(CCl3/MeOH/
chromatography (cyclohexane/ethyl acetate 10:1 v/v) to
20
D
yield pure compound 29 (595 g, 76%): ½aꢃ )17.0 (c 1.0,
CH2Cl2); 13C NMR (CDCl3): d 112.3, 105.4 (C-1), 96.9,
82.9 (C-2), 80.3 (C-3), 78.5 (C-4), 71.8, 71.7 (C-6), 58.9
1
(C-5), 56.3, 31.9, 29.8, 27.0, 26.5, 25.9, 22.8, 14.2. H
NMR: d 5.86 (d, 1H, J1;2 ¼ 3:5, H-1), 4.74 (dd, 2H,
J ¼ 6:6), 4.6 (d, 1H, H-2), 4.15 (m, 1H, J3;4 ¼ 2:6, H-3),
4.03 (dd, 1H, J4;5 ¼ 9:7, H-4), 3.83 (m, 1H, J5;6 ¼ 2:2,
0
0
J5;6 ¼ 6:2, H-5), 3.59 (dd, 1H, J6;6 ¼ 10:1, H-6), 3.48
(dd, 1H, H-60), 3.44 (s, 2H), 1.56 (m, 2H), 1.46 (2s,
2 · 3H),1.29 (m, 6H), 0.87 (m, 3H).
NH4OH 120/10/1) to furnish inhibitor 27 (8 mg, 31%) as
20
D
a colourless oil: ½aꢃ +16.9 (c 0.1, MeOH); 13C NMR
(CD3OD): d 78.5, 78.4 (C-3, C-4), 71.4 (C-1), 70.9 (C-6),
63.5 (C-7), 61.8 (C-5), 61.8 (C-2), 34.2 (C-10), 31.7 (C-9),
1
25.8 (C-8), 22.5 (C-11), 13.2 (C-12). H NMR: d 3.79–
5.9. 5-Azido-5-deoxy-6-O-hexyl-a-
D
-glucofuranose (30)
3.78 (m, 2H, H-3, H-4), 3.69–3.42 (m, 2H, H-1, H-10, H-
2, H-6, H-60), 3.33 (m, 2H, H-7), 3.09 (m, 1H, H-5), 1.59
(m, 2H, H-8), 1.37 (m, 6H, H-9, H-10, H-11), 0.91 (t,
3H, H-12).
A 5% solution of intermediate 29 (1.0 g, 2.68 mmol) in
50% aqueous CH3CN was stirred with 15 mL of ion-
exchange resin Amberlite IR-120 [Hþ] at 45 ꢁC until
TLC showed quantitative conversion into a single, more
polar product. The resin was removed by filtration, the
filtrate was concentrated under reduced pressure and the
crude product was purified by silica gel chromatography
5.12. 5-Azido-6-O-t-butyldimethylsilyl-5-deoxy-1,2-O-
isopropylidene-b-L-idofuranose (35)
(ethyl acetate) to give free
D
-glucofuranose derivative 30
To a 20% solution of 5-azido-5-deoxy-1,2-O-isopropyl-
idene-
-idofuranose (34)15 (3.0 g, 12.2 mmol) in dry
20
(500 mg, 71%): ½aꢃ )27.5 (c 0.4, CH2Cl2); b-Anomer:
L
D
13C NMR (CDCl3): d 103.5 (C-1), 81.1 (C-4), 77.2 (C-2),
76.2 (C-3), 72.1 (C-7), 70.6 (C-6), 60.6 (C-5), 31.8, 29.7,
25.8, 22.8, 14.2. a-Anomer: 13C NMR (CDCl3): d 97.1
(C-1), 79.9 (C-4), 77.9 (C-2), 76.6 (C-3), 72.2, 70.9 (C-6),
DMF imidazole (2.1 g, 30.6 mmol) and t-butyl-dimeth-
ylsilyl chloride (2.2 g, 14.7 mmol) were added. The reac-
tion was stirred at 40 ꢁC until TLC (cyclohexane/ethyl
acetate 1/1) confirmed quantitative conversion (2 h). The
mixture was diluted with water, extracted with CH2Cl2
twice, the organic phase was dried and concentrated
under reduced pressure. Silica gel chromatography
(cyclohexane/ethyl acetate 5/1 v/v) gave silylated com-
1
60.1 (C-5), 31.8, 29.7, 25.8, 22.8, 14.2. H NMR: d 5.55
(d, 1H, J1;2 ¼ 2:4, H-1b), 5.20 (s, 1H, H-1a), 4.29 (s, 1H,
H-2a), 4.22–4.17 (m, 2H, J3;4 ¼ 3:4, J4;5 ¼ 8:8, H-3a, H-
3b, H-4a, H-4b), 4.11 (m, 1H, H-2b), 3.93 (m, 1H,
J5;6 ¼ 2:9, J5;6 ¼ 6:1, H-5a), 3.88–3.83 (m, 2H,
pound 35 (4.40 g, 100%) as a colourless syrup: ½aꢃ20 )11.5
0
D
J6;6 ¼ 10:3, H-6a, H-5b), 3.79 (dd, 1H, H-6b), 3.67 (dd,
(c 0.8, CH2Cl2); 13C NMR (CDCl3): d 112.0 (C-7), 104.8
(C-1), 85.2, 82.6 (C-2, C-4), 75.5 (C-3), 63.5, 62.2 (C-5, C-
6). 1H NMR: d 5.85 (d, 1H, J1;2 ¼ 3:5, H-1), 4.42 (d, 1H,
0
1H, H-60a), 3.62 (dd, 1H, H-60b), 3.51 (m, 4H), 1.60 (m,
4H), 1.32 (m, 12H), 0.89 (t, 6H).

3494
T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
H-2), 4.07 (s, 1H, H-3), 3.99 (dd, 1H, J4;5 ¼ 7:0, H-4),
J4;5 ¼ 8:8, H-4), 4.05 (d, 1H, J3;4 ¼ 3:1, H-3), 3.85 (m,
0 0
0
3.67 (m, 1H, J5;6 ¼ 5:7, J5;5 ¼ 9:2, H-5), 3.59 (m, 1H,
1H, J5;6 ¼ 3:5, J5;6 ¼ 6:2, H-5), 3.56 (m, 1H, J6;6 ¼ 11:6,
J6;6 ¼ 11:0, H-6), 3.54 (m, 1H, H-60).
H-6), 3.49 (m, 1H, H-60).
0
5.13. 5-Azido-6-O-t-butyldimethylsilyl-5-deoxy-1,2-O-
isopropylidene-3-methyloxymethyl-b-L-idofuranose (36)
5.16. 5-Azido-5-deoxy-6-O-hexyl-b-L-idofuranose (39)
Deprotection of compound 38 (780 mg, 2.09 mmol)
employing excess ion exchange resin Amberlite IR-120
A 5% solution of compound 35 (2.7 g, 7.5 mmol) in dry
DMF was treated with ethyl-N,N-diisopropylamine
(5.1 mL, 30 mmol) and freshly prepared MOM chlo-
ride18 (50% in MeOH, 3.6 mL, 22 mmol). The mixture
was stirred at room temperature until the starting
material was no longer detectable by TLC (30 h). Sol-
vents were evaporated under reduced pressure, the
remaining residue was diluted with CH2Cl2, consecu-
tively washed with 5% aqueous HCl, satd aqueous
NaHCO3 and dried over Na2SO4. The filtrate was
concentrated under reduced pressure and the residue
was purified by silica gel chromatography (cyclohexane/
[Hþ] in 50% aqueous acetonitrile gave compound 39
20
D
(404 mg, 67%): ½aꢃ + 35.1 (c 1.1 CH2Cl2); 13C NMR
(CDCl3): d b-Anomer: 103.2 (C-1), 84.6 (C-4), 80.0 (C-
2), 77.0 (C-3), 72.4, 70.4 (C-6), 62.0 (C-5), 31.7, 29.6,
25.8, 22.8, 14.2. a-Anomer: 96.6 (C-1), 81.2 (C-4), 77.9
(C-2), 76.2 (C-3), 72.4, 70.3 (C-6), 60.7 (C-5), 31.7, 29.6,
25.8, 22. 8, 14.2. 1H NMR: d 5.58 (m, 1H, H-1a), 5.22 (s,
1H, H-1b), 4-58–4.26 (m, 5H, H-2a, H-2b, H-3b, H-4a,
H-4b), 4.30 (d, 1H, H-3a), 3.92 (m, 1H, J5;6 ¼ 4:3,
0
J5;6 ¼ 8:3, H-5b), 3.82 (m, 1H, H-5a), 3.62 (dd, 2H,
J6;6 ¼ 10:8, H-6a, H-6b), 3.52–3.45 (m, 6H, H-60a, H-
0
ethyl acetate 6:1 v/v) to yield compound 36 (2.46 g,
60b, H-7), 1.58 (m, 4H), 1.30 (m, 12H), 0.88 (t, 6H).
20
D
81%): ½aꢃ )38.5 (c 0.8, CH2Cl2); 13C NMR (CDCl3): d
112.3 (C-7), 104.8 (C-1), 96.5 (C-14), 83.3, 80.8 (C-2, C-
4), 78.8 (C-3), 63.4, 62.3 (C-5, C-6). 1H NMR: d 5.85 (d,
1H, J1;2 ¼ 4:0, H-1), 4.63 (m, 2H, H-14), 4.52 (d, 1H, H-
2), 4.16 (dd, 1H, J4;5 ¼ 7:9, H-4), 3.97 (d, 1H, J3;4 ¼ 3:1,
5.17. 5-Azido-5-deoxy-6-O-hexyl-L-sorbose (40)
Employing immobilised glucose isomerase (Sweetzyme
T, EC 5.3.1.5, 1.5 g) to 5-azido-5-deoxy-6-O-hexyl-b-L-
0
H-3), 3.70 (m, 1H, J5;6 ¼ 5:2, J5;5 ¼ 8:0, H-5), 3.64 (m,
1H, H-6), 3.59 (m, 1H, H-60).
idofuranose39 (400 mg, 1.38 mmol) at 60 ꢁC gave prod-
20
uct 40 (94 mg, 24%): ½aꢃ +30.0 (c 0.2 CH2Cl2). 13C
D
NMR (CDCl3): d 211.2 (C-2), 75.6 (C-3), 73.1 (C-4),
72.5 (C-1), 67.3 (C-6), 61.6 (C-5), 31.8, 29.6, 25.9, 22.8,
5.14. 5-Azido-5-deoxy-1,2-O-isopropylidene-3-methyl-
-idofuranose (37)
1
oxymethyl-b-
L
14.2. H NMR: d 4.50 (d, 1H, J 17.6 Hz, H-1), 4.34 (d,
1H, H-10), 4.08 (m, 1H, H-3), 3.71 (m, 3H, H-4, H-6, H-
60), 3.45 (m, 3H, H-5, H-7), 1.53 (m, 2H), 1.20 (m, 6H),
0.83 (t, 3H).
To a 5% solution of compound 36 (2.7 g, 5.6 mmol) in
dry THF, tetrabutylammonium fluoride trihydrate
(2.7 g, 8.5 mmol) was added and the reaction was stirred
at 45 ꢁC until TLC (cyclohexane/ethyl acetate 1:1) indi-
cated completion (1 h). The solution was concentrated
under reduced pressure and the crude product was
purified by silica gel chromatography (cyclohexane/ethyl
5.18. 2,5-Dideoxy-6-O-hexyl-2,5-imino-D-glucitol (32)
The same procedure as for compound 27 applied to
5-azido-5-deoxy-6-O-hexyl- -sorbose 40 (75 mg, 0.26
mmol) employing Pd(OH)2/C under hydrogen atmos-
acetate 3:1 v/v) to give compound 37 in quantitative
L
20
D
yield (1.86 g): ½aꢃ )85.1 (c 1.3, CH2Cl2); 13C NMR
(CDCl3 ): d 112.3 (C-7), 104.8 (C-1), 96.6 (C-10), 83.3,
80.3 (C-2, C-4), 80.2 (C-3), 63.2, 62.2 (C-5, C-6). H
phere at ambient pressure gave product 32 (29 mg, 44%):
20
D
1
½aꢃ +12.5 (c 0.1, MeOH). 13C NMR (CD3OD): d 79.4
NMR: d 5.98 (d, 1H, J1;2 ¼ 4:0, H-1), 4.72 (m, 2H,
J ¼ 6:6, H-10), 4.63 (d, 1H, H-2), 4.25 (dd, 1H,
J4;5 ¼ 8:8, H-4), 4.08 (d, 1H, J3;4 ¼ 3:1, H-3), 3.87 (m,
(C-3), 77.9 (C-4), 71.4 (C-7), 69.5 (C-6), 66.3 (C-2), 62.3
(C-1), 60.1 (C-5), 31.7 (C-10), 29.6 (C-9), 25.8 (C-8), 22.5
1
(C-11), 13.2 (C-12). H NMR: 3.94 (dd, 1H, J3;4 ¼ 4:4,
0
0
1H, J5;6 ¼ 3:5, J5;5 ¼ 6:2, H-5), 3.73 (m, 1H, J6;6 ¼ 11:4,
H-3), 3.82 (dd, 1H, J4;5 ¼ 4:8, H-4), 3.67 (m, 2H, H-1,
H-10), 3.58–3.43 (m, 3H, H-5, H-6, H-60), 3.33 (m, 2H,
H-7), 2.97 (m, 1H, J5;6 ¼ 9:2, H-2), 1.57 (m, 2H, H-8),
1.32 (m, 6H, H-9, H-10, H-11), 0.91 (t, 3H, H-12).
H-6), 3.56 (m, 1H, H-60).
5.15. 5-Azido-5-deoxy-6-O-hexyl-1,2-O-isopropylidene-3-
methyloxymethyl-b-L-idofuranose (38)
5.19. 5-Azido-5-deoxy-6-O-(1-phenylpropyl)-1,2-O-iso-
propylidene-3-methyloxymethyl-b-L-idofuranose (41)
Applying the same procedure as for compound 29 to
compound 37 (750 mg, 2.60 mmol), employing 1-
bromohexane (900 lL, 6.5 mmol) and NaH (55% dis-
To a 5% solution of compound 37 (750 mg, 2.6 mmol) in
THF containing 20% DMF 1-bromophenylpropane
(1 mL, 6.6 mmol) and NaH (55% dispersion in oil,
persion in oil, 0.8 g, 19.0 mmol) product 38 (818 mg,
20
D
85%) was be obtained: ½aꢃ )40.9 (c 1.1, CH2Cl2); 13C
NMR: (CDCl3) d 112.3 (C-7), 104.8 (C-1), 96.7 (C-10),
83.3, 81.0 (C-2, C-4), 79.7 (C-3), 72.1 (C-12), 70.4 (C-6),
60.8 (C-5). 1H NMR: d 5.93 (d, 1H, J1;2 ¼ 3:5, H-1), 4.71
(dd, 2H, J ¼ 7:0, H-10), 4.61 (d, 1H, H-2), 4.22 (dd, 1H,
760 mg, 18.2 mmol) were added to obtain product 41 in
20
D
quantitative yield (1.06 g): ½aꢃ )36.9 (c 0.4, CH2Cl2).
13C NMR (CDCl3): d 142.0, 128.7, 128.6, 126.1 (phenyl),
112.4, 104.8 (C-1), 96.7 (MOM), 83.4, 80.9 (C-2, C-4),

T. M. Wrodnigg et al. / Bioorg. Med. Chem. 12 (2004) 3485–3495
3495
79.6 (C-3), 70.9, 70.5 (C-6), 60.8 (C-5), 56.5, 32.4, 31.4.
Acknowledgements
1H NMR: d 7.31–7.17 (m, 5H, phenyl), 5.95 (d, 1H,
J1;2 ¼ 3:5, H-1), 4.71 (dd, 2H, J ¼ 7:0), 4.61 (d, 1H, H-
2), 4.26 (dd, 1H, J4;5 ¼ 8:8, H-4), 4.06 (d, 1H, J3;4 ¼ 3:1,
MS measurements were conducted by Dr. M. Murkovic.
Glycogroup appreciate financial support by the Aus-
trian Fonds zur F€orderung der Wissenschaftlichen Fors-
chung (FWF), Vienna (Hertha-Firnberg-Fellowship
0
H-3), 3.88 (m, 1H, J5;6 ¼ 3:5, J5;6 ¼ 6:2, H-5), 3.59 (m,
1H, J6;6 ¼ 10:5, H-6), 3.44 (m, 2H), 3.49 (m, 1H, H-60),
0
€
3.40 (s, 3H), 2.70 (m, 2H), 1.91 (m, 2H).
T-18 CHE and Erwin Schrodinger-Stipend J2092 to
T.M.W. as well as Project 15726-N03). Financial sup-
port from the Protein Engineering Network of Centres
of Excellence of Canada (PENCE) is also gratefully
acknowledged.
5.20. 5-Azido-5-deoxy-6-O-(1-phenylpropyl)-L-idofura-
nose (42)
Deprotection of compound 41 (100 mg, 0.25 mmol)
employing ion exchange resin Amberlite IR-120 [Hþ]
20
D
gave product 42 (45 mg, 57%): ½aꢃ )17.84 (c 1.1,
References and notes
CH2Cl2). 13C NMR (CDCl3): d b-Anomer: 141.7, 128.7,
128.6, 126.3 (phenyl), 103.2 (C-1), 84.4 (C-4), 80.1 (C-2),
77.1 (C-3), 71.4 (C-7), 70.4 (C-6), 62.0 (C-5), 32.4 (C-8),
31.2 (C-9). a-Anomer: 141.7, 128.7, 128.6, 126.3 (phen-
yl), 96.6 (C-1), 80.9 (C-4), 77.9 (C-2), 76.2 (C-3), 71.5 (C-
7), 70.5 (C-6), 60.6 (C-5), 32.4 (C-8), 31.2 (C-9). 1H
NMR: d 7.19–7.06 (m, 10H, phenyl), 5.60 (d, 1H,
J1;2 ¼ 3:52, H-1a), 5.22 (s, 1H, H-1b), 4-58–4.26 (m, 5H,
H-2a, H-2b, H-3b, H-4a, H-4b), 4.30 (d, 1H, H-3a),
3.92–3.62 (m, 6H, H-5b, H-5a, H-6a, H-6bH-60a, H-
60b), 3.52–3.45 (m, 4H, H-7, H-9), 1.98 (m, 4H, H-8).
1. Welter, A.; Jadot, J.; Dardenne, G.; Marlier, M.; Casimir,
J. Phytochemistry 1976, 15, 747.
2. Iminosugars as Glycosidase Inhibitors; Stutz, A. E., Ed.;
VCH-WILEY: Weinheim, 1999 and references cited
therein.
3. Andersen, S. M.; Ebner, M.; Ekhart, C.; Gradnig, G.;
€
€
Legler, G.; Stutz, A. E.; Withers, S. G.; Wrodnigg, T. M.
Carbohydr. Res. 1997, 301, 155.
€
4. Wrodnigg, T. M.; Stutz, A. E.; Withers, S. G. Tetrahedron
Lett. 1997, 38, 5463.
€
5. Wrodnigg, T. M.; Gaderbauer, W.; Greimel, P.; Hausler,
H.; Sprenger, F. K. (Fitz); Stutz, A. E.; Virgona, C.;
€
Withers, S. G. J. Carbohydr. Chem. 2000, 19, 975.
6. Takaoka, Y.; Kajimoto, T.; Wong, C.-H. J. Org. Chem.
1993, 58, 4809.
7. Shibano, M.; Kitagawa, S.; Kusano, G. Chem. Pharm.
Bull. 1997, 45, 505.
5.21. 5-Azido-5-deoxy-6-O-(1-phenylpropyl)-L-sorbose
(43)
Isomerisation of compound 42 (600 mg, 1.86 mmol) with
immobilised glucose isomerase (Sweetzyme T, EC
€
8. Wrodnigg, T. M.; Withers, S. G.; Stutz, A. E. Bioorg.
Med. Chem. Lett. 2001, 11, 1063.
9. (a) Legler, G.; Finken, M.-T. Carbohydr. Res. 1996, 292,
103; (b) Legler, G.; Finken, M.-T.; Felsh, S. Carbohydr.
Res. 1996, 292, 91.
10. Panday, N.; Canac, Y.; Vasella, A. Helv. Chim. Acta 2000,
83, 58.
11. Ogawa, S.; Fujieda, S.; Sakata, Y.; Ishizaki, M.; Hisama-
tsu, S.; Okazaki, K. Bioorg. Med. Chem. Lett. 2003, 13,
3461.
5.3.1.5, 1.5 g) at 60 ꢁC gave product 43 (106 mg, 18%):
20
½aꢃ +46.0 (c 0.1, CH2Cl2). 13C NMR (CDCl3): d 210.8
D
(C-2), 141.6, 128.7, 128.7, 126.2 (phenyl), 75.7 (C-3), 72.9
(C-4), 71.4 (C-6), 71.2 (C-7), 67.3 (C-1), 61.7 (C-5), 32.4
(C-9), 31.2 (C-8). 1H NMR: d 4.97–4.19 (m, 5H, phenyl),
4.55 (d, 1H, J ¼ 18:0, H-1), 4.39 (d, 1H, H-10), 4.15 (m,
1H, H-3), 3.60 (m, 3H, H-4, H-6, H-60), 3.53 (m, 3H, H-5,
H-7, H-70), 2.70 (m, 2H, H-9), 1.94 (m, 2H, H-8).
€
12. Hermetter, A.; Scholze, H.; Stutz, A. E.; Withers, S. G.;
Wrodnigg, T. M. Bioorg. Med. Chem. Lett. 2001, 11,
1339.
13. Dekany, G.; Bornaghi, L.; Papageorgiou, J.; Taylor, S.
Tetrahedron Lett. 2001, 42, 3129.
5.22. 2,5-Dideoxy-6-O-(1-phenylpropyl)-2,5-imino-D-
glucitol (33)
€
€
14. Hausler, H.; Stutz, A. E. In Topics in Current Chemistry––
Glycoscience; Stu€tz, A. E., Ed.; Springer: Berlin, 2001, 215,
p 77.
Hydrogenation of compound 43 (25 mg, 0.16 mmol)
employing Pd(OH)2/C (20%) under hydrogen atmo-
15. (a) Berger, A.; de Raadt, A.; Gradnig, G.; Grasser, M.;
sphere at ambient pressure gave product 33 (29 mg,
20
D
67%): ½aꢃ +7.1 (c 0.1, MeOH). 13C NMR (CD3OD): d
€
Low, H.; Stutz, A. E. Tetrahedron Lett. 1992, 33, 7125; (b)
Legler, G.; Korth, A.; Berger, A.; Ekhart, C.; Gradnig, G.;
€
142.1, 128.3, 128.2, 125.6 (phenyl), 79.4 (C-3), 77.8 (C-
4), 70.4 (C-7), 69.5 (C-6), 66.3 (C-2), 62.1 (C-1), 60.2 (C-
5), 32.1 (C-9), 31.5 (C-8). 1H NMR: d 7.29–7.13 (m, 5H,
phenyl), 3.95 (dd, 1H, J3;4 ¼ 4:83, H-3), 3.84 (dd, 1H,
J4;5 ¼ 7:0, H-4), 3.74–3.58 (m, 3H, H-1, H-10, H-5),
3.57–3.26 (m, 4H, H-6, H-60, H-7), 2.98 (dd, 1H,
J2;3 ¼ 2:2, H-2), 2.69 (t, 2H, H-9), 1.79 (m, 2H, H-8).
€
Stutz, A. E. Carbohydr. Res. 1993, 250, 67.
16. Prade, H.; Mackenzie, L. F.; Withers, S. G. Carbohydr.
Res. 1998, 305, 371.
17. Leatherbarrow, R. J. GraFit Version 3.0, Erithacus
Software, Staines, U.K., 1992.
18. Amato, J. S.; Karady, S.; Sletzinger, M.; Weinstock, L. M.
Synthesis 1979, 12, 970.