Ethambutol-sugar hybrids as potential inhibitors of mycobacterial cell-wall biosynthesis

Article abstract of DOI:10.1016/S0008-6215(99)00069-5

Ethambutol is an established front-line agent for the treatment of tuberculosis, and is also active against Mycobacterium avium infection. However, this agent exhibits toxicity, and is considered to have low potency. The action of ethambutol on the mycoba

Full text of DOI:10.1016/S0008-6215(99)00069-5

www.elsevier.nl/locate/carres
Carbohydrate Research 317 (1999) 164–179
Ethambutol–sugar hybrids as potential inhibitors of
mycobacterial cell-wall biosynthesisꢀ
Robert C. Reynolds *, Namita Bansal, Jerry Rose, Joyce Friedrich, William J. Suling,
Joseph A. Maddry 1
Southern Research Institute, 2000 Ninth A6enue South, Birmingham, AL 35205, USA
Received 16 June 1998; revised 1 March 1999; accepted 5 March 1999
Abstract
Ethambutol is an established front-line agent for the treatment of tuberculosis, and is also active against
Mycobacterium a6ium infection. However, this agent exhibits toxicity, and is considered to have low potency. The
action of ethambutol on the mycobacterial cell wall, particularly the arabinan, and comparison of the structure of
ethambutol with several of the cell-wall saccharides, suggested that ethambutol–saccharide hybrids might lead to
agents with a more selective mechanism of action. To this end, eight ethambutol–saccharide hybrids were synthesized
and screened against M. tuberculosis and several clinical isolates of M. a6ium. © 1999 Elsevier Science Ltd. All rights
reserved.
Keywords: Glycosyltransferases; Tuberculosis; Ethambutol
1. Introduction
The emergence of multiple-drug-resistant
strains of tuberculosis [1], and the debilitating
infections caused by several mycobacteria in
immunocompromised individuals [2], have fu-
eled a resurgence of interest in developing
First reported over 30 years ago [7], exten-
novel drugs to combat these organisms [3–5].
sive structure–activity studies by Wilkinson
One of the primary agents used for the treat-
and coworkers established the parameters nec-
ment of tuberculosis, and one of the few drugs
essary for effective antimycobacterial activity
employed against Mycobacterium a6ium infec-
[8]. Among these prerequisites are two basic
nitrogen atoms optimally separated by a two-
tion, is (S,S)-(+)-2,2%-(ethylenediimino)di-1-
butanol, or ethambutol (1) [6].
carbon bridge, hydroxyl groups on each distal
nitrogen substituent, and, where substitution
of these terminal substituents introduces chi-
rality, the S stereochemistry at the carbon
center. These requirements are consistent with
a mechanism of action involving metal chela-
tion, and indeed antituberculosis activity par-
allels the stability of certain metal–
ꢀ A preliminary account of this work has appeared: J.A.
Maddry, W.J. Suling, R.C. Reynolds, Glycosyl transferases as
targets for inhibition of cell-wall synthesis in M. tuberculosis
and M. a6ium, Res. Microbiol., 147 (1996) 106–112.
* Corresponding author. Fax: +1-205-581-2726.
E-mail address: reynolds@sri.org (R.C. Reynolds)
1 Also corresponding author.
0008-6215/99/$ - see front matter © 1999 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(99)00069-5

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
165
Scheme 1.
ethambutol derivative chelates [8]. Beyond the
metal chelation hypothesis, however, the pre-
cise mechanism of action of 1 and its congeners
remains unknown, although the strict stereo-
chemical preference argues for a macromolecu-
lar site of action. Not until the pioneer-ing work
of Takayama et al. was the cell wall recognized
as the probable target of these compounds [9].
Subsequent investigations have shown that pre-
vention of mycolylation discovered by
Takayama resulted from inhibition of an earlier
stage of wall biogenesis, in particular arabinan
metabolism [10].
Interest in the design of inhibitors of cell-wall
biosynthesis continues not only because of the
unusual nature of this essential mycobacterial
structure, which affords many opportunities for
selective drug development, but also because
some wall inhibitors potentiate the activity of
otherwise inactive agents [11]. The possibility
that 1 exerts its antimycobacterial effect by
binding at the substrate site of a metal-utilizing,
sugar-processing enzyme, such as an arabinosyl
transferase, prompted us to examine several
conformationally constrained arabinofuranose
congeners that incorporate 1 as a substructure.
As to be expected for a long-chain acyclic
molecule, molecular modeling studies2 show
that ethambutol is very flexible, with many
energetically readily accessible conformations
likely to be populated. The modeling studies
demonstrated that several of these minima
superpose quite well upon the reference struc-
ture, a- -arabinofuranosyl phosphate, sup-
D
porting the plausibility of an arabinan site of
action. Based on this association, we synthe-
sized several arabinose and imino-arabinose
analogues to test this conjecture. We also
prepared several trehalose analogues bearing a
related chelating side chain because of the
reported link between 1 and glucose and myco-
late biosynthesis [10], and since the mycolyl
transferase might also contain an essential
cation in the active site.
2. Results and discussion
Chemistry
An ethambutol
D-arabinose hybrid. The 5-
substituted ethambutol–arabinose hybrid 4
was prepared as a diastereomeric mixture by
reductive amination of methyl 5-amino-5-de-
oxy-a-b- -arabinofuranoside (3) with 1-hy-
D
droxy-2-butanone in the presence of NaBH3CN
(Scheme 1). The amino saccharide was pre-
pared by the catalytic reduction of the known
5-azido derivative 2 [12] of
D
-arabinose, using
2 Complete conformational searches for both 1 and a-
D-
palladium on activated carbon.
arabinofuranosyl phosphate were performed using the tor-
sional grid algorithm and the MACROMODEL V. 4.5 software
suite (W.C. Still, et al. Columbia University, NY); for 1,
energy minimization to convergence was conducted with the
MM2 molecular mechanics force field, while the AMBER
force field provided in MACROMODEL was used for the ara-
binose derivative.
Ethambutol pyrrolidine hybrids. The N-alky-
lated pyrrolidine 5 was prepared and depro-
tected to form 6 using the method of Baxter and
Reitz (Scheme 2) [13], and was then reprotected
by a benzyloxycarbonyl (Cbz) group to yield 7.

166
R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
Subsequent treatment with acetone in the
presence of 2,2-dimethoxypropane and an acid
catalyst afforded the isopropylidene derivative
8, the common intermediate for both 15 and
22.
glucitol (22) was prepared by first protecting
the intermediate as the 4,6-di-t-
8
butyldiphenylsilyl derivative 13. The isopropy-
lidene protecting group was hydrolyzed in hot
80% acetic acid to form 16, followed by iodina-
tion at the 5-position using I2 –Ph3P–imidazole
[14] to give 17. Reaction with NaN3 in anhy-
drous DMF gave 18, followed by catalytic
reduction over Raney nickel yielding 19. Re-
ductive amination with 1-hydroxy-2-butanone
and deprotection gave the target 22 as a
diastereomeric mixture.
2,5-Anhydro-6-[[1-(hydroxymethyl)propyl]-
amino]-2,5-imino- -glucitol (15) was prepared
D
by selective tosylation of 8 to form 9, followed
by displacement of the tosyl group with azide
ion yielding 10, catalytic hydrogenation over
Raney nickel gave the amine 11, reductive
amination of 11 by the action of NaBH3CN
and 2-hydroxybutanone to prepare 12, and
formation of 14 and 15 by the removal of the
isopropylidene and Cbz protecting groups, re-
spectively. The product (15) obtained was
tested as a diastereomeric mixture.
Ethambutol–anhydro-
Anhydro- -glucitol (23) was prepared by
acid-catalyzed dehydration of -mannitol us-
D
-alditol hybrids. 2,5-
D
D
ing the method of Koerner et al. [15], but was
isolated as the crystalline 4,6-di-O-benzoyl-
1,3-O-isopropylidene derivative 24 (Scheme
3A). Mild acid treatment and monotosylation
The related derivative 2,5-anhydro-1-{[1-
(hydroxymethyl)propyl]amino}-2,5-imino- -
D
Scheme 2.

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
167
Scheme 3.
was necessary to use a different strategy to
prepare the trehalose–ethambutol hybrids.
Preparation of the 6,6%-ditrityl derivative fol-
lowed by perbenzoylation and detritylation
isolated the 6,6%-positions for reaction with
triflic anhydride by the method of Baer et al.
for preparation of the analogous hexa-O-
acetyl compound (Scheme 4A) [17]. Treatment
of the 6,6%-ditriflate 39 with either (R)(−)- or
led to 2,5-anhydro-4,6-O-dibenzoyl-1-O-p-
tolylsulfonyl- -glucitol (26). Treatment with
D
NaN3 and debenzoylation followed by reduc-
tion with Raney nickel gave the desired 1-
amino compound 29 as an oil. Treatment of
the amine with 1-hydroxy-2-butanone in the
presence of NaBH3CN–HOAc afforded the
desired ethambutol–arabinose hybrid 30. This
product appeared to be formed in a 1:1 ratio
by TLC analysis, and each diastereomer could
be isolated for screening.
(S)(+)-2-amino-1-butanol in CH2
Cl at room
2
temperature (rt), afforded the diastereomers of
44 and 45, that were deprotected with
MeOH–NaOMe to yield the target trehalose–
ethambutol hybrids 46 and 47 (Scheme 4B).
The 6,6%-ditriflate was also reacted with
potassium cyanide, and reduced to give the
homologated 6,6%-diaminomethyl trehalose
analog 42 (Scheme 4A). Reductive amination
with 1-hydroxy-2-butanone in the presence of
NaBH3CN–HOAc gave the trehalose–etham-
2,5-Anhydro-
by ring contraction of
D
-mannitol (31) was prepared
D
-glucosamine hy-
drochloride via diazotization, followed by re-
duction of the resulting aldehyde with sodium
borohydride according to the procedure of
Horton and Philips (Scheme 3B) [16]. The
crude product was isolated as the tetraacetate,
which was then deprotected and monotosy-
lated at C-1 (a significant amount of the dito-
syl compound is also formed) and treated with
azide. The tetraacetate is easily chro-
matographed or distilled via Kugelrohr. This
removes the need for large quantities of
mixed-bed resin in the reported procedure.
Reduction of the azide derivative followed by
reductive amination of the amine product us-
ing NaBH3CN–HOAc and 1-hydroxy-2-bu-
tanone led to the desired ethambutol–
arabinose hybrid 35.
butol hybrid 43,
diastereomers.
a
mixture of four
Antimycobacterial acti6ity.—Compounds 4,
15, 22, 30a and 30b, 35, 43, 46, and 47 were
evaluated for antimycobacterial activity
against M. tuberculosis strain H37Ra and a
panel of three to five clinical isolates of M.
a6ium using a colorimetric microdilution
broth assay to determine the minimum in-
hibitory concentration (MIC) [4]. All of the
compounds had MICs of \128 mg/mL while
the MICs of ethambutol for the same strains
ranged from 8 to 32 mg/mL.
Trehalose–ethambutol hybrids. As a result
of the minimal nucleophilicity of 6,6%-di-
aminotrehalose (unpublished observations), it

168
R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
Scheme 4.
Temp apparatus and are uncorrected.
Experimental
Elemental analyses were performed by the
Molecular Spectroscopy Section of the South-
ern Research Institute or by Atlantic Micro-
General methods.—Melting points were de-
termined by the capillary method on a Mel-

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
169
procedure given for compound 12, and the
crude material was chromatographed with 9:1
CHCl3–MeOH containing 1% NH4OH. A
clear, colorless oil was obtained in 41% yield.
This oil was further purified over a Bio-Bead
column, dissolved in water, frozen over a dry
ice–acetone bath, and lyophilized to obtain an
lab, Inc., Atlanta, GA. Where solvents like
EtOH or water are included in the reported
analysis, their presence was supported by the
1
1H NMR spectrum. H NMR and 13C NMR
spectra were recorded on a Nicolet NT300NB
spectrometer operating at 300.635 and 75.603
MHz, respectively. Chemical shifts are re-
ported in parts per million (ppm) downfield
from Me4Si (1H NMR) and acetone (13C
NMR), and chemical shifts for multiplets are
measured from the approximate center of the
multiplet. Coupling constants (J values) are
reported in Hz. In all cases where 13C NMR
was recorded, final target compounds ap-
peared to be formed in an equal ratio of
diastereomers within the error of the peak
height measurements. Optical rotations were
measured on a Perkin–Elmer polarimeter,
model 241 for solutions in water. Mass spectra
were recorded on a Varian/MAT 311A dou-
ble-focusing mass spectrometer in the fast
atom bombardment (FAB) mode. Addition of
a trace of LiCl to some samples gave an
[M+Li]+ cluster peak that was stronger than
the [M+H]+ peak. IR spectra were recorded
on a Nicolet FT IR spectrometer, model
10DX, using KBr discs. Thin-layer chro-
matography was performed on Analtech pre-
coated (250 mm) silica gel (GF) plates.
Column chromatography in either the flash
column or gravity mode was performed with
230–400 mesh Silica Gel 60 from E. Merck.
1
analytical sample; MS: m/z 236 [M+H]+; H
NMR (CDCl3): 0.93, 0.95 (2 t, 3 H, –
CH2CH3), 1.38–1.55 (m, 2 H, –CH2CH3),
2.58 (m, –NHCH–), 2.74, 2.82 (2 dd, 1 H,
5–CH2), 3.08, 3.14 (2 dd, 1 H, 5-CH2), 3.48
(m, 1 H, –CH2OH), 3.62–3.76 (m, 1 H, –
CH2OH), 3.86, 3.89 (2 bs, 1 H, H-3), 3.96 (s,
1 H, H-2), 4.24 (m, 1 H, H-4), 4.90, 4.91 (2 bs,
1 H, H-1). Anal. Calcd for C10H21NO5: C,
51.05; H, 9.00; N, 5.95. Found: C, 51.18; H,
9.05; N, 5.94.
2,5-Anhydro-N-benzyloxycarbonyl-2,5-im-
ino- -glucitol(7).—To6(2.3g,14.72mmol)and
D
NaHCO3 (1.6 g, 19.14 mmol) taken in 1,4-
dioxane–water (200 mL, 95:5 v/v), was added
benzyl chloroformate (2.7 mL, 19.14 mmol) in
1,4-dioxane over a period of 25 min. The
mixture was stirred for 18 h at 25 °C. The
reaction mixture was then evaporated, diluted
with water and extracted with EtOAc (4×200
mL). The combined organic layers were dried
(Na2SO4), evaporated, and dried in vacuo
over P2O5 to give 3.74 g of product 7 as a
clear oil (89% yield); MS: m/z 298 [M+1]+;
1H NMR (300 MHz, Me2SO-d6): l 3.40–3.76
(m, 5 H, 5-CH2OH, 2-CH2OH, H-5), 3.82 (m,
1 H, H-2), 4.0 (m, 2 H, H-3, H-4), 4.4, 4.93
(bs, 2 H, 2 and 5 CH2OH), 5.08 (s, 2 H,
CH2Ph), 5.13, 5.24 (d, 2 H, 3-OH, 4-OH,
JH-3,HO-3 4.4 Hz), 7.36 (m, 5 H, ArH).
Methyl
5-amino-5-deoxy-h,i- -arabino-
D
furanoside (3).—Compound 3 was prepared
according to the method for compound 11.
The crude material was chromatographed on
silica gel successively with 9:1, 4:1, and 2.3:1
CHCl3–MeOH, to afford a yellow solid in
32% yield. This material was rechro-
matographed using 3.77:1 CHCl3–MeOH
with 1.5% Et3N to obtain an analytical sam-
ple. MS: m/z 164 [M+H]+; 1H NMR
(Me2SO-d6): l 2.60 (m, 1 H, H-5), 2.76 (m, 1
H, H-5%), 3.22 (s, 3 H, OCH3), 3.62 (m, 1 H,
H-3), 3.70 (m, 1 H, H-4), 3.75 (m, 1 H, H-2),
4.60 (d, 1 H, J1,2 1.8 Hz, H-1). Anal. Calcd for
C6H13NO4: C, 44.16; H, 8.03; N, 8.58. Found:
C, 44.27; H, 8.05; N, 8.45.
2,5-Anhydro-N-benzyloxycarbonyl-2,5-im-
ino-1,3-O-isopropylidene- -glucitol (8).—To
D
200 mL of acetone (dried over MgSO4) was
added 2,2-dimethoxypropane (10.5 mL, 85.4
mmol). The solution was stirred for 2 min and
70% HClO4 (9.1 mL) was added, and the
resulting solution was stirred for 5 min. The
clear solution was then added to 7 (6.37 g,
21.4 mmol). The solution was stirred for 30
min, after which NaHCO3 was added slowly.
Stirring was continued for an additional 20
min and the mixture was evaporated to re-
move the solvent. Water was added to the
Methyl 5-deoxy-5-{[1-(hydroxymethyl)pro-
pyl]amino}-h,i-
Compound 4 was prepared according to the
D
-arabinofuranoside
(4).—

170
R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
pressure and the residue was extracted with
2:1 acetone–ether. The extract was concen-
trated and purified by silica gel column chro-
matography (99:1 CHCl3 –MeOH) to give
pure 10 (2.86 g, 76% yield); MS: m/z 363
[M+H]+.
residue, and the mixture was extracted with
CHCl3 (3×100 mL). The combined extracts
were dried (Na2SO4), evaporated, and dried to
give the product 8 as a clear oil (6.1 g). The aq
layer was extracted with EtOAc to recover
unreacted starting material. The yield was
96%, based on recovered starting material;
6-Amino-2,5-anhydro-2,5-imino-N-benzyl-
1
MS: 338 [M+H]+; H NMR (300 MHz,
oxycarbonyl-1,3-O-isopropylidene- -glucitol
D
(11).—To 10 (2.86 g, 7.89 mmol) dissolved in
EtOH (100 mL) was added Raney nickel (8.6
g, wet wt). The mixture was hydrogenated at
atmospheric pressure for 4 h at 25 °C. The
resulting mixture was filtered though Celite
and the catalyst washed with EtOH. The com-
bined filtrate and wash were evaporated to
dryness and chromatographed on silica gel
(5:1 CHCl3–CH3OH+1% NH4OH) to give
pure 11 as a white foam. MS: m/z 337 [M+
Me2SO-d6): l 1.24 (s, 3 H, Me), 1.34 (bs (s at
80 °C); 3 H, Me), 3.58 (m, 2 H, 5-CH2OH),
3.6–3.9 (m, 3 H, H-2, H-5, and 1 H of 2-
CH2O), 4.03 (bs, 2 H, 1 H of 2-CH2OH, H-4),
4.16 (bs, 1 H, H-3), 4.82 (t, 1 H, 5-CH2 OH),
5.10 (bs, 2 H, CH2Ph), 5.30 (d, 1 H, 4-OH),
7.36 (m, 5 H, ArH). Anal. Calcd for
C17H23NO6·0.05 C2H5OH: C, 60.10; H, 7.07,
N, 4.06. Found: C, 60.47; H, 6.91; N, 4.12.
2,5-Anhydro-N-benzyloxycarbonyl-2,5-im-
1
H]+; H NMR (300 MHz, Me2SO-d6): l 1.24
ino-1,3-O-isopropylidene-6-O-p-tolylsulfonyl- -
D
(s, 3 H, CH3), 1.35 (bs, 3 H, CH3), 1.56 (bs, 2
H, NH2), 2.80 (bd, 2 H, 5-CH2NH2), 3.59 (t, 1
H, H-5), 3.70–4.08 (m, 4 H, H-2, 2-CH2O,
H-4), 4.13 (bs, 1 H, H-3), 5.08 (bs, 2 H,
CH2Ph), 5.24 (bs, 1 H, 4-OH), 7.36 (m, 5 H,
ArH). Anal. Calcd for: C17H24NO5·0.3 H2O:
C, 59.49; H, 7.25; N, 7.90. Found: C, 59.74,
H, 7.25; N, 8.20.
glucitol (9).—To a cold solution (−5 °C) of 8
(6.1 g, 18.1 mmol) in dry pyridine (50 mL)
was added portionwise p-toluenesulfonyl chlo-
ride (3.5 g, 18.4 mmol). The solution was
maintained for 18 h at −5 °C, after which it
was filtered and washed with additional
pyridine. The combined filtrate and wash was
poured into an ice-cold solution of NaHCO3
(18.4 mmol, 1.5 g). The aq layer was extracted
with CHCl3 (3×150 mL). The combined
CHCl3 extracts were dried (MgSO4) and evap-
orated, and immediately chromatographed on
a silica gel column (98:2 CHCl3–MeOH) to
give 5.1 g (57% yield) of pure 9. MS: m/z 492
2,5-Anhydro-N-benzyloxycarbonyl-6-deoxy-
6 - {[1 - (hydroxymethyl)propyl]amino} - 2,5-
imino-1,3-O-isopropylidene- -glucitol (12).—
D
To a solution of 11 (500 mg, 1.49 mmol) in
dry MeOH was added 1-hydroxy-2-butanone
(0.13 mL, 1.5 mmol), followed by the addition
of HOAc (0.09 mL, 1.5 mmol) and NaCNBH3
(95 mg, 1.5 mmol). The solution was stirred
for 24 h at 25 °C. The pH of the solution was
adjusted to 2 with 2 M HCl; the resulting
solution was stirred for 5 min, and the pH was
then adjusted to ꢀ8 with M NaOH. The
solvent was evaporated, water was added, and
the aq layer was extracted with CHCl3 (5×
100 mL). The combined CHCl3 layers were
dried (MgSO4), evaporated, and dried in
vacuo to give 12 (600 mg, 98% yield); MS: m/z
409 [M+H]+.
1
[M+H]+; H NMR (300 MHz, Me2SO-d6): l
1.04 (bs, 3 H, Me), 1.35 (bs, 3 H, Me), 2.40 (s,
3 H, –SO2C6H4CH3), 3.72 (bd, 1 H, 1 H of
2-CH2O), 3.78–4.18 (m, 7 H, 1 H of 2-CH2O,
H-2, H-3, H-4, 5-CH2O H-5), 5.05 (bs, 2 H,
CH2Ph), 5.57 (d, 1 H, 4-OH), 7.34 (m, 7 H,
ArH, 2 H of SO2C6H4CH3), 7.70 (d, 2 H of
SO2C6H4CH3).
2,5-Anhydro-6-azido-N-benzyloxycarbonyl-
6-deoxy-1,3-O-isopropylidene-2,5-imino- -glu-
D
citol (10).—To 9 (5.1 g, 10.37 mmol) in dry
DMF (100 mL) was added NaN3 (2.7 g, 41.5
mmol), and the mixture was heated for 24 h at
100 °C. TLC showed the reaction was com-
plete (sprayed with NBP* to see the disap-
pearance of the starting material). The
solution was evaporated under diminished
2,5-Anhydro-N-benzyloxycarbonyl-4,6-di-O-
tert - butyldiphenylsilyl - 2,5 - imino - 1,3 - O - iso-
propylidene- -glucitol (13).—To 8 (11.5 g,
D
34.2 mmol) in dry DMF (150 mL) was added
t-butylchlorodiphenylsilane (19.6 mL, 75.2
mmol), followed by the addition of imidazole

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
171
C10H22N2O4·2HCl·0.1H2O). C, 38.87; H, 7.89;
N, 9.07. Found: C, 38.67; H, 8.21; N, 8.84.
2,5-Anhydro-N-benzyloxycarbonyl-4,6-di-O-
(10.2 g, 150.4 mmol). The solution was stirred
for 24 h at rt, after which, the solvent was
removed in vacuo at ꢀ40 °C. The residue was
partitioned between EtOAc and satd NaCl
solution. The organic layer was removed and
the aq layer was extracted (3×) with EtOAc.
The combined organic layers were dried
(MgSO4) and concentrated. The crude product
was chromatographed on a silica gel column
(95:5 cyclohexane–EtOAc) to give 13 (15.6 g,
tert-butyldiphenylsilyl-2,5-imino- -glucitol
D
(16).—Compound 13 (13 g, 16 mmol) was
heated in 1:20 MeOH–80% aq AcOH (400
mL) at 50 °C (under reflux conditions) for 24
h. The reaction mixture was processed as pre-
viously described (compound 14). Column
chromatography (3:1 cyclohexane–EtOAc) af-
forded 16 (8 g) (yield 82% based on recovered
starting material); 1H NMR (300 MHz,
1
56% yield); MS: m/z 814 [M+H]+; H NMR
t
(300 MHz, Me2SOd6): l 0.82 (s, 9 H, Bu),
t
t
Me2SO-d6): l 0.82 (s, 9 H, Bu), 1.0 (s, 9 H,
0.88 (s, 3 H, Me), 1.02 (s, 9 H, Bu), 1.20 (s, 3
tBu), 3.52 (m, 2 H, 1 H of 5-CH2O, 1 H of
2-CH2OH), 3.76 (m, 2 H, 1 H of 5-CH2O, 1 H
of 2-CH2OH), 3.96 (m, 1 H, H-2), 4.10 (m, 1
H, H-5), 4.18 (t, 1 H, H-3), 4.28 (bs, 2 H, H-4,
2-CH2OH), 4.92, 5.10 (d, 2 H –CH2Ph), 5.36
(bs, 1 H, 3-OH), 7.2–7.62 (m, 25 H, ArH).
Anal. Calcd for C40H55NO6Si·0.2H2O: C,
71.04; H, 7.18; N, 1.80. Found: C, 70.80; H,
7.37; N, 1.61.
H, Me), 3.73 (m, 2 H, 15-CH2O), 3.86 (bs, 1
H, H-2), 3.91 (bs, 1 H, 2-CH2O), 4.04 (m, 1 H,
2-CH2O), 4.16 (m, 2 H, H-3, H-5), 4.45 (bs, 1
H, H-4), 4.96–5.18 (m, 2 H, –CH2Ph), 7.22–
7.60 (m, 25 H, ArH).
2,5-Anhydro-N-benzyloxycarbonyl-6-deoxy-
6 - {[1 - (hydroxymethyl)propyl]amino} - 2,5 - im-
ino- -glucitol (14).—Compound 12 (600 mg,
D
1.47 mmol) was heated in 80% AcOH (100
mL) for 20 h at 64 °C. The solvent was evapo-
rated, toluene added, and evaporated again to
remove residual acetic acid. The residue was
chromatographed on silica gel (9:1 CHCl3–
MeOH+1% NH4OH) to give pure 14 (336
mg, 62% yield); MS: m/z 369 [M+H]+.
2,5-Anhydro-6-deoxy-6-{[1-(hydroxymethyl)-
2,5-Anhydro-N-benzyloxycarbonyl-1-deoxy-
4,6 - di - O - tert - butyldiphenylsilyl - 1 - iodo - 2,5-
imino- -glucitol (17).—To a solution of 16
D
(5.0 g, 6.5 mmol) in toluene (100 mL) were
added Ph3P (6.9 g, 26.32 mmol), imidazole
(1.8 g 26.32 mmol), and iodine (5.1 g, 19.74
mmol). The mixture was heated for 2.5 h at
108 °C. To the cooled reaction mixture was
added an equal volume of a satd NaHCO3
solution. The mixture was stirred for 5 min,
followed by the portionwise addition of
iodine. When the color of iodine persisted in
the toluene layer, the mixture was stirred for
an additional 10 min. Excess iodine was re-
moved by the addition of aq Na2S2O3. The
organic layer was diluted with toluene, ex-
tracted with water, dried (MgSO4) and con-
centrated. The residue was purified by passing
though a silica gel pad (300 g of silica gel)
with 95:5 cyclohexane–EtOAc to give pure 17
propyl]amino}-2,5-imino- -glucitol (15).—A
D
solution of 14 (0.34 g, 0.91 mmol) in 20 mL
EtOH was catalytically reduced with 5% Pd
on carbon (84 mg) at atmospheric pressure
and rt for 24 h. The mixture was filtered
though Celite and the filtrate evaporated to
dryness. The free base was converted into the
hydrochloride salt with EtOH–HCl to yield
90 mg of the product 15 (32% yield); [h]2D0+
1
8.88° (c 1); MS: m/z 235 [M+H]+; H NMR
(300 MHz, Me2SO-d6): l 0.94 (s, 3 H, Me), 1.7
(m, 24, CH2CH3), 3.1 (s, 1 H, –CH–), 3.4–
3.84 (m, 9 H, 2-CH2OH, H-2, H-5, 5-CH2NH,
CH2OH), 4.04 (s, 2 H, H-3, H-4). The OH
signals were broad between 5 and 6; 13C NMR
(300 MHz, D2O): l 78.29 (C-4), 74.12 (C-3),
64.23 (C-2), 62.37, 62.26, 62.04, 61.93 (C-5,
–NHCH(CH2OH)CH2CH3), 58.32 (–NH-
CH–(CH2OH)CH2CH3), 57.35 (C-1), 45.15
(C-6), 20.71, 20.52 (–NHCH(CH2OH)-
CH2CH3), 9.27 (CH3). Anal. Calcd for:
1
(4.1 g, 80% yield); MS: m/z 774 [M+H]+; H
NMR (300 MHz, Me2SO-d6): l 0.80 (s, 9 H,
t
tBu), 9.98 (s, 9 H, Bu), 3.08 (m, 1 H, 1 H of
2-CH2I), 3.58 (m, 2 H, 1 H of 2-CH2I, 1 H of
5-CH2O–), 3.74 (m, 1 H, 1 H of 5-CH2O),
4.18 (m, 3 H, H-2, H-3, H-5), 4.48 (s, 1 H,
H-4), 4.92, 5.12 (d, 2 H, CH2Ph), 5.83 (d, 1 H,
3-OH), 7.16–7.60 (m, 25 H, ArH). Anal.

172
R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
2,5-Anhydro-4,6-di-O-tert-butyldiphenylsilyl-
1-deoxy-1-{[1-(hydroxymethyl)propyl]amino}
Calcd for C46H54INO5Si2·0.1 H2O: C, 62.37;
H, 6.17; N, 1.58. Found: C, 61.97; H, 6.33; N,
1.51.
-2,5-imino- -glucitol (21).—To a solution of
D
20 (0.45 g, 0.54 mmol) in EtOH (20 mL) was
added 5% Pd on carbon (0.11 g). The mixture
was stirred under atmospheric hydrogen for
18 h at rt. The mixture was then filtered
though Celite and washed with more EtOH.
The combined filtrate was evaporated and
dried in vacuo over P2O5 to give 0.37 g of 21
(95% yield); MS: m/z 711 [M+H]+; 1H
NMR (300 MHz, Me2SO-d6): l 0.88 (m, 3 H,
2,5-Anhydro-1-azido-N-benzyloxycarbonyl-
1-deoxy-4,6-di-O-tert-butyl-diphenylsilyl-2,5-
imino- -glucitol (18).—To 17 (2.1 g, 2.4
D
mmol) in dry DMF (20 mL) was added NaN3
(630 mg, 9.7 mmol). The reaction mixture was
heated for 24 h at 100 °C. The reaction was
worked up as described for 10. After column
chromatography (95:5 cyclohexane–EtOAc),
1
1.5 g of pure 18 was obtained (79% yield); H
t
Me), 0.9, 1.02 (s, 18 H, Bu), 1.45 (q, 2 H,
NMR (300 MHz, Me2SO-d6): l 0.82 (s, 94,
–CH2–), 2.65 (bs, 1 H, –CH–), 2.80 and 2.96
(m, 2 H, 2-CH2), 3.02–3.42 (bm, 6 H,
CH2OH, 5-CH2O, H-5), 3.49 (m, 1 H, H-2),
3.99 (m, 2 H, H-3, H-4), 7.26–7.62 (m, 20 H,
ArH). Anal. Calcd for C42H58N2O4Si2·0.9
H2O: C, 69.36; H, 8.29; N, 3.85. Found: C,
68.99; H, 8.12; N, 3.63.
t
tBu), 1.0 (s, 9 H, Bu), 3.4–3.8 (m, 4 H,
2-CH2N3 and 5-CH2O), 4.06 (bs, 2 H, H-2,
H-5), 4.18 (bs, 1 H, H-3), 4.5 (s, 1 H, H-4),
4.92, 5.10 (d, 2 H, CH2Ph), 5.82 (bs, 1 H,
3-OH), 7.12–7.64 (m, 25 H, ArH). Anal.
Calcd for C46H54N4O5Si2: C, 69.14; H, 6.81;
N, 7.01. Found: C, 69.18; H, 7.01; N, 6.66.
1-Amino-2,5-anhydro-N-benzyloxycarbonyl-
1-deoxy-4,6-di-O-tert-butyldiphenylsilyl-2,5-
2,5-Anhydro-1-deoxy-1-{[1-(hydroxymethyl)-
propyl]amino}-2,5-imino- -glucitol (22).—To
D
a solution of 21 (0.37 g, 0.52 mmol) in ace-
tonitrile (5 mL) was added Et4NF (0.22 g, 1.5
mmol) and the solution was stirred for 24 h at
rt and then evaporated to dryness. To the
residue, THF was added and the mixture kept
in a freezer for 2 h. The THF layer was
removed and evaporated to dryness. The
residue was converted to the hydrochloride
salt with EtOH–HCl and recrystallized from
MeOH–CHCl3 to give pure 22 (33% yield);
[h]2D0+27.4° (c 0.76); MS: m/z 235 [M+H]+;
1H NMR (300 MHz, Me2SO-d6): l 0.92 (t, 3
H, Me), 1.6 (m, 2 H, –CH2CH3), 2.96 (s, 1 H
–CH–), 3.1–3.86 (m, 9 H, 2-CH2–NH, H-2,
H-5, 5-CH2OH, –CH2OH), 4.02 (s, 2 H, H-3,
H-4). The OH signals were broad between 5
and 6; 13C NMR (300 MHz, D2O): l 76.49
(C-4), 75.68 (C-3), 67.17 (C-5), 62.16 (C-2),
60.28 (C-6), 58.54, 58.46 (–NHCH(CH2-
OH)CH2CH3), 57.52, 57.48 (–NHCH(CH2-
OH)CH2CH3), 42.12, 41.95 (C-1), 20.58, 20.52
(–NHCH(CH2OH)CH2CH3), 9.29 (CH3).
Anal. Calcd for C10H22N2O4·2 HCl·0.2
C2H5OH. C, 39.48; H, 8.03; N, 8.85. Found:
C, 39.15; H, 8.31; N, 8.56.
imino- -glucitol (19).—Compound 19 was
D
made in a similar way to compound 11 except
that 2 equiv of Raney nickel (wet wt) was
used, and the reaction was stopped after 1 h
(if let run for a longer time, the Cb group
comes off). After the conventional work up
and column chromatography (9:1 cyclohex-
ane–EtOAc) 2.2 g of 19 was obtained (82%
yield based on recovered starting material); 1H
NMR (300 MHz, Me2SO-d6): l 0.82 (s, 9 H,
t
tBu), 1.0 (s, 9 H, Bu), 2.80, 2.96 (m, 2 H,
2-CH2–), 3.60, 3.74 (m, 2 H, 5-CH2O), 3.92
(m, 1 H, H-2), 4.10 (m, 1 H, H-5), 4.20 (d, 1
H, H-3), 4.46 (s, 1 H, H-4), 4.90, 5.11 (d, 2 H,
CH2Ph), 7.18–7.64 (m, 25 H, ArH). Anal.
Calcd for C46H56N2O5Si2·0.4H2O: C, 70.80; H,
7.34; N. 3.59. Found: C, 70.53; H, 7.49; N,
3.66.
2,5-Anhydro-N-benzyloxycarbonyl-4,6-di-O-
tert-butyldiphenylsilyl-1-deoxy-1-{[1-hydroxy-
methyl)propyl]amino}-2,5-imino- -glucitol
D
(20).—The procedure for making 20 was simi-
lar to that for 12 except that a 20% excess of
the reagents was added. The reaction was
monitored by MS since the starting material
and product have the same Rf value. Product
20 was obtained in 80% yield; MS: m/z 895
[M+H]+.
All of the compounds in Scheme 3 were
made from starting materials 22 and 30 by the
same standard procedures as reported in this
paper. The analytical data on the intermediate
compounds are as follows.

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
173
2,5-Anhydro-4,5-di-O-benzoyl-1,3-O-iso-
propylidene- -glucitol (24).—The 1,3-O-iso-
with 99:1 CHCl3–MeOH to afford the pure
product in 93% yield; MS: m/z 372 [M+H]+;
1H NMR (Me2SO-d6): l 3.60 (m, 1 H, 1-
CH2OH), 3.90 (m, 1 H, 1-CH2OH), 4.05 (m, 1
H, H-2), 4.28 (m, 2 H, H-3, 5-H), 4.48 (m, 2
H, 6-CH2O–), 4.68 (t, 1 H, 1-CH2OH), 5.22
(d, 1 H, H-4), 5.55 (d, 1 H, 3-OH), 7.45, 8.0
(m, 10 H, ArH).
D
propylidene derivative, prepared by a slight
modification of the known method [15], was
isolated as the crystalline 4,6-dibenzoyl com-
pound. Impure 2,5-anhydro- -glucitol [15]
D
(22.4 g, 0.14 mol) was dissolved in 500 mL dry
acetone, and dimethoxypropane (50 mL) and
conc H2SO4 (5 mL) were added with warming
to dissolve the oil. The reaction was stirred
overnight at rt, and TLC (9:1 CHCl3–MeOH)
showed complete conversion into a less polar
material. Next, the reaction was neutralized
by adding 50 mL of satd NaHCO3, and the
volatile solvents were removed by rotary evap-
oration. Solid was filtered from the remaining
solution, which was then extracted with
EtOAc (3×500 mL). Removal of the solvent
yielded 13.6 g of a colorless oil (48%) that was
carried on to the next step. The crude oil from
the previous reaction was dissolved in dry
pyridine (500 mL) and cooled on an MeOH–
ice bath to −10 °C. To this solution was
added BzCl (50 mL) dropwise with stirring.
The mixture was stirred overnight and the
reaction was stopped by pouring into ice-wa-
ter, stirring for 3 h to decompose excess BzCl,
and extracting with EtOAc (3×500 mL). The
organic layer was evaporated to remove excess
pyridine. The crude oil was dissolved in
EtOAc (500 mL), extracted with ice-cold 10%
HCl (3×100 mL) to remove excess pyridine,
followed by NaHCO3 (100 mL), and deionized
water (100 mL). The separated organic layer
was dried (Na2SO4), and evaporated to yield a
clear, colorless oil that was further purified by
column chromatography on silica gel using
CHCl3 as the elution solvent. Evaporation of
the appropriate fractions gave 25.8 g of a
clear, colorless oil (92.4%). A sample crystal-
lized readily from MeOH for spectral analysis;
2,5-Anhydro-4,5-di-O-benzoyl-1-O-p-tolyl-
sulfonyl- -glucitol (26).—Compound 26 was
D
prepared by the same procedure used for com-
pound 9. Compound 25 (0.835 g, 2.24 mmol)
was dissolved in dry pyridine (25 mL) and
cooled to −15 °C in an ice–MeOH bath.
p-Toluenesulfonylchloride (0.45 g, 1.05 equiv)
was added, and the solution was placed in a
freezer for 2 days, and then poured into ice-
water (250 mL), extracted with EtOAc (3×50
mL), 10% HCl (3×100 mL), and water (2×
100 mL) and dried (Na2SO4). After evapora-
tion, the crude material was chromatographed
on silica gel with hexanes to hexanes–EtOAc
(1:1), yielding 1.017 g (86%) as crystals from
1
MeOH; MS: m/z 527 [M+H]+; H NMR
(Me2SO-d6): l 2.38 (s, 3 H, Ar–CH3), 4.08 (m,
1 H, 1-CH2OH), 4.20 (m, 4 H, 1-CH2–, H-
2,3,5), 4.40 (m, 2 H, 6 CH2), 5.22 (m, 1 H,
H-4), 5.88 (d, 1 H, 3-OH), 7.42–8.0 (m, 34 H,
ArH).
2,5-Anhydro-1-azido-4,5-di-O-benzoyl-1-de-
oxy- -glucitol (27).—Compound 27 was pre-
D
pared by the same procedure as compound 10.
After column chromatography of the crude
material in 3:1 cyclohexane–EtOAc, 27 was
obtained in 72% yield; MS: m/z 398 [M+
1
H]+; H NMR (Me2SO-d6): l 3.52 (m, 2 H,
1-CH2OH), 4.25 (m, 1 H, H-2), 4.34 (m, 2 H,
H-3, 5-H), 4.52 (m, 2 H, 6-CH2O–), 5.24 (m,
1 H, H-4), 5.88 (d, 1 H, 3-OH), 7.2–8.02 (m,
30 H, ArH).
2,5-Anhydro-1-azido-1-deoxy- -glucitol
D
1
MS: m/z 413 [M+H]+; H NMR (Me2SO-
(28).—Compound 28 was prepared by the
same procedure used for compound 46.
Column chromatography on crude material in
9:1 CHCl2–MeOH gave 28 in 89% yield; MS
d6): l 1.30 (s, 3 H, Me), 1.44 (s, 3 H, Me), 3.90
(m, 1 H, 1-CH2OH), 4.0 (bs, 1 H, H-2), 4.1
(m, 1 H, 1-CH2OH), 4.38 (m, 1 H, 5-H), 4.55
(m, 3 H, H-3, 6-CH2O–), 5.22 (d, 1 H, H-4),
7.45–8.0 (m, 10 H, ArH).
1
m/z 190 [M+H]+; H NMR (Me2SO-d6): l
3.35 (m, 2 H, 1-CH2–), 3.45 (m, 2 H, 6-
CH2OH), 3.64 (m, 1 H, H-2), 3.82 (m, 2 H,
H-3, H-4), 3.88 (m, 1 H, 5-H), 4.90 (t, 1 H,
6-OH), 5.14 (d, 1 H, 3-OH), 5.20 (d, 1 H,
4-OH). Anal. Calcd for C6H11N3O4: C, 38.10;
2,5-Anhydro-4,5-di-O-benzoyl- -glucitol
D
(25).—Compound 25 was prepared by the
same procedure used for compound 14. Crude
material was chromatographed on silica gel

174
R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
(m, 2 H, –CH2NH), 3.34 (m, 2 H, –CH2OH),
3.48 (m, 2 H, 5-CH2OH), 3.58 (m, 1 H, H-5),
3.78 (t, 1 H, H-4), 3.84 (m, 1 H, H-3), 3.90 (m,
1 H, H-2), 5.10 (d, 1 H, 3-OH); 13C NMR
(300 MHz, D2O): l 85.00 (C-5), 79.13, 78.50,
77.91 (C-2, C-3, C-4), 62.10 (C-6, –NHCH-
(CH2OH)CH2CH3), 60.28 (–NHCH(CH2-
OH)CH2CH3), 45.40 (C-1), 22.69 (–NHCH-
(CH2OH)CH2CH3), 9.55 (CH3). Anal. Calcd
for C10H21NO5·1.4 H2O: C, 46.10; H, 9.21; N,
5.38. Found: C, 45.95; H, 9.21; N, 5.61.
H, 5.86; N, 22.21. Found: C, 38.34; H, 5.92;
N, 22.10.
1-Amino-2,5-anhydro-1-deoxy- -glucitol
D
(29).—Compound 29 was prepared by the
same procedure used for compound 11. After
evaporation of the filtrate, the residue was
triturated with ether, dissolved in water,
frozen, and lyophilized, to obtain the product
1
in 70% yield; MS: m/z 164 [M+H]+; H
NMR (Me2SO-d6): l 2.70 (dd, 1 H, 1-CH2–),
2.80 (dd, 1 H, 1-CH2 –), 3.45 (m, 2 H, 6-
CH2OH), 3.60 (m, 1 H, 5-H), 3.82 (m, 1 H,
H-4), 3.84 (m, 1 H, H-2), 3.88 (m, 1 H, H-3),
–OH and NH2 were broad. Anal. Calcd for
C6H13NO4·0.6 H2O: C, 41.42; H, 8.23; N, 8.05.
Found: C, 41.07; H, 8.39; N, 7.71.
2,5-Anhydro-1-O-p-tolyl-sulfonyl- -manni-
D
tol (32).—Compound 32 was prepared by the
same procedure used for compound 9. MS:
1
m/z 319 [M+H]+; H NMR (Me2SO-d6): l
2.22 (m, 3 H, tosyl-CH3), 3.38, 3.34 (m, 2 H,
6-CH2OH), 3.55 (m, 1 H, 5-H), 3.70 (t, 1 H,
H-3), 3.78 (m, 2 H, H-2, H-4), 4.04 (m, 2 H,
1-CH2O–), 4.74 (t, 1 H, 6-CH2OH), 5.20 (d, 1
H, 4-OH), 5.31 (d, 1 H, 3-OH), 7.50, 7.78 (d,
4 H, ArH). Anal. Calcd for C13H17O7·0.5H2O:
C, 47.84; H, 5.56. Found: C, 48.21; H, 5.94.
2,5-Anhydro-1-deoxy-1-{[1-(hydroxymethyl)-
propyl]amino}- -glucitol hydrate (30).—Com-
D
pound 30 was prepared by the same procedure
used for compound 12. Chromatography of
the
mixture
(3:1
CHCl9–MeOH+2%
NH4OH) showed two closely running prod-
ucts in ꢀ1:1 ratio, assumed to be the
diastereomers formed in the reductive amina-
tion. Column chromatography of the mixture
with the same solvent system gave two prod-
ucts 30a (35 mg) and 30b (28 mg) in overall
44% yield. The isolated ratio is an artifact of
the column because the first spot trailed into
the second making isolation of the second
material more difficult.
2,5-Anhydro-1-azido-1-deoxy- -mannitol
D
(33).—Compound 33 was prepared by the
same procedure as compound 10. MS: m/z
1
190 [M+H]+; H NMR (Me2SO-d6): l 3.32
(m, 1 H, 1-CH2–), 3.42 (m, 2 H, 1-CH2–,
6-CH2OH), 3.50 (m, 1 H, 6-CH2OH), 3.65 (m,
1 H, 5-H), 3.78 (m, 3 H, H-2, H-3, H-4), 4.72
(t, 1 H, 6-CH2OH), 5.20 (d, 1 H, 4-OH), 5.28
(d,
1
H, 3-OH). Anal. Calcd for
Compound 30a: [h]2D0 +9.11° (c 0.96); MS:
C6H11N3O4·0.1H2O: C, 37.74; H, 5.91; N,
22.00. Found: C, 37.47; H, 6.03; N, 22.17.
1
m/z 236 [M+H]+; H NMR (Me2SO-d6): l
0.82 (t, 3 H, –CH2CH3), 1.36 (m, 2 H, –
CH2CH3), 2.42 (m, 1 H, –NHCH), 2.74, 2.90
(m, 2 H, –CH2NH), 3.25 (m, 2 H, –CH2OH),
3.46 (m, 2 H, 5-CH2OH), 3.58 (m, 1 H, H-5),
3.74 (t, 1 H, H-4), 3.84 (m, 1 H, H-3), 3.90 (m,
1 H, H-2), 5.10 (d, 1 H, 3-OH); 13C NMR
(300 MHz, D2O): l 85.10 (C-5), 78.94, 78.54,
78.03 (C-2, C-3, C-4), 62.09, 62.01 (C-6, –
NHCH(CH2OH)CH2CH3), 60.18 (–NHCH-
(CH2OH)CH2CH3), 45.15 (C-1), 22.72 (–NH-
CH(CH2OH)CH2CH3), 9.63 (CH3). Anal.
Calcd for C10H21NO5·0.8 H2O: C, 48.10; H,
9.12; N, 5.61. Found: C, 47.84; H, 9.38; N,
5.65.
1-Amino-2,5-anhydro-1-deoxy- -mannitol
D
(34).—Compound 34 was prepared by the
same procedure used for compound 11; MS:
1
m/z 164 [M+H]+; H NMR (Me2SO-d6): l
2.60 (dd, 1 H, 1-CH2–), 2.70 (dd, 1 H, 1-
CH2–), 3.45 (m, 2 H, 6-CH2OH), 3.62 (m, 2
H, H-2, 5-H), 3.74 (m, 2 H, H-3, H-4); OH
and NH2 signals were broad. Anal. Calcd for
C6H13NO4·0.1H2O: C, 40.73; H, 8.35; N, 7.79.
Found: C, 41.05; H, 8.07; N, 7.53.
2,5-Anhydro-1-deoxy-1-{[1-(hydroxymethyl)-
propyl]amino}-
Compound 35 was prepared by the same pro-
cedure as compound 12. Column
D-mannitol hydrate (35).—
Compound 30b: [h]2D0 +23.6° (c 0.70); MS:
1
m/z 236 [M+H]+; H NMR (Me2SO-d6): l
chromatography using 3:1 CHCl3–MeOH +
5% NH4OH gave pure product in 50% yield;
[h]2D0+46.6° (c 1.98); MS: m/z 236 [M+H]+;
0.83 (t, 3 H, –CH2CH3), 1.36 (m, 2 H, –
CH2CH3), 2.48 (m, 1 H, –NHCH), 2.72, 2.85

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
175
[M+Li]+; 1H NMR (Me2SO-d6): l 2.58
(dd, 2 H, J5,6 2.0, J6,6 11.0 Hz, H-6), 2.86 (d,
2 H, J 11.0 Hz H-6%), 4.14 [app. d (poorly
resolved; may be dt), 2 H, J4,5 9.8 Hz, H-4],
5.68 (m, 4 H, H-2 overlapped by H-4), 5.82
(d, 2 H, J1,2 3.9 Hz, H-1), 6.07 (dd, 2 H, J2,3
10.0, J3,4 9.9 Hz, H-3), 7.16–7.84 (4 m, 60
H, Bz and trityl CH). Anal. Calcd for
C92H74O17: C, 76.12; H, 5.14. Found: C,
76.40; H, 5.16.
1H NMR (Me2SO-d6): l 0.84 (t, 3 H, –
CH2CH3), 1.40 (m, 2 H, –CH2CH3), 2.58
(m, 1 H, –NHCH–), 2.80 (m, 2 H, –
CH2NH–), 3.30 (m, 2 H, –CH2OH), 3.42
(m, 2 H, 5-CH2OH), 3.70 (m, 2 H, H-4,
H-5), 3.78 (m, 2 H, H-2, H-3); 13C NMR
(300 MHz, D2O): l 83.40, 83.36, 83.30,
81.79, 81.64 (C-2, C-3), 79.39, 77.21 (C-4,
C-5),
62.25,
62.18,
61.65
(C-6,
–NHCH(CH2OH)CH2CH3), 60.27, 60.08
(–NHCH(CH2OH)CH2CH3), 48.17, 47.97
2,3,4,2%,3%,4%-Hexa-O-benzoyl-h,h-trehalose
(38).—The mixture of 37 and the
monotrityl-hepta-O-benzoyltrehalose (93.7 g,
64.5 mmol as 37) just referred to, was dis-
solved in a mixture of MeOH (2 L) and
CH2Cl2 (1 L), and solid p-toluenesulfonic
acid was added in portions with stirring un-
til a few drops shaken with a few drops of
water gave a reading of ꢀpH 4 on Hydrion
paper. The solution was heated to reflux and
then cooled slowly and stirred for 3 days at
rt. Triethylamine (5 mL) was added; solvents
were evaporated, and a solution of the
residue in CHCl3 (500 mL) was washed by
shaking with water (500 mL) to which was
added 10 mL of 1 M HCl, satd NaHCO3
solution (500 mL), and water (500 mL). The
dried (Na2SO4) organic layer was evaporated
to give 112.6 g of a pale amber gum.
Column chromatography on silica gel with
elution by CHCl3–MeOH (99:1) gave
Ph3COCH3 (33.9 g) and Ph3COH (1.0 g).
Further elution with 98:2 gave, in order, the
hepta-O-benzoyltrehalose (11.0 g, ꢀ16%),
the desired hexa-O-benzoyltrehalose 3 (7.31
g, ꢀ12%) slightly contaminated with trail-
ings from the heptabenzoyl band, and homo-
geneous 3 (40.31 g, 64%) as a white solid.
After drying in vacuo over P2O5 at rt the
solid had mp ꢀ120 °C with prior sintering;
MS: m/z 967 [M+H]+, 845 [M−Obz]+,
475 (1/2[M−16])+, with LiCl, 973 [M+
(C-1),
22.84,
22.73
(–NHCH–
(CH2OH)CH2CH3), 9.61 (CH3). Anal. Calcd
for C10H21NO5·0.8 H2O: C, 48.10; H, 9.12;
N, 5.61. Found: C, 47.81; H, 9.02; N, 5.92.
2,3,4,2%,3%,4%-Hexa-O-benzoyl-6,6%-di-O-tri-
phenylmethyl-h,h-trehalose (37).—A solution
of a,a-trehalose dihydrate (26.95 g, 71.23
mmol) in pyridine (1500 mL) was distilled
slowly until the vapor temperature rose to
115 °C and the poorly soluble anhydrous tre-
halose began to separate. The slurry was
cooled to 10 °C, and chlorotriphenylmethane
(41.33 g, 148 mmol) was added. The stirred
mixture was allowed to warm slowly to rt.
After 4 days, the bright yellow solution was
again chilled to near 0 °C; BzCl (90.1 g, 641
mmol) was added rapidly dropwise, and the
solution was stirred for 2 days at rt and
then poured with vigorous stirring into ice-
water (8 L). A solution of the solid preci-
pitate in CH2Cl2 (1 L) was washed by shaking
with water, satd NaHCO3, and water (500
mL of each). Evaporation of the dried
(Na2SO4) organic layer gave a brittle foam
that was recrystallized from EtOH (6 L) to
give an off-white waxy solid that was dried
in vacuo over P2O5 at rt; yield 66.2 g (64%).
Concentration of the filtrate gave additional
product suitable for use as an intermediate;
yield 28.32 g (27%). Both crops contained a
mixture of 2 with a small amount of
2,3,4,6,2%,3%,4%-hepta-O-benzoyl-6-O-trityl-a,a-
trehalose (estimated B20% by TLC). To ob-
tain a sample of 2 for analysis, a small
portion of the main crop was recrystallized
twice from EtOH and dried in vacuo over
P2O5 for 4 h at 65 °C; mp 136–137 °C with
prior sintering; MS: m/z 1351 (M+H)+,
1191 [M−Ph3CO]+, 717 (1/2[M−16])+,
243 [Ph3C]+, 105 [Bz]+; with LiCl, 1457
1
Li]+; H NMR (CDCl3): l 2.45 (dd, 2 H,
6-OH), 2.92, 3.14 (2m, 4 H, H-6,6), 3.90 [d
(poorly resolved; may be dt), 2 H, J 10.0
Hz, H-5], 5.40 (dd, 2 H, J1,2 3.9, J2,3 10.1
Hz, H-2), 5.48 (dd, 2 H, J3,4 =J4,5 =9.9 Hz,
H-4), 5.71 (d, 2 H, J1,2 3.9 Hz, H-1), 6.31
(dd, 2 H, H-3), 7.32–8.05 (4m, 30, Bz).
Anal. Calcd for C54H46O17·0.5 H2O: C,
66.46; H, 4.85. Found: C, 66.55; H, 4.84.

176
R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
MS: m/z 985 [M+H]+, 863 [M−OBz]+, 484
(1/2[M−16])+, with LiCl, 991 [M+Li]+; IR:
wmax 2258 cm−1 (CN, weak), 1734 (Bz CꢀO);
1H NMR (CDCl3): l 2.00, 2.06 (dd, 2 H,
H-6), 2.09, 2.15 (dd, 2, H-6%), 4.18 (dt, 2,
J5,6=J5,6%=5.1, J4,5 10.0 Hz, H-5), 5.47 (m, 4
H, H-4 and H-2), 5.73 (d, 2 H, J1,2 3.9 Hz,
H-1), 6.24 (dd, 2 H, J2,3 =J3,4 =10.0 Hz, H-3)
7.34–8.10 (m, 30, Bz). Anal. Calcd for
C56H44N2O15: C, 68.29; H, 4.50; N, 2.84.
Found: C, 68.27; H, 4.44; N, 2.84.
2,3,4,2%,3%,4%-Hexa-O-benzoyl-6,6%-di-O-tri-
fluoromethylsulfonyl-h,h-trehalose (39).—Un-
der a nitrogen atmosphere, a solution of trifl-
uoromethanesulfonic anhydride (17.35 g,
61.48 mmol) in dry CH2Cl2 (50 mL) was
added dropwise to a solution of dry pyridine
(9.73 g, 123 mmol) in CH2Cl2 (100 mL) that
had been precooled to −10 °C. The white
slurry was stirred for 15 min at −10 °C after
addition was complete, and a solution of 38
(15.00 g, 15.37 mmol) in CH2Cl2 (100 mL) was
added rapidly dropwise, while the temperature
was kept below 0 °C. The pale yellow solution
was stirred in the cooling bath for 30 min and
then transferred to a separating funnel and
washed with ice-cold dilute HCl (250 mL, 0.5
M), cold satd NaHCO3 (2×250 mL), and
cold water (2×250 mL). The dried (Na2SO4)
organic layer was evaporated to give a porous
brittle foam that was used without further
treatment; yield 18.68 g (99%); MS (with
LiCl): m/z 1237 [M+Li]+, 1160 [M−
CHF3]+, 1109 [M−OBz]+, 607 (1/2[M−
6,6%-Dicyano-6,6%-dideoxy-h,h-trehalose
(41).—A solution of NaOCH3 in MeOH (3
mL of 1.2 N) was added to a suspension of 40
(2.58 g, 2.62 mmol) in dry MeOH (75 mL).
The mixture was stirred for 1 h at rt; then the
solution was diluted with an equal volume of
water and adjusted to pH 3.5 by portionwise
addition of Dowex 50W-X8 (H+) cation-ex-
change resin. After filtration and evaporation
of the filtrate, the residue was evaporated with
EtOH (3×) to aid removal of water. The
residue was triturated with Et2O and washed
liberally on a filter funnel with Et2O to remove
MeOBz. A solution of the solid in abs EtOH
(50 mL) was diluted slowly dropwise with
Et2O (400 mL) and the off-white solid was
collected by filtration and dried briefly in
vacuo; yield 683 mg. Evaporation of the
filtrate and reprecipitation with EtOH–Et2O
(10 and 250 mL) gave a second crop of a
white solid identical by TLC with the main
crop; yield 144 mg. The crops were combined
and redried in vacuo over P2O5 for 48 h at
82 °C; total yield 804 mg (81% as
C14H20N2O9·0.25EtOH·0.4H2O); mp ꢀ125 °C
with prior sintering. Solvation was confirmed
1
16])+; H NMR (CDCl3): l 3.76 (dd, 2 H, J5,6
2.0 Hz, H-6), 3.95 (dd, 2 H, J5,6 4.0, J6,6 11.0
Hz, H-6%), 4.25 (double m, 2 H, H-5), 5.43 (dd,
2 H, J1,2 3.9, J2,3 10.0 Hz, H-2), 5.56 (dd, 2 H,
J3,4 =J4,5=10 Hz, H-4), 5.71 (d, 2 H, J1,2 3.9
Hz, H-1), 6.26 (dd, 2 H, H-3), 7.35–8.08 (4 m,
30 H, Bz).
2,3,4,2%,3%,4%-Hexa-O-benzoyl-6,6%-dicyano-
6,6%-dideoxy-h,h-trehalose (40).—Solid KCN
(1.06 g, 16.24 mmol) was added to a solution
of the ditriflate 39 (5.00 g, 4.06 mmol) in a
mixture of MeCN (90 mL) and water (10 mL),
and the resulting yellow solution was stirred
for 24 h at rt. Solvents were evaporated, and
the residue was stirred with CH2Cl2 (200 mL).
The mixture was filtered, and the filtrate was
washed by shaking gently (to prevent emul-
sion formation) with water (100 mL). The
dried (Na2SO4) organic layer was evaporated
to give a gummy yellow residue; yield 3.50 g
(87%). Column chromatography on silica gel
with 2:1 cyclohexane–EtOAc as eluent gave
the desired product as a viscous gum. Evapo-
ration with CH2Cl2 (2×50 mL) and drying
under high vacuum gave a tractable brittle
white foam that was dried further in vacuo
over P2O5 for 20 h at 65 °C; yield 2.66 g
(66%); mp (softens gradually) ꢀ116–120 °C;
1
by H NMR; [h]2D0+140.7° (c 0.56); MS: m/z
361 [M+H]+, 343 [M−OH]+; IR: wmax 2259
1
cm−1 (CN); H NMR (Me2SO-d6): l 2.76 (m,
4 H, 6-CH2), 3.04 (m, 2 H, H-4), 3.30 (m, 2 H,
H-2), 3.53 (m, 2 H, H-3), 3.92 (dt, 2 H, H-5),
4.88 (d, 2 H, J1,2 3.9 Hz, H-1), 5.00 (m, 4 H,
OH-2 and OH-3), 5.35 (d, 2 H, J 5.2 Hz,
OH-4); 13C NMR (300 MHz, D2O): l 118.91
(2×CN), (C-2, C-3, C-4, C-2%, C-3%, C-4%),
67.94 (C-5, C-5%), 20.41 (C-6, C-6%). Anal.
Calcd for C14H20N2O9·0.25 EtOH·0.4 H2O: C,
45,95; H, 5.93; N, 7.39. Found: C, 46.01; H,
5.88; N, 7.42.

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
177
with CH2Cl2 (2×25 mL). A smaller crop (44
mg, 20%) with only a trace contaminant
(TLC) was also collected using the same
workup. The homogeneous crop was dried in
vacuo over P2O5 for 18 h at 65 °C; yield 96
mg (43%); mp softens gradually above
ꢀ80 °C; [h]2D0+116.3° (c 1.15); MS: m/z 513
[M+H]+, 248 (1/2[M−16])+, 102 [EtCH-
6,6%-Bis(aminomethyl)-6,6%-dideoxy-h,h-tre-
halose dihydrochloride (42).—A mixture of
the dinitrile 41 (735 mg, 1.94 mmol), PtO2
(200 mg), CHCl3 (2 mL), and MeOH (100
mL) was hydrogenated by shaking in a Parr
apparatus at an initial H2 pressure of 50 lb/
in2. After 20 h, the mixture was filtered
though a Celite pad under N2 pressure, and
the catalyst was washed with MeOH. The
colorless filtrate was evaporated; the residue
was dissolved in dry MeOH (20 mL), and the
solution was diluted dropwise with Et2O (350
mL). The dense solid deposit was filtered un-
der N2 pressure, washed with Et2O, and dried
in vacuo over P2O5 at rt, and then for 20 h
at 82 °C; yield 797 mg (89%); mp dec above
160 °C with prior charring; MS: m/z 369
[M+H]+, 176 (1/2[M−16])+; 1H NMR-
(Me2SO-d6): l 1.59, 2.05 (complex m, 4
H, 6-CH2), 2.82 (complex m, 4 H, H2NCH2–),
2.92 (dd, 2 H, J 9.5 Hz, H-4), 3.30 (dd, 2
H, J 3.9 Hz, H-2), 3.48 (m, 2 H, H-3), 3.84
(dd, 2 H, J 3.2 Hz; H-5), 4.81 (d, 2 H, J1,2
3.9 Hz, H-1), 5.01 (d, 2 H, J 4.8 Hz, 3-OH),
5.18 (br, 4 H, 2-OH and 4-OH), 7.87 (br, 6
H, –NH3+). Anal. Calcd for C14H28N2-
O9·2HCl·H2O: C, 36.61; H, 7.02; N, 6.10; Cl,
15.44. Found: C, 36.64; H, 7.00; N, 6.02; Cl,
15.30.
1
(CH2OH)NHCH2]+; H NMR (Me2SO-d6):
l 0.84 (t, 6 H, CH3CH2), 1.39 (m, 4 H,
CH3CH2), 1.44 (m, 2 H, H-6), 1.88 (m, 2
H, H-6%), 2.45 (m, 2 H, EtCHNH–), 2.65 (m,
4 H, –NHCH2CH2), 2.90 (dd, 2 H, J3,4 =
J4,5=9.5 Hz, H-4), 3.29 and 3.39 (2m, H2O
superimposed on H-2 and –CH2OH), 3.47
(m, 2 H, H-3), 3.78 (m, 2 H, H-5), 4.79 (m, 2
H, H-1), 4.2–5.6 (very broad, best seen in
integral, NH, OH); 13C NMR (300 MHz,
D2O): l 93.98 (C-1, C-1%), 73.79, 72.89, 71.45
(C-2, C-3, C-5, C-2%, C-3%, C-5%), 71.14, 70.89
(C-4,
(CH2OH)CH2CH3),
C-4%),
61.96,
61.85
60.12
(–NHCH-
(–NHCH–
(CH2OH)CH2CH3), 42.93, 42.83 (–CH2
NHCH(CH2OH)CH2CH3), 30.46 (C-6, C-6%),
22.73,
22.51
(–NHCH(CH2OH)CH2-
CH3), 9.63 (CH3). Anal. Calcd for
C22H44N2O11·0.1 EtOH·H2O: C, 49.82; H,
8.78; N, 5.23. Found: C, 49.79; H, 8.82; N,
5.21.
(R,S) - 6,6% - Dideoxy - 6,6% - bis{[[1 - (hydro-
xymethyl)propyl]amino]methyl}-h,h-trehalose
(43).—Solid anhydrous NaHCO3 (71 mg,
0.84 mmol) was added to a stirred solution
of the diamine dihydrochloride 42 (203 mg,
0.413 mmol) in MeOH (12 mL), followed
after 10 min by 1-hydroxy-2-butanone (87
mg, 0.99 mmol, 20% excess) and glacial
HOAc (60 mg, 0.99 mmol). After 30 min,
solid NaBH3CN (62 mg, 0.99 mmol) was
added, and the resulting colorless mixture
was stirred for 4 days at rt. Volatiles were
evaporated; a solution of the residue in water
(10 mL) was allowed to stand for 30 min,
evaporated and reevaporated with MeOH
(2×20 mL). A solution of the residue in 1:1
CHCl3–MeOH was filtered and applied to a
silica gel flash column. Elution with CHCl3–
MeOH–conc NH4OH (10:10:1) gave homo-
geneous fractions (TLC) of the desired
product that were pooled, evaporated, and
reevaporated with EtOH (2×25 mL) and
(S)(+) - 2,3,4,2%,3%,4% - Hexa - O - benzoyl-
6,6% - bis[1 - (hydroxymethyl)propylamino] - h,h-
trehalose (44).—To a solution of 39 (1.14
mmol, 1.4 g) in 5 mL of dry CH2Cl2 was
added (S)(+)-2-amino-1-butanol (5.7 mmol,
0.54 mL). The resulting solution was stirred
for 18 h at rt. The reaction mixture was
directly separated on a silica gel column with
99:1 CHCl3–MeOH as eluent to obtain 867
mg of product (68% yield); mp 93–95 °C; MS:
1
m/z 1109 [M+H]+; H NMR (CDCl3): l
0.80 (t, 6 H, CH3CH2–), 1.35 (m, 4 H,
CH3CH2), 2.25 (dd, 2 H, H-6, H-6%), 2.35 (m,
2 H, EtCHNH–), 2.62 (m, 2 H, H-6, H-6%),
3.18 (m, 2 H, CH2OH), 3.48 (m, 2 H,
CH2OH), 3.90 (m, 2 H, H-5), 5.38 (m, 2 H,
H-2), 5.45 (m, 2 H, H-4), 5.98 (d, 2 H, H-1),
6.20 (t, 2 H, H-3), 7.28–8.2 (m, 30 H, Bz).
Anal. Calcd For C62H62N2O17·0.2 H2O: C,
66.92; H, 5.83; N, 2.52. Found: C, 66.52; H,
5.91; N, 2.69.

178
R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
(m, 4 H, CH3CH2 –), 2.36 (m, 2 H,
(R)(−)-2,3,4,2%,3%,4%-Hexa-O-benzoyl-6,6%-
EtCHNH–), 2.52 (dd, 2 H, H-6, H-6%), 2.63
(dd, 2 H, H-6, H-6%), 3.04 (t, 2 H, H-4), 3.20
(m, 4 H, H-2, –CH2OH), 3.36 (m, 2 H, –
CH2OH), 3.54 (m, 2 H, H-3), 3.70 (m, 2 H,
H-5), 4.42 (bs, 2 H, –CH2OH), 4.60 (bs, 2
H, OH-2), 4.78 (d, 2 H, OH-3), 4.86 (d, 2
H, H-1), 5.0 (bs, 2 H, OH-4); 13C NMR
(300 MHz, D2O): l 93.38 (C-1, C-1%), 72.63,
72.51, 71.31, 70.40 (C-2, C-3, C-4, C-5, C-2%,
C-3%, C-4%, C-5%), 62.79 (–NHCH(CH2OH)-
CH2CH3), 59.31 (–NHCH(CH2OH)CH2CH3),
47.04 (C-6, C-6%), 22.66 (–NHCH(CH2OH)-
CH2CH3), 9.63 (CH3). Anal. Calcd for
C20H40N2O11·1.5H2O: C, 46.96; H, 8.47; N,
5.48. Found: C, 46.58; H, 8.51; N, 5.41.
dideoxy - 6,6% - bis[1 - (hydroxymethyl)propyl-
amino]-h,h-trehalose (45).—Compound 45 was
prepared by the same procedure as 44 with
(R)(−)-2-amino-1-butanol. Pure product
was obtained after silica gel chromatography
in 65% yield; mp 146–148 °C; MS: m/z 1109
1
[M+H]+; H NMR (CDCl3): l 0.82 (t, 6
H, CH3CH2), 1.30 (m, 4 H, CH3CH2), 2.0
(dd, 2 H, H-6, H-6%), 2.18 (m, 2 H,
EtCHNH–), 2.35 (dd, 2 H, H-6, H-6), 3.14
(m, 1 H, CH2OH), 3.36 (m, 1 H, CH2OH),
4.0 (m, 2 H, H-5), 5.34 (dd, 2 H, H-2), 5.68
(d, 2 H, H-1), 5.85 (t, 2 H, H-4), 6.22 (t, 2
H, H-3), 7.26–8.12 (m, 30 H, Bz). Anal.
Calcd for C62H64N2O17·0.2H2O: C, 66.92; H,
5.83; N, 2.52. Found: C, 66.51; H, 5.85; N,
2.52.
Acknowledgements
(S)(+)6,6% - Dideoxy - 6,6% - bis[1 - (hydroxy-
methyl)propylamino]-h,h-trehalose
(46).—A
This research was supported by funds from
the US National Institutes of Health (NIAID
Grants U19-AI40972 and R01-AI38667).
solution of sodium metal (20 mg) in dry MeOH
(8 mL) was added to 44 (832 mg, 0.75 mmol).
After stirring the resulting solution for 1 h,
more MeOH (10 mL) was added and the pH
was adjusted to 8 with Dowex 50W-X8 (H+)
cation-exchange resin. After filtration and
evaporation of the filtrate, the residue was
separated by column chromatography (3:1
CHCl3–MeOH+5% NH4OH) to give pure 296
mg of product (82% yield); mp 104–107 °C.
[h]2D0+138.3° (c 0.5); MS: m/z 485 [M+H]+;
1H NMR (Me2SO-d6,): l 0.80 (t, 6 H,
CH3CH2–), 1.30 (m, 4 H, CH3CH2–), 2.42 (m,
2 H, EtCHNH–), 2.62 (dd, 2 H, H-6, H-6%),
2.76 (dd, 2 H, H-6, H-6%), 3.12 (t, 2 H, H-4), 3.24
(m, 4 H, H-2, –CH2OH), 3.36 (m, 2 H, –
CH2OH), 3.54 (m, 2 H, H-3), 3.78 (m, 2 H,
H-5), 4.52 (bs, 2 H, –CH2OH), 4.68 (d, 2 H,
OH-2), 4.76 (d, 2 H, OH-3), 4.88 (d, 2 H, H-1),
4.94 (bs, 2 H, OH-4). Anal. Calcd for
C20H40N2O11·H2O: C, 49.03; H, 8.35; N, 5.72.
Found: C, 48.82; H, 8.74; N, 5.55.
References
[1] World Health Organization, in Anti-tuberculosis Drug
Resistance in the World, The WHO/IUATLD global
project on anti-tuberculosis drug resistance surveillance,
1997.
[2] H. Masur, J. Infect. Dis., 161 (1990) 858–864.
[3] D.B. Young, in B.R. Bloom (Ed.), Tuberculosis, Patho-
gens, Protection, and Control, ASM Press, Washington,
DC, 1994, pp. 559–567.
[4] J.A. Maddry, N. Bansal, L.E. Bermudez, R.N. Comber,
I.M. Orme, W.J. Suling, L.N. Wilson, R.C. Reynolds,
Bioorg. Med. Chem. Lett., 8 (1998) 237–242.
[5] A.K. Pathak, Y.A. El-Kattan, N. Bansal, J.A. Maddry,
R.C. Reynolds, Tetrahedron Lett., 39 (1998) 1497–1500.
[6] L.B. Heifets, M.D. Iseman, P.J. Lindholm-Levy, Antimi-
crob. Agents Chemother., 30 (1986) 927–932.
[7] R.G. Wilkinson, R.G. Shepherd, J.P. Thomas, C.
Baughn, J. Am. Chem. Soc., 83 (1961) 2212–2213.
[8] R.G. Shepherd, C. Baughn, M.L. Cantrall, B. Goodstein,
J.P. Thomas, R.G. Wilkinson, Ann. N.Y. Acad. Sci., 135
(1966) 686–710.
[9] K. Takayama, E.L. Armstrong, K.A. Kunugi, J.O. Kil-
burn, Antimicrob. Agents Chemother., 16 (1979) 240–242.
[10] (a) K. Takayama, J.O. Kilburn, Antimicrob. Agents
Chemother., 33 (1989) 1493–1499. (b) L. Deng, K.
Mikusova, K.G. Robuck, M. Scherman, P.J. Brennan, M.
McNeil, Antimicrob. Agents Chemother., 39 (1995) 694–
701. (c) G. Silve, P. Valero-Guillen, A. Quemard, M.-A.
Dupont, M. Daffe, G. Laneelle, Antimicrob. Agents
Chemother., 37 (1993) 1536–1538.
(R)(−)-6,6%-Dideoxy-6,6%-bis[1-(hydroxy-
methyl)propylamino]-h,h-trehalose (47).—This
compound was prepared by the same proce-
dure as 46. After column chromatography,
.
pure product was obtained in 78% yield; mp
96–98 °C with glass formation; [h]2D0+115.7°
[11] (a) N. Rastogi, V. Labrousse, J.P. Carvalho de Sousa,
Curr. Microbiol., 26 (1993) 191–196. (b) N. Rastogi, K.S.
Goh, H.L. David. Antimicrob. Agents Chemother.,
(c 0.5); MS: m/z 485 [M+H]+; H NMR
1
(Me2SO-d6): l 0.81 (t, 6 H, CH3CH2–), 1.31

R.C. Reynolds et al. / Carbohydrate Research 317 (1999) 164–179
179
34 (1990) 759–764. (c) S.E. Hoffner, M. Kratz, B.
Olsson-Liljequist, S.B. Svenson, G. Kallenius, J. Antimi-
crob. Chemother., 24 (1989) 317–324. (d) S.E. Hoffner,
G. Kallenius, A.E. Beezer, S.B. Svenson, Acta Leprolog-
ica, 7 (1989) S195–S199.
[14] P.J. Garegg, B. Samuelsson, J. Chem. Soc. Perkin 1,
(1980) 2866–2869.
[15] T.A. Koerner, R.J. Voll, E.S. Younathan, Carbohydr.
Res., 59 (1973) 403–416.
[16] D. Horton, K.D. Philips, Carbohydr. Res., 30 (1973)
367–374.
[17] H.H. Baer, R.L. Breton, Y. Shen, Carbohydr. Res., 200
(1990) 377–389.
[12] L. Schmidt, E.B. Pedersen, C. Nielsen, Acta Chem.
Scand., 48 (1994) 215–221.
[13] E.W. Baxter, A.B. Reitz, J. Org. Chem., 59 (1994) 3175–
3185.
.