Synthesis of kanamycin A analogs containing a 6-amino-6-deoxyglycofuranose moiety

Article abstract of DOI:10.1016/S0008-6215(00)00184-1

Three kanamycin A analogs containing 6-amino-6-deoxyglycofuranoses have been prepared as candidates for potential activity against resistant bacteria producing 6'-N-acetyltransferase. They are 4-O-(6-amino-3,5,6-trideoxy-α-D-, -β-D-, and -β-L-erythro-hexofuranosyl)-6-O-(3-amino-3-deoxy-α-D-glucopyran osyl)-2,5-dideoxy-5-epi-5-fluorostreptamine. Structure-activity relationships of these compounds are discussed. (C) 2000 Published by Elsevier Science Ltd.

Full text of DOI:10.1016/S0008-6215(00)00184-1

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Carbohydrate Research 329 (2000) 325–340
Synthesis of kanamycin A analogs containing a
6-amino-6-deoxyglycofuranose moiety
Yoshihiko Kobayashi a, Tetsuro Ohgami b, Katsura Ohtsuki b, Tsutomu Tsuchiya a,*
a Institute of Bioorganic Chemistry, 3-34-17 Ida, Nakahara-ku, Kawasaki 211-0035, Japan
b Department of Applied Chemistry, Faculty of Science and Technology, Keio Uni6ersity, Hiyoshi, Kohoku-ku,
Yokohama 223-8522, Japan
Received 14 February 2000; accepted 28 May 2000
Abstract
Three kanamycin A analogs containing 6-amino-6-deoxyglycofuranoses have been prepared as candidates for
potential activity against resistant bacteria producing 6%-N-acetyltransferase. They are 4-O-(6-amino-3,5,6-trideoxy-a-
D
-,
-b-
D
-,
and
-b-
L
-erythro-hexofuranosyl)-6-O-(3-amino-3-deoxy-a-D-glucopyranosyl)-2,5-dideoxy-5-epi-5-
fluorostreptamine. Structure–activity relationships of these compounds are discussed. © 2000 Published by Elsevier
Science Ltd. All rights reserved.
Keywords: Kanamycin analogs; 6-Amino-3,5,6-trideoxy-
D
- or -
L
-erythro hexofuranose; 6%-Aminoacetyltransferase
alkylation or 6%-C-alkylation of kanamycin
and its analogs [5] succeeded in improving this
somewhat, but led to lowering of the intrinsic
activity. Against this background, we intended
to prepare new kanamycin analogs potentially
active against the resistant strains by replacing
1. Introduction
Kanamycins1 are aminoglycoside antibiotics
having a broad spectrum of antibacterial ac-
tivities, but they are inactivated by a variety of
resistant bacteria that produce phosphoryl-,
nucleotidyl-, and acetyl-transferases causing
modification of several key functional groups
[1]. Such chemically modified kanamycin
the 6-amino-6-deoxy- -glucopyranose residue
D
(Glc6N) of kanamycin A with a terminal-
aminated sugar designed to avoid the recogni-
tion by the 6%-N-acetyltransferases of resistant
bacteria through changing the conformation
adjacent to the 6%-amino group of Glc6N.
This study is an attempt along the foregoing
lines, and describes the synthesis of
kanamycin A analogs (29a,b) having a 6-
derivatives as 3%,4%-dideoxykanamycin
B
(dibekacin) [2], amikacin [3], and arbekacin [4]
restore the activity of the parent compounds
against most of the resistant bacteria, but for
the strains producing 6%-N-acetyltransferases,
no effective tool to avoid the acetylation has
yet been discovered. Past attempts at 6%-N-
amino-3,5,6-trideoxy-a- or -b- -erythro-hexo-
D
furanose group in place of Glc6N, expecting
that the pyranose“furanose change might
produce some biologically useful change. The
reason for the choice of a furanose moiety
lacking 3- and 5-hydroxyl groups is based on
the expectation that the absence of a 3%-OH
* Corresponding author. Fax: +81-44-7553099.
E-mail
address:
seiyuken@mx1.alpha-web.ne.jp
(T.
Tsuchiya).
1 See structures 20–21 as representative kanamycin A
derivatives used in this investigation.
0008-6215/00/$ - see front matter © 2000 Published by Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(00)00184-1

326
Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
group in the final products 29 may prevent
possible 3%-phosphorylation by the resistant
bacteria producing 3%-phosphotransferases,
and the absence of a 5%-OH group may facili-
tate the rotation around the C-5%–C-6% axis,
making the 6%-NH2 group of the new com-
pound approach a suitable position to fit the
ribosomal RNA of bacteria [6]. In natural
(9 ), acid-catalyzed methanolysis gave an
D
anomeric mixture of methyl furanosides (10).
Benzylation (to give 11), followed by hydroly-
sis with aq hydrochloric acid–acetic acid (the
use of aq hydrochloric acid–THF attempted
first gave an undesirable 4-chlorobutyl glyco-
side) gave a free sugar 12, which led to the
glycosyl chloride 13 by treatment with SOCl2.
As 13 was unstable, the stable glycosyl
fluoride analog 14 was also prepared from 12
by treatment with Et2NSF3 (DAST) [13].
kanamycins, the Glc6N group has the a-
configuration, but in our synthesis, we pre-
pared both of the a- and b anomeric - and
-hexofuranoses to examine the activity–
structure relationships in more detail.
Forthe6-O-(3-amino-3-deoxy- -glucopyran-
D
-
D
L
D
osyl)-2-deoxystreptamine moiety (3AD), to
which the foregoing furanoses were to be cou-
pled, a fluorine atom was introduced with
inversion at C-5. One reason for this modifica-
tion is because the chemical glycosylation of a
protected 3AD derivative with a glycosyl
halide usually produces undesirable 5-O-gly-
cosyl position isomers as by-products [7],
making purification of the desired 4-O-glyco-
syl products difficult. Another reason is that
the 5-epifluorination sometimes enhances the
antibacterial activity, as observed in 5-deoxy-
5-epifluoro-amikacin and -arbekacin [8].
2. Results and discussion
Synthesis of glycosyl donors.—We at-
tempted first a one-step 3,5-dideoxygenation
of 1,2-O-isopropylidene-3,5-di-O-[(methyl-
thio)thiocarbonyl] - 6 - O - trityl - a -
D
- glucofu-
ranose according to Barton et al. [9], but a
complex mixture was obtained; we thus took a
stepwise deoxygenation route. We initially at-
tempted to convert a 3-deoxy-6-O-tosyl-
ribo-hexofuranose derivative (6 ) prepared
according to Just and Lethe [10,11] by way of
–5 into its 5-xanthate by the usual proce-
D-
D
1D
D
dure (NaH, CS2, and MeI), but the undesired
5,6-epoxide was produced by removal of the
6-tosyloxy group. Imidazolylthiocarbonyla-
tion under milder basic conditions, however,
successfully gave the 5-O-thiocarbonyl deriva-
A similar synthesis was applied to the
series. 1,2;5,6-Di-O-isopropylidene-a- -gluco-
furanose [14] was converted into 3-deoxy-6-O-
tosyl- -ribo-hexofuranose (6 ) via six steps
(1 to 5 ). 5-Thiocarbonylimidazolylation of
, followed by deoxygenation (Bu3SnH–
AIBN) in toluene in one pot gave the dideoxy
derivative (8 ), which led to the 6-azido
L
L
L
L
L
L
tive 7
the presence of AIBN [9] gave the 3,5-
dideoxy-6-O-tosyl derivative 8 [12]. After
conversion of 8 into the 6-azido derivative
D
. Deoxygenation of 7
D
with Bu3SnH in
6
L
D
D
L

Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
327
tive 21, with a free HO-5 group (this position
resists acetylation or benzoylation [8,16–18]),
which was fluorinated with DAST to give the
5-deoxy-5-epifluoro derivative 22 in almost
quantitative yield. The axial deposition of
fluorine was proven from the large coupling
constants [8] (J5,F 50.5 and J4,F=J6,F 26 Hz)
derivative (9
L
). As in the
D
series, the free
sugar (12) with a 6-azido group was unstable
on storage, we attempted conversion of the
6-azido group of 9 into a tosylamido group
L
by reduction (Raney Ni) and tosylation. The
resulting N-tosyl derivative (15) was succes-
sively methanolyzed (to give 16), benzylated
(to give the 2-O,6-N-dibenzyl derivative 17),
and hydrolyzed to give the free sugar 18,
which, however, was unstable on storage, as
19
in its F NMR spectrum. After deacetylation
(to give 23), Smith degradation [19] was per-
formed giving a dialdehyde that was reduced
(sodium
borohydride)
and
hydrolyzed
with the
with SOCl2 gave the
D
analog (12). Chlorination of 18
(methanolic hydrochloric acid) to give the tri-
N-tosyl disaccharide 24. The structure was
confirmed by the absence of the 6-tosylami-
dopyranose moiety. Subsequent benzylidena-
tion of 24 (to give 25), followed by acetylation
(N-acetylimidazole in 1:9 pyridine–dimethyl-
sulfoxide [20]) gave the 2%-O-acetyl-4-hydroxy
derivative 26 (47%) together with the 4,2%-di-
O-acetyl derivative 27 (50%). Compound 27
could be recycled to 25 after deacetylation.
Glycosylation.—As preliminary couplings
of 26 and 13 under some standard conditions
gave rise to diverse b:a ratios, conditions for
optimization were first examined (Scheme 1).
The results (Table 1) showed that the ratio
ranged from 1.0 to 7.6, and no clear-cut con-
ditions to produce the a or b anomer selec-
tively were found. As we considered that both
the a and b anomers were necessary to exam-
ine the antibacterial activities of the final
products, the coupling was carried out under
the conditions of entry 7 (i:h=1), which
showed a relatively high yield, and the
anomers produced were separated by repeated
chromatography.
L
-glycosyl donor 19,
which was unstable and was used
immediately.
After Zemple´n deacetylation of 28 in 9:1
pyridine–methanol (for 28a) or 1:1 chloro-
form–methanol (for 28b) (both 28a and 28b
are scarcely soluble even in methanol), the
products were deprotected with Na in liquid
NH3–THF (Birch reduction), whereupon the
azido (to give an amino group), N-tosyl, O-
benzyl, and O-benzylidene groups were re-
moved simultaneously to give the final
products 29; in the case of 28a, however,
reduction of the 6%-azido group (partial deben-
zylation and debenzylidenation also occurred)
was initially carried out prior to the Birch
reduction (detosylation) to increase the solu-
bility of the starting material in NH3.
Synthesis of the 3AD acceptor.—Acetyla-
tion of tetra-N-tosylkanamycin A (20) [15]
gave the 2%,3%,4%,2¦,4¦,6¦-hexa-O-acetyl deriva-

328
Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
In the coupling of 26 and 19, the best
ments of these synthetic products were per-
formed by NMR spectroscopy (Tables 2 and
3).
conditions among those tested were
Hg(CN)2–s-collidine in dichloromethane, and
30 was obtained in low yield as an anomeric
mixture (b:aꢀ3), from which the b anomer
(30b) could be separated by chromatography
(Scheme 2). Deprotection as described for 28b
gave the final product 32b. Structural assign-
Interestingly, in the 13C spectra, compound
32b with a b-
bles in its chemical shifts, 29a (a-
in the 2-deoxystreptamine (DST) moiety; on
the other hand, 32b resembles 29b (b- struc-
L
-glycofuranose moiety, resem-
D
structure)
D
Scheme 1.
Table 1
Glycosylation of 26 with 13 (or 14 a) to give 28
Entry Reagent (mol equiv b)
Solvent (v/w b)
Temperature
Time (h)
Yield (%)
b:a ratio c
1
2
3
4
5
6
7
8
9
10
11
12
13
Hg(CN)2 (2.0)
Hg(CN)2 (2.0)
Hg(CN)2 (2.0)
Hg(CN)2 (2.0)
Hg(CN)2 (2.0)
Hg(CN)2 (2.0)
Hg(CN)2 (2.0)
CH2Cl2 (5)
rt
rt
1
1
5
1
1
1
1
7
1
1
5
20
5
35
51
6.0
2.7
CH2Cl2 (10)
CH2Cl2 (10)
CH2Cl2 (50)
CH3CN(10)
CH3NO2 (10)
THF (10)
CH2Cl2 (10)
CH2Cl2 (10)
CH2Cl2 (10)
CH2Cl2 (10)
1:1 CH2Cl2–Et2O (10)
CH2Cl2 (10)
−50“0 °C
rt
rt
rt
rt
rt
rt
0 °C
no reaction
45
44
44
58
53
55
41
47
41
51
2.8
2.4
2.0
1.0
1.5
1.4
2.1
7.6
2.0
6.1
Hg(CN)2 (2.0)–s-collidine d (4.0)
HgBr2 (2.0)–HgO (4.0)
AgOTf (1.2)–TTBP e (1.2)
Ag2CO3 (2.0)
AgClO4 (3.0)–SnCl2 (3.0)
CdCO3 (2.0)
rt
−50 °C
rt
a Entry 12.
b Based on 13 (or 14).
1
c The anomeric ratios were determined based on the H NMR spectra of 28 (a anomer: H-1% l 5.45, J1%,2% 4 Hz; b anomer: H-1%
l 5.63, singlet).
d 2,4,6-Trimethylpyridine.
e 2,4,6-Tri-t-butylpyridine.

Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
329
Scheme 2.
ture) in the furanose moiety. This suggests
that the electronic states for carbons of the
DST portions of 32b and 29a resemble each
other, but differ in the furanose portions, and
for the furanose portion, the states of 32b and
29b resemble each other.
Antibacterial acti6ity.—The three com-
pounds (29a,b, and 32b) synthesized showed
no antibacterial activity (MIC\100 mcg/mL)
against normal and resistant strains tested
(32a cannot be measured due to the small
amount obtained).
trudes from the location of the 6%-amino group
of kanamycin A by more than the diameter of
one carbon; the b-
L
isomer 32b has a short
,
6%-N(32b)–6%-N(KMA) distance (1.02 A) (see
Fig. 1, stereoview). It has been suggested that
the position of the 6%-NH2 group of the
kanamycin A molecule is most important for
manifesting antibacterial activity [6].
neamine analog having a 7-amino-7-deoxy
sugar moiety (with an a- -glycopyranose
structure) instead of the 6-amino-6-deoxy-
A
D
D-
glucose moiety of kanamycin A, however, re-
tains its antibacterial activity [21], although
only weakly. Therefore, the lack of activity of
our synthetic products must be ascribed to the
steric difference between the pyranose ring (in
kanamycin A) and the furanose, suggesting
that the latter structure hinders binding of the
synthetic products to the ribosomal RNA of
bacteria [6].
Discussion.—In order to search for the un-
derlying reason for the inactivity of the prod-
ucts prepared, their energy-minimal confor-
mations (MOPAC with mimic water effect)
were calculated and compared with that of
kanamycin A. As shown in Fig. 1, where the 6-
O-(3-amino-3-deoxy-a- -glucopyranosyl)-
D
2-deoxystreptamine portions of kanamycin A
and 29a (29b, 32a, or 32b) are brought to-
gether to be almost superimposed, the spatial
dispositions of the 6-amino-3,5,6-trideoxyfura-
3. Experimental
nose and 6-amino-6-deoxy- -glucose moieties
D
are considerably deviant. A characteristic fea-
ture is the fact that the 6-amino group in each
furanose moiety, except for that of 32b, pro-
General methods.—Melting points were de-
termined on a Kofler block and are uncor-
rected. Optical rotations were determined with

/
329
(
325
340
Fig. 1. The energy-minimal conformations of 29a, 29b, 32a, and 32b computer-calculated (for details, see Section 3) for their glycofuranosyl-DST portions (dotted line),
drawn with the 4-O-(6-amino-6-deoxy-a- -glucopyranosyl)-2-deoxystreptamine moiety of kanamycin A (solid line), together with the stereoviews of 32b and kanamycin A
(both solid line) for the corresponding portions.
D

Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
331
a Perkin–Elmer 241 polarimeter. IR spectra
were recorded with a Jasco A-202 grating
spectrophotometer. NMR spectra (1H at 250
and 500 MHz, 13C at 125.8 MHz, and 19F at
235.4 MHz) were recorded with Bruker AC-
250P and AMX-500 spectrometers, using
Me4Si and CFCl3 (for 19F) as the internal
references, respectively. If necessary, signal as-
signments were performed by aid of shift-cor-
related 2D spectra. Asterisks attached after
the NMR acronym indicate that only key
signals are shown. Thin-layer chromatography
(TLC) was performed on Kieselgel 60 F254 (E.
Merck), high-performance thin layer chro-
matography (HPTLC) on Kieselgel 60 F254 for
nano-TLC (E. Merck), and column chro-
Table 2
1H NMR data for compounds 28a, 28b (pyridine-d5) and 29a, 29b and 32b (26% ND3 in D2O)
28a
28b
29a
29b
32b
H-1
3.99–4.06m
3.93–4.02m
1.96q
3.10dddd
J1,2ax 12.5, J1,6 10.5, J1,2eq 4.5, J1,F
1.10q
3.09dddd
3.13dddd
1.16q
2
H-2ax 1.93q
1.13q
2ax,2eq=J2ax,3
J
1,2ax=J2ax,2eq=J2ax,3 13
J
12.5
H-2eq 2.82dt
2.81dt
2.00dt
2.02dt
2.05dt
J
1,2eq=J2eq,3
4
J2eq,3 4.5
2.98dddd
3.44ddd
H-3
H-4
3.92–3.99m
4.24ddd
J3,4 10.5, J4,5 2, J4,F 28
5.69dt
J5,6 2, J5,F 51.5
4.25ddd
J1,6 10.5, J6,F 28
5.45d
4.02–4.12m
4.13br dd
J3,4 10.5, J4,F 27
5.67br d
3.03dddd
3.47ddd
3.01dddd
3.53ddd
J3,4 10.5, J4,5 2, J4,F 29
H-5
H-6
H-1%
H-2%
5.22dt
3.38ddd
5.10d
5.21dt
J5,6 2, J5,F 52.5
3.46ddd
J6,F 29
5.09s
5.32dt
3.45ddd
5.12s
J5,F 51
4.27br dd
J1,6 10.5, J6,F 27
5.63s
J1%,2%
4.11dt
J2%,3%a 8.5, J2%,3%b 8.5
4
J1%,2%
4.28dt
2%,3%a=J2%,3%b
1.92ddd
J3%a,3%b 13, J3%a,4%
2.02dt
4
4.08d
J1%,2% 0, J2%,3%a 5, J2%,3%b
1.68ddd
J3%a,3%b 13, J3%a,4% 9.5
2.11dd
4.34d
J2%,3%a 5
1.87ddd
J3%a,3%b 13, J3%a,4% 10
2.08dd
4.28d
0
J
8
H-3%a 1.82ddd
J3%a,3%b 12, J3%a,4%
H-3%b 2.36dt
1.91ddd
2.11dd
5
5
J2%,3%b=J3%b,4% 8.5
J3%b,4%
6
J3%b,4%
8
J3%b,4% 6
H-4%
4.65tt
J4%,5%a 5, J4%,5%b 8.5
4.43–4.52m
4.30m
4.39m
1.72m
4.42ddt
J4%,5%a 6, J4%,5%b
1.76m
8
H-5%a 1.65m
1.80dt
1.63m
J4%,5%a=J5%a,6% 6, J5%a,5%b 13.5
H-5%b 1.65m
H-6%a 3.27m
1.91–2.01m
3.39t
1.63m
2.59ddd
1.79m
2.67ddd
1.76m
2.70ddd
J5%a,6%=J5%b,6%
3.39t
5.74d
6
J5%a,6%a 7, J5%b,6%a 8, J6%a,6%b 12.5
H-6%b 3.27m
2.63ddd
4.97d
2.71ddd
5.03d
2.75ddd
5.03d
H-1¦
H-2¦
H-3¦
H-4¦
H-5¦
5.82d
J1¦,2¦
5.59dd
J2¦,3¦ 10
4.75dt
J3¦,4¦ 10, J3¦,NH-3¦ 8.5
4.02t
J4¦,5¦ 10
4.51dt
4
J1¦,2¦ 4
5.59dd
4.76dt
4.03t
3.39dd
J2¦,3¦ 10
2.98t
3.43dd
3.03t
3.46dd
3.04t
J3¦,4¦ 10
3.16t
3.33t
3.21t
J4¦,5¦ 10
3.75ddd
J5¦,6¦a 7, J5¦,6¦b
3.60dd
4.48dt
3.89t
3.72 ddd
J5¦,6¦a 4, J5¦,6¦b
3.79dd
3.82ddd
J5¦,6¦a 7, J5¦,6¦b 2
3.66dd
J5¦,6¦a 10, J5¦,6¦b
H-6¦a 3.81t
J6¦a,6¦b 10
H-6¦b 4.30dd
5
2
2
J6¦a,6¦b 12
3.80dd
4.46dd
3.76dd
3.86dd

332
Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
Table 3
13C chemical shifts (l, ppm) and coupling constants (JC,F, Hz) for 28a,b, 29a,b, 30a,b, 31, and 32b
Solvent C
Pyridine-d5
26% ND3 in D2O
28a
28b
30a
30b
31
29a
29b
32b
1
51.00d
J 4
51.47
51.31d
J 4
51.51d
J 4
49.35d
J 4
48.16d
J 4
48.24d
J 4
48.12d
J 3
2
3
35.13
51.45d
J 4
35.76
51.47
34.80
51.50d
J 5
77.18d
J 17
93.03d
J 183
81.35d
J 18
102.89
78.87
34.35
74.53
35.45
45.94
99.61
72.19
55.08
79.54
65.00
68.78
52.64
72.43
102.11
35.26
51.00d
J 4
74.56d
J 17
89.69d
J 182
80.94d
J 18
103.32
83.94
35.48
78.32
36.26
46.71
99.66
72.12
55.00
79.60
65.10
68.77
52.69
71.21
102.04
39.00
48.62d
J 3
79.86d
J 17
90.56d
J 179
86.18d
J 17
106.92
76.51
39.26
79.56
38.30
47.41
103.27
74.29
57.25
72.36
74.70
62.97
54.28
36.29
47.27d
J 3
79.20d
J 17
36.31
47.82d
J 4
83.14d
J 17
93.58d
J 178
84.37d
J 17
110.85
75.75
37.76
79.66
40.29
38.73
101.65
72.43
54.81
69.84
73.11
60.70
36.24
47.53d
J 2.5
79.34d
J 16
90.99d
J 176
84.97d
J 17
106.31
76.01
37.95
79.85
40.25
38.90
101.96
72.48
54.95
70.62
73.41
61.70
4
5
6
75.78d
J 17.5
90.68d
J 183
81.11d
J 17.5
99.21
78.61
34.28
74.47
35.63
48.37
99.67
72.16
54.99
79.56
65.11
68.79
78.80d
J 17
92.82d
J 183
81.15d
J 17.5
108.69
83.82
35.63
78.19
36.70
48.91
99.95
72.14
55.03
79.57
65.14
68.71
90.95d
J 177
84.90d
J 17.5
100.01
71.34
36.17
76.44
38.88
38.15
101.83
72.40
54.84
70.52
73.25
61.61
1%
2%
3%
4%
5%
6%
1¦
2¦
3¦
4¦
5¦
6¦
PhCH2N
PhCH2O
PhCH
72.05
102.00
71.25
102.17
matography on Kieselgel 60 (E. Merck), un-
less otherwise stated.
[10,11], [h]2D4 −33° (c 1.0, CHCl3), {lit. [24],
[h]2D2 −15.5° (c 2, CHCl3)}.
Computation.—All calculations were per-
formed on a Sun SPARC 2 Station with Ma-
teria Version 3.2 (Teijin System Technology,
Ltd., Hongou, Bunkyo-ku, Tokyo, Japan), us-
ing semiempirical MOPAC 93/PM3 [22] ac-
cording to the MOPAC 93 manual revision
Nr. 2 (Fujitsu Ltd., Nakase, Mihama-ku,
Chiba, Japan). Geometry optimization was
performed by the eigenvector following
method. The energy-minimal conformations
were searched initially by MM2UEC [23] ro-
tating the glycosidic bond in 15° steps, and the
conformations obtained were further opti-
mized with the MOPAC 93 with the COSMO
method (setting the dielectric constant as 78.3
to mimic the water effect).
1,2;5,6-Di-O-isopropylidene-3-O-(methyl-
thio)thiocarbonyl-h-
L
-glucofuranose
(1 ).—
L
1,2;5,6-Di-O-isopropylidene-a-
L
-glucofura-
nose [14,25] (7.70 g) was xanthated in THF
(154 mL) as described above to give a syrup
(10.44 g, quant.) [h]2D5 +33° (c 1.0, CHCl3).
1,2-O-Isopropylidene-3-O-(methylthio)thio-
carbonyl-h-
D
-glucofuranose
(2 ).—Com-
D
pound 1
D
(2.44 g) was hydrolyzed as reported
[10,11], [h]2D3 +40° (c 1.0, CHCl3) {lit. [24],
[h]2D4 −27.8° (c 2, 0.025 M HCl in 1:1 water–
EtOH)}.
1,2-O-Isopropylidene-3-O-(methylthio)thio-
carbonyl-h-
L
-glucofuranose (2 ).—Compound
L
1 (1.38 g) was treated with 0.4 M H2SO4 in
L
7:1 MeOH–water (28 mL) as described for 2
D
1,2;5,6-Di-O-isopropylidene-3-O-(methyl-
to give a solid (1.19 g, 97%), [h]2D3 −40° (c 1.0,
CHCl3).
thio)thiocarbonyl-h-
D
-glucofuranose
(1 ).—
D
Prepared according to the procedure reported
5,6-Di-O-acetyl-1,2-O-isopropylidene-3-O-

Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
333
made the decomposition of the reagent incom-
plete in the next water-treatment giving im-
(methylthio)thiocarbonyl - h -
D
- glucofuranose-
(3 ).—Compound 2 (1.82 g) was acetylated
D
D
pure 7 ; addition of 1,4-dioxane corrected the
D
as described for lit. [10,11] to give a syrup
(99%), [h]2D3 +11.5° (c 1.0, CHCl3) (lit. [11],
no data reported).
situation.) The organic solution was washed
with 5% aq KHSO4, dried (Na2SO4), and con-
centrated to give 7
D
as a relatively unstable
5,6-Di-O-acetyl-1,2-O-isopropylidene-3-O-
1
solid (6.70 g); H NMR (CDCl3): l 1.31, 1.50
[s of 3 H each, C(CH3)2], 1.79 (ddd, 1 H, J2,3a
4.5, J3a,3b 13.5, J3a,4 11 Hz, H-3a), 2.26 (dd, 1
H, J3b,4 4.5 Hz, H-3b), 2.40 (s, 3 H, Ts-CH3),
4.37 (dd, 1 H, H-6a), 4.52 (dd, 1 H, H-6b),
4.53 (m, 1 H, H-4), 4.74 (t, 1 H, H-2), 5.77 (d,
1 H, J1,2 4 Hz, H-1), 5.82 (dt, 1 H, H-5), 7.03,
7.51, and 8.18 (m of 1 H each, 3 imidazolyl-
H), 7.25 and 7.72 (ABq, 4 H, Ph-H of Ts).
3,5 - Dideoxy - 1,2 - O - isopropylidene - 6 - O-
(methylthio)thiocarbonyl - h -
(3
).—Syrup (96%), [h]2D3 −11° (c 1.0,
CHCl3).
5,6-Di-O-acetyl-3-deoxy-1,2-O-isopropyli-
dene-h- -ribo-hexofuranose (4 ).—Prepared
from 3 according to the method reported
L
- glucofuranose
L
D
D
D
[10,11], crystals (84%), mp 57–58 °C, [h]D24
+
19° (c 1.0, CHCl3) {lit. [26], [h]2D0 +20° (c 0.9,
CHCl3)}.
5,6-Di-O-acetyl-3-deoxy-1,2-O-isopropyli-
tosyl-h-
solution of 7
D
-erythro-hexofuranose (8
D
).—To a
dene-h-
L
-ribo-hexofuranose (4
L
).—Prepared
form 3
L
as described above, syrup (93%), [h]D23
D
(6.60 g) in toluene (268 mL)
were added Bu3SnH (11.5 mL, 43 mmol) and
AIBN (235 mg, 1.4 mmol), and the solution
was kept under Ar for 30 min at 80 °C. Con-
centration, followed by chromatography (2:1
−16° (c 1.4, CHCl3).
3 - Deoxy - 1,2 - O - isopropylidene - h -
D
- ribo-
by
hexofuranose (5
D).—Prepared from 4
D
Zemple´n deacetylation, crystals (quant.), mp
76–78 °C, [h]2D1 −18° (c 1.0, CHCl3) {lit. [27],
mp 82 °C, [h]2D1 −19.0° (c 1.7, CHCl3)}.
n-hexane–EtOAc) of the residue gave 8 as a
D
syrup (3.74 g, 78% based on 6
), [h]2D2 −8° (c
D
1.0, CHCl3), (lit. [12], [h]2D0 −2.9° (c 1.1,
3-Deoxy-1,2-O-isopropylidene-h-
ofuranose (5 ).—Prepared from 4
L
-ribo-hex-
1
MeOH); H NMR (500 MHz, CDCl3): l 1.29
L
L
(3.96 g) by
and 1.47 [s of 3 H each, C(CH3)2], 1.46 (ddd,
1 H, H-3a), 1.94 (m, 2 H, H-5), 2.09 (dd, 1 H,
H-3b), 2.45 (s, 3 H, Ts-CH3), 4.11 (ddd, 1 H,
J5a(5b),6a 6.5 and 8.0, J6a,6b 10.5 Hz, H-6a), 4.17
(ddd, 1 H, J5a(5b),6b 6.0 and 7.5 Hz, H-6b), 4.20
(m, 1 H, H-4), 4.68 (t, 1 H, H-2), 5.73 (d, 1 H,
H-1), 7.35 and 7.77 (ABq, 4 H, Ph-H of Ts).
Anal. Calcd for C16H22O6S: C, 56.12; H, 6.48;
S, 9.36. Found: C, 56.02; H, 6.52; S, 9.38.
3,5 - Dideoxy - 1,2 - O - isopropylidene - 6 - O-
methanolic NH3 to give crystals (2.70 g, 96%),
mp 78–80 °C, [h]2D3 +19° (c 1.0, CHCl3).
3-Deoxy-1,2-O-isopropylidene-6-O-tosyl-h-
D
-ribo-hexofuranose (6
D
).—Prepared from 5
D
by selective tosylation (TsCl–pyridine) [10,11]
as a syrup (88%), [h]2D6 −4° (c 1.0, CHCl3) (lit.
[11], no data reported).
3-Deoxy-1,2-O-isopropylidene-6-O-tosyl-h-
L
-ribo-hexofuranose (6
L
).—Prepared from 5
L
as described above, syrup (85%), [h]2D3 +5° (c
1.0, CHCl3).
tosyl-h-
ture of 6
L
-erythro-hexofuranose (8
L
).—A mix-
3-Deoxy-1,2-O-isopropylidene-5-O-(thiocar-
L
(4.53 g, 12.6 mmol), N,N%-thio-
bonylimidazolyl) - 6 - O - tosyl - h -
furanose (7 ).—A mixture of 6
D
- ribo - hexo-
(5.14 g, 14.3
carbonyldiimidazole (2.99 g, 15 mmol) and
imidazole (1.03 g, 15 mmol) in toluene (90
mL) was kept under Ar for 30 min at 70 °C.
AIBN (207 mg, 1.26 mmol) and Bu3SnH (16.9
mL, 63 mmol) were added with toluene (180
mL), and the mixture was kept under Ar for 1
h at 80 °C. Concentration of the solution,
followed by chromatography (3:1“1:1 n-hex-
D
D
mmol), N,N%-thiocarbonyldiimidazole (5.10 g,
25.7 mmol), and imidazole (1.95 g, 28.6 mmol)
in (CH2Cl)2 (100 mL) was kept for 30 min at
70 °C. Evaporation of the solvent together
with 1,4-dioxane gave a residue, which after
agitation in water (1 h), was extracted with
CH2Cl2. (If 1,4-dioxane was omitted, the
residue included a slight amount of thiocar-
bonyldiimidazole in the (CH2Cl)2 film adhered
to the surface of the solid particle, and this
ane–EtOAc) of the residue gave 8 as a syrup
L
(3.66 g, 85%), [h]S23 +9° (c 1.0, CHCl3); Anal.
Calcd for C16H22O6S: C, 56.12; H, 6.48; S,
9.36. Found: C, 55.89; H, 6.26; S, 9.08.

334
Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
Compound 11b: [h]2D3 −25° (c 1.2, CHCl3);
6-Azido-1,2-O-isopropylidene-3,5,6-trideoxy-
-erythro-hexofuranose (9 ).—A mixture
of 8 (155 mg, 0.45 mmol) and NaN3 (294
mg, 4.5 mmol) in DMF (3.1 mL) was stirred
for 30 min at 70 °C. Conventional work-up
gave, after chromatography (4:1 n-hexane–
h-D
D
1H NMR* (CDCl3): l 3.33 (s, 3 H, OCH3),
3.99 (br d, 1 H, J2,3a 5 Hz, H-2), 4.92 (s, 1 H,
H-1). Anal. Calcd for C14H19N3O3: C, 60.63;
H, 6.91; N, 15.15. Found: C, 60.67; H, 6.86;
N, 15.05.
D
EtOAC), gave 9
D
as a syrup (75.3 mg, 78%),
6-Azido-2-O-benzyl-3,5,6-trideoxy- -ery-
D
[h]2D4 −10° (c 1.1, CHCl3); IR (KBr): 2100
cm−1. Anal. Calcd for C9H15N3O3: C, 50.69;
H, 7.09; N, 19.71. Found: C, 50.96; H, 7.14;
N, 19.41.
thro-hexofuranose (12).—An anomeric mix-
ture of 11 (514 mg, 1.85 mmol) in 1:11 aq 12
M HCl–aq 80% AcOH (10 mL) was kept for
6 h at rt. Concentration with occasional addi-
tions of toluene and water gave a residue,
which was chromatographed (4:1 n-hexane–
EtOAc) to give 12 as a syrup (353 mg, 72%)
6-Azido-1,2-O-isopropylidene-3,5,6-trideoxy-
h-
8
L
-erythro-hexofuranose (9
(441 mg) in DMF (8.8 mL) was treated
L
).—Compound
L
1
similarly as above to give 9 as a syrup (229
L
together with 11 recovered (94 mg, 18%). H
mg, 83%), [h]2D3 +10° (c 1.0, CHCl3). Anal.
Calcd for C9H15N3O3: C, 50.69; H, 7.09; N,
19.71. Found: C, 50.93; H, 7.05; N, 19.47.
NMR (CDCl3): l 1.65–1.95 (m, 3 H, H-
3a,5a,5b), 2.16 (ddd, 1 H, H-3b, J2,3b 1, J3a,3b
13.5, J3b,4 6 Hz), 3.4 (m, 2 H, H-6a,6b), 4.13
(m, 1 H, H-2), 4.34 (m, 1 H, H-4), 4.55 [s,
C6H5CH2 (b)], 4.61 [d, J 4 Hz, C6H5CH2 (a)],
5.36 [d, 0.18 H, J 4 Hz, H-1(a)], 5.40 [d, 0.82
H, J 3 Hz, H-1(b)]. Anal. Calcd for
C13H17N3O3: C, 59.30; H, 6.51; N, 15.95.
Found: C, 59.12; H, 6.53; N, 15.74.
Methyl
hexofuranoside (10).—Methanolysis (1.5 h at
room temperature (rt)) of 9 (1.23 g, 5.76
6-azido-3,5,6-trideoxy-
D
-erythro-
D
mmol) in 1:5 aq 12 M HCl–MeOH (25 mL),
followed by neutralization (NaHCO3) and
subsequent chromatography (1:1 n-hexane–
EtOAc) gave 10 as a syrup (813 mg, 75%),
6-Azido-2-O-benzyl-3,5,6-trideoxy- -ery-
D
TLC (2:1 n-hexane–EtOAc): Rf 0.17 (a
thro-hexofuranosyl chloride (13).—A mixture
of 12 (251 mg, 0.95 mmol) and SOCl2 (2.5
mL) was kept for 3.5 h at rt. Concentration of
the solution gave 13 as a syrup (292 mg,
quant), which was used without purification;
1H NMR (CDCl3): l 4.22 [dt, 0.1 H, J1,2 3.5,
1
anomer), 0.2 (b anomer) (cf. 9
D
: Rf 0.7); H
NMR* (CDCl3): l 2.21 (br s, 1 H, OH-2),
3.34 (s, 2.6 H, b-OCH3), 3.49 (s, 0.4 H, a-
OCH3), 4.24 (br s, 1 H, H-2), 4.78 [s, 0.86 H,
H-1(b)], 4.87 [d, 0.14 H, J1,2 4.5 Hz, H-1(a)].
Anal. Calcd for C7H13N3O3: C, 44.91; H, 7.00;
N, 22.45. Found: C, 44.44; H, 7.17; N, 22.22.
Methyl 6-azido-2-O-benzyl-3,5,6-trideoxy-
J
2,3a=J2,3b 9 Hz, H-2(a)], 4.38 [dd, 0.9 H,
J2,3a(3b) 0.5 and 4 Hz, H-2(b)], 6.17 [s, 0.9 H,
H-1(b)], 6.25 [d, 0.1 H, H-1(a)].
D-erythro-hexofuranoside (11).—An anomeric
6-Azido-2-O-benzyl-3,5,6-trideoxy- -ery-
D
mixture of 10 (701 mg, 3.74 mmol) and NaH
(60% in mineral oil, 450 mg, 11.2 mmol) in
DMF (7.0 mL) was vigorously stirred for 5
min at 0 °C, BnBr (0.67 mL, 5.6 mmol) was
added, and the mixture was stirred for a fur-
ther 45 min. After addition of water, the
product was extracted with benzene and sub-
jected to chromatography (5:1 n-hexane–
EtOAc) to give 11a (133 mg, 13%) and 11b
(868 mg, 84%) as syrups.
thro-hexofuranosyl fluoride (14).—To an ice-
cooled solution of 12 (63.4 mg, 0.24 mmol) in
CH2Cl2 (1.3 mL) was added DAST (64 mL,
0.48 mmol) and the solution was kept for 20
min at that temperature. After addition of
excess CH2Cl2 and aq NaHCO3 (satd, 1 mL),
followed by stirring for 30 min, the mixture
was washed with water, dried (Na2SO4), and
concentrated to give, after chromatography
(7:1 n-hexane–EtOAc), 14a (12.2 mg, 19%)
and 14b (42.3 mg, 66%) as syrups.
Compound 11a: [h]2D1 +100° (c 1.0,
1
CHCl3); H NMR* (CDCl3): l 3.41 (s, 3 H,
Compound 14a: TLC (2:1 n-hexane–
OCH3), 3.98 (dt, 1 H, J1,2 4, J2,3a =J2,3b 8.5
Hz, H-2), 4.81 (d, 1 H, H-1). Anal. Calcd for
C14H19N3O3: C, 60.63; H, 6.91; N, 15.15.
Found: C, 60.64; H, 6.95; N, 15.18.
1
EtOAc): Rf 0.5; H NMR (CDCl3): l 1.76 (m,
2 H, H-5a,5b), 1.93 (ddd, 1 H, J2,3a 9, J3a,3b
12.5, J3a,4 4 Hz, H-3a), 2.24 (dt, 1 H, J2,3b
=

Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
335
1.60 (ddd, 1 H, J4,5b 4.5, J5a,5b 12.5, J5b,6a 9 Hz,
H-5b), 1.98 (dt, 1 H, J2,3b=J3b,4 8.5, J3a,3b 12
Hz, H-3b), 2.43 [s, 3 H, Ts(CH3)], 3.10 (ddd, 1
H, J5a,6a 6, J6a,6b 14 Hz, H-6a), 3.19 (ddd, 1 H,
J5a,6b and J5b,6b: 6.5 and 9 Hz, H-6b), 3.34 (s,
3 H, OCH3), 3.82 (dt, 1 H, J1,2 4, J2,3a 8.5 Hz,
H-2), 3.97 (tt, 1 H, H-4), 4.30 (ABq, 2 H,
C6H5CH2), 4.52 (s, 2 H, C6H5CH2), 4.69 (d, 1
H, H-1). Anal. Calcd for C28H33NO5S: C,
67.85; H, 6.71; N, 2.83; S, 6.47. Found: C,
67.59; H, 6.63; N, 2.74; S, 6.68.
J3b,4 9 Hz, H-3b), 3.38 (dt, 2 H, H-6a,6b), 4.05
(ddt, 1 H, J1,2 3, J2,F 19.5 Hz, H-2), 4.52 (m, 1
H, H-4), 4.62 (ABq, 2 H, PhCH2), 5.68 (dd, 1
H, J1,F 65 Hz, H-1); 19F NMR (CDCl3): l
−136.22 (dd, J1,F 65, J2,F 19.5 Hz, F-1).
Compound 14b: TLC (2:1 n-hexane–
1
EtOAc): Rf 0.55; H NMR (CDCl3): l 1.81
(dddd, 1 H, J2,3a 4.5, J3a,3b 13.5, J3a,4 3, J3a,F
Hz, H-3a), 1.86 (q, 2 H, J4,5=J5,6a=J5,6b
9
7
Hz, H-5a,5b), 2.20 (dd, 1 H, J3b,4 6.5 Hz,
H-3b), 3.44 (m, 2 H, H-6a,6b), 4.15 (dd, 1 H,
J2,F 3.5 Hz, H-2), 4.51 (m, 1 H, H-4), 4.56 (s,
2 H, PhCH2), 5.73 (d, 1 H, J1,F 64 Hz, H-1);
19F NMR (CDCl3): l −114.31 (ddd, J1,F 64,
J2,F 3.5, J3a,F 9 Hz, F-1).
b Anomer: TLC (2:1 n-hexane–EtOAc): Rf
1
0.55, [h]2D4 +17° (c 1.0, CHCl3); H NMR
(500 MHz, CDCl3): l 1.55–1.68 (m, 3 H,
H-3a,5a,5b), 1.96 (ddd, 1 H, J2,3b 0.5, J3a,3b 13,
J3b,4 6 Hz, H-3b), 2.42 [s, 3 H, Ts(CH3)],
3.13–3.19 (m, 1 H, H-6a), 3.16 (s, 3 H,
OCH3), 3.29 (ddd, 1 H, J5a,6b and J5b,6b: 5.5
and 10.5, J6a,6b 14 Hz, H-6b), 3.88 (br d, 1 H,
J2,3a 4.5 Hz, H-2), 4.10 (m, 1 H, H-4), 4.32
(ABq, 2 H, C6H5CH2), 4.48 (s, 2 H,
C6H5CH2), 4.79 (s, 1 H, H-1). Anal. Calcd for
C28H33NO5S: C, 67.85; H, 6.71; N, 2.83; S,
6.47. Found: C, 67.93; H, 6.78; N, 2.88; S,
6.41.
1,2 - O - Isopropylidene - 6 - tosylamido - 3,5,6-
trideoxy-h-
mixture of 9
L
-erythro-hexofuranose (15).—A
(385 mg, 1.80 mmol) and Raney
L
Ni in 2:1 THF–water (12 mL) was stirred
under H2 for 1 h at rt. Filtration through a
bed of Celite, followed by concentration gave
a residue, which was treated with TsCl (412
mg, 2.2 mmol) and Na2CO3 (230 mg, 2.2
mmol) in 2:1 CH2Cl2–water (10 mL) under
vigorous stirring for 1 h. The product was
chromatographed (3:2 n-hexane–EtOAc) to
give 15 as a syrup (532 mg, 87%), [h]2D3 +8° (c
2-O-Benzyl-6-N-benzyl-6-tosylamido-3,5,6-
trideoxy- -erythro-hexofuranose (18).—Hy-
L
1
1.1, CHCl3); H NMR* (CDCl3): l 1.30, 1.46
drolysis of 17 (1.75 g) as described for 12,
[s of 3 H each, C(CH3)2], 2.42 [s, 3 H,
Ts(CH3)], 5.20 (br dd, 1 H, TsNH-6), 5.76 (d,
1 H, J1,2 4 Hz, H-1). Anal. Calcd for
C16H23NO5S: C, 56.29; H, 6.79; N, 4.10; S,
9.39. Found: C, 56.03; H, 6.96; N, 4.27; S,
9.24.
followed by chromatography (20:1 CHCl3–
1
MeOH) gave 18 as a syrup (1.47 g, 86%). H
NMR (CDCl3): l 1.41–1.75 (m, 3 H, H-
3a,5a,5b), 1.96 (m, 1 H, H-3b), 2.42 [s, 3 H,
Ts(CH3)], 2.60 (br s, 0.6 H, OH-1), 3.06–3.37
(m, 2 H, H-6a,6b), 3.86–4.15 (m, 2 H, H-2,4),
4.29, 4.54 (ABq, 4 H, C6H5CH2), 4.48 (s, 2 H,
C6H5CH2), 5.27 (m, 1 H, H-1). Anal. Calcd
for C27H31NO5S·0.7H2O: C, 65.61; H, 6.61; N,
2.83. Found: C, 65.42; H, 6.28; N, 2.80.
Methyl 6-tosylamido-3,5,6-trideoxy- -ery-
L
thro-hexofuranoside (16).—Compound 15
(1.60 g, 4.68 mmol) was methanolyzed in 1:5
aq 12 M HCl–MeOH (32 mL) for 1 h at rt
and the product was chromatographed (30:1
CHCl3–MeOH) to give an anomeric mixture
of 16 as a syrup (1.44 g, 97%).
2-O-Benzyl-6-N-benzyl-6-tosylamido-3,5,6-
trideoxy- -erythro-hexofuranosyl chloride (19).
L
—A mixture of 18 (484 mg, 1.01 mmol) and
SOCl2 (4.8 mL) was kept for 3 h at rt. Careful
concentration gave 19 as an unstable syrup
(528 mg, quant), which was used without
purification.
Methyl 2 - O - benzyl - 6 - N - benzyl - 6 - tosyl-
amido-3,5,6-trideoxy- -erythro-hexofuranoside
L
(17).—Compound 16 (1.10 g) was benzylated
as described for 11 to give 17 as a syrup (1.75
g, quant). Analytical samples of the a and b
anomers were obtained by chromatography
(4:1“2:1 n-hexane–EtOAc).
2%,3%,4%,2¦,4¦,6¦ - Hexa - O - acetyl - 1,3,6%,3¦-
tetra-N-tosylkanamycin A (21).—An ice-cold
mixture of 1,3,6%,3¦-tetra-N-tosylkanamycin A
(20) [15] (5.00 g, 4.54 mmol) and AcCl (2.26
mL, 31.8 mmol) in pyridine (100 mL) was
a Anomer: TLC (2:1 n-hexane–EtOAc): Rf
1
0.4, [h]2D4 −54° (c 1.0, CHCl3); H NMR (500
MHz, CDCl3): l 1.47–1.55 (m, 2 H, H-3a,5a),

336
Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
1 H, J3,NH-3 9 Hz, TsNH-3), 10.03 (br d, 1 H,
J3¦,NH-3¦ 7 Hz, TsNH-3¦). The H-1, 3, and 4, 6
signals were assigned by 1H–1H COSY,
HMQC, and HMBC spectra. F NMR (pyri-
dine-d5): l −213.39 (dt, J4,F=J6,F 26, J5,F
kept for 6 h at rt. After addition of water (1.2
mL), followed by standing for 1 h, the solu-
tion was concentrated to give a residue, which,
after dissolving in CHCl3, was washed with aq
NaHCO3 (satd), aq 5% KHSO4, and water,
dried (Na2SO4), and concentrated to give 21
as an amorphous solid (6.06 g, 99%). An
analytical sample was prepared by chromatog-
raphy (3:1 CHCl3–acetone), [h]2D4 +48° (c 1.0,
19
50.5
Hz,
F-5).
Anal.
Calcd
for
C58H71FN4O24S4·H2O: C, 50.72; H, 5.36; N,
4.08; S, 9.34. Found: C, 50.63; H, 5.33; N,
4.25; S, 9.17.
1
5-Deoxy-5-epi-5-fluoro-1,3,6%,3¦-tetra-N-to-
sylkanamycin A (23).—A solution of 22
(monohydrate, 332 mg, 0.25 mmol) in 0.5%
NaOMe in MeOH (6.6 mL) was kept for 30
min at rt. Conventional purification gave 23
as an amorphous solid (268 g, 99%), TLC
(2:1:1 CHCl3–MeOH–aq 28% NH3): Rf 0.1;
CHCl3); H NMR* (pyridine-d5): l 1.90, 1.91,
2.04, 2.07, 2.08, 2.17, 2.17, 2.30, 2.31, 2.37 [s
of 3 H each, 4 Ts(CH3) and 6 Ac], 3.6–4.0 (m,
7 H, H-1,3,4,5,6,6%a,6%b), 4.71 (q, 1 H, J2¦,3¦
=
J3¦,4¦=J3¦,NH-3¦ 10 Hz, H-3¦), 6.01 (d, 1 H, J1%,2%
4 Hz, H-1%), 6.03 (d, 1 H, J1¦,2¦ 4 Hz, H-1¦),
6.61 (d, 1 H, OH-5); 8.52, 9.01 (d of 1 H each,
TsNH-1,3), 8.72 (m, 1 H, TsNH-6%), 10.08 (d,
1 H, J 7 Hz, TsNH-3¦). Anal. Calcd for
C58H72N4O25S4: C, 51.47; H, 5.36; N, 4.14; S,
9.47. Found: C, 51.17; H, 5.59; N, 4.39; S,
9.09.
1
[h]2D4 +50° (c 1.0, DMF); H NMR* (500
MHz, pyridine-d5): l 2.09, 2.15, 2.21, 2.26 [s
of 3 H each, 4 Ts(CH3)], 3.71 (dt, 1 H, H-1),
3.82 (m, 2 H, H-6%a,6%b), 4.02 (dd, 1 H, J1,6 11,
J6,F 26.5 Hz, H-6), 4.08 (m, 1 H, H-3), 4.19
(dd, 1 H, J3,4 11, J4,F 26.5 Hz, H-4), 4.46 (t, 1
H, H-3¦), 5.45 (d, 1 H, H-1¦), 5.48 (d, 1 H,
H-1%), 5.99 (d, 1 H, J5,F 51.5 Hz, H-5), 8.44 (br
2%,3%,4%,2¦,4¦,6¦-Hexa-O-acetyl-5-deoxy-5-
epi-5-fluoro-1,3,6%,3¦-tetra-N-tosylkanamycin
A (22).—To a cold (−20 °C) solution of 21
(5.91 g, 4.37 mmol) in CH2Cl2 (120 mL),
DAST (2.89 mL, 22 mmol) was added, and
the solution was kept for 2.5 h at 0 °C.
Aqueous NaHCO3 (satd, 400 mL) was added
under vigorous stirring, and the product was
extracted with CHCl3 to give 22 as an amor-
phous solid (6.93 g, quant). An analytical
sample was prepared by chromatography
(20:1 CHCl3–MeOH); HPTLC (20:1 CHCl3–
MeOH), Rf 0.6 (cf. 21: Rf 0.52), [h]2D3 +56.5°
s, 1 H, TsNH-1), 8.61 (br t, 1 H, J6%a(6%b),NH-6%
6
Hz, TsNH-6%), 8.96 (br d, 1 H, J3,NH-3 7 Hz,
TsNH-3), 9.41 (br s, 1 H, TsNH-3¦); 19F
NMR (pyridine-d5): l −212.05 (dt, J4,F=J6,F
27.5, J5,F 51.5 Hz, F-5); 13C NMR (pyridine-
d5): l 21.14, 21.29, 21.35, 21.45 [4 Ts(CH3)],
34.68 (C-2), 45.08 (C-6%), 50.94 (d, J 4 Hz,
C-3), 51.80 (d, J 4 Hz, C-1), 61.67 (C-3¦),
62.11 (C-6¦), 69.99 (C-4¦), 72.25 (C-2¦), 72.39
(C-4%,5%), 73.47 (C-2%), 74.60 (C-3%), 75.43 (C-
5¦), 77.40 (d, J 17 Hz, C-4), 82.58 (d, J 17 Hz,
C-6), 90.82 (d, J 182 Hz, C-5), 98.67 (C-1%),
103.66 (C-1¦). The H-1, 3, 4, 6 signals were
assigned by 1H–1H COSY, HMQC, and
HMBC spectra. Anal. Calcd for C46H59FN4O-
18S4: C, 50.08; H, 5.39; N, 5.08; S, 11.62.
Found: C, 50.04; H, 5.56; N, 5.02; S, 11.32.
1
(c 1.0, CHCl3); H NMR* (500 MHz, pyri-
dine-d5): l 1.96, 1.99, 2.017, 2.024, 2.04, 2.16,
2.17, 2.20, 2.31, 2.35 [s of 3 H each, 4 Ts(CH3)
and 6 Ac], 3.62 (ddd, 1 H, J5%,6%a 2.5, J6%a,6%b
13.5, J6%a,NH-6% 5 Hz, H-6%a), 3.70 (ddd, 1 H,
J5%,6%b 3.5, J6%b,NH-6% 8 Hz, H-6%b), 3.85 (ddt, 1 H,
J1,2ax =J1,6 10.5, J1,2eq 4.5, J1,NH-1 7 Hz, H-1),
4.04 (m, 1 H, H-3), 4.13 (dd, 1 H, J6,F 27.5 Hz,
H-6), 4.26 (dd, 1 H, J3,4 11, J4,F 27.5 Hz, H-4),
4.55 (br d, 1 H, J6¦a,6¦b 13.5 Hz, H-6¦a), 4.64
(dd, 1 H, J5¦,6¦b 4 Hz, H-6¦b), 4.75 (m, 1 H,
H-3¦), 4.90 (dd, 1 H, J1%,2% 4, J2%,3% 10.5 Hz,
H-2%), 5.45 (dd, 1 H, J1¦,2¦ 4, J2¦,3¦ 10.5 Hz,
H-2¦), 5.46 (br d, 1 H, J5,F 51.5 Hz, H-5), 5.59
(d, 1 H, H-1¦), 5.85 (d, 1 H, H-1%), 7.86 (d, 1
H, TsNH-1), 8.69 (dd, 1 H, TsNH-6%), 9.30 (d,
6-O-(3-Deoxy-3-tosylamido-h- -glucopy-
D
ranosyl)-2,5-dideoxy-5-epi-5-fluoro-1,3-di-
N-tosylstreptamine (24).—To a solution of 23
(1.00 g, 0.91 mmol) in MeOH (100 mL) was
added NaIO4 (1.94 g, 9.07 mmol), and the
mixture was stirred for 2 h at rt. Ethylene
glycol (0.5 mL) was added, and after stirring
for 30 min, the solution was concentrated. The
residue was shaken with water and the insolu-

Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
337
deoxy-3-tosylamido-h- -glucopyranosyl)-4-O-
D
ble matter was collected, and dried. To a
MeOH solution (20 mL) of the solid was
added NaBH4 (858 mg, 22.7 mmol), and the
mixture was kept for 2.5 h at rt. After acetone
(6.67 mL) had been added, followed by stir-
ring (30 min), the mixture was neutralized
with aq 1 M HCl. After concentration, the
residue dissolved in 1.5 M HCl in MeOH (20
mL) was kept for 5 h at rt. Concentration of
the solution gave a residue, which was thor-
oughly washed with water and dried to give 24
as a practically pure solid (639 mg, 87%), TLC
(1.5:1:1 CHCl3–MeOH–aq 28% NH3): Rf
0.33 (cf. 23: Rf 0.2), [h]1D9 +25° (c 1.0, DMF);
1H NMR* (pyridine-d5): l 2.08, 2.18, 2.22 [s
acetyl-2,5-dideoxy-5-epi-5-fluoro-1,3-di-N-to-
sylstreptamine (27).—To a solution of 25 (691
mg, 0.79 mmol) in 9:1 dry Me2SO–pyridine
[20] (6.9 mL) was added N-acetylimidazole
(174 mg, 1.58 mmol), and the solution was
kept for 65 h at rt. Water was added, and
after stirring for 30 min, the precipitate ob-
tained was filtered, dried, and chro-
matographed (15:1 CHCl3–MeOH) to give 26
(342 mg, 47%) and 27 (380 mg, 49%) as
amorphous solids. Compound 27 could be
converted into 25 by deacetylation, as de-
scribed for 23.
Compound 26: TLC (7:1 CHCl3–MeOH):
Rf 0.45 (cf. 25: Rf 0.25); [h]2D3 −14° (c 1.0,
of 3 H each, 3 Ts(CH3)], 5.57 (d, 1 H, J1%,2%
4
1
CHCl3); H NMR* (pyridine-d5): l 2.35 (s, 3
Hz, H-1%), 5.71 (br d, 1 H, J5,F 52 Hz, H-5),
8.45 (br s, 1 H, TsNH-1 or 3), 9.27 (br d, 1 H,
H, AcO-2%), 5.56 (s, 1 H, C6H5CH), 5.58 (dd,
1 H, J1%,2% 4, J2%,3% 11 Hz, H-2%), 5.59 (dt, 1 H,
J4,5=J5,6 1.5, J5,F 52 Hz, H-5), 5.83 (d, 1 H,
19
TsNH-3 or 1), 9.43 (br d, 1 H, TsNH-3%); F
NMR (pyridine-d5): l −211.66 (dt, J4,F=J6,F
30, J5,F 52 Hz, F-5). Anal. Calcd for
C33H42FN3O12S3·H2O: C, 49.18; H, 5.50; N,
5.21. Found: C, 48.96; H, 5.43; N, 5.10.
6-O-(4,6-O-Benzylidene-3-deoxy-3-tosyl-
19
H-1%); F NMR (pyridine-d5): l −211.93 (dt,
J
4,F=J6,F 29, J5,F 52 Hz, F-5). Anal. Calcd for
C42H48FN3O13S3: C, 54.95; H, 5.27; N, 4.58; S,
10.48. Found: C, 54.62; H, 5.47; N, 4.71; S,
10.74.
amido-h- -glucopyranosyl)-2,5-dideoxy-5-epi-
D
Compound 27: TLC (7:1 CHCl3–MeOH):
5-fluoro-1,3-di-N-tosylstreptamine (25).—A
mixture of 24 (986 mg, 1.25 mmol),
C6H5CH(OMe)2 (0.38 mL, 2.50 mmol), and
anhyd p-toluenesulfonic acid (43 mg) in DMF
(10 mL) was stirred for 1 h under vacuum (35
mmHg) at 60 °C, whereupon removal of the
MeOH liberated occurred. After pouring the
solution into aq NaHCO3 (satd), the precipi-
tate was washed thoroughly with water, dried,
and chromatographed (10:1 CHCl3–MeOH)
to give 25 as an amorphous solid (1.08 g,
Rf 0.65; [h]2D2 +10° (c 1, CHCl3); H NMR*
1
(pyridine-d5): l 1.83 (s, 3 H, AcO-4), 2.33 (s, 3
H, AcO-2%), 4.07 (m, 1 H, H-1), 4.20 (m, 1 H,
H-3), 5.45 (ddd, 1 H, J3,4 11, J4,5 2, J4,F 28 Hz,
H-4), 5.57 (s, 1 H, C6H5CH), 5.58 (dd, 1 H,
J1%,2% 4, J2%,3% 10.5 Hz, H-2%), 5.62 (br d, 1 H, J5,F
52 Hz, H-5), 5.81 (d, 1 H, H-1%); 19F NMR
(pyridine-d5): l −211.50 (dt, J4,F =J6,F 28.5,
J5,F 52.5 Hz, F-5). Anal. Calcd for
C44H50FN3O14S3·H2O: C, 54.03; H, 5.63; N,
4.30; S, 9.83. Found: C, 54.33; H, 5.31; N,
4.33; S, 9.72.
1
quant), [h]2D1 +8° (c 1.0, CHCl3); H NMR
(pyridine-d5): l 2.07, 2.14, 2.22 [s of 3 H each,
3 Ts(CH3)], 5.44 (s, 1 H, C6H5CH), 5.54 (br.
d, 1 H, J5,F 53 Hz, H-5), 5.57 (d, 1 H, J1%,2% 3.5
Hz, H-1%), 6.8–8.4 [17 H, 3 Ts(C6H4),
C6H5CH], 8.39 (s), 9.35 (d), 9.57 (d) (1 H
6-O-(2-O-Acetyl-4,6-O-benzylidene-3-deoxy-
3 - tosylamido - h -
azido-2-O-benzyl-3,5,6-trideoxy-h-
-i- -erythro-hexofuranosyl)-2,5-dideoxy-5-
D
- glucopyranosyl) - 4 - O - (6-
D
- (28a) and
D
epi-5-fluoro-1,3-di-N-tosylstreptamine (28b).
—A mixture of 26 (430 mg, 0.47 mmol), 13
(110 mg, 0.39 mmol), and CaSO4 (Drierite,
530 mg), in THF (1.1 mL) was stirred for
30 min. Then Hg(CN)2 (197 mg, 0.78 mmol)
was added, and stirring was continued for
1 h at rt in the dark. After filtration through
a bed of Celite and CH2Cl2, the organic
solution was washed with aq NaHCO3 (satd)
and water, dried (Na2SO4), and concen-
19
each, TsNH-1,3,3%); F NMR (pyridine-d5): l
−211.18 (dt, J4,F=J6,F 27.5, J5,F 51 Hz, F-5).
Anal. Calcd for C40H46FN3O12S3: C, 54.84; H,
5.29; N, 4.80; S, 10.98. Found: C, 54.43; H,
5.41; N, 4.89; S, 10.96.
6-O-(2-O-Acetyl-4,6-O-benzylidene-3-deoxy-
3 - tosylamido - h - - glucopyranosyl) - 2,5 - dide-
D
oxy-5-epi-5-fluoro-1,3-di-N-tosylstreptamine
(26) and 6-O-(2-O-acetyl-4,6-O-benzylidene-3-

338
Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
debenzylidenation). After filtration, the sol-
vent was evaporated azeotropically with the
aid of toluene and water. To the residue in
liquid NH3 (ꢀ40 mL) at −50 °C, sodium
(ꢀ200 mg) was added, and after 1 h (detosy-
lation, debenzylation, and debenzylidenation
occurred during this operation) MeOH was
added until the blue color disappeared. Excess
NH3 was removed by gradually raising the
temperature to rt. The residue dissolved in
water, after neutralization with Dowex 50W-
X2 resin (H+ form), was chromatographed
with the same resin with aq 1 M NH4OH. The
ninhydrin-positive fractions were collected,
concentrated, and again chromatographed
trated. Chromatography (1:1“2:3 n-hexane–
EtOAc, changed gradually) of the residue
gave 28 as an anomeric mixture (a:b=1:1,
264 mg, 58% based on 13) together with 26
recovered (180 mg). Pure anomers were pre-
pared by repeated chromatographys (2:1“1:1
toluene–EtOAc).
Compound 28a: TLC (1:2 n-hexane–
EtOAc): Rf 0.35; [h]2D5 +46° (c 1.0, CHCl3);
1H NMR (500 MHz, pyridine-d5; see Table 2
for the signals not included below): l 2.14,
2.21, 2.24 [s of 3 H each, 3 Ts(CH3)], 2.33 (s,
3 H, Ac), 4.67 (ABq, 2 H, C6H5CH2), 5.57 (s,
1 H, C6H5CH), 8.09, 8.72 (d of 1 H each, J 6
Hz, TsNH-1,3), 9.83 (d, 1 H, J 8 Hz, TsNH-
3¦); 19F NMR (pyridine-d5): l −211.83 (dt,
with CM-Sephadex C-25 (0.05“0.2
M
J
4,F=J6,F 27.5, J5,F 52 Hz, F-5); 13C NMR
NH4OH, gradual change) to give 29a as a
solid (30.5 mg, 33% as 0.7H2CO3 salt; it is
generally difficult to obtain aminoglycoside
antibiotics as the free base due to their ab-
sorbing CO2 readily from the air during the
isolation process; the situation was the same
with 29b and 32b), [h]2D5 +154° (c 0.9, water);
19F NMR (26% ND3 in D2O): l −213.10 (dt,
(pyridine-d5; see Table 3 for other signals): l
21.08, 21.21, 21.25, 21.28 [3 Ts(CH3) and
CH3CO]. Anal. Calcd for C55H63FN6O15S3·
0.5H2O: C, 56.35; H, 5.50; N, 7.17; S, 8.20.
Found: C, 56.40; H, 5.74; N, 7.31; S, 8.19.
Compound 28b: TLC (1:2 n-hexane–
EtOAc): Rf 0.5; [h]2D5 −1.5° (c 1.0, CHCl3); 1H
NMR (500 MHz, pyridine-d5; see Table 2 for
the signals not included below): l 2.14, 2.20,
2.30 [s of 3 H each, 3 Ts(CH3)], 2.34 (s, 3 H,
Ac), 4.61 (s, 2 H, C6H5CH2), 5.63 (s, 1 H,
C6H5CH), 8.03 (d, 1 H, TsNH-1), 9.28 (d, 1
H, J3,NH-3 7 Hz, TsNH-3), 9.85 (d, 1 H, J 8
Hz, TsNH-3¦); 19F NMR (pyridine-d5): l −
J4,F=J6,F 29, J5,F 52.5 Hz, F-5). Anal. Calcd
for C18H35FN4O8·0.7H2CO3: C, 45.11; H, 7.37;
N, 11.25; F, 3.82. Found: C, 45.23; H, 7.65;
N, 11.04; F, 3.50.
4-O-(6-Amino-3,5,6-trideoxy-i-
hexofuranosyl)-6-O-(3-amino-3-deoxy-h-
D
-erythro-
D
-
glucopyranosyl) - 2,5 - dideoxy - 5 - epi - 5 - fluoro-
streptamine (29b).—A solution of 28b (697
mg, 0.60 mmol) was deacetylated (0.1%
NaOMe in 1:1 CHCl3–MeOH) and, after neu-
tralization [Dowex 50W-X2 resin (H+ form,
stored in MeOH)], was deprotected with
sodium (ꢀ670 mg) in 10:1 liquid NH3–THF
(ꢀ140 mL) as described for 29a to give 29b
as a solid (121 mg, 42% as 0.7H2CO3 salt),
[h]2D5 +46° (c 0.8, water); 19F NMR (26%
ND3 in D2O): l −212.68 (dt, J4,F=J6,F 29,
J5,F 52 Hz, F-5). Anal. Calcd for
C18H35FN4O8·0.7H2CO3: C, 45.11; H, 7.37; N,
11.25; F, 3.82. Found: C, 45.24; H, 7.65; N,
11.17; F, 3.58.
13
212.76 (dt, J4,F=J6,F 27, J5,F 51 Hz, F-5); C
NMR (pyridine-d5; see Table 3 for the signals
not included below): l 21.09, 21.23, 21.27
(two overlapped signals), [3 Ts(CH3) and
CH3CO]. Anal. Calcd for C55H63FN6O15S3: C,
56.79; H, 5.46; N, 7.22; S, 8.27. Found: C,
57.05; H, 5.49; N, 7.18; S, 8.27.
4-O-(6-amino-3,5,6-trideoxy-h-
D
-erythro-
hexofuranosyl)-6-O-(3-amino-3-deoxy-h-
D-
glucopyranosyl) - 2,5 - dideoxy - 5 - epi - 5 - fluoro-
streptamine (29a).—A solution of 28a (218
mg, 0.187 mmol) in 0.5% NaOMe in 9:1 pyri-
dine–MeOH (4.4 mL) was kept for 1 h at rt.
The mixture was poured into aq 5% KHSO4
(440 mL) under vigorous stirring, and the
precipitate was filtrated, washed with water,
and dried, which was hydrogenated with pal-
ladium black in 1:1 DMF–aq 80% AcOH (21
mL) for 4 h under hydrogen bubbling (reduc-
tion of N3 with partial debenzylation and
6-O-(2-O-Acetyl-4,6-O-benzylidene-3-deoxy-
3-tosylamido-h-
benzyl - 6 - N - benzyl - 6 - tosylamido - 3,5,6 - tri-
deoxy- -erythro-hexofuranosyl)-2,5-dideoxy-
5-epi-5-fluoro-1,3-di-N-tosylstreptamine (30).
D
-glucopyranosyl)-4-O-(2-O-
L

Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
339
H-4¦), 4.18–4.32 (m, 4 H, H-4,6,4%,6¦b), 4.41
(ABq, 2 H, C6H5CH2-6%), 4.47 (ABq, 2 H,
C6H5CH2-2%), 4.48–4.55 (m, 1 H, H-5¦), 4.77
(dt, 1 H, J2¦,3¦ 10.5, J3¦,NH-3¦ 9 Hz, H-3¦), 5.52
(s, 1 H, H-1%), 5.58 (dd, 1 H, J1¦,2¦ 4 Hz, H-2¦),
5.59 (s, 1 H, C6H5CH), 5.75 (br d, 1 H, J5,F 52
Hz, H-5), 5.81 (d, 1 H, H-1¦); 8.25, 8.31 (br d
of 1 H each, TsNH-1,3), 9.82 (d, 1 H, TsNH-
3¦); 19F NMR (pyridine-d5): l −212.56 (dt,
—A mixture of 26 (541 mg, 0.59 mmol), 19
,
(246 mg, 0.49 mmol), and 4 A molecular
sieves (246 mg) in CH2Cl2 (2.5 mL) was stirred
for 30 min at rt. Then Hg(CN)2 (248 mg, 0.98
mmol) and s-collidine (0.26 mL, 1.96 mmol)
were added, and the mixture was stirred for 16
h at rt in the dark. After filtration through a
bed of Celite and CH2Cl2, the solution was
washed with aq NaHCO3 (satd), aq 5%
KHSO4, and water, dried (Na2SO4), and con-
centrated. Chromatography (2:1 toluene–
EtOAc) of the residue gave 30b (b anomer,
153 mg) as a solid and a mixture of the
anomers (30a,b, 62 mg) (32% in total). An
analytical sample of 30a was obtained by re-
peated chromatographys (3:1“1:1 toluene–
EtOAc with gradual change).
J
4,F=J6,F 26, J5,F 52 Hz, F-5). Anal. Calcd for
C69H77FN4O17S4·H2O: C, 59.21; H, 5.69; N,
4.00. Found: C, 59.32; H, 5.60; N, 3.98.
4-O-(6-Amino-3,5,6-trideoxy-i-
hexofuranosyl)-6-O-(3-amino-3-deoxy-h-
L
-erythro-
D
-
glucopyranosyl) - 2,5 - dideoxy - 5 - epi - 5 - fluoro-
streptamine (32b).—Compound 30b (363 mg,
0.263 mmol) was deacetylated as described for
29a and the product was deprotected with
sodium in liquid ammonia as described for
29b to give the 6%-N-benzyl derivative 31 (152
30a: TLC (1:1 toluene–EtOAc): Rf 0.4, [h]D19
1
+1° (c 1.0, CHCl3); H NMR (500 MHz,
pyridine-d5): l 1.56–1.70 (m, 3 H, H-
3%a,5%a,5%b), 1.89 (q, 1 H, H-2ax), 2.10 (m, 1 H,
H-3%b), 2.14, 2.22, 2.26, 2.27 [s of 3 H each, 4
Ts(CH3)], 2.28 (s, 3 H, Ac), 2.82 (dt, 1 H,
H-2eq), 3.36 (m, 2 H, H-6%a,6%b), 3.82 (t, 1 H,
H-6¦a), 3.86 (m, 1 H, H-1), 3.93–4.02 (m, 3 H,
H-3,2%,4¦), 4.17–4.36 (m, 4 H, H-4,6,4%,6¦b),
4.41 (ABq, 2 H, C6H5CH2-6%), 4.45 (m, 1 H,
H-5¦), 4.87 (ABq, 2 H, C6H5CH2-2%), 4.74 (m,
1 H, H-3¦), 5.56 (dd, 1 H, H-2¦), 5.59 (s, 1 H,
C6H5CH), 5.60 (d, 1 H, J1%,2% 4 Hz, H-1%), 5.65
(br d, 1 H, J5,F 51.5 Hz, H-5), 5.76 (d, 1 H,
J1¦,2¦ 4 Hz, H-1¦), 8.00 (d, 1 H, J1,NH-1 6 Hz,
TsNH-1), 8.59 (d, 1 H, J3,NH-3 7.5 Hz, TsNH-
3), 9.82 (d, 1 H, J3¦,NH-3¦ 8.5 Hz, TsNH-3¦);
1
mg); H NMR* (500 MHz, pyridine-d5): l
1.90 (dt, 1 H, J 7, 7, 14 Hz, H-5%a), 2.03 (ddd,
1 H, J2%,3%a 4.5, J3%a,3%b 13, J3%a,4% 10 Hz, H-3%a),
2.11 (dt, 1 H, J 7, 7, 14 Hz, H-5%b), 2.24 (dd,
1 H, J3%b,4% 6 Hz, H-3%b), 2.84 (m, 2 H, H-
6%a,6%b), 3.81 (s, 2 H, C6H5CH2-6%), 4.69 (d, 1
H, H-2%), 4.81 (m, 1 H, H-4%), 5.46 (d, 1 H,
J
1¦,2¦ 4 Hz, H-1¦) 5.76 (s, 1 H, H-1%), 7.25–7.49
19
(5 H, C6H5CH2-6%); F NMR (pyridine-d5): l
−211.08 (dt, J4,F=J6,F 29, J5,F 53 Hz, F-5).
Compound 31 (142 mg) dissolved in 80% aq
AcOH (10 mL) was hydrogenated with palla-
dium black under bubbling of hydrogen as
described for 29a to give, after chromatogra-
phy (CM-Sephadex C-25, aq 0.05“0.2 M
NH4OH), 32b as a solid (20.4 mg, 15% based
on 30b as H2CO3 salt) with 31 recovered (80
mg), [h]1D9 +127° (c 0.5, water); 19F NMR
(26% ND3 in D2O): l −213.13 (dt, J4,F=J6,F
29, J5,F 52.5 Hz, F-5). Anal. Calcd for
C18H35FN4O.8H2CO3·H2O: C, 42.69; H, 7.35;
N, 10.48. Found: C, 42.61; H, 7.55; N, 10.54.
19F NMR (pyridine-d5): l −212.09 (dt, J4,F
=
J6,F 28, J5,F 51.5 Hz, F-5). Anal. Calcd for
C69H77FN4O17S4: C, 59.98; H, 5.62; N, 4.06.
Found: C, 59.88; H, 5.62; N, 3.90.
30b: TLC (1:1 toluene–EtOAc): Rf 0.55,
1
[h]1D9 +8° (c 1.0, CHCl3); H NMR (500
MHz, pyridine-d5): l 1.66 (ddd, 1 H, J2%,3%a 5,
J3%a,3%b 13, J3%a,4% 10 Hz, H-3%a), 1.84–2.00 (m, 3
H, H-2ax,3%b,5%a), 2.12 (m, 1 H, H-5%b), 2.14,
2.21, 2.24, 2.28 [s of 3 H each, 4 Ts(CH3)],
2.31 (s, 3 H, Ac), 2.72 (dt, 1 H, J1,2eq =J2eq,3
4.5, J2ax,2eq 13.5 Hz, H-2eq), 3.29 (ddd, 1 H,
J5%a(5%b),6%a 6.5 and 8.5, J6%a,6%b 14 Hz, H-6%a), 3.56
(ddd, 1 H, J5%a(5%b),6%b 5 and 9 Hz, H-6%b), 3.81
(t, 1 H, J5¦,6¦a=J6¦a,6¦b 10.5 Hz, H-6¦a), 3.96
(br d, 1 H, J2%,3%a 5 Hz, H-2%), 3.98–4.07 (m, 2
H, H-1,3), 4.03 (t, 1 H, J3¦,4¦=J4¦,5¦ 10.5 Hz,
Acknowledgements
We express deep thanks to Hiroko Hino of
Institute of Microbial Chemistry and Yoshiko
Koyama of our Institute for of elemental
analyses and NMR spectra, respectively.

340
Y. Kobayashi et al. / Carbohydrate Research 329 (2000) 325–340
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