Stereoselective synthesis and biological activity of novel spiro-oxazinanone-C-glycosides

Article abstract of DOI:10.1016/j.tet.2008.08.002

The stereoselective synthesis of novel spiro-oxazinanone nucleosides 9 and 10 has been achieved by microwave assisted 1,3-dipolar cycloaddition of exo-glucal (1) and nitrones (2), and followed by reduction, stereospecific recyclization, and catalytic depr

Full text of DOI:10.1016/j.tet.2008.08.002

Tetrahedron 64 (2008) 9911–9920
Contents lists available at ScienceDirect
Tetrahedron
journal homepage: www.elsevier.com/locate/tet
Stereoselective synthesis and biological activity of novel spiro-
oxazinanone-C-glycosides
,
b
,
a
a
a
a
a
a,b
Xiaoliu Li a , Rui Wang , Yanpo Wang , Hua Chen , Zhiwei Li , Cuilan Ba , Jinchao Zhang
*
a Laboratory of Chemical Biology, College of Chemistry and Environmental Science, Hebei University, Baoding, Hebei 071002, China
b Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, Hebei University, Baoding, Hebei 071002, China
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 18 April 2008
Received in revised form 29 July 2008
Accepted 5 August 2008
Available online 8 August 2008
The stereoselective synthesis of novel spiro-oxazinanone nucleosides 9 and 10 has been achieved by
microwave assisted 1,3-dipolar cycloaddition of exo-glucal (1) and nitrones (2), and followed by re-
duction, stereospecific recyclization, and catalytic deprotection. The structures of the spironucleosides
were determined according to the 1H NMR, 13C NMR, 2D NMR, MS, and X-ray analyses, and the biological
activities of the title compounds against glycosidases (
cytotoxicity were also evaluated.
a
-amylase,
a-glucosidase, and b-glucosidase) and
Keywords:
Ó 2008 Elsevier Ltd. All rights reserved.
Spironucleoside
Microwave irradiation
1,3-Dipolar cycloaddition
Nitrones
exo-Glycal
1. Introduction
type of spironucleosides with six-membered oxazinanone. From
this point of view, we would like to report herein a stereo-
selective synthesis of novel aryl substituted spiro-oxazinanone
glucosides 9 and 10 as the continuation of our work3o,7,8 on the
synthesis of functional C-glycosides applying the 1,3-dipolar cy-
cloaddition of exo-glucal (1) with nitrones (2) (see Scheme 1 and
Table 1).
Anomeric spironucleoside is a class of spiranic sugar de-
rivatives of which the anomeric carbon belongs simultaneously to
the sugar ring and to a heterocyclic base.1 The synthesis of the
spironucleoside derivatives has attracted considerable attention in
recent years1–4 due to their potential biological activities.5 How-
ever, most of such compounds focused on the analogues of nat-
ural spironucleoside (þ)-hydantocidin (A) in which the
heterocycle was a five-membered ring.3 The spironucleoside with
six-membered or seven-membered heterocycle has not been
investigated extensively,4 although the six-membered 1,3-oxa-
zinan-2-one ring (B) exists in many drug molecules as the core
substructure and its derivatives are of very important biological
activities.6
O
O
HO
HO
HO
R3
O
O
NH
O
O
RN
N
NHR2
OH
OH
H
R1
HO
HO
OH
A: (+)-Hydantocidin
B: 1,3-Oxazinan-2-one C: Amino-C-glycosides
2. Results and discussion
The six-membered ring of 1,3-oxazinan-2-one (B) can be
conventionally constructed by the cyclization of 1,3-amino alco-
hols. Recently, we have developed a convenient method to access
the amino-C-glycosides (C), a typical 1,3-amino alcohol with
a sugar moiety, by the 1,3-dipolar cycloaddition of exo-glycal with
nitrones, and then reductive cleavage of the O–N bond.7 We
envisaged that such amino-C-glycosides would be a very good
precursor for constructing oxazinanone derivatives containing
sugar moiety, which would provide a convenient access to a new
The synthetic pathway is shown in Scheme 1. Following the
microwave assisted procedure we recently developed,8 the cyclo-
addition of 1 and 2a–i was carried out very efficiently in diglyme
solution under microwave irradiation at 200 ꢀC for 2 min in
a sealed pressure vial and stereoselectively afforded the corre-
sponding adducts of spiro-isoxazolidine glycosides as the
a- and
b-anomeric mixture of 3a–i and 4a–i in 30–80% yield. The reductive
cleavage of O–N bond was completed by the treating the mixture
with Zn/AcOH/H2O to generate the intermediate of amino-C-gly-
coside derivatives 5 and 6, which were reacted with triphosgene9 to
form the cyclized key intermediate of spiro-oxazinanone
* Corresponding author. Tel./fax: þ86 312 5971116.
E-mail address: lixl@hbu.cn (X. Li).
0040-4020/$ – see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2008.08.002

9912
X. Li et al. / Tetrahedron 64 (2008) 9911–9920
Ar
BnO
BnO
BnO
Ar
BnO
BnO
O
N
O
O
O
i
BnO
+
Ar
N
+
O
N
OBn
4a~i
OBn
BnO
BnO
CH3
O
BnO
OBn
3a~i
2a~i
1
ii
Ar
BnO
BnO
NH
Ar
OH
O
O
BnO
BnO
BnO
+
+
NH
OH
OBn
BnO
Ar:
OBn
5a~g
6a~g
c
f
a
b
iii
H3C
O
O
d
Ar
O
BnO
BnO
BnO
BnO
O
O
O
e
BnO
BnO
N
N
Cl
O
OBn
O
OBn
Ar
O
Cl
h
8a~g
g
7a~g
i
iv
N
N
Me2N
Ar
O
N
HO
HO
HO
HO
O O
O
N
+
O
HO
HO
OH
O
OH
Ar
10a~g
9a~g
Scheme 1. Conditions and reagents: (i) diglyme, MW, 200 ꢀC, 2 min, 30–81%; (ii) Zn, 85% AcOH, rt; (iii) triphosgene, Et3N, 0 ꢀC to rt, DCM; (iv) MeOH, Pd(OH)2/C, H2.
Table 1
Ar
BnO
BnO
Ar
BnO
BnO
BnO
The yields (%) of the synthetic reaction of spiro-oxazinanone glycosides from 1 and 2
O
O
i
N
N
Run
1
Ar
3 and 4a
81.1 (1.5:1)b
7
8
9
10
O
ii
O
OBn
BnO
a
35.8
23.9
a1: 33.3
a2: 66.7
Quantitative
99.3
100
98.1
a1: 40.0
a2: 60.0
O
OBn
7a, 7c
3a, 3c
2
3
4
5
6
7
8
9
b
c
d
e
f
g
h
i
72.7 (1.2:1)
74.9 (1:1)b
75.2 (1.2:1)
68.8 (1.5:1)
43.8 (2.3:1)
41.3 (2.1:1)
35.6 (1.3:1)
37.3 (1.6:1)
34.5
26.6
28.2
28.9
35.2
34.3
28.2
26.4
23.1
19.2
15.6
15.4
97.2
95.5
96.6
99.2
98.3
O
N
BnO
BnO
BnO
BnO
O
O
O
O
N
i
ii
98.8
97.5
BnO
BnO
Ar
OBn
Ar
OBn
4a, 4c
8a, 8c
a
Calculated according to the proportions of the corresponding proton signals in
Scheme 2. Conditions and reagents: (i) Zn, 85% AcOH, rt; (ii) triphosgene, Et3N, 0 ꢀC to
rt, DCM.
the 1H NMR.
b
Isolated yields.
stereospecifically and no anomerization occurred for the in-
termediates of hemiketals (5 and 6). Consequently, the ratio of the
spiro-oxazinanone glycosides 7 and 8 was equal to that of the
derivatives 7 and 8, followed by deprotection with catalytic hy-
drogenation to afford the spiro-oxazinanone glycosides 9 and 10,
respectively.
a
- and b-anomers in the mixture of cycloadducts 3 and 4 according
It should be mentioned that the anomeric mixture of the cy-
cloaddition products 3 and 4 was very difficult to separate because
of their similar polarity, and only 3a, 4a and 3c, 4c were isolated by
repeated silica gel column chromatographic separation. With the
separated 3a, 3c, 4a, and 4c the reductive opening of the iso-
xazolidine ring and recyclization of the generated intermediates
(5a, 5c, 6a, and 6c) with triphosgene were then examined, re-
spectively (Scheme 2).
to the 1H NMR spectra.
Following the above procedures, the spiro-oxazinanone glyco-
sides 7a–g and 8a–g were synthesized from the mixtures of 3a–g
and 4a–g. However, in the case of 3h where exists a 4-pyridinyl
group, the reductive opening of its isoxazolidine ring did not give
the corresponding 1,3-amino alcoholic ketose (5h), but the C–N
bond cleaved derivatives 11h and 12h were insteadly obtained as
shown in Scheme 3. Under this reductive condition the iso-
xazolidine ring of 3h could be firstly opened at the C–N bond to
generate the hydroxylamine derivative 11h, followed by the
cleavage of the O–N bond to afford the ketose-like product 12h.
Similarly, the spiro-isoxazolidine glycoside 3i with a 2-pyridinyl
group gave product 12i under the same condition (Scheme 3). Be-
sides, various other reductive conditions and methods,10 such as
with LiAlH4 and DIBAL-H, were examined in order to cleave O–N
bond of 1,3-amino alcoholic intermediate (5h), but all the ap-
proaches were unsuccessful. The existence of pyridinyl group might
It has been found that in the two-step process of opening iso-
xazolidine ring and forming the oxazinanone ring,
and 3c were converted to their corresponding spiro-oxazinanones
7a and 7c possessing the -anomeric configuration exclusively, and
-anomers 4a and 4c afforded the sole -anomeric spiro-oxazina-
a-anomers 3a
a
b
b
nones 8a and 8c, respectively. It would be concluded that in the
process the configurations of the anomeric carbons were in re-
tention. Namely, the conversion from spiro-isoxazolidine glyco-
sides (3 and 4) to spiro-oxazinanone glycosides (7 and 8) proceeded

X. Li et al. / Tetrahedron 64 (2008) 9911–9920
9913
N
N
N
8
BnO
BnO
BnO
8
BnO
4
BnO
2
1
3
2
O
1
O
O
3
7
7
i
i
BnO
BnO
3
6
6
BnO
BnO
1
H
N
N
5 4
4
5
O
O
OBn
OH
OBn
OBn
3h
i
11h
12h
N
BnO
BnO
BnO
O
NH
OH
OBn
5h
8
6
BnO
BnO
BnO
BnO
N
4
2
N
3
O
1
O
3
7
5
i
BnO
1
N
4
O
OH
OBn
OBn
BnO
3i
12i
Scheme 3. Conditions and reagents: (i) Zn, 85% AcOH, rt.
make the C-3 of the isoxazolidine ring (an
a-C to pyridine ring)
The X-ray crystallographic structure12 of 9a1, as shown in
more electron-deficient. Consequently, the C–N bond could be
more sensitive to reductive condition than the O–N bond, and the
C–N bond could be firstly cleaved under the reductive conditions.
Subsequently, the debenzylation of the spiro-oxazinanone de-
rivatives 7a–g and 8a–g were carried out by catalytic hydrogena-
tion with Pd(OH)2/C in MeOH solution at room temperature to give
the corresponding debenzylated spiro-oxazinanone glycosides 9a–
g and 10a–g in almost quantitative yields (step (iv) in Scheme 1 and
Table 1). However, in the cases of 7a and 8a, the furan group was
also hydrogenated to form a saturated tetrahydrofuran ring to af-
ford the four diastereomers of 9a1, 9a2 and 10a1, 10a2, respectively,
as shown in Scheme 4.
The structures of all the intermediates 3a–h, 4a–h, 7a–g, 8a–g,
11, and 12, and final products 9a–g and 10a–g were determined by
the analyses of their spectral data of 1H NMR, 13C NMR, 2D-COSY
and NOESY, MS, and the X-ray crystallographic structure of 9a1
(Fig. 1). It has been reported11 that in the 13C NMR spectra of
C-glycosides and spiro-glycosides the signals of the anomeric car-
Figure 1, revealed that the anomeric (spiro) C6 possessed
0
a
-configuration, and both C4 in oxazinanone ring and C2 in tetra-
hydrofuran cycle were in S-form. Accordingly, its diastereomer
0
9a2 had S-form in the C4 and R-form at the C2 . In addition, the
D
-glucopyranosyl ring had a chair form with all OH and CH2OH
groups in equatorial position; the spiro-oxazinanone ring was in
twist-boat form perpendicular to the glucopyranose ring,
a
plane and the tetrahydrofuran cycle kept an envelope confor-
mation vertically above the oxazinanone ring. This conformation
was consistent with its NOESY correlation analysis as shown in
Figure 2(a).
The NOESY correlations of the diastereomers of 9a1 and 10a1
are shown in Figure 2(a) and (b), respectively. The NOESY correla-
tions between H5a-H4 and H5a-H7 in 9a1 (Fig. 2(a)) supported the
S-form of C4, which was identical to that in the X-ray crystallo-
graphic structure (Fig. 1). Similarly, the S-configuration at C4 in the
b-anomer (10a1) could also be assigned based on the NOESY cor-
relations between H5a-H4 and H5a-H7 (Fig. 2(b)). For the other 9 and
10, the similar NOESY correlations between H5a-H4 and H5a-H7
were observed, indicating that 9 and 10 had S-configuration at C4.
Accordingly, the key intermediates 7 and 8, and the cycloadducts
of 3 and 4 would be tentatively deduced to possess the S-configu-
ration at the corresponding carbons considering that the configu-
rations were in retention during the conversion from the
spiro-isoxazolidine (3 and 4) to the spiro-oxazinanone (7 and 8) as
shown in Scheme 1.
bon in the a-isomers appeared in upfield compared with the b-
isomers, which has been successfully used in the assignment of the
anomeric configurations of spiro-isoxazolindine C-glycosides.7,8
Accordingly, the anomeric configurations of 3a–h, 4a–h, 7a–g, 8a–g,
9a–g, and 10a–g were determined by the comparison of the
anomeric signals of the 13C NMR spectra between each two
anomeric isomers (see Table 2) with the combination of the X-ray
crystallographic structure of 9a1.
According to the X-ray crystallographic structure of 9a1 (Fig. 1),
5'
1'
2'
0
its diastereomer (9a2) had an R-form at the corresponding C2
4'
3'
H
O
(Fig. 3). In addition, the 1H NMR signals of H2 in the four isomers
O
O
0
H
O
H
11
BnO
BnO
HO
HO
HO
H
H
9a1, 9a2 and 10a1, 10a2 (Table 3) provided further useful in-
O
O
O
6
i
10
5 4
1 2
N
O
0
N
O
9
3
formation for determining the configuration of the C2 . It can be
HO
HO
N
+
8
7
seen from Table 3 that the 1H NMR signals of H2 in 9a1 and 9a2
O
O
OH
0
HO
OBn
7a
OH
9a2
O
BnO
were very similar to those in 10a1 and 10a2, which implied that 9a1
9a1
and 10a1, 9a2 and 10a2 would have the same configurations at the
0
C2 , respectively. With combination of the above analyses on the
O
N
HO
HO
HO
11
O
N
O
HO
BnO
BnO
0
O
O
S-configuration of C4, the configuration of C2 would be tentatively
O
O
1
5
O
O
2
10
i
+
9
3
6
7
HO
HO
assigned to be S-form in 10a1 and R-form in 10a2 as shown in
Figure 3.
The glycosidase inhibition and antitumor activities of com-
pounds 9 and 10 were preliminarily evaluated. The glycosidase
inhibitory activities were measured on hydrolytic reactions of
H
N
H
2'
8
4
3'
H
OH
4'
O
OH
BnO
H
OBn
O
O
H
5'
1'
8a
10a1
10a2
Scheme 4. Conditions and reagents: (i) MeOH, Pd(OH)2/C, H2, 2 h, w100%.

9914
X. Li et al. / Tetrahedron 64 (2008) 9911–9920
Figure 1. ORTEP X-ray crystallographic structure of 9a1 represented with ball and stick model (hydrogen atoms were omitted for clarity).
3. Experimental
Table 2
Chemical shifts (ppm) of the anomeric carbon of 7a–g, 8a–g, 9a–g, and 10a–g
3.1. General methods
3
4
7
8
9
10
a
105.74
107.42
100.44
101.81
a1: 102.08
a2: 101.54
101.52
101.48
101.52
101.52
101.75
101.20
a1: 102.52
a2: 102.24
102.36
102.32
102.28
102.16
102.07
101.25
Melting points were measured on a SGWÒ X-4 micro melting
point apparatus and were uncorrected. Optical rotations were de-
termined on a SGWÒ-1 automatic polarimeter. 1H NMR and 13C
NMR spectra were measured on a RT-NMR Bruker AVANCE 400
(400 MHz) NMR spectrometer using tetramethylsilane (Me4Si) as
an internal standard. Mass spectra (MS) and high resolution mass
spectra (HRMS) were carried out on a FTICR-MS (Ionspec 7.0T) mass
spectrometer with electrospray ionization (ESI). X-ray crystallo-
graphic measurements were made on a Bruker SMART CCD dif-
fractometer. The optical densities for examining the activities of
glycosidase inhibition and antitumor were measured on a TU-1901
UV–vis spectrophotometer and a BioRad Model 3550 microplate
spectrophotometer, respectively. The microwave assisted reaction
was carried out on a CEM DISCOVER S-Class Chemical Synthesis
System in a 10 mL (or 35 mL) seamless pressure vial. Thin-layer
chromatography (TLC) was performed on precoated plates (Qing-
dao GF254) with detection by UV light or with phosphomolybdic
acid in EtOH/H2O followed by heating. Column chromatography
was performed using SiO2 (Qingdao 300–400 mesh).
b
c
d
e
f
g
h
i
104.99
104.94
104.92
104.89
101.44
101.18
105.25
105.65
106.29
106.18
106.61
106.86
106.18
105.98
107.60
107.69
100.59
100.56
100.59
100.83
100.58
100.60
101.85
100.78
101.78
102.03
101.80
101.83
a
-amylase, a-glucosidase, and b-glucosidase using acarbose as
a control, respectively. As shown in Table 4, although the com-
pounds showed low inhibitory activities on the glycosidases com-
pared to the positive control, they exhibited a certain selective
inhibition to b-glucosidase. The cytotoxicity of the compounds 9
and 10 against Hela cell lines (human cervical cancer cells) was also
examined by the modified Mosmann protocol.13 It has been found
that only a few of them exhibited a slight cytotoxicity to Hela cell
lines, for instance, the inhibition of 9a1, 9a2, 9g, and 10f at the
concentration of 10 mmol/L were 12.75, 12.35, 11.09, and 15.38%,
respectively; the other compounds showed no activity against this
cell.
3.2. General procedure for the microwave assisted 1,3-dipolar
cycloaddition of exo-glucal (1) with nitrones (2a–i)
In summary, the stereoselective synthesis of a series of novel
aryl spiro-oxazinanone glucosides has been achieved by the steps
of the stereoselectively microwave assisted 1,3-dipolar cycload-
dition of exo-glucal with aryl nitrones, the reductive cleavage of
the O–N bound with Zn/AcOH/H2O, and recyclization with tri-
phosgene to afford the key intermediates of benzyl protected aryl
spiro-oxazinanone glucosides 7 and 8, which provided a conve-
nient new method for constructing spironucleoside. The catalytic
hydrogenation of 7 and 8 with Pd(OH)2/C furnished the corre-
sponding title compounds 9a–g and 10a–g quantitatively. Some
A solution of exo-glucal (2.0 mmol), nitrone (2.6 mmol) in
diglyme (2 mL) was irradiated with microwave at 200 ꢀC for 2 min.
The reaction mixture was cooled to room temperature and then
submitted to silica gel column chromatography (petroleum ether/
ethyl acetate v/v¼6:1) to purify products 3a–i and 4a–i. The results
are listed in Table 1.
3.3. General procedure for the synthesis of spiro-
oxazinanones 7 and 8
of 9 and 10 showed a certain selective inhibition against b-glu-
cosidase and a slightly antitumor activity. The further synthesis
and biological study of new spironucleosides are under way in
this lab.
A mixture of compounds 3 and 4 (0.90 mmol) and zinc (1.784 g)
in acetic acid 85% (16 mL) was vigorously stirred for 10 h. Upon
completion, the reaction mixture was neutralized with saturated aq
Na2CO3 carefully with cooling, and then extracted with EtOAc three
times. The organic phase was dried over Na2SO4 and concentrated
in vacuo to give intermediates 5 and 6 as a syrup. The mixture 5 and
6 was dissolved in 15 mL of dry dichloromethane (DCM), and Et3N
(0.72 mL) was added. The solution was stirred at 0 ꢀC for 20 min
and a solution of triphosgene (0.597 g, 0.9 mmol) in DCM (15 mL)
was added dropwise during 1 h at the same temperature. The re-
action mixture was stirred at room temperature for 12 h, then the
reaction was quenched by adding small amount of H2O. The mix-
ture was extracted with DCM two times. The organic layer was
washed with brine, dried over Na2SO4, and concentrated under
O
N
HO
O
H
O
4
Ha
6
OH
5
6
H
7
5
4
HO
7
HO
Hb
H
a Hb
H
O
(a)
(b)
Figure 2. NOESY of (a) 9a1 and (b) 10a1.

X. Li et al. / Tetrahedron 64 (2008) 9911–9920
9915
O
O
2'
S
R
O
N
O
HO
HO
HO
HO
HO
H
H
O
HO
HO
HO
HO
O
O
S
O
O
H
S
H
O
O
4
N
H
H
N
O
S
N
O
S
O
OH
R
S
OH
HO
OH
H
H
HO
HO
O
O
OH
9a2
9a1
10a1
10a2
0
Figure 3. The configurations of C4 and C2 in 9a and 10a.
Table 3
The 1H NMR signals of H2 and the configurations of C4–C2 in 9a and 10a
4.77 (d, 1H, J¼10.8 Hz, CH2Ph), 4.84 (d, 1H, J¼10.8 Hz, CH2Ph), 4.88
(d,1H, J¼10.8 Hz, CH2Ph), 5.01 (d,1H, J¼11.2 Hz, CH2Ph), 6.25 (d,1H,
J¼3.2 Hz, furyl), 6.34 (q, 1H, J¼4.0 Hz, furyl), 7.20–7.36 (m, 21H,
0
0
9a1
9a2
10a1
10a2
ArH); 13C NMR (CDCl3)
d: 28.78 (NCH3), 33.78 (5-C), 50.12 (4-C),
0
dH2
4.15 (q)
4.44 (t)
4.19 (q)
4.37 (t)
69.59 (11-C), 73.74 (10-C), 74.96 (CH2Ph), 75.49 (CH2Ph), 75.59
(CH2Ph), 76.19 (CH2Ph), 78.21 (9-C), 83.36 (7-C), 83.67 (8-C), 101.81
(6-C), 109.71 (furyl), 110.77 (furyl), 127.85 (Ar–C), 128.03 (Ar–C),
128.15 (Ar–C), 128.19 (Ar–C), 128.24 (Ar–C), 128.31 (Ar–C), 128.51
(Ar–C), 128.82 (Ar–C), 128.86 (Ar–C), 129.22 (Ar–C), 138.18 (Ar–C),
138.35 (Ar–C), 138.59 (Ar–C), 143.47 (furyl), 151.24 (furyl), 152.26
(C]O); ESI-MS: 712 [MþNa]þ.
0
Configurations of C4–C2
S–S
S–R
S–S
S–R
Table 4
Inhibitory activities of some of compounds 9 and 10 against glycosidases
Compounds
Inhibition % (in vitro)a
a
-Amylase
a
-Glucosidase
b
-Glucosidase
9a1
9a2
9b
5.41
3.32
3.51
1.98
db
7.56
db
3.3.3. Compound 7b
db
2.78
10.35
19.44
db
30
White solid, mp:151–153 ꢀC; [
a
]
þ64.04 (c 0.3, CHCl3); 1H
D
10b
3.57
5.75
db
1.30
2.87
db
9cc
NMR (CDCl3)
d
: 1.83 (dd, 1H, J¼14.0, 5.2 Hz, 5-H), 2.28 (t, 1H,
10c
9d
10d
9e
J¼13.0 Hz, 5-H), 2.69 (s, 3H, NCH3), 3.39 (d, 1H, J¼9.6 Hz, 11-H), 3.71
(d, 1H, J¼10.4 Hz, 11-H), 3.77–3.82 (m, 2H, 9-H, 7-H), 4.09 (d, 1H,
J¼9.6 Hz, 10-H), 4.33 (t, 1H, J¼9.4 Hz, 8-H), 4.51 (t, 1H, J¼2.4 Hz,
4-H), 4.54 (d, 1H, J¼2.4 Hz, CH2Ph), 4.59–4.64 (m, 3H, CH2Ph), 4.89–
4.91 (m, 3H, CH2Ph), 4.97 (d, 1H, J¼12.0 Hz, CH2Ph), 7.21–7.36 (m,
2.24
4.82
2.58
2.65
db
13.73
16.41
9.21
3.93
db
2.39
0.91
3.93
db
10e
Control (acarbose)
88.8
a
25H, ArH); 13C NMR (CDCl3)
d: 34.73 (NCH3), 37.64 (5-C), 58.07 (4-
The concentration of the inhibitor is 1.0 mg/mL.
No inhibition.
The concentration is 2.5 mg/mL.
b
c
C), 68.80 (11-C), 73.02 (CH2Ph), 73.63 (10-C), 75.04 (CH2Ph), 75.84
(CH2Ph), 76.22 (CH2Ph), 78.23 (9-C), 81.94 (7-C), 82.71 (8-C), 100.59
(6-C), 127.30 (Ar–C), 127.70 (Ar–C), 127.91 (Ar–C), 128.06 (Ar–C),
128.16 (Ar–C), 128.36 (Ar–C), 128.72 (Ar–C), 128.79 (Ar–C), 129.43
(Ar–C), 138.15 (Ar–C), 138.40 (Ar–C), 138.82 (Ar–C), 140.17 (Ar–C),
152.99 (C]O); ESI-MS: 722 [MþNa]þ.
reduced pressure. The residue was applied to silica gel column
chromatography (petroleum ether/ethyl acetate v/v¼5:1) to afford
the products 7 and 8. From the mixture, spiro-oxazinanone glu-
cosides 7 and 8 were isolated by repeated column chromatographic
separation (cyclohexane/ethyl acetate v/v¼8:1). The results are
shown in Table 1.
3.3.4. Compound 8b
30
Pale yellow syrup; [
a
]
þ68.56 (c 0.4, CHCl3); 1H NMR (CDCl3)
d:
D
2.35–2.46 (m, 2H, 5-H), 2.73 (s, 3H, NCH3), 3.65–3.71 (m, 4H, 7-H,
11-H, 9-H), 3.80–3.85 (m, 2H, 10-H, 8-H), 4.46 (dd, 1H, J¼6.0,
10.8 Hz, 4-H), 4.58–4.89 (m, 7H, CH2Ph), 4.99 (d, 1H, J¼11.2 Hz,
3.3.1. Compound 7a
25
White solid, mp: 193–194 ꢀC; [
a
]
þ41.53 (c 0.4, CHCl3); 1H
CH2Ph), 7.07–7.39 (m, 25H, ArH); 13C NMR (CDCl3)
d: 32.99 (NCH3),
D
NMR (CDCl3)
d
: 1.74 (dd, 1H, J¼13.8, 4.6 Hz, 5-H), 2.57 (t, 1H,
34.70 (5-C), 57.53 (4-C), 69.63 (11-C), 73.76 (CH2Ph), 75.03 (10-C),
75.51 (CH2Ph), 75.56 (CH2Ph), 76.13 (CH2Ph), 78.28 (9-C), 83.36 (7-
C), 83.77 (8-C), 101.85 (6-C), 127.27 (Ar–C), 127.86 (Ar–C), 128.02
(Ar–C), 128.13 (Ar–C), 128.25 (Ar–C), 128.36 (Ar–C), 128.80 (Ar–C),
128.88 (Ar–C), 129.53 (Ar–C), 138.17 (Ar–C), 138.46 (Ar–C), 138.59
(Ar–C), 138.63 (Ar–C), 139.80 (Ar–C), 153.15 (C]O); ESI-MS: 700
[MþH]þ.
J¼13.0 Hz, 5-H), 2.73 (s, 3H, NCH3), 3.40 (d, 1H, J¼9.6 Hz, 7-H), 3.66
(d, 1H, J¼10.8 Hz, 11-H), 3.77 (d, 1H, J¼3.2 Hz, 11-H), 3.80 (t, 1H,
J¼9.6 Hz, 9-H), 4.04–4.06 (m, 1H, 10-H), 4.32 (t, 1H, J¼10.2 Hz, 8-H),
4.50 (d, 1H, J¼12.0 Hz, CH2Ph), 4.58–4.69 (m, 4H, 4-H, CH2Ph), 4.89
(d, 1H, J¼9.6 Hz, CH2Ph), 4.91 (s, 2H, CH2Ph), 5.00 (d, 1H, J¼12.0 Hz,
CH2Ph), 6.26 (d, 1H, J¼3.2 Hz, furyl), 6.36 (q, 1H, J¼3.2 Hz, furyl),
7.20–7.42 (m, 21H, ArH); 13C NMR (CDCl3)
d: 33.44 (NCH3), 33.80 (5-
C), 50.71 (4-C), 68.69 (11-C), 72.97 (CH2Ph), 73.65 (10-C), 75.04
(CH2Ph), 75.79 (CH2Ph), 76.23 (CH2Ph), 78.15 (9-C), 81.60 (7-C),
82.69 (8-C), 100.44 (6-C), 109.51 (furyl), 109.70 (furyl), 127.69 (Ar–
C), 127.92 (Ar–C), 128.09 (Ar–C), 128.17 (Ar–C), 128.23 (Ar–C),
128.39 (Ar–C), 128.54 (Ar–C), 128.73 (Ar–C), 128.79 (Ar–C), 138.16
(Ar–C), 138.36 (Ar–C), 138.80 (Ar–C), 143.30 (furyl), 151.55 (furyl),
152.15 (C]O); ESI-MS: 712 [MþNa]þ.
3.3.5. Compound 7c
30
Colorless syrup; [
a
]
þ69.18 (c 1.3, CHCl3); 1H NMR (CDCl3)
D
d: 1.87 (dd, 1H, J¼14.0, 5.2 Hz, 5-H), 2.31 (t, 1H, J¼13.0 Hz, 5-H),
2.39 (s, 3H, ArCH3), 2.69 (s, 3H, NCH3), 3.39 (d, 1H, J¼9.6 Hz,
7-H), 3.70 (d, 1H, J¼10.4 Hz, 11-H), 3.79–3.83 (m, 2H, 9-H, 11-H),
4.09 (d, 1H, J¼9.6 Hz, 10-H), 4.33 (t, 1H, J¼9.4 Hz, 8-H), 4.49–4.52
(m, 1H, 4-H), 4.54 (d, 1H, J¼12.4 Hz, CH2Ph), 4.61 (d, 1H,
J¼10.8 Hz, CH2Ph), 4.64 (d, 1H, J¼12.0 Hz, CH2Ph), 4.90 (d, 1H,
J¼10.8 Hz, CH2Ph), 4.91 (s, 2H, CH2Ph), 4.97 (d, 1H, J¼11.6 Hz,
3.3.2. Compound 8a
30
Colorless syrup; [
a
]
þ10.13 (c 2.5, CHCl3); 1H NMR (CDCl3)
d:
CH2Ph), 7.09–7.37 (m, 24H, ArH); 13C NMR (CDCl3)
d: 21.51
D
2.37 (dd,1H, J¼14.8, 4.4 Hz, 5-H), 2.67–2.70 (m,1H, 5-H), 2.74 (s, 3H,
NCH3), 3.63–3.65 (m, 4H, 7-H, 11-H, 9-H), 3.78–3.79 (m, 2H, 10-H,
8-H), 4.54–4.60 (m, 4H, 4-H, CH2Ph), 4.75 (d, 1H, J¼10.8 Hz, CH2Ph),
(ArCH3), 34.63 (NCH3), 37.56 (5-C), 57.68 (4-C), 68.71 (11-C),
72.94 (CH2Ph), 73.61 (10-C), 75.00 (CH2Ph), 75.80 (CH2Ph), 76.19
(CH2Ph), 78.15 (9-C), 81.98 (7-C), 82.66 (8-C), 100.56 (6-C),

9916
X. Li et al. / Tetrahedron 64 (2008) 9911–9920
126.90 (Ar–C), 127.26 (Ar–C), 127.66 (Ar–C), 127.88 (Ar–C), 128.04
(Ar–C), 128.11 (Ar–C), 128.15 (Ar–C), 128.22 (Ar–C), 128.35 (Ar–
C), 128.70 (Ar–C), 128.71 (Ar–C), 128.77 (Ar–C), 129.67 (Ar–C),
130.07 (Ar–C), 137.02 (Ar–C), 138.14 (Ar–C), 138.35 (Ar–C), 138.49
(Ar–C), 138.79 (Ar–C), 152.99 (C]O); ESI-MS: 713 [M]þ.
132.77 (Ar–C), 137.63 (Ar–C), 138.19 (Ar–C), 138.25 (Ar–C), 138.60
(Ar–C), 138.87 (Ar–C), 152.91 (C]O); ESI-MS: 733 [M]þ.
3.3.10. Compound 8e
30
Colorless syrup; [
a
]
þ23.61 (c 0.8, CHCl3); 1H NMR (CDCl3)
d:
D
2.21 (m, 1H, 5-H), 2.56 (m, 1H, 5-H), 2.75 (s, 3H, NCH3), 3.67–3.85
(m, 6H, 7-H, 8-H, 9-H, 10-H, 11-H), 4.58–4.65 (m, 3H, 4-H, CH2Ph),
4.76–4.99 (m, 6H, CH2Ph), 7.16–7.40 (m, 24H, ArH); 13C NMR (CDCl3)
3.3.6. Compound 8c
23
White solid, mp: 98–100 ꢀC; [
a
]
þ71.56 (c 3.1, CHCl3); 1H
D
d: 30.89 (NCH3), 35.02 (5-C), 53.85 (4-C), 69.43 (11-C), 73.72
NMR (CDCl3)
d
: 1.85 (dd, 1H, J¼14.0, 5.2 Hz, 5-H), 2.30 (t, 1H,
(CH2Ph), 75.11 (10-C), 75.60 (CH2Ph), 75.71 (CH2Ph), 76.23 (CH2Ph),
78.25 (9-C), 83.39 (7-C), 83.85 (8-C), 102.03 (6-C), 126.85 (Ar–C),
127.91 (Ar–C), 127.96 (Ar–C), 128.15 (Ar–C), 128.22 (Ar–C), 128.40
(Ar–C), 128.52 (Ar–C), 128.79 (Ar–C), 128.82 (Ar–C), 128.92 (Ar–C),
129.42 (Ar–C), 129.72 (Ar–C), 130.33 (Ar–C), 133.10 (Ar–C), 137.15
(Ar–C), 138.13 (Ar–C), 138.42 (Ar–C), 138.64 (Ar–C), 153.56 (C]O);
ESI-MS: 756 [MþNa]þ.
J¼13.0 Hz, 5-H), 2.37 (s, 3H, ArCH3), 2.68 (s, 3H, NCH3), 3.37 (d,
1H, J¼9.6 Hz, 11-H), 3.68 (d, 1H, J¼9.6 Hz, 11-H), 3.76–3.80 (m,
2H, 9-H, 7-H), 4.09 (d, 1H, J¼9.6 Hz, 10-H), 4.32 (t, 1H, J¼9.6 Hz,
8-H), 4.48–4.53 (m, 2H, 4-H, CH2Ph), 4.58–4.63 (m, 3H, CH2Ph),
4.88–4.89 (m, 3H, CH2Ph), 4.95 (d, 1H, J¼12.0 Hz, CH2Ph), 7.02–
7.35 (m, 24H, ArH); 13C NMR (CDCl3)
d: 21.69 (ArCH3), 34.82
(NCH3), 37.84 (5-C), 57.93 (4-C), 69.00 (11-C), 73.20 (CH2Ph),
73.83 (10-C), 75.22 (CH2Ph), 76.04 (CH2Ph), 76.39 (CH2Ph), 78.44
(9-C), 82.27 (7-C), 82.89 (8-C), 100.78 (6-C), 127.48 (Ar–C), 127.89
(Ar–C), 128.09 (Ar–C), 128.32 (Ar–C), 128.36 (Ar–C), 128.44 (Ar–
C), 128.54 (Ar–C), 128.92 (Ar–C), 128.98 (Ar–C), 130.28 (Ar–C),
137.27 (Ar–C), 138.39 (Ar–C), 138.61 (Ar–C), 138.71 (Ar–C), 139.03
(Ar–C), 153.21 (C]O); ESI-MS: 714 [MþH]þ.
3.3.11. Compound 7f
20
Colorless syrup; [
a
]
þ58.74 (c 3.0, CHCl3); 1H NMR (CDCl3)
d:
D
1.78 (dd, 1H, J¼14.0, J¼5.2 Hz, 5-H), 2.21 (t, 1H, J¼13.2 Hz, 5-H), 2.67
(s, 3H, NCH3), 3.36 (d, 1H, J¼9.6 Hz, 11-H), 3.66 (d, 1H, J¼11.2 Hz, 11-
H), 3.75–3.80 (m, 2H, 9-H, 7-H), 4.06 (d, 1H, J¼9.6 Hz,10-H), 4.30 (d,
1H, J¼9.6 Hz, 8-H), 4.41 (dd, 1H, J¼12.0, 5.2 Hz, 4-H), 4.48 (d, 1H,
J¼12.0 Hz, CH2Ph), 4.56–4.61 (m, 3H, CH2Ph), 4.86–4.88 (m, 3H,
CH2Ph), 4.95 (d, 1H, J¼12.0 Hz, CH2Ph), 5.95 (s, 2H, CH2), 6.59–6.64
(m, 2H, ArH), 6.75 (d, 1H, J¼8.4 Hz, ArH), 7.18–7.33 (m, 20H, ArH);
3.3.7. Compound 7d
30
Colorless syrup; [
a
]
þ64.61 (c 2.2, CHCl3); 1H NMR (CDCl3)
d:
D
13C NMR (CDCl3)
d: 34.53 (NCH3), 37.58 (5-C), 57.80 (4-C), 68.81 (11-
1.80 (dd, 1H, J¼14.0, 5.2 Hz, 5-H), 2.21 (t, 1H, J¼12.0 Hz, 5-H), 2.70
(s, 3H, NCH3), 3.41 (d, 1H, J¼9.6 Hz, 11-H), 3.71 (d, 1H, J¼10.4 Hz, 11-
H), 3.80–3.84 (m, 2H, 9-H, 7-H), 4.11 (d, 1H, J¼9.6 Hz, 10-H), 4.35 (t,
1H, J¼9.4 Hz, 8-H), 4.50–4.55 (m, 2H, 4-H, CH2Ph), 4.60–4.65 (m,
3H, CH2Ph), 4.91–4.93 (m, 3H, CH2Ph), 4.99 (d, 1H, J¼12.0 Hz,
C), 73.04 (CH2Ph), 73.66 (10-C), 75.08 (CH2Ph), 75.91 (CH2Ph), 76.25
(CH2Ph), 78.26 (9-C), 82.04 (7-C), 82.74 (8-C), 100.58 (6-C), 101.77
(CH2), 107.31 (Ar–C), 108.87 (Ar–C), 121.04 (Ar–C), 127.75 (Ar–C),
127.96 (Ar–C), 128.12 (Ar–C), 128.21 (Ar–C), 128.41 (Ar–C), 128.76
(Ar–C), 128.83 (Ar–C), 133.92 (Ar–C), 138.23 (Ar–C), 138.42 (Ar–C),
138.85 (Ar–C), 148.06 (Ar–C), 148.83 (Ar–C), 152.92 (C]O); ESI-MS:
744 [MþH]þ.
CH2Ph), 7.13–7.38 (m, 24H, ArH); 13C NMR (CDCl3)
d: 34.77 (NCH3),
37.59 (5-C), 57.56 (4-C), 68.82 (11-C), 73.11 (CH2Ph), 73.68 (10-C),
75.11 (CH2Ph), 75.90 (CH2Ph), 76.27 (CH2Ph), 78.24 (9-C), 81.85 (7-
C), 82.73 (8-C), 100.59 (6-C), 127.76 (Ar–C), 127.98 (Ar–C), 128.14
(Ar–C), 128.22 (Ar–C), 128.29 (Ar–C), 128.39 (Ar–C), 128.49 (Ar–C),
128.67 (Ar–C), 128.77 (Ar–C), 129.72 (Ar–C), 134.52 (Ar–C), 138.14
(Ar–C), 138.38 (Ar–C), 138.80 (Ar–C), 152.91 (C]O); ESI-MS: 734
[MþH]þ.
3.3.12. Compound 8f
20
Colorless syrup; [
a
]
ꢁ26.25 (c 0.8, CHCl3); 1H NMR (CDCl3)
d:
D
2.28–2.38 (m, 2H, 5-H), 2.70 (s, 3H, NCH3), 3.63–3.66 (m, 4H, 7-H,
11-H, 9-H), 3.75–3.80 (m, 2H, 10-H, 8-H), 4.34 (dd, 1H, J¼6.0,
10.8 Hz, 4-H), 4.53–4.61 (m, 3H, CH2Ph), 4.75–4.86 (m, 4H, CH2Ph),
4.96 (d, 1H, J¼11.2 Hz, CH2Ph), 5.96 (m, 2H, CH2), 6.62–6.65 (m, 2H,
ArH), 6.74 (d, 1H, J¼8.0 Hz, ArH), 7.24–7.35 (m, 20H, ArH); 13C NMR
3.3.8. Compound 8d
22
Colorless syrup; [
a]
þ63.02 (c 0.3, CHCl3); 1H NMR (CDCl3)
d:
D
(CDCl3) d: 32.88 (NCH3), 34.48 (5-C), 57.22 (4-C), 69.62 (11-C), 73.74
2.29–2.43 (m, 2H, 5-H), 2.70 (s, 3H, NCH3), 3.63–3.69 (m, 4H, 7-H,
11-H, 9-H), 3.79–3.84 (m, 2H, 10-H, 8-H), 4.44 (dd, 1H, J¼5.2,
12.0 Hz, 4-H), 4.57–4.63 (m, 3H, CH2Ph), 4.75–4.92 (m, 4H, CH2Ph),
4.97 (d, 1H, J¼11.2 Hz, CH2Ph), 7.12–7.38 (m, 24H, ArH); 13C NMR
(CH2Ph), 75.01 (10-C), 75.55 (CH2Ph), 76.15 (CH2Ph), 78.27 (9-C),
83.37 (7-C), 83.80 (8-C), 101.80 (6-C, CH2), 107.22 (Ar–C), 108.90
(Ar–C), 120.99 (Ar–C), 127.87 (Ar–C), 128.04 (Ar–C), 128.17 (Ar–C),
128.27 (Ar–C), 128.39 (Ar–C), 128.83 (Ar–C), 128.90 (Ar–C), 133.54
(Ar–C), 138.17 (Ar–C), 138.46 (Ar–C), 138.60 (Ar–C), 138.63 (Ar–C),
148.15 (Ar–C), 148.89 (Ar–C), 153.05 (C]O); ESI-MS: 766 [MþNa]þ.
(CDCl3) d: 32.98 (NCH3), 34.69 (5-C), 56.98 (4-C), 69.64 (11-C), 73.75
(CH2Ph), 75.10 (10-C), 75.52 (CH2Ph), 75.57 (CH2Ph), 76.13 (CH2Ph),
78.25 (9-C), 83.34 (7-C), 83.73 (8-C), 101.78 (6-C), 127.86 (Ar–C),
128.06 (Ar–C), 128.18 (Ar–C), 128.24 (Ar–C), 128.34 (Ar–C), 128.39
(Ar–C), 128.63 (Ar–C), 128.82 (Ar–C), 129.76 (Ar–C), 134.67 (Ar–C),
138.11 (Ar–C), 138.37 (Ar–C), 138.54 (Ar–C), 153.00 (C]O); ESI-MS:
756 [MþNa]þ.
3.3.13. Compound 7g
Pale yellow syrup;
(CDCl3)
22
[
a
]
þ71.05 (c 1.2, CHCl3); 1H NMR
D
d
:
1.87 (dd, 1H, J¼14.0, 5.2 Hz, 5-H), 2.34 (t, 1H,
J¼12.0 Hz, 5-H), 2.67 (s, 3H, NCH3), 2.95 (s, 6H, NCH3), 3.37 (d,
1H, J¼9.6 Hz, 11-H), 3.67 (d, 1H, J¼10.8 Hz, 11-H), 3.76–3.81 (m,
2H, 9-H, 7-H), 3.10 (m, 1H, 10-H), 4.31 (t, 1H, J¼9.6 Hz, 8-H),
4.46–4.62 (m, 5H, 4-H, CH2Ph), 4.86–4.89 (m, 3H, CH2Ph), 4.94
(d, 1H, J¼12.0 Hz, CH2Ph), 6.68 (d, 2H, J¼8.4 Hz, ArH), 7.05 (d,
3.3.9. Compound 7e
21
White solid, mp: 53–55 ꢀC; [
a
]
þ81.51 (c 1.0, CHCl3); 1H NMR
D
(CDCl3) d: 2.12 (m, 1H, 5-H), 2.73 (m, 1H, 5-H), 2.76 (s, 3H, NCH3),
3.49 (d, 1H, J¼9.6 Hz, 11-H), 3.73–3.90 (m, 4H, 7-H, 9-H, 10-H, 11-H),
2H, J¼8.4 Hz, ArH), 7.18–7.33 (m, 20H, ArH); 13C NMR (CDCl3)
d:
4.22 (m, 1H, 8-H), 4.60–5.10 (m, 9H, 4-H, CH2Ph), 7.12–7.45 (m, 24H,
34.46 (NCH3), 37.56 (5-C), 40.91 (NCH3), 57.35 (4-C), 68.82 (11-
C), 72.97 (CH2Ph), 73.65 (10-C), 75.04 (CH2Ph), 75.88 (CH2Ph),
76.21 (CH2Ph), 78.29 (9-C), 82.23 (7-C), 82.74 (8-C), 100.60 (6-C),
113.03 (Ar–C), 127.15 (Ar–C), 127.72 (Ar–C), 127.92 (Ar–C), 128.07
(Ar–C), 128.14 (Ar–C), 128.20 (Ar–C), 128.38 (Ar–C), 128.78 (Ar–
C), 128.82 (Ar–C), 138.30 (Ar–C), 138.47 (Ar–C), 138.91 (Ar–C),
150.89 (Ar–C), 153.11 (C]O); ESI-MS: 765 [MþNa]þ.
ArH); 13C NMR (CDCl3)
d: 32.37 (NCH3), 36.51 (5-C), 56.53 (4-C),
68.68 (11-C), 73.41 (CH2Ph), 73.66 (10-C), 75.16 (CH2Ph), 75.86
(CH2Ph), 75.93 (CH2Ph), 78.31 (9-C), 81.93 (7-C), 82.79 (8-C), 100.83
(6-C), 126.90 (Ar–C), 127.86 (Ar–C), 128.03 (Ar–C), 128.12 (Ar–C),
128.24 (Ar–C), 128.33 (Ar–C), 128.41 (Ar–C), 128.51 (Ar–C), 128.78
(Ar–C), 128.86 (Ar–C), 129.09 (Ar–C), 129.30 (Ar–C), 130.25 (Ar–C),

X. Li et al. / Tetrahedron 64 (2008) 9911–9920
9917
3.3.14. Compound 8g
3.4.4. Compound 10a2
White solid, mp: 129–131 ꢀC; [
(D2O) d
: 1.56–1.61 (m, 1H, 30-H), 1.83–1.88 (m, 2H, 40-H), 1.95–2.01
22
21
Pale yellow syrup; [
a]
ꢁ40.59 (c 1.0, CHCl3); 1H NMR
a
]
þ58.68 (c 0.3, H2O); 1H NMR
D
D
(CDCl3)
d: 2.33–2.38 (m, 2H, 5-H), 2.68 (s, 3H, NCH3), 2.95 (s, 6H,
NCH3), 3.61–3.68 (m, 4H, 7-H, 11-H, 9-H), 3.75–3.81 (m, 2H, 10-
H, 8-H), 4.33 (dd, 1H, J¼6.0, 10.8 Hz, 4-H), 4.53–4.85 (m, 7H,
CH2Ph), 4.99 (d, 1H, J¼11.2 Hz, CH2Ph), 6.67 (d, 2H, J¼8.4 Hz,
ArH), 7.03 (d, 2H, J¼8.4 Hz, ArH), 7.19–7.36 (m, 20H, ArH); 13C
(m, 2H, 5-H, 30-H), 2.26 (dd, 1H, J¼15.2, 5.2 Hz, 5-H), 2.89 (s, 3H,
NCH3), 3.37–3.48 (m, 4H, 7-H, 10-H, 11-H), 3.55–3.61 (m, 2H, 50-H,
4-H), 3.70–3.81 (m, 3H, 8-H, 9-H, 50-H), 4.37 (t, 1H, J¼7.2 Hz, 20-H);
13C NMR (D2O) : 20.55 (40-C), 25.88 (30-C), 26.95 (NCH3), 33.43 (5-
d
NMR (CDCl3)
d: 32.74 (NCH3), 34.40 (5-C), 40.88 (NCH3), 56.76
C), 54.51 (4-C), 61.03 (50-C), 69.66 (11-C), 69.76 (10-C), 73.69 (9-C),
74.72 (7-C), 75.64 (8-C), 77.00 (20-C), 102.24 (6-C), 155.30 (C]O);
HRMS (ESI): calcd for C14H23NO8 (Mþ): 333.1424, found: 333.1428.
(4-C), 69.63 (11-C), 73.74 (CH2Ph), 74.91 (10-C), 75.42 (CH2Ph),
75.55 (CH2Ph), 76.13 (CH2Ph), 78.31 (9-C), 83.35 (7-C), 83.82 (8-
C), 101.83 (6-C), 113.04 (Ar–C), 126.77 (Ar–C), 127.86 (Ar–C),
127.98 (Ar–C), 128.04 (Ar–C), 128.12 (Ar–C), 128.23 (Ar–C), 128.30
(Ar–C), 128.38 (Ar–C), 128.79 (Ar–C), 128.88 (Ar–C), 138.23 (Ar–
C), 138.60 (Ar–C), 138.69 (Ar–C), 150.96 (Ar–C), 153.20 (C]O);
ESI-MS: 765 [MþNa]þ.
3.4.5. Compound 9b
20
White solid, mp: 226–228 ꢀC; [
a
]
þ39.41 (c 0.5, H2O); 1H NMR
D
(D2O) d: 2.20–2.24 (m, 1H, 5-H), 2.38–2.45 (m, 1H, 5-H), 2.64 (s, 3H,
NCH3), 3.37–3.52 (m, 2H, 11-H), 3.71–3.82 (m, 4H, 10-H, 9-H, 8-H 7-
H), 4.74–4.78 (m,1H, 4-H), 7.37–7.43 (m, 5H, ArH); 13C NMR (D2O)
d
:
34.51 (NCH3), 36.76 (5-C), 57.74 (4-C), 60.76 (11-C), 69.63 (10-C),
73.24 (9-C), 73.73 (7-C), 74.79 (8-C), 101.52 (6-C), 127.75 (Ar–C),
129.11 (Ar–C), 129.71 (Ar–C), 139.57 (Ar–C), 155.58 (C]O); HRMS
3.4. General procedure for the catalytic hydrogenation
of 7 and 8
(ESI): calcd for
362.1222.
C
16H21NO7Na ([MþNa]þ): 362.1216, found:
To a solution of 7 or 8 (150 mg) in MeOH (10 mL) was added
Pd(OH)2/C catalyst (100 mg) and the mixture was vigorously stirred
under H2 atmosphere for 2 h. The catalyst was removed by filtra-
tion, the filtrate was concentrated in vacuo, and the residue was
purified by silica gel column chromatography using ethyl acetate/
MeOH (5:1) as the eluent to give the corresponding spiro-oxazi-
nanone glucosides 9 or 10 (almost quantitatively), respectively. In
the cases of 7a and 8a, the furyl-hydrogenated products 9a1, 9a2
and 10a1, 10a2 were quantitatively obtained. The results are listed
in Table 1.
3.4.6. Compound 10b
White solid, mp: 184–186 ꢀC; [
(D2O)
21
a]
ꢁ43.62 (c 0.5, H2O); 1H NMR
D
d
: 2.24 (d, 1H, J¼14.4 Hz, 5-H), 2.50 (dd, 1H, J¼10.4, 5.2 Hz, 5-
H), 2.56 (s, 3H, NCH3), 3.41–3.55 (m, 4H, 11-H, 10-H, 7-H), 3.67 (dd,
1H, J¼10.4, 5.2 Hz, 9-H), 3.85 (d, 1H, J¼12.4 Hz, 8-H), 4.48 (dd, 1H,
J¼8.4, 4.8 Hz 4-H), 7.28–7.39 (m, 5H, ArH); 13C NMR (D2O)
d: 30.55
(NCH3), 34.45 (5-C), 57.00 (4-C), 61.24 (11-C), 69.88 (10-C), 73.75 (9-
C), 74.73 (7-C), 75.81 (8-C), 102.36 (6-C), 127.84 (Ar–C), 129.20 (Ar–
C), 129.69 (Ar–C), 139.12 (Ar–C), 155.36 (C]O); HRMS (ESI): calcd
for C16H22NO7 ([MþH]þ): 340.1396, found: 340.1408.
3.4.1. Compound 9a1
22
White solid, mp: 222–224 ꢀC; [
a
]
þ62.34 (c 0.3, H2O); 1H NMR
D
(D2O)
d
: 1.52–1.56 (m, 1H, 30-H), 1.81–1.85 (m, 2H, 40-H), 1.89–2.01
3.4.7. Compound 9c
21
(m, 2H, 5-H, 30-H), 2.07–2.13 (m, 1H, 5-H), 2.92 (s, 3H, NCH3), 3.31
(d, 1H, J¼9.6 Hz, 11-H), 3.38 (t, 1H, J¼9.6 Hz, 11-H), 3.52–3.55 (m,
1H, 10-H), 3.60–3.80 (m, 6H, 7-H, 8-H, 9-H, 50-H, 4-H), 4.15 (q, 1H,
White solid, mp: 227–229 ꢀC; [
a]
þ108.68 (c 0.5, H2O); 1H
D
NMR (D2O) d: 2.15–2.19 (m, 1H, 5-H), 2.30 (s, 3H, CH3), 2.36–2.43
(m, 1H, 5-H), 2.63 (s, 3H, NCH3), 3.38–3.43 (m, 1H, 11-H), 3.45–3.53
(m, 1H, 11-H), 3.71–3.84 (m, 4H, 10-H, 9-H, 8-H, 7-H), 4.74–4.76 (m,
J¼14.4, 7.2 Hz, 20-H); 13C NMR (D2O)
d
: 25.57 (40-C), 26.78 (30-C),
29.20 (NCH3), 35.01 (5-C), 55.37 (4-C), 60.57 (50-C), 69.29 (11-C),
69.47 (10-C), 73.18 (9-C), 73.82 (7-C), 74.68 (8-C), 80.18 (20-C),
102.08 (6-C), 155.32 (C]O); HRMS (ESI): calcd for C14H23NO8 (Mþ):
333.1424, found: 333.1422.
1H, 4-H), 7.24–7.25 (m, 4H, ArH); 13C NMR (D2O)
d: 20.64 (CH3),
34.40 (NCH3), 36.75 (5-C), 57.41 (4-C), 60.77 (11-C), 69.65 (10-C),
73.24 (9-C), 73.74 (7-C), 74.77 (8-C), 101.48 (6-C), 127.75 (Ar–C),
130.21 (Ar–C), 136.49 (Ar–C), 139.42 (Ar–C), 155.54 (C]O); HRMS
(ESI): calcd for C17H23NO7Na ([MþNa]þ): 376.1372, found: 376.1377.
3.4.2. Compound 9a2
22
White solid, mp: 174–176 ꢀC; [
a]
þ28.67 (c 0.3, H2O); 1H NMR
3.4.8. Compound 10c
D
22
(D2O)
d
: 1.62–1.69 (m, 1H, 30-H), 1.91–1.98 (m, 2H, 30-H, 40-H), 2.03–
White solid, mp: 142–144 ꢀC; [
a
]
ꢁ16.81 (c 0.5, H2O); 1H NMR
D
2.10 (m, 2H, 5-H, 40-H), 2.30 (t, 1H, J¼12.0 Hz, 5-H), 3.02 (s, 3H,
NCH3), 3.43 (d, 1H, J¼9.6 Hz, 11-H), 3.49 (t, 1H, J¼9.6 Hz, 11-H),
3.62–3.65 (m,1H,10-H), 3.71–3.92 (m, 6H, 7-H, 8-H, 9-H, 50-H, 4-H),
(D2O) d: 2.29 (m, 4H, 5-H, CH3), 2.50 (m, 1H, 5-H), 2.60 (s, 3H,
NCH3), 3.49–3.60 (m, 4H, 11-H, 10-H, 9-H), 3.73–3.74 (m, 1H, 7-H),
3.89–3.91 (m, 1H, 8-H), 4.50–4.51 (m, 1H, 4-H), 7.24 (m, 4H, ArH);
4.44 (t, 1H, J¼7.2 Hz, 20-H); 13C NMR (D2O)
d
: 25.87 (40-C), 26.94 (30-
13C NMR (D2O)
d: 20.63 (CH3), 30.50 (NCH3), 34.33 (5-C), 56.65 (4-
C), 27.10 (NCH3), 33.72 (5-C), 55.40 (4-C), 60.66 (50-C), 69.60 (11-C),
69.68 (10-C), 73.23 (9-C), 73.86 (7-C), 74.68 (8-C), 77.26 (20-C),
101.54 (6-C), 155.50 (C]O); HRMS (ESI): calcd for C14H23NO8Na:
356.1321, found: 356.1327.
C), 61.21 (11-C), 69.86 (10-C), 73.73 (9-C), 74.70 (7-C), 75.77 (8-C),
102.32 (6-C), 127.82 (Ar–C), 130.18 (Ar–C), 136.03 (Ar–C), 139.53
(Ar–C), 155.32 (C]O); HRMS (ESI): calcd for
C17H23NO7Na
([MþNa]þ): 376.1372, found: 376.1381.
3.4.3. Compound 10a1
3.4.9. Compound 9d
20
22
White solid, mp: 186–188 ꢀC; [
a
]
ꢁ15.82 (c 0.4, CH3OH); 1H
Pale yellow solid, mp: 239–241 ꢀC; [
a
]
þ18.41 (c 0.5, H2O);
D
D
NMR (D2O)
d
: 1.55–1.61 (m, 1H, 30-H), 1.79–1.89 (m, 2H, 30-H, 40-H),
1H NMR (D2O)
d: 2.20–2.24 (m, 1H, 5-H), 2.39–2.46 (m, 1H, 5-H),
1.91–1.98 (m, 2H, 5-H, 40-H), 2.22 (dd, 1H, J¼15.2, 5.2 Hz, 5-H), 2.89
(s, 3H, NCH3), 3.34–3.46 (m, 4H, 7-H, 10-H, 11-H), 3.57–3.64 (m, 2H,
50-H, 4-H), 3.70–3.78 (m, 3H, 8-H, 9-H, 50-H), 4.19 (q, 1H, J¼14.4,
2.65 (s, 3H, NCH3), 3.41 (d, 1H, J¼9.6 Hz, 11-H), 3.50 (t, 1H,
J¼8.0 Hz, 11-H), 3.71–3.88 (m, 4H, 10-H, 9-H, 8-H 7-H), 4.72–
4.81 (m, 1H, 4-H), 7.37–7.45 (m, 4H, ArH); 13C NMR (D2O)
d:
7.2 Hz, 20-H); 13C NMR (D2O)
d
: 22.16 (40-C), 25.68 (30-C), 26.50
34.51 (NCH3), 36.76 (5-C), 57.73 (4-C), 60.76 (11-C), 69.63 (10-C),
73.23 (9-C), 73.71 (7-C), 74.78 (8-C), 101.52 (6-C), 127.75 (Ar–C),
129.10 (Ar–C), 129.70 (Ar–C), 139.55 (Ar–C), 155.57 (C]O);
(NCH3), 34.41 (5-C), 54.17 (4-C), 61.04 (50-C), 69.40 (11-C), 69.77
(10-C), 73.69 (9-C), 74.67 (7-C), 75.64 (8-C), 79.54 (20-C), 102.52 (6-
C), 155.15 (C]O); HRMS (ESI): calcd for C14H24NO8 ([MþH]þ):
334.1502, found: 334.1509.
HRMS (ESI): calcd for
found: 396.0829.
C
16H20ClNO7Na ([MþNa]þ): 396.0826,

9918
X. Li et al. / Tetrahedron 64 (2008) 9911–9920
3.4.10. Compound 10d
(4-C), 61.44 (11-C), 70.36 (10-C), 73.67 (9-C), 74.45 (7-C), 75.32 (8-
25
White solid, mp: 199–201 ꢀC; [
a
]
D
þ60.58 (c 0.8, H2O); 1H
C), 101.20 (6-C), 113.37 (Ar–C), 127.39 (Ar–C), 128.12 (Ar–C), 151.18
(Ar–C), 154.47 (C]O); HRMS (ESI): calcd for C18H27N2O7 ([MþH]þ):
383.1818, found: 383.1824.
NMR (D2O) d: 2.24–2.30 (m, 1H, 5-H), 2.49–2.52 (m, 1H, 5-H),
2.66 (s, 3H, NCH3), 3.24–3.29 (m, 1H, 11-H), 3.45–3.57 (m, 3H,
10-H, 11-H, 7-H), 3.69–3.78 (m, 1H, 9-H), 3.87–3.90 (m, 1H, 8-H),
4.49–4.53 (m, 1H, 4-H), 7.19–7.39 (m, 4H, ArH); 13C NMR (D2O)
3.4.16. Compound 10g
21
d: 30.58 (NCH3), 34.33 (5-C), 57.00 (4-C), 61.27 (11-C), 69.91 (10-
White solid, mp: 177–179 ꢀC; [
a
]
þ50.68 (c 0.7, H2O); 1H
D
C), 73.80 (9-C), 74.71 (7-C), 75.84 (8-C), 102.28 (6-C), 127.70 (Ar–
C), 129.12 (Ar–C), 129.62 (Ar–C), 139.08 (Ar–C), 155.29 (C]O);
HRMS (ESI): calcd for
found: 396.0835.
NMR (CD3OD)
d
: 2.04 (dd, 1H, J¼8.8, 5.2 Hz, 5-H), 2.51 (t, 1H,
J¼12.4 Hz, 5-H), 2.66 (s, 3H, NCH3), 2.94 (s, 6H, NCH3), 3.26 (d,
1H, J¼9.6 Hz, 11-H), 3.40 (t, 1H, J¼9.6 Hz, 11-H), 3.69–3.87 (m,
4H, 10-H, 9-H, 8-H, 7-H), 4.68 (dd, 1H, J¼12.0, 4.8 Hz, 4-H), 6.79
(d, 2H, J¼8.8 Hz, ArH), 7.15 (d, 2H, J¼8.8 Hz, ArH); 13C NMR
C
16H20ClNO7Na ([MþNa]þ): 396.0826,
3.4.11. Compound 9e
(CD3OD) d: 33.34 (NCH3), 37.18 (5-C), 39.88 (NCH3), 57.09 (4-C),
22
White solid, mp: 242–244 ꢀC; [
a
]
þ45.41 (c 0.6, H2O); 1H NMR
61.40 (11-C), 70.34 (10-C), 73.64 (9-C), 74.48 (7-C), 75.34 (8-C),
101.25 (6-C), 113.22 (Ar–C), 127.21 (Ar–C), 128.06 (Ar–C), 151.30
D
(D2O)
d
: 2.12–2.19 (m, 1H, 5-H), 2.37 (t, 1H, J¼12.8 Hz, 5-H), 2.60 (s,
3H, NCH3), 3.23–3.54 (m, 3H, 11-H, 10-H), 3.59–3.82 (m, 3H, 7-H, 9-
(Ar–C), 154.58 (C]O); HRMS (ESI): calcd for
C18H27N2O7
H, 8-H), 4.74–4.77 (m, 1H, 4-H), 7.24–7.40 (m, 4H, ArH); 13C NMR
([MþH]þ): 383.1818, found: 383.1822.
(D2O) d: 34.52 (NCH3), 36.77 (5-C), 57.74 (4-C), 60.77 (11-C), 69.64
(10-C), 73.24 (9-C), 73.72 (7-C), 74.79 (8-C), 101.52 (6-C), 127.18 (Ar–
C), 127.76 (Ar–C), 128.94 (Ar–C), 129.71 (Ar–C), 139.58 (Ar–C),
155.58 (C]O); HRMS (ESI): calcd for C16H20ClNO7Na ([MþNa]þ):
396.0826, found: 396.0831.
3.4.17. Compound 11h
25
Pale yellow syrup; [
a]
þ2.53 (c 3.5, CHCl3); 1H NMR (CDCl3)
D
d: 2.01–2.08 (m, 1H, 2-H), 2.29–2.37 (m, 1H, 2-H), 2.43–2.51 (m,
1H, 1-H), 2.69–2.74 (m, 1H, 1-H), 2.79 (s, 3H, NCH3), 3.61 (d, 1H,
J¼9.6 Hz, 8-H), 3.72–3.89 (m, 4H, 8-H, 6-H, 4-H, 7-H), 4.18 (t, 1H,
J¼9.4 Hz, 5-H), 4.59–4.68 (m, 3H, CH2Ph), 4.76 (d, 1H, J¼12.0 Hz,
CH2Ph), 4.89–5.05 (m, 4H, CH2Ph), 6.96 (d, 2H, J¼5.6 Hz, ArH),
7.24–7.39 (m, 20H, ArH), 8.45 (d, 2H, J¼5.6 Hz, ArH); 13C NMR
3.4.12. Compound 10e
23
White solid, mp: 211–213 ꢀC; [
a
]
ꢁ20.32 (c 0.4, H2O); 1H NMR
D
(CD3OD) d: 2.32–2.39 (m, 1H, 5-H), 2.44–2.52 (m, 1H, 5-H), 2.67 (s,
3H, NCH3), 3.37–3.42 (m, 1H, 11-H), 3.46–3.53 (m, 2H, 10-H, 11-H),
3.57–3.61 (m, 1H, 7-H), 3.68–3.73 (m, 1H, 9-H), 3.96–3.99 (m, 1H, 8-
H), 4.58 (dd, 1H, J¼4.8, 12.0 Hz, 4-H), 7.34–7.48 (m, 4H, ArH); 13C
(CDCl3) d: 29.79 (1-C), 33.15 (2-C), 40.54 (NCH3), 69.21 (8-C),
72.90 (CH2Ph), 73.85 (7-C), 75.61 (CH2Ph), 75.81 (CH2Ph), 75.97
(CH2Ph), 79.16 (6-C), 80.30 (4-C), 83.95 (5-C), 104.57 (3-C),
124.30 (Ar–C), 128.03 (Ar–C), 128.11 (Ar–C), 128.27 (Ar–C), 128.44
(Ar–C), 128.75 (Ar–C), 128.81 (Ar–C), 128.92 (Ar–C), 128.96 (Ar–C),
138.51 (Ar–C), 138.84 (Ar–C), 139.02 (Ar–C), 149.93 (Py–C), 151.65
(Py–C); HRMS (ESI): calcd for C42H46N2O6 (Mþ): 674.3356, found:
674.3361.
NMR (CD3OD) d: 31.11 (NCH3), 33.72 (5-C), 51.81 (4-C), 61.96 (11-C),
70.59 (10-C), 74.57 (9-C), 75.17 (7-C), 76.54 (8-C), 102.16 (6-C),
127.32 (Ar–C), 128.63 (Ar–C), 129.29 (Ar–C), 139.76 (Ar–C), 154.73
(C]O); HRMS (ESI): calcd for C16H21ClNO7 ([MþH]þ): 374.1007,
found: 374.1012.
3.4.13. Compound 9f
3.4.18. Compound 12h
rColorless syrup; [
d: 1.86–1.94 (m, 2H, 2-H), 2.56–2.67 (m, 1H, 1-H), 2.69–2.80 (m,
23
23
White solid, mp: 238–240 ꢀC; [
a
]
D
þ36.85 (c 0.5, H2O); 1H NMR
a]
þ5.64 (c 1.7, CHCl3); 1H NMR (CDCl3)
D
(CD3OD)
d
: 2.10 (dd, 1H, J¼14.0, 5.2 Hz, 5-H), 2.49 (t, 1H, J¼12.4 Hz,
5-H), 2.71 (s, 3H, NCH3), 3.28–3.42 (m, 2H, 11-H), 3.71–3.88 (m, 4H,
10-H, 9-H, 8-H, 7-H), 4.72–4.76 (m, 1H, 4-H), 5.98 (s, 2H, CH2), 6.83–
1H, 1-H), 3.44 (d, 1H, J¼9.2 Hz, 8-H), 3.68–3.77 (m, 3H, 8-H, 6-H,
4-H), 4.00–4.03 (m, 2H, 7-H, 5-H), 4.53–4.69 (m, 4H, CH2Ph),
4.84–4.97 (m, 4H, CH2Ph), 7.00 (d, 2H, J¼5.6 Hz, ArH), 7.20–7.36
6.86 (m, 3H, ArH); 13C NMR (CD3OD)
d: 33.48 (NCH3), 37.26 (5-C),
57.49 (4-C), 60.56 (11-C), 70.34 (10-C), 73.63 (9-C), 74.40 (7-C),
75.40 (8-C), 101.24 (6-C), 101.75 (CH2), 106.99 (Ar–C), 108.50 (Ar–C),
121.09 (Ar–C), 133.80 (Ar–C), 148.21 (Ar–C), 148.41 (Ar–C), 154.47
(C]O); HRMS (ESI): calcd for C17H21NO9Na ([MþNa]þ): 406.1114,
found: 406.1119.
(m, 20H, ArH), 8.44 (d, 2H, J¼5.6 Hz, ArH); 13C NMR (CDCl3)
d:
28.45 (1-C), 39.34 (2-C), 69.17 (8-C), 72.08 (CH2Ph), 73.77 (7-C),
75.33 (CH2Ph), 75.86 (CH2Ph), 76.05 (CH2Ph), 78.73 (6-C), 82.10
(4-C), 84.33 (5-C), 98.18 (3-C), 124.29 (Ar–C), 128.07 (Ar–C),
128.15 (Ar–C), 128.26 (Ar–C), 128.50 (Ar–C), 128.77 (Ar–C), 128.83
(Ar–C), 128.88 (Ar–C), 128.93 (Ar–C), 138.13 (Ar–C), 138.62 (Ar–C),
138.71 (Ar–C), 138.91 (Ar–C), 149.92 (Py–C), 151.71 (Py–C); HRMS
3.4.14. Compound 10f
22
White solid, mp: 178–180 ꢀC; [
a]
ꢁ18.56 (c 0.5, H2O); 1H
(ESI): calcd for
668.2996.
C
41H43NO6Na ([MþNa]þ): 668.2988, found:
D
NMR (CD3OD)
d: 2.37–2.48 (m, 2H, 5-H), 2.72 (s, 3H, NCH3),
3.33–3.54 (m, 4H, 11-H, 10-H, 9-H), 3.69–3.73 (m, 1H, 7-H), 4.10–
4.15 (m, 1H, 8-H), 4.51–4.54 (m, 1H, 4-H), 5.99 (s, 2H, CH2), 6.86
3.4.19. Compound 12i
Pale yellow syrup; [
23
(d, 3H, ArH); 13C NMR (CD3OD)
d: 30.98 (NCH3), 33.37 (5-C),
a]
þ10.20 (c 1.1, CHCl3); 1H NMR (CDCl3)
d:
D
56.90 (4-C), 62.02 (11-C), 70.59 (10-C), 74.64 (9-C), 75.19 (7-C),
76.62 (8-C), 101.75 (CH2), 102.07 (6-C), 107.06 (Ar–C), 108.49 (Ar–
C), 121.10 (Ar–C), 133.54 (Ar–C), 148.25 (Ar–C), 148.90 (Ar–C),
154.61 (C]O); HRMS (ESI): calcd for C17H21NO9Na ([MþNa]þ):
406.1114, found: 406.1121.
1.30–1.33 (m, 1H, 2-H), 1.46–1.49 (m, 1H, 2-H), 2.06–2.16 (m, 1H,
1-H), 2.43–2.47 (m, 1H, 1-H), 3.34 (d, 1H, J¼9.6 Hz, 8-H), 3.71–3.81
(m, 3H, 8-H, 6-H, 4-H), 4.13–4.15 (m, 5-H), 4.34–4.36 (m, 7-H), 4.56–
4.65 (m, 4H, CH2Ph), 4.77 (d, 1H, J¼12.0 Hz, CH2Ph), 4.85–5.05 (m,
3H, CH2Ph), 7.06–7.39 (m, 22H, ArH), 7.58–7.67 (m, 1H, ArH), 8.56–
8.59 (m,1H, ArH); 13C NMR (CDCl3)
d: 30.09 (1-C), 34.66 (2-C), 69.30
3.4.15. Compound 9g
(8-C), 71.33 (CH2Ph), 73.93 (7-C), 75.34 (CH2Ph), 76.13 (CH2Ph),
76.32 (CH2Ph), 79.08 (6-C), 80.19 (4-C), 83.63 (5-C), 98.75 (3-C),
122.42 (Ar–C), 122.60 (Ar–C), 127.90 (Ar–C), 127.97 (Ar–C), 128.13
(Ar–C), 128.23 (Ar–C), 128.37 (Ar–C), 128.55 (Ar–C), 128.80 (Ar–C),
128.98 (Ar–C), 136.74 (Ar–C), 138.60 (Ar–C), 149.57 (Py–C), 161.72
(Py–C); HRMS (ESI): calcd for C41H44NO6 ([MþH]þ): 646.3169,
found: 646.3181.
21
White solid, mp: 216–218 ꢀC; [
a
]
ꢁ20.52 (c 0.3, H2O); 1H NMR
D
(CD3OD)
d
: 2.06 (dd, 1H, J¼8.8, 5.2 Hz, 5-H), 2.53 (t, 1H, J¼12.4 Hz,
5-H), 2.68 (s, 3H, NCH3), 2.95 (s, 6H, NCH3), 3.37–3.45 (m, 2H, 11-H),
3.73–3.89 (m, 4H, 10-H, 9-H, 8-H, 7-H), 4.70 (dd, 1H, J¼12.0, 4.8 Hz,
4-H), 6.81 (d, 2H, J¼8.8 Hz, ArH), 7.17 (d, 2H, J¼8.8 Hz, ArH); 13C
NMR (CD3OD) d: 33.39 (NCH3), 37.20 (5-C), 40.00 (NCH3), 57.10

X. Li et al. / Tetrahedron 64 (2008) 9911–9920
9919
3.5. X-ray crystallographic measurement of single
crystal of 9a1
a 5% CO2 humidified incubator. Compounds were added to the wells
at final concentrations of 1, 10, and 100 mol/L. Control wells were
m
prepared by addition of DMEM. Wells containing DMEM without
cells were used as blanks. The plates were incubated at 37 ꢀC in a 5%
CO2 incubator for 48 h. Upon completion of the incubation, stock
The single crystal of the debenzylated compound 9a1 was
obtained by recrystallization from the solution of H2O/EtOH and
applied on a Bruker SMART CCD diffractometer for analysis.
The intensity data were collected on a Bruker SMART CCD dif-
MTT dye solution (10
mL, 5 mg/mL) was added to each well. After
4 h incubation, the supernatant was removed and DMSO (100
mL)
fractometer with graphite-monochromated Mo
K
a
radiation
was added to solubilize the MTT. The optical density of each well
was measured on a microplate spectrophotometer at a wavelength
of 570 nm. The inhibition rate was calculated according to the
formula: (ODcontrolꢁODtreated)/ODcontrolꢂ100%.
(
l
¼0.71073 Å) using the /2 scan technique from a single-crystal
q u
of 0.30 mmꢂ0.20 mmꢂ0.20 mm, and a semi-empirical absorption
correction was applied for all complexes. The structures were
solved by direct methods and refined by full-matrix least-squares
on F2. The absolute structure parameter was ꢁ0.5 (13). All non-
hydrogen atoms were refined anisotropically. The crystallographic
structure is shown in Figure 1.
Acknowledgements
The financial supports from the National Natural Science
Foundations of China (NSFC) (20472015 and 20672027), the Natural
Science Foundations of Hebei (2005000106 and B2008000588),
and the Research Foundation from the Ministry of Education of
China (206013) are gratefully acknowledged.
3.6. Biological activity assay
3.6.1. Glycosidases inhibition
Inhibitory activity of the synthesized compounds against
lase, -glucosidase, and -glucosidase: The enzymes -glucosidase
(yeast) and -glucosidase (almonds) were obtained from Fluka;
-amylase (Bacillaceae, 4000u/mg)fromSanland-chenInternational
Inc., Xiamen, China. Two substrates p-nitrophenyl -glucopyrano-
side (PNPG) and
a-amy-
a
b
a
References and notes
b
a
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a
D
-(ꢁ) salicin were purchased from Sigma Chemical
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Table 4.
´
´
lan˜os, J. G.; Lopez, O. Heterocycles from Carbohydrate Isothiocyanates. In Topics
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a
-Amylase assay was performed using 1% starch as substrate
in pH¼6.0 phosphate buffers at 50 ꢀC. Enzyme solution (0.1 mL,
5 mg of solid enzyme in 50 mL of pH¼6.0 phosphate buffer),
0.1 mL of inhibitor (1 mg/mL), and 1 mL of buffer were incubated
for 10 min, and then 1 mL of substrate was added. After 10 min,
2 mL of 3,5-dinitrosalicylic acid was added and then the reaction
was heated in boiling water for 5 min. The solution was diluted
to 20 mL after cooling down. Absorbance readings were taken
on
a TU-1901 UV–vis spectrophotometer at 540 nm using
distilled deionized water as a blank control and acarbose as
a positive control.
b
-Glucosidase assay was performed using
D
-(ꢁ) salicin (2 mg/
mL) as substrate in pH¼4.8 phosphate buffers at 35 ꢀC. Enzyme
solution (0.1 mL, 10 mg of solid enzyme in 10 mL of pH¼4.8 acetate
buffer), 0.1 mL of inhibitor (1 mg/mL), and 0.9 mL of buffer were
incubated for 10 min, and then 0.8 mL of substrate was added. After
10 min, 2 mL of 3,5-dinitrosalicylic acid was added and then the
reaction mixture was heated in boiling water for 5 min. The solu-
tion was diluted to 20 mL after cooling down. Absorbance readings
were taken on a TU-1901 UV–vis spectrophotometer at 540 nm
using distilled deionized water as a blank control.
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a
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substrate in pH¼6.8 phosphate buffers at 37 ꢀC. Enzyme solution
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buffer), 0.1 mL of inhibitor (1 mg/mL), 1.9 mL of buffer, and
0.05 mL glutathione (reduced, 1 mg/mL) were incubated for
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was quenched with 10 mL sodium carbonate (0.1 mol/L) after
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down. Absorbance readings were taken on a TU-1901 UV–vis
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3.6.2. Antitumor activity
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man cervical cancer cells) was examined by the modified Mos-
mann’s protocol as follows. Briefly, cells (104 cells per well) were
plated in 96-well culture plates and cultured overnight at 37 ꢀC in
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12. CCDC 694485 contains the supplementary crystallographic data for this paper.
These data can be obtained free of charge via www.ccdc.cam.ac.uk/data_re-
quest/cif, or by emailing data_request@ccdc.cam.ac.uk, or by contacting The
Cambridge Crystallographic Data Centre, 12, Union Road, Cambridge CB2 1EZ,
UK; fax: þ44 1223 336033.
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