A convenient synthesis of novel pyranosyl homo-C-nucleosides and their antidiabetic activities

Article abstract of DOI:10.1016/j.carres.2011.03.006

A series of pyranosyl homo-C-nucleosides have been synthesized by reaction of butenonyl C-glycosides (5a-5j, and 8) and cyanoacetamide in presence of t-BuOK followed by further modifications. The reaction proceeds by Michael addition of cyanoacetamide to

Full text of DOI:10.1016/j.carres.2011.03.006

Carbohydrate Research 346 (2011) 1191–1201
Contents lists available at ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
A convenient synthesis of novel pyranosyl homo-C-nucleosides
and their antidiabetic activities
Surendra Singh Bisht a, Natasha Jaiswal b, Anindra Sharma a, Seerat Fatima a, Rahul Sharma a, Neha Rahuja b,
A. K. Srivastava b, Vikas Bajpai c, Brijesh Kumar c, Rama P. Tripathi a,
⇑
a Medicinal and Process Chemistry Division, Central Drug Research Institute, Lucknow 226 001, CSIR, India
b Biochemistry Division, Central Drug Research Institute, Lucknow 226 001, CSIR, India
c Sophisticated Advanced Instrumentation Facility Division, Central Drug Research Institute, Lucknow 226 001, CSIR, India
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of pyranosyl homo-C-nucleosides have been synthesized by reaction of butenonyl C-glycosides
(5a–5j, and 8) and cyanoacetamide in presence of t-BuOK followed by further modifications. The reaction
proceeds by Michael addition of cyanoacetamide to the butenonyl C-glycosides and subsequent dehyd-
rative cyclization and oxidative aromatization to give glycosylmethyl pyridones (6a–6j, 7a–7j, 9, and
10). The glycosylmethyl pyridones (6a–6e) on reaction with POCl3 under reflux gave respective
glycosylmethyl pyridines (11a–11e and 12a–12e) in good yields. The synthesized compounds were
Received 31 December 2010
Received in revised form 15 February 2011
Accepted 2 March 2011
Available online 8 March 2011
Keywords:
screened for their in vitro a-glucosidase, glucose-6-phosphatase and glycogen phosphorylase inhibitory
Homo-C-nucleoside
Glycosylmethyl pyridones
Glycosylmethyl pyridines
activities. One of the pyridylmethyl homo-C-nucleoside, compound 11d, displayed 52% inhibition of glu-
cose-6-phosphatase as compared to the standard drug sodium orthovanadate while compound 12a
showed a significant antihyperglycemic effect of 17.1% in the diabetic rats as compared to the standard
drug metformin.
a-Glucosidase
Glucose-6-phosphatase
Glycogen phosphorylase
Ó 2011 Elsevier Ltd. All rights reserved.
1. Introduction
>90% of renal glucose re-absorption.12,13 Selective inhibition of
SGLT2, therefore, results in preventing glucose re-absorption,
The synthetic interest in nucleoside analogs is due to their capa-
bility to interfere with functioning of enzymes, which renders
them antiviral, anticancer, antibacterial, insecticidal, fungicidal,
and herbicidal, among other known biological activities. The de-
mand and utility of nucleosides and their analogs is continuously
increasing due to the spread of HIV, hepatitis B, herpes simplex
and other viruses.1–6 The homo-C-nucleosides, which have a
–CH2– linker between the sugar and heterocyclic moiety are also
of great biological significance and are potent chemotherapeutic
which may lead to new antidiabetic drugs with no gastrointestinal
side effects. Nucleoside analogues and pyridine-based compounds
are known to inhibit the above glycosidases and result in limiting
hyperglycemia. Several aryl C-glycosides (dapagliflozin, sergliflo-
zin, LX-4211) and many other heteroaromatic-C-glycosides such
as compound 4 are potent SGLT2 inhibitors13–15 (Fig. 1) and pos-
sess very good antidiabetic activities in vivo. The dapagliflozin is
in phase III clinical trial for the treatment of diabetes.14 Further,
certain nucleosides and pyridine derivatives are known to inhibit
hepatic glycosidases and possess a hypoglycemic effect.16,17
A number of methods exist in literature toward the synthesis of
aryl b-C-glycosides18–21 including those clinical candidates for the
treatment of diabetes. In general, methods known so far, are based
on nucleophilic attack at the anomeric carbon of the sugar, the gly-
cosylidene carbene formation,22–24 or Wittig-type reaction25 or
substitution of the hydroxyl group at the anomeric carbon with
halogen and subsequent reactions such as Reformatsky,26 Grig-
nard,27 or free-radical reactions.28 The methods adopted for the
synthesis of antidiabetic aryl b-C-glycosides involve the use of BuLi
and restricted reaction conditions. Most of these methods of b-C-
glycoside syntheses require a metal-based catalyst and inert atmo-
sphere for successful completion of the reaction. Keeping in mind
the above facts and in continuation of our ongoing studies toward
the synthesis of b-C-glycosides29–31 as new antidiabetic agents, we
agents or enzyme inhibitors.7,8 Further, among the
a- and b-ano-
meric nucleosides, the latter have extensively been studied due
to their natural occurrence and numerous biological activities,
while the a
-nucleosides have received little attention.9–11
Diabetes mellitus commonly known as diabetes is a group of
metabolic diseases characterized by abnormally high levels of plas-
ma glucose. Inhibition of glycosidases, glucose-6-phosphatase and
glycogen phosphorylase are known to reduce the hyperglycemia.
Very recently renal SGLT2 (sodium-dependant glucose co-trans-
porter), located on the surface of the epithelial cells and lining
the S1 segment of the proximal tubule, is reported to facilitate
⇑
Corresponding author. Tel.: +91 0522 2612411; fax: +91 522 2623405/
2623938/2629504.
E-mail addresses: rpt.cdri@gmail.com, rp_tripathi@cdri.res.in (R.P. Tripathi).
0008-6215/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2011.03.006

1192
S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
OR
Methylene linker
O
OR'
Cl
HO
HO
Et
OH
OR
O
Methylene linker
N
Cl
1
. R=H, R'=-Et, dapagliflozin
HO
HO
2
, R=-COOEt, R'=-CH3, Sergliflozin
N
OH
3, R=CH3, R'=Et, LX-4211
Cl
4
OH
O
HO
HO
R'
N
OH
R''
Methylene linker
Proposed molecules
Figure 1. Antidiabetic b-C-glucosides 1–4 in clinical trial and our designed molecules.
were interested in the synthesis of heteroaryl b-C-glycosides as
homo-C-nucleoside analogs and evaluating their antidibetic
activities.
(1–10 equiv) in different organic solvents, under the influence of
various inorganic and organic bases, and under an N2 atmosphere
at ambient temperature for 30 min (until the disappearance of
starting butenoyl C-glycoside) was carried out. The reaction mix-
ture was brought under the influence of an O2 atmosphere to carry
out oxidative aromatization. The reaction mixture was subse-
quently acetylated with Ac2O/pyridine (Scheme 1, Table 1) in order
to facilitate the isolation of the pure product by column chroma-
tography. Four equivalents of t-BuOK in DMSO at ambient temper-
ature proved to be the most optimum reaction conditions (entry 6)
followed by in situ acetylation of the reaction mixture to give the
desired compound 3-cyano-4-(40-fluorophenyl)-6-[(200,300,400,600-tet-
Our method of pyranosyl homo-b-C-nucleoside synthesis in-
volves reaction of the butenonyl C-glycosides with cyanoacet-
amide followed by further modification of the intermediates. The
reaction proceeds by 1,4-conjugate addition (Michael addition) of
cyanoacetamide on to the butenonyl C-glycoside to give an inter-
mediate which on dehydrative cyclization and subsequent oxida-
tive aromatization offers the glycopyranosyl methyl pyridine. The
latter on refluxing with POCl3 gives the respective pyridylmethyl
glycosides. The compounds are evaluated for their antidiabetic po-
tential in vitro. The method adopted in the synthesis is quite sim-
ple, eco-friendly, and economical as it involves easily available
sugars and reagents.
ra-O-acetyl-b-D-glucopyranosyl)methyl]pyridone (6a) in 53% yield
(Table 1). It is appropriate to mention here that many other minor
products observed on TLC plate could not be isolated in pure forms
despite our several attempts.
The structure of 3-cyano-4-(40-fluorophenyl)-6-[(200,300,400,600-
2. Results and discussion
tetra-O-acetyl-b-
D-glucopyranosyl)methyl]pyridone
(6a)
was
established on the basis of its spectroscopic data and analysis.
2.1. Chemistry
The IR spectrum exhibited absorption bands at 3467 cmꢀ1
,
1662 cmꢀ1 and 1753 cmꢀ1 indicating the presence of NH and N–
C@O groups of the pyridone ring and carbonyl of the sugar acetyl
groups, while an absorption band at 2227 cmꢀ1 indicated the pres-
ence of cyano (CN) group. ESIMS of the compound displayed m/z
559 [M+H]+ peak. In the 1H NMR spectrum the exchangeable
The starting butenonyl C-glycosides (5a–5j) were prepared
from commercially available
D-glucose following our method and
earlier reported protocols.29–32 To optimize the reaction condi-
tions, the reaction of 1 equivalent of (E)-1-(b-D-glucopyranosyl)-
4-(40-fluorophenyl)-but-3-en-2-one (5a) with cyanoacetamide
OH
OH
F
F
O
(i) Cyanoacetamide,
O
HO
HO
base (1-10 eq.),
HO
HO
OH
solvents, N2, O2,
25 °C, 3-10 h
OH
O
HN
CN
O
(5a)
(ii) in situ,Ac2O, pyridine
OH
OAc
F
F
O
O
(iii) NaOMe/MeOH
(20 mol %), 1 h
HO
HO
AcO
AcO
OH
OAc
HN
HN
CN
CN
O
O
(6a)
(7a)
Scheme 1. Optimization of reaction conditions with compound 5a and cyanoacetamide.

S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
1193
Table 1
carbons OCH2 and CH2 were observed at d 62.4 and d 39.9. The four
methyl carbons of the acetyl groups were observed at d 20.8, 20.7,
20.6, and 20.5.
Once the reaction conditions were optimized, the scope of
this reaction was studied with different substrates. Thus the
Optimization of reaction between (E)-1-(b-D-glucopyranosyl)-4-(40-fluorophenyl)but-
3-en-2-one (5a) and cyanoacetamide
Entry Base
Solvent Reaction time
(h)
Isolated yield (%)
1
2
t-BuOKa
THF
DMF
10
10
NRb
reaction of (E)-1-(b-D-glucopyranosyl)-4-(aryl)but-3-en-2-ones
t-BuOKa
Complex reaction
(5b–5j) separately with cyanoacetamide was carried out
(Scheme 2) under the above optimized reaction conditions to
give the respective 3-cyano-4-(aryl)-6-[(200,300,400,600-tetra-O-acet-
yl-b-D-glucopyranosyl)methyl]pyridones (6b–6j) in moderate to
good yields (Scheme 2). Subsequently, deacetylation of
mixturec
3
4
5
6
7
8
9
10
t-BuOKa
t-BuOKa
NaOMea
t-BuOKa
Et3Na
MeOH
CH2Cl2
MeOH
DMSO
DMSO
DMSO
DMSO
DMSO
10
10
10
3
10
10
10
10
20
NRb
15
53
NRb
compounds 6b–6j with NaOMe/MeOH afforded the respective
DBU
NRb
3-cyano-4-phenyl-6-[(b-
D
-glucopyranosyl)
methyl]pyridones
NRb
a
Cs2CO3
LiOHa
Complex reaction
(7b–7j) in good yields. The structures of all these compounds
were in accordance with their spectroscopic data.
mixturec
a
11
12
K2CO3
DMSO
DMSO
10
10
5
10
Further, in order to enhance the scope of this reaction, a
t-BuOK
disaccharide
derivative,
(E)-1-(20,30,60,200,300,400,600-hepta-O-
(1 equiv)
t-BuOK
(3 equiv)
t-BuOK
acetyl-b-cellobiosyl)-4-(2-napthyl)but-3-en-2-one
(8),29
was
13
14
DMSO
DMSO
10
3
25
50
reacted with cyanoacetamide to give the respective
3-cyano-4-(20-napthyl)-6-[(200,300,600,2000,3000,4000,6000-hepta-O-acetyl-b-
cellobiosyl)methyl]pyridone (9) in 54% yield. The deacetylation of
the acetyl groups in compound 9 afforded to 3-cyano-4-(20-nap-
thyl)-6-[(b-cellobiosyl)methyl] pyridone (10) in 76% yield
(Scheme 3).
(10 equiv)
a
b
c
4 equiv.
NR = No reaction.
Reaction mixture showed a streak on the TLC plate and no desired product was
detected.
Finally, in order to create structural diversity in the heteroaro-
matic ring, a few selected 3-cyano-4-(aryl)-6-[(200,300,400,600-tetra-
O-acetyl-b-
D
-glucopyranosyl)methyl]pyridones
(6a–6e)
were
proton (NH) and H-5 of pyridone ring were observed at d 12.5 (br s)
and 6.38 (s), respectively, while the aromatic protons were ob-
served as two multiplets in the range of d 7.73–7.68 and d 7.44–
7.38. The H-200, H-300 and H-400 sugar ring appeared as a three dis-
reacted with phosphorous oxychloride (POCl3) at 100 °C to afford
the respective 2-chloro-3-cyano-4-(aryl)-6-[(200,300,400,600-tetra-O-
acetyl-b-D-glucopyranosyl)methyl] pyridines (11a–11e) in good
yields (Scheme 4). Deacetylation of the above compounds (11a–
11e) with NaOMe/MeOH gave respective 2-chloro-3-cyano-4-
tinct triplets at
d 5.23 (t, 1H, J = 9.45 Hz), d 4.87 (t, 1H,
J = 9.72 Hz), d 4.79 (t, 1H, J = 9.57 Hz), respectively. The H-100 and
H-6a00 of the sugar moiety were observed as multiplet in the range
of d 4.12–4.05 while H-6b00 and H-500 were visible as multiplet in
the range of d 3.97–3.91. The protons of the four sugar acetyl
groups were visible as singlets at d 2.03, d 2.02, d 2.01, and d
1.98. In 13C NMR spectrum the signals for the acetyl carbonyl car-
bons appeared at d 170.5, 170.2, 170.1, and 169.9 while C„N car-
bon signal was visible at d 116.2. The aromatic and heteroaromatic
carbons were observed in the range of d 161.7–99.9, while the C-100,
C-200, C-300, C-400, and C-500 of the puranose sugar appeared at d 74.7,
73.7, 72.7, 71.7, and d 68.7, respectively. The two methylene
(aryl)-6-[(b-
D-glucopyranosyl)methyl]pyridines
(12a–12e)
in
quantitative yields (Scheme 4). The structures of all these com-
pounds were in full agreement with their spectroscopic data.
2.2. Biology
The above-synthesized compounds were screened against the
a
-glucosidase enzyme, glucose-6-phosphatase and glycogen phos-
phorylase.33–36 The compounds dissolved in DMSO were evaluated
for their inhibitory potential against the above three enzymes at a
OAc
OH
O
(i) Cyanoacetamide, DMSO
t-BuOK, N2, O2, 25 °C, 2-3h
O
AcO
AcO
HO
HO
Ar
Ar
OAc
OH
(ii) Pyridine, Acetic anhydride
0 °C to 25 °C, 1-2h
HN
O
CN
O
(5b-5j)
(6b-6j)
5b,6b,7b
: Ar = 4-bromophenyl
NaOMe/MeOH
(20 mol%),
25 °C, 1-2h
5c,6c,7c: Ar = 4-benzyloxyphenyl
5d,6d,7d: Ar = 4-methoxyphenyl
5e,6e,7e: Ar = 3,4-dimethoxyphenyl
5f,6f,7f: Ar = 4-chlorophenyl
5g,6g,7g: Ar = phenyl
5h,6h,7h: Ar = 3,4,5-trimethoxyphenyl
OH
O
HO
HO
5i,6i,7i
5j,6j,7j
; Ar = 2-napthyl
: Ar = 4-O-acetylphenyl
Ar
CN
OH
HN
O
(7b-7j)
Scheme 2. Synthesis of glycosylmethylpyridones 6b–6j and 7b–7j.

1194
S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
OH
OH
OAc
OAc
O
(i) Cyanoacetamide, DMSO,
t-BuOK, N2, O2, 25 °C, 2-3h
O
O
O
HO
HO
O
HO
O
AcO
AcO
AcO
OH
OH
(ii) Pyridine, acetican hydride,
OAc
OAc
O
0 °C to 25 °C, 1-2h
HN
CN
(9)
(8)
O
NaOMe/MeOH
(20 mol%)
25 °C, 2h
OH
OH
O
O
O
HO
HO
HO
OH
OH
HN
CN
(10)
O
Scheme 3. Synthesis of cellobiosylmethylpyridones 9 and 10.
OAc
O
OAc
O
POCl3, 100 °C, 1h
AcO
AcO
AcO
AcO
Ar
Ar
OAc
OAc
N
HN
CN
CN
O
Cl
(11a-11e)
(6a-6e)
NaOMe/MeOH
(20 mol%)
25 °C, 1-2h
OH
O
6a,11a,12a: Ar = 4-fluorophenyl
6b,11b,12b: Ar = 4-bromophenyl
6c,11c,12c: Ar = 4-benzyloxyphenyl
6d,11d,12d: Ar = 4-methoxyphenyl
6e,11e,12e: Ar = 3,4-dimethoxyphenyl
HO
HO
Ar
CN
OH
N
Cl
(12a-12e)
Scheme 4. Synthesis of glycosylmethylpyridines 11a–11e and 12a–12e.
concentration of 100 lM. Percentage inhibition was calculated
3. Conclusions
with respect to control. The enzyme inhibitory data of screened
compounds are shown in Table 2.
In conclusion, we have developed a quite simple and economi-
cal method for the synthesis of pyranosyl homo-C-nucleosides
exclusively with the b-configuration. The method basically in-
volves Michael addition, dehydrative cyclization and oxidative aro-
matization reactions. A few of the compounds displayed moderate
As evident from activity results (Table 2) among all the com-
pounds screened, compound 11d with a 4-methoxyphenyl substi-
tuent exhibits good inhibitory activity (52.2% inhibition), while
compounds 6h, 6c, 7f, and 12c display inhibition in the range of
43.3–46% against glucose-6-phosphatase. The compounds 11b
and 12b exhibited moderate inhibitory activity with 41.3% and
41.8% inhibition against glycogen phosphorylase, respectively.
Out of all the compounds screened above, a few exhibited mod-
erate inhibitory potential for the three enzymes as compared to
standard control. Out of curiosity one of the compounds 12a, with
a 4-flurophenyl substituent inhibiting all the three enzymes
in vitro was selected for in vivo evaluation in a streptozotocin
(STZ) induced diabetic rat model. The in vivo results are shown
in Table 3 and Figure 2. It is evident that compound 12a caused a
significant decline by 16.3% (p <0.05 in 5 h) and 17.1% (p <0.05 in
24 h), respectively, in the hyperglycemia of the post sucrose-
loaded diabetic rats at 100 mg/kg. The standard drug metformin
at the same dose level (100 mg/kg) showed a blood glucose lower-
ing effect of 26.9% (p <0.01 after 5 h) and 21.4% (p <0.01 after 24 h),
respectively. The effect of compound 12a, the standard drug met-
formin, and a control (no drug) at different time intervals is also
shown in Figure 2.
inhibition of either of
a-glucosidase, glucose-6-phosphatase, or
glycogen phosphorylase. However, one of the compounds showed
antihyperglycemic effect in STZ-induced diabetic rats. These com-
pounds are being selectively exploited for diversification to create
a new class of compounds with chemotherapeutic potential.
4. Experimental section
4.1. Chemistry
Commercially available reagent grade chemicals were used
as received. All reactions were monitored by TLC on E. Merck
Kieselgel 60 F254 with detection by UV light or by spraying a 20%
KMnO4 aq solution and/or spraying a 4% H2SO4 ethanolic solution
followed by heating. Column chromatography was performed on
silica gel (60–120 mesh E. Merck). IR spectra were recorded as
thin films or in KBr solution with a Perkin–Elmer Spectrum RX-1
)
(4000–450 cmꢀ1 spectrophotometer. The 1H (200 MHz and

S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
1195
Table 2
-Glucosidase, glucose-6-phosphatase, and glycogen phosphorylase inhibitory activ-
ity of synthesized compounds
Control
12 a
Metformin
a
600
S.
No.
Compounds
% Activity (100
l
M)
500
400
300
200
100
0
a
-
Glucose-6-
phosphatase
inhibition
Glycogen
phosphorylase
inhibition
Glucosidase
inhibition
1
2
3
4
5
6
7
8
9
6a
6b
6c
6d
6e
6f
6g
6h
6i
6j
7a
7b
7c
7d
7e
7f
7g
7h
7i
7j
9
10
11a
11b
11c
11d
11e
12a
12b
12c
12d
12e
Acarbose
Sodium-o-
vandate
+11.4
ꢀ13.0
+15.6
ꢀ8.33
ꢀ13.7
ꢀ7.50
+29.0
+12.5
+3.30
ꢀ8.6
ꢀ11.9
ꢀ13.7
ꢀ45.0
+19.1
+7.4
ꢀ6.38
ꢀ19.4
ꢀ46.0
ꢀ21.3
+4.9
ꢀ26.9
ꢀ28.7
+17.7
+0.98
+6.1
ꢀ18.6
+20.6
ꢀ2.94
ꢀ36.3
+14.6
ꢀ26.1
ꢀ37.4
+8.9
ꢀ2.95
ꢀ18.6
ꢀ36.3
+1.96
ꢀ25.3
+12.8
+6.9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
+3.30
ꢀ11.2
0.0
ꢀ16.4
ꢀ21.5
+4.4
Time (min)
Figure 2. Antihyperglycemic effect of compound 12a and standard drug metformin
on the blood glucose levels of streptozotocin-induced diabetic rats post sucrose
load at various time intervals.
ꢀ25.0
ꢀ14.4
ꢀ8.40
+20.3
ꢀ4.40
ꢀ20.7
ꢀ3.3
0.0
ꢀ8.03
ꢀ42.6
ꢀ25.5
ꢀ23.4
+6.40
+5.0
4 equiv of t-BuOK (12.2 mmol) under nitrogen atmosphere at
room temperature. After stirring for 30 min, the N2 atmosphere
was replaced by an O2 atmosphere, and the reaction was further
stirred until the disappearance of starting material was con-
firmed (TLC). The reaction mixture was then brought to 0 °C
and subsequently 50 mL of pyridine was added. To the stirring
reaction mixture Ac2O (24.3 mmol) was slowly added, and stir-
ring was continued until completion of the reaction (TLC). The
reaction mixture was extracted with EtOAc/water, and the or-
ganic layer was dried (Na2SO4) and concentrated under reduced
pressure to give a crude mass. The latter was purified by column
chromatography on silica gel (60–120 mesh) using MeOH–CHCl3
as eluant to give corresponding 3-cyano-4-(aryl)-6-[(200,300,400,600-
ꢀ29.7
0.0
+5.8
+8.0
+9.9
+12.4
+16.5
ꢀ41.3
ꢀ26.9
+11.3
ꢀ32.2
ꢀ16.5
ꢀ41.8
ꢀ6.70
+2.61
+18.3
—
ꢀ30.4
ꢀ4.90
ꢀ7.43
ꢀ0.86
ꢀ9.42
ꢀ22.1
+5.43
ꢀ7.80
ꢀ9.50
ꢀ15.5
ꢀ39.0
—
+29.8
ꢀ31.3
ꢀ29.5
ꢀ52.2
ꢀ26.9
ꢀ34.3
ꢀ35.8
ꢀ43.3
ꢀ2.90
ꢀ17.9
—
ꢀ39.0
—
tetra-O-acetyl-b-D-glucopyranosyl)methyl]pyridones (6a–6j).
4.1.1.1. 3-Cyano-4-(40-fluorophenyl)-6-[(200,300,400,600-tetra-O-ace
tyl-b- -glucopyranosyl)methyl]pyridone (6a). Compound 6a
was obtained by the reaction of (E)-1-(b- -glucopyranosyl)-4-
Table 3
D
Antihyperglycemic activity of compound 12a and the standard drug metformin in
sucrose-challenged STZ-induced diabetic rats
D
(40-fluorophenyl)but-3-en-2-one 5a (0.58 g, 1.78 mmol), cyano-
acetamide (0.15 g, 1.78 mmol), t-BuOK (0.80 g, 7.14 mmol),
pyridine (10 mL), and Ac2O (1.0 mL, 10.7 mmol) as white solid
(0.52 g, yield 53%); mp >240 °C; Rf 0.5 (2:98, MeOH–CHCl3);
Compound
Dose (mg/kg)
% Antihyperglycemic activity
24 h
5 h
12a
Metformin
100
100
16.3a
26.9b
17.1a
21.4b
½
a 2D5
ꢁ
ꢀ63.2 (c 0.1, CHCl3); IR (KBr): mmax 3467 (NH stretching),
a
p <0.05.
p <0.01.
2227 (C„N stretching), 1753 (O–C@O stretching), 1662 cmꢀ1
(N–C@O stretching); 1H NMR (300 MHz, DMSO-d6): d 12.5 (br
s, 1H, NH), 7.73–7.68 (m, 2H, ArH), 7.44–7.38 (m, 2H, ArH),
6.38 (s, 1H, H-5), 5.23 (t, 1H, J = 9.45 Hz, CH), 4.87 (t, 1H,
J = 9.72 Hz, CH), 4.79 (t, 1H, J = 9.57 Hz, CH), 4.12–4.05 (m, 2H,
H-100, H-600a), 3.97–3.91 (m, 2H, H-600b, H-500), 2.91 (dd, 1H,
b
300 MHz) and 13C NMR (50 or 75.5 MHz) spectra were recorded on
a Bruker DRX-300 instrument in DMSO, CD3OD, and CDCl3. Chem-
ical shift values are reported in ppm relative to TMS (tetramethyl-
silane) as internal reference unless otherwise stated; s (singlet), d
(doublet), t (triplet), m (multiplet), dd (double doublet), ddd (dou-
blet of double of doublet), br s (broad singlet); J in hertz. Mass
spectra were taken on a JMS-100 TLC (Accutof) atmospheric pres-
sure ionization time-of-flight mass spectrometer (Jeol, Tokyo, Ja-
pan) fitted with a DART ion source operated in positive-ion
mode. Optical rotations were measured in a 1.0 dm tube with a Ru-
dolph Autopol III polarimeter in MeOH or CHCl3. Elemental analy-
ses were performed on a Perkin–Elmer 2400 II elemental analyzer.
J1 = 3.0 Hz, J2 = 15.0 Hz, –CHaH–), 2.72 (dd, 1H, J = 5.9 Hz,
CHbH–), 2.03, 2.02, 2.01, 1.98 (each singlet, 12H, 4 ꢂ –OCOCH3);
13C NMR (50 MHz, DMSO-d6):
170.5, 170.2, 170.1, 169.9
–
d
(OCOCH3), 161.7, 159.3, 152.0, 131.0 (ArCH), 130.9 (ArCH),
116.8 (ArCH), 116.5 (ArCH), 116.2 (C„N), 107.5, 99.9, 74.7,
73.7, 72.7, 71.7, 68.7, 62.4 (OCH2), 39.9 (CH2), 20.8, 20.7, 20.6,
20.5 (CH3); ESIMS: m/z 559 (M+H)+; Anal. Calcd for
C27H27FN2O10: C, 58.06; H, 4.87; N, 5.02. Found: C, 58.10; H,
4.93; N, 5.05.
4.1.1. General procedure for the preparation of compounds 6a–
6j
4.1.1.2. 4-(40-Bromophenyl)-3-cyano-6-[(200,300,400,600-tetra-O-ace-
tyl-b-D-glucopyranosyl)methyl]pyridone (6b). Compound 6b
To a stirring solution of one of the selected (E)-1-(b-
pyranosyl)-4-(aryl)but-3-en-2-ones (5a–5j) (3.04 mmol) and
cyanoacetamide (3.04 mmol) in DMSO (10 mL), was added
D
-gluco-
was obtained by the reaction of compound 5b (0.69 g, 1.79 mmol),
cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g, 7.14 mmol),
pyridine (10 mL), and Ac2O (1.0 mL, 10.70 mmol) as a white solid

1196
S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
(0.6 g, yield 54%); mp >240 °C; Rf 0.5 (2:98, MeOH–CHCl3); ½a D25
ꢁ
white solid (0.58 g, yield 54%); mp >240 °C; Rf 0.5 (2:98, MeOH–
ꢀ118.36 (c 0.1, CHCl3); IR (KBr): mmax 3466 (NH stretching), 2225
(C„N stretching), 1753 (O–C@O stretching), 1660 cmꢀ1 (N–C@O
stretching); 1H NMR (300 MHz, DMSO-d6): d 12.5 (br s, 1H, NH),
7.61–7.56 (m, 2H, ArH), 7.48–7.44 (m, 2H, ArH), 6.20 (s, 1H, H-5),
5.17 (t, 1H, J = 9.3 Hz, CH), 5.01–4.87 (m, 2H, 2 ꢂ CH), 4.10–4.07
(m, 2H, H-100, H-600a), 3.84–3.83 (m, 1H, H-600b), 3.68–3.67 (m, 1H,
H-500), 2.84–2.75 (m, 2H, –CH2–), 2.01, 1.97, 1.94, 1.89 (each singlet,
12H, 4 ꢂ –OCOCH3); 13C NMR (50 MHz, DMSO-d6): d 170.4, 170.3,
170.3, 169.8 (OCOCH3), 162.0, 149.7, 132.5 (ArH), 130.9, 130.0,
129.3, 128.4 (ArH), 76.0, 75.9, 74.1, 71.7, 68.7 (OCH2), 35.9 (CH2),
21.0, 20.9 (CH3); ESIMS: m/z 621 (M+2)+; Anal. Calcd for
CHCl3); ½a 2D5
ꢁ
+6.15 (c 0.1, CHCl3); IR (KBr): mmax 3470 (NH stretch-
ing), 2225 (C„N stretching), 1753 (O–C@O stretching), 1660 cmꢀ1
(N–C@O stretching); 1H NMR (300 MHz, CDCl3): d 12.7 (s, 1H, NH),
7.31–7.24 (m, 2H, ArH), 7.00–6.97 (m, 1H, ArH), 6.38 (s, 1H, H-5),
5.29 (t, 1H, J = 9.36 Hz, CH), 5.11 (t, 1H, J = 9.8 Hz, CH), 5.03 (t,
1H, J = 9.6 Hz, CH), 4.19 (d, 2H, J = 3.4 Hz, H-600), 4.05 (m, 1H, H-
100), 3.97 (s, 3H, OCH3), 3.96 (s, 3H, OCH3), 3.83–3.77 (m, 1H, H-
500), 3.03–2.87 (m, 2H, CH2), 2.09, 2.03, 2.00, 1.97 (each singlet,
12H, 4 ꢂ COCH3); 13C NMR (50 MHz, CDCl3): d 170.6, 170.1,
170.0, 169.4 (OCOCH3), 163.7, 160.5, 151.4, 149.1, 148.6 (ArCH),
127.8, 121.6 (ArCH), 116.0 (CN), 111.2, 109.3, 75.7 (CH), 75.3
(CH), 73.9 (CH), 71.2 (CH), 68.2 (CH), 61.9 (OCH2), 56.1 (OCH3),
56.0 (OCH3), 35.8 (CH2), 20.8, 20.5 (CH3); DART-HRMS: m/z calcd
for C29H32N2O12 (M+H)+ 601.2033; found: 601.2043; ESIMS: m/z
601 (M+H)+; Anal. Calcd for C29H32N2O12: C, 58.00; H, 5.37; N,
4.66. Found: C, 58.05; H, 5.40; N, 4.66.
C27H27BrN2O10: C, 52.35; H, 4.39; N, 4.52. Found: C, 52.30; H,
4.41; N, 4.57.
4.1.1.3. 4-(40-Benzyloxyphenyl)-3-cyano-6-[(200,300,400,600-tetra-O-
acetyl-b-
D
-glucopyranosyl)methyl]pyridone
(6c). Compound
6c was obtained by the reaction of compound 5c (0.74 g,
1.79 mmol), cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g,
7.14 mmol), pyridine (10 mL), and Ac2O (1.0 mL, 10.7 mmol) as a
white solid (0.6 g, yield 52%); mp 215–217 °C; Rf 0.5 (2:98,
4.1.1.6. 4-(40-Chlorophenyl)-3-cyano-6-[(200,300,400,600-tetra-O-ace
tyl-b-D-glucopyranosyl)methyl]pyridone (6f). Compound 6f
was obtained by the reaction of compound 5f (0.61 g, 1.79 mmol),
cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g, 7.14 mmol),
pyridine (10 mL) and Ac2O (1.0 mL, 10.70 mmol) as a white solid
MeOH–CHCl3); ½a D25
ꢁ
+25.68 (c 0.1, CHCl3); IR (KBr): mmax 3472
(NH stretching), 2225 (C„N stretching), 1753 (O–C@O stretching),
1660 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz, CDCl3): d 12.3
(br s, 1H, NH), 7.62–7.60 (m, 5H, ArH), 7.44–7.33 (m, 3H, ArH),
7.09–7.06 (m, 1H, ArH), 6.24 (s, 1H, H-5), 5.23–5.13 (m, 3H,
OCH2, and CH), 4.97 (t, 1H, J = 9.8 Hz, CH), 4.87 (t, 1H, J = 9.2 Hz,
CH), 4.19–4.04 (m, 2H, H-100, H-600a), 3.85–3.82 (m, 2H, H-600b, H-
500), 2.84–2.58 (m, 2H, CH2), 2.05, 2.03, 2.02, 2.01 (each singlet
4 ꢂ COCH3); 13C NMR (50 MHz, CDCl3): d 170.8, 170.4, 170.1,
169.7 (OCOCH3), 163.1, 161.1, 160.2, 149.3, 136.6, 130.2 (ArCH),
129.0, 128.5 (ArCH), 127.8 (ArCH), 116.6 (CN), 115.5 (ArCH),
108.4, 99.4, 74.2, 73.0, 72.6, 71.7, 70.4, 68.7, 62.4 (OCH2), 35.9
(CH2), 21.0, 20.9, 20.8 (CH3); ESIMS: m/z 647 (M+H)+; Anal. Calcd
for C34H34N2O11: C, 63.15; H, 5.30; N, 4.33. Found: C, 63.19; H,
5.33; N, 4.38.
(0.6 g, yield 58%); mp >240 °C; Rf 0.5 (2:98, MeOH–CHCl3); ½a D25
ꢁ
ꢀ11.3 (c 0.1, CHCl3); IR (KBr): mmax 3472 (NH stretching), 2226
(C„N stretching), 1752 (O–C@O stretching), 1661 cmꢀ1 (N–C@O
stretching); 1H NMR (300 MHz, CDCl3): d 12.4 (s, 1H, NH), 7.54
(d, 2H, J = 8.5 Hz, ArH), 7.44 (d, 2H, J = 8.5 Hz, ArH), 6.19 (s, 1H,
H-5), 5.19 (t, 1H, J = 9.8 Hz, CH), 5.01 (t, 1H, J = 9.8 Hz, CH), 4.87
(t, 1H, J = 9.5 Hz, CH), 4.13–4.03 (m, 2H, H-600), 3.84–3.77 (m, 1H,
H-100), 3.69–3.64 (m, 1H, H-500), 2.88 (dd, 1H, J1 = 3.2 Hz,
J2 = 14.7 Hz, CHaH), 2.757 (dd, 1H, J1 = 8.2 Hz, J2 = 14.7 Hz, CHbH),
2.01, 1.97, 1.94, 1.88 (each singlet, 4xCOCH3); 13C NMR (50 MHz,
CDCl3): d 175.3, 175.0, 174.7, 174.4 (OCOCH3), 167.1, 164.0,
155.5, 141.6, 139.7, 134.7 (ArCH), 112.0 (ArCH), 121.1 (CN),
104.8, 80.5 (CH), 78.9 (CH), 78.9 (CH), 76.5 (CH), 73.5 (CH), 67.2
(OCH2), 40.5 (CH2), 25.8 (CH3), 25.7 (CH3), 25.65 (CH3); DART-
4.1.1.4.
O-acetyl-b-
3-Cyano-4-(40-methoxyphenyl)-6-[(200,300,400,600-tetra-
-glucopyranosyl)methyl]pyridone (6d). Compound
HRMS: m/z calcd for C
27H27ClN2O10 (M+) 575.1450; found:
D
575.1432, ESIMS: m/z 575 (M+H)+; Anal. Calcd for C27H27ClN2O10
:
6d was obtained by the reaction of compound 5d (0.60 g,
1.79 mmol), cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.60 g,
7.14 mmol), pyridine (10 mL), and Ac2O (1.0 mL, 10.70 mmol) as
a white solid (0.6 g, yield 60%); mp >240 °C; Rf 0.5 (2:98, MeOH–
C, 56.40; H, 4.73; N, 4.87. Found: C, 56.45; H, 4.77; N, 4.90.
4.1.1.7. 3-Cyano-4-phenyl-6-[(200,300,400,600-tetra-O-acetyl-b-
D-glu-
copyranosyl)methyl]pyridone (6g). Compound 6g was obtained
by the reaction of compound 5g (0.55 g, 1.79 mmol), cyanoacet-
amide (0.15 g, 1.79 mmol), t-BuOK (0.80 g, 7.14 mmol), pyridine
(10 mL) and Ac2O (1.0 mL, 10.70 mmol) as a white solid (0.52 g,
CHCl3); ½a 2D5
ꢁ
+16.4 (c 0.1, CHCl3); IR (KBr): mmax 3470 (NH stretch-
ing), 2224 (C„N stretching), 1754 (O–C@O stretching), 1658 cmꢀ1
(N–C@O stretching); 1H NMR (300 MHz, DMSO) d = 7.62 (d, 2H,
J = 8.7 Hz, ArH), 7.02 (d, 2H, J = 8.7 Hz, ArH), 6.26 (s, 1H, H-5),
5.24 (t, 1H, J = 9.36 Hz, CH), 4.97 (t, 1H, J = 9.8 Hz, CH), 4.87 (t,
1H, J = 9.5 Hz, CH), 4.18 (dd, 1H, J1 = 5.6 Hz, J2 = 12.3 Hz, H-600a),
4.04 (dd, 1H, J2 = 10.5 Hz, H-600b), 3.91—3.87 (m, 4H, H-100, OCH3),
3.78–3.73 (m, 1H, H-500), 2.88 (dd, 1H, J1 = 2.9 Hz, J2 = 14.7 Hz,
CHaH), 2.75 (dd, 1H, J1 = 8.5 Hz, J2 = 14.7 Hz, CHbH), 2.04, 2.01,
1.98, 1.86 (each singlet 4 ꢂ COCH3); 13C NMR (50 MHz, CDCl3): d
175.1, 174.8, 174.7, 174.5(OCOCH3), 167.2, 166.6, 164.7, 155.6,
135.1 (ArCH), 133.5 (ArCH), 122.0 (ArCH), 112.4 (CN), 101.2, 80.1
(CH), 78.8 (CH), 78.2 (CH), 77.3 (CH), 76.8 (CH), 67.5 (OCH2), 60.7
(OCH3), 40.4(OCH2), 25.9 (CH3), 25.8 (CH3), 25.6 (CH3); DART-
yield 54%); mp >240 °C; Rf 0.5 (2:98, MeOH–CHCl3); ½a D25
ꢀ39.1
ꢁ
(c 0.1, CHCl3); IR (KBr): mmax 3474 (NH stretching), 2227 (C„N
stretching), 1756 (O–C@O stretching), 1659 cmꢀ1 (N–C@O stretch-
ing); 1H NMR (300 MHz, DMSO-d6): d 12.5 (s, 1H, NH), 7.60–7.55
(m, 5H, ArH), 6.37 (s, 1H, H-5), 5.27 (t, 1H, J = 9.3 Hz, CH), 4.90–
4.73 (m, 2H, 2 ꢂ CH), 4.08–3.91 (m, 4H, H-600, H-100, H-500), 2.92
(m, 2H, CH2), 1.97, 1.92, 1.73 (12H, 4 ꢂ COCH3); 13C NMR
(50 MHz, CDCl3): d 170.7, 170.5, 170.3, 170.2 (OCOCH3), 162.1,
160.7, 152.3, 136.9, 131.3, 129.7, 128.8 (ArCH), 117.2 (CN), 107.7,
75.1 (CH), 74.0 (CH), 72.1 (CH), 69.2 (CH), 63.0 (CH), 63.0 (OCH2),
35.5 (CH2), 21.3, 21.2, 21.1, 20.9 (CH3); DART-HRMS: m/z calcd
for C27H29N2O10 (M+) 541.1822; found: 541.1829; ESIMS: m/z
541 (M+H)+; Anal. Calcd for C27H27ClN2O10: C, 60.00; H, 5.22; N,
5.18. Found: C, 60.04; H, 5.25; N, 5.15.
HRMS: m/z calcd for
C
28H30N2O11 (M+H)+ 571.1931; found:
571.1927; ESIMS: m/z 571 (M+H)+; Anal. Calcd for C28H30N2O11
:
C, 58.94; H, 5.30; N, 4.91. Found: C, 58.90; H, 5.37; N, 4.97.
4.1.1.5. 3-Cyano-4-(30,40-dimethoxyphenyl)-6-[(200,300,400,600tetra-
O-acetyl-b- -glucopyranosyl)methyl]pyridone (6e). Compound
6e was obtained by the reaction of compound 5e (0.65 g,
1.79 mmol), cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g,
7.14 mmol), pyridine (10 mL) and Ac2O (1.0 mL, 10.70 mmol) as a
4.1.1.8. 3-Cyano-6-[(200,300,400,600-tetra-O-acetyl-b-
D-glucopyrano-
D
syl)methyl]-4-(30,40,50-trimethoxyphenyl)-pyridone (6h). Com-
pound 6h was obtained by the reaction of compound 5h (0.82 g,
1.79 mmol), cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g,
7.14 mmol), pyridine (10 mL) and Ac2O (1.0 mL, 10.70 mmol) as a

S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
1197
white solid (0.7 g, yield 62%); mp >240 °C; Rf 0.5 (2:98, MeOH–
the residue so obtained was dissolved in MeOH and filtered
through a short column of silica gel to afford the products (7a–
7j). The compounds were crystallized with MeOH.
CHCl3); ½a 2D5
ꢁ
+68.97 (c 0.1, CHCl3); IR (KBr): mmax 3474 (NH stretch-
ing), 2223 (C„N stretching), 1753 (O–C@O stretching), 1658 cmꢀ1
(N–C@O stretching); 1H NMR (300 MHz, CDCl3): d 12.5 (s, 1H, NH),
6.91 (s, 2H, ArH), 6.36 (s, 1H, H-5), 5.29 (t, 1H, J = 9.36 Hz, CH), 5.11
(t, 1H, J = 9.8 Hz, CH), 5.02 (t, 1H, J = 9.6 Hz, CH), 4.23 (m, 2H, H-600),
4.00 (m, 1H, H-100), 3.98 (s, 3H, OCH3), 3.95 (s, 3H, OCH3), 3.93 (s,
3H, OCH3), 3.81–3.78 (m, 1H, H-500), 3.03–2.86 (m, 2H, CH2), 2.18,
2.15, 2.10, 2.01 (each singlet, 12H, 4 ꢂ COCH3); 13C NMR
(50 MHz, CDCl3): d 171.0, 170.6, 170.4, 169.9 (OCOCH3), 163.9,
161.2, 153.8, 149.2, 140.6, 131.0, 116.2 (CN), 109.8, 106.1, 76.1
(CH), 75.6 (CH), 74.3 (CH), 71.4 (CH), 68.5 (CH), 62.2 (OCH2), 61.4
(OCH3), 56.8 (OCH3), 56.0 (OCH3), 36.1 (CH2), 21.2, 21.0, 20.9
(CH3); DART-HRMS: m/z calcd for C30H34N2O13 (M+) 631.2139;
found: 631.2146; ESIMS: m/z 631 (M+H)+; Anal. Calcd for
4.1.2.1.
3-Cyano-4-(40-fluorophenyl)-6-[(b-
D-glucopyrano-
syl)methyl]pyridone (7a). Compound 7a was obtained by reac-
tion of 3-cyano-4-(40-fluorophenyl)-6-[(200,300,400,600-tetra-O-acetyl-
b-D-glucopyranosyl)methyl]pyridone (6a) (0.4 g, 7.16 mmol) and
MeOH/NaOMe (10 mL, 10 mol %). White solid (0.26 g, yield 71%);
mp >240 °C; Rf 0.5 (2:8, MeOH–CHCl3); ½a D25
ꢀ49.4 (c 0.1, MeOH);
ꢁ
IR (KBr): mmax 3467–3412 (OH and NH stretching), 2223 (C„N
stretching), 1690 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz,
DMSO-d6): d 12.3 (br s, 1H, NH), 7.74–7.70 (m, 2H, ArH), 7.39–
7.33 (m, 2H, ArH), 6.44 (s, 1H, H-5), 4.91–4.60 (m, 4H, 4 ꢂ OH),
3.47–3.36 (m, 2H), 3.23–3.17 (m, 2H), 3.11–3.01 (m, 3H), 2.97–
2.91 (m, 1H, –CHaH–), 2.70 (dd, 1H, J1 = 8.22 Hz, J2 = 15.2 Hz, –
CHbH–); 13C NMR (50 MHz, DMSO-d6): d 166.5, 162.1, 159.5,
154.1, 133.4, 133.3, 131.2 (ArCH), 131.5 (ArCH), 117.5, 116.8,
116.4 (CN), 107.7, 81.4, 78.7, 77.8, 74.1, 71.1, 62.1 (OCH2), 36.1
(CH2); DART-HRMS: m/z calcd for C19H20FN2O6 (M+) 391.1305;
found: 391.1310; ESIMS: m/z 391 (M+H)+; Anal. Calcd for
C30H34N2O13: C, 57.14; H, 5.43; N, 4.44. Found: C, 57.11; H, 5.38;
N, 4.40.
4.1.1.9. 3-Cyano-4-(20-napthyl)-6-[(200,300,400,600-tetra-O-acetyl-b-
D
-glucopyranosyl)methyl]pyridone (6i). Compound 6i was
obtained by the reaction of compound 5i (0.63 g, 1.79 mmol), cya-
noacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g, 7.14 mmol), pyr-
idine (10 mL) and Ac2O (1.0 mL, 10.7 mmol) as a white solid
C19H19FN2O6: C, 58.46; H, 4.91; N, 7.18. Found: C, 58.43; H, 4.94;
N, 7.14.
(0.61 g, yield 58%); mp >240 °C; Rf 0.5 (2:98, MeOH–CHCl3); ½a D25
ꢁ
ꢀ151 (c 0.1, MeOH); IR (KBr): mmax 3461 (NH stretching), 2228
(C„N stretching), 1748 (O–C@O stretching), 1663 cmꢀ1 (N–C@O
stretching); 1H NMR (300 MHz, CDCl3): d 8.01–7.93 (m, 2H, ArH),
7.72–7.63 (m, 1H, ArH), 7.58–7.50 (m, 4H, ArH), 6.36 (s, 1H, H-5),
5.24 (t, 1H, J = 8.9 Hz, CH), 4.93 (t, 1H, J = 8.5 Hz, CH), 4.84 (t, 1H,
J = 9.2 Hz, CH), 4.03–3.96 (m, 3H), 3.91–3.81 (m, 1H), 2.90–2.89
(m, 1H), 2.72–2.63 (m, 1H), 2.12, 2.04, 2.01, 1.96 (each singlet,
12H, 4 ꢂ COCH3); DART-HRMS: m/z calcd for C31H31N2O10 (M+)
591.1978; found: 591.1971; ESIMS: m/z 591 (M+H)+; Anal. Calcd
for C31H30N2O10: C, 63.05; H, 5.12; N, 4.74. Found: C, 63.00; H,
5.15; N, 4.80.
4.1.2.2. 4-(40-Bromophenyl)-3-cyano-6-[(b-
D-glucopyranosyl)-
methyl]pyridone (7b). Compound 7b was obtained by the reac-
tion of compound 6b (0.50 g, 0.81 mmol), NaOMe/MeOH (10 mL,
10 mol %) as a white solid (0.28 g, yield 77%); mp 225–227 °C; Rf
0.5 (2:8, MeOH–CHCl3); ½a D25
ꢀ117 (c 0.1, MeOH); IR (KBr): mmax
ꢁ
3503–3353 (OH and NH stretching), 2230 (C„N stretching),
1682 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz, D2O): d 7.72
(d, 1H, J = 8.5 Hz, ArH), 7.63–7.51 (m, 3H, ArH), 6.42 (s, 1H, H-5),
3.62–3.59 (m, 2H), 3.20–3.01 (m, 4H), 2.95–2.92 (m, 1H), 2.72–
2.70 (m, 1H, CHbH); DART-HRMS: m/z calcd for C19H20BrN2O6
(M+) 451.0504; found: 451.0508; ESIMS: m/z 451 (M+H)+; Anal.
Calcd for C19H19BrN2O6: C, 50.57; H, 4.24; N, 6.21. Found: C,
50.52; H, 4.20; N, 6.15.
4.1.1.10. 4-(40-O-Acetylphenyl)-3-cyano-6-[(200,300,400,600-tetra-O-
acetyl-b-D-glucopyranosyl)methyl]pyridone (6j). Compound 6j
was obtained by the reaction of compound 5j (0.58 g, 1.79 mmol),
cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g, 7.14 mmol),
pyridine (10 mL) and Ac2O (1.0 mL, 10.7 mmol) as a white solid
4.1.2.3.
4-(40-Benzyloxyphenyl)-3-cyano-6-(b-
D-glucopyrano-
syl)methyl]pyridone (7c). Compound 7c was obtained by the
reaction of compound 6c (0.40 g, 0.62 mmol), NaOMe/MeOH
(10 mL, 10 mol %) as a white solid (0.2 g, yield 67%); mp >218–
(0.6 g, yield 56%); mp >240 °C; Rf 0.5 (2:98, MeOH–CHCl3); ½a D25
ꢁ
+18.5 (c 0.1, CHCl3); IR (KBr): mmax 3428 (NH stretching), 2215
(C„N stretching), 1632 cmꢀ1 (N–C@O stretching); 1H NMR
(300 MHz, CDCl3): d 12.3 (s, 1H, NH), 7.70–7.68 (d, 2H, J = 8.3 Hz,
ArH), 6.28–7-26 (d, 2H, J = 8.3 Hz, ArH), 6.35 (s, 1H, H-5), 5.28 (t,
1H, J = 7.2 Hz, CH), 5.12–4.94 (m, 2H, 2xCH), 4.19 (m, 2H, H-600),
3.95 (m, 1H, H-100), 3.80 (m, 1H, H-500), 2.94–2.84 (m, 2H, CH2),
2.37, 2.11, 2.06, 2.04, 2.01 (15H, 5 ꢂ COCH3); 13C NMR (50 MHz,
CDCl3): d 161.8, 136.5, 130.8, 129.5, 122.3, 116.3 (CN), 109.1,
75.8 (CH), 75.4 (CH), 73.8 (CH), 71.1 (CH), 68.2 (CH), 61.9 (OCH2),
35.6 (CH2), 21.1, 20.7, 20.5 (CH3); DART-HRMS: m/z calcd for
220 °C; Rf 0.5 (2:8, MeOH–CHCl3); ½a D25
ꢀ45.9 (c 0.1, MeOH); IR
ꢁ
(KBr): mmax 3346–3279 (OH and NH stretching), 2224 (C„N
stretching), 1690 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz,
DMSO-d6): d 12.2 (br s, 1H, NH), 7.67–7.64 (d, 2H, ArH), 7.49–
7.34 (m, 5H, ArH), 7.17 (d, 2H, J = 8.8 Hz), 6.44 (s, 1H, H-5), 5.27
(d, 1H, J = 5.3 Hz, OH), 5.19 (s, 2H, OCH2), 5.05 (d, 1H, J = 4.3 Hz,
OH), 5.00 (d, 1H, J = 4.9 Hz, OH), 4.52 (t, 1H, J = 5.7 Hz, OH), 3.65
(dd, J1 = 6.2 Hz, J2 = 10.5 Hz, 1H, H6a00), 3.45–3.39 (m, 1H), 3.17–
3.01 (m, 6H), 2.95 (dd, 1H, J1 = 5.37 Hz, J2 = 8.91 Hz, –CHbH–); 13C
NMR (50 MHz, DMSO-d6): d 162.4, 160.9, 160.0, 153.5, 137.5,
130.8 (ArCH), 129.3 (ArCH), 129.1, 128.8 (ArCH), 128.6 (ArCH),
117.8, 115.8 (CN), 107.5, 81.3 (CH), 78.7 (CH), 77.8 (CH), 74.0
(CH), 71.1 (CH), 70.3 (OCH2), 62.1 (OCH2), 36.0 (CH2); DART-HRMS:
m/z calcd for C26H27N2O7 (M+) 479.1818; found: 479.1830; ESIMS:
m/z 479 (M+H)+; Anal. Calcd for C26H26N2O7: C, 65.26; H, 5.48; N,
5.85. Found: C, 65.22; H, 5.50; N, 5.80.
C
29H31N2O12 (M+H)+ 599.1877; found: 599.1895; ESIMS: m/z 599
(M+H)+. Anal. Calcd for C29H30N2O12: C, 58.19; H, 5.05; N, 4.68.
Found: C, 58.23; H, 5.00; N, 4.60.
4.1.2. General procedure for synthesis of compounds 7a–7j
To a stirring solution of 3-cyano-4-(aryl)-6-[(200,300,400,600-tetra-
O-acetyl-b-
D
-glucopyranosyl)methyl]pyridones
(6a–6j)
(7.16
mmol) in MeOH at 25 °C, was added a solution of NaOMe (10 mL,
10 mol %). The reaction mixture was stirred until the disappear-
ance of the starting material was confirmed (TLC). After completion
of the reaction, the reaction mixture was neutralized by 2% aq HCl
solution at 0 °C and filtered, and the filtrate was concentrated in
vacuum to give a residual mass that was triturated with a mixture
of CHCl3 and MeOH and filtered. The filtrate was evaporated, and
4.1.2.4.
3-Cyano-4-(40-methoxyphenyl)-6-[(b-
D-glucopyrano-
syl)methyl]pyridone (7d). Compound 7d was obtained by the
reaction of compound 6d (0.40 g, 0.70 mmol), MeOH/NaOMe
(10 mL, 10 mol %) as a white solid (0.2 g, yield 71%); mp >240 °C;
Rf 0.5 (2:8, MeOH–CHCl3); ½a D25
ꢀ142 (c 0.1, MeOH); IR (KBr): mmax
ꢁ
3380–3272 (OH and NH stretching), 2219 (C„N stretching),
1653 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz, DMSO-d6): d

1198
S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
12.2 (br s, 1H, NH), 7.67 (d, 2H, J = 8.8 Hz, ArH), 7.10 (d, 2H,
J = 8.8 Hz, ArH), 5.30 (d, 1H, J = 5.2 Hz, OH), 5.10 (d, 1H, J = 4.4 Hz,
OH), 5.04 (d, 1H, J = 4.8 Hz, OH), 4.54 (t, 1H, J = 5.8 Hz, OH), 3.83
(s, 3H, OCH3), 3.60 (dd, 1H, J = 6.3 Hz, H-6a00), 3.43–3.38 (m, 2H),
3.18–2.91(m, 5H), 2.70 (dd, 1H, J1 = 8.22 Hz, –CHbH–); 13C NMR
(50 MHz, DMSO-d6): d 167.1, 166.6, 164.8, 158.2, 135.5, 133.6
(ArCH), 122.6, 119.8 (ArCH), 112.2 (CN), 86.1 (CH), 83.5 (CH),
82.6 (CH), 78.8 (CH), 75.9 (CH), 66.8 (OCH2), 61.0 (OCH3), 40.8
(CH2); DART-HRMS: m/z calcd for C20H22N2O7 (M+) 403.1505;
found: 403.1515; ESIMS: m/z 403 (M+H)+; Anal. Calcd for
4.1.2.8. 3-Cyano-6-[(b-D
-glucopyranosyl)methyl]-4-(30,40,50-tri-
methoxyphenyl)-pyridone (7h). Compound 7h was obtained
by the reaction of compound 6h (0.40 g, 0.63 mmol), NaOMe/
MeOH (10 mL, 10 mol %) as a white solid (0.2 g, yield 68%); mp
>240 °C; Rf 0.5 (2.5:7.5, MeOH–CHCl3); ½a D25
ꢀ138 (c 0.1c, MeOH);
ꢁ
IR (KBr): mmax 3633–3356 (OH and NH stretching), 2225 (C„N
stretching), 1621 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz,
DMSO): d 12.2 (br s, 1H, NH), 6.96 (s, 2H, ArH), 6.51 (s, 1H, H-5),
5.28 (d, 1H, J = 5.1 Hz, OH), 5.05 (d, 1H, J = 3.4 Hz, OH), 4.99 (d,
1H, J = 5.6 Hz, OH), 4.49 (t, 1H, J = 5.6 Hz, OH), 3.84 (m, 6H,
2 ꢂ OCH3), 3.73 (s, 3H, OCH3), 3.64–3.59 (m, 2H, H-100, H-6a00),
3.47–3.35 (m, 2H, J1), 3.17–2.99 (m, 4H), 2.96 (m, 1H, –CHbH–);
13C NMR (50 MHz, DMSO) d 162.2, 160.5, 153.6, 132.0, 117.8
(CN), 106.7, 81.3 (CH), 78.7 (CH), 77.7 (CH), 74.1 (CH), 71.0 (CH),
62.1 (OCH2), 61.02 (OCH3), 56.9 (OCH3), 39.9 (CH2); DART-HRMS:
m/z calcd for C22H27N2O9 (M+H)+ 463.1716; found: 463.1713;
ESIMS: m/z 463 (M+H)+; Anal. Calcd for C22H26N2O9: C, 57.14; H,
5.67; N, 6.06. Found: C, 57.10; H, 5.64; N, 6.00.
C20H22N2O7: C, 59.70; H, 5.51; N, 6.96. Found: C, 59.73; H, 5.48;
N, 6.90.
4.1.2.5. 3-Cyano-4-(30,40-dimethoxyphenyl)-6-[(b-
D-glucopyran-
osyl)methyl]pyridone (7e). Compound 7e was obtained by the
reaction of compound 6e (0.40 g, 0.67 mmol), NaOMe/MeOH
(10 mL, 10 mol %) as a white solid (0.2 g, yield 64%); mp >240 °C;
Rf 0.5 (2.5:7.5, MeOH–CHCl3); ½a D25
ꢀ124 (c 0.1c, MeOH); IR
ꢁ
(KBr): mmax 3631–3356 (OH and NH stretching), 2229 (C„N
stretching), 1619 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz,
DMSO-d6): d 12.1 (br s, 1H, NH), 7.29–7.24 (m, 2H, ArH), 7.11–
7.08 (m, 2H, ArH), 6.48 (s, 1H, H-5), 5.23 (d, 1H, J = 5.3 Hz, OH),
4.99 (d, 1H, J = 4.2 Hz, OH), 4.95 (d, 1H, J = 4.9 Hz, OH), 4.49 (t,
1H, J = 5.6 Hz, OH), 4.12–4.11 (m, 1H, H-100), 3.83 (s, 3H, OCH3),
3.82 (s, 3H, OCH3), 3.60 (dd, 1H, J1 = 5.6 Hz, J2 = 11.0 Hz, H-6a00),
3.46 (dd, 1H, J1 = 8.7 Hz, H-6b00), 3.17–2.91 (m, 5H), 2.70 (dd, 1H,
J1 = 8.28 Hz, J2 = 15.21 Hz, –CHbH–); 13C NMR (50 MHz, DMSO-
d6): d 162.4, 160.3, 153.3, 151.5, 149.4, 129.0, 122.2, (ArCH),
122.6, 112.5, 112.4 (ArCH), 117.9 (CN), 81.3(CH), 78.7 (CH), 77.8
(CH), 74.1 (CH), 71.2 (CH), 62.1 (OCH2), 56.5 (OCH3), 36.0 (CH2);
DART-HRMS: m/z calcd for C21H25N2O8 (M+H)+ 433.1610; found:
433.1618; ESIMS: m/z 433 (M+H)+; Anal. Calcd for C21H24N2O8: C,
58.33; H, 5.59; N, 6.48. Found: C, 58.30; H, 5.53; N, 6.44.
4.1.2.9. 3-Cyano-4-(20-napthyl)-6-[(b-
D-glucopyranosyl)methyl]-
pyridone (7i). Compound 7i was obtained by the reaction of
compound 6i (0.40 g, 0.67 mmol), NaOMe/MeOH (10 mL,
10 mol %) as a white solid (0.2 g, yield 70%); mp >240 °C; Rf 0.5
(2.5:7.5, MeOH–CHCl3); ½a D25
ꢀ156 (c 0.1, MeOH); IR (KBr): mmax
ꢁ
3633–3319 (OH and NH stretching), 2230 (C„N stretching),
1621 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz, DMSO-d6): d
8.20–8.03 (m, 2H, ArH), 7.74–7.71 (m, 1H, ArH), 7.65–7.52 (m,
4H, ArH), 6.39 (s, 1H, H-5), 5.11–4.47 (m, 4H, 4 ꢂ OH), 3.61–3.57
(m, 2H), 3.39 (m, 1H), 3.15–3.05 (m, 4H), 2.96–2.90 (m, 2H), 2.76
(m, 1H, –CHbH–); 13C NMR (50 MHz, DMSO-d6): d 162.4, 160.4,
154.4, 135.0, 133.9, 130.5, 129.3, 128.0, 127.3, 126.2, 125.6, 117.8
(CN), 109.2, 81.3 (CH), 78.7 (CH), 78.0 (CH), 77.7 (CH), 74.4 (CH),
62.0 (OCH2), 36.3 (CH2); DART-HRMS: m/z calcd for C23H22N2O6
(M+H)+ 423.1556; found: 423.1561; ESIMS: m/z 423 (M+H)+; Anal.
Calcd for C23H22N2O6: C, 65.39; H, 5.25; N, 6.63. Found: C, 65.35; H,
5.28; N, 6.60.
4.1.2.6. 4-(40-Chlorophenyl)-3-cyano-6-[(b-
D-glucopyranosyl)-
methyl]pyridone (7f). Compound 7f was obtained by the reac-
tion of compound 6f (0.40 g, 1.78 mmol), NaOMe/MeOH (10 mL,
10 mol %) as a white solid (0.2 g, yield 71%); mp >240 °C; Rf 0.5
4.1.2.10.
3-Cyano-4-(40-hydroxyphenyl)-6-[(b-
D-glucopyrano-
(2.5:7.5, MeOH–CHCl3); ½a D25
ꢁ
ꢀ59.3 (c 0.1, MeOH); IR (KBr): mmax
syl)methyl]pyridone (7j). Compound 7j was obtained by the
reaction of compound 6j (0.40 g, 0.67 mmol), NaOMe/MeOH
3667–3374 (OH and NH stretching), 2223 (C„N stretching),
1649 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz, D2O):
d = 7.65–7.54 (m, 4H, ArH), 6.42 (s, 1H, H-5), 3.62–3.56 (m, 1H),
3.39–3.20 (m, 2H), 3.16–2.86 (m, 5H), 2.73 (m, 1H, J1 = 8.4 Hz,
J2 = 15.3 Hz, –CHbH–); 13C NMR (50 MHz, DMSO-d6): d 162.1,
159.3, 154.3, 136.0, 135.7, 130.9, 129.7, 117.3 (CN), 107.5, 81.4
(CH), 78.7 (CH), 77.8 (CH), 74.1 (CH), 71.1 (CH), 62.0 (OCH2), 36.1
(CH2); DART-HRMS: m/z calcd for C19H19ClN2O6 (M+H)+
407.1009; found: 407.1020; ESIMS: m/z 407 (M+H)+; Anal. Calcd
for C19H19ClN2O6: C, 56.09; H, 4.71; N, 6.89. Found: C, 56.00; H,
4.65; N, 6.83.
(10 mL, 10 mol %) as
a white solid (0.20 g, yield 77%); mp
>240 °C; Rf 0.5 (2.5:7.5, MeOH–CHCl3); ½a D25
ꢀ134 (c 0.1, MeOH);
ꢁ
IR (KBr): mmax 3667–3363 (OH and NH stretching), 2225 (C„N
stretching), 1648 cmꢀ1 (N–C@O stretching); 1H NMR (300 MHz,
DMSO-d6): d 7.55 (d, 2H, J = 8.6 Hz, ArH), 6.94 (d, 2H, J = 8.6 Hz,
ArH), 6.40 (s, 1H, H-5), 5.06–5.00 (m, 3H, 3 ꢂ OH), 4.49 (br s, 1H,
OH), 3.63–3.59 (m, 1H), 3.20–3.02 (m, 8H), 2.95–2.89 (m, 1H),
2.69–2.61 (dd, 1H, J1 = 8.1 Hz, J2 = 15.2 Hz, –CHbH–); DART-HRMS:
m/z calcd for C19H20N2O7 (M+H)+ 389.1348; found: 389.1335;
ESIMS: m/z 389 (M+H)+; Anal. Calcd for C19H20N2O7: C, 58.76; H,
5.19; N, 7.21. Found: C, 58.73; H, 5.15; N, 7.18.
4.1.2.7. 3-Cyano-4-phenyl-6-[(b-D-glucopyranosyl)methyl]pyri-
done (7g). Compound 7g was obtained by the reaction of com-
pound 6g (0.40 g, 0.74 mmol), NaOMe/MeOH (10 mL, 10 mol %) as
a white solid (0.21 g, yield 76%); mp >240 °C; Rf 0.5 (2.5:7.5,
4.1.2.11. 3-Cyano-6-[(200,300,600,2000,3000,4000,6000-hepta-O-acetyl-b-cel-
lobiosyl)methyl]-4-(20-napthyl)-pyridone (9). Compound
9
was obtained by the reaction of compound 8 (0.87 g, 1.79 mmol),
cyanoacetamide (0.15 g, 1.79 mmol), t-BuOK (0.80 g, 7.14 mmol),
pyridine (10 mL), and Ac2O (1.0 mL, 10.70 mmol) as a white solid
(0.6 g, yield 54%); mp 155–157 °C; Rf 0.5 (4:96, MeOH–CHCl3);
MeOH–CHCl3); ½a D25
ꢀ141 (c 0.1, MeOH); IR (KBr): mmax 3667–
ꢁ
3268 (OH and NH stretching), 2225 (C„N stretching), 1617 cmꢀ1
(N–C@O stretching); 1H NMR (200 MHz, DMSO-d6): d 12.3 (br s,
1H, NH), 7.63–7.53 (m, 5H, ArH), 6.43 (s, 1H, H-5), 3.62–3.4 (m,
4H, 4 ꢂ OH), 3.20–3.04 (m, 6H), 2.95 (m, 2H), 2.73 (m, 1H,
J1 = 7.8 Hz, J2 = 14.9 Hz, –CHbH–); 13C NMR (50 MHz, DMSO-d6): d
161.8, 136.5, 130.8, 129.2, 128.5, 116.3 (CN), 107.2, 80.9 (CH), 78.3
(CH), 77.4 (CH), 73.6 (CH), 73.6 (CH), 621.6 (OCH2), 36.8 (CH2);
DART-HRMS: m/z calcd for C19H21N2O6 (M+H)+ 373.1399; found:
373.1394; ESIMS: m/z 373 (M+H)+; Anal. Calcd for C19H20N2O6: C,
61.28; H, 5.41; N, 7.52. Found: C, 61.25; H, 5.37; N, 7.49.
½
a 2D5
ꢁ
ꢀ138.12 (c 0.1, CHCl3); IR (KBr): mmax 3414 (NH stretching),
2245 (C„N stretching), 1747 (O–C@O stretching), 1644 cmꢀ1 (N–
C@O stretching); 1H NMR (300 MHz, DMSO-d6): d 12.7 (s, 1H,
NH), 8.18 (s, 1H, ArH), 7.94–7.89 (m, 4H, ArH), 7.73–7.56 (m, 2H,
ArH), 6.43 (s, 1H, H-5), 5.20–5.04 (5H), 4.61–4.52 (2H), 4.38–4.35
(1H), 4.17–3.87 (3H), 3.78–3.66 (3H), 3.01–2.89 (m, 2H), 2.08–
1.78(21H, 7 ꢂ COCH3); 13C NMR (50 MHz, CDCl3): d 170.0, 169.8,
169.5, 168.9, 168.7 (OCOCH3), 163.4, 160.7, 134.0, 132.8, 128.8,

S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
1199
127.7, 126.9, 124.7, 109.4 (CN), 100.6 (C-1000), 76.3, 75.2, 73.6, 72.9,
71.9, 71.5, 67.7 (CH), 61.7 (OCH2), 35.8 (CH2), 20.7, 20.5, 20.4;
ESIMS: m/z 879 (M+H)+; Anal. Calcd for C43H46N2O18: C, 58.77; H,
5.28; N, 3.19. Found: C, 58.70; H, 5.25; N, 3.24.
169.5 (OCOCH3), 162.0, 154.9, 135.4, 132.9 (ArCH), 131.0, 130.4
(ArCH), 129.6 (ArCH), 128.8(ArCH), 125.9, 123.7, 114.7 (CN), 76.3,
74.4, 72.0, 68.8, 62.3 (OCH2), 40.7 (CH2), 21.0, 20.9, 20.8, 20.7;
DART-HRMS: m/z calcd for C27H27BrClN2O9 (M+H)+ 637.0588;
found: 637.0608; ESIMS: m/z 638 (M+H)+; Anal. Calcd for
4.1.2.12. 6-[(b-Cellobiosyl)methyl]-3-cyano-4-(20-napthyl)pyri-
done (10). Compound 10 was obtained by the reaction of com-
pound 9 (0.50 g, 0.56 mmol), NaOMe/MeOH (10 mL, 10 mol %) as
a white solid (0.2 g, yield 60%); mp >240 °C; Rf 0.5 (3:7, MeOH–
C27H26BrClN2O9: C, 50.84; H, 4.11; N, 4.39. Found: C, 50.80; H,
4.09; N, 4.33.
4.1.3.3. 4-(40-Benzyloxyphenyl)-2-chloro-3-cyano-6-[(200,300,400,600-
tetra-O-acetyl-b-D-glucopyranosyl)methyl]pyridine (11c). Com-
pound 11c was obtained by the reaction of compound 6c
(0.50 g, 0.77 mmol), POCl3 (10 mL), and toluene (10 mL) as a
white solid (0.4 g, yield 77%); mp >240 °C; Rf 0.5 (2:8, MeOH–
CHCl3); ½a 2D5
ꢁ
+8.03 (c 0.1, MeOH); IR (KBr): mmax 3667–3380 (OH
and NH stretching), 2225 (C„N stretching), 1649 cmꢀ1 (N–C@O
stretching); 1H NMR (300 MHz, DMSO-d6): d 12.3 (br s, 1H, NH),
8.36 (s, 1H, ArH), 7.77–7.61 (m, 2H, ArH), 7.63–7.53 (m, 5H, ArH),
6.58 (s, 1H, H-5), 5.50–5.48 (m, 1H, OH), 5.29–5.27 (m, 1H, OH),
5.14–5.10 (m, 2H, OH), 4.80 (s, 1H, H-1000), 4.68–4.63 (m, 3H, OH),
4.27–4.24 (1H, m, H-100), 3.70–3.66 (m, 3H), 3.56–3.54 (m, 3H),
3.29–3.18 (m, 4H), 3.09–3.00 (m, 3H), 2.98 (m, 1H); ESIMS: m/z
585 (M+H)+; Anal. Calcd for C29H32N2O11: C, 59.58; H, 5.52; N,
4.79. Found: C, 59.55; H, 5.48; N, 4.70.
CHCl3); ½a 2D5
ꢁ
ꢀ48.4 (c 0.1, CHCl3); IR (KBr): mmax 2231 cmꢀ1
(C„N stretching); 1H NMR (300 MHz, CDCl3): d 7.63 (d, 2H,
J = 8.7 Hz, ArH), 7.44–7.36 (m, 5H, ArH), 7.29 (s, 1H, ArH),
7.15–7.12 (d, J = 8.7 Hz, 2H, ArH), 5.15 (s, 2H, OCH2), 5.22 (t,
1H, J = 9.36 Hz, CH), 5.09 (t, 1H, J = 9.66 Hz, CH), 4.94 (t, 1H,
J = 9.57 Hz, CH), 4.21 (dd, 1H, J = 7.11 Hz, J = 12.2 Hz, H-600a),
4.06–4.02 (m, 2H, H-100, H-600b), 3.66–3.62 (m, 1H, H-500), 3.05–
3.00 (m, 2H, CH2), 2.06, 2.05, 2.03, 1.90 (each singlet, 12H,
4 ꢂ OCOCH3); 13C NMR (50 MHz, CDCl3): d 170.4, 170.2, 169.8,
169.5 (OCOCH3), 161.4, 155.7, 154.1, 136.6, 130.5 (ArCH),
129.0 (ArCH), 128.6 (ArCH), 127.8 (ArCH), 127.7, 123.5 (ArCH),
115.9 (ArCH), 115.3 (CN), 107.0, 76.7, 74.4, 72.1, 70.5 (OCH2),
68.8, 62.2 (OCH2), 40.8 (CH2), 23.1, 21.0, 20.9, 20.8; DART-
4.1.3. General procedure for synthesis of compounds 11a–11e
To a stirring solution of 1:1 POCl3–toluene, was added 3-cyano-
4-(aryl)-6-[(200,300,400,600-tetra-O-acetyl-b-
D-glucopyranosyl)methyl]-
pyridone (6a–6e) (0.896 mmol) at room temperature. The reaction
mixture was then heated to 90–100 °C for 3 h with continuous
stirring. After completion of the reaction, the mixture was cooled,
poured into ice containing K2CO3, and the whole mixture was
neutralized (pH 7.0) with aq ammonia. The reaction mixture was
extracted with EtOAc, and the organic layer was evaporated under
reduced pressure to give 2-chloro-3-cyano-4-(aryl)-6-[(200,300,400,600-
HRMS: m/z calcd for
665.1897; ESIMS: m/z 665 (M+H)+; Anal. Calcd for
34H33ClN2O10: C, 61.40; H, 5.00; N, 4.21. Found: C, 61.38; H,
C
34H34ClN2O10 (M+) 665.1902; found:
C
5.09; N, 4.26.
tetra-O-acetyl-b-D-glucopyranosyl)methyl]pyridines (11a–11e) as
white solids.
4.1.3.4. 2-Chloro-3-cyano-4-(40-methoxyphenyl)-6-[(200,300,400,600-
tetra-O-acetyl-b-D-glucopyranosyl)methyl]pyridine (11d). Com-
pound 11d was obtained by the reaction of compound 6d (0.40 g,
4.1.3.1.
tetra-O-acetyl-b-
2-Chloro-3-cyano-4-(40-fluorophenyl)-6-[(200,300,400,600-
-glucopyranosyl)methyl]pyridine (11a). Com-
D
0.70 mmol), POCl3 (10 mL), and toluene (10 mL) as a white solid
pound 11a was obtained by the reaction of compound 6a (0.50 g,
0.896 mmol), POCl3 (10 mL), and toluene (10 mL), as a white solid
(0.3 g, yield 58%); mp 119–120 °C; Rf 0.5 (3:97, MeOH–CHCl3);
(0.3 g, yield 73%); mp >240 °C; Rf 0.5 (2:98, MeOH–CHCl3); ½a D25
ꢁ
ꢀ24.0 (c 0.1, MeOH); IR (KBr): mmax 2228 (C„N stretching),
1749 cmꢀ1 (O–C@O stretching); 1H NMR (300 MHz, CDCl3): d
7.61 (d, 1H, J = 8.3 Hz, ArH), 7.25 (s, 1H, ArH), 7.05 (d, 1H,
J = 8.3 Hz, ArH), 5.19 (t, 1H, J = 9.12 Hz, CH), 5.03 (t, 1H, J = 7.5 Hz,
CH), 4.90 (t, 1H, J = 9.63 Hz, CH), 4.19 (m, 1H, H-6a00), 4.03–3.97
(m, 2H, H-100, H-6b00), 3.88 (s, 3H, OCH3), 3.63–3.59 (m, 1H, H-500),
3.01–2.97 (m, 2H, CH2), 2.02–1.53 (12H, 4xOCOCH3); 13C NMR
(50 MHz, CDCl3): d 169.9, 169.8, 169.4, 169.1 (OCOCH3), 161.6,
155.4, 153.7, 130.0 (ArCH), 127.0, 123.1 (ArCH), 114.9 (ArCH),
114.6 (CN), 106.6, 76.4 (CH), 75.9 (CH), 74.0 (CH), 71.7 (CH), 68.4
(CH), 61.8 (OCH2), 55.3 (OCH3), 40.3 (CH2), 20.6, 20.5, 20.4, 20.3;
½
a 2D5
ꢁ
ꢀ120 (c 0.1, CHCl3); IR (KBr): mmax 2232 (C„N stretching),
1754 cmꢀ1 (O–C@O stretching); 1H NMR (300 MHz, CDCl3): d
7.66–7.61 (m, 2H, ArH), 7.28–7.22 (m, 3H, ArH), 5.20 (t, 1H,
J = 9.33 Hz, CH), 5.04 (t, 1H, J = 9.81 Hz, CH), 4.95 (t, 1H,
J = 9.63 Hz, CH), 4.13 (dd, 1H, J = 4.98 Hz, H-600a), 4.06–4.00 (m,
2H, H-100, H-600b), 3.52–3.64 (m, 1H, H-500), 3.05–2.99 (m, 2H, –
CH2–), 2.05, 2.04, 2.03, 1.91 (each singlet, 12H, 4 ꢂ OCOCH3); 13C
NMR (50 MHz, CDCl3): d 170.3, 170.2, 169.8, 169.5 (OCOCH3),
161.9, 155.0, 154.1, 131.3, 131.1 (ArCH), 130.9 (ArCH), 123.8,
117.1 (ArCH), 116.6 (ArCH), 114.8 (CN), 76.7, 76.3, 74.4, 72.1,
68.8, 62.1 (OCH2), 40.8 (CH2), 21.0, 20.9, 20.8 (CH3); DART-HRMS:
m/z calcd for C27H27ClFN2O9 (M+) 577.1389; found: 577.1395;
ESIMS: m/z 577 (M+H)+; Anal. Calcd for C27H27ClFN2O9: C, 56.21;
H, 4.54; N, 4.86. Found: C, 56.18; H, 4.50; N, 4.90.
DART-HRMS: m/z calcd for
C
28H30N2ClO10 (M+H)+ 589.1589;
found: 589.1582; ESIMS: m/z 589 (M+H)+; Anal. Calcd for
C28H29ClN2O10: C, 57.10; H, 4.96; N, 4.76. Found: C, 57.10; H,
4.96; N, 4.76.
4.1.3.5. 2-Chloro-3-cyano-4-(30,40-dimethoxyphenyl)-6-[(200,300,-
4.1.3.2.
tetra-O-acetyl-b-
2-Chloro-4-(40-bromophenyl)-3-cyano-6-[(200,300,400,600-
-glucopyranosyl)methyl]pyridine (11b). Com-
400,600-tetra-O-acetyl-b-
D-glucopyranosyl)methyl]pyridine (11e).
D
Compound 11e was obtained by the reaction of compound 6e
(0.50 g, 0.83 mmol), POCl3 (10 mL), and toluene (10 mL) as a
white solid (0.35 g, yield 68%); mp 130–132 °C; Rf 0.5 (2.5:97.5,
pound 11b was obtained by the reaction of compound 6b (0.50 g,
0.81 mmol), POCl3 (10 mL), and toluene (10 mL) as a white solid
(0.28 g, yield 77%); mp 140–142 °C; Rf 0.5 (2:98, MeOH–CHCl3);
MeOH–CHCl3); ½a D25
ꢁ
+153 (c 0.1, CHCl3); IR (KBr): mmax 2228
½
a 2D5
ꢁ
ꢀ25.1 (c 0.1, CHCl3); IR (KBr): mmax 2231 (C„N stretching),
(C„N stretching), 1750 cmꢀ1 (O–C@O stretching); 1H NMR
(300 MHz, CDCl3): d 7.28 (s, 1H, ArH), 7.24–7.20 (m, 2H, ArH),
7.01 (d, 1H, J = 8.2 Hz, ArH), 5.24 (t, 1H, J = 9.36 Hz, CH), 5.07 (t,
1H, J = 9.72 Hz, CH), 4.99 (t, 1H, J = 9.63 Hz, CH), 4.22 (dd, 1H,
J1 = 5.16 Hz, J2 = 12.33 Hz, H-6a00), 4.04–4.02 (m, 2H, H-100, H-
6b00), 3.97 (s, 3H, OCH3), 3.96 (s, 3H, OCH3), 3.65–3.59 (m, 1H,
H-500), 3.04–2.99 (m, 2H, CH2), 2.04, 2.03, 2.0, 1.89 (12H,
4 ꢂ OCOCH3); DART-HRMS: m/z calcd for C29H32ClN2O11 (M+H)+
1751 cmꢀ1 (O–C@O stretching); 1H NMR (300 MHz, CDCl3): d
7.71 (d, 1H, J = 8.4 Hz, ArH), 7.61–7.49 (m, 3H, ArH), 7.30–7.27
(m, 1H, ArH), 5.20 (t, 1H, J = 9.63 Hz, CH), 5.04 (t, 1H, J = 9.71 Hz,
CH), 4.95 (t, 1H, J = 9.63 Hz, CH), 4.13 (dd, 1H, J = 4.77 Hz, H-600a),
4.06–3.99 (m, 2H, H-100, H-600b), 3.50–3.61 (m, 1H, H-500), 3.04–
3.00 (m, 2H, CH2), 2.06, 2.05, 2.03, 2.01 (each singlet, 12H,
4 ꢂ OCOCH3); 13C NMR (50 MHz, CDCl3): d 170.3, 170.2, 169.8,

1200
S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
619.1694; found: 619.1686; ESIMS: m/z 619 (M+H)+ Anal. Calcd
for C29H31ClN2O11: C, 56.27; H, 5.05; N, 4.53. Found: C, 56.23;
H, 5.10; N, 4.50.
NaOMe/MeOH (10 mL, 10 mol %) as a white solid (0.2 g, yield
69%); mp = 159–160 °C; Rf 0.5 (2:8, MeOH–CHCl3); ½a D25
ꢀ121 (c
ꢁ
0.1, MeOH); IR (KBr): mmax 3446 (OH stretching), 2227 cmꢀ1
(C„N stretching); 1H NMR (300 MHz, DMSO-d6): d 7.71–7.61 (m,
2H, ArH), 7.19–7.07 (m, 3H, ArH), 5.12 (br s, 1H, OH), 4.88 (br s,
1H, OH), 4.30 (br s, 1H, OH), 4.00 (br s, 1H, OH), 3.83 (s, 3H,
OCH3), 3.75–3.74 (m, 1H), 3.65–3.49 (m, 2H), 3.27–3.01 (m, 4H),
2.99–2.78 (m, 2H); DART-HRMS: m/z calcd for C20H22ClN2O6
(M+H)+ 421.1166; found: 421.1162; ESIMS: m/z 421 (M+H)+; Anal.
Calcd for C20H21ClN2O6: C, 62.84; H, 5.07; N, 5.64. Found: C, 62.80;
H, 5.09; N, 5.62.
4.1.4. General procedure for synthesis of compounds 12a–12e
To a stirring solution of 3-cyano-4-(aryl)-6-[(200,300,400,600-tetra-
O-acetyl-b-D-glucopyranosyl)methyl]pyridines 11a–11e (0.3 g,
0.52 mmol) in MeOH was added a solution of NaOMe (10 mL,
10 mol %) at 25 °C. The reaction mixture was stirred until the dis-
appearance of the starting material was confirmed (TLC). The reac-
tion mixture was neutralized by 2% aq HCl and concentrated in
vacuum to afford the corresponding desired products 12a–12e as
colorless solids.
4.1.4.5.
2-Chloro-3-cyano-4-(30,40-dimethoxyphenyl)-6-[(b-
D-
glucopyranosyl)methyl]pyridine (12e). Compound 12e was ob-
tained by the reaction of compound 11e (0.40 g, 0.64 mmol) and
NaOMe/MeOH (10 mL, 10 mol %) as a white solid (0.25 g, yield
4.1.4.1. 2-Chloro-3-cyano-4-(40-fluorophenyl)-6-[(b-
anosyl)methyl] pyridine (12a). To a stirring solution of 3-cyano-
4-(4’-fluorophenyl)-6-[(200,300,400,600-tetra-O-acetyl-b-
-glucopyran-
D-glucopyr-
D
86%); mp >240 °C; Rf 0.5 (2:8, MeOH–CHCl3); ½a D25
ꢀ167 (c 0.1,
ꢁ
osyl)methyl]pyridine 11a (0.3 g, 0.52 mmol) in MeOH was added a
solution of NaOMe (10 mL, 10 mol %) at 25 °C. The reaction mixture
was stirred until the disappearance of the starting material (TLC).
The reaction mixture was neutralized by 2% aq HCl and concen-
trated in vacuum to afford the desired product 12a as a colorless
solid (0.18 g, yield 88%); mp 125–127 °C; Rf 0.5 (2:8, MeOH–
MeOH); IR (KBr): mmax 3450 (OH stretching), 2108 cmꢀ1 (C„N
stretching); 1H NMR (300 MHz, CDCl3): d 7.38–7.37 (m, 1H, ArH),
7.27–7.14 (m, 2H, ArH), 7.01–6.95 (m, 2H, ArH), 4.05–4.404 (m,
2H), 3.93–3.91 (m, 7H, 2 ꢂ OCH3, OH), 3.72–3.63 (m, 3H), 3.48–
3.38 (m, 3H), 3.30–3.21 (m, 2H), 2.92–2.89 (m, 1H); DART-HRMS:
m/z calcd for C21H23ClN2O7 (M+H)+ 451.1272; found: 451.1269;
ESIMS: m/z 451 (M+H)+; Anal. Calcd for C21H23ClN2O7: C, 55.94;
H, 5.14; N, 6.21. Found: C, 55.90; H, 5.20; N, 6.25.
CHCl3); ½a 2D5
ꢀ55.50 (c 0.1, MeOH); IR (KBr): mmax 3467–3412
ꢁ
(OH stretching), 2231 cmꢀ1 (C„N stretching); 1H NMR (300 MHz,
D2O): d 7.38–7.27 (m, 2H, ArH), 7.18 (s, 1H, ArH), 6.97–6.90 (m,
2H, ArH), 3.42–3.30 (m, 2H), 3.15–3.03 (m, 3H), 2.98–2.89 (m,
3H), 2.68 (dd, 1H, J = 9.15 Hz, –CHbH–); 13C NMR (50 MHz,
DMSO-d6): d 169.6, 167.7, 159.4, 157.8, 136.7, 136.1 (ArCH),
136.0 (ArCH), 135.7, 128.8, 121.1, 120.2, 111.2 (CN), 85.2, 79.1,
76.1, 67.2 (OCH2), 45.9 (CH2); ESIMS: m/z 409 (M+H)+; Anal. Calcd
for C19H18ClFN2O5: C, 55.82; H, 4.44; N, 6.85. Found: C, 55.79; H,
4.40; N, 6.80.
4.2. Biological assays
4.2.1. Assay of
Purified -glucosidase (Sigma–Aldrich) was used as source for
the enzyme for studying the effect of test compounds on -gluco-
sidase inhibition. All the compounds were analyzed for enzyme
inhibitory activity using p nitrophenyl -glucopyranoside (pNPG,
Sigma) as the substrate. The assay system used consisted of 100
of phosphate buffer (0.67 mM, pH 6.8), 100 L of glutathione, 50
of the substrate pNPG, along with 100
of reaction mixture containing 10.0 L of the test compound. The
control tube contained all but 10 L of DMSO instead of compound.
a
-glucosidase33
a
a
a
-D
l
l
L
L
4.1.4.2. 4-(40-Bromophenyl)-2-chloro-3-cyano-6-[(b-
D-glucopyr-
l
anosyl)methyl]pyridine (12b). Compound 12b was obtained by
the reaction of compound 11b (0.50 g, 0.81 mmol) and NaOMe/
MeOH (10 mL, 10 mol %) as a white solid (0.28 g, yield 77%);
lL of the enzyme in 1.0 mL
l
l
mp = 225–227 °C; Rf 0.5 (2:8, MeOH–CHCl3); ½a D25
ꢁ
ꢀ92.09 (c 0.1,
The change in OD, based on the rate of formation of product using a
spectrophotometer, was observed for 3 min at intervals of 30 s.
MeOH); IR (KBr): mmax 3467 (OH stretching), 2233 cmꢀ1 (C„N
stretching); 1H NMR (300 MHz, D2O): d 7.67–7.44 (m, 2H, ArH),
7.34–7.28 (m, 1H, ArH), 3.77–3.60 (m, 5H), 3.36–3.28 (m, 3H),
3.02–2.99 (m, 1H); DART-HRMS: m/z calcd for C19H20BrClN2O5
(M+2H)+ 470.0244; found: 470.0247; ESIMS: m/z 469 (M+H)+;
Anal. Calcd for C19H18BrClN2O5: C, 48.58; H, 3.86; N, 5.96. Found:
C, 48.54; H, 3.82; N, 5.91.
4.2.2. Assay of glucose-6-phosphatase34,35
The liver of an overnight-fasted Wistar rat was excised, and a
10% homogenate was prepared in 150 mM KCl (w/v) using a Potter
Elvejhem glass homogenizer fitted with a Teflon pestle. The
homogenate was centrifuged at 1000ꢂg for 15 min at 4 °C. The
supernatant was decanted and used as the enzyme source.
The effect of the test compound was studied by pre-incubating
4.1.4.3. 4-(40-Benzyloxyphenyl)-2-chloro-3-cyano-6-[(b-
D-gluco-
pyranosyl)methyl]pyridine (12c). Compound 12c was obtained
by the reaction of compound 11c (0.40 g, 0.6 mmol) and NaOMe/
MeOH (10 mL, 10 mol %) as a white solid (0.2 g, yield 84%); mp
100 lM of the compound in a 1.0-mL reaction system for 10 min
and then determining the residual glucose-6-phosphatase activity
according to the method of Hubscher and West.34 The assay system
contained 0.3 M citrate buffer (pH 6.0), 28 mM EDTA, 14 mM NaF,
200 mM glucose-6-phosphate, and an appropriate amount of
enzyme protein. The mixture was incubated at 37 °C for 30 min
after which time the reaction was stopped by addition of 1.0 mL
of 10% TCA. Estimation of inorganic phosphate (Pi) in the pro-
tein-free supernatant was done according to the method of Taus-
sky and Shorr.35 Glucose-6-phosphatase activity was defined as
>240 °C; Rf 0.5 (2:8, MeOH–CHCl3); ½a D25
ꢀ18.7 (c 0.1, MeOH); IR
ꢁ
(KBr): mmax 3445 (OH stretching), 2230 cmꢀ1 (C„N stretching);
1H NMR (300 MHz, DMSO-d6): d 7.74–7.71 (d, 2H, J = 8.76 Hz,
ArH), 7.65 (s, 1H, ArH), 7.52–7.37 (m, 5H, ArH), 7.24 (d, 2H,
J = 8.76 Hz, ArH), 5.23–5.21 (m, 3H, OCH2, OH), 5.06 (d, 1H,
J = 4.05 Hz, OH), 4.98 (d, 1H, J = 3.99 Hz, OH), 4.43 (t, 1H,
J = 5.76 Hz, OH), 3.59–3.49 (m, 3H), 3.23–3.19 (m, 2H), 3.10–3.01
(m, 3H), 2.90 (dd, 1H, J1 = 9.39 Hz, J2 = 14.79 Hz, –CHbH–); DART-
HRMS: m/z calcd for C26H26ClN2O6 (M+H)+ 497.1479; found:
497.1469; ESIMS: m/z 497 (M+H)+. Anal. Calcd for C26H25ClN2O6:
C, 62.84; H, 5.07; N, 5.64. Found: C, 62.80; H, 5.00; N, 5.68.
lM of Pi release per min per mg protein.
4.2.3. Assay of glycogen phosphorylase35,36
A rat of Wistar strain was put on 16 h fasting and its liver was
excised. A 10% homogenate was prepared in 150 mM KCl (w/v)
using a Potter–Elvejhem glass homogenizer fitted with a Teflon
pestle. The homogenate was centrifuged at 1000ꢂg for 15 min at
4 °C, supernatant was decanted and used as the enzyme source.
4.1.4.4. 2-Chloro-3-cyano-6-[(b-D
-glucopyranosyl)methyl]-4-(40-
methoxyphenyl)pyridine (12d). Compound 12d was obtained
by the reaction of compound 11d (0.40 g, 0.68 mmol) and

S. S. Bisht et al. / Carbohydrate Research 346 (2011) 1191–1201
1201
The effect of the test compound was studied by pre-incubating
a 100 M solution of the test compound in 1.0 mL of the reaction
Seerat and Rahul, respectively. We sincerely acknowledge the
financial assistance from DRDO and CSIR New Delhi.
l
system for 10 min, and then determining the residual glycogen
phosphorylase activity according to the method of Rall et al.36
The assay mixture contained 0.2 mL of mixture A [glycogen
0.057 g, glucose-1-phosphate 0.188 g, NaF 0.042 g and 50 AMP
(4 mM) in 10 mL of distilled water] and 0.1 mL of mixture B, the
enzyme protein. It was incubated at 37 °C for 30 min, after which
time the reaction was stopped by addition of 0.1 mL of 10% TCA,
and then 0.4 mL of NaOAc (100 mM) was added to prevent the
spontaneous hydrolysis of G-1-P present in the reaction mixture.
The estimation of inorganic phosphate in the protein-free superna-
tant was done according to the method of Taussky and Shorr.35
Supplementary data
Supplementary data (ESMS, HRMS, 1H, 13C NMR, HSQC and
HMBC spectra of prototype compounds) associated with this arti-
cle can be found, in the online version, at doi:10.1016/j.carres.
2011.03.006.
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4.2.4.3. Grouping and treatment. The diabetic animals were di-
vided into groups with five animals in each group and were admin-
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acacia at desired dose levels (100 mg/kg) in the case of synthetic
compounds and 100 mg/kg in case of the standard drug metfor-
min). Animals of the control group were given an equal amount
of 1% gum acacia and termed as sham controls. A sucrose load of
2.5 g/kg was given to each animal orally exactly after 30 min post
administration of the test sample/vehicle. Blood glucose profile of
animals of all groups was again checked at 30, 60, 90, 120, 180,
240, 300, and1440 min post administration of sucrose. Food but
not water was withheld from the cages during the course of
experimentation.
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4.2.4.4. Statistical analysis. Quantitative glucose tolerance of
each animal was calculated by the area under the curve (AUC)
method using Prism Software. The area under the curve of the con-
trol group and the experimental group was compared to determine
the percent antihyperglycemic activity. Statistical comparisons
were performed by one-way analysis of variance (ANOVA), fol-
lowed by Dunnett’s multiple range test (DMRT). The results were
considered statistically significant if the ‘p’ values were 0.05 or less.
Results were expressed as mean SEM. Samples showing signifi-
cant inhibition (p <0.05, p <0.01, and p <0.001) on postprandial
hyperglycemia (AUC) were considered as active samples.
Acknowledgments
This is CDRI Communication No. 8025. The authors thank CSIR
and DRDO New Delhi for the SRF and PA to Surendra, Anindra,