Structure-activity relationship studies of 4-benzyl-1H-pyrazol-3-yl β-d-glucopyranoside derivatives as potent and selective sodium glucose co-transporter 1 (SGLT1) inhibitors with therapeutic activity on postprandial hyperglycemia

Article abstract of DOI:10.1016/j.bmc.2012.09.037

Sodium glucose co-transporter 1 (SGLT1) plays a dominant role in the absorption of glucose in the gut and is considered a promising target in the development of treatments for postprandial hyperglycemia. A series of 4-benzyl-1H-pyrazol-3-yl β-d-glucopyran

Full text of DOI:10.1016/j.bmc.2012.09.037

Bioorganic & Medicinal Chemistry 20 (2012) 6598–6612
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Structure–activity relationship studies of 4-benzyl-1H-pyrazol-3-yl
b-D-glucopyranoside derivatives as potent and selective sodium glucose
co-transporter 1 (SGLT1) inhibitors with therapeutic activity on postprandial
hyperglycemia
Nobuhiko Fushimi a, , Hideki Fujikura a, Hiroaki Shiohara a, Hirotaka Teranishi a, Kazuo Shimizu a,
⇑
Shigeru Yonekubo a, Kohsuke Ohno a, Takashi Miyagi a, Fumiaki Itoh a, Toshihide Shibazaki a,
Masaki Tomae a, Yukiko Ishikawa-Takemura a, Takeshi Nakabayashi a, Noboru Kamada a,
Tomonaga Ozawa a, Susumu Kobayashi b, Masayuki Isaji a
a Central Research Laboratory, Kissei Pharmaceutical Company, 4365-1 Kashiwabara, Hotaka, Azumino, Nagano Prefecture 399-8304, Japan
b Faculty of Pharmaceutical Sciences, Tokyo University of Science (RIKADAI), 2641 Yamazaki, Noda-shi, Chiba Prefecture 278-8510, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
Sodium glucose co-transporter 1 (SGLT1) plays a dominant role in the absorption of glucose in the gut
and is considered a promising target in the development of treatments for postprandial hyperglycemia.
A series of 4-benzyl-1H-pyrazol-3-yl b-D-glucopyranoside derivatives have been synthesized, and its
inhibitory activity toward SGLTs has been evaluated. By altering the substitution groups at the 5-position
of the pyrazole ring, and every position of the phenyl ring, we studied the structure–activity relationship
(SAR) profiles and identified a series of potent and selective SGLT1 inhibitors. Representative derivatives
showed a dose-dependent suppressing effect on the escalation of blood glucose levels in oral mixed car-
bohydrate tolerance tests (OCTT) in streptozotocin–nicotinamide-induced diabetic rats (NA-STZ rats).
Ó 2012 Elsevier Ltd. All rights reserved.
Received 13 August 2012
Revised 12 September 2012
Accepted 13 September 2012
Available online 24 September 2012
Keywords:
Diabetes
Postprandial hyperglycemia
SGLT1
SGLT1 inhibitor
1. Introduction
(NIDDM) in impaired glucose tolerance (IGT) patients (STOP-NID-
DM), the a-glucosidase inhibitor, acarbose, not only decreased the
Diabetes mellitus (DM) is a group of metabolic diseases charac-
terized by hyperglycemia resulting from defects in insulin secretion
and/or insulin action.1 The global prevalence of DM is increasing; in
2011, there were 366 million people with DM, and this number is
expected to rise to 552 million by 2030.2 DM is a risk factor for
microvascular diseases including retinopathy, neuropathy, and
nephropathy and is also known to be associated with increases in
cardiovascular (CV) disease and mortality. Increased postprandial
glucose excursion is a frequently observed abnormality in DM that
is associated with endothelial dysfunction as well as oxidative and
nitrosative stress. A large number of epidemiological studies have
consistently shown that postprandial hyperglycemia is a powerful
predictor of CV disease. Hence, there is now greater awareness of
progression to DM but also protected them against CV disease.4
Carbohydrates in the diet, mainly in the form of poly- or disac-
charides, are digested into monosaccharides by hydrolytic en-
zymes (a-amylase, a-glucosidases, and b-galactosidases) before
absorption.5 The resulting monosaccharides are absorbed via sev-
eral types of transporters in the small intestine.6 Sodium glucose
co-transporter 1 (SGLT1), expressed in the brush border membrane
of the enterocytes, plays a central role in the absorption of glucose
and galactose.7 Mutations in the SGLT1 gene cause malabsorption
of these sugars.8 Furthermore, SGLT1 protein levels were found
to be higher in duodenum biopsy samples obtained from diabetic
patients relative to the control subjects.9 Increased expression of
SGLT1 in the small intestine was also observed in streptozotocin-
induced diabetic rats.10 These findings suggest that the inhibition
of SGLT1 can be an efficacious approach to improve postprandial
hyperglycemia in DM and IGT patients.
the importance of controlling postprandial hyperglycemia.3
a-Glu-
cosidase inhibitors that delay the digestion of carbohydrates have
been used clinically for the treatment of postprandial hyperglyce-
mia. In a clinical trial to prevent non-insulin-dependent DM
To date, multiple classes of compounds have been reported to
show inhibitory activities on SGLTs.11 However, most of these com-
pounds intend to inhibit renal SGLT2, and little is known about the
structure–activity relationship (SAR) and the in vivo efficacy of
⇑
Corresponding author. Tel.: +81 263 82 8820; fax: +81 263 82 8827.
E-mail address: nobuhiko_fushimi@pharm.kissei.co.jp (N. Fushimi).
0968-0896/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmc.2012.09.037

N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
6599
that of naturally occurring phlorizin (6), a nonselective inhibitor
of these SGLTs.17 With this novel lead compound in hand, we em-
barked on a research program to improve the potential of SGLT1
inhibitors on postprandial hyperglycemia and performed a series
of SAR studies focusing on SGLT1 activity to develop improved pro-
file compounds. Since compounds that inhibit both SGLTs could in-
duce urinary glucose excretion and normalize blood glucose levels
by inhibiting renal SGLT2 if they were absorbed, our exploratory
effort was directed to achieve an SGLT1-selective molecule to
investigate efficacy based on the inhibition of SGLT1 alone. Ulti-
mately, this selectivity for SGLT1 activity on intestinal transporters
might facilitate the combined use with SGLT2 inhibitors targeting
renal glucose excretion.
H
CF3
O
O
N
N
N
N
O O
R
O
O
R
O
O
O
HO
HO
O
HO
OH
OH
HO
OH
OH
OH
OH
5 : lead compound
3 : R = CO2Et
1 : R = CO2Et
sergliflozin
2 : R = H
remogliflozin etabonate
4 : R = H
remogliflozin
sergliflozin-A
OH
O
OH
HO
Cl
O
O
O
O
HO
HO
In the present paper, we report the synthesis and SAR of 4-ben-
zyl-1H-pyrazol-3-yl b-D-glucopyranoside derivatives to find potent
HO
OH
HO
OH
OH
OH
and selective SGLT1 inhibitors. We also describe the in vivo phar-
macological profiles of several derivatives in association with their
in vitro intestinal metabolic stability and aqueous solubility.
7 : dapagliflozin
6 : phlorizin
Figure 1. Structure of known SGLT inhibitors and the lead compound in this study.
2. Chemistry
SGLT1-targeted inhibitors.12 SGLT2, the isoform of SGLT1, is dis-
tributed predominantly in the proximal tubules of the kidney,
and plays a dominant role in the reabsorption of glomerular-fil-
tered glucose.13 Since SGLT2 inhibitors can normalize blood glu-
cose levels by excreting excess glucose into the urine, large
numbers of compounds developed by pharmaceutical companies
are undergoing clinical trials to evaluate their efficacy as antidia-
betic agents.14 We previously discovered two prodrugs of selective
SGLT2 inhibitors, sergliflozin (1)15 and remogliflozin etabonate
(3)16 (Fig. 1). In the course of this research, 4-benzyl-5-trifluoro-
4-Benzyl-1H-pyrazol-3-yl b-D-glucopyranoside derivatives were
prepared as illustrated in Scheme 1. Benzyl halides or benzyl alco-
hols, in which hydroxyl groups of benzyl alcohols were methane-
sulfonylated using methanesulfonyl chloride (MsCl) with Et3N
before condensation, were allowed to react with ketoesters in the
presence of NaH to afford 2-substituted ketoesters (8a–y). Subse-
quent cyclization with hydrazine monohydrate gave aglycones
(9a–y). Reaction of aglycones (9a–y) with acetobromoglucose was
carried out in two-phase solvents consisting of CH2Cl2 and aqueous
NaOH solution in the presence of the phase-transfer catalyst,
BnN(n-Bu)3Cl.18 The glucosidated structure of the product was con-
firmed by X-ray crystal structure analysis of the intermediate (10a),
methyl-1H-pyrazol-3-yl b-D-glucopyranoside (5), an analog of
remogliflozin (4), was found to have strong inhibitory activity on
both SGLTs, and its potency toward SGLT1 was comparable with
a (for X = OH)
H
R1
R4
R3
R4
R3
R4
R3
O
O
R1
N
c
d
b
HN
X
RO
O
R2
R2
R2
8a y
9a y
X = Cl, Br, OH
R = Me, Et
H
R1
H
R1
R4
R3
R4
R3
N
N
N
N
e
11a c, f, t, w, y
12a g
R2
R2
O O
O
O
f
HO
HO
AcO
AcO
OH
OAc
OAc
OH
10a y
11a y
Cpd
8–11
Cpd
R1
Cpd
R1
R2
R3
R4
R2
R3
R4
R1
R2 R3
R4
8–11
12
a
b
c
a
b
c
d
e
f
CF3
CF3
H
H
H
OBn
H
OBn
H
n
o
p
q
r
iPr OMe
iPr CN
H
H
H
CF3
CF3
H
H
H
OH
H
OH
H
H
F
CF3 OBn
Me OBn
Et OBn
iPr OBn
tBu OBn
cPr OBn
H
iPr
iPr
iPr
iPr
iPr
iPr
iPr
iPr
H
H
H
H
H
H
H
H
H
CF3 OH
iPr OH
H
H
H
H
Me
Et
d
e
H
H
H
H
H
iPr
iPr
H
H
H
OH
H
H
H
s
H
OMe
OBn
CN
H
f
OH
H
g
h
i
H
H
t
H
g
iPr Me
OH
H
H
u
v
w
x
y
H
iPr
iPr
iPr
H
F
H
H
Me
OBn
H
j
H
H
H
k
l
Me
H
H
iPr Me
iPr Me
Me
OBn
iPr CF3
iPr Et
H
H
H
m
H
H
Scheme 1. Reagents and conditions: (a) MsCl, Et3N, DME or THF, 0 °C; (b) NaH, R1COCH2CO2R, DME or THF, 0 °C to reflux; (c) N2H4ꢀH2O, toluene or EtOH, reflux or rt; (d) 5 M
NaOH aq, BnN(n-Bu)3Cl, acetobromoglucose, CH2Cl2, rt; (e) NaOMe, MeOH, rt; (f) H2, Pd–C, MeOH, rt.

6600
N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
Figure 2. ORTEP representation of the X-ray crystal structure of 10a.
as shown in Figure 2. Finally, the acetyl groups protecting the hy-
droxyl groups on the sugars were removed by hydrolysis using
NaOMe in MeOH to give 4-benzyl-1H-pyrazol-3-yl b-D-glucopyran-
oside derivatives 11a–y. For the synthesis of compounds possessing
hydroxyl groups on the aglycones (12a–g), benzyl protective
groups were removed by Pd–C-catalyzed hydrogenation.
clinic,20 with a hydroxyl group enhanced the inhibitory activity
of SGLT1 and reduced the selectivity for SGLT2.21 These findings
led us to assume that the introduction of hydroxyl groups into
the distal phenyl ring might also improve SGLT1 inhibitory activity
in our scaffold. Hence, we examined the effects of introducing hy-
droxyl groups into the phenyl ring of our lead compound (5). Since
the substitution point might slide between each scaffold, we exam-
ined every position of the phenyl ring of compound 5 to confirm
suitability, and the results are shown in Table 1. Contrary to our
expectation, none of the hydoxylated derivatives (12a–c) showed
improved inhibitory activity for SGLT1. Indeed, of these com-
pounds, only the 4-hydroxylated compound (12a) showed a main-
tained, but not enhanced, activity with slightly diminished
selectivity for SGLT1 compared with the unsubstituted compound
5. Introduction of hydroxyl groups into other positions decreased
the activity for both SGLTs (12b, c). Although no improvements
were observed in these initial attempts, the 4-position was rela-
tively favored for hydroxyl-group substitution, indicating that this
position on a benzyl pyrazole derivative might be similar to that in
the other scaffolds described above. During these explorations,
synthetic precursors possessing benzyl protective groups (11a–c)
were also evaluated for their activities. Substitution at the 3-and
3. Results and discussion
All compounds were screened for their inhibitory effects on the
uptake of radiolabeled
a-methyl-D
-glucopyranoside ([14C]-AMG)
into COS-7 cells transiently expressing human SGLT1 (hSGLT1) or
SGLT2 (hSGLT2).
In phrolizin (6), one of the hydroxyl groups exists in the 4-posi-
tion of the distal phenyl ring. SAR studies of 40-dehydroxyphlorizin
derivatives demonstrated that a hydroxyl group at this position
helps to retain the inhibitory effects on glucose uptake in the small
intestine.19 It has also been reported that the replacement of the
ethoxy group at the 4-position of the distal phenyl ring of dapagli-
flozin (7), the most advanced selective SGLT2 inhibitor in the
Table 1
The hSGLT1 and hSGLT2 inhibitory activities and selectivity of 5-trifluoromethyl
pyrazole derivatives substituted with hydroxyl and benzyloxy groups on the phenyl ring
Table 2
The hSGLT1 and hSGLT2 inhibitory activities and selectivity of 4-(2-benzyloxy
phenylmethyl)pyrazole derivatives substituted with various groups on the 5-position
of the pyrazole ring
H
CF3
4
3
N
R
N
2
O
O
H
R1
HO
HO
N
N
OH
OH
O
O
O
HO
HO
a
Compound
R
SGLT IC50 (nM)
Selectivityb
OH
OH
hSGLT1
hSGLT2
536 19c
442
500
1940
4310
1720
8930
37
38 2c
342
154
2790
14,900
7
5680
14,000
0.071
0.77
0.31
1.4
Compound
R1
SGLT IC50a (nM)
Selectivityb
6
5
—
H
hSGLT1
hSGLT2
12a
12b
12c
11a
11b
11c
4-OH
3-OH
2-OH
4-OBn
3-OBn
2-OBn
11c
11d
11e
11f
11g
11h
CF3
Me
Et
iPr
tBu
cPr
37
14,000
5010
13,900
74,000
64,700
156,000
380
5.8
45
1200
320
560
871
306
60
203
278
3.5
0.0041
0.64
380
a
b
c
IC50 values for SGLT inhibition were determined in duplicate or triplicate.
Selectivity values were calculated by IC50 hSGLT2/IC50 hSGLT1.
Mean S.E. of 19 experiments.
a
IC50 values for SGLT inhibition were determined in duplicate or triplicate.
Selectivity values were calculated by IC50 hSGLT2/IC50 hSGLT1.
b

N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
6601
Table 4
Vehicle
The hSGLT1 and hSGLT2 inhibitory activities and selectivity of 5-isopropylpyrazole
derivatives substituted with various groups on the phenyl ring
3 mg/kg
400
300
200
100
0
10 mg/kg
30 mg/kg
H
R4
N
N
R3
R2
O
O
HO
HO
OH
OH
Compound
R2
R3
R4
SGLT IC50a (nM)
Selectivityb
hSGLT1
hSGLT2
11i
H
Me
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
F
Me
Et
OMe
OH
CN
H
H
Me
OH
738
725
1190
94
223
689
207
4310
1330
1230
246
320
2020
18,500
4140
231
68
377
2.7
26
11k
11p
11q
11r
11s
12e
11u
11v
12f
11x
12g
3.5
2.5
0.30
0.55
1.4
0.83
4.5
11
285
3580
5980
13,300
6010
6890
Me
OH
H
H
Me
Me
24
22
H
0
1
2
3
a
IC50 values for SGLT inhibition were determined in triplicate.
Selectivity values were calculated by IC50 hSGLT2/IC50 hSGLT1.
Time after administration (h)
b
Figure 3. Effects of compound 11f on OCTT in NA-STZ rats. Points and bars
represent the mean and S.E. values, respectively (n = 7).
(IC50 >100 lM), it was expected that 11c would show weak efficacy
in vivo. To achieve efficacious compounds in vivo, the metabolic
stability of the glucosidic linkages should be improved. As common
structural features between phlorizin (6) and compound 11c,
electron-withdrawing groups on the ring connecting to glucose
were suggested. We anticipated that the trifluoromethyl group
may have a deleterious effect on the stability of the glucosidic
linkages, and turned our attention to find other non-electron-
withdrawing groups at the 5-position of the pyrazole ring of
compound 11c.
Table 3
The hSGLT1 and hSGLT2 inhibitory activities and selectivity of 5-isopropylpyrazole
derivatives substituted with various groups on the 2-position of the phenyl ring
H
N
N
R2
O
O
HO
HO
OH
OH
Compound
R2
SGLT IC50a (nM)
Selectivityb
Table 2 shows the results of investigation of substituents at the
5-position of the pyrazole ring. Replacement with a methyl group
markedly decreased the inhibitory activity and selectivity for
SGLT1 (11d), while introducing longer alkyl groups (11e–h) en-
hanced activity and selectivity compared with compound 11d. Of
these, the isopropyl derivative 11f showed comparable potency
and further improved the selectivity for SGLT1 relative to com-
pound 11c. The bulkier tert-butyl group (11g) and cyclized propyl
group (11h) showed diminished activity on SGLT1 compared with
the isopropyl group (11f), indicating that SGLT1 would recognize
subtle difference in the shape of the groups incorporated onto
the 5-position of the pyrazole ring. On the basis of these results,
the isopropyl group was considered to be optimal as a substitution
group of this position, and an intestinal metabolic stability test of
compound 11f was performed (Table 5). Fortunately, 11f clearly
improved stability compared with the 5-trifluoromethyl pyrazole
derivative (11c), and 51% of 11f was found in the unchanged form,
even at 60 min. These findings demonstrate that substituents at
the 5-position of the pyrazole ring would affect not only the activ-
ity and selectivity for SGLT1 but also the metabolic stability of the
glucosidic linkage connecting to the 3-position. Prior to in vivo
examinations, the inhibitory activity on rat SGLTs (rSGLT1,2) was
also evaluated (Table 5). Compound 11f showed decreased activity,
especially in rSGLT1 relative to hSGLT1, but was still effective com-
pared with phlorizin (6), and high selectivity remained.
hSGLT1
hSGLT2
11f
11i
11j
11k
11l
11m
11n
12d
11o
OBn
H
F
Me
CF3
Et
OMe
OH
CN
60
738
298
725
2430
675
7280
9520
7170
74,000
2020
1390
18,500
60,500
14,100
204,000
75,900
65,700
1200
2.7
4.7
26
25
21
28
8.0
9.2
a
IC50 values for SGLT inhibition were determined in duplicate or triplicate.
Selectivity values were calculated by IC50 hSGLT2/IC50 hSGLT1.
b
4-positions resulted in loss of activity (11a, b); however the 2-ben-
zyloxy-substituted derivative (11c) showed greater selectivity
against SGLT2 with dramatically improved the inhibitory activity
on SGLT1 compared with the unsubstituted compound 5.
In general, to find selective SGLT2 inhibitors, the exploratory
efforts on the substituents of the distal phenyl rings were mainly
directed to the 4-position. Hence, we found that this remarkable
substituent effect of the 2-benzyloxy group was unique, and this
encouraged us to explore derivatives of this scaffold further. Before
another investigation, we assessed the intestinal metabolic stabil-
ity of the compound 11c. It is known that phlorizin (6) is easily
hydrolyzed into its aglycone form, phloretin, by b-glucosidase in
the intestine.17 As shown in Table 5, incubation with rat intestinal
microsomes immediately decreased compound 11c, as well as
phlorizin (6), and the corresponding aglycone of 11c was detected.
Since, the aglycone of 11c (9c) has no inhibitory activity on SGLT1
Despite the strong inhibitory activity on SGLT1 and the rela-
tively stable profile in the intestine, compound 11f showed insuf-
ficient efficacy in an oral mixed carbohydrate tolerance test
(OCTT) in streptozotocin–nicotinamide-induced diabetic rats
(NA-STZ rats).22 As shown in Figure 3, suppressing effects of com-

6602
N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
Table 5
Aqueous solubility and parameters in rats: inhibitory activity and selectivity of SGLT1 and SGLT2, intestinal metabolic stability, and urinary glucose excretion of key compounds
a
Compound
SGLT IC50 (nM)
Selectivityb
Metabolic stability at 30/60 minc
Solubilityd (mg/mL)
Urinary excreted glucosee (mg)
rSGLT1
rSGLT2
Test compound
Sergliflozin (1)
Control
6
350 34f
227
96 11f
40,800
100,000
NTh
4650
4370
0.27
180
320
—
69
24
ND/NDg
ND/NDg
71/51
NTh
91/67
96/83
NTh
NTh
0.24
19
3.7
>18
NTh
—
—
—
—
11c
11f
11i
11x
12g
NTh
308
0.4 0.1
64.1 5.0***
—
0.4 0.1
—
0.5 0.1
0.5 0.1
NTh
NTh
67
183
0.8 0.1
0.7 0.2
53.3 8.2**
53.3 8.2**
a
b
c
IC50 values for SGLT inhibition were determined in duplicate or triplicate.
Selectivity values were calculated by IC50 rSGLT2/IC50 rSGLT1.
Incubated with rat intestinal microsomes. Data are shown as % remaining.
Determined in distilled water.
Each value represents the mean S.E. (n = 5).
p < 0.01 versus control.
p <0.001 versus control.
Mean S.E. of three experiments.
ND = not detected.
NT = not tested.
d
e
**
***
f
g
h
Vehicle
Vehicle
400
1 mg/kg
0.3 mg/kg
1 mg/kg
3 mg/kg
3 mg/kg
300
10 mg/kg
300
200
*
200
100
0
***
***
***
**
**
**
***
100
0
0
1
2
3
0
1
2
3
Time after administration (h)
Time after administration (h)
Figure 5. Effects of compound 12g on OCTT in NA-STZ rats. Points and bars
represent the mean and S.E. values, respectively (n = 7). Asterisks indicate signif-
icant differences from the control (⁄p <0.05, ⁄⁄p <0.01).
Figure 4. Effects of compound 11x on OCTT in NA-STZ rats. Points and bars
represent the mean and S.E. values, respectively (n = 7). Asterisks indicate signif-
icant differences from the control (⁄⁄p <0.01, ⁄⁄⁄p <0.001).
the 2-position of the phenyl ring, and the results are illustrated
in Table 3.
pound 11f on the escalation of blood glucose was actually observed
at 30 min, although the effect was moderate and could not be ob-
served after 60 min. Moreover, no dose-dependent efficacy was ob-
served. Subsequent assessment of the aqueous solubility of
compound 11f (Table 5) suggested that low solubility to water
might explain this unsatisfactory result. Since an SGLT1 inhibitor
might compete against highly water-soluble glucose in the intes-
tine, higher water solubility might be advantageous in order to
facilitate rapid diffusion into the gut and efficient inhibition of
SGLT1. To confirm this idea, we next directed our attention to find
highly soluble compounds.
Introduction of a fluorine atom enhanced activity compared
with the unsubstituted compound 11i, but the improvement in
selectivity was slight (11j). Although the inhibitory activity for
SGLT1 was not improved, a moderate recovery of the selectivity
for SGLT1 was observed after a methyl group was introduced
(11k). Replacement of this methyl group with a trifluoromethyl
group resulted in marked loss of activity (11l). Elongation into an
ethyl group led to almost no improvement in activity, but slightly
reduction in selectivity versus SGLT2 (11m). Compounds contain-
ing a methoxy group (11n) demonstrated weaker activity com-
The aromatic ring count of a molecule must be reduced to im-
prove its aqueous solubility.23 Removal of the 2-benzyloxy group
on the phenyl ring of compound 11f dramatically increased
aqueous solubility (11i), as expected (Table 5). However, the
unsubstituted compound 11i showed markedly decreased inhib-
itory activity and selectivity for SGLT1. To recover these parame-
ters, other less hydrophobic groups were next introduced into
pared with compounds containing a benzyloxy group (11f),
suggesting that the phenyl ring on the methoxy group plays a crit-
ical role in improving SGLT1 inhibitory activity in compound 11f.
In addition, the introduction of either OH (12d) or CN (11o) re-
duced potency, indicating that these small polar groups were not
preferred at this position. In these explorations of less hydrophobic
substitution groups at the 2-position of phenyl ring, we could not

N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
6603
find groups that were fully comparable to the benzyloxy group.
However, it was found that the selectivity toward SGLT1 could at
least be recovered moderately by the introduction of a methyl
group (11k).
the excretion of glucose into the urine. These findings strongly sug-
gest that the effects of 11x and 12g in suppressing the escalation of
blood glucose levels in OCTT in NA-STZ rats were based not on the
enhancement of urinary glucose excretion by the inhibition of
SGLTs in the renal proximal tubule of the kidney but on slowing
or preventing digestive glucose absorption by SGLT1 inhibition in
the small intestine.
To investigate whether further improvement of the profile
could be achieved, we next studied the effects of substituents at
the 4-position of the phenyl ring, and the results are shown in
Table 4. Unlike at the 2-position, introduction of a fluorine atom
at the 4-position resulted in almost no improvement of inhibitory
activity and selectivity for SGLT1 (11p) compared with unsubsti-
tuted compound 11i. While, SGLT2 inhibition was also enhanced
and selectivity was not improved, introduction of a methyl group
was effective in improving inhibitory activity on SGLT1 (11q).
Extension of this methyl group into an ethyl group (11r) or
replacement with methoxy group (11s) both reduced SGLT1 inhib-
itory activity with loss of selectivity versus SGLT2, suggesting that
these groups were relatively SGLT2-targeted. Compared with the
4-methyl derivative 11q, the 4-hydroxylated compound 12e was
not potent and was slightly less selective toward SGLT1, but
showed an enhancement of the inhibition of SGLT1 from the
unsubstituted compound 11i. This finding was not seen in the 5-
trifluoromethyl pyrazole derivative (5 vs 12a), indicating that sub-
stitution of groups at the 5-position of the pyrazole ring might
influence the alignment of the distal phenyl ring and groups substi-
tuted onto this phenyl ring. A cyano group was also examined as
another hydrophilic group, resulting in marked decrease in activity
(11u). The relatively preferred methyl and hydroxyl groups were
also introduced into the 3-position of the phenyl ring. A slight-
to-moderate improvement in selectivity was observed, but the
inhibitory activity on SGLT1 was reduced compared with that of
the unsubstituted compound 11i (11v, 12f), indicating that the
3-position was not favored for the incorporation of these groups
compared with other positions of the phenyl ring.
On the basis of these SAR studies, the effects of combining the rel-
atively preferred substitution groups of the phenyl ring were finally
investigated. A 2-methyl group was introduced as a selectivity-
improving group into compounds bearing a 4-methyl or 4-hydroxyl
group. Although the inhibition of SGLT1 was slightly reduced, the
resulting disubstituted compounds 11x and 12g showed better
selectivity profiles compared with basal monosubstituted com-
pounds 11q and 12e. Furthermore, since these non-aromatic func-
tional groups would have less effect on the hydrophobicity of
molecules, the aqueous solubility of compounds 11x and 12g was
higher than that of the compound possessing a 2-benzyloxy group
(11f) (Table 5).
4. Conclusions
A
series of 4-benzyl-1H-pyrazole-3-yl b-D-glucopyranoside
derivatives was explored to develop a compound with potent
and selective SGLT1 inhibitory activity. These investigations iden-
tified that substituents at the 5- and 2-positions of the pyrazole
and phenyl rings, respectively, could improve both inhibitory
activity and selectivity toward SGLT1, while substitution at the
4-position on the phenyl ring could enhance only inhibitory activ-
ity. In vitro intestinal metabolic stability and aqueous solubility of
several compounds were also evaluated, and groups at the 5-posi-
tion of the pyrazole ring was found to affect the hydrolytic stability
of the glucosidic linkage, and compounds possessing non-aromatic
functional groups on the phenyl ring showed higher solubility.
These SAR studies identified compounds 11x and 12g as potent
and selective SGLT1 inhibitors, both of which showed improved
in vitro intestinal stability over phlorizin (6), high solubility to
water, and robust efficacy in OCTT in NA-STZ rats. To the best of
our knowledge, this is the first report of SAR studies of SGLT1-
selective inhibitors. We continue the optimization and evaluation
of these derivatives and will report further developments in due
course.
5. Experimental
5.1. Chemistry
1H NMR spectra were recorded using Bruker Avance II+ 400
instruments. 13C NMR spectra were recorded using Bruker Avance
II+ 400 or Avance III 600 instruments, and chemical shifts were re-
ported in parts per million (d) downfield from tetramethylsilane as
the internal standard. Peak patterns were shown using the follow-
ing abbreviations: br, broad; s, singlet; d, doublet; t, triplet; q,
quartet; and m, multiplet. Mass spectra (MS) were measured using
an Agilent Technologies 6140 (ESI) or Shimadzu LCMS-2010EV
(ESI). High resolution mass spectra (HRMS) were recorded using
an Agilent Technologies 6520 Accurate-Mass Q-TOF instrument.
Silica gel 60F254-precoated glass plates from Merck KgaA were
used for thin layer chromatography (TLC). Flash or medium pres-
sure liquid chromatography (MPLC) was performed on silica gel
BW-350 from Fuji Silysia Chemical Ltd or SNAP cartridge KP-Sil
from BiotageÒ. All reagents and solvents were commercially avail-
able unless otherwise indicated.
As shown in Figures 4 and 5, in contrast to the results of the
less water-soluble compound 11f, the lowering of blood glucose
levels by compounds 11x and 12g, respectively, in OCTT in NA-
STZ rats was markedly improved in a dose-dependent manner.
In this model, metabolically-unstable phlorizin (6, 1 mg/kg)
showed almost no efficacy (data not shown). These findings sug-
gest that both improved metabolic stability in intestinal content
and higher aqueous solubility would contribute to the impressive
in vivo efficacy of compounds 11x and 12g. As common adverse
5.1.1. 4-(4-Benzyloxyphenylmethyl)-5-trifluoromethyl-1,2-
dihydro-3H-pyrazol-3-one (9a)
effects with a
-glucosidase inhibitors,4 these compounds also may
MsCl (0.88 mL, 11.4 mmol), under ice cooling, was added to a
solution of 4-benzyloxyphenylmethanol (2.33 g, 10.9 mmol) and
Et3N (1.67 mL, 11.9 mmol) in 1,2-dimethoxyethane (DME:
10 mL), and the mixture was stirred for 5 min. The insoluble mate-
rial was removed by filtration and washed with DME (2 mL). The
filtrate and washing were added to the mixture prepared by the
addition of NaH (478 mg, 12.0 mmol, 60% oil dispersion) into a
solution of ethyl 4,4,4-trifluoroacetoacetate (2.00 g, 10.9 mmol)
in DME (15 mL) under ice cooling and stirring for 5 min, then the
mixture was refluxed for 11 h. After cooling to room temperature,
1 M HCl (20 mL) was added, and the resulting mixture was
cause gastrointestinal disturbances. However, the obvious symp-
toms such as soft feces or diarrhea were not observed in this
study.
In a pharmacokinetic study of compound 12g (10 mg/kg) in
Sprague–Dawley rats, very low levels (8.2 ng/mL) of 12g were de-
tected in the plasma and were eliminated immediately, causing no
detection at 40 min after administration (data not shown). Further-
more, urinary glucose excretion was not observed in normal rats
when 11x and 12g were administered orally at a dose of 10 mg/
kg, and urine was collected overnight (Table 5). In contrast, the or-
ally available SGLT2 inhibitor sergliflozin significantly increased

6604
N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
extracted with EtOAc. The extract was washed with brine and
dried over anhydrous MgSO4. The solvent was removed under re-
duced pressure to give crude ethyl 2-(4-benzyloxyphenylmeth-
yl)-4,4,4-trifluoroacetoacetate (8a). This material was dissolved
in toluene (30 mL), and N2H4ꢀH2O (1.05 mL, 21.7 mmol) was added
to the solution. After refluxing for 7 h, water was added, and the
mixture was extracted with EtOAc. The extract was washed with
brine and dried over anhydrous MgSO4. Next, the solvent was re-
moved under reduced pressure, and the residue was purified by
column chromatography on silica gel (eluent: CH2Cl2/
MeOH = 10:1) to give 9a (2.46 g, 65%) as a white solid.
5.1.7. 4-(2-Benzyloxyphenylmethyl)-5-tert-butyl-1,2-dihydro-
3H-pyrazol-3-one (9g)
The title compound was prepared from 2-benzyloxyphenyl-
methanol and methyl 4,4-dimethyl-3-oxopentanoate, as described
for the synthesis of 9a, in 3% yield.
1H NMR (DMSO-d6) d: 1.13 (9H, s), 3.71 (2H, s), 5.17 (2H, s),
6.74–6.83 (2H, m), 7.01 (1H, d, J = 7.5 Hz), 7.06–7.13 (1H, m),
7.29–7.52 (5H, m), 8.75–9.75 (1H, br), 10.60–11.50 (1H, br); MS
m/z: 337 (M+H)+.
5.1.8. 4-(2-Benzyloxyphenylmethyl)-1,2-dihydro-5-cyclopropyl-
3H-pyrazol-3-one (9h)
1H NMR (DMSO-d6) d: 3.66 (2H, s), 5.04 (2H, s), 6.87–6.92 (2H,
m), 6.98–7.07 (2H, m), 7.28–7.46 (5H, m), 10.20 (0.17H, br s), 10.74
(0.83H, br s), 12.81 (1H, s); MS m/z: 347 (MꢁH)ꢁ.
The title compound was prepared from 2-benzyloxyphenyl-
methanol and methyl 3-cyclopropyl-3-oxopropanoate, as de-
scribed for the synthesis of 9a, in 56% yield.
5.1.2. 4-(3-Benzyloxyphenylmethyl)-5-trifluoromethyl-1,2-
dihydro-3H-pyrazol-3-one (9b)
1H NMR (DMSO-d6) d: 0.55–0.64 (4H, m), 1.56–1.65 (1H, m),
3.63 (2H, s), 5.14 (2H, s), 6.82 (1H, td, J = 7.4, 1.0 Hz), 6.95–7.03
(2H, m), 7.08–7.14 (1H, m), 7.29–7.51 (5H, m), 8.50–11.60 (2H,
br); MS m/z: 321 (M+H)+.
The title compound was prepared from 3-benzyloxyphenyl-
methanol, as described for the synthesis of 9a, in 31% yield.
1H NMR (DMSO-d6) d: 3.70 (2H, s), 5.03 (2H, s), 6.66–6.77 (2H,
m), 6.80 (1H, dd, J = 8.1, 2.3 Hz), 7.16 (1H, t, J = 8.1 Hz), 7.29–7.47
(5H, m), 10.22 (0.15H, br s), 10.78 (0.85H, br s), 12.83 (1H, s); MS
m/z: 347 (MꢁH)ꢁ.
5.1.9. 4-(2-Ethylphenylmethyl)-1,2-dihydro-5-isopropyl-3H-
pyrazol-3-one (9m)
The title compound was prepared from 2-ethylphenylmethanol
and methyl 4-methyl-3-oxopentanoate, as described for the syn-
thesis of 9a, in 63% yield.
5.1.3. 4-(2-Benzyloxyphenylmethyl)-5-trifluoromethyl-1,2-
dihydro-3H-pyrazol-3-one (9c)
1H NMR (DMSO-d6) d: 1.03 (6H, d, J = 7.0 Hz), 1.14 (3H, t,
J = 7.5 Hz), 2.67 (2H, q, J = 7.5 Hz), 2.72 (1H, heptet, J = 7.0 Hz),
3.58 (2H, s), 6.92 (1H, dd, J = 7.3, 1.3 Hz), 7.00–7.15 (3H, m),
8.40–12.00 (2H, br); MS m/z: 245 (M+H)+.
The title compound was prepared from 2-benzyloxyphenyl-
methanol, as described for the synthesis of 9a, in 36% yield.
1H NMR (DMSO-d6) d: 3.76 (2H, s), 5.17 (2H, s), 6.71 (1H, d,
J = 7.1 Hz), 6.82 (1H, t, J = 7.1 Hz), 7.02 (1H, d, J = 7.8 Hz), 7.10–
7.16 (1H, m), 7.29–7.50 (5H, m), 10.11 (0.15H, br s), 10.67
(0.85H, br s), 12.87 (1H, s); MS m/z: 347 (MꢁH)ꢁ.
5.1.10. 1,2-Dihydro-4-(2-methoxyphenylmethyl)-5-isopropyl-
3H-pyrazol-3-one (9n)
For the synthesis of following aglycones (9d–h, m, n, r, t, v–y),
tetrahydrofuran (THF) was used as the solvent instead of DME.
The title compound was prepared from 2-methoxyphenylmeth-
anol and methyl 4-methyl-3-oxopentanoate, as described for the
synthesis of 9a, in 65% yield.
5.1.4. 4-(2-Benzyloxyphenylmethyl)-1,2-dihydro-5-methyl-3H-
pyrazol-3-one (9d)
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.81 (1H, heptet,
J = 7.0 Hz), 3.50 (2H, s), 3.80 (3H, s), 6.76–6.82 (1H, m), 6.87–6.93
(2H, m), 7.09–7.15 (1H, m), 8.40–12.20 (2H, br); MS m/z: 247
(M+H)+.
The title compound was prepared from 2-benzyloxyphenyl-
methanol and methyl acetoacetate, as described for the synthesis
of 9a, in 61% yield.
1H NMR (DMSO-d6) d: 1.90 (3H, s), 3.55 (2H, s), 5.14 (2H, s), 6.81
(1H, td, J = 7.4, 1.0 Hz), 6.93–7.02 (2H, m), 7.07–7.14 (1H, m), 7.29–
7.36 (1H, m), 7.37–7.43 (2H, m), 7.45–7.50 (2H, m), 8.60–11.80
(2H, br); MS m/z: 295 (M+H)+.
5.1.11. 4-(4-Ethylphenylmethyl)-1,2-dihydro-5-isopropyl-3H-
pyrazol-3-one (9r)
The title compound was prepared from 4-ethylphenylmethanol
and methyl 4-methyl-3-oxopentanoate, as described for the syn-
thesis of 9a, in 71% yield.
5.1.5. 4-(2-Benzyloxyphenylmethyl)-5-ethyl-1,2-dihydro-3H-
pyrazol-3-one (9e)
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 1.13 (3H, t,
J = 7.5 Hz), 2.53 (2H, q, J = 7.5 Hz), 2.84 (1H, heptet, J = 7.0 Hz),
3.53 (2H, s), 7.06 (4H, s), 8.40–12.00 (2H, br); MS m/z: 245 (M+H)+.
The title compound was prepared from 2-benzyloxyphenyl-
methanol and methyl 3-oxopentanoate, as described for the syn-
thesis of 9a, in 74% yield.
5.1.12. 4-(4-Benzyloxyphenylmethyl)-1,2-dihydro-5-isopropyl-
3H-pyrazol-3-one (9t)
1H NMR (DMSO-d6) d: 0.94 (3H, t, J = 7.6 Hz), 2.31 (2H, q,
J = 7.6 Hz), 3.56 (2H, s), 5.14 (2H, s), 6.81 (1H, td, J = 7.5, 0.9 Hz),
6.95 (1H, dd, J = 7.5, 1.5 Hz), 7.00 (1H, dd, J = 8.2, 0.9 Hz), 7.07–
7.14 (1H, m), 7.29–7.50 (5H, m), 8.60–11.90 (2H, br); MS m/z:
309 (M+H)+.
The title compound was prepared from 4-benzyloxyphenyl-
methanol and methyl 4-methyl-3-oxopentanoate, as described
for the synthesis of 9a, in 70% yield.
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.83 (1H, heptet,
J = 7.0 Hz), 3.51 (2H, s), 5.03 (2H, s), 6.85–6.90 (2H, m), 7.03–7.08
(2H, m), 7.28–7.45 (5H, m), 8.40–12.20 (2H, br); MS m/z: 323
(M+H)+.
5.1.6. 4-(2-Benzyloxyphenylmethyl)-1,2-dihydro-5-isopropyl-
3H-pyrazol-3-one (9f)
The title compound was prepared from 2-benzyloxyphenyl-
methanol and methyl 4-methyl-3-oxopentanoate, as described
for the synthesis of 9a, in 67% yield.
5.1.13. 1,2-Dihydro-4-(3-methylphenylmethyl)-5-isopropyl-3H-
pyrazol-3-one (9v)
1H NMR (DMSO-d6) d: 1.11 (6H, d, J = 7.0 Hz), 2.77 (1H, heptet,
J = 7.0 Hz), 3.58 (2H, s), 5.14 (2H, s), 6.81 (1H, td, J = 7.4, 1.0 Hz),
6.94 (1H, dd, J = 7.4, 1.5 Hz), 7.00 (1H, dd, J = 8.2, 1.0 Hz), 7.07–
7.14 (1H, m), 7.30–7.51 (5H, m), 8.50–12.00 (2H, br); MS m/z:
323 (M+H)+.
The title compound was prepared from 3-methylphenylmetha-
nol and methyl 4-methyl-3-oxopentanoate, as described for the
synthesis of 9a, in 22% yield.
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.23 (3H, s), 2.82
(1H, heptet, J = 7.0 Hz), 3.53 (2H, s), 6.90–6.97 (3H, m), 7.11 (1H, t,

N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
6605
J = 7.4 Hz), 8.90–10.00 (1H, br), 10.55–11.50 (1H, br); MS m/z: 231
1H NMR (DMSO-d6) d: 1.04 (6H, d, J = 7.0 Hz), 2.28 (3H, s), 2.74
(1H, heptet, J = 7.0 Hz), 3.52 (2H, s), 6.87–6.93 (1H, m), 7.00–7.13
(3H, m), 8.40–12.00 (2H, br); MS m/z: 231 (M+H)+.
(M+H)+.
5.1.14. 4-(3-Benzyloxyphenylmethyl)-1,2-dihydro-5-isopropyl-
3H-pyrazol-3-one (9w)
5.1.20. 4-(2-Trifluoromethylphenylmethyl)-1,2-dihydro-5-
isopropyl-3H-pyrazol-3-one (9l)
The title compound was prepared from 3-benzyloxyphenyl-
methanol and methyl 4-methyl-3-oxopentanoate, as described
for the synthesis of 9a, in 75% yield.
The title compound was prepared from 2-trifluoromethylphe-
nylmethyl bromide, as described for the synthesis of 9i, in 28%
yield.
1H NMR (DMSO-d6) d: 1.06 (6H, d, J = 7.0 Hz), 2.82 (1H, heptet,
J = 7.0 Hz), 3.54 (2H, s), 5.03 (2H, s), 6.71–6.81 (3H, m), 7.14 (1H,
t, J = 7.8 Hz), 7.28–7.44 (5H, m), 8.80–10.20 (1H, br), 10.30–11.70
(1H, br); MS m/z: 323 (M+H)+.
1H NMR (DMSO-d6) d: 1.05 (6H, d, J = 7.0 Hz), 2.71 (1H, heptet,
J = 7.0 Hz), 3.78 (2H, s), 7.13 (1H, d, J = 7.8 Hz), 7.37 (1H, t,
J = 7.8 Hz), 7.54 (1H, t, J = 7.8 Hz), 7.68 (1H, d, J = 7.8 Hz), 8.90–
12.10 (2H, br); MS m/z: 285 (M+H)+.
5.1.15. 1,2-Dihydro-4-(2,4-dimethylphenylmethyl)-5-isopropyl-
3H-pyrazol-3-one (9x)
5.1.21. 4-(2-Cyanophenylmethyl)-1,2-dihydro-5-isopropyl-3H-
pyrazol-3-one (9o)
The title compound was prepared from 2,4-dimethylphenyl-
methanol and methyl 4-methyl-3-oxopentanoate, as described
for the synthesis of 9a, in 74% yield.
The title compound was prepared from 2-cyanophenylmethyl
bromide, as described for the synthesis of 9i, in 43% yield. In the
cyclization step, EtOH was used as the solvent, and the reaction
was carried out at room temperature.
1H NMR (DMSO-d6) d: 1.04 (6H, d, J = 7.0 Hz), 2.20 (3H, s), 2.24
(3H, s), 2.72 (1H, heptet, J = 7.0 Hz), 3.46 (2H, s), 6.77 (1H, d,
J = 7.8 Hz), 6.84 (1H, d, J = 7.8 Hz), 6.91 (1H, s), 8.60–10.10 (1H,
br), 10.40–11.90 (1H, br); MS m/z: 245 (M+H)+.
1H NMR (DMSO-d6) d: 1.09 (6H, d, J = 7.0 Hz), 2.87 (1H, heptet,
J = 7.0 Hz), 3.79 (2H, s), 7.24 (1H, d, J = 7.7 Hz), 7.33–7.39 (1H, m),
7.59 (1H, td, J = 7.7, 1.3 Hz), 7.76 (1H, dd, J = 7.7, 1.3 Hz), 8.40–
12.20 (2H, br); MS m/z: 242 (M+H)+.
5.1.16. 4-(4-Benzyloxy-2-methylphenylmethyl)-1,2-dihydro-5-
isopropyl-3H-pyrazol-3-one (9y)
The title compound was prepared from 4-benzyloxy-2-methyl-
phenylmethanol and methyl 4-methyl-3-oxopentanoate, as de-
scribed for the synthesis of 9a, in 80% yield.
5.1.22. 4-(4-Fluorophenylmethyl)-1,2-dihydro-5-isopropyl-3H-
pyrazol-3-one (9p)
The title compound was prepared from 4-fluorophenylmethyl
bromide, as described for the synthesis of 9i, in 58% yield.
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.83 (1H, heptet,
J = 7.0 Hz), 3.56 (2H, s), 7.01–7.09 (2H, m), 7.13–7.20 (2H, m),
8.40–12.20 (2H, br); MS m/z: 235 (M+H)+.
1H NMR (DMSO-d6) d: 1.04 (6H, d, J = 7.0 Hz), 2.24 (3H, s), 2.73
(1H, heptet, J = 7.0 Hz), 3.44 (2H, s), 5.02 (2H, s), 6.70 (1H, dd,
J = 8.4, 2.6 Hz), 6.76–6.81 (2H, m), 7.27–7.45 (5H, m), 8.70–9.90
(1H, br), 10.70–11.60 (1H, br); MS m/z: 337 (M+H)+.
5.1.17. 4-Benzyl-1,2-dihydro-5-isopropyl-3H-pyrazol-3-one (9i)
NaH (916 mg, 22.9 mmol, 60% oil dispersion), under ice cooling,
was added to a solution of methyl 4-methyl-3-oxopentanoate
(3.00 g, 20.8 mmol) in THF (25 mL), and the mixture was stirred
for 5 min. Benzyl bromide (2.48 mL, 20.8 mmol) was added, and
the mixture was refluxed for 5 h. After cooling to room tempera-
ture, 1 M HCl (25 mL) was added, and the resulting mixture was ex-
tracted with EtOAc. The extract was washed with brine and dried
over anhydrous MgSO4. The solvent was removed under reduced
pressure to give crude methyl 2-benzyl-4-methyl-3-oxopentanoate
(8i). This material was dissolved in toluene (20 mL), and N2H4ꢀH2O
(2.02 mL, 41.6 mmol) was added. After refluxing for 10 h, water was
added, and the mixture was extracted with EtOAc. The extract was
washed with brine and dried over anhydrous MgSO4. The solvent
was removed under reduced pressure, and the residue was purified
by column chromatography on silica gel (eluent: CH2Cl2/
MeOH = 10:1) to give 9i (3.43 g, 76%) as a white solid.
5.1.23. 1,2-Dihydro-4-(4-methylphenylmethyl)-5-isopropyl-3H-
pyrazol-3-one (9q)
The title compound was prepared from 4-methylphenylmethyl
bromide, as described for the synthesis of 9i, in 33% yield.
1H NMR (DMSO-d6) d: 1.06 (6H, d, J = 7.0 Hz), 2.23 (3H, s), 2.82
(1H, heptet, J = 7.0 Hz), 3.52 (2H, s), 7.03 (4H, s), 8.40–12.20 (2H,
br); MS m/z: 231 (M+H)+.
5.1.24. 1,2-Dihydro-4-(4-methoxyphenylmethyl)-5-isopropyl-
3H-pyrazol-3-one (9s)
The title compound was prepared from 4-methoxyphenylmeth-
yl chloride, as described for the synthesis of 9i, in 61% yield.
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.83 (1H, heptet,
J = 7.0 Hz), 3.51 (2H, s), 3.69 (3H, s), 6.77–6.82 (2H, m), 7.03–7.08
(2H, m), 8.40–12.20 (2H, br); MS m/z: 247 (M+H)+.
5.1.25. 4-(4-Cyanophenylmethyl)-1,2-dihydro-5-isopropyl-3H-
pyrazol-3-one (9u)
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.84 (1H, heptet,
J = 7.0 Hz), 3.58 (2H, s), 7.08–7.18 (3H, m), 7.19–7.26 (2H, m),
8.40–12.00 (2H, br); MS m/z: 217 (M+H)+.
The title compound was prepared from 4-cyanophenylmethyl
bromide, as described for the synthesis of 9i, in 33% yield. In the
cyclization step, EtOH was used as the solvent, and the reaction
was carried out at room temperature.
5.1.18. 4-(2-Fluorophenylmethyl)-1,2-dihydro-5-isopropyl-3H-
pyrazol-3-one (9j)
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.84 (1H, heptet,
J = 7.0 Hz), 3.67 (2H, s), 7.31–7.37 (2H, m), 7.69–7.74 (2H, m),
8.80–10.30 (1H, br), 10.40–11.90 (1H, br); MS m/z: 242 (M+H)+.
The title compound was prepared from 2-fluorophenylmethyl
bromide, as described for the synthesis of 9i, in 71% yield.
1H NMR (DMSO-d6) d: 1.08 (6H, d, J = 7.0 Hz), 2.83 (1H, heptet,
J = 7.0 Hz), 3.58 (2H, s), 7.04–7.23 (4H, m), 8.90–10.10 (1H, br),
10.50–11.70 (1H, br); MS m/z: 235 (M+H)+.
5.1.26. 4-(4-Benzyloxyphenylmethyl)-5-trifluoromethyl-1H-
pyrazol-3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10a)
Aqueous 5 M NaOH (3.80 mL, 18.8 mmol) was added to a mix-
ture of 9a (1.64 g, 4.71 mmol), acetobromoglucose (1.94 g,
4.71 mmol), and BnN(n-Bu)3Cl (1.47 g, 4.71 mmol) in CH2Cl2
(30 mL), and the mixture was stirred at room temperature over-
night. 1 M HCl (25 mL) was added, and the resulting mixture was
5.1.19. 1,2-Dihydro-4-(2-methylphenylmethyl)-5-isopropyl-3H-
pyrazol-3-one (9k)
The title compound was prepared from 2-methylphenylmethyl
bromide, as described for the synthesis of 9i, in 45% yield.

6606
N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
extracted with EtOAc. The extract was washed with brine and
dried over anhydrous MgSO4. The solvent was removed under re-
duced pressure, and the residue was purified by column chroma-
tography on silica gel (eluent: hexane/EtOAc = 2:1–1:1) to give
10a (266 mg, 8%) as a white solid.
(1H, d, J = 7.5 Hz), 7.07–7.14 (1H, m), 7.30–7.50 (5H, m), 11.70
(1H, s); MS m/z: 653 (M+H)+.
5.1.32. 4-(2-Benzyloxyphenylmethyl)-5-tert-butyl-1H-pyrazol-
3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10g)
1H NMR (CDCl3) d: 1.89 (3H, s), 2.02 (3H, s), 2.04 (3H, s), 2.07
(3H, s), 3.78 (2H, s), 3.79–3.85 (1H, m), 4.20 (1H, dd, J = 12.5,
2.0 Hz), 4.26 (1H, dd, J = 12.5, 4.3 Hz), 5.02 (2H, s), 5.16–5.30 (3H,
m), 5.38 (1H, br s), 6.83–6.89 (2H, m), 7.04–7.10 (2H, m), 7.27–
7.44 (5H, m), 9.00–11.00 (1H, br); MS m/z: 677 (MꢁH)ꢁ.
The title compound was prepared from 9g, as described for the
synthesis of 10a, in 28% yield.
1H NMR (CDCl3) d: 1.22 (9H, s), 1.70 (3H, s), 1.97 (3H, s), 2.01
(3H, s), 2.04 (3H, s), 3.77–3.92 (3H, m), 4.12 (1H, dd, J = 12.5,
2.1 Hz), 4.29 (1H, dd, J = 12.5, 3.8 Hz), 5.11–5.26 (5H, m), 5.51
(1H, d, J = 8.0 Hz), 6.75–6.82 (2H, m), 6.88 (1H, d, J = 8.0 Hz),
7.06–7.13 (1H, m), 7.29–7.50 (5H, m), 8.30–9.90 (1H, br); MS m/
z: 667 (M+H)+.
5.1.27. 4-(3-Benzyloxyphenylmethyl)-5-trifluoromethyl-1H-
pyrazol-3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10b)
The title compound was prepared from 9b, as described for the
synthesis of 10a, in 9% yield.
5.1.33. 4-(2-Benzyloxyphenylmethyl)-5-cyclopropyl-1H-
1H NMR (CDCl3) d: 1.87 (3H, s), 2.02 (3H, s), 2.04 (3H, s), 2.06
(3H, s), 3.75 (2H, s), 3.77–3.84 (1H, m), 4.15–4.28 (2H, m), 5.02
(2H, s), 5.12–5.48 (4H, m), 6.73–6.82 (3H, m), 7.16 (1H, t,
J = 8.0 Hz), 7.27–7.44 (5H, m), 9.30–11.00 (1H, br); MS m/z: 677
(MꢁH)ꢁ.
pyrazol-3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10h)
The title compound was prepared from 9h, as described for the
synthesis of 10a, in 29% yield.
1H NMR (DMSO-d6) d: 0.53–0.65 (4H, m), 1.57–1.68 (1H, m),
1.80 (3H, s), 1.93 (3H, s), 1.97 (3H, s), 1.98 (3H, s), 3.60 (2H, s),
3.93 (1H, dd, J = 12.3, 2.3 Hz), 4.04–4.11 (1H, m), 4.16 (1H, dd,
J = 12.3, 4.5 Hz), 4.91–4.99 (2H, m), 5.13 (2H, s), 5.39 (1H, t,
J = 9.7 Hz), 5.67 (1H, d, J = 8.0 Hz), 6.76–6.83 (1H, m), 6.90 (1H,
dd, J = 7.5, 1.8 Hz), 7.00 (1H, d, J = 7.3 Hz), 7.08–7.15 (1H, m),
7.29–7.49 (5H, m), 11.49 (1H, s); MS m/z: 651 (M+H)+.
5.1.28. 4-(2-Benzyloxyphenylmethyl)-5-trifluoromethyl-1H-
pyrazol-3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10c)
The title compound was prepared from 9c, as described for the
synthesis of 10a, in 7% yield.
1H NMR (CDCl3) d: 1.76 (3H, s), 1.99 (3H, s), 2.03 (3H, s), 2.07
(3H, s), 3.69–3.77 (1H, m), 3.82 (1H, d, J = 16.8 Hz), 3.90 (1H, d,
J = 16.8 Hz), 4.16 (1H, dd, J = 12.4, 1.9 Hz), 4.23 (1H, dd, J = 12.4,
4.4 Hz), 5.08–5.33 (6H, m), 6.80–6.90 (3H, m), 7.11–7.17 (1H, m),
7.29–7.41 (5H, m), 9.40–10.70 (1H, br); MS m/z: 677 (MꢁH)ꢁ.
5.1.34. 4-Benzyl-5-isopropyl-1H-pyrazol-3-yl 2,3,4,6-tetra-O-
acetyl-b-D-glucopyranoside (10i)
The title compound was prepared from 9i, as described for the
synthesis of 10a, in 26% yield.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 6.9 Hz), 1.08 (3H, d,
J = 6.9 Hz), 1.89 (3H, s), 1.95 (3H, s), 1.985 (3H, s), 1.988 (3H, s),
2.89 (1H, heptet, J = 6.9 Hz), 3.53 (1H, d, J = 15.8 Hz), 3.58 (1H, d,
J = 15.8 Hz), 3.97 (1H, dd, J = 12.2, 2.1 Hz), 4.09–4.16 (1H, m),
4.19 (1H, dd, J = 12.2, 4.6 Hz), 4.95–5.03 (2H, m), 5.42 (1H, t,
J = 9.5 Hz), 5.73 (1H, d, J = 8.0 Hz), 7.07–7.25 (5H, m), 11.72 (1H,
s); MS m/z: 547 (M+H)+.
5.1.29. 4-(2-Benzyloxyphenylmethyl)-5-methyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10d)
The title compound was prepared from 9d, as described for the
synthesis of 10a, in 23% yield.
1H NMR (DMSO-d6) d: 1.80 (3H, s), 1.92 (3H, s), 1.94 (3H, s), 1.97
(3H, s), 1.98 (3H, s), 3.52 (2H, s), 3.93 (1H, dd, J = 12.2, 2.1 Hz),
4.05–4.12 (1H, m), 4.17 (1H, dd, J = 12.2, 4.5 Hz), 4.92–4.99 (2H,
m), 5.13 (2H, s), 5.39 (1H, t, J = 9.5 Hz), 5.67 (1H, d, J = 8.3 Hz),
6.76–6.82 (1H, m), 6.89 (1H, dd, J = 7.4, 1.6 Hz), 6.99 (1H, d,
J = 7.3 Hz), 7.08–7.14 (1H, m), 7.29–7.48 (5H, m), 11.66 (1H, s);
MS m/z: 625 (M+H)+.
5.1.35. 4-(2-Fluorophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10j)
The title compound was prepared from 9j, as described for the
synthesis of 10a, in 20% yield.
1H NMR (DMSO-d6) d: 1.08 (3H, d, J = 7.3 Hz), 1.10 (3H, d,
J = 7.3 Hz), 1.87 (3H, s), 1.94 (3H, s), 1.98 (6H, s), 2.88 (1H, heptet,
J = 7.3 Hz), 3.54 (1H, d, J = 16.1 Hz), 3.59 (1H, d, J = 16.1 Hz), 3.96
(1H, dd, J = 12.1, 2.0 Hz), 4.08–4.15 (1H, m), 4.18 (1H, dd, J = 12.1,
4.6 Hz), 4.93–5.01 (2H, m), 5.41 (1H, t, J = 9.5 Hz), 5.72 (1H, d,
J = 8.3 Hz), 7.00–7.14 (3H, m), 7.15–7.24 (1H, m), 11.78 (1H, s);
MS m/z: 565 (M+H)+.
5.1.30. 4-(2-Benzyloxyphenylmethyl)-5-ethyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10e)
The title compound was prepared from 9e, as described for the
synthesis of 10a, in 27% yield.
1H NMR (DMSO-d6) d: 0.92 (3H, t, J = 7.5 Hz), 1.81 (3H, s), 1.94
(3H, s), 1.97 (3H, s), 1.98 (3H, s), 2.32 (2H, q, J = 7.5 Hz), 3.53 (2H,
s), 3.94 (1H, dd, J = 12.3, 2.1 Hz), 4.05–4.12 (1H, m), 4.17 (1H, dd,
J = 12.3, 4.6 Hz), 4.92–5.00 (2H, m), 5.13 (2H, s), 5.39 (1H, t,
J = 9.5 Hz), 5.69 (1H, d, J = 8.0 Hz), 6.78 (1H, t, J = 7.2 Hz), 6.89
(1H, dd, J = 7.2, 1.6 Hz), 6.99 (1H, d, J = 7.8 Hz), 7.08–7.14 (1H, m),
7.30–7.49 (5H, m), 11.70 (1H, s); MS m/z: 639 (M+H)+.
5.1.36. 4-(2-Methylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10k)
The title compound was prepared from 9k, as described for the
synthesis of 10a, in 30% yield.
1H NMR (DMSO-d6) d: 1.03 (3H, d, J = 7.0 Hz), 1.07 (3H, d,
J = 7.0 Hz), 1.81 (3H, s), 1.93 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.24 (3H, s), 2.77 (1H, heptet, J = 7.0 Hz), 3.48 (1H, d, J = 16.8 Hz),
3.54 (1H, d, J = 16.8 Hz), 3.94 (1H, dd, J = 12.3, 2.1 Hz), 4.06–4.13
(1H, m), 4.16 (1H, dd, J = 12.3, 4.6 Hz), 4.89–4.98 (2H, m), 5.39
(1H, t, J = 9.7 Hz), 5.69 (1H, d, J = 8.3 Hz), 6.80–6.87 (1H, m),
6.99–7.13 (3H, m), 11.76 (1H, s); MS m/z: 561 (M+H)+.
5.1.31. 4-(2-Benzyloxyphenylmethyl)-5-isopropyl-1H-pyrazol-
3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10f)
The title compound was prepared from 9f, as described for the
synthesis of 10a, in 32% yield.
1H NMR (DMSO-d6) d: 0.988 (3H, d, J = 7.0 Hz), 0.992 (3H, d,
J = 7.0 Hz), 1.81 (3H, s), 1.94 (3H, s), 1.97 (3H, s), 1.98 (3H, s),
2.80 (1H, heptet, J = 7.0 Hz), 3.55 (2H, s), 3.94 (1H, dd, J = 12.3,
2.1 Hz), 4.06–4.13 (1H, m), 4.17 (1H, dd, J = 12.3, 4.6 Hz), 4.92–
4.99 (2H, m), 5.13 (2H, s), 5.40 (1H, t, J = 9.7 Hz), 5.70 (1H, d,
J = 8.3 Hz), 6.75–6.81 (1H, m), 6.86 (1H, dd, J = 7.5, 1.5 Hz), 7.00
5.1.37. 4-(2-Trifluoromethylphenylmethyl)-5-isopropyl-1H-
pyrazol-3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10l)
The title compound was prepared from 9l, as described for the
synthesis of 10a, in 28% yield.

N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
6607
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 1.79 (3H, s), 1.92 (3H, s), 1.98 (6H, s), 2.73 (1H, heptet,
J = 7.0 Hz), 3.73 (1H, d, J = 17.1 Hz), 3.79 (1H, d, J = 17.1 Hz), 3.92
(1H, dd, J = 11.8, 1.5 Hz), 4.07–4.18 (2H, m), 4.86–4.97 (2H, m),
5.39 (1H, t, J = 9.5 Hz), 5.69 (1H, d, J = 8.0 Hz), 7.02 (1H, d,
J = 7.7 Hz), 7.38 (1H, t, J = 7.7 Hz), 7.51 (1H, t, J = 7.7 Hz), 7.68
(1H, d, J = 7.7 Hz), 11.91 (1H, s); MS m/z: 615 (M+H)+.
(1H, m), 4.19 (1H, dd, J = 12.2, 4.6 Hz), 4.94–5.02 (2H, m), 5.42
(1H, t, J = 9.7 Hz), 5.73 (1H, d, J = 8.3 Hz), 6.97 (2H, d, J = 7.9 Hz),
7.06 (2H, d, J = 7.9 Hz), 11.71 (1H, s); MS m/z: 561 (M+H)+.
5.1.43. 4-(4-Ethylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10r)
The title compound was prepared from 9r, as described for the
synthesis of 10a, in 14% yield.
5.1.38. 4-(2-Ethylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10m)
The title compound was prepared from 9m, as described for the
synthesis of 10a, in 29% yield.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 6.9 Hz), 1.09 (3H, d,
J = 6.9 Hz), 1.12 (3H, t, J = 7.5 Hz), 1.88 (3H, s), 1.95 (3H, s), 1.985
(3H, s), 1.987 (3H, s), 2.52 (2H, q, J = 7.5 Hz), 2.89 (1H, heptet,
J = 6.9 Hz), 3.48 (1H, d, J = 15.8 Hz), 3.53 (1H, d, J = 15.8 Hz), 3.97
(1H, dd, J = 12.2, 2.3 Hz), 4.08–4.15 (1H, m), 4.19 (1H, dd, J = 12.2,
4.6 Hz), 4.94–5.03 (2H, m), 5.42 (1H, t, J = 9.7 Hz), 5.72 (1H, d,
J = 8.3 Hz), 7.00 (2H, d, J = 7.5 Hz), 7.04 (2H, d, J = 7.5 Hz), 11.70
(1H, s); MS m/z: 575 (M+H)+.
1H NMR (DMSO-d6) d: 1.03 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 1.12 (3H, t, J = 7.5 Hz), 1.80 (3H, s), 1.93 (3H, s), 1.98
(3H, s), 1.99 (3H, s), 2.63 (2H, q, J = 7.5 Hz), 2.75 (1H, heptet,
J = 7.0 Hz), 3.53 (1H, d, J = 16.6 Hz), 3.60 (1H, d, J = 16.6 Hz), 3.93
(1H, dd, J = 12.1, 2.0 Hz), 4.06–4.13 (1H, m), 4.17 (1H, dd, J = 12.1,
4.6 Hz), 4.89–4.99 (2H, m), 5.40 (1H, t, J = 9.7 Hz), 5.69 (1H, d,
J = 8.3 Hz), 6.83 (1H, d, J = 7.3 Hz), 7.02 (1H, td, J = 7.3, 1.7 Hz),
7.05–7.15 (2H, m), 11.77 (1H, s); MS m/z: 575 (M+H)+.
5.1.44. 4-(4-Methoxyphenylmethyl)-5-isopropyl-1H-pyrazol-3-
yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10s)
The title compound was prepared from 9s, as described for the
synthesis of 10a, in 27% yield.
5.1.39. 4-(2-Methoxyphenylmethyl)-5-isopropyl-1H-pyrazol-3-
yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10n)
The title compound was prepared from 9n, as described for the
synthesis of 10a, in 30% yield.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 7.0 Hz), 1.08 (3H, d,
J = 7.0 Hz), 1.90 (3H, s), 1.95 (3H, s), 1.986 (3H, s), 1.988 (3H, s),
2.88 (1H, heptet, J = 7.0 Hz), 3.46 (1H, d, J = 15.8 Hz), 3.50 (1H, d,
J = 15.8 Hz), 3.69 (3H, s), 3.97 (1H, dd, J = 12.3, 2.0 Hz), 4.08–4.16
(1H, m), 4.19 (1H, dd, J = 12.3, 4.6 Hz), 4.94–5.03 (2H, m), 5.42
(1H, t, J = 9.5 Hz), 5.72 (1H, d, J = 8.3 Hz), 6.77 (2H, d, J = 8.7 Hz),
7.01 (2H, d, J = 8.7 Hz), 11.69 (1H, s); MS m/z: 577 (M+H)+.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 7.0 Hz), 1.09 (3H, d,
J = 7.0 Hz), 1.83 (3H, s), 1.94 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.87 (1H, heptet, J = 7.0 Hz), 3.48 (2H, s), 3.79 (3H, s), 3.94 (1H,
dd, J = 12.3, 2.1 Hz), 4.05–4.13 (1H, m), 4.17 (1H, dd, J = 12.3,
4.6 Hz), 4.92–4.99 (2H, m), 5.39 (1H, t, J = 9.7 Hz), 5.69 (1H, d,
J = 8.3 Hz), 6.74–6.85 (2H, m), 6.91 (1H, d, J = 8.3 Hz), 7.09–7.16
(1H, m), 11.72 (1H, s); MS m/z: 577 (M+H)+.
5.1.45. 4-(4-Benzyloxyphenylmethyl)-5-isopropyl-1H-pyrazol-
3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10t)
The title compound was prepared from 9t, as described for the
synthesis of 10a, in 36% yield.
5.1.40. 4-(2-Cyanophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10o)
The title compound was prepared from 9o, as described for the
synthesis of 10a, in 30% yield.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 7.0 Hz), 1.08 (3H, d,
J = 7.0 Hz), 1.88 (3H, s), 1.95 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.88 (1H, heptet, J = 7.0 Hz), 3.46 (1H, d, J = 15.9 Hz), 3.50 (1H, d,
J = 15.9 Hz), 3.97 (1H, dd, J = 12.2, 1.9 Hz), 4.09–4.16 (1H, m),
4.19 (1H, dd, J = 12.2, 4.5 Hz), 4.94–5.02 (2H, m), 5.03 (2H, s),
5.42 (1H, t, J = 9.7 Hz), 5.72 (1H, d, J = 8.0 Hz), 6.82–6.88 (2H, m),
6.98–7.03 (2H, m), 7.27–7.44 (5H, m), 11.70 (1H, s); MS m/z: 653
(M+H)+.
1H NMR (DMSO-d6) d: 1.09 (3H, d, J = 7.0 Hz), 1.10 (3H, d,
J = 7.0 Hz), 1.85 (3H, s), 1.94 (3H, s), 1.98 (6H, s), 2.91 (1H, heptet,
J = 7.0 Hz), 3.75 (1H, d, J = 16.3 Hz), 3.80 (1H, d, J = 16.3 Hz), 3.94
(1H, dd, J = 12.1, 1.8 Hz), 4.08–4.15 (1H, m), 4.16 (1H, dd, J = 12.1,
4.6 Hz), 4.91–4.99 (2H, m), 5.40 (1H, t, J = 9.7 Hz), 5.72 (1H, d,
J = 8.0 Hz), 7.16 (1H, d, J = 7.7 Hz), 7.34–7.40 (1H, m), 7.53–7.59
(1H, m), 7.76 (1H, dd, J = 7.7, 1.3 Hz), 11.88 (1H, s); MS m/z: 572
(M+H)+.
5.1.46. 4-(4-Cyanophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10u)
The title compound was prepared from 9u, as described for the
synthesis of 10a, in 32% yield.
5.1.41. 4-(4-Fluorophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10p)
The title compound was prepared from 9p, as described for the
synthesis of 10a, in 39% yield.
1H NMR (DMSO-d6) d: 1.079 (3H, d, J = 7.0 Hz), 1.082 (3H, d,
J = 7.0 Hz), 1.89 (3H, s), 1.94 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.90 (1H, heptet, J = 7.0 Hz), 3.64 (1H, d, J = 16.9 Hz), 3.68 (1H, d,
J = 16.9 Hz), 3.96 (1H, dd, J = 12.0, 1.9 Hz), 4.09–4.16 (1H, m), 4.18
(1H, dd, J = 12.0, 4.7 Hz), 4.93–5.00 (2H, m), 5.42 (1H, t,
J = 9.5 Hz), 5.73 (1H, d, J = 8.0 Hz), 7.29 (2H, d, J = 8.3 Hz), 7.69
(2H, d, J = 8.3 Hz), 11.82 (1H, s); MS m/z: 572 (M+H)+.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 7.0 Hz), 1.08 (3H, d,
J = 7.0 Hz), 1.90 (3H, s), 1.95 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.89 (1H, heptet, J = 7.0 Hz), 3.52 (1H, d, J = 16.6 Hz), 3.56 (1H, d,
J = 16.6 Hz), 3.97 (1H, dd, J = 12.1, 2.0 Hz), 4.09–4.16 (1H, m), 4.19
(1H, dd, J = 12.1, 4.5 Hz), 4.94–5.02 (2H, m), 5.42 (1H, t,
J = 9.7 Hz), 5.73 (1H, d, J = 8.3 Hz), 6.99–7.15 (4H, m), 11.74 (1H,
s); MS m/z: 565 (M+H)+.
5.1.47. 4-(3-Methylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10v)
The title compound was prepared from 9v, as described for the
synthesis of 10a, in 34% yield.
5.1.42. 4-(4-Methylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside (10q)
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 7.0 Hz), 1.08 (3H, d,
J = 7.0 Hz), 1.88 (3H, s), 1.95 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.23 (3H, s), 2.88 (1H, heptet, J = 7.0 Hz), 3.49 (1H, d, J = 15.8 Hz),
3.53 (1H, d, J = 15.8 Hz), 3.96 (1H, dd, J = 12.2, 2.1 Hz), 4.09–4.16
(1H, m), 4.19 (1H, dd, J = 12.2, 4.5 Hz), 4.94–5.03 (2H, m), 5.42
(1H, t, J = 9.5 Hz), 5.72 (1H, d, J = 8.3 Hz), 6.87 (1H, d, J = 7.8 Hz),
6.91 (1H, s), 6.93 (1H, d, J = 7.8 Hz), 7.09 (1H, t, J = 7.8 Hz), 11.71
(1H, s); MS m/z: 561 (M+H)+.
The title compound was prepared from 9q, as described for the
synthesis of 10a, in 28% yield.
1H NMR (DMSO-d6) d: 1.06 (3H, d, J = 6.9 Hz), 1.08 (3H, d,
J = 6.9 Hz), 1.89 (3H, s), 1.95 (3H, s), 1.987 (3H, s), 1.988 (3H, s),
2.22 (3H, s), 2.87 (1H, heptet, J = 6.9 Hz), 3.47 (1H, d, J = 15.9 Hz),
3.52 (1H, d, J = 15.9 Hz), 3.97 (1H, dd, J = 12.2, 2.1 Hz), 4.09–4.16

6608
N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
5.1.48. 4-(3-Benzyloxyphenylmethyl)-5-isopropyl-1H-pyrazol-
3-yl 2,3,4,6-tetra-O-acetyl-b- -glucopyranoside (10w)
5.1.53. 4-(2-Benzyloxyphenylmethyl)-5-trifluoromethyl-1H-
pyrazol-3-yl b- -glucopyranoside (11c)
D
D
The title compound was prepared from 9w, as described for the
synthesis of 10a, in 31% yield.
The title compound was prepared from 10c, as described for the
synthesis of 11a, in 55% yield.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.07 (3H, d,
J = 7.0 Hz), 1.86 (3H, s), 1.94 (3H, s), 1.96 (3H, s), 1.99 (3H, s),
2.86 (1H, heptet, J = 7.0 Hz), 3.49 (1H, d, J = 15.9 Hz), 3.54 (1H, d,
J = 15.9 Hz), 3.96 (1H, dd, J = 12.2, 2.1 Hz), 4.09–4.16 (1H, m),
4.19 (1H, dd, J = 12.2, 4.5 Hz), 4.94–5.03 (2H, m), 5.03 (2H, s),
5.42 (1H, t, J = 9.7 Hz), 5.73 (1H, d, J = 8.0 Hz), 6.68 (1H, d,
J = 7.8 Hz), 6.73 (1H, d, J = 2.1 Hz), 6.78 (1H, dd, J = 7.8, 2.1 Hz),
7.13 (1H, t, J = 7.8 Hz), 7.28–7.44 (5H, m), 11.73 (1H, s); MS m/z:
653 (M+H)+.
1H NMR (DMSO-d6) d: 3.06–3.20 (4H, m), 3.46 (1H, d,
J = 11.2 Hz), 3.64 (1H, d, J = 11.2 Hz), 3.82 (2H, s), 4.50 (1H, br s),
4.60–4.86 (1H, br), 5.00 (1H, br s), 5.07 (1H, br s), 5.12–5.54 (3H,
m), 6.75–6.85 (2H, m), 7.01 (1H, d, J = 8.0 Hz), 7.09–7.16 (1H, m),
7.29–7.35 (1H, m), 7.36–7.49 (4H, m), 13.34 (1H, br s); MS m/z:
511 (M+H)+. HRMS (FAB+) calcd for C24H26F3N2O7 511.1687; found
511.1686 (M+H)+.
5.1.54. 4-(2-Benzyloxyphenylmethyl)-5-methyl-1H-pyrazol-3-yl
b-D
-glucopyranoside (11d)
5.1.49. 4-(2,4-Dimethylphenylmethyl)-5-isopropyl-1H-pyrazol-
The title compound was prepared from 10d, as described for the
3-yl 2,3,4,6-tetra-O-acetyl-b-
The title compound was prepared from 9x, as described for the
synthesis of 10a, in 24% yield.
D
-glucopyranoside (10x)
synthesis of 11a, in 64% yield.
1H NMR (DMSO-d6) d: 1.92 (3H, s), 3.05–3.23 (4H, m), 3.40–3.49
(1H, m), 3.54–3.66 (3H, m), 4.42 (1H, t, J = 5.8 Hz), 4.91 (1H, d,
J = 4.8 Hz), 4.98 (1H, d, J = 4.5 Hz), 5.10 (1H, d, J = 5.0 Hz), 5.15
(2H, s), 5.19 (1H, d, J = 7.5 Hz), 6.82 (1H, td, J = 7.5, 0.8 Hz), 6.97–
7.03 (2H, m), 7.07–7.14 (1H, m), 7.29–7.36 (1H, m), 7.37–7.43
(2H, m), 7.45–7.50 (2H, m), 11.50 (1H, s); 13C NMR (100 MHz,
CD3OD) d: 10.1, 22.6, 62.7, 71.1, 71.3, 75.0, 78.0, 78.4, 102.5,
103.3, 112.8, 121.7, 128.1, 128.6, 128.9, 129.6, 130.6, 130.8,
139.0, 140.0, 157.7, 162.0; MS m/z: 457 (M+H)+; HRMS (FAB+)
calcd for C24H29N2O7 457.1969; found 457.1969 (M+H)+.
1H NMR (DMSO-d6) d: 1.04 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 1.82 (3H, s), 1.93 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.193 (3H, s), 2.199 (3H, s), 2.76 (1H, heptet, J = 7.0 Hz), 3.42 (1H,
d, J = 16.6 Hz), 3.48 (1H, d, J = 16.6 Hz), 3.94 (1H, dd, J = 12.3,
2.3 Hz), 4.06–4.12 (1H, m), 4.16 (1H, dd, J = 12.3, 4.5 Hz), 4.89–
4.98 (2H, m), 5.39 (1H, t, J = 9.7 Hz), 5.68 (1H, d, J = 8.0 Hz), 6.71
(1H, d, J = 7.0 Hz), 6.83 (1H, d, J = 7.0 Hz), 6.92 (1H, s), 11.74 (1H,
s); MS m/z: 575 (M+H)+.
5.1.50. 4-(4-Benzyloxy-2-methylphenylmethyl)-5-isopropyl-1H-
5.1.55. 4-(2-Benzyloxyphenylmethyl)-5-ethyl-1H-pyrazol-3-yl
pyrazol-3-yl 2,3,4,6-tetra-O-acetyl-b-
D-glucopyranoside (10y)
b-D
-glucopyranoside (11e)
The title compound was prepared from 9y, as described for the
synthesis of 10a, in 22% yield.
The title compound was prepared from 10e, as described for the
synthesis of 11a, in 52% yield.
1H NMR (DMSO-d6) d: 1.03 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 1.81 (3H, s), 1.93 (3H, s), 1.98 (3H, s), 1.99 (3H, s),
2.21 (3H, s), 2.77 (1H, heptet, J = 7.0 Hz), 3.40 (1H, d, J = 16.4 Hz),
3.46 (1H, d, J = 16.4 Hz), 3.95 (1H, dd, J = 12.2, 2.1 Hz), 4.06–4.13
(1H, m), 4.17 (1H, dd, J = 12.2, 4.4 Hz), 4.90–4.99 (2H, m), 5.02
(2H, s), 5.39 (1H, t, J = 9.5 Hz), 5.68 (1H, d, J = 8.0 Hz), 6.67 (1H,
dd, J = 8.4, 2.5 Hz), 6.74 (1H, d, J = 8.4 Hz), 6.79 (1H, d, J = 2.5 Hz),
7.27–7.44 (5H, m), 11.73 (1H, s); MS m/z: 667 (M+H)+.
1H NMR (DMSO-d6) d: 0.93 (3H, t, J = 7.6 Hz), 2.32 (2H, q,
J = 7.6 Hz), 3.05–3.24 (4H, m), 3.40–3.49 (1H, m), 3.55–3.67 (3H,
m), 4.43 (1H, t, J = 5.8 Hz), 4.91 (1H, d, J = 4.8 Hz), 4.98 (1H, d,
J = 4.5 Hz), 5.10 (1H, d, J = 5.0 Hz), 5.15 (2H, s), 5.21 (1H, d,
J = 7.8 Hz), 6.78–6.84 (1H, m), 6.98–7.04 (2H, m), 7.07–7.14 (1H,
m), 7.30–7.36 (1H, m), 7.37–7.44 (2H, m), 7.45–7.51 (2H, m),
11.53 (1H, s); 13C NMR (100 MHz, CD3OD) d: 13.7, 19.2, 22.3,
62.7, 71.1, 71.3, 75.0, 78.0, 78.4, 102.5, 102.6, 112.8, 121.7, 128.1,
128.6, 128.9, 129.6, 130.8, 130.9, 139.0, 145.8, 157.6, 162.0; MS
m/z: 471 (M+H)+; HRMS (FAB+) calcd for C25H31N2O7 471.2126;
found 471.2124 (M+H)+.
5.1.51. 4-(4-Benzyloxyphenylmethyl)-5-trifluoromethyl-1H-
pyrazol-3-yl b-D-glucopyranoside (11a)
NaOMe (28% MeOH solution, 563 mg, 2.92 mmol) was added to
a solution of 10a (0.99 g, 1.46 mmol) in MeOH (6 mL), and the mix-
ture was stirred at room temperature for 21 h. The solvent was re-
moved under reduced pressure, and the residue was purified by
column chromatography on silica gel (eluent: CH2Cl2/
MeOH = 19:1–9:1–4:1) to give 11a (356 mg, 48%) as a white solid.
1H NMR (DMSO-d6) d: 3.08–3.27 (4H, m), 3.42–3.53 (1H, m),
3.60–3.76 (3H, m), 4.55 (1H, br s), 4.64–4.88 (1H, br), 4.92–5.17
(4H, m), 5.20–5.62 (1H, br), 6.89 (2H, d, J = 8.4 Hz), 7.09 (2H, d,
J = 8.4 Hz), 7.27–7.45 (5H, m), 13.27 (1H, br s); MS m/z: 509
(MꢁH)ꢁ. HRMS (FAB+) calcd for C24H26F3N2O7 511.1687; found
511.1688 (M+H)+.
5.1.56. 4-(2-Benzyloxyphenylmethyl)-5-isopropyl-1H-pyrazol-
3-yl b-D-glucopyranoside (11f)
The title compound was prepared from 10f, as described for the
synthesis of 11a, in 67% yield.
1H NMR (DMSO-d6) d: 1.00 (6H, d, J = 7.0 Hz), 2.77 (1H, heptet,
J = 7.0 Hz), 3.06–3.24 (4H, m), 3.40–3.49 (1H, m), 3.56–3.68 (3H,
m), 4.45 (1H, t, J = 5.9 Hz), 4.82–5.21 (5H, m), 5.22 (1H, d,
J = 7.5 Hz), 6.81 (1H, td, J = 7.5, 0.9 Hz), 6.98–7.03 (2H, m), 7.07–
7.14 (1H, m), 7.30–7.36 (1H, m), 7.38–7.44 (2H, m), 7.46–7.51
(2H, m), 11.53 (1H, br s); 13C NMR (100 MHz, CD3OD) d: 22.0,
22.1, 26.5, 62.8, 71.1, 71.3, 75.0, 78.0, 78.4, 101.9, 102.5, 112.8,
121.7, 128.0, 128.6, 128.9, 129.6, 130.7, 131.1, 139.0, 149.7,
157.5, 161.9; MS m/z: 485 (M+H)+; HRMS (FAB+) calcd for
5.1.52. 4-(3-Benzyloxyphenylmethyl)-5-trifluoromethyl-1H-
pyrazol-3-yl b-
D
-glucopyranoside (11b)
C
26H33N2O7 485.2282; found 485.2276 (M+H)+.
The title compound was prepared from 10b, as described for the
synthesis of 11a, in 42% yield.
5.1.57. 4-(2-Benzyloxyphenylmethyl)-5-tert-butyl-1H-pyrazol-
3-yl b- -glucopyranoside (11g)
1H NMR (DMSO-d6) d: 3.06–3.26 (4H, m), 3.40–3.53 (1H, m),
3.60–3.72 (1H, m), 3.76 (2H, s), 4.55 (1H, br s), 4.66–4.90 (1H,
br), 4.94–5.17 (4H, m), 5.18–5.62 (1H, br), 6.71–6.85 (3H, m),
7.16 (1H, t, J = 8.0 Hz), 7.28–7.45 (5H, m), 13.31 (1H, s); MS m/z:
509 (MꢁH)ꢁ. HRMS (FAB+) calcd for C24H26F3N2O7 511.1687;
found 511.1686 (M+H)+.
D
The title compound was prepared from 10g, as described for the
synthesis of 11a, in 68% yield.
1H NMR (DMSO-d6) d: 1.13 (9H, s), 3.04–3.23 (4H, m), 3.39–3.52
(1H, m), 3.59–3.66 (1H, m), 3.76 (2H, s), 4.43 (1H, t, J = 5.7 Hz), 4.92
(1H, d, J = 4.8 Hz), 4.97 (1H, d, J = 4.8 Hz), 5.08 (1H, d, J = 5.5 Hz),

N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
6609
5.17 (2H, s), 5.25 (1H, d, J = 8.0 Hz), 6.77–6.87 (2H, m), 7.01 (1H, d,
5.1.62. 4-(2-Trifluoromethylphenylmethyl)-5-isopropyl-1H-
pyrazol-3-yl b- -glucopyranoside (11l)
The title compound was prepared from 10l, as described for the
synthesis of 11a, in 65% yield.
J = 7.5 Hz), 7.07–7.14 (1H, m), 7.29–7.36 (1H, m), 7.38–7.44 (2H,
m), 7.47–7.52 (2H, m), 11.49 (1H, s); 13C NMR (100 MHz, CD3OD)
d: 22.7, 29.9, 33.4, 62.7, 71.1, 71.2, 74.9, 78.0, 78.3, 101.2, 102.4,
112.6, 121.6, 127.8, 128.5, 128.8, 129.5, 130.1, 131.5, 139.0,
150.9, 157.3, 163.1; MS m/z: 499 (M+H)+; HRMS (FAB+) calcd for
D
1H NMR (DMSO-d6) d: 1.035 (3H, d, J = 7.0 Hz), 1.040 (3H, d,
J = 7.0 Hz), 2.69 (1H, heptet, J = 7.0 Hz), 3.07–3.25 (4H, m), 3.45 (1H,
d, J = 11.4 Hz), 3.65 (1H, d, J = 11.4 Hz), 3.80 (1H, d, J = 17.4 Hz),
3.85 (1H, d, J = 17.4 Hz), 4.47 (1H, br s), 4.72–5.36 (4H, m), 7.23
(1H, d, J = 7.6 Hz), 7.37 (1H, t, J = 7.6 Hz), 7.54 (1H, t, J = 7.6 Hz),
7.67 (1H, d, J = 7.6 Hz), 11.73 (1H, br s); 13C NMR (100 MHz, DMSO-
d6) d: 21.4, 21.5, 22.9, 24.7, 60.7, 69.6, 73.2, 76.7, 77.3, 97.5, 100.3,
124.7 (q, J = 273 Hz), 125.3 (q, J = 6 Hz), 126.2, 126.4 (q, J = 29 Hz),
130.2, 132.4, 139.6, 147.0, 160.0; MS m/z: 447 (M+H)+; HRMS
(FAB+) calcd for C20H26F3N2O6 447.1738; found 447.1737 (M+H)+.
C
27H35N2O7 499.2439; found 499.2438 (M+H)+.
5.1.58. 4-(2-Benzyloxyphenylmethyl)-5-cyclopropyl-1H-
pyrazol-3-yl b- -glucopyranoside (11h)
D
The title compound was prepared from 10h, as described for the
synthesis of 11a, in 58% yield.
1H NMR (DMSO-d6) d: 0.55–0.67 (4H, m), 1.55–1.66 (1H, m),
3.05–3.23 (4H, m), 3.39–3.48 (1H, m), 3.57–3.73 (3H, m), 4.42
(1H, t, J = 5.8 Hz), 4.91 (1H, d, J = 4.8 Hz), 4.98 (1H, d, J = 4.8 Hz),
5.10 (1H, d, J = 5.0 Hz), 5.15 (2H, s), 5.20 (1H, d, J = 7.8 Hz), 6.82
(1H, td, J = 7.4, 1.0 Hz), 6.98–7.06 (2H, m), 7.08–7.15 (1H, m),
7.29–7.36 (1H, m), 7.37–7.43 (2H, m), 7.45–7.51 (2H, m), 11.33
(1H, br s); 13C NMR (100 MHz, CD3OD) d: 6.4, 6.5, 7.1, 22.4, 62.7,
71.1, 71.2, 75.0, 78.0, 78.3, 102.5, 104.1, 112.7, 121.6, 128.0,
128.6, 128.8, 129.5, 130.7, 130.9, 139.0, 145.4, 157.6, 162.0; MS
m/z: 483 (M+H)+; HRMS (FAB+) calcd for C26H31N2O7 483.2126;
found 483.2130 (M+H)+.
5.1.63. 4-(2-Ethylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl b-
D-glucopyranoside (11m)
The title compound was prepared from 10m, as described for
the synthesis of 11a, in 73% yield.
1H NMR (DMSO-d6) d: 1.03 (3H, d, J = 6.8 Hz), 1.05 (3H, d,
J = 6.8 Hz), 1.15 (3H, t, J = 7.5 Hz), 2.64–2.77 (3H, m), 3.06–3.23 (4H,
m), 3.40–3.49 (1H, m), 3.57–3.69 (3H, m), 4.44 (1H, t, J = 5.8 Hz),
4.92 (1H, br s), 4.99 (1H, br s), 5.07 (1H, d, J = 5.0 Hz), 5.19 (1H, d,
J = 7.8 Hz), 6.95 (1H, d, J = 7.3 Hz), 7.01–7.15 (3H, m), 11.57 (1H, br
s); 13C NMR (100 MHz, CD3OD) d: 15.3, 21.9, 22.0, 25.2, 26.7, 26.8,
62.8, 71.3, 75.0, 78.0, 78.4, 101.9, 102.6, 126.7, 127.2, 129.2, 129.7,
139.6, 143.2, 149.8, 161.7; MS m/z: 407 (M+H)+; HRMS (FAB+) calcd
for C21H31N2O6 407.2177; found 407.2175 (M+H)+.
5.1.59. 4-Benzyl-5-isopropyl-1H-pyrazol-3-yl b-D-
glucopyranoside (11i)
The title compound was prepared from 10i, as described for the
synthesis of 11a, in 43% yield.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 2.84 (1H, heptet, J = 7.0 Hz), 3.10–3.25 (4H, m), 3.41–
3.51 (1H, m), 3.55–3.68 (3H, m), 4.46 (1H, t, J = 5.7 Hz), 4.93 (1H,
d, J = 4.8 Hz), 5.00 (1H, d, J = 4.5 Hz), 5.15 (1H, d, J = 5.0 Hz), 5.22
(1H, d, J = 7.5 Hz), 7.08–7.26 (5H, m), 11.53 (1H, s); 13C NMR
(150 MHz, DMSO-d6) d: 21.57, 21.63, 24.6, 26.9, 60.6, 69.6, 73.3,
76.7, 77.2, 99.6, 100.1, 125.4, 128.0, 128.1, 141.7, 146.5, 159.5;
MS m/z: 379 (M+H)+; HRMS (FAB+) calcd for C19H27N2O6
379.1864; found 379.1869 (M+H)+.
5.1.64. 4-(2-Methoxyphenylmethyl)-5-isopropyl-1H-pyrazol-3-
yl b-D-glucopyranoside (11n)
The title compound was prepared from 10n, as described for the
synthesis of 11a, in 55% yield.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 7.0 Hz), 1.08 (3H, d,
J = 7.0 Hz), 2.82 (1H, heptet, J = 7.0 Hz), 3.06–3.24 (4H, m), 3.39–
3.67 (4H, m), 3.80 (3H, s), 4.45 (1H, br s), 4.78–5.19 (3H, m), 5.21
(1H, d, J = 7.8 Hz), 6.77–6.83 (1H, m), 6.91 (1H, d, J = 7.8 Hz), 6.97
(1H, dd, J = 7.4, 1.4 Hz), 7.09–7.16 (1H, m), 11.54 (1H, br s); 13C
NMR (150 MHz, CD3OD) d: 21.9, 22.0, 26.5, 55.8, 62.8, 71.3, 75.0,
78.0, 78.4, 101.8, 102.5, 111.1, 121.3, 128.1, 130.4, 130.6, 149.8,
158.5, 161.9; MS m/z: 409 (M+H)+; HRMS (FAB+) calcd for
5.1.60. 4-(2-Fluorophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
b-
D-glucopyranoside (11j)
The title compound was prepared from 10j, as described for the
synthesis of 11a, in 78% yield.
C
20H29N2O7 409.1969; found 409.1977 (M+H)+.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 7.0 Hz), 1.08 (3H, d,
J = 7.0 Hz), 2.84 (1H, heptet, J = 7.0 Hz), 3.09–3.25 (4H, m), 3.40–
3.51 (1H, m), 3.57–3.70 (3H, m), 4.47 (1H, t, J = 5.9 Hz), 4.86–5.28
(4H, m), 7.04–7.14 (2H, m), 7.15–7.23 (2H, m), 11.60 (1H, br s);
13C NMR (100 MHz, CD3OD) d: 20.7 (d, J = 4 Hz), 22.0, 26.6, 62.8,
71.3, 75.0, 78.1, 78.4, 101.0, 102.6, 115.7 (d, J = 22 Hz), 125.1 (d,
J = 4 Hz), 128.7 (d, J = 8 Hz), 129.4 (d, J = 15 Hz), 131.8 (d, J = 4 Hz),
149.7, 161.8, 162.1 (d, J = 242 Hz); MS m/z: 397 (M+H)+; HRMS
(FAB+) calcd for C19H26FN2O6 397.1769; found 397.1777 (M+H)+.
5.1.65. 4-(2-Cyanophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
b-D
-glucopyranoside (11o)
The title compound was prepared from 10o, as described for the
synthesis of 11a, in 58% yield.
1H NMR (DMSO-d6) d: 1.07 (3H, d, J = 6.9 Hz), 1.08 (3H, d,
J = 6.9 Hz), 2.87 (1H, heptet, J = 6.9 Hz), 3.09–3.25 (4H, m), 3.41–
3.50 (1H, m), 3.60–3.68 (1H, m), 3.81 (1H, d, J = 16.6 Hz), 3.86
(1H, d, J = 16.6 Hz), 4.46 (1H, t, J = 5.8 Hz), 4.94 (1H, d, J = 4.8 Hz),
5.01 (1H, d, J = 4.8 Hz), 5.13 (1H, d, J = 5.3 Hz), 5.22 (1H, d,
J = 7.8 Hz), 7.33–7.41 (2H, m), 7.59 (1H, td, J = 7.7, 1.2 Hz), 7.76
(1H, dd, J = 7.5, 1.2 Hz), 11.68 (1H, s); 13C NMR (100 MHz, CD3OD)
d: 22.0, 22.1, 26.7, 26.8, 62.8, 71.3, 74.9, 78.0, 78.4, 100.6, 102.4,
112.8, 119.1, 127.9, 131.0, 133.6, 134.2, 146.6, 149.7, 161.7; MS
m/z: 404 (M+H)+; HRMS (FAB+) calcd for C20H26N3O6 404.1816;
found 404.1820 (M+H)+.
5.1.61. 4-(2-Methylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
b-
D-glucopyranoside (11k)
The title compound was prepared from 10k, as described for the
synthesis of 11a, in 62% yield.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 2.30 (3H, s), 2.74 (1H, heptet, J = 7.0 Hz), 3.05–3.23
(4H, m), 3.40–3.48 (1H, m), 3.52–3.65 (3H, m), 4.44 (1H, t,
J = 5.7 Hz), 4.92 (1H, d, J = 4.5 Hz), 4.98 (1H, d, J = 4.5 Hz), 5.08
(1H, d, J = 5.3 Hz), 5.18 (1H, d, J = 7.5 Hz), 6.90–6.96 (1H, m),
7.01–7.13 (3H, m), 11.58 (1H, br s); 13C NMR (100 MHz, CD3OD)
d: 19.8, 21.9, 22.0, 25.9, 26.6, 62.7, 71.3, 74.9, 78.0, 78.4, 101.5,
102,7, 126.7, 127.0, 129.2, 130.9, 137.3, 140.3, 149.8, 161.7; MS
m/z: 393 (M+H)+; HRMS (FAB+) calcd for C20H29N2O6 393.2020;
found 393.2022 (M+H)+.
5.1.66. 4-(4-Fluorophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
b-D
-glucopyranoside (11p)
The title compound was prepared from 10p, as described for the
synthesis of 11a, in 60% yield.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 2.85 (1H, heptet, J = 7.0 Hz), 3.10–3.26 (4H, m), 3.42–
3.52 (1H, m), 3.54–3.69 (3H, m), 4.48 (1H, t, J = 5.5 Hz), 4.94 (1H,

6610
N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
br s), 5.02 (1H, br s), 5.14–5.24 (2H, m), 7.00–7.08 (2H, m), 7.21–
7.28 (2H, m), 11.56 (1H, br s); 13C NMR (100 MHz, DMSO-d6) d:
21.6, 21.7, 24.6, 26.1, 60.7, 69.6, 73.3, 76.7, 77.3, 99.8, 100.3,
114.6 (d, J = 21 Hz), 129.9 (d, J = 7 Hz), 137.9 (d, J = 3 Hz), 146.6,
159.4, 160.4 (d, J = 239 Hz); MS m/z: 397 (M+H)+; HRMS (FAB+)
calcd for C19H26FN2O6 397.1769; found 397.1770 (M+H)+.
3.52 (1H, m), 3.60–3.78 (3H, m), 4.49 (1H, t, J = 5.7 Hz), 4.94 (1H,
d, J = 4.8 Hz), 5.02 (1H, d, J = 4.8 Hz), 5.17–5.29 (2H, m), 7.43 (2H,
d, J = 8.3 Hz), 7.70 (2H, d, J = 8.3 Hz), 11.63 (1H, br s); 13C NMR
(100 MHz, DMSO-d6) d: 21.66, 21.71, 24.6, 27.1, 60.7, 69.6, 73.3,
76.7, 77.3, 98.8, 100.4, 108.3, 119.1, 129.3, 132.0, 146.8, 147.8,
159.5; MS m/z: 402 (MꢁH)ꢁ; HRMS (FAB+) calcd for C20H26N3O6
404.1816; found 404.1817 (M+H)+.
5.1.67. 4-(4-Methylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
b-D
-glucopyranoside (11q)
5.1.72. 4-(3-Methylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
The title compound was prepared from 10q, as described for the
b-D
-glucopyranoside (11v)
synthesis of 11a, in 53% yield.
The title compound was prepared from 10v, as described for the
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 6.9 Hz), 1.06 (3H, d,
J = 6.9 Hz), 2.23 (3H, s), 2.83 (1H, heptet, J = 6.9 Hz), 3.09–3.26
(4H, m), 3.41–3.69 (4H, m), 4.46 (1H, t, J = 5.6 Hz), 4.93 (1H, d,
J = 4.5 Hz), 5.00 (1H, d, J = 4.8 Hz), 5.14 (1H, d, J = 5.3 Hz), 5.22
(1H, d, J = 7.5 Hz), 7.03 (2H, d, J = 8.0 Hz), 7.08 (2H, d, J = 8.0 Hz),
11.51 (1H, br s); 13C NMR (100 MHz, DMSO-d6) d: 20.5, 21.57,
21.63, 24.6, 26.4, 60.6, 69.6, 73.2, 76.7, 77.2, 99.7, 100.1, 127.9,
128.5, 134.1, 138.5, 146.4, 159.5; MS m/z: 393 (M+H)+; HRMS
(FAB+) calcd for C20H29N2O6 393.2020; found 393.2030 (M+H)+.
synthesis of 11a, in 62% yield.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.07 (3H, d,
J = 7.0 Hz), 2.24 (3H, s), 2.83 (1H, heptet, J = 7.0 Hz), 3.10–3.26
(4H, m), 3.42–3.51 (1H, m), 3.53–3.68 (3H, m), 4.46 (1H, t,
J = 5.8 Hz), 4.93 (1H, d, J = 4.8 Hz), 5.00 (1H, d, J = 4.5 Hz), 5.13
(1H, d, J = 5.3 Hz), 5.23 (1H, d, J = 7.5 Hz), 6.93 (1H, d, J = 7.5 Hz),
6.98 (1H, d, J = 7.5 Hz), 7.02 (1H, s), 7.11 (1H, t, J = 7.5 Hz), 11.52
(1H, br s); 13C NMR (100 MHz, DMSO-d6) d: 21.0, 21.6, 21.7, 24.6,
26.8, 60.6, 69.6, 73.3, 76.8, 77.2, 99.6, 100.0, 125.2, 126.1, 127.9,
128.8, 137.0, 141.6, 146.5, 159.5; MS m/z: 393 (M+H)+; HRMS
(FAB+) calcd for C20H29N2O6 393.2020; found 393.2022 (M+H)+.
5.1.68. 4-(4-Ethylphenylmethyl)-5-isopropyl-1H-pyrazol-3-yl b-
D-glucopyranoside (11r)
The title compound was prepared from 10r, as described for the
5.1.73. 4-(3-Benzyloxyphenylmethyl)-5-isopropyl-1H-pyrazol-
synthesis of 11a, in 54% yield.
3-yl b-D-glucopyranoside (11w)
1H NMR (DMSO-d6) d: 1.06 (3H, d, J = 7.0 Hz), 1.07 (3H, d,
J = 7.0 Hz), 1.13 (3H, t, J = 7.5 Hz), 2.53 (2H, q, J = 7.5 Hz), 2.84 (1H,
heptet, J = 7.0 Hz), 3.08–3.26 (4H, m), 3.40–3.68 (4H, m), 4.46 (1H,
t, J = 5.8 Hz), 4.92 (1H, d, J = 4.5 Hz), 4.99 (1H, d, J = 4.5 Hz), 5.14
(1H, d, J = 5.0 Hz), 5.22 (1H, d, J = 7.5 Hz), 7.03–7.13 (4H, m), 11.51
(1H, s); 13C NMR (100 MHz, CD3OD) d: 16.3, 21.99, 22.0, 26.6,
27.8, 29.5, 62.8, 71.3, 75.0, 78.0, 78.4, 102.6, 102.8, 128.7, 129.3,
140.1, 142.9, 149.6, 161.7; MS m/z: 407 (M+H)+; HRMS (FAB+) calcd
for C21H31N2O6 407.2177; found 407.2172 (M+H)+.
The title compound was prepared from 10w, as described for
the synthesis of 11a, in 74% yield.
1H NMR (DMSO-d6) d: 1.04 (3H, d, J = 6.9 Hz), 1.06 (3H, d,
J = 6.9 Hz), 2.83 (1H, heptet, J = 6.9 Hz), 3.10–3.26 (4H, m), 3.40–
3.50 (1H, m), 3.53–3.68 (3H, m), 4.46 (1H, t, J = 5.7 Hz), 4.87–5.10
(4H, m), 5.16 (1H, d, J = 3.5 Hz), 5.24 (1H, d, J = 7.5 Hz), 6.74–6.81
(2H, m), 6.82–6.87 (1H, m), 7.14 (1H, t, J = 7.9 Hz), 7.28–7.45 (5H,
m), 11.53 (1H, br s); MS m/z: 485 (M+H)+.
5.1.74. 4-(2,4-Dimethylphenylmethyl)-5-isopropyl-1H-pyrazol-
5.1.69. 4-(4-Methoxyphenylmethyl)-5-isopropyl-1H-pyrazol-3-
3-yl b-D-glucopyranoside (11x)
yl b-
D
-glucopyranoside (11s)
The title compound was prepared from 10x, as described for the
synthesis of 11a, in 57% yield.
The title compound was prepared from 10s, as described for the
synthesis of 11a, in 53% yield.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 2.20 (3H, s), 2.25 (3H, s), 2.73 (1H, heptet, J = 7.0 Hz),
3.06–3.23 (4H, m), 3.40–3.66 (4H, m), 4.45 (1H, t, J = 5.8 Hz), 4.92
(1H, d, J = 4.5 Hz), 4.98 (1H, d, J = 4.5 Hz), 5.08 (1H, d, J = 5.3 Hz),
5.18 (1H, d, J = 7.8 Hz), 6.80 (1H, d, J = 8.0 Hz), 6.90 (1H, d,
J = 8.0 Hz), 6.92 (1H, s), 11.56 (1H, br s); 13C NMR (100 MHz,
DMSO-d6) d: 19.2, 20.4, 21.6, 21.7, 23.9, 24.6, 60.6, 69.6, 73.2,
76.7, 77.2, 98.4, 100.1, 126.0, 127.8, 130.3, 134.2, 135.2, 136.1,
146.8, 159.7; MS m/z: 407 (M+H)+; HRMS (FAB+) calcd for
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 6.9 Hz), 1.06 (3H, d,
J = 6.9 Hz), 2.84 (1H, heptet, J = 6.9 Hz), 3.10–3.26 (4H, m), 3.42–
3.68 (4H, m), 3.69 (3H, s), 4.46 (1H, t, J = 5.8 Hz), 4.93 (1H, d,
J = 4.5 Hz), 5.00 (1H, d, J = 4.5 Hz), 5.15 (1H, d, J = 5.3 Hz), 5.22
(1H, d, J = 7.8 Hz), 6.76–6.81 (2H, m), 7.08–7.14 (2H, m), 11.50
(1H, s); 13C NMR (100 MHz, CD3OD) d: 22.00, 22.01, 26.6, 27.3,
55.7, 62.8, 71.3, 75.0, 78.0, 78.4, 102.6, 103.0, 114.7, 130.2, 134.9,
149.4, 159.3, 161.7; MS m/z: 409 (M+H)+; HRMS (FAB+) calcd for
C
20H29N2O7 409.1969; found 409.1972 (M+H)+.
C
21H31N2O6 407.2177; found 407.2179 (M+H)+.
5.1.70. 4-(4-Benzyloxyphenylmethyl)-5-isopropyl-1H-pyrazol-
3-yl b- -glucopyranoside (11t)
5.1.75. 4-(4-Benzyloxy-2-methylphenylmethyl)-5-isopropyl-1H-
pyrazol-3-yl b- -glucopyranoside (11y)
D
D
The title compound was prepared from 10t, as described for the
synthesis of 11a, in 68% yield.
The title compound was prepared from 10y, as described for the
synthesis of 11a, in 73% yield.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 6.9 Hz), 1.06 (3H, d,
J = 6.9 Hz), 2.83 (1H, heptet, J = 6.9 Hz), 3.09–3.27 (4H, m), 3.42–
3.68 (4H, m), 4.47 (1H, t, J = 5.6 Hz), 4.93 (1H, d, J = 4.8 Hz), 5.00
(1H, d, J = 4.5 Hz), 5.03 (2H, s), 5.15 (1H, d, J = 5.0 Hz), 5.22 (1H,
d, J = 7.3 Hz), 6.87 (2H, d, J = 8.5 Hz), 7.12 (2H, d, J = 8.5 Hz), 7.28–
7.45 (5H, m), 11.51 (1H, s); MS m/z: 485 (M+H)+.
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 2.26 (3H, s), 2.73 (1H, heptet, J = 7.0 Hz), 3.06–3.23
(4H, m), 3.40–3.56 (3H, m), 3.58–3.66 (1H, m), 4.44 (1H, t,
J = 5.8 Hz), 4.91 (1H, d, J = 4.5 Hz), 4.98 (1H, d, J = 4.5 Hz), 5.02
(2H, s), 5.08 (1H, d, J = 5.3 Hz), 5.19 (1H, d, J = 7.8 Hz), 6.70 (1H,
dd, J = 8.3, 2.8 Hz), 6.79 (1H, d, J = 2.8 Hz), 6.84 (1H, d, J = 8.3 Hz),
7.28–7.44 (5H, m), 11.55 (1H, s); MS m/z: 499 (M+H)+.
5.1.71. 4-(4-Cyanophenylmethyl)-5-isopropyl-1H-pyrazol-3-yl
b-D
-glucopyranoside (11u)
5.1.76. 4-Benzyl-5-trifluoromethyl-1H-pyrazol-3-yl b-D-
The title compound was prepared from 10u, as described for the
glucopyranoside (5)
synthesis of 11a, in 38% yield.
The title compound was prepared from 4-benzyl-5-trifluoro-
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 6.9 Hz), 1.06 (3H, d,
J = 6.9 Hz), 2.86 (1H, heptet, J = 6.9 Hz), 3.10–3.27 (4H, m), 3.41–
methyl-1,2-dihydro-3H-pyrazol-3-one24 via 4-benzyl-5-trifluoro-
methyl-1H-pyrazol-3-yl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranoside,

N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
6611
as described for the synthesis of 10a and 11a, in 8% and 48% yield,
respectively.
3.56 (3H, m), 3.59–3.68 (1H, m), 4.46 (1H, t, J = 5.8 Hz), 4.93 (1H,
d, J = 4.5 Hz), 5.00 (1H, d, J = 4.5 Hz), 5.14 (1H, d, J = 5.0 Hz), 5.21
(1H, d, J = 7.3 Hz), 6.61 (2H, d, J = 8.5 Hz), 6.98 (2H, d, J = 8.5 Hz),
9.04 (1H, s), 11.48 (1H, s); 13C NMR (100 MHz, DMSO-d6) d: 21.6,
21.7, 24.6, 26.1, 60.6, 69.6, 73.3, 76.8, 77.2, 100.1, 100.3, 114.8,
128.9, 131.8, 146.4, 155.0, 159.5; MS m/z: 395 (M+H)+; HRMS
(FAB+) calcd for C19H27N2O7 395.1813; found 395.1816 (M+H)+.
1H NMR (DMSO-d6) d: 3.08–3.25 (4H, m), 3.48 (1H, d,
J = 10.2 Hz), 3.67 (1H, d, J = 10.2 Hz), 3.79 (2H, s), 4.54 (1H, br s),
4.62–5.00 (1H, br), 5.03 (1H, br s), 5.12 (1H, br s), 5.25–5.62 (1H,
br), 7.08–7.28 (5H, m), 13.31 (1H, br s); MS m/z: 403 (MꢁH)ꢁ;
HRMS (FAB+) calcd for C17H20F3N2O6 405.1268; found 405.1270
(M+H)+.
5.1.82. 4-(3-Hydroxyphenylmethyl)-5-isopropyl-1H-pyrazol-3-
5.1.77. 5-Trifluoromethyl-4-(4-hydroxyphenylmethyl)-1H-
yl b-D-glucopyranoside (12f)
pyrazol-3-yl b-
D
-glucopyranoside (12a)
The title compound was prepared from 11w, as described for
the synthesis of 12a, in 86% yield.
To a solution of 11a (100 mg, 0.196 mmol) in MeOH (6 mL), 10%
Pd–C (50% wet with water, 30 mg) was added. The mixture was
stirred under a hydrogen atmosphere at room temperature over-
night. Insoluble materials were removed by filtration and washed
with MeOH. The filtrate and washing were concentrated under re-
duced pressure, and the residue was purified by column chroma-
tography on silica gel (eluent: CH2Cl2/MeOH = 9:1–3:1) to give
12a (70 mg, 85%) as a white solid.
1H NMR (DMSO-d6) d: 1.07 (6H, d, J = 7.0 Hz), 2.82 (1H, heptet,
J = 7.0 Hz), 3.10–3.26 (4H, m), 3.43–3.61 (3H, m), 3.65 (1H, d,
J = 11.3 Hz), 4.55 (1H, br s), 4.75–5.20 (3H, m), 5.21 (1H, d,
J = 7.5 Hz), 6.51 (1H, dd, J = 7.9, 1.8 Hz), 6.57–6.60 (1H, m), 6.63
(1H, d, J = 7.9 Hz), 7.01 (1H, t, J = 7.9 Hz), 9.16 (1H, br s), 11.54
(1H, br s); 13C NMR (100 MHz, DMSO-d6) d: 21.6, 21.7, 24.7, 26.8,
60.6, 69.6, 73.3, 76.7, 77.2, 99.6, 100.3, 112.5, 115.0, 118.8, 128.9,
143.1, 146.7, 157.1, 159.5; MS m/z: 395 (M+H)+; HRMS (FAB+)
calcd for C19H27N2O7 395.1813; found 395.1816 (M+H)+.
1H NMR (DMSO-d6) d: 3.10–3.25 (4H, m), 3.48 (1H, dd, J = 11.9,
5.1 Hz), 3.62–3.70 (3H, m), 4.35–5.20 (4H, m), 5.20–5.70 (1H, br),
6.60–6.65 (2H, m), 6.96 (2H, d, J = 8.5 Hz), 9.15 (1H, s), 12.80–
13.70 (1H, br); MS m/z: 419 (MꢁH)ꢁ; HRMS (FAB+) calcd for
5.1.83. 4-(4-Hydroxy-2-methylphenylmethyl)-5-isopropyl-1H-
pyrazol-3-yl b-D-glucopyranoside (12g)
C
17H20F3N2O7 421.1217; found 421.1222 (M+H)+.
The title compound was prepared from 11y, as described for the
synthesis of 12a, in 85% yield.
5.1.78. 5-Trifluoromethyl-4-(3-hydroxyphenylmethyl)-1H-
pyrazol-3-yl b- -glucopyranoside (12b)
The title compound was prepared from 11b, as described for the
synthesis of 12a, in 80% yield.
D
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 2.20 (3H, s), 2.73 (1H, heptet, J = 7.0 Hz), 3.06–3.25
(4H, m), 3.39–3.53 (3H, m), 3.58–3.67 (1H, m), 4.46 (1H, t,
J = 5.8 Hz), 4.93 (1H, d, J = 4.5 Hz), 5.00 (1H, d, J = 4.5 Hz), 5.09
(1H, d, J = 5.0 Hz), 5.19 (1H, d, J = 7.8 Hz), 6.44 (1H, dd, J = 8.3,
2.4 Hz), 6.53 (1H, d, J = 2.4 Hz), 6.72 (1H, d, J = 8.3 Hz), 8.95 (1H,
s), 11.53 (1H, s); 13C NMR (100 MHz, DMSO-d6) d: 19.4, 21.6,
21.7, 23.5, 24.6, 60.6, 69.6, 73.3, 76.7, 77.2, 98.9, 100.0, 112.1,
116.5, 128.9, 129.3, 136.4, 146.7, 155.0, 159.7; MS m/z: 409
(M+H)+; HRMS (FAB+) calcd for C20H29N2O7 409.1969; found
409.1970 (M+H)+.
1H NMR (DMSO-d6) d: 3.10–3.24 (4H, m), 3.49 (1H, dd, J = 11.8,
5.0 Hz), 3.63–3.75 (3H, m), 4.40–5.20 (4H, m), 5.20–5.75 (1H, br),
6.51–6.57 (2H, m), 6.60 (1H, d, J = 7.7 Hz), 7.03 (1H, dd, J = 8.7,
7.7 Hz), 9.22 (1H, br s), 12.80–13.80 (1H, br); MS m/z: 419
(MꢁH)ꢁ; HRMS (FAB+) calcd for C17H20F3N2O7 421.1217; found
421.1215 (M+H)+.
5.1.79. 5-Trifluoromethyl-4-(2-hydroxyphenylmethyl)-1H-
pyrazol-3-yl b-D-glucopyranoside (12c)
The title compound was prepared from 11c, as described for the
synthesis of 12a, in 77% yield.
5.1.84. X-ray crystallographic analysis of compound 10a
A single crystal (1.0 ꢂ 0.20 ꢂ 0.16 mm) of 10a was obtained by
recrystallization from EtOAc/hexane. Reflection data were collected
using a Rigaku AFC7R diffractometer with graphite monochromated
1H NMR (DMSO-d6) d: 3.09–3.20 (4H, m), 3.42–3.52 (1H, m),
3.65 (1H, d, J = 11.8 Hz), 3.71 (2H, s), 4.30–5.70 (5H, m), 6.61–
6.73 (2H, m), 6.77 (1H, d, J = 7.8 Hz), 6.94–7.00 (1H, m), 9.42 (1H,
br s), 12.80–13.70 (1H, br); MS m/z: 419 (MꢁH)ꢁ; HRMS (FAB+)
calcd for C17H20F3N2O7 421.1217; found 421.1222 (M+H)+.
Mo Ka radiation. The structure was determined by direct methods
(SIR92) and refined using the full-matrix least-squares technique
(CrystalStructure: Rigaku) with anisotropic temperature factors for
the non-hydrogen atoms. Hydrogen atoms were included using a rid-
ing model. The crystal contained four molecules in the asymmetric
unit. Crystal data: C32H33F3N2O11; M = 678.61; monoclinic; space
5.1.80. 4-(2-Hydroxyphenylmethyl)-5-isopropyl-1H-pyrazol-3-
yl b-D-glucopyranoside (12d)
The title compound was prepared from 11f, as described for the
synthesis of 12a, in 86% yield.
group P212121 (#19); cell constants
Å, c = 5.911(3) Å, = b =
= 90.000(7)°, V = 3382.86(19) Å3; Z = 4;
Dc = 1.332 g/cm3; unique reflections, 4431; observed reflections [I
>3 (I)], 3682; R1 = 0.049, wR2 = 0.148; GOF = 0.932.
a = 19.705(4) Å, b = 29.043(7)
a
c
1H NMR (DMSO-d6) d: 1.06 (3H, d, J = 7.0 Hz), 1.07 (3H, d,
J = 7.0 Hz), 2.83 (1H, heptet, J = 7.0 Hz), 3.08–3.24 (4H, m), 3.41–
3.67 (4H, m), 4.45 (1H, br s), 4.82–5.18 (3H, m), 5.22 (1H, d,
J = 7.8 Hz), 6.64 (1H, td, J = 7.6, 1.2 Hz), 6.75 (1H, dd, J = 7.6, 1.2 Hz),
6.87 (1H, dd, J = 7.6, 1.5 Hz), 6.94 (1H, td, J = 7.6, 1.5 Hz), 9.34 (1H,
br s), 11.52 (1H, br s); 13C NMR (100 MHz, CD3OD) d: 21.8, 22.1,
26.5, 62.8, 71.3, 75.0, 78.0, 78.4, 102.0, 102.7, 115.6, 120.5, 127.8,
128.8, 130.6, 149.8, 155.9, 161.8; MS m/z: 395 (M+H)+; HRMS
(FAB+) calcd for C19H27N2O7 395.1813; found 395.1816 (M+H)+.
r
5.1.85. Solubility determination
Compounds were incubated in distilled water, and precipitates
were separated by filtration. The solubility was determined by
high-performance liquid chromatography (HPLC) analysis of each
filtrate.
5.2. Biology
5.1.81. 4-(4-Hydroxyphenylmethyl)-5-isopropyl-1H-pyrazol-3-
yl b-
D
-glucopyranoside (12e)
5.2.1. SGLT1 and SGLT2 inhibition assay
The title compound was prepared from 11t, as described for the
synthesis of 12a, in 87% yield.
Human and rat SGLT expression plasmids were constructed as
reported previously.15,16 Cell culture, transfection procedure, and
1H NMR (DMSO-d6) d: 1.05 (3H, d, J = 7.0 Hz), 1.06 (3H, d,
J = 7.0 Hz), 2.82 (1H, heptet, J = 7.0 Hz), 3.09–3.26 (4H, m), 3.42–
[
14C]-AMG uptake experiments were performed as reported previ-
ously.16 In the experiments, 1 mM [14C]-AMG concentration in the

6612
N. Fushimi et al. / Bioorg. Med. Chem. 20 (2012) 6598–6612
uptake buffer was used. The concentration of the test compounds
required to inhibit 50% uptake of [14C]-AMG (IC50) was calculated
using logit plot. Phlorizin (6) was always included in the experi-
ments as a reference standard.
unobu Mukaiyama and Dr. Akihito Hirabayashi for providing valu-
able suggestions during the preparation of this manuscript.
Supplementary data
5.2.2. Oral mixed carbohydrate tolerance tests in NA-STZ rats
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2012.09.037.
5.2.2.1. Preparation of diabetic rat model.
Male Wistar rats,
aged 8 weeks, were injected intraperitoneally with nicotinamide
(230 mg/kg). Fifteen minutes after injection, these rats were in-
jected intravenously with streptozotocin (85 mg/kg) from the tail
vein under anesthesia with ether. After 1 week, the rats were
starved overnight, and a glucose tolerance test (2 g/kg) was per-
formed. Rats, which displayed greater than 250 mg/dL plasma glu-
cose concentration at 1 h after glucose load, were selected for use
in subsequent tests.
References and notes
1. American Diabetes Association Diabetes Care 2012, 35, S64.
2. Whiting, D. R.; Guariguata, L.; Weil, C.; Shaw, J. Diabetes Res. Clin. Pract. 2011,
94, 311.
3. (a) Fava, S. Expert Rev. Cardiovasc. Ther. 2008, 6, 859; (b) Davidson, J. Diabetes
Care 1919, 2003, 26; (c) Tominaga, M.; Eguchi, H.; Manaka, H.; Igarashi, K.;
Kato, T.; Sekikawa, A. Diabetes Care 1999, 22, 920; (d) The DECODE Study Group
Lancet 1999, 354, 617.
4. (a) Chiasson, J.-L.; Josse, R. G.; Gomis, R.; Hanefeld, M.; Karasik, A.; Laakso, M.
Lancet 2002, 359, 2072; (b) Chiasson, J.-L.; Josse, R. G.; Gomis, R.; Hanefeld, M.;
Karasik, A.; Laakso, M. JAMA 2003, 290, 486.
5.2.2.2. Mixed carbohydrate tolerance test.
After being
starved overnight, the diabetic rats were orally administered test
compounds suspended in 0.5% carboxymethyl cellulose (CMC), or
0.5% CMC alone (vehicle group). Immediately after test compound
administration, 2 g/kg of mixed carbohydrate (starch/sucrose/lac-
tose = 6:3:1)25 was loaded orally. The blood was collected from
the tail artery immediately before and after administration and
treated instantly with heparin. The blood was centrifuged, and
the plasma was collected to quantify the plasma glucose concen-
tration using the glucose oxidase method. Dunnett’s multiple com-
parison (two-sided) was used to estimate the degree of statistical
significance of the difference between the vehicle and individual
groups.
5. Scott, L. J.; Spencer, C. M. Drugs 2000, 59, 521.
6. Levin, R. J. Am. J. Clin. Nutr. 1994, 59, 690S.
7. Wright, E. M.; Hirayama, B. A.; Loo, D. F. J. Intern. Med. 2007, 261, 32.
8. Turk, E.; Zabel, B.; Mundlos, S.; Dyer, J.; Wright, E. M. Nature 1991, 350, 354.
9. Dyer, J.; Wood, I. S.; Palejwala, A.; Ellis, A.; Shirazi-Beechey, S. P. Am. J. Physiol.
Gastrointest. Liver Physiol. 2002, 282, G241.
10. (a) Miyamoto, K.; Hase, K.; Taketani, Y.; Minami, H.; Oka, T.; Nakabou, Y.;
Hagihira, H. Biochem. Biophys. Res. Commun. 1991, 181, 1110; (b) Dyer, J.;
Garner, A.; Wood, I. S.; Sharma, A. K.; Chandranath, I.; Shirazi-Beechey, S. P.
Biochem. Soc. Trans. 1997, 25, 479S.
11. (a) Isaji, M. Curr. Opin. Investig. Drugs 2007, 8, 285; (b) Handlon, A. L. Expert
Opin. Ther. Patents 2005, 15, 1531; (c) Washburn, W. N. Expert Opin. Ther.
Patents 2009, 19, 1485; (d) Washburn, W. N. Expert Opin. Ther. Patents 2012, 22,
483.
12. (a) Takii, H.; Matsumoto, K.; Kometani, T.; Okada, S.; Fushiki, T. Biosci. Biotech.
Biochem. 1997, 61, 1531; (b) Ikumi, Y.; Kida, T.; Sakuma, S.; Yamashita, S.;
Akashi, M. J. Control. Release 2008, 125, 42; (c) Sakuma, S.; Kanamitsu, S.;
Teraoka, Y.; Masaoka, Y.; Kataoka, M.; Yamashita, S.; Shirasaka, Y.; Tamai, I.;
Muraoka, M.; Nakatsuji, Y.; Kida, T.; Akashi, M. Mol. Pharm. 2012, 9, 922.
13. Kanai, Y.; Lee, W.-S.; You, G.; Brown, D.; Hediger, M. A. J. Clin. Invest. 1994, 93,
397.
14. (a) Isaji, M. Kidney Int. 2011, 79, S14; (b) Jabbour, S. A.; Goldstein, B. J. Int. J. Clin.
Pract. 2008, 62, 1279; (c) DeFronzo, R. A.; Davidson, J. A.; Prato, S. D. Diabetes
Obes. Metab. 2012, 14, 5.
5.2.3. Metabolic stability
Test compounds (final concentration, 50 lM) were incubated in
the assay buffer containing 0.8 mg/mL rat intestinal microsomes,
1 mg/mL b-nicotinamide-adenine-dinucleotide phosphate, 50 mM
ammonium acetate buffer (pH 5.0) and 10 mM magnesium chlo-
ride at 37 °C. Concentrations of the test compounds were deter-
mined by LC–MS. Metabolic stability was calculated from the
ratio of the test compound concentration at 0 min to its concentra-
tion after 30 and 60 min incubation.
15. Katsuno, K.; Fujimori, Y.; Takemura, Y.; Hiratochi, M.; Itoh, F.; Komatsu, Y.;
Fujikura, H.; Isaji, M. J. Pharmacol. Exp. Ther. 2007, 320, 323.
16. Fujimori, Y.; Katsuno, K.; Nakashima, I.; Ishikawa-Takemura, Y.; Fujikura, H.;
Isaji, M. J. Pharmacol. Exp. Ther. 2008, 327, 268.
17. Ehrenkranz, J. R. L.; Lewis, N. G.; Kahn, C. R.; Roth, J. Diabetes Metab. Res. Rev.
2005, 21, 31.
18. (a) Hongu, M.; Saito, K.; Tsujihara, K. Synth. Commun. 1999, 29, 2775; (b)
Zenkoh, T.; Tanaka, H.; Setoi, H.; Takahashi, T. Synlett 2002, 867.
19. Tsujihara, K.; Hongu, M.; Saito, K.; Inamasu, M.; Arakawa, K.; Oku, A.;
Matsumoto, M. Chem. Pharm. Bull. 1996, 44, 1174.
20. Meng, W.; Ellsworth, B. A.; Nirschl, A. A.; McCann, P. J.; Patel, M.; Girotra, R. N.;
Wu, G.; Sher, P. M.; Morrison, E. P.; Biller, S. A.; Zahler, R.; Deshpande, P. P.;
Pullockaran, A.; Hagan, D. L.; Morgan, N.; Taylor, J. R.; Obermeier, M. T.;
Humphreys, W. G.; Khanna, A.; Discenza, L.; Robertson, J. G.; Wang, A.; Han, S.;
Wetterau, J. R.; Janovitz, E. B.; Flint, O. P.; Whaley, J. M.; Washburn, W. N. J. Med.
Chem. 2008, 51, 1145.
21. Xu, B.; Feng, Y.; Cheng, H.; Song, Y.; Lv, B.; Wu, Y.; Wang, C.; Li, S.; Xu, M.; Du, J.;
Peng, K.; Dong, J.; Zhang, W.; Zhang, T.; Zhu, L.; Ding, H.; Sheng, Z.; Welihinda,
A.; Roberge, J. Y.; Seed, B.; Chen, Y. Bioorg. Med. Chem. Lett. 2011, 21, 4465.
22. Tahara, A.; Matsuyama-Yokono, A.; Nakano, R.; Someya, Y.; Shibasaki, M. Basic
Clin. Pharmacol. Toxicol. 2008, 103, 560.
5.2.4. Urinary glucose excretion in rats
Male Wistar rats, aged 8 weeks, were housed in metabolic
cages. Rats were fasted from morning (08:00 hours) to the next
morning. In the afternoon (16:00 hours), rats were administered
test compounds (10 mg/kg) suspended in 0.5% CMC or vehicle or-
ally. Urine was then collected until the following morning (08:00
hours). After measuring urine volume, an aliquot of each urine
sample was stored at ꢁ20 °C until use. The glucose concentration
in urine was measured using the glucose oxidase method. An F-test
followed by Student’s t-test or Welch’s test was used to estimate
the degree of statistical significance of the difference between
the vehicle group and the test compound group.
23. (a) Lamanna, C.; Bellini, M.; Padova, A.; Westerberg, G.; Maccari, L. J. Med.
Chem. 2008, 51, 2891; (b) Ritchie, T. J.; Macdonald, S. J. F. Drug Discovery Today
2009, 14, 1011.
24. Kees, K. L.; Caggiano, T. J.; Steiner, K. E.; Fitzgerald, J. J.; Kates, M. J.; Christos, T.
E.; Kulishoff, J. M.; Moore, R. D.; McCaleb, M. L. J. Med. Chem. 1995, 38, 617.
25. Ikenoue, T.; Okazaki, K.; Fujitani, S.; Tsuchiya, Y.; Akiyoshi, M.; Maki, T.; Kondo,
N. Biol. Pharm. Bull. 1997, 20, 354.
Acknowledgments
The authors acknowledge Dr. Eiichi Tsuji, Dr. Hideyuki
Muranaka and Dr. Yoshinori Nonaka for X-ray crystal analysis,
HRMS and NMR measurements of the compounds, respectively.
We also thank Dr. Robert L. Dobbins of GlaxoSmithKline, Dr. Har-