5a-Carba-β-d-glucopyranose derivatives as novel sodium-dependent glucose cotransporter 2 (SGLT2) inhibitors for the treatment of type 2 diabetes

Article abstract of DOI:10.1016/j.bmc.2011.08.005

5a-Carba-β-d-glucopyranose derivatives were synthesized and identified as novel SGLT2-selective inhibitors. These inhibitors exhibited potent SGLT2 inhibition with high selectivity over SGLT1. Among the tested compounds, 6f indicated the most potent hSGLT2 inhibition and the highest selectivity over hSGLT1. Moreover, the pharmacokinetics data also showed that 6h, which had the same aglycon structure as sergliflozin-active (3-active), had a threefold longer half-life time (T_1/2) than sergliflozin (3) with a high distribution volume in db/db mice. Subsequently, 6h lowered blood glucose levels as much as 3 and showed longer hypoglycemic action than 3 in db/db mice.

Full text of DOI:10.1016/j.bmc.2011.08.005

Bioorganic & Medicinal Chemistry 19 (2011) 5334–5341
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
5a-Carba-b-D-glucopyranose derivatives as novel sodium-dependent glucose
cotransporter 2 (SGLT2) inhibitors for the treatment of type 2 diabetes
⇑
Yoshihito Ohtake , Tsutomu Sato, Hiroharu Matsuoka, Masahiro Nishimoto, Naoki Taka, Koji Takano,
Keisuke Yamamoto, Masayuki Ohmori, Takashi Higuchi, Masatoshi Murakata, Takamitsu Kobayashi,
Kazumi Morikawa, Nobuo Shimma, Masayuki Suzuki, Hitoshi Hagita, Kazuharu Ozawa, Koji Yamaguchi,
Motohiro Kato, Sachiya Ikeda
Research Division, Fuji Gotemba Research Laboratory, Chugai Pharmaceutical Co., Ltd, 1-135, Komakado, Gotemba, Shizuoka 412-8513, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
5a-Carba-b-D-glucopyranose derivatives were synthesized and identified as novel SGLT2-selective inhib-
Received 16 June 2011
Revised 2 August 2011
Accepted 3 August 2011
Available online 8 August 2011
itors. These inhibitors exhibited potent SGLT2 inhibition with high selectivity over SGLT1. Among the
tested compounds, 6f indicated the most potent hSGLT2 inhibition and the highest selectivity over
hSGLT1. Moreover, the pharmacokinetics data also showed that 6h, which had the same aglycon struc-
ture as sergliflozin-active (3-active), had a threefold longer half-life time (T1/2) than sergliflozin (3) with
a high distribution volume in db/db mice. Subsequently, 6h lowered blood glucose levels as much as 3
and showed longer hypoglycemic action than 3 in db/db mice.
Keywords:
Type 2 diabetes
SGLT2 inhibitor
Carbasugar
Ó 2011 Elsevier Ltd. All rights reserved.
5a-Carba-b-D-glucopyranose
Metabolic stabilization
1. Introduction
O-glycosidic type of SGLT inhibitor, and repeated subcutaneous
administration of 1 lowered the blood glucose levels of diabetic
Type 2 diabetes (T2D) mellitus is a progressive metabolic disease
characterized by decreased insulin secretion and increased insulin
resistance, the prevalence of which has been increasing yearly.
However, it is generally accepted that current medication does not
adequately control the blood glucose in patients with diabetes. In
addition, hypoglycemia is reported as one of the side effects when
insulin or sulfonylureas is administered. Therefore, more effective
and safer new drugs for the treatment of T2D are strongly desired.
In the last decade, sodium glucose co-transporter 2 (SGLT2) has
been attracting attention because it is uniquely responsible for
reabsorbing glucose from the renal filtrate.1 The expression site
of SGLT2 is primarily the proximal convoluted tubules, whereas
those of SGLT1 are small intestine, heart, brain, and renal tubules.
Furthermore, SGLT1 mutation is known to cause glucose-galactose
malabsorption,2 so SGLT1 inhibition is reported to possibly cause
gastrointestinal side effects.3 From these findings, SGLT2-selective
inhibitors are considered to be safer than non-selective inhibitors.
The reported SGLT2 inhibitors are classified into O-aryl gluco-
pyranosides and C-aryl glucopyranosides by the type of glycosidic
bond (Fig. 1). The natural product phlorizin (1) is a non-selective
rodents.4 T-1095 (2), which was reported by the Tanabe group,
was the first time a structural modification of phlorizin was used
to discover a selective SGLT2 inhibitor.5 However, this inhibitor
was considered to be metabolically unstable because of the cleav-
age of the O-glycosidic bond by glycosidases, so a methyl carbonate
prodrug was required to avoid high dosing. Subsequently,
several groups have reported potent SGLT2-selective inhibitors of
O-glycosides, such as sergliflozin (3)6 and remogliflozin etabonate,7
but these compounds have not been launched yet. On the other
hand, the Bristol–Myers Squibb group has recently disclosed the
discovery of a highly potent SGLT2 inhibitor, dapagliflozin (4),8
which has the benefit of long-lasting action due to being a meta-
bolically stable C-aryl glycosidic type. Likewise, canagliflozin (5)9
has been reported by the Mitsubishi Tanabe group. Thus, O-type
inhibitors are generally considered to be metabolically more unsta-
ble than C-type inhibitors making it difficult to achieve a lower
dose in vivo, however, we decided to strive for the discovery of
novel O-type SGLT2 inhibitors which are metabolically stable and
effective in diabetic animal models.
One of our strategies for discovering metabolically stable novel
SGLT2 inhibitors was to convert unstable O-glycoside to stable
glucose mimics, effectively avoiding the cleavage of the O-glyco-
sidic bond by glycosidases (Fig. 2). Some investigations of the
structural modification of sugar analog have been reported as
⇑
Corresponding author.
E-mail address: ohtakeysh@chugai-pharm.co.jp (Y. Ohtake).
0968-0896/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2011.08.005

Y. Ohtake et al. / Bioorg. Med. Chem. 19 (2011) 5334–5341
5335
OH
O
O
OH
O
O
O
OMe
O
Me
HO
OH
OH
OH
OH
OH
OH
OH
O
O
O
O
MeO
O
HO
R
OH
O
OH
OH
3 sergliflozin (R=CO2Et)
1 phlorizin
2 T-1095
(O-glucopyranoside)
(O-glucopyranoside)
3-active (R=H)
(O-glucopyranoside)
Me
Cl
S
F
O
OH
O
OH
OH
O
HO
HO
OH
OH
OH
4 dapagliflozin
(C-glucopyranoside)
5 canagliflozin
(C-glucopyranoside)
Figure 1. Structures of some known SGLT inhibitors.
OH
OBn
OBn
R
Several steps
R
Sugar part modification
O
2 steps
O
O
HO
BnO
BnO
OH
OBn
OH
OH
OH
OH
O
OH
OBn
OBn
HO
metabolic stabilization
Duration of action
HO
7 (D-glucal)
8
9
OH
6a-6n
OH
Scheme 1. Synthesis of 2,3,4,6-tetra-O-benzyl-5a-carba-
a-
D-glucopyranose (9)
(O-glucopyranoside type)
(O-carba-glucopyranoside type)
Figure 2. Our original design of carba-glucopyranoside SGLT2 inhibitors.
11e; R=iPr
11f; R=cPr
11i; R=OEt
11j; R=SMe
11l; R=F
SGLT2-selective inhibitors,10 but there were no reports of an SGLT2
inhibitor bearing a carbasugar.11 Although examples of carbasugar
being reported as a glucose mimetic were limited, those examples
have encouraged us to prepare and evaluate carbasugar derivatives
as SGLT2 inhibitors (6a–6n).12
(Method A)a, b
Br
OBn
OH
R
(Method B)a, c, d
10
11m; R=Cl
11n; R=CF
3
2. Results and discussion
2.1. Chemistry
Scheme 2. Reagents and conditions: (a) nBuLi, THF, À78 °C then 4-substituted
benzaldehyde; (b) Pd(OH)2, H2, 2 N HCl aq, MeOH/AcOEt, rt; (c) Et3SiH, BF3ÁOEt2,
MeCN, À40 °C; (d) Me2S, BF3ÁOEt2, CH2Cl2, 0 °C to rt (11e; 73%, 11f; 56%, 11i; 38%,
11j; 50%, 11l; 77%, 11m; 62%, 11n; 71%).
First, we prepared 2,3,4,6-tetra-O-benzyl-5a-carba-
a-
D
-gluco-
pyranose (9) as a key intermediate from
D-glucal (7) by using Oga-
wa’s method13 and Sudha’s method14 (Scheme 1).
O
R
Next, the general synthetic methods of aglycons 11e–11f,
11i–11j, 11l–11n are shown in Scheme 2. After lithium halogen
exchange of the commercially available 2-benzyloxybromoben-
zene (10), 4-substituted benzaldehyde derivatives were added to
give the corresponding alcohols in good yield. The alcohol deriva-
tives were converted to 11 by direct route (Method A) or stepwise
route (Method B), depending on their reactivity of R substituent in
reduction conditions.
The general synthesis of the final carbasugar derivatives is
shown in Scheme 3. Our key reaction was the coupling reaction be-
tween carbasugar 9 and the corresponding aglycon 11 with inver-
sion of C-1 position on the cyclohexyl ring. In our investigation, the
coupled compounds were obtained by the combination of tri-
nbutylphosphine (nBu3P) and tetramethylazodicarbox-amide
(TMAD). Finally, removal of the tetra benzyl groups furnished the
desired carba-glucopyranose compounds 6a–6f, 6i–6j, 6l–6n.
The compound 6g (R = CH@CH2) was prepared as shown in
Scheme 4. Key intermediate 9 was treated with trifluoromethane-
sulfonic anhydride, and coupled with sodium 2-(4-bromophe-
nylmethyl)phenoxide 1315 with inversion. After deprotection of
a
b
OH
OH
9
HO
OH
OH
R
6a-6f, 6i-6j, 6l-6n
11
Scheme 3. Reagents and conditions: (a) compound 11, nBu3P, TMAD, toluene/THF,
0 °C to rt; (b) Pd(OH)2, H2, MeOH/THF, rt or Me2S, BF3ÁOEt2, CH2Cl2, 0 °C to rt (6a;
26%, 6b; 6%, 6c; 14%, 6d; 21%, 6e; 27%, 6f; 25%, 6i; 36%, 6j; 28%, 6l; 46%, 6m; 35%,
6n; 19%).
four benzyl groups, vinylation was finally implemented by palla-
dium coupling reaction with vinyl tributylstannane to afford the
vinyl compound 6g.
As shown in Scheme 5, 6h (R = OMe) and 6k (R = OH) were also
synthesized. Coupling reaction of 9 and methyl salicylate was con-
ducted under Mitsunobu condition to afford 15 with the inversion
of C-1 position on carbasugar. After reducing the methyl ester,
benzaldehyde 17 was obtained in good yield by using Dess–Martin
reagent. It was treated with commercially available 4-methoxy-
phenyl Grignard reagent to afford compound 18, followed by

5336
Y. Ohtake et al. / Bioorg. Med. Chem. 19 (2011) 5334–5341
Table 1
OTf
c
b
In vitro data for hSGLT inhibition and selectivity
OBn
OBn
a
9
BnO
OBn
OH
Br
12
O
R
13
OH
OH
HO
OH
O
Br
O
d
Selectivityb
OH
OH
OH
OH
Compds
R
IC50 (nM)a
HO
HO
hSGLT2
hSGLT1
OH
OH
1
3
16
260
190
22,000
2,300
13,000
6,700
4,800
7,800
20,000
5,600
3,300
9,300
41,000
3,500
19,000
38,000
22,000
57,000
12
85
480
19
14
6g
3-Active
6a
6b
6c
6d
6e
6f
6g
6h
6i
6j
6k
6l
6m
6n
4.8
H
Me
Et
nPr
iPr
cPr
Vinyl
OMe
OEt
SMe
OH
F
690
59
28
120
150
11
Scheme 4. Reagents and conditions: (a) Tf2O, pyridine, CH2Cl2, 0 °C; (b) 60% NaH,
DMF, À40 °C to 0 °C; (c) Me2S, BF3ÁOEt2, CH2Cl2, 0 °C to rt (43% for 3 steps); (d)
nBu3SnCH@CH2, Pd(PPh3)4, 2,6-di-tert-Bu-4-MeC6H3OH, toluene, 110 °C (26%).
110
170
65
130
510
110
220
370
180
66
CO2Me
29
42
OH
O
110
19
290
630
150
120
OBn
a
OBn
OBn
BnO
OBn
CO2Me
OH
BnO
60
150
480
OBn
OBn
Cl
CF3
9
15
a
The IC50 values are means of multiple independent experiments. Compound 1
CHO
O
OH
and 3-active as reference standards were always included in the assays.
b
O
The selectivity values were calculated by IC50 hSGLT1/IC50 hSGLT2.
b
c
OBn
OBn
OBn
OBn
BnO
BnO
selectivity. Moreover, the branched and cyclic alkyl substituents
were also investigated (6e, 6f). The most noteworthy compound
was the cyclopropyl-substituted compound 6f, which had the most
potent hSGLT2 inhibitory activity (11 nM) and the highest selectiv-
ity against hSGLT1 (510-fold). The vinyl-substituted compound
(6g) exhibited the same potency as 6c. The methoxy substitution
(6h), which is the same substituent as sergliflozin (3), was found
to exhibit potent inhibition of hSGLT2. When the methoxy group
was replaced with the methylthio group (6j), the potency of
hSGLT2 inhibition was doubled. Substitution at this position with
the hydrophilic hydroxyl group (6k) resulted in a loss of potency.
The introduction of a small sized fluoro group (6l) showed the
same result as 6a. The other halogen derivatives (6m–6n) also gave
moderate inhibitory activity.
As a consequence of our investigation, we demonstrated that
hSGLT1 and hSGLT2 inhibitions were affected by the substituent R.
Interestingly, the order of hSGLT2 IC50 values in alkyl substituent
series (6a–6g, including the vinyl group) is as follows: 6a
(R = H) >> 6b (R = Me) > 6c (R = Et) = 6g (R = vinyl) > 6f (R = cPr) <
6d (R = nPr) < 6e (R = iPr). These results suggest that the substituent
at para position on the distal benzene ring occupies a two- or three-
carbon-sized pocket of hSGLT2. On the other hand, inhibitors bear-
ing a relatively hydrophilic substituent such as 6h (R = OMe), 6i
(R = OEt), and 6k (R = OH) exhibited less potent inhibition, com-
pared with inhibitors which have a correspondingly sized alkyl
group. These results suggest that the pocket of hSGLT2 is hydropho-
bic. Three halogen substituted compounds (6l–6n), which are smal-
ler in size than a cyclopropyl substituent, also showed relatively
weak inhibition, suggesting that these inhibitors were not satisfac-
tory either in size or hydrophobicity. Meanwhile, these compounds
inhibited hSGLT1 considerably more weakly than they did hSGLT2.
Importantly, all the tested compounds showed hSGLT2 selectivity
over hSGLT1, extensively. From these findings, having a substituent
of suitable size and hydrophobicity at para position on the distal
benzene ring is considered to be critical for potent hSGLT2 inhibition
and higher selectivity than to hSGLT1.
OBn
16
OBn
17
OH
OMe
R
O
O
d
e
OBn
OBn
OH
OH
BnO
HO
OBn
OH
18
6h; R=OMe
6k; R=OH
f
Scheme 5. Reagents and conditions: (a) PPh3, DEAD, THF, rt (49%); (b) LiAlH4, THF,
55 °C (81%); (c) Dess–Martin reagent, CH2Cl2, rt (81%); (d) 0.5 M 4-MeOC6H4MgBr in
THF solution, Et2O, rt (74%); (e) Pd(OH)2, H2, HCl–MeOH, rt (61%); (f) BBr3, CH2Cl2,
À78 °C to rt (47%).
dehydroxylation at 1,1-diphenylmethy position and deprotection
of four benzyl groups, which were conducted simultaneously to
give the methoxy compound 6h. Finally, demethylation of the
methoxy group on the distal benzene ring was conducted by use
of boron tribromide (BBr3) and gave 6k in moderate yield.
2.2. Results and structure–activity relationship
We evaluated all the synthesized compounds 6a–6n for in vitro
inhibitory activity against human SGLT1 (hSGLT1) and human
SGLT2 (hSGLT2) in order to investigate the effect of substituent
(R) at para position on the distal benzene ring. The results are
shown in Table 1. Compound 6a (R = H) had a weak inhibitory
activity for hSGLT2. Introduction of a methyl group (6b) resulted
in the enhancement of hSGLT2 inhibition and selectivity against
hSGLT1 compared with 6a. By replacing the methyl group with
an ethyl group (6c), its hSGLT2 inhibition and selectivity over
hSGLT1 was further improved, whereas the introduction of a pro-
pyl substituent (6d) resulted in weak hSGLT2 inhibition and low

Y. Ohtake et al. / Bioorg. Med. Chem. 19 (2011) 5334–5341
5337
Table 2
Pharmacokinetic parameters of db/db mice for key compounds
4. Experimental
Compds
Dose
(mg/kg; po)
Cmax
g/mL)
CL/F
(mL/h/kg)
Vz/Fa
(mL/kg)
T1/2
(h)
4.1. Chemistry: instruments
(
l
Silica gel 60F254 precoated plates on glass from Merck KgaA
were used for thin layer chromatography (TLC). Column chroma-
tography was carried out on Merck Silica Gel 60 (230–400 mesh),
if not otherwise specified. Reverse phase column chromatography
was carried out on Fuji Silysia Chromatorex Pro. No. DM2035MT.
1H NMR spectra were recorded on JEOL EX-270 (270 MHz), Varian
Mercury300 (300 MHz) or JEOL JNM-ECP-400 (400 MHz), and
chemical shifts were reported in parts per million (d) downfield
from tetramethylsilane as an internal standard. The peak patterns
are shown as the following abbreviations: br, broad; s, singlet; d,
doublet; t, triplet; q, quartet; and m, multiplet. Mass spectra
(MS) were measured by a Thermo Electron LCQ Classic or Micro-
mass ZQ of Waters (ESI). High resolution mass spectra (HRMS)
were recorded by a Micromass Q-Tof Ultima API mass spectrome-
ter. All reagents and the solvent were commercially available un-
less otherwise indicated.
3
6h
10
10
3.36
1.64
2,818
3,257
3,315
12,586
0.82
2.68
a
The apparent volume of distribution during terminal phase at oral
administration.
4.1.1. 2,3,4,6-Tetra-O-benzyl-5a-carba-
2,3,4,6-Tetra-O-benzyl-5a-carba- -glucopyranose (9) was
-glucal (7) in several
a-D-glucopyranose (9)
a-D
synthesized from commercially available
D
steps.
Figure 3. Blood glucose lowering effect of single oral administration of 3 and 6h in
db/db mice.
4.1.2. Aglycons (11a, 11b, 11c)
2.3. PK and in vivo studies
Aglycons 11a and 11b were commercially available. These were
used without further purification. 11c was prepared by the proce-
dure of the patent.16
Among the carbasugar derivatives, 6h was chosen for pharma-
cokinetic study in db/db mice to compare with 3 because 6h had
the same aglycon structure as 3-active and the difference between
compounds was only one atom structurally (Table 2). By removing
the glycosidic bond, which is cleavable by b-glycosidase, we ex-
pected that the PK profile, especially the time of half-life (T1/2),
would be prolonged. In the results, 6h showed threefold longer
T1/2 than 3, as expected. Even though the cleavage of the glycosidic
bond by b-glycosidase was successfully avoided (data not shown),
the clearance (CL/F) of 6h was not improved. The poor clearance
might be attributed to metabolic factors other than cleavage of
the glycosidic bond. As for the reason for the longer T1/2, it resulted
from the distribution volume (Vz/F) being greatly elevated. That is
to say, the structural modification of glucose analog to carbasugar
analog might increase the compound’s lipophilicity and enhance
distribution in tissue.
Furthermore, we studied the blood glucose lowering effect in
db/db mice (Fig. 3). The resulting effects of 6h (100 mg/kg) were
as strong as those of 3 (100 mg/kg) even though each hSGLT2 inhi-
bition value varied greatly (IC50: 4.8 nM vs 42 nM). Moreover, 8 h
after dosing, the blood glucose levels in the 6h-treated mice were
lower than those in the 3-treated mice. This means that, as we ex-
pected from the PK profiles, 6h has a longer-lasting effect than 3.
4.1.3. 2-(4-Isopropylbenzyl)phenol(11e) (direct route, method A)
In
a nitrogen stream, 2.44 M nBuLi in hexane (5.14 mL,
12.5 mmol) was added dropwise to a solution of 1-benzyloxy-2-
bromobenzene (3.0 g, 11.4 mmol) in THF (114 mL) at À78 °C and
the mixture was stirred at the same temperature for 20 min. To
this solution,
a solution of 4-isopropylbenzaldehyde (1.41 g,
9.49 mmol) in THF (38 mL) was added dropwise at À78 °C. The
mixture was stirred at the same temperature for 2 h, and then sat-
urated aq NH4Cl was added thereto and the mixture was extracted
with AcOEt. The organic layer was washed with brine and dried
with MgSO4, and then the solvent was removed under reduced
pressure. The obtained residue was purified by silica gel column
chromatography (AcOEt/hexane = 1:5) to obtain 2-benzyloxyphe-
nyl-4-isopropylphenylmethanol (2.63 g, 84%). To a solution of the
obtained product in MeOH (50 mL), 20% Pd(OH)2 (263 mg) was
added and furthermore 2 N HCl (400 lL) was added thereto. The
mixture was stirred under a hydrogen atmosphere for 15 h, and
then the catalyst was filtered. The solvent was removed under re-
duced pressure and the obtained residue was purified by silica gel
column chromatography (AcOEt/hexane = 1:4) to obtain the title
compound (1.31 g, 73%). 1H NMR (CDCl3) d: 1.24 (6H, d,
J = 7.8 Hz), 2.82–2.92 (1H, m), 3.96 (2H, s), 4.67 (1H, s), 6.79 (1H,
d, J = 9.0 Hz), 6.88 (1H, t, J = 8.4 Hz), 7.10–7.15 (6H, m).
3. Conclusion
4.1.4. 2-(4-Cyclopropylbenzyl)phenol (11f) (stepwise route,
method B)
We introduced the metabolically stable O-Aryl 5a-carba-b-D-
glucopyranosides 6a–6n as novel SGLT2 inhibitors with a carba-
sugar analog and revealed the SAR for the substituent on the distal
benzene ring. Among the tested compounds, 6f (R = cPr) indicated
the most potent hSGLT2 inhibition and the highest selectivity over
hSGLT1. The pharmacokinetics data also showed that 6h (R = OMe)
had a three-fold longer T1/2 than sergliflozin (3) with a high distri-
bution volume in db/db mice. Subsequently, 6h lowered blood glu-
cose levels as much as 3 and showed longer hypoglycemic action
than 3 in db/db mice. Further investigations of SGLT2 inhibitors
having a carbasugar analog will be reported in the near future.
4.1.4.1. 2-Benzyloxyphenyl-(4-cyclopropylphenyl)methanol. In
a nitrogen stream, 2.6 M nBuLi in hexane (3.5 mL, 9.1 mmol) was
added dropwise to a solution of 1-benzyloxy-2-bromobenzene
(2.2 g, 8.3 mmol) in THF (83 mL) at À78 °C and the mixture was
stirred at the same temperature for 30 min. To this solution, a solu-
tion of 4-cyclopropylbenzaldehyde (1.1 g, 6.9 mmol) in THF
(280 mL) was added dropwise at À78 °C. The reaction mixture
was stirred at the same temperature for 1 h. Water was added
thereto and the mixture was extracted with AcOEt. The organic

5338
Y. Ohtake et al. / Bioorg. Med. Chem. 19 (2011) 5334–5341
layer was washed with brine and dried with MgSO4, and then the
solvent was removed under reduced pressure. The obtained
residue was purified by silica gel column chromatography
(AcOEt/hexane = 1:5) to obtain the title compound (1.7 g, 75%).
1H NMR (CDCl3) d: 0.65–0.69 (2H, m), 0.92–0.97 (2H, m), 1.86–
1.90 (1H, m), 2.88 (1H, d, J = 6.0 Hz), 5.03 (2H, s), 6.03 (1H, d,
J = 6.0 Hz), 7.17–7.26 (5H, m), 7.32–7.35 (4H, m); MS (ESI) m/z:
315 ([M+Na]+).
4.1.9. 2-(4-Trifluoromethylbenzyl)phenol (11n)
This compound was prepared from 1-benzyloxy-2-bromoben-
zene and 4-trifluoromethylbenzaldehyde in the same manner as
described in Section 4.1.3. Yield 71%. 1H NMR (CDCl3) d: 4.03
(2H, s), 4.72 (1H, s), 6.77 (1H, d, J = 6.0 Hz), 6.89 (1H, t,
J = 4.8 Hz), 7.08–7.15 (2H, m), 7.33 (2H, d, J = 6.0 Hz), 7.52 (2H, d,
J = 6.0 Hz); MS (ESI) m/z: 275 ([M+Na]+).
4.1.10. (2-Benzylphenyl)-5a-carba-b-
D
-glucopyranoside (6a)
-glu-
4.1.4.2. 1-Benzyloxy-2-(4-cyclopropylbenzyl)benzene. In a nitro-
gen stream, Et3SiH (0.73 mL, 4.6 mmol) and BF3ÁEt2O (0.5 mL,
4.0 mmol) were added to a solution of 2-benzyloxyphenyl-4-cyclo-
propylphenyl-methanol (1.3 g, 4.0 mmol) in MeCN (7 mL) at À40 °C
and the mixture solution was stirred at the same temperature for
1.5 h and furthermore at 0 °C for 30 min. Water was added thereto
and the mixture was extracted with CH2Cl2. The organic layer was
washed with brine and dried with MgSO4, and then the solvent
was removed under reduced pressure. The obtained residue was
purified by silica gel column chromatography (AcOEt/hexane = 1:7)
to obtain the title compound (1.2 g, 95%). 1H NMR (CDCl3) d:
0.63–0.67 (2H, m), 0.89–0.94 (2H, m), 1.84–1.87 (1H, m), 3.98
(2H, s), 5.06 (2H, s), 6.87–6.97 (4H, m), 7.08–7.26 (4H, m),
7.31–7.37 (5H, m); MS (ESI) m/z: 332 ([M+H2O]+).
In a nitrogen stream, 2,3,4,6-tetra-O-benzyl-5a-carba-
a-D
copyranose (9) (500 mg, 0.93 mmol) and nBu3P (0.35 mL,
1.39 mmol) were added to a solution of commercially available
2-benzylphenol (257 mg, 1.39 mmol) in toluene (2 mL) under cool-
ing with ice, and then tetramethylazodicarboxamide (TMAD,
239 mg, 1.39 mmol) was added thereto at the same temperature.
The reaction mixture was stirred for 21 h while gradually raising
its temperature to room temperature. The reaction mixture was
concentrated under reduced pressure, and the obtained residue
was treated by silica gel column chromatography (AcOEt/hex-
ane = 1:10). The obtained crude product was dissolved in
a
MeOH/THF (1:1) mixed solution (14 mL) and 20% Pd(OH)2
(119 mg) was added thereto and the mixture was stirred under a
hydrogen atmosphere for 15 h, and then the catalyst was filtered
off. The solvent was removed under reduced pressure and the ob-
tained residue was purified by silica gel column chromatography
(MeOH/CH2Cl2 = 1:10) to obtain the title compound (84 mg, 26%)
as a colorless amorphous solid. 1H NMR (CD3OD) d: 0.95 (1H, dd,
J = 11.7, 12.0 Hz), 1.49–1.59 (1H, m), 2.03–2.09 (1H, m), 3.20 (1H,
d, J = 9.0 Hz), 3.25 (1H, d, J = 5.7 Hz), 3.44–3.50 (2H, m), 3.68 (1H,
dd, J = 3.9, 4.2 Hz), 3.94 (1H, d, J = 15.0 Hz), 4.00 (1H, d,
J = 15.0 Hz), 4.13–4.22 (1H, m), 6.84 (1H, t, J = 6.3 Hz), 7.03 (1H,
d, J = 6.3 Hz), 7.06–7.24 (7H, m); MS (ESI) m/z: 345 ([M+H]+); HRMS
calcd for C20H23O5 ([MÀH]À) 343.154. Found 343.1539.
4.1.4.3. 2-(4-cyclopropylbenzyl)phenol. In
a nitrogen stream,
Me2S (2.2 mL, 51.7 mmol) and BF3ÁEt2O (1.1 mL, 8.8 mmol) were
added to a solution of 1-benzyloxy-2-(4-cyclopropyl-benzyl)ben-
zene (1.1 g, 3.5 mmol) in CH2Cl2 (24 mL) under cooling with ice,
and the reaction mixture was stirred for 23 h while gradually rais-
ing its temperature to room temperature. Water was added thereto
under cooling with ice and the mixture was extracted with CH2Cl2.
The organic layer was washed with brine and dried with MgSO4,
and then the solvent was removed under reduced pressure. The ob-
tained residue was purified by silica gel column chromatography
(AcOEt/hexane = 1:4) to obtain the title compound (621 mg, 79%).
1H NMR (CDCl3) d: 0.65–0.67 (2H, m), 0.90–0.94 (2H, m), 1.85–
1.86 (1H, m), 3.95 (2H, s), 4.90 (1H, s), 6.79 (1H, d, J = 8.1 Hz),
6.88 (1H, t, J = 7.7 Hz), 7.11 (2H, d, J = 8.1 Hz), 7.20 (4H, d,
J = 7.7 Hz); MS (ESI) m/z: 247 ([M+Na]+).
4.1.11. [2-(4-Methylbenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6b)
This compound was prepared from 9 and 2-(4-methylben-
zyl)phenol (11b) in the same manner as described in Section
4.1.10. Yield 6%. 1H NMR (CD3OD) d: 0.84–0.97 (1H, m), 1.44–2.03
(1H, m), 1.96–2.03 (1H, m), 2.22 (3H, s), 3.13–3.30 (2H, m),
3.39–3.46 (2H, m), 3.64 (1H, dd, J = 4.5, 10.8 Hz), 3.95 (2H, m),
4.12 (1H, m), 6.76–6.81 (1H, m), 6.92–7.03 (5H, m), 7.06–7.11
(2H, m); MS (ESI) m/z: 381 ([M+Na]+); HRMS calcd for C21H25O5
([MÀH]À) 357.1697. Found 357.1694.
4.1.5. 2-(4-Ethyoxybenzyl)phenol (11i)
This compound was prepared from 1-benzyloxy-2-bromoben-
zene and 4-ethoxybenzaldehyde in the same manner as described
in Section 4.1.3. Yield 38%. 1H NMR (CDCl3) d: 1.39 (3H, t,
J = 7.0 Hz), 3.93 (2H, s), 3.99 (2H, q, J = 7.0 Hz), 6.77–6.90 (4H, m),
7.09–7.14 (4H, m).
4.1.12. [2-(4-Ethylbenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6c)
4.1.6. 2-(4-Methylsulfanylbenzyl)phenol (11j)
This compound was prepared from 9 and 2-(4-ethylbenzyl)phe-
nol (11c) in the same manner as described in Section 4.1.10. Yield
14%. 1H NMR (CD3OD) d: 0.86–1.00 (1H, m), 1.19 (3H, t, J = 7.6 Hz),
1.45–1.62 (1H, m), 2.00–2.11 (1H, m), 2.57 (2H, q, J = 7.6 Hz), 3.16–
3.30 (2H, m), 3.43–3.49 (2H, m), 3.70 (1H, dd, J = 4.1, 10.6 Hz),
3.84–4.02 (2H, m), 4.12–4.24 (1H, m), 6.83 (1H, t, J = 7.4 Hz),
6.99–7.16 (7H, m); MS (ESI): 395 ([M+Na]+); HRMS calcd for
This compound was prepared from 1-benzyloxy-2-bromoben-
zene and 4-methylsulfanylbenzaldehyde in the same manner as
described in Section 4.1.4. Yield 50%. 1H NMR (CDCl3) d: 2.46
(3H, s), 3.95 (2H, s), 4.65–4.75 (1H, br s), 6.78 (1H, d, J = 7.6 Hz),
6.89 (1H, t, J = 7.6 Hz), 7.09–7.21 (6H, m).
4.1.7. 2-(4-Fluorobenzyl)phenol (11l)
C
22H27O5 ([MÀH]À) 371.1853. Found 371.1850.
This compound was prepared from 1-benzyloxy-2-bromoben-
zene and 4-fluorobenzaldehyde in the same manner as described
in Section 4.1.3. Yield 77%. 1H NMR (CDCl3) d: 3.95 (2H, s), 4.72
(1H, s), 7.77 (1H, dd, J = 1.1, 7.9 Hz), 6.88 (1H, dt, J = 1.1, 7.5 Hz),
6.96 (1H, dd, J = 8.7, 8.7 Hz), 7.06–7.20 (4H, m).
4.1.13. [2-(4-nPropylbenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6d)
1-Benzyloxy-2-(4-cyclopropylbenzyl)benzene (641 mg, 2.0 mmol)
as synthesized in Section 4.1.4. was dissolved in
4.1.8. 2-(4-Chlorobenzyl)phenol (11m)
2,2-dimethylpropanol (12 mL). 20% Pd(OH)2 (64 mg) was added
thereto and the mixture was stirred for 2 h under a hydrogen
atmosphere, and then the catalyst was filtered off. The solvent was
removed under reduced pressure, and the obtained residue was
treated by silica gel column chromatography (AcOEt/hexane = 1:4).
To a solution of the obtained 2-(4-nPropylbenzyl)phenol (348 mg)
This compound was prepared from 1-benzyloxy-2-bromoben-
zene and 4-chlorobenzaldehyde in the same manner as described
in Section 4.1.4. Yield 62%. 1H NMR (CDCl3) d: 3.95 (2H, s),
4.60 (1H, s), 6.75–6.78 (1H, m), 6.86–6.92 (1H, m), 7.07–7.27
(6H, m).

Y. Ohtake et al. / Bioorg. Med. Chem. 19 (2011) 5334–5341
5339
in toluene (3.5 mL), 9 (557 mg, 1.03 mmol) and nBu3P (0.39 mL,
1.55 mmol) were added under cooling with ice, and then TMAD
(267 mg, 1.55 mmol) was added thereto at the same temperature.
The reaction mixture was stirred for 20 h while gradually raising
its temperatureto roomtemperature. The reaction mixturewas con-
centrated under reduced pressure, and the obtained residue was
treated with silica gel column chromatography (AcOEt/hex-
ane = 1:5). The obtained product was dissolved in 2,2-dimethylpro-
panol (15 mL). 20% Pd(OH)2 (159 mg) was added thereto and the
mixture was stirred under a hydrogen atmosphere for 17 h, and
the catalyst was filtered off. The solvent was removed under reduced
pressure, and the obtained residue was purified by silica gel column
chromatography (MeOH/CH2Cl2 = 1:9) to obtain the title compound
(165 mg, 21%) as a colorless amorphous solid. 1H NMR (CD3OD) d:
0.88–0.94 (4H, m), 1.48–1.63 (3H, m), 2.05–2.09 (1H, m), 2.52 (2H,
t, J = 7.7 Hz), 3.13–3.30 (2H, m), 3.46 (2H, m), 3.70 (1H, m), 3.87
(1H, d, J = 15.0 Hz), 3.98 (1H, d, J = 15.0 Hz), 4.60–4.88 (1H, m),
6.84 (1H, t, J = 7.3 Hz), 7.00–7.16 (7H, m); MS (ESI) m/z: 387
([M+H]+); HRMS calcd for C23H29O5 ([MÀH]À) 385.2010. Found
385.2011.
(205
0.557 mmol) in CH2Cl2 (5.5 mL) and the mixture was cooled to
0 °C, and then Tf2O (210 L, 1.25 mmol) was added thereto. The
lL, 2.53 mmol) was added to a solution of 9 (300 mg,
l
reaction mixture was stirred at the same temperature for 1 h,
and then saturated aq NaHCO3 was added thereto and the mixture
was extracted with CH2Cl2. The combined organic layer was
washed with saturated aq KHSO4 and brine, and dried over MgSO4.
The solvent was removed to obtain the crude titled product
(380 mg). 1H NMR (CDCl3) d: 1.83 (1H, dd, J = 14.0, 15.9 Hz),
2.00–2.14 (2H, m), 3.39 (1H, dd, J = 2.3, 9.2 Hz), 3.50 (1H, dd,
J = 2.6, 9.6 Hz), 3.56 (1H, dd, J = 9.3, 10.2 Hz), 3.75 (1H, dd, J = 3.4,
9.2 Hz), 3.86 (1H, dd, J = 9.3, 9.5 Hz), 4.40 (2H, s), 4.50 (1H, d,
J = 10.8 Hz), 4.62 (1H, d, J = 11.4 Hz), 4.79 (1H, d, J = 10.7 Hz), 4.82
(1H, d, J = 11.5 Hz), 4.89 (1H, d, J = 10.7 Hz), 4.92 (1H, d,
J = 10.7 Hz), 5.33 (1H, br), 7.15–7.30 (20H, m).
4.1.16.2. 2-(4-Bromobenzyl)phenol (13). In a nitrogen stream,
Me2S (1.83 mL, 42.46 mmol) and BF3ÁEt2O (0.9 mL, 7.07 mmol)
were added to a solution of 1-benzlyoxy-2-(4-bromobenzyl)ben-
zene (1.0 g, 2.83 mmol) in CH2Cl2 (30.0 mL) under ice-cooling in
the same manner as described in Section 4.1.4.. The reaction mix-
ture was stirred for 19 h while gradually raising its temperature to
room temperature, and then water was added thereto under cool-
ing with ice and the mixture was extracted with CH2Cl2. The or-
ganic layer was washed with brine and dried with MgSO4, and
then the solvent was removed under reduced pressure. The ob-
tained residue was purified by silica gel column chromatography
(AcOEt/hexane = 1:5) to obtain the title compound (527 mg,
71%). 1H NMR (CDCl3) d: 3.92 (2H, s), 4.62 (1H, s), 6.73–6.76 (1H,
m), 6.85–6.99 (1H, m), 7.06–7.14 (4H, m), 7.35–7.40 (2H, m).
4.1.14. [2-(4-Isopropylbenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6e)
This compound was prepared from 9 and 2-(4-isopropylben-
zyl)phenol (11e) in the same manner as described in Section
4.1.10. Yield 27%. 1H NMR (CD3OD) d: 0.94 (1H, dd, J = 12.0,
12.9 Hz), 1.22 (6H, d, J = 6.9 Hz), 1.48–1.58 (1H, m), 2.02–2.09
(1H, dt, J = 3.9, 4.2 Hz), 2.78–2.87 (1H, m), 3.12–3.24 (2H, m),
3.43–3.49 (2H, m), 3.68 (1H, dd, J = 4.2, 6.6 Hz), 3.89 (1H, d,
J = 14.4 Hz), 4.01 (1H, d, J = 14.7 Hz), 4.13–4.21 (1H, m), 6.84 (1H,
t, J = 6.3 Hz), 7.02 (1H, d, J = 6.3 Hz), 7.06–7.15 (6H, m); MS (ESI)
m/z: 387 ([M+H]+); HRMS calcd for C23H29O5 ([MÀH]À) 385.2010.
Found 385.2006.
4.1.16.3. [2-(4-Bromobenzyl)phenyl]-5a-carba-b-
D-glucopyrano-
side (14). In nitrogen stream, solution of 13 (386 mg,
a
a
1.46 mmol) in DMF (1 mL) was cooled in an ice bath, and NaH
(60%, 52 mg) was added thereto. The reaction mixture was stirred
at the same temperature for 10 min, and then this reaction mixture
was added dropwise to a suspension of 12 (655 mg, 0.97 mmol) in
DMF (2.5 mL) at À40 °C. The reaction mixture was stirred at the
same temperature for 2 min, at À40 °C for 30 min and then at
0 °C for 1 h. To the reaction mixture was added brine and water.
The resulting solution was extracted with Et2O, and the organic
layer was washed with water and brine and dried over Na2SO4.
The residue obtained by removing the solvent was roughly purified
by silica gel column chromatography (AcOEt/hexane = 1:5) to ob-
tain a crude product (200 mg). The obtained crude product was
dissolved in CH2Cl2 (2.5 mL) in a nitrogen stream. Me2S (0.6 mL,
4.1.15. [2-(4-Cyclopropylbenzyl)phenyl]-5a-carba-b-D-glucopy-
ranoside (6f)
In
a nitrogen stream, 9 (557 mg, 1.03 mmol) and nBu3P
(0.37 mL, 1.55 mmol) were added to a solution of 2-(4-cyclopro-
pylbenzyl)phenol (11f) (348 mg, 1.55 mmol) in toluene (3.5 mL)
under cooling with ice, and then TMAD (267 mg, 1.55 mmol) was
added thereto at the same temperature. The reaction mixture
was stirred for 15 h while gradually raising its temperature to
room temperature. The reaction mixture was concentrated under
reduced pressure, and the obtained residue was treated with silica
gel column chromatography (AcOEt/hexane = 1:10). The obtained
product was dissolved in CH2Cl2 (3.5 mL). Me2S (1.3 mL,
30.3 mmol) and BF3ÁEt2O (0.65 mL, 5.1 mmol) were added thereto
under cooling with ice. The reaction mixture was stirred for 14 h
while gradually raising its temperature to room temperature, and
then water was added thereto under cooling with ice and the mix-
ture was extracted with CH2Cl2. The organic layer was washed
with brine and dried with MgSO4, and then the solvent was re-
moved under reduced pressure. The obtained residue was purified
by silica gel column chromatography (MeOH/CH2Cl2 = 1:9) to
obtain the title compound (98 mg, 25%) as a colorless amorphous
solid. 1H NMR (CD3OD) d: 0.57–0.61 (2H, m), 0.81–0.95 (3H, m),
1.50 (1H, s), 1.79–1.85 (1H, m), 1.99–2.03 (1H, m), 3.15–3.33
(2H, m), 3.42–3.48 (2H, m), 3.66–3.72 (1H, m), 3.81–3.99 (2H, q,
J = 14.7 Hz), 4.41–4.85 (1H, m), 6.80–7.15 (8H, m); MS (ESI) m/z:
407 ([M+Na]+); HRMS calcd for C23H27O5 ([MÀH]À) 383.1853.
Found 383.1849.
13.48 mmol) and BF3ÁEt2O (284
lL, 2.2 mmol) were added thereto
under cooling with ice. The reaction mixture was stirred for 25 h
while gradually raising its temperature to room temperature, then
water was added thereto under cooling with ice and the mixture
was extracted with CH2Cl2. The organic layer was washed with
brine and dried with MgSO4, and then the solvent was removed
under reduced pressure. The obtained residue was purified by sil-
ica gel column chromatography (MeOH/CH2Cl2 = 1:10) to obtain
the title product (40.0 mg, 43%). 1H NMR (CD3OD) d: 0.85 (1H,
dd, J = 11.1, 13.2 Hz), 1.44–1.59 (1H, m), 2.03–2.09 (1H, m), 3.13
(1H, d, J = 8.7 Hz), 3.19 (1H, d, J = 5.1 Hz), 3.37–3.45 (2H, m),
3.63–3.68 (1H, dd, J = 3.9, 10.6 Hz), 3.81 (1H, d, J = 14.4 Hz), 4.10
(1H, d, J = 14.4 Hz), 4.09–4.18 (1H, m), 6.79 (1H, t, J = 7.5 Hz),
6.96 (1H, d, J = 9.0 Hz), 7.03–7.13 (4H, m), 7.27–7.32 (2H, m).
4.1.16.4. [2-(4-Vinylbenzyl)phenyl]-5a-carba-b-D-glucopyrano-
4.1.16. [2-(4-Vinylbenzyl)phenyl]-5a-carba-b-
oside (6g)
4.1.16.1. (1S,2R,3S,4R,5R)-2,3,4-Trisbenzyloxy-(5-benzyloxym-
ethyl)cyclohexyl trifluoromethanesulfonate. (12). Pyridine
D
-glucopyran-
side (6g). In a nitrogen stream, 14 (40 mg, 0.094 mmol) was dis-
solved in toluene (2 mL). Tributylvinyltin (36 mg, 0.11 mmol),
Pd(PPh3)4 (2.7 mg) and 2,6-di-tert-butyl-4-methylphenol were
added thereto, and the mixture was refluxed at 110 °C for 15 h.

5340
Y. Ohtake et al. / Bioorg. Med. Chem. 19 (2011) 5334–5341
The reaction mixture was cooled to room temperature and the sol-
vent was removed under reduced pressure. The obtained residue
was purified by preparative thin-layer chromatography (PTLC)
(MeOH/CH2Cl2 = 1:10) to obtain the title product (9 mg, 26%) as a
colorless amorphous solid. 1H NMR (CD3OD) d: 0.86 (1H, dd,
J = 11.7, 12.0 Hz), 1.42–1.55 (1H, m), 1.96–2.03 (1H, m),
3.10–3.19 (2H, m), 3.36–3.44 (2H, m), 3.63 (1H, dd, J = 4.1,
10.6 Hz), 3.81–4.08 (2H, m), 4.09–4.16 (1H, m), 5.07 (1H, dd,
J = 1.2, 11.0 Hz), 5.63 (1H, dd, J = 1.2, 18.0 Hz), 6.61 (1H, dd,
J = 11.0, 18.0 Hz), 6.78 (1H, t, J = 7.3 Hz), 7.03 (1H, d, J = 6.3 Hz),
7.02–7.24 (6H, m); MS (ESI) m/z: 370 ([M]+); HRMS calcd for
obtain the title compound (66 mg, 74%). 1H NMR (CDCl3) d: 1.60–
1.80 (2H, m), 1.95–2.09 (1H, m), 2.66 (1H, d, J = 4. 8 Hz), 3.38–3.79
(5H, m), 3.65 (1H, s), 3.69 (2H, s), 4.33–4.93 (9H, m), 5.96–6.16
(1H, m), 6.73–7.42 (28H, m); MS (ESI) m/z: 773 ([M+Na]+).
4.1.17.5. [2-(4-Methoxybenzyl)phenyl]-5a-carba-b-D-glucopy-
ranoside (6h). To a solution of 18 (66 mg, 0.0879 mmol) in 0.1 M
HCl–MeOH (2 mL) was added 20% Pd(OH)2 (10 mg), and the reac-
tion mixture was stirred under a hydrogen atmosphere for 3 h.
After the reaction, the reaction mixture was filtered and the filtrate
was concentrated under reduced pressure, and the obtained resi-
due was purified by PTLC (MeOH/CH2Cl2 = 1:10) to obtain the title
compound (20 mg, 61%) as a colorless amorphous solid. 1H NMR
(CD3OD) d: 0.89–1.03 (1H, m), 1.40–1.60 (1H, m), 2.02–2.10 (1H,
m), 3.18–3.34 (2H, m), 3.45–3.51 (2H, m), 3.67–3.71 (1H, m),
3.73 (3H, s), 3.82–3.99 (2H, m), 4.13–4.22 (1H, m), 6.78 (2H, d,
J = 8.6 Hz), 6.80–6.86 (1H, m), 6.99–7.16 (5H, m); MS (ESI) m/z
397 ([M+Na]+); HRMS calcd for C21H25O6 ([MÀH]À) 373.1646.
Found 373.1651.
C
22H25O5 ([MÀH]À) 369.1697. Found 369.1698.
4.1.17. [2-(4-Methoxybenzyl)phenyl]-5a-carba-b-
D
-glucopyran-
oside (6h)
4.1.17.1.
2-(2,3,4,6-Tetra-O-benzyl-5a-carba-b-D-gluco-pyran-
osyl)benzoic acid methyl ester (15). In a nitrogen stream, 9
(200 mg, 0.371 mmol) and Ph3P (146 mg, 0.557 mmol) were added
to a solution of methyl salicylate (72
l
L, 0.557 mmol) in THF
L,
(400 L), and then diethyl azodicarboxylate (DEAD, 88
l
l
0.557 mmol) was added dropwise thereto at room temperature.
The mixture solution was stirred at room temperature for 10 h.
The reaction mixture was concentrated under reduced pressure,
and the obtained residue was purified by PTLC (AcOEt/hex-
ane = 1:3) to obtain the title compound (123 mg, 49%). 1H NMR
(CDCl3) d: 1.60–1.80 (2H, m), 2.15–2.24 (1H, m), 3.48–3.64 (5H,
m), 3.83 (3H, s), 4.43 (3H, s), 4.53 (1H, d, J = 10.7 Hz), 4.51–4.98
(5H, m), 6.95–7.02 (1H, m), 7.10–7.50 (22H, m), 7.78 (1H, dd,
J = 1.7, 7.8 Hz); MS (ESI) m/z: 695 ([M+Na]+).
4.1.18. [2-(4-Ethoxybenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6i)
This compound was prepared from 9 and 2-(4-ethoxyben-
zyl)phenol (11i) in the same manner as described in Section
4.1.10. Yield 36%. 1H NMR (CD3OD) d: 0.89–1.03 (1H, m), 1.35
(3H, t, J = 7.1 Hz), 1.48–1.62 (1H, m), 2.02–2.11 (1H, m), 3.18–
3.33 (3H, m), 3.44–3.52 (2H, m), 3.68–3.74 (1H, m), 3.81–4.02
(3H, m), 4.13–4.23 (1H, m), 6.74–6.87 (4H, m), 6.99–7.17 (4H,
m); MS (ESI) m/z: 411 ([M+Na]+); HRMS calcd for C22H27O6
([MÀH]À) 387.1802. Found 387.1797.
4.1.17.2. 2-(2,3,4,6-Tetra-O-benzyl-5a-carba-b-D-glucopyrano-
syl)benzyl alcohol (16). Lithium aluminum hydride (10.4 mg,
0.274 mmol) was added to a solution of 15 (123 mg, 0.183 mmol)
4.1.19. [2-(4-Methylsulfanylbenzyl)phenyl]-5a-carba-b-D-
glucopyranoside (6j)
in THF (360
l
L) in small portions and the reaction mixture was stir-
This compound was prepared from 9 and 2-(4-methylsulfanylb-
enzyl)phenol (11j) in the same manner as described in Section
4.1.15. Yield 28%. 1H NMR (CD3OD) d: 0.80–0.95 (1H, m), 1.45–
1.60 (1H, m), 2.00–2.10 (1H, m), 2.42 (3H, s), 3.12–3.33 (2H, m),
3.40–3.50 (2H, m), 3.63–3.70 (1H, m), 3.86 (1H, d, J = 15.0 Hz),
4.00 (1H, d, J = 15.0 Hz), 4.15–4.23 (1H, m), 6.84 (1H, t,
J = 7.0 Hz), 7.00–7.18 (7H, m); MS (ESI) m/z: 391 ([M+H]+); HRMS
calcd for C21H25O5S ([MÀH]À) 389.1417. Found 389.1415.
red at 55 °C for 3 h. The reaction mixture was cooled to room tem-
perature, and then water was added thereto and extracted with
AcOEt. The organic layer was washed with brine and dried with
MgSO4, and then the solvent was removed under reduced pressure.
The obtained residue was purified by PTLC (AcOEt/hexane = 1:3) to
obtain the title compound (95 mg, 81%). 1H NMR (CDCl3) d: 1.60–
1.80 (2H, m), 2.16–2.22 (1H, m), 3.46–3.74 (5H, m), 4.43 (3H, s),
4.54 (1H, d, J = 10.7 Hz), 4.66 (2H, br s), 4.81–4.96 (6H, m), 6.94–
7.31 (24H, m); MS (ESI) m/z: 667 ([M+Na]+).
4.1.20. [2-(4-Hydroxybenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6k)
4.1.17.3. 2-(2,3,4,6-Tetra-O-benzyl-5a-carba-b-
D
-glucopyrano-
In a nitrogen stream, 1.0 M BBr3 in CH2Cl2 (0.80 mL, 0.80 mmol)
was added to a solution of 6h (100 mg, 0.27 mmol) in CH2Cl2
(1.3 mL) at À78 °C. The cooling bath was removed and the reaction
mixture was stirred at room temperature for 1 h. The reaction mix-
ture was concentrated under reduced pressure and the obtained
residue was purified by reverse phase column chromatography
(MeOH/20 mM AcONH4 aqueous solution = 5:95–50:50) to obtain
the title compound (45 mg, 47%) as a colorless amorphous solid.
1H NMR (CD3OD) d: 0.92–1.06 (1H, m), 1.47–1.67 (1H, m), 2.03–
2.11 (1H, m), 3.18–3.34 (2H, m), 3.45–3.52 (2H, m), 3.71 (1H, dd,
J = 4.1, 10.7 Hz), 3.79–3.95 (2H, m), 4.12–4.22 (1H, m), 6.65 (2H,
d, J = 8.4 Hz), 6.83 (1H, t, J = 7.3 Hz), 6.99–7.16 (5H, m); MS (ESI)
m/z: 361 ([M+H]+); HRMS calcd for C20H23O6 ([MÀH]À) 359.1489.
Found 359.1481.
syl)benzaldehyde (17). To a solution of 16 (95 mg, 0.147 mmol)
in CH2Cl2 (1.5 mL) was added Dess–Martin reagent (94 mg,
0.221 mmol), and the mixture solution was stirred at room tem-
perature for 45 min. Insolubles were removed from the reaction
mixture by filtration and the filtrate was concentrated under re-
duced pressure. The obtained residue was purified by PTLC
(AcOEt/hexane = 1:3) to obtain the title compound (77 mg, 81%).
1H NMR (CDCl3) d: 1.60–1.80 (2H, m), 2.17–2.23 (1H, m), 3.48–
3.78 (5H, m), 4.44 (3H, s), 4.54 (1H, d, J = 10.7 Hz), 4.73–4.97
(5H, m), 7.00–7.32 (22H, m), 7.50 (1H, dd, J = 1.5, 7.8 Hz), 7.83
(1H, dd, J = 1.5, 7.6 Hz), 10.4 (1H, s).
4.1.17.4. [2-(2,3,4,6-Tetra-O-benzyl-5a-carba-b-
osyl)phenyl]-4-methoxyphenylmethanol (18). To a solution of
17 (77 mg, 0.119 mmol) in Et2O (120 L) was added 0.5 M
4-methoxyphenyl-magnesiumbromide in THF solution (480 L,
D-gluco-pyran-
l
4.1.21. [2-(4-Fluorobenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6l)
l
0.238 mmol), and the mixture solution was stirred at room temper-
ature for 13 h. A saturated aq NH4Cl was added thereto and the mix-
ture was extracted with AcOEt. The organic layer was dried with
MgSO4 and the solvent was concentrated under reduced pressure.
The obtained residue was purified by PTLC (AcOEt/hexane = 1:3) to
This compound was prepared from 9 and 2-(4-fluoroben-
zyl)phenol (11l) in the same manner as described in Section
4.1.10. Yield 46%. 1H NMR (CD3OD) d: 0.99 (1H, m), 1.55 (1H, m),
2.08 (1H, m), 3.22 (1H, m), 3.30 (1H, m), 3.48 (1H, m), 3.51 (1H,
dd, J = 6.0, 10.8 Hz), 3.70 (1H, dd, J = 4.2, 10.8 Hz), 3.90 (1H, d,

Y. Ohtake et al. / Bioorg. Med. Chem. 19 (2011) 5334–5341
5341
J = 14.7 Hz), 4.02 (1H, d, J = 14.7 Hz), 4.20 (1H, ddd, J = 4.8, 9.0,
11.4 Hz), 6.85 (1H, t, J = 7.5 Hz), 6.93 (1H, m), 7.02 (1H, d,
J = 8.1 Hz), 7.06–7.23 (5H, m); MS (ESI) m/z: 363 ([M+H]+); HRMS
calcd for C20H22FO5 ([MÀH]À) 361.1446. Found 361.1441.
6h), and 24 h after administration. The drug concentration in plas-
ma sample was measured using an LC–MS/MS system after depro-
teinization. LC–MS/MS analysis was performed using a Shimadzu
LC-10AD pump and an Applied Biosystems/MDS Sciex API-300
mass spectrometer with a TurboIonSpray source. Chromatographic
separation was achieved using a Shiseido CAPCELL PAK C18 col-
4.1.22. [2-(4-Chlorobenzyl)phenyl]-5a-carba-b-D-glucopyran-
oside (6m)
umn (2.0 Â 150 mm, 5
lm). The mobile phase consisted of acetoni-
This compound was prepared from 9 and 2-(4-chloroben-
zyl)phenol (11m) in the same manner as described in Section
4.1.15. Yield 35%. 1H NMR (CD3OD) d: 0.95 (1H, dd, J = 11.1,
13.2 Hz), 1.53 (1H, m), 2.00–2.09 (1H, m), 3.20 (1H, d, J = 8.7 Hz),
3.25 (1H, d, J = 5.1 Hz), 3.43–3.51 (2H, m), 3.68–3.71 (1H, m),
3.91 (1H, d, J = 14.7 Hz), 3.99 (1H, d, J = 15.0 Hz), 4.15–4.23 (1H,
m), 6.85 (1H, t, J = 7.5 Hz), 7.03 (1H, d, J = 9.0 Hz), 7.08–7.23 (6H,
m); MS (ESI) m/z: 379 ([M+H]+); HRMS calcd for C20H22ClO5
([MÀH]À) 377.1150. Found 377.1152.
trile/10 mM ammonium acetate solution (4:6, v/v) and the flow
rate was set at 0.2 mL/minute. The injection cycle of each sample
was set at 10 min. The transitions for multiple reaction monitoring
were 394–180 (3-active), 392–267 (6h). Non-compartmental
pharmacokinetic parameters were calculated based on the
averaged plasma concentration-time data using WinNonlin
Professional 5.0 (Pharsight, Mountain View, CA).
Acknowledgements
4.1.23. [2-(4-Trifluoromethylbenzyl)phenyl]-5a-carba-b-
pyranoside (6n)
D
-gluco
We are very thankful to Ms. Hitomi Suda of Chugai Pharmaceu-
tical Co., Ltd for HRMS measurements of the compounds, and Edit-
ing Services at Chugai Pharmaceutical Co., Ltd for assistance with
English usage.
This compound was prepared from 9 and 2-(4-trifluorometh-
ylbenzyl)phenol (11n) in the same manner as described in Section
4.1.10. Yield 19%. 1H NMR (CD3OD) d: 0.89 (1H, dd, J = 11.7,
12.9 Hz), 1.48–1.61 (1H, m), 2.05 (1H, dt, J = 13.2, 4.2 Hz),
3.15–3.34 (2H, m), 3.42–3.49 (2H, m), 3.69 (1H, dd, J = 3.9,
10.5 Hz), 3.96–4.15 (2H, m), 4.17–4.24 (1H, m), 6.87 (1H, dt,
J = 1.2, 7.5 Hz), 7.02–7.20 (3H, m), 7.38 (2H, d, J = 8.1 Hz), 7.51
(2H, d, J = 8.4 Hz); MS (ESI) m/z: 435 ([M+Na]+); HRMS calcd for
References and notes
1. You, G.; Lee, W. S.; Barros, E. J. G.; Kanai, Y.; Huo, T. L.; Khawaja, S.; Wells, R. G.;
Nigam, S. K.; Hediger, M. A. J. Biol. Chem. 1995, 270, 29365.
2. Wright, E. M.; Turk, E.; Martin, M. G. Cell Biochem. Biophys. 2002, 36, 115.
3. Nomura, S. Curr. Top. Med. Chem. 2010, 10, 411.
4. Ehrenkanz, J. R. L.; Lewis, N. G.; Kahn, C. R.; Roth, J. Diabates/Metab. Res. Rev.
2005, 21, 31.
C
21H22F3O5 ([MÀH]À) 411.1414. Found 411.1410.
5. (a) Oku, A.; Ueta, K.; Arakawa, K.; Ishihara, T.; Nawano, M.; Kuronuma, Y.;
Matsumoto, M.; Saito, A.; Tsujihara, K.; Anai, M.; Asano, T.; Kanai, Y.; Endou, H.
Diabetes 1999, 48, 1794; (b) Tsujihara, K.; Hongu, M.; Saito, K.; Kawanishi, H.;
Kuriyama, K.; Matsumoto, M.; Oku, A.; Ueta, K.; Tsuda, M.; Saito, A. J. Med.
Chem. 1999, 42, 5311.
5. Biology
5.1. In vitro SGLT inhibition assay
6. (a) Fushimi, N.; Ito, F.; Isaji, M. PCT Int. Appl. WO2003011880, 2004.; (b)
Katsuno, K.; Fujimori, Y.; Takemura, Y.; Hiratochi, M.; Itoh, F.; Komatsu, Y.;
Fujikura, H.; Isaji, M. J. Pharmacol. Exp. Ther. 2007, 320, 323.
7. Fujimori, Y.; Katsuno, K.; Nakashima, I.; Ishikawa-Takemura, Y.; Fujikura, H.;
Isaji, M. J. Pharmacol. Exp. Ther. 2008, 327, 268.
Chinese hamster ovary-K1(CHO) cells stably expressing human
SGLT2 (NM_003041) and human SGLT1 (NM_000343) were used
for the sodium-dependent methyl-a-D-glucopyranoside (AMG) up-
take inhibition assay. The cells were incubated in reaction buffer with
compound and1 mMAMG containing [14C]AMGfor45 min. AMG up-
take activities were determined by counting the radioactivity of the
cell lysates. IC50 values were calculated by curve fitting using a
four-parameter logistic model (XLfit, ID Business Solutions Ltd.).
8. (a) 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, W. N.; Janovitz, E. B.; Flint, O. P.; Whaley, J. M.; Washburn, W. N. J.
Med. Chem. 2008, 51, 1145; (b) Washburn, W. N. J. Med. Chem. 2009, 52, 1785.
9. Nomura, S.; Sakamaki, S.; Hongu, M.; Kawanishi, E.; Koga, Y.; Sakamoto, T.;
Yamamoto, Y.; Ueta, K.; Kimata, H.; Nakayama, K.; Tsuda-Tsukimoto, M. J. Med.
Chem. 2010, 53, 6355.
5.2. In vivo blood glucose lowering test in db/db mice
10. (a) Sato, M.; Kakinuma, H.; Asanuma, H. PCT Int. Appl. WO2004014931, 2004;
Chem. Abstr. 2004, 140, 199631.; (b) Kakinuma, H.; Oi, T.; Hashimoto-Tsuchiya,
Y.; Arai, M.; Kawakita, Y.; Fukasawa, Y.; Iida, I.; Hagima, N.; Takeuchi, H.; Chino,
Y.; Asami, J.; Okumura-Kitajima, L.; Io, F.; Yamamoto, D.; Miyata, N.; Takahashi,
T.; Uchida, S.; Yamamoto, K. J. Med. Chem. 2010, 53, 3247; (c) Goodwin, N. C.;
Mabon, R.; Harrison, B. A.; Shadoan, M. K.; Almstead, Z. Y.; Xie, Y.; Healy, J.;
Buhring, L. M.; DaCosta, C. M.; Bardenhagen, J.; Mseeh, F.; Liu, Q.; Nouraldeen,
A.; Wilson, A. G. E.; Kimball, S. D.; Powell, D. R.; Rawlins, D. B. J. Med. Chem.
2009, 52, 6201; (d) Robinson, R. P.; Mascitti, V.; Boustany-Kari, C. M.; Carr, C. L.;
Foley, P. M.; Kimoto, E.; Leininger, M. T.; Lowe, A.; Klenotic, M. K.; MacDonald, J.
I.; Maguire, R. J.; Masterson, V. M.; Maurer, T. S.; Miao, Z.; Patel, J. D.; Préville,
C.; Reese, M. R.; She, L.; Steppan, C. M.; Thuma, B. A.; Zhu, T. Bioorg. Med. Chem.
Lett. 2010, 20, 1569.
Male db/db mice were purchased from CLEA Japan (Tokyo,
Japan) and maintained on a regular diet (CE2, CLEA Japan). The
animals received a single oral dose of 3 and 6h (100 mg/kg) formu-
lated as homogenous suspensions in 0.5% carboxymethyl-cellulose
sodium salt via oral gavage. Blood samples were collected just
before and at 1, 2, 4, 6, 8, 10, and 24 h after dosing. Blood glucose
levels were measured by the hexokinase method (Autosera S GLU,
Daiichi Pure Chemicals, Japan).
11. Arjona, O.; Gómez, A. M.; López, J. C.; Plumet, J. Chem. Rev. 2007, 107, 1919.
12. Matsuoka, H.; Sato, T.; Nishimoto, M.; Shimma N. PCT Int. Appl.
WO2006011469, 2006; EP1783110.
5.3. Pharmacokinetics Studies
The db/db mice were administered a drug, and subjected to
blood sampling under the same conditions as the animals used
for the in vivo blood glucose lowering test. Animals received a sin-
gle oral administration of 3 and 6h at a dose of 10 mg/kg (n = 6).
Blood samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8, 10 (for
13. Tsunoda, H.; Ogawa, S. Liebigs Ann. 1995, 2, 267.
14. Sudha, A. V. R. L.; Nagarajan, M. Chem. Commun. 1998, 925.
15. Huston, R. C.; Neeley, A.; Fayerweather, B. L.; D’Arcy, H. M.; Maxfield, F. H.;
Ballard, M. M.; Lewis, W. C. J. Am. Chem. Soc. 1933, 55, 2146.
16. Frick, W.; Glombik, H.; Kramer, W.; Heuer, H.; Brummerhop, H.; Plettemburg,
O. PCT Int. Appl. WO04052902, 2004.