Discovery of canagliflozin, a novel C-glucoside with thiophene ring, as sodium-dependent glucose cotransporter 2 inhibitor for the treatment of type 2 diabetes mellitus (1)

Article abstract of DOI:10.1021/jm100332n

We discovered that C-glucosides 4 bearing a heteroaromatic ring formed metabolically more stable inhibitors for sodium-dependent glucose cotransporter 2 (SGLT2) than the O-glucoside, 2 (T-1095). A novel thiophene derivative 4b-3 (canagliflozin) was a highly potent and selective SGLT2 inhibitor and showed pronounced anti-hyperglycemic effects in high-fat diet fed KK (HF-KK) mice.

Full text of DOI:10.1021/jm100332n

J. Med. Chem. 2010, 53, 6355–6360 6355
DOI: 10.1021/jm100332n
Discovery of Canagliflozin, a Novel C-Glucoside with Thiophene Ring, as Sodium-Dependent
Glucose Cotransporter 2 Inhibitor for the Treatment of Type 2 Diabetes Mellitus1
Sumihiro Nomura,*,† Shigeki Sakamaki,† Mitsuya Hongu,† Eiji Kawanishi,† Yuichi Koga,† Toshiaki Sakamoto,†
Yasuo Yamamoto,† Kiichiro Ueta,‡ Hirotaka Kimata,‡ Keiko Nakayama,‡ and Minoru Tsuda-Tsukimoto§
†Medicinal Chemistry Research Laboratories, ‡Pharmacology Research Laboratories, and §DMPK Research Laboratories,
Mitsubishi Tanabe Pharma Corporation, 2-2-50 Kawagishi, Toda, Saitama, Japan
Received March 11, 2010
We discovered that C-glucosides 4 bearing a heteroaromatic ring formed metabolically more stable
inhibitors for sodium-dependent glucose cotransporter 2 (SGLT2) than the O-glucoside, 2 (T-1095).
A novel thiophene derivative 4b-3 (canagliflozin) was a highly potent and selective SGLT2 inhibitor and
showed pronounced anti-hyperglycemic effects in high-fat diet fed KK (HF-KK) mice.
Introduction
Orally active 2 is an ester prodrug of active metabolite 1 (T-
1095A), which enhances the resistance against hydrolysis by
β-glucosidase in the intestine.10 Nevertheless, the O-glucoside
part of 2 is at least hydrolyzed to its aglycon in vivo (data not
shown). The C-glucoside 3 was disclosed as SGLT2 inhibitor
by Bristol-Myers Squibb Co.17,18 Accordingly, to explore
novel C-glucosides metabolically more stable than O-gluco-
sides, we evaluated a series of C-glucosides 4 bearing a
heteroaromatic ring.
The incidence of type 2 diabetes mellitus (T2DMa) is
markedly increasing in Westernized societies and some devel-
oping countries.2-4 However, at present, no single agent is
capable of achieving acceptable, long-lasting blood glucose
control in the majority of patients.5 Accordingly, there is a
strong incentive to develop novel drugs with improved effi-
cacy and safety.
Plasma glucose is filtered in the glomerulus and then
reabsorbed in the proximal tubules in the kidney. Renal
glucose reabsorption is mediated predominantly by SGLT2
and to a lesser extent by SGLT1.6-8 In the normoglycemic
state, all filtered glucose is transported from the tubular lumen
to the blood. However, under hyperglycemic conditions, the
reabsorption process is saturated and urinary glucose excre-
tion (UGE) increases linearly.9 An SGLT2 inhibitor, 2 (T-
1095, Figure 1), enhanced UGE and consequently lowered
blood glucose levels in diabetic animal models independent of
insulin action.10,11 SGLT1 is distributed in the intestine, heart
and trachea besides the kidney, while SGLT2 is located solely
in the kidney.12 Therefore, selective SGLT2 inhibitors would
be desirable for anti-diabetic agents. Given that SGLT2 inhi-
bition lowers plasma glucose levels in an insulin-independent
fashion, SGLT2 inhibitors are not predicted to be associated
with hypoglycemia, which is a major concern for the current
therapieswithinsulinandsulfonylureas. Inaddition, increases
in urinary caloric loss due to UGE predict that SGLT2
inhibitors will not be associated with weight gain that is often
seen with other classes of currently approved antihyperglyce-
mic agents. As a consequence of these anticipated properties,
identification of novel SGLT2 inhibitors became a goal for
medicinal chemistry.13-16
Chemistry
We describe herein the syntheses of C-glucosides 4a-4e
bearing a heteroaromatic ring. The synthetic route is outlined
in Scheme 1. We took advantage of the synthetic strategy
outlined by Deshpande et al.19-21 Aglycons 5a-5d were dis-
solved in tetrahydrofuran and toluene, and treated with
n-butyllithium at -78 °C to generate aryllithium, followed by
addition of 2,3,4,6-tetra-O-trimethylsilyl-β-D-gluconolactone.22
Aglycon 5e underwent deprotonation with 1 equiv of n-butyl-
lithium at the benzylic position rather than lithium-bromine
exchange. Additional amounts of n-butyllithium did not induce
lithiation at all. The aryllithium of 5e was generated by
tert-butyllithium as the second equivalent. The resulting anome-
ric mixture of lactols was immediately converted into desilylated
methyl ethers 6a-6e by addition of methanesulfonic acid in
methanol. Finally, C-glucoside derivatives 4a-4e were obtained
by stereoselective reduction of 6a-6e using a combination of
triethylsilane and boron trifluoride etherate in methylene
chloride.23,24 The stereochemistry of 4a-4e was determined as
β-configuration by the coupling constant between anomeric
C-H and adjacent C-H (J ≈ 9.5 Hz) in the 1H NMR
spectrum.
Synthetic routes to aglycons 5a-5e are shown in Scheme 2.
Aglycon 5a-1 having furan was prepared by a coupling
reaction of bromophenyllithium and furaldehyde in diethyl
ether followed by reduction of hydroxyl group with iodotri-
methylsilane, in situ generated by chlorotrimethylsilane and
sodium iodide in acetonitrile.25 Thiophene aglycons 5b-1, 2and 3
were synthesized by Friedel-Crafts acylation of corresponding
*To whom correspondence should be addresses. Phone: þ81-48-433-2503.
Fax: þ81-48-433-8150. E-mail: nomura.sumihiro@mw.mt-pharma.co.jp.
aAbbreviations: SGLT, sodium-dependent glucose cotransporter;
HF-KK, high-fat diet fed KK; T2DM, type 2 diabetes mellitus; UGE,
urinary glucose excretion; SD, Sprague-Dawley; SAR, structure-
activity relationship; GLUT, facilitated glucose transporter; AMG,
R-methyl-D-glucopyranoside; CHOK, Chinese hamster ovary-K;
2-DG, 2-deoxy-D-glucose.
r
2010 American Chemical Society
Published on Web 08/06/2010
pubs.acs.org/jmc

6356 Journal of Medicinal Chemistry, 2010, Vol. 53, No. 17
Nomura et al.
Figure 1. Structures of some O- and C-glucosides.
Scheme 1. Syntheses of 4a-4ea
a Reagents and conditions: (a) n-BuLi, THF-toluene, -78 °C; (b) n-BuLi, then tert-BuLi, THF-toluene, -78 °C; (c) 2,3,4,6-tetra-O-trimethylsilyl-
β-D-gluconolactone, toluene, -78 °C; (d) MeSO3H, MeOH; (e) Et3SiH, BF3 OEt2, CH2Cl2, -78 to 0 °C.
3
Scheme 2. Syntheses of Aglycons 5a-5ea
a Reagents and conditions: (a) n-BuLi, Et2O, -78 °C, then 5-ethyl-2-furaldehyde, -78 °C; (b) TMSCl, NaI, CH3CN, 0 °C; (c) AlCl3, CH2Cl2, 0 °C;
(d) Et3SiH, BF3 OEt2, CH2Cl2, 0 °C; (e) n-BuLi, Et2O, -78 °C, then 5-bromo-2-chlorobenzaldehyde, -78 °C; (f ) NaBH(OAc)3, TFA, 0 °C; (g) n-BuLi,
3
Et2O, -78 °C, then 5-bromo-2-chloro-N-methoxy-N-methylbenzamide, -78 °C; (h) Et3Al, Pd(PPh3)4, CeCl3, THF, 30 °C; (i) NH2NH2 H2O, KOH,
3
ethylene glycol, 190 °C; ( j) 2-amino-40-fluoroacetophenone hydrochloride, EDC, HOBt, Et3N, CH2Cl2; (k) Lawesson’s reagent, 1,4-dioxane, 100 °C.
benzoyl chloride and thiophene in methylene chloride, followed
by reduction of ketone with triethylsilane and boron tri-
fluoride etherate in methylene chloride. Bromine-substituted
phenylpyrazole26 was lithiated with n-butyllithium and reacted
with bromobenzaldehyde in diethyl ether, and the resulting
alcohol was reduced using sodium triacetoxyborohydride in
trifluoroacetic acid27 to give pyrazole derivative 5c-1. Pyridinyl
aglycon 5d-1 was synthesized by a coupling reaction of Weinreb
amide and pyridyllithium28 in diethyl ether, followed by a
cross-coupling of triethylaluminum with bromopyridine in

Article
Journal of Medicinal Chemistry, 2010, Vol. 53, No. 17 6357
Table 1. SAR of the Representative C-Glucosides with Heteroaromatic
Ringa
Table 3. Pharmacokinetic (PK) Parameters of 1 and 4b-3 in Male
Sprague-Dawley (SD) Rats Following Intravenous and Oral Admin-
istrations
compd
1
2a
4b-3
4b-3
10
dose (mg/kg)
route
1
10
3
iv
po
80
iv
po
Cmax (ng/mL)
tmax (h)
AUC0-inf (ng h/mL)
2513
5.0
1.3
304
2.2
153
1.3
12703
5.0
236
35980
5.2
3
t
1/2 (h)
CLtot (mL/h/kg)
Vdss (mL/kg)
F (%)
7506
11390
1357
20
85
a Because a prodrug 2 is very rapidly converted to 1 in vivo, PK
parameters of 1 are shown.
optimization. For R1 substituents, halogeno (chloro of 4b-2) or
lower alkyl (methyl of 4b-3) group provided better in vitro
potency than proton (4b-1). Substituted aryl (4-fluorophenyl of
4b-3) group was preferred to alkyl (ethyl of 4b-1) or halogeno
(chloro of 4b-2) group as R2 substituents for in vivo potency and
chemical stability (data not shown). From these thiophene deri-
vatives, compound 4b-3 (canagliflozin, TA-7284/JNJ-28431754)
was selected as a clinical candidate.
Table 2 displays effects of 1, 2, and 4b-3 on hSGLT1,
hSGLT2, facilitated glucose transporter 1 (GLUT1) activity
and UGE in rats. The inhibitory effects of 1 and 4b-3 on the
uptake of [14C]R-methyl-D-glucopyranoside (AMG) were in-
vestigated in Chinese hamster ovary-K (CHOK) cells stably
expressing either hSGLT1 or hSGLT2. IC50 values of 1 and 4b-
3 were 240 and 910 nM for hSGLT1, and 5.2 and 2.2 nM for
hSGLT2, respectively. Since GLUT1 mediates the uptake of
glucose in almost all tissues, selectivity of versus GLUT1 was
determined using L6 myoblast cells. Neither compound inhi-
bited the incorporation of [3H]2-deoxy-D-glucose (2-DG)
mediated by GLUT1 predominantly expressed in L6 myo-
blast cells.30 Namely, compound 4b-3 was identified as a
potent and selective inhibitor of hSGLT2.
a Each compound was orally administered at a dose of 30 mg/kg
to male Sprague-Dawley (SD) rats. Urinary glucose excretion (UGE)
data over 24 h were normalized per 200 g body weight. b N.D.: not
determined.
Table 2. hSGLT1, hSGLT2, Facilitated Glucose Transporter
(GLUT1) Inhibitory Activity, and Rat Urinary Glucose Excretion
1
(UGE) Data for 1, 2, and 4b-3
IC50 (nM)
compd
hSGLT1
240
hSGLT2
5.2
GLUT1
>10000
UGEa (mg/day)
Oral administration at 30 mg/kg of 2 and 4b-3 to male SD
rats induced glucose excretion over 24 h by 422 and 3,696 mg,
respectively, per 200 g body weight. Pharmacokinetic studies
revealed a much higher exposure of 4b-3 following oral admin-
istration (Table 3). Following intravenous and oral doses of 3
1
2b
4b-3
422
3696
910
2.2
>10000
a Compound 2 or 4b-3 was orally administered at a dose of 30 mg/kg
to male SD rats. UGE data over 24 h were normalized per 200 g body
weight. b An ester prodrug 2 is rapidly converted to active metabolite 1 in
vivo, thus in vitro hSGLT inhibitory activities of 2 are not shown.
and 10 mg/kg, respectively, to male SD rats, AUC0-inf,po, t1/2,po,
and oral bioavailability were determined to be 35,980 ng h/mL,
3
5.2 h, and 85%, respectively. Thus, inhibition of SGLT2 in
renal tubules after oral dosing of 4b-3 is likely to continuously
suppress reabsorption of glucose. The extensive UGE would
reflect excellent pharmacokinetic properties of 4b-3 in vivo as
well as high potency of SGLT2 inhibition. Since most of the
filtered glucose is reabsorbed by SGLT2 in the renal tubules,
the novel compound would be useful for an anti-diabetic agent.
Single oral administration of 4b-3 at 3 mg/kg remarkably
reduced blood glucose levels without influencing food intake
in hyperglycemic high-fat diet fed KK (HF-KK) mice
(Figure 2A). There was a 48% reduction in blood glucose
level versus vehicle at 6 h. In contrast, compound 4b-3 only
slightly affected blood glucose levels in normoglycemic mice
(Figure 2B). Therefore, this compound would control hyper-
glycemia in the therapy of T2DM with low risk of hypogly-
cemia.
tetrahydrofuran and reduction of the carbonyl group with
hydrazine hydrate and potassium hydroxide in ethylene
glycol.29 To provide thiazole derivative 5e-1, ketoamide was
cyclized using Lawesson’s reagent in 1,4-dioxane.
Results and Discussion
Effects of furan, thiophene, pyrazole, pyridine and thiazole
derivatives 4 (Figure 1) were evaluated on human SGLT2
(hSGLT2) activity and on urinary glucose excretion (UGE) in
male Sprague-Dawley (SD) rats per 200 g of body weight
over 24 h. The structure-activity relationship (SAR) of the
representative compounds are shown in Table 1. Ethylthio-
phene derivative 4b-1 possessed good hSGLT2 inhibitory po-
tency compared with the corresponding furan 4a-1 or pyridine
4d-1 derivative. Also, phenylthiophene derivative 4b-3 showed
higher hSGLT2 inhibitory activity and UGE effect than the
corresponding pyrazole 4c-1 or thiazole 4e-1 derivative. There-
fore, we selected the novel thiophene derivatives for further
In brief, compound 4b-3 is a highly potent and selective
inhibitor for hSGLT2 with favorable pharmacokinetic profiles,
and remarkably increases urinary glucose excretion. In addi-
tion, oral administration of 4b-3 induced anti-hyperglycemic

6358 Journal of Medicinal Chemistry, 2010, Vol. 53, No. 17
Nomura et al.
and solvents were purchased from commercial suppliers and used
without further purification unless otherwise noted. Reaction
products were monitored by TLC using 0.25 mm E. Merck silica
gel plates (60 F254) and were visualized using UV light or 5%
phosphomolybdic acid in 95% EtOH. NMR spectra were col-
lected on JEOL JNM-ECX400P and Varian UNITY INOVA500
spectrometers. Chemical shiftsaregiven inparts per million (ppm)
downfield from internal reference tetramethylsilane standard;
coupling constants (J value) are given in hertz (Hz). Elemental
analyses were conducted by Medicinal Chemistry Research
Laboratories, Mitsubishi Tanabe Pharma. Melting points were
€
measured by a Buchi model B-545 instrument and were uncor-
rected. Infrared spectra were measured on Perkin-Elmer PARA-
GON1000. APCI-MS spectra were obtained on Finnigan MAT
SSQ7000CorThermoQuestLCQAdvantage, eluting with10mM
AcONH4/MeOH. GC-MS spectra were measured on Shimadzu
GCMS-QP2010. Analytical HPLC spectra were reported using
Agilent 1100 with a UV detector measuring absorbance at 210 nm.
All compounds were found to be >95% pure by HPLC analysis
unless otherwise noted.
1-(β-D-Glucopyranosyl)-4-methyl-3-(5-(4-fluorophenyl)-2-thienyl-
methyl)benzene (4b-3). 5-Bromo-2-methylbenzoic acid17,18 (44.43 g,
206.6 mmol) was suspended in dichloromethane (600 mL), and to
the mixture were added oxalyl chloride (20.0 mL, 229 mmol) and N,
N-dimethylformamide (0.74 mL, 9.55 mmol). The mixture was
stirred at room temperature for 6 h (clear solution). The solvent was
evaporated under reduced pressure to give 5-bromo-2-methyl-
benzoyl chloride as an oil. This compound and 2-(4-fluoro-
phenyl)thiophene31 (36.83 g, 206.6 mmol) were dissolved in
dichloromethane (1,200 mL), and to the mixture was added
aluminum chloride (30.3 g, 227.2 mmol) at 0 °C (internal
temperature). After being stirred at the same temperature for
30 min, the mixture was allowed to warm to room temperature
and stirred overnight. The reaction mixture was poured into
ice-water (1,200 mL). The organic layer was separated, and the
aqueous layer was extracted with chloroform (500 mL) three
times. The organic layers were combined and dried over potas-
sium carbonate. The solvent was concentrated under reduced
pressure, and to the mixture was added n-hexane to produce a
precipitate. The precipitate was collected by filtration, washed
with n-hexane, and dried to give (5-bromo-2-methylphenyl)(5-(4-
fluorophenyl)-2-thienyl)methanone (66.44 g, 85.7%) as yellow crys-
tals: mp 121-122 °C. IR (Nujol) 1668, 1627, 1597, 1585, 1556,
1
1532, 1505 cm-1. APCI-MS m/z 375/377 (M þ H). H NMR
(DMSO-d6) δ 2.24 (3H, s), 7.31-7.37 (3H, m), 7.46 (1H, d, J = 4.0
Hz, thio), 7.63-7.68 (3H, m), 7.87 (2H, m). Anal. Calcd for
C18H12BrFOS: C, 57.61; H, 3.22; Br, 21.29; F, 5.06; S, 8.54. Found:
C, 57.61; H, 3.14; Br, 21.07; F, 4.96; S, 8.58.
A solution of the above obtained (5-bromo-2-methylphenyl)-
(5-(4-fluorophenyl)-2-thienyl)methanone (62.23 g, 165.8 mmol)
and triethylsilane (76.8 mL, 481 mmol) in dichloromethane
(620 mL)-acetonitrile (620 mL) was cooled to 0 °C (internal
temperature), and to the mixture was added dropwise boron
trifluoride-ethyl ether complex (58.8 mL, 464 mmol) over
15 min under nitrogen atmosphere. Subsequently, the mixture
was stirred at room temperature for 4 h and was cooled again under
ice-water. To the mixture was added slowly a saturated aqueous
sodium hydrogen carbonate solution (1,200 mL) under stirring. The
organic layer was separated, and the aqueous layer was extracted
with chloroform (500 mL) three times. The combined organic layer
was washed with brine and dried over magnesium sulfate. The
solvent was evaporated under reduced pressure. The residue was
dissolved in ethyl acetate (1,200 mL)-methanol (600 mL), and the
mixture was treated with activated carbon (3.0 g). The insoluble was
filtered off, and the filtrate was concentrated under reduced pres-
sure. To the mixture was added methanol to produce a precipitate.
The precipitate was collected by filtration, washed with ethyl acetate-
methanol (1:3), and dried to give 2-(5-bromo-2-methylbenzyl)-5-(4-
fluorophenyl)thiophene 5b-3 (46.83 g, 78.2%) as pale-yellow
crystals: mp 101-103 °C. IR (Nujol) 1879, 1746, 1592, 1511
Figure 2. Effects of single oral dosing of 4b-3 on blood glucose
levels and food intake in high-fat diet fed KK (HF-KK) (A) and
normal (B) mice. Data are expressed as the mean ( SEM (n = 5):
* P < 0.05, ** P < 0.01 vs vehicle.
effects in HF-KK mice. Currently, the thiophene derivative
4b-3 (canagliflozin) is being developed for the treatment of
type 2 diabetes mellitus.
Experimental Section
All reactions were carried out under inert gas or with CaCl2
tube, and reaction mixtures werestirred magnetically. Allreagents

Article
Journal of Medicinal Chemistry, 2010, Vol. 53, No. 17 6359
cm-1. GC-MS m/z 362 (Mþ). 1H NMR (DMSO-d6) δ 2.25 (3H, s),
4.15 (2H, s, Ph-CH2-thio), 6.85 (1H, d, J = 3.5 Hz, thio), 7.17 (1H,
d, J = 8.0 Hz), 7.21 (2H, quasi-t), 7.31 (1H, d, J = 3.5 Hz, thio),
7.36 (1H, dd, J = 8.0, 1.9 Hz), 7.44 (1H, d, J = 1.9 Hz), 7.60 (2H,
m). Anal. Calcd for C18H14BrFS: C, 59.84; H, 3.91; Br, 22.12; F,
5.26; S, 8.87. Found: C, 59.89; H, 3.86; Br, 21.93; F, 5.17; S, 8.85.
To a solution of the above obtained 2-(5-bromo-2-methyl-
benzyl)-5-(4-fluorophenyl)thiophene 5b-3 (28.9 g, 80.0 mmol)
in tetrahydrofuran (480 mL) and toluene (480 mL) was added
n-butyllithium (1.6 M n-hexane solution, 50.0 mL, 80.0 mmol)
dropwise over 10 min at -67 to -70 °C (internal temperature)
under argon atmosphere, and the mixture was stirred for 20 min
at the same temperature (dark-blue solution). To the mixture
was added a solution of 2,3,4,6-tetra-O-trimethylsilyl-β-D-glu-
conolactone22 (34.0 g, 72.8 mmol) in toluene (240 mL) dropwise
over 30 min at -67 to -70 °C (internal temperature), and the
mixture was further stirred for 1 h at the same temperature
(slightly brown solution). Subsequently, to the mixture was
added a solution of methanesulfonic acid (21.0 g, 219 mmol)
in methanol (480 mL) dropwise over 15 min, and the resulting
mixture was allowed to warm to room temperature and stirred
for 17 h. The mixture was again cooled in ice-water, and to it
was added a saturated aqueous sodium hydrogen carbonate
solution (1,000 mL). The mixture was extracted with ethyl
acetate (1,000 mL) twice, and the combined organic layer was
washed with brine (1,000 mL) and dried over magnesium sul-
fate. The insoluble was filtered off, and the solvent was evapo-
rated under reduced pressure. The residue was triturated with
toluene (100 mL)-n-hexane (400 mL) to give 1-(1-meth-
oxyglucopyranosyl)-4-methyl-3-(5-(4-fluorophenyl)-2-thienyl-
methyl)benzene 6b-3 (31.6 g, 91.3%) as a pale-yellow powder:
HPLC 88.5% (tR = 8.1 min, L-column ODS (5 μm particle size,
4.6 ꢀ 150 mm), CH3CN/20 mM phosphate buffer (pH 6.5) (45/
55)). APCI-MS m/z 492 (M þ NH4), 460 (M þ NH4 - MeOH),
J = 5.6 Hz, OH), 4.92 (2H, d, J = 4.8 Hz, OH), 6.80 (1H, d, J =
3.5 Hz, thio), 7.11-7.15 (2H, m, Ph), 7.18-7.25 (3H, m, Ph), 7.28
(1H, d, J = 3.5 Hz, thio), 7.59 (2H, dd, J = 8.8, 5.4 Hz, Ph). Anal.
Calcd for C24H25FO5S 0.5H2O: C, 63.56; H, 5.78; F, 4.19; S, 7.07.
3
Found: C, 63.52; H, 5.72; F, 4.08; S, 7.00.
Supporting Information Available: Description of in vitro
hSGLT1, hSGLT2 and GLUT1 assays, and in vivo urinary
glucose excretion and blood glucose-lowering studies; detailed
synthetic procedures for 4a-1, 4b-1, 4b-2, 4c-1, 4d-1 and 4e-1;
HPLC analysis of 4b-3. This material is available free of charge
via the Internet at http://pubs.acs.org.
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1
443 (M þ H - MeOH). H NMR (DMSO-d6) δ 2.26 (3H, s,
Me), 2.91 (1H, m, sugar), 2.95 (3H, s, OMe), 3.21 (1H, m,
sugar), 3.37 (1H, m, sugar), 3.51-3.61 (2H, m, sugar), 3.75 (1H,
m, sugar), 4.09, 4.18 (each 1H, d, J = 15.9 Hz, Ph-CH2-thio),
4.51 (1H, t, J = 6.0 Hz, OH), 4.65 (1H, d, J = 7.2 Hz, OH), 4.69
(1H, d, J= 5.1 Hz, OH), 4.94 (1H, d, J= 5.5 Hz, OH), 6.77 (1H, d,
J = 3.5 Hz, Thio), 7.14 (1H, d, J = 8.0 Hz, Ph), 7.20 (2H, quasi-t,
J = 8.8 Hz, Ph), 7.26 (1H, d, J = 3.5 Hz, thio), 7.32 (1H, dd, J =
8.0, 1.5 Hz, Ph), 7.42 (1H, d, J = 1.5 Hz, Ph), 7.57 (2H, m, Ph).
A solution of the above obtained 1-(1-methoxyglucopy-
ranosyl)-4-methyl-3-(5-(4-fluorophenyl)-2-thienylmethyl)benzene
6b-3 (63.1 g, 132 mmol) and triethylsilane (46.4 g, 399 mmol) in
dichloromethane (660 mL) was cooled in a dry ice-acetone bath
under argon atmosphere, and to the mixture was added dropwise
boron trifluoride-ethyl ether complex (50.0 mL, 395 mmol) over
5 min. The mixture was stirred at the same temperature. The mix-
ture was allowed to warm to 0 °C and stirred under ice-water for
2 h. At the same temperature, a saturated aqueous sodium hydro-
gen carbonate solution (800 mL) was added, and the mixture was
stirred for 30 min. The organic solvent was evaporated under
reduced pressure, and the residue was poured into water (1,500
mL) and extracted with ethyl acetate (1,000 mL) twice. The com-
bined organic layer was washed with water (500 mL) twice, dried
over magnesium sulfate, and treated with activated carbon. The
insoluble was filtered off and the solvent was evaporated under
reduced pressure. The residue was crystallized from ethyl acetate
(300 mL)-diethyl ether (600 mL)-water (6 mL) to give desired
1-(β-D-glucopyranosyl)-4-methyl-3-(5-(4-fluorophenyl)-2-thienyl-
methyl)benzene 4b-3 (33.5 g, 56.7%) as colorless crystals:
mp 98-100 °C. IR (Nujol) 1626, 1600, 1549, 1507 cm-1. HPLC
99.5% (tR = 11.6 min, L-column ODS (5 μm particle size, 4.6 ꢀ
150 mm), CH3CN/20 mM phosphate buffer (pH 6.5) (40/60)).
APCI-MS m/z 462 (M þ NH4). 1H NMR (DMSO-d6) δ 2.26 (3H,
s, Me), 3.13-3.28 (4H, m, sugar), 3.44 (1H, m, sugar), 3.69 (1H, m,
sugar), 3.96 (1H, d, J = 9.3 Hz, sugar), 4.10, 4.15 (each 1H, d, J =
16.0 Hz, Ph-CH2-thio), 4.43 (1H, t, J = 5.8 Hz, OH), 4.72 (1H, d,
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