Synthesis and biological evaluation of novel C-aryl D-glucofuranosides as sodium-dependent glucose co-transporter 2 inhibitors

Article abstract of DOI:10.1016/j.bmc.2013.08.067

Novel C-aryl-d-glucofuranosides were synthesized and evaluated for their capacity to inhibit human sodium-dependent glucose co-transporter 2 (hSGLT2) and hSGLT1. Compound 21q demonstrated the best in vitro inhibitory activity against SGLT2 in this series (EC₅₀ = 0.62 μM).

Full text of DOI:10.1016/j.bmc.2013.08.067

Bioorganic & Medicinal Chemistry 21 (2013) 6282–6291
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Synthesis and biological evaluation of novel C-aryl D-glucofurano-
sides as sodium-dependent glucose co-transporter 2 inhibitors
Tzung-Sheng Lin a, Ya-Wen Liw a, Jen-Shin Song b, Tsung-Chih Hsieh b, Hsien-Wei Yeh a, Lih-Ching Hsu a,
Chun-Jung Lin a, Szu-Huei Wu b, Pi-Hui Liang a,
⇑
a School of Pharmacy, College of Medicine, National Taiwan University, Taipei 100, Taiwan
b Institute of Biotechnology and Pharmaceutical Research, National Health Research Institute, Miaoli County 35053, Taiwan
a r t i c l e i n f o
a b s t r a c t
Article history:
Novel C-aryl-
sodium-dependent glucose co-transporter 2 (hSGLT2) and hSGLT1. Compound 21q demonstrated the
best in vitro inhibitory activity against SGLT2 in this series (EC50 = 0.62 M).
Ó 2013 Elsevier Ltd. All rights reserved.
D-glucofuranosides were synthesized and evaluated for their capacity to inhibit human
Received 29 July 2013
Revised 28 August 2013
Accepted 29 August 2013
Available online 8 September 2013
l
Keywords:
Antidiabetics
SGLT
Glucofuranosides
1. Introduction
and the alternative glycones, for example L
-xyloside,21N-xylo-
sides,22,23 5-thioglucosides,24O-spiroketal glucosides25–28 have
been synthesized and tested in the pursuit novel compounds with
increased efficacy, decrease toxicity (an increase of cancer risks was
reported in the late clinical stage of Dapagliflozin),29 and com-
pounds that also avoid infringement of exiting patents. Several of
these are currently in clinical trials for the treatment of type II dia-
betics, for example Canagliflozin,15 Empagliflozin,19 LX4211,23
ASP1941,18 TS-071,24 PF-04971729,26 Tofogliflozin,27 and YM543.20
Glucose exists predominantly (>99%) in the pyranose form in
aqueous solution; the furanose form exists only in negligible
amounts.30 Accordingly, glucopyranose is most likely to be trans-
ported by SGLT2. To date, and to the best of our knowledge, there
are no reports of a furanose-type five membered-ring being trans-
ported by a glucose-transporter. In the present study, we designed
Phlorizin (1, Fig. 1), a glucose derivative extracted from the root
bark of the apple tree, is found to induce glycosuria through the
inhibition of the sodium dependent glucose cotransporters SGLT1
& SGLT2 in the renal tubules.1–3 SGLT1 is distributed mainly in
the small intestine and SGLT2 is mainly in the proximal tubule of
kidneys. People with SGLT1 gene mutation are unable to transport
glucose or galactose through the intestine wall, resulting in symp-
tom of glucose–galactose malaborption.4,5In contrast, people with
gene mutation in SGLT2 exhibit persistent renal glucosuria, but
are otherwise healthy.6,7 Recent research results indicate that the
plasma glucose level of diabetic patients can be lowered by selec-
tive inhibition of SGLT2.8,9 Phorizin (1) has been used to study
the mechanism of action of renal glucose transporters, but was
not developed into a drug because of its poor intestinal absorption
and tendency to hydrolyze at the glycosidic link at the physiological
conditions. Recently, phlorizin has been derivatized to produce the
1st generation of O-glucoside SGLT2 inhibitors;10–13C-glucoside
analogues were also synthesized to address the problem of hydro-
lysis. The C-glucoside—dapagliflozin (2, Fig. 1) was approved in the
European Union (EU) in November of 2012, as the first drug in the
SGLT2 class for type II diabetics.14 Dapagliflozin is a potent and
selective SGLT2 inhibitor, and can bind to but not be transported
by SGLT2. Since the disclosure of Dapagliflozin, modified C-gluco-
sides bearing a heteroaromatic ring on the aglycon moiety15–20
novel
SGLT inhibitions. Inspired by dapagliflozin, the aglycone of new
-glucofuranoside derivatives consisted a diphenylmethyl group
D-glucofuranoside derivatives to evaluate their activities on
D
shown as compound 3 (Fig. 1). Compounds bearing different sub-
stitutions (compound 3 in Fig. 1) on the diphenylmethyl group
were also explored.
2. Results and discussion
2.1. Chemistry
To generate protected furanose for coupling reaction with a
lithiated diphenylmethyl group, 5,6-O-isopropylidene-D-glucono-
⇑
Corresponding author. Tel.: +886 233668695.
E-mail address: phliang@ntu.edu.tw (P.-H. Liang).
0968-0896/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmc.2013.08.067

T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
6283
soft nucleophile (EtSH) system33 to form the desired products 20a–
20h. Since the methylene group of 19i was not stable under the
hard acid condition, hydrogenation of 19i with Pd/C yielded 20i.
It was found that to achieve complete debenzylation while avoid-
ing de-chlorination and the formation of side products with an
open ring at the C-1 position, the reaction time must be restricted
within 16–20 h.
Compounds 27j–27q were synthesized in a manner similar to
that reported previously,24 with modifications. Commercially
available benzoic acids (21 and 22, Scheme 3) were converted to
the corresponding benzoyl chlorides with oxalyl chloride and un-
der Friedel–Crafts conditions, were acylated with phenyl group
(23–25) to generate a 7:1 mixture of regioisomers in favor of the
desired p-benzophenone 26j–26n, which were purified by recrys-
tallization from ethanol. Reduction of 26j–26n with BF3OEt2 and
Et3SiH yielded aglycone 27j–27n. Compounds 27o–27q were also
synthesized (Scheme 3). After methylation of commercially avail-
able 28 with iodomethane under basic conditions, Friedel-Crafts
reaction was performed using benzoyl chloride (29–31) to afford
26o–26q. Subsequent reduction of 26o–26q using the same condi-
tions described above gave the aglycon portions 27o–27q.
Lithium–halogen exchange, followed by the addition of the nas-
cent lithiated aromatic compound to per-benzylated 11, produced
a mixture of the corresponding lactols (32j–32q, Scheme 4). Reduc-
tion of the anomeric hydroxy group with BF3OEt2 and Et3SiH gave
OH
OH
Cl
OEt
HO
OH
OH
O
O
HO
HO
O
OH
HO
HO
O
Dapagliflozin (2)
OH
phlorizin (1)
HO
HO
OH
O
O
HO
HO
OH
OH
OH
OH
HO
R3
HO
HO
R1
R2
O
HO
OH
3
Figure 1. Structures of SGLT2 inhibitors.
1,4-lactone 5 was prepared from gluconolactone 4 as depicted in
Scheme 1.31 Silylated lactone 6 was obtained by treating 5 with
TMSCl and NMM. Nucleophilic addition of compounds 7 to 6 was
accomplished by lithium–halogen exchange using n-BuLi in THF
and toluene; the resulting hydroxyl group was trapped by MeOH
under MsOH promotion. This reaction was expected to yield the
corresponding O-methyl lactol 8, but instead, compound 9 was ob-
tained, presumably by acetonide hydrolysis (caused by addition of
MsOH) followed by opening of the five-membered ring and closure
of the six-membered ring.
33j–33q as a mixture of anomers (b/a ratio about 4:1). It was found
that 33o–33q could be separated in their perbenzylated forms by
column chromatography on silica gel, but separation of 33j–33n
required their conversion to the corresponding tetra-acetates
(Scheme 4). Debenzylation of perbenzylated C-glucoside were car-
ried out in the hard acid (BF3) and soft nucleophile (EtSH) system33
to obtain 20j–20q.
To overcome this problem, perbenzylated glucono-
c-lactone 11
(synthesized by PCC oxidation of the per-benzyl furanoside 1032
)
was used instead of compound 6 (Scheme 2). The commercially
available substituted benzoic acid 12 was converted to the corre-
sponding methyl benzoate, which was then reduced with LiAlH4
to form a benzyl alcohol and subsequently silylation with TIPSCl
in the presence of imidazole and DMAP to generate 13.16 Lith-
ium–halogen exchanged, followed by the addition of the nascent
2.2. Biological evaluation
Biological evaluation of 20a–20q could now take place; the re-
sults are presented in Table 1. EC50 values were calculated by mea-
suring inhibition of the sodium-dependent uptake of [14C]-labeled
lithiated aromatic compound to perbenzylated glucono-c-lactone
a-methyl-D-glucopyranoside (AMG) into Chinese hamster ovary
11, produced a mixture of corresponding lactols 14. These were re-
duced using BF3OEt2 and Et3SiH to give compound 15, which was
desilylated using TBAF and finally purified by column chromatog-
raphy on silica gel to afford alcohol 16 in a single b-anomer at
44% yield over three steps. Alcohol 16 was oxidized by PCC to form
aldehyde 17 in 87% yield. Addition of various Grignard reagents to
17 to introduce the substituted, heteroaryl and benzofused rings at
the distal ring position. The resulting alcohols 18a–18i were re-
duced under standard conditions (BF3OEt2 and Et3SiH) to generate
19a–19i. 19a–19h were debenzylated by using hard acid (BF3) and
(CHO) cells stably expressing human SGLT2 (hSGLT2) or
hSGLT1.34,35 Because differences in protein expression levels of
the cells will result in differences of EC50 values, the SGLT inhibitor
phlorizin (1) was used as a reference compound in this in vitro
activity evaluation system.
Inspired by dapagliflozin, the diphenyl methyl moiety (20a) was
established as a standard and substitution at the central and distal
rings was investigated; previous reports suggest the position of
substitution to be important.18,23,28 Firstly, the chlorine group at
R2 was retained and the substituents on the distal phenyl ring
O
O
Cl
OEt
O
d
d
O
O
O
O
OMe
HO
HO
8
OH
O
O
O
O
O
a
b
HO
HO
O
HO
HO
OH
TMSO
OTMS
HO
4
Cl
OEt
6
5
O
HO
HO
HO
Cl
OEt
OMe
9
c
Br
7
Scheme 1. Reagents and conditions: (a) SnCl2, 2,2-DMP, DMF, 40 °C; (b) TMSCl, NMM, THF, 0–24 °C, 20 h; (c) n-BuLi, THF, toluene, À78 °C; (d) (i) toluene, À78 °C; (ii) MsOH,
MeOH, À78 °C to rt.

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T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
BnO
BnO
BnO
BnO
BnO
BnO
Cl
BnO
BnO
Cl
a
b
d
O
O
e
OH
OBn
O
O
O
OTIPS
O
OTIPS
OH
OBn
OBn
BnO
BnO
11
BnO
OBn
BnO
c
10
15
14
f
Cl
Cl
BnO
BnO
Cl
H
BnO
BnO
Cl
g
OTIPS
O
O
Br
CO2H
OH
Cl
Br
13
12
BnO
OBn
BnO
OBn
17
16
h
BnO
BnO
BnO
BnO
HO
HO
Cl
Cl
j
i
O
O
O
R
R
R
OH
BnO
BnO
HO
OBn
18a-18i
OBn
19a-19i
OH
20a-20h
k
O
g, R =
h, R =
a, R =4-OMe-Ph
b, R =3-OMe-Ph
c, R =2-OMe-Ph
d, R =4-tBu-Ph
e, R =4-F-Ph
OMe
20i
f, R =4-SMe-Ph
O
O
i, R =
Scheme 2. Reagents and conditions: preparation of b-C-glucoside 20a–20i. (a) PCC, CH2Cl2, rt, 2 h, 92%; (b) (i) MeI, K2CO3, DMF, rt, 3 h, 98%; (ii) LiAlH4, Tol, rt, 2 h, 92%; (iii)
TIPSCl, imidazole, DMAP, DMF, rt, 81%; (c) n-BuLi, THF/Tol, À78 °C, 1 h; (d) THF, À78 °C, 3 h; (e) BF3OEt2, Et3SiH, CH2Cl2, À50 to À10 °C, 1 h; (f) TBAF, THF, rt, 2 h, 44% (3 steps).
(g) PCC, CH2Cl2, rt, 2 h, 87%; (h) Grignard reagents, THF, À78 °C, 3 h; (i) BF3OEt2, Et3SiH, CH2Cl2, 0 °C, 1 h, 19–83%, two steps; (g) BF3OEt2, EtSH, CH2Cl2, rt, 20 h, 15–50%; (k) Pd/
C, H2, CH2Cl2/MeOH (3:1), 20 h, 35%.
R1
Br
R2
R3
R1
Br
R2
R3
R1
Br
R2
O
b
c
a
OH
O
27j-27n
R3
21 R1=H, R2=Cl
22
26j-26n
j, R1 =H, R2 =Cl, R3 =OEt
R1=H, R2=Me
23, R3 =OEt
, R3 =iPr
, R3 =Et
k
, R1 =H, R2 =Cl, R3 =iPr
24
25
l
,
R1 =H, R2 =Me, R3 =OEt
, R1 =H, R2 =Me, R3 =iPr
m
n, R1 =H, R2 =Me, R3 =Et
R1
Br
R2
O
R3
R1
Br
28 R1=OH, R2=Cl
R2
d
e
c
R1
Br
R2
R3
R3
Cl
27o-27q
O
29
26o-26q
o
p
, R1 =OMe, R2 =Cl, R3 =OEt
, R3 =OEt
30, R3 =iPr
31, R3 =Et
, R1 =OMe, R2 =Cl, R3 =iPr
q, R1 =OMe, R2 =Cl, R3 =Et
Scheme 3. Reagents and conditions: synthesis of aglycon 27j–27q. (a) (COCl)2, DMF, CH2Cl2,20 h, rt; (b) AlCl3, CH2Cl2, 3 h, 0–24 °C; (c) BF3OEt2, Et3SiH, CH2Cl2,1 h, 0–24 °C; (d)
MeI, K2CO3, DMF, 5 h, rt; (e) AlCl3, CH2Cl2, 20 h, 0–40 °C.
varied. It was found that only molecules bearing para-substituents
imparted inhibitory activity (20a, EC50 of 35 M for SGLT2). The
introduction of tert-butyl (20d), methylthio (20f), and isopropyl
groups (20k) enhanced the SGLT2 inhibition potency; the ethoxyl
(20j), fluoro (20e), and phenyloxyl-bearing (20g) analogues had
no inhibition activity. Compound 20f bearing a methylthio moiety
SGLT2 of 5.1 lM, superior to that of 20a. However, introduction
l
of a 3.4-methylenedioxybenzyl group (20i) resulted in no inhibi-
tion. This suggests that the presence of heteroaryl groups or groups
bearing bulky substitutions on the distal phenyl ring improve po-
tency, but the effect is limited to certain heteroaryl groups.
We next studied the effect of substitution on the central ben-
zene moiety. Introduction of methyl group at R2 (20l, 20m, and
20n) slightly increased the SGLT2 inhibition potency compared
to 20j and 20k. The presence of an ethyl group at the para-position
of the distal phenyl ring resulted in increased potency (EC50 = 2.5 -
had an EC50 of 3.7 lM and selectivity for SGLT2 of over 13-fold. It
was found that the substituent at the para position of the distal
ring could occupy a two- or three-carbon-sized pocket of hSGLT2.
On the other hand, inhibitors bearing relatively hydrophilic sub-
stituents such as 20a (R3 = OMe) and 21j (R3 = OEt) exhibited less
potent inhibition, compared analogues bearing hydrophobic alkyl
substituents. The introduction of a fluoro group (20e) resulted in
a loss of potency. Replacement of the phenyl group with the ben-
zofused ring naphthalene gave compound 20h with an EC50 for
l
M) and selectivity (>20-fold). Introduction of methoxy group at
R1 has been reported to increase selectivity,18,24 compounds 20o,
20p, and 20q were thus synthesized. Indeed, compound 20q
turned out to be the most potent inhibitor with an EC50 of
0.62 lM of SGLT2, and 47-fold selectivity against SGLT1. As

T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
6285
R1
Br
R2
R3
BnO
BnO
BnO
BnO
R1
R2
R3
b
27j-27q
O
O
O
a
OH
OBn
OBn
BnO
BnO
32j-32q
11
BnO
BnO
HO
HO
R1
R2
R3
R1
R2
R3
c
O
O
BnO
OBn
HO
OH
20j-20q
33j-33q
n, R1 =H, R2 =Me, R3 =Et
o, R1 =OMe, R2 =Cl, R3 =OEt
p, R1 =OMe, R2 =Cl, R3 =iPr
q, R1 =OMe, R2 =Cl, R3 =Et
j, R1 =H, R2 =Cl, R3 =OEt
k, R1 =H, R2 =Cl, R3 =iPr
l, R1 =H, R2 =Me, R3 =OEt
m, R1 =H, R2 =Me, R3 =iPr
Scheme 4. Reagents and conditions: synthesis of compounds 20j–20q. (a) 27j–27q Treated with n-BuLi, THF/Toluene, À78 °C, 1 h and then 11, THF, À78 °C, 3 h; (b) BF3OEt2,
Et3SiH, CH2Cl2, 0–24 °C, 1 h; (c) (i) BF3OEt2, EtSH, CH2Cl2, rt, 20 h; (ii) Ac2O, pyridine, DMAP, rt, 2 h, for compounds 20j–20n; (iii) NaOMe, MeOH, rt, 3 h for compounds 20o–
20q.
Table 1
In vitro data for hSGLT inhibitory activity and selectivitya
HO
HO
R1
R2
O
R3
HO
OH
Compds
R1
R2
R3
hSGLT2 EC50
(l
M)
hSGLT1 EC50
(
lM)
Selectivity (T1/T2)
20a
20b
20c
20d
20e
20f
20g
20h
20i
20j
20k
20l
20m
20n
20o
20p
20q
1
H
H
H
H
H
H
H
H
H
H
H
H
H
H
OMe
OMe
OMe
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Me
Me
Me
Cl
Cl
Cl
4-OMe–Ph
3-OMe–Ph
2-OMe–Ph
4-tBu–Ph
35
>50
NDb
ND
24
ND
>50
ND
>50
ND
ND
47
>50
>50
>50
>50
>50
29
>1.4
—
—
1
—
>13.3
—
>9.7
—
>50
>50
24
>50
3.7
>50
5.1
>50
>50
33
34
11
2.5
46
21
4-F–Ph
4-SMe-Ph
4-Ph–O–Ph
4-(6-OMe)-Naph
4-(3,4-Methylenedioxy)–Ph
4-OEt–Ph
4-iPr–Ph
4-OEt–Ph
4-iPr–Ph
4-Et–Ph
—
2.2
>1.5
>20
>4.5
>1
>2.4
47
1.6
4-OEt–Ph
4-iPr–Ph
4-Et–Ph
0.62
0.077
0.12
a
EC50 values for SGLT1 and SGLT 2 activities represent the mean of at least three determinations.
ND: not determined.
b
previously described in the synthesis of TS-071 and ASP1941, the
methoxy group at R1 results in greater selectivity for SGLT2, and
the methyl or chloro group at R2 is responsible for the increased
potency than phlorizin, most had higher selectivity for SGLT2 com-
pared to phlorizin.
potency towards SGLT218,24
D
-Glucofuranosides inhibitors devel-
oped by us have similar SAR results with previous studies using
C-
-glucopyranose,18 or 1-thio- -glucitol,24 indicating that the
binding mode of -glucofuranosides compared with -glucopyra-
3. Conclusion
D
D
In the present study, the synthesis of C-aryl D-glucofuranosides
D
D
20a–20p has been accomplished and SAR studies relating to the
substitution of various groups on the aromatic rings and their ef-
fect on SGLT inhibition conducted. It was found that a methoxy
group adjacent to the sugar moiety was important for SGLT2 selec-
tivity, whereas a chloro or methyl group para to the sugar moiety
increased SGLT2 potency. Of the derivatives tested, compound
20q was found to be the most potent inhibitor with EC50 of
0.62 lM of SGLT2 and a 47-fold selectivity against SGLT1. Although
this series of compounds failed to advance as a development can-
didate, the chemistry shown here promoted us to synthesis more
nose might be similar. C-aryl glucofuranosides (e.g. compound
20q) are more than 100-fold weaker than C-aryl glucopyranosides
(e.g. dapagliflozin) in the SGLT2 inhibition, which can possibly be
explained by that SGLT2 recognizes glucopyranose instead of
glucofuranose. Since the pyranose form of glucose in the aqueous
solution exists more than 99% and the furanose form exists less
than 1%,30 our results indirectly prove that SGLT2 primarily
transports the pyranose type of glucose. Although none of C-aryl
D
-glucofuranosides synthesized herein exhibited higher inhibitory

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T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
diversified, yet desirable targets in a relatively straightforward
fashion. Further modification of this series will be reported in
due course.
(d, J = 2.6, 1H), 4.94 (d, J = 11.3 Hz, 1H), 4.79–4.56 (m, 7H), 4.54–
4.39 (m, 3H), 4.37–4.31 (m, 1H), 4.27 (d, J = 3.2 Hz, 1H), 4.13–
4.02 (m, 2H), 3.87 (dd, J = 5.6, 10.6 Hz, 1H); 13C NMR (100 MHz,
CDCl3) d 139.7, 138.7, 138.3, 138.0, 137.5, 137.3, 131.3, 128.9,
128.3, 128.1, 128.1, 127.7, 127.5, 127.5, 127.4, 127.4, 127.3,
126.6, 126.5, 88.4, 85.9, 82.3, 80.8, 75.8, 73.3, 72.5, 71.6, 71.2,
4. Experimental section
4.1. Chemistry
71.0, 62.4; HRMS (ESI) calcd for
687.2484, found 687.2492.
C
41H41ClO6Na+ [M+Na]+
Reactions were monitored by thin-layer chromatography
(Merck, silica gel 60F-254) using p-anisaldehyde or cerium molyb-
date as the stain reagent. Silica gel used for flash column chroma-
tography was Mallinckrodt type 60 (230–400 mesh). Unless
otherwise noted, reagents and materials were obtained from com-
mercial sources and used as provided without further purification.
1H and 13C NMR spectra were recorded on a Bruker AV-400 or AVI-
II-600-Cry spectrometer and referenced to residual solvent peaks
(CDCl3: 1H d = 7.24, 13C d = 77.0; CD3OD: 1H d = 3.30, 13C d = 49). Ex-
act mass measurements were performed on VG plateform electro-
spray ESI/MS or BioTOF II (Taiwan).
4.1.3. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-(4-chloro-3-
oxomethylphenyl)- -glucitol (17)
D
To a solution of compound 16 (4.0 g, 6.01 mmol) in dry CH2Cl2
(500 mL) was added freshly activated 4 Å molecular sieves (6 g)
and PCC (3.24 g, 15.0 mmol). The reaction mixture was stirred at
room temperature. After 5 h, the mixture was concentrated in va-
cuo and the residue was filtered through silica gel (10 g). The filter
cake was washed with CH2Cl2. The combined filtrate were concen-
trated in vacuo to afford compound 17 (3.47 g, 5.23 mmol, 87%
yield) as a colorless oil. 1H NMR (400 MHz, CDCl3) d 10.47 (s,
1H), 7.89–7.88 (d, 1H), 7.54 (dd, J = 8.4, 1.6 Hz, 1H), 7.36–7.23
(m, 19H), 7.06–6.87 (m, 2H), 4.96–4.83 (m, 2H), 4.63–4.50 (m,
5H), 4.38 (s, 1H), 4.34 (dd, J = 9.1, 3.2 Hz, 1H), 4.27–4.22 (m, 1H),
4.17–3.91 (m, 4H), 3.79 (dd, J = 10.6, 5.4 Hz, 1H). 13C NMR
(100 MHz, CDCl3) d 189.5, 140.6, 138.7, 138.4, 137.4, 137.2,
136.5, 133.1, 132.0, 130.4, 128.5, 128.4, 128.2, 128.2, 128.2,
127.9, 127.6, 127.6, 127.5, 127.5 127.4, 127.4, 127.1, 88.2, 85.3,
82.2, 81.1, 75.8, 73.4, 72.6, 71.8, 71.4, 71.0; HRMS (ESI) calcd for
4.1.1. 2,3,5,6-O-Tetrabenzyl-glucono-gamma-lactone (11)
To a solution of compound 1032(52.0 g, 96.2 mmol) in dry
CH2Cl2 (500 mL) was added freshly activated 4 Å molecular
sieves (60 g) and PCC (36.0 g, 167 mmol). The reaction mixture
was stirred at room temperature. After 5 h, the mixture was con-
centrated in vacuo and the residue was filtered through silica gel
(50 g). The filter cake was washed with CH2Cl2. The combined
filtrate were concentrated in vacuo to afford compound 11
C
41H39ClO6 [M+Na]+ 685.2327, found 685.2338.
(47.9 g, 88.9 mmol, 92% yield) as
a
colorless oil. 1H NMR
(400 MHz, CDCl3) d 7.43–7.17 (m, 20H), 4.92 (d, J = 11.7 Hz,
1H), 4.80 (dd, J = 6.3, 5.4 Hz, 1H), 4.76 (d, J = 11.2 Hz, 1H), 4.63
(d, J = 11.6 Hz, 1H), 4.60–4.45 (m, 5H), 4.35–4.27 (m, 2H), 4.14–
4.07 (m, 1H), 3.87–3.71 (m, 2H).13C NMR (50 MHz, CDCl3) d
172.8, 137.9, 137.9, 136.9, 136.6, 128.4, 128.3, 128.3, 128.2,
128.1, 128.0, 127.8, 127.6, 127.5, 127.5, 79.1, 78.7, 76.7, 76.3,
73.3, 72.9, 72.3, 72.2, 69.5; HRMS (ESI) calcd for C34H34O6Na+
[M+Na]+ 561.2253, found 561.2260.
4.1.4. General procedure for the synthesis of compound 19a.
(1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-(4-
methoxybenzyl)-phenyl]-D-glucitol (19a)
To a solution of compound 17 (150 mg, 0.23 mmol) in anhy-
drous THF (2 mL) under N2 atmosphere was added 4-methoxyphe-
nylmagnesium bromide (1.60 mL, 0.80 mmol, 0.5 M in THF)
dropwise at À78 °C. After stirring for 2.5 h at À78 °C, the reaction
was quenched by the addition of a saturated aqueous solution of
NH4Cl (5 mL) and the resulting mixture was extracted with CH2Cl2.
The organic phase was removed under reduced pressure and co-
evaporated with toluene twice to afford crude 18a. A solution of
the crude 18a in CH2Cl2 (5 mL) was cooled to À50 °C, and Et3SiH
4.1.2. General procedures for synthesis of 16. (1S)-1,4-Anhydro-
2,3,5,6-tetra-O-benzyl-1-C-[(4-chloro-3-hydroxymethyl)-
phenyl]-D-glucitol (16)
To a solution of compound 1316 (6.44 g, 17.0 mmol) in THF/tol-
uene (1:2, 30 mL) under N2 gas was added n-BuLi (2.5 M in hexane,
6.8 mL, 17.0 mmol) dropwise at À78 °C. After 1 h, this mixture was
added to a solution of 11 (7.35 g, 13.6 mmol) in toluene (10 mL),
while maintaining the temperature at À78 °C. After 3 h, the mix-
ture was quenched with NaHCO3 solution and extracted with CH2-
Cl2. The organic layer was then washed with H2O, brine, dried over
MgSO4, filtered and concentrated under reduced pressure to afford
crude 14. Compound 14 was dissolved in CH2Cl2 (10 mL) and the
solution cooled to À50 °C. To this solution was added Et3SiH
(4 mL, 25.0 mmol), followed by BF3OEt2 (3.2 mL, 27.5 mmol)
slowly to maintain the temperature below À40 °C. The resulting
suspension was allowed to warm to À10 °C over 1 h prior to
quenching with K2CO3. After concentration in vacuo, the resulting
mixture was extracted with EtOAc and washed with brine, dried
over MgSO4, filtered and concentrated to afford crude 15 as yellow
oil. Then, compound 15 in THF (10 mL) was added TBAF (1.0 M in
THF, 17.0 mL, 17.0 mmol) and the reaction mixture was stirred at
ambient temperature for 2 h. After concentration in vacuo, the res-
idue was extracted twice between EtOAc and NH4Cl. The combined
organic layers were washed with brine, dried over MgSO4, filtered
and purified with column chromatography (200 g silica gel, EtOAc/
toluene/n-hexane = 1:1:8) to afford pure beta form of 16 (4.0 g,
6.01 mmol, 44% yield over three steps) as a colorless oil. 1H NMR
(400 MHz, CDCl3) d 7.59–7.28 (m, 21H), 7.25–7.14 (m, 2H), 5.01
(53 lL, 0.33 mmol) added, followed by BF3OEt2 (26 lL, 0.22 mmol)
slowly; the temperature was maintained below À40 °C. The result-
ing suspension was allowed to warm to À10 °C over 1 h prior to
quenching with K2CO3. After concentration in vacuo, the resulting
mixture was extracted with EtOAc and washed with brine, dried
over MgSO4, filtered and purified with column chromatography
(5 g silica gel, EtOAc/n-hexane = 1/20) to afford 19a (138 mg,
0.18 mmol, 79% yield over two steps) as colorless oil. 1H NMR
(400 MHz, CDCl3) d 7.49–7.29 (m, 18H), 7.28–7.22 (m, 3H), 7.15–
7.08 (m, 4H), 6.89–6.83 (m, 2H), 4.97 (d, J = 2.6 Hz, 1H), 4.93 (d,
J = 11.4 Hz, 1H), 4.69 (d, J = 2.1 Hz, 2H), 4.64 (d, J = 11.6 Hz, 1H),
4.62–4.52 (m, 2H), 4.45 (d, J = 3.0 Hz, 2H), 4.41 (dd, J = 9.0, 3.4 Hz,
1H), 4.27–4.21 (m, 2H), 4.09.4.00 (m, 4H), 3.83 (dd, J = 10.6,
5.6 Hz, 1H), 3.78 (s, 3H); 13C NMR (100 MHz, CDCl3) d 157.9,
139.6, 138.8, 138.7, 138.5, 137.6, 137.4, 132.8, 131.3, 129.8,
129.1, 128.8, 128.3, 128.1, 128.1, 127.7, 127.5, 127.4, 127.4,
127.3, 127.2, 125.4, 113.7, 88.4, 85.7, 82.4, 80.8, 75.9, 73.3, 72.4,
71.6, 71.2, 70.9, 54.9, 38.2; HRMS (ESI) calcd for C48H47ClO6
[M+Na]+ 777.2953, found 777.2903.
4.1.5. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-
(3-methoxybenzyl)-phenyl]-D-glucitol (19b)
Compound 19b (33 mg, 0.044 mmol, 19% yield over two steps)
was prepared as a colorless oil from 17 (150 mg, 0.23 mmol) and
3-methoxyphenylmagnesium bromide (0.80 mL, 0.80 mmol,

T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
6287
1.0 M in THF) according to the procedure for the synthesis of 19a.
1H NMR (400 MHz, CDCl3) d 7.38–7.11 (m, 22H), 7.04–6.98 (m, 2H),
6.74–6.67 (m, 3H), 4.84 (d, J = 2.6 Hz, 1H), 4.81 (d, J = 11.4 Hz, 1H),
4.63–4.42 (m, 5H), 4.39–4.30 (m, 2H), 4.26 (dd, J = 9.1, 3.4 Hz, 1H),
4.15–4.09 (m, 2H), 4.01 (s, 2H), 3.95–3.88 (m, 2H), 3.72 (dd,
J = 11.0, 5.2 Hz, 1H), 3.69 (s, 3H); 13C NMR (100 MHz, CDCl3) d
159.6, 141.0, 139.7, 138.9, 138.6, 138.1, 137.6, 137.5, 133.0,
129.3, 129.0, 128.4, 128.2, 128.2, 127.8, 127.6, 127.6, 127.5,
127.5, 127.3, 125.7, 121.3, 114.8, 111.4, 88.5, 85.8, 82.4, 80.9,
76.0, 73.4, 72.6, 71.7, 71.3, 71.1, 55.0, 39.1; HRMS [ESI] calcd for
4.1.9. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-
(4-methylthiobenzyl)-phenyl]- -glucitol (19f)
D
Compound 19f (102 mg, 0.13 mmol, 57% yield, two steps) was
prepared as a colorless oil from 17 (150 mg, 0.23 mmol) and 4-
thioanisolemagnesium bromide (1.60 mL, 0.80 mmol, 0.5 M in
THF), according to the procedure for the synthesis of 19a. 1H
NMR (400 MHz, CDCl3) d 7.44–7.27 (m, 19H), 7.25–7.15 (m, 4H),
7.10–7.05 (m, 4H), 4.94 (d, J = 2.5 Hz, 1H), 4.90 (d, J = 11.5 Hz,
1H), 4.66 (d, J = 1.5 Hz, 2H), 4.63–4.54 (m, 3H), 4.41 (d, J = 2.4 Hz,
2H), 4.37 (dd, J = 9.0, 3.3 Hz, 1H), 4.22–4.16 (m, 2H), 4.04 (s, 2H),
4.01 (dd, J = 10.7, 1.9 Hz, 1H), 3.98 (d, J = 2.5 Hz, 1H), 3.78 (dd,
J = 10.7, 5.5 Hz, 1H), 2.44 (s, 3H); 13C NMR (100 MHz, CDCl3) d
139.7, 138.8, 138.5, 138.1, 137.5, 137.4, 136.2, 135.8, 132.8,
129.3, 129.2, 128.9, 128.4, 128.1, 127.8, 127.5, 127.5, 127.4,
127.4, 127.3, 126.7, 125.6, 88.4, 85.7, 82.3, 80.8, 75.9, 73.3, 72.4,
71.6, 71.2, 70.8, 38.5, 15.8; HRMS (ESI) calcd for C48H47ClO5S
[M+H]+ 771.2905, found 771.2879.
C
48H47ClO6 [M+Na]+ 777.2953, found 777.2914.
4.1.6. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-
(2-methoxybenzyl)-phenyl]- -glucitol (19c)
D
Compound 19c (145 mg, 0.19 mmol, 83% yield over two steps)
was prepared as a colorless oil from 17 (150 mg, 0.23 mmol) and
2-methoxyphenylmagnesium bromide (0.80 mL, 0.80 mmol,
1.0 M in THF) according to the procedure for the synthesis of
19a. 1H NMR (400 MHz, CDCl3) d 7.50–7.23 (m, 22H), 7.18–7.10
(m, 2H), 7.07–7.01 (m, 1H), 6.97–689 (m, 2H), 4.97 (d, J = 2.7 Hz,
1H), 4.94 (d, J = 11.6 Hz, 1H), 4.74–4.65 (m, 2H), 4.65 (d,
J = 11.6 Hz, 1H), 4.58 (s, 2H), 4.50–4.42 (m, 2H), 4.40 (dd, J = 9.1,
3.4 Hz, 1H), 4.28–4.23 (m, 2H), 4.18 (s, 2H), 4.07–4.01 (m, 2H),
3.87–3.80 (m, 4H); 13C NMR (100 MHz, CDCl3) d 157.3, 139.4,
138.8, 138.5, 137.9, 137.5, 137.4, 133.1, 130.0, 129.0, 128.9,
128.3, 128.1, 128.1, 127.7, 127.6, 127.5, 127.4, 127.3, 125.3,
120.3, 110.1, 88.5, 85.8, 82.4, 80.8, 75.8, 73.3, 72.5, 71.5, 71.2,
4.1.10. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-
3-(4-phenoxybenzyl)-phenyl]-D-glucitol (19g)
Compound 19g (131 mg, 0.16 mmol, 70% yield over two steps)
was prepared as a yellowish oil from 17 (150 mg, 0.23 mmol)
and 4-phenoxyphenylmagnesium bromide (1.6 mL, 0.80 mmol,
0.5 M in THF), according to the procedure for the synthesis of
19a. 1H NMR (400 MHz, CDCl3) d 4.48–7.27 (m, 21 H), 7.20–7.04
(m, 9H), 7.00–6.95 (m, 2H), 5.00 (d, J = 2.4 Hz, 1H), 4.95 (d,
J = 11.4 Hz, 1H), 4.72–4.55 (m, 5H), 4.49–4.37 (m, 3H), 4.31–4.21
(m, 2H), 4.11 (s, 2H), 4.08 (dd, J = 10.7, 1.9 Hz, 1H), 4.04 (d,
J = 2.7 Hz, 1H), 3.85 (dd, J = 10.6, 5.5 Hz, 1H); 13C NMR (100 MHz,
CDCl3) d 157.2, 157.1, 155.4, 139.7, 138.8, 138.4, 138.3, 137.5,
137.4, 134.2, 132.8, 130.5, 130.0, 129.6, 129.5, 129.2, 128.9,
128.4, 128.1, 128.1, 127.8, 127.58, 127.50, 127.4, 127.3, 125.6,
123.0, 122.9, 118.7, 118.6, 88.4, 85.7, 82.3, 80.8, 75.9, 73.3, 72.5,
71.6, 71.2, 70.9, 38.3; HRMS (ESI) calcd for C53H49ClO6 [M+Na]+
839.3110, found 839.3074.
71.0, 55.1, 33.2; HRMS (ESI) calcd for
777.2953, found 777.2928
C
48H47ClO6 [M+Na]+
4.1.7. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-
(4-fluorobenzyl)-phenyl]- -glucitol (19d)
D
Compound 19d (134 mg, 0.18 mmol, 78% yield, two steps)
was prepared as a colorless oil from 17 (150 mg, 0.23 mmol)
and 4-fluorophenylmagnesium bromide (0.80 mL, 0.80 mmol,
1.0 M in THF), according to the procedure for the synthesis of
19a. 1H NMR (400 MHz, CDCl3) d 7.24–7.19 (m, 20H), 7.17 (d,
J = 2.0 Hz, 1H), 7.10–7.02 (m, 4H), 6.96–6.89 (m, 2H), 4.91 (d,
J = 2.5 Hz, 1H), 4.88 (d, J = 11.4 Hz, 1H), 4.64 (s, 2H), 4.61–4.49
(m, 3H), 4.40 (s, 2H), 4.34 (dd, J = 9.1, 3.4 Hz, 1H), 4.18 (d,
J = 3.4 Hz, 2H), 4.02 (s, 2H), 3.99 (dd, J = 10.8, 2.0 Hz, 1H), 3.96
(d, J = 2.6 Hz, 1H), 3.77 (dd, J = 10.7, 5.5 Hz, 1H); 13C NMR
(100 MHz, CDCl3) d 162.5, 160.1, 139.8, 138.8, 138.5, 138.1,
137.63, 137.62, 137.4, 135.0, 135.0, 132.9, 130.2, 130.2, 129.3,
128.9, 128.4, 128.2, 127.8, 127.6, 127.5, 127.5, 127.5, 127.3,
125.7, 115.2, 114.9, 88.5, 85.7, 82.4, 80.9, 75.9, 73.4, 72.5, 71.7,
4.1.11. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-
3-(6-methoxy-naphtha-len-2-yl)methyl] phenyl]-
D-glucitol
(19h)
Compound 19h (120 mg, 0.15 mmol, 65% yield, two steps) was
prepared as a yellowish oil from 17 (150 mg, 0.23 mmol) and 6-
methoxy-2-naphthylmagnesium
bromide
solution (1.6 mL,
0.80 mmol, 0.5 M in THF) according to the procedure for the syn-
thesis of 19a. 1H NMR (400 MHz, CDCl3) d 7.83–7.45 (m, 5H),
7.37–7.25 (m, 13H), 7.23–7.13 (m, 7H), 7.10–6.91 (m, 4H), 4.91
(m, 1H), 4.70–4.64 (m, 1H), 4.60–4.51 (m, 2H), 4.50–4.43 (m,
2H), 4.37–4.23 (m, 3H), 4.09 (dd, J = 8.8, 3.5 Hz, 1H), 4.00–3.92
(m, 2H), 3.92–3.87 (m, 2H), 3.87–3.82 (m, 3H), 3.74–3.63 (n, 1H),
3.40–3.33 (m, 2H); 13C NMR (100 MHz, CDCl3) d 157.7, 138.9.
138.6, 137.5, 134.0, 132.1, 129.5, 128.5, 128.4, 128.26, 128.22,
127.8, 127.6, 127.5, 127.5, 127.3, 126.9, 126.6, 126.2, 126.1,
126.0, 125.7, 125.5, 118.7, 105.6, 88.4, 85.7, 82.3, 80.9, 76.0, 73.4,
72.5, 72.4, 71.7, 71.3, 57.1, 29.7; HRMS (ESI) calcd for C52H49ClO6
[M+Na]+ 827.3110, found 827.3217.
71.3, 71.0, 38.3; HRMS (ESI) calcd for
743.2940, found 743.2904.
C
47H44ClFO5 [M+H]+
4.1.8. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-
(4-tert-butylbenzyl)-phenyl]- -glucitol (19e)
D
Compound 19e (125 mg, 0.16 mmol, 70% yield over two steps)
was prepared as a colorless oil from 17 (150 mg, 0.23 mmol) and
4-tert-butylphenylmagnesium
bromide (1.60 mL,
0.80 mmol,
0.5 M in THF), according to the procedure for the synthesis of
19a. 1H NMR (400 MHz, CDCl3) d 7.47–7.20 (m, 23H), 7.14–7.06
(m, 4H), 4.92–4.84 (m, 2H), 4.66–4.56 (m, 3H), 4.55–4.47 (m,
2H), 4.45–4.36 (m, 2H), 4.32 (dd, J = 9.0, 3.4 Hz, 1H), 4.25–4.19
(m, 1H), 4.18 (d, J = 3.4 Hz, 1H), 4.06 (s, 2H), 3.99 (dd, J = 10.8,
1.9 Hz, 1H), 3.95 (d, J = 2.8 Hz, 1H), 3.77 (dd, J = 10.6, 5.6 Hz, 1H),
1.31 (s, 9H); 13C NMR (100 MHz, CDCl3) d 148.8, 139.7, 138.8,
138.5, 138.4, 137.6, 137.4, 136.3, 133.0, 129.3, 129.1, 128.4,
128.4, 128.2, 128.2, 128.2, 127.8, 127.6, 127.5, 127.5, 127.3,
125.6, 125.2, 85.5, 85.9, 82.4, 80.8, 75.9, 73.3, 72.6, 71.7, 71.3,
71.1, 38.6, 34.2, 31.3; HRMS (ESI) calcd for C51H53ClO5 [M+Na]+
803.3474, found 803.3442.
4.1.12. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-
3-(3,4-methylenedioxy-benzyl)phenyl]-D-glucitol (19i)
Compound 19i (106 mg, 0.14 mmol, 61% yield, two steps) was
prepared as a colorless oil from 17 (150 mg, 0.23 mmol) and 3,4-
(methylenedioxy)phenylmagnesium bromide (0.8 mL, 0.80 mmol,
1.0 M in THF/toluene = 1/1), according to the procedure for the
synthesis of 19a. 1H NMR (400 MHz, CDCl3) d 7.41–7.20 (m,
19H), 7.20–7.15 (m, 2H), 7.07–7.02 (m, 2H), 6.69 (d, J = 7.9 Hz,
1H), 6.63–6.56 (m, 2H), 5.85 (dd, J = 5.2, 1.5 Hz, 2H), 4.88 (d,
J = 2.6 Hz, 1H), 4.86 (d, J = 11.5 Hz, 1H), 4.62 (d, J = 1.3 Hz, 2H),
4.57 (d, J = 11.6 Hz, 1H), 4.51 (d, J = 7.1 Hz, 2H), 4.43–4.34 (m,

6288
T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
2H), 4.31 (dd, J = 9.0, 3.5 Hz, 1H), 4.20–4.14 (m, 2H), 3.99–3.94 (m,
3H), 3.93 (d, J = 2.6 Hz, 1H), 3.76 (dd, J = 10.8, 5.7 Hz, 1H); 13C NMR
(100 MHz, CDCl3) d 147.6, 145.9, 139.8, 138.9, 138.5, 138.4, 137.6,
137.5, 133.1, 132.8, 129.3, 128.9, 128.4, 128.2, 127.6, 127.58,
127.57, 127.52, 127.3, 125.6, 121.8, 109.3, 108.1, 100.7, 88.5,
85.8, 82.4, 80.9, 76.0, 73.3, 72.5, 71.7, 71.3, 71.1, 38.8; HRMS
(ESI) calcd for C48H45ClO7 [M+Na]+791.2752, found 791.2746.
4.78–4.49 (m, 5H), 4.49–4.41 (m, 1H), 4.33–4.19 (m, 2H), 4.19–
3.84 (m, 8H), 3.81 (s, 2H), 3.73–3.62 (m, 2H), 1.45–1.30 (m, 3H);
13C NMR (400 MHz, CDCl3) d 157.1, 157.0, 154.6, 154.2, 139.0,
139.0, 138.6, 138.6, 138.1, 137.9, 137.7, 132.7, 132.3, 132.1,
131.5, 130.6, 130.3, 130.2, 130.0, 129.5, 129.4, 128.3, 128.19,
128.17, 128.15, 128.0, 127.7, 127.6, 127.54, 127.52, 127.48,
127.43, 127.29, 127.25, 125.6, 114.19, 114.17, 110.8, 110.7, 85.6,
82.4, 80.9, 79.3, 76.3, 73.3, 72.3, 71.9, 71.28, 70.5, 63.2, 55.3,
4.1.13. General procedure for the synthesis of 33j. 1,4-Anhydro-
2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-(4-ethoxybenzyl)-
37.7, 14.80; HRMS (ESI) calcd for
821.3221, found 821.3212.
C
50H51ClO7Na+ [M+Na]+
phenyl]-D-glucitol (33j)
To a solution of 27j (604 mg, 1.86 mmol) in 1:2 THF/toluene
(15 mL) under N2 gas was added n-BuLi (2.5 M in hexane, 742
4.1.16. 1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-5-(4-
isopropylbenzyl)-2-methoxyphenyl]- -glucitol (33p)
D
l
L, 1.86 mmol) dropwise at À78 °C. After 1 h, this mixture was
33p (127 mg, 0.16 mmol, 40% yield over two steps) was pre-
pared as a colorless oil from 27p (182 mg, 0.51 mmol) and 11
(220 mg, 0.41 mmol), according to the procedure for the synthesis
of 33j. Then, b form of 33p (68.9 mg, 0.086 mmol, 54% yield) was
added to a solution of 11 (1.00 g, 1.86 mmol) in toluene (10 mL)
while maintaining the temperature under À78 °C. After stirring
for 3 h under À78 °C, NH4Cl was added to quench the reaction
and the resulting mixture was extracted with CH2Cl 2. The organic
phase was removed under reduced pressure and co-evaporated
with toluene twice to afford crude 32j. The crude 32j in CH2Cl2
obtained from
a and b mixture of 33p (127 mg, 0.16 mmol) by col-
umn chromatography (6.0 g silica gel, EtOAc/toluene/n-hex-
ane = 1:1:20 to 1:1:15). 1H NMR (400 MHz, CDCl3) d 8.20–7.78
(m, 2H), 7.64–6.76 (m, 24H), 4.95–4.28 (m, 7H), 4.28–3.50 (m,
13H), 2.91–2.74 (m, 1H), 1.28–1.17 (m, 6H); 13C NMR (100 MHz,
CDCl3) d 137.1, 134.5, 133.4, 133.2, 130.0, 129.7, 128.9, 128.7,
128.3, 128.2, 128.1, 128.0, 127.6, 127.5, 127.4, 127.3, 126.2, 85.9,
80.9, 80.6, 80.6, 79.8, 77.2, 76.0, 73.4, 71.3, 71.2, 70.9, 55.4, 33.6,
29.6; HRMS (ESI) calcd for C51H53ClO6Na+ [M+Na]+ 819.3423, found
819.3389.
(5 mL) at À50 °C, was added Et3SiH (450
lL, 2.8 mmol), followed
by BF3OEt2 (220 L, 1.87 mmol) slowly to maintain the tempera-
l
ture below À40 °C. The resulting suspension was allowed to warm
to À10 °C over 1 h prior to quenching with K2CO3. After concen-
trated in vacuo, the resulting mixture was extracted with EtOAc
and washed with brine, dried over MgSO4, filtered and purified
with column chromatography (10 g silica gel, EtOAc/n-hex-
ane = 1/20) to afford
a and b mixture of 33j (264 mg, 0.34 mmol,
18% yield, two steps) as a colorless oil. 1H NMR (400 MHz, CDCl3)
d 7.51–7.28 (m, 20H), 7.26–7.19 (m, 1H), 7.18–7.05 (m, 4H), 6.84
(d, J = 8.6 Hz, 2H), 4.99–4.87 (m, 2H), 4.77–4.51 (m, 6H), 4.49–
4.28 (m, 3H), 4.27–4.17 (m, 2H), 4.16–3.95 (m, 7H), 3.81 (q,
J = 5.6 Hz, 1H), 1.43 (t, J = 5.6 Hz, 3H); 13C NMR (100 MHz, CDCl3)
d 157.2, 139.6, 138.8, 138.7, 138.5, 137.5, 137.4, 132.8, 131.1,
129.8, 129.6, 129.1, 128.8, 128.3, 128.1, 128.1,128.1, 127.8, 127.6,
127.5, 127.5, 127.4, 127.4, 127.3, 125.4, 114.2, 88.4, 85.7, 82.3,
80.8, 77.2, 75.9, 72.4, 71.6, 71.2, 70.9, 63.1, 38.2, 14.7; HRMS
(ESI) calcd for C49H49ClO6Na+ [M+Na]+ 791.3110, found 791.3110.
4.1.17. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-
5-(4-ethylbenzyl)-2-methoxyphenyl]-
D
-glucitol (33q)
a
and b mixture of 33q (94.0 mg, 0.12 mmol, 19% yield over two
steps) was prepared as
a colorless oil from 27q (273 mg,
0.80 mmol) and 11 (346 mg, 0.64 mmol), according to the proce-
dure for the synthesis of 33j. Then, b form of 33q (48 mg,
0.061 mmol, 51% yield) as a oil, was obtained from
a and b mixture
of 33q (94 mg, 0.12 mmol) by column chromatography (5.0 g silica
gel, EtOAc/toluene/n-hexane = 1/1/20 to 1/1/15). 1H NMR
(400 MHz, CDCl3) d 7.38–7.09 (m, 21H), 6.96–6.86 (m, 4H), 6.83
(s, 1H), 5.25 (s, 1H), 4.76–4.64 (m, 2H), 4.63–4.50 (m, 3H), 4.45
(d, J = 11.3 Hz, 1H), 4.35 (dd, J = 9.3, 3.3 Hz, 1H), 4.18 (s, 2H), 4.05
(d, J = 3.3 Hz, 1H), 3.98–3.88 (m, 3H), 3.85–3.79 (m, 2H), 3.76 (s,
3H), 3.58 (dd, J = 10.7, 5.2 Hz, 1H), 2.48 (q, J = 7.7 Hz, 2H), 1.11 (t,
J = 7.6 Hz, 3H); 13C NMR (100 MHz, CDCl3) d 154.7, 141.5, 139.0,
138.6, 138.2, 137.9, 137.4, 132.8, 130.4, 130.0, 129.0, 128.4,
128.3, 128.2, 128.1, 128.1, 127.6, 127.6, 127.5, 127.5, 127.4,
127.3, 127.2, 125.7, 125.2, 110.8, 82.5, 80.9,79.4, 77.8, 76.3, 73.4,
72.4, 72.1, 71.9, 71.8, 55.3, 38.2, 28.3, 15.5; HRMS (ESI) calcd for
4.1.14. 1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-3-(4-
ethoxybenzyl)-phenyl]-D-glucitol (33k)
33k (180 mg, 0.23 mmol, 16% yield, two steps) was prepared as
a colorless oil from 27k (476 mg, 1.47 mmol) and 11 (712 mg,
1.32 mmol), according to the procedure for the synthesis of
33j.1H NMR (400 MHz, CDCl3) d 7.53–7.22 (m, 21H), 7.22–7.09
(m, 6H), 4.98–4.88 (m, 2H), 4.75–4.52 (m, 5H), 4.49–4.35 (m,
2H), 4.33–4.20 (m, 2H), 4.20–4.07 (m, 3H), 4.07–3.96 (m, 2H),
3.90–3.78 (m, 1H). 2.99–2.87 (m, 1H), 1.30–1.29 (m, 6H); 13C
NMR (100 MHz, CDCl3) d 146.4, 138.9, 138.5, 138.1, 137.9, 137.7,
137.2, 136.9, 136.7, 133.9, 130.8, 128.9, 128.8, 128.4, 128.2,
128.1, 128.0, 127.6, 127.5, 127.4, 127.4, 127.3, 127.2, 126.3,
125.2, 110.4, 86.6, 82.0, 81.7, 80.3, 76.5, 73.3, 72.3, 71.9, 71.8,
C
50H51ClO6Na+ [M+Na]+ 805.3266, found 805.3234.
4.1.18. General procedure for the synthesis of compound 20a.
(1S)-1,4-Anhydro-1-[4-chloro-3-(4-methoxybenzyl)phenyl]-D-
glucitol (20a)
71.4, 38.8, 33.5, 23.9; HRMS (ESI) calcd for
C
50H51ClO5Na+
To a solution of 19a (136 mg, 0.180 mmol,) in CH2Cl2 (2 mL)
was added BF3OEt (30 L, 0.24 mmol) and EtSH (26 L,
[M+Na]+ 789.3323, found 789.3318.
l
l
0.36 mmol). After being stirred for 20 h, H2O was added to quench
reaction, and the resulting mixture was extracted with CH2Cl2 and
the aqueous layer was extracted with EtOAc. The combine organic
layer was concentrated in vacuo and the resulting residue was
purified with column chromatography (2.0 g silica gel, CH2Cl2/
MeOH = 20/1) to obtain 20a (30.6 mg, 0.077 mmol, 43% yield)) as
a colorless oil. 1H NMR (400 MHz, CD3OD) d 7.29–7.19 (m, 3H),
7.07–6.99 (m, 2H), 6.79–6.71 (m, 2H), 4.51 (d, J = 3.6 Hz, 1H),
4.11 (dd, J = 3.4, 1.6 Hz, 1H), 4.01–3.91 (m, 4H), 3.87 (dd, J = 3.6,
1.6 Hz, 1H), 3.75 (dd, J = 11.3, 2.9 Hz, 1H), 3.69 (s, 3H), 3.59 (dd,
J = 11.4, 5.3 Hz, 1H); 13C NMR (100 MHz, CD3OD) d 141.4, 140.0,
133.8, 132.9, 131.6, 131.2, 130.8, 130.2, 130.1, 126.7, 114.8, 88.1,
4.1.15. (1S)-1,4-Anhydro-2,3,5,6-tetra-O-benzyl-1-C-[4-chloro-
5-(4-ethoxybenzyl)-2-methoxyphenyl]- -glucitol (33o)
And b mixture of 33o (319 mg, 0.40 mmol, 49% yield, two
D
a
steps) was prepared as
a colorless oil from 27o (360 mg,
1.01 mmol) and 11 (436 mg, 0.81 mmol), according to the proce-
dure for the synthesis of 33j. b form of 33o (180 mg, 0.23 mmol,
58% yield) was obtained from
a and b mixture of 33o (319 mg,
0.40 mmol) using column chromatography (6.0 g silica gel,
EtOAc/n-hexane = 1:20 to 1:15). 1H NMR (400 MHz, CDCl3) d
7.52–7.02 (m, 20H), 6.97–6.83 (m, 3H), 6.81 (d, J = 6.7 Hz, 1H),
6.79–6.67 (m, 2H), 5.53–5.32 (m, 1H), 4.94–4.78 (m, 1H),

T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
6289
86.0, 82.3, 79.5, 71.4, 65.2, 55.6, 39.2; HRMS (ESI) calcd for C20H23-
ClO6 [M+Na]+ 417.1075, found 417.1075.
(dd, J = 11.3, 5.3 Hz, 1H), 2.37 (s, 3H); 13C NMR (100 MHz, CD3OD)
d 141.5, 139.5, 137.9, 137.5, 130.4, 130.3, 130.2, 128.0, 126.9, 88.1,
86.1, 82.3, 79.4, 71.4, 65.2, 39.5, 16.0; HRMS (ESI) C20 H23ClO5S
[M+Na]+ 433.0847, found 433.0869.
4.1.19. (1S)-1,4-Anhydro-1-[4-chloro-3-(3-methoxybenzyl)
phenyl]-D-glucitol (20b)
Compound 20b (8.00 mg, 0.022 mmol, 50% yield) was prepared
as a colorless oil from 19b (33 mg, 0.044 mmol), according the pro-
cedure described for the synthesis of 20a. 1H NMR (400 MHz, CD3-
OD) d 7.33–.719 (m, 3H), 7.14–7.06 (m, 1H), 6.72–6.64 (m, 3H),
4.52 (d, J = 3.7 Hz, 1H), 4.12 (dd, J = 3.4, 1.6 Hz, 1H), 4.01–3.92 (m,
4H), 3.87 (dd, J = 2.7, 1.6 Hz, 1H), 3.75 (dd, J = 11.4, 3.4 Hz, 1H),
3.67 (s, 3H), 3.59 (dd, J = 11.4, 5.4 Hz, 1H); 13C NMR (100 MHz, CD3-
OD) d 161.2, 142.4, 141.4, 139.5, 133.9, 131.6, 130.3, 130.1, 126.9,
122.2, 115.5, 112.5, 88.1, 86.1, 82.3, 79.4, 71.4, 65.2, 55.5, 40.0;
4.1.24. (1S)-1,4-Anhydro1-[4-chloro-3-(4-phenoxybenzyl)
phenyl]-D-glucitol (20g)
Compound 20g (23.4 mg, 0.051 mmol, 32% yield) was prepared
as colorless oil from 19g (131 mg, 0.16 mmol), according to the
procedure for the synthesis of 20a1H NMR (400 MHz, CD3OD) d
7.31 (d, J = 1.8 Hz, 1H), 7.28–7.21 (m, 4H), 7.13–7.08 (m, 2H),
7.02–6.97 (m, 1H), 6.91–6.85 (m,2H), 6.84–6.79 (m, 2H), 4.52 (d,
J = 3.6 Hz, 1H), 4.12 (dd, J = 3.5, 1.6 Hz, 1H), 4.02–3.92 (m, 4H),
3.87 (d, J = 3.6, 1.6 Hz, 1H), 3.76 (dd, J = 11.3, 3.0 Hz, 1H), 3.60
(dd, J = 11.4, 5.6 Hz, 1H); 13C NMR (100 MHz, CD3OD) d 158.9,
157.0, 141.5, 139.7, 136.0, 133.9, 131.2, 130.8, 130.3, 130.2,
126.9, 124.1, 119.6, 119.6, 88.1, 86.1, 82.3, 79.5, 71.4, 65.2, 39.3;
HRMS (ESI) calcd for
417.1075.
C
20H23 ClO6 [M+Na]+ 417.1075, found
4.1.20. (1S)-1,4-Anhydro-1-[4-chloro-3-(2-methoxybenzyl)
phenyl]- -glucitol (20c)
HRMS (ESI) calcd for
479.1219.
C
25H25ClO6 [M+Na]+ 479.1232, found
D
Compound 20c (32.3 mg, 0.082 mmol, 43% yield) was prepared
as a colorless oil from 19c (145 mg, 0.19 mmol), according the pro-
cedure described for the synthesis of 20a. 1H NMR (400 MHz, CD3-
OD) d 7.27–7.18 (m, 2H), 7.17–7.09 (m, 2H), 6.92–6.83 (m, 2H),
6.79–6.72 (m, 1H), 4.46 (d, J = 3.7 Hz, 1H), 4.12–4.08 (m, 1H),
3.99–3.89 (m, 4H), 3.85 (dd, J = 3.7, 1.5 Hz, 1H), 3.77–3.68 (m,
4H), 3.61–3.52 (m, 1H); 13C NMR (100 MHz, CD3OD) d 158.8,
141.0, 131.5, 131.1, 130.8, 130.1, 129.9, 128.8, 128.7, 126.5,
121.3, 111.4, 88.0, 86.0, 82.2, 79.4, 71.3, 65.1, 55.8, 34.2; HRMS
(ESI) calcd for C20H23ClO6 [M+Na]+ 417.1075, found 417.1075.
4.1.25. (1S)-1,4-Anhydro-1-[4-chloro-3-(6-methoxynaphthalen-
2-yl)methyl]phenyl]- -glucitol (20h)
D
Compound 20h (10 mg, 0.023 mmol, 15% yield) was prepared as
as colorless oil from 19h (120 mg, 0.15 mmol), according the pro-
cedure for the synthesis of 20a. 1H NMR (400 MHz, CD3OD) d
7.58 (dd, J = 13.7, 8.4 Hz, 2H), 7.45 (s, 1H), 7.36–7.18 (m, 4H),
7.11 (d, J = 2.4 Hz, 1H), 7.01 (dd, J = 9.0, 2.4 Hz, 1H), 4.51 (d,
J = 3.6 Hz, 1H), 4.18–4.07 (m, 3H), 3.97–3.97 (m, 2H), 3.88 (dd,
J = 3.6, 1.6 Hz, 1H), 3.81 (s, 3H), 3.73–3.67 (m, 1H), 3.60–3.52 (m,
1H); 13C NMR (100 MHz, CD3OD) d 158.8, 141.4, 139.7, 136.0,
134.7, 134.0, 130.5, 130.4, 130.2, 130.0, 128.8, 128.0 127.9, 126.9,
119.6, 106.6, 88.1, 86.0, 82.3, 79.4, 71.3, 65.1, 55.7, 40.0; HRMS
(ESI) calcd for C24H25ClO6 [M+Na]+ 467.1237, found 467.1244.
4.1.21. (1S)-1,4-Anhydro-1-[4-chloro-3-(4-tert-butylbenzyl)
phenyl]-D-glucitol (20d)
Compound 20d (29.0 mg, 0.069 mmol, 43% yield) was prepared
as a colorless oil from 19d (125 mg, 0.16 mmol), according to the
procedure described for the synthesis of 20a. 1H NMR (400 MHz,
CD3OD) d 7.47–7.41 (m, 3H), 7.24–7.21 (m, 2H), 7.06–7.01 (m,
2H), 4.51 (d J = 3.8 Hz, 1H), 4.12 (dd, J = 3.5, 1.7 Hz, 1H), 4.01–
3.90 (m, 4H), 3.88 (dd, J = 3.8, 1.7 Hz, 1H), 3.76 (dd, J = 11.4,
3.1 Hz, 1H), 3.60 (dd, J = 11.4, 5.4 Hz, 1H), 1.23 (s, 9H); 13C NMR
(100 MHz, CD3OD) d 141.3, 139.8, 133.9, 131.6, 130.8, 130.3,
130.1, 129.5, 126.8, 126.2, 88.1, 86.1, 82.3, 79.5, 71.4, 65.2, 44.4,
39.5, 35.1, 34.6, 31.8; HRMS (ESI) calcd for C23H30ClO5 [M+Na]+
405.0876, found 405.0876.
4.1.26. (1S)-1,4-Anhydro-1-[4-chloro-3-(3,4-methylenedioxybenzyl)
phenyl]-D-glucitol (21i)
In a solution of 19i (106 mg, 0.14 mmol) in CH2Cl2/MeOH
(20 mL, 1:4) was added Pd/C (10%, 15 mg) and H2 gas (in a balloon).
After stirred for 16 h, the mixture was filtered through a thin pad of
Celite and the filtrate was concentrated. The resulting residue was
purified with column chromatography (2.0 g silica gel, CH2Cl2/
MeOH = 20/1) to obtain 20i (20 mg, 0.049 mmol, 35% yield) as a
colorless oil.1H NMR (400 MHz, CD3OD) d 7.32–7.19 (m, 3H),
6.68–6.54 (m, 3H), 5.81 (s, 2H), 4.51 (d, J = 3.7 Hz, 1H), 4.14–4.19
(m, 1H), 4.00–3.90 (m, 4H), 3.89–3.85 (m, 1H), 3.75 (dd, J = 11.3,
2.8 Hz, 1H), 3.59 (dd, J = 11.2, 5.2 Hz, 1H); 13C NMR (100 MHz. CD3-
OD) d 149.1, 141.4, 139.8, 134.7, 133.8, 130.2, 130.1, 126.8, 122.8,
110.1, 108.9, 102.1, 88.1, 86.0, 82.3, 79.4, 71.3, 65.2, 39.6; HRMS
(ESI) calcd for C20H21ClNaO7 [M+Na]+ 431.0874, found 431.0878.
4.1.22. (1S)-1,4-Anhydro-1-[4-chloro-3-(4-fluorobenzyl)
phenyl]-D-glucitol (20e)
Compound 20e (27.6 mg, 0.072 mmol, 40% yield) was prepared
as a colorless oil from 19e (134 mg, 0.18 mmol), according the pro-
cedure for the synthesis of 20a1H NMR (400 MHz, CD3OD) d 6.34–
7.23 (m, 3H), 7.15–7.07 (m, 2H), 6.96–6.86 (m, 2H), 4.53 (d,
J = 3.7 Hz, 1H), 4.13 (dd, J = 3.5, 1.7 Hz, 1H), 4.04–3.92 (m, 4H),
3.88 (dd, J = 3.7, 1.7 Hz, 1H), 3.76 (dd, J = 11.4, 3.0 Hz, 1H), 3.61
(dd, J = 11.3, 5.4 Hz, 1H); 13C NMR (100 MHz, CD3OD) d 141.55,
139.44, 136.87, 133.86, 131.59, 131.49, 131.41, 131.21, 130.82,
130.29, 130.27, 127.04, 116.05, 115.84, 88.10, 86.08, 82.37, 79.46,
71.39, 65.22, 39.23; HRMS (ESI) calcd for C19H20ClFO5 [M+Na]+
443.1596, found 443.1598.
4.1.27. (1S)-1,4-Anhydro-1-[4-chloro-3-(4-ethoxybenzyl)
phenyl]-D-glucitol (20j)
Alpha and beta mixture of 20j (104 mg, 0.255 mmol) was pre-
pared from 33j (264 mg, 0.34 mmol) according to the procedure
for the synthesis of 20a. Then, alpha and beta mixture of 20j
(19 mg, 0.046 mmol) in pyridine (1 mL) was added Ac2O (100
lL,
1.06 mmol) and DMAP (1 mg, 0.010 mmol) and the resulting mix-
ture was stirred at ambient temperature for 3 h. After the reaction
was completed, H2O was added to quench reaction and the result-
ing mixture was extracted with EtOAc. The organic layer was then
washed with 1.0 N HCl, 1.0 N NaOH, H2O and brine and concen-
trated in vacuo. The resulting residue was purified by column chro-
matography (500 mg silica gel, EtOAc/n-hexane = 1/2 to 1/1) to
afford beta per-acetylated 20j (17 mg, 0.029 mmol, 66% yield) as
a white solid. Then, beta per-acetylated 20j in MeOH (2 mL) was
added NaOMe (3 mg) and the resulting mixture was stirred at
4.1.23. (1S)-1,4-Anhydro-1-[4-chloro-3-(4-methylthiobenzyl)
phenyl]-D-glucitol (20f)
Compound 20f (18.7 mg, 0.045 mmol, 35% yield) was prepared
as a yellowish oil from 19f (102 mg, 0.13 mmol), according to the
procedure for the synthesis of 20a. 1H NMR (400 MHz, CD3OD) d
7.30–7.21 (m, 3H), 7.15–7.08 (m, 2H), 7.07–7.01 (m, 2H), 4.52 (d,
J = 3.6 Hz, 1H), 4.12 (dd, J = 3.4, 1.6 Hz, 1H), 3.99–3.91 (m, 4H),
3.87 (dd, J = 3.6, 1.6 Hz, 1H), 3.75 (dd, J = 11.3, 2.8 Hz, 1H), 3.59

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T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
ambient temperature for 3 h. The resulting mixture was extracted
with EtOAc and the organic layer was washed with 1.0 N HCl,
NaHCO3, H2O and brine. The resulting solution was evaporated
and the residue was purified by column chromatography
(200 mg silica gel, MeOH/CH2Cl2 = 1/10) to obtain 20j (11.5 mg,
0.028 mmol, 97% yield) as a colorless oil. 1H NMR (400 MHz, CD3-
OD) d 7.29–7.18 (m, 3H), 7.04–6.98 (m, 2H), 6.77–6.71 (m, 2H),
4.51 (d, J = 3.8 Hz, 1H), 4.11 (q, J = 1.7 Hz, 1H), 3.99–3.89 (m, 6H),
3.87 (dd, J = 3.7, 1.7 Hz, 1H), 3.77–3.72 (m, 1H), 3.59 (dd, J = 11.4,
5.4 Hz, 1H), 1.29 (t, J = 7.0 Hz, 3H); 13C NMR (150 MHz, CD3OD) d
159.0, 141.5, 140.2, 134.0, 133.0, 130.9, 130.4, 130.3, 126.9,
115.6, 88.3, 86.2, 82.5, 79.6, 71.5, 65.3, 64.6, 39.3, 15.3; HRMS
(ESI) calcd for C21H25ClO6Na+ [M+Na]+ 431.1232, found 431.1231.
(320 mg, 0.59 mmol), according to the procedure for the synthesis
of 33j. Compound 20n (29.0 mg, 0.078 mmol, 28% yield, three
steps) was prepared from 33n (203 mg, 0.278 mmol), according
to the procedure for the synthesis of 20j. 1H NMR (400 MHz, CD3-
OD) d 7.20–7.13 (m, 2H), 7.05–6.98 (m, 3H), 6.97–6.92 (m, 2H),
4.48 (d, J = 4.2 Hz, 1H), 4.13 (dd, J = 3.9, 1.8 Hz, 1H), 4.00–3.95
(m,1H), 3.94–3.90 (m, 2H), 3.87 (s, 2H), 3.76 (dd, J = 11.5, 3.4 Hz,
1H), 3.60 (dd, J = 11.5, 5.9 Hz, 1H), 2.51 (q, J = 7.6 Hz, 2H), 2.11 (s,
3H), 1.12 (t, J = 7.6 Hz, 3H); 13C NMR (150 MHz, CD3OD) d 142.9,
140.2, 139.6, 139.0, 136.9, 131.1, 129.6,129.3, 128.7, 125.6, 88.6,
86.2, 82.1, 79.8, 71.5, 65.2, 40.0, 29.4, 19.4, 16.2; HRMS (ESI) calcd
for C22H O5 [M+Na]+ 395.1829, found 395.1827.
28
4.1.32. (1S)-1,4-Anhydro-1-[4-chloro-5-(4-ethoxybenzyl)-2-
4.1.28. (1S)-1,4-Anhydro-1-[4-chloro-3-(4-isopropylbenzyl)
methoxyphenyl]-D-glucitol (20o)
phenyl]-
D
-glucitol (20k)
Compound 20o (28.0 mg, 0.064 mmol, 29% yield) was prepared
as a colorless oil from 33o (170 mg, 0.22 mmol), according the pro-
cedure for the synthesis of 20a. 1H NMR (400 MHz, CD3OD) d 7.32
(d, J =13.5 Hz, 1H), 7.02-6.95 (m, 2H), 6.87 (s, 1H), 6.74–6.67 (m,
2H), 4.25 (dd, J = 13.7, 3.4 Hz, 1H), 4.10–3.84 (m, 7H), 3.77 (s,
2H), 3.75 (s, 3H), 3.58 (q, J =5.4 Hz, 1H), 1.29 (t, J = 7.0 Hz, 3H);
13C NMR (100 MHz. CD3OD) d 158.6, 133.7, 133.4, 131.6, 131.4,
130.7, 130.6, 129.5, 126.8, 115.3, 112.0, 82.4, 81.2, 79.7, 78.5,
71.5, 65.5, 64.4, 56.2, 38.5, 15.2; HRMS (ESI) C22H27ClO7Na+
[M+Na]+ 461.1338, found 461.1380.
20k (3 mg, 0.007 mmol, 15% yield over three steps) was pre-
pared as colorless oil from 33k (35.7 mg, 0.046 mmol) according
to the procedure for the synthesis of 20j. 1H NMR (400 MHz, CD3-
OD) d 7.30-7.19 (m, 2H), 7.08–7.00 (m, 5H), 4.50 (d, J = 3.7 Hz,
1H), 4.11 (q, J = 1.8 Hz, 1H), 4.04–3.89 (m, 3H), 3.86 (dd, J = 3.7,
1.5 Hz, 1H), 3.82 (s, 1H), 3.74 (dd, J = 11.3, 3.0 Hz, 1H), 3.58–3.54
(m, 1H), 2.73–2.81 (m, 1H), 1.31–1.15 (d, 6H); 13C NMR
(100 MHz, CD3OD) d 141.4, 140.3, 130.3, 130.1, 129.8, 129.8,
127.3, 127.3, 126.8, 88.1, 86.1, 82.3, 79.5, 74.3, 71.4, 42.4, 35.0,
24.4; HRMS (ESI) calcd for C22H27ClO5Na+ [M+Na]+ 429.1445, found
429.1449
4.1.33. (1S)-1,4-Anhydro-1-[4-chloro-5-(4-isopropylbenzyl)-2-
methoxyphenyl]-D-glucitol (20p)
4.1.29. (1S)-1,4-Anhydro-1-[3-(4-ethoxybenzyl)-4-methylphenyl]-
Compound 20p (17 mg, 0.039 mmol, 45% yield) was prepared as
a colorless oil from 33p (68 mg, 0.087 mmol), according to the
procedure for the synthesis of 20a. 1H NMR (600 MHz, CD3OD) d
7.06–7.00 (m, 4H), 6.91 (d, J = 1.9 Hz, 1H), 6.76 (s, 1H), 4.49 (s,
1H), 3.96–3.92 (m,1H), 3.77 (s, 3H), 3.72 (s, 3H), 3.67 (dd,
J = 11.1, 3.4 Hz, 1H), 3.64–3.58 (m, 3H), 3.57 (dd, J = 7.6, 1.7 Hz,
1H), 3.52 (dd, J = 11.3, 5.7 Hz, 1H), 2.81–2.74 (m, 1H), 1.15 (d,
J = 6.9 Hz, 6H); 13C NMR (150 MHz, CD3OD) d 157.6, 147.7, 140.7,
134.9, 133.2, 129.89, 129.87, 129.0, 128.1, 127.4, 111.6, 84.4,
82.5, 79.0, 71.5, 65.6, 64.5, 36.0, 35.1, 24.6; HRMS (ESI) calcd for
D-glucitol (20l)
Compound 33l (414 mg, 0.55 mmol, 32% yield, two steps) was
prepared as colorless oil from 27l (522 mg, 1.71 mmol) and 11
(921 mg, 1.71 mmol), according to the procedure for the synthesis
of 33j. Compound 20l (62.4 mg, 0.16 mmol, 29% yield, three steps)
was prepared from 33l (414 mg, 0.55 mmol) according to the pro-
cedure described for the synthesis of 20j. 1H NMR (400 MHz, CD3-
OD) d 7.17-7.10 (m, 2H), 7.05-6.99 (m, 1H), 6.96-6.89 (m. 2H),
6.73–6.67 (m, 2H), 4.48 (d, J = 4.2 Hz, 1H), 4.14 (dd, J = 3.8,
1.8 Hz, 1H), 4.01–3.94 (m, 1H), 3.94–3.86 (m, 4H), 3.82 (s, 2H),
3.76 (dd, J = 11.5, 3.3 Hz, 1H), 3.59 (dd, J = 11.5, 5.9 Hz, 1H), 2.09
(s, 3H), 1.27 (t, J = 7.0 Hz, 3H); 13C NMR (150 MHz, CD3OD) d
158.6, 140,6, 139.6, 137.0, 133.9,131.2, 130.7, 129.3, 125.7, 115.5,
88.7, 86.3, 82.2, 79.8, 71.6, 65.3, 64.6, 39.7, 19.5, 15.3; HRMS
(ESI) calcd for C22H28O6Na+ [M+Na]+ 411.1778, found 411.1775.
C
23H29ClO6Na+ [M+Na]+ 459.1550, found 459.1548.
4.1.34. (1S)-1,4-Anhydro-1-[4-chloro-5-(4-ethylbenzyl)-2-
methoxyphenyl]- -glucitol (20q)
D
Compound 20q (4.00 mg, 0.009 mol, 27% yield) was prepared as
a colorless oil from 33q (15.3 mg, 0.035 mmol), according to the
procedure for the synthesis of 20a. 1H NMR (400 MHz, CD3OD) d
7.37 (s, 1H), 7.02-6.97 (m, 4H), 6.87 (s, 1H), 4.90–4.88 (m, 1H),
4.06 (dd, J = 3.0, 1.1 Hz, 1H), 4.03–3.92 (m, 3H), 3.89 (s, 2H), 3.76
(s, 3H), 3.73 (dd, J = 11.7, 3.4 Hz, 1H), 3.57 (dd, J = 11.5, 5.4 Hz,
1H), 2.52 (q, J = 7.7 Hz, 2H), 1.13 (t, J = 7.7 Hz, 3H); 13C NMR
(150 MHz, CD3OD) d 156.6, 143.0, 138.6, 133.8, 131.5, 130.6,
129.7, 129.5, 128.7, 112.2, 84.2, 84.4, 82.4, 78.9, 71.3, 65.4, 39.0,
4.1.30. (1S)-1,4-Anhydro-1-[3-(4-isopropylbenzyl)-4-methylphenyl]-
D-glucitol (20m)
33m (75 mg, 0.10 mmol, 15% yield over two steps) was pre-
pared as a colorless oil from 27m (476 mg, 1.47 mmol) and 11
(712 mg, 1.32 mmol), according to the procedure for the synthesis
of 33j. Then, 20m ((10.0 mg, 0.026 mmol, 15% yield over three
steps) was prepared as
a
colorless oil from 33m (129 mg,
29.4, 15.2; HRMS (ESI) calcd for
445.1388, found 445.1368.
C
22H27ClO6Na+ [M+Na]+
0.173 mmol), according to the procedure for the synthesis of 20j.
1H NMR (400 MHz, CD3OD) d 7.22–7.03 (m, 2H), 7.03–6.90 (m,
5H), 4.78 (d, J = 4.2 Hz, 1H), 4.13 (q, J = 1.9 Hz, 1H), 4.02–3.94 (m,
1H), 3.93–3.87 (m, 2H), 3.81–3.86 (m, 2H), 3.76 (dd, J = 11.2,
3.2 Hz, 1H), 3.59 (dd, J = 11.2, 6.2 Hz, 1H), 2.79–2.69 (m, 1H), 2.09
(s, 3H), 1.13 (d, J = 6.9 Hz, 6H); 13C NMR (100 MHz, CD3OD) d
147.5, 140.2, 139.5, 139.1, 131.1, 129.5, 127.2, 125.6, 88.6, 86.2,
82.1, 79.7, 71.4, 65.2, 40.0, 34.9, 24.5, 19.4; HRMS (ESI) calcd for
4.2. In vitro human SGLT inhibition assay
Stably transfected CHO-K1 cells were used for transporter stud-
ies. SGLT was determined by uptake of [14C]-
-methyl- -glucopy-
a
D
ranoside ([14C]-AMG, specific radioactivity 310 mCi/mmol)
purchased from Perkin Elmer (Boston, USA). For the purpose of this
study, Krebs–Ringer–Henseleit (KRH) solution containing 120 mM
NaCl, 4.7 mM KCl, 1.2 mM MgCl2, 2.2 mM CaCl2, and 10 mM Hepes
(pH 7.4 with Tris) was used for SGLT uptake assays. Briefly,
hSGLT2/CHO-K1 cells or hSGLT1/CHO-K1 cells were seeded into a
white-walled 96-well culture plate (Corning, NY, USA) at a density
of 30,000 cells/well (hSGLT2) or 20,000 cells/well (hSGLT1) and
C
23H30O5Na+ [M+Na]+ 409.1991, found 411.1775.
4.1.31. (1S)-1,4-Anhydro-1-[3-(4-ethylbenzyl)-4-methylphenyl]-
D-glucitol (20n)
Compound 33n (235 mg, 0.32 mmol, 44% yield, two steps) was
prepared as a colorless oil from 27n (215 mg, 0.74 mmol) and 11

T.-S. Lin et al. / Bioorg. Med. Chem. 21 (2013) 6282–6291
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then incubated in KRH solution containing 3 l
M [14C]-AMG in
the absence or presence of inhibitors for up to 120 min at 37 °C.
At the end of the uptake period, the KRH solution was removed
and the uptake of [14C]-AMG was stopped by adding ice-cold
KRH solution (stop solution). The wells were rinsed three times
with 150 mL stop buffer. After the third rinse, the stop solution
was completely removed from the wells and the cells were solubi-
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tive[14C]-AMG using a TopCount (Perkin Elmer). The percent of
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tor concentration at half-maximal response (EC50).
Acknowledgments
22. Yao, C. H.; Song, J. S.; Chen, C. T.; Yeh, T. K.; Hung, M. S.; Chang, C. C.; Liu, Y. W.;
Yuan, M. C.; Hsieh, C. J.; Huang, C. Y.; Wang, M. H.; Chiu, C. H.; Hsieh, T. C.; Wu,
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Y.; Iida, I.; Hagima, N.; Takeuchi, H.; Chino, Y.; Asami, J.; Okumura-Kitajima, L.;
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We gratefully acknowledge the National Science Council and
Academia Sinica in Taiwan for the financial supports.We also thank
the helpful suggestions from Dr. Tsui-Ling Hsu from Academia
Sinica and Dr. Jinq-Chyi Lee from NHRI in Taiwan.
Supplementary data
Supplementary data (1D NMR spectra for compounds 20a–20q
are provided) associated with this article can be found, in the
online version, at http://dx.doi.org/10.1016/j.bmc.2013.08.067.
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