Discovery of novel acetanilide derivatives as potent and selective β3-adrenergic receptor agonists

Article abstract of DOI:10.1016/j.ejmech.2009.01.022

In the search for potent and selective human β3-adrenergic receptor (AR) agonists as potential drugs for the treatment of obesity and noninsulin-dependent (type II) diabetes, a novel series of acetanilide-based analogues were prepared and their biological activities were evaluated at the human β3-, β2-, and β1-ARs. Among these compounds, 2-pyridylacetanilide (2f), pyrimidin-2-ylacetanilide (2u), and pyrazin-2-ylacetanilide (2v) derivatives exhibited potent agonistic activity at the β3-AR with functional selectivity over the β1- and β2-ARs. In particular, compound 2u was found to be the most potent and selective β3-AR agonist with an EC₅₀ value of 0.11 μM and no agonistic activity for either the β1- or β2-AR. In addition, 2f, 2u, and 2v showed significant hypoglycemic activity in a rodent diabetic model.

Full text of DOI:10.1016/j.ejmech.2009.01.022

European Journal of Medicinal Chemistry 44 (2009) 2533–2543
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Discovery of novel acetanilide derivatives as potent and selective
receptor agonists
b3-adrenergic
*
Tatsuya Maruyama , Kenichi Onda, Masahiko Hayakawa, Tetsuo Matsui, Toshiyuki Takasu, Mitsuaki Ohta
Drug Discovery Research, Astellas Pharma Inc., 21 Miyukigaoka, Tsukuba, Ibaraki 305-8585, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
In the search for potent and selective human b3-adrenergic receptor (AR) agonists as potential drugs for
Received 22 August 2008
Accepted 22 January 2009
Available online 31 January 2009
the treatment of obesity and noninsulin-dependent (type II) diabetes, a novel series of acetanilide-based
analogues were prepared and their biological activities were evaluated at the human 3-, 2-, and 1-
ARs. Among these compounds, 2-pyridylacetanilide (2f), pyrimidin-2-ylacetanilide (2u), and pyrazin-2-
ylacetanilide (2v) derivatives exhibited potent agonistic activity at the 3-AR with functional selectivity
over the 1- and 2-ARs. In particular, compound 2u was found to be the most potent and selective 3-
AR agonist with an EC50 value of 0.11 M and no agonistic activity for either the 1- or 2-AR. In addition,
2f, 2u, and 2v showed significant hypoglycemic activity in a rodent diabetic model.
b
b
b
b
Keywords:
b
b
b
b
3-Adrenergic receptor
m
b
b
Agonist
Acetanilide
Diabetes
Ó 2009 Elsevier Masson SAS. All rights reserved.
1. Introduction
benzenesulfonanilide derivative (1) reported by Merck scientists to
be one of the first selective human b3-AR agonists [12]. Based on
In 1984,
third group of
various -agonists [1,2]. In 1989, the primary structure of the
was identified and characterized using cloning and molecular
pharmacological techniques [3–5]. 3-AR was found to play
a significant role in regulating lipolysis and thermogenesis in both
rodent and human adipose tissue. These findings suggest that 3-
AR agonists are suitable for mediating thermogenesis for the
purpose of obesity modulation and lowering the plasma glucose
and insulin levels, thereby ameliorating noninsulin-dependent
b
3-adrenergic receptors (ARs) were proposed as the
-ARs present in rat adipose tissue that utilize
3-AR
the structure of compound 1, we hypothesized that the position of
the two benzene rings in the benzenesulfonanilide moiety of
b
b
b
1 would play an important role in its
b3-AR agonistic activity and/or
its selectivity over 1- and 2-ARs. We also assumed that the
b
b
b
relative position of the two benzene rings on both benzenesulfo-
nanilide and phenylacetanilide would be similar to that shown in
Fig. 2. Thus, we planned to design and synthesize the phenyl-
acetanilide derivative (2a) by converting the sulfonamide moiety of
1 into an acetamide moiety. Since compound 2a showed good
b
b3-AR agonistic activity, further attempts at modification were
(type II) diabetes. Subtype selectivity for
must be kept in mind since activation of the
b
3-AR agonists specifically
1- or 2-ARs would
made (Fig. 3). In this paper, we describe the synthesis and struc-
ture–activity relationships (SARs) of these newly designed acet-
b
b
cause undesirable side effects such as increased heart rate or
muscle tremors. A number of laboratories have been engaged in
anilide-based b3-AR agonists.
developing potent and selective
obesity and type II diabetes [6,7]. Early potent and selective rat
b
3-AR agonists for the treatment of
2. Chemistry
b3-
AR agonists, such as BRL-37344 [8] and CL-316243 [9], were
reported to be effective anti-obesity and anti-diabetic agents in
rodents (Fig. 1) [10]. However, human clinical trials with these
drugs for the treatment of metabolic disorders have been disap-
pointing due to a lack of selectivity and/or potency or poor
pharmacokinetics [11].
Acetanilides 2a–h were prepared from aniline intermediate 5, as
illustrated in Scheme 1. Compound 5 was synthesized from (S)-2-
[(4-benzyloxyphenoxy)methyl]oxirane (3) [13] and N-benzyl-N-[2-
(4-nitrophenyl)ethyl]amine [14], followed by reduction of the nitro
group with iron. Compound 5 was coupled with the appropriate
acetyl chlorides or acetic acids, followed by deprotection of the
benzyl group to afford the desired products 2a–h. Another route
starting from aniline intermediate 11 is illustrated in Scheme 2.
(S)-2-{[4-(Methoxymethoxy)phenoxy]methyl}-oxirane (8a) was
prepared from 4-(methoxymethoxy)phenol (7a) [15] and (R)-
epichlorohydrin. Compound 8a was treated with a commercially
As our lead compound in the search for novel potent and
selective
human
b
3-AR
agonists,
we
chose
the
* Corresponding author. Tel.: þ81 29 863 6749; fax: þ81 29 852 5387.
E-mail address: tatsuya.maruyama@jp.astellas.com (T. Maruyama).
0223-5234/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2009.01.022

2534
T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
OH
partial
tion of a substituent at the
2a was tolerated while maintaining
this change may not improve functional selectivity over
b
1-AR agonism. These results indicated that the introduc-
-position of the phenylacetyl moiety in
3-AR agonistic activity, but
1-AR.
H
N
OH
H
N
O
Cl
Cl
O
O
a
ONa
ONa
OH
O
b
O
O
b
We then examined the effects of the substituents on the phenyl
ring of the phenylacetyl moiety in 2a. The introduction of the chloro
group as an electron-withdrawing group on the phenyl ring of the
phenylacetyl moiety (2k–m) resulted in slightly increased agonistic
BRL-37344
CL-316243
Fig. 1. Chemical structure of early b3-AR agonists.
activity at the
seemed to be ortho-chloro (EC50 ¼ 0.74
(EC50 ¼ 0.89 M) > para-chloro (EC50 ¼ 1.2 M); however
b
3-AR relative to 2a, and the rank of order of potency
M) > meta-chloro
1-AR
available 2-(4-nitrophenyl)ethylamine hydrochloride, followed by
protection of the amine with a tert-butoxycarbonyl (Boc) group to
provide nitro compound 10. Hydrogenation of 10 yielded aniline
intermediate 11. The coupling of 11 with the appropriate acetyl
chlorides or acetic acids, followed by deprotection of the methox-
ymethyl (MOM) and Boc groups afforded the desired products 2i–r.
An alternate route was developed for the synthesis of acetanilides
2s–v, as illustrated in Scheme 3, because the corresponding acetic
acids were unstable. 4-Aminobenzylcyanide (13) was treated with
the appropriate esters in refluxing xylene to provide anilides 14a–d.
Anilides 14a–d was then hydrogenated in the presence of Raney-
nickel, followed by coupling with (S)-2-{[4-(2-methoxyethox-
ymethoxy)phenoxy]methyl}-oxirane (8b) to afford compounds
15a–d. Deprotection of the 2-methoxyethoxymethyl (MEM) group
in 15a–d yielded the desired products 2s–v.
m
m
m
b
agonistic activity did not decrease for any of these. The methoxy
group as an electron-donating group was also introduced on the
phenyl ring of the phenylacetyl moiety (2n–p), which yielded
results similar to those of the chlorophenyl derivatives, the intro-
duction of an ortho-methoxy group (2n) led to a 5-fold increase in
agonistic activity at the
to 2a. These results revealed that the introduction of a substituent
on the phenyl ring of the phenylacetyl moiety in 2a allowed the
b
3-AR (EC50 ¼ 0.36 M, IA ¼ 0.88) relative
m
potency of
b3-AR to be maintained, but may not be efficacious for
improving functional selectivity over
b
1-AR.
Next, the modification of the phenyl ring in the phenylacetyl
moiety of 2a into other aromatic rings was investigated as shown in
Table 2. Replacement of the phenyl ring with 1-naphthyl (2c) and
indol-3-yl (2e) ring resulted in slightly increased agonistic activity
3. Results and discussion
(3-fold) at the
b3-AR relative to 2a without decreasing b1-AR
agonistic activity. The 2-naphthylacetanilide derivative (2d)
showed an improvement in potency of more than 5-fold for both
All compounds listed in Tables 1 and 2 were evaluated for their
agonistic activities in stimulating an increase in cyclic AMP (cAMP)
levels in Chinese hamster ovary (CHO) cells expressing the cloned
b3- and b1-AR relative to 2a. The thenylacetanilide derivatives
(2q,r) showed results similar to those of 2e. These results suggested
human
b
3-,
b
2-, and
b
1-ARs. The results for reference compounds,
-AR agonist) and compound 1,
that altering the aromatic rings may not be sufficient to decrease
agonistic activity at the b1-AR.
isoproterenol (ISO; non-selective
b
are also shown for comparison in Table 1.
Replacement of the phenyl ring with a pyridine ring yielded
interesting results. The agonistic activity of the 2-pyridylacetanilide
The phenylacetanilide derivative (2a) of our lead compound was
found to be a modestly potent
IA ¼ 0.91) with a lower potency for the
however compound 2a also showed modest
activity (EC50 ¼ 0.76 M, IA ¼ 0.50). In order to improve
agonistic activity and functional selectivity over 1-AR, modifica-
tion of the phenylacetyl moiety in 2a was examined.
Initially, introduction of substituent at the -position of the
phenylacetyl moiety in 2a was investigated as shown in Table 1.
Introduction of the phenyl group at the -position of the phenyl-
acetyl moiety (2b) resulted in a considerable increase in agonistic
activity (14-fold) at the M, IA ¼ 1.05) and a
3-AR (EC50 ¼ 0.12
4-fold increase in agonistic activity at the M,
1-AR (EC50 ¼ 0.18
IA ¼ 0.65) relative to that of 2a. This indicated that 2b was a non-
selective 3- and 1-AR agonist. Next, the introduction of
a hydrogen bond donor or acceptor at the -position of the phe-
b
3-AR agonist (EC50 ¼ 1.7
2-AR (EC50 ¼ >100
1-AR agonistic
3-AR
m
M,
derivative (2f) increased 6-fold at the
b
b
3-AR (EC50 ¼ 0.29
1-AR. In addition its
M, IA ¼ 0.14) relative to 2a,
3-
mM,
b
mM);
IA ¼ 0.74), but decreased 3.5-fold at the
b
intrinsic activity decreased (EC50 ¼ 2.7
m
m
b
which indicated that 2f was a potent and functionally selective
b
b
AR agonist. In contrast, the intrinsic activity of the 3-pyr-
idylacetanilide (2g) and 4-pyridylacetanilide (2h) derivatives
a
decreased at the
2a without decreasing agonistic activity at the
suggested that changing the position of the nitrogen atom on the 2-
pyridyl moiety in 2f would result in both potency at the 3-AR and
functional selectivity over 1-AR; this promoted us to synthesize
b3-AR (IA ¼ 0.48 and 0.49, respectively) relative to
b
1-AR. These results
a
b
b
m
b
b
m
benzene-fused 2-pyridyl analogues. The results obtained for qui-
nolin-2-ylacetanilide derivative (2s) were similar to those of 2f,
with EC50 values of 0.65 mM at the b3-AR and 1.7 mM at the b1-AR,
respectively; however, 2s was less potent and less functionally
selective than 2f. The agonistic activity of the isoquinolin-3-ylace-
b
b
a
nylacetyl moiety in 2a was examined, because the sulfonanilide
moiety of 1 possessed two oxygen atoms. Both the hydroxyl (2i)
and amino (2j) derivatives exhibited a slight increase in agonistic
tanilide derivative (2t) at the
much greater than that of 2a, but it did not decrease
b
3-AR (EC50 ¼ 0.14
m
M, IA ¼ 0.77) was
b
1-AR
activity (1.8-fold) at the
activity relative to 2a. The introduction of a hydroxyl group to 2a
led to full agonistic activity (IA ¼ 1.12) at the
b3-AR without diminishing b1-AR agonistic
agonistic activity (EC50 ¼ 0.35
m
M, IA ¼ 0.36).
Lastly, we replaced the phenyl ring with a heteroaromatic ring
containing two nitrogens. The pyrimidin-2-ylacetanilide derivative
b1-AR, with only
(2u) increased agonistic activity at the
(EC50 ¼ 0.11 M, IA ¼ 0.57), relative to 2a, and dramatically
decreased agonistic activity at the M), which
b3-AR 15-fold
m
O O
b1-AR (EC50 ¼ >100
m
S
N
suggested that the two nitrogen atoms at 1- and 3-positions on the
pyrimidin-2-yl moiety may play a very important role in the
H
improvement of
1-AR agonistic activity. Furthermore, the pyrazin-2-ylacetanilide
derivative (2v) exhibited a 20-fold increase in agonistic activity at
the M, IA ¼ 0.64) relative to 2a, and was the
3-AR (EC50 ¼ 0.083
most potent 3-AR agonist in this study. Compound 2v induced
b3-AR agonistic activity and the substantial loss of
O
b
N
H
b
m
Fig. 2. Flexible alignment of phenylacetanilide and benzenesulfonanilide.
b

T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
2535
OH
OH
H
N
H
N
O
O
O O
S
H
O
Ar
N
HO
N
H
HO
R
2
1
Fig. 3. Design of acetanilides based on compound 1.
a 4-fold decrease in agonistic activity at the
b
1-AR (EC50 ¼ 3.1
m
M,
5.1.1. (S)-1-{N-Benzyl-N-[2-(4-nitrophenyl)ethyl]amino}-3-(4-
benzyloxyphenoxy)-2-propanol (4)
IA ¼ 0.18) relative to 2a and its potency for
b3-AR and functional
selectivity over
b
1-AR was comparable to that of compound 1.
A mixture of (S)-2-[(4-benzyloxyphenoxy)methyl]oxirane (3)
(3.48 g) and N-benzyl-N-[2-(4-nitrophenyl)ethyl]amine (3.22 g) in
2-propanol (70 mL) was refluxed for 4 h. After cooling to room
temperature, the resultant mixture was concentrated in vacuo. The
residue was purified using column chromatography on silica gel
with n-hexane/EtOAc (3:1) as the eluent to yield 4 (5.91 g) as
Given the results of the in vitro study, compounds 2f, 2u, and 2v
were selected for in vivo evaluation in a rodent model of type II
diabetes (Table 3). The compounds were administered orally for 4
days in diabetic kk mice and the effects on plasma glucose were
measured. All compounds exhibited a significant reduction in
plasma glucose levels at the dose of 30 mg/kg. In particular, 2f
showed the most potent hypoglycemic activity (32% decrease).
a colorless oil. 84% yield; 1H NMR (CDCl3)
d: 2.69–2.93 (6H, m), 3.05
(3H, br s), 3.60 (1H, d, J ¼ 13.4 Hz), 3.82 (1H, d, J ¼ 13.4 Hz), 3.85–
3.88 (2H, m), 3.97–4.01 (1H, m), 5.02 (2H, s), 6.79 (2H, d, J ¼ 9.2 Hz),
6.89 (2H, d, J ¼ 9.2 Hz), 7.20–7.43 (12H, m), 8.08 (2H, d, J ¼ 8.6 Hz);
MS (FAB) m/z: 513 (MHþ).
4. Conclusion
We have identified a new series of acetanilide-based
b3-AR
5.1.2. (S)-1-{N-[2-(4-Aminophenyl)ethyl]-N-benzylamino}-3-(4-
benzyloxyphenoxy)-2-propanol (5)
agonists and described their synthesis and SARs. Among these
compounds, the 2-pyridylacetanilide (2f), pyrimidin-2-ylacetani-
lide (2u), and pyrazin-2-ylacetanilide (2v) derivatives showed
To a solution of 4 (4.07 g) in methanol (60 mL) were added 2 M
HCl aqueous solution (8 mL) and iron powder (0.45 g), and the
mixture was refluxed for 1 h. After cooling to room temperature,
the excess iron was removed by filtration, and the filtrate was
concentrated in vacuo. The residue was partitioned between ethyl
acetate and 1 M NaOH aqueous solution, and the organic layer was
washed with water and brine, and then dried and concentrated in
vacuo to yield 5 (3.62 g) as a colorless oil. 94% yield; 1H NMR (CDCl3)
potent agonistic activity at the
0.083 M, respectively), with functional selectivity over
ARs. With agonistic activity almost completely abolished at both
the 1- and 2-ARs, the pyrimidin-2-ylacetanilide derivative (2u)
was found to be the most potent and selective 3-AR agonist in this
b
3-AR (EC50 ¼ 0.29, 0.11, and
m
b
1- and 2-
b
b
b
b
study. In addition, these compounds exhibited significant hypo-
glycemic activity in diabetic kk mice.
d: 2.63–2.82 (6H, m), 3.25 (1H, br s), 3.53 (2H, br s), 3.59 (1H, d,
J ¼ 13.4 Hz), 3.83–3.87 (3H, m), 3.94–3.97 (1H, m), 5.01 (2H, s), 6.59
(2H, d, J ¼ 7.3 Hz), 6.80 (2H, d, J ¼ 7.9 Hz), 6.87–6.94 (4H, m), 7.24–
7.42 (10H, m); MS (FAB) m/z: 483 (MHþ).
5. Experimental
5.1. Chemistry
5.1.3. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2-phenylacetanilide (6a)
Melting points were determined with a Yanaco MP-500D
melting point apparatus and are uncorrected. 1H NMR spectra were
recorded on a JEOL EX90, EX400, or GX500 spectrometer, and the
To a solution of 5 (0.38 g) and triethylamine (0.86 g) in chloro-
form (7 mL) was added phenylacetyl chloride (0.13 g) at 0 ꢁC, and
the mixture was stirred at room temperature for 1 h. The resultant
mixture was partitioned between chloroform and water. The
organic layer was washed with water and brine, and then dried and
concentrated in vacuo. The residue was purified using column
chromatography on silica gel with chloroform as the eluent to yield
chemical shifts are expressed in
d (ppm) values with tetrame-
thylsilane as an internal standard (NMR description key: s ¼ singlet,
d ¼ doublet, t ¼ triplet, m ¼ multiplet, and br ¼ broad peak). Mass
spectra were recorded on a Hitachi M-80 or JEOL JMS-DX300
spectrometer. The elemental analyses were performed with
a Yanaco MT-5 microanalyzer (C, H, N) and were within ꢀ0.4% of
the theoretical values. During the work-up, all organic solutions
were dried over anhydrous Mg2SO4.
4 (0.43 g) as a colorless powder. 91% yield; 1H NMR (CDCl3)
d: 2.62–
2.82 (6H, m), 3.16 (1H, br s), 3.58 (1H, d, J ¼ 13.4 Hz), 3.72 (2H, s),
OH Bn
OH Bn
O
O
N
O
N
b
O
a
NO2
NH2
BnO
BnO
BnO
5
3
4
Bn
N
OH
6a : Ar = Ph, R = H
6b : Ar = Ph, R = Ph
6e : Ar = indol-3-yl, R = H
6f : Ar = 2-pyridyl, R = H
6g : Ar = 3-pyridyl, R = H
6h : Ar = 4-pyridyl, R = H
O
c
d
O
Ar
N
H
BnO
HO
6c : Ar = 1-naphthyl, R =H
6d : Ar = 2-naphthyl, R = H
R
6a-h
OH
H
N
O
2a : Ar = Ph, R =H
2e : Ar = indol-3-yl, R =H
2f : Ar = 2-pyridyl, R = H
2g : Ar = 3-pyridyl, R = H
2h : Ar = 4-pyridyl, R = H
O
Ar
2b : Ar = Ph, R = Ph
N
2c : Ar = 1-naphthyl, R = H
2d : Ar = 2-naphthyl, R = H
H
R
2a-h
Scheme 1. Reagents and conditions: (a) N-benzyl-N-[2-(4-nitrophenyl)ethyl]amine, iPrOH, reflux; (b) Fe, 2 M HCl aq., MeOH; (c) ArCH(R)COCl, Et3N, CHCl3; or ArCH2CO2H, EDC$HCl,
HOBt, DMF; (d) H2, Pd/C, EtOH, and then 4 M HCl–EtOAc, MeOH.

2536
T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
OH
H
N
O
O
OH
7a,b
O
b
c
a
NO2
MOMO
RO
RO
9
8a,b
8a : R = MOM
8b : R = MEM
OH Boc
N
OH Boc
O
N
O
d
NO2
NH2
MOMO
MOMO
10
11
OH
OH
Boc
N
H
N
O
O
e
O
O
f
Ar
Ar
N
H
N
H
HO
MOMO
R
12a-j
R
2i-r
2i : Ar = Ph, R = OH
2j : Ar = Ph, R = NH2
12a : Ar = Ph, R = OH
12b : Ar = Ph, R = NHBoc
12c : Ar = 2-ClC6H4, R = H
12d : Ar = 3-ClC6H4, R = H
12e : Ar = 4-ClC6H4, R = H
2k : Ar = 2-ClC6H4, R = H
2l : Ar = 3-ClC6H4, R = H
2m : Ar = 4-ClC6H4, R = H
12f : Ar = 2-MeOC6H4, R = H
12g : Ar = 3-MeOC6H4, R = H
12h : Ar = 4-MeOC6H4, R = H
2n : Ar = 2-MeOC6H4, R = H
2o : Ar = 3-MeOC6H4, R = H
2p : Ar = 4-MeOC6H4, R = H
12i : Ar = 2-thenyl, R = H
12j : Ar = 3-thenyl, R = H
2q : Ar = 2-thenyl, R = H
2r : Ar = 3-thenyl, R = H
Scheme 2. Reagents and conditions: (a) (R)-epichlorohydrin, KOH aq.; (b) 2-(4-nitrophenyl)ethylamine hydrochloride, Et3N, iPrOH, reflux; (c) (Boc)2O, THF; (d) H2, Pd/C, EtOH; (e)
ArCH(R)CO2H, EDC$HCl, HOBt, DMF; or ArCH2COCl, Et3N, CHCl3; (f) 4 M HCl–EtOAc, MeOH.
3.78–3.85 (3H, m), 3.93–3.96 (1H, m), 5.01 (2H, s), 6.78 (2H, d,
J ¼ 9.1 Hz), 6.86–6.91 (2H, m), 6.99–7.03 (2H, m), 7.21–7.42 (17H,
m); MS (FAB) m/z: 601 (MHþ).
d
: 2.62–2.78 (6H, m), 3.55 (1H, d, J ¼ 13.5 Hz), 3.78–3.84 (2H, m),
3.91–3.94 (1H, m), 4.17 (2H, s), 5.00 (2H, s), 6.77 (2H, d, J ¼ 9.1 Hz),
6.86–6.90 (2H, m), 6.96 (2H, d, J ¼ 8.6 Hz), 7.15–7.56 (17H, m), 7.86–
7.91 (2H, m), 8.02 (1H, d, J ¼ 8.0 Hz); MS (FAB) m/z: 651 (MHþ).
5.1.4. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2,2-diphenylacetanilide (6b)
The title compound was prepared in the same manner as
described for 6a using diphenylacetyl chloride instead of phenyl-
5.1.6. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2-(2-naphthyl)acetanilide (6d)
To a solution of 5 (0.8 g) and 2-naphthylacetic acid (0.31 g) in
N,N-dimethylformamide (10 mL) were added 1-ethyl-3-(30-dime-
acetyl chloride as a colorless powder. 86% yield; 1H NMR (CDCl3)
d:
2.63–2.83 (6H, m), 3.17 (1H, br s), 3.58 (1H, d, J ¼ 13.4 Hz), 3.80–3.86
(3H, m), 3.94–3.97 (1H, m), 5.00 (2H, s), 5.06 (1H, s), 6.78 (2H, d,
J ¼ 9.2 Hz), 6.88 (2H, d, J ¼ 9.2 Hz), 7.03 (2H, d, J ¼ 8.5 Hz), 7.21–7.42
(22H, m); MS (FAB) m/z: 677 (MHþ).
thylaminopropyl)carbodiimide
hydrochloride
(0.32 g)
and
1-hydroxybenzotriazole (0.22 g), and the mixture was stirred at
room temperature for 13 h. The resulting mixture was concentrated
in vacuo, and partitioned between ethyl acetate and NaHCO3
aqueous solution. The organic layer was dried and concentrated in
vacuo. The residue was purified using column chromatography on
silica gel with n-hexane/EtOAc (3:2) as the eluent to yield 6d
5.1.5. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2-(1-naphthyl)acetanilide (6c)
The title compound was prepared in the same manner as
described for 6a using 1-naphthylacetyl chloride instead of phe-
nylacetyl chloride as a colorless powder. 51% yield; 1H NMR (CDCl3)
(0.61 g) as a colorless powder. 57% yield; 1H NMR (CDCl3)
d: 2.61–
2.83 (6H, m), 3.56 (1H, br s), 3.57 (1H, d, J ¼ 13.4 Hz), 3.78–3.95 (6H,
m), 5.00 (2H, s), 6.77 (2H, d, J ¼ 9.1 Hz), 6.87 (2H, d, J ¼ 9.1 Hz), 7.00
OH
H
N
O
NC
O
b,c
NC
a
O
Ar
Ar
N
H
NH2
N
H
MEMO
15a-d
13
14a-d
15a : Ar = quinolin-2-yl
14a : Ar = quinolin-2-yl
15b : Ar = isoquinolin-3-yl
15c : Ar = pyrimidin-2-yl
15d : Ar = pyrazin-2-yl
14b : Ar = isoquinolin-3-yl
14c : Ar = pyrimidin-2-yl
14d : Ar = pyrazin-2-yl
OH
H
N
O
2s: Ar = quinolin-2-yl
2t : Ar = isoquinolin-3-yl
2u : Ar = pyrimidin-2-yl
2v : Ar = pyrazin-2-yl
d
O
Ar
N
H
HO
2s-v
Scheme 3. Reagents and conditions: (a) ArCH2CO2Et, xylene, reflux; (b) H2, Raney-Ni, conc. NH3 aq., EtOH–THF; (c) 8b, 2-propanol, reflux; (d) 4 M HCl–EtOAc, MeOH.

T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
2537
Table 1
Table 2 (continued )
b
-AR agonistic activity of substituted phenylacetanilide derivatives.
Compound
Ar
EC50
,
m
Ma (IAb)
OH
H
N
O
b
3-AR
b2-AR
b1-AR
O
R1
N
H
HO
R2
N
N
2g
2h
1.5 (0.48)
>100 (0)
0.91 (0.35)
Compound
R1
R2
EC50, m
Ma (IAb)
b
3-AR
1.7 (0.91)
b
2-AR
b
1-AR
2a
2b
2i
2j
2k
2l
2m
2n
2o
2p
H
H
H
H
H
>100 (0.03)
>100 (0.02)
>100 (0)
0.76 (0.50)
0.18 (0.65)
0.59 (1.12)
0.66 (0.50)
0.72 (0.51)
0.52 (0.49)
0.65 (0.71)
>100 (0.03)
0.26 (0.38)
1.7 (0.29)
Ph
OH
NH2
H
H
H
H
H
H
0.12 (1.05)
0.96 (0.82)
0.96 (0.68)
0.74 (0.90)
0.89 (0.96)
1.2 (1.10)
0.36 (0.88)
0.56 (0.85)
1.7 (0.86)
>100 (0)
2-Cl
3-Cl
4-Cl
2-MeO
3-MeO
4-MeO
>100 (0.01)
>100 (0.03)
>100 (0.02)
>100 (0)
>100 (0.01)
>100 (0.01)
0.083 (0.55)
0.19 (0.47)
0.96 (0.69)
0.71 (0.84)
0.28 (0.49)
2s
>100 (0.01)
>100 (0.02)
N
ISO
1
0.10 (1.00)
0.078 (0.73)
0.003 (1.00)
>100 (0.02)
0.012 (1.00)
1.6 (0.21)
2t
0.14 (0.77)
0.11 (0.57)
0.35 (0.36)
N
a
Agonistic activity was assessed by measuring cAMP accumulation in CHO cells
-ARs.
Values in parentheses represent the intrinsic activity (IA) given as a fraction of
the maximum stimulation with isoproterenol.
expressing
b
b
N
2u
N
>100 (0)
>100 (0.07)
N
N
2v
0.083 (0.64)
>100 (0.01)
3.1 (0.18)
Table 2
a
b
-AR agonistic activity of arylacetanilide derivatives.
Agonistic activity was assessed by measuring cAMP accumulation in CHO cells
-ARs.
Values in parentheses represent the intrinsic activity (IA) given as a fraction of
the maximum stimulation with isoproterenol.
expressing
b
OH
H
N
b
O
O
Ar
N
H
HO
(2H, d, J ¼ 7.9 Hz), 7.08 (1H, s), 7.21–7.52 (14H, m), 7.52–7.87 (3H,
m); MS (FAB) m/z: 651 (MHþ).
Compound
Ar
EC50
,
m
Ma (IAb)
b3-AR
b2-AR
b
1-AR
5.1.7. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2-(3-indolyl)acetanilide (6e)
The title compound was prepared in the same manner as
described for 6d using 3-indolylacetic acid instead of 2-naphthyl-
2a
1.7 (0.91)
>100 (0.03)
>100 (0)
0.76 (0.50)
acetic acid as a colorless powder. 52% yield; 1H NMR (CDCl3)
d:
2.61–2.78 (6H, m), 3.56–3.63 (2H, m), 3.81–3.89 (4H, m), 5.01 (2H,
s), 5.20–5.22 (1H, m), 6.67 (2H, d, J ¼ 8.5 Hz), 6.83–6.86 (2H, m),
6.92 (2H, d, J ¼ 8.5 Hz), 7.03–7.07 (2H, m), 7.14–7.44 (13H, m), 7.55–
7.62 (2H, m), 8.09 (1H, s), 8.21 (1H, s); MS (FAB) m/z: 640 (MHþ).
2c
0.47 (0.96)
0.60 (0.47)
5.1.8. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2-(2-pyridyl)acetanilide (6f)
The title compound was prepared in the same manner as
described for 6d using 2-pyridylacetic acid instead of 2-naphthyl-
2d
2e
0.24 (0.90)
0.49 (1.01)
>100 (0.01)
>100 (0)
0.15 (0.40)
0.55 (0.28)
H
N
acetic acid as a colorless powder. 52% yield; 1H NMR (CDCl3)
d: 2.62–
2.87 (6H, m), 3.22 (1H, br s), 3.59 (1H, d, J ¼ 13.2 Hz), 3.80–3.88 (5H,
m), 3.92–4.00 (1H, m), 5.00 (2H, s), 6.75–6.82 (2H, m), 6.85–6.90 (2H,
m), 7.02–7.07 (2H, m), 7.20–7.47 (8H, m), 7.65–7.72 (1H, m), 8.58–
8.65 (1H, m), 9.71 (1H, br s); MS (FAB) m/z: 602 (MHþ).
S
2q
2r
0.50 (0.79)
0.55 (0.80)
>100 (0)
0.55 (0.28)
0.49 (0.37)
S
Table 3
Oral hypoglycemic activity in kk mice.
>100 (0.04)
Compound
Percent reduction in plasma glucosea
2f
2u
2v
32**,b
20*
2f
0.29 (0.74)
>100 (0)
2.7 (0.14)
13**
N
a
The compounds were administered orally to male kk mice for 4 days at the dose
of 30 mg/kg.
b
(continued on next page)
Statistically significant at *p < 0.05, **p < 0.01.

2538
T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
5.1.9. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2-(3-pyridyl)acetanilide (6g)
The title compound was prepared in the same manner as
described for 6d using 3-pyridylacetic acid instead of 2-naphthyl-
C29H30N2O4$HCl: C, 68.70; H, 6.16; N, 5.53; Cl, 6.99. Found: C, 68.41;
H, 6.09; N, 5.48; Cl, 7.09.
5.1.14. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(2-naphthyl)acetanilide
hydrochloride (2d)
acetic acid as a colorless powder. 61% yield; 1H NMR (CDCl3)
d:
2.60–2.85 (6H, m), 3.18 (1H, br s), 3.59 (1H, d, J ¼ 13.5 Hz), 3.68 (2H,
s), 3.80–3.85 (3H, m), 3.93–3.96 (1H, m), 5.01 (2H, s), 6.78 (2H, d,
J ¼ 9.0 Hz), 6.88 (2H, d, J ¼ 9.5 Hz), 7.05 (2H, d, J ¼ 9.0 Hz), 7.19–7.42
(13H, m), 7.72 (1H, d, J ¼ 8.0 Hz), 8.55–8.57 (2H, m); MS (FAB) m/z:
602 (MHþ).
The title compound was prepared in the same manner as
described for 2a using 6d instead of 6a as a colorless solid. 35%
yield; mp 258–260 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.93–
3.03 (3H, m), 3.12–3.19 (3H, m), 3.81–3.90 (4H, m), 4.17 (1H, br s),
5.84 (1H, d, J ¼ 4.9 Hz), 6.69 (2H, d, J ¼ 8.8 Hz), 6.77 (2H, d,
J ¼ 9.3 Hz), 7.18 (2H, d, J ¼ 8.3 Hz), 7.46–7.53 (3H, m), 7.60 (2H, d,
J ¼ 8.8 Hz), 7.84–7.90 (4H, m), 9.00 (3H, br s),10.39 (1H, s); MS (FAB)
m/z: 471 (MHþ); Anal. Calcd for C29H30N2O4$HCl: C, 68.70; H, 6.16;
N, 5.53; Cl, 6.99. Found: C, 68.76; H, 6.24; N, 5.52; Cl, 6.92.
5.1.10. (S)-40-(2-{N-Benzyl-N-[3-(4-benzyloxyphenoxy)-2-
hydroxypropyl]amino}ethyl)-2-(4-pyridyl)acetanilide (6h)
The title compound was prepared in the same manner as
described for 6d using 4-pyridylacetic acid instead of 2-naphthyl-
acetic acid as a colorless powder. 69% yield; 1H NMR (CDCl3)
d:
2.60–2.89 (6H, m), 3.17 (1H, br s), 3.59 (1H, d, J ¼ 13.6 Hz), 3.66 (2H,
s), 3.77–3.87 (3H, m), 3.90–3.98 (1H, m), 5.00 (2H, s), 6.74–6.79 (2H,
m), 6.85–6.89 (2H, m), 7.03–7.08 (2H, m), 7.22–7.43 (14H, m), 8.58–
8.60 (2H, m); MS (FAB) m/z: 602 (MHþ).
5.1.15. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(indol-3-yl)acetanilide
hydrochloride (2e)
The title compound was prepared in the same manner as
described for 2a using 6e instead of 6a as a colorless solid. 66%
5.1.11. (S)-40-(2-{[2-Hydroxy-3-(4-
yield; mp 204–206 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.91–
hydroxyphenoxy)propyl]amino}ethyl)-2-phenylacetanilide
hydrochloride (2a)
3.05 (3H, m), 3.10–3.20 (3H, m), 3.73 (2H, s), 3.81–3.90 (2H, m), 4.16
(1H, br s), 5.83 (1H, d, J ¼ 4.9 Hz), 6.69 (2H, d, J ¼ 9.3 Hz), 6.77 (2H, d,
J ¼ 9.3 Hz), 6.97 (1H, t, J ¼ 7.3 Hz), 7.06 (1H, t, J ¼ 7.3 Hz), 7.17 (2H, d,
J ¼ 8.3 Hz), 7.25 (1H, d, J ¼ 2.0 Hz), 7.35 (2H, d, J ¼ 8.3 Hz), 7.57–7.62
(3H, m), 8.78–8.99 (3H, m), 10.18 (1H, s), 10.93 (1H, s); MS (FAB) m/
z: 460 (MHþ); Anal. Calcd for C27H29N3O4$HCl: C, 65.38; H, 6.10; N,
8.47; Cl, 7.15. Found: C, 65.20; H, 6.05; N, 8.43; Cl, 6.93.
To a mixture of 6a (0.41 g) in ethanol (50 mL) was added
palladium on carbon (10% w/w, 0.1 g), and the mixture was stirred
under hydrogen atmosphere for 15 h. The catalyst was removed
by filtration, and the filtrate was concentrated in vacuo. The
residue was dissolved in methanol (10 mL), and 4 M HCl–EtOAc
solution (0.2 mL) was added, and then the mixture was concen-
trated in vacuo. The crude solid was purified by recrystallization
from ethanol to yield 2a (0.1 g) as a colorless solid. 32% yield; mp
5.1.16. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(2-pyridyl)acetanilide
hydrochloride (2f)
243–245 ꢁC (EtOH); 1H NMR (DMSO-d6)
d: 2.88–3.03 (3H, m),
3.10–3.20 (3H, m), 3.63 (2H, s), 3.80–3.90 (2H, m), 4.05–4.20 (1H,
m), 5.82 (1H, d, J ¼ 4.9 Hz), 6.68 (2H, d, J ¼ 8.4 Hz), 6.77 (2H, d,
J ¼ 8.8 Hz), 7.18 (2H, d, J ¼ 8.3 Hz), 7.22–7.26 (1H, m), 7.29–7.33
(4H, m), 7.56 (1H, d, J ¼ 8.3 Hz), 8.76 (2H, br s), 8.97 (1H, s), 10.22
(1H, s); MS (FAB) m/z: 421 (MHþ); Anal. Calcd for
C25H28N2O4$HCl: C, 65.71; H, 6.40; N, 6.13; Cl, 7.76. Found: C,
65.68; H, 6.42; N, 6.06; Cl, 7.83.
The title compound was prepared in the same manner as
described for 2a using 6f instead of 6a as a colorless solid. 37% yield;
mp 206–207 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.85–3.05
(3H, m), 3.10–3.20 (3H, m), 3.83–3.87 (4H, m), 4.12 (1H, br s), 5.80
(1H, br s), 6.68 (2H, d, J ¼ 8.8 Hz), 6.77 (2H, d, J ¼ 8.8 Hz), 7.18 (1H, d,
J ¼ 8.3 Hz), 7.20–7.30 (1H, m), 7.39 (1H, d, J ¼ 7.8 Hz), 7.57 (2H, d,
J ¼ 8.3 Hz), 7.70–7.80 (1H, m), 8.50 (1H, d, J ¼ 4.9 Hz), 8.65 (1H, br s),
8.97 (1H, s), 10.27 (1H, s); MS (FAB) m/z: 422 (MHþ); Anal. Calcd for
C24H27N3O4$HCl$0.2H2O: C, 62.45; H, 6.20; N, 9.10; Cl, 7.68. Found:
C, 62.57; H, 6.18; N, 9.12; Cl, 7.54.
5.1.12. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2,2-diphenylacetanilide
hydrochloride (2b)
The title compound was prepared in the same manner as
described for 2a using 6b instead of 6a as a colorless powder. 88%
5.1.17. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(3-pyridyl)acetanilide
dihydrochloride (2g)
yield; 1H NMR (DMSO-d6)
d: 2.85–3.05 (3H, m), 3.05–3.20 (3H, m),
3.80–3.90 (2H, m), 4.05–4.20 (1H, m), 5.20 (1H, s), 5.82 (1H, d,
J ¼ 4.9 Hz), 6.68 (2H, d, J ¼ 8.8 Hz), 6.77 (2H, d, J ¼ 9.3 Hz), 7.19 (2H,
d, J ¼ 8.3 Hz), 7.20–7.50 (10H, m), 7.59 (1H, d, J ¼ 8.3 Hz), 8.77 (2H,
br s), 8.97 (1H, s), 10.50 (1H, s); MS (FAB) m/z: 497 (MHþ); Anal.
Calcd for C31H32N2O4$HCl: C, 69.85; H, 6.24; N, 5.26; Cl, 6.65.
Found: C, 69.66; H, 6.27; N, 5.02; Cl, 6.48.
The title compound was prepared in the same manner as
described for 2a using 6g instead of 6a as a colorless powder.
63% yield; 1H NMR (DMSO-d6)
d: 2.85–3.05 (3H, m), 3.10–3.25
(3H, m), 3.80–3.95 (2H, m), 3.99 (2H, s), 4.15–4.25 (1H, m),
6.65–6.80 (4H, m), 7.20 (2H, d, J ¼ 8.5 Hz), 7.58 (2H, d, J ¼ 8.5 Hz),
7.93–7.96 (1H, m), 8.43 (1H, d, J ¼ 8.0 Hz), 8.78–8.87 (3H, m), 9.10
(1H, br s), 10.61 (1H, s); MS (FAB) m/z: 422 (MHþ); Anal. Calcd for
C24H27N3O4$2HCl$1.8H2O: C, 54.72; H, 6.24; N, 7.98; Cl, 13.46.
Found: C, 54.81; H, 6.60; N, 7.82; Cl, 13.53.
5.1.13. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(1-naphthyl)acetanilide
hydrochloride (2c)
The title compound was prepared in the same manner as
described for 2a using 6c instead of 6a as a colorless solid. 43%
5.1.18. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(4-pyridyl)acetanilide
hydrochloride (2h)
yield; mp 226–228 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.85–
3.05 (3H, m), 3.10–3.20 (3H, m), 3.83–3.87 (2H, m), 4.10–4.20 (3H,
m), 5.80 (1H, br s), 6.68 (2H, d, J ¼ 8.8 Hz), 6.77 (2H, d, J ¼ 8.8 Hz),
7.18 (2H, d, J ¼ 8.8 Hz), 7.47–7.59 (6H, m), 7.85 (1H, d, J ¼ 7.3 Hz),
7.94 (1H, d, J ¼ 7.3 Hz), 8.13 (1H, d, J ¼ 7.8 Hz), 8.72 (2H, br s), 8.97
(1H, s), 10.39 (1H, s); MS (FAB) m/z: 471 (MHþ); Anal. Calcd for
The title compound was prepared in the same manner as
described for 2a using 6h instead of 6a as a colorless solid. 28%
yield; mp 228–229 ꢁC (MeOH–EtOH–EtOAc); 1H NMR (DMSO-d6)
d
: 2.91–3.03 (3H, m), 3.13–3.19 (3H, m), 3.71 (2H, s), 3.81–3.90
(2H, m), 4.14 (1H, br s), 5.81–5.83 (1H, m), 6.67–6.70 (2H, m),

T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
2539
6.75–6.78 (2H, m), 7.19 (2H, d, J ¼ 8.4 Hz), 7.33–7.35 (2H, m), 7.56
(2H, d, J ¼ 8.4 Hz), 8.50 (2H, d, J ¼ 6.0 Hz), 8.97 (1H, s), 10.34 (1H,
br s); MS (FAB) m/z: 422 (MHþ); Anal. Calcd for C24H27N3O4$HCl:
C, 62.95; H, 6.16; N, 9.18; Cl, 7.74. Found: C, 62.84; H, 6.28; N, 9.09;
Cl, 7.89.
2.65–2.80 (2H, m), 3.30–3.50 (4H, m), 3.48 (3H, s), 3.57 (2H, br s),
3.80–3.95 (2H, m), 4.05–4.15 (2H, m), 5.11 (2H, s), 6.61 (2H, d,
J ¼ 8.3 Hz), 6.82 (2H, d, J ¼ 9.3 Hz), 6.85–7.00 (2H, m), 6.97 (2H, d,
J ¼ 9.3 Hz); MS (EI) m/z: 446 (Mþ).
5.1.24. tert-Butyl (S)-N-{2-hydroxy-3-[4-
5.1.19. (S)-2-{[4-(Methoxymethoxy)phenoxy]methyl}oxirane (8a)
A mixture of 4-(methoxymethoxy)phenol (7a) (3.11 g), (R)-
epichlorohydrin (1.06 g) and KOH (1.1 g) in water (30 mL) was
stirred at room temperature for 15 h. The resultant mixture was
diluted with water and extracted with ethyl acetate. The organic
layer was washed with brine, and then dried and concentrated in
vacuo. The residue was purified using column chromatography on
silica gel with n-hexane/EtOAc (3:1) as the eluent to yield 8a
(methoxymethoxy)phenoxy]propyl}-N-(2-{4-[(2-hydroxy-2-
phenylacetyl)amino]phenyl}ethyl)carbamate (12a)
To a solution of 11 (0.47 g) and DL-mandelic acid (0.18 g) in N,N-
dimethylformamide (15 mL) were added 1-ethyl-3-(30-dimethyla-
minopropyl)carbodiimide hydrochloride (0.23 g) and 1-hydrox-
ybenzotriazole (0.16 g), and the mixture was stirred at room
temperature for 15 h. The resulting mixture was diluted with ethyl
acetate and washed with water twice. The organic layer was
washed with brine, and then dried and concentrated in vacuo. The
residue was purified using column chromatography on silica gel
with CHCl3/MeOH (30:1) as the eluent to yield 12a (0.61 g) as
(2.87 g) as a colorless oil. 67% yield; 1H NMR (CDCl3)
d: 2.73–2.75
(1H, m), 2.88–2.90 (1H, m), 3.31–3.35 (1H, m), 3.47 (3H, s), 3.90–
3.94 (1H, m), 4.11–4.19 (1H, m), 5.10 (2H, s), 6.85 (2H, d, J ¼ 9.2 Hz),
6.97 (2H, d, J ¼ 9.2 Hz); MS (EI) m/z: 210 (Mþ).
a yellow oil. 99% yield; 1H NMR (CDCl3)
d: 1.45 (9H, s), 2.75–2.90
(2H, m), 3.30–3.50 (4H, m), 3.47 (3H, s), 3.81–3.90 (2H, m), 4.04 (2H,
br s), 5.10 (2H, s), 5.19 (1H, d, J ¼ 3.4 Hz), 6.81 (2H, d, J ¼ 8.8 Hz), 6.96
(2H, d, J ¼ 8.8 Hz), 7.05–7.20 (2H, m), 7.35–7.49 (7H, m), 8.08 (1H, br
s); MS (FAB) m/z: 580 (Mþ).
5.1.20. (S)-2-{[4-(2-
Methoxyethoxymethoxy)phenoxy]methyl}oxirane (8b)
The title compound was prepared in the same manner as
described for 8a using 4-(2-methoxyethoxymethoxy)phenol (7b)
[16] instead of 7a as a colorless oil. 58% yield; 1H NMR (CDCl3)
d:
5.1.25. tert-Butyl (S)-N-(2-{4-[(2-tert-butoxycarbonylamino-2-
phenylacetyl)amino]phenyl}ethyl)-N-[2-hydroxy-3-[4-
(methoxymethoxy)phenoxy]propyl]carbamate (12b)
2.74 (1H, dd, J ¼ 4.8, 2.4 Hz), 2.89 (1H, t, J ¼ 40 Hz), 3.31–3.36 (1H,
m), 3.38 (3H, s), 3.54–3.58 (2H, m), 3.81–3.84 (2H, m), 3.92 (1H, dd,
J ¼ 5.2, 11.2 Hz), 4.17 (1H, dd, J ¼ 3.6, 10.4 Hz), 5.20 (2H, s), 6.82–6.87
(2H, m), 6.97–7.01 (2H, m); MS (EI) m/z: 254 (Mþ).
The title compound was prepared in the same manner as
described for 12a using N-Boc-phenylglycine instead of DL-man-
delic acid as a colorless oil. 82% yield; 1H NMR (CDCl3)
d: 1.41 (9H, s),
5.1.21. (S)-1-[4-(Methoxymethoxy)phenoxy]-3-[2-(4-
nitrophenyl)ethylamino]-2-propanol (9)
1.43 (9H, s), 2.70–2.90 (2H, m), 3.30–3.50 (4H, m), 3.47 (3H, s), 3.80–
3.95 (2H, m), 4.00–4.10 (2H, m), 5.10 (2H, s), 5.27 (1H, br s), 5.77
(1H, br s), 6.81 (2H, d, J ¼ 8.8 Hz), 6.96 (2H, d, J ¼ 9.3 Hz), 7.00–7.15
(2H, m), 7.33–7.44 (7H, m), 7.55 (1H, br s); MS (FAB) m/z: 679 (Mþ).
A mixture of 8a (10.16 g), 2-(4-nitrophenyl)ethylamine hydro-
chloride (9.6 g), and triethylamine (5.05 g) in 2-propanol (100 mL)
was refluxed for 8 h. After cooling to room temperature, the
resultant mixture was concentrated in vacuo and diluted with ethyl
acetate. The organic layer was washed with water and brine, and
then dried and concentrated in vacuo. The residue was purified
using column chromatography on silica gel with CHCl3/MeOH
(10:1) as the eluent to yield 9 (6.97 g) as a colorless oil. 38% yield;
5.1.26. tert-Butyl (S)-N-[2-(4-{[2-(2-
chlorophenyl)acetyl]amino}phenyl)ethyl]-N-{2-hydroxy-3-[4-
(methoxymethoxy)phenoxy]propyl}carbamate (12c)
To a solution of 11 (0.5 g) and triethylamine (0.19 g) in chloro-
form (15 mL) was added dropwise
a solution of 2-chlor-
1H NMR (CDCl3)
d: 2.77–3.00 (6H, m), 3.48 (3H, s), 3.92 (2H, d,
ophenylacetyl chloride (0.24 g) in benzene (2 mL), and the mixture
was stirred at room temperature for 2 h. The resulting mixture was
concentrated in vacuo, and the residue was purified using column
chromatography on silica gel with CHCl3/MeOH (30:1) as the eluent
J ¼ 4.9 Hz), 3.99–4.03 (1H, m), 5.11 (2H, s), 6.81 (2H, d, J ¼ 9.2 Hz),
6.96 (2H, d, J ¼ 8.6 Hz), 7.36 (2H, d, J ¼ 8.5 Hz), 8.15 (2H, d,
J ¼ 8.6 Hz); MS (FAB) m/z: 377 (MHþ).
to yield 12c (0.51 g) as a yellow oil. 81% yield; 1H NMR (CDCl3)
d:
5.1.22. tert-Butyl (S)-N-{2-hydroxy-3-[4-
(methoxymethoxy)phenoxy]propyl}-N-[2-(4-nitrophenyl)-
ethyl]carbamate (10)
1.44 (9H, s), 2.70–2.85 (2H, m), 3.30–3.50 (4H, m), 3.47 (3H, s),
3.75–3.95 (2H, m), 3.85 (2H, s), 4.00–4.15 (2H, m), 5.10 (2H, s), 6.80–
6.84 (2H, m), 6.91–7.00 (2H, m), 7.05–7.13 (2H, m), 7.26–7.52 (6H,
m); MS (FAB) m/z: 598 (MHþ).
To a solution of 9 (6.97 g) in tetrahydrofuran (80 mL) was
added di-tert-butyl dicarbonate (4.11 g). The mixture was stirred at
room temperature for 1 h, and then concentrated in vacuo. The
residue was purified using column chromatography on silica gel
with CHCl3/MeOH (30:1) as the eluent to yield 10 (8.45 g) as
5.1.27. tert-Butyl (S)-N-[2-(4-{[2-(3-
chlorophenyl)acetyl]amino}phenyl)ethyl]-N-{2-hydroxy-3-[4-
(methoxymethoxy)phenoxy]propyl}carbamate (12d)
The title compound was prepared in the same manner as
described for 12c using 3-chlorophenylacetyl chloride instead of 2-
chlorophenylacetyl chloride as a yellow oil. 98% yield; 1H NMR
a colorless oil. 96% yield; 1H NMR (CDCl3)
d: 1.43 (9H, s), 2.90–3.10
(2H, m), 3.40–3.60 (4H, m), 3.70 (3H, s), 3.80–3.95 (2H, m), 4.05–
4.15 (1H, m), 5.11 (2H, s), 6.82 (2H, d, J ¼ 8.8 Hz), 6.98 (2H, d,
J ¼ 9.3 Hz), 7.25–7.40 (2H, m), 8.15 (2H, d, J ¼ 8.3 Hz); MS (EI) m/z:
476 (Mþ).
(CDCl3) d: 1.45 (9H, s), 2.70–2.90 (2H, m), 3.30–3.50 (4H, m), 3.47
(3H, s), 3.69 (2H, s), 3.75–3.95 (2H, m), 4.00–4.20 (2H, m), 5.10 (2H,
s), 6.79–6.83 (2H, m), 6.91–7.00 (2H, m), 7.05–7.20 (2H, m), 7.21–
7.52 (6H, m); MS (FAB) m/z: 598 (MHþ).
5.1.23. tert-Butyl (S)-N-[2-(4-aminophenyl)ethyl]-N-{2-hydroxy-
3-[4-(methoxymethoxy)phenoxy]-propyl}carbamate (11)
To a solution of 10 (8.45 g) in ethanol (100 mL) was added
palladium on carbon (10% w/w, 0.5 g), and the mixture was stirred
under hydrogen atmosphere for 4 h. The catalyst was removed by
filtration, and the filtrate was concentrated in vacuo to yield 11
5.1.28. tert-Butyl (S)-N-[2-(4-{[2-(4-
chlorophenyl)acetyl]amino}phenyl)ethyl]-N-{2-hydroxy-3-[4-
(methoxymethoxy)phenoxy]propyl}carbamate (12e)
The title compound was prepared in the same manner as
described for 12c using 4-chlorophenylacetyl chloride instead of
(7.85 g) as a colorless oil. 99% yield; 1H NMR (CDCl3)
d: 1.46 (9H, s),

2540
T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
2-chlorophenylacetyl chloride as a yellow oil. 90% yield; 1H NMR
(CDCl3) : 1.45 (9H, s), 2.70–2.90 (2H, m), 3.30–3.50 (4H, m), 3.47
(3H, s), 3.69 (2H, s), 3.75–3.90 (2H, m), 4.00–4.15 (2H, m), 5.10 (2H,
s), 6.79–6.83 (2H, m), 6.90–7.20 (5H, m), 7.21–7.34 (5H, m); MS
(FAB) m/z: 598 (MHþ).
temperature for 1 h. The resulting mixture was concentrated in
vacuo, and the residue was solidified by trituration. The crude solid
was purified by recrystallization from EtOH–Et2O to yield 2i
(0.22 g) as a colorless solid. 44% yield; mp 173–176 ꢁC (EtOH–Et2O);
d
1H NMR (DMSO-d6)
d: 2.85–3.05 (3H, m), 3.10–3.20 (3H, m), 3.83–
3.87 (2H, m), 4.12 (1H, br s), 5.10 (1H, s), 5.80 (1H, br s), 6.68 (2H, d,
J ¼ 8.8 Hz), 6.77 (2H, d, J ¼ 9.3 Hz), 7.17 (2H, d, J ¼ 8.3 Hz), 7.28–7.37
(3H, m), 7.51 (2H, d, J ¼ 7.3 Hz), 7.66 (2H, d, J ¼ 8.3 Hz), 8.60–8.80
(2H, m), 8.97 (1H, br s), 9.93 (1H, s); MS (FAB) m/z: 437 (MHþ); Anal.
Calcd for C25H28N2O5$1.1HCl: C, 63.00; H, 6.15; N, 5.88; Cl, 8.18.
Found: C, 62.61; H, 6.16; N, 5.99; Cl, 8.01.
5.1.29. tert-Butyl (S)-N-{2-hydroxy-3-[4-
(methoxymethoxy)phenoxy]propyl}-N-[2-(4-{[2-(2-
methoxyphenyl)acetyl]amino}phenyl)ethyl]carbamate (12f)
The title compound was prepared in the same manner as
described for 12c using 2-methoxyphenylacetyl chloride instead of
2-chlorophenylacetyl chloride as a yellow oil. 72% yield; 1H NMR
(CDCl3)
d
: 1.44 (9H, s), 2.70–2.85 (2H, m), 3.30–3.50 (4H, m), 3.47
5.1.35. (S)-40-(2-{[2-Hydroxy-3-(4-
(3H, s), 3.70 (2H, s), 3.80–3.95 (2H, m), 3.92 (3H, s), 4.00–4.15 (2H,
m), 5.10 (2H, s), 6.81 (2H, d, J ¼ 8.8 Hz), 6.90–7.10 (6H, m), 7.20–7.34
(2H, m), 7.61 (1H, br s); MS (FAB) m/z: 595 (MHþ).
hydroxyphenoxy)propyl]amino}ethyl)-2-amino-2-
phenylacetanilide dihydrochloride (2j)
The title compound was prepared in the same manner as
described for 2i using 12b instead of 12a as a colorless powder. 80%
5.1.30. tert-Butyl (S)-N-{2-hydroxy-3-[4-
yield; 1H NMR (DMSO-d6)
d: 2.85–3.00 (3H, m), 3.10–3.20 (3H, m),
(methoxymethoxy)phenoxy]propyl}-N-[2-(4-{[2-(3-
3.80–3.90 (2H, m), 4.16 (1H, br s), 5.29 (1H, br s), 6.68 (2H, d,
J ¼ 9.3 Hz), 6.77 (2H, d, J ¼ 8.8 Hz), 7.21 (2H, d, J ¼ 8.8 Hz), 7.42–7.48
(3H, m), 7.59 (2H, d, J ¼ 8.8 Hz), 7.68 (2H, d, J ¼ 6.8 Hz), 8.80–9.10
(5H, m), 11.18 (1H, s); MS (FAB) m/z: 436 (MHþ); Anal. Calcd for
C25H29N3O4$2.2HCl$3.4H2O: C, 52.04; H, 6.64; N, 7.28; Cl, 13.52.
Found: C, 51.94; H, 6.98; N, 7.67; Cl, 13.55.
methoxyphenyl)acetyl]amino}phenyl)ethyl]carbamate (12g)
The title compound was prepared in the same manner as
described for 12c using 3-methoxyphenylacetyl chloride instead of 2-
chlorophenylacetylchlorideas ayellowoil. 86%yield; 1HNMR(CDCl3)
d: 1.45 (9H, s), 2.70–2.85 (2H, m), 3.30–3.50 (4H, m), 3.47 (3H, s), 3.70
(2H, s), 3.75–3.95 (2H, m), 3.82 (3H, s), 4.00–4.15 (2H, m), 5.10 (2H, s),
6.79–7.06 (10H, m), 7.26–7.34 (2H, m); MS (FAB) m/z: 594 (Mþ).
5.1.36. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(2-
5.1.31. tert-Butyl (S)-N-{2-hydroxy-3-[4-
chlorophenyl)acetanilide hydrochloride (2k)
(methoxymethoxy)phenoxy]propyl}-N-[2-(4-{[2-(4-
The title compound was prepared in the same manner as
described for 2i using 12c instead of 12a as a colorless solid. 43%
methoxyphenyl)acetyl]amino}phenyl)ethyl]carbamate (12h)
The title compound was prepared in the same manner as
described for 12c using 4-methoxyphenylacetyl chloride instead of
2-chlorophenylacetyl chloride as a yellow oil. 85% yield; 1H NMR
yield; mp 236–239 ꢁC (MeOH–Et2O); 1H NMR (DMSO-d6)
d: 2.85–
3.05 (3H, m), 3.10–3.20 (3H, m), 3.81–3.90 (2H, m), 3.83 (2H, s), 4.16
(1H, br s), 5.83 (1H, br s), 6.67–6.71 (2H, m), 6.75–6.78 (2H, m), 7.19
(2H, d, J ¼ 8.3 Hz), 7.27–7.33 (2H, m), 7.40–7.45 (2H, m), 7.57 (2H, d,
J ¼ 8.8 Hz), 8.78 (1H, br s), 8.98 (2H, br s), 10.29 (1H, s); MS (FAB) m/
z: 455 (MHþ); Anal. Calcd for C25H27N2O4Cl$HCl: C, 61.10; H, 5.74; N,
5.70; Cl, 14.43. Found: C, 60.92; H, 5.72; N, 5.60; Cl, 14.74.
(CDCl3) d: 1.45 (9H, s), 2.70–2.85 (2H, m), 3.30–3.50 (4H, m), 3.47
(3H, s), 3.67 (2H, s), 3.75–3.95 (2H, m), 3.83 (3H, s), 4.00–4.15 (2H,
m), 5.10 (2H, s), 6.79–6.84 (2H, m), 6.90–7.15 (6H, m), 7.23–7.26 (2H,
m), 7.32 (2H, d, J ¼ 8.3 Hz); MS (FAB) m/z: 594 (Mþ).
5.1.32. tert-Butyl (S)-N-{2-hydroxy-3-[4-
5.1.37. (S)-40-(2-{[2-Hydroxy-3-(4-
(methoxymethoxy)phenoxy]propyl}-N-[2-(4-{[2-(2-thenyl)-
acetyl]amino}phenyl)ethyl]carbamate (12i)
hydroxyphenoxy)propyl]amino}ethyl)-2-(3-
chlorophenyl)acetanilide hydrochloride (2l)
The title compound was prepared in the same manner as
described for 12c using 2-thenylacetyl chloride instead of 2-chlor-
ophenylacetyl chloride as a colorless oil. 74% yield; 1H NMR (CDCl3)
The title compound was prepared in the same manner as
described for 2i using 12d instead of 12a as a colorless solid. 31%
yield; mp 229–232 ꢁC (MeOH–Et2O); 1H NMR (DMSO-d6)
d: 2.85–
d
: 1.45 (9H, s), 2.70–2.85 (2H, m), 3.30–3.50 (4H, m), 3.48 (3H, s),
3.05 (3H, m), 3.10–3.20 (3H, m), 3.67 (2H, s), 3.81–3.90 (2H, m),
4.14 (1H, br s), 5.82 (1H, d, J ¼ 4.9 Hz), 6.67–6.71 (2H, m), 6.75–
6.78 (2H, m), 7.19 (2H, d, J ¼ 8.8 Hz), 7.28–7.41 (4H, m), 7.56 (2H, d,
J ¼ 8.3 Hz), 8.70 (1H, br s), 8.82 (1H, br s), 8.67 (1H, s), 10.28 (1H,
s); MS (FAB) m/z: 455 (MHþ); Anal. Calcd for C25H27N2O4Cl$HCl: C,
61.10; H, 5.74; N, 5.70; Cl, 14.43. Found: C, 61.23; H, 5.72; N, 5.69;
Cl, 14.58.
3.80–3.90 (2H, m), 3.94 (2H, s), 4.00–4.15 (2H, m), 5.10 (2H, s), 6.81
(2H, d, J ¼ 8.8 Hz), 6.97 (2H, d, J ¼ 8.8 Hz), 7.00–7.15 (3H, m), 7.20–
7.36 (4H, m); MS (FAB) m/z: 570 (Mþ).
5.1.33. tert-Butyl (S)-N-{2-hydroxy-3-[4-
(methoxymethoxy)phenoxy]propyl}-N-[2-(4-{[2-(3-thenyl)-
acetyl]amino}phenyl)ethyl]carbamate (12j)
The title compound was prepared in the same manner as
described for 12c using 3-thenylacetyl chloride instead of 2-chlor-
ophenylacetyl chloride as a colorless oil. 66% yield; 1H NMR (CDCl3)
5.1.38. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(4-
chlorophenyl)acetanilide hydrochloride (2m)
d
: 1.45 (9H, s), 2.70–2.85 (2H, m), 3.30–3.50 (4H, m), 3.48 (3H, s),
The title compound was prepared in the same manner as
described for 2i using 12e instead of 12a as a colorless solid. 49%
3.76 (2H, s), 3.80–3.90 (2H, m), 4.00–4.15 (2H, m), 5.10 (2H, s), 6.80–
6.83 (2H, m), 6.95–6.98 (2H, m), 7.06–7.08 (3H, m), 7.24–7.52 (4H,
m); MS (FAB) m/z: 571 (MHþ).
yield; mp 245–250 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.85–
3.05 (3H, m), 3.10–3.20 (3H, m), 3.65 (2H, s), 3.81–3.90 (2H, m), 4.13
(1H, br s), 5.82 (1H, br s), 6.68 (2H, d, J ¼ 8.8 Hz), 6.77 (2H, d,
J ¼ 8.8 Hz), 7.18 (2H, d, J ¼ 8.3 Hz), 7.33–7.40 (4H, m), 7.55 (2H, d,
J ¼ 8.3 Hz), 8.69 (1H, br s), 8.78 (1H, br s), 8.97 (1H, s), 10.25 (1H, s);
MS(FAB)m/z:455(MHþ);Anal. Calcd forC25H27N2O4Cl$HCl$0.2H2O:
C, 60.66; H, 5.78; N, 5.66; Cl, 14.32. Found: C, 60.53; H, 5.66; N, 5.55;
Cl, 14.68.
5.1.34. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-hydroxy-2-
phenylacetanilide hydrochloride (2i)
To a solution of 12a (0.6 g) in methanol (10 mL) was added 4 M
HCl–EtOAc solution (10 mL), and the mixture was stirred at room

T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
2541
5.1.39. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(2-
methoxyphenyl)acetanilide hydrochloride (2n)
8.96 (1H, s), 10.16 (1H, s); MS (FAB) m/z: 427 (MHþ); Anal. Calcd for
C23H26N2O4S$HCl: C, 59.67; H, 5.88; N, 6.05; S, 6.93; Cl, 7.66. Found:
C, 59.41; H, 5.91; N, 6.04; S, 6.79; Cl, 7.87.
The title compound was prepared in the same manner as
described for 2i using 12f instead of 12a as a colorless solid. 38%
5.1.44. 40-Cyanomethyl-2-(quinolin-2-yl)acetanilide (14a)
yield; mp 179–181 ꢁC (EtOH); 1H NMR (DMSO-d6)
d: 2.85–3.05 (3H,
A mixture of 4-aminobenzylcyanide (1.07 g) and ethyl 2-qui-
nolinylacetate (1.04 g) in xylene (15 mL) was refluxed for 19 h. After
cooling to room temperature, the resultant mixture was concen-
trated in vacuo. The residue was purified by recrystallization from
n-hexane–EtOAc to yield 14a (1.22 g) as a colorless solid. 79% yield;
m), 3.10–3.20 (3H, m), 3.61 (2H, s), 3.76 (3H, s), 3.83–3.87 (2H, m),
4.10–4.20 (1H, m), 5.82 (1H, d, J ¼ 4.4 Hz), 6.68 (2H, d, J ¼ 8.8 Hz),
6.77 (2H, d, J ¼ 8.8 Hz), 6.90 (1H, t, J ¼ 7.3 Hz), 6.98 (1H, d,
J ¼ 7.8 Hz), 7.16–7.24 (4H, m), 7.56 (2H, d, J ¼ 8.3 Hz), 8.65 (2H, br s),
8.96 (1H, s), 10.05 (1H, s); MS (FAB) m/z: 451 (MHþ); Anal. Calcd for
C26H30N2O5$HCl$0.1H2O: C, 63.89; H, 6.43; N, 5.73; Cl, 7.25. Found:
C, 63.77; H, 6.40; N, 5.68; Cl, 7.49.
1H NMR (CDCl3)
d: 3.71 (2H, s), 4.08 (2H, s), 7.25–7.30 (2H, m), 7.40
(1H, d, J ¼ 8.4 Hz), 7.57–7.66 (3H, m), 7.77–7.89 (2H, m), 8.12 (1H, d,
J ¼ 8.4 Hz), 8.20 (1H, d, J ¼ 8.4 Hz), 10.75 (1H, br s); MS (FAB) m/z:
302 (MHþ).
5.1.40. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(3-
methoxyphenyl)acetanilide hydrochloride (2o)
5.1.45. 40-Cyanomethyl-2-(isoquinolin-3-yl)acetanilide (14b)
The title compound was prepared in the same manner as
described for 14a using ethyl 3-isoquinolinylacetate [17] instead of
ethyl 2-quinolinylacetate as a colorless solid. 67% yield; 1H NMR
The title compound was prepared in the same manner as
described for 2i using 12g instead of 12a as a colorless solid. 43%
yield; mp 214–217 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.85–
(CDCl3) d: 3.70 (2H, s), 4.01 (2H, s), 7.23–7.28 (2H, m), 7.56–7.68 (4H,
3.05 (3H, m), 3.10–3.20 (3H, m), 3.59 (2H, s), 3.74 (3H, s), 3.81–3.90
(2H, m), 4.13 (1H, br s), 5.81 (1H, d, J ¼ 4.9 Hz), 6.68 (2H, d,
J ¼ 8.8 Hz), 6.75–6.91 (5H, m), 7.16–7.25 (3H, m), 7.56 (2H, d,
J ¼ 8.3 Hz), 8.67 (2H, br s), 8.96 (1H, s), 10.18 (1H, s); MS (FAB) m/z:
451 (MHþ); Anal. Calcd for C26H30N2O5$HCl: C, 64.13; H, 6.42; N,
5.75; Cl, 7.28. Found: C, 63.87; H, 6.34; N, 5.68; Cl, 7.61.
m), 7.71–7.76 (1H, m), 8.01 (1H, d, J ¼ 8.1 Hz), 9.29 (1H, s), 9.88 (1H,
br s); MS (FAB) m/z: 302 (MHþ).
5.1.46. 40-Cyanomethyl-2-(pyrimidin-2-yl)acetanilide (14c)
The title compound was prepared in the same manner as
described for 14a using ethyl 2-pyrimidinylacetate instead of ethyl
2-quinolinylacetate as a colorless solid. 89% yield; 1H NMR (CDCl3)
5.1.41. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(4-
methoxyphenyl)acetanilide hydrochloride (2p)
d: 3.72 (2H, s), 4.13 (2H, s), 7.26–7.31 (3H, m), 7.59–7.62 (2H, m),
8.78 (2H, d, J ¼ 5.2 Hz), 9.82 (1H, s); MS (FAB) m/z: 253 (MHþ).
The title compound was prepared in the same manner as
described for 2i using 12h instead of 12a as a colorless solid. 52%
5.1.47. 40-Cyanomethyl-2-(pyrazin-2-yl)acetanilide (14d)
The title compound was prepared in the same manner as
described for 14a using ethyl 2-pyrazinylacetate [18] instead of
ethyl 2-quinolinylacetate as a colorless solid. 76% yield; 1H NMR
yield; mp 241–244 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.85–
3.05 (3H, m), 3.10–3.20 (3H, m), 3.55 (2H, s), 3.72 (3H, s), 3.81–3.90
(2H, m), 4.16 (1H, br s), 5.84 (1H, br s), 6.69 (2H, d, J ¼ 8.8 Hz), 6.77
(2H, d, J ¼ 8.8 Hz), 6.88 (2H, d, J ¼ 8.8 Hz), 7.17 (2H, d, J ¼ 8.3 Hz),
7.25 (2H, d, J ¼ 8.8 Hz), 7.56 (2H, d, J ¼ 8.3 Hz), 8.77 (1H, br s), 8.98
(2H, br s), 10.21 (1H, s); MS (FAB) m/z: 451 (MHþ); Anal. Calcd for
C26H30N2O5$HCl$0.1H2O: C, 63.89; H, 6.43; N, 5.73; Cl, 7.25. Found:
C, 63.65; H, 6.31; N, 5.62; Cl, 7.46.
(CDCl3) d: 3.71 (2H, s), 3.92 (2H, s), 7.26–7.29 (2H, m), 7.55–7.58 (2H,
m), 8.57–8.60 (2H, m), 8.65 (1H, s), 9.12 (1H, s); MS (FAB) m/z: 253
(MHþ).
5.1.48. (S)-40-[2-({2-Hydroxy-3-[4-(2-
methoxyethoxymethoxy)phenoxy]propyl}amino)ethyl]-2-(quinolin-
2-yl)acetanilide (15a)
5.1.42. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(2-thenyl)acetanilide
hydrochloride (2q)
To a mixture of 14a (2.09 g) and concentrated aqueous ammonia
solution (1 mL) in tetrahydrofuran (40 mL) was added Raney-
nickel, and the mixture was stirred under hydrogen atmosphere for
4 h. The catalyst was removed by filtration over Celite, and the
filtrate was concentrated in vacuo. The residue was added to
a solution of 8b (1.44 g) in 2-propanol (40 mL), and the mixture was
refluxed for 1 h. After cooling to room temperature, the resultant
mixture was concentrated in vacuo. The residue was purified using
column chromatography on silica gel with CHCl3/MeOH (20:1) as
the eluent to yield 15a (1.27 g) as a colorless powder. 32% yield; 1H
The title compound was prepared in the same manner as
described for 2i using 12i instead of 12a as a colorless solid. 55%
yield; mp 219–223 ꢁC (MeOH–Et2O); 1H NMR (DMSO-d6)
d: 2.89–
3.02 (3H, m), 3.16 (3H, br s), 3.81–3.90 (2H, m), 3.87 (2H, s), 4.14
(1H, br s), 5.83 (1H, br s), 6.68 (2H, d, J ¼ 8.8 Hz), 6.77 (2H, d,
J ¼ 8.8 Hz), 6.96–6.98 (2H, m), 7.19 (2H, d, J ¼ 8.3 Hz), 7.37–7.39 (1H,
m), 7.56 (2H, d, J ¼ 7.8 Hz), 8.72 (1H, br s), 8.85 (1H, br s), 8.97 (1H,
s), 10.27 (1H, s); MS (FAB) m/z: 427 (MHþ); Anal. Calcd for
C23H26N2O4S$HCl$0.1H2O: C, 59.44; H, 5.90; N, 6.03; S, 6.90; Cl,
7.63. Found: C, 59.33; H, 5.85; N, 6.01; S, 6.77; Cl, 7.89.
NMR (CDCl3) d: 2.70–2.93 (6H, m), 3.37 (3H, s), 3.54–3.57 (2H, m),
3.79–3.84 (2H, m), 3.88–3.93 (2H, m), 3.95–4.04 (1H, m), 4.06 (2H,
s), 5.18 (2H, s), 6.77–6.85 (2H, m), 6.93–7.00 (2H, m), 7.15 (2H, d,
J ¼ 4.0 Hz), 7.41 (1H, d, J ¼ 8.4 Hz), 7.50–7.60 (3H, m), 7.75–7.87 (2H,
m), 8.11 (1H, d, J ¼ 8.4 Hz), 8.17 (1H, d, J ¼ 8.0 Hz), 10.42 (1H, br s);
MS (FAB) m/z: 560 (MHþ).
5.1.43. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(3-thenyl)acetanilide
hydrochloride (2r)
The title compound was prepared in the same manner as
described for 2i using 12j instead of 12a as a colorless solid. 32%
5.1.49. (S)-40-[2-({2-Hydroxy-3-[4-(2-
methoxyethoxymethoxy)phenoxy]propyl}amino)ethyl]-2-
(isoquinolin-3-yl)acetanilide (15b)
yield; mp 233–235 ꢁC (MeOH); 1H NMR (DMSO-d6)
d: 2.89–3.02
(3H, m), 3.16 (3H, br s), 3.65 (2H, s), 3.81–3.90 (2H, m), 4.13 (1H, br
s), 5.82 (1H, d, J ¼ 4.8 Hz), 6.67–6.70 (2H, m), 6.75–6.78 (2H, m),
7.07 (1H, d, J ¼ 5.2 Hz), 7.18 (2H, d, J ¼ 8.4 Hz), 7.31 (1H, d,
J ¼ 2.0 Hz), 7.46–7.49 (1H, m), 7.56 (2H, d, J ¼ 8.4 Hz), 8.67 (2H, br s),
The title compound was prepared in the same manner as
described for 15a using 14b instead of 14a as a colorless powder.
44% yield; 1H NMR (CDCl3)
d: 2.70–2.95 (6H, m), 3.37 (3H, s), 3.53–
3.57 (2H, m), 3.79–3.88 (2H, m), 3.87–3.92 (2H, m), 3.95–4.03 (3H,

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T. Maruyama et al. / European Journal of Medicinal Chemistry 44 (2009) 2533–2543
m), 5.19 (2H, s), 6.78–6.83 (2H, m), 6.94–7.00 (2H, m), 7.13 (2H, d,
J ¼ 8.4 Hz), 7.47 (2H, d, J ¼ 8.4 Hz), 7.59–7.75 (4H, m), 7.99 (1H, d,
J ¼ 7.9 Hz), 9.28 (1H, s), 9.62 (1H, br s); MS (FAB) m/z: 560 (MHþ).
J ¼ 8.8 Hz), 8.69 (2H, br), 8.77 (2H, d, J ¼ 5.2 Hz), 8.98 (1H, s), 10.29
(1H, s); MS (FAB) m/z: 423 (MHþ); Anal. Calcd for
C23H26N4O4$1.1HCl: C, 59.72; H, 5.90; N, 12.11; Cl, 8.43. Found: C,
59.49; H, 5.87; N, 11.83; Cl, 8.13.
5.1.50. (S)-40-[2-({2-Hydroxy-3-[4-(2-
methoxyethoxymethoxy)phenoxy]propyl}amino)ethyl]-2-
(pyrimidin-2-yl)acetanilide (15c)
5.1.55. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(pyrazin-2-yl)acetanilide
hydrochloride (2v)
The title compound was prepared in the same manner as
described for 15a using 14c instead of 14a as a colorless powder.
The title compound was prepared in the same manner as
described for 2i using 15d instead of 12a as a colorless solid. 79%
24% yield; 1H NMR (CDCl3)
d: 2.78–2.85 (3H, m), 2.90–2.99 (3H, m),
3.37 (3H, s), 3.54–3.57 (2H, m), 3.80–3.83 (2H, m), 3.90–3.92 (2H,
m), 4.04–4.08 (1H, m), 4.12 (2H, s), 5.19 (2H, s), 6.78–6.82 (2H, m),
6.95–6.98 (2H, m), 7.17 (2H, d, J ¼ 8.4 Hz), 7.23–7.29 (1H, m), 7.50
(2H, d, J ¼ 8.0 Hz), 8.76–8.78 (2H, m), 9.61 (1H, s); MS (FAB) m/z: 511
(MHþ).
yield; mp 218–219 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.88–
3.02 (3H, m), 3.16 (3H, br), 3.81–3.90 (2H, m), 3.94 (2H, s), 4.15 (1H,
br), 5.82–5.84 (1H, m), 6.66–6.71 (2H, m), 6.75–6.78 (2H, m), 7.19
(2H, d, J ¼ 8.4 Hz), 7.57 (2H, d, J ¼ 8.0 Hz), 8.54 (1H, d, J ¼ 2.8 Hz),
8.58 (1H, br s), 8.67 (1H, s), 8.74 (1H, br s), 8.89 (1H, br s), 8.98 (1H,
s), 10.38 (1H, s); MS (FAB) m/z: 423 (MHþ); Anal. Calcd for
C23H26N4O4$HCl$0.2H2O: C, 59.72; H, 5.97; N, 12.11; Cl, 7.66. Found:
C, 59.59; H, 5.91; N, 12.06; Cl, 7.97.
5.1.51. (S)-40-[2-({2-Hydroxy-3-[4-(2-
methoxyethoxymethoxy)phenoxy]propyl}amino)ethyl]-2-(pyrazin-
2-yl)acetanilide (15d)
The title compound was prepared in the same manner as
described for 15a using 14d instead of 14a as a colorless powder.
5.2. Pharmacology
33% yield; 1H NMR (CDCl3)
d: 2.78–2.85 (3H, m), 2.91–2.99 (3H, m),
5.2.1. Agonistic activity on human
Human 3-, 2-, and 1-stimulating activities were investigated
using a CHO cell system (cells in which human 3-, 2-, and 1-ARs
b3-, b2-, and b1-ARs
3.37 (3H, s), 3.54–3.57 (2H, m), 3.80–3.83 (2H, m), 3.90–3.92 (2H,
m), 4.04–4.08 (1H, m), 4.12 (2H, s), 5.19 (2H, s), 6.78–6.82 (2H, m),
6.95–6.98 (2H, m), 7.17 (2H, d, J ¼ 8.4 Hz), 7.23–7.29 (1H, m), 7.51
(2H, d, J ¼ 8.0 Hz), 8.76–8.78 (2H, m), 9.61 (1H, s); MS (FAB) m/z: 511
(MHþ).
b
b
b
b
b
b
are compulsorily expressed were used). The agonistic activity of the
compound (final concentrations of 10ꢂ10 to 10ꢂ4 M) was investi-
gated by incubating 105 cells/well of each the cells on a 24-well
plate and checking the activity after 2 days’ incubation (subcon-
fluent state) using the production of cyclic AMP (cAMP) as an index.
The amount of cAMP produced in each cell (pmol/ml) was
measured using a radioimmunoassay method with 125I-cAMP. The
intensity of action among compounds was compared by calculating
the EC50 and intrinsic activity (IA where the maximum reaction of
10ꢂ4 M isoproterenol was defined as 1.00) for each from the
resulting dose–reaction curve.
5.1.52. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(quinolin-2-yl)acetanilide
hydrochloride (2s)
The title compound was prepared in the same manner as
described for 2i using 15a instead of 12a as a colorless solid. 51%
yield; mp 226–227 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.87–
3.02 (3H, m), 3.16 (3H, br s), 3.81–3.90 (2H, m), 4.01 (2H, s), 4.16
(1H, br s), 5.84–5.85 (1H, m), 6.67–6.71 (2H, m), 6.75–6.78 (2H, m),
7.20 (2H, d, J ¼ 8.4 Hz), 7.57–7.61 (4H, m), 7.73–7.77 (1H, m), 7.97
(2H, d, J ¼ 8.8 Hz), 8.34 (1H, d, J ¼ 8.4 Hz), 8.77 (1H, br s), 8.93 (1H,
br s), 8.99 (1H, s), 10.45 (1H, s); MS (FAB) m/z: 472 (MHþ); Anal.
Calcd for C28H29N3O4$HCl: C, 66.20; H, 5.95; N, 8.27; Cl, 6.98.
Found: C, 65.96; H, 5.98; N, 8.27; Cl, 7.10.
5.2.2. Hypoglycemic activity in kk mice
Male kk mice (blood sugar level: not lower than 200 mg/dl)
were subjected to measurement of blood sugar level under fed
conditions, and then randomly classified into groups. The test
compound was administered orally once daily for 4 days, and the
blood sugar level 15–18 h after final administration was compared
with that before administration (n ¼ 6). Blood samples were
collected from the tail vein using a glass capillary tube (previously
treated with heparin) after which the blood was deproteinized,
and the amount of glucose in the supernatant (mg/dl) was
determined calorimetrically by means of the glucose oxidase
method.
5.1.53. (S)-40-(2-{[2-Hydroxy-3-(4-
hydroxyphenoxy)propyl]amino}ethyl)-2-(isoquinolin-3-
yl)acetanilide hydrochloride (2t)
The title compound was prepared in the same manner as
described for 2i using 15b instead of 12a as a colorless solid. 56%
yield; mp 214–216 ꢁC (MeOH–EtOH); 1H NMR (DMSO-d6)
d: 2.90–
3.02 (3H, m), 3.16 (3H, br s), 3.81–3.90 (2H, m), 4.00 (2H, s), 4.15
(1H, br s), 5.83–5.84 (1H, m), 6.66–6.71 (2H, m), 6.76–6.78 (2H, m),
7.19 (2H, d, J ¼ 8.8 Hz), 7.59–7.68 (3H, m), 7.75–7.80 (2H, m), 7.95
(1H, d, J ¼ 8.4 Hz), 8.12 (1H, d, J ¼ 8.0 Hz), 8.73 (1H, br s), 8.86 (1H,
br s), 8.98 (1H, s), 9.27 (1H, s), 10.33 (1H, s); MS (FAB) m/z: 472
(MHþ); Anal. Calcd for C28H29N3O4$HCl$0.1H2O: C, 65.97; H, 5.97;
N, 8.24; Cl, 6.95. Found: C, 65.83; H, 5.90; N, 8.23; Cl, 6.82.
Acknowledgments
The authors are grateful to the staff of the Division of Analytical
Science Laboratories for elemental analysis and spectral
measurements.
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5.1.54. (S)-40-(2-{[2-Hydroxy-3-(4-
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