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

Article abstract of DOI:10.1016/j.bmc.2009.06.031

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, we prepared a novel series of phenoxypropanolamine derivatives containing the th

Full text of DOI:10.1016/j.bmc.2009.06.031

Bioorganic & Medicinal Chemistry 17 (2009) 5510–5519
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Discovery of novel thiourea derivatives as potent and selective
b3-adrenergic receptor agonists
*
Tatsuya Maruyama , Norio Seki, Kenichi Onda, Takayuki Suzuki, Souichirou Kawazoe, 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
the treatment of obesity and noninsulin-dependent (type II) diabetes, we prepared a novel series of phen-
oxypropanolamine derivatives containing the thiourea moiety and evaluated their biological activities at
human b3-, b2-, and b1-ARs. Among these compounds, 4-nitrophenylthiourea (18i) and 3-methoxyphen-
ylthiourea (18k) derivatives were found to exhibit potent agonistic activity at the b3-AR, with EC50 values
Received 1 May 2009
Revised 15 June 2009
Accepted 16 June 2009
Available online 21 June 2009
of 0.10 and 0.16
showed significant hypoglycemic activity in a rodent diabetic model.
Ó 2009 Elsevier Ltd. All rights reserved.
lM, respectively, and no agonistic activity for either the b1- or b2-AR. In addition, they
Keywords:
b3-Adrenergic receptor
Agonist
Thiourea
Phenoxypropanolamine
Diabetes
1. Introduction
have been disappointing due to a lack of selectivity and/or potency
or poor pharmacokinetics.14,15
In 1984, b3-adrenergic receptors (ARs) were proposed as a third
group of b-ARs present in rat adipose tissue that utilize various
b-agonists.1,2 In 1989, the primary structure of the b3-AR was
identified and characterized using cloning and molecular pharma-
cological techniques.3–5 b3-AR was found to play a significant role
in regulating lipolysis and thermogenesis in both rodent and hu-
man adipose tissue. These findings suggest that b3-AR agonists
are suitable for mediating thermogenesis for the purpose of obesity
modulation and lowering plasma glucose and insulin levels, there-
by ameliorating noninsulin-dependent (type II) diabetes. Recent
studies indicated that, in addition to adipocytes, the b3-AR is also
distributed in human urinary bladder detrusor tissue and its relax-
ation occurs mainly via b3-AR.6–8 Subtype selectivity for b3-AR
agonists must be kept specifically in mind since activation of the
b1- or b2-ARs would cause undesirable side effects such as in-
creased heart rate or muscle tremors. A number of laboratories
have been engaged in developing potent and selective b3-AR ago-
nists for the treatment of obesity and type II diabetes.9,10 While
early potent and selective rat b3-AR agonists, such as BRL-
3734411 and CL-316243,12 were reported to be effective anti-obes-
ity and anti-diabetic agents in rodents (Fig. 1),13 clinical trials with
these drugs for the treatment of metabolic disorders in humans
In 1998, researchers at Merck reported a 4-hydroxyphenoxypro-
panolamine derivative containing the benzenesulfonamide moiety
(1) as one of the first selective human b3-AR agonists.16 Owing to
its rapid glucuronidation, however, the pharmacokinetic profile of
this agent was poor.17 Considering the structure of compound 1,
we assumed that the 4-hydroxyphenoxy moiety on the left-hand
side of 1 might be easily metabolized. In fact, we found that the oral
bioavailability of compound 1 in rats (5 mg/kg iv, 30 mg/kg po) was
low (F = 4%). In contrast, its phenoxy analogue 218 showed improved
oral bioavailability in rats (F = 44%). In addition, compound 2 exhib-
ited only a fivefold loss in potency at the b3-AR (EC50 = 0.41
IA = 0.89) compared to 1 (EC50 = 0.078 M, IA = 0.73).
lM,
l
Since compound 2 showed potent b3-AR agonistic activity and
good pharmacokinetic profile, we chose compound 2 as our lead
compound. Based on the structure of compound 2, we decided to
explore alternative structures instead of the sulfonamide moiety
of 2, and subsequently identified the thiourea derivative 14b.
We then further attempted to modify of the substituent on the
OH
H
OH
H
O
N
Cl
N
Cl
O
O
ONa
ONa
OH
O
O
O
CL-316243
BRL-37344
* Corresponding author. Tel.: +81 29 863 6749; fax: +81 29 852 5387.
E-mail address: tatsuya.maruyama@jp.astellas.com (T. Maruyama).
Figure 1. Chemical structure of early b3-AR agonists.
0968-0896/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2009.06.031

T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
5511
OH
H
OH
H
N
O
N
O
O O
S
H
N
X
A
1 (X = OH)
2 (X = H)
OH
H
O
N
S
R
N
N
H
H
Figure 2. Design of compounds.
thiourea moiety (Fig. 2). In this paper, we describe the synthesis
and structure–activity relationships (SARs) of these newly de-
signed phenoxypropanolamine derivatives as b3-AR agonists.
was coupled with the appropriate isothiocyanates, followed by
deprotection of Boc group to afford the desired products 18a–r.
3. Results and discussion
2. Chemistry
The prepared compounds were evaluated for their agonistic
activities in stimulating an increase in cyclic AMP (cAMP) levels
in Chinese hamster ovary (CHO) cells expressing cloned human
b3-, b2-, and b1-ARs. The results for the reference compounds, iso-
proterenol (ISO; non-selective b-AR agonist) and compound 1, are
also shown in Table 1 for comparison.
Compound 2 showed b3-, b2-, and b1-AR agonistic activity with
EC50 values of 0.41, >100, and 78 lM, respectively. In order to im-
prove b3-AR agonistic activity and functional selectivity over b1-
AR, modification of the sulfonamido moiety in 2 with the other
functional moieties containing the NH part was investigated (Table
1). Initially, reverse sulfonamide derivative 4 showed a fivefold de-
Reverse sulfonamide derivative 4 and aniline derivative 9 were
prepared as illustrated in Scheme 1. A commercially available N-
phenyl-4-(2-aminoethyl)benzenesulfonamide (3) was coupled
with (S)-2-(phenoxymethyl)oxirane to afford the desired product
4. N-Benzyl-4-aminophenylacetamide (5)19 was treated with tri-
phenylbismuth in the presence of copper diacetate, followed by
reduction of the amido moiety to give the 2-phenylethylamine 7.
Compound 7 was coupled with (S)-2-(phenoxymethyl)oxirane, fol-
lowed by deprotection of the benzyl group to afford the desired
product 9.
Compounds 12 and 14 were prepared from aniline intermediate
1020 as illustrated in Scheme 2. The coupling of 10 with benzoyl
chloride or phenyl chloroformate in the presence of triethylamine,
followed by deprotection of the tert-butoxycarbonyl (Boc) group
afforded the desired products 12a,b. Compound 10 was coupled
with phenylisocyanate or phenylisothiocyanate, followed by
deprotection of the Boc group to afford the desired products 14a,b.
Scheme 3 shows the synthesis of cyanoguanidine derivative 16.
The thiourea moiety of 13b was converted to a cyanoguanidine
moiety by treatment with cyanamide and 1-ethyl-3-(30-dimethyl-
aminopropyl)carbodiimide hydrochloride, followed by deprotec-
tion of the Boc group to afford the desired product 16.
crease in potency at the b3-AR (EC50 = 2.0
lM) and a dramatic in-
crease in potency at the b1-AR (EC50 = 0.25
lM) relative to 2.
These results indicated that the NH position is important for b3-
AR agonistic activity and selectivity over b1-AR. Aniline derivative
9 was 30-fold less potent at the b3-AR (EC50 = 12 lM) and dis-
played a decrease in intrinsic activity (IA = 0.31) relative to 2. This
revealed that the basic aniline group may not be efficacious in
improving agonistic activity at the b3-AR. Replacement of the sul-
fonamido moiety of 2 with the amido moiety (12a) resulted in dra-
matically decreased potency at the b3-AR (EC50 = 130
urethane derivative 12b exhibited 23-fold decreased potency at
the b3-AR (EC50 = 9.3 M) relative to 2. Next, replacement of the
sulfonamido moiety in 2 with the ureido moiety (14a) resulted
lM), while
Thiourea derivatives 18a–r were prepared in the same manner
as the synthesis of 14b as illustrated in Scheme 4. Compound 10
l
OH
H
H2N
O
N
a
H
N
O O
H
N
O O
S
S
4
3
b
BnHN
c
BnHN
BnHN
d
O
e
O
N
H
NH2
N
H
7
5
6
OH
OH Bn
N
H
N
O
O
N
H
N
9
H
8
Scheme 1. Reagents and conditions: (a) (S)-2-(phenoxymethyl)oxirane, MeCN, reflux, and then 4 M HCl–dioxane, MeOH; (b) Ph3Bi, Cu(OAc)2, CH2Cl2, reflux; (c) 1 M BH3–
i
THF, THF, reflux; (d) (S)-2-(phenoxymethyl)oxirane, PrOH, dioxane, reflux; (e) H2, Pd/C, EtOH, and then 4 M HCl–EtOAc, MeOH.

5512
T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
OH
OH Boc
H
OH Boc
N
O
N
O
N
a
b
O
O
O
R
N
N
R
NH2
H
H
12a,b
11a,b
10
12a : R = Ph
12b : R = OPh
11a : R = Ph
11b : R = OPh
OH Boc
N
OH
H
N
O
O
c
b
X
X
N
N
N
N
H
H
H
H
13a,b
14a,b
13a : X = O
13b : X = S
14a : X = O
14b : X = S
Scheme 2. Reagents and conditions: (a) RCOCl, Et3N, CHCl3; (b) 4 M HCl–EtOAc, EtOH; (c) PhCNX, CHCl3, 60–80 °C.
OH
H
OH Boc
N
CN
CN
O
N
O
b
N
a
N
13b
N
N
N
N
H
H
H
H
16
15
i
Scheme 3. Reagents and conditions: (a) H2NCN, EDCꢀHCl, Pr2NEt, DMF, 50 °C; (b) CF3CO2H, and then 4 M HCl–dioxane.
OH Boc
N
OH
H
O
O
N
b
a
S
S
10
R
R
N
N
N
N
H
H
H
H
17a-r
18a-r
17a : R = cyclohexyl
17b : R = benzyl
18a : R = cyclohexyl
18b : R = benzyl
17c : R = 2-phenylethyl
18c : R = 2-phenylethyl
17d : R = 2-chlorophenyl
17e : R = 3-chlorophenyl
17f : R = 4-chlorophenyl
17g : R = 2-nitrophenyl
17h : R = 3-nitrophenyl
17i : R = 4-nitrophenyl
18d : R = 2-chlorophenyl
18e : R = 3-chlorophenyl
18f : R = 4-chlorophenyl
18g : R = 2-nitrophenyl
18h : R = 3-nitrophenyl
18i : R = 4-nitrophenyl
17j : R = 2-methoxyphenyl
17k : R = 3-methoxyphenyl
17l : R = 4-methoxyphenyl
17m : R = 4-fluorophenyl
17n : R = 4-bromophenyl
17o : R = 4-iodophenyl
18j : R = 2-methoxyphenyl
18k : R = 3-methoxyphenyl
18l : R = 4-methoxyphenyl
18m : R = 4-fluorophenyl
18n : R = 4-bromophenyl
18o : R = 4-iodophenyl
17p : R = 4-methoxycarbonylphenyl
17q : R = 4-sulfamoylphenyl
17r : R = 4-methylphenyl
18p : R = 4-methoxycarbonylphenyl
18q : R = 4-sulfamoylphenyl
18r : R = 4-methylphenyl
Scheme 4. Reagents and conditions: (a) RNCS, CHCl3, 80 °C; (b) 4 M HCl–EtOAc, EtOH.
in a fivefold decrease in potency at the b3-AR with partial agonistic
activity (EC50 = 1.9 M, IA = 0.38) relative to 2. In contrast, thiourea
derivative 14b showed comparable agonistic activity at the b3-AR
(EC50 = 0.44 M, IA = 0.70) relative to 2, and no agonistic activity
for either the b1- or b2-AR. These results indicated that the thio-
urea group played an important role in b3-AR agonistic activity
and selectivity over b1-AR. However, cyanoguanidine derivative
16, considered a bioisostere of thiourea, showed a 75-fold loss in
or ethylene group between the phenyl ring and thiourea moiety in
14b may not have been efficacious for improving agonistic activity
at the b3-AR.
We then examined the effects of substituents on the phenyl ring
of the thiourea moiety in 14b as shown in Table 3. The introduction
of a chloro group as an electron-withdrawing group on the phenyl
ring of the thiourea moiety in 14b (18d–f) resulted in slightly in-
creased potency at the b3-AR relative to 14b, with an apparent
rank order of potency of para- and meta-chloro (EC50 = 0.28 and
l
l
potency at the b3-AR (EC50 = 33 lM) relative to 14b. We therefore
selected the thiourea derivative 14b as our new leading candidate.
In order to improve b3-AR agonistic activity, modification of the
substituent on the thiourea moiety in 14b was examined as shown
in Table 2. Replacement of the phenyl ring with a cyclohexyl ring
(18a) resulted in threefold decreased potency at the b3-AR
0.27 lM) > ortho-chloro (EC50 = 0.40 lM). The nitro group as a
strong electron-withdrawing group was also introduced on the
phenyl ring of the thiourea moiety (18g–i) to yield improved po-
tency of the b3-AR relative to 14b, with a similar rank order of po-
tency to those of the chlorophenyl derivatives. When the intrinsic
activities of b3-AR for 2-nitrophenyl (18g) and 3-nitrophenyl (18h)
derivatives were partial, 4-nitrophenyl derivative 18i showed full
b3-AR agonistic activity (EC50 = 0.10 lM, IA = 0.88), and was the
most potent in this series. The introduction of a methoxy group
as an electron-donating group on the phenyl ring of the thiourea
(EC50 = 1.4 lM) relative to 14b. Both the benzyl (18b) and 2-phen-
ylethyl (18c) derivatives exhibited maintained potency at the b3-
AR, but their intrinsic activities were decreased (IA = 0.53 and
0.34, respectively). These results revealed that replacement of the
phenyl ring with a cyclohexyl ring and the insertion of a methylene

T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
5513
Table 1
b-AR agonistic activity of phenoxypropanolamine derivatives
Table 3
b-AR agonistic activity of substituted phenylthiourea derivatives
OH
OH
H
H
N
3
4
N
O
O
2
S
R
A
N
H
N
H
Compound
A
EC50 (l
M) (IAb)
a
a
Compound
R
EC50 (l
M) (IAb)
b3-AR
b2-AR
b1-AR
b3-AR
b2-AR
b1-AR
2
4
9
12a
12b
14a
14b
16
1
NHSO2
SO2NH
NH
NHCO
0.41 (0.89)
2.0 (0.52)
12 (0.31)
130 (0.33)
9.3 (0.57)
1.9 (0.38)
0.44 (0.70)
33 (0.55)
>100 (0)
>100 (0.05)
>100 (0)
>100 (0)
>100 (0)
>100 (0)
>100 (0)
>100 (0.03)
>100 (0.02)
78 (0.11)
14b
18d
18e
18f
18g
18h
18i
18j
18k
18l
18m
18n
18o
18p
18q
18r
H
2-Cl
3-Cl
4-Cl
2-NO2
3-NO2
4-NO2
2-OMe
3-OMe
4-OMe
4-F
4-Br
4-I
4-CO2Me
4-SO2NH2
4-Me
0.44 (0.70)
0.40 (0.61)
0.27 (0.61)
0.28 (0.55)
0.34 (0.45)
0.10 (0.45)
0.10 (0.88)
0.27 (0.76)
0.16 (0.81)
0.20 (0.63)
0.23 (0.58)
0.23 (0.46)
0.47 (0.57)
0.17 (0.37)
0.32 (0.49)
0.65 (0.60)
>100 (0)
>100 (0.11)
>100 (0.06)
>100 (0.10)
>100 (0.03)
>100 (0.06)
>100 (0.08)
>100 (0.09)
0.34 (0.13)
>100 (0.07)
>100 (0.07)
>100 (0.09)
>100 (0.07)
>100 (0.07)
>100 (0.06)
>100 (0.03)
>100 (0.10)
0.25 (0.39)
>100 (0.01)
>100 (0.02)
>100 (0.04)
>100 (0.02)
>100 (0.11)
0.056 (0.14)
1.6 (0.21)
>100 (0.04)
>100 (0.05)
>100 (0.03)
>100 (0.02)
>100 (0.03)
>100 (0.02)
>100 (0.01)
>100 (0.02)
>100 (0.02)
>100 (0.01)
>100 (0.02)
>100 (0.01)
>100 (0.01)
>100 (0.01)
>100 (0.01)
NHCO2
NHCONH
NHCSNH
NHC(NCN)NH
0.078 (0.73)
ISO
0.10 (1.00)
0.003 (1.00)
0.012 (1.00)
a
Agonistic activity was assessed by measuring cAMP accumulation in CHO cells
expressing b-ARs.
b
Values in parentheses represent the intrinsic activity (IA) given as a fraction of
the maximum stimulation with isoproterenol.
a
Agonistic activity was assessed by measuring cAMP accumulation in CHO cells
Table 2
expressing b-ARs.
b-AR agonistic activity of thiourea derivatives
b
Values in parentheses represent the intrinsic activity (IA) given as a fraction of
the maximum stimulation with isoproterenol.
OH
H
O
N
S
R
N
N
H
H
showed a slight decrease in potency at b3-AR. These results indi-
cated that the introduction of any substituent at the 4-position
on the phenyl ring of the thiourea moiety in 14b may maintain po-
tency at b3-AR, but that intrinsic activity may tend to decrease.
Given the results of the in vitro study, compounds 18e, 18i, 18k,
and 18l were selected for in vivo evaluation in a rodent model of
type II diabetes (Table 4). The compounds were administered oral-
ly for 4 days in diabetic kk mice and the effects on plasma glucose
were measured. All compounds induced a significant reduction in
plasma glucose levels at the dose of 30 mg/kg. In particular, 18k
and 18l exhibited more potent hypoglycemic activities (46% and
40% decrease, respectively) than 2 (15% decrease) at the dose of
10 mg/kg.
M) (IAb)
a
Compound
R
EC50
(l
b3-AR
b2-AR
b1-AR
14b
0.44 (0.70)
>100 (0)
>100 (0.11)
18a
18b
18c
1.4 (0.46)
0.47 (0.53)
0.51 (0.34)
>100 (0.03)
>100 (0.03)
>100 (0.01)
>100 (0.05)
>100 (0.04)
>100 (0.03)
4. Conclusion
a
Agonistic activity was assessed by measuring cAMP accumulation in CHO cells
We have identified a new series of thiourea-based b3-AR
agonists and described their synthesis and SARs. Among these
compounds, the 4-nitrophenyl (18i), 3-methoxyphenyl (18k), and
4-methoxyphenyl (18l) derivatives showed potent agonistic activ-
expressing b-ARs.
b
Values in parentheses represent the intrinsic activity (IA) given as a fraction of
the maximum stimulation with isoproterenol.
ity at the b3-AR (EC50 = 0.10, 0.16, and 0.20
lM, respectively), and
also had functional selectivity over b1- and b2-ARs. In addition,
18k and 18l exhibited significant hypoglycemic activity in diabetic
kk mice at the dose of 10 mg/kg po.
moiety in 14b (18j–l) led to a several fold increase in the potency
of b3-AR relative to 14b, with a rank order of potency similar to
those of the chlorophenyl derivatives The introduction of a meta-
methoxy group (18k) led to a 2.8-fold increase in potency at the
b3-AR (EC50 = 0.16 lM) relative to 14b. These results revealed that
Table 4
the introduction of both electron-withdrawing and electron-
donating groups at the 3- or 4-position on the phenyl ring of the
thiourea moiety might be effective in improving agonistic activity
at the b3-AR while maintaining functional selectivity over b1- and
b2-ARs. Next, the introduction of another halogen group at the 4-
position on the phenyl ring of the thiourea moiety in 14b (18m–
o) yielded similar results to those of 4-chlorophenyl derivative
18f. Introduction of the methoxycarbonyl (18p) or sulfamoyl
(18q) group at the 4-position on the phenyl ring of the thiourea
moiety in 14b resulted in slightly increased potency at the b3-AR
relative to 14b; however, their intrinsic activities were decreased
(IA = 0.37 and 0.49, respectively). 4-Methylphenyl derivative 18r
Oral hypoglycemic activity in kk mice
Compound
Percent reduction in plasma glucosea
10 mg/kg
30 mg/kg
c
**
18e
18i
18k
18l
NTb
NT
29
32
49
53
**
**
***
46
**
***
40
*
**
2
15
38
a
The compounds were administered orally to male kk mice for 4 days.
Not tested.
b
c
*
**
***
Statistically significant at ( ) p < 0.05, ( ) p < 0.01, ( ) p < 0.001.

5514
T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
7.42–7.47 (3H, m), 7.56 (2H, d, J = 6.1 Hz), 8.13 (1H, s), 9.27 (1H,
5. Experimental
5.1. Chemistry
s); MS (FAB) m/z: 303 (MH+).
5.1.4. (S)-1-Phenoxy-3-(N-benzyl-{2-[4-(phenylamino)phenyl]-
ethyl}amino)-2-propanol (8)
Melting points were determined with a Yanaco MP-500D melt-
ing point apparatus and are uncorrected. 1H NMR spectra were re-
corded on a JEOL EX90, EX400, or GX500 spectrometer, and the
chemical shifts are expressed in d (ppm) values with tetramethyl-
silane as an internal standard (NMR description key: s = singlet,
d = doublet, t = triplet, m = multiplet, and br = broad peak). Mass
spectra were recorded on a Hitachi M-80 or JEOL JMS-DX300 spec-
trometer. The elemental analyses were performed with a Yanaco
MT-5 microanalyzer (C, H, N) and were within 0.4% of the theoret-
ical values. During the work-up, all organic solutions were dried
over anhydrous Mg2SO4.
A
mixture of compound 7 (0.27 g) and (S)-2-(phenoxy-
methyl)oxirane (0.31 g) in 2-propanol (15 ml) and dioxane (1 ml)
was refluxed for 24 h, and the resultant mixture was concentrated
in vacuo. The residue was purified using column chromatography
on silica gel with CHCl3/MeOH (50:1) as the eluent to yield 8
(0.36 g) as a colorless oil. 89% yield; 1H NMR (CDCl3) d: 2.69–2.88
(6H, m), 3.61 (1H, d, J = 14.0 Hz), 3.84–3.94 (3H, m), 3.98–4.05
(1H, m), 5.60 (1H, s), 6.83–7.03 (10H, m), 7.21–7.33 (9H, m); MS
(FAB) m/z: 453 (MH+).
5.1.5. (S)-1-Phenoxy-3-({2-[4-(phenylamino)phenyl]ethyl}amino)-
2-propanol hydrochloride (9)
5.1.1. (S)-N-Phenyl-4-{2-[(2-hydroxy-3-phenoxypropyl)amino]-
ethyl}benzenesulfonamide hydrochloride (4)
To a solution of 8 (0.34 g) in ethanol (15 ml) was added palla-
dium–carbon (10% w/w, 0.14 g), and the mixture was stirred under
a hydrogen atmosphere for 18 h. The catalyst was removed by fil-
tration, and the filtrate was concentrated in vacuo. To the solution
of the residue in methanol (10 ml) was added 1 M HCl–EtOAc
(0.7 ml), and the resultant mixture was concentrated in vacuo.
The residue was purified by recrystallization from ethanol–diethyl
ether to yield 9 (0.13 g) as a colorless solid. 43% yield; mp 160–
162 °C (EtOH–Et2O); 1H NMR (DMSO-d6) d: 2.85–3.07 (3H, m),
3.12–3.24 (3H, m), 3.94–4.02 (2H, m), 4.21–4.25 (1H, m), 5.91
(1H, d, J = 5.6 Hz), 6.78–6.82 (1H, m), 6.94–6.97 (3H, m), 7.02–
7.05 (4H, m), 7.20 (2H, d, J = 8.4 Hz), 7.19–7.23 (2H, m), 7.28–
7.33 (2H, m), 8.15 (1H, s), 8.84 (1H, br s), 9.04 (1H, br s); MS
(FAB) m/z: 363 (MH+). Anal. Calcd for C23H26N2O2ꢀ1.1HCl: C,
67.41; H, 6.86; N, 6.84; Cl, 9.52. Found: C, 67.52; H, 6.63; N,
6.77; Cl, 9.58.
A mixture of a commercially available N-phenyl-4-(2-amino-
ethyl)benzenesulfonamide (3) (0.13 g) and (S)-2-(phenoxy-
methyl)oxirane (0.07 g) in acetonitrile (5 ml) was refluxed for
24 h, and the resultant mixture was concentrated in vacuo. The
residue was purified using column chromatography on silica gel
with CHCl3/MeOH (5:1) as the eluent to yield free base (0.10 g).
To the solution of the residue in methanol (5 ml) was added 4 M
HCl–dioxane (0.1 ml), and the resultant mixture was concentrated
in vacuo. The residue was purified by recrystallization from meth-
anol-isopropanol to yield 4 (0.06 g) as a colorless solid. 28% yield;
mp 161–165 °C (MeOH–iPrOH); 1H NMR (DMSO-d6) d: 3.01–3.06
(3H, m), 3.15–3.25 (3H, m), 3.92–4.00 (2H, m), 4.19–4.21 (1H,
m), 5.89 (1H, d, J = 4.9 Hz), 6.93–7.03 (4H, m), 7.09–7.12 (2H, m),
7.20–7.24 (2H, m), 7.28–7.32 (2H, m), 7.44 (2H, d, J = 8.3 Hz),
7.73 (2H, d, J = 8.3 Hz), 8.88 (1H, br s), 9.06 (1H, br s), 10.31 (1H,
s); MS (FAB) m/z: 427 (MH+). Anal. Calcd for C23H26N2O4Sꢀ
HClꢀ0.5H2O: C, 58.53; H, 5.98; N, 5.94; S, 6.79; Cl, 7.51. Found: C,
58.49; H, 5.89; N, 5.91; S, 6.80; Cl, 7.67.
5.1.6. (S)-N-(4-{2-[N0-tert-Butoxycarbonyl-N0-(2-hydroxy-3-
phenoxypropyl)amino]ethyl}phenyl)benzamide (11a)
To a solution of tert-butyl (S)-N-[2-(4-aminophenyl)ethyl]-
N-(2-hydroxy-3-phenoxy)propylcarbamate (10) (0.48 g) in chloro-
form (15 ml) were added triethylamine (0.15 g) and benzoyl
chloride (0.19 g), and the mixture was stirred at room temperature
for 30 min. The resulting mixture was concentrated in vacuo. The
residue was purified using column chromatography on silica gel
with CHCl3/MeOH (30:1) as the eluent to yield 11a (0.48 g) as a col-
orless powder. 78% yield; 1H NMR (CDCl3) d: 1.47 (9H, s), 2.80–2.90
(2H, m), 3.28–3.50 (4H, m), 3.88–3.95 (2H, m), 4.11 (2H, br s),
6.89–6.98 (3H, m), 7.13–7.31 (4H, m), 7.47–7.57 (5H, m), 7.78–
7.87 (3H, m); MS (FAB) m/z: 491 (MH+).
5.1.2. N-Benzyl-4-(phenylamino)phenylacetamide (6)
To a solution of N-benzyl-4-aminophenylacetamide (5) (1.26 g)
in dichloromethane (70 ml) were added triphenylbismuth (3.86 g)
and copper diacetate (0.96 g) at room temperature, and the mix-
ture was refluxed for 3.5 h. After cooling to room temperature,
the solid was removed by filtration over Celite, and the filtrate
was concentrated in vacuo. The residue was purified using column
chromatography on silica gel with CHCl3/MeOH (40:1) as the elu-
ent to yield 6 (1.27 g) as a colorless powder. 76% yield; 1H NMR
(DMSO-d6) d: 3.38 (2H, s), 3.07–3.09 (2H, m), 4.27 (2H, d,
J = 6.1 Hz), 6.77–6.80 (1H, m), 7.00–7.04 (4H, m), 7.13–7.24 (7H,
m), 7.28–7.32 (2H, m), 8.06 (1H, s), 8.43–8.46 (1H, m); MS (FAB)
m/z: 317 (MH+).
5.1.7. Phenyl (S)-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-
phenoxypropyl)amino]ethyl}phenyl)carbamate (11b)
The title compound was prepared in the same manner as de-
scribed for 11a using phenyl chloroformate instead of benzoyl
chloride as a colorless powder. 77% yield; 1H NMR (CDCl3) d:
1.46 (9H, s), 2.75–2.90 (2H, m), 3.40–3.50 (4H, m), 3.89–3.95 (2H,
m), 4.09 (2H, br s), 6.89–6.98 (3H, m), 7.12–7.41 (11H, m); MS
(FAB) m/z: 507 (MH+).
5.1.3. N-Benzyl-2-[4-(phenylamino)phenyl]ethylamine (7)
To a solution of compound 6 (1.25 g) in tetrahydrofuran (10 ml)
was added 1 M borane–THF solution (12 ml), and the mixture was
refluxed for 1 h. After cooling to room temperature, to the resultant
mixture was added dropwise methanol (10 ml), and the mixture
was refluxed for 30 min. To the mixture was added concentrated
HCl aqueous solution (1.4 ml), and the mixture was heated at
80 °C for 2 h. After cooling to room temperature, the mixture was
diluted with 1 M sodium hydroxide solution (15 ml) and stirred.
The solid was isolated by filtration, and washed with water. The
residue was purified by recrystallization from ethanol–ethyl ace-
tate to yield 7 (0.79 g) as a colorless solid. 66% yield; 1H NMR
(DMSO-d6) d: 2.88–2.91 (2H, m), 3.07–3.09 (2H, m), 4.17 (2H, s),
6.80 (1H, t, J = 7.3 Hz), 7.02–7.11 (6H, m), 7.19–7.23 (2H, m),
5.1.8. (S)-N-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}
phenyl)benzamide hydrochloride (12a)
To a solution of 11a (0.47 g) in ethanol (10 ml) was added 4 M
HCl–EtOAc (10 ml), and the mixture was stirred at room tempera-
ture for 30 min. The resulting mixture was concentrated in vacuo.
The crude solid was purified by recrystallization from methanol to
yield 12a (0.20 g) as a colorless solid. 77% yield; mp 247–252 °C
(MeOH); 1H NMR (DMSO-d6) d: 2.94–3.08 (3H, m), 3.17–3.20

T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
5515
(3H, m), 3.94–4.02 (2H, m), 4.21–4.23 (1H, m), 5.90 (1H, d,
J = 4.8 Hz), 6.94–6.97 (3H, m), 7.23–7.33 (4H, m), 7.51–7.61 (3H,
m), 7.75 (2H, d, J = 8.4 Hz), 7.94–7.97 (2H, m), 8.81 (1H, br s),
8.94 (1H, br s), 10.26 (1H, s); MS (FAB) m/z: 391 (MH+). Anal. Calcd
for C24H26N2O3ꢀHClꢀ0.1H2O: C, 67.23; H, 6.39; N, 6.53; Cl, 8.27.
Found: C, 67.08; H, 6.34; N, 6.51; Cl, 8.08.
C
24H27N3O2SꢀHCl: C, 62.94; H, 6.16; N, 9.17; S, 7.00; Cl, 7.74. Found:
C, 62.87; H, 6.03; N, 9.19; S, 6.85; Cl, 7.63.
5.1.14. (S)-2-Cyano-1-(4-{2-[N-tert-butoxycarbonyl-N-(2-
hydroxy-3-phenoxypropyl)amino]ethyl}phenyl)-3-phenyl-
guanidine (15)
To a solution of 13b (0.40 g) in N,N-dimethylformamide (10 ml)
were added 1-ethyl-3-(30-dimethylaminopropyl)carbodiimide
hydrochloride (0.16 g), cyanamide (0.06 g) and diisopropylethyl-
amine (0.01 g), and the mixture was heated at 50 °C for 20 h. The
resulting mixture was concentrated in vacuo, 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 (1:1) as the eluent to yield 15 (0.15 g) as a colorless
oil. 37% yield; 1H NMR (CDCl3) d: 1.46 (9H, s), 2.80–2.90 (2H, m),
3.35–3.56 (4H, m), 3.82–4.10 (3H, m), 6.85–7.00 (4H, m), 7.20–
7.44 (10H, m); MS (FAB) m/z: 530 (MH+).
5.1.9. Phenyl (S)-(4-{2-[(2-hydroxy-3-phenoxypropyl)amino]
ethyl}phenyl)carbamate hydrochloride (12b)
The title compound was prepared in the same manner as de-
scribed for 12a using 11b instead of 11a as a colorless solid. 54%
yield; mp 215–219 °C (MeOH–EtOH); 1H NMR (DMSO-d6) d:
2.93–3.07 (3H, m), 3.15–3.22 (3H, m), 3.94–4.01 (2H, m), 4.21–
4.24 (1H, m), 5.90 (1H, d, J = 4.8 Hz), 6.94–6.97 (3H, m), 7.20–
7.33 (7H, m), 7.41–7.48 (4H, m), 8.85 (1H, br s), 9.03 (1H, br s),
10.22 (1H, s); MS (FAB) m/z: 407 (MH+). Anal. Calcd for
C24H26N2O4ꢀHCl: C, 65.08; H, 6.14; N, 6.32; Cl, 8.00. Found: C,
65.14; H, 5.90; N, 6.38; Cl, 7.89.
5.1.10. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-
phenoxypropyl)amino]ethyl}phenyl)-3-phenylurea (13a)
To a solution of 10 (0.49 g) in chloroform (15 ml) was added
phenylisocyanate (0.17 g), and the mixture was stirred at 60 °C
for 3 h. The resulting mixture was concentrated in vacuo. The res-
idue was purified using column chromatography on silica gel with
CHCl3/MeOH (30:1) as the eluent to yield 13a (0.36 g) as a colorless
powder. 56% yield; 1H NMR (CDCl3) d: 1.46 (9H, s), 2.80–2.90 (2H,
m), 3.42–3.50 (4H, m), 3.88–3.95 (2H, m), 4.06–4.11 (2H, m), 6.60–
6.67 (2H, m), 6.87–7.00 (3H, m), 7.05–7.12 (4H, m), 7.26–7.52 (7H,
m); MS (FAB) m/z: 506 (MH+).
5.1.15. (S)-2-Cyano-1-(4-{2-[(2-hydroxy-3-phenoxypropyl)-
amino]ethyl}phenyl)-3-phenylguanidine hydrochloride (16)
A mixture of 15 (0.15 g) in trifluoroacetic acid (5 ml) was stirred
at room temperature for 5 min, and the resultant mixture was con-
centrated in vacuo. To the residue was added 4 M HCl–dioxane
(5 ml), the mixture was stirred at room temperature for 30 min,
and the resultant mixture was concentrated in vacuo. The residue
was purified by recrystallization from aqueous methanol to yield
16 (0.05 g) as a colorless solid. 37% yield; mp 190–193 °C (MeOH–
water); 1H NMR (DMSO-d6) d: 2.90–3.11 (3H, m), 3.12–3.30 (3H,
m), 3.93–4.02 (2H, m), 4.21 (1H, br s), 5.90 (1H, d, J = 4.9 Hz), 6.93–
6.99 (3H, m), 7.10–7.37 (11H, m), 8.78 (1H, br s), 8.94 (1H, br s),
9.50 (2H, br s); MS (FAB-nega.) m/z: 428 [(MꢁH)ꢁ]. Anal. Calcd for
C25H27N5O2ꢀHClꢀ0.3H2O: C, 63.70; H, 6.12; N, 14.86; Cl, 7.52. Found:
C, 63.92; H, 6.04; N, 14.45; Cl, 7.54.
5.1.11. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-
phenoxypropyl)amino]ethyl}phenyl)-3-phenylthiourea (13b)
To a solution of 10 (0.50 g) in chloroform (15 ml) was added
phenylisothiocyanate (0.30 g), and the mixture was stirred at
80 °C for 15 h. The resulting mixture was concentrated in vacuo.
The residue was purified using column chromatography on silica
gel with CHCl3/MeOH (30:1) as the eluent to yield 13b (0.35 g) as
a colorless powder. 52% yield; 1H NMR (CDCl3) d: 1.46 (9H, s),
2.80–2.95 (2H, m), 3.36–3.55 (4H, m), 3.85–4.05 (3H, m), 4.05–
4.15 (1H, m), 6.89 (2H, d, J = 8.0 Hz), 6.96 (1H, t, J = 7.2 Hz), 7.15–
7.25 (2H, m), 7.26–7.45 (9H, m), 7.68 (1H, s), 7.73 (1H, s); MS
(FAB) m/z: 522 (MH+).
5.1.16. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-
phenoxypropyl)amino]ethyl}phenyl)-3-cyclohexylthiourea (17a)
The title compound was prepared in the same manner as de-
scribed for 13b using cyclohexylisothiocyanate instead of phenyli-
sothiocyanate as a colorless oil. 20% yield; 1H NMR (CDCl3) d:
1.05–1.20 (2H, m), 1.45 (9H, s), 1.30–1.70 (6H, m), 2.00–2.10 (2H,
m), 2.70–3.00 (3H, m), 3.25–3.60 (4H, m), 3.80–4.00 (2H, m),
4.05–4.15 (1H, m), 4.20–4.35 (1H, m), 5.83 (1H, d, J = 8.0 Hz),
6.90 (2H, d, J = 7.6 Hz), 6.98 (1H, t, J = 7.2 Hz), 7.00 (2H, d,
J = 8.0 Hz), 7.15–7.32 (5H, m), 7.39 (1H, s); MS (FAB) m/z: 528
(MH+).
5.1.12. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}
phenyl)-3-phenylurea hydrochloride (14a)
The title compound was prepared in the same manner as de-
scribed for 12a using 13a instead of 11a as a colorless solid. 68%
yield; mp 217–219 °C (MeOH); 1H NMR (DMSO-d6) d: 2.86–2.98
(2H, m), 3.03–3.06 (1H, m), 3.16–3.20 (3H, m), 3.93–4.01 (2H, m),
4.20 (1H, br s), 5.90 (1H, br s), 6.93–6.97 (4H, m), 7.17 (2H, d,
J = 8.8 Hz), 7.24–7.33 (4H, m), 7.41–7.46 (4H, m), 8.73 (1H, br s),
8.84 (1H, br s), 9.05 (1H, d, J = 5.2 Hz); MS (FAB) m/z: 406 (MH+).
Anal. Calcd for C24H27N3O3ꢀHCl: C, 65.22; H, 6.39; N, 9.51; Cl,
8.02. Found: C, 65.12; H, 6.27; N, 9.53; Cl, 7.83.
5.1.17. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-
phenoxypropyl)amino]ethyl}phenyl)-3-benzylthiourea (17b)
The title compound was prepared in the same manner as de-
scribed for 13b using benzylisothiocyanate instead of phenylisothi-
ocyanate as a colorless oil. 55% yield; 1H NMR (CDCl3) d: 1.42 (9H, s),
2.75–2.93 (2H, m), 3.40–3.50 (4H, m), 3.49 (2H, d, J = 7.6 Hz), 3.80–
4.00 (2H, m), 4.00–4.10 (1H, m), 4.87 (1H, d, J = 7.2 Hz), 6.23 (1H,
br s), 6.88 (2H, d, J = 8.4 Hz), 6.97 (1H, t, J = 7.2 Hz), 7.08–7.44
(13H, m), 7.64 (1H, s); MS (FAB) m/z: 536 (MH+).
5.1.13. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-phenylthiourea hydrochloride (14b)
5.1.18. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(2-phenylethyl)thiourea (17c)
The title compound was prepared in the same manner as de-
scribed for 13b using 2-phenylethylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 59% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.86–2.94 (4H, m), 3.34–3.54 (4H, m),
3.82–4.02 (5H, m), 4.04–4.12 (1H, m), 5.94 (1H, br s), 6.90 (4H, d,
J = 8.4 Hz), 6.98 (1H, t, J = 7.2 Hz), 7.05–7.34 (9H, m), 7.54 (1H, s);
MS (FAB) m/z: 550 (MH+).
The title compound was prepared in the same manner as de-
scribed for 12a using 13b instead of 11a as a colorless solid. 57%
yield; mp 214–217 °C (MeOH–EtOH); 1H NMR (DMSO-d6) d:
2.93–3.07 (3H, m), 3.15–3.25 (3H, m), 3.94–4.01 (2H, m), 4.19–
4.23 (1H, m), 5.91 (1H, d, J = 4.4 Hz), 6.95–6.97 (3H, m), 7.09–
7.13 (1H, m), 7.22 (2H, d, J = 8.0 Hz), 7.29–7.34 (4H, m),
7.50–7.55 (4H, m), 8.79 (1H, br s), 8.93 (1H, br s), 10.17 (1H, s),
10.18 (1H, s); MS (FAB) m/z: 422 (MH+). Anal. Calcd for

5516
T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
5.1.19. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(2-chlorophenyl)thiourea
(17d)
5.1.25. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(2-methoxyphenyl)thiourea
(17j)
The title compound was prepared in the same manner as de-
scribed for 13b using 2-chlorophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 34% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.75–2.95 (2H, m), 3.25–3.55 (4H, m),
3.80–4.00 (3H, m), 4.05–4.16 (1H, m), 6.89 (2H, d, J = 8.4 Hz),
6.95–7.00 (1H, m), 7.15–7.40 (10H, m), 7.71 (1H, s), 7.75 (1H, s);
MS (FAB) m/z: 556 (MH+).
The title compound was prepared in the same manner as de-
scribed for 13b using 2-methoxyphenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 54% yield; 1H NMR
(CDCl3) d: 1.37 (9H, s), 2.49–2.51 (2H, m), 3.04–3.17 (1H, m),
3.38–3.44 (3H, m), 3.83–3.85 (5H, m), 3.98 (1H, br s), 5.14–5.19
(1H, m), 6.90–6.94 (4H, m), 7.04–7.06 (1H, m), 7.12–7.16 (3H,
m), 7.26–7.30 (2H, m), 7.43 (2H, d, J = 8.0 Hz), 7.95 (1H, d,
J = 7.6 Hz), 9.07 (1H, s), 9.89 (1H, s); MS (FAB) m/z: 552 (MH+).
5.1.20. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(3-chlorophenyl)thiourea
(17e)
The title compound was prepared in the same manner as de-
scribed for 13b using 3-chlorophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 69% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.80–2.95 (2H, m), 3.40–3.60 (4H, m),
3.80–4.00 (3H, m), 4.00–4.10 (1H, m), 6.88 (2H, d, J = 8.0 Hz),
6.97 (1H, t, J = 7.2 Hz), 7.20–7.40 (9H, m), 7.44 (1H, s), 7.60 (1H,
s), 7.71 (1H, s); MS (FAB) m/z: 556 (MH+).
5.1.26. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(3-methoxyphenyl)thiourea
(17k)
The title compound was prepared in the same manner as de-
scribed for 13b using 3-methoxyphenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 28% yield; 1H NMR
(CDCl3) d: 1.38 (9H, s), 2.74–2.79 (2H, m), 3.05–3.17 (1H, m),
3.38–3.43 (3H, m), 3.73 (3H, s), 3.83–3.88 (2H, m), 3.97–4.00
(1H, m), 5.14–5.19 (1H, m), 6.69 (1H, dd, J = 8.0, 2.0 Hz), 6.90–
6.94 (3H, m), 7.01 (1H, dd, J = 8.0, 1.6 Hz), 7.12–7.14 (2H, m),
7.18–7.30 (4H, m), 7.38–7.40 (2H, m), 9.72 (2H, s); MS (FAB) m/z:
552 (MH+).
5.1.21. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-chlorophenyl)thiourea
(17f)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-chlorophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 51% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.84–2.94 (2H, m), 3.37–3.54 (4H, m),
3.70–4.00 (3H, m), 4.00–4.20 (1H, m), 6.88 (2H, d, J = 8.0 Hz),
6.97 (1H, t, J = 7.2 Hz), 7.55–7.70 (10H, m), 7.56 (1H, s), 7.65 (1H,
s); MS (FAB) m/z: 556 (MH+).
5.1.27. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-methoxyphenyl)thiourea
(17l)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-methoxyphenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 53% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.80–2.95 (2H, m), 3.35–3.55 (4H, m),
3.82 (3H, s), 3.80–4.00 (3H, m), 4.05–4.15 (1H, m), 6.86–7.00 (6H,
m), 7.13–7.20 (2H, m), 7.24–7.34 (5H, m), 7.51 (1H, s), 7.57 (1H,
s); MS (FAB) m/z: 552 (MH+).
5.1.22. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(2-nitrophenyl)thiourea (17g)
The title compound was prepared in the same manner as de-
scribed for 13b using 2-nitrophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 85% yield; 1H NMR
(CDCl3) d: 1.38 (9H, s), 2.77–2.81 (2H, m), 3.10–3.18 (1H, m),
3.40–3.48 (3H, m), 3.82–3.88 (2H, m), 3.89–4.00 (1H, m), 5.15–
5.18 (1H, m), 6.90–6.94 (3H, m), 7.18–7.19 (2H, m), 7.26–7.31
(2H, m), 7.39–7.43 (3H, m), 7.67–7.76 (2H, m), 8.00 (1H, dd,
J = 8.4, 1.6 Hz), 9.84 (1H, s), 10.34 (1H, s); MS (FAB) m/z: 567
(MH+).
5.1.28. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-fluorophenyl)thiourea
(17m)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-fluorophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 34% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.72–2.80 (2H, m), 3.34–3.46 (4H, m),
3.80–3.90 (2H, m), 3.93–4.03 (1H, m), 5.12–5.21 (1H, m), 6.90–
6.95 (3H, m), 7.10–7.19 (4H, m), 7.26–7.30 (2H, m), 7.35–7.40
(2H, m), 7.43–7.48 (2H, m), 9.66 (1H, s), 9.72 (1H, s); MS (FAB)
m/z: 540 (MH+).
5.1.23. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(3-nitrophenyl)thiourea (17h)
The title compound was prepared in the same manner as de-
scribed for 13b using 3-nitrophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 62% yield; 1H NMR
(CDCl3) d: 1.38 (9H, s), 2.77–2.80 (2H, m), 3.06–3.17 (1H, m),
3.37–3.44 (3H, m), 3.82–3.87 (2H, m), 3.99 (1H, br s), 5.15–5.18
(1H, m), 6.91–6.94 (3H, m), 7.16–7.19 (2H, m), 7.28 (2H, t,
J = 8.4 Hz), 7.90 (1H, dd, J = 8.0, 2.0 Hz), 7.95 (1H, dd, J = 8.0,
2.0 Hz), 8.56 (1H, t, J = 2.0 Hz), 10.05 (1H, s), 10.08 (1H, s); MS
(FAB) m/z: 567 (MH+).
5.1.29. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-bromophenyl)thiourea
(17n)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-bromophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 37% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.80–2.95 (2H, m), 3.36–3.57 (4H, m),
3.80–4.00 (2H, m), 4.00–4.08 (1H, m), 6.88 (2H, d, J = 8.4 Hz),
6.97 (1H, t, J = 7.2 Hz), 7.20–7.32 (8H, m), 7.48 (2H, d, J = 8.8 Hz),
7.57 (1H, s), 7.69 (1H, s); MS (FAB) m/z: 600 (MH+).
5.1.24. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-nitrophenyl)thiourea (17i)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-nitrophenylisothiocyanate instead of phen-
ylisothiocyanate as a colorless powder. 88% yield; 1H NMR (CDCl3)
d: 1.47 (9H, s), 2.85–3.00 (3H, m), 3.35–3.62 (3H, m), 3.80–4.05
(3H, m), 6.86 (2H, d, J = 7.6 Hz), 6.97 (1H, t, J = 7.2 Hz), 7.20–7.32
(6H, m), 7.71 (2H, d, J = 8.8 Hz), 7.84 (1H, s), 7.99 (1H, s), 8.16
(2H, d, J = 8.8 Hz); MS (FAB) m/z: 567 (MH+).
5.1.30. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-iodophenyl)thiourea (17o)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-iodophenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 43% yield; 1H NMR

T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
5517
(CDCl3) d: 1.46 (9H, s), 2.80–2.95 (2H, m), 3.35–3.57 (4H, m),
3.80–4.00 (3H, m), 4.00–4.10 (1H, m), 6.88 (2H, d, J = 8.0 Hz),
6.97 (1H, t, J = 7.2 Hz), 7.17 (2H, d, J = 8.4 Hz), 7.20–7.32 (6H,
m), 7.58 (1H, s), 7.67 (2H, d, J = 8.4 Hz), 7.72 (1H, s); MS (FAB)
m/z: 648 (MH+).
5.1.36. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(2-phenylethyl)thiourea hydrochloride (18c)
The title compound was prepared in the same manner as de-
scribed for 12a using 17c instead of 11a as a colorless solid. 64%
yield; mp 183–185 °C (MeOH–EtOH); 1H NMR (DMSO-d6) d:
2.84–3.08 (5H, m), 3.16–3.23 (3H, m), 3.65–3.73 (2H, m), 3.94–
4.01 (2H, m), 4.17–4.25 (1H, m), 5.90 (1H, d, J = 5.2 Hz), 6.94–
6.98 (3H, m), 7.17–7.36 (11H, m), 7.87 (1H, br s), 8.79 (2H, br s),
9.73 (1H, br s); MS (FAB) m/z: 450 (MH+). Anal. Calcd for
C26H31N3O2SClꢀHCl: C, 64.25; H, 6.64; N, 8.64; S, 6.60; Cl, 7.29.
Found: C, 64.11; H, 6.80; N, 8.56; S, 6.52; Cl, 7.42.
5.1.31. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-methoxycarbonylphenyl)-
thiourea (17p)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-methoxycarbonylphenylisothiocyanate in-
stead of phenylisothiocyanate as a colorless powder. 56% yield; 1H
NMR (CDCl3) d: 1.46 (9H, s), 2.80–2.96 (2H, m), 3.36–3.60 (4H, m),
3.84–4.00 (2H, m), 3.91 (3H, s), 4.00–4.04 (1H, m), 6.88 (2H, d,
J = 8.0 Hz), 6.97 (1H, t, J = 7.2 Hz), 7.22–7.32 (6H, m), 7.53 (2H, d,
J = 8.4 Hz), 7.71 (1H, s), 7.77 (1H, s), 8.03 (2H, d, 6.88 (2H, d,
J = 8.0 Hz); MS (FAB) m/z: 580 (MH+).
5.1.37. (S)-1-(2-Chlorophenyl)-3-(4-{2-[(2-hydroxy-3-phenoxy-
propyl)amino]ethyl}phenyl)thiourea hydrochloride (18d)
The title compound was prepared in the same manner as de-
scribed for 12a using 17d instead of 11a as a colorless solid. 43%
yield; mp 157–160 °C (EtOH); 1H NMR (DMSO-d6) d: 2.90–3.10
(3H, m), 3.15–3.26 (3H, m), 3.94–4.01 (2H, m), 4.16–4.25 (1H,
m), 5.90 (1H, br s), 6.94–6.98 (3H, m), 7.20–7.36 (6H, m), 7.49–
7.55 (3H, m), 7.61–7.64 (1H, m), 8.78 (1H, br s), 8.91 (1H, br s),
9.60 (1H, s), 10.30 (1H, s); MS (FAB) m/z: 456 (MH+). Anal. Calcd
for C24H26N3O2SClꢀHCl: C, 58.58; H, 5.53; N, 8.53; S, 6.51; Cl,
14.40. Found: C, 58.36; H, 5.52; N, 8.44; S, 6.27; Cl, 14.44.
5.1.32. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-sulfamoylphenyl)thiourea
(17q)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-sulfamoylphenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 61% yield; 1H NMR
(CDCl3) d: 1.46 (9H, s), 2.75–2.90 (2H, m), 3.20–3.55 (4H, m),
3.88–3.97 (2H, m), 4.05–4.15 (1H, m), 6.47 (2H, s), 6.88–6.97
(3H, m), 7.12–7.44 (6H, m), 7.73 (2H, d, J = 8.0 Hz), 7.87 (2H, d,
J = 8.8 Hz), 9.32 (2H, br s); MS (FAB) m/z: 601 (MH+).
5.1.38. (S)-1-(3-Chlorophenyl)-3-(4-{2-[(2-hydroxy-3-phenoxy-
propyl)amino]ethyl}phenyl)thiourea hydrochloride (18e)
The title compound was prepared in the same manner as de-
scribed for 12a using 17e instead of 11a as a colorless solid. 51%
yield; mp 154–156 °C (EtOH); 1H NMR (DMSO-d6) d: 2.91–3.09
(3H, m), 3.15–3.25 (3H, m), 3.94–4.01 (2H, m), 4.18–4.26 (1H,
m), 5.91 (1H, br s), 6.94–6.97 (3H, m), 7.14–7.17 (1H, m), 7.23
(2H, d, J = 8.4 Hz), 7.29–7.36 (3H, m), 7.44–7.54 (3H, m), 7.82
(1H, t, J = 2.0 Hz), 8.79 (1H, br s), 8.93 (1H, br s), 10.38 (1H, s),
10.44 (1H, s); MS (FAB) m/z: 456 (MH+). Anal. Calcd for
C24H26N3O2SClꢀHCl: C, 58.58; H, 5.53; N, 8.53; S, 6.51; Cl, 14.40.
Found: C, 58.26; H, 5.60; N, 8.43; S, 6.30; Cl, 14.47.
5.1.33. (S)-1-(4-{2-[N-tert-Butoxycarbonyl-N-(2-hydroxy-3-phen-
oxypropyl)amino]ethyl}phenyl)-3-(4-methylphenyl) thiourea
(17r)
The title compound was prepared in the same manner as de-
scribed for 13b using 4-methylphenylisothiocyanate instead of
phenylisothiocyanate as a colorless powder. 41% yield; 1H NMR
(CDCl3) d: 1.38 (9H, s), 2.28 (3H, s), 2.74–2.78 (2H, m), 3.05–
3.17 (1H, m), 3.36–3.44 (3H, m), 3.82–3.87 (2H, m), 3.98 (1H, br
s), 5.13–5.17 (1H, m), 6.90–6.94 (3H, m), 7.12–7.14 (4H, m),
7.26–7.40 (6H, m), 9.60 (1H, s), 9.61 (1H, s); MS (FAB) m/z: 536
(MH+).
5.1.39. (S)-1-(4-Chlorophenyl)-3-(4-{2-[(2-hydroxy-3-phenoxy-
propyl)amino]ethyl}phenyl)thiourea hydrochloride (18f)
The title compound was prepared in the same manner as de-
scribed for 12a using 17f instead of 11a as a colorless solid. 57%
yield; mp 211–214 °C (MeOH–EtOH); 1H NMR (DMSO-d6) d:
2.90–3.08 (3H, m), 3.15–3.25 (3H, m), 3.94–4.01 (2H, m), 4.16–
4.25 (1H, m), 5.90 (1H, d, J = 4.8 Hz), 6.94–6.98 (3H, m), 7.22 (2H,
d, J = 8.4 Hz), 7.29–7.39 (4H, m), 7.49 (2H, d, J = 8.4 Hz), 7.58 (2H,
d, J = 8.4 Hz), 8.78 (1H, br s), 8.89 (1H, br s), 10.26 (1H, s), 10.29
(1H, s); MS (FAB) m/z: 456 (MH+). Anal. Calcd for
C24H26N3O2SClꢀHCl: C, 58.58; H, 5.53; N, 8.53; S, 6.51; Cl, 14.40.
Found: C, 58.24; H, 5.59; N, 8.42; S, 6.41; Cl, 14.40.
5.1.34. (S)-1-Cyclohexyl-3-(4-{2-[(2-hydroxy-3-phenoxypropyl)-
amino]ethyl}phenyl)thiourea hydrochloride (18a)
The title compound was prepared in the same manner as de-
scribed for 12a using 17a instead of 11a as a colorless solid. 65%
yield; mp 155–160 °C (EtOH–Et2O); 1H NMR (DMSO-d6) d: 1.10–
1.36 (5H, m), 1.52–1.60 (1H, m), 1.64–1.72 (2H, m), 1.84–1.95
(2H, m), 2.86–3.28 (7H, m), 3.97 (2H, t, J = 4.4 Hz), 4.15–4.25 (1H,
m), 5.88 (1H, br s), 6.93–7.00 (3H, m), 7.18 (2H, d, J = 8.4 Hz),
7.31 (2H, t, J = 8.0 Hz), 7.44 (2H, d, J = 8.4 Hz), 8.68 (2H, br s),
9.45 (1H, s); MS (FAB) m/z: 428 (MH+). Anal. Calcd for
C24H33N3O2Sꢀ1.3HClꢀ0.4H2O: C, 59.78; H, 7.34; N, 8.71; S, 6.65; Cl,
9.56. Found: C, 59.88; H, 7.37; N, 8.36; S, 5.81; Cl, 9.45.
5.1.40. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(2-nitrophenyl)thiourea hydrochloride (18g)
The title compound was prepared in the same manner as de-
scribed for 12a using 17g instead of 11a as a colorless powder.
89% yield; 1H NMR (DMSO-d6) d: 2.94–3.09 (3H, m), 3.16–3.24
(3H, m), 3.94–4.01 (2H, m), 4.20–4.25 (1H, m), 5.91 (1H, br s),
6.94–6.98 (3H, m), 7.25–7.33 (4H, m), 7.39–7.46 (1H, m), 7.50
(2H, d, J = 8.4 Hz), 7.70 (2H, d, J = 4.0 Hz), 8.01 (1H, d, J = 8.0 Hz),
10.09 (1H, s), 10.60 (1H, s); MS (FAB) m/z: 467 (MH+). Anal. Calcd
for C24H26N4O4SꢀHClꢀ0.2H2O: C, 56.90; H, 5.45; N, 11.06; S, 6.33;
Cl, 7.00. Found: C, 56.63; H, 5.33; N, 11.03; S, 6.39; Cl, 7.31.
5.1.35. (S)-1-Benzyl-3-(4-{2-[(2-hydroxy-3-phenoxypropyl)amino]-
ethyl}phenyl)thiourea hydrochloride (18b)
The title compound was prepared in the same manner as de-
scribed for 12a using 17b instead of 11a as a colorless solid. 43%
yield; mp 185–187 °C (EtOH-Et2O); 1H NMR (DMSO-d6) d: 2.88–
3.10 (3H, m), 3.13–3.27 (3H, m), 3.93–4.02 (2H, m), 4.15–4.24
(1H, m), 4.73 (2H, d, J = 6.0 Hz), 5.89 (1H, d, J = 5.2 Hz), 6.94–6.98
(3H, m), 7.21 (2H, d, J = 8.4 Hz), 7.23–7.35 (7H, m), 7.44 (2H, d,
J = 8.4 Hz), 8.28 (1H, br s), 8.73 (1H, br s), 8.84 (1H, br s), 9.79
(1H, br s); MS (FAB) m/z: 436 (MH+). Anal. Calcd for
C25H29N3O2SꢀHClꢀ0.2H2O: C, 63.13; H, 6.44; N, 8.83; S, 6.74; Cl,
7.45. Found: C, 63.18; H, 6.35; N, 8.82; S, 6.78; Cl, 7.43.
5.1.41. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(3-nitrophenyl)thiourea hydrochloride (18h)
Thetitlecompoundwaspreparedinthesamemannerasdescribed
for 12a using 17h instead of 11a as a colorless powder. 34% yield; 1H

5518
T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
NMR (DMSO-d6) d: 2.96–3.08 (3H, m), 3.19–3.24 (3H, m), 3.94–4.01
(2H, m), 4.22 (1H, br s), 5.90 (1H, br s), 6.95–6.97 (3H, m), 7.25–7.33
(4H, m), 7.50–7.52 (2H, m), 7.59–7.63 (1H, m), 7.92–7.96 (2H, m),
8.70(1H,s), 10.52(1H,s), 10.70(1H, s);MS(FAB)m/z:467(MH+).Anal.
Calcd for C24H26N4O4SꢀHClꢀ0.4H2O:C, 56.50; H, 5.49;N, 10.98; S, 6.28;
Cl, 6.95. Found: C, 56.43; H, 5.38; N, 10.91; S, 6.20; Cl, 7.17.
7.20–7.24 (2H, m), 7.31 (2H, t, J = 7.2 Hz), 7.47–7.54 (4H, m), 8.74
(2H, br s), 10.04 (2H, br s); MS (FAB) m/z: 440 (MH+). Anal. Calcd
for C24H26N3O2SFClꢀ1.1HCl: C, 60.10; H, 5.69; N, 8.76; S, 6.69; F,
3.96; Cl, 8.13. Found: C, 60.03; H, 5.67; N, 8.86; S, 6.44; F, 3.58;
Cl, 7.83.
5.1.47. (S)-1-(4-Bromophenyl)-3-(4-{2-[(2-hydroxy-3-phenoxy-
propyl)amino]ethyl}phenyl)thiourea hydrochloride (18n)
The title compound was prepared in the same manner as de-
scribed for 12a using 17n instead of 11a as a colorless solid. 63%
yield; mp 179–182 °C (MeOH–EtOH); 1H NMR (DMSO-d6) d:
2.90–3.10 (3H, m), 3.14–3.28 (3H, m), 3.94–4.02 (2H, m), 4.16–
4.25 (1H, m), 5.90 (1H, br s), 6.94–6.98 (3H, m), 7.23 (2H, d,
J = 8.0 Hz), 7.31 (2H, t, J = 8.0 Hz), 7.48–7.56 (6H, m), 8.78 (1H, br
s), 8.92 (1H, br s), 10.28 (1H, s), 10.31 (1H, s); MS (FAB) m/z: 500,
502 (MH+). Anal. Calcd for C24H26N3O2SBrꢀHClꢀ0.1H2O: C, 53.51;
H, 5.09; N, 7.80; S, 5.95; Br, 14.83; Cl, 6.58. Found: C, 53.57; H,
5.05; N, 7.79; S, 5.89; Br, 14.46; Cl, 6.62.
5.1.42. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(4-nitrophenyl)thiourea hydrochloride (18i)
The title compound was prepared in the same manner as de-
scribed for 12a using 17i insteadof 11a as a colorlesssolid. 55%yield;
mp 152–154 °C (EtOH); 1H NMR (DMSO-d6) d: 2.91–3.10 (3H, m),
3.16–3.28 (3H, m), 3.94–4.02 (2H, m), 4.17–4.25 (1H, m), 5.90 (1H,
br s), 6.94–6.98 (3H, m), 7.24–7.34 (4H, m), 7.53 (2H, d, J = 8.8 Hz),
7.95 (2H, d, J = 9.2 Hz), 8.21 (2H, d, J = 9.2 Hz), 8.76 (1H, br s), 10.69
(1H, s), 10.95 (1H, s); MS (FAB) m/z: 467 (MH+). Anal. Calcd for
C24H26N4O4SꢀHClꢀ0.2H2O: C, 56.90; H, 5.45; N, 11.06; S, 6.33; Cl,
7.00. Found: C, 56.95; H, 5.42; N, 10.92; S, 6.14; Cl, 7.17.
5.1.43. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(2-methoxyphenyl)thiourea hydrochloride (18j)
The title compound was prepared in the same manner as de-
scribed for 12a using 17j instead of 11a as a colorless powder.
67% yield; 1H NMR (DMSO-d6) d: 2.94–3.09 (3H, m), 3.16–3.24
(3H, m), 3.83 (3H, s), 3.96–4.00 (2H, m), 4.20–4.24 (1H, m), 5.91
(1H, br s), 6.90–6.98 (5H, m), 7.05–7.07 (1H, m), 7.13–7.17 (1H,
m), 7.22 (2H, d, J = 8.0 Hz), 7.29–7.34 (2H, m), 7.53 (2H, d,
J = 8.0 Hz), 7.91 (1H, d, J = 7.6, 1.2 Hz), 9.26 (1H, s), 10.18 (1H, s);
MS (FAB) m/z: 452 (MH+). Anal. Calcd for C25H29N3O3SꢀHClꢀ1.2H2O:
C, 58.92; H, 6.41; N, 8.24; S, 6.29; Cl, 6.96. Found: C, 59.11; H, 6.17;
N, 7.98; S, 6.09; Cl, 7.17.
5.1.48. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(4-iodophenyl)thiourea hydrochloride (18o)
The title compound was prepared in the same manner as de-
scribed for 12a using 17o instead of 11a as a colorless solid.
55% yield; mp 181–184 °C (MeOH); 1H NMR (DMSO-d6) d: 2.89–
3.06 (3H, m), 3.15–3.25 (3H, m), 3.93–4.01 (2H, m), 4.18–4.22
(1H, m), 5.90 (1H, d, J = 5.2 Hz), 6.94–6.98 (3H, m), 7.22 (2H, d,
J = 8.0 Hz), 7.29–7.33 (2H, m), 7.37–7.41 (2H, m), 7.48 (2H, d,
J = 8.0 Hz), 7.63–7.67 (2H, m), 8.75 (1H, br s), 8.85 (1H, br s),
10.20 (1H, s), 10.21 (1H, s); MS (FAB) m/z: 548 (MH+). Anal. Calcd
for C24H26N3O2SIꢀHCl: C, 49.37; H, 4.66; N, 7.20; S, 5.49; I, 21.73;
Cl, 6.07. Found: C, 49.46; H, 4.56; N, 7.16; S, 5.48; I, 21.83; Cl,
6.28.
5.1.44. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(3-methoxyphenyl)thiourea hydrochloride (18k)
The title compound was prepared in the same manner as de-
scribed for 12a using 17k instead of 11a as a colorless solid. 50%
yield; mp 151–152 °C (MeOH–EtOAc); 1H NMR (DMSO-d6) d:
2.90–3.09 (3H, m), 3.17–3.23 (3H, m), 3.73 (3H, s), 3.94–4.01 (2H,
m), 4.18–4.25 (1H, m), 5.90 (1H, d, J = 4.8 Hz), 6.69 (1H, dd,
J = 8.0, 2.0 Hz), 6.94–6.98 (3H, m), 7.05–7.08 (1H, m), 7.20–7.24
(3H, m), 7.28–7.33 (3H, m), 7.50 (2H, d, J = 8.4 Hz), 10.17 (1H, s),
10.18 (1H, s); MS (FAB) m/z: 452 (MH+). Anal. Calcd for
C25H29N3O3SꢀHCl: C, 61.53; H, 6.20; N, 8.61; S, 6.57; Cl, 7.26.
Found: C, 61.16; H, 6.25; N, 8.52; S, 6.47; Cl, 7.25.
5.1.49. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(4-methoxycarbonylphenyl)thiourea hydrochloride
(18p)
The title compound was prepared in the same manner as de-
scribed for 12a using 17p instead of 11a as a colorless solid. 40%
yield; mp 178–181 °C (MeOH–EtOAc); 1H NMR (DMSO-d6) d:
2.90–3.10 (3H, m), 3.12–3.28 (3H, m), 3.83 (3H, s), 3.94–4.02 (2H,
m), 4.16–4.26 (1H, m), 5.90 (1H, d, J = 4.4 Hz), 6.94–6.98 (3H, m),
7.24 (2H, d, J = 8.0 Hz), 7.31 (2H, t, J = 8.0 Hz), 7.52 (2H, d,
J = 8.0 Hz), 7.80 (2H, dd, J = 8.8, 2.8 Hz), 7.91 (2H, d, J = 8.8 Hz),
8.78 (1H, br s), 8.91 (1H, br s), 10.51 (1H, s), 10.65 (1H, s); MS
(FAB) m/z: 480 (MH+). Anal. Calcd for C26H29N3O4Sꢀ1.05HCl: C,
60.30; H, 5.85; N, 8.11; S, 6.19; Cl, 7.19. Found: C, 60.28; H, 5.87;
N, 8.12; S, 6.16; Cl, 7.18.
5.1.45. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(4-methoxyphenyl)thiourea hydrochloride (18l)
The title compound was prepared in the same manner as de-
scribed for 12a using 17l instead of 11a as a colorless solid. 69%
yield; mp 208–212 °C (MeOH–EtOH); 1H NMR (DMSO-d6) d:
2.93–3.07 (3H, m), 3.15–3.25 (3H, m), 3.92–4.01 (2H, m), 4.21
(1H, br s), 5.91 (1H, br s), 6.88–6.92 (2H, m), 6.94–6.98 (3H, m),
7.21 (2H, d, J = 8.4 Hz), 7.29–7.37 (4H, m), 7.49 (2H, d, J = 8.4 Hz),
8.76 (1H, br s), 8.87 (1H, br s), 9.84–9.89 (2H, m); MS (FAB) m/z:
452 (MH+). Anal. Calcd for C25H29N3O2SꢀHCl: C, 61.53; H, 6.20; N,
8.61; S, 6.57; Cl, 7.26. Found: C, 61.28; H, 6.20; N, 8.51; S, 6.34;
Cl, 7.38.
5.1.50. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(4-sulfamoylphenyl)thiourea hydrochloride (18q)
The title compound was prepared in the same manner as de-
scribed for 12a using 17q instead of 11a as a colorless solid. 65%
yield; mp 198–200 °C (MeOH–EtOAc); 1H NMR (DMSO-d6) d:
2.90–3.10 (3H, m), 3.15–3.25 (3H, m), 3.94–4.01 (2H, m), 4.15–
4.25 (1H, m), 5.90 (1H, d, J = 5.2 Hz), 6.94–6.98 (3H, m), 7.23–
7.33 (6H, m), 7.51 (2H, d, J = 8.0 Hz), 7.71–7.75 (4H, m), 8.75 (1H,
br s), 8.83 (1H, br s), 10.37 (1H, s), 10.47 (1H, s); MS (FAB) m/z:
501 (MH+). Anal. Calcd for C24H28N4O4S2ꢀHCl: C, 53.67; H, 5.44;
N, 10.43; S, 11.94; Cl, 6.60. Found: C, 53.46; H, 5.44; N, 10.42; S,
11.96; Cl, 6.79.
5.1.46. (S)-1-(4-Fluorophenyl)-3-(4-{2-[(2-hydroxy-3-phenoxy-
propyl)amino]ethyl}phenyl)thiourea hydrochloride (18m)
The title compound was prepared in the same manner as de-
scribed for 12a using 17m instead of 11a as a colorless solid. 63%
yield; mp 211–213 °C (EtOH); 1H NMR (DMSO-d6) d: 2.90–3.10
(3H, m), 3.14–3.28 (3H, m), 3.92–4.02 (2H, m), 4.16–4.26 (1H,
m), 5.89 (1H, br s), 6.94–6.98 (3H, m), 7.14–7.23 (4H, m),
5.1.51. (S)-1-(4-{2-[(2-Hydroxy-3-phenoxypropyl)amino]ethyl}-
phenyl)-3-(4-methylphenyl)thiourea hydrochloride (18r)
The title compound was prepared in the same manner as de-
scribed for 12a using 17r instead of 11a as a colorless powder.

T. Maruyama et al. / Bioorg. Med. Chem. 17 (2009) 5510–5519
5519
60% yield; 1H NMR (DMSO-d6) d: 2.28 (3H, s), 2.90–3.10 (3H, m),
3.17–3.23 (3H, m), 3.94–4.01 (2H, m), 4.17–4.23 (1H, m), 5.90
(1H, d, J = 5.2 Hz), 6.95–6.98 (3H, m), 7.13 (2H, d, J = 8.0 Hz), 7.21
(2H, d, J = 8.0 Hz), 7.31 (2H, d, J = 8.0 Hz), 7.37 (2H, d, J = 8.8 Hz),
7.49 (2H, d, J = 8.8 Hz), 9.93 (1H, s), 9.94 (1H, s); MS (FAB) m/z:
436 (MH+). Anal. Calcd for C25H29N3O2SꢀHClꢀ0.3H2O: C, 62.89; H,
6.46; N, 8.80; S, 6.72; Cl, 7.43. Found: C, 62.98; H, 6.38; N, 9.02;
S, 6.62; Cl, 7.54.
the supernatant (mg/dl) was determined calorimetrically by means
of the glucose oxidase method.
Acknowledgments
The authors are grateful to the staff of the Division of Analytical Sci-
ence Laboratories for elemental analysis and spectral measurements.
References and notes
5.2. Pharmacology
1. Arch, J. R.; Ainsworth, A. T.; Cawthorne, M. A.; Piercy, V.; Sennitt, M. V.; Thod, V.
E.; Wilson, C.; Wilson, S. Nature 1984, 309, 163.
2. Howe, R. Drugs Future 1993, 18, 529.
3. Emorine, L. J.; Marullo, S.; Briend-Sutren, M.-M.; Patey, G.; Tate, K.; Delavier-
Klutchko, C.; Strosberg, A. D. Science 1989, 245, 1118.
4. Emorine, L. J.; Feve, B.; Pairault, J.; Briend-Sutren, M.-M.; Nahmias, C.; Marullo,
S.; Delavier-Klutchko, C.; Strosberg, A. D. Am. J. Clin. Nutr. 1992, 55, 215S.
5. Tate, K. M.; Briend-Sutren, M.-M.; Emorine, L. J.; Delavier-Klutchko, C.; Marullo,
S.; Strosberg, A. D. Eur. J. Biochem. 1991, 196, 357.
6. Yamaguchi, O. Urology 2002, 59, 25.
7. Nomiya, M.; Yamagichi, O. J. Urol. 2003, 170, 649.
8. Yamanishi, T.; Yasuda, K.; Kitahara, S.; Nakai, H.; Yoshida, K.-I.; Iizuka, H.
Neurourol. Urodyn. 2006, 25, 815.
9. Weber, A. E. Annu. Rep. Med. Chem. 1998, 33, 193.
5.2.1. Agonistic activity on human b3-, b2-, and b1-ARs
Human b3-, b2-, and b1-stimulating activities were investigated
using a CHO cell system (cells in which human b3-, b2-, and b1-ARs
were compulsorily expressed were used). The agonistic activity of
the compound (10ꢁ10–10ꢁ4 M) was investigated by incubating 105
cells/well of each of the cells on a 24-well plate and checking the
activity after 2 days’ incubation (subconfluent state) using the pro-
duction of cyclic AMP (cAMP) as an index. The amount of cAMP
produced in each cell (pmol/ml) was measured using a radioimmu-
noassay 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 10ꢁ4 M isoproterenol
was defined as 1.00) for each from the resulting dose–reaction
curve.
10. Weyer, C.; Gautier, J. F.; Danforth, E., Jr. Diabetes Metab. 1999, 25, 11.
11. Cantello, B. C. C.; Smith, S. A. Drugs Future 1991, 16, 797.
12. Bloom, J. D.; Claus, T. H. Drugs Future 1994, 19, 23.
13. Arch, J. R. S.; Wilson, S. Int. J. Obesity 1996, 20, 191.
14. Lipworth, B. J. Br. J. Clin. Pharmacol. 1996, 42, 291.
15. de Souza, C. J.; Burkey, B. F. Curr. Pharm. Des. 2001, 7, 1433.
16. Weber, A. E.; Mathvink, R. J.; Perkins, L.; Hutchins, J. E.; Candelore, M. R.; Tota, L.;
Strander, C. D.; Wyvratt, M. J.; Fisher, M. H. Bioorg. Med. Chem. Lett. 1998, 8, 1101.
17. Weber, A. E.; Ok, H. O.; Alvaro, R. F.; Candelore, M. R.; Cacieri, M. A.; Chiu, S.-H. L.;
Deng, L.; Forrest, M. J.; Hom, G. J.; Hutchins, J. E.; Kao, J.; MacIntyre, D. E.;
Mathvink, R. J.; McLoughlin, D.; Miller, R. R.; Newbold, R. C.; Olah, T. V.; Parmee, E.
R.; Perkins, L.; Stearns, R. A.; Strander, C. D.; Szumiloski, J.; Tang, Y. S.; Tota, L.;
Vicario, P. P.; Wyvratt, M. J.; Fisher, M. H. Bioorg. Med. Chem. Lett. 1998, 8, 2111.
18. Fisher, M. H. U.S. Patent 5,451,677, 1995.
19. Scheuerman, R. A.; Tumelty, D. Tetrahedron Lett. 2000, 41, 6531.
20. Maruyama, T.; Onda, K.; Hayakawa, M.; Seki, N.; Takahashi, T.; Moritomo, H.;
Suzuki, T.; Matsui, T.; Takasu, T.; Nagase, I.; Ohta, M. Bioorg. Med. Chem. 2009,
17, 3283.
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 levels under fed
conditions, and then randomly classified into groups. The test com-
pound 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 = 6). Blood samples were collected
from the tail vein using a heparin-treated glass capillary tube after
which the blood was deproteinized, and the amount of glucose in