Design, synthesis, and structure-activity relationships of a series of novel N-aryl-2-phenylcyclopropanecarboxamide that are potent and orally active orexin receptor antagonists

Article abstract of DOI:10.1016/j.bmc.2014.08.034

Herein we describe the design, synthesis, and structure-activity relationships (SARs) of a novel phenylcyclopropane series represented by 7 and 33b as antagonists of orexin 1 and orexin 2 receptors. With 4 serving as the initial lead for the development o

Full text of DOI:10.1016/j.bmc.2014.08.034

Bioorganic & Medicinal Chemistry 22 (2014) 6071–6088
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Design, synthesis, and structure–activity relationships of a series
of novel N-aryl-2-phenylcyclopropanecarboxamide that are potent
and orally active orexin receptor antagonists
a
a
a
a
Yu Yoshida a, , Taro Terauchi , Yoshimitsu Naoe , Yuji Kazuta , Fumihiro Ozaki ,
Carsten T. Beuckmann b, Makoto Nakagawa b, Michiyuki Suzuki b, Ikuo Kushida c, Osamu Takenaka d,
Takashi Ueno d, Masahiro Yonaga a
⇑
a Medicinal Chemistry, Eisai Product Creation Systems, Eisai Co., Ltd, 5-1-3 Tokodai, Tsukuba-shi, Ibaraki 300-2635, Japan
b Biopharmacology, Eisai Product Creation Systems, Eisai Co., Ltd, 5-1-3 Tokodai, Tsukuba-shi, Ibaraki 300-2635, Japan
c Physical Chemistry, Eisai Product Creation Systems, Eisai Co., Ltd, 5-1-3 Tokodai, Tsukuba-shi, Ibaraki 300-2635, Japan
d Drug Metabolism and Pharmacokinetics, Eisai Product Creation Systems, Eisai Co., Ltd, 5-1-3 Tokodai, Tsukuba-shi, Ibaraki 300-2635, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
Herein we describe the design, synthesis, and structure–activity relationships (SARs) of a novel phenyl-
cyclopropane series represented by 7 and 33b as antagonists of orexin 1 and orexin 2 receptors. With 4
serving as the initial lead for the development of orexin antagonists, exploration of SAR resulted in
improved binding affinity for orexin 1 and orexin 2 receptors. Among the synthesized compounds, 33b
((ꢀ)-N-(5-cyanopyridin-2-yl)-2-[(3,4-dimethoxyphenyl)oxymethyl]-2-phenylcyclopropanecarboxamide)
exhibited potent in vitro activity and oral efficacy in animal sleep measurement experiments. The results
of our study suggest that compound 33b may serve as a valuable template for the development of new
orexin receptor antagonists.
Received 16 June 2014
Revised 29 August 2014
Accepted 30 August 2014
Available online 8 September 2014
Keywords:
Orexin receptor antagonist
Hypocretin
Ó 2014 Elsevier Ltd. All rights reserved.
Insomnia
Sleep disorder
1. Introduction
studies have indicated that the orexin system plays a key role in
regulating the sleep/wake cycle.4,5 Abnormalities in orexin signal-
Orexin/hypocretin receptors (OXRs) are a Class A of G-protein-
coupled receptors (GPCRs) that mediate the action of two endoge-
nous neuropeptide ligands known as orexin A and orexin B (33
and 28 amino acids long, respectively), which are also known as
hypocretin 1 and hypocretin 2. Within the central nervous system,
these peptides are exclusively produced in a small cluster of neu-
rons in the posterior and lateral hypothalamus, from where they
project widely throughout the central nervous system.1–3 The orex-
in signaling system is involved in multiple functions related to feed-
ing behavior, metabolism, sleep/arousal states, and brain reward
mechanisms. Indeed, a number of genetic and pharmacological
ing, such as orexin peptide deficiency or a lack of OXRs, produce a
narcolepsy–cataplexy-like phenotype in mice.6,7 Moreover, narco-
lepsy–cataplexy syndrome in humans has been associated with
diminished levels of orexin-A in the cerebrospinal fluid and with
a near or total loss of orexinergic neurons.8–10 There are also reports
that blocking the orexin/hypocretin receptor 2 (OX2R) pathway
results in a noticeable sleep phenotype, while blocking the orex-
in/hypocretin receptor 1 (OX1R) pathway produces a moderate
sleep phenotype but contributes to sleep stabilization, suggesting
that both receptors are involved in sleep/wake regulation.11–18
Despite this contextual evidence, the precise contributions of
OX1R and OX2R signaling pathways to various physiological pro-
cesses have not been completely elucidated. Therefore, further
studies are needed to characterize dual orexin antagonists and
selective orexin antagonists for potential use as sleep drugs.
Given the therapeutic potential of the orexin system, orexin
receptor antagonists have been a significant focus of drug discovery
research, primarily for treating insomnia.19–24 The best studied dual
orexin antagonist to date is almorexant (1),25 which was developed
Abbreviations: OX1R, orexin 1 receptor; OX2R, orexin 2 receptor; THF, tetrahy-
drofuran; TBDPS, tert-butyldiphenylsilyl; DMF, N,N-dimethylformamide; DIAD,
diisopropyl azodicarboxylate; TBAF, tetra-n-butylammonium fluoride; DMSO,
dimethyl sulfoxide; TEA, triethylamine; DCM, dichloromethane; HATU, O-(7-
azabenzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate; DIPEA,
N,N-diisopropylethylamine; Bn, benzyl; NMP, N-methyl-2-pyrrolidone; EEG, elec-
troencephalogram; EMG, electromyogram.
⇑
Corresponding author. Tel.: +81 (0)29 847 7395; fax: +81 (0)29 847 4952.
E-mail address: y11-yoshida@hhc.eisai.co.jp (Y. Yoshida).
http://dx.doi.org/10.1016/j.bmc.2014.08.034
0968-0896/Ó 2014 Elsevier Ltd. All rights reserved.

6072
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
O
O
H
N
N
N
N
N
O
O
N
N
O
HN
O
Cl
N
O
O
N
N
S
F
F
F
F
1
3
2
Almorexant
Suvorexant
(MK-4305)
(SB-649868)
(ACT-078573)
Figure 1. Orexin receptor antagonists in late-stage clinical trials.
by Actelion/GlaxoSmithKline for the treatment of insomnia but dis-
continued at the phase III clinical trial stage. Another compound
developed by GlaxoSmithKline26 SB-649868 (2), was evaluated in
phase II clinical trials earlier than almorexant but was not advanced
to a phase III trial. Another dual orexin antagonist developed by
Merck, suvorexant (3), is currently being reviewed by the United
States Food and Drug Administration (FDA) under a New Drug
Application submitted in 2012 (Fig. 1).27,28
O
O
O
O
4
Herein we report the design, synthesis, structure–activity rela-
tionships, and animal model efficacy for a series of novel orally
active orexin receptor antagonists.
OX2R(Ki): 8.7 µM
OX1R(Ki): >10 µM
Figure 2. Initial hit 4.
2. Seed generation of orexin antagonists from trigger
compound
Table 1
The effects of R hydrogen bond donor or acceptor variants on receptor binding
While pursuing another lead discovery program, we identified
compound 4 (Fig. 2) and discovered that it is an antagonist of
OX2R. This finding shifted our focus to optimizing 4 as a potential
trigger for insomnia treatment. We initially investigated the SAR
at the right R position (Table 1) and found that the introduction of
a benzyl ketone (5) led to decreased binding affinity. We next syn-
thesized amine 6, which could be readily prepared from a common
aldehyde intermediate, and we found that 6 binds to OX2R with 2-
fold better affinity than 4. Furthermore, 6 retained OX2R antagonist
activity and gained equipotent antagonist activity against OX1R.
Based on these initial results, we concluded that the distance
between the cyclopropane ring and the phenyl group was not a
determining factor for the lack of inhibitory activity exhibited by
5. In addition, given the fact that published OXR antagonists have
hydrogen bond acceptors (HBA) such as amide or urea groups, we
predicted that the incorporation of a ketone or amide moiety might
enhance receptor binding potency. To test this, we synthesized the
amide 7, which showed remarkably enhanced OX1R and OX2R bind-
ing affinity, roughly three orders of magnitude better than that of 6.
Next, the non-aryl compound 8 was synthesized to evaluate the
importance of the aromatic ring. The relatively poor binding affinity
of 8 indicated that the aryl ring is crucial for receptor binding. The
role of the amide proton was also investigated by introducing sub-
stituents onto the amide nitrogen in analogs 9 and 10. The relatively
poor affinity of these two analogs confirms that in addition to the
HBA character of the amide, the hydrogen bond donor character of
the amide proton is important for receptor binding. Importantly,
the binding affinities for these analogs for human OX1R and OX2R
are comparable to those of OXR antagonists for which proof-of-con-
cept has already been demonstrated in clinical trials. Furthermore,
the ligand lipophilicity efficiencies (LLE) of these ligands, notably
for OX2R, are higher than those of almorexant (1) or SB-649868
(2) (Table 2). Based on these attributes and on the strength of this
SAR, we chose to continue optimizing additional phenylcyclopro-
pane analogues as potential orexin antagonists.
R
O
O
O
Compound
R
In vitro binding affinity
OX2R (Ki, nM) OX1R (Ki, nM)
O
O
4
8654
>10,000
5
6
>10,000
4226
>10,000
4161
N
H
O
O
O
O
7
8
9
7.2
7.2
N
H
7846
4679
2994
7535
N
H
N
N
10
>10,000
>10,000

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6073
Table 2
3. Chemistry
Comparison of LLE among OXR antagonists
Compound
In vitro binding affinity
OX2R (Ki, nM)
ClogPa
LLE for OX2Rb
The general synthetic pathway for the analogs is described in
Scheme 1. Cyclopropane ring formation was accomplished by
reacting a substituted aryl acetonitrile 11 with racemic epichloro-
hydrin to give lactone 12.24 Reduction of the lactone with sodium
borohydride gave cis-diol 13. After protection of 13 with TBDPS-Cl
to give 14, Mitsunobu reaction of the corresponding alcohol with
various phenols led to the production of compounds 15a–g, which
were deprotected using TBAF and oxidized (Swern and Pinnick
oxidations) to afford intermediates 16a–g, 17a–g, and 18a–g.
Almorexant (1)
SB-649868 (2)
Suvorexant (3)
7
4.7
8.5
11.8
7.2
6.09
5.51
4.79
4.82
2.24
2.57
3.14
3.33
a
ClogP value was calculated using Daylight Software ver. 4.94 (Daylight
Chemical Information Systems, Inc., Niguel, CA).
b
LLE was calculated as pKi ꢀ ClogP.
R1
R1
R1
R1
R1
O
Cl
Ar-OH
d
CN
OTBDPS
O
Ar1
OH
OTBDPS
O
HO
HO
a
b
O
c
11
12
13
14
15a-g
11a:R1=H
11b:R1=3,4-OMe
R1
R1
R2
O
O
e
f
g,h
O
O
Ar1
OH
O
Ar1
O
4; R2=Ph
5; R2=Bn
17a-g
16a-g
O
R1
H
Ar1
NH
O
i
15a
3,4-diOMePh
15b 3,4-diOMe
Ph
Ph
O
O
15c
H
6
15d
15e
H
H
4-MeOPh
3-MeOPh
R1
R1
j
k
O
O
Ar1
15f
H
H
4-MeO,3-MeOCH2Ph
3-MeO,4-MeOCH2Ph
R3HN
O
O
Ar1
HO
15g
7,19-33
18a-g
l,m,n
O
N
HN
O
F
O
31
Scheme 1. Reagents and conditions: (a) NaNH2, benzene, 0 °C–RT, 3 h then EtOH, 1 M KOH, reflux, 16.5 h, then concd HCl RT; (b) NaBH4, MeOH–THF, 0 °C–RT; (c) TBDPS-Cl,
imidazole, DMF, ꢀ10 °C–RT; (d) DIAD, PPh3, 0 °C–RT, overnight; (e) TBAF, THF, RT, 1 h; (f) (COCl)2, DMSO, TEA, DCM, ꢀ78 °C–RT, 1 h; (g) R2MgBr, THF, 0 °C–RT, 1.5 h; (h) Dess–
Martin periodinane, DCM, RT, 1 h; (i) aniline, NaB(OAc)3H, DCM, RT, 2 h; (j) 2-methyl-2-butene, NaClO2, NaH2PO4, Acetone–H2O, RT, 2 h; (k) amine, HATU, DIPEA, DMF, 60 °C,
overnight or CCl3COCl, TEA, 1,4-dioxane, RT, then amine; (l) (COCl)2, DCM, cat. DMF, RT, 1 h; (m) 3,4-diamino-1-fluorobenzene, THF, RT to reflux, 1 h; and (n) AcOH, reflux, 2 h.

6074
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
BnO BnO
HO
HO
OH
O
O
O
c
a,b
d
O
O
O
O
34
35
36
37
Scheme 2. Reagents and conditions: (a) BnBr, K2CO3, DMF, 55 °C, 21 h; (b) NaBH4, MeOH, 0 °C–RT, 2 h; (c) MeI, NaH, NMP, 0 °C–RT; and (d) Pd(OH)2, H2 gas, EtOH, RT, 3 h.
BnO
O
BnO
O
HO
O
O
c
O
O
a,b
38
39
40
Scheme 3. Reagents and conditions: (a) NaBH4, MeOH, 0 °C, 1 h; (b) MeI, NaH, NMP, 0 °C–RT, 1.5 h; and (c) Pd(OH)2, H2 gas, EtOH, RT, 2 h.
Grignard reaction of aldehyde 17a followed by treatment with
Dess–Martin periodinane (DMP) afforded ketones 4 and 5. Reduc-
tive amination using aniline with aldehyde 17a afforded the
desired product 6. Amidation of carboxylic acids 18a–g with vari-
ous amines afforded the desired products 7, 19–33. Benzimidazole
analogue 31 was synthesized from carboxylic acid 18a and 3,4-dia-
mino-1-fluorobenzene.
The syntheses of 4-methoxy-3-methoxymethyl phenol (37) and
3-methoxy-4-methoxymethyl phenol (40) are described in Schemes
2 and 3. To synthesize 37, the phenolic group of 34 was benzyl
protected, and the aldehyde was reduced to obtain alcohol 35, which
was subsequently methylated in the presence of MeI/NaH to pro-
duce 36. Removal of the benzyl protecting group yielded compound
37. To synthesize compound 40, the aldehyde of 38 was reduced, fol-
lowed by methylation in the presence of MeI/NaH to give 39.
Removal of the benzyl protecting group yielded compound 40.
mutagenicity risk of the aniline. Therefore B-ring SAR investiga-
tions were performed with the 2-Py C ring of 19.
Removal of the dimethoxy substituent from 19 led to a drastic
reduction in potency (compound 20), suggesting that this substitu-
ent is important for receptor binding. Unexpectedly, switching the
dimethoxy to the A ring (compound 21) did not abrogate binding
potency. This compound should still be able to form three aromatic
interactions, but it would be predicted to have a conformation dif-
ferent from that of 19. Next, we investigated positional variants of
the B ring dimethoxy substituents. Both the 4- and 3-monomethoxy
derivatives (22 and 23) bound more weakly than 19, suggesting
that both of the oxygens and/or methyls are important for the
receptor interaction.
Based on information from the Cambridge Structural Database
(CSD), the conformation of 3,4-diOMe-Ph was predicted to be nearly
planar, with the methyl groups pointing opposite from one another
and the lone pairs on the oxygens oriented in the same direction. To
clarify the relationship between activity and the conformation of the
substituents, compounds 24 and 25 were synthesized, which retain
both oxygens and both methyls but which should have
conformations different from that of the 3,4-dimethoxy parent.
Although 24 had better binding affinity than either the unsubstitut-
ed or monosubstituted analogs, its affinity was still considerably
weaker than that of 19. Compound 25 bound more weakly than both
19 and 24, suggesting that bulky substituents are not tolerated at the
4-position. Collectively, the relative activities of these derivatives
suggest that both the substitution pattern and the conformation of
the 3,4-diOMePh group are important for binding. Consequently,
our SAR evaluation of other positions was made in the context of a
3,4-diOMePh B-ring.
4. Results and discussion
4.1. Pharmacology and in vitro SAR
Our initial SAR evaluation focused on understanding the impor-
tance of the 3,4-diOMe-Ph group. Specifically, we designed several
analogs to determine the importance of HBAs on OX1R/OX2R bind-
ing activity. Table 3 summarizes the results obtained for the deriv-
atives. Notably, although the substitution of a 2-pyridine (2-Py) for
the phenyl C-ring (compound 19) decreased the binding affinity
slightly compared to that of 7, the 2-Py derivative has superior
physicochemical property and is expected to alleviate the
Table 3
Effects of diOMe substitution on biological activity
O
A
HN
C
O
B
Compound
A-ring
B-ring
C-ring
In vitro binding affinity
OX2R (Ki, nM) OX1R (Ki, nM)
7
19
20
21
22
23
24
25
Ph
Ph
Ph
3,4-Dimethoxy phenyl
3,4-Dimethoxy phenyl
Ph
Ph
7.2
36
7.2
2-Py
2-Py
2-Py
2-Py
2-Py
2-Py
2-Py
1068
6841
>10,000
5538
>10,000
747
967
129
588
1330
384
3336
3,4-Dimethoxy phenyl
Ph
Ph
Ph
Ph
Ph
4-Methoxyphenyl
3-Methoxyphenyl
4-Methoxy-3-methoxymethyl phenyl
3-Methoxy-4-methoxymethyl phenyl
772

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6075
Table 4
Effects of amide groups on biological activity and physicochemical properties
O
F
N
R
N
H
O
O
O
O
O
O
31
Compound
R
In vitro binding affinity
OX2R (Ki, nM) OX1R (Ki, nM)
P-gp efflux corrected FRa
Solubility at pH 7.4b
(
lM)
ClogP
7
19
26
27
28
29
30
31
32
33
(+)-33a
(ꢀ)-33b
Aniline
2-Aminopyridine
3-Aminopyridine
4-Aminopyridine
2-Amino-5-methyl thiazole
2-Amino-4-methyl thiazole
3-Amino-5-methyl isoxazole
7.2
36
51
85
35
14
28
27
34
7.2
0.6
0.9
3.4
4.9
2.4
1.5
NT
1.2
1.0
1.1
NT
1.1
2
4.8
4.2
4.2
4.2
4.6
4.6
3.9
5.2
3.9
3.9
3.9
3.9
1068
206
1425
291
832
505
106
103
123
>2000
106
32
NT
NT
<1
11
<1
28
10
22
NT
15
2-Amino-4-cyano pyridine
2-Amino-5-cyano pyridine
2-Amino-5-cyano pyridine
2-Amino-5-cyano pyridine
14
>2000
5.0
NT = not tested.
a
See P-gp transport assay in Section 6.
pH 7.4: Dulbecco’s phosphate-buffered saline (PBS).
b
To increase the binding affinity of 19 while maintaining or
the three aromatic elements are di-OMe-Ph, CF3Ph, and Ph, the
piperidine ring is a spacer, and CF3Ph and di-OMe-Ph form a
U-shaped structure. In 33b, the cyclopropane ring appears to serve
as the spacer that defines the U-shaped (cis)29 conformation of
the di-OMe-Ph and 2-pyridine around the amide moiety.
reducing its lipophilicity, the SAR of the pyridine C ring was
investigated (Table 4). Introduction of a 3- or 4-pyridine moiety
(26 and 27) reduced the lipophilicity with only a marginal reduc-
tion in binding affinity. However, 26 and 27 exhibited significant
P-glycoprotein (P-gp) efflux activity, whereas 19 was not a P-gp
substrate. Compared to 7, 19 also has a lower LLE (3.29 compared
to 3.33) and is 16-fold more soluble. These results encouraged the
investigation of whether the aniline could be replaced by other
heterocycles, such as a thiazole (28 and 29), an isoxazole (30),
or a benzoimidazole (31). Compound 31 was synthesized using
a different approach to avoid having an amide–aniline moiety.
All four of these compounds bind with equal or better affinity
than 19. The five-membered aromatic C rings did, however, have
decreased solubility or higher tendency to show P-gp efflux.
Therefore, we focused primarily on further 2-pyridine explora-
tion. Introduction of a cyano (CN) group on the 2-pyridine ring,
was expected to decrease lipophilicity. Accordingly, compound
33, a racemic mixture with a CN at the 5-position, was one of
the most potent binders identified among this group. After chiral
HPLC purification, the two enantiomers, 33a and 33b, showed
dramatically different binding affinity, with 33b exhibiting potent
binding to OX2R and OX1R and 33a exhibiting little affinity for
either receptor (Ki > 2000 nM). Compound 33b was selected for
further evaluation because it showed the best balance of
in vitro properties among the compounds.
4.3. In vivo characterization
The representative cyclopropane series compound 33b, which
exhibited promising in vitro activity, was evaluated in vivo by
using a mouse model of spontaneous locomotor activity. In this
model, a reduction of motility is considered a surrogate signal for
sleep promotion.31 Movement was quantified by counting infrared
beam breaks in an open field setting (Fig. 4). The results showed
that an oral dose of 100 mg/kg 33b significantly reduced locomotor
activity. At the 100 mg/kg dose, 33b was present in the CSF at a
concentration above the in vitro binding affinity constant (Ki).
(Table 5). Based on these promising results, 33b was selected for
further evaluation via sleep measurements in mice.
The efficacy of the compound to promote sleep in mice was
determined by electroencephalogram (EEG) and electromyogram
(EMG) measurements during the first 3 h of the dark period as pre-
viously described.4,32 Compound 33b and zolpidem as a positive
control were orally administered at doses of 30 and 100 mg/kg
(Figs. 5 and 6). Administration of 33b produced an accelerated
onset of sleep (Figs. 5A and B; 6A) with a significant and
dose-dependent decrease in wakefulness time during the first 3 h
after a 30 or 100 mg/kg dose (Fig. 5D). When administered at doses
of 30 and 100 mg/kg, 33b also caused a significant and dose-
dependent promotion of non-REM sleep, while REM sleep showed
no significant difference. The positive control, a 100 mg/kg oral
dose of zolpidem decreased wakefulness and promoted non-REM
sleep (Fig. 5J and K).33,34 These data demonstrate that 33b shows
a statistically significant sleep-promoting effect in the mouse.
4.2. Molecular simulation
To better understand the conformation of the cyclopropane
analogues, we used Molecular Operating Environment (MOE,
Ryoka System Inc.) to perform a molecular simulation for com-
pound 33b, which we compared to the low-energy conformation
of the dual OX1/OX2R antagonist almorexant (Fig. 3). Conforma-
tion analysis revealed some shared structural features: three aro-
matic moieties that are linked by a spacer with a U-shape
conformation. Although our SAR study suggested that the HBA of
the amide group is important for receptor binding, the simulation
did not reveal a specific role for the HBA groups. For almorexant,
5. Conclusion
We have described a novel series of orexin receptor antagonists
derived from a compound identified in another discovery program.

6076
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
O
O
N
HN
O
F
F
F
1
Almorexant
(ACT-078573)
O
NH
N
O
N
O
O
33b
Figure 3. Overlay of the low-energy conformer of almorexant (1) and the conformer of 33b30 predicted by molecular simulation.
Our optimization effort produced a promising seed compound (7)
that has low nanomolar binding potency for OX2R and OX1R.
Through further optimization, we elucidated the crucial SAR for
each region of the seed compound. Thus, we identified a compound
(33b) that has improved in vitro binding affinity and reduced
lipophilicity compared to those of the seed compound 7. We then
demonstrated that 33b increased sleep time in mice in a dose-
dependent manner. These promising results suggest potential for
further optimization of this series.
6. Experimental section
6.1. Chemistry
6.1.1. General methods
1H and 13C NMR spectra were recorded on a Bruker Avance
spectrometer (operating at 600 MHz) or Varian Model 500 Unity
Inova (operating at 500 MHz), Varian Mercury 400 spectrometer
(operating at 400 MHz). 13C NMR spectra were recorded on a Bru-
ker Avance spectrometer (operating at 150 MHz). Chemical shifts
were calculated in ppm (d) from the residual CHCl3 signal at (dH)
7.26 ppm and (dC) 77.0 ppm in CDCl3. High-resolution mass spectra
(HRMS) were recorded on a ThermoFisherScientific LTQ-Orbitrap
XL spectrometer (using electrospray ionization). The purity of the
biological tested compounds was determined by an analytical
LC–MS or HPLC method and was found to be greater than or equal
to 95% for all compounds. LC–MS analyses were performed using a
Shimadzu LCMS-2010 EV. Column chromatography was carried
out using a Hi-Flash™ column (40 lm, silica gel and NH-silica
gel, Yamazen Corporation). Chemicals and solvents used in the
study were commercially available.
6.1.2. Phenyl-3-oxabicyclo[3.1.0]hexan-2-one (12a)
A suspension of NaNH2 (17.2 g, 440 mmol) in benzene (250 ml)
was slowly added to a solution of phenyl acetonitrile (23.1 ml,
200 mmol) in benzene (50 ml) with cooling in an ice bath. The
ice bath was removed. Then the reaction mixture was warm up
to room temperature and the reaction mixture was stirred for
3 h. The reaction mixture was cooled with ice bath. To this reaction
Figure 4. Effect of 33b in the locomotor activity paradigm (decrease in infrared
beam breaks during the 1 h after dosing). The compound was administered at doses
of 10, 30, and 100 mg/kg (n = 8 each), and differences between groups were
analyzed using one-way ANOVA with Dunnett’s multiple comparison tests (com-
parison with vehicle). ⁄p < 0.05.
Table 5
Mouse pharmacokinetic data for 33b
Compound
In vitro binding affinity (Ki, nM)
Dose
Route
p.o.
Time
(h)
Plasma concentration
(nM)
CSF concentration
(nM)
OX2R
5.0
OX1R
106
(mg/kg)
100
33b
1
3
9900 442
3737 418
245 39
94 24

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6077
Figure 5. Wake time and sleep time measurements after dosing with 33b or zolpidem. (A–C) Vehicle (10% Cremophor EL, 5% DMSO in saline) or 33b (30 or 100 mg/kg) (n = 7
each) were orally administered immediately before lights out (indicated by the arrow), and hourly amounts of (A) wakefulness, (B) non-REM sleep, and (C) REM sleep were
measured. (D–F) Cumulative amounts of (D) wakefulness, (E) non-REM sleep, and (F) REM sleep relative to vehicle for 3 h after dosing with 33b (derived from the time
courses shown in A–C). (G–I) Vehicle (0.5% methylcellulose p.o., n = 4) or zolpidem (10, 30, 100 mg/kg, n = 4 each) were orally administered immediately before lights out
(indicated by the arrow), and hourly amounts of (G) wakefulness, (H) non-REM sleep, and (I) REM sleep were measured. (J–L) Cumulative amounts of (J) wakefulness, (K) non-
REM sleep, and (L) REM sleep relative to vehicle for 3 h after oral dosing with zolpidem. ⁄p < 0.05, ⁄⁄p < 0.01, ⁄⁄⁄p < 0.001 versus vehicle control (one-way ANOVA followed by
Dunnett’s multiple comparison test).
mixture, epichlorohydrin (15.6 ml, 200 mmol) was added drop-
wise. After addition, the reaction mixture was allowed to slowly
warm up to room temperature and the reaction mixture was stir-
red additional 4 h. The reaction mixture was cooled with ice-bath
and a small amount of water was added dropwise. The reaction
mixture was concentrated in vacuo. To the residue, ethanol
(200 ml) and 1 M potassium hydroxide aqueous solution (100 ml)
were added. The reaction solution was heated to reflux for
16.5 h. After cooling the reaction mixture to room temperature,
12 N hydrochloric acid was added (pH of the mixture was about

6078
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
Figure 6. Sleep latency after dosing with 33b or zolpidem. Sleep latency is defined as the reduction in time to sleep onset for a given treatment compared to respective
vehicle treatment for (A) 33b and (B) zolpidem. ⁄p < 0.05, ⁄⁄p < 0.01 versus vehicle control (one-way ANOVA followed by Dunnett’s multiple comparison test). Vehicle and
animal numbers are the same as in Figure 5.
pH1). After the reaction solution was evaporated, water was added
and extract with ethyl acetate. The organic layer was successively
washed with a saturated sodium bicarbonate aqueous solution and
a saturated saline. The organic layer was dried over MgSO4 and
evaporated. The residue was purified by silica gel column chroma-
tography (n-heptane/ethyl acetate) to give lactam 12a (14.5 g,
41.6% yield).
(dd, J = 12.4, 1.6 Hz, 1H), 4.10 (t, J = 12.0 Hz, 1H), 4.20 (dd, J =
12.0, 5.6 Hz, 1H), 7.21–7.46 (m, 10H), 7.7–7.76 (m, 5H). 13C NMR
(150 MHz, CDCl3) d (ppm): 16.8, 19.1, 25.9, 26.6, 26.9, 32.9, 65.4,
67.8, 126.4, 127.7, 127.9, 127.9, 128.3, 128.9, 129.9, 130.0, 132.9,
134.8, 135.6, 135.6, 144.3. HRMS (ESI(+)) Calcd for C27H33O2Si
[M+H]+: 417.2244, Found: 417.2239.
1H NMR (400 MHz, CDCl3) d (ppm): 1.36 (t, J = 4.8 Hz, 1H), 1.64
(dd, J = 7.6, 4.8 Hz, 1H), 2.53–2.57 (m, 1H), 4.27 (d, J = 9.2 Hz, 1H),
4.45 (dd, J = 9.2, 4.8 Hz, 1H), 7.24–7.36 (m, 3H), 7.39–7.42 (m, 2H).
6.1.5. 2-(tert-Butyldiphenylsilyloxymethyl)-1-(3,4-
dimethoxyphenyl)oxymethyl-1-phenylcyclopropane (15a)
To a solution of alcohol 14a (10.29 g, 24.7 mmol) and 3,4-dime-
thoxy phenol (5.71 g, 37.1 mmol), triphenyl phosphine (9.72 g,
37.1 mmol) in THF (90 ml) was added DIAD (7.28 ml, 37 mmol)
slowly dropwise over 20 min at 0 °C. Then was stirred for same
temperature for 15 min. The reaction mixture was allowed to
warm to room temperature and was stirred for 5 days. The reaction
mixture was concentrated in vacuo. The residue was purified by
column chromatography using n-heptane/AcOEt (50:1–10:1, v/v)
to give 15a (11.47 g, 84% yield) as a light yellow oil.
6.1.3. (2-Hydroxymethyl-1-phenylcyclopropyl)methanol (13a)
To a solution of compound 12a (24.7 g, 142 mmol) in THF–
methanol solution (200–100 ml) was added sodium borohydride
(10.7 g, 284 mmol) at 0 °C. And the reaction mixture was stirred
at room temperature for 1 h. Cooling with ice-bath, water was
added to the reaction mixture. Then the reaction mixture was con-
centrated in vacuo and extracted with ethyl acetate. The organic
layer was washed with saturated saline, then dried over MgSO4
and evaporated. The residue was purified by silica gel column chro-
matography (n-heptane/ethyl acetate) to give the title compound
13a (20.5 g, 81% yield) as colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 0.95 (dd, J = 6.0, 5.2 Hz, 1H),
1.09 (s, 9H), 1.20 (dd, J = 8.8, 5.2 Hz, 1H), 1.58–1.66 (m, 1H), 3.77 (s,
3H), 3.82 (s, 3H), 3.84 (dd, J = 8.4, 4.0 Hz, 1H), 4.02 (dd, J = 11.6,
6.0 Hz, 1H), 4.10 (d, J = 9.6 Hz, 1H), 4.18 (d, J = 9.6 Hz, 1H), 6.26
(dd, J = 8.4, 2.8 Hz, 1H), 6.40 (d, J = 2.8 Hz, 1H), 6.71 (d, J = 8.4 Hz,
1H), 7.19–7.45 (m, 11H), 7.66–7.70 (m, 4H). 13C NMR (150 MHz,
CDCl3) d (ppm): 16.2, 19.3, 26.6, 26.9, 27.4, 29.6, 55.8, 56.5, 63.7,
72.6, 101.1, 104.0, 111.8, 126.3, 127.7, 127.7, 127.7, 128.2, 128.8,
129.6, 129.9, 133.7, 133.7, 135.6, 143.4, 144.4, 149.8, 153.9. HRMS
(ESI(+)) Calcd for C35H41O2Si [M+H]+: 553.2769, Found: 553.2776.
1H NMR (400 MHz, CDCl3) d (ppm): 0.77 (t, J = 5.2 Hz, 1H), 1.08
(dd, J = 8.8, 5.2, 1H), 1.65–1.72 (m, 1H), 2.65 (t, J = 5.6 Hz, 1H), 3.12
(dd, J = 8.4, 2.8 Hz, 1H), 3.42 (td, J = 12.0, 2.8 Hz, 1H), 3.54 (dd,
J = 12.0, 5.2 Hz, 1H), 4.10–4.20 (m, 2H), 7.20–7.25 (m, 1H),
7.28–7.33 (m, 2H), 7.37–7.40 (m, 1H). 13C NMR (150 MHz, CDCl3)
d (ppm):16.8, 25.9, 32.4, 63.6, 67.7, 126.8, 128.5, 129.3, 143.8.
HRMS (ESI(+)) Calcd for C11H18NO2 [M+NH4]+: 196.1332, Found:
196.1336.
6.1.6. 2-Hydroxymethyl-1-(3,4-dimethoxyphenyl)oxymethyl-1-
phenylcyclopropane (16a)
6.1.4. [2-(tert-Butyldiphenylsilyloxymethyl)-1-
phenylcyclopropyl]methanol (14a)
To a solution of compound 15a (1.66 g) in THF (10 ml) was
added tetrabutylammonium fluoride (1 M THF solution: 4.5 ml)
dropwise at room temperature and was stirred at room tempera-
ture for 1 h. Water was added to the reaction solution. Then the
reaction mixture was extracted with ethyl acetate. The organic
layer was washed with a saturated saline, dried over MgSO4, evap-
orated in vacuo. The residue was purified by silica gel column chro-
matography (n-heptane/ethyl acetate) to give 16a as a colorless oil
(890 mg, 94% yield).
To a mixture of 13a (4 g, 22.4 mmol) and imidazole (1.52 g,
22.4 mmol) in DMF (22 mL) was slowly added TBDPS-Cl
(5.82 mL, 22.4 mmol) at ꢀ20 to ꢀ25 °C, and the mixture was stir-
red at the same temperature for 1 h and stirred at 0 °C for 1 h. After
addition of MeOH, the resulting mixture was stirred for 15 min.
The mixture was partitioned between AcOEt and H2O. The organic
layer was washed with saturated NH4Cl solution, brine, dried over
MgSO4, and evaporated. The residue was purified by column chro-
matography (silica gel, n-hexane/AcOEt, 30:1–10:1) to give 14a
(5.53 g, 59.3% yield) as a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 0.92 (dd, J = 6.0, 5.2 Hz, 1H),
1.25 (dd, J = 8.8, 5.2 Hz, 1H), 1.78–1.85 (m, 1H), 2.86 (d, J = 10.8 Hz,
1H), 3.46 (dd, J = 12.0, 10.8 Hz, 1H), 3.81 (s, 3H), 3.82 (s, 3H), 3.91
(d, J = 10.4 Hz, 1H), 4.07–4.13 (m, 1H), 4.52 (d, J = 10.4 Hz, 1H),
6.36 (dd, J = 8.8, 3.2 Hz, 1H), 6.44 (d, J = 3.2 Hz, 1H), 6.73 (d,
J = 8.8 Hz, 1H), 7.20–7.25 (m, 1H), 7.26–7.34 (m, 2H), 7.40–7.43
1H NMR (400 MHz, CDCl3) d (ppm): 0.71 (dd, J = 5.6 Hz, 5.2 Hz,
1H), 1.04 (dd, J = 8.4, 5.2 Hz, 1H), 1.09 (s, 9H), 1.50–1.58 (m, 1H),
3.50 (dd, J = 12.4, 1.6 Hz, 1H), 3.53 (dd, J = 11.6 Hz, 1H), 3.71

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6079
(m, 2H). 13C NMR (150 MHz, CDCl3) d (ppm) 17.2, 27.5, 29.6, 55.9,
was collected, concentrated in vacuo, dried over to give 4 (3 mg,
5.9% yield) as colorless gum.
56.5, 63.5, 74.3, 101.1, 104.7, 111.8, 126.7, 128.3, 128.8, 143.8,
144.2, 149.9, 152.8. HRMS (EI) Calcd for
314.1518, Found: 314.1524.
C
19H22O4 [M]+:
1H NMR (600 MHz, CDCl3) d (ppm) 1.68 (dd, J = 7.6, 4.9 Hz, 1H),
2.15 (dd, J = 6.0, 4.9 Hz, 1H), 3.13 (dd, J = 7.6, 6.4 Hz, 1H), 3.65 (s,
3H), 3.77 (s, 3H), 4.03 (d, J = 9.8 Hz, 1H), 4.41 (d, J = 9.8 Hz, 1H),
6.02 (d, J = 3.0 Hz, 1H), 6.17 (dd, J = 8.9, 2.8 Hz, 1H), 6.63 (d,
J = 8.7 Hz, 1H), 7.33 (d, J = 7.2 Hz, 1H), 7.41 (t, J = 7.74 Hz, 2H),
7.48–7.53 (m, 2H), 7.54–7.62 (m, 3H), 8.04–8.09 (m, 2H). 13C
NMR (150 MHz, CDCl3) d (ppm):19.5, 31.8, 37.6, 55.6, 56.4, 70.4,
100.9, 104.4, 111.6, 127.1, 128.0, 128.4, 128.4, 128.6, 132.6,
138.7, 142.6, 143.5, 149.5, 153.4, 196.8. HRMS (ESI(+)) Calcd for
6.1.7. 2-(3,4-Dimethoxyphenyl)oxymethyl-2-
phenylcyclopropanecarbaldehyde (17a)
To a solution of COCl2 (0.31 mL, 3.6 mmol) in DCM (10 mL) was
added DMSO (0.511 mL, 7.2 mmol) at ꢀ78 °C and stirred for
30 min. A solution of alcohol 16a (0.57 g, 1.8 mmol) in DCM
(5 mL) was added to the reaction mixture at the same temperature
and stirred for 30 min. TEA (2.01 mL, 14.4 mmol) was added to the
reaction mixture and stirred for 15 min. The reaction mixture was
allowed warm up to room temperature. The reaction mixture was
quenched with NH4Claq, extracted with AcOEt, dried over MgSO4,
evapolated in vacuo, and the residue was purified by silica gel col-
umn chromatography using n-heptane/AcOEt (from 4:1 to 3:1, v/v)
to give corresponding aldehyde 17a (558 mg, 99% yield) as a color-
less oil.
C
25H25O4 [M+H]+: 389.1747, Found: 389.1750.
6.1.10. 1-[2-(3,4-Dimethoxyphenoxymethyl)-2-
phenylcyclopropyl]-2-phenylethan-1-one (5)
The title compound was synthesized according to the procedure
described for compound 4 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarbaldehyde 17a and benzyl magne-
sium chloride (7.6% yield) as a colorless gum.
1H NMR (600 MHz, CDCl3) d (ppm): 1.44 (dd, J = 7.6, 4.9 Hz, 1H),
1.87 (dd, J = 6.0, 4.9 Hz, 1H), 2.51 (dd, J = 7.4, 6.2 Hz, 1H), 3.82 (d,
J = 1.1 Hz, 6H), 3.98–4.03 (m, 3H) 4.23 (d, J = 9.8 Hz, 1H), 6.30
(dd, J = 8.7, 2.6 Hz, 1H), 6.38 (d, J = 2.6 Hz, 1H), 6.72 (d, J = 8.7 Hz,
1H), 7.21–7.37 (m, 10H). 13C NMR (150 MHz, CDCl3) d (ppm):
20.7, 34.0, 38.8, 52.8, 56.8, 57.5, 71.2, 102.0, 105.8, 112.9, 128.0,
128.1, 129.3, 129.6, 129.7, 130.9, 135.4, 143.3, 144.6, 150.7,
154.4, 205.8. HRMS (ESI(+)) Calcd for C26H27O4 [M+H]+: 403.1904,
Found: 409.1906.
1H NMR (400 MHz, CDCl3) d (ppm): 1.67 (dd, J = 8.0, 5.2 Hz, 1H),
1.96 (dd, J = 5.2, 5.2 Hz, 1H), 2.37–2.42 (m, 1H), 3.80 (s, 3H), 3.82 (s,
3H), 4.08 (d, J = 10.0 Hz, 1H), 4.42 (d, J = 10.0 Hz, 1H), 6.29 (dd,
J = 8.4, 2.8 Hz, 1H), 6.42 (d, J = 2.8 Hz, 1H), 6.71 (d, J = 8.4 Hz, 1H),
7.26–7.29 (m, 1H), 7.32–7.36 (m, 2H), 7.43–7.45 (m, 2H), 9.70 (d,
J = 4.4 Hz, 1H).
6.1.8. 2-(3,4-Dimethoxyphenyl)oxymethyl-2-
phenylcyclopropanecarboxylic acid (18a)
A mixture of the aldehyde 17a (342 mg, 1.1 mmol) and 2-
methyl-2-butene (0.58 mL, 5.5 mmol) in acetone (10 mL) was
added a mixture of NaH2PO4 (0.13 g, 1.1 mmol) and NaClO2
(0.30 g, 3.3 mmol) in H2O (5 mL) dropwise and stirred at room
temperature for 2.5 h. The reaction mixture was partitioned
between AcOEt and brine, and extracted with AcOEt. The comba-
ined organics was dried over MgSO4, concentrated in vacuo to give
crude carboxylic acid 18a (312 mg, 86.4%) without further
purification.
6.1.11. N-{[2-(3,4-Dimethoxyphenoxymethyl)-2-
phenylcyclopropyl]methyl}aniline (6)
To a solution of aldehyde 17a (31.2 mg, 0.1 mmol) in DCM
(2 ml) was added aniline (10.9 ll, 0.12 mmol) and NaB(OAc)3H
(31.7 mg, 0.15 mmol) at room temperature and being stirred for
2 h. The reaction was quenched with satd NH4Cl. The resulting
mixture was partitioned between AcOEt and satd NaHCO3. The
organic layer was washed with brine, dried over MgSO4, and evap-
orated. The residue was purified by NH-silica gel column chroma-
tography using n-heptane/AcOEt (4:1, v/v) to give amine 6 (32 mg,
82.2% yield) as light brown gum.
1H NMR (400 MHz, CDCl3) d (ppm): 1.60 (dd, J = 8.0, 4.8 Hz, 1H),
1.71 (dd, J = 6.0, 4.8 Hz, 1H), 2.14 (dd, J = 8.0, 6.0 Hz, 1H), 3.79 (s,
3H), 3.80 (s, 3H), 4.24 (t, J = 9.6 Hz, 1H), 4.38 (d, J = 9.6 Hz, 1H),
6.30 (dd, J = 8.4, 2.8 Hz, 1H), 6.42 (d, J = 2.8 Hz, 1H), 6.42 (d,
J = 8.4 Hz, 1H), 7.24–7.27 (m, 1H), 7.31–7.34 (m, 2H), 7.43–7.45
(m, 2H). 13C NMR (150 MHz, CDCl3) d (ppm): 17.2, 27.5, 29.6,
55.9, 56.5, 63.5, 74.3, 101.1, 104.7, 111.8, 126.7, 128.3, 128.8,
143.8, 144.2, 149.9, 152.8. HRMS (ESI(ꢀ)) Calcd for C19H19O5
[MꢀH]ꢀ: 327.1238, Found: 327.1236.
1H NMR (600 MHz, CDCl3) d (ppm): 0.96 (t, J = 5.7 Hz, 1H), 1.26
(dd, J = 8.7, 5.3 Hz, 1H), 1.75–1.82 (m, 1H), 3.07 (dd, J = 12.8, 9.4 Hz,
1H), 3.69 (dd, J = 12.8, 5.7 Hz, 1H), 3.79 (s, 3H) 3.81 (s, 3H), 3.91 (d,
J = 10.2 Hz, 1H), 4.41(br s 1H), 4.48 (d, J = 10.2 Hz, 2H), 6.35 (dd,
J = 8.7, 2.6 Hz, 1H) 6.44 (d, J = 2.6 Hz, 1H), 6.68–6.74 (m, 4H),
7.17–7.26 (m, 3H) 7.32 (t, J = 7.7 Hz, 2H) 7.42 (d, J = 7.3 Hz, 2H).
13C NMR (150 MHz, CDCl3) d (ppm): 17.3, 24.9, 29.6, 44.7, 55.8,
56.5, 73.5, 101.3, 104.7, 111.8, 113.3, 117.6, 126.6, 128.3, 128.7,
129.3, 143.9, 148.5, 149.9, 153.3. HRMS (ESI(+)) Calcd for
6.1.9. [2-(3,4-Dimethoxyphenoxymethyl)-2-
phenylcyclopropyl]phenyl-methanone (4)
C
25H28NO3 [M+H]+: 390.2064, Found: 390.2067.
To a solution of 2-(3,4-dimethoxyphenyl)oxymethyl-2-phenyl-
cyclopropanecarbaldehyde 17a (40.6 mg, 0.13 mmol) in THF was
added phenyl magnesium bromide (1 M THF solution: 0.26 ml,
0.26 mmol) dropwise at 0 °C. The reaction mixture was stirred
for 1.5 h. Then the reaction mixture was quenched with satd NH4Cl
solution. The resulting mixture was poured into H2O. The aqueous
phase was extracted with DCM. The organic layer was washed with
brine, dried over MgSO4, concentrated in vacuo. The residue was
used in the next step without further purification. The residue
was dissolved in DCM and Dess–Martin periodinane (82.7 mg,
0.195 mmol) was added to the reaction mixture at room tempera-
ture. Then the reaction mixture was stirred at room temperature
for 1 h. The resulting mixture was partitioned between DCM and
H2O, and the organic layer was washed with brine, dried over
MgSO4, and concentrated in vacuo. The residue was purified by
LC–MS fractionation. The mass fraction of the target compound
6.1.12. N-Phenyl-2-[3,4-dimethoxyphenyl)oxymethyl]-2-
phenylcyclopropanecarboxamide (7)
To a solution of 2-(3,4-dimethoxyphenyl)oxymethyl-2-phenyl-
cyclopropanecarboxylic acid 18a (32.8 mg, 0.1 mmol) in DMF
(2 ml) was added TEA (41.8 ll, 0.3 mmol) and aniline (18.2 ll,
0.2 mmol) and HBTU (56.9 mg, 0.15 mmol) at room temperature.
The reaction mixture was stirred for 2 h. Then The reaction mixture
was partitioned between AcOEt and H2O. The organic layer was
washed with brine, dried with MgSO4, and evaporated. The residue
was purified by NH-silica gel column chromatography using
n-heptane/AcOEt (4:1, v/v) to give amide 7 (30 mg, 75% yield) as
a white solid.
1H NMR (400 MHz, CDCl3) d (ppm): 1.54–1.59 (m, 1H), 1.87 (t,
J = 5.6 Hz, 1H), 2.04–2.10 (m, 1H), 3.62 (s, 3H), 3.77 (s, 3H), 4.25
(d, J = 9.6 Hz, 1H), 4.53 (d, J = 9.6 Hz, 1H), 6.32–6.34 (m, 2H),

6080
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6.65–6.68 (m, 1H), 7.05–7.09 (m, 1H), 7.25–7.29 (m, 3H),
7.33–7.36 (m, 2H), 7.43–7.47 (m, 4H), 7.64 (br s, 1H). 13C NMR
(150 MHz, CDCl3) d (ppm): 18.6, 30.4, 34.0, 55.6, 56.4, 71.2,
101.5, 105.0, 111.7, 119.9, 124.2, 127.0, 127.9, 128.5, 129.0,
138.0, 142.5, 143.7, 149.6, 153.5, 168.0. HRMS (ESI(+)) Calcd for
mixture, epichlorohydrin (9.26 ml, 119 mmol) in toluene (10 ml)
was added dropwise. After addition, the reaction mixture was
allowed to slowly warm up to room temperature and the reaction
mixture was stirred additional 15 h. The reaction mixture was
cooled with ice-bath and a small amount of water was added drop-
wise. The reaction mixture was concentrated in vacuo. The residue
was dissolved in ethanol (100 ml) and water (100 ml) and potas-
sium hydroxide (13.3 g, 237 mmol) was added. The reaction solu-
tion was heated to reflux for 15 h. The reaction mixture was
concentrated in vacuo to remove ethanol. The solution was parti-
tioned with toluene. After adding concd HCl (pH of the mixture
was pH 2) to the aqueous layer, toluene (200 ml) was added, then
was heated to 70 °C and stirred for 3 h. The organic layer was
partitioned. Then aqueous layer was extracted with EtOAc. The
combined organic layer was successively washed with a satd
NaHCO3 solution and a saturated saline. The organic layer was
dried over MgSO4 and evaporated. The residue was purified by sil-
ica gel column chromatography (n-heptane/ethyl acetate) to give
12b (17.6 g, 63.2% yield) as an orange oil.
C
25H26N2O4 [M+H]+: 404.1856, Found: 404.1858.
6.1.13. N-Cyclohexyl-2-[3,4-dimethoxyphenyl)oxymethyl]-2-
phenylcyclopropanecarboxamide (8)
The title compound was synthesized according to the procedure
described for compound 4 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid 18a and cyclohexyl
amine 8 (93% yield) as a white solid.
1H NMR (600 MHz, CDCl3) d (ppm): 1.10–1.21 (m, 2H), 1.23–
1.41 (m, 3H), 1.44 (dd, J = 8.3, 4.9 Hz, 1H), 1.59–1.99 (m, 7H),
3.82 (s, 7H), 4.26 (d, J = 9.4 Hz, 1H), 4.44 (d, J = 9.4 Hz, 1H), 5.68
(d, J = 7.9 Hz, 1H), 6.36 (dd, J = 8.7, 2.6 Hz, 1H), 6.43 (d, J = 2.6 Hz,
1H), 6.73 (d, J = 8.7 Hz, 1H), 7.26 (d, J = 7.2 Hz, 1H), 7.34 (t,
J = 7.7 Hz, 2H), 7.40 (d, J = 7.2 Hz, 2H). 13C NMR (150 MHz, CDCl3)
d (ppm): 18.2, 24.8, 24.9, 25.6, 29.5, 32.8, 33.2, 33.3, 48.6, 55.8,
56.5, 70.5, 101.1, 104.6, 111.9, 126.8, 127.9, 128.4, 142.8, 143.5,
149.6, 153.6, 168.6. HRMS (ESI(+)) Calcd for C25H32NO4 [M+H]+:
410.2326, Found: 410.2327.
1H NMR (400 MHz, CDCl3) d (ppm): 1.34 (t, J = 4.8 Hz, 1H), 1.62
(dd, J = 8.0, 4.8 Hz, 1H), 2.50–2.55 (m, 1H), 3.87 (s, 3H), 3.91 (s, 3H),
4.29 (d, J = 9.2 Hz, 1H), 4.48 (dd, J = 9.2, 4.4 Hz, 1H), 6.83 (d,
J = 8.0 Hz, 1H), 6.88 (dd, J = 8.0, 2.8 Hz, 1H), 7.03 (d, J = 2.8 Hz,
1H). 13C NMR (150 MHz, CDCl3) d (ppm): 20.1, 25.0, 31.7, 56.0,
68.1, 111.2, 112.2, 120.5, 126.6, 148.8, 149.0, 176.3. HRMS (EI)
Calcd for C13H14O4 [M]+: 234.0892, Found: 234.0898.
6.1.14. N-Methyl-N-phenyl-2-[(3,4-dimethoxyphenyl)oxymethyl]-
2-phenylcyclopropanecarboxamide (9)
The title compound was synthesized according to the procedure
described for compound 4 from 2-(3,4-dimethoxyphenyl)oxymeth-
yl-2-phenylcyclopropanecarboxylic acid 18a and N-methylaniline
(61.3% yield) as a colorless gum.
6.1.17. [1-(3,4-Dimethoxyphenyl)-2-
(hydroxymethyl)cyclopropyl]methanol (13b)
To
a suspension of lithium aluminum hydride (311 mg,
1H NMR (600 MHz, CDCl3) d (ppm): 1.25 (dd, J = 7.4, 4.7 Hz, 1H),
1.88 (t, J = 5.1 Hz, 1H), 1.98 (t, J = 6.6 Hz, 1H), 3.31 (s, 3H), 3.81–
3.87 (m, 6H), 4.30 (d, J = 9.8 Hz, 1H), 4.41 (d, J = 9.8 Hz, 1H) 6.42
(dd, J = 8.7, 2.6 Hz, 1H) 6.48 (d, J = 2.6 Hz, 1H) 6.76 (d, J = 8.7 Hz,
1H) 7.06 (d, J = 6.4 Hz, 2H) 7.12–7.21 (m, 3H) 7.32–7.55 (m, 5H).
13C NMR (150 MHz, CDCl3) d (ppm): 19.0, 26.8, 34.2, 37.8, 55.9,
56.5, 71.3, 101.1, 105.5, 112.1, 126.6, 127.6, 127.8, 128.1, 128.2,
129.6, 142.6, 143.6, 144.2, 149.7, 153.8, 169.8. HRMS (ESI(+)) Calcd
for C26H28NO4 [M+H]+: 418.2013, Found: 418.2018.
8.2 mmol) in THF (20 ml) was added solution of compound 12b
(1.92 g, 8.2 mmol) in THF (10 ml) dropwise at 0 °C. And the
reaction mixture was stirred at same temperature for 1 h. Small
amount of water (0.31 ml), followed by aqueous 15% NaOH solu-
tion (0.31 ml), water (0.93 ml) was added to the reaction mixture.
Then the reaction mixture was added celite, and was filtered. The
filtrate was concentrated in vacuo. The residue was purified by sil-
ica gel column chromatography using n-heptane/AcOEt (2:1–2:3,
v/v) to give alcohol 13b (1.49 g, 76.3% yield) as pale yellow oil.
1H NMR (400 MHz, CDCl3) d (ppm): 0.75 (t, J = 5.2 Hz, 1H), 1.06
(dd, J = 8.4, 5.2 Hz, 1H), 1.64–1.73 (m, 1H), 2.47–2.53 (m, 1H), 3.05–
3.10 (m, 1H), 3.41 (t, J = 11.6 H, 1H), 3.54 (dd, J = 11.6, 5.2 Hz, 1H),
3.86 (s, 3H), 3.89 (s, 3H), 4.10–4.22 (m, 2H), 6.82 (d, J = 8.0 Hz, 1H),
6.93–6.97 (m, 2H). 13C NMR (150 MHz, CDCl3) d (ppm): 16.8, 25.9,
32.3, 56.0, 63.7, 67.9, 111.2, 112.7, 121.6, 136.4, 148.1, 148.9.
HRMS (ESI(+)) Calcd for C13H22NO4 [M+NH4]+: 256.1543, Found:
256.1539.
6.1.15. 1-[2-(3,4-Dimethoxyphenoxymethyl)-2-
phenylcyclopropanecarbonyl]-1,2,3,4-tetrahydroquinoline (10)
The title compound was synthesized according to the procedure
described for compound 4 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid 18a and 1,2,3,4-tetra-
hydroqunoline (93% yield) as a white amorphous solid.
1H NMR (600 MHz, CDCl3) d (ppm): 1.38–1.49 (m, 1H), 1.83 (br
s, 1H), 2.01 (t, J = 5.3 Hz, 1H), 2.11 (dt, J = 12.9, 6.2 Hz, 1H), 2.37 (dd,
J = 7.7, 6.2 Hz, 1H), 2.65–2.71 (m, 1H), 2.77 (dt, J = 6.1, 15.7 Hz, 1H),
3.42 (br s, 1H), 3.72–3.84 (m, 6H), 4.18–4.29 (m, 1H), 4.33 (br s,
1H), 4.57 (br s, 1H), 6.37–6.46 (m, 2H), 6.73 (d, J = 8.3 Hz, 1H),
6.95 (br s, 1H), 7.05 (t, J = 7.4 Hz, 1H), 7.14 (d, J = 7.6 Hz, 1H),
7.20–7.25 (m, 1H), 7.30 (d, J = 6.4 Hz, 4H), 7.65 (d, J = 7.2 Hz, 1H).
13C NMR (150 MHz, CDCl3) d (ppm): 20.3, 24.0, 24.2, 27.0, 29.3,
29.7, 33.3, 43.5, 55.8, 56.5, 70.5, 101.1, 105.3, 112.0, 125.0, 126.0,
126.6, 126.9, 128.2, 128.4, 128.6, 133.3, 139.0, 142.0, 143.7,
149.7, 153.7, 169.4. HRMS (ESI(+)) Calcd for C28H30NO4 [M+H]+:
444.2169, Found: 444.2171.
6.1.18. (2-{[(tert-Butyldiphenylsilyl)oxy]methyl}-1-(3,4-
dimethoxyphenyl)cyclopropyl)methanol (14b)
To a mixture of 13b (1.49 g, 6.25 mmol) and imidazole (425 mg,
6.25 mmol) in DMF (10 mL) was slowly added solution of TBDPS-Cl
(15.2 mL, 6.25 mmol) in DMF (3 ml) at 0 °C, and the mixture was
stirred at the same temperature for 1 h. The reaction mixture
was allowed to warm up to room temperature, then was sitrred
for 2 h. After addition of MeOH, the resulting mixture was stirred
for 15 min. The reaction mixture was partitioned between AcOEt
and H2O. The organic layer was washed with water, brine, dried
over MgSO4, and evaporated. The residue was purified by silica
gel column chromatography using n-heptane/AcOEt (5:1–3:1,
v/v) to give 14b (1.5 g, 50.5% yield) as a colorless oil.
6.1.16. 1-(3,4-Dimethoxyphenyl)-3-oxabicyclo[3.1.0]hexan-2-
one (12b)
To a suspension of sodium hydride in NMP-toluene (90–30 ml)
at 0 °C, 3,4-dimethoxyphenyl acetonitrile (21 g, 119 mmol) was
slowly added dropwise. The ice bath was removed, then was warm
up to room temperature and the reaction mixture was stirred for
6 h. The reaction mixture was cooled with ice bath. To this reaction
1H NMR (400 MHz, CDCl3) d (ppm): 0.68 (t, J = 5.2 Hz, 1H), 1.13
(dd, J = 8.8, 5.2 Hz, 1H), 1.11 (s, 9H), 1.47–1.56 (m, 1H), 3.45 (dd,
J = 11.2, 2.0 Hz, 1H), 3.52 (t, J = 11.2 Hz, 1H), 3.71 (dd, J = 11.2,
1.6 Hz, 1H), 3.88 (s, 6H), 4.03 (t, J = 11.6 Hz, 1H), 4.19 (dd,
J = 11.6, 5.2 Hz, 1H), 6.83 (d, J = 8.4 Hz, 1H), 6.95 (dd, J = 8.4,

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6081
2.0 Hz, 1H), 7.04 (d, J = 2.0 Hz, 1H), 7.35–7.49 (m, 6H), 7.69–7.77
(m, 4H). 13C NMR (150 MHz, CDCl3) d (ppm): 16.5, 19.1, 25.9,
26.8, 32.8, 55.8, 56.0, 65.3, 68.2, 111.1, 112.8, 121.0, 127.9, 127.9,
123.0, 130.0, 132.9, 132.9, 135.4, 135.5, 135.6, 137.1, 147.8,
148.7. HRMS (ESI(+)) Calcd for C29H37O4Si [M+H]+: 477.2456,
Found: 477.2452.
9.87 mmol), NaH2PO4 (236 mg, 1.97 mmol) and NaClO2
(533 mg, 5.9 mmol) and stirred at room temperature for 2 h.
The reaction mixture was quenched with satd NH4Cl and parti-
tioned between AcOEt and water. The organic layer was washed
with water, aq Na2S3O3, brine, and dried over MgSO4. Then was
concentrated in vacuo. The resulting solid was washed with
Et2O-n-Hep to give carboxylic acid 18b (568 mg, 87.8% yield) as
a white solid.
6.1.19. tert-Butyl({[2-(3,4-dimethoxyphenyl)-2-
(phenoxymethyl)cyclopropyl]methoxy})diphenylsilane (15b)
The title compound was synthesized according to the procedure
described for compound 15a from (2-{[(tert-butyldiphenylsi-
lyl)oxy]methyl}-1-(3,4-dimethoxyphenyl)cyclopropyl)methanol
(14b) and phenol (89.6% yield) as a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 1.58 (dd, J = 8.4, 5.6 Hz, 1H),
1.69 (t, J = 5.6 Hz, 1H), 2.11 (dd, J = 8.4, 5.6 Hz, 1H), 3.86 (s, 3H),
3.89 (s, 3H), 4.27 (d, J = 9.6 Hz, 1H), 4.37 (d, J = 9.6 Hz, 1H), 6.77–
6.84 (m, 3H), 6.87–6.92 (m, 1H), 6.97 (dd, J = 8.0, 2.0 Hz, 1H),
7.02 (d, J = 2.0 Hz, 1H), 7.18–7.24 (m, 3H). 13C NMR (150 MHz,
CDCl3) d (ppm): 20.1, 25.5, 35.8, 55.9, 56.0, 70.6, 110.9, 112.6,
114.8, 120.8, 120.9, 129.3, 134.6, 148.4, 148.7, 159.0, 176.1. HRMS
(ESI(ꢀ)) Calcd for C19H19O5 [MꢀH]ꢀ: 327.1238, Found: 327.1234.
1H NMR (600 MHz, CDCl3) d (ppm): 0.90 (t, J = 5.5 Hz, 1H), 1.11
(s, 9H), 1.18 (dd, J = 8.9, 5.1 Hz, 1H), 1.58–1.65 (m, 1H), 3.78 (dd,
J = 11.1, 8.9 Hz, 1H), 3.87 (m, 6H), 4.07 (dd, J = 11.1, 5.9 Hz, 1H),
4.13 (d, J = 9.8 Hz, 1H), 4.18–4.23 (m, 1H), 6.76–6.82 (m, 3H),
6.92 (t, J = 7.4 Hz, 1H), 6.98 (dd, J = 8.1, 2.1 Hz, 1H), 7.05 (d,
J = 1.9 Hz, 1H), 7.24 (dd, J = 8.5, 7.4 Hz, 2H), 7.30–7.47 (m, 6H),
7.66–7.75 (m, 4H). 13C NMR (150 MHz, CDCl3) d (ppm): 15.8,
19.2, 26.9, 27.2, 29.6, 55.8, 55.9, 63.7, 72.3, 110.9, 112.9, 114.7,
120.5, 121.1, 127.7, 129.3, 129.6, 133.7, 133.8, 135.6, 137.2,
6.1.23. tert-Butyl({[2-(phenoxymethyl)-2-phenylcyclopropyl]
methoxy})diphenylsilane (15c)
The title compound was synthesized according to the procedure
described for compound 3 from [2-(tert-butyldiphenylsilyloxym-
ethyl)-1-phenylcyclopropyl]methanol
(84.6% yield) as a colorless oil.
14a
and
phenol
147.7, 148.5, 159.1. HRMS (ESI(+)) Calcd for
C35H44NO4Si
[M+NH4]+: 570.3034, Found: 570.3039.
1H NMR (400 MHz, CDCl3) d (ppm): 0.90–0.96 (m, 1H), 1.08 (s,
9H), 1.17–1.22 (m, 1H), 1.57–1.66 (m, 1H), 3.82 (dd, J = 11.2,
8.4 Hz, 1H), 4.02 (dd, J = 11.2, 6.0 Hz, 1H), 4.16 (d, J = 10.0 Hz, 1H),
4.20 (d, J = 10.0 Hz, 1H), 6.75–6.80 (m, 2H), 6.87–6.92 (m, 1H),
7.17–7.44 (m, 13H), 7.65–7.70 (m, 4H). 13C NMR (150 MHz, CDCl3)
d (ppm):19.0, 26.8, 34.2, 37.8, 55.9, 56.5, 71.3, 101.1, 105.5, 112.1,
126.6, 127.6, 127.8, 128.1, 128.2, 129.6, 142.6, 143.6, 144.2, 149.7,
6.1.20. (2-(3,4-Dimethoxyphenyl)-2-
(phenoxymethyl)cyclopropyl)methanol (16b)
The title compound was synthesized according to the procedure
described for compound 16a from (2-{[(tert-butyldiphenylsi-
lyl)oxy]methyl}-1-(3,4-dimethoxyphenyl)cyclopropyl)methanol
(15b) as a colorless oil. (78.1% yield).
153.8, 169.8. HRMS (ESI(+)) Calcd for
493.2557, Found: 493.2562.
C
33H37O2Si [M+H]+:
1H NMR (400 MHz, CDCl3) d (ppm): 0.88 (t, J = 5.2 Hz, 1H), 1.23
(dd, J = 8.8, 5.2 Hz, 1H), 1.74–1.83 (m, 1H), 2.83 (dd, J = 10.8, 2.0 Hz,
1H), 3.40–3.48 (m, 1H), 3.85 (s, 3H), 3.87 (s, 3H), 3.93 (d,
J = 10.0 Hz, 1H), 4.05–4.13 (m, 1H), 4.52 (d, J = 10.0 Hz, 1H), 6.79
(d, J = 8.0 Hz, 1H), 6.84–6.88 (m, 2H), 6.93–7.01 (m, 3H), 7.23–
7.28 (m, 2H). 13C NMR (150 MHz, CDCl3) d (ppm): 16.8, 27.4,
29.5, 56.0, 63.5, 73.9, 111.0, 112.6, 114.9, 121.1, 121.6, 129.6,
136.5, 147.9, 148.7, 158.2. HRMS (ESI(+)) Calcd for C19H26NO4
[M+NH4]+: 332.1856, Found: 332.1856.
6.1.24. tert-Butyl({[2-(4-methoxyphenoxymethyl)-2-phenyl-
cyclopropyl]methoxy})diphenylsilane (15d)
The title compound was synthesized according to the procedure
described for compound 3 from [2-(tert-butyldiphenylsilyloxym-
ethyl)-1-phenylcyclopropyl]methanol 14a and 4-methoxyphenol
(64.8% yield) as a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 0.90–0.96 (m, 1H), 1.08 (s,
9H), 1.18 (dd, J = 8.8, 5.2 Hz, 1H), 1.57–1.64 (m, 1H), 3.75 (s, 3H),
3.83 (dd, J = 11.6, 8.4 Hz, 1H), 4.00 (dd, J = 11.6, 5.6 Hz, 1H), 4.11
(d, J = 10.0 Hz, 1H), 4.14 (d, J = 10.0 Hz, 1H), 6.67–6.72 (m, 2H),
6.74–6.79 (m, 2H), 7.17–7.44 (m, 11H), 7.65–7.71 (m, 4H). 13C
NMR (150 MHz, CDCl3) d (ppm): 16.2, 19.2, 26.9, 27.3, 29.7, 55.8,
63.7, 72.8, 114.5, 115.7, 126.3, 127.7, 127.7, 128.2, 128.9, 129.6,
133.8, 133.8, 135.6, 144.5, 153.4, 153.7. HRMS (ESI(+)) Calcd for
6.1.21. 2-(3,4-Dimethoxyphenyl)-2-
(phenoxymethyl)cyclopropanecarbaldehyde (17b)
To a solution of COCl2 (0.38 mL, 4.44 mmol) in DCM (6 mL) was
added DMSO (0.63 mL, 8.86 mmol) at ꢀ78 °C and stirred for 5 min.
The above-mentioned alcohol 16b (697 mg, 2.22 mmol) in DCM
(7 mL) was added to the reaction mixture then was stirred for
30 min. TEA (2.47 mL, 17.7 mmol) was added to the reaction mix-
ture at same temperature and stirred for 15 min and then the reac-
tion mixture was allowed to warm up to room temperature. After
stirred for 45 min, the reaction mixture was quenched with satd
NH4Claq, extracted with AcOEt, washed with water, satd NaHCO3,
brine, dried over MgSO4, concentrated in vacuo, and the residue
was purified by silica gel column chromatography using n-hep-
tane/AcOEt (5:1–3:1, v/v) to give aldehyde 17b (615 mg,
88.7% yield) as a colorless oil.
C
34H39O3Si [M+H]+: 523.2663, Found: 523.2669.
6.1.25. tert-Butyl({[2-(3-methoxyphenoxymethyl)-2-phenyl-
cyclopropyl]methoxy})diphenylsilane (15e)
The title compound was synthesized according to the procedure
described for compound 3 from [2-(tert-butyldiphenylsilyloxym-
ethyl)-1-phenylcyclopropyl]methanol 14a and 3-methoxyphenol
(76% yield) as a light yellow oil.
1H NMR (400 MHz, CDCl3) d (ppm): 0.90–0.96 (m, 1H), 1.08 (s,
9H), 1.19 (dd, J = 8.8, 4.8 Hz, 1H), 1.57–1.65 (m, 1H), 3.74 (s, 3H),
3.81 (dd, J = 11.6, 8.4 Hz, 1H), 4.01 (dd, J = 11.6, 6.0 Hz, 1H), 4.16
(d, J = 10.0 Hz, 1H), 4.19 (d, J = 10.0 Hz, 1H), 6.35–6.40 (m, 2H),
6.45–6.49 (m, 1H), 7.09–7.14 (m, 1H), 7.17–7.44 (m, 11H), 7.65–
7.71 (m, 4H). 13C NMR (150 MHz, CDCl3) d (ppm): 16.2, 19.2,
26.9, 27.3, 29.5, 55.2, 63.7, 72.0, 101.1, 106.5, 106.7, 126.3, 127.7,
127.7, 128.2, 128.9, 129.6, 129.7, 133.7, 133.7, 135.6, 144.4,
1H NMR (400 MHz, CDCl3) d (ppm): 1.68 (dd, J = 5.2, 8.0 Hz, 1H),
1.94 (t, J = 5.2 Hz, 1H), 2.13–2.39 (m, 1H), 3.86 (s, 3H), 3.89 (s, 3H),
4.12 (d, J = 10.0 Hz, 1H), 4.41 (d, J = 10.0 Hz, 1H), 6.79–6.84 (m, 3H),
6.90–6.95 (m, 1H), 6.97 (dd, J = 8.4, 2.0 Hz, 1H), 7.02 (d, J = 2.0 Hz,
1H), 7.20–7.27 (m, 2H), 9.68 (d, J = 3.6 Hz, 1H).
6.1.22. 2-(3,4-Dimethoxyphenyl)-2-
(phenoxymethyl)cyclopropane-1-carboxylic acid (18b)
To a solution of the aldehyde 17b (615 mg, 1.97 mmol) in ace-
tone–water (12–6 ml) was added 2-methyl-2-butene (1.04 mL,
160.4, 160.7. HRMS (ESI(+)) Calcd for
523.2663, Found: 523.2663.
C
34H39O3Si [M+H]+:

6082
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6.1.26. tert-Butyl({2-[3-methoxy-4-(methoxymethyl)phenoxy-
methyl]-2-phenylcyclopropyl}methoxy)diphenylsilane (15f)
The title compound was synthesized according to the procedure
described for compound 3 from [2-(tert-butyldiphenylsilyloxym-
ethyl)-1-phenylcyclopropyl]methanol 14a and phenol 40
(76.9% yield) as a light yellow oil.
6.1.30. [2-(3-Methoxyphenoxymethyl)-2-phenylcyclopropyl]
methanol (16e)
The title compound was synthesized according to the procedure
described for compound 16a from tert-butyl({[2-(3-methoxyphen-
oxymethyl)-2-phenylcyclopropyl]methoxy})diphenylsilane
(88% yield) as a colorless oil.
15e
1H NMR (400 MHz, CDCl3) d (ppm): 0.95 (dd, J = 6.0, 5.2 Hz, 1H),
1.08 (s, 9H), 1.20 (dd, J = 8.8, 5.2 Hz, 1H), 1.58–1.66 (m, 1H), 3.35 (s,
3H), 3.73 (s, 3H), 3.81 (dd, J = 11.2, 8.4 Hz, 1H), 4.03 (dd, J = 11.2,
6.0 Hz, 1H), 4.18 (d, J = 10.0 Hz, 1H), 4.23 (d, J = 10.0 Hz, 1H), 4.40
(s, 2H), 6.33 (dd, J = 8.0, 2.4 Hz, 1H), 6.35 (d, J = 2.4 Hz, 1H), 7.15
(d, J = 8.0 Hz, 1H), 7.18–7.24 (m, 1H), 7.26–7.44 (m, 10H), 7.65–
7.71 (m, 4H). 13C NMR (150 MHz, CDCl3) d (ppm): 17.2, 20.2,
27.9, 28.4, 30.5, 56.4, 58.8, 64.7, 70.4, 73.1, 100.1, 105.8, 119.7,
127.3, 128.7, 128.7, 129.2, 129.8, 130.6, 131.3, 134.7, 134.7,
136.6, 145.3, 159.5, 161.2. HRMS (ESI(+)) Calcd for C36H43O4Si
[M+H]+: 567.2925, Found: 567.2928.
1H NMR (400 MHz, CDCl3) d (ppm): 0.90–0.94 (m, 1H), 1.25 (dd,
J = 8.8, 4.8 Hz, 1H), 1.76–1.85 (m, 1H), 2.72–2.78 (m, 1H), 3.41–3.49
(m, 1H), 3.76 (s, 3H), 3.93 (d, J = 10.0 Hz, 1H), 4.04–4.12 (m, 1H),
4.56 (d, J = 10.0 Hz, 1H), 6.42 (t, J = 2.4 Hz, 1H), 6.45–6.49 (m,
1H), 6.50–6.53 (m, 1H), 7.15 (t, J = 8.0 Hz, 1H), 7.19–7.24 (m, 1H),
7.27–7.33 (m, 2H), 7.39–7.43 (m, 2H). 13C NMR (150 MHz, CDCl3)
d (ppm): 17.1, 27.6, 29.5, 55.3, 63.5, 73.4, 101.3, 106.9, 107.2,
126.7, 128.3, 128.7, 130.0, 143.7, 159.4, 160.9. HRMS (ESI(+)) Calcd
for C18H21O3 [M+H]+: 285.1485, Found: 285.1486.
6.1.31. {2-[3-Methoxy-4-(methoxymethyl)phenoxymethyl]-2-
phenylcyclopropyl}methanol (16f)
6.1.27. tert-Butyl({2-[4-methoxy-3-(methoxymethyl)phenoxy-
methyl]-2-phenylcyclopropyl}methoxy)diphenylsilane (15g)
The title compound was synthesized according to the procedure
described for compound 3 from [2-(tert-butyldiphenylsilyloxym-
ethyl)-1-phenylcyclopropyl]methanol 14a and phenol 37
(74.4% yield) as a light yellow oil.
The title compound was synthesized according to the procedure
described for compound 16a from tert-butyl({2-[3-methoxy-4-
(methoxymethyl)phenoxymethyl]-2-phenylcyclopropyl}methoxy)-
diphenylsilane 15f (90.8% yield) as a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 0.93 (t, J = 5.6 Hz, 1H), 1.23–
1.29 (m, 1H), 1.78–1.86 (m, 1H), 2.72–2.78 (m, 1H), 3.35 (s, 3H),
3.42–3.50 (m, 1H), 3.78 (s, 3H), 3.95 (d, J = 10.4 Hz, 1H), 4.05–
4.13 (m, 1H), 4.40 (s, 2H), 4.58 (d, J = 10.4 Hz, 1H), 6.39 (d,
J = 2.4 Hz, 1H), 6.42 (dd, J = 8.0, 2.4 Hz, 1H), 7.18 (d, J = 8.0 Hz,
1H), 7.19–7.24 (m, 1H), 7.28–7.35 (m, 2H), 7.39–7.43 (m, 2H). 13C
NMR (150 MHz, CDCl3) d (ppm): 17.1, 27.6, 29.5, 55.3, 63.5, 73.4,
101.3, 106.9, 107.2, 126.7, 128.3, 128.7, 130.0, 143.7, 159.4,
1H NMR (400 MHz, CDCl3) d (ppm): 0.92 (dd, J = 5.2 Hz, 1H), 1.08
(s, 9H), 1.18 (dd, J = 8.8, 5.2 Hz, 1H), 1.55–1.64 (m, 1H), 3.38 (s, 3H),
3.77 (s, 3H), 3.84 (dd, J = 11.2, 8.0 Hz, 1H), 3.99 (dd, J = 11.2, 6.0 Hz,
1H), 4.14 (d, J = 14.0 Hz, 1H), 4.17 (d, J = 14.0 Hz, 1H), 4.43 (s, 2H),
6.64 (dd, J = 8.8, 3.2 Hz, 1H), 6.72 (d, J = 8.8 Hz, 1H), 6.86 (d,
J = 3.2 Hz, 1H), 7.17–7.44 (m, 11H), 7.65–7.72 (m, 4H). 13C NMR
(150 MHz, CDCl3) d (ppm): 17.2, 20.2, 27.9, 28.4, 30.5, 56.4, 58.8,
64.7, 70.4, 73.1, 100.1, 105.8, 119.7, 127.3, 128.7, 128.7, 129.2,
129.8, 130.6, 131.3, 134.7, 134.7, 136.6, 145.3, 159.5, 161.2. HRMS
(ESI(+)) Calcd for C36H43O4Si [M+H]+: 567.2925, Found: 567.2924.
160.9. HRMS (ESI(+)) Calcd for
374.2326, Found: 374.2327.
C
22H32NO4 [M+C2H5NH3]+:
6.1.32. {2-[4-Methoxy-3-(methoxymethyl)phenoxymethyl]-2-
phenylcyclopropyl}methanol (16g)
The title compound was synthesized according to the procedure
described for compound 16a from tert-butyl({2-[4-methoxy-3-
(methoxymethyl)phenoxymethyl]-2-phenylcyclopropyl}methoxy)-
diphenylsilane 15g (86.7% yield) as a colorless oil.
6.1.28. 2-(Phenoxymethyl)-2-phenylcyclopropyl]methanol
(16c)
The title compound was synthesized according to the procedure
described for compound 16a from tert-butyl({[2-(phenoxymethyl)-
2-phenylcyclopropyl]methoxy})diphenylsilane 16c (87.9% yield) as
a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 0.90 (t, J = 5.2 Hz, 1H), 1.24
(dd, J = 8.8, 5.2 Hz, 1H), 1.75–1.84 (m, 1H), 2.89–2.96 (m, 1H), 3.40–
3.48 (m, 1H), 3.41 (s, 3H), 3.77 (s, 3H), 3.91 (d, J = 9.6 Hz, 1H), 4.05–
4.14 (m, 1H), 4.42 (d, J = 12.8 Hz, 1H), 4.45 (d, J = 12.8 Hz, 1H), 4.53
(d, J = 9.6 Hz, 1H), 6.73–6.75 (m, 2H), 6.92–6.94 (m, 1H), 7.19–7.24
(m, 1H), 7.27–7.33 (m, 2H), 7.39–7.44 (m, 2H). 13C NMR (150 MHz,
CDCl3) d (ppm): 17.1, 27.6, 29.5, 55.3, 63.5, 73.4, 101.3, 106.9,
107.2, 126.7, 128.3, 128.7, 130.0, 143.7, 159.4, 160.9. HRMS
1H NMR (400 MHz, CDCl3) d (ppm): 0.92 (dd, J = 6.0, 4.8 Hz, 1H),
1.26 (dd, J = 8.8, 4.8 Hz, 1H), 1.77–1.86 (m, 1H), 2.80 (dd, J = 7.2,
2.0 Hz, 1H), 3.41–3.48 (m, 1H), 3.94 (d, J = 10.0 Hz, 1H), 4.09
(ddd, J = 16.0, 11.2, 5.2 Hz, 1H), 4.57 (d, J = 10.0 Hz, 1H), 6.85–
6.89 (m, 2H), 6.93–6.98 (m, 1H), 7.19–7.33 (m, 5H), 7.40–7.44
(m, 2H). 13C NMR (150 MHz, CDCl3) d (ppm): 17.2, 20.2, 27.9,
28.4, 30.5, 56.4, 58.8, 64.7, 70.4, 73.1, 100.1, 105.8, 119.7, 127.3,
128.7, 128.7, 129.2, 129.8, 130.6, 131.3, 134.7, 134.7, 136.6,
(ESI(+)) Calcd for
346.2013.
C
20H28NO4 [M+NH4]+: 346.2013, Found:
145.3, 159.5, 161.2. HRMS (ESI(+)) Calcd for
C17H22NO2
[M+NH4]+: 272.1645, Found: 272.1644.
6.1.33. 2-(Phenoxymethyl)-2-phenylcyclopropane-1-carbalde-
hyde (17c)
6.1.29. 2-(4-Methoxyphenoxymethyl)-2-phenylcyclopropyl]
methanol (16d)
The title compound was synthesized according to the procedure
described for compound 16a from tert-butyl({[2-(4-methoxyphen-
The title compound was synthesized according to the procedure
described for compound 17a from 2-(phenoxymethyl)-2-phenyl-
cyclopropyl]methanol 16c (93.4% yield) as a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 1.68 (dd, J = 8.4, 5.2 Hz, 1H),
1.97 (dd, J = 6.0, 5.2 Hz, 1H), 2.40 (ddd, J = 8.4, 6.0, 4.4 Hz, 1H), 4.12
(d, J = 10.0 Hz, 1H), 4.45 (d, J = 10.0 Hz, 1H), 6.78–6.85 (m, 2H),
6.89–6.95 (m, 1H), 7.19–7.37 (m, 5H), 7.42–7.47 (m, 2H), 9.71 (d,
J = 4.4 Hz, 1H).
oxymethyl)-2-phenylcyclopropyl]methoxy})diphenylsilane
(94.6% yield) as a colorless oil.
15d
1H NMR (400 MHz, CDCl3) d (ppm): 0.90 (t, J = 5.2 Hz, 1H), 1.24
(dd, J = 8.8, 5.2 Hz, 1H), 1.76–1.85 (m, 1H), 2.88–2.95 (m, 1H), 3.41–
3.48 (m, 1H), 3.74 (s, 3H), 3.90 (d, J = 10.0 Hz, 1H), 4.05–4.14 (m,
1H), 4.51 (d, J = 10.0 Hz, 1H), 6.80 (s, 4H), 7.19–7.24 (m, 1H),
7.28–7.33 (m, 2H), 7.40–7.44 (m, 2H). 13C NMR (150 MHz, CDCl3)
d (ppm): 17.1, 27.5, 29.7, 55.7, 63.5, 74.5, 114.7, 116.1, 126.6,
128.3, 128.8, 143.8, 152.3, 154.5. HRMS (ESI(+)) Calcd for
6.1.34. 2-(4-Methoxyphenoxymethyl)-2-phenylcyclopropane-1-
carbaldehyde (17d)
The title compound was synthesized according to the proce-
dure described for compound 17a from [2-(3-methoxyphenoxym-
C
18H24NO3 [M+NH4]+: 302.1751, Found: 302.1752.

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6083
ethyl)-2-phenylcyclopropyl]methanol 16d (93.1% yield) as a col-
orless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 1.58 (dd, J = 8.4, 4.8 Hz, 1H),
1.70 (dd, J = 6.0, 4.8 Hz, 1H), 2.13 (dd, J = 8.4, 6.0 Hz, 1H), 3.72 (s,
3H), 4.24 (d, J = 10.0 Hz, 1H), 4.36 (d, J = 10.0 Hz, 1H), 6.75 (s,
4H), 7.23–7.27 (m, 1H), 7.29–7.35 (m, 2H), 7.43–7.46 (m, 2H). 13C
NMR (150 MHz, CDCl3) d (ppm): 19.9, 25.5, 36.0, 55.7, 71.3,
114.5, 116.0, 127.3, 128.4, 128.8, 142.0, 153.2, 154.0, 176.1. HRMS
(ESI(ꢀ)) Calcd for C18H17O4 [MꢀH]ꢀ: 297.1132, Found: 297.1136.
1H NMR (400 MHz, CDCl3) d (ppm): 1.66 (dd, J = 8.0, 5.2 Hz, 1H),
1.95 (dd, J = 6.0, 5.2 Hz, 1H), 2.38 (ddd, J = 8.0, 5.2, 4.4 Hz, 1H), 3.74
(s, 3H), 4.07 (d, J = 10.0 Hz, 1H), 4.40 (d, J = 10.0 Hz, 1H), 6.72–6.80
(m, 4H), 7.23–7.36 (m, 3H), 7.42–7.46 (m, 2H), 9.69 (d, J = 4.4 Hz,
1H).
6.1.40. 2-(3-Methoxyphenoxymethyl)-2-phenylcyclopropane-1-
carboxylic acid (18e)
The title compound was synthesized according to the procedure
described for compound 18a from 2-(3-methoxyphenoxymethyl)-
2-phenylcyclopropane-1-carbaldehyde 17e (89.1% yield) as a white
solid.
6.1.35. 2-(3-Methoxyphenoxymethyl)-2-phenylcyclopropane-1-
carbaldehyde (17e)
The title compound was synthesized according to the procedure
described for compound 17a from [2-(3-methoxyphenoxymethyl)-
2-phenylcyclopropyl]methanol 16e (95.2% yield) as light yellow
oil.
1H NMR (400 MHz, CDCl3) d (ppm): 1.60 (dd, J = 8.0, 5.2 Hz, 1H),
1.72 (dd, J = 6.0, 5.2 Hz, 1H), 2.15 (dd, J = 8.0, 6.0 Hz, 1H), 3.74 (s,
3H), 4.27 (d, J = 10.0 Hz, 1H), 4.40 (d, J = 10.0 Hz, 1H), 6.37–6.48
(m, 2H), 7.10 (t, J = 8.0 Hz, 1H), 7.22–7.28 (m, 2H), 7.29–7.35 (m,
2H), 7.42–7.47 (m, 2H). 13C NMR (150 MHz, CDCl3) d (ppm): 20.0,
25.6, 35.8, 55.2, 70.4, 101.2, 106.7, 106.9, 127.3, 128.4, 128.8,
129.7, 141.9, 160.2, 160.7, 176.3. HRMS (ESI(ꢀ)) Calcd for
1H NMR (400 MHz, CDCl3) d (ppm): 1.68 (dd, J = 8.4, 5.2 Hz, 1H),
1.96 (t, J = 5.2 Hz, 1H), 2.39 (ddd, J = 8.4, 5.2, 4.4 Hz, 1H), 3.75 (s,
3H), 4.11 (d, J = 10.0 Hz, 1H), 4.44 (d, J = 10.0 Hz, 1H), 6.38 (t,
J = 2.4 Hz, 1H), 6.41 (dd, J = 8.0, 2.4 Hz, 1H), 6.48 (dd, J = 8.0,
2.4 Hz, 1H), 7.12 (t, J = 2.4 Hz, 1H), 7.22–7.37 (m, 3H), 7.40–7.46
(m, 2H), 9.70 (d, J = 4.4 Hz, 1H).
C
18H17O4 [MꢀH]ꢀ: 297.1132, Found: 297.1130.
6.1.36. 2-[3-Methoxy-4-(methoxymethyl)phenoxymethyl]-2-
phenylcyclopropane-1-carbaldehyde (17f)
The title compound was synthesized according to the procedure
described for compound 17a from {2-[3-methoxy-4-(methoxy-
6.1.41. 2-[3-Methoxy-4-(methoxymethyl)phenoxymethyl]-2-
phenylcyclopropane-1-carboxylic acid (18f)
The title compound was synthesized according to the procedure
described for compound 18a from 2-[3-methoxy-4-(methoxy-
methyl)phenoxymethyl]-2-phenylcyclopropane-1-carbaldehyde 17f
(64.2% yield) as a white solid.
methyl)phenoxymethyl]-2-phenylcyclopropyl}methanol
(90% yield) as a light yellow oil.
16f
1H NMR (400 MHz, CDCl3) d (ppm): 1.69 (dd, J = 8.0, 4.8 Hz, 1H),
1.97 (dd, J = 5.6, 4.8 Hz, 1H), 2.40 (ddd, J = 8.0, 5.6, 4.0 Hz, 1H), 3.35
(s, 3H), 3.78 (s, 3H), 4.13 (d, J = 10.4 Hz, 1H), 4.39 (s, 2H), 4.47 (d,
J = 10.4 Hz, 1H), 6.34–6.39 (m, 2H), 7.14–7.17 (m, 1H), 7.24–7.30
(m, 1H), 7.31–7.37 (m, 2H), 7.42–7.46 (m, 2H), 9.70 (d, J = 5.2 Hz,
1H).
1H NMR (400 MHz, CDCl3) d (ppm): 1.59 (dd, J = 8.0, 4.8 Hz, 1H),
1.72 (dd, J = 6.0, 4.8 Hz, 1H), 2.15 (dd, J = 8.0, 5.6 Hz, 1H), 3.33 (s,
3H), 3.77 (s, 3H), 4.29 (d, J = 10.0 Hz, 1H), 4.38 (s, 2H), 4.43 (d,
J = 10.4 Hz, 1H), 6.35–6.39 (m, 2H), 7.12–7.16 (m, 1H), 7.23–7.28
(m, 1H), 7.30–7.35 (m, 2H), 7.42–7.46 (m, 2H). 13C NMR (150 MHz,
CDCl3) d (ppm): 20.0, 25.6, 35.8, 55.5, 57.9, 69.3, 70.5, 99.2, 105.1,
119.0, 127.3, 128.4, 128.8, 130.4, 141.9, 158.4, 160.1, 176.0. HRMS
(ESI(ꢀ)) Calcd for C20H21O5 [MꢀH]ꢀ: 341.1394, Found: 341.1393.
6.1.37. 2-[4-Methoxy-3-(methoxymethyl)phenoxymethyl]-2-
phenylcyclopropane-1-carbaldehyde (17g)
The title compound was synthesized according to the procedure
described for compound 17a from {2-[4-methoxy-3-(methoxy-
6.1.42. 2-[4-Methoxy-3-(methoxymethyl)phenoxymethyl]-2-
phenylcyclopropane-1-carboxylic acid (18g)
The title compound was synthesized according to the procedure
described for compound 18a from 2-[4-methoxy-3-(methoxy-
methyl)phenoxymethyl]-2-phenylcyclopropane-1-carbaldehyde 17g
(82.7% yield) as a white solid.
methyl)phenoxymethyl]-2-phenylcyclopropyl}methanol
(92.2% yield) as a light yellow oil.
16g
1H NMR (400 MHz, CDCl3) d (ppm): 1.66 (dd, J = 8.4, 4.8 Hz, 1H),
1.95 (dd, J = 5.6, 4.8 Hz, 1H), 2.34–2.40 (m, 1H), 3.40 (s, 3H), 3.76 (s,
3H), 4.08 (d, J = 10.0 Hz, 1H), 4.42 (d, J = 10.0 Hz, 1H), 4.43 (s, 2H),
6.99 (dd, J = 8.8, 3.2 Hz, 1H), 6.73 (d, J = 8.8 Hz, 1H), 6.88 (d,
J = 3.2 Hz, 1H), 7.24–7.29 (m, 1H), 7.31–7.36 (m, 2H), 7.42–7.46
(m, 2H), 9.68 (d, J = 4.8 Hz, 1H).
1H NMR (400 MHz, CDCl3) d (ppm): 1.55 (dd, J = 8.0, 5.2 Hz, 1H),
1.69 (t, J = 5.2 Hz, 1H), 2.11–2.17 (m, 1H), 3.39 (s, 3H), 3.75 (s, 3H),
4.25 (d, J = 10.0 Hz, 1H), 4.39 (d, J = 10.0 Hz, 1H), 4.42 (s, 2H), 6.67–
6.73 (m, 2H), 6.88–6.90 (m, 1H), 7.22–7.27 (m, 1H), 7.29–7.34 (m,
2H), 7.42–7.46 (m, 2H). 13C NMR (150 MHz, CDCl3) d (ppm): 20.0,
25.6, 35.8, 55.5, 57.9, 69.3, 70.5, 99.2, 105.1, 119.0, 127.3, 128.4,
128.8, 130.4, 141.9, 158.4, 160.1, 176.0. HRMS (ESI(ꢀ)) Calcd for
6.1.38. 2-(Phenoxymethyl)-2-phenylcyclopropane-1-carboxylic
acid (18c)
The title compound was synthesized according to the procedure
described for compound 18a from 2-(phenoxymethyl)-2-phenyl-
cyclopropane-1-carbaldehyde 17c (84.5% yield) as a white solid.
1H NMR (400 MHz, CDCl3) d (ppm): 1.68 (dd, J = 8.4, 5.2 Hz, 1H),
1.97 (dd, J = 6.0, 5.2 Hz, 1H), 2.40 (ddd, J = 8.4, 6.0, 4.4 Hz, 1H), 4.12
(d, J = 10.0 Hz, 1H), 4.45 (d, J = 10.0 Hz, 1H), 6.78–6.85 (m, 2H),
6.89–6.95 (m, 1H), 7.19–7.37 (m, 5H), 7.42–7.47 (m, 2H). 13C
NMR (150 MHz, CDCl3) d (ppm): 20.0, 25.5, 35.9, 70.3, 114.8,
120.8, 127.3, 128.4, 128.9, 129.3, 142.0, 159.0, 176.1. HRMS
(ESI(ꢀ)) Calcd for C17H15O3 [MꢀH]ꢀ: 267.1027, Found: 267.1027.
C
20H21O5 [MꢀH]ꢀ: 341.1394, Found: 341.1393.
6.1.43. 2-(Phenoxymethyl)-2-phenyl-N-(pyridin-2-yl)cyclopropane-
1-carboxamide (19)
The title compound was synthesized according to the procedure
described for compound 4 from 2-(phenoxymethyl)-2-phenylcy-
clopropane-1-carboxylic acid 18c (70.1% yield) as
a white
amorphous solid.
1H NMR (400 MHz, CDCl3) d (ppm): 1.59 (dd, J = 8.0, 5.2 Hz, 1H),
1.90 (t, J = 5.2 Hz, 1H), 2.07–2.13 (m, 1H), 4.32 (d, J = 10.0 Hz, 1H),
4.48 (d, J = 10.0 Hz, 1H), 6.73–6.78 (m, 2H), 6.81–6.86 (m, 1H),
6.95–7.00 (m, 1H), 7.10–7.16 (m, 2H), 7.24–7.29 (m, 1H), 7.30–
7.36 (m, 2H), 7.42–7.48 (m, 2H), 7.59–7.65 (m, 1H), 8.06 (d,
J = 8.4 Hz, 1H), 8.22–8.26 (m, 1H), 8.54 (s, 1H). 13C NMR (150 MHz,
CDCl3) d (ppm): 18.4, 29.7, 35.0, 70.0, 114.0, 114.9, 119.6, 120.7,
6.1.39. 2-(4-Methoxyphenoxymethyl)-2-phenylcyclopropane-1-
carboxylic acid (18d)
The title compound was synthesized according to the procedure
described for compound 18a from 2-(phenoxymethyl)-2-phenyl-
cyclopropane-1-carbaldehyde 17d (92.4% yield) as a white solid.

6084
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
127.2, 128.5, 129.2, 138.3, 142.3, 147.7, 151.4, 158.9, 168.5. HRMS
(ESI(+)) Calcd for C22H21N2O2 [M+H]+: 345.1598, Found: 345.1600.
1H NMR (500 MHz, CDCl3) d (ppm): 1.61 (dd, J = 7.9, 5.3 Hz, 1H),
1.92 (t, J = 5.5 Hz, 1H), 2.12 (t, J = 7.0 Hz, 1H), 3.33 (s, 3H), 3.63 (s,
3H), 4.31 (d, J = 9.4 Hz, 1H), 4.35 (s, 2H), 4.53 (d, J = 9.8 Hz, 1H),
6.27 (d, J = 2.3 Hz, 1H), 6.36 (dd, J = 8.3, 2.3 Hz, 1H), 6.98–7.03 (m,
1H), 7.09 (d, J = 7.9 Hz, 1H), 7.28–7.31 (m, 1H), 7.36 (t, J = 7.6 Hz,
2H), 7.44–7.50 (m, 2H), 7.62–7.67 (m, 1H), 8.08 (d, J = 8.3 Hz,
1H), 8.26 (dd, J = 4.7, 1.0 Hz, 1H), 8.39 (s, 1H).13C NMR (150 MHz,
CDCl3) d (ppm): 18.5, 29.8, 34.9, 55.3, 57.8, 69.3, 70.3, 99.3,
105.2, 114.0, 118.9, 119.6, 127.2, 128.3, 128.5, 130.3, 138.3,
142.2, 147.7, 151.4, 158.3, 160.0, 168.5. HRMS (ESI(+)) Calcd for
6.1.44. 2-(3,4-Dimethoxyphenyl)-2-(phenoxymethyl)-N-
(pyridin-2-yl)cyclopropane-1-carboxamide (20)
To a solution of 2-(3,4-dimethoxyphenyl)-2-(phenoxymethyl)
cyclopropanecarboxylic acid 18b (25 mg, 0.076 mmol) in DMF
(2 ml) was added N,N-diisopropylethyl amine (26.4 ll,
0.152 mmol) followed by HATU (39.1 mg, 0.103 mmol) and 2-
amino pyridine (8.6 mg, 0.091 mmol) at 25 °C. After being stirred
at the same temperature for 10 min. The reaction mixture was
allowed to warm to 60 °C, then stirred for overnight, followed by
concentrated in vacuo. The residue was purified by silica gel col-
umn chromatography using n-heptane/AcOEt (3:1–1:2, v/v) to give
amide 19 (22 mg, 71.4% yield) as a white solid.
C
25H27N2O4 [M+H]+: 419.1965, Found: 419.1966.
6.1.48. 2-[4-Methoxy-3-(methoxymethyl)phenoxymethyl]-2-
phenyl-N-(pyridin-2-yl)cyclopropane-1-carboxamide (24)
The title compound was synthesized according to the procedure
described for compound 19 from 2-[4-methoxy-3-(methoxy-
methyl)phenoxymethyl]-2-phenylcyclopropane-1-carboxylic acid
18g (69.5% yield) as a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 1.58 (dd, J = 8.4, 5.2 Hz, 1H),
1.86 (t, J = 5.2 Hz, 1H), 2.04–2.10 (m, 1H), 3.87 (s, 3H), 3.88 (s, 3H),
4.33 (d, J = 10.0 Hz, 1H), 4.44 (d, J = 10.0 Hz, 1H), 6.74–6.87 (m, 4H),
6.96–7.05 (m, 3H), 7.11–7.18 (m, 2H), 7.60–7.66 (m, 1H), 8.07 (d,
J = 8.4 Hz, 1H), 8.23–8.27 (m, 1H), 8.49 (s, 1H). 13C NMR
(150 MHz, CDCl3) d (ppm): 18.5, 29.7, 35.0, 56.0, 56.0, 70.3,
111.0, 112.2, 114.1, 114.9, 119.6, 120.7, 120.7, 129.2, 134.9,
138.3, 147.7, 148.3, 148.8, 151.4, 158.9, 168.7. HRMS (ESI(+)) Calcd
for C24H25N2O4 [M+H]+: 405.1809, Found: 405.1806.
1H NMR (500 MHz, CDCl3) d (ppm): 1.53–1.61 (m, 1H), 1.87 (t,
J = 5.4 Hz, 1H), 2.06–2.12 (m, 1H), 3.32 (s, 3H), 3.72 (s, 3H), 4.26
(d, J = 9.8 Hz, 1H) 4.30–4.38 (m, 2H) 4.44 (d, J = 9.8 Hz, 1H), 6.63
(m, 2H), 6.80 (s, 1H), 6.96–7.03 (m, 1H), 7.23–7.29 (m, 1H), 7.33
(t, J = 7.6 Hz, 2H), 7.42–7.49 (m, 2H), 7.59–7.66 (m, 1H) 8.08 (d,
J = 8.3 Hz, 1H), 8.25 (dd, J = 4.9, 1.0 Hz, 1H), 8.36 (s, 1H).
13C NMR (150 MHz, CDCl3) d (ppm): 18.3, 29.7, 35.1, 56.0, 58.3,
69.4, 70.9, 111.1, 114.0, 114.4, 116.4, 119.5, 127.1, 127.3, 128.5,
138.3, 142.4, 147.7, 151.4, 151.5, 153.0, 168.6. HRMS (ESI(+)) Calcd
for C25H27N2O4 [M+H]+: 419.1965, Found: 419.1964.
6.1.45. 2-(4-Methoxyphenoxymethyl)-2-phenyl-N-(pyridin-2-yl)
cyclopropane-1-carboxamide (21)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(4-methoxyphenoxymethyl)-2-
phenylcyclopropane-1-carboxylic acid 18d (80% yield) as a white
amorphous solid.
6.1.49. 2-(3,4-Dimethoxyphenoxymethyl)-2-phenyl-N-(pyridin-
2-yl)cyclopropane-1-carboxamide (19)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid 18a and 2-amino pyri-
dine (85.0% yield) as a white amorphous solid.
1H NMR (600 MHz, CDCl3) d (ppm): 1.53–1.62 (m, 1H), 1.88 (t,
J = 5.5 Hz, 1H), 2.10 (t, J = 6.8 Hz, 1H), 3.71 (s, 3H), 4.28 (d,
J = 9.8 Hz, 1H), 4.45 (d, J = 9.8 Hz, 1H), 6.70 (d, J = 3.8 Hz, 4H),
7.01 (dd, J = 7.0, 5.5 Hz, 1H), 7.26–7.30 (m, 1H), 7.35 (t, J = 7.6 Hz,
2H), 7.47 (d, J = 7.9 Hz, 2H), 7.60–7.69 (m, 1H), 8.09 (d, J = 8.3 Hz,
1H), 8.22–8.30 (m, 1H), 8.41 (br s, 1H). 13C NMR (150 MHz, CDCl3)
d (ppm): 18.4, 29.7, 35.1, 55.7, 71.0, 114.0, 114.4, 116.1, 119.6,
127.1, 128.4, 128.5, 138.3, 142.3, 147.7, 151.4, 153.1, 153.9,
168.6. HRMS (ESI(+)) Calcd for C23H23N2O3 [M+H]+: 375.1703,
Found: 375.1705.
1H NMR (500 MHz, MHz, CDCl3) d (ppm): 1.57 (d, J = 5.4 Hz, 1H),
1.89 (t, J = 5.6 Hz, 1H), 2.10 (dd, J = 7.6, 6.1 Hz, 1H), 3.63 (s, 3H),
3.76 (s, 3H), 4.24 (d, J = 9.8 Hz, 1H), 4.47 (d, J = 9.8 Hz, 1H), 6.24–
6.32 (m, 2H), 6.64 (d, J = 8.3 Hz, 1H), 6.94–7.03 (m, 1H), 7.28 (d,
J = 7.3 Hz, 1H) 7.34 (t, J = 7.6 Hz, 2H), 7.43–7.47 (m, 2H), 7.59–
7.70 (m, 1H), 8.07 (d, J = 8.3 Hz, 1H), 8.21–8.29 (m, 1H), 8.46 (s,
1H). 13C NMR (150 MHz, CDCl3) d (ppm): 18.4, 29.8, 35.0, 55.6,
56.4, 71.0, 101.3, 104.6, 111.6, 114.0, 119.6, 127.2, 128.3, 128.5,
138.3, 142.3, 143.5, 147.7, 149.5, 151.4, 153.6, 168.5. HRMS
(ESI(+)) Calcd for C24H25N2O4 [M+H]+: 405.1809.2456, Found:
405.1809.
6.1.46. 2-(3-Methoxyphenoxymethyl)-2-phenyl-N-(pyridin-2-yl)
cyclopropane-1-carboxamide (22)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3-methoxyphenoxymethyl)-2-
phenylcyclopropane-1-carboxylic acid 18e (71.7% yield) as a white
amorphous solid.
1H NMR (500 MHz, CDCl3) d (ppm): 1.59 (dd, J = 8.1, 5.1 Hz, 1H),
1.90 (t, J = 5.4 Hz, 1H), 2.10 (t, J = 6.8 Hz, 1H), 3.64 (s, 3H), 4.30 (d,
J = 9.8 Hz, 1H), 4.49 (d, J = 9.8 Hz, 1H), 6.30 (t, J = 2.4 Hz, 1H), 6.34–
6.43 (m, 2H), 6.96 (t, J = 6.1 Hz, 1H), 7.03 (t, J = 8.1 Hz, 1H), 7.24–
7.28 (m, 1H), 7.33 (t, J = 7.6 Hz, 2H), 7.44 (d, J = 7.8 Hz, 2H), 7.62
(t, J = 7.7 Hz, 1H), 8.07 (d, J = 8.8 Hz, 1H), 8.17–8.27 (1 H, m), 8.62
(br s, 1H). 13C NMR (150 MHz, CDCl3) d (ppm): 18.4, 29.8, 34.9,
55.1, 70.2, 101.2, 106.7, 106.9, 114.0, 119.6, 127.2, 128.4, 128.5,
129.6, 138.3, 142.3, 147.7, 151.4, 160.2, 160.6, 168.5. HRMS
(ESI(+)) C23H23N2O3 [M+H]+: 375.1703, Found: 375.1703.
6.1.50. 2-(3,4-Dimethoxyphenoxymethyl)-2-phenyl-N-(pyridin-
3-yl)cyclopropane-1-carboxamide (26)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid 18a and 3-amino pyri-
dine (60.9% yield) as a colorless oil.
1H NMR (600 MHz, CDCl3) d (ppm) 1.62 (dd, J = 7.7, 5.5 Hz, 1H),
1.91 (t, J = 5.5 Hz, 1H), 2.14 (dd, J = 7.7, 6.2 Hz, 1H), 3.67 (s, 3H),
3.79 (s, 3H), 4.26 (d, J = 9.8 Hz, 1H), 4.54 (d, J = 9.8 Hz, 1H), 6.28–
6.38 (m, 2H), 6.68 (s, 1H), 7.23 (dd, J = 8.3, 4.9 Hz, 1H), 7.30 (d,
J = 7.2 Hz, 1H), 7.37 (t, J = 7.6 Hz, 2H), 7.46 (d, J = 7.6 Hz, 2H),
7.71 (br s, 1H), 8.09 (d, J = 7.6 Hz, 1H), 8.34 (d, J = 4.2 Hz, 1H),
8.54 (br s, 1H). 13C NMR (150 MHz, CDCl3) d (ppm): 18.9, 30.1,
34.4, 55.7, 56.4, 71.0, 101.2, 104.9, 111.8, 123.6, 127.2, 127.2,
127.8, 128.6, 134.8, 141.0, 142.2, 143.7, 145.3, 149.7, 153.3,
168.6. HRMS (ESI(+)) Calcd for C24H25N2O4 [M+H]+: 405.1809,
Found: 405.1810.
6.1.47. 2-[3-Methoxy-4-(methoxymethyl)phenoxymethyl]-2-
phenyl-N-(pyridin-2-yl)cyclopropane-1-carboxamide (23)
The title compound was synthesized according to the procedure
described for compound 19 from 2-[3-methoxy-4-(methoxy-
methyl)phenoxymethyl]-2-phenylcyclopropane-1-carboxylic acid
18f (65.5% yield) as a colorless gum.

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6085
6.1.51. 2-(3,4-Dimethoxyphenoxymethyl)-2-phenyl-N-(pyridin-
6.1.55. 2-{[3,4-Dimethoxyphenoxy]methyl]-2-phenylcyclopro-
4-yl)cyclopropane-1-carboxamide (27)
pyl}-5-fluoro-1H-benzimidazole (31)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid (18a) and 4-amino
pyridine (56.8% yield) as a white amorphous solid.
To a solution of 2-(3,4-dimethoxyphenyl)oxymethyl-2-phenyl-
cyclopropanecarboxylic acid 18a (100 mg, 0.305 mmol) in DCM
(6 ml) was added oxalyl chloride (52.4 ll, 0.61 mmol) and cat.
DMF (1 drop), and the reaction mixture was stirred at room tem-
perature for 1 h. The reaction mixture was concentrated in vacuo
afforded crude acylchloride.
1H NMR (600 MHz, CDCl3) d (ppm): 1.63–1.66 (m, 1H), 1.91 (t,
J = 5.5 Hz, 1H), 2.12 (dd, J = 7.9, 6.0 Hz, 1H), 3.66 (s, 3H), 3.79 (s,
3H), 4.23 (d, J = 9.8 Hz, 1H), 4.55 (d, J = 9.8 Hz, 1H), 6.29–6.33 (m,
2H), 6.68 (d, J = 8.7 Hz, 1H), 7.30 (d, J = 7.2 Hz, 1H), 7.37 (t,
J = 7.5 Hz, 2H), 7.42–7.47 (m, 4H), 7.87 (s, 1H), 8.47 (d, J = 5.5 Hz,
2H). 13C NMR (150 MHz, CDCl3) d (ppm): 19.0, 30.4, 34.7, 55.7,
56.4, 71.0, 101.3, 105.1, 111.8, 113.4, 127.2, 127.8, 128.6, 142.0,
143.8, 144.9, 149.6, 150.7, 153.3, 168.8. HRMS (ESI(+)) Calcd for
To
a solution of 3,4-diamono-1-fluorobenzene (51.9 mg,
0.412 mmol) in THF, crude acylchloride in THF was added. The
reaction mixture was stirred at room temperature for 30 min then
reflux for 1 h. The reaction mixture was concentrated in vacuo. The
residue was dissolved in AcOH (6 ml). The reaction mixture was
stirred at reflux for 2 h. After the reaction mixture was concen-
trated in vacuo, the residue was partitioned between EtOAc and
satd NaHCO3 solution. The organic layer was dried over MgSO4,
concentrated in vacuo. The residue was purified by NH-silica gel
column chromatography using n-heptane/ethyl acetate (9:1–1:2,
v/v) to give benzimidazole 33 (80 mg, 62.7% yield).
C
24H25N2O4 [M+H]+: 405.1809, Found: 405.1808.
6.1.52. N-(5-Methyl-1,3-thiazol-2-yl)-2-[(3,4-dimethoxyphenyl)-
oxymethyl]-2-phenylcyclopropanecarboxamide (28)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid (18a) and 2-amino-4-
methylthiazole (39.9% yield) as a white solid.
1H NMR (400 MHz, CDCl3) d (ppm): 1.73 (dd, J = 8.8, 5.6 Hz, 1H),
2.00 (t, J = 6.0 Hz, 1H), 2.66 (d, J = 8.8, 6.0 Hz, 1H), 3.64 (s, 3H), 3.80
(s, 3H), 3.99 (d, J = 10.0 Hz, 1H), 4.38 (d, J = 10.0 Hz, 1H), 6.12–6.15
(m, 2H), 6.55–6.58 (m, 1H), 6.92–6.99 (m, 1H), 7.28–7.95 (m, 7H),
9.66 (br s, 1H). 13C NMR (150 MHz, CDCl3) d (ppm): 18.2, 23.5, 33.4,
55.7, 56.3, 73.0, 97.1, 101.4, 105.4, 105.7, 110.5, 111.6, 127.1, 128.3,
128.6, 130.1, 139.6, 142.1, 144.0, 149.6, 152.9, 158.7, 160.3. HRMS
1H NMR (600 MHz, CDCl3) d (ppm): 1.73 (dd, J = 7.9, 5.3 Hz, 1H),
1.98 (t, J = 5.7 Hz, 1H), 2.20–2.25 (m, 4H), 3.69 (s, 3H), 3.78 (s, 3H),
4.28 (d, J = 9.8 Hz, 1H), 4.52 (d, J = 9.8 Hz, 1H), 6.31–6.33 (m, 2H),
6.58 (s, 1H), 6.67 (d, J = 8.0 Hz, 1H), 7.29–7.38 (m, 3H), 7.46 (d,
J = 7.5 Hz, 2H), 11.97 (br s, 1H). 13C NMR (150 MHz, CDCl3) d
(ppm): 11.4, 19.0, 29.3, 35.1, 55.6, 56.4, 71.0, 101.5, 105.0, 111.6,
127.0, 127.2, 128.0, 128.5, 133.6, 142.1, 143.7, 149.5, 153.5,
(ESI(+)) Calcd for
419.1765.
C
25H24FN2O3 [M+H]+: 419.1765, Found:
158.2, 167.9. HRMS (ESI(+)) Calcd for
425.1530, Found: 425.1531.
C
23H25N2O4S [M+H]+:
6.1.56. N-(4-Cyanopyridin-2-yl)-2-[(3,4-dimethoxyphenyl)-
oxymethyl]-2-phenylcyclopropanecarboxamide (32)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid 18a and 2-amino-4-
cyanopyridine (11.4% yield) as a white solid.
6.1.53. N-(4-Methyl-1,3-thiazol-2-yl)-2-[(3,4-dimethoxyphenyl)-
oxymethyl]-2-phenylcyclopropanecarboxamide (29)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid 18a and 2-amino-5-
methylthiazole (39.9% yield) as a colorless oil.
1H NMR (600 MHz, CDCl3) d (ppm): 1.64 (dd, J = 7.9, 5.3 Hz, 1H),
1.93 (t, J = 5.5 Hz, 1H), 2.13 (dd, J = 7.9, 6.0 Hz, 1H), 3.70 (s, 3H),
3.79 (s, 3H), 4.26 (d, J = 9.8 Hz, 1H), 4.49 (d, J = 9.8 Hz, 1H), 6.29–
6.31 (m, 2H), 6.68 (d, J = 8.7 Hz, 1H), 7.21 (dd, J = 5.1, 1.3 Hz, 1H),
7.31 (d, J = 7.2 Hz, 1H), 7.38 (t, J = 7.6 Hz, 2H), 7.44–7.50 (m, 2H),
8.36–8.42 (m, 2H), 8.46 (s, 1H). 13C NMR (150 MHz, CDCl3) d
(ppm): 18.8, 29.8, 35.6, 55.7, 56.4, 70.8, 101.2, 104.8, 111.6,
115.9, 116.4, 120.8, 122.3, 127.3, 128.2, 128.6, 141.9, 143.7,
148.9, 149.6, 152.0, 153.3, 168.9. HRMS (ESI(+)) Calcd for
1H NMR (600 MHz, CDCl3) d (ppm): 1.64 (dd, J = 7.9, 4.9 Hz, 1H),
1.94 (t, J = 5.5 Hz, 1H), 2.10 (dd, J = 7.7, 6.2 Hz, 1H), 2.29 (s, 3H),
3.69 (s, 3H), 3.78 (s, 3H), 4.21 (d, J = 9.8 Hz, 1H), 4.47 (d,
J = 9.8 Hz, 1H), 6.30 (m, 2H), 6.47 (s, 1H), 6.65 (d, J = 8.3 Hz, 1H),
7.29 (d, J = 7.6 Hz, 1H), 7.35 (t, J = 7.5 Hz, 2H), 7.43 (d, J = 7.5 Hz,
2H), 9.59 (br s, 1H). 13C NMR (150 MHz, CDCl3) d (ppm): 17.0,
18.8, 28.6, 35.7, 55.6, 56.4, 71.3, 101.5, 104.9, 108.1, 111.6, 127.3,
128.3, 128.6, 141.9, 143.7, 146.9, 149.5, 153.4, 157.6, 167.7. HRMS
C
25H24N3O4 [M+H]+: 430.1761, Found: 430.1761.
(ESI(+)) Calcd for
425.1531.
C
23H25N2O4S [M+H]+: 425.1530, Found:
6.1.57. N-(5-Cyanopyridin-2-yl)-2-[(3,4-dimethoxyphenyl)-
oxymethyl]-2-phenylcyclopropanecarboxamide (33)
To
a solution of trichloromethyl chloroformate (0.113 ml,
0.915 mmol) in 1,4-dioxane (6 ml) was added a solution of carbox-
ylic acid 18a (300 mg, 0.915 mmol) in 1,4-dioxane (4 ml) and TEA
(0.255 ml, 1.83 mmol) dropwise. After the reaction mixture was
stirred at room temperature for 15 min, 2-amino-5-cyanopyridine
(545 mg, 4.58 mmol) in 1,4-dioxane was added. The reaction mix-
ture was stirred at room temperature for overnight. The resulting
mixture was partitioned between EtOAc and satd NaHCO3 solution,
and the organic layer was dried over MgSO4, and concentrated in
vacuo. The residue was purified by NH-silica gel column chroma-
tography using n-heptane/ethyl acetate (19:1–1:1, v/v) to give
amide 32 (226 mg, 57.5% yield).
6.1.54. N-(5-Methyl-1,2-oxazol-3-yl)-2-[(3,4-dimethoxyphenyl)-
oxymethyl]-2-phenylcyclopropane-1-carboxamide (30)
The title compound was synthesized according to the procedure
described for compound 19 from 2-(3,4-dimethoxyphenyl)oxym-
ethyl-2-phenylcyclopropanecarboxylic acid 18a and 3-amino-5-
methylisoxazole (21.6% yield) as a white solid.
1H NMR (600 MHz, CDCl3) d (ppm): 1.58–1.63 (m, 1H), 1.87–
1.92 (m, 1H), 2.17–2.22 (m, 1H), 2.34 (s, 3H), 3.73 (s, 3H), 3.79 (s,
3H), 4.21 (d, J = 9.8 Hz, 1H), 4.47 (d, J = 9.8 Hz, 1H), 6.30 (dd,
J = 8.9, 2.8 Hz, 1H), 6.35 (d, J = 2.6 Hz, 1H), 6.59 (s, 1H), 6.68 (d,
J = 9.1 Hz, 1H), 7.29 (d, J = 7.2 Hz, 1H), 7.35 (t, J = 7.6 Hz, 2H),
7.45–7.51 (m, 2H), 9.17 (br s, 1H). 13C NMR (150 MHz, CDCl3) d
(ppm): 12.6, 18.7, 28.9, 35.3, 55.6, 56.4, 70.9, 96.5, 101.3, 104.5,
111.6, 127.2, 128.5, 142.1, 143.6, 149.6, 153.6, 158.2, 168.3,
169.8. HRMS (ESI(+)) Calcd for C23H25N2O5 [M+H]+: 409.1758,
Found: 409.1759.
1H NMR (400 MHz, CDCl3) d (ppm): 1.62–1.65 (m, 1H), 1.94 (t,
J = 5.6 Hz, 1H), 2.14 (dd, J = 8.0, 6.0 Hz, 1H), 3.67 (s, 3H), 3.77 (s,
3H), 4.22 (d, J = 9.6 Hz, 1H), 4.48 (d, J = 10.4 Hz, 1H), 6.25–6.29
(m, 2H), 6.64 (d, J = 8.4 Hz, 1H), 7.28–7.46 (m, 5H), 7.84 (dd,
J = 8.8, 1.6 Hz, 1H), 8.20 (d, J = 8.4 Hz, 1H), 8.52 (d, J = 1.6 Hz, 1H),

6086
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
8.70 (s, 1H). 13C NMR (150 MHz, CDCl3) d (ppm): 19.0, 29.9, 35.8,
55.7, 56.4, 70.8, 101.0, 104.6, 104.9, 111.7, 113.5, 116.7, 127.4,
128.2, 128.6, 141.5, 141.8, 143.7, 149.6, 151.6, 153.4, 153.7,
169.0. HRMS (ESI(+)) Calcd for C25H24N3O4 [M+H]+: 430.1761,
Found: 430.1761.
1H NMR (600 MHz, CDCl3) d (ppm): 3.43 (s, 3H), 3.81 (s, 3H),
4.50 (s, 2H), 5.05 (s, 2H) 6.78–6.82 (m, 1H), 6.85–6.90 (m, 1H),
7.07 (d, J = 3.0 Hz, 1H), 7.30–7.35 (m, 1H), 7.36–7.47 (m, 4H). 13C
NMR (150 MHz, CDCl3) d (ppm): 56.0, 58.4, 69.4, 70.7, 111.3,
114.3, 115.8, 127.5, 127.8, 127.8, 128.5, 137.4, 151.5, 152.8. HRMS
(ESI(+)) Calcd for
276.1593.
C
16H22NO3 [M+NH4]+: 276.1594, Found:
6.1.58. (+)-N-(5-Cyanopyridin-2-yl)-2-[(3,4-dimethoxyphenyl)-
oxymethyl]-2-phenylcyclopropanecarboxamide (33a) and (ꢀ)-
N-(5-cyanopyridin-2-yl)-2-[(3,4-dimethoxyphenyl)oxymethyl]-
2-phenylcyclopropanecarboxamide (33b)
6.1.61. 4-Methoxy-3-methoxymethyl phenol (37)
To solution of 4-benzyloxy-1-methoxy-2-methoxymethyl
a
The racemic compound 33 was separated by preparative HPLC
on CHIRALPAKÒ AD-H column (2 cm*25 cm); eluent EtOH; flow
rate 1 ml/min to provide (+)-33a: tR = 16.25–20.63 min (94 mg,
23.9% yield), (ꢀ)-33b: tR = 29.59–37.28 min (80 mg, 20.4% yield)
as white amorphous solids.
benzene 36 (5.13 g, 19.9 mmol) in EtOH (60 ml) was added
Pd(OH)2-C (15 wt.%, 697 mg). The reaction mixture was stirred
under a hydrogen atmosphere for 3 h at room temperature. EtOAc
was added to the reaction mixture. Then was filtered through cel-
ite-pad. The filtrate was concentrated in vacuo. The residue was
purified by silica gel column chromatography using n-heptane/
AcOEt (4:1–2:1, v/v) to give phenol 37 (3.2 g, 86.5% yield) as a
white solid. The white solid was washed with n-heptane.
1H NMR (400 MHz, CDCl3) d (ppm): 1.62–1.65 (m, 1H), 1.94 (t,
J = 5.6 Hz, 1H), 2.14 (dd, J = 8.0, 6.0 Hz, 1H), 3.67 (s, 3H), 3.77 (s,
3H), 4.22 (d, J = 9.6 Hz, 1H), 4.48 (d, J = 10.4 Hz, 1H), 6.25–6.29 (m,
2H), 6.64 (d, J = 8.4 Hz, 1H), 7.28–7.46 (m, 5H), 7.84 (dd, J = 8.8,
1.6 Hz, 1H), 8.20 (d, J = 8.4 Hz, 1H), 8.52 (d, J = 1.6 Hz, 1H), 8.70 (s,
1H). 13C NMR (150 MHz, CDCl3) d (ppm): 19.0, 29.9, 35.8, 55.7,
56.4, 70.8, 101.0, 104.6, 104.9, 111.7, 113.5, 116.7, 127.4, 128.2,
128.6, 141.5, 141.8, 143.7, 149.6, 151.5, 153.4, 153.7, 169.0. HRMS
(ESI(+)) Calcd for C25H24N3O4 [M+H]+: 430.1761, Found: 430.1761.
1H NMR (600 MHz, CDCl3) d (ppm): 3.44 (s, 3H), 3.80 (s, 3H),
4.48 (s, 2H), 4.60 (s, 1H), 6.71–6.80 (m, 2H), 6.89 (d, J = 2.6 Hz,
1H). 13C NMR (150 MHz, CDCl3) d (ppm): 56.0, 58.4, 69.2, 111.6,
114.5, 115.9, 127.9, 149.4, 151.3. HRMS (ESI(+)) Calcd for
C
11H20NO3 [M+C2H5NH3]+: 214.1438, Found: 214.1437.
6.1.62. 4-(Benzyloxy)-2-methoxy-1-(methoxymethyl)benzene
(39)
6.1.59. 5-Benzyloxy-2-methoxyphenyl methanol (35)
To a solution of 5-hydroxy-2-methoxybenzaldehyde 34 (4.0 g,
26.3 mmol) in DMF (30 ml) was added K2CO3 (4 g, 28.9 mmol)
and BnBr (3.44 ml, 28.9 mmol). The reaction mixture was stirred
for 19 h at 55 °C. Additional BnBr (3 ml, 25.2 mmol) was added to
the mixture. Then was stirred for 2 h at 55 °C. The reaction mixture
was partitioned with water and EtOAc. The organic layer was
washed with satd NH4Cl, 1 N aq NaOH, water, brine and dried over
MgSO4 and concentrated in vacuo. The residue was used next step
without further purification.
To a suspension of 4-benzyloxy-2-methoxybenzaldehyde 38
(5.46 g, 22.5 mmol) in MeOH (100 ml) was added NaBH4
(850 mg, 22.5 mmol) at 0 °C. The reaction mixture was stirred at
same temperature for 1 h. Water was added to the reaction and
concentrated in vacuo. The residue was partitioned between EtOAc
and water. The organic layer was washed with water, brine and
dried over MgSO4, concentrated in vacuo. The residue was used
for next step without further purification.
To a solution of alcohol mention above in NMP (70 ml) was
added sodium hydride (1.1 g, 27.6 mmol, 60% in oil) at 0 °C. After
stirred for 10 min for same temperature, the reaction mixture
was allowed warm to room temperature and stirred for 30 min.
Iodomethane (1.72 ml, 27.6 mmol) was added to the reaction mix-
ture. Then was stirred for 1.5 h at room temperature. The reaction
mixture was quenched with ice-water and extracted with EtOAc.
The organic layer was washed with satd NH4Cl, brine and dried
over MgSO4, concentrated in vacuo. The residue was purified by sil-
ica gel column chromatography using n-heptane/AcOEt (6:1–3:1,
v/v) to give ether 39 (5.68 g, 95.6% yield) as a colorless oil.
1H NMR (400 MHz, CDCl3) d (ppm): 3.38 (s, 3H), 3.80 (s, 3H),
4.42 (s, 2H), 5.06 (s, 2H), 6.52–6.56 (m, 2H), 7.20–7.24 (m, 1H),
7.30–7.46 (m, 5H). 13C NMR (150 MHz, CDCl3) d (ppm): 55.5,
58.0, 69.4, 70.2, 99.4, 104.9, 119.3, 127.5, 128.0, 128.6, 130.5,
137.0, 158.6, 159.9. HRMS (ESI(+)) Calcd for C16H19O3 [M+H]+:
259.1329, Found: 259.1329.
To a suspension of aldehyde in MeOH (120 ml), NaBH4 (992 mg,
26.3 mmol) was added at 0 °C. The reaction was stirred for 30 min
for same temperature. Then the reaction mixture was stirred for
2 h at room temperature. Water was added to the reaction mixture
and concentrated in vacuo. The residue was partitioned between
water and EtOAc. The organic layer was washed with water, brine,
dried over MgSO4 and concentrated in vacuo. The residue was puri-
fied by silica gel column chromatography using n-heptane/AcOEt
(6:1–2:1, v/v) to give alcohol 35 (6.0 g, 93.4% yield) as a light yellow
solid.
1H NMR (600 MHz, CDCl3) d (ppm): 2.30 (t, J = 6.6 Hz, 1H), 3.84
(s, 3H), 4.67 (d, J = 6.4 Hz, 2H), 5.04 (s, 2H), 6.78–6.83 (m, 1H),
6.85–6.90 (m, 1H), 6.99 (d, J = 3.0 Hz, 1H) 7.30–7.36 (m, 1H),
7.37–7.47 (m, 4H). 13C NMR (150 MHz, CDCl3) d (ppm): 55.8,
62.2, 70.7, 111.1, 114.2, 116.0, 127.5, 127.9, 128.6, 130.1, 137.3,
151.8, 152.8. HRMS (ESI(+)) Calcd for C17H24O3 [M+C2H5NH3]+:
290.1751, Found: 290.1749.
6.1.63. 3-Methoxy-4-methoxymethyl-phenol (40)
6.1.60. 5-Benzyloxy-2-methoxyphenyl methanol (36)
To
a
solution of 4-(benzyloxy)-2-methoxy-1-(methoxy-
To a solution of 5-benzyloxy-2-methoxyphenyl methanol 35 in
NMP (60 ml) was added sodium hydride (984 mg, 24.6 mmol, 60%
in oil) at 0 °C. After stirred for 10 min for same temperature, the
reaction mixture was allowed warm to room temperature and stir-
red for 30 min. Iodomethane (1.91 ml, 30.8 mmol) was added to
the reaction mixture. Then was stirred for 2 h at room tempera-
ture. The reaction mixture was quenched with ice-water and
extracted with EtOAc. The organic layer was washed with satd
NH4Cl, water, brine and dried over MgSO4, concentrated in vacuo.
The residue was purified by silica gel column chromatography
using n-heptane/AcOEt (6:1–3:1, v/v) to give ether 36 (5.13 g,
96.9% yield) as a colorless oil.
methyl)benzene 39 (5.68 g, 22 mmol) in EtOH (70 ml) was added
Pd(OH)2-C (15 wt.%, 772 mg). The reaction mixture was stirred
under a hydrogen atmosphere for 2 h at room temperature. EtOAc
was added to the reaction mixture. Then was filtered through cel-
ite-pad. The filtrate was concentrated in vacuo. The residue was
purified by silica gel column chromatography using n-heptane/
AcOEt (4:1–2:1, v/v) to give phenol 40 (3.2 g, 86.5% yield) as a
white solid. This compound should be stored at low temperature.
1H NMR (400 MHz, CDCl3) d (ppm): 3.39 (s, 3H), 3.75 (s, 3H),
4.43 (s, 2H), 5.60 (s, 1H), 6.34 (dd, J = 8.0,2.4 Hz, 1H), 6.36 (d,
J = 2.4 Hz, 1H), 7.30 (d, J = 2.4 Hz, 1H).

Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
6087
6.2. Biology
of 10–11 weeks, mice were anesthetized with ketamine/xylazine
(120 mg/kg, 10:1, i.p.) and implanted with two electrodes each
(1.1 mm anterior and 4.6 mm posterior from bregma; 1.45 mm
right of the sagittal suture) for recording of brain superficial (dura
mater) brain wave activity (EEG) and intranuchal neck muscle
activity (EMG). After surgery, animals received kanamycin and
were hardwired to the recording equipment, with subsequent
recovery and habituation to recording conditions for 2 weeks.
Two studies were conducted, one with zolpidem to validate
experimental procedure, and one with 33b. Mice were orally dosed
immediately prior to onset of dark period (lights out). Electrical sig-
nals (EEG/EMG) were amplified, filtered (50 Hz line) and recorded
at 128 Hz sampling rate (VitalRecorder software, KisseiComtech)
for later off-line analysis (SleepSign v3 software, KisseiComtech).
Continuous recordings were divided into 10-s epochs and analyzed
by software via an algorithm that determined vigilance state to be
wakefulness (EMG integral indicating movement), non-REM sleep
(delta wave 1–6 Hz activity over 20% of epoch time), and REM sleep
(theta wave 6–11 Hz activity over 35% of epoch time). Automated
analysis results were then verified and if necessary corrected by a
trained observer blinded as to animal treatment.
6.2.1. In vitro binding assays for Ki value evaluation
Human OX1R or OX2R expressing Chinese hamster ovary (CHO)
cells were homogenized in 20 mM HEPES buffer (pH = 7.4) contain-
ing 10 mM MgCl2 and 2 mM EGTA with a Polytron homogenizer
(Kinematica AG, Lucerne, Switzerland). The homogenate was
centrifuged at 1000ꢁg for 10 min at 4 °C and the supernatant
was further centrifuged at 40,000ꢁg for 45 min at 4 °C (Beckman
Coulter, Fullerton, CA). The pellet was resuspended in the same
buffer and stored at ꢀ80 °C before use.
Cell homogenate was diluted to 250 g/mL in 25 mM HEPES buf-
fer (pH = 7.5) containing 5 mM MgCl2, 1 mM CaCl2, 0.5% BSA, 0.1%
sodium azide, and 0.05% TweenÒ 20. Twenty liter of membrane
suspension of OX1R or OX2R containing 5 g protein was mixed
with 10 L of compounds, Orexin A (100 M, Peptide Institute, Osaka,
Japan) solution or vehicle and 10 L of [125I]-Orexin A solution
(2 nM, PerkinElmer, Waltham, MA) and 60 L of assay buffer (final
volume: 100 L) and the mixture incubated for 30 min at rt on a
96 well Flashplate (PerkinElmer). All reaction mixtures were dis-
carded and washed with 200 L of 25 mM HEPES buffer containing
525 mM of NaCl. The radioactivity (disintegrations per minute,
DPM) of each well was measured using a TopCount device (Perkin-
Elmer). The inhibitory activities of test compounds were calculated
using the following formula:
6.2.3.2. Determination of vigilance state time.
Starting at
oral application, 3 h of EEG/EMG recording results were analyzed
and all 10-s epochs judged to be either wakefulness, non-REM
sleep or REM sleep were added respectively to express vigilance
state time in minutes per either 1 h (for time course) or 3 h (for
cumulative analysis).
Inhibition % = 100ꢀ100 ꢁ (T ꢀ N)/(C ꢀ N)
T: DPM in the presence of compounds (test).
N: DPM in the presence of 10 M Orexin A (nonspecific binding).
C: DPM in the absence of compound (control).
All values were determined as averages of 3 wells in a single
measurement.
6.2.3.3. Determination of sleep latency.
Starting at oral
application, time until the first occurrence of 270 s of sleep (with
a maximum of two short 10-s interruptions permitted) was
defined as latency to sleep, expressed in minutes.
6.2.2. In vivo locomotor paradigm evaluation
For measuring the effects of compounds on spontaneous loco-
motor activity, a computerized open field-locomotor activity mea-
suring system was used (VersaMax Animal Activity Monitoring
System, AccuScan Instruments, Inc., Columbus, OH, U.S.A.). This
system automatically measures ambulatory activity in transparent
boxes (210 mm ꢁ 210 mm ꢁ 205 mm) by counting the interrup-
tions of horizontal infrared beam grid spaced 2.5 cm apart in a
frame set at the floor level of the cage. This system furthermore
automatically measures rearing of mice by counting the interrup-
tions of horizontal infrared beams spaced 2.5 cm apart in a frame
set at the 7 cm above the floor.
6.2.3.4. Statistical analysis.
Data are expressed as the
mean SEM. Differences between the vehicle-treated and all com-
pound-treated groups were evaluated using one-way analysis of
variance (ANOVA) followed by Dunnett multiple comparison test.
A value of p < 0.05 (two sided) was considered to indicate statisti-
cal significance. Statistical analyses were performed using the SAS
software package version 8.2 (SAS Institute Japan).
6.3. Physicochemical properties
Male ddY mice (Japan SLC, Inc., Shizuoka, Japan) were kept on a
12-h light/dark cycle with lights off at 19:00.
6.3.1. Aqueous solubility
Solubilities of the test compounds were determined using a
high-throughput HPLC method.35 A 10 mM solution of test com-
pound in DMSO was prepared and diluted to give a 1:100 DMSO
solution of test medium in the microwell of the filter plate. After
gyratory shaking of the filter plate for 15 min at room tempera-
ture, the solution was filtered. An aliquot of this filtered solution
was injected into the HPLC system, which was equipped with an
ODS column and UV detector. Mobile phase A (0.1% TFA, 1%
MeCN in water) and mobile phase B (50:50 MeCN/EtOH, v/v)
were pumped using a linear gradient program. The solubility of
the compounds was determined by comparison to an external
standard. The standard solution was prepared by diluting a
10 mM DMSO solution of test compound with DMSO to yield a
1:100 test compound solution/DMSO mixture in the well of the
microwell plate.
Compound was dissolved in 100% DMSO and then diluted twen-
tyfold with 10% Cremophor EL in saline. One hour after the treat-
ment with vehicle or compound, mice were placed into the open
field and interruptions of infrared beams were counted for one hour.
Locomotor activity was defined as the sum of ambulatory activity
count, rearing activity count and stereotypical behavior count
(repetitive interruptions of infrared beams without ambulation).
Data were expressed as the mean SEM (vehicle group: n = 16,
compound-treated groups: n = 8). The difference between vehicle
control group and compound-treated groups was performed via
one-way analysis of variance (ANOVA) followed by the Dunnett
multiple comparison test. A value of p < 0.05 (two sided) was con-
sidered to be statistically significant.
6.2.3. In vivo sleep experiment evaluation
6.2.3.1. Recordings of wake/sleep behavior.
From the age of
6.4. DMPK study
eight weeks on and throughout the studies, male mice (C57BL/
6NCrlCrlj; Charles River Laboratories Japan) were kept in a noise-
attenuated environment on a 12-h light/12-h dark cycle at 24 °C
and 50% humidity with free access to food and water. At the age
6.4.1. P-gp transport assay
LLC-PK1 and MDR1 gene-transfected LLC-PK1 (LLC-MDR1) cells
were individually seeded at a density of 4.0 ꢁ 105 cells/cm2 onto

6088
Y. Yoshida et al. / Bioorg. Med. Chem. 22 (2014) 6071–6088
porous membrane filters of 24-well cell culture insert plates. Cells
were cultured in 5% CO2 at 37 °C for 5 days to prepare cell mono-
layer, and were used for the transport studies. For transcellular
transport experiments, each cell monolayer was pre-incubated at
37 °C for 2 h in Hanks’ balanced salt solution containing 10 mM
HEPES (HBSS buffer). Transcellular transport experiments were ini-
tiated by adding the HBSS buffer containing test compounds to the
apical or basal side of the cell culture inserts. After incubation at
37 °C for 2 h, a portion of HBSS buffer was sampled from the
opposite compartment of that spiked with test compounds, and
permeated amount of the test compound was analyzed by LC/
MS. The apparent permeability coefficient (Papp) of the test com-
pounds were estimated using Eq. 1, where Q, t, C0, and A represent
permeated amount of test compounds, incubation time, initial con-
centration of test compounds, and membrane area, respectively.
References and notes
1. Sakurai, T.; Amemiya, A.; Ishii, M.; Matsuzaki, I.; Chemelli, R. M.; Tanaka, H.;
Williams, S. C.; Richardson, J. A.; Kozlowski, G. P.; Wilson, S.; Arch, J. R. S.;
Buckingham, R. E. C.; Haynes, A. C.; Carr, S. A.; Annan, R. S.; McNulty, D. E.; Liu,
W. S.; Terrett, J. A.; Elshourbagy, N. A.; Bergsma, D. J.; Yanagisawa, M. Cell 1998,
92, 573.
2. De Lecea, L.; Kilduff, T. S.; Peyron, C.; Gao, X.-B.; Foye, P. E.; Danielson, P. E.;
Fukuhara, C.; Battenberg, E. L. F.; Gautvik, V. T.; Bartlett, F. S., II; Frankel, W. N.;
Van Den Pol, A. N.; Bloom, F. E.; Gautvik, K. M.; Sutcliffe, J. G. Proc. Natl. Acad. Sci.
U.S.A. 1998, 95, 322.
3. Van den Pol, A. N.; Gao, X.-B.; Obrietan, K.; Kilduff, T. S.; Belousov, A. B. J.
Neurosci. 1998, 18, 7962.
4. Beuckmann, C. T.; Yanagisawa, M. J. Mol. Med. (Berl.) 2002, 80, 329.
5. Brisbare-Roch, C.; Dingemanse, J.; Koberstein, R.; Hoever, P.; Aissaoui, H.;
Flores, S.; Mueller, C.; Nayler, O.; van Gerven, J.; de Haas, S. L.; Hess, P.; Qiu, C.;
Buchmann, S.; Scherz, M.; Weller, T.; Fischli, W.; Clozel, M.; Jenck, F. Nat. Med.
2007, 13, 150.
6. Chemelli, R. M.; Willie, J. T.; Sinton, C. M.; Elmquist, J. K.; Scammell, T.; Lee, C.;
Richardson, J. A.; Williams, S. C.; Xiong, Y.; Kisanuki, Y.; Fitch, T. E.; Nakazato,
M.; Hammer, R. E.; Saper, C. B.; Yanagisawa, M. Cell 1999, 98, 437.
7. Willie, J. T.; Chemelli, R. M.; Sinton, C. M.; Tokita, S.; Williams, S. C.; Kisanuki, Y.
Y.; Marcus, J. N.; Lee, C.; Elmquist, J. K.; Kohlmeier, K. A.; Leonard, C. S.;
Richardson, J. A.; Hammer, R. E.; Yanagisawa, M. Neuron 2003, 38, 715.
8. Thannickal, T. C.; Moore, R. Y.; Nienhuis, R.; Ramanathan, L.; Gulyani, S.;
Aldrich, M.; Cornford, M.; Siegel, J. M. Neuron 2000, 27, 469.
Papp ¼ Q=t=C0=A
ð1Þ
Flux ratios (FRs) across the cell monolayer were defined by Eq.
2, where Papp b to a and Papp, a to b represent the Papp in the basal-to-
,
apical direction and the apical-to-basal direction, respectively, and
the corrected FR was defined by Eq. 3.
9. Peyron, C.; Faraco, J.; Rogers, W.; Ripley, B.; Overeem, S.; Charnay, Y.;
Nevsimalova, S.; Aldrich, M.; Reynolds, D.; Albin, R.; Li, R.; Hungs, M.;
Pedrazzoli, M.; Padigaru, M.; Kucherlapati, M.; Fan, J.; Maki, R.; Lammers, G.
J.; Bouras, C.; Kucherlapati, R.; Nishino, S.; Mignot, E. Nat. Med. 2000, 6, 991.
10. Nishino, S. Handb. Clin. Neurol. 2011, 99, 765.
FR ¼ Papp; b to a=Papp; a to b
ð2Þ
Corrected FR ¼ ðFR in LLC-MDR1 cellsÞ=ðFR in LLC-PK1 cellsÞ
11. Akanmu, M. A.; Honda, K. Brain Res. 2005, 1048, 138.
12. Dugovic, C.; Shelton, J. E.; Aluisio, L. E.; Fraser, I. C.; Jiang, X.; Sutton, S. W.;
Bonaventure, P.; Yun, S.; Li, X.; Lord, B.; Dvorak, C. A.; Carruthers, N. I.;
Lovenberg, T. W. J. Pharmacol. Exp. Ther. 2009, 330, 142.
ð3Þ
13. Kummangal, B. A.; Kumar, D.; Mallick, H. N. Behav. Brain Res. 2013, 237, 59.
14. Steiner, M. A.; Gatfield, J.; Brisbane-Roch, C.; Dietrich, H.; Treiber, A.; Jenck, F.;
Boss, C. ChemMedChem 2013, 8, 898.
15. Coleman, P. J.; Renger, J. J. Expert Opin. Ther. Pat. 2010, 20, 307.
16. Bingham, M. J.; Cai, J.; Deehan, M. R. Curr. Opin. Drug Discov. Dev. 2006, 9, 551.
17. Sakurai, T. Nat. Rev. Neurosci. 2007, 8, 171.
18. Etori, K.; Saito, C. Y.; Tsujino, N.; Sakurai, T. Front. Neurosci. 2014, 8, 8.
19. Boss, C.; Brisbare-Roch, C.; Jenck, F. J. Med. Chem. 2009, 52, 891.
20. Roecker, A. J.; Coleman, P. J. Curr. Top. Med. Chem. 2008, 8, 977.
21. Gatfield, J.; Brisbare-Roch, C.; Jenck, F.; Boss, C. Chem. Med. Chem. 2010, 5, 1197.
22. Stasi, L. P.; Artusi, R.; Bovino, C.; Buzzi, B.; Canciani, L.; Caselli, G.; Colace, F.;
Garofalo, P.; Giambuzzi, S.; Larger, P.; Letari, O.; Mandelli, S.; Perugini, L.; Pucci,
S.; Salvi, M.; Toro, P. Bioorg. Med. Chem. Lett. 2013, 23, 2653.
23. Fujimoto, T.; Kunitomo, J.; Tomata, Y.; Nishiyama, K.; Nakashima, M.; Hirozane,
M.; Yoshikubo, S.; Hirai, K.; Marui, S. Bioorg. Med. Chem. Lett. 2011, 21, 6414.
24. Betschart, C.; Hintermann, S.; Behnke, D.; Cotesta, S.; Fendt, M.; Gee, C. E.;
Jacobson, L. H.; Laue, G.; Ofner, S.; Chaudhari, V.; Badiger, S.; Pandit, C.;
Wagner, J.; Hoyer, D. J. Med. Chem. 2013, 56, 7590.
25. Hoever, P.; de Haas, S.; Winkler, J.; Schoemaker, R. C.; Chiossi, E.; van Gerven, J.;
Dingemanse, J. Clin. Pharmacol. Ther. (N.Y., NY, U.S.) 2010, 87, 593.
26. Bettica, P.; Nucci, G.; Pyke, C.; Squassante, L.; Zamuner, S.; Ratti, E.; Gomeni, R.;
Alexander, R. J. Psychopharmacol. 2012, 26, 1058.
27. Winrow, C. J.; Gotter, A. L.; Cox, C. D.; Doran, S. M.; Tannenbaum, P. L.; Breslin,
M. J.; Garson, S. L.; Fox, S. V.; Harrell, C. M.; Stevens, J.; Reiss, D. R.; Cui, D.;
Coleman, P. J.; Renger, J. J. J. Neurogenet. 2011, 25, 52.
28. Cox, C. D.; Breslin, M. J.; Whitman, D. B.; Schreier, J. D.; McGaughey, G. B.;
Bogusky, M. J.; Roecker, A. J.; Mercer, S. P.; Bednar, R. A.; Lemaire, W.; Bruno, J.
G.; Reiss, D. R.; Harrell, C. M.; Murphy, K. L.; Garson, S. L.; Doran, S. M.;
Prueksaritanont, T.; Anderson, W. B.; Tang, C.; Roller, S.; Cabalu, T. D.; Cui, D.;
Hartman, G. D.; Young, S. D.; Koblan, K. S.; Winrow, C. J.; Renger, J. J.; Coleman,
P. J. J. Med. Chem. 2010, 53, 5320.
6.4.2. Determination of plasma and CSF concentrations in mice
Compound 33b was orally administered to male C57BL/6N mice
at a dose of 100 mg/kg. Compound 33b was formulated as a sus-
pension in 0.5% aqueous methylcellulose and saline/Cremophor
EL/DMSO (85:10:5, v/v/v), respectively. At 1 and 3 h after dosing,
CSF was collected from cisterna magna under the anesthesia with
sodium pentobarbital. Immediately after CSF sampling, blood was
collected from the abdominal aorta, and plasma was obtained by
centrifugation. The plasma and CSF samples were stored below
ꢀ20 °C until analysis by LC/MS/MS.
Plasma samples were precipitated with 25 volumes of methanol
including internal standard (10 ng/mL propranolol). Following vor-
tex mixing and centrifugation, the supernatant was filtered, and
the resulting filtrate was injected into the LC/MS/MS system. CSF
samples were mixed with 9 volumes of acetonitrile and 2 volumes
of methanol including internal standard. After vortex mixing, the
mixture was injected into the LC/MS/MS system. Detection was
accomplished by multiple reaction monitoring in positive ioniza-
tion mode. The ratio of the peak area responses relative to the
internal standard were used to construct a standard curve using
linear least squares regression with a 1/x2 weighting. Plasma and
CSF concentrations were reported as the mean SEM.
Acknowledgments
29. Shuto, S.; Ono, S.; Hase, Y.; Kamiyama, N.; Takada, H.; Yamashita, K.; Matsuda,
A. J. Org. Chem 1996, 61, 915.
The authors would like to thank the colleagues who helped gen-
erate data reported in this manuscript. In particular, special thanks
to Eri Ena, Satoko Sasaki, Masaki Kato, and Yumi Yokoyama for
NMR and HRMS analyses, and to Misako Watanabe and Kazuya
Nagaoka for chemoinformatics support.
30. The absolute configuration of 33b has been presumed as Figure 3 by
comparison of chiral HPLC analysis between optically resolved 33b and
enantioselective synthesis of 33b via known intermediate by using (R)-(ꢀ)-
epichlorohydrin.29
31. Pack, A. I.; Galante, R. J.; Maislin, G.; Cater, J.; Metaxas, D.; Lu, S.; Zhang, L.; Von
Smith, R.; Kay, T.; Lian, J.; Svenson, K.; Peters, L. L. Physiol. Genom. 2007, 28, 232.
32. Beuckmann, C. T.; Sinton, C. M.; Williams, S. C.; Richardson, J. A.; Hammer, R.
E.; Sakurai, T.; Yanagisawa, M. J. Neurosci. 2004, 24, 4469.
33. Depoortere, H.; Zivkovic, B.; Lloyd, K. G.; Sanger, D. J.; Perrault, G.; Langer, S. Z.;
Bartholini, G. J. Pharmacol. Exp. Ther. 1986, 237, 649.
Supplementary data
34. Winsky-Sommerer, R. Eur. J. Neurosci. 2009, 29, 1779.
35. Sugaya, T.; Yoshiba, T.; Kajima, T.; Ishihama, Y. Yakugaku Zasshi 2002, 122, 237.
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2014.08.034.