Separation of α-glucosidase-inhibitory and liver X receptor-antagonistic activities of phenethylphenyl phthalimide analogs and generation of LXRα-selective antagonists

Article abstract of DOI:10.1016/j.bmc.2009.05.066

Liver X receptor (LXR) α/β dual agonists are candidate medicaments for the treatment of metabolic syndrome, because their biological actions include increasing cholesterol efflux mediated by LXRβ. However, their clinical application is currently limited by their enhancing effect on triglyceride (TG) synthesis mediated by LXRα. Combination of an LXRα-selective antagonist with an LXRα/β dual agonist may overcome this disadvantage. In the present work, structural development studies of phenethylphenyl phthalimide 9, which possesses LXRα/β dual-antagonistic activity and α-glucosidase-inhibitory activity, led to the LXRα-selective antagonist 23f. Specific α-glucosidase inhibitors were also obtained.

Full text of DOI:10.1016/j.bmc.2009.05.066

Bioorganic & Medicinal Chemistry 17 (2009) 5001–5014
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Separation of
a-glucosidase-inhibitory and liver X receptor-antagonistic
activities of phenethylphenyl phthalimide analogs and generation
of LXR -selective antagonists
a
*
Kazunori Motoshima, Tomomi Noguchi-Yachide, Kazuyuki Sugita, Yuichi Hashimoto, Minoru Ishikawa
Institute of Molecular & Cellular Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
Liver X receptor (LXR)
a
/b dual agonists are candidate medicaments for the treatment of metabolic syn-
Received 28 April 2009
Revised 22 May 2009
Accepted 27 May 2009
Available online 2 June 2009
drome, because their biological actions include increasing cholesterol efflux mediated by LXRb. However,
their clinical application is currently limited by their enhancing effect on triglyceride (TG) synthesis med-
iated by LXR
this disadvantage. In the present work, structural development studies of phenethylphenyl phthalimide
9, which possesses LXR /b dual-antagonistic activity and -glucosidase-inhibitory activity, led to the
LXR -selective antagonist 23f. Specific -glucosidase inhibitors were also obtained.
Ó 2009 Elsevier Ltd. All rights reserved.
a. Combination of an LXRa-selective antagonist with an LXRa/b dual agonist may overcome
a
a
Keywords:
LXR antagonist
a
a
a-Glucosidase inhibitor
Thalidomide analog
1. Introduction
Currently available typical synthetic LXR agonists T0901317
(1)15 and GW3965 (2)16 (Fig. 1) non-selectively activate both LXR
a
Liver X receptors (LXR
a
and LXRb) are members of the nuclear
a is most highly expressed in liver and
and LXRb. They also activate triglyceride (TG) synthesis in the liver
by the up-regulation of sterol regulatory element binding protein
1c (SREBP1c) and fatty acid synthase (FAS), and this activity limits
the clinical utility of these LXR agonists.15,17 There is increasing
evidence that the undesirable stimulation of lipogenesis by LXR li-
receptor superfamily.1 LXR
is abundant in other tissues involved in lipid metabolism. LXRb
has a more widespread pattern of expression, being almost ubiqui-
tous. Their physiological ligands are considered to be oxysterols,
including 24(S),25-epoxycholesterol and 22(R)-hydroxycholes-
terol. Upon ligand binding, LXRs form heterodimers with the reti-
noid X receptor (RXR). The LXR/RXR heterodimer binds to LXR
response elements in promoter regions of specific genes,2,3 modu-
lating gene transcription by recruiting coactivators.4,5 LXRs are
considered to function as cholesterol sensors, and to regulate the
expression of genes associated with cholesterol efflux, absorption
and transport.6,7 The activation of LXRs leads to an increase in plas-
ma HDL levels and net cholesterol efflux via up-regulation of gene
expression of ATP-binding cassette (ABC) membrane transporters,
including ABCA1,8,9 ABCG5, ABCG810 and ABCG1.11,12 ABCA1 func-
tions in transporting cholesterol and phospholipids to ApoA-I,
which is a critical step in cholesterol efflux via HDL. In animal stud-
ies, the activation of LXR led to an increase of HDL level and a de-
crease of atherosclerotic lesions through the induction of
peripheral cholesterol efflux.9,13 There is increasing evidence sug-
gesting that the therapeutic effects of LXR activation on atheroscle-
rosis are associated with stimulation of cholesterol efflux, rather
than simply an increase of serum HDL.13,14
gands is largely attributable to LXR
a
. It has been demonstrated
-null mice
that administration of an LXR /b dual agonist to LXR
a
a
results in increased HDL levels without significant hepatic TG accu-
mulation.18 In these genetically modified mice, hepatic mRNA lev-
els of many lipogenic genes, including SREBP1c, FAS, and
lipoprotein lipase genes, were markedly reduced by LXR agonists,
as compared to wild-type mice. More importantly, LXRa-deficient
macrophages from LXRa-null mice retained the ability to increase
ABCA1,9,19 implying that LXRb is capable of inducing cholesterol ef-
flux in macrophages without inducing significant hepatic fatty acid
synthesis.
These considerations led researchers to focus on LXRb-selective
ligands. LXRa and LXRb are highly related and share 77% amino
acid sequence identity in both the DNA-binding and ligand-binding
domains (LBD).1 Therefore, the design of LXR subtype-selective li-
gands is challenging. To date, only one series of ligands, including
3, appear to be LXRb-selective agonists based on binding assay,
although they act as LXR
As an alternative approach, it seems plausible to speculate that
usage of an LXR -selective antagonist in conjunction with an LXR
/b dual agonist may circumvent the undesirable effect of
increased lipogenesis mediated by LXR while maintaining the
a
/b dual agonists in reporter gene assay.20
a
a
* Corresponding author. Tel.: +81 3 5841 7849; fax: +81 3 5841 8495.
E-mail address: m-ishikawa@iam.u-tokyo.ac.jp (M. Ishikawa).
a
0968-0896/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2009.05.066

5002
K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
Figure 1. Chemical structures of LXR ligands.
enhanced cholesterol efflux mediated by LXRb. There is some
evidence in support of this idea. Herbal extracts, which have
LXRa-selective antagonistic activity, have been reported to sup-
spatially complementary to one fold structure might serve as a
multi-template for structural development of ligands that would
interact specifically with more than 50 different human proteins.
In other words, the structures of ligands that bind to a protein hav-
ing a certain fold structure may be useful for the development of
novel ligands for other proteins possessing the same fold structure.
One of the multi-templates that we have adopted is thalidomide,
which is a drug first launched as a sedative/hypnotic agent, but
withdrawn from the market because of its severe teratogenicity.
Focusing on the potential of the thalidomide template for the treat-
ment of a wide range of diseases, including cancers, diabetes, and
rheumatoid arthritis, we have created compounds thalidomide
analogs with a range of biological activities.30–33 During our stud-
ies, we found that several thalidomide-related phthalimide deriva-
tives, including PP2P (9) (Fig. 1), possess LXR-antagonistic activity
press the expression of LXRa-responsive genes, such as FAS and
SREBP1c.21 These extracts also significantly reduce lipogenesis
and adipocyte differentiation. However, the active ingredients
have not been identified. As another example, an LXR
a/b dual
antagonist, fenofibrate (4), was reported to repress LXR agonist-in-
duced transcription of hepatic lipogenic genes.22 Surprisingly,
however, fenofibrate (4) did not repress LXR-induced transcription
of ABCA1 in liver or in macrophages. Concerning LXR antagonists,
those reported so far include riccardin C (5)23 (LXR
a
partial ago-
antagonist), riccardin
/b dual antagonists and an antagonist with slight
selectivity (7)], and 22(S)-hydroxycholesterol (8)25 (LXR
/b
dual antagonists) (Fig. 1).
nist/LXRb antagonist), riccardin F (6)23 (LXR
C analogs24 [LXR
LXR
a
a
a
a
as well as
a
-glucosidase-inhibitory activity.24,34,35 The co-existence
-glucosidase-inhibitory activity
We have focused on the creation of LXR antagonists using a
multi-template approach based on thalidomide. The multi-tem-
plate approach is based on the reports that the number of three-
dimensional spatial structures (fold structures) of human proteins
is much smaller (more than 50 times smaller) than the number of
human proteins (50,000–70,000).26–29 Therefore, ignoring physi-
cal/chemical interactions, a template/scaffold structure which is
of LXR-modulating activity and
a
may be general, because typical LXR ligands, including T0901317
(1), GW3965 (2), 22-(R)-hydroxycholesterol and riccardin C (5)
were found to possess potent
These results suggested that our previously reported thalido-
mide-related
-glucosidase inhibitors24,34–38 might represent a
superior scaffold structure for the development of LXR antagonists.
a
-glucosidase-inhibitory activity.24
a

K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
5003
The issues to be addressed were therefore the separation of
cosidase-inhibitory and LXR-antagonistic activities and the crea-
tion of LXR -selective antagonists. Here, we describe the design,
synthesis, and structure–activity relationship studies of phenethyl-
a
-glu-
binding site of PP2P (9)/riccardin C (5) on
structure of
(9)/riccardin C (5) to LXR are all unknown, we expected that
a
-glucosidase, the crystal
a
-glucosidase, and the precise binding mode of PP2P
a
PP2P (9) and riccardin C (5) would bind to the same, or at least sim-
phenyl phthalimide derivatives to generate an LXR
antagonist.
a
-selective
ilar, sites in both LXR and a-glucosidase. If this were the case, the
common structure of PP2P (9) and riccardin C (5), represented by
bold lines/letters in Figure 2 might be critical. This speculation,
as well as the results of our previous structural development stud-
ies of riccardin C (5), which afforded novel LXR antagonists lacking
2. Results and discussion
a
-glucosidase-inhibitory activity,24 prompted us to prepare regioi-
2.1. Choice of the phenethylphthalimide skeleton as a scaffold
somers of the phenethyl group and cinnamyl analogs of PP2P (9),
focusing on differences in the position or rigidity of the phenethyl
moiety in PP2P (9) and riccardin C (5). In addition, based on the
structure of riccardins and the critical role of hydroxyl and/or
methoxy group(s) for their biological activities,24 substituent
Initially, we selected our previously reported PP2P (9) as a pro-
totype.24,34,35 We also noted the similarity of the biological activi-
ties (i.e., LXR-antagonistic activity and
a-glucosidase-inhibitory
activity) elicited by PP2P (9) and riccardin C (5).24 Although the
Figure 2. Common structure of riccardin C (5) and PP2P (9), and molecular design of phenethylphenyl phthalimide.
Scheme 1. Synthesis of phenethylphenyl phthalimide analogs. Reagents and conditions: (a) K2CO3, 18-crown-6, CH2Cl2, reflux; (b) H2 10% Pd/C, EtOAc, rt; (c) SnCl2ꢀ2H2O,
EtOAc, reflux; (d) phthalic anhydride or tetrachlorophthalic anhydride, neat, 200 °C; (e) BBr3, CH2Cl2, 0 °C.

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K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
effects involving one or two hydroxyl or methoxy group(s) were
investigated. We also aimed to generate specific -glucosidase
experimental conditions used. Broadly speaking, 40-phenethyl
derivatives are all inactive toward -glucosidase regardless of the
a
a
inhibitors lacking LXRs-antagonistic activity, in order to extend
the multi-template approach. Thus, tetrachlorophthalimide ana-
logs were also designed based on our previous structure–activity
relationship studies indicating that tetrachlorophthalimide analogs
presence of one (Table 2) or two (Table 1) methoxy or hydroxyl
group(s).
Next, based on our previous finding that tetrachlorophthalimide
derivatives tend to possess a-glucosidase-inhibitory activity, tetra-
possess more potent
a
-glucosidase-inhibitory activity than the
chlorophthalimide analogs 24–30 were prepared and their activi-
ties were evaluated (Table 3). Although tetrachlorination of PP2P
(9), that is, compound 24a, resulted in disappearance of the activ-
ity, this seemed to be exceptional. Concerning other derivatives, al-
most all of the tetrachlorophthalimide derivatives showed more
corresponding phthalimide analogs.36–38
Phenylphthalimide analogs were synthesized as shown in
Scheme 1. Briefly, diphenylethene derivatives 11–13 were pre-
pared as E/Z mixtures by Wittig reaction of nitrobenzaldehyde
with ylide 10. For the synthesis of cinnamyl analogs, E derivatives
(12E, 13E) were separated by the use of silica gel column chroma-
tography. After reduction of the nitro group and/or olefin moiety of
compounds 11–13, phenylphthalimide series 19–30 were obtained
by condensation with phthalic anhydride.
potent a-glucosidase-inhibitory activity (Table 3) than that of the
corresponding phthalimide derivatives (Tables 1 and 2), that is,
the orders of the potency of the activity are: 24c > 19c,
25b > 20b, 25c > 20c, 26a > 21a, 26b > 21b, 28a > 22a, 28b > 22b,
30d > 23d, 30e > 23e, and 30g > 23g. Concerning the substituent
effects of methoxy and hydroxyl groups, the case observed in 20-
phenethyl isomers seem exceptional, as mentioned above. In the
case of 30-phenethyl [20a–c (Table 1) and 25a–c (Table 3)] and
2.2. Structure–activity relationship for
activity
a-glucosidase-inhibitory
30-cinnamyl derivatives [27a–c (Table 3)], the potency of
sidase-inhibitory activity decreased in the order of: dihydr-
oxyl > dimethoxy > non-substituted derivatives [20c > 20b
a-gluco-
Antagonistic activities for LXRs were evaluated using a reporter
gene assay method with CMX-GAL4 N-hLXR as the recombinant
receptor gene, TK-MH100x4-LUC as the reporter gene, and the
CMX b-galactosidase gene for normalization, as previously re-
ported.39–41 All compounds did not reduce b-galactosidase activity
at evaluated concentrations unless otherwise noted in Tables 1–3.
None of the compounds tested show agonistic activity under the
experimental conditions used (data not shown).
First, the activities of PP2P (9) were compared with those of its
regio-isomers, 20a and 21a (Table 1). Positional shift of the phen-
ethyl moiety of PP2P (9) from the 20-position to the 30- or 40-posi-
tion resulted in disappearance of a-glucosidase-inhibitory activity.
Introduction of two methoxy or hydroxyl groups into the terminal
(inactive) = 20a (inactive), 25c > 25b > 25a (inactive), and 27c >
27b > 27a]. Concerning the substituent effect elicited by hydroxyl
groups, the 300-position seems to be critical, because 29e showed
moderate activity, while its regio-isomer 29g was inactive
(Table 3). On the other hand, in the case of 40-phenethyl [26a–c
(Table 3)] and 40-cinnamyl derivatives [22a–c (Table 1) and 28a–
c (Table 3)], the order of the potency seems just reversed, that is,
non-substituted > dimethoxy > dihydroxyl derivatives [22a > 22b
(inactive) = 22c (inactive), 26a ꢁ 26b > 26c (inactive), 28a > 28b
> 28c (inactive)]. Concerning the substituent effect elicited by
methoxy groups, the 300-position seems to be critical, as is the case
for hydroxyl groups (vide supra), because 30d showed moderate
activity while its regio-isomer 30f was inactive (Table 3).
phenyl moiety of PP2P (9), that is, compounds 19b and 19c, also re-
sulted in disappearance of a-glucosidase-inhibitory activity (Table
1). However, introduction of two hydroxyl groups into the corre-
sponding positions of the 30-isomer 20a, that is, compound 20c, re-
sulted in re-appearance of the activity, though it was weaker than
that of PP2P (9). Cinnamyl analog 22a showed very weak activity.
Among the compounds listed in Table 1, only PP2P (9), 20c and 22a
Overall, the structure–activity relationships for a-glucosidase-
inhibitory activity mentioned above are quite complex, though
some trends are apparent. Among the compounds prepared, 24c,
26a, 26b, and 27c showed rather potent
activities, which are comparable to that of PP2P (9), with IC50 val-
ues of lower than 20 M. Especially, 26a and 26b showed only very
a-glucosidase-inhibitory
were found to elicit
a-glucosidase-inhibitory activity under the
l
Table 1
a
-Glucosidase-inhibitory and LXR-antagonistic activities of phenethylphenyl phthalimides (9, 19–22)
4''
3''
R2
R1
O
2' 3'
N
4'
O
Compound
Position
Single (s) or double bond (d)
R1, R2
a
-Glucosidase IC50
(lM)
LXR
a
LXRb
% inhibition at 10
% inhibition at 10
l
M
IC50
(lM)
lM
IC50 (lM)
9
19b
19c
20a
20b
20c
21a
21b
21c
22a
22b
22c
20
20
20
30
30
30
40
40
40
40
40
40
s
s
s
s
s
s
s
s
s
d
d
d
H
16.2
>100
>100
>100
>100
38
>100
>100
>100
90
40
56
50
34
33
68
23
65
9.8
7.4
9.8
>30
>30
6.1
>30
4.8
1.7
>30
3.8
1.6
24
13
6
40
41
61
7
62
>30
>30
>30
>30
>30
7.6
OMe
OH
H
OMe
OH
H
OMe
OH
H
OMe
OH
>30
4.3
Toxica
30
Toxica
10
>3b
>30
>30
>1b
>100
>100
66
10
Toxica
Toxica
a
The cytotoxicity was observed when the assay was performed at 10
IC50 values could not be calculated because the cytotoxicity of this compound was observed at this concentration.
l
M.
b

K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
5005
Table 2
a
-Glucosidase-inhibitory and LXR-antagonistic activities of 300- or 400-substituted analogues of phenethylphenyl phthalimides (21b, 21c, 23)
O
4''
R2
N
3''
R1
O
Compound
R1
R2
a-Glucosidase IC50
(l
M)
LXR
a
LXRb
% inhibition at 10
lM
IC50
(l
M)
% inhibition at 10
lM
IC50 (lM)
21c
23e
23g
21b
23d
23f
OH
OH
H
OMe
OMe
H
OH
H
OH
OMe
H
>100
>100
>100
>100
>100
>100
Toxica
54
72
65
72
1.7
7.5
2.9
4.8
2.4
0.2
Toxica
23
45
62
75
>3b
>30
>30
4.3
3.5
>30
OMe
95
43
a
The cytotoxicity was observed when the assay was performed at 10
IC50 values could not be calculated because the cytotoxicity of this compound was observed at this concentration.
l
M.
b
Table 3
a
-Glucosidase-inhibitory and LXR-antagonistic activities of 4,5,6,7-tetrachlorophenethylphenyl phthalimides (24–30)
4''
R2
Cl
O
2' 3'
C
l
l
R1
N
4'
3''
C
O
Cl
Compound
Position
Single (s)
R1
R2
a-Glucosidase
LXR
a
LXRb
or double bond (d)
IC50 M)
(l
% inhibition at 10
lM
IC50
(lM)
% inhibition at 10
lM
IC50 (lM)
24a
24b
24c
25a
25b
25c
26a
26b
26c
27a
27b
27c
28a
28b
28c
29d
29f
20
20
20
30
30
30
40
40
40
30
30
30
40
40
40
30
30
30
30
40
40
40
40
s
s
s
s
s
s
s
s
s
d
d
d
d
d
d
s
s
s
s
s
s
s
s
H
H
>100
>100
20
>100
83
23
17
18
>100
31
27
13
21
40
>100
69
49
50
>100
65
Toxica
64
32
6
23
66
24
26
62
28
31
65
7
19
56
60
62
70
72
55
53
60
36
>3b
6.1
15
Toxica
57
26
0
11
21
18
6
22
9
8
24
12
8
26
17
18
9
>3b
7.7
17
>30
>30
21
OMe
OH
H
OMe
OH
H
OMe
OH
H
OMe
OH
H
OMe
OH
OMe
H
OH
H
OMe
H
OH
H
OMe
OH
H
OMe
OH
H
OMe
OH
H
OMe
OH
H
OMe
OH
H
OMe
H
OH
H
OMe
H
>30
>30
6.3
>30
>30
5.1
>30
>30
6.6
>30
>20
2.7
4.2
5.5
2.6
3.7
6.9
8.8
3.9
>30
>30
>30
>30
>30
>30
>30
>30
14
>30
>30
>30
>30
>30
>30
>30
>30
>30
29e
29g
30d
30f
30e
30g
21
30
12
18
36
>100
21
23
OH
a
Cytotoxicity was observed when the assay was performed at 10
IC50 values could not be calculated because the compound was cytotoxic at this concentration.
l
M.
b
weak LXR-antagonistic activities (Table 3, vide infra). To examine
were less potent than PP2P (9), with almost no LXRa/b-selectivity
the mode of
aver–Burk plot analysis was performed for 26b, riccardin C (5)
and the typical -glucosidase inhibitor deoxynojirimycin (Fig. 3).
Compound 26b was found to be a non-competitive inhibitor. We
confirmed that deoxynojirimycin42 and riccardin C (5)24 showed
competitive inhibition and non-competitive inhibition, respec-
tively, as reported.
a
-glucosidase inhibition elicited by 26b, Linewe-
(Table 1). Introduction of two methoxy or hydroxyl groups into
PP2P (9) had no apparent effect on the activity. However, introduc-
tion of two hydroxyl groups into 20a and 21a, that is, compounds
20c and 21c, respectively, resulted in a dramatic increase of the
a
activity for both LXRa and LXRb (Table 1). Introduction of two
methoxy groups into 21a, that is, compound 21b, also resulted in
increased activity, though the effects were less potent than those
of hydroxyl groups. On the contrary, introduction of two methoxy
groups into 20a, that is, 20b, had no apparent effect on the activity.
Among these dimethoxy and dihydroxyl derivatives, 40-substituted
analogs were more potent towards LXRs than 30-substituted ana-
logs (21b > 20b, and 21c > 20c). In the case of cinnamyl analogs
22a–c, substituent effects are much more apparent, especially for
2.3. Structure–activity relationship for LXR-antagonistic
activity
As previously reported, PP2P (9) showed slightly LXR
a-selective
antagonistic activity.24,34,35 Its regio-isomers (20a, 21a and 22a),

5006
K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
c
b
a
500
400
300
200
100
0
400
300
200
100
0
150
100
50
+26b
+Riccardin C
+deoxynojirimycin
(50 µM)
(30 µM)
(200 µM)
control
control
control
0
-0.2
0
0.2 0.4
0.6 0.8
1
1.2
-0.2
0
0.2 0.4 0.6 0.8
1/[S] (1/mM)
1
1.2
-0.4 -0.2
0
0.2 0.4 0.6 0.8
1/[S] (1/mM)
1
1.2
-100
-100
1/[S] (1/mM)
-50
Figure 3. Lineweaver–Burk plot analysis of the inhibition of a-glucosidase by (a) riccardin C (5), (b) deoxynojirimycin and (c) 26b.
LXR
tive. Although cytotoxicity appeared in dihydroxyl derivatives 21c
and 22c, these compounds were found to be potent LXR -selective
antagonists with IC50 values of 1.6–1.7 M, with almost no LXRb-
antagonistic or -glucosidase-inhibitory activity (Table 1).
Based on the results shown in Table 1, the 40-phenethyl series of
compounds, which lack -glucosidase-inhibitory activity, were se-
lected for further study (Table 2). Because deoxyriccardin C (7,
a
-antagonistic activity, with hydroxyl groups being more effec-
[26c > 26a > 26b, and 28c > 28a > 28b (Table 3)]. Concerning the
substituent effect elicited by hydroxyl groups, the 400-position
seems to be more effective than the 300-position (29g and 30g are
more potent than 29e and 30e, respectively), which is the reverse
a
l
a
of the order found for LXR
itory activities (vide supra). Finally, 28c and 29g are LXR
antagonists with IC50 values of 2.7 and 3.7 M for LXR , IC50 values
of >30 M for LXRb, and no apparent -glucosidase inhibitory
activity. Compounds 29d, 29e, 29f, 30d, 30e and 30f are also
LXR -selective antagonists (IC50 values of 2.6–8.8 M), but they
also possess -glucosidase-inhibitory activity (Table 3).
a
-antagonistic and
a-glucosidase-inhib-
a
-selective
a
l
a
l
a
Fig. 1) showed 11-fold selective antagonistic activity for LXRa over
LXRb,24 mono-deoxy or mono-demethoxy analogs of 21b/21c were
investigated. The mono-hydroxyl derivatives, 23e and 23g, were
a
l
a
less potent LXR
derivative 21c, but they retained selectivity for LXR
On the other hand, removal of one methoxy group from 21b re-
sulted in an increase of LXR -antagonistic activity, with removal
a
antagonists than the corresponding dihydroxyl
a
over LXRb.
3. Conclusion
a
Structural development studies of PP2P (9), which had been de-
rived from thalidomide and found to possess LXR /b-antagonistic
and -glucosidase-inhibitory activities, were performed. We were
able to separate the LXR-antagonistic and -glucosidase-inhibitory
activities, at least under the experimental conditions used here. A
potent LXR -selective antagonist 23f with very low LXRb-antago-
nistic activity (IC50 values of 0.2 M for LXR and >30 M for LXRb)
and no -glucosidase-inhibitory activity was obtained. The reason
why23fselectivelyantagonizesLXR isnotclear, butitis interesting
that both riccardin F (6) and compound 23f, each possessing a 4-
methoxyphenethyl group, showed LXR -selective antagonistic
activity. There is no difference in amino acid sequences between
LXR
and LXRb in the region of the ligand binding pocket;43,44 all
of the 300-methoxy group, that is, compound 23f, being more effec-
tive. Concerning LXRb-antagonistic activity, the effect of removal of
one methoxy group from 21b seems to be position-dependent: re-
moval of the 400-methoxy group (i.e., compound 23d) resulted in
slight increase in LXRb-antagonistic activity, while that of the 300-
methoxy group (i.e., compound 23f) resulted in a decrease of
LXRb-antagonistic activity. Finally, the 400-methoxy analog 23f pos-
a
a
a
a
l
a
l
a
sessed very potent LXR
of 0.2 M for LXR ) with more than 150-fold selectivity over LXRb.
Thus, substitution of the 300-methoxy group of the LXR
/b dual
-selective
a-selective antagonistic activity (IC50 value
a
l
a
a
a
antagonist 21b with hydrogen led to a potent LXR
a
antagonist, 23f. Its regio-isomer, the 300-methoxy analog 23d, was
a
found to be a rather potent non-selective LXRs dual antagonist
with IC50 values of 2.4–3.5 lM.
Concerning tetrachlorophthalimide derivatives (24–30, Table
3), the effects of tetrachlorination are not so clear as in the case
amino acid residues that are in close contact with known ligands
(i.e., within 5 Å distance from the molecular surface of the ligands)
are reported to be identical. However, there is one amino acid differ-
ence within helix-3 of the LBD (i.e., LXRb Ile277/LXR
possible interpretation of the LXR -selectivity of 23f might be an
interaction of the 4-methoxyphenethyl moiety of 23f with helix-3.
X-ray structural investigation of LXRs complexed with 23f seems
to be needed for a further consideration of the basis of the selectivity.
a
Val263).20 One
of
a
-glucosidase-inhibitory activity. Broadly speaking, tetrachlor-
-selective antagonis-
a
ophthalimide derivatives show slightly LXR
a
tic activity (their LXRb-antagonistic activities were not so potent,
or were absent, except for 24b). The substituent effects of dime-
thoxy and dihydroxyl groups (compounds 24–28, Table 3) are
complex, as was found in the case of
a-glucosidase-inhibitory
4. Experimental
4.1. Biology
activity. Concerning the LXR
a
-antagonistic activity of the 30-
(25a–c and 27a–c) and 40-phenethyl/cinnamyl (26a–c and 28a–c)
derivatives, the potency decreased in the order of: dihydr-
oxyl > dimethoxy > non-substituted derivatives [25c > 25b > 25a,
26c > 26b ꢁ 26a, 27c > 27b ꢁ 27a, and 28c > 28b > 28a (Table 3)].
Concerning the substituent effect elicited by hydroxyl groups, that
introduced at the 300-position seems to be more effective than that
at the 400-position (29e and 30e are more potent than 29g and 30g,
4.1.1.
a-Glucosidase inhibition assay
a-Glucosidase (Saccharomyces sp., Wako) 0.2 mU/mL in 10 mM
phosphate buffer (pH 7.0) was treated with DMSO solution of var-
ious compounds (final DMSO concentration 1% v/v) in a 96-well
plate (final volume 90
10 L pNPG solution (final concentration 0.2 mM) was added.
The mixture was incubated at 37 °C for 10 min, then basified by
adding 100 L of 0.5 M Na2CO3 solution. The amount of released
lL). After 10 min incubation at 37 °C,
respectively), as was the case for a-glucosidase-inhibitory activity
l
(vide supra). In the case of LXRb-antagonistic activity, the orders of
potency of 30-phenethyl/cinnamyl derivatives (25a–c and 27a–c)
l
are the same as those for LXRa-antagonistic activity. However,
p-nitrophenol was measured based on the absorbance at 405 nm.
The experiment was performed in triplicate and repeated at least
twice, and the normalized average values are presented. The IC50
values were reproducible.
those of the 40-phenethyl/cinnamyl derivatives (26a–c and 28a–
c) are different, that is, the order of potency decreased in the order
of:
dihydroxyl > non-substituted > dimethoxy
derivatives

K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
5007
4.1.2. Reporter gene assay
1H NMR (500 MHz, CDCl3) 11aZ d: 8.08–8.06 (m, 1H), 7.38–7.36
(m, 2H), 7.26–7.24 (m, 1H), 7.15–7.14 (m, 3H), 7.05–7.02 (m, 2H),
6.88 (d, 1H, J = 12.2 Hz), 6.75 (d, 1H, J = 12.2 Hz). Compound 11aE
d: 7.95 (dd, 1H, J = 7.9, 1.2 Hz), 7.75 (d, 1H, J = 7.9 Hz), 7.59 (d,
1H, J = 6.1 Hz), 7.57 (s, 1H), 7.53 (d, 2H, J = 7.3 Hz), 7.40–7.35 (m,
3H), 7.30 (t, 1H, J = 7.3 Hz), 7.07 (d, 1H, J = 16.5 Hz). FAB-MS m/z:
225 [M]+, 226 [M+H]+.
Human embryonic kidney (HEK) 293 cells were cultured in Dul-
becco’s modified Eagle’s medium containing 5% fetal bovine serum
at 37 °C in a humidified atmosphere of 5% CO2 in air. Transfections
were performed by the calcium phosphate coprecipitation method.
Test compounds with or without 0.1 lM 1 were added 8 h after the
transfection, and luciferase and b-galactosidase activities were as-
sayed using a luminometer and microplate reader. The experiment
was performed in triplicate and repeated at least twice, and the
normalized average values are presented. The IC50 values were
reproducible.
4.2.5. 1,2-Dimethoxy-4-[2-(2-nitrophenyl)ethenyl]benzene
(11b)
This compound was prepared from 2-nitrobenzaldehyde and
10b by means of a procedure similar to that used for 11a. Com-
pound 11b was obtained in 95% yield as a brown oil.
1H NMR (500 MHz, CDCl3) 11bZ d: 8.08 (dd, 1H, J = 7.9, 1.2 Hz),
7.45–7.42 (td, 1H, J = 7.3, 1.2 Hz), 7.41–7.37 (td, 1H, J = 7.9, 1.8 Hz),
7.35 (d, 1H, J = 7.3 Hz), 6.81 (d, 1H, J = 12.2 Hz), 6.70–6.66 (m, 3H),
6.50 (d, 1H, J = 1.8 Hz), 3.83 (s, 3H), 3.55 (s, 3H). FAB-MS m/z: 285
[M]+, 286 [M+H]+.
4.2. General
Melting points were determined by using a Yanagimoto hot-
stage melting point apparatus and are uncorrected. 1H NMR spec-
tra were recorded on a JEOL JNMGX500 (500 MHz) spectrometer.
Chemical shifts are expressed in parts per million relative to tetra-
methylsilane. Mass spectra were recorded on a JEOL JMS-DX303
spectrometer.
4.2.6. 1-Nitro-3-(2-phenylethenyl)benzene (12a)
To a solution of 3-nitrobenzaldehyde (680 mg, 4.5 mmol) in
dehydrated dichloromethane (20 mL) were added benzyltriphenyl-
phosphonium chloride (1.75 g, 4.5 mmol), potassium carbonate
(691 mg, 5.0 mmol) and 18-crown-6 (211 mg, 0.8 mmol). The mix-
ture was refluxed for 5 h, then filtered and concentrated. The resi-
due was purified by silica gel column chromatography to give the
target compound (994 mg, 98%) as a pale yellow solid. Addition-
ally, the EZ mixture was purified by silica gel column chromatogra-
phy to afford 12aE (32%) as a pale yellow solid.
4.2.1. 3,4-Dimethoxybenzyltriphenylphosphonium bromide (10b)
To a solution of 3,4-dimethoxytoluene (2.90 mL, 20 mmol) in
carbon tetrachloride (40 mL) were added N-bromosuccinimide
(3.92 g, 22 mmol) and 2,20-azobis(isobutyronitrile) (476 mg,
2.9 mmol). The mixture was refluxed for 1 h. The mixture was fil-
tered and concentrated. The residue was purified by silica gel col-
umn chromatography to give an intermediate. To a solution of the
intermediate (5.89 g, 25.4 mmol) in acetonitrile (40 mL) was added
triphenylphosphine (9.99 g, 38.1 mmol). The mixture was refluxed
for 45 min. The reaction mixture was concentrated and mixed with
toluene. The whole was filtered to give the target compound
(8.07 g, 82%) as a pale yellow solid.
1H NMR (500 MHz, CDCl3) 12aZ d: 8.08 (t, 1H, J = 1.8 Hz), 8.03
(dd, 1H, J = 8.5, 1.8 Hz), 7.52 (d, 1H, J = 7.9 Hz), 7.35 (t, 1H, J = 7.9
Hz), 7.25-7.23 (m, 3H), 7.20-7.18 (m, 2H), 6.78 (d, 1H, J = 12.2
Hz), 6.61 (d, 1H, J = 12.2 Hz). Compound 12aE d: 8.35 (t, 1H,
J = 1.8 Hz), 8.09–8.07 (m, 1H), 7.78 (d, 1H, J = 7.9 Hz), 7.53 (m,
1H), 7.52–7.49 (m, 2H), 7.38 (t, 2H, J = 7.3 Hz), 7.30 (tt, 1H,
J = 7.3, 1.8 Hz), 7.22 (d, 1H, J = 15.9 Hz), 7.12 (d, 1H, J = 15.9 Hz).
FAB-MS m/z: 225 [M]+, 226 [M+H]+.
1H NMR (500 MHz, CDCl3) d: 7.79–7.73 (m, 9H), 7.66–7.62 (m,
6H), 6.85 (s, 1H), 6.62 (s, 2H), 5.36 (d, 2H, J = 14.0 Hz), 3.81 (s,
3H), 3.55 (s, 3H). FAB-MS m/z: 413 [MꢂBr]+.
4.2.2. 3-Methoxybenzyltriphenylphosphonium bromide (10d)
To a solution of 3-methoxybenzyl bromide (700
l
L, 5.0 mmol) in
4.2.7. 1,2-Dimethoxy-4-[2-(3-nitrophenyl)ethenyl]benzene (12b)
This compound was prepared from 3-nitrobenzaldehyde and
10b by means of a procedure similar to that used for 12a. Com-
pound 12b was obtained in 95% yield as a brown oil. Compound
12bE was obtained in 40% yield as a yellow solid.
acetonitrile (20 mL) was added triphenylphosphine (3.72 g,
14.2 mmol). The mixture was refluxed for 45 min. The reaction mix-
ture was concentrated and mixed with toluene. The whole was fil-
tered to give the target compound (2.19 g, 94%) as a white solid.
1H NMR (500 MHz, CDCl3) d: 7.79–7.73 (m, 9H), 7.66–7.62 (m,
6H), 7.03 (t, 1H, J = 7.9 Hz), 6.82–6.80 (m, 1H), 6.77 (dt, 1H,
J = 8.5, 2.4 Hz), 6.63 (d, 1H, J = 7.3 Hz), 5.38 (d, 2H, J = 14.6 Hz),
3.55 (s, 3H). FAB-MS m/z: 383 [MꢂBr]+.
1H NMR (500 MHz, CDCl3) 12bZ d: 8.15 (t, 1H, J = 1.8 Hz), 8.04
(dd, 1H, J = 7.3, 2.4 Hz), 7.59 (d, 1H, J = 7.3 Hz), 7.39 (t, 1H, J = 7.9
Hz), 6.80-6.75 (m, 2H), 6.71 (s, 1H), 6.70 (d, 1H, J = 12.2 Hz), 6.53
(d, 1H, J = 12.2 Hz), 3.87 (s, 3H), 3.65 (s, 3H). Compound 12bE d:
8.35 (t, 1H, J = 1.8 Hz), 8.08–8.06 (m, 1H), 7.78 (d, 1H, J = 7.9 Hz),
7.51 (t, 1H, J = 7.9 Hz), 7.19 (d, 1H, J = 15.9 Hz), 7.10 (dd, 1H,
J = 7.3, 1.8 Hz), 7.09 (s, 1H), 7.01 (d, 1H, J = 15.9 Hz), 6.89 (d, 1H,
J = 9.2 Hz), 3.96 (s, 3H), 3.92 (s, 3H). FAB-MS m/z: 285 [M]+, 286
[M+H]+.
4.2.3. 4-Methoxybenzyltriphenylphosphonium bromide (10f)
This compound was prepared from 4-methoxybenzyl bromide
by means of a procedure similar to that used for 10d. Compound
10f was obtained in 91% yield as a white solid.
1H NMR (500 MHz, CDCl3) d: 7.79–7.72 (m, 9H), 7.66–7.62 (m,
6H), 7.03 (dd, 2H, J = 8.5, 2.4 Hz), 6.66 (d, 2H, J = 8.5 Hz), 5.33 (d,
2H, J = 14.0 Hz), 3.73 (s, 3H). FAB-MS m/z: 383 [MꢂBr]+.
4.2.8. 1-Methoxy-3-[2-(3-nitrophenyl)ethenyl]benzene (12d)
This compound was prepared from 3-nitrobenzaldehyde and
10d by means of a procedure similar to that used for 11a. Com-
pound 12d was obtained in 93% yield as a yellow oil.
4.2.4. 1-Nitro-2-(2-phenylethenyl)benzene (11a)
To a solution of 2-nitrobenzaldehyde (302 mg, 2.0 mmol) in
dehydrated dichloromethane (12 mL) were added benzyltriphenyl-
phosphonium chloride (778 mg, 2.0 mmol), potassium carbonate
(304 mg, 2.2 mmol) and 18-crown-6 (95 mg, 0.36 mmol). The mix-
ture was refluxed for 9 h, then filtered and concentrated. The resi-
due was purified by silica gel column chromatography to give the
target compound (410 mg, 91%) as a yellow oil. Additionally, the EZ
mixture was purified by silica gel column chromatography to pro-
vide a sample for instrumental analysis.
1H NMR (500 MHz, CDCl3) 12dZ d: 8.09 (t, 1H, J = 1.8 Hz), 8.02
(dd, 1H, J = 7.9, 1.2 Hz), 7.54 (d, 1H, J = 7.9 Hz), 7.35 (t, 1H,
J = 7.9 Hz), 7.16 (t, 1H, J = 7.9 Hz), 6.79–6.73 (m, 3H), 6.74 (d, 1H,
J = 12.2 Hz), 6.59 (d, 1H, J = 12.2 Hz), 3.68 (s, 3H). Compound
12dE d: 8.37 (t, 1H, J = 1.8 Hz), 8.10 (dd, 1H, J = 8.5, 1.8 Hz), 7.80
(d, 1H, J = 7.9 Hz), 7.53 (t, 1H, J = 7.9 Hz), 7.31 (t, 1H, J = 7.9 Hz),
7.21 (d, 1H, J = 16.5 Hz), 7.14 (d, 1H, J = 7.3 Hz), 7.13 (d, 1H,
J = 16.5 Hz), 7.07 (t, 1H, J = 1.8 Hz), 6.89 (dd, 1H, J = 8.5, 2.4 Hz),
3.87 (s, 3H). FAB-MS m/z: 255 [M]+, 256 [M+H]+.

5008
K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
4.2.9. 1-Methoxy-4-[2-(3-nitrophenyl)ethenyl]benzene (12f)
This compound was prepared from 3-nitrobenzaldehyde and
10f by means of a procedure similar to that used for 11a. Com-
pound 12f was obtained in 92% yield as a yellow solid.
mixture was filtered through a pad of Celite, and the filtrate was
evaporated under reduced pressure. The residue was purified by
silica gel column chromatography to give the target compound
(366 mg, 100%) as a colorless oil.
1H NMR (500 MHz, CDCl3) 12fZ d: 8.12 (t, 1H, J = 1.8 Hz), 8.04–
8.02 (m, 1H), 7.56 (d, 1H, J = 7.9 Hz), 7.37 (t, 1H, J = 7.9 Hz), 7.13 (d,
2H, J = 8.5 Hz), 6.77 (d, 2H, J = 8.5 Hz), 6.70 (d, 1H, J = 12.2 Hz), 6.51
(d, 1H, J = 12.2 Hz), 3.79 (s, 3H). Compound 12fE d: 8.34 (t, 1H,
J = 1.8 Hz), 8.08–8.06 (m, 1H), 7.76 (d, 1H, J = 7.3 Hz), 7.50 (t, 1H,
J = 7.9 Hz), 7.49 (d, 2H, J = 8.5 Hz), 7.19 (d, 1H, J = 16.2 Hz), 7.00
(d, 1H, J = 16.2 Hz), 6.93 (d, 2H, J = 8.5 Hz), 3.85 (s, 3H). FAB-MS
m/z: 255 [M]+, 256 [M+H]+.
1H NMR (500 MHz, CDCl3) d: 7.31–7.28 (m, 2H), 7.23–7.20 (m,
3H), 7.07–7.04 (m, 2H), 6.75 (td, 1H, J = 7.3, 1.2 Hz), 6.68 (dd, 1H,
J = 7.3, 1.2 Hz), 3.50 (br s, 2H), 2.96–2.92 (m, 2H), 2.81–2.78 (m,
2H). FAB-MS m/z: 197 [M]+, 198 [M+H]+.
4.2.15. 2-[2-(3,4-Dimethoxyphenyl)ethyl]aniline (14b)
This compound was prepared from 11b by means of a proce-
dure similar to that used for 14a. Compound 14b was obtained
in 92% yield as a colorless oil.
1H NMR (500 MHz, CDCl3) d: 7.06–7.02 (m, 2H), 6.80 (d, 1H,
J = 7.9 Hz), 6.76–6.73 (m, 2H), 6.67 (d, 1H, J = 7.9 Hz), 6.63 (d, 1H,
J = 1.8 Hz), 3.86 (s, 3H), 3.81 (s, 3H), 3.47 (br s, 2H), 2.90–2.86 (m,
2H), 2.79–2.76 (m, 2H). FAB-MS m/z: 257 [M]+, 258 [M+H]+.
4.2.10. 1-Nitro-4-(2-phenylethenyl)benzene (13a)
This compound was prepared from 4-nitrobenzaldehyde and
10a by means of a procedure similar to that used for 12a. Com-
pound 13a was obtained in 86% yield as yellow solid. Compound
13aE was obtained in 23% yield as a yellow solid.
1H NMR (500 MHz, CDCl3) 13aZ d: 8.07 (d, 2H, J = 8.9 Hz), 7.37
(d, 2H, J = 8.9 Hz), 7.27–7.25 (m, 3H), 7.21–7.19 (m, 2H), 6.82 (d,
1H, J = 12.2 Hz), 6.62 (d, 1H, J = 12.2 Hz). Compound 13aE d: 8.23
(dt, 2H, J = 9.2, 2.4 Hz), 7.64 (dt, 2H, J = 9.2, 2.4 Hz), 7.56 (d, 2H,
J = 7.3 Hz), 7.40 (d, 2H, J = 7.3 Hz), 7.34 (tt, 1H, J = 7.3, 2.4 Hz),
7.28 (d, 1H, J = 16.5 Hz), 7.15 (d, 1H, J = 16.5 Hz). FAB-MS m/z:
225 [M]+, 226 [M+H]+.
4.2.16. 3-(2-Phenylethyl)aniline (15a)
This compound was prepared from 12a by means of a procedure
similar to that used for 14a. Compound 15a was obtained in 96%
yield as a white solid.
1H NMR (500 MHz, CDCl3) d: 7.28 (s, 1H), 7.26 (d, 1H, J = 7.9 Hz),
7.20–7.18 (m, 3H), 7.09–7.06 (m, 1H), 6.61 (d, 1H, J = 7.9 Hz), 6.54–
6.53 (m, 2H), 3.57 (br s, 2H), 2.92–2.88 (m, 2H), 2.85–2.81 (m, 2H).
FAB-MS m/z: 197 [M]+, 198 [M+H]+.
4.2.11. 1,2-Dimethoxy-4-[2-(4-nitrophenyl)ethenyl]benzene (13b)
This compound was prepared from 4-nitrobenzaldehyde and
10b by means of a procedure similar to that used for 12a. Com-
pound 13b was obtained in 98% yield as a yellow solid. Compound
13bE was obtained in 46% yield as a yellow solid.
4.2.17. 3-[2-(3,4-Dimethoxyphenyl)ethyl]aniline (15b)
This compound was prepared from 12b by means of a proce-
dure similar to that used for 14a. Compound 15b was obtained
in 74% yield as a white solid.
1H NMR (500 MHz, CDCl3) 13bZ d: 8.09 (d, 2H, J = 8.9 Hz), 7.43
(d, 2H, J = 8.9 Hz), 6.80–6.71 (m, 4H), 6.53 (d, 1H, J = 12.2 Hz),
3.88 (s, 3H), 3.67 (s, 3H). Compound 13bE d: 8.21 (d, 2H,
J = 8.5 Hz), 7.61 (d, 2H, J = 8.5 Hz), 7.22 (d, 2H, J = 16.5 Hz), 7.11
(dd, 1H, J = 7.9, 1.8 Hz), 7.10 (d, 1H, J = 1.8 Hz), 7.01 (d, 1H,
J = 16.5 Hz), 6.90 (d, 1H, J = 7.9 Hz), 3.96 (s, 3H), 3.92 (s, 3H). FAB-
MS m/z: 285 [M]+, 286 [M+H]+.
1H NMR (500 MHz, CDCl3) d: 7.07 (td, 1H, J = 7.3, 1.2 Hz), 6.79
(d, 1H, J = 7.9 Hz), 6.73 (dd, 1H, J = 7.9, 1.8 Hz), 6.67 (d, 1H,
J = 1.8 Hz), 6.60 (d, 1H, J = 7.9 Hz), 6.54–6.52 (m, 1H), 6.52 (d, 1H,
J = 1.2 Hz), 3.86 (s, 3H), 3.84 (s, 3H), 2.87–2.83 (m, 2H), 2.82–2.79
(m, 2H). FAB-MS m/z: 257 [M]+, 258 [M+H]+.
4.2.18. 3-[2-(3-Methoxyphenyl)ethyl]aniline (15d)
This compound was prepared from 12d by means of a proce-
dure similar to that used for 14a. Compound 15d was obtained
in 74% yield as a colorless oil.
4.2.12. 1-Methoxy-3-[2-(4-nitrophenyl)ethenyl]benzene (13d)
This compound was prepared from 4-nitrobenzaldehyde and
10d by means of a procedure similar to that used for 11a. Com-
pound 13d was obtained in 100% yield as a yellow oil.
1H NMR (500 MHz, CDCl3) 13dZ d: 8.08 (d, 2H, J = 8.5 Hz), 7.39
(d, 2H, J = 8.5 Hz), 7.18 (t, 1H, J = 7.9 Hz), 6.81–6.74 (m, 4H), 6.61
(d, 1H, J = 12.2 Hz), 3.70 (s, 3H). Compound 13dE d: 8.22 (d, 2H,
J = 8.5 Hz), 7.64 (d, 2H, J = 8.5 Hz), 7.32 (t, 1H, J = 7.9 Hz), 7.24 (d,
1H, J = 16.5 Hz), 7.14 (d, 1H, J = 16.5 Hz), 7.08 (t, 1H, J = 1.8 Hz),
6.89 (dd, 1H, J = 7.9, 2.4 Hz), 3.86 (s, 3H). FAB-MS m/z: 255 [M]+,
256 [M+H]+.
1H NMR (500 MHz, CDCl3) d: 7.22–7.18 (m, 1H), 7.09–7.06 (m,
1H), 6.79 (d, 1H, J = 7.9 Hz), 6.75–6.73 (m, 2H), 6.62–6.60 (d, 1H,
J = 7.3 Hz), 6.53 (s, 1H), 6.53–6.52 (m, 1H), 3.78 (s, 3H), 3.59 (br s,
2H), 2.89–2.85 (m, 2H), 2.83–2.80 (m, 2H). FAB-MS m/z: 227
[M]+, 228 [M+H]+.
4.2.19. 3-[2-(4-Methoxyphenyl)ethyl]aniline (15f)
This compound was prepared from 12f by means of a procedure
similar to that used for 14a. Compound 15f was obtained in 91%
yield as a white solid.
4.2.13. 1-Methoxy-4-[2-(4-nitrophenyl)ethenyl]benzene (13f)
This compound was prepared from 4-nitrobenzaldehyde and
10f by means of a procedure similar to that used for 11a. Com-
pound 13f was obtained in 94% yield as a yellow solid.
1H NMR (500 MHz, CDCl3) d: 7.10 (d, 2H, J = 8.5 Hz), 7.09–7.05
(m, 1H), 6.83 (d, 2H, J = 8.5 Hz), 6.60 (d, 1H, J = 7.9 Hz), 6.54–6.53
(m, 2H), 3.79 (s, 3H), 3.60 (br s, 2H), 2.86–2.82 (m, 2H), 2.81–
2.77 (m, 2H). FAB-MS m/z: 227 [M]+, 228 [M+H]+.
1H NMR (500 MHz, CDCl3) 13fZ d: 8.08 (d, 2H, J = 9.2 Hz), 7.40
(d, 2H, J = 8.5 Hz), 7.14 (d, 2H, J = 8.5 Hz), 6.78 (d, 2H, J = 9.8 Hz),
6.73 (d, 1H, J = 12.2 Hz), 6.51 (d, 1H, J = 12.2 Hz), 3.80 (s, 3H).
Compound 13fE d: 8.20 (d, 2H, J = 8.5 Hz), 7.60 (d, 2H,
J = 8.5 Hz), 7.50 (d, 2H, J = 8.5 Hz), 7.23 (d, 1H, J = 16.5 Hz), 7.01
(d, 1H, J = 16.5 Hz), 6.93 (d, 2H, J = 8.5 Hz), 3.85 (s, 3H). FAB-MS
m/z: 255 [M]+, 256 [M+H]+.
4.2.20. 4-(2-Phenylethyl)aniline (16a)
This compound was prepared from 13a by means of a procedure
similar to that used for 14a. Compound 16a was obtained in 95%
yield as a white solid.
1H NMR (500 MHz, CDCl3) d: 7.29–7.26 (m, 2H), 7.20–7.17 (m,
3H), 6.97 (d, 2H, J = 8.0 Hz), 6.63 (d, 2H, J = 8.0 Hz), 3.55 (br s,
2H), 2.88–2.86 (m, 2H), 2.83–2.79 (m, 2H). FAB-MS m/z: 197
[M]+, 198 [M+H]+.
4.2.14. 2-(2-Phenylethyl)aniline (14a)
Compound 11a (410 mg, 1.82 mmol) was dissolved in EtOAc
(20 mL) and hydrogenated with 10% Pd/C (catalytic amount). The

K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
5009
4.2.21. 4-[2-(3,4-Dimethoxyphenyl)ethyl]aniline (16b)
This compound was prepared from 13b by means of a proce-
dure similar to that used for 14a. Compound 16b was obtained
in 100% yield as a white solid.
1H NMR (500 MHz, CDCl3) d: 7.32 (d, 2H, J = 8.5 Hz), 7.04 (d, 1H,
J = 1.8 Hz), 7.00 (dd, 1H, J = 7.9, 1.8 Hz), 6.88 (m, 1H), 6.88 (m, 1H),
6.84 (d, 1H, J = 7.9 Hz), 6.68 (d, 2H, J = 8.5 Hz), 3.94 (s, 3H), 3.90 (s,
3H), 3.73 (br s, 2H). FAB-MS m/z: 255 [M]+, 256 [M+H]+.
1H NMR (500 MHz, CDCl3) d: 6.96 (d, 2H, J = 8.5 Hz), 6.78 (d, 1H,
J = 7.9 Hz), 6.71 (dd, 1H, J = 7.9, 1.8 Hz), 6.65 (d, 1H, J = 1.8 Hz), 6.62
(d, 2H, J = 8.5 Hz), 3.86 (s, 3H), 3.84 (s, 3H), 3.57 (br s, 2H), 2.82–
2.78 (m, 4H). FAB-MS m/z: 257 [M]+, 258 [M+H]+.
4.2.28. N-{2-[2-(3,4-Dimethoxyphenyl)ethyl]phenyl}phthalimide
(19b)
A mixture of phthalic anhydride (87 mg, 0.59 mmol) and 14b
(151 mg, 0.59 mmol) was heated at 200 °C for 1 h. After the reac-
tion was completed, the residue was purified by silica gel column
chromatography to give the target compound (201 mg, 88%) as a
white solid.
4.2.22. 4-[2-(3-Methoxyphenyl)ethyl]aniline (16d)
This compound was prepared from 13d by means of a proce-
dure similar to that used for 14a. Compound 16d was obtained
in 93% yield as a white solid.
Colorless needles from CH2Cl2/n-hexane. Mp 103.0–105.0 °C. 1H
NMR (500 MHz, CDCl3) d: 7.95 (dd, 2H, J = 5.5, 3.1 Hz), 7.80 (dd, 2H,
J = 5.5, 3.1 Hz), 7.41–7.37 (m, 1H), 7.37–7.33 (m, 1H), 7.31 (dd, 1H,
J = 7.3, 1.8 Hz), 7.20 (dd, 1H, J = 7.3, 1.8 Hz), 6.69 (d, 1H, J = 7.9 Hz),
6.59 (dd, 1H, J = 7.9, 1.8 Hz), 6.50 (d, 1H, J = 1.8 Hz), 3.80 (s, 3H),
3.68 (s, 3H), 2.79 (s, 4H). FAB-MS m/z: 387 [M]+, 388 [M+H]+. Anal.
Calcd for C24H21NO4: C, 74.40; H, 5.46; N, 3.62. Found: C, 74.34; H,
5.59; N, 3.55.
1H NMR (500 MHz, CDCl3)d: 7.19 (td, 1H, J = 7.6, 1.2 Hz), 6.98(d, 2H,
J = 8.5 Hz), 6.78 (d, 1H, J = 7.3 Hz), 6.73 (dd, 1H, J = 7.3, 1.8 Hz), 6.73 (s,
1H), 6.63 (d, 2H, J = 7.9 Hz), 3.78 (s, 3H), 3.55 (br s, 2H), 2.86–2.82 (m,
2H), 2.82–2.78 (m, 2H). FAB-MS m/z: 227 [M]+, 228 [M+H]+.
4.2.23. 4-[2-(4-Methoxyphenyl)ethyl]aniline (16f)
This compound was prepared from 16f by means of a procedure
similar to that used for 14a. Compound 16f was obtained in 91%
yield as a white solid.
4.2.29. N-{2-[2-(3,4-Dihydroxyphenyl)ethyl]phenyl}phthalimide
(19c)
1H NMR (500 MHz, CDCl3) d: 7.08 (d, 2H, J = 8.5 Hz), 6.96 (d, 2H,
J = 7.9 Hz), 6.82 (d, 2H, J = 8.5 Hz), 6.62 (d, 2H, J = 7.9 Hz), 3.79 (s,
3H), 3.55 (br s, 2H), 2.83–2.79 (m, 2H), 2.78–2.75 (m, 2H). FAB-
MS m/z: 227 [M]+, 228 [M+H]+.
To a solution of 19b (88 mg, 0.23 mmol) in dehydrated dichlo-
romethane (2 mL) was added dropwise boron tribromide (1.0 M
solution in dichloromethane) (2.3 mL, 2.30 mmol) at 0 °C under
an argon atmosphere. The mixture was stirred for 30 min, then
poured into water (20 mL) and extracted with CH2Cl2
(20 mL ꢃ 3). The organic layer was washed with H2O (20 mL)
and then dried over MgSO4. The solvent was removed under re-
duced pressure. The residue was purified by silica gel column
chromatography to give the target compound (75 mg, 90%) as a
white solid.
4.2.24. 3-[(1E)-2-Phenylethenyl]aniline (17a)
To a solution of 12aE (73 mg, 0.32 mmol) in EtOAc (10 mL) was
added tin (II) chloride dihydrate (361 mg, 1.60 mmol) and the mix-
ture was refluxed for 5 h. The reaction was then terminated by the
addition of satd NaHCO3 aq. The resulting mixture was filtered
through a pad of Celite and extracted with EtOAc (100 mL ꢃ 2).
The organic layer was washed with H2O (30 mL) and brine
(20 mL) and then dried over MgSO4. The solvent was removed un-
der reduced pressure. The residue was purified by silica gel column
chromatography to give the target compound (50 mg, 81%) as a
pale yellow solid.
Mp 63.0–65.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.95 (dd, 2H,
J = 5.5, 2.4 Hz), 7.81 (dd, 2H, J = 5.5, 2.4 Hz), 7.41–7.38 (m, 1H),
7.36–7.31 (m, 2H), 7.18 (dd, 1H, J = 7.9, 1.2 Hz), 6.66 (d, 1H,
J = 7.9 Hz), 6.52 (d, 1H, J = 1.8 Hz), 6.49 (dd, 1H, J = 7.9, 1.8 Hz),
5.16 (s, 1H), 5.00 (s, 1H), 2.76 (m, 4H). FAB-MS m/z: 359 [M]+,
360 [M+H]+. HRMS (FAB) calcd for C22H17NO4 359.1158; found:
359.1127 (M)+.
1H NMR (500 MHz, CDCl3) d: 7.50 (d, 2H, J = 7.3 Hz), 7.35 (t, 2H,
J = 7.3 Hz), 7.24 (d, 1H, J = 7.3 Hz), 7.15 (t, 1H, J = 7.9 Hz), 7.07 (d,
1H, J = 16.5 Hz), 7.02 (d, 1H, J = 16.5 Hz), 6.94 (d, 1H, J = 7.9 Hz),
6.85 (t, 1H, J = 1.8 Hz), 6.61 (dd, 1H, J = 7.9, 2.4 Hz), 3.68 (br s,
2H). FAB-MS m/z: 195 [M]+, 196 [M+H]+.
4.2.30. N-[3-(2-Phenylethyl)phenyl]phthalimide (20a)
This compound was prepared from 15a by means of a procedure
similar to that used for 19b. Compound 20a was obtained in 83%
yield as colorless needles after recrystallization from CH2Cl2/n-
hexane.
4.2.25. 3-[(1E)-2-(3,4-Dimethoxyphenyl)ethenyl]aniline (17b)
This compound was prepared from 12bE by means of a proce-
dure similar to that used for 17a. Compound 17b was obtained
in 76% yield as a white solid.
Mp 98.0–98.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 2.4 Hz), 7.80 (dd, 2H, J = 5.5, 2.4 Hz), 7.42 (t, 1H,
J = 7.9 Hz), 7.31–7.28 (m, 4H), 7.23–7.20 (m, 4H), 3.02–2.94 (m,
4H). FAB-MS m/z: 327 [M]+, 328 [M+H]+. Anal. Calcd for
C22H17NO2ꢀ1/4H2O: C, 79.62; H, 5.31; N, 4.22. Found: C, 79.50; H,
5.34; N, 4.19.
1H NMR (500 MHz, CDCl3) d: 7.14 (t, 1H, J = 7.9 Hz), 7.06 (d, 1H,
J = 1.8 Hz), 7.04 (dd, 1H, J = 7.9, 1.8 Hz), 7.01 (d, 1H, J = 16.8 Hz),
6.92 (d, 1H, J = 7.9 Hz), 6.89 (d, 1H, J = 16.8 Hz), 6.86 (d, 1H,
J = 8.5 Hz), 6.84 (m, 1H), 6.59 (dd, 1H, J = 7.9, 2.4 Hz), 3.95 (s, 3H),
3.90 (s, 3H), 3.67 (br s, 2H). FAB-MS m/z: 255 [M]+, 256 [M+H]+.
4.2.26. 4-[(1E)-2-Phenylethenyl]aniline (18a)
4.2.31. N-{3-[2-(3,4-Dimethoxyphenyl)ethyl]phenyl}phthalimide
(20b)
This compound was prepared from 15b by means of a proce-
dure similar to that used for 19b. Compound 20b was obtained
in 71% yield as colorless needles after recrystallization from
CH2Cl2/n-hexane.
This compound was prepared from 13aE by means of a proce-
dure similar to that used for 17a. Compound 18a was obtained in
83% yield as a white solid.
1H NMR (500 MHz, CDCl3) d: 7.47 (d, 2H, J = 7.3 Hz), 7.35–7.31
(m, 4H), 7.21 (t, 1H, J = 7.3 Hz), 7.03 (d, 1H, J = 16.2 Hz), 6.92 (d,
1H, J = 16.2 Hz), 6.68 (d, 2H, J = 8.5 Hz), 3.74 (br s, 2H). FAB-MS
m/z: 195 [M]+, 196 [M+H]+.
Mp 132.5–133.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 3.1 Hz), 7.80 (dd, 2H, J = 5.5, 3.1 Hz), 7.43–7.39 (m, 1H),
7.28–7.26 (m, 2H), 7.19 (d, 1H, J = 7.3 Hz), 6.80 (d, 1H, J = 7.9 Hz),
6.73 (dd, 1H, J = 7.9, 1.8 Hz), 6.65 (d, 1H, J = 1.8 Hz), 3.85 (s, 3H),
3.83 (s, 3H), 2.99–2.95 (m, 2H), 2.93–2.89 (m, 2H). FAB-MS m/z:
387 [M]+, 388 [M+H]+. Anal. Calcd for C24H21NO4: C, 74.40; H,
5.46; N, 3.62. Found: C, 74.33; H, 5.49; N, 3.54.
4.2.27. 4-[(1E)-2-(3,4-Dimethoxyphenyl)ethenyl]aniline (18b)
This compound was prepared from 13bE by means of a proce-
dure similar to that used for 17a. Compound 18b was obtained
in 88% yield as a yellow solid.

5010
K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
4.2.32. N-{3-[2-(3,4-Dihydroxyphenyl)ethyl]phenyl}phthalimide
(20c)
4.2.37. N-{4-[(1E)-2-(3,4-Dimethoxyphenyl)ethenyl]phenyl}ph-
thalimide (22b)
This compound was prepared from 20b by means of a proce-
dure similar to that used for 19c. Compound 20c was obtained in
70% yield as yellow needles after recrystallization from EtOAc/n-
hexane.
This compound was prepared from 18a by means of a procedure
similar to that used for 19b. Compound 22b was obtained in 64%
yield as yellow needles after recrystallization from CH2Cl2/n-
hexane.
Mp 171.5–172.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 2.4 Hz), 7.81 (dd, 2H, J = 5.5, 2.4 Hz), 7.42 (t, 1H,
J = 7.9 Hz), 7.23 (dd, 1H, J = 7.9, 2.4 Hz), 7.22 (dd, 1H, J = 7.9,
1.8 Hz), 7.13 (m, 1H), 6.78 (d, 1H, J = 7.9 Hz), 6.63 (dd, 1H, J = 7.9,
1.8 Hz), 6.56 (d, 1H, J = 1.8 Hz), 5.66 (s, 1H), 5.16 (s, 1H), 2.94–
2.90 (m, 2H), 2.86–2.83 (m, 2H). FAB-MS m/z: 359 [M]+, 360
[M+H]+. Anal. Calcd for C22H17NO4: C, 73.53; H, 4.77; N, 3.90.
Found: C, 73.24; H, 4.84; N, 3.83.
Mp 224.0–226.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.97 (dd, 2H,
J = 5.5, 3.1 Hz), 7.81 (dd, 2H, J = 5.5, 3.1 Hz), 7.63 (d, 2H,
J = 8.5 Hz), 7.45 (d, 2H, J = 8.5 Hz), 7.10 (d, 1H, J = 16.2 Hz), 7.09–
7.05 (m, 2H), 7.01 (d, 1H, J = 16.2 Hz), 6.88 (d, 1H, J = 7.9 Hz),
3.97 (s, 3H), 3.92 (s, 3H). FAB-MS m/z: 385 [M]+, 386 [M+H]+. Anal.
Calcd for C24H19NO4ꢀ1/3H2O: C, 73.64; H, 5.06; N, 3.58. Found: C,
73.83; H, 5.01; N, 3.30.
4.2.38. N-{4-[(1E)-2-(3,4-Dihydroxyphenyl)ethenyl]phenyl}ph-
thalimide (22c)
This compound was prepared from 22b by means of a proce-
dure similar to that used for 19c. Compound 22c was obtained in
44% yield as yellow needles after recrystallization from EtOAc/n-
hexane.
4.2.33. N-(4-(2-Phenylethyl)phenyl)phthalimide (21a)
This compound was prepared from 16a by means of a procedure
similar to that used for 19b. Compound 21a was obtained in 89%
yield as colorless needles after recrystallization from CH2Cl2/n-
hexane.
Mp 216.5–218.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 3.1 Hz), 7.79 (dd, 2H, J = 5.5, 3.1 Hz), 7.36–7.29 (m, 6H),
7.23–7.19 (m, 3H), 3.01–2.94 (m, 4H). FAB-MS m/z: 327 [M]+,
328 [M+H]+. Anal. Calcd for C22H17NO2: C, 80.71; H, 5.23; N, 4.28.
Found: C, 80.71; H, 5.53; N, 4.27.
Mp 278.0–281.0 °C. 1H NMR (500 MHz, DMSO-d6) d: 7.97 (dd,
2H, J = 5.5, 3.1 Hz), 7.90 (dd, 2H, J = 5.5, 3.1 Hz), 7.67 (d, 2H,
J = 8.5 Hz), 7.40 (d, 2H, J = 8.5 Hz), 7.14 (d, 1H, J = 16.2 Hz), 7.01
(d, 1H, J = 2.4 Hz), 6.97 (d, 1H, J = 16.2 Hz), 6.89 (dd, 1H, J = 8.5,
1.8 Hz), 6.73 (d, 1H, J = 7.9 Hz). FAB-MS m/z: 357 [M]+, 358
[M+H]+. Anal. Calcd for C22H15NO4ꢀ1/4H2O: C, 73.02; H, 4.32; N,
3.87. Found: C, 73.21; H, 4.36; N, 3.91.
4.2.34. N-{4-[2-(3,4-Dimethoxyphenyl)ethyl]phenyl}phthalimide
(21b)
This compound was prepared from 16b by means of a proce-
dure similar to that used for 19b. Compound 21b was obtained
in 79% yield as yellow plates after recrystallization from CH2Cl2/
n-hexane.
4.2.39. N-{4-[2-(3-Methoxyphenyl)ethyl]phenyl}phthalimide
(23d)
This compound was prepared from 16d by means of a proce-
dure similar to that used for 19b. Compound 23d was obtained
in 78% yield as colorless needles after recrystallization from
CH2Cl2/n-hexane.
Mp 180.0–181.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.94 (dd, 2H,
J = 5.5, 3.1 Hz), 7.77 (dd, 2H, J = 5.5, 3.1 Hz), 7.33–7.28 (m, 4H),
6.79 (d, 1H, J = 7.9 Hz), 6.73 (dd, 1H, J = 7.9, 1.8 Hz), 6.63 (d, 1H,
J = 1.8 Hz), 3.85 (s, 3H), 3.82 (s, 3H), 2.95–2.92 (m, 2H), 2.90–2.87
(m, 2H). FAB-MS m/z: 387 [M]+, 388 [M+H]+. Anal. Calcd for
C24H21NO4ꢀ1/5H2O: C, 73.72; H, 5.52; N, 3.58. Found: C, 73.98; H,
5.44; N, 3.64.
Mp 127.0–127.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 3.1 Hz), 7.79 (dd, 2H, J = 5.5, 3.1 Hz), 7.36–7.32 (m, 4H),
7.22 (t, 1H, J = 7.9 Hz), 6.82 (d, 1H, J = 7.3 Hz), 6.76 (dd, 1H,
J = 7.3, 1.8 Hz), 6.74 (d, 1H, J = 1.8 Hz), 3.79 (s, 3H), 3.00–2.96 (m,
2H), 2.95–2.91 (m, 2H). FAB-MS m/z: 357 [M]+, 358 [M+H]+. Anal.
Calcd for C23H19NO3ꢀ1/4H2O: C, 76.33; H, 5.43; N, 3.87. Found: C,
76.34; H, 5.40; N, 3.87.
4.2.35. N-{4-[2-(3,4-Dihydroxyphenyl)ethyl]phenyl}phthalimide
(21c)
This compound was prepared from 21b by means of a proce-
dure similar to that used for 19c. Compound 21c was obtained in
27% yield as yellow needles after recrystallization from CH2Cl2/n-
hexane.
4.2.40. N-{4-[2-(3-Hydroxyphenyl)ethyl]phenyl}phthalimide
(23e)
This compound was prepared from 23d by means of a proce-
dure similar to that used for 19c. Compound 23e was obtained in
68% yield as colorless needles after recrystallization from EtOAc/
n-hexane.
Mp 205.0–206.0 °C. 1H NMR (500 MHz, DMSO-d6) d: 8.73 (br s,
1H), 8.65 (br s, 1H), 7.95 (dd, 2H, J = 5.5, 3.1 Hz), 7.90 (dd, 2H,
J = 5.5, 3.1 Hz), 7.36–7.31 (m, 4H), 6.64 (d, 1H, J = 1.8 Hz), 6.62
(d, 1H, J = 7.9 Hz), 6.49 (dd, 1H, J = 7.9, 1.8 Hz), 2.87–2.84 (m,
2H), 2.75–2.71 (m, 2H). FAB-MS m/z: 359 [M]+, 360 [M+H]+.
HRMS (FAB) calcd for C22H17NO4 359.1158; found: 359.1179
(M)+.
Mp 229.0–232.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 3.1 Hz), 7.80 (dd, 2H, J = 5.5, 3.1 Hz), 7.35–7.30 (m, 4H),
7.17 (t, 1H, J = 7.9 Hz), 6.80 (d, 1H, J = 7.3 Hz), 6.68 (dd, 1H,
J = 7.9, 2.4 Hz), 6.60 (s, 1H), 2.98–2.95 (m, 2H), 2.92–2.89 (m, 2H).
FAB-MS m/z: 343 [M]+, 344 [M+H]+. Anal. Calcd for C22H17NO3ꢀ1/
5H2O: C, 76.15; H, 5.05; N, 4.04. Found: C, 76.38; H, 5.24; N, 4.05.
4.2.36. N-{4-[(1E)-2-Phenylethenyl]phenyl}phthalimide (22a)
This compound was prepared from 18a by means of a procedure
similar to that used for 19b. Compound 22a was obtained in 88%
yield as a white powder after recrystallization from CH2Cl2/n-
hexane.
4.2.41. N-{4-[2-(4-Methoxyphenyl)ethyl]phenyl}phthalimide
(23f)
This compound was prepared from 16f by means of a procedure
similar to that used for 19b. Compound 23f was obtained in 93%
yield as colorless needles after recrystallization from CH2Cl2/n-
hexane.
Mp 297.0–299.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.97 (dd, 2H,
J = 5.5, 3.1 Hz), 7.81 (dd, 2H, J = 5.5, 3.1 Hz), 7.65 (d, 2H,
J = 8.5 Hz), 7.54 (d, 2H, J = 7.9 Hz), 7.46 (d, 2H, J = 8.5 Hz), 7.38
(t, 2H, J = 7.9 Hz), 7.28 (tt, 1H, J = 7.9, 1.2 Hz), 7.15 (s, 2H). FAB-
MS m/z: 325 [M]+, 326 [M+H]+. Anal. Calcd for C22H15NO2ꢀ1/
3H2O: C, 79.74; H, 4.77; N, 4.23. Found: C, 79.50; H, 4.69; N,
4.19.
Mp 196.5–199.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 3.1 Hz), 7.79 (dd, 2H, J = 5.5, 3.1 Hz), 7.36–7.31 (m, 4H),
7.13 (d, 2H, J = 8.5 Hz), 6.85 (d, 2H, J = 9.2 Hz), 3.80 (s, 3H). FAB-
MS m/z: 357 [M]+, 358 [M+H]+. Anal. Calcd for C23H19NO3: C,
77.29; H, 5.36; N, 3.92. Found: C, 77.21; H, 5.51; N, 3.83.

K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
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4.2.42. N-{4-[2-(4-Hydroxyphenyl)ethyl]phenyl}phthalimide
(23g)
[M+5]+. Anal. Calcd for C22H13Cl4NO2: C, 56.81; H, 2.82; N, 3.01.
Found: C, 56.66; H, 2.91; N, 2.95.
This compound was prepared from 23f by means of a procedure
similar to that used for 19c. Compound 23g was obtained in 29%
yield as a brown powder after recrystallization from EtOAc/n-
hexane.
4.2.47. 4,5,6,7-Tetrachloro-N-{3-[2-(3,4-dimethoxyphenyl)ethyl]
phenyl}phthalimide (25b)
This compound was prepared from 15b by means of a proce-
dure similar to that used for 19b. Compound 25b was obtained
in 90% yield as yellow needles after recrystallization from
CH2Cl2/n-hexane.
Mp 285.0–288.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.96 (dd, 2H,
J = 5.5, 3.1 Hz), 7.80 (dd, 2H, J = 5.5, 3.1 Hz), 7.35–7.30 (m, 4H),
7.08 (d, 2H, J = 7.9 Hz), 6.77 (d, 2H, J = 8.5 Hz), 2.94–2.92 (m, 2H),
2.90–2.88 (m, 2H). FAB-MS m/z: 343 [M]+, 344 [M+H]+. Anal. Calcd
for C22H17NO3ꢀ1/2H2O: C, 74.99; H, 5.15; N, 3.97. Found: C, 74.97;
H, 5.20; N, 3.89.
Mp 166.0–167.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.43–7.39 (m,
1H), 7.24–7.20 (m, 3H), 6.79 (d, 1H, J = 7.9 Hz), 6.71 (dd, 1H, J = 7.9,
1.8 Hz), 6.64 (d, 1H, J = 1.8 Hz), 3.86 (s, 3H), 3.83 (s, 3H), 2.97–2.94
(m, 2H), 2.91–2.88 (m, 2H). FAB-MS m/z: 523 [M]+, 524 [M+H]+,
525 [M+2]+, 526 [M+3]+, 527 [M+4]+, 528 [M+5]+. Anal. Calcd for
C24H17Cl4NO4ꢀ1/4H2O: C, 54.42; H, 3.33; N, 2.64. Found: C, 54.49;
H, 3.36; N, 2.61.
4.2.43. 4,5,6,7-Tetrachloro-N-[2-(2-phenylethyl)phenyl]phthali-
mide (24a)
This compound was prepared from 14a by means of a procedure
similar to that used for 19b. Compound 24a was obtained in 82%
yield as colorless plates after recrystallization from CH2Cl2/n-
hexane.
4.2.48. 4,5,6,7-Tetrachloro-N-{3-[2-(3,4-dihydroxyphenyl)ethyl]
phenyl}phthalimide (25c)
Mp 148.0–149.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.42–7.39 (m,
1H), 7.35–7.32 (m, 2H), 7.19–7.16 (m, 2H), 7.14–7.09 (m, 2H), 7.06
(d, 2H, J = 6.7 Hz), 2.87–2.84 (m, 2H), 2.78–2.75 (m, 2H). FAB-MS
m/z: 463 [M]+, 464 [M+H]+, 465 [M+2]+, 466 [M+3]+, 467 [M+4]+,
468 [M+5]+. Anal. Calcd for C22H13Cl4NO2: C, 56.81; H, 2.82; N,
3.01. Found: C, 56.67; H, 2.92; N, 2.94.
This compound was prepared from 25b by means of a proce-
dure similar to that used for 19c. Compound 25c was obtained in
70% yield as a pale yellow powder after recrystallization from
EtOAc/n-hexane.
Mp 208.0–210.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.43 (t, 1H,
J = 7.9 Hz), 7.24 (d, 1H, J = 7.3 Hz), 7.20 (m, 1H), 7.10 (s, 1H), 6.78
(d, 1H, J = 7.9 Hz), 6.63 (dd, 1H, J = 7.9, 1.8 Hz), 6.58 (d, 1H,
J = 1.8 Hz), 5.53 (s, 1H), 5.15 (s, 1H), 2.93–2.91 (m, 2H), 2.85–2.82
(m, 2H). FAB-MS m/z: 495 [M]+, 496 [M+H]+, 497 [M+2]+, 498
[M+3]+, 499 [M+4]+, 500 [M+5]+. Anal. Calcd for C22H13Cl4NO4ꢀ2/
3H2O: C, 51.90; H, 2.84; N, 2.75. Found: C, 51.70; H, 3.03; N, 2.60.
4.2.44. 4,5,6,7-Tetrachloro-N-{2-[2-(3,4-dimethoxyphenyl)ethyl]
phenyl}phthalimide (24b)
This compound was prepared from 14b by means of a proce-
dure similar to that used for 19b. Compound 24b was obtained
in 92% yield as a yellow powder after recrystallization from
CH2Cl2/n-hexane.
4.2.49. 4,5,6,7-Tetrachloro-N-[4-(2-phenylethyl)phenyl]
phthalimide (26a)
Mp 157.0–158.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.41 (td, 1H,
J = 7.9, 1.8 Hz), 7.35 (td, 1H, J = 7.9, 1.8 Hz), 7.32 (dd, 1H, J = 7.3,
1.8 Hz), 7.15 (dd, 1H, J = 7.9, 1.8 Hz), 6.68 (d, 1H, J = 7.9 Hz), 6.61
(dd, 1H, J = 7.9, 1.8 Hz), 6.53 (d, 1H, J = 1.8 Hz), 3.81 (s, 3H), 3.75
(s, 3H), 2.80 (s, 4H). FAB-MS m/z: 523 [M]+, 524 [M+H]+, 525
[M+2]+, 526 [M+3]+, 527 [M+4]+, 528 [M+5]+. Anal. Calcd for
C24H17Cl4NO4: C, 54.88; H, 3.26; N, 2.67. Found: C, 54.63; H,
3.37; N, 2.54.
This compound was prepared from 16a by means of a procedure
similar to that used for 19b. Compound 26a was obtained in 96%
yield as colorless needles after recrystallization from CH2Cl2/n-
hexane.
Mp 222.0–223.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.33–7.28 (m,
6H), 7.22–7.19 (m, 3H), 3.00–2.98 (m, 2H), 2.97–2.94 (m, 2H). FAB-
MS m/z: 463 [M]+, 464 [M+H]+, 465 [M+2]+, 466 [M+3]+, 467
[M+4]+, 468 [M+5]+. Anal. Calcd for C22H13Cl4NO2: C, 56.81; H,
2.82; N, 3.01. Found: C, 56.85; H, 3.02; N, 2.94.
4.2.45. 4,5,6,7-Tetrachloro-N-{2-[2-(3,4-dihydroxyphenyl)ethyl]
phenyl}phthalimide (24c)
This compound was prepared from 24b by means of a proce-
dure similar to that used for 19c. Compound 24c was obtained in
90% yield as colorless plates after recrystallization from EtOAc/n-
hexane.
4.2.50. 4,5,6,7-Tetrachloro-N-{4-[2-(3,4-dimethoxyphenyl)ethyl]
phenyl}phthalimide (26b)
This compound was prepared from 16b by means of a proce-
dure similar to that used for 19b. Compound 26b was obtained
in 99% yield as a yellow powder after recrystallization from
CH2Cl2/n-hexane.
Mp 227.0–229.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.41 (m, 1H),
7.36–7.32 (m, 2H), 7.14–7.13 (m, 1H), 6.66 (d, 1H, J = 7.9 Hz),
6.53 (d, 1H, J = 2.4 Hz), 6.49 (dd, 1H, J = 7.9, 1.8 Hz), 5.09 (s, 1H),
4.94 (s, 1H), 2.76 (s, 4H). FAB-MS m/z: 495 [M]+, 496 [M+H]+, 497
[M+2]+, 498 [M+3]+, 499 [M+4]+, 500 [M+5]+. Anal. Calcd for
C22H13Cl4NO4: C, 53.15; H, 2.64; N, 2.82. Found: C, 53.03; H,
2.88; N, 2.62.
Mp 220.0–221.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.28 (s, 4H),
6.78 (d, 1H, J = 8.5 Hz), 6.71 (dd, 1H, J = 8.5, 1.8 Hz), 6.62 (d, 1H,
J = 1.8 Hz), 3.85 (s, 3H), 3.82 (s, 3H), 2.96–2.93 (m, 2H), 2.90–2.87
(m, 2H). FAB-MS m/z: 523 [M]+, 524 [M+H]+, 525 [M+2]+, 526
[M+3]+, 527 [M+4]+, 528 [M+5]+. HRMS (FAB) calcd for
C24H17Cl4NO4 522.9912; found: 522.9929 (M)+.
4.2.46. 4,5,6,7-Tetrachloro-N-[3-(2-phenylethyl)phenyl]phthali-
mide (25a)
4.2.51. 4,5,6,7-Tetrachloro-N-{4-[2-(3,4-dihydroxyphenyl)ethyl]
phenyl}phthalimide (26c)
This compound was prepared from 15a by means of a procedure
similar to that used for 19b. Compound 25a was obtained in 89%
yield as colorless needles after recrystallization from CH2Cl2/n-
hexane.
This compound was prepared from 26b by means of a proce-
dure similar to that used for 19c. Compound 26c was obtained in
64% yield as a yellow powder after recrystallization from EtOAc/
n-hexane.
Mp 187.0–187.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.42 (t, 1H,
J = 7.9 Hz), 7.29 (t, 2H, J = 7.9 Hz), 7.26–7.25 (m, 1H), 7.24 (dd,
2H, J = 7.9, 1.8 Hz), 7.20 (dd, 1H, J = 7.9, 1.2 Hz), 7.19 (d, 2H,
J = 7.9 Hz), 3.00–2.97 (m, 2H), 2.96–2.94 (m, 2H). FAB-MS m/z:
463 [M]+, 464 [M+H]+, 465 [M+2]+, 466 [M+3]+, 467 [M+4]+, 468
Mp 260.0–263.0 °C. 1H NMR (500 MHz, DMSO-d6) d: 8.70 (s,
1H), 8.62 (s, 1H), 7.38 (d, 2H, J = 8.5 Hz), 7.30 (d, 2H, J = 8.5 Hz),
6.64 (d, 1H, J = 2.1 Hz), 6.62 (d, 1H, J = 7.9 Hz), 6.47 (dd, 1H,
J = 7.9, 2.1 Hz), 2.88–2.85 (m, 2H), 2.75–2.72 (m, 2H). FAB-MS m/

5012
K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
z: 495 [M]+, 496 [M+H]+, 497 [M+2]+, 498 [M+3]+, 499 [M+4]+, 500
[M+5]+. Anal. Calcd for C22H13Cl4NO4ꢀ1/3H2O: C, 52.52; H, 2.74; N,
2.78. Found: C, 52.47; H, 2.85; N, 2.51.
Mp 281.0–284.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.62 (d, 2H,
J = 8.5 Hz), 7.40 (d, 2H, J = 8.5 Hz), 7.11 (d, 1H, J = 16.2 Hz), 7.09–
7.05 (m, 2H), 7.00 (d, 1H, J = 16.2 Hz), 6.88 (d, 1H, J = 8.5 Hz), 3.96
(s, 3H), 3.91 (s, 3H). FAB-MS m/z: 521 [M]+, 522 [M+H]+, 523
[M+2]+, 524 [M+3]+, 525 [M+4]+, 526 [M+5]+. Anal. Calcd for
C24H15Cl4NO4ꢀ1/2H2O: C, 54.16; H, 3.03; N, 2.63. Found: C, 54.24;
H, 2.97; N, 2.55.
4.2.52. 4,5,6,7-Tetrachloro-N-{3-[(1E)-2-phenylethenyl]phenyl}
phthalimide (27a)
This compound was prepared from 17a by means of a procedure
similar to that used for 19b. Compound 27a was obtained in 77%
yield as a white powder after recrystallization from CH2Cl2/n-
hexane.
4.2.57. 4,5,6,7-Tetrachloro-N-{4-[(1E)-2-(3,4-dihydroxyphenyl)
ethenyl]phenyl}phthalimide (28c)
Mp 259.5–261.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.58–7.56 (m,
2H), 7.52–7.49 (m, 3H), 7.37 (t, 2H, J = 7.9 Hz), 7.31–7.28 (m, 2H),
7.15 (d, 1H, J = 16.8 Hz), 7.12 (d, 1H, J = 16.8 Hz). FAB-MS m/z:
461 [M]+, 462 [M+H]+, 463 [M+2]+, 464 [M+3]+, 465 [M+4]+, 466
[M+5]+. Anal. Calcd for C22H11Cl4NO2ꢀ1/3H2O: C, 56.32; H, 2.51;
N, 2.99. Found: C, 56.55; H, 2.56; N, 2.98.
This compound was prepared from 28b by means of a proce-
dure similar to that used for 19c. Compound 28c was obtained in
52% yield as red plates after recrystallization from EtOAc/n-hexane.
Mp >300 °C. 1H NMR (500 MHz, DMSO-d6) d: 7.69 (d, 2H,
J = 8.5 Hz), 7.38 (d, 2H, J = 8.5 Hz), 7.16 (d, 1H, J = 16.5 Hz), 7.02
(d, 1H, J = 1.8 Hz), 6.98 (d, 1H, J = 16.5 Hz), 6.89 (dd, 1H, J = 8.5,
1.8 Hz), 6.73 (d, 1H, J = 7.9 Hz). FAB-MS m/z: 493 [M]+, 494
[M+H]+, 495 [M+2]+, 496 [M+3]+, 497 [M+4]+, 498 [M+5]+. Anal.
Calcd for C22H11Cl4NO4ꢀ1/5H2O: C, 52.98; H, 2.30; N, 2.81. Found:
C, 53.31; H, 2.64; N, 2.64.
4.2.53. 4,5,6,7-Tetrachloro-N-{3-[(1E)-2-(3,4-dimethoxyphenyl)
ethenyl]phenyl}phthalimide (27b)
This compound was prepared from 17b by means of a proce-
dure similar to that used for 19b. Compound 27b was obtained
in 73% yield as a yellow powder after recrystallization from
CH2Cl2/n-hexane.
4.2.58. 4,5,6,7-Tetrachloro-N-{3-[2-(3-methoxyphenyl)ethyl]
phenyl}phthalimide (29d)
Mp 248.0–249.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.56–7.54 (m,
2H), 7.49 (t, 1H, J = 7.9 Hz), 7.29–7.27 (m, 1H), 7.09 (d, 1H,
J = 16.2 Hz), 7.07–7.05 (m, 2H), 6.99 (d, 1H, J = 16.2 Hz), 6.87 (d,
1H, J = 7.9 Hz), 3.95 (s, 3H), 3.91 (s, 3H). FAB-MS m/z: 521 [M]+,
522 [M+H]+, 523 [M+2]+, 524 [M+3]+, 525 [M+4]+, 526 [M+5]+. Anal.
Calcd for C24H15Cl4NO4ꢀ1/2H2O: C, 54.16; H, 3.03; N, 2.63. Found: C,
53.98; H, 3.03; N, 2.49.
This compound was prepared from 15d by means of a proce-
dure similar to that used for 19b. Compound 29d was obtained
in 83% yield as colorless needles after recrystallization from
CH2Cl2/n-hexane.
Mp 150.0–151.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.42 (t, 1H,
J = 7.9 Hz), 7.26–7.25 (m, 2H), 7.23 (m, 1H), 7.20 (t, 1H,
J = 7.9 Hz), 6.78 (d, 1H, J = 7.3 Hz), 6.75 (dd, 1H, J = 7.9, 1.8 Hz),
6.72–6.72 (m, 1H), 3.78 (s, 3H), 3.00–2.96 (m, 2H), 2.94–2.91 (m,
2H). FAB-MS m/z: 493 [M]+, 494 [M+H]+, 495 [M+2]+, 496 [M+3]+,
497 [M+4]+, 498 [M+5]+. Anal. Calcd for C23H15Cl4NO3ꢀ1/4H2O: C,
55.28; H, 3.13; N, 2.80. Found: C, 55.43; H, 3.11; N, 2.82.
4.2.54. 4,5,6,7-Tetrachloro-N-{3-[(1E)-2-(3,4-dihydroxyphenyl)
ethenyl]phenyl}phthalimide (27c)
This compound was prepared from 27b by means of a proce-
dure similar to that used for 19c. Compound 27c was obtained in
92% yield as orange needles after recrystallization from EtOAc/n-
hexane.
4.2.59. 4,5,6,7-Tetrachloro-N-{3-[2-(3-hydroxyphenyl)ethyl]
phenyl}phthalimide (29e)
Mp 257.0–260.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.51 (dt, 1H,
J = 7.9, 1.2 Hz), 7.50 (m, 1H), 7.47 (t, 1H, J = 7.9 Hz), 7.26–7.25
(m, 1H), 7.05 (d, 1H, J = 1.8 Hz), 7.00 (d, 1H, J = 16.5 Hz), 6.95
(dd, 1H, J = 8.5, 1.8 Hz), 6.91 (d, 1H, J = 16.5 Hz), 6.85 (d, 1H,
J = 8.5 Hz), 5.59 (br s, 1H), 5.54 (br s, 1H). FAB-MS m/z: 493
[M]+, 494 [M+H]+, 495 [M+2]+, 496 [M+3]+, 497 [M+4]+, 498
[M+5]+. HRMS (FAB) calcd for C22H11Cl4NO4 492.9442; found:
492.9467 (M)+.
This compound was prepared from 29d by means of a proce-
dure similar to that used for 19c. Compound 29e was obtained in
60% yield as colorless needles after recrystallization from EtOAc/
n-hexane.
Mp 213.0–215.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.42 (t, 1H,
J = 7.9 Hz), 7.25–7.20 (m, 3H), 7.15 (t, 1H, J = 7.9 Hz), 6.76 (d, 1H,
J = 7.9 Hz), 6.68 (dd, 1H, J = 7.9, 2.4 Hz), 6.62 (t, 1H, J = 1.8 Hz),
2.98–2.95 (m, 2H), 2.92–2.88 (m, 2H). FAB-MS m/z: 479 [M]+,
480 [M+H]+, 481 [M+2]+, 482 [M+3]+, 483 [M+4]+, 484 [M+5]+. Anal.
Calcd for C22H13Cl4NO3: C, 54.92; H, 2.72; N, 2.91. Found: C, 54.68;
H, 2.99; N, 2.77.
4.2.55. 4,5,6,7-Tetrachloro-N-{4-[(1E)-2-phenylethenyl]phenyl}
phthalimide (28a)
This compound was prepared from 18a by means of a procedure
similar to that used for 19b. Compound 28a was obtained in 73%
yield as yellow needles after recrystallization from CH2Cl2/n-
hexane.
4.2.60. 4,5,6,7-Tetrachloro-N-{3-[2-(4-methoxyphenyl)ethyl]
phenyl}phthalimide (29f)
This compound was prepared from 15f by means of a procedure
similar to that used for 19b. Compound 29f was obtained in 74%
yield as pale yellow plates after recrystallization from CH2Cl2/n-
hexane.
Mp 296.0–296.5 °C. 1H NMR (500 MHz, CDCl3) d: 7.65 (d, 2H,
J = 8.5 Hz), 7.54 (d, 2H, J = 7.3 Hz), 7.42 (d, 2H, J = 8.5 Hz), 7.38 (t,
2H, J = 7.3 Hz), 7.29 (tt, 1H, J = 7.3, 1.2 Hz), 7.17 (d, 1H,
J = 16.5 Hz), 7.13 (d, 1H, J = 16.5 Hz). FAB-MS m/z: 461 [M]+, 462
[M+H]+, 463 [M+2]+, 464 [M+3]+, 465 [M+4]+, 466 [M+5]+. Anal.
Calcd for C22H11Cl4NO2: C, 57.05; H, 2.39; N, 3.02. Found: C,
56.83; H, 2.69; N, 3.09.
Mp 169.0–170.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.41 (t, 1H,
J = 8.5 Hz), 7.23–7.22 (m, 2H), 7.09 (d, 2H, J = 8.5 Hz), 6.83 (d, 2H,
J = 8.5 Hz), 3.79 (s, 3H), 2.96–2.93 (m, 2H), 2.91–2.87 (m, 2H).
FAB-MS m/z: 494 [M+H]+, 495 [M+2]+, 496 [M+3]+, 497 [M+4]+,
498 [M+5]+. Anal. Calcd for C23H15Cl4NO3ꢀ1/4H2O: C, 55.28; H,
3.13; N, 2.80. Found: C, 55.28; H, 3.09; N, 2.78.
4.2.56. 4,5,6,7-Tetrachloro-N-{4-[(1E)-2-(3,4-dimethoxyphenyl)
ethenyl]phenyl}phthalimide (28b)
This compound was prepared from 18b by means of a proce-
dure similar to that used for 19b. Compound 28b was obtained
in 68% yield as an orange powder after recrystallization from
CH2Cl2/n-hexane.
4.2.61. 4,5,6,7-Tetrachloro-N-{3-[2-(4-hydroxyphenyl)ethyl]
phenyl}phthalimide (29g)
This compound was prepared from 29f by means of a procedure
similar to that used for 19c. Compound 29g was obtained in 67%

K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
5013
yield as a white powder after recrystallization from EtOAc/n-
hexane.
tion, Culture, Sports, Science and Technology, Japan, and the Japan
Society for the Promotion of Science.
Mp 210.0–212.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.41 (t, 1H,
J = 7.3 Hz), 7.24–7.20 (m, 3H), 7.03 (d, 2H, J = 8.5 Hz), 6.75 (d, 2H,
J = 8.5 Hz), 4.60 (br s, 1H), 2.95–2.92 (m, 2H), 2.89–2.86 (m, 2H).
FAB-MS m/z: 479 [M]+, 480 [M+H]+, 481 [M+2]+, 482 [M+3]+, 483
[M+4]+, 484 [M+5]+. Anal. Calcd for C22H13Cl4NO3: C, 54.92; H,
2.72; N, 2.91. Found: C, 54.68; H, 2.82; N, 2.83.
References and notes
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Å. Proc. Natl. Acad. Sci. U.S.A. 1995, 92, 2096.
4. Glass, C. K.; Rosenfeld, M. G. Genes Dev. 2000, 14, 121.
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4.2.62. 4,5,6,7-Tetrachloro-N-{4-[2-(3-methoxyphenyl)ethyl]
phenyl}phthalimide (30d)
This compound was prepared from 16d by means of a proce-
dure similar to that used for 19b. Compound 30d was obtained
in 95% yield as colorless plates after recrystallization from
CH2Cl2/n-hexane.
7. Peet, D. J.; Turley, S. D.; Ma, W.; Janowski, B. A.; Lobaccaro, J.-M. A.; Hammer, R.
E.; Mangelsdorf, D. J. Cell 1998, 93, 693.
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Heyman, R. A.; Dietschy, J. M.; Mangelsdorf, D. J. Science 2000, 289, 1524.
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J. J. Biol. Chem. 2002, 277, 18793.
11. Kennedy, M. A.; Venkateswaran, A.; Tarr, P. T.; Xenarious, I.; Kudoh, J.; Shimizu,
N.; Edwards, P. A. J. Biol. Chem. 2001, 276, 39438.
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13. Terasaka, N.; Hiroshima, A.; Koieyama, T.; Ubukata, N.; Morikawa, Y.; Nakai, D.;
Inaba, T. FEBS Lett. 2003, 536, 6.
Mp 170.5–172.0 °C. 1H NMR (500 MHz, CDCl3) d: 7.34–7.29 (m,
4H), 7.22 (t, 1H, J = 7.9 Hz), 6.80 (d, 1H, J = 7.3 Hz), 6.76 (dd, 1H,
J = 7.9, 1.8 Hz), 6.73 (s, 1H), 3.79 (s, 3H), 3.00–2.97 (m, 2H), 2.95–
2.91 (m, 2H). FAB-MS m/z: 493 [M]+, 494 [M+H]+, 495 [M+2]+,
496 [M+3]+, 497 [M+4]+, 498 [M+5]+. HRMS (FAB) calcd for
C23H15Cl4NO3 492.9806; found: 492.9834 (M)+.
14. Joseph, S. B.; McKilligin, E.; Pei, L.; Watson, M. A.; Collins, A. R.; Laffitte, B. A.;
Chen, M.; Noh, G.; Goodman, J.; Hagger, G. N.; Tran, J.; Tippin, T. K.; Wang, X.;
Lusis, A. J.; Hsueh, W. A.; Law, R. E.; Collins, J. L.; Willson, T. M.; Tontonoz, P.
Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 7604.
4.2.63. 4,5,6,7-Tetrachloro-N-{4-[2-(3-hydroxyphenyl)ethyl]
phenyl}phthalimide (30e)
15. Schultz, J. R.; Tu, H.; Luk, A.; Repa, J. J.; Medina, J. C.; Li, L.; Schwendner, S.;
Wang, S.; Thoolen, M.; Mangelsdorf, D. J.; Lustig, K. D.; Shan, B. Genes Dev. 2000,
14, 2831.
16. Collins, J. L.; Fivush, A. M.; Watson, M. A.; Galardi, C. M.; Lewis, M. C.; Moore, L.
B.; Parks, D. J.; Wilson, J. G.; Tippin, T. K.; Binz, J. G.; Plunket, K. D.; Morgan, D.
G.; Beaudet, E. J.; Whitney, K. D.; Kliewer, S. A.; Willson, T. M. J. Med. Chem.
2002, 45, 1963.
17. Joseph, S. B.; Laffitte, B. A.; Patel, P. H.; Watson, M. A.; Matsukuma, K. E.;
Walczak, R.; Collins, J. L.; Osborne, T. F.; Tontonoz, P. J. Biol. Chem. 2002, 277,
11019.
18. Lund, E. G.; Peterson, L. B.; Adams, A. D.; Lam, M.-H. N.; Burton, C. A.; Chin, J.;
Guo, Q.; Huang, S.; Latham, M.; Lopez, J. C.; Menke, J. G.; Milot, D. P.; Mitnaul, L.
J.; Rex-Rabe, S. E.; Rosa, R. L.; Tian, J. Y.; Wright, S. D.; Sparrow, C. P. Biochem.
Pharmacol. 2006, 71, 453.
This compound was prepared from 30d by means of a proce-
dure similar to that used for 19c. Compound 30e was obtained in
66% yield as a pale brown powder after recrystallization from
EtOAc/n-hexane.
Mp 267.0–268.0 °C. 1H NMR (500 MHz, DMSO-d6) d: 9.25 (br s,
1H), 7.40 (d, 2H, J = 7.9 Hz), 7.31 (d, 2H, J = 7.9 Hz), 7.06 (t, 1H,
J = 7.3 Hz), 6.67 (d, 1H, J = 7.3 Hz), 6.66 (s, 1H), 6.58 (d, 1H,
J = 7.3 Hz), 2.92–2.90 (m, 2H), 2.84–2.81 (m, 2H). FAB-MS m/z:
479 [M]+, 480 [M+H]+, 481 [M+2]+, 482 [M+3]+, 483 [M+4]+, 484
[M+5]+. HRMS (FAB) calcd for C22H13Cl4NO3 478.9650; found:
478.9694 (M)+.
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Acknowledgments
The work described in this paper was partially supported by
Grants-in-Aid for Scientific Research from The Ministry of Educa-

5014
K. Motoshima et al. / Bioorg. Med. Chem. 17 (2009) 5001–5014
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