Discovery of an orally-active nonsteroidal androgen receptor pure antagonist and the structure-activity relationships of its derivatives

Article abstract of DOI:10.1248/cpb.56.1555

The 3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethylthiohydantoin derivatives which have carboxy-terminal side chains were synthesized and their agonistic/antagonistic activities against androgen receptor (AR) measured. Among them, compound 13b showed antagonistic activity (IC₅₀=130 nM) with no agonistic activity even at 10000 nM. This compound exhibited significant metabolic stability and oral antiandrogenic activity (ED₅₀=7 mg/kg).

Full text of DOI:10.1248/cpb.56.1555

November 2008
Chem. Pharm. Bull. 56(11) 1555—1561 (2008)
1555
Discovery of an Orally-Active Nonsteroidal Androgen Receptor Pure
Antagonist and the Structure–Activity Relationships of Its Derivatives
Kazutaka TACHIBANA,* Ikuhiro IMAOKA, Takuya SHIRAISHI, Hitoshi YOSHINO, Mitsuaki NAKAMURA,
Masateru OHTA, Hiromitsu KAWATA, Kenji TANIGUCHI, Nobuyuki ISHIKURA, Toshiaki TSUNENARI,
Hidemi SAITO, Masahiro NAGAMUTA, Toshito NAKAGAWA, Kenji TAKANASHI, Etsuro ONUMA, and
Haruhiko SATO
Fuji Gotemba Research Laboratories, Chugai Pharmaceutical Co., Ltd.; 1–135 Komakado, Gotemba, Shizuoka 412–8513,
Japan. Received June 14, 2008; accepted September 5, 2008
The 3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethylthiohydantoin derivatives which have carboxy-termi-
nal side chains were synthesized and their agonistic/antagonistic activities against androgen receptor (AR) meas-
ured. Among them, compound 13b showed antagonistic activity (IC50ꢀ130 nM) with no agonistic activity even at
10000 nM. This compound exhibited significant metabolic stability and oral antiandrogenic activity (ED50ꢀ
7 mg/kg).
Key words androgen receptor; pure antagonist; prostate cancer
Prostate cancer is the most common cancer amongst men antagonists. However, our compounds didn’t show antiandro-
and the second most common malignant cause of male death genic activity in vivo, probably due to the lack of metabolic
after lung cancer in the U.S.1) Since the growth of prostate stability of the steroidal core structures.12)
cancer is dependent on androgen, androgen receptor (AR)
Among a number of nonsteroidal compounds that have
antagonists such as flutamide (1), a precursor of its active been reported to have binding affinities for AR, thiohydan-
form hydroxyflutamide (2), and bicalutamide (3) are cur- toin derivative RU56187 (5) has a high affinity for AR and
rently used as hormone therapy (Chart 1).2) These antiandro- oral antiandrogenic activity in vivo (Chart 1).14) But this
gens exhibit good efficacy in many cases and comprise an compound also showed agonistic activities in RGA.15,16) Cou-
important part of effective therapeutics.3—6) However, a con- pled with our previous results, we speculated that these ago-
siderable problem with these antiandrogens is that recurrence nistic activities are caused by the lack of a side chain to in-
occurs after a short period of response.7) They have partial hibit the folding of helix 12. The nitrogen atom on position 1
agonistic activities at high concentration in vitro,8) which in thiohydantoin seemed appropriate for the introduction of a
may attribute to recurrence. Therefore, the search for new an- side chain and we therefore hypothesized that our strategy
tiandrogenic agents that exhibit no agonistic activities, so- could be applied to thiohydantoin to discover an orally active
called “AR pure antagonists,” has been conducted.9,10)
We previously reported that some steroidal compounds
nonsteroidal AR pure antagonist.
In our previous report, terminal-carboxylic acid side
that have a side chain at the position 7a such as compound 4, chains were introduced to the steroidal core structures be-
exhibited AR pure antagonistic activities in reporter gene cause the carboxylic acid was expected to increase the water-
assay (RGA) (Chart 1).11,12) Since the steroidal core struc- solubility and oral absorption of the compounds.12,17) Unfor-
tures have intrinsic agonistic activities, the side chains are tunately, as mentioned above, the compounds did not show in
critical for pure antagonistic activities. Drawing on the anal- vivo antiandrogenic activity, probably due to the lack of
ogy between the estrogen receptor (ER), for which the mo- metabolic stability of the steroidal core structure. However,
lecular mechanisms of the expression of antagonistic activi- some compounds exhibited AR pure antagonistic activities in
ties have been investigated,13) and AR, the side chain should RGA, suggesting that the terminal-carboxylic acid side
inhibit the folding of helix 12 of the ligand binding domain chains are valuable for the discovery of AR pure antagonists.
(LBD), which is necessary for agonistic activity. Thus, we Furthermore, it was found that the length or the structure of
considered the introduction of a side chain to an AR affinity the linker part of the side chain significantly affects the pure
ligand to be a useful strategy for the discovery of AR pure antagonistic activities. Therefore, we decided to introduce
terminal-carboxylic acid side chains with various types of
linkers to the thiohydantoin core structure in order to clarify
the structure–activity relationships and discover orally active
AR pure antagonists.
We here report the synthesis of novel thiohydantoin deriv-
atives and their structure–activity relationships for AR pure
antagonistic activities. Moreover, we report that one of the
derivatives showed a high metabolic stability in vitro and an-
tiandrogenic activity from oral administration in vivo.
Results and Discussion
The target compounds 13a—h and 18a—d were prepared
Chart 1. Structures of AR Antagonists
according to the synthetic sequences outlined in Charts 2—4.
∗ To whom correspondence should be addressed. e-mail: tachibanakzt@chugai-pharm.co.jp
© 2008 Pharmaceutical Society of Japan

1556
Vol. 56, No. 11
Reagents and conditions: (a) NEt3 or MeOH, r.t.
Chart 2
Reagents and conditions: (a) thiophosgene, THF–H2O; (b) compounds 8a—h, NEt3, THF, reflux; (c) 2 N-HCl, EtOH, reflux; (d) 2 N-NaOH, EtOH, r.t.
Chart 3
Reagents and conditions: (a) NEt3, THF, reflux; (b) 6 N-HCl, dioxane, 80 °C; (c) BBr3, CH2Cl2, ꢁ78 to 0 °C; (d) Br(CH2)mCOOR, NaH, DMF, 0 °C to r.t.; (e) 2 N-NaOH, MeOH,
r.t.
Chart 4
In the preparation of aminonitriles 8a—l by the coupling of which the transcriptional activity is 5% of the transcriptional
corresponding amines 7 with acetone cyanohydrin 6, NEt3 activity of 0.1 nM of DHT. The maximum transcriptional ac-
was used when the amine was HCl salt (Chart 2). Isothio- tivity of each compound indicated as its efficacy (% of 0.1 nM
cyanate 10, which was derived from 9 and thiophosgene, was of DHT). A “pure antagonist” was defined as having an EC5
coupled with compounds 8a—l in THF in the presence of value greater than 10000 nM.
NEt3 under reflux to give 5-imino-2-thioxoimidazolidines
First, we evaluated RU56187 (5) for its binding and tran-
11a—h and 14a—d. The imino groups and terminal esters of scriptional activities for AR in our assay systems to estimate
compounds 11a—h were hydrolyzed successively to give the validity of the compound as a template (Table 1, Fig. 1).
methylene linker compounds 13a—h (Chart 3). Compound As shown in Fig. 1, it exhibited remarkable agonistic activity
14a, in which the protecting group of phenolic OH was the at concentration as low as 1 nM. Although it tended to show
methoxymethyl group, was converted to phenol-terminal antagonistic activities at lower concentrations, its own ago-
thiohydantoin 16a directly by heating in 6 N-HCl and diox- nistic activities diminished the effect at higher concentra-
ane. Compounds 14b—d, in which the hydroxyl groups were tions, thus strongly suggesting that this compound is an ap-
protected by the methyl group, were converted to compounds propriate template for the introduction of a side chain to ver-
15a—c under the same conditions followed by deprotection ify our hypothesis.
of the methyl group by BBr3. Alkylation of the hydroxyl
groups of 16a—d followed by hydrolysis of the terminal es- with a terminal-carboxylic acid side chain are shown in Table
ters gave phenyl-inserted derivatives 18a—d (Chart 4). 1. The activities of the compounds with a simple alkyl side
The agonistic/antagonistic activities of the compounds
The synthesized compounds were evaluated for their in chain (13a—h) were found to be influenced by the length of
vitro binding affinities and agonist/antagonist activities for the alkyl chain. Compound 13a, with two methylene groups,
AR using the same procedures as reported previously.11) The did not show any agonistic or antagonistic activities, possibly
binding affinity for AR was determined by displacement of because of an electric repulsion of a carboxyl group in close
[3H]-mibolerone with the test compound utilizing CHO- proximity to the hydrophobic ligand binding pocket in the
K1/hAR cells. The agonistic and antagonistic activities of the LBD of AR. Compounds with three to five methylene groups
compounds for AR were determined by RGA using hAR- (13b—d) exhibiting antagonistic activities of IC50s of 130—
transfected Hela cells. Antagonistic activity was determined 800 nM, with no agonistic activities even at 10000 nM, were
by the IC50 value, the concentration of a compound that in- best suited to be pure antagonists. Among them, 13b showed
hibits the transcriptional activity of 0.1 nM of DHT by 50%. the highest antagonistic activity (IC50ꢀ130 nM). Compounds
To determine agonistic activity, we calculated the value of with more than 5 methylene groups in the side chain tended
EC5, the concentration of a compound-treated group in to show partial agonistic activities (13e—h). The difference

November 2008
1557
Table 1. Binding and Agonistic/Antagonistic Activitiesa) of Thiohydantoin Derivatives
RGA
Binding
Compound
R
IC50
(nM)
EC5 (nM)
efficacy (%)
IC50 (nM)
RU56187 (5)
13a
Me
15
ꢂ1 (135)
ꢃ10000 (0.5)
ꢃ10000 (2.0)
ꢃ10000 (3.3)
ꢃ10000 (3.1)
2700 (9.6)
NDb)
ꢃ10000
130
–(CH2)2CO2H
–(CH2)3CO2H
–(CH2)4CO2H
–(CH2)5CO2H
–(CH2)6CO2H
–(CH2)7CO2H
–(CH2)8CO2H
–(CH2)9CO2H
ꢃ10000
3900
3800
ꢃ10000
6000
460
13b
13c
13d
13e
13f
13g
13h
330
800
720
380
200
89
3500 (7.9)
4500 (6.6)
3000 (8.8)
180
150
18a
180
370 (28)
160
18b
18c
59
110 (26)
11 (33)
240
60
100
18d
35
54 (16)
45
a) All data are mean values of duplicate experiments. b) ND: Not determined.
Fig. 1. Agonistic and Antagonistic Activities of RU56187 (5) and Compound 13b in Reporter Gene Assay with hAR-Transfected Hela Cells
(A) Dose-dependent agonistic activities of compounds without DHT. (B) Dose-dependent antagonistic activities of compounds in the presence of 0.1 nM of DHT. All data are
mean values of duplicate experiments.
in pure antagonistic activities between 13b and 13e was in- tively high agonistic activities with EC5s of 370 nM and
vestigated using the docking model illustrated in Fig. 2. In 54 nM, respectively. We also investigated the activities of
this model, the side-chain carboxyl group of 13b locates at a compounds 18b and 18c, in which the substituted position of
region in which the hydrophobic helix 12 would be located in the terminal carboxypropyloxy groups were changed. How-
the agonistic form, leading to the inhibition of the folding of ever, these compounds also exhibited only partial agonistic
helix 12. In the case of 13e, the interaction of the terminal activities. The differences in the pure antagonistic activities
carboxyl group with His874 would lead the side chain in a between 4 and 18a were also investigated by a docking
direction allowing the folding of helix 12.
model (Fig. 3). The side-chain carboxyl group of 4 locates at
We also investigated the effects of a side chain with a a region in which hydrophobic helix 12 would be found in
phenyl ring in the center. Previously, we reported that the the agonistic form, resulting in pure antagonistic activity. In
side chain with a 4-(3-carboxypropoxy)phenylpropyl group the case of 18a, the interaction of the terminal carboxyl
and a 3-(3-carboxypropoxy)phenylbutyl group exhibited pure group with the imidazole of His874 and the NH group of the
antagonistic activities when introduced at position 7a of main chain of Leu741 would lead the side chain in a direc-
17a-methyltestosterone.12) However, compounds 18a and tion which allows the folding of helix 12.
18d, which have these groups as a side chain, showed rela-
The metabolic stability of the two thiohydantoin deriva-

1558
Vol. 56, No. 11
Fig. 2. Docking Model of Compounds 13b (Blue) and 13e (Magenta) to
AR (Gray)
Fig. 4. Antiandrogenic Activities on Seminal Vesicle (SV) Wet Weights in
Mice
Each bar represents the meanꢅS.E.M. (nꢀ4—5 animals per group). ∗ pꢂ0.05 com-
pared with the TP group by Dunnett’s multiple-comparison test. # pꢂ0.05 compared
with the TP group by Student’s t test.
to affect pure antagonistic activity. Compound 13b, one of
the pure antagonists found in this investigation, exhibited
substantially high in vitro metabolic stabilities compared to a
steroidal compound. In addition, 13b showed obvious antian-
drogenic activities with an ED50 of 7 mg/kg and a level of ef-
ficacy similar to castration.
Experimental
Column chromatography was carried out on Merck Silicagel 60 (230—
400 mesh) if not otherwise specified. Rf was determined using Merck Sil-
1
icagel 60 F254 plates. H-NMR spectra were recorded on JEOL EX-270 or
JEOL ECP-400. Mass spectra (MS) were measured by Thermo Electron
LCQ Classic (ESI) or Shimadzu GCMS-QP5050A (EI). High resonance
mass spectra (HR-MS) were recorded by a Micromass Q-Tof Ultima API
mass spectrometer or Applied Biosystems QSTAR XL MS/MS system.
Typical Procedures for Synthesis of Carboxyalkylthiohydantoins
(13a—h). 4-[(Cyanodimethylmethyl)amino]butyric Acid Ethyl Ester
(8b) The mixture of acetone cyanohydrin (6) (1.0 g, 11.8 mmol) and 4-
aminobutyric acid ethyl ester HCl salt (7b) (1.97 g, 11.8 mmol) in NEt3
(6 ml, 43.0 mmol) was stirred at room temperature for 1 h. Water was added
and the mixture was extracted with ether. The organic layer was washed with
brine, dried over MgSO4, filtered, and concentrated under vacuum to give
crude compound 8b (1.91 g, 82%) as an oil. This compound was used in the
next step without purification. Rf 0.38 (AcOEt/Hexaneꢀ1 : 2); 1H-NMR
(270 MHz, CDCl3) d: 1.26 (3H, t, Jꢀ7.1 Hz), 1.45 (6H, s), 1.78—1.88 (2H,
m), 2.40 (2H, t, Jꢀ7.3 Hz), 2.76 (2H, t, Jꢀ6.5 Hz), 4.14 (2H, q, Jꢀ7.1 Hz).
4-[3-(4-Cyano-3-trifluoromethylphenyl)-4-imino-5,5-dimethyl-2-thiox-
oimidazolidin-1-yl]butyric Acid Ethyl Ester (11b) To a solution of 4-
isothiocyanato-2-trifluoromethylbenzonitrile (10) (1.2 g, 5.26 mmol) and
compound 8b (1.1 g, 5.37 mmol) in THF (20 ml) was added NEt3 (0.2 ml,
1.43 mmol) and the mixture was stirred at reflux for 50 min. After cooling to
room temperature, the mixture was concentrated under vacuum and purified
by silica gel column chromatography (AcOEt/Hexaneꢀ1 : 1) to give com-
Fig. 3. Docking Model of Compounds 4 (Blue) and 18a (Magenta) to AR
(Gray)
Table 2. Metabolic Stabilities of Compounds in Mouse-Liver Microsomes
CLint
Compound
(ml/min/mg protein)
13b
18a
4
11.5
10.3
219
tives was examined in mouse liver microsomes (Table 2).
Compound 13b, the most potent AR pure antagonist in this
study, showed significantly high metabolic stability. Further-
more, compound 18a also exhibited high metabolic stability
compared to that of compound 4, which is a steroidal deriva- pound 11b (938 mg, 42%) as an oil. Rf 0.28 (AcOEt/hexaneꢀ2 : 1); 1H-
NMR (270 MHz, CDCl3) d: 1.29 (3H, t, Jꢀ7.2 Hz), 1.59 (6H, s), 2.05—2.21
(2H, m), 2.44 (2H, t, Jꢀ6.8 Hz), 3.69—3.75 (2H, m), 4.17 (2H, q,
Jꢀ7.2 Hz), 7.44 (1H, s), 7.64—8.03 (3H, m).
tive with a side chain identical to that of 18a. These results
suggest that the thiohydantoin core structure compared to the
steroidal core structure contributes significantly to the meta-
bolic stability.
Last, the antiandrogenic activities of 13b were evaluated
on seminal vesicle (SV) wet weights in castrated mice (Fig.
4). Compound 13b inhibited seminal vesicle wet-weight gain
by 10 mg/body subcutaneously (s.c.) of testosterone propi-
onate (TP) dose dependently (ED50ꢀ7 mg/kg). Furthermore,
at a dose of 50 mg/kg, the activity reached almost the same
level as with castration.
4-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thiox-
oimidazolidin-1-yl]butyric Acid Ethyl Ester (12b) To a solution of com-
pound 11b (938 mg, 2.2 mmol) in EtOH (10 ml) was added 2 N-HCl (10 ml)
and the mixture was stirred at reflux for 1 h. After cooling to room tempera-
ture, the mixture was extracted with CHCl3. The organic layer was washed
with H2O and brine, dried over Na2SO4, filtered, and concentrated under vac-
uum. The purification by silica gel column chromatography (MeOH/
CHCl3ꢀ0 : 1—1 : 30) gave compound 12b (765 mg, 81%) as an oil. Rf 0.70
(AcOEt/hexaneꢀ2 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.29 (3H, t,
Jꢀ7.1 Hz), 1.62 (6H, s), 2.11—2.20 (2H, m), 2.45 (2H, t, Jꢀ6.7 Hz), 3.73—
3.79 (2H, m), 4.17 (2H, q, Jꢀ7.1 Hz), 7.76 (1H, d, Jꢀ8.3 Hz), 7.89 (1H, s),
7.95 (1H, d, Jꢀ8.3 Hz); MS (ESI) m/z: 428 [(MꢄH)ꢄ].
In summary, we have discovered novel thiohydantoin de-
rivatives which have a carboxy-terminal side chain similar to
AR pure antagonists. The structure of a side chain was found
4-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thiox-
oimidazolidin-1-yl]butyric Acid (13b) To a solution of compound 12b

November 2008
1559
crude products were dissolved in THF (2 ml) and NEt3 (207 ml, 1.5 mmol)
was added and the mixture was stirred at reflux for 1 h. After cooling to
room temperature, water was added and the mixture was extracted with
AcOEt. The organic layer was washed with brine, dried over MgSO4, fil-
tered, and concentrated under vacuum. The purification by silica gel column
chromatography (AcOEt/hexaneꢀ1 : 1) gave compound 14a (442 mg, 43%)
as a colorless oil. Rf 0.33 (AcOEt/Hexaneꢀ1 : 1); 1H-NMR (270 MHz,
CDCl3) d: 1.57 (6H, s), 2.03—2.19 (2H, m), 2.67 (2H, t, Jꢀ7.6 Hz), 3.48
(3H, s), 3.62—3.68 (2H, m), 5.16 (2H, s), 6.98 (2H, d, Jꢀ8.6 Hz), 7.14 (2H,
d, Jꢀ8.6 Hz), 7.62—8.01 (3H, m).
4-{5-Imino-3-[3-(3-methoxyphenyl)propyl]-4,4-dimethyl-2-thioxoimi-
dazolidin-1-yl}-2-trifluoromethylbenzonitrile (14b) This compound was
prepared by a procedure similar to that described for 14a. Colorless oil. Rf
0.23 (AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.58 (6H, s),
2.11—2.22 (2H, m), 2.71 (2H, t, Jꢀ7.5 Hz), 3.63—3.69 (2H, m), 3.81 (3H,
s), 6.75—6.83 (3H, m), 7.23 (1H, t, Jꢀ9.1 Hz), 7.61—8.02 (3H, m).
4-{3-[3-(3-Methoxyphenyl)propyl]-4,4-dimethyl-5-oxo-2-thioxoimida-
zolidin-1-yl}-2-trifluoromethylbenzonitrile (15a) Compound 14b (328
mg, 0.712 mmol) was dissolved in 1,4-dioxane (1 ml) and 6 N-HCl (3 ml) and
the mixture was stirred at 80 °C for 1 h. After cooling to room temperature,
water was added and the mixture was extracted with CHCl3. The organic
layer was washed with brine, dried over MgSO4, filtered, and concentrated
under vacuum. The purification by silica gel column chromatography
(AcOEt/hexaneꢀ1:2) gave compound 15a (98 mg, 30%) as colorless oil. Rf
0.70 (AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.52 (6H, s),
2.15—2.21 (2H, m), 2.71 (2H, t, Jꢀ7.4 Hz), 3.66—3.72 (2H, m), 3.81 (3H,
s), 6.78—6.83 (3H, m), 7.23 (1H, t, Jꢀ8.6 Hz), 7.76 (1H, dd, Jꢀ1.7,
8.3 Hz), 7.88 (1H, d, Jꢀ1.7 Hz), 7.94 (1H, d, Jꢀ8.3 Hz); MS (EI) m/z: 461
[Mꢄ].
(765 mg, 1.8 mmol) in EtOH (8 ml) was added 2 N-NaOH (8 ml) and the
mixture was stirred at room temperature for 1.5 h. After addition of 2 N-HCl
the mixture was extracted with AcOEt. The organic layer was washed with
brine, dried over Na2SO4, filtered, and concentrated under vacuum. The pu-
rification by silica gel column chromatography (MeOH/CHCl3ꢀ1 : 100)
gave compound 13b (445 mg, 64%) as a white solid. mp 161—162 °C
(AcOEt–hexane); Rf 0.13 (AcOEt/Hexaneꢀ1 : 1); 1H-NMR (270 MHz,
CDCl3) d: 1.61 (6H, s), 2.14—2.20 (2H, m), 2.54 (2H, t, Jꢀ6.8 Hz), 3.75—
3.81 (2H, m), 7.77 (1H, dd, Jꢀ1.7, 8.1 Hz), 7.89 (1H, d, Jꢀ1.7 Hz), 7.95
(1H, d, Jꢀ8.1 Hz); MS (ESI) m/z: 400 [(MꢄH)ꢄ]; Anal. Calcd for
C17H16F3N3O3S: C, 51.12; H, 4.04; N, 10.52. Found: C, 51.34; H, 4.16; N,
10.64.
Compounds 13a and 13c—h were prepared by a procedure similar to that
described for 13b.
3-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thiox-
oimidazolidin-1-yl]propionic Acid (13a) White solid. mp 180—181 °C
(AcOEt–hexane); Rf 0.10 (AcOEt/Hexaneꢀ1 : 1); 1H-NMR (270 MHz,
CDCl3) d: 1.61 (6H, s), 3.01 (2H, t, Jꢀ7.4 Hz), 4.02 (2H, t, Jꢀ7.4 Hz), 7.77
(1H, dd, Jꢀ1.7, 8.3 Hz), 7.88 (1H, d, Jꢀ1.7 Hz), 7.96 (1H, d, Jꢀ8.3 Hz);
MS (ESI) m/z: 386 [(MꢄH)ꢄ]; Anal. Calcd for C16H14F3N3O3S: C, 49.87; H,
3.66; N, 10.90. Found: C, 50.10; H, 3.78; N, 11.13.
5-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thiox-
oimidazolidin-1-yl]pentanoic Acid (13c) White solid. mp 117—118 °C
(AcOEt–hexane); Rf 0.10 (AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz,
CDCl3) d: 1.59 (6H, s), 1.71—1.98 (4H, m), 2.47 (2H, t, Jꢀ7.1 Hz), 3.71
(2H, t, Jꢀ8.1 Hz), 7.76 (1H, dd, Jꢀ1.6, 8.1 Hz), 7.88 (1H, d, Jꢀ1.6 Hz),
7.95 (1H, d, Jꢀ8.1 Hz); MS (ESI) m/z: 414 [(MꢄH)ꢄ]; Anal. Calcd for
C18H18F3N3O3S: C, 52.29; H, 4.39; N, 10.16. Found: C, 52.32; H, 4.45; N,
10.29.
4-{3-[3-(3-Hydroxyphenyl)propyl]-4,4-dimethyl-5-oxo-2-thioxoimida-
zolidin-1-yl}-2-trifluoromethylbenzonitrile (16b) To a solution of com-
pound 15a (91 mg, 0.197 mmol) in CH2Cl2 (2 ml) was added BBr3 (1.0 M in
CH2Cl2, 591 ml, 0.591 mmol) at ꢁ78 °C. The mixture was stirred at ꢁ78 °C
for 10 min and at 0 °C for 40 min. After workup with sat. NaHCO3aq., the
mixture was extracted with CHCl3. Organic layer was washed with brine,
dried over MgSO4, filtered, and concentrated under vacuum. The purification
by silica gel column chromatography (AcOEt/hexaneꢀ1 : 2) gave compound
16b (79.8 mg, 91%) as a colorless oil. Rf 0.50 (AcOEt/hexaneꢀ1 : 1); 1H-
NMR (400 MHz, CDCl3) d: 1.53 (6H, s), 2.13—2.20 (2H, m), 2.69 (2H, t,
Jꢀ7.6 Hz), 3.68—3.70 (2H, m), 4.68 (1H, s), 6.68—6.71 (2H, m), 6.80 (1H,
d, Jꢀ7.3 Hz), 7.18 (1H, t, Jꢀ7.8 Hz), 7.76 (1H, dd, Jꢀ2.0, 8.3 Hz), 7.89
(1H, d, Jꢀ2.0 Hz), 7.95 (1H, d, Jꢀ8.3 Hz); MS (EI) m/z: 447 [Mꢄ].
Compounds 16c and 16d were prepared by a procedure similar to that de-
scribed for 16b.
4-{3-[4-(4-Hydroxyphenyl)butyl]-4,4-dimethyl-5-oxo-2-thioxoimidazo-
lidin-1-yl}-2-trifluoromethylbenzonitrile (16c) Colorless oil. Rf 0.27
(AcOEt/hexaneꢀ1 : 4); 1H-NMR (270 MHz, CDCl3) d: 1.54 (6H, s), 1.60—
1.75 (2H, m), 1.75—1.92 (2H, m), 2.63 (2H, t, Jꢀ7.3 Hz), 3.65—3.71 (2H,
m), 4.71 (1H, s), 6.76 (2H, d, Jꢀ8.4 Hz), 7.06 (2H, d, Jꢀ8.4 Hz), 7.76 (1H,
dd, Jꢀ1.6, 8.3 Hz), 7.88 (1H, d, Jꢀ1.6 Hz), 7.94 (1H, d, Jꢀ8.3 Hz); MS
(ESI) m/z: 462 [MꢄHꢄ].
4-{3-[4-(3-Hydroxyphenyl)butyl]-4,4-dimethyl-5-oxo-2-thioxoimidazo-
lidin-1-yl}-2-trifluoromethylbenzonitrile (16d) Colorless oil. Rf 0.43
(AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.54 (6H, s), 1.69—
1.89 (4H, m), 2.65 (2H, t, Jꢀ7.4 Hz), 3.65—3.71 (2H, m), 4.77 (1H, s),
6.64—6.79 (3H, m), 7.16 (1H, t, Jꢀ7.6 Hz), 7.77 (1H, dd, Jꢀ2.0, 8.3 Hz),
7.89 (1H, d, Jꢀ2.0 Hz), 7.94 (1H, d, Jꢀ8.3 Hz); MS (ESI) m/z: 462
[MꢄHꢄ].
4-{3-[3-(4-Hydroxyphenyl)propyl]-4,4-dimethyl-5-oxo-2-thioxoimida-
zolidin-1-yl}-2-trifluoromethylbenzonitrile (16a) To a solution of com-
pound 14a (440 mg, 0.897 mmol) in 1,4-dioxane (2 ml) was added 6 N-
HCl (4 ml) and the mixture was stirred at 80 °C for 2 h. After cooled to
room temperature, the reaction mixture was extracted with CHCl3. Organic
layer was washed with brine, dried over MgSO4, filtered, and concentrated
under vacuum. The purification by silica gel column chromatography
(AcOEt/hexaneꢀ1 : 3) gave compound 16a (164 mg, 41%) as a colorless oil.
Rf 0.37 (AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.52 (6H, s),
2.07—2.19 (2H, m), 2.66 (2H, t, Jꢀ7.6 Hz), 3.64—3.70 (2H, m), 4.65 (1H,
s), 6.78 (2H, d, Jꢀ8.4 Hz), 7.09 (2H, d, Jꢀ8.4 Hz), 7.76 (1H, dd, Jꢀ2.0,
8.2 Hz), 7.89 (1H, d, Jꢀ2.0 Hz), 7.95 (1H, d, Jꢀ8.2 Hz); MS (EI) m/z: 447
[Mꢄ].
6-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thi-
oxoimidazolidin-1-yl]hexanoic Acid (13d) Colorless oil. 1H-NMR
(270 MHz, CDCl3) d: 1.40—1.55 (2H, m), 1.58 (6H, s), 1.65—1.94 (4H,
m), 2.41 (2H, t, Jꢀ7.1 Hz), 3.66—3.72 (2H, m), 7.77 (1H, dd, Jꢀ1.7,
8.3 Hz), 7.89 (1H, d, Jꢀ1.7 Hz), 7.95 (1H, d, Jꢀ8.3 Hz); MS (ESI) m/z: 428
[(MꢄH)ꢄ]; HR-MS Calcd for C19H21N3O3F3S 428.1250. Found 428.1289.
7-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thi-
oxoimidazolidin-1-yl]heptanoic Acid (13e) Colorless oil. Rf 0.17
(AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.40—1.93 (14H,
m), 2.39 (2H, t, Jꢀ7.1 Hz), 3.62—3.71 (2H, m), 7.76 (1H, d, Jꢀ8.1 Hz),
7.88 (1H, s), 7.95 (1H, d, Jꢀ8.1 Hz); MS (ESI) m/z: 442 [(MꢄH)ꢄ]; HR-
MS Calcd for C20H23N3O3F3S 442.1406. Found 442.1428.
8-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thi-
oxoimidazolidin-1-yl]octanoic Acid (13f) Colorless oil. 1H-NMR
(270 MHz, CDCl3) d: 1.30—1.45 (6H, m), 1.58 (6H, s), 1.58—1.90 (4H,
m), 2.37 (2H, t, Jꢀ7.3 Hz), 3.64—3.70 (2H, m), 7.77 (1H, dd, Jꢀ1.5,
8.2 Hz), 7.89 (1H, d, Jꢀ1.5 Hz), 7.94 (1H, d, Jꢀ8.2 Hz); MS (ESI) m/z: 456
[(MꢄH)ꢄ]; HR-MS Calcd for C21H24F3N3O3S 455.1490. Found 455.1486.
9-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thi-
oxoimidazolidin-1-yl]nonanoic Acid (13g) Colorless oil. Rf 0.27
(AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.30—1.90 (18H,
m), 2.36 (2H, t, Jꢀ7.3 Hz), 3.63—3.69 (2H, m), 7.76 (1H, d, Jꢀ8.2 Hz),
7.88 (1H, s), 7.94 (1H, d, Jꢀ8.2 Hz); MS (ESI) m/z: 470 [(MꢄH)ꢄ]; HR-
MS Calcd for C22H26N3O3F3NaS ([MꢄNa]ꢄ) 492.1539. Found 492.1560.
10-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thi-
oxoimidazolidin-1-yl]decanoic Acid (13h) Colorless oil. 1H-NMR
(270 MHz, CDCl3) d: 1.25—1.42 (10H, m), 1.55—1.73 (2H, m), 1.58 (6H,
s), 1.75—1.80 (2H, m), 2.35 (2H, t, Jꢀ7.4 Hz), 3.63—3.71 (2H, m), 7.76
(1H, dd, Jꢀ1.6, 8.1 Hz), 7.89 (1H, d, Jꢀ1.6 Hz), 7.95 (1H, d, Jꢀ8.1 Hz);
MS (ESI) m/z: 484 [(MꢄH)ꢄ]; HR-MS Calcd for C23H28F3N3O3S 483.1803.
Found 483.1788.
4-{5-Imino-3-[3-(4-methoxymethoxyphenyl)propyl]-4,4-dimethyl-2-
thioxoimidazolidin-1-yl}-2-trifluoromethylbenzonitrile (14a) To a solu-
tion of 5-amino-2-cyanobenzotrifluoride (9) (180 mg, 0.967 mmol) in
THF/H2O (1 : 4, 5 ml) was added thiophosgene (88.5 ml, 1.16 mmol) and the
mixture was stirred at room temperature for 1 h. The mixture was extracted
with AcOEt and the organic layer was washed with brine, dried over MgSO4,
filtered, and concentrated under vacuum to give a crude product of 3-tri-
fluoromethyl-4-cyanoisothiocyanate (10). Acetone cyanohydrin (90%, 149
mg, 1.57 mmol) and 3-(4-methoxymethoxyphenyl)propylamine (280 mg,
1.43 mmol) was dissolved in MeOH (2 ml) and the mixture was stirred at
room temperature for 15 h. Water was added and the mixture was extracted
with AcOEt. The organic layer was washed with brine, dried over MgSO4,
filtered, and concentrated under vacuum to give a crude product of 2-[3-(4-
methoxymethoxyphenyl)propylamino]-2-methylpropionitrile (8i). These two
4-(4-{3-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-
thioxoimidazolidin-1-yl]propyl}phenoxy)butyric Acid Ethyl Ester (17a)
To a solution of compound 16a (57.8 mg, 0.129 mmol) in DMF (1 ml) were

1560
Vol. 56, No. 11
added NaH (60% in oil, 6.2 mg, 0.155 mmol) and ethyl 4-bromobutyrate
(22.4 ml, 0.155 mmol) at 0 °C under N2. After stirring at room temperature
for 30 min, sat.NH4Claq. was added and the mixture was extracted with treated group in which the transcriptional activity is 5% of the transcrip-
value was calculated as the value for firefly luciferase divided by the value
for sea pansy luciferase. The “EC5” value (the concentration of a compound-
AcOEt. Organic layer was washed with brine, dried over MgSO4, filtered
tional activity of 0.1 nM of DHT) was computed to evaluate the agonist activ-
and concentrated under vacuum. The purification by silica gel column chro- ity.
matography (AcOEt/hexaneꢀ1 : 4) gave compound 17a (15.6 mg, 21%) as
Twenty-four hours before transfection, 1.0ꢆ105 HeLa cells were cultured
1
colorless oil. Rf 0.63 (CHCl3); H-NMR (270 MHz, CDCl3) d: 1.26 (3H, t, in phenol red-free DMEM/5% DCC-FBS on 12-well microplates. Five hun-
Jꢀ7.2 Hz), 1.52 (6H, s), 2.08—2.16 (4H, m), 2.51 (2H, t, Jꢀ7.3 Hz), 2.67
dred nanograms/well of MMTV-Luc vector, 100 ng/well of pSG5/hAR, and
(2H, t, Jꢀ7.4 Hz), 3.64—3.70 (2H, m), 3.99 (2H, t, Jꢀ6.1 Hz), 4.15 (2H, q, 5 ng/well of Renilla Luc vector were transfected into the HeLa cells. The
Jꢀ7.2 Hz), 6.83 (2H, d, Jꢀ8.6 Hz), 7.12 (2H, d, Jꢀ8.6 Hz), 7.76 (1H, dd, transfection was performed in a liquid culture of the phenol red-free DMEM
Jꢀ1.7, 8.3 Hz), 7.88 (1H, d, Jꢀ1.7 Hz), 7.94 (1H, d, Jꢀ8.3 Hz) ; MS (EI) using 3 ml/well of lipofectoamine. Nine hours after the transfection, the liq-
m/z: 561 [Mꢄ].
uid culture was replaced by phenol red-free DMEM/3% DCC-FBS contain-
ing 0.1 nmol/l of DHT and 1, 10, 100, 1000, or 10000 nmol/l of test com-
pound. The transcriptional activity was measured 48 h after the replacement
of the liquid culture. Transcriptional activity was measured using a dual lu-
ciferase reporter assay system. The transcriptional activity was calculated by
dividing the value for firefly luciferase by the value for sea pansy luciferase.
4-(4-{3-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-
thioxoimidazolidin-1-yl]propyl}phenoxy)butyric Acid (18a) To a solu-
tion of compound 17a (10.6 mg, 0.0189 mmol) in MeOH (1 ml) was added
2 N-NaOH (0.5 ml) and the mixture was stirred at room temperature 15 h.
The reaction mixture was acidified by 2 N-HCl and extracted with AcOEt.
Organic layer was washed with brine, dried over MgSO4, filtered, and con- The antagonist activity was computed using the following formula and the
centrated under vacuum. The purification by preparative TLC (AcOEt/ antagonist activity determined was used to compute the IC50 value (the con-
hexaneꢀ1 : 1) gave compound 18a (8.4 mg, 83%) as a colorless oil. Rf 0.13 centration of compound that shows a 50% decrease in the transcriptional ac-
(AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.52 (6H, s), 2.02— tivity of DHT 0.1 nmol/l without compound).
2.20 (4H, m), 2.56—2.70 (4H, m), 3.64—3.70 (2H, m), 4.01 (2H, t,
antagonist activity (%)ꢀ(compound-treated transcriptional activity/non-
Jꢀ6.0 Hz), 6.83 (2H, d, Jꢀ8.4 Hz), 7.12 (2H, d, Jꢀ8.4 Hz), 7.76 (1H, dd,
Jꢀ1.8, 8.2 Hz), 7.88 (1H, d, Jꢀ1.8 Hz), 7.94 (1H, d, Jꢀ8.2 Hz); HR-MS
Calcd for C26H26N3O4F3NaS ([MꢄNa]ꢄ) 556.1488. Found 556.1452.
Compounds 18b—d were prepared by a procedure similar to that de-
scribed for 18a.
4-(3-{3-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-
thioxoimidazolidin-1-yl]propyl}phenoxy)butyric acid (18b) Colorless
oil. Rf 0.20 (AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.53
(6H, s), 2.07—2.25 (4H, m), 2.59 (2H, t, Jꢀ7.3 Hz), 2.70 (2H, t, Jꢀ7.3 Hz),
3.66—3.72 (2H, m), 4.02 (2H, t, Jꢀ5.9 Hz), 6.74—6.82 (3H, m), 7.21 (1H,
t, Jꢀ8.6 Hz), 7.76 (1H, dd, Jꢀ1.7, 8.4 Hz), 7.88 (1H, d, Jꢀ1.7 Hz), 7.94
(1H, d, Jꢀ8.4 Hz); MS (EI) m/z: 533 [Mꢄ]; HR-MS Calcd for
C26H26N3O4F3NaS ([M+Na]+) 556.1488. Found 556.1505.
4-(4-{4-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-
thioxoimidazolidin-1-yl]butyl}phenoxy)butyric acid (18c) Colorless oil.
Rf 0.33 (MeOH/CHCl3ꢀ1:20); 1H-NMR (270 MHz, CDCl3) d: 1.54 (6H, s),
1.65—1.73 (2H, m), 1.81—1.85 (2H, m), 2.08—2.15 (2H, m), 2.59 (2H, t,
Jꢀ7.3 Hz), 2.63 (2H, t, Jꢀ7.3 Hz), 3.66—3.70 (2H, m), 4.00 (2H, t,
Jꢀ6.4 Hz), 6.82 (2H, d, Jꢀ8.6Hz), 7.10 (2H, d, Jꢀ8.6Hz), 7.77 (1H, dd,
Jꢀ2.0, 8.3 Hz), 7.89 (1H, d, Jꢀ2.0 Hz), 7.94 (1H, d, Jꢀ8.3 Hz); HR-MS
Calcd for C27H28N3O4F3NaS ([M+Na]+) 570.1644. Found 570.1666.
4-(3-{4-[3-(4-Cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-
thioxoimidazolidin-1-yl]butyl}phenoxy)butyric acid (18d) Colorless oil.
Rf 0.23 (AcOEt/hexaneꢀ1 : 1); 1H-NMR (270 MHz, CDCl3) d: 1.54 (6H, s),
1.65—1.75 (2H, m), 1.81—1.88 (2H, m), 2.09—2.15 (2H, m), 2.59 (2H, t,
Jꢀ6.8 Hz), 2.67 (2H, t, Jꢀ7.6 Hz), 3.67—3.71 (2H, m), 4.02 (2H, t,
Jꢀ6.1 Hz), 6.71—6.74 (2H, m), 6.79 (1H, d, Jꢀ7.8 Hz), 7.20 (1H, t,
Jꢀ8.1 Hz), 7.77 (1H, dd, Jꢀ2.0, 8.3 Hz), 7.89 (1H, d, Jꢀ2.0 Hz), 7.96 (1H,
d, Jꢀ8.3 Hz); HR-MS Calcd for C27H29N3O4F3S ([MꢄH]ꢄ) 548.1825.
Found 548.1852.
treated transcriptional activity)ꢆ100
In Vitro Metabolic Stability in Mouse Liver Microsome To a 0.1 M
potassium phosphate buffer (pH 7.4) containing 1 mM NADPH and
0.5 mg/ml mouse liver microsome, a test compound (final conc. 1 mM) was
added to start the reaction. After incubation for 0, 5, 15, or 30 min at 37 °C,
CH3CN was added to stop the reaction. The concentration of the test com-
pound at each time point was measured by LC/MS/MS (QTRAP, Applied
Biosystems). Hepatic intrinsic clearance (CLint) was determined by the tem-
poral changes in the concentration of the test compound.
In Vivo Antiandrogenic Activities on Seminal Vesicle (SV) Wet
Weights in Mice Immature male ICR mice (7—8 weeks old) were cas-
trated on day 0. After 1 d, animals were divided into groups and dosed on
days 1—4 and 7—10 with one of following: (1) control vehicle (5% gum
arabic, orally); (2) TP (10 mg/body/d, subcutaneously) plus control vehicle;
(3) TP plus compound 13b (2, 10, or 50 mg/kg/d in 5% gum arabic, orally).
Animals were sacrificed on day 11 and SVs were collected and weighed
after formalin fixation. To compare data from different experiments, organ
weights were first standardized as mg/100 g of body weight. Reduction ra-
tios were calculated by the following equation;
reduction ratio (%)ꢀ100ꢆ(BꢁA)/(BꢁC)
A: SV weights of groups under condition (3)
B: SV weights of a group under condition (2)
C: SV weights of a group under condition (1)
Molecular Modeling. Docking Model of Compounds 13b and 13e to
AR This model was built based on the X-ray crystal structure of human
AR in complex with the ligand R1881 (PDB ID: le3g).18) 3D structures of
compounds 13b and 13e were modeled independently using software
SYBYL19) with a Tripos force field.20) Compounds 13b and 13e were manu-
ally docked into AR such that (i) the binding mode of the cyanophenyl moi-
ety of compounds 13b and 13e is similar to that of Casodex (PDB ID:
1z95)21) and (ii) the thiohydantoin ring is modeled so that the side chain at-
tached to the thiohydantoin ring of 13b and 13e is directed to helix 12 of
AR. After checking the bumps between the compound and AR, energy mini-
mization of the compound/AR complex was performed using a molecular
mechanics method with the Tripos force field on condition that the coordi-
nates of AR are fixed. Conformations obtained for 13b and 13e are local
minimum energy conformations.
Competitive Binding Assay CHO-K1/hAR cells (5ꢆ104/well) were
placed in 24-well plates and cultured for 2 d. Adhered cells were washed
with PBS(ꢁ) and replaced with phenol red-free DMEM containing
0.34 nmol/l [3H]-mibolerone in the presence or absence of test compound.
Nonspecific binding of [3H]-mibolerone was determined separately by
adding 200-fold excess of cold mibolerone. Following 2 h incubation at
37 °C, cells were washed with PBS(ꢁ) and solubilized in 10 mmol/l
Tris–HCl (pH 6.8) containing 2% SDS and 10% glycerol. Radioactivity was
counted using a scintillation counter.
The binding affinity was described as an IC50 value (the concentration of a
compound required to inhibit 50% of [3H]-mibolerone).
Reporter Gene Assay Twenty-four hours before transfection, 1.0ꢆ105
HeLa cells were cultured in phenol red-free DMEM/5% DCC-FBS on 12-
well microplates. Five hundred nanograms/well of MMTV-Luc vector,
100 ng/well of pSG5-hAR, and 5 ng/well of Renilla Luc vector were trans-
fected into the HeLa cells. The transfection was performed in a liquid cul-
ture of the phenol red-free DMEM using 3 ml/well of lipofectoamine. Nine
hours after the transfection, the liquid culture was replaced by phenol red-
free DMEM/3% DCC-FBS containing 1, 10, 100, 1000, or 10000 nmol/l of
test compound. The transcriptional activity value was measured 48 h after
the replacement of the liquid culture. Transcriptional activity was measured
with a dual luciferase reporter assay system. The transcriptional activity
Docking Model of Compounds 4 and 18a to AR 3D structures of
compounds 4 and 18a were modeled independently using software SYBYL
with a Tripos force field. Compound 4 was manually docked into AR in a
way that (i) the oxygen atom of the carbonyl group attached at position 3 of
the steroid interacts with the side chain of Arg752 of AR and (ii) the hy-
droxy group attached at position 17 of the steroid interacts with the hydroxy
group of the side chain of Thr877 of AR and (iii) the side chain attached to
position 7a of the steroid core of 4 is directed to helix 12 of AR. The bind-
ing mode of 18a was determined by manual docking in the same way as for
13b and 13e. After checking the bumps between the compound and AR, en-
ergy minimization of the compound/AR complex was performed using a

November 2008
1561
molecular mechanics method with the Tripos force field on condition that
the coordinates of AR are fixed.
(2005).
11) Tachibana K., Imaoka I., Yoshino H., Emura T., Kodama H., Furuta Y.,
Kato N., Nakamura M., Ohta M., Taniguchi K., Ishikura N., Nagamuta
M., Onuma E., Sato H., Bioorg. Med. Chem., 15, 174—185 (2007).
12) Tachibana K., Imaoka I., Yoshino H., Kato N., Nakamura M., Ohta M.,
Kawata H., Taniguchi K., Ishikura N., Nagamuta M., Onuma E., Sato
H., Bioorg. Med. Chem. Lett., 17, 5573—5576 (2007).
Acknowledgments The authors thank Ms. Hitomi Suda of Chugai Phar-
maceutical Co., Ltd. for HRMS measurements of the compounds, and Ms.
Frances Ford of Chugai Pharmaceutical Co., Ltd. for assistance with English
usage.
13) Pike A. C., Brzozowski A. M., Walton J., Hubbard R. E., Thorsell A.
G., Li Y. L., Gustafsson J. Å., Carlquist M., Structure, 9, 145—153
(2001).
14) Teutsch G., Goubet F., Battmann T., Bonfils A., Bouchoux F., Cerede
E., Gofflo D., Gaillard-Kelly M., Philibert D., J. Steroid Biochem. Mol.
Biol., 48, 111—119 (1994).
References
1) Jemal A., Siegel R., Ward E., Murray T., Xu J., Thun M. J., CA Cancer
J. Clin., 57, 43—66 (2007).
2) Stewart A. B., Lwaleed B. A., Douglas D. A., Birch B. R., Curr. Med.
Chem. Anticancer Agents, 5, 603—612 (2005).
3) Kennealey G. T., Furr B. J., Urol. Clin. North Am., 18, 99—110
(1991).
15) Karvonen U., Kallio P. J., Jänne O. A., Palvimo J. J., J. Biol. Chem.,
272, 15973—15979 (1997).
4) Blackledge G., Kolvenbag G., Nash A., Anticancer Drugs, 7, 27—34
(1996).
16) Kemppainen J. A., Langley E., Wong C., Bobseine K., Kelce W. R.,
Wilson E. M., Mol. Endocrinol., 13, 440—454 (1999).
5) Tyrrell C. J., Altwein J. E., Klippel F., Varenhorst E., Lunglmayr G., 17) Kanbe Y., Kim M.-H., Nishimoto M., Ohtake Y., Tsunenari T.,
Boccardo F., Holdaway I. M., Haefliger J. M., Jordaan J. P., J. Urol.,
146, 1321—1326 (1991).
6) Eisenberger M. A., Blumenstein B. A., Crawfold E. D., Miller G.,
Taniguchi K., Ohizumi I., Kaiho S., Nabuchi Y., Kawata S., Morikawa
K., Jo J.-C., Kwon H.-A., Lim H.-S., Kim H.-Y., Bioorg. Med. Chem.
Lett., 16, 4090—4094 (2006).
McLeod D. G., Leohrer P. J., Wilding G., Sears K., Culkin D. J., 18) Matias P. M., Donner P., Coelho R., Thomaz M., Peixoto C., Macedo
Thompson I. M. Jr., Bueschen A. J., Lowe B. A., N. Engl. J. Med., 339,
1036—1042 (1998).
7) Wirth M. P., Froschermaier S. E., Urol. Res., 25 (Suppl 2), S67—S71
(1997).
S., Otto N., Joschko S., Scholz P., Wegg A., Basler S., Schafer M.,
Egner U., Carrondo M. A., J. Biol. Chem., 275, 26164—26171 (2000).
19) “SYBYL 7.3,” Tripos International, St. Louis.
20) Clark M., Cramer III R. D., Van Opdenbosch N., J. Comput. Chem.,
10, 982—1012 (1989).
8) Kemppainen J. A., Wilson E. M., Urology, 48, 157—163 (1996).
9) Ishioka T., Tanatani A., Nagasawa K., Hashimoto Y., Bioorg. Med. 21) Bohl C. E., Gao W., Miller D. D., Bell C. E., Dalton J. T., Proc. Natl.
Chem. Lett., 13, 2655—2658 (2003).
Acad. Sci. U.S.A., 102, 6201—6206 (2005).
10) Hashimoto Y., Miyachi H., Bioorg. Med. Chem., 13, 5080—5093