S. Hosoda et al. / Bioorg. Med. Chem. 14 (2006) 5489–5502
5501
7.15. Measurement of androgen–antagonistic activity by
means of SC-3 growth inhibition assay
(t, J = 7.3 Hz, 6H). 13C NMR (125 MHz, CDCl3/d):
210.1, 153.9, 142.0, 130.5, 125.9, 125.9, 110.1, 69.6,
48.0, 43.2, 40.3, 26.3, 17.3, 16.7, 14.8. HRMS (FAB,
M+) calcd for C33H48O4, 508.3553, found 508.3542.
SC-3 (Shionogi Carcinoma-3) cells were cloned from
Shionogi Carcinoma 115 cells, which were established
from a mouse breast cancer. SC-3 shows androgen-de-
pendent growth.32,33 In this assay, androgenic and
anti-androgenic activities of test compounds were deter-
mined in terms of SC-3 growth promotion and inhibi-
tion, respectively. SC-3 cells were cultured in the
presence of MEM supplemented with 10% FBS and
10 nM testosterone at 37 ꢁC 5% CO2. All experiments
were performed in triplicate or more. For SC-3 cell
growth-inhibition assay, the cells were trypsinized and
diluted to 3.0 · 104 cell/mL with MEM supplemented
with 10% charcoal resin-stripped fetal bovine serum.
This cell suspension was transferred to 96-well microti-
ter plates, and various concentrations of test compound
(from 100 nM to 10 lM DMSO solution) and/or testos-
terone ethanol solution (final concentration 10 nM)
were added. Then the plates were incubated at 37 ꢁC
5% CO2 for 3 days, and the cell number was determined
using the WST-1 method with a Cell Counting Kit and
an MPR-A4i2 micro plate reader (TOSOH, Japan). The
number of cells on wells with testosterone alone was de-
fined as 100%. The concentration of test compounds
that inhibited the increase of the cell number induced
by 10 nM testosterone by 50% was quantified (IC50)
after log–logit transformation. LNCaP growth promo-
tion and inhibition assays were performed by the same
method as described for SC-3 assay, except that
RPMI1640 medium was used.
7.19. 1,1-Bis[4-(3,3-dimethyl-2-oxobutoxy)-3-methylphe-
nyl]cyclobutane (11)
Yield: 87%. White solid. 1H NMR (500 MHz, CDCl3/d):
7.05 (s, 2H), 7.00 (dd, J = 8.6, 2.7 Hz, 2H), 6.52 (d,
J = 8.6 Hz, 2H), 4.81 (s, 4H), 2.64 (t, J = 7.7 Hz, 4H),
2.26 (s, 6H), 1.91 (quint, J = 7.7 Hz, 2H), 1.25 (s,
18H). 13C NMR (125 MHz, CDCl3/d): 210.0, 154.1,
142.8, 128.9, 126.8, 124.1, 110.8, 69.6, 49.9, 43.2, 35.1,
26.3, 16.6, 16.5. HRMS (FAB, M+) calcd for
C30H40O4, 464.2927, found 464.2931.
7.20. 1,1-Bis[4-(3,3-dimethyl-2-oxobutoxy)-3-methylphe-
nyl]cyclopentane (12)
Yield: 67%. White crystals. 1H NMR (500 MHz, CDCl3/
d): 7.01 (s, 2H), 6.98 (dd, J = 8.6, 2.6 Hz, 2H), 6.48 (d,
J = 8.6 Hz, 2H), 4.81 (s, 4H), 2.24 (s, 6H), 2.22–2.18
(m, 4H), 1.67–1.65 (m, 4H), 1.24 (s, 18H). 13C NMR
(125 MHz, CDCl3/d): 210.0, 154.1, 141.8, 129.8, 126.4,
124.8, 110.5, 69.6, 54.3, 43.2, 38.8, 26.4, 23.0, 16.6.
HRMS (FAB, M+) calcd for C31H42O4, 478.3083, found
478.3104.
7.21. 1,1-Bis[4-(3,3-dimethyl-2-oxobutoxy)-3-methylphe-
nyl]cycloheptane (14)
Yield: 88%. White crystals. 1H NMR (500 MHz, CDCl3/
d): 6.92 (s, 2H), 6.87 (dd, J = 8.6, 2.1 Hz, 2H), 6.48 (d,
J = 8.6 Hz, 2H), 4.82 (s, 4H), 2.23 (s, 6H), 2.22–2.20
(m, 4H), 1.66–1.64 (m, 4H), 1.56–1.54 (m, 4H), 1.25 (s,
18H). 13C NMR (125 MHz, CDCl3/d): 210.0, 153.9,
143.9, 130.0, 126.3, 125.1, 110.4, 69.6, 48.7, 43.2, 40.5,
30.1, 26.3, 24.3, 16.7. HRMS (FAB, M+) calcd for
C33H46O4, 506.3396, found 506.3400.
7.16. General procedure for the preparation of 8–14
To a stirred solution of NaH (2.0 equiv/bisphenol) in
DMF (0.3 M) was added bisphenol. The reaction mix-
ture was stirred at 0 ꢁC for 30 min, then 1-chloropinaco-
lone (2.4 equiv) was added and stirring was continued at
room temperature for 3 h. The reaction mixture was
partitioned between aq NH4Cl and ethyl acetate. The
organic layer was washed with brine, dried over MgSO4,
and concentrated. The resulting residue was purified by
silica gel chromatography to afford 8–14.
7.22. 3,3-Bis[4-(3,3-dimethyl-2-oxobutylamino)-3-methyl-
phenyl]pentane (DPP-1111: 15)
1H NMR (500 MHz, CDCl3/d): 6.95 (dd, J = 8.5, 2.1
Hz, 2H), 6.85 (d, J = 2.1 Hz, 2H), 6.39 (d, J = 8.5 Hz,
2H), 4.50 (br, 2H), 4.13 (s, 4H), 2.16 (s, 6H), 1.99 (q,
J = 7.3 Hz, 4H), 1.24 (s, 18H), 0.59 (t, J = 7.3 Hz, 6H).
13C NMR (125 MHz, CDCl3/d): 211.7, 142.6, 138.0,
130.3, 126.5, 121.6, 109.3, 48.9, 48.2, 43.2, 29.5, 26.6,
17.7, 8.6. HRMS (FAB, M+) calcd for C31H46N2O2,
478.3559, found 478.3562.
7.17. 2,2-Bis[4-(3,3-dimethyl-2-oxobutoxy)-3-methylphe-
nyl]propane (8)
Yield: 79%. White crystals. 1H NMR (500 MHz, CDCl3/
d): 6.99 (s, 2H), 6.93 (dd, J = 8.6, 2.6 Hz, 2H), 6.49 (d,
J = 8.6 Hz, 2H), 4.84 (s, 4H), 2.25 (s, 6H), 1.59 (s,
6H), 1.25 (s, 18H). 13C NMR (125 MHz, CDCl3/d):
210.0, 154.1, 143.5, 129.6, 126.4, 124.7, 110.4, 69.5,
43.2, 41.5, 31.0, 26.3, 16.6. HRMS (FAB, M+H+) calcd
for C29H41O4, 453.3005, found 453.3011.
7.23. 1-(4-{3-[4-(3,3-Dimethyl-2-oxobutyl)amino-3-meth-
ylphenyl]pentan-3-yl}-2-methylphenoxy)-3,3-dimethyl-
butan-2-one (DPP-0111: 16)
7.18. 4,4-Bis[4-(3,3-dimethyl-2-oxobutoxy)-3-methylphe-
nyl]heptane (10)
1H NMR (500 MHz, CDCl3/d): 6.93–6.89 (m, 3H), 6.83
(s, 1H), 6.49 (d, J = 8.1 Hz, 1H), 6.39 (d, J = 8.1 Hz,
1H), 4.82 (s, 2H), 4.50 (br, 1H), 4.13 (d, J = 4.3 Hz,
2H), 2.23 (s, 3H), 2.16 (s, 3H), 1.99 (q, J = 7.3 Hz,
4H), 1.25 (s, 9H), 1.24 (s, 9H), 0.59 (t, J = 7.3 Hz,
6H). 13C NMR (125 MHz, CDCl3/d): 211.7, 210.1,
Yield: 60%. White crystals. 1H NMR (500 MHz, CDCl3/
d): 6.91 (s, 2H), 6.87 (dd, J = 8.6, 2.6 Hz, 2H), 6.47 (d,
J = 8.6 Hz, 2H), 4.83 (s, 4H), 2.24 (s, 6H), 1.94–1.90
(m, 4H), 1.25 (s, 18H), 0.96–0.92 (m, 4H), 0.83