New progesterone receptor antagonists: Phosphorus-containing 11β-aryl-substituted steroids

Article abstract of DOI:10.1016/j.bmc.2006.05.066

A new series of phosphorus-containing 11β-aryl-substituted steroids have been synthesized in an eight-step sequence involving a palladium-catalyzed coupling reaction to introduce a phosphorus group onto the aromatic ring. The compounds were evaluated for

Full text of DOI:10.1016/j.bmc.2006.05.066

Bioorganic & Medicinal Chemistry 14 (2006) 6726–6732
New progesterone receptor antagonists: Phosphorus-containing
11b-aryl-substituted steroids
Weiqin Jiang,* George Allan, James J. Fiordeliso, Olivia Linton, Pamela Tannenbaum,
Jun Xu, Peifang Zhu, Joseph Gunnet, Keith Demarest, Scott Lundeen and Zhihua Sui
Johnson & Johnson Pharmaceutical Research & Development L.L.C., Drug Discovery, 1000 Route 202, Raritan, NJ 08869, USA
Received 16 March 2006; revised 22 May 2006; accepted 30 May 2006
Available online 27 June 2006
Abstract—A new series of phosphorus-containing 11b-aryl-substituted steroids have been synthesized in an eight-step sequence
involving a palladium-catalyzed coupling reaction to introduce a phosphorus group onto the aromatic ring. The compounds were
evaluated for progesterone receptor (PR) antagonist activity in a T47D cell-based assay and for glucocorticoid receptor (GR) antag-
onist activity in an A549 cell-based assay. The structure–activity relationships of these compounds are discussed. Selected com-
pounds were tested in vivo in a rat complement C3 assay.
ꢀ 2006 Elsevier Ltd. All rights reserved.
1. Introduction
Steroid receptors are closely related structurally and in
their mechanism of action. Slight modifications of the
steroidal structure have been found to induce important
affinity and specificity variations for the corresponding
receptors. The most prominent structural feature of
mifepristone is the 4-(dimethylamino)phenyl group at
the 11b-position of the 19-nor steroid. Without this sub-
stituent, the molecule would be expected to act as a pro-
gestin. Replacement of the 4-(dimethylamino)phenyl
group with a 4-acetylphenyl (ZK112993, 2)3 leads to
equally potent or more potent antiprogestins. Both com-
pounds 1 and 2 are known for their overt antiglucocor-
ticoid activity, which prevents their long-term clinical
usage. Minor changes at the C-17 position produce anti-
progestins with dramatically reduced antiglucocorticoid
activity.4 For example, Org-33628 (compound 3) with a
spiral cyclic chain at the C-17 position exhibits higher
PR antagonist potency with significantly lower GR
activity,2a,2b when compared with mifepristone (1) and
ZK112993 (2). This led us to use Org-33628 as a tem-
plate to make a chemically novel steroidal series with
similar PR potency and higher selectivity against GR,
when compared to mifepristone (compound 1).
The progesterone receptor (PR) is a member of the ste-
roid receptor sub-family of the nuclear hormone recep-
tor superfamily, a group of nuclear transcription
factors.1 Progesterone, the endogenous ligand for the
PR, is involved in the control of ovulation and prepara-
tion of the uterus to support pregnancy. A PR antago-
nist therefore has potential utility as a contraceptive.
In addition, PR antagonists have potential applications
in the treatment of reproductive disorders such as uter-
ine leiomyomas and endometriosis, as well as hormone
dependent tumors.1
The discovery of the first competitive progesterone
antagonist, mifepristone (Fig. 1, RU-486, 1), has stimu-
lated an intense search for more potent and more selec-
tive antiprogestins.2 Although various attempts to
differentiate the antiprogestin activity of mifepristone
and other analogues from their antiglucocorticoid activ-
ity have identified some selective progesterone receptor
modulators (PRMs), new compounds having antipro-
gestational activity devoid of antiglucocorticoid activity
remain highly desirable, both in terms of clinical appli-
cations and basic endocrine research.2a,2c
There have been literature reports on using phosphinic
acids as bioisosteres of the carboxylic acid group.5 We
envisioned that phosphonyl groups can serve as bioisos-
teres for the carbonyl group on the 11-b-aryl group of
Org-33628. In the present article, we describe the synthe-
sis and biological properties of these derivatives.
Keywords: Phosphorus-containing steroids; Progesterone receptor
antagonist.
*
Corresponding author. Tel.: +1 609 409 3496; fax: +1 609 655
6930; e-mail: wjiang1@prdus.jnj.com
0968-0896/$ - see front matter ꢀ 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2006.05.066

W. Jiang et al. / Bioorg. Med. Chem. 14 (2006) 6726–6732
6727
O
O
N
O
HO
HO
O
H
H
H
H
H
H
3
O
O
1
2
Mifepristone
(RU-486)
Org-33628
ZK112993
Figure 1. Some known steroidal progesterone receptor antagonists.
2. Chemistry
THPO
O
The backbone of the steroid was assembled according to
the route described in Scheme 1.6a Thus, lithiated meth-
oxy allenamide was captured by commercially available
ethylene deltanone 11, to generate compound 12. Base-as-
sisted cyclization was accomplished using a catalytic
amount of crown ether (di-cyclohexyl-18-crown-6) to
generate compound 13. The selected deprotection of the
methoxy group was accomplished by carefully monitor-
ing the reaction time and the amount of acid used for
the reaction. The resulting compound 14 was subjected
to Wittig reaction condition to furnish compound 15.
At this point, the C-17 side chain for Org-33628 is fully
installed.
BrMg
p-TSA
78%
OTHP
O
16
CuCl, THF
53%
O
17
OH
HO
TfO
O
O
Tf2NPh
NaH
64%
O
O
19
18
Epoxidation using mCPBA under basic conditions pro-
vided epoxide intermediate 16, with a:b ratio of 3:1,
which is in consistency with the literature.6b The remain-
ing products resulted from epoxidation of either one or
both of the double bonds between C9 to C11 and the
double bond on the C-17 spiral side chain. These side
products are separable from the desirable product. In
Scheme 2, epoxide 16 was subjected to the cuprate reac-
tion conditions to provide the intermediate 17.6b Free
phenol 18 was obtained upon acidic deprotection condi-
tions and then was reacted with a triflating reagent
(Tf2NPh) to furnish aryl triflate 19.7 For the conversion
of compound 17 to 18, both strong acid p-TSA and mild
acid, such as, oxalic acid or acetic acid work the same.
Scheme 2. Installation of C11 side chain and preparation of triflate
precursor 19.
In the literature, aryl bromides, iodides and triflates can
react with dialkyl phosphites or dialkyl phosphine oxide
catalyzed by palladium/phosphine ligand.8a,b,c Since an
iodide or bromide group cannot remain intact in the
Grignard precursor for the cuprate reaction, we chose
to use triflate 19 as the precursor for the phosphination
reaction. There are several examples in the literature on
converting the triflate group to a phosphonyl group. We
first examined Pd(PPh3)4/4-methyl-morpholine using
conventional synthesis (100 ꢁC, 16 h) as well as with
O
OH
C
O
O
O
ii
i
O
O
O
O
O
12
O
13
11
O
O
O
O
iii
iv
v
O
O
O
O
16
15
O
O
O
14
Scheme 1. Installation of C-17 side chain. Reagents: (i) n-BuLi, 1-methoxy-propa-1,2-diene 5, ꢀ78 ꢁC; (ii) KO-t-Bu, Di-Cy-18-C-6 7, t-BuOH; (iii)
HCl/acetone; 43% for last 3 steps; (iv) CH3(PPh3)Br, KO-t-Bu, toluene, 67%; (v) mCPBA, sodium bicarbonate, CH2Cl2, 38%.

6728
W. Jiang et al. / Bioorg. Med. Chem. 14 (2006) 6726–6732
microwave irradiation (30 min, 150 ꢁC). The product
was formed with low conversion. The second catalyst
system, Pd(OAc)2/dppp/DIEA, turned out to be practi-
cal in generating clean product with a facile purification.
Due to the switch from a monodentate ligand PPh3 to a
bidentate ligand 1,4-bis(diphenylphosphino)butane
(dppp), the reductive elimination step in the catalytical
cycle is probably accelerated due to the faster dissocia-
tion of the bidentate phosphine ligand, dppp, than that
of the monodentate ligand PPh3, in a dissociative mech-
anism in catalytical cycle.9 Despite the abundant litera-
tures on the thermal reaction of phosphination from
aryl or vinyl triflate,8 the reaction involving irradiation
by microwaves has not been reported before. We found
that microwave irradiated conditions constantly provid-
ed the desired coupling product 20 with higher yield and
purity. It is observed that THF, 1,4-dioxane, and DMF
are all suitable solvents, while DMSO tends to be unsta-
ble under microwave conditions Scheme 3.
from a glucocorticoid response element (GRE)-linked
luciferase reporter gene in the human lung carcinoma
cell line A549. The IC50s of the compounds from the
T47D and A549 assays are listed in Table 1. The ratio
of the A549 IC50 to the T47D IC50 was calculated and
is listed in the column ‘Ratio’, as a measure of the
separation of PR and GR antagonist activities.
Mifepristone (RU-486), a commercial drug, was tested
as a control.
The progesterone antagonist activities in the T47D assay
were first examined. A wide range of substitutions at R1
and R2 are tolerated, except for acidic functional groups
such as OH. When R1 and R2 are alkoxy groups, the size
change did not affect potencies, as shown in 20a and 20c.
Yet, a change in the electronics reduced potency slightly,
as shown in 20a and 20 g, where 20g bears a more elec-
tron-withdrawing group. Substitution of the phenyl
group did not significantly change the potency (20 h vs
20i). Replacing the methyl with phenyl group decreased
the potency (20b vs 20h). When R1 and R2 are tied into a
cyclic structure, such as in 20k, the potency remained
unchanged in comparison with 20a. Finally, replacing
MeO with OH (20c vs 20d) or EtO with OH (20e vs
20f) abolished T47D activity. The poor antagonistic
activities of 20d and 20f might be caused by the negative
charge of these compounds which prevents them from
penetrating the membrane and getting into the cell.
Although our compounds were less potent than mifepri-
stone, representative compounds demonstrated better
selectivity (ꢁ13-fold), such as 20b and 20e. In terms of
potency, more modification of this series of compounds
needs to be done in order for them to be of value for fur-
ther drug development.
By using LiOH/tBuOH/water,10a compounds 20c and
20e were converted to mono-hydroxyl phosphoryl 11-
b-phenyl-substituted steroids 20d and 20f. We also
10b
attempted the same conversion by using KOSiMe3,
only to recover starting material. The most commonly
used acidic reagent TMSBr10c led to an A-ring aroma-
tized compound without any formation of desired prod-
uct (Scheme 4).
3. Results and discussion
The compounds were evaluated for PR antagonist activ-
ity based on their ability to block progesterone induc-
tion of alkaline phosphatase activity in the human
breast cancer cell line T47D (Table 1). The compounds
were also tested for their GR antagonist activity based
on their ability to inhibit corticoid-induced transcription
Compounds 20a, 20b, and 20e were further evaluated
in vivo in the rat uterus complement C3 assay.11 In this
assay, ethinyl estradiol (EE) stimulates C3 expression
and progestins inhibit this expression. In turn, antipro-
gestins counteract inhibition by the progestin. The
results was shown in Table 2. The in vivo activities of
three selected compounds 20a, 20b, and 20e showed
similar trends as in the T47D assay.
O
R1
P
O
R2
Pd(OAc)2, dppp
DIEA, 1,4-dioxane
19
H(O)PR1R2
4. Conclusion
19 ~ 52 %
microwave
O
20
In summary, a novel series of phosphorus-containing
C11 aryl-substituted steroids were synthesized by
utilizing Pd-catalyzed phosphination reaction of triflate.
Scheme 3. Pd-catalyzed phosphination reaction under microwave
irradiation.
O
O
R1
P
R1
P
O
O
R2
HO
LiOH
tBuOH, H2O
O
O
20d R1 = OMe
20f R1 = Me
20c R1=R2=OMe
20e R1=Me, R2 = OEt
19%
34%
Scheme 4. Synthesis of 20d and 20f via hydrolysis reaction.

W. Jiang et al. / Bioorg. Med. Chem. 14 (2006) 6726–6732
Table 1. Potencies of compounds 20a–20k in T47D and A549 assays
6729
O
P
R1
O
R2
O
Compound
R1
R2
—
T47D IC50 (nM)
A549 IC50 (nM)
Ratio = IC50
(A549)/IC50 (T47D)
1
Mifepristone
OCH2CH3
0.2
9.9
2.6
237.7
13
24
184
28
—
169
—
5
20a
20b
20c
20d
20e
20f
20g
20h
20i
OCH2CH3
Me
Me
OCH3
3.58
8.27
>1000
1.6
660.15
230.1
OCH3
OH
OCH3
Me
>3000
270.86
>3000
140.81
163.45
143.11
34.33
OCH2CH3
OH
Me
OCH2CF3
>1000
28
44
OCH2CF3
Ph
Ph
4-Cl-Ph
4
4-Cl-Ph
PhO
—
20
7
20j
20k
PhO
–OCH2CMe2CH2O–
3.4
7.2
10
40
286.53
Table 2. Potencies of compounds 20a, 20b, and 20e tested in the rat
uterine C3 model by oral administration
buffer, assay buffer, and 1:20 substrate/enhancer mixture
were then added. After a 1-h incubation at room
temperature in the dark, the lysate was transferred to
a white 96-well plate (Dynex) and luminescence was
read using a LuminoSkan Ascent (Thermo Electron,
Woburn, MA).
Compound
% inh
ID50 (mg/kg)
Mifepristone11
3.0
>30
>30
20a
20b
20e
16% inh at 30 mpk
38% inh at 20 mpk
18% inh at 10 mpk
78% inh at 30 mpk
10–30
5.1.2. A549 reporter assay. A549 human lung carcinoma
cells were washed with OPTI-MEM I (Gibco). The
medium was removed and lipid–DNA complex solution
(1.5 lg/mL of GRE-luciferase reporter DNA in 8.5 mL
OPTI-MEM I plus 6 lL/mL DMRIE-C reagent in
8.5 mL OPTI-MEM I, combined and mixed, then incu-
bated at room temperature for 40 min) was overlayed
onto the cells in a T160 flask. The cells were incubated
for 16 h at 37 ꢁC in a CO2 incubator. The DNA-contain-
ing medium was removed and 30 mL of growth medium
containing 5% (v/v) charcoal-treated fetal bovine serum
was added. After 5–6 h, the cells were seeded in 96-well
plates and incubated overnight at 37 ꢁC. To each well
were then added test compounds followed by dexameth-
asone as a corticoid challenge. The cells were incubated
for 24 h. Luciferase assay buffer (Promega) was added to
each well and the cells were incubated for 30 min at
room temperature. Luciferase activity was measured in
a DYNEX Microlite plate on a TopCount (Packard).
These compounds were tested in cell-based in vitro
assays for progestin and glucocorticoid antagonist activ-
ities. Most of the compounds were potent PR antago-
nists (nanomolar range), with some showing better
selectivity than mifepristone. Selected compounds
showed modest oral progestin antagonist activity in rat
uterus.
5. Experimental
5.1. Biological assay
5.1.1. T47D alkaline phosphatase assay. T47D human
breast cancer cells were plated in 96-well tissue culture
plates at 10,000 cells per well in assay medium [RPMI
medium without phenol red (Invitrogen) containing
5% (v/v) charcoal-treated FBS (Hyclone) and 1% (v/v)
penicillin–streptomycin (Invitrogen)]. Two days later,
the medium was decanted and test compound or control
was added at a final concentration of 0.1% (v/v) dimeth-
ylsulfoxide in fresh assay medium. Twenty-four hours
later, an alkaline phosphatase assay was performed
using a SEAP kit (BD Biosciences Clontech, Palo Alto,
CA). Briefly, the medium was decanted and the cells
were fixed for 30 min at room temperature with 5%
(v/v) formalin (Sigma). The cells were washed once at
room temperature with Hanks’ buffered saline solution
(Invitrogen). Equal volumes (0.05 mL) of 1· dilution
5.1.3. Rat complement C3 assay. Ovariectomized two-
month-old Sprague–Dawley rats were purchased from
Harlan (Indianapolis, IN). Five to seven days after sur-
gery, the rats were dosed once with test compound or
control. All of the animals received ethinyl estradiol at
a dose of 0.3 mg/kg, administered orally as a suspension
in sesame oil. In addition, the animals received a dose of
3 mg/kg medroxyprogesterone acetate (progestin ago-
nist), given orally in 20% (w/v) hydroxyl-propyl-b-D-cy-
clodextrin. Test compounds were dosed orally in
cyclodextrin, while this vehicle was used as the negative

6730
W. Jiang et al. / Bioorg. Med. Chem. 14 (2006) 6726–6732
control (no progestin antagonism). Mifepristone was
given as a positive control for antagonism. About 24 h
later, the rats were euthanized by carbon dioxide
asphyxiation. Whole uteri were removed, trimmed of
fat, and frozen on dry ice prior to storage at ꢀ80 ꢁC.
The uteri were homogenized in 1–2 mL each of TRIzol
(Invitrogen Life Technologies, Carlsbad, CA); and the
homogenates were processed for RNA preparation
according to the manufacturer’s directions. Quantitative
PCR was performed using rat complement C3 primers
and TaqMan probe from Applied Biosystems (Foster
City, CA) and an ABI PRISM 7000 Sequence Detection
System (Applied Biosystems). The level of 28S ribosom-
al RNA in each sample was determined for normaliza-
tion, and a dilution series of one of the estrogen-
treated samples was used to generate a standard curve.
In Table 2, percent inhibition and ID50 data are shown.
This is a measure of the extent of inhibition of the pro-
gestin effect by the antagonist. To calculate these values,
uterus weight in the presence of estradiol plus the ago-
nist was set at 0% inhibition (baseline for an antagonist),
and uterus weight in the presence of estradiol alone was
set at 100% inhibition.
round-bottomed flask. To this solution was added sodi-
um bicarbonate (5.38 g, 64.0 mmol) and the mixture was
cooled to less than ꢀ40 ꢁC under nitrogen using a dry
ice-acetone bath. A solution of mCPBA (75%, 2.87 g,
12.48 mmol) in dichloromethane (40 mL) was prepared
and added to the reaction mixture in portions via syr-
inge. The reaction mixture was stirred at less than
ꢀ40 ꢁC for half an hour, then stirred in an ice bath for
half an hour. The reaction mixture was partitioned in
a mixture of ice water/dichloromethane (1:1). After stir-
ring, the layers were separated upon melting of the ice
and the aqueous layer was extracted twice with dichloro-
methane. The organic layers were washed with saturated
sodium bicarbonate, water, brine, dried, filtered, and
evaporated to provide a clear oil that later solidified.
The material was purified by column chromatography
(5–30% ethyl acetate/hexane) to provide desired product
as a white solid (1.6 g, 38%). 1H NMR (400 MHz,
CDCl3) d 5.96 (br s, 1H), 5.03 (s, 1H), 4.79 (s, 1H),
3.96–3.75 (m, 6H), 2.64–2.60 (m, 2H), 2.43 (m, 1H),
2.17–1.12 (m, 17H), 0.87 (s, 3H); MS (m/e): 385 (MH+).
5.2.4. 19,24-Dinorchola-9,20-dien-3-one, 11-[4-(2-tetrahy-
dro-2-H-pyranoxy)phenyl]-17,23-epoxy-5-hydroxy-, cyclic
1,2-ethanediyl acetal, (5a,11b,17a)-(9CI) (17). Copper (I)
chloride (257 mg, 2.60 mmol) was weighed into a
100-mL round-bottomed flask. After flushed with dry
nitrogen and capped with septum, 4-(2-tetrahydro-2-
H-pyranoxy)-phenylmagnesium bromide (0.5 M in
THF) (10.4 mL, 5.20 mmol) was added via syringe un-
der nitrogen. Solution was stirred vigorously for
1–3 min until a cloudy, white solution resulted. Immedi-
ately, a solution of compound 16 (1.0 g, 2.60 mmol) in
THF (15 mL) was added. The mixture was stirred for
1 h and then was quenched by adding aqueous saturated
ammonium chloride solution and was extracted twice
with ethyl acetate. The organic extracts were washed
with water, brine, dried, filtered, and evaporated to a
residue. The residue was purified by column chromatog-
raphy (5–40% ethyl acetate/hexane) to afford desired
product as a white solid (0.77 g, 53%). 1H NMR
(400 MHz, CDCl3) d 7.07 (d, J = 8.6 Hz, 2H), 6.92 (d,
J = 8.7 Hz, 2H), 5.34 (m, 1H), 5.08 (s, 1H), 4.82
(s, 1H), 4.35 (s, 1H), 4.15 (s, 1H), 4.0–3.89 (m, 5H),
3.81–3.76 (m, 2H), 3.60 (m, 1H), 2.63 (m, 2H), 2.40–
1.24 (m, 24H), 0.53 (s, 3H); MS (m/e): 585 (MNa+),
545(MꢀH2O)+.
5.2. General information for chemistry
1H NMR spectra were obtained at 400 MHz and
300 MHz on Brucker AVANCE300 and AVANCE400
spectrometer. Chemical shifts are reported in ppm down-
field from TMS as an internal standard. Thin-layer chro-
matography was carried out using 2.5 · 7.5-cm silica gel
60 (250 lM layer) plates with UV detection. Magnesium
sulfate was employed to dry organic extracts prior to
concentration by rotary evaporation. Flash chromatog-
raphy was done using EM science silical gel 60 (230–
400 mesh). Standard solvents from J. T. Baker were used
as received. Anhydrous solvents from J. T. Baker or
Aldrich and all other commercially available reagents
were used without further purification. Mass spectra
were obtained on a Hewlett-Packard 5989A quadrupole
mass spectrometer. Silica gel (E. Merck, 230–400 mesh)
was used for all flash chromatography. Thin-layer chro-
matography was performed on Analtech silica gel GF
prescored plates (250 lm). HPLC analysis was carried
out on Agilent 1100 Series LC/MSD equipment.
5.2.1.
Spiro[estra-5(10),9(11)-diene-17,20(30H)-furan]-
3,30-dione,40,50-dihydro-, cyclic 3-(1,2-ethanediyl acetal),
(17b)-(9CI) (14). Compound 14 was prepared from eth-
ylene deltanone (11) in 3 steps according to the proce-
dure reported in Gange, D.; Magnus, P. J. Am. Chem.
Soc. 1978, 7746–7747.
5.2.5. 19, 24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-(4-hydroxyphenyl)-, (11b,17a)-(9CI) (18). A solution
of compound 17 (100 mg, 0.178 mmol) in acetone
(10 mL) was prepared. Oxalic acid (22 mg, 0.178 mmol)
was added. The reaction mixture was heated to reflux.
After 1 h, an additional amount of oxalic acid (17 mg)
was added and refluxed for one more hour. Water was
added and the solution was extracted twice with ethyl
acetate. The organic extracts were dried, filtered, and
evaporated to yield a white solid. The crude material
was purified by column chromatography (5–50% ethyl
acetate/hexane) to afford a white solid (58 mg, 78%).
1H NMR (400 MHz, CDCl3) d 6.99 (d, J = 8.5 Hz,
2H), 6.73 (dd, J = 1.9 and 6.7 Hz, 2H), 5.77 (s, 1H),
5.44 (s, 1H), 5.13 (s, 1H), 4.84 (s, 1H), 4.23 (d, J = 7.1
5.2.2. 19,24-Dinorchola-5(10),9(11),20-trien-3-one, 17,23-
epoxy-, cyclic 1,2-ethanediyl acetal, (17a)-(9CI) (15). This
compound was prepared according to the procedure
reported in Hamersma, J. A.; Orlemans, E. O. M.;
Rewinkel, J. B. M. EP0582338A2.
5.2.3. 19,24-Dinorchola-9(11),20-dien-3-one, 5,10:17,23-
diepoxy-, cyclic 1,2-ethanediyl acetal, (5a,10a,17a)-(9CI)
(16). A solution of compound 15 (4.0 g, 10.85 mmol) in
dichloromethane (40 mL) was prepared in a 500-mL

W. Jiang et al. / Bioorg. Med. Chem. 14 (2006) 6726–6732
6731
Hz, 1H), 3.87–3.80 (m, 2H), 2.69–1.24 (m, 18H), 0.59 (s,
(m, 2H), 5.71 (s, 1H), 5.12 (s, 1H), 4.81 (s, 1H), 4.31 (d,
1H, J = 5.9 Hz), 3.75 (m, 8H), 3.5 (s, 1H), 2.68–1.38 (m,
17H), 0.52 (s, 3H); MS (m/e): 509(MH+), 531 (MNa+),
1017 (2MH+).
3H); MS (m/e): 439 (MNa+), 417 (MH+).
5.2.6. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-(4-trifluoromethanesulfonyloxyphenyl)-, (11b,17a)-
(9CI) (19). To a dry flask containing NaH (35 mg,
60% in mineral oil, 0.864 mmol) was added a solution
of compound 18 (0.24 g, 0.576 mmol) in THF (20 mL)
slowly. The solution was cooled in a dry-ice acetone
bath under nitrogen. After 30 min, N-phenyl-trifluorom-
ethane-sulfonimide (309 mg, 0.864 mmol) in THF
(3 mL) was added. After 35 min, the mixture was slowly
warmed to room temperature. This was stirred until
reaction was completed, monitored by TLC and HPLC.
Brine was added followed by ethyl acetate. The aqueous
layer was extracted with ethyl acetate and the organic
layers were washed with brine, dried, filtered, and evap-
orated. The crude material was purified by column chro-
matography (5–30% ethyl acetate/hexane) to afford a
white solid (250 mg, 64%). 1H NMR (400 MHz, CDCl3)
d 7.26–7.18 (m, 4H), 5.79 (s, 1H), 5.15 (t, J = 1.8 Hz,
1H), 4.86 (s, 1H), 4.32 (d, J = 7.1, 1H), 3.87–3.77 (m,
2H), 2.72–2.56 (m, 5H), 2.48–1.24 (m, 13H), 0.54 (s,
3H); MS (m/e): 549 (MH+), 571 (MNa+).
5.2.10. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-(4-(hydroxy-methoxy-phosphorylphenyl)-, (11b,17a)-
(9CI) (20d). Compound 20c (80 mg, 0.157 mmol) in
t-BuOH (2.4 mL) and water (1.2 mL) with LiOH
(8 mg, 0.314 mmol) was stirred at 80 ꢁC for 2.5 h. The
resulted solution was partitioned between ethyl ace-
tate/water (50 mL:50 mL). The organic layer was dried
and concentrated. The resulted crude material was puri-
fied by preparative TLC (30% methanol/dichlorometh-
ane) to provide desired product (14 mg, 19%). 1H
NMR d (CD3OD) 7.70 (m, 2H), 7.21 (m, 2H), 5.70 (s,
1H), 5.18 (s, 1H), 4.42 (d, 1H, J = 5.8 Hz), 3.80 (m,
3H), 3.42 (m, 4H), 2.78–1.08 (m, 17H), 0.58 (s, 3H);
MS (m/e): 495(MH+), 517 (MNa+), 1011 (2MNa+).
5.2.11. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-[4-(ethoxy-methyl-phosphinoyl)phenyl]-, (11b,17a)-(9CI)
(20e). The title compound was prepared as a white solid
according to the procedure for compound 20b in 52%
yield. 1H NMR (400 MHz, CDCl3) d 7.72 (m, 2H),
7.29 (m, 2H), 5.79 (s, 1H), 5.18 (s, 1H), 4.86 (s, 1H),
4.36 (d, 1H, J = 5.7 Hz), 4.08 (m, 2H), 3.82 (m, 2H),
3.4 (s, 1H), 2.71–1.22 (m, 23H), 0.52 (s, 3H); MS (m/
e): 507 (MH+), 529 (MNa+); HRMS: calcd MH+ for
C31H39O4P 507.2664; found 507.2666.
5.2.7. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-(4-(diethoxy-phosphorylphenyl)-, (11b,17a)-(9CI) (20a).
The title compound was prepared as a white solid (20 mg,
41%) according to the procedure for compound 20b, start-
1
ing from compound 19 (50 mg, 0.091 mmol). H NMR
(400 MHz, CDCl3) d 7.72 (dd, J = 8.2 and 12.0 Hz, 2H),
7.28–7.26 (m, 2H), 5.78 (s, 1H), 5.14 (s, 1H), 4.86
(s, 1H), 4.34 (d, J = 7.3 Hz, 1H), 4.17–4.05 (m, 4H),
3.85–3.79 (m, 2H), 2.70–2.58 (m, 5H), 2.49–1.24 (m,
19H), 0.53 (s, 3H); MS (m/e): 536 (MH+), 559 (MNa+).
5.2.12. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-[4-(hydroxy-methyl-phosphinoyl)phenyl]-, (11b,17a)-
(9CI) (20f). The title compound was prepared as a white
solid according to the procedure for compound 20d in
1
34% yield. H NMR (CD3OD) 7.68 (m, 2H), 7.21 (m,
5.2.8. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-(4-(dimethyl-phosphorylphenyl)-, (11b,17a)-(9CI)
(20b). A mixture was prepared consisting of compound
19 (150 mg, 0.273 mmol), dimethylphosphine oxide
(43 mg, 0.547 mmol), palladium (II) acetate (6.1 mg,
0.0273 mmol), 1,4-bis(diphenylphosphino)butane (17 mg,
0.041 mmol), diisopropylethylamine (0.19 mL, 1.092
mmol), and dioxane (3.5 mL). The mixture was reacted
using a CEM microwave utilizing the P150 program at
150 ꢁC for 30 min. The reaction was completed by LC-
MS. The orange solution was poured onto water and
extracted twice with ethyl acetate. The organic extracts
were washed with brine, dried, filtered, and evaporated.
The residue was purified by column chromatography (5–
40% methanol/ethyl acetate) to afford a white solid
(65.6 mg, 50%). 1H NMR (400 MHz, CDCl3) d 7.67–
7.61 (dd, J = 8.5 and 11.4 Hz, 2H), 7.31–7.26 (dd,
J = 2.0 and 8.3 Hz, 2H), 5.78 (s, 1H), 5.15 (t, J = 1.8 Hz,
1H), 4.86 (s, 1H), 4.34 (d, J = 7.0 Hz, 1H), 3.85–3.79 (m,
2H), 3.49 (m, 1H), 2.73–2.53 (m, 5H), 2.49–0.80 (m,
18H), 0.55 (s, 3H); MS (m/e): 477 (MH+), 500 (MNa+).
2H), 5.72 (s, 1H), 5.68 (s, 1H), 4.42 (d, 1H), 3.80 (m,
2H), 2.78–1.29 (m, 22H), 0.56 (s, 3H); MS (m/e): 477
(MHꢀ), 501 (MNa+); HRMS: calcd MH+ for
C29H35O4P 479.2351; found 479.2361.
5.2.13. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-ep-
oxy-11-[4-[bis-(2,2,2-trifluoro-ethoxy)-phosphoryl]-phen-
yl]-, (11b,17a)-(9CI) (20 g). The title compound was
prepared as a white solid (107 mg, 30%) according to
the procedure for compound 20b, starting from com-
1
pound 19 (300 mg, 0.547 mmol). H NMR (CDCl3) d
7.72 (m, 2H), 7.32 (m, 2H), 5.75 (s, 1H), 5.13 (s, 1H),
4.83 (s, 1H), 4.45 (m, 5H), 3.81 (m, 2H), 2.71–1.34 (m,
18H), 0.52 (s, 3H); MS (m/e): 645 (MH+), 667
(MNa+); HRMS: calcd MH+ for C32H35F6O5P
645.2205; found 645.2171.
5.2.14. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-11-
[4-(diphenyl-phosphinoyl)-phenyl]-, (11b,17a)-(9CI) (20h).
The title compound was prepared as a white solid product
(104 mg, 47%), according to the procedure for compound
20b, starting from compound 19 (200 mg, 0.365 mmol).
1H NMR (CDCl3) d 8.02 (m, 2H), 7.62–7.42 (m, 12H),
5.72 (s, 1H), 5.12 (s, 1H), 4.82 (s, 1H), 4.33 (d, 1H,
J = 5.9 Hz), 3.81 (m, 2H), 2.71–1.42 (m, 18H), 0.52 (s,
3H); MS (m/e): 601 (MH+), 623 (MNa+); HRMS: calcd
MH+ for C40H41O3P 601.2872; found 601.2855.
5.2.9. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-epoxy-
11-(4-(dimethoxy-phosphorylphenyl)-,
(11b,17a)-(9CI)
(20c). The title compound was prepared as a white solid
according to the procedure for compound 20b in 63.4%
yield. 1H NMR (400 MHz, CDCl3) d 7.71 (m, 2H), 7.21

6732
W. Jiang et al. / Bioorg. Med. Chem. 14 (2006) 6726–6732
5.2.15. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-ep-
oxy-11-[4-[bis-(4-chloro-phenyl)-phosphinoyl]-phenyl]-,
(11b,17a)-(9CI) (20i). The title compound was prepared
as a white solid (155 mg, 42%) according to the proce-
dure for compound 20b, starting from compound 19
References and notes
1. Teutsch, G.; Ojasoo, T.; Raynaud, J. P. J. Steroid
Biochem. 1988, 31, 549–565, and references therein.
2. (a) Teutsch, G.; Philibert, D. Hum. Reprod. 1994(Suppl.
1), 11–30; (b) Orlemans, E. O. M. Oral Communication at
the 206th Annual Meeting of the American Chemical
Society, Chicago, 1993, August 22–27; (c) Michna, H.;
Parczyk, K.; Schneider, M. R.; Nishino, Y. Ann. N.Y.
Acad. Sci. 1995, 761, 224–247.
1
(300 mg, 0.547 mmol). H NMR d (CDCl3) 7.62–7.28
(m, 12H), 5.72 (s, 1H), 5.13 (s, 1H), 4.83 (s, 1H), 4.32
(d, 1H, J = 6.2 Hz), 3.53 (m, 2H), 2.68–1.38 (m, 18H),
0.52 (s, 3H); MS (m/e): 669 (MH+), 691 (MNa+);
HRMS: calcd MH+ for C40H39Cl2O3P 669.2092; found
669.2085.
3. Weichert, R.; Neef, G. J. Steroid Biochem. 1987, 27,
851–858.
4. (a) Wehle, H.; Moll, J.; Cato, A. C. B. Steroids 1995, 60,
368–374; (b) Philibert, D.; Costerousse, G.; Gaillard-
Moguilewsky, M.; Nedelec, L.; Nique, F.; Tournemine,
C.; Teutsch, G. Front Human Res. 1991, 19, 1–17.
5. Kehler, J.; Ebert, B.; Dahl, O.; Krogsgarrd-Larsen, P.
Tetrahedron 1999, 55, 771–780.
6. (a) Gange, D.; Magnus, P. J. Am. Chem. Soc. 1978, 100,
7746–7747; (b) Rao, P. N.; wang, Z.; Cessac, J. W.;
Rosenberg, R. S.; Jenkins, D. J. A.; Diamandis, E. P.
Steroids 1998, 63, 523–530.
7. Carroll, F. I.; Tyagi, S.; Blough, B. E.; Kuhar, M. J.;
Navarro, H. A. J. Med. Chem. 2005, 48, 3852–3857.
8. (a) Gibertson, S. R.; Fu, Z.; Starkey, G. W. Tetrahedron
Lett. 1999, 40, 8509–8512; (b) Abbas, S.; Hayes, C. J.
Tetrahedron Lett. 2000, 41, 4513–4517; (c) Ogawa, T.;
Usuki, N.; Ono, N. J. Chem. Soc., Perkin Trans. 1 1998,
17, 2953–2958.
9. Miyaura, N.; Suzuki, A. Chem. Rev. 1995, 95, 2457–2483.
10. (a) Boger, D. L.; Castle, S. L.; Miyazaki, S.; Wu, J. H.;
Richard, R.; Beresis, R. T.; Loiseleur, O. J. Org. Chem.
1999, 64, 70–80; (b) Dziemidowicz, J.; Witt, D.; Sliwka-
Kaszynska, M.; Rachon, J. Synthesis 2005, 569–574; (c)
Hum, G.; Grzyb, J.; Taylor, S. D. J. Comb. Chem. 2000, 2,
234–242.
5.2.16. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-ep-
oxy-11-(4-(diphenoxy-phosphorylphenyl)-, (11b,17a)-
(9CI) (20j). The title compound was prepared as a white
solid (160 mg, 46%), according to the procedure for
compound 20b, starting from compound 19 (300 mg,
1
0.547 mmol). H NMR (CDCl3) d 7.82 (m, 2H), 7.28–
7.08 (m, 12H), 5.72 (s, 1H), 5.14 (s, 1H), 4.82 (s, 1H),
4.32 (d, 1H, J = 5.8 Hz), 3.82 (m, 2H), 2.71–1.41 (m,
18H), 0.51 (s, 3H); MS (m/e): 633 (MH+), 655
(MNa+); HRMS: calcd MH+ for C40H41O5P 633.2770;
found 633.2744.
5.2.17. 19,24-Dinorchola-4,9,20-trien-3-one, 17,23-ep-
oxy-11-[4-(5,5-dimethyl-2-oxo-2k5-[1,3,2]dioxaphosph-
inan-2-yl)-phenyl]-, (11b,17a)-(9CI) (20k). The title
compound was prepared as a white solid (100 mg,
33%), according to the procedure for compound 20b,
1
starting from compound 19 (300 mg, 0.547 mmol). H
NMR (CDCl3) d 7.78 (m, 2H), 7.28 (m, 2H), 5.78 (s,
1H), 5.12 (s, 1H), 4.82 (s, 1H), 4.32 (m, 3H), 3.85 (m,
5H), 2.72–1.35 (m, 17H), 1.18 (s, 3H), 1.02 (s, 3H),
0.55 (s, 3H); MS (m/e): 549 (MH+), 571 (MNa+);
HRMS: calcd MH+ for C33H41O5P 549.2770; found
549.2745.
11. Lundeen, S. G.; Zhang, Z.; Zhu, Y.; Carver, J. M.;
Winneker, R. C. J. Steroid Biochem. Mol. Biol. 2001, 78,
137–143.