Discovery of novel phosphorus-containing steroids as selective glucocorticoid receptor antagonist

Article abstract of DOI:10.1016/j.bmcl.2006.10.003

Mifepristone is a non-selective antagonist of 3-oxosteroid receptors with both abortifacient and anti-diabetic activities. For glucocorticoid receptor (GR) program, we sought an unexplored, synthetically accessible phosphorus-containing steroidal mimetic of mifepristone, suitable for parallel synthesis of analogues. One compound 4a, with high oral bioavailability (59%) in rat, exhibited functional antagonism of GR in oral glucose tolerance test (OGTT). Thus this series of compounds might be potentially useful for the treatment of type II diabetes.

Full text of DOI:10.1016/j.bmcl.2006.10.003

Bioorganic & Medicinal Chemistry Letters 17 (2007) 1471–1474
Discovery of novel phosphorus-containing steroids as selective
glucocorticoid receptor antagonist
Weiqin Jiang,* James J. Fiordeliso, George Allan, 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 29 August 2006; revised 29 September 2006; accepted 3 October 2006
Available online 5 October 2006
Abstract—Mifepristone is a non-selective antagonist of 3-oxosteroid receptors with both abortifacient and anti-diabetic activities.
For glucocorticoid receptor (GR) program, we sought an unexplored, synthetically accessible phosphorus-containing steroidal
mimetic of mifepristone, suitable for parallel synthesis of analogues. One compound 4a, with high oral bioavailability (59%) in
rat, exhibited functional antagonism of GR in oral glucose tolerance test (OGTT). Thus this series of compounds might be poten-
tially useful for the treatment of type II diabetes.
ꢀ 2006 Elsevier Ltd. All rights reserved.
Glucocorticoid receptors (GRs), members of the ste-
roid–thyroid–retinoid superfamily, are soluble, intracel-
lular receptor proteins acting as ligand-regulated
transcription factors controlling specific gene expression
in most mammalian cells.1 Glucocorticoids bind to and
activate GRs, and they play a crucial role in the normal
development and maintenance of basal and stress-relat-
ed homeostasis.2 The steroid nucleus of typical gluco-
corticoids is associated with a rich variety of hormonal
activities and has inspired the development of an assort-
ment of therapeutic agents. Potent and selective GR
antagonists could be used for the possible treatment of
a variety of disorders, including diabetes,3 obesity,4
depression,5 neurodegeneration,6 glaucoma,7 and Cush-
ing’s disease.8
There have been literature reports on using phosphinic
acids as bioisosteres of the carboxylic acid group.10
We envisioned that dialkyl or dialkoxyl phosphonyl
groups can serve as bioisosteres in steroids for either
the carbonyl group on the 11b-aryl group of
ZK112993 (2), Org-33628 (3) or N,N-dimethyl group
in RU-486 (1) (Fig. 1). In the present article, we describe
the synthesis and biological properties of phosphorus-
containing steroidal derivatives 4 and 5. According to
X-ray crystal of RU-486 with GR,11 as a replacement
of N,N-dimethyl group in compound 1 or acetyl group
in compounds 2 and 3, phosphate or phosphinic
mono-acid might provide additional functional groups
in GR binding pocket to increase the interactions
toward GR receptor. This might result in both increase
in potency in GR antagonism and selectivity versus PR
antagonism.12
The potent steroidal GR antagonist 1, mifepristone
(RU-486), is known for its abortifacient effects due to
potent progesterone receptor (PR) antagonism (Fig. 1).9
The chemistry used for target compound 4 is shown in
Scheme 1. Thus, commercially available ethylene delta-
none 6 was functionalized at C17 position by the treat-
ment of allyl, propynyl or phenyl Grignard reagent.
Alcohol 7 was converted to epoxide 8 after exposed to
m-CPBA. Cuperate 1,4-addition to epoxide 8 led to
C-11 phenyl substituted compound 9. Deprotection of
THP functional group under acidic condition and reac-
tion with Tf2NPh under basic condition resulted in phen-
yl triflate 10. Coupling reaction with phosphorus reagent
under microwave irradiation condition furnished target
The goal of our GR antagonist program is to identify a
compound with similar potency and higher selectivity
than mifepristone (RU-486) in terms of its progesterone
receptor antagonism.
Keywords: Phosphorus-containing steroids; Selective glucocorticoid
receptor antagonist.
*
Corresponding author. E-mail: wjiang1@prdus.jnj.com
0960-894X/$ - see front matter ꢀ 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2006.10.003

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W. Jiang et al. / Bioorg. Med. Chem. Lett. 17 (2007) 1471–1474
O
O
Me2N
HO
17
O
HO
11
3
1
3
2
O
O
O
RU-486
ORG-33628
ZK112993
O
P
R2
O
R1
R1
P
R2
OH
R
O
4
5
O
O
Figure 1. Structures of steroids and target compounds.
OH
R
O
OH
R
i
ii
O
O
O
O
a. R = allyl
b. R = propynyl
c. R = Ph
O
O
O
6
8
7
O
O
THPO
iii
R1
OH
R
P
F3C
S
O
R2
OH
R
OH
R
O
iv
vi
a. R = allyl
b. R = propynyl
c. R = Ph
O
9
4
10
O
OH
O
O
Scheme 1. Synthetic route for phosphorus-containing steroids. Reagents and condition: (i) RMgBr, THF, 85%; (ii) m-CPBA, DCM, 35%; (iii) 4-
THPO-PhMgBr, CuCl, THF, 53%; (iv) a—oxalic acid or p-TSA, 75%; b—Tf2NPh, NaH, 60%; (v) Pd(OAc)2, dppb, i-Pr2NEt, HP(O)R1R2,
microwave, 20–70%.
compound 4. The synthesis of compound 5 could be
found in Ref. 13.
T47D IC50 to the A549 IC50 was calculated and is listed
in the column ‘Ratio’, as a measure of the separation of
GR and PR antagonism. The commercial drug mifepri-
stone 1 (RU-486) was tested as a control. Compounds
4a–4f are potent GR antagonists, with 4c, almost as
potent as RU-486. Compounds 4a–4f have similar or
better selectivity (up to 127-fold for 4c) than RU-486
(2.6-fold). The interesting observation is that esters 5a
and 5c are active GR or PR antagonists, while both
GR and PR activities are totally abolished for corre-
sponding monoacids 5b and 5d.14
The compounds were tested for their glucocorticoid
receptor (GR) antagonist activity based on their ability
to inhibit corticoid-induced transcription from a gluco-
corticoid response element (GRE)-linked luciferase
reporter gene in the human lung carcinoma cell line
A549 (Table 1). The compounds were also evaluated
for progesterone receptor (PR) antagonist activity based
on their ability to block progesterone induction of alka-
line phosphatase activity in the human breast cancer cell
line T47D. The IC50 of the compounds from the A549
and T47D assays are listed in Table 1. The ratio of the
In order to assess the in vivo potential of this class of
selective GR modulators, liver microsomal metabolism
Table 1. In vitro biological activities for compound 4a–4e and 5a–5d
Compound
R1
R2
R
A549 Kia (nM)
T47b IC50 (nM)
Ratio
RU-486
4a
4b
1.0
2.6
2.60
76.3
30.8
127
Ph
Ph
Allyl
Allyl
Propynyl
Propynyl
Propynyl
Ph
10.0
10.0
3.0
763
308
381
141
101
275
9.9
EtO
Ph
EtO
Ph
4c
4d
EtO
Me
Ph
EtO
Me
Ph
9.0
15.7
10.1
13.8
0.17
—
4e
10.0a
20.0
56.3
>3000
59.8
>3000
4f
5a
EtO
OH
EtO
OH
EtO
OEt
Me
Me
—
—
5b
5c
>1000
1.6
>1000
—
—
0.027
—
5d
a This value is IC50 (nM).

W. Jiang et al. / Bioorg. Med. Chem. Lett. 17 (2007) 1471–1474
1473
Table 2. Pharmacokinetic data for compound 4a compared with RU-486
Compound Fomulation Route Dose (mg/kg) Cmax (lg/mL) CL (mL/min/kg) Tmax (h) t1/2 (h) AUC (mg/h/mL) F (%)
4a
10% soluto iv
Seasame oil po
2
10
5.290
1.847
7.98
0.080
2.00
0.77
2.20
4.173
12.407
—
59
RU-486
10% soluto iv
Seasame oil po
2
10
2.797
0.030
115
0.080
1.00
4.00
2.80
1.371
1.863
—
31
and rat oral/iv pharmacokinetic data were obtained.
Compound 4a has good level of in vitro metabolic
stability in microsome across different species, as shown
in t1/2: human, >30 min; rat, >30 min; mouse, 28 min;
dog, 21 min; monkey, 8 min; rabbit, 30 min. In mice,
nists. The best compounds demonstrated comparable
potency to RU-486 and better selectivity versus PR than
RU-486. Selected compound showed good efficacy in
mice in lowering plasma glucose level. With the potent
and selective GR antagonists in hands, our future efforts
will be focused on achieving liver selectivity.
the GR modulator 4a had
(t = 0.77 h) and high iv clearance (7.98 mL/min/kg), con-
a short iv half-life
sistent with rapid metabolism, observed for steroids.15
References and notes
Compounds 4a and 4b have good CACO-2 permeability
(2.6 · 10À6 cm/s and 14.5 · 10À6 cm/s, respectively).
Compound 4a has a bioavailability of 59% in mice while
RU-486 was 31% tested side by side (Table 2). Com-
pound 4a did not show hERG binding activity (27.8%
inhibition at 10 lM).
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In summary, we have designed and synthesized a series
of novel phosphorus-containing steroidal GR antago-

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