Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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TETRACYCLIC PROGESTERONE RECEPTOR MODULATOR COMPOUNDS
AND METHODS
Field of the Invention
This invention relates to nonsteroidal tetracyclic compounds that are
modulators (i.e.
agonists and antagonists) of progesterone receptors, and to methods for the
making and use
of such compounds.
Background of the Invention
Progesterone receptor (PR) modulators have been widely used in regulation of
female reproduction systems and in treatment of female hormone dependent
diseases. The
effectiveness of known steroidal PR modulators is often tempered by their
undesired side-
effect profile, particularly during long-term administration. For example, the
effectiveness of
synthetic progestins, such as norgestrel, as female birth control agents must
be weighed
against the increased risk of breast cancer and heart disease to women taking
such agents.
Similarly, the progesterone antagonist, mifepristone (RU486), if administered
for chronic
indications, such as uterine fibroids, endometriosis and certain hormone-
dependent cancers,
could cause homeostatic imbalances in a patient due to its inherent cross-
reactivity as a
glucocorticoid receptor (GR) antagonist. Accordingly, identification of
compounds that
have good specificity for PR, but have reduced or no cross-reactivity for
other steroid
receptors, would be of significant value in the improvement of women's health.
Nonsteroidal molecules that contain a di- or tetrahydroquinoline ring as the
core
pharmacophore have been described as steroid receptor modulator compounds.
{See for
example: "Preparation of Quinolines and Fused Quinolines as Steroid Receptor
Modulators", T. K Jones, M. E. Goldman, C. L. F. Pooley, D. T. Winn, J. P.
Edwards, S. J.
West, C. M. Tegley, L. Zhi, L. G. Hamann, R. L. Davis, L. J. Farmer, PCT Int.
Appl. Pub.
No. WO 96/19458; "Steroid Receptor Modulator Compounds and Methods", T. K
Jones,
D. T. Winn, L. Zhi, L. G. Hamann, C. M. Tegley, C. L. F. Pooley, US Patent No.
5,688,808;
"Steroid Receptor Modulator Compounds and Methods", T. K Jones, M. E. Goldman,
C. L.
F. Pooley, D. T. Winn, J. P. Edwards, S. J. West, C. M. Tegley, L. Zhi, US
Patent No.
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5,688,810; "Steroid Receptor Modulator Compounds and Methods", T. K Jones, C.
M.
Tegley, L. Zhi, J. P. Edwards, S. J. West, US Patent No. 5,693,646; "Steroid
Receptor
Modulator Compounds and Methods", T. K Jones, L. Zhi, C. M. Tegley, D. T.
Winn, L. G.
Harnann, J. P. Edwards, S. J. West, US Patent No. 5,693,647; "Steroid Receptor
Modulator
Compounds and Methods", T. K Jones, L. Zhi, J. P. Edwards, C. M. Tegley, S. J.
West, US
Patent No. 5,696,127; "Steroid Receptor Modulator Compounds and Methods", T. K
Jones, D. T. Winn, M. E. Goldman, L. G. Hamann, L. Zhi, L. J. Farmer, R. L.
Davis, US
Patent No. 5,696,130; "Steroid Receptor Modulator Compounds and Methods", T. K
Jones, M. E. Goldman, C. L. F. Pooley, D. T. Winn, J. P. Edwards, S. J. West,
C. M.
Tegley, L. Zhi, L. G. Hamann, L. J. Farmer, R. L. Davis, US Patent No.
5,696,133.}
Molecules containing a bicyclic heterocycle have been reported as cardiotonic
agents. {See:
"A Novel Class of Cardiotonic Agents: Synthesis and Biological Evaluation of 5-
Substituted
3,6-Dihydrothiadiazin-2-ones with Cyclic AMP Phosphoidesterase Inhibiting and
Myofibrillar Calcium Sensitizing Properties", M.-C. Forest, P. Lahouratate, M.
Martin, G.
Nadler, M. J. Quiniou, R. G. Zimmermann, J. Med. Chem. 35 (1992) 163-172;
"Heteroatom Analogues of Bemoradan: Chemistry and Cardiotonic Activity of 1,4-
Benzothiazinylpyridazinones", D. W. Combs, M. S. Rampulla, J. P. Demers, R.
Falotico, J.
B. Moore, J. Med. Chem., 35 (1992) 172-176.}
Summary of the Invention
The present invention is directed to compounds, pharmaceutical compositions,
and
methods for modulating processes mediated by PR. More particularly, the
invention relates
to nonsteroidal compounds and compositions that are high affinity, high
specificity agonists,
partial agonists (i.e., partial activators and/or tissue-specific activators)
and antagonists for
progesterone receptors. Also provided are methods of making such compounds and
pharmaceutical compositions, as well as critical intermediates used in their
synthesis.
These and various other advantages and features of novelty that characterize
the
invention are pointed out with particularity in the claims annexed hereto and
forming a part
hereof. However, for a better understanding of the invention, its advantages,
and objects
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obtained by its use, reference should be had to the accompanying descriptive
matter, in which
preferred embodiments of the invention are described.
Detailed Description of the Invention
As used herein, the following terms are defined with the following meanings,
unless
explicitly stated otherwise.
The terms alkyl, alkenyl, alkynyl and allyl include optionally substituted
straight-
chain, branched-chain, cyclic, saturated and/or unsaturated structures, and
combinations
thereof.
The term haloalkyl refers to alkyl structures, including straight-chain,
branched-chain,
or cyclic structures, or combinations thereof, that are substituted with one
or more fiuorines,
chlorines, bromines or iodines, or combinations thereof.
The term heteroalkyl includes straight-chain, branched-chain, cyclic,
saturated and/or
unsaturated structures, or combinations thereof, in which one or more skeletal
atoms is
oxygen, nitrogen, sulfur, or combinations thereof.
The term aryl refers to an optionally substituted six-membered aromatic ring.
The term heteroaryl refers to an optionally substituted, aromatic five-
membered
heterocyclic ring containing one or more heteroatoms selected from the group
consisting of
oxygen, nitrogen and sulfi~r, or to an optionally substituted, aromatic six-
membered
heterocyclic ring containing one or more nitrogens.
The substituents of an "optionally substituted" structure include, but are not
limited
to, one or more of the following preferred substitutents: F, Cl, Br, I, CN,
NOa, NHz, NCH3,
OH, OCH3, OCF3, CH3, CF3.
Compounds of the present invention are defined as those having the formula:
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(I)
OR
(II)
wherein:
R' through R6 are independently hydrogen, F, Cl, Br, I, N02, CN, OR'°,
NR'°R",
SR'°, COR'z, COzRIZ, CONR'°R", optionally substituted C1 to C6
alkyl or heteroalkyl, C1
to C6 haloalkyl, optionally substituted C3 to Cg cycloalkyl, optionally
substituted C2 to C6
alkenyl or alkynyl, optionally substituted allyl, optionally substituted aryl
or heteroaryl, or
optionally substituted arylmethyl, where R'° and R" are independently
hydrogen, C1 to C6
alkyl or heteroalkyl or haloalkyl, aryl, heteroaryl, optionally substituted
allyl, optionally
substituted arylmethyl, COR'3, S02R'3 or S(O)R'3, where R'z is hydrogen, C1 to
C6 alkyl or
heteroalkyl or haloalkyl, aryl, heteroaryl, optionally substituted allyl or
optionally substituted
arylmethyl, where R'3 is hydrogen, C1 to C6 alkyl or haloallcyl, aryl,
heteroaryl, optionally
substituted allyl or optionally substituted arylmethyl;
R~
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R' is hydrogen, C1 to C6 alkyl or haloallcyl or heteroallcyl, aryl,
arylmethyl,
heteroaryl, COR'z, COzR'z, SOzR'z, S(O)R'z or CONK'°R", where
R'°''z have the same
definitions given above;
Rg and R9 are independently hydrogen, C1 to C6 allcyl or haloalkyl or
heteroalkyl,
optionally substituted C2 to C6 alkenyl or alkynyl, optionally substituted
allyl, optionally
substituted arylinethyl, optionally substituted aryl or optionally substituted
heteroaryl;.
X is OCHz, SCHz, NHCHz, OC(O), SC(O), NHC(O), CH20, CHzS, CHzNH,
C(O)O, C(O)S or C(O)NH;
Y is O, S or NR'°, where R'° has the same definition given
above; and
Z is O, S, NR'4, CR'4R's, CR'4R'sCR'6R", OCR'4RIS, SCR'4R's, CR'4R'sS,
NR'4CR'sR'6, or CR'4R'sNR'6, where R'4 through R" each independently are
hydrogen, CI
to C6 alkyl or haloallcyl or heteroalkyl, optionally substituted C2 to C6
alkenyl
or alkynyl, optionally substituted allyl, optionally substituted arylmethyl,
optionally
substituted aryl or optionally substituted heteroaryl;
or a pharmaceutically acceptable salt thereof .
In a preferred aspect, the present invention provides a pharmaceutical
composition
comprising an effective amount of an progesterone receptor modulating compound
of
formula I or formula II shown above wherein R'-", X, Y and Z all have the same
definitions
as given above.
In a fiirther preferred aspect, the present invention comprises a method of
modulating
processes mediated by progesterone receptors comprising administering to a
patient an
effective amount of a compound of formula I or formula II shown above, wherein
R'-", X,
Y and Z all have the same definitions as those given above.
Any of the compounds of the present invention can be synthesized as
pharmaceutically acceptable salts for incorporation into various
pharmaceutical
compositions. As used herein, pharmaceutically acceptable salts include, but
are not limited
to, hydrochloric, hydrobromic, hydroiodic, hydrofluoric, sulfuric, citric,
malefic, acetic, lactic,
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nicotinic, succinic, oxalic, phosphoric, malonic, salicylic, phenylacetic,
stearic, pyridine,
ammonium, piperazine, diethylamine, nicotinamide, formic, urea, sodium,
potassium,
calcium, magnesium, zinc, lithium, cinnamic, methylamino, methanesulfonic,
picric, tartaric,
triethylamino, dimethylamino, and tris(hydroxymethyl)aminomethane. Additional
pharmaceutically acceptable salts are known to those skilled in the art.
The PR agonist, partial agonist and antagonist compounds of the present
invention
are particularly useful for female hormone replacement therapy and as
modulators of fertility
(e.g., as contraceptives or contragestational agents), either alone or in
conjunction with
estrogen receptor modulators. The PR modulator compounds are also used in the
treatment
of dysfunctional uterine bleeding, dysmenorrhea, endometriosis, leiomyomas
(uterine
fibroids), hot flashes, mood disorders, meningiomas as well as in various
hormone-dependent
cancers, including, without limitation, cancers of ovaries, breast,
endometrium and prostate.
It will be understood by those skilled in the art that while the compounds of
the
present invention will typically be employed as a selective agonists, partial
agonists or
antagonists, that there may be instances where a compound with a mixed steroid
receptor
profile is preferred. For example, use of a PR agonist (e.g., progestin) in
female
contraception often leads to the undesired effects of increased water
retention and acne flare-
ups. In this instance, a compound that is primarily a PR agonist, but also
displays some AR
and MR modulating activity, may prove useful. Specifically, the mixed MR
effects would be
useful to control water balance in the body, while the AR effects would help
to control any
acne flare-ups that occur.
Furthermore, it will be understood by those skilled in the art that the
compounds of
the present invention, including pharmaceutical compositions and formulations
containing
these compounds, can be used in a wide variety of combination therapies to
treat the
conditions and diseases described above. Thus, the compounds of the present
invention can
be used in combination with other hormones and other therapies, including,
without
limitation, chemotherapeutic agents such as cytostatic and cytotoxic agents,
immunological
modifiers such as interferons, interleukins, growth hormones and other
cytokines, hormone
therapies, surgery and radiation therapy.
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Representative PR modulator compounds (i.e., agonists, partial agonists and
antagonists) according to the present invention include: 7-ffuoro-4,4-dimethyl-
SH-
chromeno[3,4 f]-1,3-benzo[d]oxazin-2-one (Compound 14); 9-bromo-7-ffuoro-4,4-
dimethyl-SH chromeno[3,4 f]-1,3-benzo[d]oxazin-2-one (Compound 20); 7-ffuoro-9-
forrrryl-4,4-dimethyl-SH-chromeno[3,4 f]-1,3-benzo[djoxazin-2-one (Compound
24); 7-
ffuoro-9-hydroxyiminomethyl-4,4-dimethyl-SH chromeno[3,4 fj-1,3-benzo[d]oxazin-
2-one
(Compound 25); 9-cyano-7-ffuoro-4,4-dimethyl-SH-chromeno[3,4 f]-1,3-
benzo[d]oxazin-2-
one (Compound 26).
Compounds of the present invention, comprising classes of heterocyclic
nitrogen
compounds and their derivatives, can be obtained by routine chemical synthesis
by those
skilled in the art, for example, by modification of the heterocyclic nitrogen
compounds
disclosed or by a total synthesis approach.
The sequences of steps to synthesize the compounds of the present invention
are
shown below in the general schemes. In each of the Schemes the R groups (e.g.,
R', R2,
etc.) correspond to the specific substitution patterns noted in the Examples.
However, it will
be understood by those skilled in the art that other functionalities disclosed
herein at the
indicated positions of compounds of formulas I and II also comprise potential
substituents
for the analogous positions on the structures within the Schemes. In a further
aspect, the
present invention provides a novel process for the preparation of the
compounds of the
present invention.
Scheme I
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R~ COZR R1
R Br 2
RZ F 1 n-BULI R2 F ~ R / F
2~ B(OMe~ I coZR
\ 3) HsO+ RS \ NOz Rs \
R3 H ~ R3 \ B~OH)2 Rs
RS Noz
Rs
Pd coupling
R1
1) Pd/C, Hz Rz / 0 0
1 ~ NaOH DMF
2 NaH, 2)~ 3 \ ~ Br
DIGtF"'~ R
4
NOZ R5 NHZ
6 Rs
1) Pd(PP~a 1) LAH
RB(OH~ 2)Et3SiH, TFA
2) ClC4Me
DMAP
OZMe 02Me
DCE, TsOH
Rs
The process of Scheme I begins with the preparation of fiuoroboronic acid 2 by
a
literature procedure, in which ortho-lithiation of ffuorobenzene 1 with an
alkyl lithium
reagent, such as n-butyllithium (n-BuLi) or tert-butyllithium, in THF or ether
at -78 to -50 °C
followed by addition of a trialkyl borate, such as trimethyl or triisopropyl
borate, at -78 °C
and acidification with an aqueous acid, such as HCI, provides boronic acid 2.
Biaryl
compound 4 is prepared by a typical palladium catalyzed coupling reaction of
boronic acid 2
and bromobenzoate 3 under aqueous or nonaqueous conditions at ambient
temperature. The
ester 4 is hydrolyzed under basic conditions, such as THF/MeOH/2N NazC03, at
ambient
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temperature, and the resulting carboxylate is heated at elevated temperature
in a polar
solvent, such as DMF, to generate lactone 5. The nitro-lactone 5 is converted
to
aminobromo-lactone 6 in a one-pot two-step procedure that involves reduction
of the nitro
group to an amino group under hydrogen atmosphere (catalyzed by palladium on
carbon)
followed by bromination with N-bromosuccinimide (NBS) in DMF at room
temperature.
The isopropenyl group is introduced by a palladium catalyzed coupling
reaction, for
example, Suzuki coupling reaction between isopropenylboronic acid and bromo
compound
6. The resulting amino compound is then converted to the carbamate 7 by
treatment with
methyl chloroformate in the presence of 4-dimethylaminopyridine. Removal of
the carbonyl
group of lactone 7 is completed by stepwise reduction with typical reducing
agents such as
LiAlH4 and Et3SiH in the presence of a catalytic amount of acid (e.g., TFA) to
afford
compound 8. The final product 9 is obtained by the treatment of compound 8
with tosic
acid (TsOH, p-toluenesulfonic acid) in refiuxing dichloroethane (DCE).
Scheme II
H _0H
LiBHq, THF Rs ~ /
4
OZMe R RS~NHC4zMe
i Rs 10
I) NBS, Et3N
2) MeI, Na2C03 DCE, TsOH
3) NaH, DMF
'11 R6
Oz Me
Scheme II describes a four-step, selective D-ring modification procedure, in
which
reduction of lactone 7 with a reducing agent such as LiBH4 provides diol 10
and then NBS
bromination of diol 10 in the presence of a base such as triethylamine
followed by a selective
methylation and NaH mediated nucleophilic cyclization in DMF affords compound
11.
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Treatment of compound 11 with more than one equivalent of an acid such as TsOH
in
refluxing dichloroethane provides compound 12.
Scheme III involves selective D-ring fiznctional group conversion from R'-4 to
R'-'°
by known substituent group conversions such as converting bromo to aldehyde by
metal-
s halogen exchange followed by nucleophilic addition to DMF, or converting an
aldehyde to
an oxime by hydroxylamine treatment of the aldehyde, or converting an oxime to
a cyano
group by treatment of the oxime with thionyl chloride.
Scheme III
D-ring
1110dIfICati~ R ~ ~ O~ Me~Me
R9
Rio ~ i
R5~ H
g Rs ' ' 12 Rs
Scheme IV
Rt
Lawesson's
reagent
--~ R'
Scheme IV describes the conversion of compound 12 to its cyclic thiocarbamate
analogue 13 by Lawesson's reagent (p-methoxyphenylthionophosphine sulfide
dimer).
It will be understood by those skilled in the art that certain modifications
can be made
to the above-described methods that remain within the scope of the present
invention.
The compounds of the present invention also include racemates, stereoisomers
and
mixtures of said compounds, including isotopically-labeled and radio-labeled
compounds.
h 3 Rs . .
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Such isomers can be isolated by standard resolution techniques, including
fractional
crystallization and chiral column chromatography.
As noted above, any of the PR modulator compounds of the present invention can
be
combined in a mixture with a pharmaceutically acceptable ca.rner to provide
pharmaceutical
compositions useful for treating the biological conditions or disorders noted
herein in
mammalian, and more preferably, in human patients. The particular carrier
employed in
these pharmaceutical compositions may take a wide variety of forms depending
upon the
type of administration desired, e.g., intravenous, oral, topical, suppository
or parenteral.
In preparing the compositions in oral liquid dosage forms (e.g., suspensions,
elixirs
and solutions), typical pharmaceutical media, such as water, glycols, oils,
alcohols, flavoring
agents, preservatives, coloring agents and the like can be employed.
Similarly, when
preparing oral solid dosage forms (e.g., powders, tablets and capsules),
carriers such as
starches, sugars, diluents, granulating agents, lubricants, binders,
disintegrating agents and
the like will be employed. Due to their ease of administration, tablets and
capsules represent
the most advantageous oral dosage form for the pharmaceutical compositions of
the present
invention.
For parenteral administration, the Garner will typically comprise sterile
water,
although other ingredients that aid in solubility or serve as preservatives
may also be
included. Furthermore, injectable suspensions may also be prepared, in which
case
appropriate liquid carriers, suspending agents and the like will be employed.
For topical administration, the compounds of the present invention may be
formulated
using bland, moisturizing bases, such as ointments or creams. Examples of
suitable ointment
bases are petrolatum, petrolatum plus volatile silicones, lanolin, and water
in oil emulsions such
as EucerinTM (Beiersdorf). Examples of suitable cream bases are NiveaTM Cream
(Beiersdorf),
cold cream (USP), Purpose CreamTM (Johnson & Johnson), hydrophilic ointment
(USP), and
LubridermTM (Warner-Lambert).
The pharmaceutical compositions and compounds of the present invention will
generally be administered in the form of a dosage unit (e.g., tablet, capsule
etc.) at from
about 1 ~g/kg of body weight to about 500 mg/kg of body weight, more
preferably from
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about 10 ~g/kg to about 250 mg/kg, and most preferably from about 20 ~g/kg to
about 100
mg/kg. As recognized by those skilled in the art, the particular quantity of
pharmaceutical
composition according to the present invention administered to a patient will
depend upon a
number of factors, including, without limitation, the biological activity
desired, the condition
of the patient, and tolerance for the drug.
The compounds of this invention also have utility when radio- or isotopically-
labeled
as ligands for use in assays to determine the presence of PR in a cell
background or extract.
They are particularly useful due to their ability to selectively activate
progesterone receptors,
and can therefore be used to determine the presence of such receptors in the
presence of
other steroid receptors or related intracellular receptors.
Due to the selective specificity of the compounds of this invention for
steroid
receptors, these compounds can be used to purify samples of steroid receptors
in vitro. Such
purification can be carried out by mixing samples containing steroid receptors
with one or
more of the compounds of the present invention so that the compounds bind to
the receptors
of choice, and then separating out the bound ligand/receptor combination by
separation
techniques that are known to those of skill in the a.rt. These techniques
include column
separation, filtration, centrifugation, tagging and physical separation, and
antibody
complexing, among others.
The compounds and pharmaceutical compositions of the present invention possess
a
number of advantages over previously identified steroid modulator compounds.
For
example, the compounds are extremely potent activators of PR, preferably
displaying 50%
maximal activation of PR at a concentration of less than 100 nM, more
preferably at a
concentration of less than 50 nM, more preferably yet at a concentration of
less than 20 nM
or less. Also, the selective compounds of the present invention generally do
not display
undesired cross-reactivity with other steroid receptors, as is seen with the
compound
mifepristone (RU486; Roussel Uclaf), a known PR antagonist that displays an
undesirable
cross reactivity on GR, thereby limiting its use in long-term, chronic
administration. In
addition, the compounds of the present invention, as small organic molecules,
are easier to
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synthesize, provide greater stability and can be more easily administered in
oral dosage forms
than other known steroidal compounds.
The invention will be fiirther illustrated by reference to the following non-
limiting
Examples.
EXAMPLE 1
7-Fluoro-4,4-dimethyl-5H-chromenof3,4-f1-1,3-benzoldloxazin-2-one (Comuound
14;
Structure 9 of Scheme I, where R' = tluoro, RZ~ = H)
2 3-Difluoroboronic acid (Structure 2 of Scheme I. where R' = fluoro. R2-4 =
H)
To a 500 mL flask charged with a solution of 1,2-difluorobenzene (15 g, 0.13
mmol)
in THF (150 mL) at -78 °C was added dropwise n-BuLi (7.0 M in hexane,
19 mL, 0.13 mol).
The reaction mixture was stirred at -78 °C for 2.5 hours and then a
solution of
trimethylborate (30 mL, 0.26 mol, 2.0 equiv) in THF (90 mL) previously cooled
to - 78 °C
was added, after which the mixture was allowed to warm up to room temperature
overnight.
Next the reaction mixture was acidified to pH ~ 1 using HCl (3 N aqueous) and
then stirred
for 15 minutes. Then the mixture was extracted with ether (2 x 300 mL), washed
with water
(150 mL) and dried over Na2S04. The solvent was removed in vacuo to yield 20 g
(96%) of
2,3-difluoroboronic acid as a white solid. Data for 2,3-difluoroboronic acid:
rf = 0.39
(EtOAc:hexanes, 3:7).
Methyl 2-(2 3-Difluorophenyl)-5-nitrobenzoate (Compound 15, Structure 4 of
Scheme I.
where R' = fluoro. R2~6 = H)
A mixture of 2,3-difluoroboronic acid (20g, 0.12 mol, 1.3 equiv), methyl 2-
bromo-5-
2~ nitrobenzoate (25 g, 96 mmol) (Structure 3, where R = methyl, RS-6 = H),
tetrakis(triphenylphosphine)palladium(0) (3.6 g, 3.1 mmol, 3.2% equiv) and
aqueous sodium
carbonate (2 M, 200 mL) in toluene (200 mL) and ethanol (100 mL) was heated at
reflux
overnight until completion of the reaction was indicated by TLC. The reaction
mixture was
extracted with EtOAc (2 x 400 mL), washed with brine (300 mL) and dried over
NazS04.
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Removal of solvent provided a brown solid, which was recrystallized from i-
PrOH:hexanes
to give Compound 15 (22.8 g, 83%) as a white solid. Data for 15: rf= 0.32
(CHzClz:hexanes, 4:6); 'H NMR (400 MHz, CDC13) 8 8.87 (d, J= 1.2, 1 H), 8.43
(dd, J=
8.3 and 1.2, 1 H), 7.58 (d, J= 8.6, 1 H), 7.29-7.17 (m, 2 H), 7.10-7.02 (m, 1
H) and 3.82 (s,
3 H).
4-Fluoro-8-vitro-6H dibenzo[b,d]pyran-6-one (Compound 16: Structure 5 of
Scheme I,
where R' = fluoro. R2-6 = H)
To a solution of Compound 15 (20 g, 69 mmol) in THF:MeOH 02.5:1) (370 mL)
was added 10% aqueous NaOH (82 mL), and the reaction mixture was stirred at
room
temperature for 1 hour. The mixture was concentrated, acidified to pH ~ 1
using 3N HCl
and then extracted with EtOAc (2 x 400 mL). The combined extracts were washed
with
brine, dried over NazS04 and concentrated to afford the acid as an off white
solid. To a
solution of the crude acid in dry DMF (180 mL) was added NaH (4.0 g, 1.5
equiv), and the
mixture was heated at ~80 °C for 2 hours, at which time TLC indicated
completion of the
reaction. The reaction mixture was concentrated under reduced pressure to a
small volume,
and then water (5 mL) was added. The mixture was cooled to -15 °C to
afford a white
precipitate, which was filtered and washed with cold water and hexane to give
Compound 16
(17 g, 95%). Data for 16: rf= 0.36 (EtOAc:hexanes, 3:7); 'H NMR (400 MHz,
CDC13) 8
9.28 (d, J= 1.1, 1 H), 8.68 (dd, J= 8.5 and 1.1, 1 H), 8.32 (d, J= 8.5, 1 H),
7.91 (d, J=
8.0, 1 H) and 7.46-7.36 (m, 2 H).
8-Amino-7-bromo-4-fluoro-6H dibenzofb,dlnvran-6-one (Compound 17: Structure 6
of
Scheme I. where R' = ffuoro. R2-6 = H)
A mixture of Compound 16 (8.8 g, 34 mmol) and 10% Pd/C (0.95 g, 2.5% equiv.)
in
DMF (150 mL) in a Parr apparatus was shaken at room temperature under HZ (40-
60 psi)
overnight until completion of the hydrogenation was indicated by TLC. The
reaction
mixture was then carefully passed through filter paper to remove all traces of
Pd/C catalyst.
NBS (6.0 g, 34 mmol) was added to the filtrate, and the resulting mixture was
stirred at
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room temperature for 2-3 hours. The mixture was concentrated under reduced
pressure, and
then water was added to initiate precipitation. The solid was filtered and
washed with cold
water to afford Compound 17 (8.6 g, 83%) as a pale brown solid. Data for 17:
rf = 0.11
(EtOAc:hexanes, 1:3); 'H NMR (400 MHz, DMSO-d6) 8 8.20 (d, J= 8.8, 1 H), 7.97
(d, J=
8.0, 1 H), 7.38 (d, J= 8.8, 1 H), 7.35-7.27 (m, 2 H), 6.24 ( s, 2 H).
4-Fluoro-7-isopro~envl-8-methox~carbonylamino-6H-dibenzo[b,djpyran-6-one
(Compound
18; Structure 7 of Scheme I, where R' = ffuoro, Rz-6 = H)
To a solution of 2-bromopropene (1.2 g, 10 mmol) in THF (25 mL) at -78
°C was
added t-BuLi (1.7 M in pentane, 12 mL, 20 mmol), and the resulting yellow
solution was
stirred for 10 minutes. Trimethylborate (3.0 mL, 26 mmol) was added via
syringe, and the
reaction mixture was warmed up slowly overnight to yield a white slurry. The
reaction
mixture was acidified to pH ~ 2, stirred at room temperature for 1 hour,
extracted with
EtOAc (2 x 50 mL), washed with brine and then was concentrated to afford the
crude
boronic acid as a white solid.
A mixture of this crude boronic acid, Compound 17 (1.5 g, 4.8 mmol), KZC03
(2.8 g,
mmol) and Pd(PPh3)4 (0.10 g, 0.087 mmol, 1.8% equiv) in toluene (45 mL), EtOH
(45
mL) and water (20 mL) was heated at reflex for 2 hours. The dark reaction
mixture was
acidified to pH ~ 2 and extracted with EtOAc (2 x 150 mL). Removal of solvent
provided a
20 crude dark solid that contained 8-amino-4-ffuoro-7-isopropenyl-6H
dibenzo[b,d]pyran-6-
one. To the crude mixture in THF (40 mL) at room temperature was added CICOzMe
(1.7
mL, 20 mmol) and DMAP (0.53 g, 4.3 mmol), and the resulting cloudy solution
was stirred
at room temperature for 5 hours. The reaction was quenched with water (50 mL),
and the
reaction mixture was extracted with EtOAc and washed with aqueous Na2C03,
NH4C1 and
brine. Removal of solvent and chromatography of the crude mixture afforded
Compound 18
(0.35 g, 26%) as a pale yellow solid. Data for 18: rf = 0.34 (EtOAc:Hexane,
1:3); 'H NMR
(400 MHz, CDCI3) 8 8.72 (d, J= 8.9, 1 H), 8.08 (d, J= 9.0, 1 H), 7.79 (bd, J=
6.5, 1 H),
7.54 (s, 1 H), 7.24-7.18 (m, 2 H), 5.52 (s, 1 H), 4.95 (s, 1 H), 3.81 (s, 3 H)
and 2.14 (s, 3
H).
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4-Fluoro-7-isopropenyl-8-methoxycarbonylamino-6H-dibenzo[b,dipyran (Compound
19:
Structure 8 of Scheme I. where R' = ffuoro, Rz-6 = H)
To a solution of Compound 18 (70 mg, 0.21 mmol) in THF (8 mL) was added
LiAlH4 (8.0 mg, 0.21 mmol), and the resulting mixture was stirred at room
temperature for
30 minutes. The reaction was quenched with water, and the reaction mixture was
extracted
with EtOAc and concentrated. Chromatography afforded 4-fluoro-6-hydroxy-7-
isopropenyl-
8-methoxycarbonylamino-6H dibenzo[b,d]pyran (20 mg, 28%) as a yellow solid,
which was
treated with a catalytic amount of TFA in the presence of Et3SiH (0.2 mL) and
CHZCIz (4
mL) for 2 hours at room temperature. Purification provided Compound 19 (13 mg,
68%) as
a solid. Data for 19: 'H NMR (400 MHz, CDCl3) 8 8.13 (d, J= 8.2, 1 H), 7.61
(d, J= 8.6,
1 H), 7.46 (d, J= 7.5, 1 H), 7.04-6.93 (m, 2 H), 6.89 (s, 1 H), 5.55 (s, 1 H),
5.10 (d, J=
13.5, 1 H), 5.07 (d, J= 13.5, 1 H), 5.03 (s, 1 H), 3.79 (s, 3 H) and 2.01 (s,
3 H).
7-Fluoro-4,4-dimethXl-SH chromeno[3,4 f]-1,3-benzofdJoxazin-2-one (Compound
14;
Structure 9 of Scheme I. where R' = ffuoroyRz-6 = H)
A mixture of Compound 19 (13 mg, 0.041 mmol) and TsOH (16 mg, 0.084 mmol) in
dichloroethane (5 mL) was heated at reflux for 15 hours and concentrated. The
mixture was
diluted in EtOAc (20 mL) and was washed with aqueous Na2C03 (2 x 5 mL) and
brine.
Removal of solvent provided the product as a white solid, which was
recrysta.llized from
EtOAc:hexanes to yield 6 mg (49%) of Compound 14 as a white solid. Data for
14: rf =
0.23 (EtOAc:hexanes, 1:l); 'H NMR (400 MHz, CDC13) 8 8.71 (bs, 1 H), 7.58 (d,
J= 8.4,
1 H), 7.40 (d, J= 7.4, 1 H), 7.08-6.95 (m, 2 H), 6.88 (d, J= 8.4, 1 H), 5.24
(s, 2 H) and
1.84 (s, 6 H).
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FXAMP1.F 7
9-B romo-7-fluoro-4,4-dimethyl-SH-chromeno f 3,4-fl-1,3-benzo f dl oxazin-2-
one
(Compound 20; Structure 12 of Scheme II, where R' = fluoro, R9 = bromo,
Rs-s=Rs=Rio=I~
N-Methoxycarbonyl-3-hydroxymethyl-4-(3-fluoro-2-hydroxYphenyl)-2-
isopropenylaniline
(Compound 21: Structure 10 of Scheme II, where R' = fluoro, Rz-6 = H)
To a solution of Compound 18 (Structure 7 of Scheme II, where R' = ffuoro, Rz-
6 -
H) (0.33 g, 1.0 mmol) in THF (30 mL) was added LiAlH4 (44 mg, 2.0 mmol)
portionwise at
room temperature, and the resulting mixture was stirred at room temperature
for 2 hours.
The reaction was quenched with water, and the reaction mixture was extracted
with EtOAc.
Removal of solvent followed by chromatography afforded Compound 21 (0.30 g,
96%) as a
colorless oil. Data for 21: rf= 0.11 (EtOAc:hexanes, 1:3); 'H NMR (400 MHz,
CDC13)
showed a mixture of rotomers.
N-Methoxycarbonyl-3-h d~xymethyl-4-(5-bromo-3-ffuoro-2-h d~xyphenyl)-2-
isopropenylaniline (Compound 22: Structure 10 of Scheme II, where R' = fluoro.
R3 =
bromo. Rz = R4-6 = H)
NBS (0.18 g, 1.0 mmol) was added to a mixture of Compound 21 (0.30 g, 0.96
mmol) and Et3N (1.0 mL) in CHzCIz (12 mL) at room temperature. After 10
minutes, the
mixture was diluted with EtOAc (SO mL), and washed with water, aqueous NH4Cl
and brine.
Removal of solvent and chromatography of the residue provided 0.34 g (86%) of
Compound
22 as a yellow oil. Data for 22: rf = 0.12 (EtOAc:hexanes, 1:3).
2-Bromo-4-ffuoro-7-isopropenyl-8-methox~carbonylamino-6H-dibenzo[b-dd~pyran
(Compound 23: Structure 11 of Scheme II. where R' = ffuoro. R9 = bromo, RS-6 =
R8 = R'°
To a solution of Compound 22 (0.34 g, 0.83 mmol) in DMF (10 mL) was added
KzC03 (0.14 g, 1.0 mmol) and MeI (0.5 mL, excess), and the mixture was stirred
at room
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temperature for 1 hour. Standard work-up followed by chromatography afforded
0.28 g
(78%) ofN-methoxycarbonyl-2-isopropenyl-3-hydroxymethyl-4-(5-bromo-3-ffuoro-2-
methoxyphenyl)aniline. Data for the methylated intermediate: rf = 0.52
(EtOAc:hexanes,
1:1); 'H NMR (400 MHz, CDCl3) (rotomers) 8 8.13/8.02 (bs, 1 H), 7.30 (m, 1 H),
7.17-
6.95 (m, 3 H), 5.61/5.53 (s, 1 H), 5.21/4.97 (s, 1 H), 4.50-4.12 (m, 2 H),
3.79/2.95 (s, 3 H),
3.62/2.88 (s, 3 H) and 2.20/2.02 (s, 3 H).
A mixture of the methylated intermediate (0.28 g, 0.65 mmol) and NaH (30 mg,
0.75
mmol) in DMF (10 mL) was heated in an 80 °C oil bath for 2 hours until
the reaction went to
completion. Standard work-up followed by chromatography afforded 0.20 g (80%)
of
Compound 23 as a solid. Data for 23: rf= 0.65 (EtOAc:hexanes, 1:1); 'H NMR
(400
MHz, CDCl3) 8 8.15 (d, J= 8.7, 1 H), 7.58 (m, 1 H), 7.56 (d, J= 8.7, 1 H),
7.15 (dd, J=
9.5 and 2.0, 1 H), 6.89 (s, 1 H), 5.56 (s, 1 H), 5.09 (d, J= 12, 1 H), 5.07
(d, J= 12, 1 H),
5.03 (s, 1 H), 3.79 (s, 3 H) and 2.00 (s, 3 H).
9-Bromo-7-ffuoro-4.4-dimethyl-5H chromeno[3.4 f]-1,3-benzo[d]oxazin-2-one
(Compound
20; Structure 12 of Scheme II, where R' = fluoro. R9 = bromo, R5~6 = R8 =
R'° = H)
A mixture of Compound 23 (0.12 g, 0.31 mmol) and TsOH (0.12 g, 0.62 mmol) in
dichloroethane (15 mL) was heated at reflux for 15 hours and concentrated. The
mixture
was diluted in EtOAc (50 mL) and then washed with aqueous Na2C03 (2 x 10 mL)
and
brine. Removal of solvent provided the product as a white solid, which was
recrystallized
from EtOAc:hexanes to give 60 mg (49%) of Compound 20. Data for 20: rf = 0.30
(EtOAc:hexanes, 1:1); 'H NMR (400 MHz, CDCl3) 8 8.09 (s, 1 H), 7.53 (d, J=
8.3, 1 H),
7.52 (m, 1 H), 7.19 (dd, J= 9.4 and 1.9, 1 H), 5.22 (s, 2 H) and 1.82 (s, 6
H). Some
starting material was also recovered (55 mg, 45%).
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EXAMPLE 3
7-Fluoro-9-formvl-4,4-dimethvl-5H-chromeno f 3,4-fl-1,3-benzo f dl oxazin-2-
one
(Compound 24; Structure 12 of Scheme III, where R' = fluoro, R9 = formyl,
Rs-s=Rs=Rio=I~
MeLi (1.4 M in ether, 0.10 mL, 0.14 mmol) was added to a -70 °C
solution of
Compound 20 (50 mg, 0.13 mmol) in THF (12 mL), and the resulting mixture was
stirred for
minutes before n-BuLi (1.6 M in hexane, 0.10 mL, 0.16 mmol) was added. The
reaction
mixture was warmed up to -40 °C and then cooled back down to -70
°C. DMF (0.40 mL,
5.0 mmol) was added to the reaction mixture, which was then warmed to room
temperature,
10 quenched with water (10 mL) and extracted with EtOAc (2 x 20 mL).
Chromatography
afforded 26 mg (61 %) of Compound 24 as a white solid. Data for 24: rf = 0.13
(EtOAc:hexanes, 1:1); 'H NMR (400 MHz, CDC13) 8 9.97 (d, J= 1.8, 1 H), 9.33
(bs, 1 H),
8.19 (t, J= 1.3, 1 H), 7.90 (d, J= 8.4, 1 H), 7.61 (dd, J= 10.3 and 1.3, 1 H),
7.15 (d, J=
8.4, 1 H), 5.51 (s, 2 H) and 1.83 (s, 6 H).
EXAMPLE 4
7-Fluoro-9-hvdroxviminomethvl-4,4-dimethvl-5H-.chromenol3,4-fl-1,3-benzof
d)oxazin-
2-one (Compound 25; Structure 12 of Scheme III, where R' = fluoro, R9 =
hydroxyiminomethyl, Rs-6 = Rs = Rlo = H)
NHZOH-HCl (10 mg, 0.14 mmol) and pyridine (0.1 mL, 1.4 mmol) were added to a
solution of Compound 24 (20 mg, 0.061 mmol) in ethanol (4 mL), and the
resulting mixture
was stirred at room temperature for 1 hour. The reaction mixture was
concentrated,
dissolved in EtOAc (30 mL), washed with water and brine, and re-concentrated
to afford 18
mg (86%) of Compound 25 as a white solid. Data for 25: rf = 0.11
(EtOAc:hexanes, l : l);
'H NMR (400 MHz, acetone-d6) 8 9.27 (s, 1 H), 8.15 (s, 1 H), 7.80 (s, 1 H),
7.78 (d, J=
8.3, 1 H), 9.39 (d, J= 11.2, 1 H), 7.11 (d, J= 8.3, 1 H), 5.39 (s, 2 H) and
1.81 (s, 6 H).
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EXAMPLE 5
9-Cyano-7-fluoro-4,4-dimethyl-5H-chromeno f 3,4-fl-1,3-benzo f d1 ozazin-2-one
(Compound 26; Structure 12 of Scheme IH, where R' = fluoro, R9 = cyano, R5~ =
R8 =
R'°H)
Compound 25 (10 mg, 0.029 mmol) was treated with thionyl chloride (0.032 mL,
0.043 mmol) in dichloromethane (10 mL) at room temperature for 40 minutes. The
reaction
mixture was then quenched with a saturated Na2C03 solution (2 mL), extracted
with EtOAc
(2 x 20 mL) and washed with brine. Removal of solvent followed by
chromatography
afforded 8 mg (90%) of Compound 26 as a white solid. Data for 26: rf = 0.32
(EtOAc:hexanes, 1:1); 'H NMR (400 MHz, acetone-d6) 8 9.35 (bs, 1 H), 8.05 (t,
J= 1.5, 1
H), 7.90 (d, J= 8.4, 1 H), 7.59 (dd, J= 10.6 and 1.5, 1 H), 7.15 (d, J= 8.4, 1
H), 5.51 (s, 2
H) and 1.82 (s, 6 H).
Steroid Receptor Activity
Utilizing the cotransfection assay described by R. M. Evans, Science, 240
(1988)
889-895, the disclosure of which is herein incorporated by reference, the
compounds of the
present invention were tested and found to have strong, specific activity as
agonists, partial
agonists and antagonists of PR. This assay is described in further detail in
the following U. S.
Patents, the disclosures of which are incorporated herein by reference:
"Retinoic Acid
Receptor Method", R. M. Evans, E. Ong, P. S. Segui, C. C. Thompson, K Umesono,
V.
Giguere, US Patent No. 4,981,784; and "Hormone Receptor-Related Assays", R. M.
Evans,
C. A. Weinberger, S. M. Hollenberg, V. Giguere, J. Arriza, C. C. T'hompson, E.
S. Ong, US
Patent No. 5,071,773.
The cotransfection assay provides a method for identifying functional agonists
and
partial agonists that mimic, or antagonists that inhibit, the effect of native
hormones, and for
quantifying their activity for responsive intracellular receptor (IR)
proteins. In this regard,
the cotransfection assay mimics an in vivo system in the laboratory.
Importantly, activity in
the cotransfection assay correlates very well with known in vivo activity,
such that the
cotransfection assay functions as a qualitative and quantitative predictor of
a tested
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compound's in vivo pharmacology. See, for example, "Interaction of
Glucocorticoid
Analogues with the Human Glucocorticoid Receptor", T. S. Berger, Z.
Parandoosh, B. W.
Perry and R B. Stein, J. Steroid Biochem. Molec. Biol., 41 (1992) 733-738
(hereinafter
"Berger"), the disclosure of which is herein incorporated by reference.
In the cotransfection assay, a cloned cDNA for an IR (e.g., human PR, AR or
GR)
under the control of a constitutive promoter (e.g., the SV 40 promoter) is
introduced by
transfection (a procedure to induce cells to take up foreign genes) into a
background cell
substantially devoid of endogenous IRs. This introduced gene directs the
recipient cells to
make the IR protein of interest. A second gene is also introduced
(cotransfected) into the
same cells in conjunction with the IR gene. This second gene, comprising the
cDNA for a
reporter protein, such as firefly luciferase (LUC), is controlled by an
appropriate hormone
responsive promoter containing a hormone response element (HRE). This reporter
plasmid
functions as a reporter for the transcription-modulaxing activity of the
target IR. Thus, the
reporter acts as a surrogate for the products (mRNA, then protein) normally
expressed by a
gene under control of the target receptor and its native hormone.
The cotransfection assay can detect small molecule agonists or antagonists of
target
IRs. Exposing the transfected cells to an agonist ligand compound increases
reporter activity
in the transfected cells. This activity can be conveniently measured, e.g., by
increasing
luciferase production, which reflects compound-dependent, IR-mediated
increases in reporter
transcription. To detect antagonists, the cotransfection assay is carried out
in the presence of
a constant concentration of an agonist to the target IR (e.g., progesterone
for PR) known to
induce a defined reporter signal. Increasing concentrations of a suspected
antagonist will
decrease the reporter signal (e.g., luciferase production). The cotransfection
assay is
therefore useful to detect both agonists and antagonists of specific IRs.
Furthermore, it
determines not only whether a compound interacts with a particular IR, but
whether this
interaction mimics (agonizes) or blocks (antagonizes) the effects of the
native regulatory
molecules on target gene expression, as well as the specificity and strength
of this
interaction.
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The activities of selected steroid receptor modulator compounds of the present
invention were evaluated utilizing the cotransfection assay, and in standard
IR binding
assays, according to the following illustrative Examples.
EXAMPLE 6
Cotransfection assay
The fimction and detailed preparation procedure of the cotransfection assays
have
been described previously. See, for example, "Nonsteroidal Human Progesterone
Receptor
Modulators from the Marine Alga Cymopolia barbata", C. Pathirana, R. B. Stein,
T. S.
Bergen W. Fenical, T. Ianiro, D. E. Mais, A. Tomes, M. E. Goldman, Mol.
Pharm., 47
(1995) 630-635 (hereinafter "Pathirana"). Briefly, the cotransfection assays
were carried out
in CV-1 cells (African green monkey kidney fibroblasts) that had been
transiently transfected,
using the standard calcium phosphate coprecipitation procedure (see, e.g.,
Berger), with
plasmid containing receptor, MTV-LUC reporter, pRS-[3-Gal and filler DNA (Rous
sarcoma
virus chloramphenical acetyltransferase). The agonist activity was detemnined
by examining
the LUC expression (normalized response), and the efficacy readout was a
relative value to
the maximal LUC expression produced by a reference agonist, e.g., progesterone
for hPR,
dihydrotestosterone (DHT) for hAR, dexamethasone for hGR, aldosterone for hMR,
estradiol for hER. All the cotransfection experiments were carried out in 96-
well plates by
automation (Beckman Biomomek automated workstation).
Receutor Binding Assays
The preparation of receptor binding assays for hPR-A, hGR, and AR has been
described (see, e.g., Pathirana).
The agonist, antagonist and binding activity assay results of selected
progesterone
receptor modulator compounds of the present invention and the standard
reference
compounds on PR, as well as the cross-reactivity of selected compounds on the
AR, ER,
MR and GR receptors, are shown in Tables 1-2 below. Efficacy is reported as
the percent
maximal response observed for each compound relative to the reference agonist
and
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antagonist compounds indicated above. Also reported in Tables 1-2 for each
compound is
its antagonist potency or ICso (which is the concentration (nM), required to
reduce the
maximal response by 50%), its agonist potency or ECso (nM).
Table 1: Agonist, antagonist and binding activity of progesterone receptor
modulator
compounds of present invention and the reference agonist compound,
progesterone (Prog), and reference antagonists compound,RU486 and ZK299.
PR Agonist PR Antagonist PR
Cm d CV-1 Cells CV-1 Bindin
Cells
Efficacy Potency EfficacyPotency K;
No. (%) (nM) (%) (nM) (nM)
Prog 100 2.9 na na 3.5
RU486 na na 96 0.18 0.58
ZK299 na na 99 1.6 18
14 30 2500 95 25 172
20 30 500 80 20 17
24 na na 84 98 181
25 46 623 87 23 65
26 60 1000 68 16 ~ 20
na = not active (i.e. e~cacy of <20 and potency of >10,000)
nt = not tested
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Table 2: Overall agonist and antagonist potency of selected progesterone
receptor
modulator compounds of present invention and the reference
agonist and antagonist compounds shown in Table 1 on PR,
AR, ER, GR and MR.
GR MR
Cm d PR AR Potenc ER Potenc Potenc
Potenc Potenc
Agon Antag Agon Antag Agon Antag Antag Antag
No. (nM) (nM) (nM) (nM) (nM) (nM) (nM) (nM)
14 2500 25 na 200 na na na na
20 500 20 na 1000 na na na na
24 na 98 na 130 na na na na
25 623 23 na 100 na na na na
26 1000 16 na 300 na na na na
Pro 3 na 1300 na na na na nt
RU486 na 0.1 na 12 na 1500 0.7 1100
DHT na 1800 6 na 1700 na na nt
Flut na 1900 na 26 na na na na
Estr nt nt na na 7 na na nt
ICI164 na na na na na 160 na na
S it nt 268 nt nt na na 2000 25
na=not active (i.e., efficacy of >20 and potency of >10,000); nt=not tested
Pharmacological and Other Applications
As will be discernible to those skilled in the art, the PR modulator compounds
of the
present invention can be readily utilized in pharmacological applications
where PR antagonist
or agonist activity is desired, and where it is desired to minimize cross
reactivities with other
steroid receptor related IRs. In vivo applications of the invention include
administration of
the disclosed compounds to mammalian subjects, and in particular to humans.
The following Example provides illustrative pharmaceutical composition
formulations:
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EXAMPLE 7
Hard gelatin capsules are prepared using the following ingredients:
Quantity
(m~/cansule)
COMPOUND 26 140
Starch, dried 100
Magnesium stearate 10
Total (mg) 250
The above ingredients are mixed and filled into hard gelatin capsules in 250
mg quantities.
A tablet is prepared using the ingredients below:
Quantity
(mg/tablet)
COMPOUND 26 140
Cellulose, microcrystalline 200
Silicon dioxide, fumed 10
Stearic acid 10
Total (mg) 360
The components are blended and compressed to form tablets each weighing 360
mg.
Tablets, each containing 60 mg of active ingredient, are made as follows:
Quantity
(m~/tablet)
COMPOUND 26 60.0
Starch 45.0
Cellulose, microcrystalline 35.0
Polyvinylpyrrolidone (PVP)
(as 10% solution in water) 4.0
Sodium carboxymethyl starch (SCMS) 4.5
Magnesium stearate 0.5
Talc 1-00
Total (mg) 150.0
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The active ingredient, starch, and cellulose are passed through a No. 45 mesh
U. S. sieve and
mixed thoroughly. The solution of PVP is mixed with the resultant powders,
which are then
passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at
50 °C and
passed through a No. 18 mesh U. S. sieve. The SCMS, magnesium stearate, and
talc,
previously passed through a No. 60 mesh U.S. sieve, and then added to the
granules which,
after mixing, are compressed on a tablet machine to yield tablets each
weighing 150 mg.
Suppositories, each containing 225 mg of active ingredient, may be made as
follows:
COMPOUND 26 225 mg
Saturated fatty acid glycerides 2,000 ma
Total 2,225 mg
The active ingredient is passed through a No. 60 mesh U. S. sieve and
suspended in the
saturated fatty acid glycerides previously melted using the minimum heat
necessary. The
mixture is then poured into a suppository mold of normal 2g capacity and
allowed to cool.
An intravenous formulation may be prepared as follows:
COMPOUND 26 100 mg
Isotonic saline 1,000 mL
Glycerol 100 mL
The compound is dissolved in the glycerol and then the solution is slowly
diluted with
isotonic saline. The solution of the above ingredients is then administered
intravenously at a
rate of 1 mL per minute to a patient.
While in accordance with the patent statutes, description of the preferred
embodiments and processing conditions have been provided, the scope of the
invention is not
to be limited thereto or thereby. Various modifications and alterations of the
present
invention will be apparent to those skilled in the art without departing from
the scope and
spirit of the present invention.
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Consequently, for an understanding of the scope of the present invention,
reference is
made to the following claims.
