U.S. patent application number 10/050287 was filed with the patent office on 2002-08-15 for cyclic urea and cyclic amide derivatives.
This patent application is currently assigned to American Home Products Corporation. Invention is credited to Edwards, James P., Fensome, Andrew, Jones, Todd K., Santilli, Arthur A., Tegley, Christopher M., Terefenko, Eugene A., Viet, Andrew Q., Wrobel, Jay E., Zhang, Puwen, Zhi, Lin.
Application Number | 20020111355 10/050287 |
Document ID | / |
Family ID | 56290008 |
Filed Date | 2002-08-15 |
United States Patent
Application |
20020111355 |
Kind Code |
A1 |
Zhang, Puwen ; et
al. |
August 15, 2002 |
Cyclic urea and cyclic amide derivatives
Abstract
This invention provides compounds of the formula: 1 wherein: A,
B and D are N or CH, with the proviso that A, B and D cannot all be
CH; R.sup.1 and R.sup.2 are independent substituents selected from
H, COR.sup.A, NR.sup.BCOR.sup.A, or as defined herein; or R.sup.1
and R.sup.2 are fused to form an optionally substituted 3 to 8
membered spirocyclic or heterocyclic ring; R.sup.3 is H, OH,
NH.sub.2, or optionally substituted alkyl, or alkenyl, or
COR.sup.C; R.sup.4 is a substituted benzene ring or a five or six
membered ring with 1, 2, or 3 heteroatoms from the group defined
herein or Q is O, S, NR.sup.6, or CR.sup.7R.sup.8; R.sup.6, R.sup.7
and R.sup.8 are as defined herein; or CR.sup.7R.sup.8 form a six
membered ring; W is O or a chemical bond; as well as their use and
pharmaceutical compositions as agonists and antagonists of the
progesterone receptor.
Inventors: |
Zhang, Puwen; (Audubon,
PA) ; Santilli, Arthur A.; (Havertown, PA) ;
Fensome, Andrew; (Wayne, PA) ; Terefenko, Eugene
A.; (Quakertown, PA) ; Viet, Andrew Q.; (Upper
Darby, PA) ; Wrobel, Jay E.; (Lawrenceville, NJ)
; Edwards, James P.; (San Diego, CA) ; Jones, Todd
K.; (Solana Beach, CA) ; Tegley, Christopher M.;
(Thousand Oaks, CA) ; Zhi, Lin; (San Diego,
CA) |
Correspondence
Address: |
HOWSON AND HOWSON
ONE SPRING HOUSE CORPORATION CENTER
BOX 457
321 NORRISTOWN ROAD
SPRING HOUSE
PA
19477
US
|
Assignee: |
American Home Products
Corporation
Madison
NJ
07840
|
Family ID: |
56290008 |
Appl. No.: |
10/050287 |
Filed: |
January 16, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10050287 |
Jan 16, 2002 |
|
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09552356 |
Apr 19, 2000 |
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60183037 |
May 4, 1999 |
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Current U.S.
Class: |
514/243 ;
514/248; 514/251; 514/262.1; 544/184; 544/231; 544/236; 544/256;
544/257 |
Current CPC
Class: |
A61P 13/08 20180101;
A61P 1/00 20180101; A61P 5/36 20180101; A61P 35/00 20180101; C07D
498/04 20130101; A61P 15/00 20180101; A61P 15/18 20180101; A61P
5/24 20180101; A61P 5/00 20180101; A61P 15/16 20180101; A61P 15/08
20180101; A61P 5/34 20180101 |
Class at
Publication: |
514/243 ;
514/248; 514/251; 544/184; 544/231; 544/236; 514/262.1; 544/256;
544/257 |
International
Class: |
A61K 031/53; A61K
031/525; A61K 031/519; C07D 487/02 |
Claims
What is claimed:
1. A compound of the formula: 34wherein: A, B and D are N or CH,
with the proviso that two or three of A, B and D are N; R.sup.1 and
R.sup.2 are independent substituents selected from the group
consisting of H, C.sub.1 to C.sub.6 alkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, COR.sup.A, and
NR.sup.BCOR.sup.A; or R.sup.1 and R.sup.2 are fused to form a
spirocyclic ring selected from the group consisting of a), b) and
c): a) a 3 to 8 membered saturated spirocyclic ring; b) a 3 to 8
membered spirocyclic ring having one or more carbon-carbon double
bonds; and c) a 3 to 8 membered heterocyclic ring having in its
backbone one to three heteroatoms selected from the group
consisting of O, S and N; the spirocyclic rings of a), b) and c)
being optionally substituted by from 1 to 4 groups selected from
the group consisting of fluorine, C.sub.1 to C.sub.6 alkyl, C.sub.1
to C.sub.6 alkoxy, C.sub.1 to C.sub.6 thioalkyl, CF.sub.3, OH, CN,
NH.sub.2, NH(C.sub.1 to C.sub.6 alkyl), and N(C.sub.1 to C.sub.6
alkyl).sub.2; R.sup.A is H, C.sub.1 to C.sub.3 alkyl, substituted
C.sub.1 to C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to
C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to
C.sub.3 aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl;
R.sup.B is H, C.sub.1 to C.sub.3 alkyl, or substituted C.sub.1 to
C.sub.3 alkyl; R.sup.3 is H, OH, NH.sub.2, C.sub.1 to C.sub.6
alkyl, substituted C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.6
alkenyl, substituted C.sub.3 to C.sub.6 alkenyl, or COR.sup.C;
R.sup.C is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to
C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy,
substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.4
is (i) or (i): (i) a substituted benzene ring having the
substituents X, Y and Z as shown below: 35wherein: X is selected
from the group consisting of halogen, CN, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3 alkoxy,
substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
thioalkoxy, substituted C.sub.1 to C.sub.3 thioalkoxy, amino,
C.sub.1 to C.sub.3 aminoalkyl, substituted C.sub.1 to C.sub.3
aminoalkyl, NO.sub.2, C.sub.1 to C.sub.3 perfluoroalkyl, 5 or 6
membered heterocyclic ring having in its backbone 1 to 3
heteroatoms, COR.sup.D, OCOR.sup.D, and NR.sup.ECOR.sup.D; R.sup.D
is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3
alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy,
substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.E
is H, C.sub.1 to C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3
alkyl; Y and Z are independent substituents selected from the group
consisting of H, halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 alkyl, and C.sub.1 to C.sub.3 thioalkoxy; or
(ii) a five or six membered ring having in its backbone 1, 2, or 3
heteroatoms selected from the group consisting of O, S, SO,
SO.sub.2 and NW and having one or two independent substituents
selected from the group consisting of H, halogen, CN, NO.sub.2,
C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3 alkoxy, C.sub.1 to
C.sub.3 aminoalkyl, COR.sup.F, and NR.sup.GCOR.sup.F; R.sup.F is H,
C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy, substituted
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or
substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.G is H, C.sub.1 to
C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3 alkyl; R.sup.5 is
H or C.sub.1 to C.sub.3 alkyl; Q is O or S; or a pharmaceutically
acceptable salt thereof.
2. The compound according to claim 1, wherein: R.sup.1 and R.sup.2
are H, C.sub.1 to C.sub.6 alkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, COR.sup.A, or
NR.sup.BCOR.sup.A; R.sup.3is H, OH, NH.sub.2, C.sub.1 to C.sub.6
alkyl, substituted C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.6
alkenyl, substituted C.sub.3 to C.sub.6 alkenyl, or COR.sup.C;
R.sup.4is (iii) or (iv): (iii) the substituted benzene ring,
wherein: X is selected from the group consisting of halogen, CN,
C.sub.1 to C.sub.3alkyl, substituted C.sub.1 to C.sub.3 alkyl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 thioalkoxy, substituted C.sub.1 to C.sub.3
thioalkoxy, C.sub.1 to C.sub.3 aminoalkyl, substituted C.sub.1 to
C.sub.3 aminoalkyl, NO.sub.2, C.sub.1 to C.sub.3 perfluoroalkyl, a
5 membered heterocyclic ring having in its backbone 1 to 3
heteroatoms, COR.sup.D, OCOR.sup.D, and NR.sup.ECOR.sup.D; or (iv)
the five or six membered ring having one or two independent
substituents selected from the group consisting of H, halogen, CN,
NO.sub.2, C.sub.1 to C.sub.3 alkyl, and C.sub.1 to C.sub.3
alkoxy.
3. The compound according to claim 1, wherein: R.sup.1=R.sup.2 and
are C.sub.1 to C.sub.3 alkyl or R.sup.1 and R.sup.2 are fused to
form the 3 to 6 membered saturated spirocyclic ring; R.sup.3 is H,
OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, or COR.sup.C; R.sup.C is H, C.sub.1 to C.sub.3
alkyl, or C.sub.1 to C.sub.3 alkoxy; R.sup.4 is selected from the
group consisting of (v), (vi), and (vii): (v) the substituted
benzene ring of the formula: 36wherein: X is selected from the
group consisting of of halogen, CN, C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 alkyl, NO.sub.2, C.sub.1 to C.sub.3
perfluoroalkyl, 5 membered heterocyclic ring having in its backbone
1 to 3 heteroatoms, and C.sub.1 to C.sub.3 thioalkoxy; Y is on the
4' or 5' position and is selected from the group consisting of H,
halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkoxy, C.sub.1 to
C.sub.3 alkyl, and C.sub.1 to C.sub.3 thioalkoxy; (vi) the five
membered ring of the structure: 37wherein: U is O, S, or NR.sup.5;
X' is selected from the group consisting of halogen, CN, NO.sub.2,
C.sub.1 to C.sub.3 alkyl, and C.sub.1 to C.sub.3 alkoxy; Y' is
selected from the group consisting of H and C.sub.1 to C.sub.3
alkyl; and (vii) the six membered ring of the structure: 38wherein:
X.sup.1 is N or CX.sup.2; X.sup.2 is halogen, CN or NO.sub.2.
4. The compound according to claim 1, wherein: R.sup.1=R.sup.2 and
are CH.sub.3 or R.sup.1 and R.sup.2 are fused to form a 6 membered
saturated spirocyclic ring; R.sup.3 is H, OH, NH.sub.2, CH.sub.3,
substituted CH.sub.3, or COR.sup.C; R.sup.C is H, C.sub.1 to
C.sub.3 alkyl, or C.sub.1 to C.sub.3 alkoxy; R.sup.4 is the
substituted benzene ring having the substituents X and Y as shown
below: 39wherein: X is halogen, CN, C.sub.1 alkoxy, NO.sub.2, or
5-membered heterocyclic ring; wherein said heterocyclic ring is
2-thiazole; Y is on the 4' or 5' position and is H or halogen;
wherein said halogen is F.
5. The compound according to claim 4 wherein R.sup.4 is the moiety:
40
6. The compound according to claim 1, wherein: R.sup.1=R.sup.2 and
are CH.sub.3 or R.sup.1 and R.sup.2 are fused to form a 6 membered
saturated spirocyclic ring; R.sup.3 is H, OH, NH.sub.2, CH.sub.3,
substituted CH.sub.3, or COR.sup.C; R.sup.C is H, C.sub.1 to
C.sub.3 alkyl, or C.sub.1 to C.sub.3 alkoxy; R.sup.4 is the five
membered ring of the structure: 41U is O, S, or NH; X' is halogen,
CN, or NO.sub.2; Y' is H or C.sub.1 to C.sub.3 alkyl.
7. The compound according to claim 6 wherein R.sup.4 is the moiety:
42wherein: Y' is H or C.sub.1 to C.sub.3 alkyl.
8. A pharmaceutical composition comprising a pharmaceutically
effective amount of a compound of claim 1 and a pharmaceutically
effective carrier or excipient.
9. A method of inducing contraception in a mammal, the method
comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound of claim 1 or a
pharmaceutically acceptable salt thereof.
10. A method of treatment or prevention of benign or malignant
neoplastic disease in a mammal, the method comprising administering
to a mammal in need thereof a compound of claim 1, or a
pharmaceutically acceptable salt thereof.
11. The method according to claim 10, wherein the benign or
malignant neoplastic disease is selected from the group consisting
of uterine myometrial fibroids, endometriosis, benign prostatic
hypertrophy; carcinomas or adenocarcinomas of the endometrium,
ovary, breast, colon, prostate, pituitary, meningioma and other
hormone-dependent tumors.
12. A method of treatment or prevention in a mammal of carcinomas
or adenocarcinomas of the endometrium, ovary, breast, colon, or
prostate, the method comprising administering to a mammal in need
thereof a compound of claim 1, or a pharmaceutically acceptable
salt thereof.
13. A compound of the formula: 43wherein: A, B and D are N or CH,
with the proviso that two or more of A, B and D is N; R.sup.1 and
R.sup.2 are independent substituents selected from the group
consisting of H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.2 to C.sub.6 alkenyl, substituted C.sub.2 to
C.sub.6 alkenyl, C.sub.2 to C.sub.6 alkynyl, substituted C.sub.2 to
C.sub.6 alkynyl, C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3
to C.sub.8 cycloalkyl, aryl, substituted aryl, heterocyclic,
substituted heterocyclic, COR.sup.A, and NR.sup.BCOR.sup.A; or
R.sup.1 and R.sup.2 are fused to form a spirocyclic ring selected
from the group consisting of a), b) and c): a) a 3 to 8 membered
saturated spirocyclic ring; b) a 3 to 8 membered spirocyclic ring
having one or more carbon-carbon double bonds; and c) a 3 to 8
membered heterocyclic ring having in its backbone one to three
heteroatoms selected from the group consisting of O, S and N; the
spirocyclic rings of a), b) and c) being optionally substituted by
from 1 to 4 groups selected from the group consisting of fluorine,
C.sub.1 to C.sub.6 alkyl, C.sub.1 to C.sub.6 alkoxy, C.sub.1 to
C.sub.6 thioalkyl, CF.sub.3, OH, CN, NH.sub.2, NH(C.sub.1 to
C.sub.6 alkyl), and N(C.sub.1 to C.sub.6 alkyl).sub.2; R.sup.A is
H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy, substituted
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or
substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.B is H, C.sub.1 to
C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3 alkyl; R.sup.3 is
H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.3 to C.sub.6 alkenyl, substituted C.sub.3 to
C.sub.6 alkenyl, or COR.sup.C; R.sup.C is H, C.sub.1 to C.sub.3
alkyl, substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted
aryl, C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to
C.sub.3 aminoalkyl; R.sup.4 is (i) or (ii): (i) a substituted
benzene ring having the substituents X, Y and Z as shown below:
44wherein: X is selected from the group consisting of halogen, CN,
C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 thioalkoxy, substituted C.sub.1 to C.sub.3
thioalkoxy, amino, C.sub.1 to C.sub.3 aminoalkyl, substituted
C.sub.1 to C.sub.3 amino alkyl, NO.sub.2, C.sub.1 to C.sub.3
perfluoroalkyl, 5 or 6 membered heterocyclic ring having in its
backbone 1 to 3 heteroatoms, COR.sup.D, OCOR.sup.D, and
NR.sup.ECOR.sup.D; R.sup.D is H, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted aryl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to C.sub.3
aminoalkyl; R.sup.E is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl; Y and Z are independent substituents
selected from the group consisting of H, halogen, CN, NO.sub.2,
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 alkyl, and C.sub.1 to
C.sub.3 thioalkoxy; or (ii) a five or six membered ring having in
its backbone 1, 2, or 3 heteroatoms selected from the group
consisting of O, S, SO, SO.sub.2 and NR.sup.5 and having one or two
independent substituents selected from the group consisting of H,
halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, COR.sup.F, and
NR.sup.GCOR.sup.F; R.sup.F is H, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted aryl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to C.sub.3
aminoalkyl; R.sup.G is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl; R.sup.5 is H or C.sub.1 to C.sub.3 alkyl;
Q is O, S, NR.sup.6, or CR.sup.7R; R.sub.6 is selected from the
group consisting of CN, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl,
substituted C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, and SO.sub.2CF.sub.3;
R.sup.7 and R.sup.8 are independent substituents selected from the
group consisting of H, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl,
substituted C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, NO.sub.2, CN, and
CO.sub.2R.sup.9; R.sup.9 is C.sub.1 to C.sub.3 alkyl; or
CR.sup.7R.sup.8 form a six membered ring having the structure
below: 45W is a chemical bond; or a pharmaceutically acceptable
salt thereof.
14. A compound of the formula: 46wherein: A, B and D are N or CH,
with the proviso that two or more of A, B and D is N; R.sup.1 and
R.sup.2 are independent substituents selected from the group
consisting of H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.2 to C.sub.6 alkenyl, substituted C.sub.2 to
C.sub.6 alkenyl, C.sub.2 to C.sub.6 alkynyl, substituted C.sub.2 to
C.sub.6 alkynyl, C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3
to C.sub.8 cycloalkyl, aryl, substituted aryl, heterocyclic,
substituted heterocyclic, COR.sup.A, and NR.sup.ECOR.sup.A; or
R.sup.1 and R.sup.2 are fused to form a spirocyclic ring selected
from the group consisting of a), b) and c): a) a 3 to 8 membered
saturated spirocyclic ring; b) a 3 to 8 membered spirocyclic ring
having one or more carbon-carbon double bonds; and c) a 3 to 8
membered heterocyclic ring having in its backbone one to three
heteroatoms selected from the group consisting of O, S and N; the
spirocyclic rings of a), b) and c) being optionally substituted by
from 1 to 4 groups selected from the group consisting of fluorine,
C.sub.1 to C.sub.6 alkyl, C.sub.1 to C.sub.6 alkoxy, C.sub.1 to
C.sub.6 thioalkyl, CF.sub.3, OH, CN, NH.sub.2, NH(C.sub.1 to
C.sub.6 alkyl), and N(C.sub.1 to C.sub.6 alkyl).sub.2; R.sup.A is
H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy, substituted
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or
substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.B is H, C.sub.1 to
C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3 alkyl; R.sup.3 is
H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.3 to C.sub.6 alkenyl, substituted C.sub.3 to
C.sub.6 alkenyl, or COR.sup.C; R.sup.C is H, C.sub.1 to C.sub.3
alkyl, substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted
aryl, C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to
C.sub.3 aminoalkyl; R.sup.4 is (i) or (ii): (i) a substituted
benzene ring having the substituents X, Y and Z as shown below:
47wherein: X is selected from the group consisting of halogen, CN,
C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 thioalkoxy, substituted C.sub.1 to C.sub.3
thioalkoxy, amino, C.sub.1 to C.sub.3 aminoalkyl, substituted
C.sub.1 to C.sub.3 aminoalkyl, NO.sub.2, C.sub.1 to C.sub.3
perfluoroalkyl, 5 or 6 membered heterocyclic ring having in its
backbone 1 to 3 heteroatoms, COR.sup.D, OCOR.sup.D, and
NR.sup.BCOR.sup.D; R.sup.D is H, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted alkyl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 amino alkyl, or substituted C.sub.1 to C.sub.3
aminoalkyl; R.sup.E is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl; Y and Z are independent substituents
selected from the group consisting of H, halogen, CN, NO.sub.2,
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 alkyl, and C.sub.1 to
C.sub.3 thioalkoxy; or (ii) a five or six membered ring having in
its backbone 1, 2, or 3 heteroatoms selected from the group
consisting of O, S, SO, SO.sub.2 and NR.sup.5 and having one or two
independent substituents selected from the group consisting of H,
halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, COR.sup.F, and
NR.sup.GCOR.sup.F; R.sup.F is H, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted aryl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to C.sub.3
aminoalkyl; R.sup.G is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl; R.sup.5is H or C.sub.1 to C.sub.3 alkyl;
Q is NR.sup.6 or CR.sup.7R.sup.8; R.sub.6 is selected from the
group consisting of CN, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl,
substituted C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, and SO.sub.2CF.sub.3;
R.sup.7 and R.sup.8 are independent substituents selected from the
group consisting of H, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to Cg cycloalkyl, substituted
C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, NO.sub.2, CN, and
CO.sub.2R.sup.9; R.sup.9 is C.sub.1 to C.sub.3 alkyl; or
CR.sup.7R.sup.8 form a six membered ring having the structure
below: 48W is O; or a pharmaceutically acceptable salt thereof.
15. A compound of the formula: 49wherein: A, B and D are N or CH,
with the proviso that two or three of A, B and D are N; R.sup.1 and
R.sup.2 are independent substituents selected from the group
consisting of H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.2 to C.sub.6 alkenyl, substituted C.sub.2 to
C.sub.6 alkenyl, C.sub.2 to C.sub.6 alkynyl, substituted C.sub.2 to
C.sub.6 alkynyl, C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3
to C.sub.8 cycloalkyl, aryl, substituted aryl, heterocyclic,
substituted heterocyclic, COR.sup.A, and NR.sup.BCOR.sup.A; or
R.sup.1 and R.sup.2 are fused to form a spirocyclic ring selected
from the group consisting of a), b) and c): a) a 3 to 8 membered
saturated spirocyclic ring; b) a 3 to 8 membered spirocyclic ring
having one or more carbon-carbon double bonds; and c) a 3 to 8
membered heterocyclic ring having in its backbone one to three
heteroatoms selected from the group consisting of O, S and N; the
spirocyclic rings of a), b) and c) being optionally substituted by
from 1 to 4 groups selected from the group consisting of fluorine,
C.sub.1 to C.sub.6 alkyl, C.sub.1 to C.sub.6 alkoxy, C.sub.1 to
C.sub.6 thioalkyl, CF.sub.3, OH, CN, NH.sub.2, NH(C.sub.1 to
C.sub.6 alkyl), and N(C.sub.1 to C.sub.6 alkyl).sub.2; R.sup.A is
H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy, substituted
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or
substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.B is H, C.sub.1 to
C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3 alkyl; R.sup.3 is
H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.3 to C.sub.6 alkenyl, substituted C.sub.3 to
C.sub.6 alkenyl, or COR.sup.C; R.sup.C is H, C.sub.1 to C.sub.3
alkyl, substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted
aryl, C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to
C.sub.3 aminoalkyl; R.sup.4 is a substituted benzene ring having
the substituents X, Y and Z as shown below: 50wherein: X is
selected from the group consisting of halogen, CN, C.sub.1 to
C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl, C.sub.1 to
C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to
C.sub.3 thioalkoxy, substituted C.sub.1 to C.sub.3 thioalkoxy,
amino, C.sub.1 to C.sub.3 aminoalkyl, substituted C.sub.1 to
C.sub.3 aminoalkyl, NO.sub.2, C.sub.1 to C.sub.3 perfluoroalkyl, 5
or 6 membered heterocyclic ring having in its backbone 1 to 3
heteroatoms, COR.sup.D, OCOR.sup.D, and NR.sup.ECOR.sup.D; R.sup.D
is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3
alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy,
substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 amino alkyl; R.sup.E
is H, C.sub.1 to C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3
alkyl; Y and Z are independent substituents selected from the group
consisting of halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 alkyl, and C.sub.1 to C.sub.3 thioalkoxy; Q is
O, S, NW, or CR.sup.7R.sup.8; R.sub.6 is selected from the group
consisting of CN, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3
to C.sub.8 cycloalkyl, aryl, substituted aryl, heterocyclic,
substituted heterocyclic, and SO.sub.2CF.sub.3; R.sup.7 and R.sup.8
are independent substituents selected from the group consisting of
H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to C.sub.6 alkyl,
C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3 to C.sub.8
cycloalkyl, aryl, substituted aryl, heterocyclic, substituted
heterocyclic, NO.sub.2, CN, and CO.sub.2R.sup.9; R.sup.9 is C.sub.1
to C.sub.3 alkyl; or CR.sup.7R.sup.8 form a six membered ring
having the structure below: 51W is O; or a pharmaceutically
acceptable salt thereof.
16. A compound of the formula: 52wherein: A, B and D are N or CH,
with the proviso that one of A, B and D is N; R.sup.1 and R.sup.2
are independent substituents selected from the group consisting of
H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to C.sub.6 alkyl,
C.sub.2 to C.sub.6 alkenyl, substituted C.sub.2 to C.sub.6 alkenyl,
C.sub.2 to C.sub.6 alkynyl, substituted C.sub.2 to C.sub.6 alkynyl,
C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3 to C.sub.8
cycloalkyl, aryl, substituted aryl, heterocyclic, substituted
heterocyclic, COR.sup.A, and NR.sup.BCOR.sup.A; or R.sup.1 and
R.sup.2 are fused to form a spirocyclic ring selected from the
group consisting of a), b) and c): a) a 3 to 8 membered saturated
spirocyclic ring; b) a 3 to 8 membered spirocyclic ring having one
or more carbon-carbon double bonds; and c) a 3 to 8 membered
heterocyclic ring having in its backbone one to three heteroatoms
selected from the group consisting of O, S and N; the spirocyclic
rings of a), b) and c) being optionally substituted by from 1 to 4
groups selected from the group consisting of fluorine, C.sub.1 to
C.sub.6 alkyl, C.sub.1 to C.sub.6 alkoxy, C.sub.1 to C.sub.6
thioalkyl, CF.sub.3, OH, CN, NH.sub.2, NH(C.sub.1 to C.sub.6
alkyl), and N(C.sub.1 to C.sub.6 alkyl).sub.2; R.sup.A is H,
C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy, substituted
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or
substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.B is H, C.sub.1 to
C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3 alkyl; R.sup.3 is
H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.3 to C.sub.6 alkenyl, substituted C.sub.3 to
C.sub.6 alkenyl, or COR.sup.C; R.sup.C is H, C.sub.1 to C.sub.3
alkyl, substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted
aryl, C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to
C.sub.3 aminoalkyl; R.sup.4 is (i), (ii), (iii), or (iv): (i) a
substituted benzene ring having the substituents X, Y and Z as
shown below: 53wherein: X is selected from the group consisting of
halogen, CN, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to
C.sub.3 alkyl, C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to
C.sub.3 alkoxy, C.sub.1 to C.sub.3 thioalkoxy, substituted C.sub.1
to C.sub.3 thioalkoxy, amino, C.sub.1 to C.sub.3 aminoalkyl,
substituted C.sub.1 to C.sub.3 aminoalkyl, NO.sub.2, C.sub.1 to
C.sub.3 perfluoroalkyl, 5 or 6 membered heterocyclic ring having in
its backbone 1 to 3 heteroatoms, COR.sup.D, OCOR.sup.D, and
NR.sup.ECOR.sup.D; R.sup.D is H, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted aryl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to C.sub.3
aminoalkyl; R.sup.E is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl; Y and Z are independent substituents
selected from the group consisting of H, halogen, CN, NO.sub.2,
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 alkyl, and C.sub.1 to
C.sub.3 thioalkoxy; (ii) a five membered ring having in its
backbone 1, 2, or 3 heteroatoms selected from the group consisting
of O, S, SO, SO.sub.2 and NR.sup.5 and having one or two
independent substituents selected from the group consisting of H,
halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, COR.sup.F, and
NR.sup.GCOR.sup.F; (iii) a six membered ring having in its backbone
1 NR.sup.5 heteroatom and having one or two independent
substituents selected from the group consisting of H, halogen, CN,
NO.sub.2, C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 aminoalkyl, COR.sup.F, and NR.sup.GCOR.sup.F; or
(iv) a six membered ring having in its backbone 2 or 3 heteroatoms
selected from the group consisting of O, S, SO, and SO.sub.2 and
having one or two independent substituents selected from the group
consisting of H, halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkyl,
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl,
COR.sup.F, and NR.sup.GCOR.sup.F; R.sup.F is H, C.sub.1 to C.sub.3
alkyl, substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted
aryl, C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to
C.sub.3 aminoalkyl; R.sup.G is H, C.sub.1 to C.sub.3 alkyl, or
substituted C.sub.1 to C.sub.3 alkyl; R.sup.5 is H or C.sub.1 to
C.sub.3 alkyl; Q is O; W is a chemical bond; or a pharmaceutically
acceptable salt thereof.
17. A compound of the formula: 54wherein: A, B and D are N or CH,
with the proviso that one of A, B and D is N; R.sup.1 and R.sup.2
are independent substituents selected from the group consisting of
H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to C.sub.6 alkyl,
C.sub.2 to C.sub.6 alkenyl, substituted C.sub.2 to C.sub.6 alkenyl,
C.sub.2 to C.sub.6 alkynyl, substituted C.sub.2 to C.sub.6 alkynyl,
C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3 to C.sub.8
cycloalkyl, aryl, substituted aryl, heterocyclic, substituted
heterocyclic, COR.sup.A, and NR.sup.BCOR.sup.A; or R.sup.1 and
R.sup.2 are fused to form a spirocyclic ring selected from the
group consisting of a), b) and c): a) a 3 to 8 membered saturated
spirocyclic ring; b) a 3 to 8 membered spirocyclic ring having one
or more carbon-carbon double bonds; and c) a 3 to 8 membered
heterocyclic ring having in its backbone one to three heteroatoms
selected from the group consisting of O, S and N; the spirocyclic
rings of a), b) and c) being optionally substituted by from 1 to 4
groups selected from the group consisting of fluorine, C.sub.1 to
C.sub.6 alkyl, C.sub.1 to C.sub.6 alkoxy, C.sub.1 to C.sub.6
thioalkyl, CF.sub.3, OH, CN, NH.sub.2, NH(C.sub.1 to C.sub.6
alkyl), and N(C.sub.1 to C.sub.6 alkyl).sub.2; R.sup.A is H,
C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
aryl, substituted aryl, C.sub.1 to C.sub.3 alkoxy, substituted
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or
substituted C.sub.1 to C.sub.3 aminoalkyl; R.sup.B is H, C.sub.1 to
C.sub.3 alkyl, or substituted C.sub.1 to C.sub.3 alkyl; R.sup.3 is
H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.3 to C.sub.6 alkenyl, substituted C.sub.3 to
C.sub.6 alkenyl, or COR.sup.C; R.sup.C is H, C.sub.1 to C.sub.3
alkyl, substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted
aryl, C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to
C.sub.3 aminoalkyl; R.sup.4 is (i) or (ii): (i) a substituted
benzene ring having the substituents X, Y and Z as shown below:
55wherein: X is selected from the group consisting of halogen, CN,
C.sub.1 to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 thioalkoxy, substituted C.sub.1 to C.sub.3
thioalkoxy, amino, C.sub.1 to C.sub.3 aminoalkyl, substituted
C.sub.1 to C.sub.3 aminoalkyl, NO.sub.2, C.sub.1 to C.sub.3
perfluoroalkyl, 5 or 6 membered heterocyclic ring having in its
backbone 1 to 3 heteroatoms, COR.sup.D, OCOR.sup.D, and
NR.sup.ECOR.sup.D; R.sup.D is H, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted aryl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to C.sub.3
aminoalkyl; R.sup.E is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl; Y and Z are independent substituents
selected from the group consisting of H, halogen, CN, NO.sub.2,
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 alkyl, and C.sub.1 to
C.sub.3 thioalkoxy; or (ii) a five or six membered ring having in
its backbone 1, 2, or 3 heteroatoms selected from the group
consisting of O, S, SO, SO.sub.2 and NR.sup.5 and having one or two
independent substituents selected from the group consisting of H,
halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3
alkoxy, C.sub.1 to C.sub.3 aminoalkyl, COR.sup.F, and
NR.sup.ECOR.sup.F; R.sup.F is H, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, aryl, substituted aryl,
C.sub.1 to C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 aminoalkyl, or substituted C.sub.1 to C.sub.3
aminoalkyl; R.sup.G is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl; R.sup.5 is H or C.sub.1 to C.sub.3 alkyl;
Q is S, NR.sup.6, or CR.sup.7R.sup.8; R.sub.6 is selected from the
group consisting of CN, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl,
substituted C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, and SO.sub.2CF.sub.3;
R.sup.7 and R.sup.8 are independent substituents selected from the
group consisting of H, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl,
substituted C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, NO.sub.2, CN, and
CO.sub.2R.sup.9; R.sup.9 is C.sub.1 to C.sub.3 alkyl; or
CR.sup.7R.sup.8 form a six membered ring having the structure
below: 56W is a chemical bond; or a pharmaceutically acceptable
salt thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a divisional of U.S. patent application
Ser. No. 09/552,356, filed Apr. 19, 2000, which claims the benefit
of the priority of U.S. Patent Application No. 60/183,037, filed
May 4, 1999, now abandoned.
BACKGROUND OF THE INVENTION
[0002] This invention relates to compounds which are agonists and
antagonists of the progesterone receptor, their preparation and
utility.
[0003] Intracellular receptors (IR) form a class of structurally
related gene regulators known as "ligand dependent transcription
factors" (R. M. Evans, Science, 240, 889, 1988). The steroid
receptor family is a subset of the IR family, including
progesterone receptor (PR), estrogen receptor (ER), androgen
receptor (AR), glucocorticoid receptor (GR), and mineralocorticoid
receptor (MR).
[0004] The natural hormone, or ligand, for the PR is the steroid
progesterone, but synthetic compounds, such as medroxyprogesterone
acetate or levonorgestrel, have been made which also serve as
ligands. Once a ligand is present in the fluid surrounding a cell,
it passes through the membrane via passive diffusion, and binds to
the IR to create a receptor/ligand complex. This complex binds to
specific gene promoters present in the cell's DNA. Once bound to
the DNA the complex modulates the production of mRNA and protein
encoded by that gene.
[0005] A compound that binds to an IR and mimics the action of the
natural hormone is termed an agonist, whilst a compound that
inhibits the effect of the hormone is an antagonist.
[0006] PR agonists (natural and synthetic) are known to play an
important role in the health of women. PR agonists are used in
birth control formulations, typically in the presence of an ER
agonist. ER agonists are used to treat the symptoms of menopause,
but have been associated with a proliferative effect on the uterus
that can lead to an increased risk of uterine cancers.
Co-administration of a PR agonist reduces or ablates that risk.
[0007] PR antagonists may also be used in contraception. In this
context they may be administered alone (Ulmann, et al, Ann. N.Y
Acad. Sci., 261, 248, 1995), in combination with a PR agonist
(Kekkonen, et al, Fertility and Sterility, 60, 610, 1993) or in
combination with a partial ER antagonist such as tamoxifen (WO
96/19997 A1, Jul. 4, 1996).
[0008] PR antagonists may also be useful for the treatment of
hormone dependent breast cancers (Horwitz, et al, Horm Cancer, 283,
pub: Birkhaeuser, Boston, Mass., ed. Vedeckis) as well as uterine
and ovarian cancers. PR antagonists may also be useful for the
treatment of non-malignant chronic conditions such as fibroids
(Murphy, et al, J. Clin. Endo. Metab., 76, 513, 1993) and
endometriosis (Kettel, et al, Fertility and Sterility, 56, 402,
1991).
[0009] PR antagonists may also be useful in hormone replacement
therapy for post menopausal patients in combination with a partial
ER antagonist such as tamoxifen (U.S. Pat. No. 5,719,136).
[0010] PR antagonists, such as nifepristone and onapristone, have
been shown to be effective in a model of hormone dependent prostate
cancer, which may indicate their utility in the treatment of this
condition in men (Michna, et al, Ann. N. Y Acad. Sci., 761, 224,
1995).
[0011] Jones, et al, (U.S. Pat. No. 5,688,810) describe the PR
antagonist dihydroquinoline 1. 2
[0012] Jones, et al, described the enol ether 2 (U.S. Pat. No.
5,693,646) as a PR ligand. 3
[0013] Jones, et al, described compound 3 (U.S. Pat. No. 5,696,127)
as a PR ligand. 4
[0014] Zhi, et al, described lactones 4, 5 and 6 as PR antagonists
(J. Med. Chem, 41, 291, 1998). 5
[0015] Zhi, et al, described the ether 7 as a PR antagonist (J.
Med. Chem, 41, 291, 1998). 6
[0016] Combs, et al., disclosed the amide 8 as a ligand for the PR
(J. Med. Chem., 38, 4880, 1995). 7
[0017] Perlman, et al., described the vitamin D analog 9 as a PR
ligand (Tet. Letters, 35, 2295, 1994). 8
[0018] Hamann, et al, described the PR antagonist 10 (Ann. N. Y
Acad. Sci., 761, 383, 1995). 9
[0019] Chen, et al, described the PR antagonist 11 (Chen, et al,
POI-37, 16.sup.th Int. Cong. Het. Chem., Montana, 1997). 10
[0020] Kurihari, et. al., described the PR ligand 12 (J.
Antibiotics, 50, 360, 1997). 11
[0021] The patent by Christ et al. (WO 9814436) claims a cyclo
amide such as compound A as inhibitors of HIV reverse
transcriptase. Other prior art includes pyridazine cyclo amide such
as compound B by Turck et al. (Tetrahedron, 49(3), 599-606 (1993))
and compounds such as C by Canonne et al. (J. heterocyclic Chem.
26, 113 (1989)). No activity data were reported in Turck's and
Canonne' publications. 12
[0022] Regarding cyclic amides, Singh et al. and Kumar et al.
(Singh et al. J. Med. Chem. 37(2), 248-254(1994); Kumar et al. J.
Org. Chem. 57(25), 6995-6998 (1992)) disclose compounds such as D
and E claimed as cAMP PDE III inhibitors. 13
DESCRIPTION OF THE INVENTION
[0023] This invention provides compounds of Formula I: 14
[0024] wherein:
[0025] A, B and D are N or CH, with the proviso that A, B and D can
not all be CH;
[0026] R.sup.1 and R.sup.2 are independent substituents selected
from H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to C.sub.6
alkyl, C.sub.2 to C.sub.6 alkenyl, substituted C.sub.2 to C.sub.6
alkenyl, C.sub.2 to C.sub.6 alkynyl, substituted C.sub.2 to C.sub.6
alkynyl, C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3 to
C.sub.8 cycloalkyl, aryl, substituted aryl, heterocyclic,
substituted heterocyclic, COR.sup.A, NR.sup.BCOR.sup.A;
[0027] or R.sup.1 and R.sup.2 are fused to form a spirocyclic ring
selected from a), b) or c), each spirocyclic ring optionally
substituted by from 1 to 3 C.sub.1-C.sub.3 alkyl groups:
[0028] a) a 3 to 8 membered spirocyclic alkyl ring;
[0029] b) a 3 to 8 membered spirocyclic alkenyl ring; or
[0030] c) an optionally substituted 3 to 8 membered heterocyclic
ring containing one to three heteroatoms from the group including
O, S and N; the spirocyclic rings of a), b) and c) being optionally
substituted by from 1 to 4 groups selected from fluorine, C.sub.1
to C.sub.6 alkyl, C.sub.1 to C.sub.6 alkoxy, C.sub.1 to C.sub.6
thioalkyl, --CF.sub.3, --OH, --CN, NH.sub.2, --NH(C.sub.1 to
C.sub.6 alkyl), or --N(C.sub.1 to C.sub.6 alkyl).sub.2;
[0031] R.sup.A is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1
to C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3
alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl;
[0032] R.sup.B is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl;
[0033] R.sup.3 is H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl,
substituted C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.6 alkenyl,
substituted C.sub.1 to C.sub.6 alkenyl, or COR.sup.C;
[0034] R.sup.C is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1
to C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3
alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl;
[0035] R.sup.4 is a trisubstituted benzene ring containing the
substituents X, Y and Z as shown below: 15
[0036] X is taken from the group including halogen, CN, C.sub.1 to
C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl, C.sub.1 to
C.sub.3 alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to
C.sub.3 thioalkoxy, substituted C.sub.1 to C.sub.3 thioalkoxy,
amino, C.sub.1 to C.sub.3 aminoalkyl, substituted C.sub.1 to
C.sub.3 aminoalkyl, NO.sub.2, C.sub.1 to C.sub.3 perfluoroalkyl, 5
or 6 membered heterocyclic ring containing 1 to 3 heteroatoms,
COR.sup.D, OCOR.sup.D, or NR.sup.ECOR.sup.D;
[0037] R.sup.D is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1
to C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3
alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl;
[0038] R.sup.E is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl;
[0039] Y and Z are independent substituents taken from the group
including H, halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.3 alkyl, or C.sub.1 to C.sub.3 thioalkoxy; or
[0040] R.sup.4 is a five or six membered ring with 1, 2, or 3
heteroatoms from the group including O, S, SO, SO.sub.2 or NR.sup.5
and containing one or two independent substituents from the group
including H, halogen, CN, NO.sub.2 and C.sub.1 to C.sub.3 alkyl,
C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3 aminoalkyl,
COR.sup.F, or NR.sup.GCOR.sup.F;
[0041] R.sup.F is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1
to C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3
alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl;
[0042] R.sup.G is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl;
[0043] R.sup.5 is H or C.sub.1 to C.sub.3 alkyl;
[0044] Q is O, S, NR.sup.6, or CR.sup.7R.sup.8;
[0045] R.sub.6 is from the group including CN, C.sub.1 to C.sub.6
alkyl, substituted C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8
cycloalkyl, substituted C.sub.3 to C.sub.8 cycloalkyl, aryl,
substituted aryl, heterocyclic, substituted heterocyclic, or
SO.sub.2CF.sub.3;
[0046] R.sup.7 and R.sup.8 are independent substituents from the
group including H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to
C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3
to C.sub.8 cycloalkyl, aryl, substituted aryl, heterocyclic,
substituted heterocyclic, NO.sub.2, CN, or CO.sub.2R.sup.9;
[0047] R.sup.9 is C.sub.1 to C.sub.3 alkyl;
[0048] or CR.sup.7R.sup.8 form a six membered ring of the structure
below: 16
[0049] W is O or a chemical bond.
[0050] When W is a chemical bond, it is understood that Formula I
exists as: 17
[0051] Preferred compounds are those of Formula I 18
[0052] wherein:
[0053] A, B and D are N or CH, with the proviso that A, B and D can
not all be CH;
[0054] R.sup.1 is H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1
to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl, substituted
C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, COR.sup.A, or
NR.sup.BCOR.sup.A;
[0055] R.sup.2 is H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1
to C.sub.6 alkyl, C.sub.2 to C.sub.6 alkenyl, substituted C.sub.2
to C.sub.6 alkenyl, C.sub.3 to C.sub.8 cycloalkyl, substituted
C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, COR.sup.A, or
NR.sup.BCOR.sup.A; or
[0056] R.sup.1 and R.sup.2 are fused to form the optionally
substituted 3 to 8 membered spirocyclic alkyl, alkenyl or
heterocyclic rings described above;
[0057] R.sup.A is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1
to C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3
alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl;
[0058] R.sup.B is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl;
[0059] R.sup.3 is H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl,
substituted C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.6 alkenyl,
substituted C.sub.1 to C.sub.6 alkenyl, or COR.sup.C;
[0060] R.sup.C is H, C.sub.1 to C.sub.4 alkyl, substituted C.sub.1
to C.sub.4 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.4
alkoxy, substituted C.sub.1 to C.sub.4 alkoxy, C.sub.1 to C.sub.4
aminoalkyl, or substituted C.sub.1 to C.sub.4 aminoalkyl;
[0061] R.sup.4 is a trisubstituted benzene ring containing the
substituents X, Y and Z as shown below: 19
[0062] X is selected from halogen, CN, C.sub.1 to C.sub.3 alkyl,
substituted C.sub.1 to C.sub.3 alkyl, C.sub.1 to C.sub.3 alkoxy,
substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
thioalkoxy, substituted C.sub.1 to C.sub.3 thioalkoxy, C.sub.1 to
C.sub.3 aminoalkyl, substituted C.sub.1 to C.sub.3 aminoalkyl,
NO.sub.2, C.sub.1 to C.sub.3 perfluoroalkyl, a 5 membered
heterocyclic ring containing 1 to 3 heteroatoms, COR.sup.D,
OCOR.sup.D, or NR.sup.ECOR.sup.D;
[0063] R.sup.D is H, C.sub.1 to C.sub.3 alkyl, substituted C.sub.1
to C.sub.3 alkyl, aryl, substituted aryl, C.sub.1 to C.sub.3
alkoxy, substituted C.sub.1 to C.sub.3 alkoxy, C.sub.1 to C.sub.3
aminoalkyl, or substituted C.sub.1 to C.sub.3 aminoalkyl;
[0064] R.sup.E is H, C.sub.1 to C.sub.3 alkyl, or substituted
C.sub.1 to C.sub.3 alkyl;
[0065] Y and Z are independent substituents selected from H,
halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkoxy, C.sub.1 to
C.sub.3 alkyl, or C.sub.1 to C.sub.3 thioalkoxy; or
[0066] R.sup.5 is a five or six membered ring with 1, 2, or 3
heteroatoms from the group including O, S, SO, SO.sub.2 or NR.sup.5
and containing one or two independent substituents from the group
including H, halogen, CN, NO.sub.2 and C.sub.1 to C.sub.3 alkyl, or
C.sub.1 to C.sub.3 alkoxy;
[0067] R.sup.5 is H or C.sub.1 to C.sub.3 alkyl;
[0068] Q is O, S, NR.sup.6, or CR.sup.7R.sup.8;
[0069] R.sup.6 is selected from CN, C.sub.1 to C.sub.6 alkyl,
substituted C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8
cycloalkyl, substituted C.sub.3 to C.sub.8 cycloalkyl, aryl,
substituted aryl, heterocyclic, substituted heterocyclic, or
SO.sub.2CF.sub.3;
[0070] R.sup.7 and R.sup.8 are independent substituents selected
from the group including H, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl,
substituted C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, NO.sub.2, CN, or
CO.sub.2R;
[0071] R.sup.9 is C.sub.1 to C.sub.3 alkyl;
[0072] or CR.sup.7R.sup.8 form a six membered ring of the structure
below: 20
[0073] W is O or a chemical bond;
[0074] or a pharmaceutically acceptable salt thereof.
[0075] Still, more preferred compounds are those of Formula I
21
[0076] wherein:
[0077] A, B and D are N or CH, with the proviso that A, B and D
cannot all be CH;
[0078] R.sup.1=R.sup.2 and are selected from the group of C.sub.1
to C.sub.3 alkyl, substituted C.sub.1 to C.sub.3 alkyl, or
spirocyclic alkyl constructed by fusing R.sup.1 and R.sup.2 to form
a 3 to 6 membered spirocyclic ring;
[0079] R.sup.3 is H, OH, NH.sub.2, C.sub.1 to C.sub.6 alkyl,
substituted C.sub.1 to C.sub.6 alkyl, or COR.sup.C;
[0080] R.sup.C is H, C.sub.1 to C.sub.4 alkyl, or C.sub.1 to
C.sub.4 alkoxy;
[0081] R.sup.4 is a disubstituted benzene ring containing the
substituents X, and Y as shown below: 22
[0082] X is selected from the group of halogen, CN, C.sub.1 to
C.sub.3 alkoxy, C.sub.1 to C.sub.3 alkyl, NO.sub.2, C.sub.1 to
C.sub.3 perfluoroalkyl, 5 membered heterocyclic ring containing 1
to 3 heteroatoms, or C.sub.1 to C.sub.3 thioalkoxy;
[0083] Y is a substituent on the 4' or 5'position from the group
including H, halogen, CN, NO.sub.2, C.sub.1 to C.sub.3 alkoxy,
C.sub.1 to C.sub.4 alkyl, or C.sub.1 to C.sub.3 thioalkoxy; or
[0084] R.sup.4 is a five membered ring with the structure: 23
[0085] R.sup.5 is H, or C.sub.1 to C.sub.3 alkyl, or C.sub.1 to
C.sub.4 CO.sub.2alkyl;
[0086] X' is selected from halogen, CN, NO.sub.2, C.sub.1 to
C.sub.3 alkyl, or C.sub.1 to C.sub.3 alkoxy;
[0087] Y' is selected from H and C.sub.1 to C.sub.4 alkyl; or
24
[0088] wherein:
[0089] A, B and D are N or CH, with the proviso that A, B and D can
not all be CH;
[0090] R.sup.1=R.sup.2 and are selected from CH.sub.3 and
spirocyclic alkyl constructed by fusing R.sup.1 and R.sup.2 to form
a 6 membered spirocyclic ring;
[0091] R.sup.3 is H, OH, NH.sub.2, CH.sub.3, substituted methyl, or
COR.sup.C;
[0092] R.sup.C is H, C.sub.1 to C.sub.3 alkyl, or C.sub.1 to
C.sub.4 alkoxy;
[0093] R.sup.4 is a disubstituted benzene ring containing the
substituents X and Y as shown below: 25
[0094] X is halogen, CN, methoxy, NO.sub.2, or 2-thiazole;
[0095] Y is a substituent on the 4' or 5'position selected from H
and F; or
[0096] R.sup.4is a five membered ring of the structure: 26
[0097] U is O, S, or NH;
[0098] X' is halogen, CN, or NO.sub.2;
[0099] Y' is H or C.sub.1 to C.sub.4 alkyl;
[0100] R.sup.4 is a six membered ring with the structure: 27
[0101] X.sup.1 is N or CX.sup.2;
[0102] X.sup.2 is halogen, CN or NO.sub.2;
[0103] Q is O, S, NR.sup.6, or CR.sup.7R.sup.8;
[0104] R.sub.6 is selected from CN, C.sub.1 to C.sub.6 alkyl,
substituted C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8
cycloalkyl, substituted C.sub.3 to C.sub.8 cycloalkyl, aryl,
substituted aryl, heterocyclic, substituted heterocyclic, or
SO.sub.2CF.sub.3;
[0105] R.sup.7 and R.sup.8 are independent substituents selected
from H, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1 to C.sub.6
alkyl, C.sub.3 to C.sub.8 cycloalkyl, substituted C.sub.3 to
C.sub.8 cycloalkyl, aryl, substituted aryl, heterocyclic,
substituted heterocyclic, NO.sub.2, CN, or CO.sub.2R.sup.9;
[0106] R.sup.9 is C.sub.1 to C.sub.3 alkyl;
[0107] or CR.sup.7R.sup.8 form a six membered ring of the structure
below: 28
[0108] or a pharmaceutically acceptable salt thereof.
[0109] Further preferred compounds include those of Formula I:
[0110] Q is O, S, NR.sup.6, or CR.sup.7R.sup.8;
[0111] R.sub.6 is CN, C.sub.1 to C.sub.6 alkyl, substituted C.sub.1
to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl, substituted
C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, or SO.sub.2CF.sub.3;
[0112] R.sup.7 and R.sup.8 are independent substituents selected
from the group including H, C.sub.1 to C.sub.6 alkyl, substituted
C.sub.1 to C.sub.6 alkyl, C.sub.3 to C.sub.8 cycloalkyl,
substituted C.sub.3 to C.sub.8 cycloalkyl, aryl, substituted aryl,
heterocyclic, substituted heterocyclic, NO.sub.2, CN, or
CO.sub.2R.sup.9;
[0113] R.sup.9 is C.sub.1 to C.sub.3 alkyl;
[0114] or CR.sup.7R.sup.8 form a six membered ring of the
structure: 29
[0115] W is O or a chemical bond;
[0116] or a pharmaceutically acceptable salt thereof
[0117] Each of the generic and subgeneric groups of compounds
described above, as well as the methods of treatment and
pharmaceutical compositions utilizing them, may be divided into two
further subgeneric groups, one in which Q is oxygen and another in
which Q is sulfur or NR.sup.6 or CR.sup.7R.sup.8.
[0118] The compounds of this invention may contain an asymmetric
carbon atom and some of the compounds of this invention may contain
one or more asymmetric centers and may thus give rise to optical
isomers and diastereomers. The shown without respect to
stereochemistry in Formula I, the present invention includes such
optical isomers and diastereomers; as well as the racemic and
resolved, enantiomerically pure R and S stereoisomers; as well as
other mixtures of the R and S stereoisomers and pharmaceutically
acceptable salts thereof.
[0119] The term "alkyl" is used herein to refer to both straight-
and branched-chain saturated aliphatic hydrocarbon groups having
one to eight carbon atoms, preferably one to six carbon atoms;
"alkenyl" is intended to include both straight- and branched-chain
alkyl group with at least one carbon-carbon double bond and two to
eight carbon atoms, preferably two to six carbon atoms; "alkynyl"
group is intended to cover both straight- and branched-chain alkyl
group with at least one carbon-carbon triple bond and two to eight
carbon atoms, preferably two to six carbon atoms.
[0120] The terms "substituted alkyl", "substituted alkenyl", and
"substituted alknyl" refer to alkyl, alkenyl, and alkynyl as just
described having one or more substituents from the group including
halogen, CN, OH, NO.sub.2, amino, aryl, heterocyclic, substituted
aryl, substituted heterocyclic, alkoxy, aryloxy, substituted
alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, arylthio. These
substituents may be attached to any carbon of an alkyl, alkenyl, or
alkynyl group provided that the attachment constitutes a stable
chemical moiety.
[0121] The term "aryl" is used herein to refer to an aromatic
system which may be a single ring or multiple aromatic rings fused
or linked together as such that at least one part of the fused or
linked rings forms the conjugated aromatic system The aryl groups
include but are not limited to phenyl, naphthyl, biphenyl, anthryl,
tetrahydronaphthyl, and phenanthryl.
[0122] The term "substituted aryl" refers to aryl as just defined
having one to four substituents from the group including halogen,
CN, OH, NO.sub.2, amino, alkyl, cycloalkyl, alkenyl, alkynyl,
alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl, alkylcarboxy,
alkylamino, or arylthio.
[0123] The term "heterocyclic" is used herein to describe a stable
4- to 7-membered monocyclic or a stable multicyclic heterocyclic
ring which is saturated, partially unsaturated, or unsaturated, and
which consists of carbon atoms and from one to four heteroatoms
selected from the group including N, O, and S atoms. The N and S
atoms may be oxidized. The heterocyclic ring also includes any
multicyclic ring in which any of the above defined heterocyclic
rings is fused to an aryl ring. The heterocyclic ring may be
attached at any heteroatom or carbon atom provided the resultant
structure is chemically stable. Such heterocyclic groups include,
for example, tetrahydrofuran, piperidinyl, piperazinyl,
2-oxopiperidinyl, azepinyl, pyrrolidinyl, imidazolyl, pyridyl,
pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, isoxazolyl,
morpholinyl, indolyl, quinolinyl, thienyl, furyl, benzofuranyl,
benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, and
isoquinolinyl.
[0124] The term "substituted heterocyclic" is used herein to
describe the heterocyclic just defined having one to four
substituents selected from the group which includes halogen, CN,
OH, NO.sub.2, amino, alkyl, substituted alkyl, cycloalkyl, alkenyl,
substituted alkenyl, alkynyl, alkoxy, aryloxy, substituted
alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, or arylthio. The
term "alkoxy" is used herein to refer to the OR group, where R is
an alkyl or substituted alkyl. The term "aryloxy" is used herein to
refer to the OR group, where R is an aryl or substituted aryl. The
term "alkylcarbonyl" is used herein to refer to the RCO group,
where R is an alkyl or substituted alkyl. The term "alkylcarboxy"
is used herein to refer to the COOR group, where R is an alkyl or
substituted alkyl. The term "aminoalkyl" refers to both secondary
and tertiary amines wherein the alkyl or substituted alkyl groups
contain one to eight carbon atoms, which may be either same or
different and the point of attachment is on the nitrogen atom
"Halogen" refers to Cl, Br, F, or I.
[0125] The compounds of the present invention can be used in the
form of salts derived from pharmaceutically or physiologically
acceptable acids or bases. These salts include, but are not limited
to, the following salts with inorganic acids such as hydrochloric
acid, sulfuric acid, nitric acid, phosphoric acid and, as the case
may be, such organic acids as acetic acid, oxalic acid, succinic
acid, and maleic acid. Other salts include salts with alkali metals
or alkaline earth metals, such as sodium, potassium, calcium or
magnesium in the form of esters, carbamates and other conventional
"pro-drug" forms, which, when administered in such form, convert to
the active moiety in vivo.
[0126] The compounds of this invention have been shown to act as
competitive inhibitors of progesterone binding to the PR and act as
agonists and/or antagonists in functional models, either/or
in-vitro and in-vivo. These compounds may be used for
contraception, in the treatment of fibroids, endometriosis, breast,
uterine, ovarian and prostate cancer, and post menopausal hormone
replacement therapy.
[0127] This invention includes pharmaceutical compositions and
treatments which comprise administering to a mammal a
pharmaceutically effective amount of one or more compounds as
described above wherein Q is oxygen as antagonists of the
progesterone receptor. The invention further provides comparable
methods and compositions which utilize one or more compounds herein
wherein Q is S, NR.sup.6, or CR.sup.7R.sup.8 as agonists of the
progesterone receptor.
[0128] The progesterone receptor antagonists of this invention,
used alone or in combination, can be utilized in methods of
contraception and the treatment and/or prevention of benign and
malignant neoplastic disease. Specific uses of the compounds and
pharmaceutical compositions of invention include the treatment
and/or prevention of uterine myometrial fibroids, endometriosis,
benign prostatic hypertrophy; carcinomas and adenocarcinomas of the
endometrium, ovary, breast, colon, prostate, pituitary, meningioma
and other hormone-dependent tumors. Additional uses of the present
progesterone receptor antagonists include the synchronization of
the estrus in livestock.
[0129] The progesterone receptor agonists of this invention, used
alone or in combination, can be utilized in methods of
contraception and the treatment and/or prevention of dysfunctional
bleeding, uterine leiomyomata, endometriosis; polycystic ovary
syndrome, carcinomas and adenocarcinomas of the endometrium, ovary,
breast, colon, prostate. Additional uses of the invention include
stimulation of food intake.
[0130] When the compounds are employed for the above utilities,
they may be combined with one or more pharmaceutically acceptable
carriers or excipients, for example, solvents, diluents and the
like, and may be administered orally in such forms as tablets,
capsules, dispersible powders, granules, or suspensions containing,
for example, from about 0.05 to 5% of suspending agent, syrups
containing, for example, from about 10 to 50% of sugar, and elixirs
containing, for example, from about 20 to 50% ethanol, and the
like, or parenterally in the form of sterile injectable solutions
or suspensions containing from about 0.05 to 5% suspending agent in
an isotonic medium Such pharmaceutical preparations may contain,
for example, from about 25 to about 90% of the active ingredient in
combination with the carrier, more usually between about 5% and 60%
by weight.
[0131] The effective dosage of active ingredient employed may vary
depending on the particular compound employed, the mode of
administration and the severity of the condition being treated.
However, in general, satisfactory results are obtained when the
compounds of the invention are administered at a daily dosage of
from about 0.5 to about 500 mg/kg of animal body weight, preferably
given in divided doses two to four times a day, or in a sustained
release form For most large mammals, the total daily dosage is from
about 1 to 100 mg, preferably from about 2 to 80 mg. Dosage forms
suitable for internal use comprise from about 0.5 to 500 mg of the
active compound in intimate admixture with a solid or liquid
pharmaceutically acceptable carrier. This dosage regimen may be
adjusted to provide the optimal therapeutic response. For example,
several divided doses may be administered daily or the dose may be
proportionally reduced as indicated by the exigencies of the
therapeutic situation.
[0132] These active compounds may be administered orally as well as
by intravenous, intramuscular, or subcutaneous routes. Solid
carriers include starch, lactose, dicalcium phosphate,
microcrystalline cellulose, sucrose and kaolin, while liquid
carriers include sterile water, polyethylene glycols, non-ionic
surfactants and edible oils such as corn, peanut and sesame oils,
as are appropriate to the nature of the active ingredient and the
particular form of administration desired. Adjuvents customarily
employed in the preparation of pharmaceutical compositions may be
advantageously included, such as flavoring agents, coloring agents,
preserving agents, and antioxidants, for example, vitamin E,
ascorbic acid, BHT and BHA.
[0133] The preferred pharmaceutical compositions from the
standpoint of ease of preparation and administration are solid
compositions, particularly tablets and hard-filled or liquid-filled
capsules. Oral administration of the compounds is preferred.
[0134] These active compounds may also be administered parenterally
or intraperitoneally. Solutions or suspensions of these active
compounds as a free base or pharmacologically acceptable salt can
be prepared in water suitably mixed with a surfactant such as
hydroxypropylcellulose. Dispersions can also be prepared in
glycerol, liquid, polyethylene glycols and mixtures thereof in
oils. Under ordinary conditions of storage and use, these
preparations contain a preservative to prevent the growth of
microorganisms.
[0135] The pharmaceutical forms suitable for injectable use include
sterile aqueous solutions or dispersions and sterile powders for
the extemporaneous preparation of sterile injectable solutions or
dispersions. In all cases, the form must be sterile and must be
fluid to the extent that easy syringe ability exits. It must be
stable under conditions of manufacture and storage and must be
preserved against the contaminating action of microorganisms such
as bacterial and fungi. The carrier can be a solvent or dispersion
medium containing, for example, water, ethanol (e.g., glycerol,
propylene glycol and liquid polyethylene glycol), suitable mixtures
thereof, and vegetable oil.
[0136] The compounds of this invention can be prepared following
the Schemes illustrated below:
[0137] As demonstrated in Scheme I, the compounds of this invention
are generally prepared by employing the suitable coupling reaction
as a final step. An appropriately substituted ortho-amino acid or
its derivatives such as an ethyl ester (X=Br, I, Cl, or a latent
coupling precursor such as alkoxy group which can be converted into
an OTf group suitable in the coupling reaction) is treated with a
suitable organo metallic reagent, e.g. Grignard reagent, in
appropriate nonprotic solvents which include but not limited to THF
or ether to give ortho-amino carbinol 2 under an inert atmosphere
such as argon or nitrogen at -78.degree. C. to room temperature.
Ring closure of carbinol 2 to yield oxazin-2-ones 3 is commonly
effected by a condensing agent such as carbonyldimidazole,
phosgene, dimethylcarbonate, or diethylcarbonate in a suitable
nonprotic solvent such as THF at temperatures ranging from room
temperature to 65.degree. C. The arylation of oxazin-2-ones 3 to
yield 4 can be effected by various coupling reactions including
Suzuki, Stille reactions etc. These reactions are commonly
performed in the presence of a transition metallic catalyst, e.g.,
palladium or nickel complex often with phosphino ligands, e.g.,
Ph.sub.3P, 1,1'-bis(diphenylphosphino)ferrocene,
1,2-bis(diphenylphosphino)ethane or palladium salt such as
palladium acetate. Under this catalytic condition, an appropriately
substituted nucleophilic reagent, e.g., aryl boronic acid,
arylstannane, or aryl zinc compound, is coupled with oxazinones 3
to give compounds 4. If a base is needed in the reaction, the
commonly used bases include but not limited to sodium bicarbonate,
sodium carbonate, potassium phosphate, barium carbonate, potassium
acetate, or cesium fluoride. The most commonly used solvents in
these reactions include benzene, DMF, isopropanol, ethanol, DME,
ether, acetone or a mixture of any one of these solvent and water.
The coupling reaction is generally executed under an inert
atmosphere such as nitrogen or argon at temperatures ranging from
room temperature to 95.degree. C. Oxazinones 3 can be converted
into a nucleophile such as boronic acid which can be coupled with
an appropriate electrophile, e.g., aryl bromide or aryl iodide, to
yield 6 employing the coupling reaction condition as described
above. 30
[0138] The transformation of 3 into 5 can be effected by treating 3
with an organo metallic reagent, e.g., n-BuLi, in a nonprotic
solvent such as THF or ether followed by quenching the reaction
solution with a suitable electrophile such as trimethyl borate,
triisopropyl borate, bishexalkyl tin reagent, or zinc chloride at
temperatures ranging from -78.degree. C. to room temperature under
an inert atmosphere such as argon or nitrogen. Conversion of
carbamate 6 to thiocarbamate 7 can be readily effected by treatment
of 6 with a suitable sulfur reagent such as P.sub.2S.sub.5 or
Lawesson's reagent in a suitable nonprotic solvent such as toluene,
chlorobenzene, benzene, or xylene under an inert atmosphere such as
argon or nitrogen at the temperature of boiling solvent.
[0139] Scheme II describes the procedures to prepare oxazinones
bearing two different substituents at position-4. The Weinreb amide
9 can be prepared from an appropriately substituted isatoic
anhydride when treated with N-, O-dimethylhydroxylamine
hydrochloride salt in a protic solvent such as ethanol or
isopropanol at reflux under an inert atmosphere such as argon or
nitrogen. Coupling of amide 9 with an aryl electrophile such as
aryl boronic acid or arylstannane to give 10 can be effected by
employing a typical coupling reaction such as Suzuki, Stille
coupling procedure in a similar fashion as described for the
preparation of oxazinones 4. Treatment of Weinreb amide 10 with
organo metallic compounds, e.g., alkyllithium, alkynyllithium,
aryllithium, or their Grignard counterpart in a nonprotic solvent
such as THF or ether under an inert atmosphere such as argon or
nitrogen at -78 .degree. to room temperature affords amino ketone
11. Conversion of ketone 11 to carbinol 12 can be effected by
treatment of 10 with an organo metallic reagent such as alkyl,
alkynyl, or aryl Grignard compound in a nonprotic solvent such as
THF or ether under an inert atmosphere such as argon or nitrogen at
-78.degree. C. to room temperature. Conversion of ketone 11 to
carbinol 12 can also be effected by reduction of ketone group of 11
to the carbinol moiety of 12 using an appropriate reducing reagent
such as lithium aluminum hydride, sodium borohydride in a suitable
solvent such as THF, ether, or anhydrous alcohol under an inert
atmosphere in the temperature ranging from 0.degree. C. to the
boiling point of the solvent. Ring closure of carbinol 12 to
produce the compounds of this invention, 13, can be accomplished
with condensing agents such as carbonyldiimidazole, phosgene,
dimethylcarbonate, or diethylcarbonate in a suitable nonprotic
solvent such as THF at temperatures ranging from room temperature
to 65.degree. C. Conversion of carbamate 13 to thiocarbamate 14 can
be readily effected by treatment of 13 with a suitable sulfur
reagent such as P.sub.2S.sub.5 or Lawesson's reagent in a suitable
nonprotic solvent such as toluene, chlorobenzene, benzene, or
xylene under an inert atmosphere such as argon or nitrogen at the
temperature of boiling solvent. 31
[0140] Alternatively, ortho-amino ketone 11 can be prepared by
treatment of ortho-amino nitrile 16 with an organo metallic
compound such as an organo lithium reagent or Grignard reagent in a
suitable solvent such as THF or ether under an inert atmosphere
such as argon or nitrogen at temperatures ranging from -78.degree.
C. to room temperature as illustrated in Scheme III. Nitrile 16 can
be readily prepared from an appropriately substituted nitrile such
as bromobenzonitrile 15 using a suitable coupling reaction such as
Stille or Suzuki protocol carried out in a similar fashion as
described for the preparation of the Weinreb amide 10. 32
[0141] The following non-limiting examples illustrate the compounds
of the invention.
EXAMPLE 1
2-Amino-5-bromo-3-pyridine Carboxylic Acid
[0142] To a solution of 2-amino-nicotinic acid (10 g, 72.5 mmol) in
acetic acid (70 mL) was dropwise added bromine (9.8 mL, 79.8 mmol)
at room temperature under nitrogen. Upon completion of the
reaction, the solvent was removed in vacuo, the residue triturated
with ether and collected on a filter to give
2-amino-5-bromo-3-pyridine carboxylic acid as a yellow solid (15.7
g, 99%): mp 272.degree. C., (decomposed); .sup.1H-NMR
(DMSO-d.sub.6) .delta. 8.8-7.8 (bs, 2H), 8.44 (d, 1H, J=2.48 Hz),
8.34 (d, 1H, J=2.48 Hz); MS (EI) m/z 216/218 ([M+].sup.+, 100%)
EXAMPLE 2
2-(2-Amino-5-bromo-pyridin-3-yl)-propan-2-ol
[0143] To a solution of 2-amino-5-bromo-3-pyridine carboxylic acid
(5 g, 23 mmol) in THF (70 mL) at 0.degree. C. was added methyl
magnesium bromide (13.7 g, 115 mmol) under nitrogen. After
addition, the reaction mixture was heated at 65.degree. C. for 12
hours, cooled to room temperature and quenched with saturated
aqueous ammonium chloride. The ethyl acetate was added and organic
layer was separated, dried over sodium sulfate and concentrated.
The residue was purified via flash chromatography to give
2-(2-amino-5-bromo-pyridin-3-yl)-propan-2-ol as a yellow solid (1.2
g, 23%): mp 107-108.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) 67.89
(d, 1H, J=2.3 Hz), 7.40 (d, 1H, J=2.3 Hz), 6.28 (bs, 2H), 5.51 (s,
1H), 1.4 (s, 6H); MS (APCI) m/z 231 ([M+H].sup.+, 100%).
EXAMPLE 3
6-Bromo-4,4'-dimethyl-1,4-dihydro-3-oxa-1,8-diaza-naphthalen-2-one
[0144] A mixture of 2-(2-amino-5-bromo-pyridin-3-yl)-propan-2-ol
(0.86 g, 3.7 mmol) and 1,1'-carbonyldiimidazole (excess) in THF (10
mL) was heated at 35.degree. C. overnight. The reaction solution
was cooled to room temperature, poured into ethyl acetate (200 mL),
washed with 1 N HCl (2.times.50 mL), dried over sodium sulfate, and
concentrated. The residue was purified by a flash chromatography
(silica gel, 25% ethyl acetate/hexane) to afford
6-bromo-4,4'-dimethyl-1,4-dihydro-3-oxa-1,8-dia-
za-naphthalen-2-one (0.9 g, 94%) as a white solid: mp
251-252.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 10.9 (s,
1H), 8.32 (d, 1H, J=2.19 Hz), 8.0 (d, 1H, J=2.22 Hz), 1.6 (s, 6H);
MS (EI) m/z 256 ([M].sup.+, 80%); Anal. Calc. For
C.sub.9H.sub.9BrN.sub.2O.sub.2: C, 42.05, H, 3.53, N, 10.90. Found:
C, 42.15, H, 3.65, N, 10.80.
EXAMPLE 4
6-(3-Chloro-phenyl)-4,4-dimethyl-1,4-dihydro-3-oxa-1,8-diaza-naphthalene-2-
-one
[0145] A mixture of
6-bromo-4,4'-dimethyl-1,4-dihydro-3-oxa-1,8-diaza-naph-
thalen-2-one (0.1 g, 0.39 mmol), 3-chlorophenyl boronic acid (0.075
g, 0.47 mmol), tetrakis(triphenylphosphine)-palladium (0) (0.023 g,
0.02 mmol), and sodium carbonate (0.1 g, 0.94 mmol) in DME (8 mL)
and water (5 mL) was subject to a blanket of nitrogen for 15
minutes at 50.degree. C. and then was heated at 85.degree. C. for
30 minutes. The reaction was cooled to room temperature and ethyl
acetate (100 mL) was added. The organic layer washed with aqueous
ammonium chloride (2.times.50 mL) and with brine (50 mL), dried
over magnesium sulfate and concentrated. The product was dissolved
in dichloromethane and passed through a plug of magnesol. The
solvent was removed and the clear oil obtained triturated with
ether (25 mL) to give
6-(3-chloro-phenyl)-4,4-dimethyl-1,4-dihydro-3-
-oxa-1,8-diaza-naphthalene-2-one as a white solid (0.087 g, 80%):
mp 214-215.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 10.9 (s,
1H), 8.56 (d, 1H, J=2.35 Hz), 8.06 (d, 1H, J=2.35 Hz), 7.86 (t, 1H,
J=2.35 Hz), 7.72 (td, 1H, J=9.4, 2.35 Hz), 7.54-7.44 (m, 2H), 1.69
(s, 6H); MS (ESI) m/z 289 ([M+H].sup.+, 100%); Anal. Calc. For
C.sub.17H.sub.17NO.sub.3: C, 62.40, H, 4.54, N, 9.70. Found: C,
60.53, H, 4.40, N, 9.21.
EXAMPLE 5
6-Chloro-3-nitro-pyridine-2-carbonitile
[0146] A mixture of 2,6-dichloro-3-nitropyridine and cuprous
cyanide in 1-methyl-2-pyrrolidinone was quickly heated to
180.degree. C. and maintained at that temperature for 15 minutes
with vigorous stirring. The mixture was cooled to -10.degree. C.
and the deep brown solution was poured into ice water (3.5 L) and
stirred for 30 min. The flocculent brown precipitate was collected
and washed with H.sub.2O. After drying for about 1.5 h, the moist
solid was extracted with boiling toluene (3.times.300 mL). The
combined toluene extracts were washed with H.sub.2O, brine, and
dried (MgSO.sub.4), concentrated. The crude product was
crystallized from EtOAc/hexane to afford the title compound: mp
115-117.degree. C.; .sup.1H NMR(DMSO-d.sub.6) .delta. 8.16 (dd, 1H,
J=8.7, 3.0 Hz), 8.82 (d, 1H, J=9.0 Hz); MS (EI) m/z 183/185
(M+H).sup.+; Anal. Calc. For C.sub.6H.sub.2ClN.sub.3O.sub.2: C,
39.26, H, 1.10, N, 22.89. Found: C, 39.47, H, 1.38, N, 22.77.
EXAMPLE 6
3-Amino-6-chloro-pyridine-2-carbonitrile
[0147] To a solution of 2,6-dichloro-3-nitropyridine(4.8 g, 26.15
mmol) in MeOH (60 mL) and concentrated HCl (25 mL) was slowly added
iron powder (5.12g, 91.53 mmol). After the completion of addition,
the mixture was refluxed for 45 minutes and poured into 700 mL of
H.sub.2O. Filtration of the resulting slurry gave a dull yellow
solid. The filtrate was made basic with concentrated NH.sub.4OH,
the resulting slurry was filtered and both the solid and the
filtrates were extracted with ether. The combined extracts were
dried (MgSO.sub.4) and evaporated to give the title compound as a
creamy solid (2.8 g, 58%): mp 162-165.degree. C. which was used in
next step without further purification.
EXAMPLE 7
1-(3-Amino-6-chloro-pyridin-2-yl)-ethanone
[0148] To a solution of 3-amino-6-chloro-pyridine-2-carbonitrile
(0.75 g, 4.88 mmol) in anhydrous THF (25 mL) under nitrogen was
added a solution of methylmagnesium bromide (3M in ether, 8.1 mL,
24.3 mmol). The reaction mixture was stirred for 30 minutes and
then slowly quenched with H.sub.2O, and treated with 5N HCl
solution. The mixture was extracted with ethyl acetate (3.times.100
mL) and organic extracts were washed with brine, and dried
(MgSO.sub.4). After removal of the solvent, the residue was
purified by a chromatography using EtOAc/hexane mixture (1:3) as
eluent to afford the title compound as a greenish crystalline
solid: (0.71 g, 85%): mp: 108-110.degree. C. .sup.1H NMR
(DMSO-d.sub.6) .delta. 2.51 (s, 3H), 7.28-7.39 (mn, 4H); MS(ESI)
m/z 171/173 (M+H).sup.+; Anal. Calc. For C.sub.7H.sub.7ClN.sub.2O:
C, 49.28, H, 4.14, N, 16.14. Found: C, 49.70, H, 4.03, N,
16.41.
EXAMPLE 8
1-(3-Amino-6-chloro-pyridin-2-yl)-propan-2-ol
[0149] To a solution of 1-(3-amino-6-chloro-pyridin-2-yl)-ethanone
in anhydrous THF under N.sub.2 was added a solution of
methylmagnesium bromide (3N in ether). The resulting reaction
mixture was stirred at room temperature for 4 h, then slowly
quenched with H.sub.2O, treated with 0.5 N HCl and stirred for 30
minutes, diluted with ethyl acetate, washed with brine, dried
(MgSO.sub.4), concentrated, and chromatographed using a mixture of
EtOAc/Hexane (3.5:6.5) to afford the title compound as a white
crystals (0.45 g, 82%): mp: 118-121.degree. C. .sup.1H
NMR(DMSO-d.sub.6) .delta. 1.45(s, 6H), 3.35(s, 1H), 5.51(s, 1H),
5.68(s, 1H), 7.02(s, 1H); MS((+)APCI) m/z 187/189 (M+H).sup.+;
Anal. Calc. For C.sub.8H.sub.11ClN.sub.2O: C, 51.48, H, 5.94, N,
15.01. Found: C, 51.22, H, 5.99, N, 14.47.
EXAMPLE 9
6-Chloro-4,4-dimethyl-1,4-dihydro-3-oxa-1,5-diaza-naphthalen-2-one
[0150] To a solution of
1-(3-amino-6-chloro-3-nitro-pyridin-2-yl)-propan-2- -ol (0.3 g,
1.67 mmol) in anhydrous THF (20 mL) was added a solution of
1,1'-carbonyldimidazole (0.68 g, 4.12 mmol) in anhydrous THF (10
mL). The reaction mixture was heated under reflux for 3 h. The
reaction mixture was treated with 0.5N HCl, washed with brine,
H.sub.2O, dried (MgSO.sub.4), concentrated and crystallized from
EtOAc/hexane to obtain the title compound as a white crystalline
solid (0.2 g, 56%): mp 175-178.degree. C. .sup.1H NMR(DMSO-d.sub.6)
.delta. 1.60 (s, 6H), 7.30 (d, 1H, J=8.41 Hz), 7.41 (d, 1H, J=8.41
Hz), 10.47 (s, 1H); MS((+)APCI) m/z 213 (M+H).sup.+; Anal. Calc.
For C.sub.9H.sub.9ClN.sub.2O.sub.2: C, 50.84, H, 4.27, N, 13.17.
Found: C, 50.99, H, 4.28, N, 12.98.
EXAMPLE 10
[0151] The compounds of this invention were tested in the relevant
assay as described below and their potency are in the range of 0.01
nM to 5 .mu.M in the in vitro assays and 0.001 to 300 mg/kg in the
in vivo assays. The selected example is example 4. 33
[0152] Example 4, hPR CV-1, IC.sub.50=0.8 .mu.M
[0153] A. In-vitro Biology
[0154] The in-vitro biology is determined by (1) competitive
Radioligand Binding: using the A-form of the human progesterone
receptor with progesterone as the radioligand; (2) co-transfection
assay, which provides functional activity expressed as agonist EC50
and Antagonist IC50 values; (3) a T47D cell proliferation, which is
a further functional assay which also provides agonist and
antagonist data; and (4) T47D cell alkaline phosphatase assay,
which is a further functional assay which also provides agonist and
antagonist data.
[0155] 1. hPR Binding Assay
[0156] This assay is carried out in accordance with: Pathirana, C.;
Stein, R. B.; Berger, T. S.; Fenical, W.; Ianiro, T.; Mais, D. E.;
Torres, A.; Glodman, M. E., Nonsteroidal human progesterone
receptor modulators from the marine alga cymoplia barbata, J.
Steroid Biochem. Mol. Biol., 1992, 41, 733-738.
[0157] 2. PRE-luciferase Assay in CV-1 Cells
[0158] The object of this assay is to determine a compound's
progestational or antiprogestational potency based on its effect on
PRE-luciferase reporter activity in CV-1 cells co-transfected with
human PR and PRE-luciferase plasmids. The materials methods used in
the assay are as follows.
[0159] a. Medium:
[0160] The growth medium was as follows: DMEM (BioWhittaker)
containing 10% (v/v) fetal bovine serum (heat inactivated), 0.1 mM
MEM non-essential amino acids, 100 U/ml penicillin, 100 mg/ml
streptomycin, and 2 mM GlutaMax (GIBCO, BRL). The experimental
medium was as follows: DMEM (BioWhittaker), phenol red-free,
containing 10% (v/v) charcoal-stripped fetal bovine serum
(heat-inactivated), 0.1 mM MEM non-essential amino acids, 100 U/ml
penicillin, 100 mg/ml streptomycin, and 2 mM GlutaMax (GIBCO,
BRL).
[0161] b. Cell Culture, Transfection, Treatment and Luciferase
Assay
[0162] Stock CV-1 cells are maintained in growth medium
Co-transfection is done using 1.2.times.10.sup.7 cells, 5 mg pLEM
plasmid with hPR-B inserted at Sph1 and BamH1 sites, 10 mg pGL3
plasmid with two PREs upstream of the luciferase sequence, and 50
mg sonicated calf thymus DNA as carrier DNA in 250 ml.
Electroporation is carried out at 260 V and 1,000 mF in a Biorad
Gene Pulser II. After electroporation, cells are resuspended in
growth medium and plated in 96-well plate at 40,000 cells/well in
200 .mu.l. Following overnight incubation, the medium is changed to
experimental medium. Cells are then treated with reference or test
compounds in experimental medium Compounds are tested for
antiprogestational activity in the presence of 3 nM progesterone.
Twenty-four hr. after treatment, the medium is discarded, cells are
washed three times with D-PBS (GIBCO, BRL). Fifty .mu.l of cell
lysis buffer (Promega, Madison, Wis.) is added to each well and the
plates are shaken for 15 min in a Titer Plate Shaker (Lab Line
Instrument, Inc.). Luciferase activity is measured using luciferase
reagents from Promega.
[0163] c. Analysis of Results:
[0164] Each treatment consists of at least 4 replicates. Log
transformed data are used for analysis of variance and nonlinear
dose response curve fitting for both agonist and antagonist modes.
Huber weighting is used to downweight the effects of outliers.
EC.sub.50 or IC.sub.50 values are calculated from the retransformed
values. JMP software (SAS Institute, Inc.) is used for both one-way
analysis of variance and non-linear response analyses.
[0165] d. Reference Compounds:
[0166] Progesterone and trimegestone are reference progestins and
RU486 is the reference antiprogestin. All reference compounds are
run in full dose-response curves and the EC.sub.50 or IC.sub.50
values are calculated.
1TABLE 1 Estimated EC.sub.50, standard error (SE), and 95%
confidence intervals (CI) for reference progestins from three
individual studies EC50 95% CI Compound Exp. (nM) SE lower upper
Progesterone 1 0.616 0.026 0.509 0.746 2 0.402 0.019 0.323 0.501 3
0.486 0.028 0.371 0.637 Trimegestone 1 0.0075 0.0002 0.0066 0.0085
2 0.0081 0.0003 0.0070 0.0094 3 0.0067 0.0003 0.0055 0.0082
[0167]
2TABLE 2 Estimated IC.sub.50, standard error (SE), and 95%
confident interval (CI) for the antiprogestin, RU486 from three
individual studies IC 50 95% CI Compound Exp. (nM) SE lower upper
RU486 1 0.028 0.002 0.019 0.042 2 0.037 0.002 0.029 0.048 3 0.019
0.001 0.013 0.027
[0168] Progestational Activity:
[0169] Compounds that increase PRE-luciferase activity
significantly (p<0.05) compared to vehicle control are
considered active.
[0170] Antiprogestational Activity:
[0171] Compounds that decrease 3 nM progesterone induced
PRE-luciferase activity significantly (p<0.05)
[0172] EC.sub.50:
[0173] Concentration of a compound that gives half-maximal increase
PRE-luciferase activity (default-nM) with SE.
[0174] IC.sub.50:
[0175] Concentration of a compound that gives half-maximal decrease
in 3 nM progesterone induced PRE-luciferase activity (default-nM)
with SE.
[0176] 3. T47D Cell Proliferation Assay
[0177] The objective of this assay is the determination of
progestational and antiprogestational potency by using a cell
proliferation assay in T47D cells. A compound's effect on DNA
synthesis in T47D cells is measured. The materials and methods used
in this assay are as follows.
[0178] a. Growth Medium:
[0179] DMEM:F12 (1:1) (GIBCO, BRL) supplemented with 10% (v/v)
fetal bovine serum (not heat-inactivated), 100 U/ml penicillin, 100
mg/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL).
[0180] b. Treatment Medium:
[0181] Minimum Essential Medium (MEM) (#51200-038GIBCO, BRL) phenol
red-free supplemented with 0.5% charcoal stripped fetal bovine
serum, 100 U/ml penicillin, 200 mg/ml streptomycin, and 2 mM
GlutaMax (GIBCO, BRL).
[0182] c. Cell Culture
[0183] Stock T47 D cells are maintained in growth medium For BrdU
incorporation assay, cells are plated in 96-well plates (Falcon,
Becton Dickinson Labware) at 10,000 cells/well in growth medium
After overnight incubation, the medium is changed to treatment
medium and cells are cultured for an additional 24 hr before
treatment. Stock compounds are dissolved in appropriate vehicle
(100% ethanol or 50% ethanol/50% DMSO), subsequently diluted in
treatment medium and added to the cells. Progestin and
antiprogestin reference compounds are run in full dose-response
curves. The final concentration of vehicle is 0.1%. In control
wells, cells receive vehicle only. Antiprogestins are tested in the
presence of 0.03 nM trimegestone, the reference progestin agonist.
Twenty-four hours after treatment, the medium is discarded and
cells are labeled with 10 mM BrdU (Amersham Life Science, Arlington
Heights, Ill.) in treatment medium for 4 hr.
[0184] d. Cell Proliferation Assay
[0185] At the end of BrdU labeling, the medium is removed and BrdU
incorporation is measured using a cell proliferation ELISA kit
(#RPN 250, Amersham Life Science) according to manufacturer's
instructions. Briefly, cells are fixed in an ethanol containing
fixative for 30 min, followed by incubation in a blocking buffer
for 30 min to reduce background. Peroxidase-labeled anti-BrdU
antibody is added to the wells and incubated for 60 min. The cells
are rinsed three times with PBS and incubated with
3,3'5,5'-tetramethylbenzidine (TMB) substrate for 10-20 min
depending upon the potency of tested compounds. Then 25 .mu.l of 1
M sulfuric acid is added to each well to stop color reaction and
optical density is read in a plate reader at 450 nm within 5
min.
[0186] e. Analysis of Results:
[0187] Square root-transformed data are used for analysis of
variance and nonlinear dose response curve fitting for both agonist
and antagonist modes. Huber weighting is used to downweight the
effects of outliers. EC.sub.50 or IC.sub.50 values are calculated
from the retransformed values. JMP software (SAS Institute, Inc.)
is used for both one-way analysis of variance and non-linear dose
response analyses in both single dose and dose response
studies.
[0188] f. Reference Compounds:
[0189] Trimegestone and medroxyprogesterone acetate (MPA) are
reference progestins and RU486 is the reference antiprogestin. All
reference compounds are run in full dose-response curves and the
EC.sub.50 or IC.sub.50 values are calculated.
3TABLE 3 Estimated EC.sub.50, standard error (SE), and 95%
confidence intervals (CI) for individual studies EC.sub.50 95% CI
Compound Exp (nM) SE lower upper Trimegestone 1 0.017 0.003 0.007
0.040 2 0.014 0.001 0.011 0.017 3 0.019 0.001 0.016 0.024 MPA 1
0.019 0.001 0.013 0.027 2 0.017 0.001 0.011 0.024
[0190]
4TABLE 4 Estimated IC.sub.50, standard error, and 95% confident
interval for the antiprogestin, RU486 IC.sub.50 95% CI Compound Exp
(nM) SE lower upper RU486 1 0.011 0.001 0.008 0.014 2 0.016 0.001
0.014 0.020 3 0.018 0.001 0.014 0.022
[0191] EC.sub.50: Concentration of a compound that gives
half-maximal increase in BrdU incorporation with SE; IC.sub.50:
Concentration of a compound that gives half-maximal decrease in 0.1
trimegestone induced BrdU incorporation with SE
[0192] 4. T47D Cell Alkaline Phosphatase Assay
[0193] The purpose of this assay is to identify progestins or
antiprogestins by determining a compound's effect on alkaline
phosphatase activity in T47D cells. The materials and methods used
in this assay are as follows.
[0194] a. Culture Medium:
[0195] DMEM:F12 (1:1) (GIBCO, BRL) supplemented with 5% (v/v)
charcoal stripped fetal bovine serum (not heat-inactivated), 100
U/ml penicillin, 100 .mu.g/ml streptomycin, and 2 mM GlutaMax
(GIBCO, BRL).
[0196] b. Alkaline Phosphatase Assay Buffer:
[0197] I. 0.1 M Tris-HCl, pH 9.8, containing 0.2% Triton X-100
[0198] II. 0.1 M Tris-HCl, pH 9.8 containing 4 mM p-nitrophenyl
phosphate (Sigma).
[0199] c. Cell Culture and Treatment:
[0200] Frozen T47D cells were thawed in a 37.degree. C. water bath
and diluted to 280,000 cells/ml in culture medium. To each well in
a 96-well plate (Falcon, Becton Dickinson Labware), 180 .mu.l of
diluted cell suspension was added. Twenty .mu.l of reference or
test compounds diluted in the culture medium was then added to each
well. When testing for progestin antagonist activity, reference
antiprogestins or test compounds were added in the presence of 1 nM
progesterone. The cells were incubated at 37.degree. C. in a 5%
CO.sub.2/humidified atmosphere for 24 hr.
[0201] d. Alkaline Phosphatase Enzyme Assay:
[0202] At the end of treatment, the medium was removed from the
plate and fifty .mu.l of assay buffer I was added to each well. The
plates were shaken in a titer plate shaker for 15 min. Then 150
.mu.l of assay buffer II was added to each well. Optical density
measurements were taken at 5 min intervals for 30 min at a test
wavelength of 405 nM.
[0203] e. Analysis of Results: Analysis of Dose-response Data
[0204] For reference and test compounds, a dose response curve is
generated for dose (X-axis) vs. the rate of enzyme reaction (slope)
(Y-axis). Square root-transformed data are used for analysis of
variance and nonlinear dose response curve fitting for both agonist
and antagonist modes. Huber weighting is used to downweight the
effects of outliers. EC.sub.50 or IC.sub.50 values are calculated
from the retransformed values. JMP software (SAS Institute, Inc.)
is used for both one-way analysis of variance and non-linear dose
response analyses in both single dose and dose response
studies.
[0205] f. Reference Compounds:
[0206] Progesterone and trimegestone are reference progestins and
RU486 is the reference antiprogestin. All reference compounds are
run in full dose response curves and the EC.sub.50 or IC.sub.50
values are calculated.
5TABLE 5 Estimated EC.sub.50, standard error (SE), and 95%
confidence intervals (CI) for reference progestins from three
independent experiments EC50 95% CI Compound Exp. (nM) SE lower
upper Progesterone 1 0.839 0.030 0.706 0.996 2 0.639 0.006 0.611
0.669 3 1.286 0.029 1.158 1.429 Trimegestone 1 0.084 0.002 0.076
0.091 2 0.076 0.001 0.072 0.080 3 0.160 0.004 0.141 0.181
[0207]
6TABLE 6 Estimated IC.sub.50, standard error, and 95% confident
interval for the reference antiprogestin RU486 from three
independent experiments IC 50 95% CI Compound Exp (nM) SE lower
upper RU486 1 0.103 0.002 0.092 0.115 2 0.120 0.001 0.115 0.126 3
0.094 0.007 0.066 0.134
[0208] B. In-vivo Biology
[0209] The primary in-vivo assay is the rat decidualization model,
which may be used to determine progestational effects of both
agonists and antagonists. The secondary in-vivo assay is the rat
ovulation inhibition model, which is under development, and hence
the protocol is un-available.
[0210] 1. Rat Decidualization Assay
[0211] The objective of this procedure is used to evaluate the
effect of progestins and antiprogestins on rat uterine
decidualization and compare the relative potencies of various test
compounds. The materials and methods used in this assay are as
follows.
[0212] a. Methods:
[0213] Test compounds are dissolved in 100% ethanol and mixed with
corn oil (vehicle). Stock solutions of the test compounds in oil
(Mazola.TM.) are then prepared by heating (-80.degree. C.) the
mixture to evaporate ethanol. Test compounds are subsequently
diluted with 100% corn oil or 10% ethanol in corn oil prior to the
treatment of animals. No difference in decidual response was found
when these two vehicles were compared.
[0214] b. Animals (RACUC Protocol #5002)
[0215] Ovariectomized mature female Sprague-Dawley rats (-60-day
old and 230 g) are obtained from Taconic (Taconic Farms, N.Y.)
following surgery. Ovariectomy is performed at least 10 days prior
to treatment to reduce circulating sex steroids. Animals are housed
under 12 hr light/dark cycle and given standard rat chow and water
ad libitum.
[0216] c. Treatment
[0217] Rats are weighed and randomly assigned to groups of 4 or 5
before treatment. Test compounds in 0.2 ml vehicle are administered
by subcutaneous injection in the nape of the neck or by gavage
using 0.5 ml. The animals are treated once daily for seven days.
For testing antiprogestins, animals are given the test compounds
and a EC50 dose of progesterone (5.6 mg/kg) during the first three
days of treatment. Following decidual stimulation, animals continue
to receive progesterone until necropsy four days later.
[0218] d. Dosing
[0219] Doses are prepared based upon mg/kg mean group body weight.
In all studies, a control group receiving vehicle is included.
Determination of dose-response curves is carried out using doses
with half log increases (e.g. 0.1, 0.3, 1.0, 3.0 mg/kg . . . ).
[0220] e. Decidual Induction
[0221] Approximately 24 hr after the third injection,
decidualization is induced in one of the uterine horns by
scratching the antimesometrial luminal epithelium with a blunt 21 G
needle. The contralateral horn is not scratched and serves as an
unstimulated control. Approximately 24 hr following the final
treatment, rats are sacrificed by CO.sub.2 asphyxiation and body
weight measured. Uteri are removed and trimmed of fat. Decidualized
(D-horn) and control (C-horn) uterine horns are weighed
separately.
[0222] f. Analysis of Results:
[0223] The increase in weight of the decidualized uterine horn is
calculated by D-horn/C-horn and logarithmic transformation is used
to maximize normality and homogeneity of variance. The Huber
M-estimator is used to down weight the outlying transformed
observations for both dose-response curve fitting and one-way
analysis of variance. JMP software (SAS Institute, Inc.) is used
for both one-way ANOVA and non-linear dose-response analyses.
[0224] g. Reference Compounds:
[0225] All progestin reference compounds were run in fill
dose-response curves and the EC.sub.50 for uterine wet weight were
calculated.
7TABLE 7 Estimated EC.sub.50, standard error (SE), and 95%
confidence intervals for individual studies EC.sub.50 95% CI
Compound Exp (mg/kg, s.c.) SE lower upper Progesterone 1 5.50 0.77
4.21 7.20 2 6.21 1.12 4.41 8.76 3-Ketodesogestrel 1 0.11 0.02 0.07
0.16 2 0.10 0.05 0.11 0.25 3 0.06 0.03 0.03 0.14 Levonorgestrel 1
0.08 0.03 0.04 0.16 2 0.12 0.02 0.09 0.17 3 0.09 0.02 0.06 0.13 4
0.09 0.02 0.06 0.14 MPA 1 0.42 0.03 0.29 0.60 2 0.39 0.05 0.22 0.67
3 0.39 0.04 0.25 0.61
[0226]
8TABLE 8 Estimated average EC.sub.50, standard error, and 95%
confidence intervals for dose-response curves of 3 reference
compounds EC.sub.50 95% CI Compound (mg/kg. s.c.) SE lower upper
Progesterone 5.62 0.62 4.55 7.00 3-Ketodesogestrel 0.10 0.02 0.07
0.14 Levonorgestrel 0.10 0.01 0.08 0.12
[0227]
9TABLE 9 Estimated IC.sub.50, standard error, and 95% confident
interval for the antiprogestin, RU 486 IC.sub.50 95% CI Compound
Exp. (mg/kg, p.o.) SE lower upper RU 486 1 0.21 0.07 0.05 0.96 2
0.14 0.02 0.08 0.27
[0228] Concentration: Compound concentration in assay(default-mg/kg
body weight)
[0229] Route of administration: Route the compound is administered
to the animals
[0230] Body weight: Mean total animal body weight (default-kg)
[0231] D-horn: Wet weight of decidualized uterine horn
(default-mg)
[0232] C-horn: Wet weight of control uterine horn (default-mg)
[0233] Decidual response: [(D-C)/C].times.100%
[0234] Progestational activity: Compounds that induce
decidualization significantly (p<0.05) compared to vehicle
control are considered active
[0235] Antiprogestational activity: Compounds that decrease
EC.sub.50 progesterone induced decidualization significantly
(p<0.05)
[0236] EC.sub.50 for uterine weight: Concentration of compound that
gives half-maximal increase in decidual response
(default-mg/kg)
[0237] IC.sub.50 for uterine weight: Concentration of compound that
gives half-maximal decrease in EC.sub.50 progesterone induced
decidual response (default-mg/kg)
[0238] All publications cited in this specification are
incorporated herein by reference herein. While the invention has
been described with reference to a particularly preferred
embodiment, it will be appreciated that modifications can be made
without departing from the spirit of the invention. Such
modifications are intended to fall within the scope of the appended
claims.
* * * * *