# Patent

## Title
6-amino-1,4-dihydro-benzo[d][1,3]oxazin-2-ones and analogs useful as progesterone receptor modulators

## Abstract

Compounds having the structure of formula I are provided. In formula I, R1 is H, OH, substituted or unsubstituted C1 to C3 alkyl, C1 to C3 perfluoroalkyl, or COR6; R6 is H, substituted or unsubstituted C1 to C4 alkyl, aryl, substituted or unsubstituted C1 to C4 alkoxy, substituted or unsubstituted C1 to C3 aminoalkyl; R2 and R3 are H, substituted or unsubstituted C1 to C6 alkyl, C1 to C6 perfluoroalkyl, substituted or unsubstituted C2 to C6 alkenyl, substituted or unsubstituted C2 to C6 alkynyl, substituted or unsubstituted C3 to C6 cycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heterocyclic; or R2 and R3 are fused to form spirocyclic rings; R4 is NHR7, OR7, NHSO2R7, or OSO2R7; Q is O, S, NR8, or CR9R10; or a pharmaceutically acceptable salt, ester, or prodrug thereof. Such compounds are useful as progesterone receptor modulators and for treating progesterone receptor related conditions

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a divisional of U.S. patent application Ser. No. 10/946,476, filed Sep. 21, 2004, now U.S. Pat. No. 7,247,625, which claims the benefit of the priority of U.S. Provisional Patent Application No. 60/509,930, filed Oct. 9, 2003, now abandoned.


BACKGROUND OF THE INVENTION
The present invention relates to compounds that are useful as modulators of the progesterone receptor, their preparation and use thereof.
Intracellular receptors (IR) form a class of structurally related gene regulators known as “ligand dependent transcription factors”. 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).
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 PR 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 the protein encoded by that gene.
A compound that binds to an IR and mimics the action of the natural hormone is termed an agonist. Conversely, a compound which inhibits the effect of the hormone is an antagonist.
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, either along or 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 which can lead to an increased risk of uterine cancers. Co-administration of a PR agonist reduces/ablates that risk.
PR antagonists can also be used in contraception. In this context they can be administered alone, in combination with a PR agonist, or in combination with a partial ER antagonist such as tamoxifen.
PR antagonists can also be useful for the treatment of hormone dependent breast cancers as well as uterine and ovarian cancers. PR antagonists can also be useful for the treatment of non-malignant chronic conditions such as uterine fibroids and endometriosis.
PR antagonists can also be useful in hormone replacement therapy for post menopausal patients in combination with a partial ER antagonist such as tamoxifen.
PR antagonists, such as mifepristone and onapristone, have been shown to be effective in a model of hormone dependent prostate cancer, which can indicate their utility in the treatment of this condition in men.
What are needed are other compounds which function as PR modulators, including PR agonists or antagonists.
SUMMARY OF THE INVENTION
In one aspect, compounds of formula I, pharmaceutically acceptable salts, tautomers, and prodrugs thereof, are provided.




Compounds of formula I bind to the progesterone receptor and function as progesterone receptor modulators.
In a further aspect, compounds of formula I can be used for contraception; in the treatment of uterine fibroids, endometriosis, dysmenorrhea, breast cancer, uterine cancer, ovarian cancer, prostate cancer, and meningioma; hormone replacement therapy, including perimenopausal, menopausal, and post-menopausal hormone replacement therapy; and in the treatment of skin disorders, including acne and hirsutism.
In yet another aspect, pharmaceutical compositions containing a compound of formula I and a physiologically compatible carrier are provided.
In a further aspect, methods for preparing compounds of formula I are provided.
In another aspect, kits containing one or more compounds of formula I and a physiologically compatible carrier are provided.
Other aspects and advantages of the present invention are described further in the following detailed description of the preferred embodiments thereof.


DETAILED DESCRIPTION OF THE INVENTION
The term “tautomer” is meant to describe a compound which can exist in more than one isomeric state.
The term “skin” is meant to describe the outer covering of a mammalian form including, without limitation, the epidermis, dermis, and subcutaneous tissues. Typically, the skin can include other components such as hair follicles and sweat glands.
The term “acne” is meant to include any skin disorder where a skin pore becomes blocked and/or thereby becomes inflamed. The term acne includes, without limitation, superficial acne, including comedones, inflamed papules, superficial cysts, pustules, and deep acne, including deep inflamed modules and pus-filled cysts. Specific acne conditions can include, but are not limited to, acne vulgaris, acne comedo, papular acne, premenstrual acne, preadolescent acne, acne venenata, acne cosmetica, pomade acne, acne detergicans, acne excoriee, gram negative acne, acne rosacea, pseudofolliculitis barbae, folliculitis, perioral dermatitis, and hiddradenitis suppurativa.
The term “hirsutism” is meant to describe a skin disorder where an overgrowth of hair is observed in areas of the body which are not normally subject to excessive hair growth.
A number of skin disorders can be treated according to the methods of the present invention and include skin disorders of the hair follicles and sebaceous glands. Preferably, skin disorders such as acne and hirsutism, among others, can be treated according to the present invention.
Other skin disorders can include dry/chapped skin, seboria, psoriasis, or alopecia. The invention is also useful for treating the skin against the effects of environmental conditions.
As used herein, the terms “anti-progestational agents”, “anti-progestins” and “progesterone receptor antagonists” are understood to be synonymous. Similarly, “progestins”, “progestational agents” and “progesterone receptor agonists” are understood to refer to compounds of the same activity.
The term “room temperature” is meant to describe a temperature of about 23 to about 25° C. However, one of skill in the art would readily understand that the specific room temperature can vary depending upon the experimental conditions.
I. Compounds of the Invention
The present invention provides compounds of formula I having the structure:




wherein, R1 is H, OH, C1 to C3 alkyl, substituted C1 to C3 alkyl, C1 to C3 perfluoroalkyl, or COR6; R6 is H, C to C4 alkyl, substituted C1 to C4 alkyl, aryl, substituted aryl, C1 to C4 alkoxy, substituted C1 to C4 alkoxy, C1 to C3 aminoalkyl, or substituted C1 to C3 aminoalkyl; R2 and R3 are independently selected from among H, C1 to C6 alkyl, substituted C1 to C6 alkyl, C1 to C6 perfluoroalkyl, C2 to C6 alkenyl, substituted C2 to C6 alkenyl, C2 to C6 alkynyl, substituted C2 to C6 alkynyl, C3 to C6 cycloalkyl, substituted C3 to C6 cycloalkyl, aryl, substituted aryl, heterocyclic, and substituted heterocyclic; or R2 and R3 are fused to form (i) an optionally substituted 3 to 8 membered saturated spirocyclic ring; (ii) an optionally substituted 3 to 8 membered spirocyclic ring having in its backbone one or more carbon-carbon double bonds; or (iii) an optionally substituted 3 to 8 membered saturated spirocyclic ring having in its backbone one to three heteroatoms selected from the group consisting of O, S, and N; R4 is NHR7, OR7, NHSO2R7, OSO2R7, NCH3R7 or NCH3SO2R7; R7 is selected from among (a) an aryl ring which is optionally substituted with one to three independent substituents selected from among H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, C2 to C3 alkenyl, substituted C2 to C3 alkenyl, C2 to C3 alkynyl, substituted C2 to C3 alkynyl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C5 to C8 aryloxy, substituted C5 to C8 aryloxy, C1 to C3 thioalkoxy, substituted C1 to C3 thioalkoxy, amino, C1 to C3 aminoalkyl, substituted C1 to C3 aminoalkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, NRCCORB, 5 or 6 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms selected from among N, O, and S; and (b) a 5 or 6 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms selected from among O, S, SO, SO2 and N and optionally substituted with one to three independent substituents selected from among H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, C2 to C3 alkenyl, substituted C2 to C3 alkenyl, C2 to C3 alkynyl, substituted C2 to C3 alkynyl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C1 to C3 thioalkoxy, substituted C1 to C3 thioalkoxy, amino, C1 to C3 aminoalkyl, substituted C1 to C3 aminoalkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, NRCCORB, and 5 or 6 membered heterocyclic ring having in its backbone 1 to 3 heteroatoms selected from among N, O, and S; RB is H, C1 to C3 alkyl, substituted C1 to C3 alkyl, aryl, substituted aryl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C1 to C3 aminoalkyl, or substituted C1 to C3 aminoalkyl; RC is H, C1 to C3 alkyl, or substituted C1 to C3 alkyl; R5 is H, OH, halogen, CN, NO2, C1 to C6 alkyl, substituted C1 to C6 alkyl, C2 to C6 alkynyl, substituted C2 to C6 alkynyl, C1 to C6 alkoxy, substituted C1 to C6 alkoxy, amino, C1 to C6 aminoalkyl, or substituted C1 to C6 aminoalkyl; Q is O, S, NR8, or CR9R10; R8 is selected from among CN, C1 to C6 alkyl, substituted C1 to C6 alkyl, C3 to C9 cycloalkyl, substituted C3 to C8 cycloalkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, SO2CF3, OR11, and NR11R12; R9 and R10 are independent substituents selected from among H, C1 to C6 alkyl, substituted C1 to C6 alkyl, C3 to C8 cycloalkyl, substituted C3 to C8 cycloalkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, NO2, CN, and CO2R11; or CR9R10 comprise a six membered ring as shown by the structure below:





R11 and R12 are independently selected from among H, C1 to C6 alkyl, substituted C1 to C6 alkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, acyl, and sulfonyl; X is O, S, NRE, or CRERF; RE and RF are independently H, C1 to C4 lower alkyl, or C1 to C4 perfluoroalkyl; or a pharmaceutically acceptable salt, tautomer, or prodrug thereof.

R5 may be present at the 5-, 7-, or 8-position of the compound.
The invention further provides for compounds of formula including 6-[(3-chlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3,4-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3,5-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4,4-diethyl-6-[(3-fluoro-5-nitrophenyl)amino]-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 3-[(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]-5-fluorobenzonitrile; 6-[(2,3-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(2,5-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3,4-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3,5-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chloro-4-fluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-acetylphenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 5-[(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]-2-fluorobenzonitrile; 6-[(3-acetyl-4-fluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-bromophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-[(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]-2-fluorobenzonitrile; 6-[(2,3-dichlorophenyl)amino]-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(2,3-dichlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-bromophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 1,4,4-trimethyl-6-[(4-nitrophenyl)amino]-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chloro-4-fluorophenyl)amino]-4,4-diethyl-5-fluoro-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chlorophenyl)amino]-4-ethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chlorophenyl)amino]-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chloro-4-fluorophenyl)amino]-4-ethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chloro-4-fluorophenyl)amino]-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 5-fluoro-3-[(1-methyl-2-oxo-1,2-dihydrospiro[3,1-benzoxazine-4,1′-cyclopentan]-6-yl)amino]benzonitrile; 6-[(4-bromophenyl)amino]-4-(4-chlorophenyl)-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-bromophenyl)amino]-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-[(1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]benzonitrile; 4-[(1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]benzonitrile; 6-[(2,4-dichlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 1,4,4-trimethyl-6-(1-naphthylamino)-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-bromophenyl)amino]-1,4-dimethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3,4-dichlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(2-chloro-4-nitrophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(2-methoxyphenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 1,4,4-trimethyl-6-[(2-methylphenyl)amino]-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(2-bromophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-(4-chlorophenyl)-6-[(4-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-bromo-2-chlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-ethoxyphenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-bromophenyl)amino]-1-ethyl-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(2-ethylphenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 1,4,4-Trimethyl-6-(4-phenoxy-phenylamino)-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-(4-hydroxy-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-[(4-bromophenyl)amino]-4,4-bis(4-chlorophenyl)-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4,4-bis(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-benzyl-6-[(4-bromophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-benzyl-6-[(2-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-benzyl-6-[(3-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-bromophenyl)amino]-1-isopropyl-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-chlorophenyl)amino]-8-methoxy-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-4-ethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-4-ethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-1,4,8-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chlorophenyl)amino]-4-ethyl-4-phenyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(3-chlorophenyl)amino]-4-ethyl-1-methyl-4-phenyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-8-methoxy-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-(3-Chloro-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-(4-Fluoro-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-(4-Methoxy-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 1,4,4-Trimethyl-6-p-tolylamino-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-(4-Bromo -phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-(3,4-Difluoro-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-(3,5-Difluoro-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; 6-(2-Fluoro-3-methoxy-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one; (4,4-dimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)-2,3-dichlorobenzenesulfonate; (1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)-2,3-dichlorobenzenesulfonate; 2,3-dichloro-N-(4,4-dimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)benzenesulfonamide; 2,3-dichloro-N-(1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)benzenesulfonamide; 6-{[4-(dimethylamino)phenyl]amino}-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; 6-[(4-chlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; and 6-[(2-chlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one; or a pharmaceutically acceptable salt, tautomer, or prodrug thereof.
The invention also provides compounds of formula I including 6-[(3,4-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazine-2-thione; and 6-[(4-bromophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazine-2-thione; or a pharmaceutically acceptable salt, tautomer, or prodrug thereof.
The invention further provides compounds of formula I including 6-(3-chloro-4-fluorophenoxy)-4,4-diethyl-1-methyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one and 3-(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazin-6-yloxy)-5-fluoro-benzonitrile; or a pharmaceutically acceptable salt, tautomer, or prodrug thereof.
The compounds of this invention can contain an asymmetric carbon atom and/or one or more asymmetric centers and can thus give rise to optical isomers and diastereomers. While 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, tautomers, or prodrugs thereof.
The term “alkyl” is used herein to refer to both straight- and branched-chain saturated aliphatic hydrocarbon groups having 1 to about 10 carbon atoms, and desirably about 1 to about 8 carbon atoms. The term “alkenyl” is used herein to refer to both straight- and branched-chain alkyl groups having one or more carbon-carbon double bonds and containing about 2 to about 10 carbon atoms. Desirably, the term alkenyl refers to an alkyl group having 1 or 2 carbon-carbon double bonds and having 2 to about 6 carbon atoms. The term “alkynyl” group is used herein to refer to both straight- and branched-chain alkyl groups having one or more carbon-carbon triple bond and having 2 to about 8 carbon atoms. Desirably, the term alkynyl refers to an alkyl group having 1 or 2 carbon-carbon triple bonds and having 2 to about 6 carbon atoms. Typically, the term “lower” is used to describe alkyl and alkenyl groups which have 6 or fewer carbon atoms.
The terms “substituted alkyl”, “substituted alkenyl”, and “substituted alkynyl” refer to alkyl, alkenyl, and alkynyl groups, respectively, having one or more substituents including, without limitation, halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio which groups can be optionally substituted. These substituents can be attached to any carbon of an alkyl, alkenyl, or alkynyl group provided that the attachment constitutes a stable chemical moiety.
The term “perfluoroalkyl” as used herein refers to a substituted alkyl group as described above substituted by from one to three fluorine groups. Desirably, the perfluoroalkyl group includes CH2F, CHF2, and CF3 groups.
The term “cycloalkyl” as used herein refers to a cyclic alkyl or hydrocarbon group having 3 to about 10 carbon atoms, and desirably 3 to about 8 carbon atoms. The cycloalkyl group may be optionally substituted with one or more substituents including halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio which groups can be optionally substituted. These substituents can be attached to any carbon of the cycloalkyl group provided that the attachment constitutes a stable chemical moiety.
The term “acyl” as used herein refers to a carbonyl substituent, i.e., a C(O)(R) group where R is a straight- or branched-chain hydrocarbon group including, without limitation, alkyl, alkenyl, and alkynyl groups. Desirably, the R groups have 1 to about 8 carbon atoms, and more preferably 1 to about 6 carbon atoms. The term “substituted acyl” refers to an acyl group which is substituted with 1 or more groups. Desirably, an acyl group can be substituted with halogen, CN, OH, and NO2 groups, among others.
The term “aryl” as used herein refers to an aromatic system which can include a single ring or multiple aromatic rings fused or linked together where at least one part of the fused or linked rings forms a conjugated aromatic system. The aryl groups can include, but are not limited to, phenyl, naphthyl, biphenyl, anthryl, tetrahydronaphthyl, phenanthryl, indene, benzonaphthyl, fluorenyl, and carbazolyl.
The term “substituted aryl” refers to an aryl group which is substituted with one or more substituents including halogen, CN, OH, NO2, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio, which groups can be optionally substituted. Desirably, a substituted aryl group is substituted with 1 to about 4 substituents.
The term “heterocyclic” or “heterocycle” as used herein refers to a stable 4- to 7-membered monocyclic or multicyclic heterocyclic ring which is saturated, partially unsaturated, or wholly unsaturated. The heterocyclic ring has in its backbone carbon atoms and one or more heteroatom including nitrogen, oxygen, and sulfur atoms. Preferably, the heterocyclic ring has 1 to about 4 heteroatoms in the backbone of the ring. When the heterocyclic ring contains nitrogen or sulfur atoms in the backbone of the ring, the nitrogen or sulfur atoms can be oxidized. The term “heterocyclic” or “heterocycle” also refers to multicyclic rings in which a heterocyclic ring is fused to an aryl ring. The heterocyclic ring can be attached to the aryl ring through a heteroatom or carbon atom provided the resultant heterocyclic ring structure is chemically stable.
A variety of heterocyclic groups are known in the art and include, without limitation, oxygen-containing rings, nitrogen-containing rings, sulfur-containing rings, mixed heteroatom-containing rings, fused heteroatom containing rings, and combinations thereof. Oxygen-containing rings include, but are not limited to, furyl, tetrahydrofuranyl, pyranyl, pyronyl, and dioxinyl rings. Nitrogen-containing rings include, without limitation, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, pyridyl, piperidinyl, 2-oxopiperidinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, triazinyl, pyrrolidinyl, and azepinyl rings. Sulfur-containing rings include, without limitation, thienyl and dithiolyl rings. Mixed heteroatom containing rings include, but are not limited to, oxathiolyl, oxazolyl, thiazolyl, oxadiazolyl, oxatriazolyl, dioxazolyl, oxathiazolyl, oxathiolyl, oxazinyl, oxathiazinyl, morpholinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, oxepinyl, thiepinyl, and diazepinyl rings. Fused heteroatom-containing rings include, but are not limited to, benzofuranyl, thionapthene, indolyl, benzazolyl, purindinyl, pyranopyrrolyl, isoindazolyl, indoxazinyl, benzoxazolyl, anthranilyl, benzopyranyl, quinolinyl, isoquinolinyl, benzodiazonyl, napthylridinyl, benzothienyl, pyridopyridinyl, benzoxazinyl, xanthenyl, acridinyl, and purinyl rings.
The term “substituted heterocyclic” or “substituted heterocycle” as used herein refers to a heterocyclic group having one or more substituents including halogen, CN, OH, NO2, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio, which groups can be optionally substituted. Preferably, a substituted heterocyclic group has 1 to about 4 substituents.
The term “aroyl” as used herein refers to a carbonyl substituent bound to a phenyl or heterocyclic group. Preferably, the aroyl heterocyclic groups include 2-pyridinyl, 3-pyridinyl, 2-furanyl, 3-furanyl, 3-thiophenyl, 2-pyrimidinyl, and 4-pyrimidinyl groups. The term “substituted aroyl” refers to an aroyl group which is substituted with 1 or more groups including, without limitation, halogen, CN, OH, and NO2.
The term “thioalkoxy” as used herein is used interchangeably with the term “thioalkyl”, with both referring to an S(alkyl) group, where the point of attachment is through the sulfur-atom and the alkyl group can be optionally substituted.
The term “arylthio” as used herein refers to the S(aryl) group, where the point of attachment is through the sulfur-atom and the aryl group can be optionally substituted.
The term “alkoxy” as used herein refers to the O(alkyl) group, where the point of attachment is through the oxygen-atom and the alkyl group is optionally substituted.
The term “aryloxy” as used herein refers to the O(aryl) group, where the point of attachment is through the oxygen-atom and the aryl group is optionally substituted.
The term “alkylcarbonyl” as used herein refers to the C(O)(alkyl) group, where the point of attachment is through the carbon-atom of the carbonyl moiety and the alkyl group is optionally substituted.
The term “alkylcarboxy” as used herein refers to the C(O)O(alkyl) group, where the point of attachment is through the carbon-atom of the carboxy moiety and the alkyl group is optionally substituted.
The term “amino” as used herein refers to a NH2 group.
The term “aminoalkyl” as used herein refers to both secondary and tertiary amines where the point of attachment is through the nitrogen-atom and the alkyl groups are optionally substituted. The alkyl groups can be the same or different.
The term “halogen” refers to Cl, Br, F, and I groups.
The term “sulfonyl” as used herein refers to the S(O)2R group wherein R is any substituent that can form a bond to the sulfur atom group, provided that the bond constitutes a stable chemical moiety. Desirably, the R group is an optionally substituted alkyl group.
The compounds of the present invention can encompass tautomeric forms of the structures provided herein characterized by the bioactivity of the drawn structures. Further, the compounds of the present invention can be used in the form of salts derived from pharmaceutically or physiologically acceptable acids, bases, alkali metals and alkaline earth metals.
Physiologically acceptable acids include those derived from inorganic and organic acids. A number of inorganic acids are known in the art and include hydrochloric, hydrobromic, hydroiodic, sulfuric, nitric, and phosphoric acids, among others. Similarly, a variety of organic acids are known in the art and include, without limitation, lactic, formic, acetic, fumaric, citric, propionic, oxalic, succinic, glycolic, glucuronic, maleic, furoic, glutamic, benzoic, anthranilic, salicylic, tartaric, malonic, mallic, phenylacetic, mandelic, embonic, methanesulfonic, ethanesulfonic, panthenoic, benzenesulfonic, toluenesulfonic, stearic, sulfanilic, alginic, and galacturonic acids, among others.
Physiologically acceptable bases include those derived from inorganic and organic bases. A number of inorganic bases are known in the art and include aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc sulfate or phosphate compounds, among others. A number of organic bases are known in the art and include, without limitation, N,N-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, and procaine, among others.
Physiologically acceptable alkali salts and alkaline earth metal salts include, without limitation, sodium, potassium, calcium and magnesium salts in the form of esters, and carbamates.
These salts, as well as other compounds of the invention can be 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. In one embodiment, the prodrugs are esters. See, e.g., B. Testa and J. Caldwell, “Prodrugs Revisited: The “Ad Hoc” Approach as a Complement to Ligand Design”, Medicinal Research Reviews, 16(3):233-241, ed., John Wiley & Sons (1996).
As described herein, the compounds of formula I and/or salts, prodrugs or tautomers thereof, are delivered alone and/or in regimens as described herein and can include the delivery of other active agents.
The compounds discussed herein also encompass “metabolites” which are unique products formed by processing the compounds of the invention by the cell or patient. Preferably, metabolites are formed in vivo.
II. Methods of Preparing the Compounds of the Invention
The compounds of this invention can be prepared following the Schemes illustrated below.
As illustrated in Scheme I, compounds 4 and 5 can be prepared by employing coupling reactions. A substituted ortho-amino benzoic acid or a derivative thereof can be treated with an organometallic reagent in a nonprotic solvent to give ortho-amino carbinol 2 under an inert atmosphere such as argon or nitrogen at about −78° C. to about room temperature. A variety of ortho-amino benzoic acids can be utilized and include ethyl esters, among others. Such ortho-amino acids are substituted with a leaving group (X) such as Br, I, Cl, or a latent coupling precursor, such as an alkoxy group, which can be converted into another leaving group, such as an Otriflate (OTf) group. The organometallic reagent utilized in the coupling can be a Grignard reagent, among others. Nonprotic solvents can include ethers such as tetrahydrofuran (THF) or diethyl ether, among others. Typically, the reaction is executed at temperatures less than about room temperature and desirably at a temperature of about −78° C. to about room temperature.
Ring closure of carbinol 2 to yield benzoxazin-2-one 3 can be performed using a condensing agent in a nonprotic solvent. A variety of condensing agents is known in the art and include, without limitation, carbonyldiimidazole (CDI), phosgene, dimethylcarbonate, or diethylcarbonate. Typically, the nonprotic solvent is THF. Desirably, ring closure is performed at about room temperature to about 65° C.
Arylation of benzoxazin-2-one 3 to yield 4 is performed by coupling using a transition metal catalyst. Typically, the transition metal catalyst is a palladium or nickel complex and can contain phosphino ligands. Examples of phosphino ligands that can be used include triphenylphosphine (Ph3P), diphenylphosphinoferrocene (dppf), diphenylphosphinoethane (dppe), dibenzylideneacetone (dba), or 2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl (BINAP), among others. Alternatively, the transition metal catalyst is a palladium salt including palladium acetate. Typically, the coupling is performed by coupling a substituted nucleophilic reagent with the 6-halo-benzoxazinone 3 in a solvent, optionally in the presence of a base, to give the compound 4. The nucleophilic reagent can be an aniline or phenol, among others. Bases that can be used in the coupling include, but are not limited to, sodium t-butoxide, cesium carbonate, potassium phosphate, or cesium fluoride. Solvents can include, without limitation, benzene, dimethylformamide (DMF), isopropanol, ethanol, dimethoxyethane (DME), ether, acetone or a mixtures thereof. The coupling reaction is generally performed under an inert atmosphere such as nitrogen or argon at temperatures from about room temperature to about 120° C.
Alkylation of the 1-position to give compound 5 can be performed using an alkylating reagent, optionally in the presence of a base in a nonprotic solvent. Typically, the alkylating reagent is an alkyl iodide, among others. Suitable bases can include sodium hydride or sodium methoxide, among others, in a nonprotic solvent including, but not limited to, THF, dimethylformamide (DMF), or dimethylsulfoxide (DMSO). Typically, the alkylation is performed at temperatures from about 0° C. to about room temperature under an inert atmosphere of argon or nitrogen.




As illustrated in Scheme II, benzoxazinone 3 can also be converted to a nucleophile such as boronic acid 6. Desirably, benzoxazinone 3 can first be combined with an organometallic reagent in a nonprotic solvent, followed by quenching using an electrophile to form 6. A variety of organometallic reagents can be utilized and include n-BuLi, among others. Typically, the nonprotic solvent is THF or diethyl ether, among others. Electrophiles that can be utilized include, without limitation, borates, including, for example, trimethyl borate or triisopropyl borate.
Alkylation of 6 at the 1-position to form boronic acid 7 can be performed to using an alkylating reagent. A variety of alkylating reagents can be utilized and include, without limitation, alkyl halides such as an alkyl bromide or alkyl iodide. Alkylation of 6 is typically performed at temperatures of about −78° C. to about room temperature under an inert atmosphere of argon or nitrogen.
Using a copper mediated coupling reaction, a substituted aniline or phenol can be coupled with boronic acid 7 in the presence of a base in a nonprotic solvent at room temperature under an inert atmosphere of argon or nitrogen, to give compound 5. Typically, the base is a trialkyl amine including, without limitation, triethyl amine; the copper reagent is copper acetate, among others; and the nonprotic solvent is methylene chloride, among others.




As illustrated in Scheme III, 7 can also be oxidized to give 6-hydroxyl benzoxazinone 8, i.e., the phenolic precursor, at about room temperature. Oxidation is typically performed using an oxidizing reagent including, but not limited to, hydrogen peroxide or 3-chloroperoxybenzoic acid, in a solvent. Typically, the solvent is dichloromethane, among others. The phenolic precursor 8 can then be converted to compound 5 by coupling using either palladium or copper mediated coupling reactions as described above.




Scheme IV describes an alternative approach to preparing 6-arylamino benzoxazinone 5. In this alternative approach, intermediate benzoxazinone 11 is prepared from compound 9 following a similar procedure as described in Scheme I.
Nitration of 11 at the 6-position then affords 6-nitrobenzoxazinone 12 using a nitrating agent in a solvent. Preferably, the nitrating agent is concentrated nitric acid, among others, and the solvent is a mixture of acetic acid and concentrated sulfuric acid, among others. 6-nitrobenzoxazinone 12 can be alkylated using an alkylating reagent, as previously described, to prepare 13. The 6-nitro group of 13 can then be reduced using a reducing agent in a protic solvent to form 6-aminobenzoxazinone 14. A variety of reducing agents is known in the art and include sodium borohydride or hydrogen in the presence of a catalyst including palladium on carbon, among others. Protic solvents that can be used in the reduction step include alcohols such as methanol, among others.
6-aminobenzoxazinone 14 can then be coupled with an aryl halide or aryl boronic acid to prepare 5 using the palladium or copper mediated coupling procedures as previously described.




As illustrated in Scheme V, sulfonate 15 can be prepared by combining 6-hydroxylbenzoxazinone 8 with an aryl sulfonyl reagent in the presence of a base. Typically, the aryl sulfonyl reagent is, without limitation, an aryl sulfonyl halide or aryl sulfonyl anhydride and the base is Hünig's base, a trialkyl amine including, but not limited to, triethylamine, pyridine, or potassium hydroxide in a solvent such as methylene chloride, pyridine, water, or THF, among others.
Sulfonamide 16 can be similarly prepared using 6-aminobenzoxazinone 14 and a similar procedure as described for the preparation of sulfonate 15.




Scheme VI illustrates an alternative approach to prepare benzoxazinone 3. Accordingly, aniline 1 is protected in a solvent, optionally in the presence of a base either as a catalyst or as an acid scavenger. Typically, the protecting group is an alkoxy carbonyl protecting group including, but not limited to, allenoxy carbonyl, t-butoxy carbonyl, benzoxy carbonyl, ethoxy carbonyl, or methoxy carbonyl. Suitable solvents include, without limitation, THF and acetonitrile.
The protected aniline is then treated with an organometallic reagent, such as that utilized in the preparation of 2, to give the carbinol 17. Suitable organometallic reagents include, without limitation, organolithium or Grignard reagents.
Treatment of 17 with a base in a solvent under an inert atmosphere of nitrogen or argon at a temperature of about room temperature to about the boiling point of the solvent affords benzoxazinone 3. Typically, the base is potassium t-butoxide, n-butyl lithium, or potassium hydroxide, among others. The solvent desirably includes toluene, THF, or alcohol, among others.




Scheme VII describes one preparation of benzoxazinone 22 having two different substituents at the 4-position. Accordingly, isatoic anhydride 18 can be combined with N-, O-dimethylhydroxyl-amine hydrochloride salt (NHOMeMe·HCl) in a protic solvent at reflux under an inert atmosphere such as argon or nitrogen to form Weinreb amide 19. The protic solvents can include, without limitation, alcohols such as ethanol or isopropanol.
Treatment of amide 19 with organometallic compounds under an inert atmosphere such as argon or nitrogen at about −78° C. to about room temperature forms amino ketone 20. A variety of organometallic compounds can be utilized to form ketone 20 and include, but are not limited to, alkyllithium, alkynyllithium, and aryllithium reagents, or their Grignard counterpart in nonprotic solvents including ethers such as THF or diethyl ether, among others.
Ketone 20 can be converted into carbinol 21 by combining 20 with an organometallic reagent as described above under an inert atmosphere such as argon or nitrogen at about −78° C. to about room temperature. Alternatively, carbinol 21 can be prepared by reducing 20 using a reducing reagent in a solvent under an inert atmosphere in the temperature ranging from 0° C. to the boiling point of the solvent. A variety of reducing agents is known in the art and includes lithium aluminum hydride or sodium borohydride, among others. The solvent can include, without limitation, THF, ether, or anhydrous alcohol.
Ring closure of carbinol 21 to yield 22 can be accomplished using condensing agents in a nonprotic solvent. A variety of condensing agents is known and includes, without limitation, carbonyldiimidazole, phosgene, dimethylcarbonate, or diethylcarbonate. Temperatures of about room temperature to about 65° C. can be utilized.




Alternatively, as illustrated in Scheme VIII, amino ketone 20 can be prepared by treatment of benzonitrile 23 in a solvent with an organometallic reagent including, without limitation, organolithium reagents or Grignard reagents under an inert atmosphere such as argon or nitrogen at temperatures ranging from about −78° C. to about room temperature. Solvents that can be used include ethers such as THF or diethyl ether, among others.




Scheme IX depicts the preparation of ortho-amino ketone 26, where R2 is a low perfluoroalkyl including a trifluoromethyl group. Specifically, substituted aniline 24 can be protected with a protecting group to give protected aniline 25 under an inert atmosphere such as argon or nitrogen at temperatures ranging from about 0° C. to about 70° C. Such protecting groups can include pivaloyl chloride or di-tert-butyl pyrocarbonate, among others. Protection can be performed in a solvent such as acetonitrile, acetone, THF, methylene chloride, chloroform, water, or mixtures thereof, among others. Desirably, the mixture includes methylene chloride and water. A base such as sodium carbonate, sodium bicarbonate, or potassium carbonate, among others, is added when an acid, such as hydrochloride, is produced as a side-product.
Protected aniline 25 can then be treated with an alkyllithium reagent, including, without limitation, n-butyllithium or s-butyllithium, under an inert atmosphere such as argon or nitrogen at about −78° C. to about room temperature. A low perfluorocarboxy derivative, including, trifluoroacetyl chloride, 1-(trifluoroacetyl)-imidazole, or ethyl trifluoroacetate in a nonprotic solvent such as, but not limited to, ether or THF can then be added to give the protected ortho-amino ketone. Subsequent removal of the protecting group to give 26 can be performed using an acid in a solvent including, without limitation, methylene chloride or water at about 0° C. to about the boiling point of the solvent. Suitable acids that can be used include trifluoroacetic acid (TFA) or an aqueous hydrochloride solution, preferably a 3 N aqueous hydrochloride solution. Converting ketone 26 to the compounds of present invention can be performed using the procedures noted above.




As described in Scheme X, 6-substituted benzoxazin-2-one can be converted to the 6-substituted benzoxazin-2-thione 27. Typically, benzoxazin-2-one is treated with a sulfur reagent in a nonprotic solvent such as, but not limited to, o-xylene, chlorobenzene, or toluene under an inert atmosphere such as argon or nitrogen at reflux. Suitable sulfur reagents can include Lawesson's reagent, among others.




III. Formulations of the Invention

The compounds of formula I as described herein can be formulated in any form suitable for the desired route of delivery using a pharmaceutically effective amount of one or more of the compounds of formula I. For example, the compositions of the invention can be formulated for delivery by oral, dermal, transdermal, intrabronchial, intranasal, intravenous, intramuscular, subcutaneous, parenteral, intraperitoneal, intranasal, vaginal, rectal, sublingual, intracranial, epidural, and intratracheal routes, or by sustained release. Preferably, delivery is oral.
A pharmaceutically effective amount of the compound(s) used according to the present invention can vary depending on the specific compound(s), mode of delivery, and any other active ingredients used in the formulation. The dosing regimen can be adjusted to provide the optimal therapeutic response. Several divided doses can be delivered daily, e.g., in divided doses 2 to 4 times a day, or a single dose can be delivered. The dose can however be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation.
Preferably, the delivery can be on a daily, weekly, or monthly basis, and more preferably on a daily delivery. Daily dosages can be lowered or raised based on the periodic delivery.
The compounds of formula I can be delivered at a daily dosage of from about 0.5 to about 500 mg. Desirably, the compound can be delivered at a daily dosage of from about 1 to about 100 mg, more desirably about 2 to about 80 mg, and even more desirably about 2 to about 50 mg.
The compounds of formula I can be combined with one or more pharmaceutically acceptable carriers or excipients including, without limitation, solid and liquid carriers which are compatible with the compositions of the present invention. Such carriers can include adjuvants, syrups, elixirs, diluents, binders, lubricants, surfactants, granulating agents, disintegrating agents, emollients, and combinations thereof. Typically, the compound is present at about 25 to about 90%, by weight and more desirably between about 5% and 60% by weight of the formulation.
Adjuvants can include, without limitation, flavoring agents, coloring agents, preservatives, and supplemental antioxidants, which can include vitamin E, ascorbic acid, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA).
Elixers and/syrups can be prepared from acceptable sweeteners such as sugar, saccharine or a biological sweetener, a flavoring agent, and/or solvent. In one embodiment, a syrup can contain about 10 to about 50% of a sugar carrier. In another embodiment, the elixir can contain about 20 to about 50% of an ethanol carrier.
Diluents can include materials in which the compound can be dispersed, dissolved, or incorporated. Preferably, the diluents include water, lower monovalent alcohols, non-ionic surfactants, and low molecular weight glycols and polyols, including propylene glycol, diethylene glycol, polyethylene glycol, polypropylene glycol, glycerol, butylene glycol, 1,2,4-butanetriol, sorbitol esters, 1,2,6-hexanetriol, ethanol, isopropanol, sorbitol esters, butanediol, ether propanol, ethoxylated ethers, propoxylated ethers, oils such as corn, peanut and sesame oils, dimethylsulfoxide (DMSO), dimethylformamide (DMF), and combinations thereof. Preferably, the diluent is water.
Desirably, solutions or suspensions of these compounds of the invention as free bases or pharmacologically acceptable salts can be prepared in water and one or more of a surfactant, including hydroxypropylcellulose, among others.
Binders can include, without limitation, cellulose, methylcellulose, hydroxymethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, polyethylene glycol, starch, sugars such as sucrose, kaolin, and lactose, among others.
Lubricants can include magnesium stearate, light anhydrous silicic acid, talc and sodium lauryl sulfate, among others.
Granulating agents can include, without limitation, silicon dioxide, microcrystalline cellulose, starch, calcium carbonate, pectin, crospovidone, and polyplasdone, among others.
Disintegrating agents can include starch, carboxymethylcellulose, hydroxypropylstarch, substituted hydroxypropylcellulose, sodium bicarbonate, dicalcium phosphate, and calcium citrate, among others
Emollients can include, without limitation, stearyl alcohol, mink oil, cetyl alcohol, oleyl alcohol, isopropyl laurate, polyethylene glycol, olive oil, petroleum jelly, palmitic acid, oleic acid, and myristyl myristate.
IV. Therapeutic Regimens
The present invention provides dosing regimens utilizing the compound(s) of formula I with a physiologically acceptable carrier. The compositions of the invention can be delivered by a route such as oral, dermal, transdermal, intrabronchial, intranasal, intravenous, intramuscular, subcutaneous, parenteral, intraperitoneal, intranasal, vaginal, rectal, sublingual, intracranial, epidural, intratracheal, or by sustained release. Preferably, delivery is oral.
In one embodiment, the compounds are delivered orally by tablet, capsule, microcapsules, dispersible powder, granule, suspension, syrup, elixir, and aerosol. Desirably, when the compositions are delivered orally, delivery is by tablets and hard- or liquid-filled capsules.
In another embodiment, the compounds are delivered intravenously, intramuscularly, subcutaneously, parenterally and intraperitoneally in the form of sterile injectable solutions, suspensions, dispersions, and powders which are fluid to the extent that easy syringe ability exits. Such injectable compositions are sterile, stable under conditions of manufacture and storage, and free of the contaminating action of microorganisms such as bacteria and fungi.
Injectable formations can be prepared by combining the compound with a liquid. The liquid can be selected from among water, glycerol, ethanol, propylene glycol and polyethylene glycol, oils, and mixtures thereof, and more preferably the liquid carrier is water. In one embodiment, the oil is vegetable oil. Optionally, the liquid carrier contains a suspending agent. In another embodiment, the liquid carrier is an isotonic medium and contains 0.05 to about 5% suspending agent.
In a further embodiment, the compounds are delivered rectally in the form of a conventional suppository.
Dispersible formulations can be prepared by combining the compounds of the invention with glycerol, liquid, polyethylene glycols and mixtures thereof in oils.
In another embodiment, the compounds are delivered vaginally in the form of a conventional suppository, cream, gel, ring, or coated intrauterine device (IUD).
In yet another embodiment, the compositions are delivered intranasally or intrabronchially in the form of an aerosol.
In a further embodiment, the compounds are delivered transdermally or by sustained release through the use of a transdermal patch containing the composition and an optional carrier that is inert to the compound, is nontoxic to the skin, and allows for delivery of the compound for systemic absorption into the blood stream. Such a carrier can be a cream, ointment, paste, gel, or occlusive device. The creams and ointments can be viscous liquid or semisolid emulsions. Pastes include absorptive powders dispersed in petroleum or hydrophilic petroleum. Further, a variety of occlusive devices can be utilized to release the active reagents into the blood stream and include semi-permeable membranes covering a reservoir contain the active reagents, or a matrix containing the reactive reagents.
The use of sustained delivery devices can be desirable, in order to avoid the necessity for the patient to take medications on a daily basis. The term “sustained delivery” is used herein to refer to delaying the release of an active agent, i.e., a compound of the invention, until after placement in a delivery environment, followed by a sustained release of the agent at a later time. A number of sustained delivery devices are known in the art and include hydrogels (U.S. Pat. Nos. 5,266,325; 4,959,217; 5,292,515), osmotic pumps (U.S. Pat. Nos. 4,295,987 and 5,273,752 and European Patent No. 314,206, among others); hydrophobic membrane materials, such as ethylenemethacrylate (EMA) and ethylenevinylacetate (EVA); bioresorbable polymer systems (International Patent Publication No. WO 98/44964 and U.S. Pat. Nos. 5,756,127 and 5,854,388); and other bioresorbable implant devices composed of, for example, polyesters, polyanhydrides, or lactic acid/glycolic acid copolymers (U.S. Pat. No. 5,817,343). For use in such sustained delivery devices, the compounds of the invention can be formulated as described herein. See, U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719.
In yet another embodiment, the compounds are topically delivered using a topical vehicle including creams, pastes, gels, ointments, lotions, liquids, solutions, suspensions, or foams or can be alone delivered prior or subsequent to the topical vehicle. Preferably, the topical vehicles are anti-comedogenic.
Skin conditioning agents can be applied prior to, concurrently with, or subsequent to the compounds of the invention. Such skin conditioning agents typically include reagents that provide a conditioning effect to the skin and/or do not clog the pores of the skin. A number of skin conditioning agents are known in the art and include, without limitation, skin conditioning agents that can be applied to the skin, including water-based lotions, creams, pastes, gels, ointments or foams.
Optionally, other conventional acne-reducing compounds are included in the compositions and/or regimens of the invention. Such acne-reducing compounds can assist in the reduction of redness and/or blemishes. A large number of acne-reducing compounds are known in the art and include carotenoid agents, vitamin B sources, zinc compounds, and combinations thereof. See, U.S. Pat. No. 5,962,517.
Carotenoid agents can be included in the composition of the invention or can be alone delivered prior or subsequent to the compound or composition and include those carotenoids which exhibit antioxidant behavior. Preferably, the carotenoid agent includes beta-carotene, canthaxanthin, zeaxanthin, lycopen, lutein, crocetin, capsanthin, and vitamin A sources. The vitamin A sources can include vitamin A acetate or vitamin A palmitate. More preferably, the carotenoid agent is beta-carotene.
Vitamin B sources can also be included in the composition of the invention or can be alone delivered prior or subsequent to the composition to assist or promote the formation of amino acids and collagen. Preferably, the vitamin B source is a B6 source, which can include, without limitation, pyridoxine, pyridoxal, and pyridoxamine, and more preferably is pyridoxine.
Further, zinc compounds can be included in the composition of the present invention or can be alone delivered prior or subsequent to the composition. The zinc compound can include any zinc compound, preferably a zinc compound which promotes the reduction of inflammation, more preferably zinc ascorbic acid or zinc ascorbate, and most preferably zinc ascorbate.
Penetration enhancers, when used according to the method of the invention in treating hirsutism, can include any reagent that enhances the penetration of a compound through one or more layers of the skin and/or to the site of the skin disorder. A number of penetration enhancers are known in the art and include, but are not limited to, urea, propan-2-ol, polyoxyethylene ethers, terpenes, cis-fatty acids, including oleic acid and palmitoleic acid, acetone, laurocapram dimethyl sulfoxide, 2-pyrrolidone, oleyl alcohol, glyceryl-3-stearate, cholesterol, myristic acid isopropyl ester, propylene glycol, and combinations thereof.
The regimens of the invention can include the continuous delivery of the compounds of the invention. In another embodiment, the regimens can include the periodic discontinuation of delivery of the compounds of the invention. Such periodic discontinuation can include delivery of a placebo during the period of time where the compounds of the invention are not delivered to the patient. Alternatively, no placebo or active agent is delivered to the patient when the compounds are not being delivered to the patient.
By the term “placebo” or “inactive agent” is meant a reagent having pharmacological properties that are not relevant to the condition being treated, i.e., does not contain an active agent. Typical placebos include sugar as the primary constituent.
By the term “active agent” is meant any reagent which assists in treating a hormone-related condition.
The method of the present invention can be carried out over a cycle of 21 or more days, preferably 21 or more consecutive days, more preferably 21, 28, 30, or 31 days, and most preferably 21 or 28 days. One of skill in the art would readily be able to select and adjust the appropriate period of delivery.
The terminal portion of a cycle can be the last 1 to about 10 days of the cycle, and preferably the last 7 days of the cycle. In one embodiment, the terminal portion of the 28-day cycle can include the last 7 days of the cycle, i.e., days 22 to 28 of the 28-day cycle. The terminal portion of a cycle can include the delivery of an agent other than the compositions of the invention and is preferably a placebo. Alternatively, no agent or placebo is delivered during the terminal portion of the cycle.
The regimen can include delivering a daily dosage of the compound of formula I, which is incorporated into a single daily dosage unit. Delivery of the compounds of formula I can be prior to, simultaneous with, or subsequent to the delivery of other reagents that can be used according to the present invention.
The regimen can further include alternating delivery of the compounds of formula I alone, other reagent(s) that can be used according to the present invention, and a combination of the compound and the other reagent(s).
In one embodiment, a single daily dosage of the compound of formula I can be delivered for the entire 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a single daily dosage of the compound of formula I can be delivered for the first 21 days of a 28-day, 30-day, or 31-day cycle. A single daily dosage of the compound of formula I can also be delivered for the first 24 days of a 28-day, 30-day, or 31-day cycle.
The regimen can further include co-delivering the compounds of the invention with an estrogen. When included in the compositions of the present invention, the estrogens can include natural estrogens, synthetic estrogens, catechol estrogens, conjugated estrogens, and non-steroidal estrogens, among others, or pharmaceutically acceptable salts or esters thereof. In one embodiment, the estrogen is a natural estrogen including estrone, including the acetate, propionate, sulfate, and sulfate piperazine ester salts; estradiol, including the 3-benzoate, 17b-cypionate, 17-propionate, d-propionate, hemisuccinate, 17-heptanotate, 17-undecanoate, and 17-valerate ester salts; or estriol. In another embodiment, the estrogen is a synthetic estrogen including ethinyl estradiol. In a further embodiment, the estrogen is a conjugated estrogen including conjugated equine estrogens and sodium estrone sulfate and is available in formulations for intravenous, intramuscular, and topical administration (Wyeth). In a further embodiment, the estrogen is a catechol estrogen including 2- or 4-hydroxyestrogens. In yet another embodiment, the nonsteroidal estrogen is diethylstilbestrol. See, Chapter 50 entitled “Hormones” in Remington's Pharmaceutical Sciences, 18th Ed., Mack Publishing Company, Easton, Pa., 1990. The desired estrogen may however be selected from a variety of products commercially available. One of skill in the art would readily be able to select the estrogen, as well as dosage, that achieves the desired effect. Preferably, the estrogen is present in the formulation at about 0.01 mg to about 1.0 mg.
One regimen of the invention can include alternating delivery of the compounds of formula I alone, an estrogen alone, and a combination of the compound and the estrogen. The regimen can also include the delivery of another reagent prior to, in conjunction with, or subsequent to the compound of formula I and the estrogen.
In one embodiment, a single combined daily dosage of the compound of formula I and an estrogen can be delivered for the entire 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a single combined daily dosage of the compound of formula I and an estrogen can be delivered for the first 21 days of a 28-day, 30-day, or 31-day cycle. A single combined daily dosage of the compound of formula I and an estrogen can also be delivered for the first 24 days of a 28-day, 30-day, or 31-day cycle.
In a further embodiment, a daily dosage of the compound of formula I can be delivered by one route of delivery and a daily dosage of an estrogen can be delivered by a second route of delivery for the entire 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a daily dosage of the compound of formula I can be delivered by one route of delivery and a daily dosage of an estrogen can be delivered by a second route of delivery for the first 21 days of a 28-day, 30-day, or 31-day cycle. Further, a daily dosage of the compound of formula I can be delivered by one route of delivery and a daily dosage of an estrogen can be delivered by a second route of delivery for the first 24 days of a 28-day, 30-day, or 31-day cycle.
In another embodiment, a daily dosage of the compound of formula I can be delivered, followed by a daily dosage of an estrogen for the entire 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a daily dosage of the compound of formula I can be delivered, followed by a daily dosage of an estrogen for the first 21 days of a 28-day, 30-day, or 31-day cycle. Alternatively, a daily dosage of the compound of formula I can be delivered, followed by a daily dosage of an estrogen for the first 24 days of a 28-day, 30-day, or 31-day cycle.
In a further embodiment, the compounds of formula I are delivered with an estrogen for the first 14 to 24 days of a 28-day cycle, followed by delivery of the estrogen alone for a period of 1 to 11 days beginning on any cycle day between day 14 and 24.
In another embodiment, the compounds of formula I can be delivered for the initial 18 to 21 days of a 28-day cycle, followed by delivery of an estrogen alone for from 1 to 7 days.
In yet a further embodiment, the compounds of formula I can be delivered alone over a 28 day cycle for the first 21 days, followed by delivery of an estrogen alone from day 22 to day 24.
The compounds of the invention can also be delivered to a patient with one or more of a selective estrogen receptor modulator (SERM). Typically, the patient is a female of child-bearing age. The exact SERM utilized is dependent upon the patient being treated, condition being treated, and severity of the condition, among other factors.
A variety of SERMS are known in the art and include, without limitation, EM-800, EM-652, raloxifene hydrochloride, arzoxifene, lasofoxifene, droloxifene, tamoxifene, tamoxifene citrate, 4-hydroxytamoxifen citrate, clomiphene citrate, toremifene citrate, pipendoxifene, idoxifene, levormeloxifene, centchroman, nafoxidene, and bazedoxifene.
The effective dosage of the SERM employed can 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 SERMS are administered at a daily dosage of from about 0.2 to about 100 mg/kg of animal body weight, preferably given in divided doses two to four times a day, or in a sustained release form. Preferably, the amount of SERM utilized according to the present invention is preferably at least 0.2 mg per day, more preferably from about 0.2 mg to about 200 mg per day, and most preferably from about 0.2 mg to about 100 mg per day. Several divided doses can be administered daily or the dose can be proportionally reduced as indicated by the exigencies of the therapeutic situation.
Typically, a composition containing a pharmaceutically effective amount of a compound of formula I and a pharmaceutically effective amount of a SERM is delivered in a regimen to a female of child-bearing age. In one embodiment, the compound of formula I and SERM are delivered in a single composition. In another embodiment, the compound of formula I and the SERM are delivered separately. Desirably, the regimen includes delivering the composition daily for 1 to about 21 days, wherein said regimen is a cycle which is repeated monthly.
Another regimen of the invention can include alternating delivery of the compounds of formula I alone, a SERM alone, and a combination of the compound and the SERM. The regimen can also include the delivery of another reagent prior to, in conjunction with, or subsequent to the compound of formula I and the SERM.
In one embodiment, a single combined daily dosage of the compound of formula I and a SERM can be delivered for the entire 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a single combined daily dosage of the compound of formula I and a SERM can be delivered for the first 21 days of a 28-day, 30-day, or 31-day cycle. A single combined daily dosage of the compound of formula I and a SERM can also be delivered for the first 24 days of a 28-day, 30-day, or 31-day cycle.
In a further embodiment, a daily dosage of the compound of formula I can be delivered by one route of delivery and a daily dosage of a SERM can be delivered by a second route of delivery for the entire 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a daily dosage of the compound of formula I can be delivered by one route of delivery and a daily dosage of a SERM can be delivered by a second route of delivery for the first 21 days of a 28-day, 30-day, or 31-day cycle. Further, a daily dosage of the compound of formula I can be delivered by one route of delivery and a daily dosage of a SERM can be delivered by a second route of delivery for the first 24 days of a 28-day, 30-day, or 31-day cycle.
In another embodiment, a daily dosage of the compound of formula I can be delivered, followed by a daily dosage of a SERM for the entire 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a daily dosage of the compound of formula I can be delivered, followed by a daily dosage of a SERM for the first 21 days of a 28-day, 30-day, or 31-day cycle. Alternatively, a daily dosage of the compound of formula I can be delivered, followed by a daily dosage of a SERM for the first 24 days of a 28-day, 30-day, or 31-day cycle.
In a further embodiment, the compounds of formula I are delivered with a SERM for the first 14 to 24 days of a 28-day cycle, followed by delivery of the SERM alone for a period of 1 to 11 days beginning on any cycle day between day 14 and 24.
In another embodiment, the compounds of formula I can be delivered for the initial 18 to 21 days of a 28-day cycle, followed by delivery of a SERM alone for from 1 to 7 days.
In yet a further embodiment, the compounds of formula I can be delivered alone over a 28 day cycle for the first 21 days, followed by delivery of a SERM alone from day 22 to day 24.
Optionally, progestins can be delivered in combination with the compositions of the present invention. A number of progestins are known in the art and include, without limitation, progesterone, micronized progesterone, levonorgestrel, norgestrel, desogestrel, 3-ketodesogestrel, norethindrone, gestodene, norethindrone acetate, norgestimate, osaterone, cyproterone acetate, trimegestone, dienogest, drospirenone, nomegestrol, and (17-deacetyl)norgestimate, among others. Preferably, the progestins are levonorgestrel, gestodene or trimegestone.
Other reagents can be delivered in combination with the compositions of the present invention. Alternatively, such reagents can be alone administered prior or subsequent to the compositions of the invention. Such reagents can include drying agents including alcohols and benzoyl peroxides; vitamin C and D sources; amino acid reagents; enzyme activators; mineral oil; lanolin; propylene glycol; sodium lauryl sulfate; among others, and combinations thereof. Further, oral reagents include antibiotics; anti-inflammatory agents; herbal extracts including burdock root, yellow dock, horsetail, dandelion root, licorice root, echinacea, kelp, cayenne, sassafras, and elder flowers; xanthan gum; cytokines, androgens, and antiprogestins. Antibiotics can also be applied as in a topical vehicle. Such reagents can also include chemotherapeutic agents, cytokines, androgens, and antiprogestins, among others. Preferably, the chemotherapeutic agents are taxol or cisplatin. In addition, the compositions of the invention can be delivered in conjunction with other cancer treatments, including radiation therapy and/or surgery.
The term “enzyme activator” is meant to describe a reagent which activates fat and glucose metabolism and thereby results in the prevention of future acne occurrences. Preferably, the enzyme activator is a transition metal complex, more preferably is a group 5 or 6 transition metal complex, and most preferably a vanadium or chromium complex.
An isoflavone can alone be delivered or co-delivered with the compositions of the present invention in an amount sufficient to assist in the treatment of carcinomas. A number of isoflavones can be utilized and include, without limitation, genistein, daidzein, biochanin A, formononetin, and naturally occurring glucosides and glucoside conjugates. The amount of isoflavone sufficient to treat the carcinoma is dependent on the particular isoflavone utilized, the amount and activity of the co-delivered active agent, the size of the patient, the route of delivery, and the severity of the carcinoma. The amount of isoflavone sufficient to treat the hormone related condition is preferably at least 1 mg per day, more preferably from about 1 mg to about 1000 mg per day, and most preferably from about 50 mg to about 500 mg per day.
In addition, the compositions of the invention can be delivered in conjunction with other skin treatments, including laser surgery.
The dosage regimens can be adjusted to provide the optimal therapeutic response. For example, several divided doses of each component can be delivered daily or the dose can be proportionally increased or reduced as indicated by the exigencies of the therapeutic situation. In the descriptions herein, reference to a daily dosage unit can also include divided units which are delivered over the course of each day of the cycle contemplated.
V. Pharmaceutical Kits
The present invention provides kits or packages of pharmaceutical formulations designed for use in the regimens described herein. These kits are preferably designed for daily oral delivery over 21-day, 28-day, 30-day, or 31-day cycles, among others, and more preferably for one oral delivery per day. When the compositions are to be delivered continuously, a package or kit can include the composition in each tablet. When the compositions are to be delivered with periodic discontinuation, a package or kit can include placebos on those days when the composition is not delivered.
The kits are also preferably organized to indicate a single oral formulation or combination of oral formulations to be taken on each day of the cycle, preferably including oral tablets to be taken on each of the days specified, and more preferably one oral tablet will contain each of the combined daily dosages indicated.
In one embodiment, a kit can include a single phase of a daily dosage of the compound of formula I over a 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a kit can include a single phase of a daily dosage of the compound of formula I over the first 21 days of a 28-day, 30-day, or 31-day cycle. A kit can also include a single phase of a daily dosage of the compound of formula I over the first 28 days of a 30-day or 31-day cycle.
Additional components may be co-administered with the compound of formula I and include progestational agents, estrogens, and selective estrogen receptor modulators.
In a further embodiment, a kit can include a single combined phase of a daily dosage of the compound of formula I and a progestational agent over a 21-day, 28-day, 30-day, or 31-day cycle. Alternatively, a kit can include a single combined phase of a daily dosage of the compound of formula I and a progestational agent over the first 21 days of a 28-day, 30-day, or 31-day cycle. A kit can also include a single combined phase of a daily dosage of the compound of formula I and a progestational agent over the first 28 days of a 30-day or 31-day cycle.
In another embodiment, a 28-day kit can include a first phase of from 14 to 28 daily dosage units of the compound of formula I; a second phase of from 1 to 11 daily dosage units of a progestational agent; and, optionally, a third phase of an orally and pharmaceutically acceptable placebo for the remaining days of the cycle.
In yet a further embodiment, a 28-day kit can include a first phase of from 14 to 21 daily dosage units of the compound of formula I; a second phase of from 1 to 11 daily dosage units of a progestational agent; and, optionally, a third phase of an orally and pharmaceutically acceptable placebo for the remaining days of the cycle.
In another embodiment, a 28-day kit can include a first phase of from 18 to 21 daily dosage units of a compound of formula I; a second phase of from 1 to 7 daily dose units of a progestational agent; and, optionally, an orally and pharmaceutically acceptable placebo for each of the remaining 0 to 9 days in the 28-day cycle.
In yet a further embodiment, a 28-day kit can include a first phase of 21 daily dosage units of a compound of formula I; a second phase of 3 daily dosage units for days 22 to 24 of a progestational agent; and, optionally, a third phase of 4 daily units of an orally and pharmaceutically acceptable placebo for each of days 25 to 28.
In another embodiment, a 28-day kit can include a first phase of from 14 to 21 daily dosage units of a progestational agent equal in progestational activity to about 35 to about 150 μg levonorgestrel, a second phase of from 1 to 11 daily dosage units of a compound of formula I, wherein Q is O, at a daily dosage of from about 2 to 50 mg; and optionally, a third phase of an orally and pharmaceutically acceptable placebo for the remaining days of the cycle in which no antiprogestin, progestin or estrogen is administered.
In a further embodiment a 28-day kit can include a first phase of from 14 to 21 daily dosage units of a progestational agent equal in progestational activity to about 35 to about 100 μg levonorgestrel; a second phase of from 1 to 11 daily dosage units of a compound of formula I, wherein Q is O, at a daily dosage of from about 2 to 50 mg; and optionally, a third phase of an orally and pharmaceutically acceptable placebo for the remaining days of the cycle in which no antiprogestin, progestin or estrogen is administered.
Preferably, the daily dosage of each pharmaceutically active component of the regimen remains fixed in each particular phase in which it is delivered. It is further preferable that the daily dosage units described are to be delivered in the order described, with the first phase followed in order by the second and third phases. To help facilitate compliance with each regimen, it is also preferred that the kits contain the placebo described for the final days of the cycle.
A number of packages or kits are known in the art for the use in dispensing pharmaceutical agents for oral use. Preferably, the package has indicators for each day of the 28-day cycle, and more preferably is a labeled blister package, dial dispenser package, or bottle.
The compounds of the present invention encompass tautomeric forms of the structures provided herein characterized by the bioactivity of the drawn structures. Further, the compounds of the present invention can be used in the form of salts derived from pharmaceutically or physiologically acceptable acids, bases, alkali metals and alkaline earth metals.
VI. Methods of the Invention
The compounds of the present invention have a variety of uses. Desirably, the compounds of the invention are useful for treating or preventing hormone-related conditions. Specifically, the compounds of the invention are useful in treating or preventing progesterone-related conditions. In one embodiment, the compounds of the invention are useful for binding to the progesterone receptor.
Desirably, the compounds of the invention are useful in contraception. When used for such a purpose, the compounds can be administered to a mammalian subject, and preferably, a female of child-bearing age. Typically, a pharmaceutically acceptable amount of the compound is administered to the mammalian subject.
In another embodiment, the compounds of the invention are useful for treating and preventing other progesterone-related conditions and include hormone replacement therapy.
In a further embodiment, the compounds of the invention are useful for treating uterine fibroids, endometriosis, dysmenorrhea, breast cancer, uterine cancer, ovarian cancer, prostate cancer, and meningioma.
In yet another embodiment, the compounds of the invention are useful for treating skin disorder, including acne and hirsutism, among others.
The following examples are provided to illustrate the invention and do not limit the scope thereof. One skilled in the art will appreciate that although specific reagents and conditions are outlined in the following examples, modifications can be made which are meant to be encompassed by the spirit and scope of the invention.
EXAMPLES
Example 1
6-[(3-chlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
A. 4,4-Diethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one
A solution of methyl 2-amino-benzoate (15.1 grams (g), 100 millimoles (mmol)) in dry THF (200 milliliters (mL)) was treated at −78° C. under nitrogen with a solution of ethylmagnesium bromide in ether (3.0 M, 133 mL, 399 mmol). The reaction mixture was slowly warmed to ambient temperature, kept stirring for 2 hours under nitrogen and then poured into a cold 1 N aqueous hydrochloride solution (500 mL). The mixture was neutralized with aqueous 1 N sodium hydroxide solution and ethyl acetate (500 mL) was added. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine and dried (MgSO4). After removal of solvent in vacuo, the residue was taken up in anhydrous THF (200 mL) and used in the next step without further purification. To this solution was added 1,1′-carbonyldiimidazole (23.25 g, 150 mmol) under nitrogen. The reaction solution was heated at 50° C. overnight. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate (100 mL). The solution washed with 1N aqueous hydrochloride solution (2×40 mL), brine (20 mL), and dried with MgSO4. After removal of solvent in vacuo, the residue was triturated with diethyl ether to give the title compounds as white solid (14.8 g, 72%, mp 159-160° C.). 1H NMR (CDCl3): δ 8.40 (s, 1H), 7.22 (td, J=7.3, 1.6 Hz, 1H), 7.05 (td, J=7.8, 1.0 Hz, 1H), 7.02 (dd, J=7.7, 1.3 Hz, 1H), 6.80 (d, J=7.7 Hz, 1H), 1.98 (m, 4H), 0.89 (t, J=7.3 Hz, 6H); MS (EI) m/z 205; Anal. calcd for C12H15NO2: C, 70.22; H, 7.37; N, 6.82. Found: C, 69.88; H, 6.99; N, 6.31.
B. 4,4-diethyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred suspension of 4,4-diethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one (6.00 g, 29.20 mmol) in a mixture of glacial acetic acid (25 mL) and concentrated sulfuric acid (25 mL) was slowly added concentrated nitric acid (5 mL). After stirring 10 minutes, the reaction mixture was quenched with a chilled brine solution (100 mL) and ice. A solid precipitated, which was collected on a filter and washed with water. After drying in vacuo, 4,4-diethyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one was obtained as a white solid (5.02 g, 69%). 1H NMR (DMSO-d6): δ 10.85 (s, 1H), 8.20 (dd, J=8.8, 2.5 Hz, 1H), 8.06 (d, J=2.4 Hz, 1H), 7.06 (d, J=8.8 Hz, 1H), 2.14 (m, 4H), 0.79 (t, J=7.3 Hz, 6H); MS (ESI) m/z 251 ([M+H]+); MS (ESI) m/z 249 ([M−H]−); Anal. calcd for C12H14N2O4: C, 57.59; H, 5.64; N, 11.19. Found: C, 57.44; H, 5.57; N, 11.16.
C. 4,4-diethyl-1-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred solution of 4,4-diethyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one (5.00 g, 19.98 mmol) in DMF (30 mL) at 0° C. was added sodium hydride (1.20 g-60%, 29.97 mmol). After 20 minutes, methyl iodide (3.8 mL, 61.00 mmol) was added and the reaction was warmed to room temperature. After stirring for one hour, the reaction was quenched with ammonium chloride (sat.) and extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine repeatedly and dried over magnesium sulfate. Solvent was removed and the residue was triturated with ether to give 4,4-diethyl-1-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one a yellowish-white solid (3.74 g, 71%). 1H NMR (DMSO-d6): δ 8.27 (dd, J=9.0, 2.6 Hz, 1H), 8.05 (d, J=2.6 Hz, 1H), 7.34 (d, J=9.0 Hz, 1H), 3.37 (s, 3H), 2.15 (m, 4H), 0.80 (t, J=7.3 Hz, 6H); MS (ESI) m/z 265 ([M+H]+); Anal. calcd for C13H16N2O4: C, 59.08; H, 6.10; N, 10.60. Found: C, 59.62; H, 6.20; N, 10.48.
D. 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred suspension of 4,4-diethyl-1-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one (3.74 g, 14.15 mmol) and 5% palladium on carbon (0.20 g) in methanol (35 mL) at 0° C. was added sodium borohydride (10.70 g, 28.30 mmol) portionwise. After stirring 1 hr at 0° C., the reaction was quenched with ice and ammonium chloride solution (sat.) and extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine and dried over magnesium sulfate. Solvent was removed and trituration of the residue with ether gave 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one as a yellow solid (2.67 g, 81%). 1H NMR (DMSO-d6): δ 6.78 (d, J=8.6 Hz, 1H), 6.56 (m, 1H), 6.40 (d, J=2.4 Hz, 1H), 4.93 (s, 2H), 3.18 (s, 3H), 1.99 (m, 4H), 0.80 (t, J=7.3 Hz, 6H); MS (ESI) m/z 235 ([M+H]+); Anal. calcd for C13H18N2O2: C, 66.64; H, 7.74; N, 11.96. Found: C, 64.97; H, 7.67; N, 11.62.
E. 6-[(3-chlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a solution of 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (0.13 g, 0.53 mmol) and 3-chlorophenylboronic acid (0.42 g, 2.66 mmol) in methylene chloride (5 mL) was added triethylamine (0.4 mL, 2.87 mmol), and copper (II) acetate (0.19 g, 1.06 mmol) and stirred at room temperature for 2 days. The reaction was partitioned between ammonium chloride solution (sat.) and methylene chloride. The organic layer washed with sodium bicarbonate solution (sat.), dried over magnesium sulfate, and concentrated. Flash silica gel column separation with 25% ethyl acetate/hexane followed by trituration with an ether/methylene chloride/hexane mixture gave the title compound as a white solid (0.08 g, 43%). 1H NMR (DMSO-d6): δ 8.32 (s, 1H), 7.21 (t, J=8.0 Hz, 1H), 7.13 (dd, J=8.6, 2.3 Hz, 1H), 7.05 (d, J=8.7 Hz, 1H), 6.91 (m, 2H), 6.87 (dd, J=7.6, 1.4 Hz, 1H), 6.77 (d, J=7.8 Hz, 1H), 3.27 (s, 3H), 1.97 (m, 4H), 0.82 (t, J=7.3 Hz, 6H). MS (ESI) m/z 345/347 ([M+H]+); MS (ESI) m/z 343/345 ([M−H]−); Anal. calcd for C19H21ClN2O2: C, 66.18; H, 6.14; N, 8.12. Found: C, 65.70; H, 6.01; N, 7.90.
Example 2
4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
A. 4,4-Dimethyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one, which was synthesized according to the procedure depicted in example 1, using the nitration procedure described in example 1. 1H NMR (DMSO-d6): δ 11.02 (brs, 1H), 8.19 (d, J=2.4 Hz, 1H), 8.16 (m, 1H), 7.05 (dd, J=8.7, 0.5 Hz, 1H), 1.7 (s, 6H); MS (ESI) m/z 223 ([M+H]+); MS (ESI) m/z 221 ([M−H]−); Anal. calcd for C10H10N2O4: C, 54.06; H, 4.54; N, 12.61. Found: C, 52.78; H, 4.20; N, 12.23.
B. 1,4,4-Trimethyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared according to the procedure described in example 1. An off-white solid, 1H NMR (DMSO-d6): δ 8.28 (dd, J=9.0, 2.5 Hz, 1H), 8.19 (d, J=2.5 Hz, 1H), 7.34 (d, J=9.1 Hz, 1H), 3.38 (s, 3H), 1.71 (s, 6H); MS (ESI) m/z 237 ([M+H]+).
C. 6-Amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared according to the reduction procedure described in example 1. 1H NMR (DMSO-d6): δ 6.79 (d, J=8.4 Hz, 1H), 6.56 (dd, J=8.5, 2.5 Hz, 1H), 6.52 (d, J=2.4 Hz, 1H), 4.97 (s, 2H), 3.22 (s, 3H), 1.53 (s, 6H); MS (ESI) m/z 207 ([M+H]+).
D. 4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred solution of 6-amino-4-(4-chlorophenyl)-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (0.09 g, 0.30 mmol) and 1-chloro-3-iodobenzene (0.2 mL, 1.62 mmol) in THF (2 mL) was added Pd2(dba)3 (0.03 g, 0.03 mmol), BINAP (0.03 g, 0.05 mmol), sodium t-butoxide (0.07 g, 0.45 mmol), and 18-crown-6 (0.12 g, 0.45 mmol). The reaction mixture was stirred at room temperature for 2 days, quenched with ammonium chloride solution (sat.) and extracted several times with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated. Flash silica gel column separation with 20% ethyl acetate/hexane followed by trituration with hexane gave the title compound as brown solid (0.02 g, 18%). 1H NMR (DMSO-d6): δ 8.43 (s, 1H), 7.46 (dd, J=6.7, 2.0 Hz, 2H), 7.31 (dd, J=6.8, 2.2 Hz, 2H), 7.28 (m, 2H), 7.10 (d, J=10.5 Hz, 1H), 7.09 (s, 1H), 6.94 (m, 2H), 6.80 (m, 1H), 3.17 (s, 3H), 1.92 (s, 3H); MS (ESI) m/z [M+H]+(413/415); MS (ESI) m/z [M−H]−(411/413); Anal. calcd for C22H18Cl2N2O2: C, 63.93; H, 4.39; N, δ 6.78. Found: C, 62.88; H, 4.72; N, 5.67.
Example 3
6-[(3,4-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 1,2-dichloro-4-iodobenzene according to the coupling protocol described in example 2. 1H NMR (DMSO-d6): δ 8.43 (s, 1H), 7.40 (d, J=8.7 Hz, 1H), 7.13 (dd, J=8.6, 2.3 Hz, 1H), 7.06 (m, 2H), 6.93 (d, J=2.4 Hz, 1H), 6.89 (dd, J=8.8, 2.7 Hz, 1H), 3.27 (s, 3H), 1.98 (m, 4H), 0.82 (t, J=7.3 Hz, 6H); MS (ESI) m/z 379/381/383 ([M+H]+); MS (ESI) m/z 377/379/381 ([M−H]−).
Example 4
6-[(3,5-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3,5-dichlorophenylboronic acid according to the coupling protocol of example 1. 1H NMR (DMSO-d6): δ 8.55 (s, 1H), 7.16 (dd, J=8.7, 2.3 Hz, 1H), 7.09 (d, J=8.7 Hz, 1H), 6.97 (d, J=2.4 Hz, 1H), 6.84 (m, 1H), 6.82 (m, 2H), 3.28 (s, 3H), 2.00 (m, 2H), 1.92 (m, 2H), 0.82 (t, J=7.4 Hz, 6H); MS (ESI) m/z 379/381/383 ([M+H]+); MS (ESI) m/z 377/379/381 ([M−H]−); Anal. calcd for C19H20Cl2N2O2: C, 60.17; H, 5.32; N, 7.39. Found: C, 60.10; H, 5.26; N, 7.26.
Example 5
4,4-diethyl-6-[(3-fluoro-5-nitrophenyl)amino]-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 1-fluoro-3-iodo-5-nitrobenzene according to the coupling protocol of example 2. 1H NMR (DMSO-d6): δ 8.92 (s, 1H), 7.53 (t, J=1.8 Hz, 1H), 7.36 (dt, J=8.6, 2.2 Hz, 1H), 7.21 (dd, J=8.7, 2.4 Hz, 1H), 7.12 (d, J=8.7 Hz, 1H), 7.06 (d, J=2.4 Hz, 1H), 7.03 (dt, J=11.1, 2.2 Hz, 1H), 3.29 (s, 3H), 2.01 (m, 4H), 0.82 (t, J=7.3 Hz, 6H); MS (ESI) m/z 374 ([M+H]+); MS (ESI) m/z 372 ([M−H]−); Anal. calcd for C19H20FN3O4: C, 61.12; H, 5.40; N, 11.25. Found: C, 60.33; H, 5.17; N, 10.51.
Example 6
3-[(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]-5-fluorobenzonitrile
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 5-fluoro-3-cyanophenylboronic acid according to the coupling protocol of example 1. 1H NMR (DMSO-d6): δ 8.74 (s, 1H), 7.20 (dd, J=8.6, 2.3 Hz, 1H), 7.09 (m, 2H), 7.02 (m, 1H), 6.99 (d, J=2.4 Hz, 1H), 6.94 (dt, J=11.7, 2.2 Hz, 1H), 3.28 (s, 3H), 2.01 (m, 4H), 0.82 (t, J=8.3 Hz, 6H); MS (ESI) m/z 354 ([M+H]+); MS (ESI) m/z 352 ([M−H]−). Anal. calcd for C20H20FN3O2: C, 67.97; H, 5.70; N, 11.89. Found: C, 64.95; H, 5.42; N, 11.05.
Example 7
6-[(2,3-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 1,2-dichloro-3-iodobenzene according to the coupling procedure described in example 2. 1H NMR (DMSO-d6): δ 7.83 (s, 1H), 7.17 (m, 2H), 7.06 (d, J=8.7 Hz, 1H), 7.03 (m, 2H), 6.94 (dd, J=8.3, 1.4 Hz, 1H), 3.28 (s, 3H), 1.97 (m, 4H), 0.81 (t, J=7.3 Hz, 6H); MS (ESI) m/z [M+H]+(379/381/383); Anal. calcd for C19H20Cl2N2O2: C, 60.17; H, 5.32; N, 7.39. Found: C, 59.80; H, 4.96; N, 7.13.
Example 8
6-[(2,5-dichlorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 1,4-dichloro-2-iodobenzene according to the coupling procedure described in example 2. 1H NMR (DMSO-d6): δ 7.85 (s, 1H), 7.41 (d, J=8.5 Hz, 1H), 7.20 (dd, J=8.7, 2.4 Hz, 1H), 7.09 (m, 2H), 6.89 (d, J=2.4 Hz, 1H), 6.83 (dd, J=8.5, 2.4 Hz, 1H), 3.29 (s, 3H), 1.97 (m, 4H), 0.82 (t, J=7.3 Hz, 6H); MS (ESI) m/z [M+H]+(379/381/383); Anal. calcd for C19H20C12N2O2: C, 60.17; H, 5.32; N, 7.39. Found: C, 59.11; H, 5.36; N, 6.84.
Example 9
6-[(3,4-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3,4-difluorophenylboronic acid according to the coupling procedure described in example 1. 1H NMR (DMSO-d6): δ 8.24 (s, 1H), 7.28 (q, J=9.2 Hz, 1H), 7.11 (dd, J=8.7, 2.4 Hz, 1H), 7.02 (d, J=8.7 Hz, 1H), 6.90 (m, 2H), 6.74 (m, 1H), 3.26 (s, 3H), 1.97 (m, 4H), 0.81 (t, J=7.3 Hz, 6H). MS (ESI) m/z [M+H]+ (347); MS (ESI) m/z [M−H]−(345); Anal. calcd for C19H20F2N2O2: C, 65.88; H, 5.82; N, 8.09. Found: C, 65.30; H, 5.71; N, 7.79.
Example 10
6-[(3,5-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3,5-difluorophenylboronic acid according to the coupling procedure described in example 1. 1H NMR (DMSO-d6): δ 8.58 (s, 1H), 7.17 (dd, J=8.7, 2.5 Hz, 1H), 7.07 (d, J=8.7 Hz, 1H), 6.95 (d, J=2.3 Hz, 1H), 6.51 (s, 1H), 6.49 (m, 2H), 3.28 (s, 3H), 1.99 (m, 4H), 0.81 (t, J=7.4 Hz, 6H). MS (ESI) m/z 347 ([M+H]+); MS (ESI) m/z 345 ([M−H]−); Anal. calcd for C19H20F2N2O2: C, 65.88; H, 5.82; N, 8.09. Found: C, 64.99; H, 5.63; N, 7.89.
Example 11
6-[(3-chloro-4-fluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3-chloro-4-fluorophenylboronic acid according to the coupling procedure described in example 1. 1H NMR (DMSO-d6): δ 8.22 (s, 1H), 7.26 (t, J=9.1 Hz, 1H), 7.09 (dd, J=8.7, 2.3 Hz, 1H), 7.03 (m, 2H), 6.91 (m, 1H), 6.88 (d, J=2.3 Hz, 1H), 3.26 (s, 3H), 1.97 (m, 4H), 0.82 (t, J=7.2 Hz, 6H); MS (ESI) m/z 363/365 ([M+H]+); MS (ESI) m/z 361/363 ([M−H]−); Anal. calcd for C19H20ClFN2O2: C, 62.90; H, 5.56; N, 7.72. Found: C, 62.65; H, 5.53; N, 7.58.
Example 12
6-[(3-acetylphenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3-acetylphenylboronic acid according to the coupling procedure described in example 1. 1H NMR (DMSO-d6): δ 8.34 (s, 1H), 7.53 (d, J=1.8 Hz, 1H), 7.39 (m, 2H), 7.19 (dt, J=7.4, 1.9 Hz, 1H), 7.12 (dd, J=8.8, 2.5 Hz, 1H), 7.04 (d, J=8.8 Hz, 1H), 6.94 (d, J=2.5 Hz, 1H), 3.27 (s, 3H), 2.52 (s, 3H), 1.99 (m, 4H), 0.83 (t, J=7.4 Hz, 6H); MS (ESI) m/z 353 ([M+H]+); MS (ESI) m/z 351 ([M−H]−); Anal. calcd for C21H24N2O3: C, 71.57; H, 6.86; N, 7.95. Found: C, 69.83; H, 6.78; N, 7.86.
Example 13
5-[(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]-2-fluorobenzonitrile
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3-cyano-4-fluorophenylboronic acid according to the coupling procedure described in example 1. 1H NMR (DMSO-d6): δ 8.40 (s, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.27 (m, 2H), 7.14 (dd, J=8.6, 2.3 Hz, 1H), 7.04 (d, J=8.8 Hz, 1H), 6.91 (d, J=2.5 Hz, 1H), 3.27 (s, 3H), 1.98 (m, 4H), 0.81 (t, J=7.4 Hz, 6H); MS (ESI) m/z 354 ([M+H]+); MS (ESI) m/z 352 ([M−H]−); Anal. calcd for C20H20FN3O2: C, 67.97; H, 5.70; N, 11.89. Found: C, 65.58; H, 5.65; N, 11.36.
Example 14
6-[(3-acetyl-4-fluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3-acetyl-4-fluorophenylboronic acid according to the coupling procedure described in example 1. MS (ESI) m/z 371 ([M+H]+); MS (ESI) m/z 369 ([M−H]−); High Resolution Mass Spectrometry (HRMS): calcd for C21H23FN2O3, 370.1693; found (ESI+), 371.17619; Anal. calcd for C21H23FN2O3: C, 68.09; H, 6.26; N, 7.56. Found: C, 67.36; H, 6.45; N, 7.65.
Example 15
6-[(4-bromophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 1-bromo-4-iodobenzene according to the procedure described in example 2. 1H NMR (DMSO-d6): δ 8.25 (s, 1H), 7.35 (d, J=8.8 Hz, 2H), 7.11 (dd, J=8.7, 2.3 Hz, 1H), 7.02 (d, J=8.8 Hz, 1H), 6.92 (d, J=8.9 Hz, 2H), 6.87 (d, J=2.3 Hz, 1H), 3.26 (s, 3H), 1.95 (m, 4H), 0.82 (t, J=7.3 Hz, 6H); MS (ESI) m/z 389/391 ([M+H]+); MS (ESI) m/z 387/389 ([M−H]−); Anal. calcd for C19H21BrN2O2: C: 58.62H: 5.44N: 7.20. Found: C, 58.59; H, 5.29; N, 7.03.
Example 16
4-[(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]-2-fluorobenzonitrile
Prepared from 6-amino-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 4-bromo-2-fluorobenzonitrile according to the coupling procedure described in example 2. 1H NMR (DMSO-d6): δ 9.15 (s, 1H), 7.62 (t, J=7.9 Hz, 1H), 7.24 (dd, J=8.6, 2.3 Hz, 1H), 7.11 (d, J=8.7 Hz, 1H), 7.03 (d, J=2.4 Hz, 1H), 6.76 (m, 2H), 3.29 (s, 3H), 1.99 (m, 4H), 0.81 (t, J=7.3 Hz, 6H); MS (ESI) m/z 354 ([M+H]+); MS (ESI) m/z 352 ([M−H]−); Anal. calcd for C20H20FN3O2: C, 67.97; H, 5.70; N, 11.89. Found: C, 67.09; H, 5.64; N, 11.53.
Example 17
6-[(2,3-dichlorophenyl)amino]-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 2,3-difluorophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z 337/339/340 ([M+H]+); MS (ESI) m/z 335/337/339 ([M−H]−); Anal. calcd for C16H14Cl2N2O2: C, 56.99; H, 4.18; N, 8.31. Found: C, 57.46; H, 4.44; N, 7.88.
Example 18
6-[(2,3-dichlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 2,3-dichlorophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z 351/353/355 ([M+H]+); MS (ESI) m/z 349/351/353 ([M−H]−); Anal. calcd for C17H16Cl2N2O2: C, 58.13; H, 4.59; N, 7.98. Found: C, 58.59; H, 4.61; N, 7.36.
Example 19
6-[(4-bromophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 4-bromophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z 361/363 ([M+H]+); MS (ESI) m/z 359/361 ([M−H]−); Anal. calcd for C17H17BrN2O2: C, 56.52; H, 4.74; N, 7.75. Found: C, 56.98; H, 4.34; N, 7.30.
Example 20
1,4,4-trimethyl-6-[(4-nitrophenyl)amino]-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 4-nitrophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z 328 ([M+H]+); MS (ESI) m/z 326 ([M−H]−); Anal. calcd for C17H17N3O4: C, 62.38; H, 5.23; N, 12.84. Found: C, 62.03; H, 5.15; N, 11.41.
Example 21
6-[(3-chloro-4-fluorophenyl)amino]-4,4-diethyl-5-fluoro-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
A. 4,4-diethyl-5-fluoro-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred solution of 2-amino-6-fluorobenzoic acid (15.34 g, 98.90 mmol) in THF (330 mL) at 0° C. was slowly added ethyl magnesium bromide (650 mL, 650.0 mmol) and allowed to warm to room temperature. After 3 days, the reaction was quenched with chilled ammonium chloride solution (sat.) and extracted several times with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated. The crude material was taken up in THF (300 mL) and CDI (14.97 g, 92.30 mmol) was added. The reaction was stirred overnight at room temperature and partitioned between ammonium chloride solution (sat.) and ethyl acetate. The organic layer washed with 1N HCl and dried over magnesium sulfate. Trituration with ether gave 4,4-diethyl-5-fluoro-1,4-dihydro-2H-3,1-benzoxazin-2-one as an off-white solid (9.57 g, 43%). 1H NMR (DMSO-d6): δ 10.33 (s, 1H), 7.31 (m, 1H), 6.86 (m, 1H), 6.73 (d, J=8.0 Hz, 1H), 2.08 (m, 2H), 1.90 (m, 2H), 0.80 (t, J=7.2 Hz, 6H); MS (ESI) m/z 224 ([M+H]+); MS (ESI) m/z 222 ([M−H]−); Anal. calcd for C12H14FNO2: C, 64.56; H, 6.32; N, 6.27. Found: C, 64.64; H, 6.27; N, 6.16.
B. 4,4-diethyl-5-fluoro-1-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred solution of 4,4-diethyl-5-fluoro-1,4-dihydro-2H-3,1-benzoxazin-2-one (9.55 g, 42.80 mmol), prepared according to the procedure described in example 1, in THF (140 mL) at 0° C. was added sodium hydride (3.40 g, 60%, 85.00 mmol) portionwise. After 10 minutes, methyl iodide (8.0 mL, 128 mmol) was added and the reaction was allowed to warm to room temperature. After 2.5 hours the reaction was quenched with ammonium chloride solution (sat) and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The crude material was added to a mixture of glacial acetic acid (18 mL) and concentrated sulfuric acid (18 mL). Concentrated nitric acid (5 mL) was added slowly. After 10 minutes the reaction was poured into brine (150 mL) and ice. The mixture was extracted several times with ethyl acetate, and the combined organic layers were dried over magnesium sulfate and concentrated. Flash silica gel column separation with 15-30% ethyl acetate/hexane followed by trituration with ether gave 4,4-diethyl-5-fluoro-1-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one as an off-white solid (3.0 g, 25%). 1H NMR (DMSO-d6): δ 8.24 (t, J=8.5 Hz, 1H), 7.16 (dd, J=9.3, 1.0 Hz, 1H), 3.36 (s, 3H), 2.16 (m, 2H), 1.96 (m, 2H), 0.85 (t, J=7.4 Hz, 6H); MS (ESI) m/z 283 ([M+H]+); Anal. calcd for C13H15FN2O4: C, 55.32; H, 5.36; N, 9.92. Found: C, 55.29; H, 5.30; N, 9.96.
C. 6-amino-4,4-diethyl-5-fluoro-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 4,4-diethyl-5-fluoro-1-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one according to reduction procedure described in example 1. 1H NMR (DMSO-d6): δ 6.80 (t, J=8.8 Hz, 1H), 6.68 (dd, J=8.7, 1.0 Hz, 1H), 5.00 (s, 2H), 3.20 (s, 3H), 2.08 (m, 2H), 1.86 (m, 2H), 0.83 (t, J=7.3 Hz, 6H); MS (ESI) m/z 253 ([M+H]+); Anal. calcd for C13H17FN2O2: C, 61.89; H, 6.79; N, 11.10. Found: C, 61.82; H, 6.65; N, 10.96.
D. 6-[(3-chloro-4-fluorophenyl)amino]-4,4-diethyl-5-fluoro-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4,4-diethyl-5-fluoro-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3-chloro-4-fluorophenylboronic acid according to the coupling procedure described in example 1. 1H NMR (DMSO-d6): δ 8.03 (s, 1H), 7.32 (t, J=8.8 Hz, 1H), 7.26 (t, J=9.1 Hz, 1H), 6.92 (m, 2H), 6.83 (m, 1H), 3.29 (s, 3H), 2.12 (m, 2H), 1.92 (m, 2H), 0.84 (t, J=7.2 Hz, 6H); MS (ESI) m/z 381/383 ([M+H]+); MS (ESI) m/z 379/381 ([M−H]−); Anal. calcd for C19H19ClF2N2O2: C, 59.93; H, 5.03; N, 7.36. Found: C, 59.68; H, 4.92; N, 7.14.
Example 22
6-[(3-chlorophenyl)amino]-4-ethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 1-[2-amino-5-(3-chlorophenylamino)-phenyl]-1-thiophen-2-yl-propan-1-ol according to the procedures described in example 1. 1H NMR (DMSO-d6): δ 10.24 (s, 1H), 8.30 (s, 1H), 7.53 (dd, J=5.1, 1.3 Hz, 1H), 7.19 (t, J=8.1 Hz, 1H), 7.11 (dd, J=8.6, 2.4 Hz, 1H), 7.01 (m, 2H), 6.92 (dd, J=3.6, 1.3 Hz, 1H), 6.89 (m, 2H), 6.83 (m, 1H), 6.75 (m, 1H), 2.43 (m, 2H), 0.96 (t, J=7.2 Hz, 3H); MS (ESI) m/z [M+H]+(385/387); Anal. calcd for C20H17ClN2O2S: C, 62.41; H, 4.45; N, 7.28. Found: C, 62.56; H, 4.60; N, 7.01.
Example 23
6-[(3-chlorophenyl)amino]-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-[(3-chlorophenyl)amino]-4-ethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one according to the alkylation procedure described in example 1. 1H NMR (DMSO-d6): δ 8.40 (s, 1H), 7.53 (dd, J=5.0, 1.2 Hz, 1H), 7.22 (m, 2H), 7.11 (d, J=8.8 Hz, 1H), 7.05 (d, J=2.4 Hz, 1H), 7.01 (m, 1H), 6.93 (m, 2H), 6.90 (m, 1H), 6.79 (m, 1H), 3.24 (s, 3H), 2.46 (m, 2H), 0.97 (t, J=7.2 Hz, 3H); MS (ESI) m/z 399/401 ([M+H]+); Anal. calcd for C21H19ClN2O2S: C, 63.23; H, 4.80; N, 7.02. Found: C, 62.94; H, 4.87; N, 6.83.
Example 24
6-[(3-chloro-4-fluorophenyl)amino]-4-ethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one
A. 2-amino-5-[(3-chloro-4-fluorophenyl)amino]benzonitrile
5-Fluoro-2-nitrobenzonitrile (25.90 g, 156.0 mmol) and 3-chloro-4-fluorophenylamine (45.40 g, 312.0 mmol) were combined in ethanol (500 mL) and heated to reflux. After 7 days, the reaction was cooled to room temperature and filtered. The solid washed with ethanol. The solid was added to a mixture of water (100 mL) and glacial acetic acid (100 mL) and heated to 90° C. Iron powder (17.80 g, 320 mmol) was added slowly over 10 minutes. After an additional 30 minutes, the reaction was allowed to cool to room temperature and filtered through magnesol. The filtrate was taken up in ethyl acetate and washed with sodium bicarbonate solution (sat) and the organic layer was filtered through magnesol, dried over magnesium sulfate and concentrated.
Trituration with ether/hexane gave 2-amino-5-[(3-chloro-4-fluorophenyl)amino]benzonitrile as a brown solid (5.9 g, 14%). 1H NMR (DMSO-d6): δ 7.85 (s, 1H), 7.20 (t, J=9.1 Hz, 1H), 7.15 (dd, J=8.8, 2.6 Hz, 1H), 7.08 (d, J=2.6 Hz, 1H), 6.84 (dd, J=6.3, 2.7 Hz, 1H), 6.81 (d, J=8.8 Hz, 1H), 6.76 (m, 1H), 5.80 (s, 2H); MS (ESI) m/z 262/264 ([M+H]+); MS (ESI) m/z 260/262 ([M−H]−); Anal. calcd for C13H9ClFN3: C, 59.67; H, 3.47; N, 16.06. Found: C, 59.24; H, 3.41; N, 15.87.
B. 1-{2-amino-5-[(3-chloro-4-fluorophenyl)amino]phenyl}propan-1-one
To a stirred solution of 2-amino-5-[(3-chloro-4-fluorophenyl)amino]benzonitrile (5.90 g, 22.5 mmol) in THF (120 mL) at 0° C. was slowly added ethyl magnesium bromide (80 mL, 80.0 mmol). The solution was stirred at room temperature for 2 days and quenched with 1N HCl. The resulting mixture was made alkaline with sodium bicarbonate solution (sat.) and extracted several times with ethyl acetate. The combined organic layers were dried over magnesium sulfate. Flash silica gel column separation using 20% ethyl acetate/hexane followed by trituration with ether gave 1-{2-amino-5-[(3-chloro-4-fluorophenyl)amino]phenyl}propan-1-one as a yellow solid (2.48 g, 38%). 1H NMR (DMSO-d6): δ 7.78 (s, 1H), 7.48 (d, J=2.5 Hz, 1H), 7.18 (t, J=9.1 Hz, 1H), 7.12 (dd, J=8.8, 2.6 Hz, 1H), 7.04 (s, 2H), 6.81 (m, 1H), 6.79 (d, J=8.8 Hz, 1H), 6.73 (m, 1H), 2.93 (q, J=7.2 Hz, 2H), 1.07 (t, J=7.1 Hz, 3H); MS (ESI) m/z 293/295 ([M+H]+); MS (ESI) m/z 291/293 ([M−H]−); Anal. calcd for C15H14ClFN2O: C, 61.54; H, 4.82; N, 9.57. Found: C, 61.55; H, 4.68; N, 9.47.
To a stirred solution of 1-{2-amino-5-[(3-chloro-4-fluorophenyl)amino]-phenyl}propan-1-one (2.40 g, 8.20 mmol) in THF (80 mL) was added 1.0M thienyllithium (32 mL, 32 mmol) slowly at 0° C. over 20 minutes. The reaction was stirred overnight at room temperature, quenched with ammonium chloride solution (sat.), and extracted with ethyl acetate several times. The organic layer washed with brine and dried over magnesium sulfate. The concentrated crude material was dissolved in THF (40 mL). CDI (2.66 g, 16.40 mmol) was added and the reaction was stirred overnight at room temperature. The reaction solution was partitioned between ammonium chloride solution (sat.) and ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. Flash silica gel column separation with 30% ethyl acetate/hexane followed by trituration with methylene chloride/ether gave 6-[(3-chloro-4-fluorophenyl)amino]-4-ethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one as a white solid (0.19 g, 5%). 1H NMR (DMSO-d6): δ 10.22 (s, 1H), 8.20 (s, 1H), 7.53 (dd, J=5.0, 1.3 Hz, 1H), 7.24 (t, J=9.0 Hz, 1H), 7.07 (dd, J=8.5, 2.3 Hz, 1H), 7.0 (m, 3H), 6.92 (dd, J=3.6, 1.3 Hz, 1H), 6.88 (m, 2H), 2.43 (m, 2H), 0.96 (t, J=7.1 Hz, 3H); MS (ESI) m/z 403/405 ([M+H]+); MS (ESI) m/z 401/403 ([M−H]−); HRMS: calcd for C20H16ClFN2O2S, 402.0605; found (ESI+), 403.06762; Anal. calcd for C20H16ClFN2O2S: C, 59.63; H, 4.00; N, 6.95. Found: C, 59.79; H, 4.35; N, 6.55.
Example 25
6-[(3-chloro-4-fluorophenyl)amino]-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H -3,1-benzoxazin-2-one
To a stirred solution of 6-[(3-chloro-4-fluorophenyl)amino]-4-ethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one (0.16 g, 0.39 mmol) in DMF (4 mL) was added potassium t-butoxide (0.05 g, 0.42 mmol) at room temperature. After 10 minutes, methyl iodide (0.05 mL, 0.80 mmol) was added. After 45 minutes, the reaction solution was quenched with ammonium chloride solution (sat) and extracted with ethyl acetate. The organic layer washed several times with brine and dried over magnesium sulfate. The solvent was removed to give 6-[(3-chloro-4-fluorophenyl)amino]-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one as a red solid (0.12 g, 73%). 1H NMR (DMSO-d6): δ 8.31 (s, 1H), 7.53 (m, 1H), 7.27 (t, J=9.1 Hz, 1H), 7.16 (d, J=8.7 Hz, 1H), 7.10 (d, J=8.6 Hz, 1H), 7.05 (m, 1H), 7.0 (m, 2H), 6.93 (m, 2H), 3.24 (s, 3H), 2.43 (m, 2H), 0.96 (t, J=5.9 Hz, 3H); MS (ESI) m/z 417/419 ([M+H]+); MS (ESI) m/z 415/417 ([M−H]−); Anal. calcd for C21H18ClFN2O2S: C, 60.50; H, 4.35; N, 6.72. Found: C, 60.22; H, 4.34; N, 6.63.
Example 26
5-fluoro-3-[(1-methyl-2-oxo-1,2-dihydrospiro[3,1-benzoxazine-4,1′-cyclopentan]-6-yl)amino]benzonitrile
A. spiro[4H-3,1-benzoxazine-4,1′-cyclopentan]-2(1H)-one
To a stirred solution of magnesium turnings (19.44 g, 800 mmol) and a catalytic amount of iodine in ether (500 mL) was slowly added 1,4-dibromobutane (24.0 mL, 200 mmol). The reaction was stirred until reflux ceased. The magnesium was decanted and the solution was slowly added to 2-aminobenzoic acid methyl ester (8.0 mL, 61.70 mmol) in THF (600 mL) at 0° C. The solution was stirred overnight at room temperature, quenched with ammonium chloride solution (sat.) and extracted several times with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated. The crude material was taken up in THF (150 mL) and CDI (9.17 g, 56.53 mmol) was added. The reaction mixture was stirred overnight at room temperature and partitioned between ammonium chloride solution (sat) and ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. Flash silica gel column separation with 20% ethyl acetate/hexane gave spiro[4H-3,1-benzoxazine-4,1′-cyclopentan]-2(1H)-one as a white solid (2.60 g, 27%). 1H NMR (DMSO-d6): δ 10.15 (s, 1H), 7.28 (m, 2H), 7.03 (t, J=7.6 Hz, 1H), 6.89 (dd, J=7.9, 0.8 Hz, 1H), 2.15 (m, 2H), 2.07 (m, 2H), 1.90 (m, 4H); MS (ESI) m/z 204 ([M+H]+); Anal. calcd for C12H13NO2: C, 70.92; H, 6.45; N, 6.89. Found: C, 70.84; H, 6.42; N, 6.77.
B. 6-Nitrospiro[1-methyl-4H-3,1-benzoxazine-4,1′-cyclopentan]-2(1H)-one
Prepared from spiro[4H-3,1-benzoxazine-4,1′-cyclopentan]-2(1H)-one, using the nitration protocol described in example 1. 1H NMR (DMSO-d6): δ 8.27 (dd, J=9.0, 2.6 Hz, 1H), 8.17 (d, J=2.5 Hz, 1H), 7.34 (d, J=9.0 Hz, 1H), 3.42 (s, 3H), 2.20 (m, 4H), 1.94 (m, 4H); MS (ESI) m/z 263 ([M+H]+); Anal. calcd for C13H14N2O4: C, 59.54; H, 5.38; N, 10.68. Found: C, 58.58; H, 5.18; N, 10.55.
To a stirred suspension of 6-nitrospiro[1-methyl-4H-3,1-benzoxazine-4,1′-cyclopentan]-2(1H)-one (1.90 g, 7.24 mmol) and 10% palladium on carbon (0.03 g) in methanol at 0° C. was added sodium borohydride (0.55 g, 14.49 mmol) portionwise. After stirring for 30 minutes, the reaction mixture was quenched with ammonium chloride solution (sat.) and extracted several times with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated. The crude material was taken up in methylene chloride (10 mL). 5-Fluoro-3-cyanophenylboronic acid (0.49 g, 2.97 mmol), pyridine (0.25 mL, 3.09 mmol), and copper (II) acetate (0.36 g, 1.98 mmol) were added. The reaction mixture was stirred at room temperature for 2 days and partitioned between ammonium chloride solution (sat) and methylene chloride. The organic layer washed with 1N sodium hydroxide solution twice, dried over magnesium sulfate and concentrated. Flash silica gel column separation with 30% ethyl acetate/hexane gave 5-fluoro-3-[(1-methyl-2-oxo-1,2-dihydrospiro[3,1-benzoxazine-4,1′-cyclopentan]-6-yl)amino]benzonitrile as yellow solid (0.10 g, 4%). 1H NMR (DMSO-d6): δ 8.75 (s, 1H), 7.23 (dd, J=8.7, 2.3 Hz, 1H), 7.09 (m, 4H), 6.98 (dt, J=11.6, 2.2 Hz, 1H), 3.29 (s, 3H), 2.15 (m, 2H), 1.9 (m, 2H); MS (ESI) m/z 352 ([M+H]+); MS (ESI) m/z 350 ([M−H]−); HRMS: calcd for C20H18FN3O2, 351.1383. found (ESI+), 352.14616; Anal. calcd for C20H18FN3O2: C, 68.36; H, 5.16; N, 11.96. Found: C, 67.10; H, 5.20; N, 11.56.
Example 27
6-[(4-bromophenyl)amino]-4-(4-chlorophenyl)-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
A. 4-(4-chlorophenyl)-4-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred solution of 1-(2-aminophenyl)-ethanone (8.00 g, 59.20 mmol) in THF (200 mL) at 0° C. was slowly added 4-chlorophenyl magnesium bromide (1.0 M, 178 mL, 178 mmol) in ether. The reaction was allowed to warm to room temperature. After 3 hours, the reaction mixture was quenched with ammonium chloride solution (sat.) and extracted several times with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated. The crude solid was taken up in THF (200 mL) and CDI (10.56 g, 65.10 mmol) was added. The reaction solution was stirred overnight at room temperature, poured into 1N HCl, and extracted several times with ethyl acetate. The combined organic layers were dried over magnesium sulfate and trituration with ether gave 4-(4-chlorophenyl)-4-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one as a tan solid (5.19 g, 32%). 1H NMR (DMSO-d6): δ 10.30 (s, 1H), 7.46 (m, 3H), 7.36 (td, J=7.7, 1.0 Hz, 1H), 7.25 (d, J=8.6 Hz, 2H), 7.15 (t, J=7.6 Hz, 1H), 6.92 (d, J=7.7 Hz, 1H), 1.94 (s, 3H); MS (ESI) m/z [M+H]+=(274/276); MS (ESI) m/z [M−H]−=(272/274); Anal. calcd for C15H12ClNO2: C, 65.82; H, 4.42; N, 5.12. Found: C, 65.38; H, 4.58; N, 5.00.
B. 4-(4-chlorophenyl)-4-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred suspension of 4-(4-chlorophenyl)-4-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (5.15 g, 18.80 mmol) in a mixture of glacial acetic acid (15 mL) and concentrated sulfuric acid (15 mL) was slowly added concentrated nitric acid (3 mL). After 10 minutes the reaction was poured over ice and brine (100 mL) and filtered. Trituration of the solid with ethyl acetate/ether gave 4-(4-chlorophenyl)-4-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one as a tan solid (1.92 g, 32%). 1H NMR (DMSO-d6): δ 11.04 (s, 1H), 8.29 (m, 2H), 7.47 (dd, J=6.6, 2.0 Hz, 2H), 7.32 (dd, J=6.7, 2.1 Hz, 2H), 7.13 (d, J=9.5 Hz, 1H), 2.06 (s, 3H); MS (ESI) m/z [M+H]+(319/321); MS (ESI) m/z [M−H]−(317/319); Anal. calcd for C15H11ClN2O4: C, 56.53; H, 3.48; N, 8.79. Found: C, 55.43; H, 3.37; N, 8.78.
C. 4-(4-chlorophenyl)-1,4-dimethyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 4-(4-chlorophenyl)-4-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one according to the alkylation procedure described in example 1. 1H NMR (DMSO-d6): δ 8.37 (dd, J=8.9, 2.6 Hz, 1H), 8.30 (d, J=2.5 Hz, 1H), 7.47 (m, 2H), 7.39 (d, J=9.0 Hz, 1H), 7.31 (m, 2H), 3.26 (s, 3H), 2.06 (s, 3H); MS (ESI) m/z 333 ([M+H]+); Anal. calcd for C16H13ClN2O4: C, 57.76; H, 3.94; N, 8.42. Found: C, 57.85; H, 3.89; N, 8.23.
D. 6-amino-4-(4-chlorophenyl)-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 4-(4-chlorophenyl)-1,4-dimethyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one according to the reduction procedure described in example 1. 1H NMR (DMSO-d6): δ 7.44 (dd, J=6.6, 1.9 Hz, 2H), 7.25 (dd, J=6.7, 2.0 Hz, 2H), 6.83 (d, J=9.3 Hz, 1H), 6.63 (m, 2H), 5.01 (s, 2H), 3.07 (s, 3H), 1.84 (s, 3H); MS (ESI) m/z 303/305 ([M+H]+); Anal. calcd for C16H15ClN2O2: C, 63.47; H, 4.99; N, 9.25. Found: C, 63.39; H, 5.11; N, 8.96.
E. 6-[(4-bromophenyl)amino]-4-(4-chlorophenyl)-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-4-(4-chlorophenyl)-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 4-bromophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z [M+H]+(457/459/461); MS (ESI) m/z [M−H]−(455/457/459); Anal. calcd for C22H18BrClN2O2: C, 57.73; H, 3.96; N, 6.12. Found: C, 56.88; H, 3.90; N, 5.70.
Example 28
6-[(4-bromophenyl)amino]-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared 6-amino-4-ethyl-1-methyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 4-bromophenyl boronic acid according to the coupling procedure described in Example 1. MS (ESI) m/z 443/445 ([M+H]+); MS (ESI) m/z 441/443 ([M−H]−); Anal. calcd for C21H19BrN2O2S: C, 56.89; H, 4.32; N, 6.32. Found: C, 56.65; H, 4.09; N, 6.18.
Example 29
6-[(3,4-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazine-2-thione
To a stirred solution of 6-[(3,4-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (0.10 g, 0.29 mmol) in toluene (5 mL) was added Lawesson's reagent (0.09 g, 0.23 mmol). The reaction mixture was heated to reflux overnight and partitioned between ammonium chloride solution (sat.) and ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. Flash silica gel column separation using 20% ethyl acetate/hexane followed by trituration with ether gave 6-[(3,4-difluorophenyl)amino]-4,4-diethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazine-2-thione as a white solid (0.03 g, 31%). 1H NMR (DMSO-d6): δ 8.44 (s, 1H), 7.32(q, J=9.1 Hz, 1H), 7.19(d, J=8.8 Hz, 1H), 7.12 (dd, J=8.8, 2.4 Hz, 1H), 6.97 (m, 1H), 6.88 (d, J=2.4 Hz, 1H), 6.81 (m, 1H), 3.75 (s, 3H), 1.99 (m, 4H), 0.85 (t, J=7.3 Hz, 6H); MS (ESI) m/z 363 ([M+H]+); MS (ESI) m/z 361 ([M−H]−); Anal. calcd for C19H20F2N2OS: C, 62.96; H, 5.56; N, 7.73. Found: C, 62.15; H, 5.65; N, 7.39.
Example 30
6-[(4-bromophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazine-2-thione
Prepared from 6-[(4-bromophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one using the similar procedure described in example 29. 1H NMR (DMSO-d6): δ 8.49 (s, 1H), 7.39 (d, J=8.8 Hz, 2H), 7.19 (d, J=8.8 Hz, 1H), 7.12 (dd, J=8.8, 2.3 Hz, 1H), 7.00 (m, 3H), 3.75 (s, 3H), 1.60 (s, 6H); MS (ESI) m/z 377/379 ([M+H]+); MS (ESI) m/z 375/377 ([M−H]−); Anal. calcd for C17H17BrN2OS: C, 54.12; H, 4.54; N, 7.42. Found: C, 53.61; H, 4.36; N, 7.22.
Examples 31 to 60 were prepared according to the procedures described in example 1 or example 2.
Example 31
4-[(1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)amino]benzonitrile
MS (ESI) m/z [M+H]+(308); MS (ESI) m/z [M−H]−(306); Anal. calcd for C18H17N3O2: C, 70.34; H, 5.58; N, 13.67. Found: C, 69.88; H, 5.43; N, 13.18.
Example 32
6-[(2,4-dichlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 351/353/355 ([M+H]+); MS (ESI) m/z 349/351/353 ([M−H]−); Anal. calcd for C17H16Cl2N2O2: C, 58.13; H, 4.59; N, 7.98. Found: C, 57.86; H, 4.60; N, 7.69.
Example 33
1,4,4-trimethyl-6-(1-naphthylamino)-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 333 ([M+H]+); MS (ESI) m/z 331 ([M−H]−); Anal. calcd for C21H20N2O2: C, 75.88; H, 6.06; N, 8.43. Found: C, 74.64; H, 5.94; N, 8.03.
Example 34
6-[(4-bromophenyl)amino]-1,4-dimethyl-4-thien-2-yl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 427/429 ([M−H]−).
Example 35
6-[(3,4-dichlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 351/353/355 ([M+H]+); MS (ESI) m/z 349/351/353 ([M−H]−); Anal. calcd for C17H16Cl2N2O2: C, 58.13; H, 4.59; N, 7.98. Found: C, 58.11; H, 4.68; N, 7.68.
Example 36
6-[(2-chloro-4-nitrophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 362/364 ([M+H]+); MS (ESI) m/z 360/362 ([M−H]−); Anal. calcd for C17H16ClN3O4: C, 56.44; H, 4.46; N, 11.61. Found: C, 54.92; H, 4.40; N, 10.69.
Example 37
6-[(2-methoxyphenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 313 ([M+H]+); Anal. calcd for C18H20N2O3: C, 69.21; H, 6.45; N, 8.97. Found: C, 67.78; H, 6.45; N, 8.95.
Example 38
1,4,4-trimethyl-6-[(2-methylphenyl)amino]-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 297 ([M+H]+); MS (ESI) m/z 295 ([M−H]−); Anal. calcd for C18H20N2O2: C, 72.95; H, 6.80; N, 9.45. Found: C, 72.69; H, 6.90; N, 9.41.
Example 39
6-[(2-bromophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 361/363 ([M+H]+); MS (ESI) m/z 359/361 ([M−H]−).
Example 40
4-(4-chlorophenyl)-6-[(4-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 413/415/417 ([M+H]+); MS (ESI) m/z 411/413/415 ([M−H]−); Anal. calcd for C22H18Cl2N2O2: C, 63.93; H, 4.39; N, 6.78. Found: C, 62.32; H, 4.44; N, 5.95.
Example 41
6-[(4-bromo-2-chlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 395/397/399 ([M+H]+); MS (ESI) m/z 393/395/397 ([M−H]−).
Example 42
6-[(4-ethoxyphenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z [M+H]+(327); Anal. calcd for C19H22N2O3: C, 69.92; H, 6.79; N, 8.58. Found: C, 69.37; H, 6.62; N, 8.07.
Example 43
6-[(4-bromophenyl)amino]-1-ethyl-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 375/377 ([M+H]+); MS (ESI) m/z 373/375 ([M−H]−).
Example 44
6-[(2-ethylphenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z [M+H]+(311); MS (ESI) m/z [M−H]−(309); Anal. calcd for C19H22N2O2: C, 73.52; H, 7.14; N, 9.02. Found: C, 72.96; H, 7.20; N, 8.55.
Example 45
1,4,4-Trimethyl-6-(4-phenoxy-phenylamino)-1,4-dihydro-benzo[d][1,3]oxazin-2-one
MS (ESI) m/z 389 ([M+H]+); Anal. calcd for C24H24N2O3: C, 74.21; H, 6.23; N, 7.21. Found: C, 74.01; H, 6.17; N, 6.43.
Example 46
6-(4-Hydroxy-phenylamino)-1,4,4-trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one
MS (ESI) m/z 299 ([M+H]+); MS (ESI) m/z 297 ([M−H]−); Anal. calcd for C17H18N2O3: C, 68.44; H, 6.08; N, 9.39. Found: C, 66.84; H, 5.91; N, 8.94.
Example 47
6-[(4-bromophenyl)amino]-4,4-bis(4-chlorophenyl)-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 553/555/557 ([M+H]+); MS (ESI) m/z 551/553/555 ([M−H]−); Anal. calcd for C27H19BrCl2N2O2: C, 58.51; H, 3.46; N, 5.05. Found: C, 55.00; H, 3.33; N, 4.49.
Example 48
4,4-bis(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 509/511/513 ([M+H]+); MS (ESI) m/z 507/509/511 ([M−H]−); Anal. calcd for C27H19Cl3N2O2: C, 63.61; H, 3.76; N, 5.49. Found: C, 62.98; H, 3.80; N, 5.00.
Example 49
4-benzyl-6-[(4-bromophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 437/439 ([M+H]+); MS (ESI) m/z 435/437 ([M−H]−); Anal. calcd for C23H21BrN2O2: C, 63.17; H, 4.84; N, 6.41. Found: C, 63.02; H, 4.94; N, 6.04.
Example 50
4-benzyl-6-[(2-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 393/395 ([M+H]+); MS (ESI) m/z 391/393 ([M−H]−); Anal. calcd for C23H21ClN2O2: C, 70.31; H, 5.39; N, 7.13. Found: C, 68.77; H, 5.32; N, 6.61.
Example 51
4-benzyl-6-[(3-chlorophenyl)amino]-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 393/395 ([M+H]+); MS (ESI) m/z 391/393 ([M−H]−); Anal. calcd for C23H21ClN2O2: C, 70.31; H, 5.39; N, 7.13. Found: C, 65.97; H, 5.08; N, 6.53.
Example 52
6-[(4-bromophenyl)amino]-1-isopropyl-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESD m/z 389/391 ([M+H]+); MS (ESI) m/z 387/389 ([M−H]−).
Example 53
6-[(4-chlorophenyl)amino]-8-methoxy-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 347/349 ([M+H]+); MS (ESI) m/z 345/347 ([M−H]−).
Example 54
4-(4-chlorophenyl)-8-methoxy-4-methyl-6-nitro-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 347/349 ([M−H]−); Anal. calcd for C16H13ClN2O5: C, 55.11; H, 3.76; N, 8.03. Found: C, 54.77; H, 3.87; N, 7.82.
Example 55
4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-4-ethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 411/413/415 ([M−H]−); Anal. calcd for C22H18Cl2N2O2: C, 63.93; H, 4.39; N, 6.78. Found: C, 63.24; H, 4.43; N, 6.50.
Example 56
4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-4-ethyl-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 427/429/431 ([M+H]+); MS (ESI) m/z 425/427/429 ([M−H]−).
Example 57
4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-1,4,8-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 427/429/432 ([M+H]+); MS (ESI) m/z 425/427/429 ([M−H]−); Anal. calcd for C23H20C12N2O2: C, 64.65; H, 4.72; N, 6.56. Found: C, 64.34; H, 4.74; N, 6.39.
Example 58
6-[(3-chlorophenyl)amino]-4-ethyl-4-phenyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z [M+H]+(379/381); Anal. calcd for C22H19ClN2O2: C, 69.75; H, 5.05; N, 7.39. Found: C, 69.15; H, 4.75; N, 7.11.
Example 59
6-[(3-chlorophenyl)amino]-4-ethyl-1-methyl-4-phenyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z 393/395 ([M+H]+).
Example 60
4-(4-chlorophenyl)-6-[(3-chlorophenyl)amino]-8-methoxy-1,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
MS (ESI) m/z [M+H]+(443/445/447); Anal. calcd for C23H20C12N2O3: C, 62.31; H, 4.55; N, 6.32. Found: C, 62.01; H, 4.56; N, 5.93.
Examples 61-68
The compounds in the following examples were prepared from 6-Amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and the corresponding aryl boronic acid according to the procedure described in example 1.
The compounds were purified using high performance liquid chromatography (HPLC) using a YMCTM combi pro C18 Column (5 micron, 12 nanometers (nm), 50 X 4.4 millimeters (mm)). Mobile Phase A: acetonitrile with 0.1% TFA. Mobile Phase B: Water with 0.1% TFA Conditions: 1 mL/min., 10% A to 100% A 10 minute linear gradient, 100% A for 3 minutes. UV detection was at 220 and 254 nm.













Retention
MS (ES)


Compounds
Time
m/z







6-(3-Chloro-phenylamino)-1,4,4-trimethyl-1,4-
8.43
316


dihydro-benzo[d][1,3]oxazin-2-one


6-(4-Fluoro-phenylamino)-1,4,4-trimethyl-1,4-
7.34
300


dihydro-benzo[d][1,3]oxazin-2-one


6-(4-Methoxy-phenylamino)-1,4,4-trimethyl-1,4-
7.03
312


dihydro-benzo[d][1,3]oxazin-2-one


1,4,4-Trimethyl-6-p-tolylamino-1,4-dihydro-
7.90
296


benzo[d][1,3]oxazin-2-one


6-(4-Bromo-phenylamino)-1,4,4-trimethyl-1,4-
8.33
360


dihydro-benzo[d][1,3]oxazin-2-one


6-(3,4-Difluoro-phenylamino)-1,4,4-trimethyl-
7.69
318


1,4-dihydro-benzo[d][1,3]oxazin-2-one


6-(3,5-Difluoro-phenylamino)-1,4,4-trimethyl-
7.88
318


1,4-dihydro-benzo[d][1,3]oxazin-2-one


6-(2-Fluoro-3-methoxy-phenylamino)-1,4,4-
7.19
330


trimethyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one









Example 69
6-(3-chloro-4-fluorophenoxy)-4,4-diethyl-1-methyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one
A. 4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-ylboronic acid
To a stirred solution of 4,4-diethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-ylboronic acid (4.85 g, 19.50 mmol) in THF (100 mL) at 0° C. was added sodium hydride (3.12 g-60%, 78 mmol). After 10 minutes, methyl iodide (9.7 to mL, 156.0 mmol) was added and the reaction was allowed to warm to room temperature. After one week, the reaction was quenched with ammonium chloride solution (sat.) and extracted with ethyl acetate several times. The organic layers were washed with brine, dried over magnesium sulfate and concentrated. Trituration with ether gave 4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-ylboronic acid as a yellow solid (3.97 g, 77%). 1H NMR (DMSO-d6): δ 8.03 (s, 2H), 7.78 (dd, J=8.2, 1.3 Hz, 1H), 7.60 (d, J=1.3 Hz, 1H), 7.05 (d, J=8.3 Hz, 1H), 3.28 (s, 3H), 1.99 (m, 4H), 0.78 (t, J=7.4 Hz, 6H). MS (ESI) m/z 264 ([M+H]+); MS (ESI) m/z 262 ([M−H]−); Anal. calcd for C13H18BNO4: C, 59.35; H, 6.90; N, 5.32. Found: C, 58.97; H, 6.98; N, 4.73.
B. 4,4-diethyl-6-hydroxy-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
To a stirred suspension of 4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl boronic acid (3.97 g, 15.09 mmol) in a 15% acetone and 85% water mixture solution (80 mL) was added sodium bicarbonate (3.80 g, 45.27 mmol). The resulting suspension was cooled to 0° C. and Oxone (9.09 g, 14.80 mmol) was added portionwise over 20 minutes. After 45 minutes, sodium bisulfite (4.25 g, 4.10 mmol) was added. The solution was stirred for an additional 10 minutes, diluted with water and extracted several times with ethyl acetate. The organic layers were dried over magnesium sulfate and concentrated. Trituration with ether gave 4,4-diethyl-6-hydroxy-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one as a white solid (2.45 g, 69%). 1H NMR (DMSO-d6): δ 9.32 (s, 2H), 6.90 (d, J=8.8 Hz, 1H), 6.75 (dd, J=8.7, 2.6 Hz, 1H), 6.58 (d, J=2.7 Hz, 1H), 3.22 (s, 3H), 1.99 (m, 4H), 0.79 (t, J=7.3 Hz, 6H). MS (ESI) m/z 236 ([M+H]+); MS (ESI) m/z 234 ([M−H]−); Anal. calcd for C13H17NO3: C, 66.36; H, 7.28; N, 5.95. Found: C, 66.02; H, 7.34; N, 5.74.
C. 6-(3-chloro-4-fluorophenoxy)-4,4-diethyl-1-methyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one
To a stirred solution of 4,4-diethyl-6-hydroxy-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (0.30 g, 1.28 mmol) and 3-chloro-4-fluorophenylboronic acid (0.67 g, 3.83 mmol) in methylene chloride (15 mL) was added triethylamine (0.6 mL, 4.3 mmol) and copper (II) acetate (0.46 g, 2.56 mmol). The reaction mixture was stirred at room temperature for 6 days and partitioned between ammonium chloride solution (sat.) and methylene chloride. The organic layer washed with 1N sodium hydroxide solution, dried over magnesium sulfate and concentrated to give 6-(3-chloro-4-fluorophenoxy)-4,4-diethyl-1-methyl-1,4-dihydro-benzo[d][1,3]oxazin-2-one as a yellow oil (0.04 g, 9%). 1H NMR (DMSO-d6): δ 7.45 (t, J=9.1 Hz, 1H), 7.21 (dd, J=6.2, 3.0 Hz, 1H), 7.13 (d, J=8.7 Hz, 1H), 7.08 (m, 2H), 6.99 (m, 1H), 3.29 (s, 3H), 2.01 (m, 2H), 1.88 (m, 2H), 0.79 (t, J=7.3 Hz, 6H). MS (ESI) m/z 364/366 ([M+H]+); HRMS: calcd for C19H19ClFNO3, 363.1037. found (ESI+), 364.1106.
Example 70
3-(4,4-diethyl-1-methyl-2-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazin-6-yloxy)-5-fluoro-benzonitrile
Prepared from 4,4-diethyl-6-hydroxy-1-methyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 3-cyano-5-fluorobenzeneboronic acid according to the coupling procedure described in example 69. A white solid, 1H NMR (DMSO-d6): δ 7.61 (m, 1H), 7.29 (s, 1H), 7.23 (dt, J=10.3, 2.3 Hz, 1H), 7.17 (m, 2H), 7.13 (m, 1H), 3.30 (s, 3H), 2.02 (m, 2H), 1.89 (m, 2H), 0.79 (t, J=7.2 Hz, 6H). MS (ESI) m/z 355 ([M+H]+); HRMS: calcd for C20H19FN2O3, 354.1380. found (ESI+), 355.14578.
Example 71
(4,4-dimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)-2,3-dichlorobenzenesulfonate
To a stirred solution of 6-hydroxy-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (1.00 g, 5.17 mmol), prepared according to the procedure in example 69, in methylene chloride (25 mL) at 0° C. was added triethylamine (0.9 mL, 6.45 mmol). After 10 minutes, 2,3-dichlorobenzenesulfonyl chloride (1.27 g, 5.17 mmol) was added. The solution was stirred for one hour, then partitioned between ammonium chloride solution (sat.) and methylene chloride. The organic layer was dried over magnesium sulfate and concentrated. Trituration with ether gave (4,4-dimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)-2,3-dichlorobenzenesulfonate as a white solid (1.55 g, 75%). 1H NMR (DMSO-d6): δ 10.37 (s, 1H), 8.13 (dd, J=8.2, 1.6 Hz, 1H), 7.90 (dd, J=7.9, 1.5 Hz, 1H), 7.60 (t, J=8.05 Hz, 1H), 7.02 (dd, J=8.7, 2.6 Hz, 1H), 6.96 (d, J=2.7 Hz, 1H), 6.87 (d, J=8.7 Hz, 1H), 1.48 (s, 6H). MS (ESI) m/z 400/402/404 ([M−H]−); Anal. calcd for C16H13Cl2NO5S: C, 47.78; H, 3.26; N, 3.48. Found: C, 47.14; H, 3.46; N, 3.49.
Example 72
(1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)-2,3-dichlorobenzenesulfonate
To a stirred solution of (4,4-dimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)-2,3-dichlorobenzenesulfonate (0.50 g, 1.24 mmol) in dimethylformamide (DMF) (6 mL) with potassium carbonate (0.51 g, 3.72 mmol) was added methyl iodide (0.1 mL, 1.60 mmol). After stirring overnight at room temperature, the solution was then quenched with ammonium chloride solution (sat.) and extracted several times with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated. Trituration with ether gave (1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)-2,3-dichlorobenzenesulfonate as a white solid (0.30 g, 59%): 1H NMR (DMSO-d6): δ 8.14 (dd, J=8.1, 1.5 Hz, 1H), 7.92 (dd, J=8.1, 1.5 Hz, 1H), 7.61 (t, J=8.1 Hz, 1H), 7.12 (s, 2H), 7.00 (m, 1H), 3.25 (s, 3H), 1.49 (s, 6H). MS (ESI) m/z 416/418/420 ([M+H]+); Anal. calcd for C17H15Cl2NO5S: C, 49.05; H, 3.63; N, 3.36. Found: C, 49.01; H, 3.44; N, 3.15.
Example 73
2,3-dichloro-N-(4,4-dimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)benzenesulfonamide
To a stirred solution of 6-amino-4,4-dimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (0.05 g, 0.24 mmol) in methylene chloride (2 mL) at 0° C. was added triethylamine (0.05 mL, 0.36 mmol). After 10 minutes, 2,3-dichlorobenzenesulfonyl chloride (0.06 g, 0.24 mmol) was added. The solution was stirred for 2 hours, then partitioned between ammonium chloride solution (sat.) and methylene chloride. The organic layer was dried over magnesium sulfate and concentrated. Flash silica gel column separation with 50% ethyl acetate/hexane followed by trituration with ether gave 2,3-dichloro-N-(4,4-dimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)benzenesulfonamide as a white solid (0.008 g, 8%). 1H NMR (DMSO-d6): δ 10.57 (s, 1H), 10.16 (s, 1H), 7.97 (dd, J=7.9, 1.3 Hz, 1H), 7.92 (dd, J=8.0, 1.3 Hz, 1H), 7.53 (t, J=8.0 Hz, 1H), 6.93 (m, 2H), 6.74 (d, J=8.4 Hz, 1H), 1.46 (s, 6H). MS (ESI) m/z 401/403/405 ([M+H]+); MS (ESI) m/z 399/401/403 ([M−H]−).
Example 74
2,3-dichloro-N-(1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)benzenesulfonamide
To a stirred solution of 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one (0.15 g, 0.73 mmol) in methylene chloride (5 mL) at 0° C. was added triethylamine (0.15 mL, 1.07 mmol). After 10 minutes, 2,3-dichlorobenzenesulfonyl chloride (0.27 g, 1.10 mmol) was added. The solution was stirred for 5 hours, then partitioned between ammonium chloride solution (sat.) and methylene chloride. The organic layer was dried over magnesium sulfate and concentrated. Flash silica gel column separation with 30% ethyl acetate/hexane followed by trituration with ether gave 2,3-dichloro-N-(1,4,4-trimethyl-2-oxo-1,4-dihydro-2H-3,1-benzoxazin-6-yl)benzenesulfonamide as an off-white solid (0.015 g, 5%). 1H NMR (DMSO-d6): δ 10.17 (s, 1H), 8.02 (dd, J=7.9, 1.4 Hz, 1H), 7.93 (dd, J=8.1, 1.4 Hz, 1H), 7.56 (t, J=8.0 Hz, 1H), 7.03 (m, 3H), 3.20 (s, 3H), 1.48 (s, 6H). MS (ESI) m/z 415/417/419 ([M+H]+); MS (ESI) m/z 413/415/417 ([M−H]−).
Example 75
6-{[4-(dimethylamino)phenyl]amino}-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 4-dimethylaminophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z [M+H]+(326).
Example 76
6-[(4-chlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 4-chlorophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z [M+H]+(317/319); MS (ESI) m/z [M−H]−(315/317); Anal. calcd for C17H17ClN2O2: C, 64.46; H, 5.41; N, 8.84. Found: C, 63.40; H, 5.46; N, 7.97.
Example 77
6-[(2-chlorophenyl)amino]-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one
Prepared from 6-amino-1,4,4-trimethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one and 2-chlorophenyl boronic acid according to the coupling procedure described in example 1. MS (ESI) m/z [M+H]+(317/319); MS (ESI) m/z [M−H]−(315/317); Anal. calcd for C17H17ClN2O2: C, 64.46; H, 5.41; N, 8.84. Found: C, 64.42; H, 5.29; N, 8.76.
Example 78
Pharmacology
The compounds of this invention are tested in the relevant assay as described below and will have a potency in the range of 0.01 nM to 5 μM in the in vitro assays and 0.001 to 300 mg/kg in the in vivo assays. Table 1 provides the potency of selected examples of the invention, as determined in the assays described below.







TABLE 1













 















Alk. Phos.


Compound
R
R1
R2
IC50 (nM)














1
3′-CN, 4′-F
Et
Et
10.4


2
3′-F, 5′-NO2
Et
Et
10.8


3
3′-Cl, 4′-Cl
Me
Me
100.0


4
2′-Br
Me
Me
82.8


5
3′-Cl
Et
Et
11.4


6
4′-OMe
Me
Me
100.1


7
4′-Me
Me
Me
98.4


8
3′-F, 5′-F
Et
Et
12.0


9
3′-F, 4′-Cl
Et
Et
12.0


10
3′-CN, 5′-F
Et
Et
12.5


11
2′-Cl, 3′-Cl
Me
Me
56.1


12
4′-F
Me
Me
391.5


13
4′-Br
Me
Me
54.4


14
3′-F, 4′-F
Me
Me
205.7


15
3′-CN
Me
Me
1000.0


16
4′-NO2
Me
Me
309.1


17
4′-Cl
Me
Me
203.7


18
2′-Cl
Me
Me
70.3


19
4′-CN
Me
Me
600.0


20
4′-Br
4-chloro-phenyl
Me
66.3


21
2′-Cl, 4′-Cl
Me
Me
88.9


22
4′-Br
Thien-2-yl
Et
75.3


23
4′-Br
Thien-2-yl
Me
156.6


24
4′-Br
Et
Et
45.3


25
2′-Cl, 4′-NO2
Me
Me
165.1


26
2′-Me
Me
Me
130.9


27
2′-Cl, 4′-Br
Me
Me
166.3


28
3′-Cl
4-chloro-phenyl
Me
35.8


29
2′-Et
Me
Me
300.0









A. T47D cell proliferation assay

    
    
        1. Objective:
        
            Determination of progestational and antiprogestational potency was performed by using a cell proliferation assay in T47D cells. A compound's effect on DAN synthesis in T47D cells was measured.
        
        
        2. Methods:
        
            a. Reagents:
            
                Growth medium: 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).
                Treatment medium: 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).
            
            
            b. Cell Culture:
            
                Stock T47 D cells were maintained in growth medium. For BrdU incorporation assay, cells were plated in 96-well plates (Falcon, Becton Dickinson Labware) at 10,000 cells/well in growth medium. After overnight incubation, the medium was changed to treatment medium and cells were cultured for an additional 24 hours before treatment. Stock compounds were 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 were run in full dose-response curves. The final concentration of vehicle was 0.1%. In control wells, cells receive vehicle only. Antiprogestins were tested in the presence of 0.03 nM trimegestone, the reference progestin agonist. Twenty-four hours after treatment, the medium was discarded and cells were labeled with 10 mM BrdU (Amersham Life Science, Arlington Heights, Ill.) in treatment medium for 4 hours.
            
            
            c. Cell Proliferation Assay:
            
                At the end of BrdU labeling, the medium was removed and BrdU incorporation was measured using a cell proliferation ELISA kit (#RPN 250, Amersham Life Science) according to manufacturer's instructions. Briefly, cells were fixed in ethanol containing fixative for 30 minutes, followed by incubation in a blocking buffer for 30 minutes to reduce background. Peroxidase-labeled anti-BrdU antibody was added to the wells and incubated for 60 minutes. The cells were rinsed three times with PBS and incubated with 3,3′,5,5′-tetramethylbenzidine (TMB) substrate for 10-20 minutes depending upon the potency of tested compounds. Then 25 μL of 1 M sulfuric acid was added to each well to stop color reaction and optical density read in a plate reader at 450 nm within 5 minutes.
            
            
        
        
        3. Analysis of Results:
        
            Square root-transformed data were used for analysis of variance and nonlinear dose response curve fitting for both agonist and antagonist modes. Huber weighting was used to downweight the effects of outliers. EC50 or IC50 values were calculated from the retransformed values. JMP software (SAS Institute, Inc.) was used for both one-way analysis of variance and non-linear dose response analyses in both single dose and dose response studies.
        
        
        4. Reference Compounds:
        
            Trimegestone and medroxyprogesterone acetate (MPA) were reference progestins and RU486 was the reference antiprogestin. All reference compounds were run in full dose-response curves and the EC50 or IC50 values were calculated.
        
        
    
    









TABLE 2







Estimated EC50, standard error (SE), and 95% confidence


intervals (CI) for individual studies












EC50

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

















TABLE 3







Estimated IC50, standard error, and 95% confident


interval for the antiprogestin, RU486












IC50

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










EC50: Concentration of a compound that gives half-maximal increase in BrdU incorporation with SE; IC50: Concentration of a compound that gives half-maximal decrease in 0.1 trimegestone induced BrdU incorporation with SE

B. Rat decidualization assay

    
    
        1. Objective:
        
            This procedure was used to evaluate the effect of progestins and antiprogestins on rat uterine decidualization and compare the relative potencies of various test compounds.
        
        
        2. Methods:
        
            a. Reagents
            
                Test compounds were dissolved in 100% ethanol and mixed with corn oil (vehicle). Stock solutions of the test compounds in oil (MazolaTM) were then prepared by heating ( ̃80° C.) the mixture to evaporate the ethanol. Test compounds were 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.
            
            
            b. Animals (RACUC protocol #5002)
            
                Ovariectomized mature female Sprague-Dawley rats ( ̃60-day old and 230 g) were obtained from Taconic (Taconic Farms, N.Y.) following surgery. Ovariectomy was performed at least 10 days prior to treatment to reduce circulating sex steroids. Animals were housed under 12 hr light/dark cycle and given standard rat chow and water ad libitum.
            
            
            c. Treatment
            
                Rats were weighed and randomly assigned to groups of 4 or 5 before treatment. Test compounds in 0.2 mL vehicle were administered by subcutaneous injection in the nape of the neck or by gavage using 0.5 mL. The animals were treated once daily for seven days. For testing antiprogestins, animals were given the test compounds and an EC50 dose of progesterone (5.6 mg/kg) during the first three days of treatment. Following decidual stimulation, animals continued to receive progesterone until necropsy four days later.
            
            
            d. Dosing
            
                Doses were prepared based upon mg/kg mean group body weight. In all studies, a control group receiving vehicle was included. Determination of dose-response curves was carried out using doses with half log increases (e.g. 0.1, 0.3, 1.0, 3.0 mg/kg).
            
            
            e. Decidual induction
            
                Approximately 24 hours after the third injection, decidualization was induced in one of the uterine horns by scratching the antimesometrial luminal epithelium with a blunt 21 G needle. The contralateral horn was not scratched and served as an unstimulated control. Approximately 24 hours following the final treatment, rats were sacrificed by CO2 asphyxiation and body weight measured. Uteri were removed and trimmed of fat. Decidualized (D-horn) and control (C-horn) uterine horns were weighed separately.
            
            
        
        
        3. Analysis of Results:
        
            The increase in weight of the decidualized uterine horn was calculated by D-horn/C-horn and logarithmic transformation was used to maximize normality and homogeneity of variance. The Huber M-estimator was 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.) was used for both one-way ANOVA and non-linear dose-response analyses.
        
        
        4. Reference Compounds:
        
            All progestin reference compounds were run in full dose-response curves and the EC50 for uterine wet weight was calculated.
        
        
    
    









TABLE 4







Estimated EC50, standard error (SE), and 95%


confidence intervals for individual studies











EC50

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
















TABLE 5







Estimated average EC50, standard error, and 95% confidence


intervals for dose-response curves of 3 reference compounds












EC50

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

















TABLE 6







Estimated IC50, standard error, and 95% confident


interval for the antiprogestin, RU 486












IC50

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









Concentration: Compound concentration in assay (default-mg/kg body weight)
Route of administration: Route the compound is administered to the animals
Body weight: Mean total animal body weight (default-kg)
D-horn: Wet weight of decidualized uterine horn (default-mg)
C-horn: Wet weight of control uterine horn (default-mg)
Decidual response: [(D−C)/C]×100%
Progestational activity: Compounds that induce decidualization significantly (p<0.05) compared to vehicle control are considered active
Antiprogestational activity: Compounds that decrease EC50 progesterone induced decidualization significantly (p<0.05)
EC50 for uterine weight: Concentration of compound that gives half-maximal increase in decidual response (default-mg/kg)
IC50 for uterine weight: Concentration of compound that gives half-maximal decrease in EC50 progesterone induced decidual response (default-mg/kg)
C. PRE-luciferase assay in CV-1 cells

    
    
        1. Objective:
        
            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.
        
        
        2. Methods:
        
            a. Reagents:
            
                Growth medium: 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).
                Experimental medium: 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).
            
            
            b. Cell culture, transfection, treatment, and luciferase assay
            
                Stock CV-1 cells were maintained in growth medium. Co-transfection was performed using 1.2×107 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 was carried out at 260 V and 1,000 mF in a Biorad Gene Pulser II. After electroporation, cells were resuspended in growth medium and plated in 96-well plate at 40,000 cells/well in 200 μL. Following overnight incubation, the medium was changed to experimental medium. Cells were then treated with reference or test compounds in experimental medium. Compounds were tested for antiprogestational activity in the presence of 3 nM progesterone. Twenty-four hours after treatment, the medium was discarded, cells were washed three times with D-PBS (GIBCO, BRL). Fifty mL of cell lysis buffer (Promega, Madison, Wis.) was added to each well and the plates were shaken for 15 minutes in a Titer Plate Shaker (Lab Line Instrument, Inc.). Luciferase activity was measured using luciferase reagents from Promega.
            
            
            c. Analysis of Results:
            
                Each treatment consisted of at least 4 replicates. Log transformed data were used for analysis of variance and nonlinear dose response curve fitting for both agonist and antagonist modes. Huber weighting was used to downweight the effects of outliers. EC50 or IC50 values were calculated from the retransformed values. JMP software (SAS Institute, Inc.) was used for both one-way analysis of variance and non-linear response analyses.
            
            
            d. Reference Compounds:
            
                Progesterone and trimegestone were reference progestins and RU486 was the reference antiprogestin. All reference compounds were run in full dose-response curves and the EC50 or IC50 values were calculated.
            
            
        
        
    
    









TABLE 6







Estimated EC50, 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
















TABLE 7







Estimated IC50, standard error (SE), and 95% confident interval


(CI) for the antiprogestin, RU486 from three individual studies












IC50

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










Progestational activity: Compounds that increase PRE-luciferase activity significantly (p<0.05) compared to vehicle control are considered active.

Antiprogestational activity: Compounds that decrease 3 nM progesterone induced PRE-luciferase activity significantly (p<0.05)

EC50: Concentration of a compound that gives half-maximal increase PRE-luciferase activity (default-nM) with SE.
IC50: Concentration of a compound that gives half-maximal decrease in 3 nM progesterone induced PRE-luciferase activity (default-nM) with SE.
D. T47D cell alkaline phosphatase assay

    
    
        1. Purpose:
        
            To identify progestins or antiprogestins by determining a compound's effect on alkaline phosphatase activity in T47D cells.
        
        
        2. Methods:
        
            a. Reagents:
            
                Culture medium: DMEM:F12 (1:1) (GIBCO, BRL) supplemented with 5% (v/v) charcoal stripped fetal bovine serum (not heat-inactivated), 100 U/mL penicillin, 100 mg/mL streptomycin, and 2 mM GlutaMax (GIBCO, BRL).
                Alkaline phosphatase assay buffer:
            
            
        
        
        I. 0.1 M Tris-HCl, pH 9.8, containing 0.2% Triton X-100
        II. 0.1 M Tris-HCl, pH 9.8 containing 4 mM p-nitrophenyl phosphate (Sigma).
        
            b. Cell Culture and Treatment:
            
                Frozen T47D cells were thawed in a 37° 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 μL of diluted cell suspension was added. Twenty μL 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° C. in a 5% CO2/humidified atmosphere for 24 hours. Note: For high throughput screening, one concentration of each compound was tested at 0.3 mg/mL. Based on an average molecular weight of 300 g/mol for the compounds in the library, the concentration was approximately 1 mM. Subsequently, active compounds were tested in dose response assays to determine EC50 or IC50 
            
            
            c. Alkaline Phosphatase Enzyme Assay:
            
                At the end of treatment, the medium was removed from the plate. Fifty μL of assay buffer I was added to each well. The plates were shaken in a titer plate shaker for 15 minutes. Then 150 μL of assay buffer II was added to each well. Optical density measurements were taken at 5 minute intervals for 30 minutes at a test wavelength of 405 nM.
            
            
            d. Analysis of Results:
            
                Analysis of dose-response data
                For reference and test compounds, a dose response curve was generated for dose (X-axis) vs. the rate of enzyme reaction (slope) (Y-axis). Square root-transformed data were used for analysis of variance and nonlinear dose response curve fitting for both agonist and antagonist modes. Huber weighting was used to downweight the effects of outliers. EC50 or IC50 values were calculated from the retransformed values. JMP software (SAS Institute, Inc.) was used for both one-way analysis of variance and non-linear dose response analyses in both single dose and dose response studies.
                Reference Compounds: Progesterone and trimegestone were reference progestins and RU486 was the reference antiprogestin. All reference compounds were run in full dose response curves and the EC50 or IC50 values were calculated.
            
            
        
        
    
    









TABLE 8







Estimated EC50, 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

















TABLE 9







Estimated IC50, standard error, and 95% confident


interval for the reference antiprogestin RU486


from three independent experiments












IC50

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










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.

## Claims

1. A progesterone receptor modulator having the formula:




wherein:
R1 is H, OH, C1 to C3 alkyl, substituted C1 to C3 alkyl, C1 to C3 perfluoroalkyl, or COR6;
R6 is H, C1 to C4 alkyl, substituted C1 to C4 alkyl, aryl, substituted aryl, C1 to C4 alkoxy, substituted C1 to C4 alkoxy, C1 to C3 aminoalkyl, or substituted C1 to C3 aminoalkyl;
R2 and R3 are independently selected from the group consisting of H, C1 to C6 alkyl, substituted C1 to C6 alkyl, C1 to C6 perfluoroalkyl, C2 to C6 alkenyl, substituted C2 to C6 alkenyl, C2 to C6 alkynyl, substituted C2 to C6 alkynyl, C3 to C6 cycloalkyl, substituted C3 to C6 cycloalkyl, aryl, substituted aryl, heterocyclic, and substituted heterocyclic;
or R2 and R3 are fused to form:
(i) an optionally substituted 3 to 8 membered saturated spirocyclic ring;
(ii) an optionally substituted 3 to 8 membered spirocyclic ring having in its backbone one or more carbon-carbon double bonds; or
(iii) an optionally substituted 3 to 8 membered saturated spirocyclic ring having in its backbone one to three heteroatoms selected from the group consisting of O, S, and N;
R4 is NHR7, OR7, NHSO2R7, OSO2R7, NCH3R7 or NCH3SO2R7;
R7 is selected from the group consisting of (a) and (b):
(a) an aryl ring which is optionally substituted with one to three independent substituents selected from the group consisting of H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, C2 to C3 alkenyl, substituted C2 to C3 alkenyl, C2 to C3 alkynyl, substituted C2 to C3 alkynyl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C5 to C8 aryloxy, substituted C5 to C8 aryloxy, C1 to C3 thioalkoxy, substituted C1 to C3 thioalkoxy, amino, C1 to C3 aminoalkyl, substituted C1 to C3 aminoalkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, NRCCORB, 5 or 6 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms selected from the group consisting of N, O, and S; and
(b) a 5 or 6 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms selected from the group consisting of O, S, SO, SO2 and N and optionally substituted with one to three independent substituents selected from the group consisting of H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, C2 to C3 alkenyl, substituted C2 to C3 alkenyl, C2 to C3 alkynyl, substituted C2 to C3 alkynyl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C1 to C3 thioalkoxy, substituted C1 to C3 thioalkoxy, amino, C1 to C3 aminoalkyl, substituted C1 to C3 aminoalkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, NRCCORB, and 5 or 6 membered heterocyclic ring having in its backbone 1 to 3 heteroatoms selected from the group consisting of N, O, and S;
RB is H, C1 to C3 alkyl, substituted C1 to C3 alkyl, aryl, substituted aryl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C1 to C3 aminoalkyl, or substituted C1 to C3 aminoalkyl;
RC is H, C1 to C3 alkyl, or substituted C1 to C3 alkyl;


R5 is H, OH, halogen, CN, NO2, C1 to C6 alkyl, substituted C1 to C6 alkyl, C2 to C6 alkynyl, substituted C2 to C6 alkynyl, C1 to C6 alkoxy, substituted C1 to C6 alkoxy, amino, C1 to C6 aminoalkyl, or substituted C1 to C6 aminoalkyl;
Q is O, S, NR8, or CR9R10;
R8 is selected from the group consisting of CN, C1 to C6 alkyl, substituted C1 to C6 alkyl, C3 to C8 cycloalkyl, substituted C3 to C8 cycloalkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, SO2CF3, OR11, and NR11R12;
R9 and R10 are independent substituents selected from the group consisting of H, C1 to C6 alkyl, substituted C1 to C6 alkyl, C3 to C8 cycloalkyl, substituted C3 to C8 cycloalkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, NO2, CN, and CO2R11;
or CR9R10 comprise a six membered ring as shown by the structure below:




R11 and R12 are independently selected from the group consisting of H, C1 to C6 alkyl, substituted C1 to C6 alkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, acyl, and sulfonyl;
X is S,
or a pharmaceutically acceptable salt or tautomer thereof;

wherein:
said substituted alkyl, substituted alkenyl, and substituted alkynyl is an alkyl, alkenyl, and alkynyl, respectively, comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted aryl is an aryl comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio;
said substituted cycloalkyl is a cycloalkyl comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted alkoxy is an alkoxy comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted aminoalkyl is an aminoalkyl comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted heterocyclic is a heterocycle comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio;
said substituted aryloxy is an aryloxy comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted thioalkoxy is a thioalkoxy comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio.

2. The progesterone receptor modulator according to claim 1, wherein:
R1 is selected from the group consisting of H, C1 to C3 alkyl, substituted C1 to C3 alkyl, and C1 to C3 perfluoroalkyl;
R2 and R3 are independently selected from the group consisting of H, C1 to C6 alkyl, substituted C1 to C6 alkyl, C1 to C6 perfluoroalkyl, C2 to C6 alkenyl, substituted C2 to C6 alkenyl, C2 to C6 alkynyl, substituted C2 to C6 alkynyl, C3 to C6 cycloalkyl, substituted C3 to C6 cycloalkyl, aryl, substituted aryl, heterocyclic, and substituted heterocyclic;
or R2 and R3 are fused to form the optionally substituted 3 to 8 membered saturated spirocyclic ring;
R4 is selected from the group consisting of NHR7, OR7, NHSO2R7, and SO2R7;
R7 is selected from the group consisting of (a) and (b):
(a) said aryl ring which is optionally substituted with one to three substituents independently selected from the group consisting of H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, and NRCCORB; and
(b) said 5 or 6 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms selected from the group consisting of O, S, SO, SO2 and N, wherein said heterocyclic ring is optionally substituted with one or more substituents independently selected from the group consisting of H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, and NRCCORB;
RB is selected from the group consisting of H, C1 to C3 alkyl, substituted C1 to C3 alkyl, aryl, substituted aryl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C1 to C3 aminoalkyl, and substituted C1 to C3 aminoalkyl;
RC is selected from the group consisting of H, C1 to C3 alkyl, and substituted C1 to C3 alkyl;


R5 is H, OH, halogen, CN, or NO2;
or a pharmaceutically acceptable salt or tautomer thereof.

3. The progesterone receptor modulator according to claim 1, wherein:
R4 is NHR7;
R8 is selected from the group consisting of CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, SO2CF3, and OR11;
R9 and R10 are independently selected from the group consisting of C1 to C3 alkyl, substituted C1 to C3 alkyl, NO2, CN, and CO2R11;
R11 and R12 are independently selected from the group consisting of H, C1 to C3 alkyl, substituted C1 to C3 alkyl, acyl and sulfonyl;
or a pharmaceutically acceptable salt or tautomer thereof.

4. The progesterone receptor modulator according to claim 1, wherein:
R1 is Me or CF3 
R2 and R3 are independently selected from the group consisting of C1 to C4 alkyl, substituted C1 to C4 alkyl, C2 to C4 alkenyl, substituted C2 to C4 alkenyl, C2 to C4 alkynyl, substituted C2 to C4 alkynyl, C3 to C5 cycloalkyl, substituted C3 to C5 cycloalkyl, aryl, substituted aryl, heterocyclic, and substituted heterocyclic;
or R2 and R3 are fused to form an optionally substituted 3 to 6 membered saturated spirocyclic ring;
R5 is H, OH, halogen, CN, or NO2;
Q is O or S;
or a pharmaceutically acceptable salt or tautomer thereof.

5. A pharmaceutical composition comprising a progesterone receptor modulator according to claim 1 and a physiologically compatible carrier.

6. A method for contraception comprising administering to a female of child-bearing age a progesterone receptor modulator having the formula:




wherein:
R1 is H, OH, C1 to C3 alkyl, substituted C1 to C3 alkyl, C1 to C3 perfluoroalkyl, or COR6;
R6 is H, C1 to C4 alkyl, substituted C1 to C4 alkyl, aryl, substituted aryl, C1 to C4 alkoxy, substituted C1 to C4 alkoxy, C1 to C3 aminoalkyl, or substituted C1 to C3 aminoalkyl;
R2 and R3 are independently selected from the group consisting of H, C1 to C6 alkyl, substituted C1 to C6 alkyl, C1 to C6 perfluoroalkyl, C2 to C6 alkenyl, substituted C2 to C6 alkenyl, C2 to C6 alkynyl, substituted C2 to C6 alkynyl, C3 to C6 cycloalkyl, substituted C3 to C6 cycloalkyl, aryl, substituted aryl, heterocyclic, and substituted heterocyclic;
or R2 and R3 are fused to form:
(i) an optionally substituted 3 to 8 membered saturated spirocyclic ring;
(ii) an optionally substituted 3 to 8 membered spirocyclic ring having in its backbone one or more carbon-carbon double bonds; or
(iii) an optionally substituted 3 to 8 membered saturated spirocyclic ring having in its backbone one to three heteroatoms selected from the group consisting of O, S, and N;
R4 is NHR7, OR7, NHSO2R7, OSO2R7, NCH3R7 or NCH3SO2R7;
R7 is selected from the group consisting of (a) and (b):
(a) an aryl ring which is optionally substituted with one to three independent substituents selected from the group consisting of H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, C2 to C3 alkenyl, substituted C2 to C3 alkenyl, C2 to C3 alkynyl, substituted C2 to C3 alkynyl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C5 to C8 aryloxy, substituted C5 to C8 aryloxy, C1 to C3 thioalkoxy, substituted C1 to C3 thioalkoxy, amino, C1 to C3 aminoalkyl, substituted C1 to C3 aminoalkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, NRCCORB, 5 or 6 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms selected from the group consisting of N, O, and S; and
(b) a 5 or 6 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms selected from the group consisting of O, S, SO, SO2 and N and optionally substituted with one to three independent substituents selected from the group consisting of H, halogen, OH, CN, C1 to C3 alkyl, substituted C1 to C3 alkyl, C2 to C3 alkenyl, substituted C2 to C3 alkenyl, C2 to C3 alkynyl, substituted C2 to C3 alkynyl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C1 to C3 thioalkoxy, substituted C1 to C3 thioalkoxy, amino, C1 to C3 aminoalkyl, substituted C1 to C3 aminoalkyl, NO2, C1 to C3 perfluoroalkyl, CORB, CRB═NORC, OCORB, NRCCORB, and 5 or 6 membered heterocyclic ring having in its backbone 1 to 3 heteroatoms selected from the group consisting of N, O, and S;
RB is H, C1 to C3 alkyl, substituted C1 to C3 alkyl, aryl, substituted aryl, C1 to C3 alkoxy, substituted C1 to C3 alkoxy, C1 to C3 aminoalkyl, or substituted C1 to C3 aminoalkyl;
RC is H, C1 to C3 alkyl, or substituted C1 to C3 alkyl;


R5 is H, OH, halogen, CN, NO2, C1 to C6 alkyl, substituted C1 to C6 alkyl, C2 to C6 alkynyl, substituted C2 to C6 alkynyl, C1 to C6 alkoxy, substituted C1 to C6 alkoxy, amino, C1 to C6 aminoalkyl, or substituted C1 to C6 aminoalkyl;
Q is O, S, NR8, or CR9R10;
R8 is selected from the group consisting of CN, C1 to C6 alkyl, substituted C1 to C6 alkyl, C3 to C8 cycloalkyl, substituted C3 to C8 cycloalkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, SO2CF3, OR11, and NR11R12;
R9 and R10 are independent substituents selected from the group consisting of H, C1 to C6 alkyl, substituted C1 to C6 alkyl, C3 to C8 cycloalkyl, substituted C3 to C8 cycloalkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, NO2, CN, and CO2R11;
or CR9R10 comprise a six membered ring as shown by the structure below:




R11 and R12 are independently selected from the group consisting of H, C1 to C6 alkyl, substituted C1 to C6 alkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, acyl, and sulfonyl;
X is S;

or a pharmaceutically acceptable salt or tautomer thereof;
wherein:
said substituted alkyl, substituted alkenyl, and substituted alkynyl is an alkyl, alkenyl, and alkynyl, respectively, comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted aryl is an aryl comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio;
said substituted cycloalkyl is a cycloalkyl comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted alkoxy is an alkoxy comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted aminoalkyl is an aminoalkyl comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;
said substituted heterocyclic is a heterocycle comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio;
said substituted aryloxy is an aryloxy comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio;

said substituted thioalkoxy is a thioalkoxy comprising one or more substituents selected from the group consisting of halogen, CN, OH, NO2, amino, aryl, heterocyclic, aryl, alkoxy, aryloxy, alkyloxy, alkylcarbonyl, alkylcarboxy, amino, and arylthio.

7. The method for contraception according to claim 6, which comprises administering to a female of child bearing age for 28 consecutive days:
a) a first phase of from 14 to 24 daily dosage units of a progestational agent equal in progestational activity to about 35 to about 100 μg levonorgestrel;
b) a second phase of from 1 to 11 daily dosage units, at a daily dosage of from about 2 to 50 mg of a progesterone receptor modulator of formula I; and
c) optionally, a third phase of daily dosage units of an orally and pharmaceutically acceptable placebo for the remaining days of the 28 consecutive days in which no antiprogestin, progestin or estrogen is administered; wherein the total daily dosage units of the first, second and third phases equals 28.

8. A method for treating uterine fibroids comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

9. A method for treating endometriosis comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

10. A method for treating dysmenorrhea comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

11. A method for treating breast cancer comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

12. A method for treating uterine cancer comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

13. A method for treating ovarian cancer comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

14. A method for treating prostate cancer comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

15. A method for treating meningioma comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

16. A method for treating acne comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.

17. A method for treating hirsutism comprising administering to a patient in need thereof a progesterone receptor modulator according to claim 1.