Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
  • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
  • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
  • A61P15/18—Feminine contraceptives

Definitions

• This invention relates to regimens of administering compounds which are antagonists of the progesterone receptor in combination with a progestm, an estrogen, or both
• Intracellular receptors form a class of structurally related gene regulators known as "ligand dependent transcription factors" (R M Evans, Science, 240, 889, 1988)
• the steroid receptor family is a subset of the IR family, including progesterone receptor (PR), estrogen receptor (ER), androgen receptor (AR), glucocorticoid receptor (GR), and mineralocorticoid receptor (MR)
• the natural hormone, or ligand, for the PR is the steroid progesterone, but synthetic compounds, such as medroxyprogesterone acetate or levonorgestrel, have been made which also serve as ligands Once a ligand is present in the fluid surrounding a cell, it passes through the membrane via passive diffusion, and binds to the IR to create a receptor/1 igand complex This complex binds to specific gene
• a compound that binds to an IR and mimics the action of the natural hormone is termed an agonist, whilst a compound which inhibits the effect of the hormone is an antagonist
• PR antagonists may be used in contraception In this context they may be admmistered alone (Ulmann, et al, Ann N Y Acad Sci , 261, 248, 1995), in combination with a PR agonist (Kekkonen, et al, Fertility and Sterility, 60, 610, 1993) or in combination with a partial ER antagonist such as tamoxifen (WO 96/19997 Al July 4, 1996) PR antagonists may also be useful for the treatment of hormone dependent breast cancers (Horwitz, et al, Horm. Cancer, 283, pub: Birkhaeuser, Boston, Mass., ed. Nedeckis) as well as uterine and ovarian cancers.
• hormone dependent breast cancers Horm. Cancer, 283, pub: Birkhaeuser, Boston, Mass., ed. Nedeckis
• PR antagonists may also be useful for the treatment of non-malignant chronic conditions such as fibroids (Murphy, et al, J. Clin. Endo. Metab., 76, 513, 1993) and endometriosis (Kettel, et al, Fertility and Sterility, 56, 402, 1991).
• PR antagonists may also be useful in hormone replacement therapy for post menopausal patients in combination with a partial ER antagonist such as tamoxifen (U.S. Patent No. 5,719,136).
• PR antagonists such as mifepristone and onapristone
• PR antagonists have been shown to be effective in a model of hormone dependent prostate cancer, which may indicate their utility in the treatment of this condition in men (Michna, et al, Ann. N. Y. Acad. Sci., 761, 224, 1995).
• RI F, Cl, Br, alkyl
• NH 2 R2 alkyl, alkoxy, F, Cl, NH2, CF,
• Bohm, et al include the generic structure U (WO 91/04974).
• Boar, et al described the dioxolane W as an intermediate for preparation of acetyl- cholmesterase inhibitors (WO 93/12085 Al)
• Kende, et al described methodology for preparing 3,3-substituted oxindoles, e g X, that was utilized in the present invention (Synth Commun , 12, 1, 1982)
• U.S Patent No. 5,521,166 teaches cyclophasic hormonal regimens comprising an antiprogestin and a progestin wherein the progestin is administered in the alternating presence and absence of an antiprogestin.
• the disclosed regimens also provide for use of an estrogen for a period of from 2-4 days to prevent breakthrough bleeding.
• This invention provides combination therapies and dosing regimens utilizing antiprogestational agents in combination with one or more progestational agents
• This invention further provides methods of treatment and dosmg regimens further utilizing in combination with these antiprogestins and progestins, an estrogen, such as ethinyl estradiol
• These regimens and combinations may be administered to a mammal to induce contraception or for the treatment and/or prevention of secondary amenorrhea, dysfunctional bleeding, uterine leiomyomata, endometriosis, polycystic ovary syndrome, carcinomas and adenocarcinomas of the endomet ⁇ um, ovary, breast, colon, prostate Additional uses of the invention mclude stimulation of food intake
• the uses herein for the treatment and/or prevention of the conditions or diseases described above includes the continuous administration or periodic discontinuation of administration of the invention to allow for minimization of effect dose or minimization of side effects or cyclic menstrual bleeding
• the use of this invention for contraception includes administration, preferably orally, to a female of child bearing age an antiprogestin in combination with an estrogen or progestin or both
• These administration regimens are preferably carried out over 28 consecutive days, with a terminal portion of the cycle containing administration of no progestins, estrogens or anti-progestins
• the progestins of this invention may be administered alone or in combination with estrogen for the initial 18 to 21 days of a 28-day cycle, followed by administration of an antiprogestin, alone or in combination with an estrogen, for from 1 to 7 days.
• the estrogen to be used in the combinations and formulations of this invention is preferably ethinyl estradiol.
• Progestational agents useful with this invention include, but are not limited to, levonorgestrel, norgestrel, desogestrel, 3-ketodesogestrel, norethindrone, gestodene, norethindrone acetate, norgestimate, osaterone, cyproterone acetate, trimegestone, dienogest, drospirenone, nomegestrol, or (17-deacetyl)norgestimate .
• progestins for use in the combinations of this invention are levonorgestrel, gestodene and trimegestone.
• Examples of orally administered regimens of this invention over a 28 day cycle include administration of a progestational agent solely for the first 21 days at a daily dose equal in progestational activity to from about 35 to about 100 ⁇ g of levonorgestrel.
• An antiprogestin compound of this invention may then be administered at a daily dose of from about 2 to 50 mg from day 22 to day 24, followed by no administration or administration of a placebo for days 25 to 28. It is most preferred that the daily dosages of each relevant active ingredient be incorporated into a combined, single daily dosage unit, totaling 28 daily units per 28- day cycle.
• a progestational agent may be coadministered for the first 21 days at a daily dose equal in progestational activity to from about 35 to about 150 ⁇ g levonorgestrel, preferably equal in activity to from about 35 to about 100 ⁇ g levonorgestrel, with an estrogen, such as ethinyl estradiol, at a daily dose range of from about 10 to about 35 ⁇ g.
• an antiprogestin administered at a daily dose of from about 2 to 50 mg from day 22 to day 24, followed by no administration or administration of a placebo for days 25 to 28.
• Still another regimen within the scope of this invention will include coadministration from days 1 to 21 of a progestational agent, the progestational agent, preferably levonorgestrel, being administered at a daily dose equal in progestational activity to from about 35 to about 100 ⁇ g levonorgestrel, and an estrogen, such as ethinyl estradiol, at a daily dose range of from about 10 to about 35 ⁇ g.
• a progestational agent preferably levonorgestrel
• an estrogen such as ethinyl estradiol
• kits or packages of pharmaceutical formulations designed for use in the regimens described herein are preferably designed for daily oral administration over a 28-day cycle, preferably for one oral administration per day, and organized so as to indicate a single oral formulation or combination of oral formulations to be taken on each day of the 28-day cycle.
• each kit will include oral tablets to be taken on each the days specified, preferably one oral tablet will contain each of the combined daily dosages indicated.
• one 28-day kit may comprise: a) an initial phase of from 14 to 21 daily dosage units of a progestational agent equal in progestational activity to about 35 to about 150 ⁇ g levonorgestrel, preferably equal in progestational activity to about 35 to about 100 ⁇ g levonorgestrel; b) a second phase of from 1 to 11 daily dosage units of an antiprogestin compound of this invention, each daily dosage unit containing an antiprogestin compound at a daily dosage of from about 2 to 50 mg; and c) 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.
• a preferred embodiment of this kit may comprise: a) an initial phase of 21 daily dosage units of a progestational agent equal in progestational activity to about 35 to about 150 ⁇ g levonorgestrel, preferably equal in progestational activity to about 35 to about 100 ⁇ g levonorgestrel; b) a second phase of 3 daily dosage units for days 22 to 24 of an antiprogestin compound of this invention, each daily dosage unit containing an antiprogestin compound at a daily dosage of from about 2 to 50 mg; and c) optionally, a third phase of 4 daily units of an orally and pharmaceutically acceptable placebo for each of days 25 to 28.
• Another 28-day cycle packaging regimen or kit of this invention comprises: a) a first phase of from 18 to 21 daily dosage units of a progestational agent equal in progestational activity to about 35 to about 150 ⁇ g levonorgestrel, preferably equal in activity to from about 35 to about 100 ⁇ g levonorgestrel, and, as an estrogen, ethinyl estradiol at a daily dose range of from about 10 to about 35 ⁇ g; and b) a second phase of from 1 to 7 daily dosage units of an antiprogestin of this invention at a daily dose of from about 2 to 50 mg; and c) optionally, an orally and pharmaceutically acceptable placebo for each of the remaining 0-9 days in the 28-day cycle in which no progestational agent, estrogen or antiprogestin is administered.
• a preferred embodiment of the kit described above may comprise: a) a first phase of 21 daily dosage units of a progestational agent equal in progestational activity to about 35 to about 150 ⁇ g levonorgestrel, preferably equal in activity to from about 35 to about 100 ⁇ g levonorgestrel, and, as an estrogen, ethinyl estradiol at a daily dose range of from about 10 to about 35 ⁇ g; and b) a second phase of 3 daily dosage units for days 22 to 24 of an antiprogestin administered at a daily dose of from about 2 to 50 mg; and c) optionally, a third phase of 4 daily dose units of an orally and pharmaceutically acceptable placebo for each of days 25 to 28.
• a further 28-day packaged regimen or kit of this invention comprises. a) a first phase of from 18 to 21 daily dosage units, each containing a progestational agent of this invention at a daily dose equal in progestational activity to about 35 to about 150 ⁇ g levonorgestrel, preferably equal in activity to from about 35 to about 100 ⁇ g levonorgestrel, and ethinyl estradiol at a daily dose range of from about 10 to about 35 ⁇ g; b) a second phase of from 1 to 7 daily dose units, each daily dose unit containing an antiprogestin of this invention at a concentration of from 2 to 50 mg, and ethinyl estradiol at a concentration of from about 10 to about 35 ⁇ g; and c) optionally, an orally and pharmaceutically acceptable placebo for each of the remaining 0-9 days in the 28-day cycle in which no progestational agent, estrogen or antiprogestin is administered.
• a preferred embodiment of the package or kit just described comprises. a) a first phase of 21 daily dosage units, each containing a progestational agent of this invention at a daily dose equal in progestational activity to about 35 to about 150 ⁇ g levonorgestrel, preferably from about 35 to about 100 ⁇ g levonorgestrel, and ethinyl estradiol at a daily dose range of from about 10 to about 35 ⁇ g, b) a second phase of 3 daily dose units for days 22 to 24, each dose unit containing an antiprogestin of this invention at a concentration of from 2 to 50 mg, and ethinyl estradiol at a concentration of from about 10 to about 35 ⁇ g, and c) optionally, a third phase of 4 daily units of an orally and pharmaceutically acceptable placebo for each of days 25 to 28.
• each pharmaceutically active component of the regimen remains fixed m each particular phase in which it is administered
• the daily dose units described are to be admmistered in the order described, with the first phase followed in order by the second and third phases.
• the kits contain the placebo described for the final days of the cycle
• each package or kit comp ⁇ se a pharmaceutically acceptable package having indicators for each day of the 28-day cycle, such as a labeled blister package or dial dispenser packages known in the art
• anti-progestational agents anti-progestational agents
• anti-progestins anti-progestins and progesterone receptor antagonists
• progestins progestational agents
• progesterone receptor agonists are understood to refer to compounds of the same activity
• dosage regimens may be adjusted to provide the optimal therapeutic response
• several divided doses of each component may be administered daily or the dose may be proportionally increased or reduced as indicated by the exigencies of the therapeutic situation
• reference to a daily dosage unit may also include divided units which are administered over the course of each day of the cycle contemplated
• R, and R 2 are chosen independently from H, alkyl, substituted alkyl, OH,
• R 2 are joined to form a ring comprising one of the following
• R, and R 2 comprise a double bond to CMe 2 , C(cycloalkyl), O, or C(cyloether); n is an integer from 0 to 5; m is an integer from 1 to 4; p is an integer from 1 to 4;
• R 3 is selected from H, OH, NH 2 , C, to C 6 alkyl, substituted C, to C 6 alkyl, C 3 to C 6 alkenyl, alkynyl or substituted alkynyl, or COR A ;
• R A is selected from H, C, to C 3 alkyl, substituted C, to C 3 alkyl, C, to C 3 alkoxy, substituted C, to C 3 alkoxy, C, to C 3 aminoalkyl, or substituted C, to C 3 aminoalkyl;
• R 4 is selected from H, halogen, CN, NH 2 , C, to C 6 alkyl, substituted C, to C 6 alkyl, C, to C 6 alkoxy, substituted C, to C 6 alkoxy, C, to C 6 aminoalkyl, or substituted C, to C 6 aminoalkyl;
• R 5 is selected from the groups a), b) or c): a) R 5 is a trisubstituted benzene ring containing the substituents X, Y and Z as shown below:
• X is selected from the group of halogen, OH, CN, C, to C 3 alkyl, substituted C, to C 3 alkyl, C, to C 3 alkoxy, substituted C, to C 3 alkoxy, C, to C 3 thioalkyl, substituted C, to C 3 thioalkyl, S(0)alkyl, S(O) 2 alkyl, C, to C 3 aminoalkyl, substituted C, to C 3 aminoalkyl, NO 2 , C, to C 3 perfluoroalkyl, 5 or 6 membered heterocyclic ring containing 1 to 3 heteroatoms, COR B , OCOR B , or NR c COR B ;
• R B is H, C, to C 3 alkyl, substituted C, to C 3 alkyl, aryl, substituted aryl, C, to C 3 alkoxy, substituted C, to C 3 alkoxy, C, to C 3 aminoalkyl, or substituted C, to C 3
• R c is H, C, to C 3 alkyl, or substituted C, to C 3 alkyl
• Y and Z are independently selected from H, halogen, CN, N0 2 , C, to C 3 alkoxy, C, to C 3 alkyl, or C, to C 3 thioalkyl
• R 5 is a five or six membered heterocyclic ring with 1 , 2, or 3 heteroatoms selected from O, S, SO, S0 2 or NR 6 and containing one or two independent substituents from the group of H, halogen, CN, N0 2 and C, to C 3 alkyl, C, to C 3 alkoxy, C, to C 3 aminoalkyl, COR D , or NR E COR D ,
• R D is H, C, to C 3 alkyl, substituted C, to C 3 alkyl, aryl, substituted aryl, C, to C 3 alkoxy, substituted C, to C 3 alkoxy, C, to C 3 aminoalkyl, or substituted C, to C 3 aminoalkyl
• R k is H, C, to C 3 alkyl, or substituted C, to C 3 alkyl
• R 6 is H, or C, to C 3 alkyl, or c) R 5 is an mdol-4-yl, ⁇ ndol-7-yl or benzo-2-th ⁇ ophene moiety, the moiety being optionally substituted by from 1 to 3 substituents selected from halogen, lower alkyl, CN, N0 2 , lower alkoxy, or CF 3 , or a pharmaceutically acceptable salt thereof
• R 5 is a disubstituted benzene ring containing the substituents X and Y as shown below:
• X is taken from the group of halogen, CN, C, to C 3 alkoxy, C, to C 3 alkyl, NO,, C, to C 3 perfluoroalkyl, 5 membered heterocyclic ring containing 1 to 3 heteroatoms or C, to C 3 thioalkoxy,
• Y is a substituent on the 4' or 5 'position of the disubstituted benzene ring selected from the group of H, halogen, CN, N0 2 , C, to C 3 alkoxy, C, to C 4 alkyl, or C, to C 3 thioalkyl; or a pharmaceutically acceptable salt thereof
• Another preferred group of this invention comprises compounds of formulas 2 and 2a wherein R 5 is a five membered ring with the structure shown below
• U is 0, S, or NR 6 ,
• R 6 is H, or C, to C 3 alkyl, C, to C 4 CO-alkyl,
• X' is selected from the group of halogen, CN, NO,, C, to C 3 alkyl or C, to C 3 alkoxy, with a proviso that, when X' is CN, U is not NR 6 , Y' is selected from H, F, CN, N0 2 or C, to C 4 alkyl, or a pharmaceutically acceptable salt thereof
• R 3 is a six membered ring with the structure shown
• X 1 is N or CX 2 ,
• X 2 is halogen, CN or NO 2 ; or pharmaceutically acceptable salt thereof
• the compounds of this invention may contain an asymmetric carbon atom and some of the compounds of this invention may contain one or more asymmetric centers and may thus give rise to optical isomers and diastereomers. While shown without respect to stereochemistry in Formula 1 and 2 the present invention includes such optical isomers and diastereomers; as well as the racemic and resolved, enantiomerically pure R and S stereoisomers; as well as other mixtures of the R and S stereoisomers and pharmaceutically acceptable salts thereof.
• alkyl is used herein to refer to both straight- and branched-chain saturated aliphatic hydrocarbon groups having 1 to 8 carbon atoms; "alkenyl” is intended to include both straight- and branched-chain alkyl group with 1 or 2 carbon- carbon double bonds and containing 2 to 8 carbon atoms; “alkynyl” group is intended to cover both straight- and branched-chain alkyl group with at least 1 or 2 carbon-carbon triple bonds and containing 2 to 8 carbon atoms.
• substituted alkyl refers to alkyl, alkenyl, and alkynyl as just described having one or more substituents from the group including halogen, CN, OH, NO 2 , amino, aryl, heterocyclic, substituted aryl, substituted heterocyclic, alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, arylthio.
• substituents may be attached to any carbon of alkyl, alkenyl, or alkynyl group provided that the attachment constitutes a stable chemical moiety.
• aryl is used herein to refers to an aromatic system which may be a single ring or multiple aromatic rings fused or linked together as such that at least one part of the fused or linked rings forms the conjugated aromatic system.
• the aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, anthryl, tetrohydronaphthyl, phenanthryl.
• substituted aryl refers to aryl as just defined having 1 to 4 substituents from the group including halogen, CN, OH, NO 2 , amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, or arylthio.
• heterocyclic is used herein to describe a stable 4- to 7-membered monocyclic or a stable multicyclic heterocyclic ring which is saturated, partially unsaturated, or unsaturated, and which consists of carbon atoms and from one to four heteroatoms selected from the group including N, O, and S atoms.
• the N and S atoms may be oxidized.
• the heterocyclic ring also includes any multicyclic ring in which any of above defined heterocyclic rings is fused to an aryl ring.
• the heterocyclic ring may be attached at any heteroatom or carbon atom provided the resultant structure is chemically stable.
• heterocyclic groups include, but are not limited to, tetrahydrofuran, piperidinyl, piperazinyl, 2-oxopiperidinyl, azepinyl, pyrrolidinyl, imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, isoxazolyl, morpholinyl, indolyl, quinolinyl, thienyl, furyl, benzofuranyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, and isoquinolinyl.
• substituted heterocyclic is used herein to describe the heterocyclic just defined having 1 to 4 substituents selected from the group which includes halogen, CN, OH, N0 2 , amino, alkyl, substituted alkyl, cycloalkyl, alkenyl, substituted alkenyl, alkynyl, alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, or arylthio.
• thioalkyl is used herein to refer to the SR group, where R is alkyl or substituted alkyl, containing 1 to 8 carbon atoms, preferably 1 to 6 carbon atoms.
• alkoxy is used herein to refer to the OR group, where R is alkyl or substituted alkyl, containing 1 to 8 carbon atoms, preferably 1 to 6 carbon atoms.
• aryloxy is used herein to refer to the OR group, where R is aryl or substituted aryl, as defined above.
• alkylcarbonyl is used herein to refer to the RCO group, where R is alkyl or substituted alkyl, containing 1 to 8 carbon atoms, preferably 1 to 6 carbon atoms.
• alkylcarboxy is used herein to refer to the COOR group, where R is alkyl or substituted alkyl, containing 1 to 8 carbon atoms, preferably 1 to 6 carbon atoms.
• aminoalkyl refers to both secondary and tertiary amines wherein the alkyl or substituted alkyl groups, containing 1 to 8 carbon atoms, preferably 1 to 6 carbon atoms, which may be either the same or different and the point of attachment is on the nitrogen atom.
• halogen refers to Cl, Br, F, or I.
• anti-progestational compounds of this invention may be prepared according to the methods described below.
• oxindole 5 is treated with mixture a strong organo-metallic base (e.g. butyl lithium, lithium diisopropylamide, potassium hexamethyldisilazide) in an inert solvent (e.g. THF, diethyl ether) under nitrogen at reduce temperature (ca. -20 °C) (Kende, et al, Synth. Commun., 12, 1, 1982).
• a strong organo-metallic base e.g. butyl lithium, lithium diisopropylamide, potassium hexamethyldisilazide
• an inert solvent e.g. THF, diethyl ether
• excess electrophile such as an alkyl halide, preferably the iodide. If R, and R 2 are to be joined such as the product 6 contains a spirocycle at position 3, then the electrophile should be bifunctional, i.e.
• bromide 7 is reacted with a palladium salt (e.g. tetrakis(triphenylphoshine)palladium(0)), in a suitable solvent (e.g. THF, dimethoxyethane, ethanol, toluene) at room temperature under an inert atmosphere (argon, nitrogen).
• a palladium salt e.g. tetrakis(triphenylphoshine)palladium(0)
• a suitable solvent e.g. THF, dimethoxyethane, ethanol, toluene
• the mixture is then treated with an arylboronic acid or boronic acid ester and a base (sodium carbonate, triethylamine, potassium phosphate) in water or fluoride source (cesium fluoride) under anhydrous conditions.
• a base sodium carbonate, triethylamine, potassium phosphate
• fluoride source cesium fluoride
• the resultant mono-alkylated compound may be then isolated and re-subjected to the reaction conditions using R 2 -X, or alternatively used in-situ for the second alkylation with R 2 -X.
• an organo zinc or magnesium reagent from compound 7 and react it in-situ with an aryl bromide, aryl iodide, aryltrifluoromethane sulfonate of arylfluorosulfonate, under palladium catalyzed conditions to afford compound 8
• an organo zinc or magnesium species could be prepared by treatment of the bromide 7 in an anhydrous solvent (e g THF, Et 2 0) with a strong base (sodium hydride preferred, sodium hexamethyldisilazide, potassium hydride) followed by reaction at reduced temperature (-50 to -20 °C) with n- butyllithium and N,N,N',N'-tetramethylethylened ⁇ am ⁇ ne followed after a suitable period of time by reaction with anhydrous zinc chloride or magnesium bromide
• an anhydrous solvent e g THF, Et 2 0
• a strong base sodium hydride preferred, sodium hexamethyld
• the anti-progestational compounds of the present invention can be used m the form of salts derived from pharmaceutically or physiologically acceptable acids or bases
• These salts include, but are not limited to, the following salts with inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and, as the case may be, such organic acids as acetic acid, oxalic acid, succimc acid, and maleic acid
• Other salts include salts with alkali metals or alkaline earth metals, such as sodium, potassium, calcium or magnesium in the form of esters, carbamates and other conventional "pro-drug" forms, which, when administered in such form, convert to the active moiety in vivo
• the compounds and combinations herein may be combined with one or more pharmaceutically acceptable carriers or excipients, for example, solvents, diluents and the like, and may be administered orally in such forms as tablets, capsules, dispersible powders, granules, or suspensions containing, for example, from about 0 05 to 5% of suspending agent, syrups containing, for example, from about 10 to 50% of sugar, and elixirs containing, for example, from about 20 to 50% ethanol, and the like, or parenterally in the form of sterile injectable solutions or suspensions containing from about 0 05 to 5% suspending agent in an isotomc medium
• Such pharmaceutical preparations may contain, for example, from about 25 to about 90%) of the active ingredient in combination with the carrier, more usually between about 5% and 60% by weight
• Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include sterile water, polyethylene glycols, non-ionic surfactants and edible oils such as corn, peanut and sesame oils, as are appropriate to the nature of the active ingredient and the particular form of administration desired
• Adjuvents customarily employed in the preparation of pharmaceutical compositions may be advantageously included, such as flavoring agents, coloring agents, preserving agents, and antioxidants, for example, vitamin E, ascorbic acid, BHT and BHA
• compositions from the standpoint of ease of preparation and administration are solid compositions, particularly tablets and hard-filled or liquid-filled capsules Oral administration of the compounds is preferred
• solutions or suspensions of these active compounds as a free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose Dispersions can also be prepared in glycerol, liquid, polyethylene glycols and mixtures thereof in oils Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms
• the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions
• the form must be sterile and must be fluid to the extent that easy syringe ability exits It must be stable under conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacterial and fungi
• the carrier can be a solvent or dispersion medium containing, for example, water, ethanol (e g , glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oil
• the present invention may be further understood by the following non- hmiting examples of anti-progestational compounds
• reaction mixture was treated with 20 mL of sodium bicarbonate solution and was then extracted with ethyl acetate (2x5 OmL). The combined organic phases were washed with saturated brine, dried (MgS0 4 ). Recrystallization from ethanol gave 0.13 g of pure product, mp 227 - 228 °C.
• 5-Bromo-2-thiophenecarbonitrile A mixture of 5-bromo-2- thiophenecarboxaldehyde (96. Og, 500 mmol), hydroxylamine hydrochloride (1 1 1.9 g, 500 mmol), pyridine (500 mL), and ethanol (500 mL) was heated under nitrogen at reflux for two hours. The reaction mixture was cooled to ambient temperature and concentrated in vacuo to give an oil. The crude product was triturated twice with ice water and the solid obtained was collected on a filter.
• EXAMPLE 60 4-(l,2-Dihvdro-2-oxospiro[cvclohexane-l,3-[3Hlindoll-5-yl)-2- furancarbonitrile A solution of 3-bromo-5-cyano-furan (0J5 g, 4.4 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.4 g) in ethylene glycol dimethyl ether (20 cm 3 ) was stirred under N 2 for 20 minutes.
• the title compound was prepared from (2'-oxo-2,3-dihydrospiro[cyclohexane- l,3'-[3H]indol]-5'-yl)boronic acid (2.5 g, 10 mmol) and 5-bromo-2-fluoro- trifluoromethylbenzene (2 g, 8 mmol) as described for example 18, to afford the title compound (0.87 g, 30%) as a solid: mp.
• EXAMPLE 66 5-(3-Chloro-4-fluoro-phenv ⁇ -3,3-dimethyl-l,3-dihydro-indol-2-one A solution of 5-bromo-3,3-dimethyl-l,3-dihydro-indol-2-one (0.35 g, 1.46 mmol) and tetrakis(triphenylphosphine) palladium (0.13 g, 0.11 mmol) in dimethoxyethane (10 cm3) was stirred under N 2 for 20 min..
• EXAMPLE 70 5-(3-Chloro-5-fluoro-phenyl)-3,3-dimethyl-l,3-dihydro-indol-2-one Prepared according to the procedure for example 18 for using (2'-oxo-[2, 3- dihydro-3,3-dimethyl -1, 3'- [3H] indol] -5'-yl) boronic acid (345 mg, 1.7 mmol) and l-bromo-3-chloro-5-fluorobenzene (295 mg, 1.4 mmol) to afford the title compound (245 mg, 0.85 mmol, 60 %) as a white solid: mp. 205.9-206.8 °C .
• EXAMPLE 74 S'- ⁇ -Cyano ⁇ -fluorophenyD-spirofcyclopentane-l ⁇ '-PHlindoll ⁇ 'd'H one A solution of 4-cyano-3-fluoro-bromobenzene (0.63 g, 3.1 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.2 g) in ethylene glycol dimethyl ether (20 cm 3 ) was stirred under N 2 for 20 minutes.
• In-vitro potencies lie in the range 0.01 nM - 10,000 nM, and in-vivo potencies in the range 1 ⁇ g/kg to 100 mg/kg.
• the in-vitro biology is determined by (1) competitive Radioligand Binding: using the A- form of the human progesterone receptor with progesterone as the radioligand; (2) co-transfection assay, which provides functional activity expressed as agonist EC50 and Antagonist IC50 values; (3) a T47D cell proliferation, which is a further functional assay which also provides agonist and antagonist data; and (4) T47D cell alkaline phosphatase assay , which is a further functional assay which also provides agonist and antagonist data.
• hPR Binding assay This assay is carried out in accordance with: Pathirana, C; Stein, R.B.; Berger, T.S.; Fenical, W.; Ianiro, T.; Mais, D.E.; Torres, A.; Glodman, M.E., Nonsteroidal human progesterone receptor modulators from the marine alga cymoplia barbata, J. Steroid Biochem. Mol. Biol., 1992, 41, 733-738.
• the object of this assay is to determine a compound's progestational or antiprogestational potency based on its effect on PRE-luciferase reporter activity in CV-1 cells co-transfected with human PR and PRE-luciferase plasmids.
• the materials methods used in the assay are as follows.
• a. Medium The growth medium was as follows: DMEM (BioWhittaker) containing 10% (v/v) fetal bovine serum (heat inactivated), 0.1 mM MEM non-essential amino acids, lOOU/ml penicillin, lOOmg/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL).
• the experimental medium was as follows: DMEM (BioWhittaker), phenol red-free, containing 10%) (v/v) charcoal- stripped fetal bovine serum (heat-inactivated), 0.1 mM MEM non-essential amino acids, lOOU/ml penicillin, lOOmg/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL).
• DMEM BioWhittaker
• phenol red-free containing 10%
• fetal bovine serum fetal bovine serum
• 0.1 mM MEM non-essential amino acids lOOU/ml penicillin
• lOOmg/ml streptomycin lOOmg/ml streptomycin
• 2 mM GlutaMax mM GlutaMax
• Stock CV-1 cells are maintained in growth medium. Co-transfection is done using 1.2xl0 7 cells, 5 mg pLEM plasmid with hPR-B inserted at Sphl and BamHl sites, 10 mg pGL3 plasmid with two PREs upstream of the luciferase sequence, and 50 mg sonicated calf thymus DNA as carrier DNA in 250 ml. Electroporation is carried out at 260 V and 1,000 mF in a Biorad Gene Pulser II. After electroporation, cells are resuspended in growth medium and plated in 96-well plate at 40,000 cells/well in 200 ⁇ l. Following overnight incubation, the medium is changed to experimental medium.
• Cells are then treated with reference or test compounds in experimental medium. Compounds are tested for antiprogestational activity in the presence of 3 nM progesterone. Twenty-four hr. after treatment, the medium is discarded, cells are washed three times with D-PBS (GIBCO, BRL). Fifty ⁇ l of cell lysis buffer (Promega, Madison, WI) is added to each well and the plates are shaken for 15 min in a Titer Plate Shaker (Lab Line Instrument, Inc.). Luciferase activity is measured using luciferase reagents from Promega. c. Analysis of Results:
• Each treatment consists of at least 4 replicates.
• Log transformed data are used for analysis of variance and nonlinear dose response curve fitting for both agonist and antagonist modes. Huber weighting is used to downweight the effects of outliers.
• EC 50 or IC 50 values are calculated from the retransformed values.
• JMP software SAS Institute, Inc. is used for both one-way analysis of variance and non-linear response analyses.
• Progesterone and trimegestone are reference progestins and RU486 is the reference antiprogestin. All reference compounds are run in full dose- response curves and the EC 50 or IC 50 values are calculated.
• 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)
• EC 50 Concentration of a compound that gives half-maximal increase PRE- luciferase activity (default-nM) with SE.
• IC 50 Concentration of a compound that gives half-maximal decrease in 3 nM progesterone induced PRE-luciferase activity (default-nM) with SE.
• T47D cell proliferation assay The objective of this assay is the determination of progestational and antiprogestational potency by using a cell proliferation assay in T47D cells. A compound's effect on DNA synthesis in T47D cells is measured.
• the materials and methods used in this assay are as follows. a. Growth medium: DMEM:F12 (1 : 1) (GIBCO, BRL) supplemented with 10% (v/v) fetal bovine serum (not heat- inactivated), lOOU/ml penicillin, lOOmg/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL). b.
• Treatment medium Minimum Essential Medium (MEM) (#51200-038GIBCO, BRL) phenol red-free supplemented with 0.5% charcoal stripped fetal bovine serum, lOOU/ml penicillin, 200 mg/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL).
• MEM Minimum Essential Medium
• lOOU/ml penicillin 200 mg/ml streptomycin
• 2 mM GlutaMax mM GlutaMax
• BrdU inco ⁇ oration is measured using a cell proliferation ELISA kit (#RPN 250, Amersham Life Science) according to manufacturer's instructions. Briefly, cells are fixed in an ethanol containing fixative for 30 min, followed by incubation in a blocking buffer for 30 min to reduce background. Peroxidase-labeled anti-BrdU antibody is added to the wells and incubated for 60 min. The cells are rinsed three times with PBS and incubated with 3,3'5,5'-tetramethylbenzidine (TMB) substrate for 10-20 min depending upon the potency of tested compounds. Then 25 ⁇ l of 1 M sulfuric acid is added to each well to stop color reaction and optical density is read in a plate reader at 450 nm within 5 min. e. Analysis of Results:
• the pu ⁇ ose of this assay is to identify progestins or antiprogestins by determining a compound's effect on alkaline phosphatase activity in T47D cells.
• the materials and methods used in this assay are as follows. a. Culture medium: DMEM:F12 (1 : 1 ) (GIBCO. BRL) supplemented with 5%> (v/v) charcoal stripped fetal bovine serum (not heat-inactivated), lOOU/ml penicillin, 100 ⁇ g/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL).
• 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 ⁇ l of diluted cell suspension was added. Twenty ⁇ l of reference or test compounds diluted in the culture medium was then added to each well. When testing for progestin antagonist activity, reference antiprogestins or test compounds were added in the presence of 1 nM progesterone. The cells were incubated at 37°C in a 5% C0 2 /humidified atmosphere for 24 hr. d. Alkaline Phosphatase Enzyme Assay:
• Huber weighting is used to downweight the effects of outliers.
• EC 50 or IC 50 values are calculated from the retransformed values.
• JMP software SAS Institute, Inc.
• JMP software is used for both one-way analysis of variance and non-linear dose response analyses in both single dose and dose response studies.
• Progesterone and trimegestone are reference progestins and RU486 is the reference antiprogestin. All reference compounds are run in full dose response curves and the EC 50 or IC 50 values are calculated.
• the primary in-vivo assay is the rat deciduahzation model which may be used to determine progestational effects of both agonists and antagonists
• the secondary m-vivo assay is the rat ovulation inhibition model which is under development and hence the protocol is unavailable.
• Rat deciduahzation assay The objective of this procedure is used to evaluate the effect of progestins and antiprogestins on rat uterine deciduahzation and compare the relative potencies of various test compounds.
• the materials and methods used in this assay are as follows. a. Methods: Test compounds are dissolved in 100% ethanol and mixed with com oil (vehicle). Stock solutions of the test compounds in oil (MazolaTM) are then prepared by heating (-80 °C) the mixture to evaporate ethanol. Test compounds are subsequently diluted with 100% com oil or 10%> ethanol in com 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 230g) are obtained from Taconic (Taconic Farms, NY) following surgery. Ovariectomy is performed at least 10 days prior to treatment to reduce circulating sex steroids. Animals are housed under 12 hr light/dark cycle and given standard rat chow and water ad libitum. c. Treatment Rats are weighed and randomly assigned to groups of 4 or 5 before treatment. Test compounds in 0.2 ml vehicle are administered by subcutaneous injection in the nape of the neck or by gavage using 0.5 ml. The animals are treated once daily for seven days.
• mice are given the test compounds and a EC50 dose of progesterone (5.6 mg/kg) during the first three days of treatment. Following decidual stimulation, animals continue to receive progesterone until necropsy four days later, d. Dosing
• Doses are prepared based upon mg/kg mean group body weight. In all studies, a control group receiving vehicle is included. Determination of dose-response curves is carried out using doses with half log increases (e.g. 0.1, 0.3, 1.0, 3.0 mg/kg). e. Decidual induction
• deciduahzation is induced in one of the uterine horns by scratching the antimesometrial luminal epithelium with a blunt 21 G needle.
• the contralateral horn is not scratched and serves as an unstimulated control.
• rats are sacrificed by C ⁇ 2 asphyxiation and body weight measured. Uteri are removed and trimmed of fat.
• Decidualized (D-hom) and control (C-hom) uterine horns are weighed separately.
• the increase in weight of the decidualized uterine hom is calculated by D-horn/C-horn and logarithmic transformation is used to maximize normality and homogeneity of variance.
• the Huber M-estimator is used to down weight the outlying transformed observations for both dose-response curve fitting and one-way analysis of variance.
• JMP software SAS Institute, Inc.
• AS Institute, Inc. is used for both oneway ANOVA and non-linear dose-response analyses.
• D-hom Wet weight of decidualized uterine hom (default-mg)
• Progestational activity Compounds that induce deciduahzation significantly (p ⁇ 0.05) compared to vehicle control are considered active
• Antiprogestational activity Compounds that decrease EC 50 progesterone induced deciduahzation significantly (p ⁇ 0.05)
• EC 50 for uterine weight Concentration of compound that gives half-maximal increase in decidual response (default-mg/kg)
• IC 50 for uterine weight Concentration of compound that gives half-maximal decrease in EC 50 progesterone induced decidual response (default-mg/kg)
• reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, washed with brine (50 mL) and dried over magnesium sulfate. The solution was filtered, concentrated in vacuo, and the residue was purified by flash column chromatography to give the product (0J g, 86%) as a tan powder, mp 163-165 °C.
• reaction mixture was diluted with dichloromethane (100 mL) and filtered through celite. The filtrate was poured into water (100 mL) and the layers were separated. The organic layer was washed with brine (50 mL) and dried over magnesium sulfate. The solution was filtered, concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (40%) ethyl acetate/hexane) to give the title compound (415 mg, 37%) as a yellow powder, mp 265 °C (dec).
• reaction mixture was poured into water (50 mL) and extracted with ethyl acetate (2 x 50 mL). The organic layers were combined, washed with brine (50 mL), dried over magnesium sulfate, filtered and concentrated in vacuo.
• the title compound was prepared in a manner similar to example 69 from 5- bromo-4-n-butyl thiophenecarbonitrile 1 (1.24 g, 5.1 mmol), (1,2-dihydro -2- oxospiro[cyclohexane-l,3-[3H] indol)-5-boronic acid (1.24 g, 5.05 mmol), tetrakis(triphenylphosphine) palladium (0.25 g), potassium carbonate (2.75 g, 21 mmol), water (10 mL),and dimethoxyethane (50 mL) heated at reflux for 5 hours to afford the title compound (1 g, 54%), m.p.130- 132° C.
• 5-(3-chlorophenyl)-4-methylspiro[cvclohexane-l,3-r3H]indol1-2(lH)-one A solution of the above 5-bromo-4-methyl-oxindole (0.1 g, 0.34 mmol) and tetrakis(triphenylphosphine) palladium (0.05 g, 0.04 mmol) in dimethoxyethane (10 mL) was stirred under N 2 for 20 min. To this mixture was then added 3- chlorophenylboronic acid (0.065 g, 0.41 mmol) and sodium carbonate (0.1 g, 1.0 mmol) in water (3 mL).
• the solution was removed from the cooling bath and allowed to reach room temperature.
• the solution was filtered through celite, concentrated in vacuo and redissolved in MeOH (10 mL). A large excess of ammonium acetate was added and the solution was heated to 60 °C for lh then stored in a refrigerator for 16h.
• the solution was partitioned between DCM and water.
• the title compound was prepared from CAT-817819 (1.9 g, 7.8 mmol ) and 2- bromochlorobenzene (1.0 g, 5.2 mmol) according to the method for Example 18 to afford the title compound (0.68 g, 42%) as an off white solid: mp.

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