WO2004037792A2

WO2004037792A2 - Benzo (f) quinolinone variants as nuclear hormone receptor ligands

Info

Publication number: WO2004037792A2
Application number: PCT/EP2003/010108
Authority: WO
Inventors: Marcus Ruda; David Wensbo
Original assignee: Karo Bio Ab
Priority date: 2002-09-27
Filing date: 2003-09-10
Publication date: 2004-05-06
Also published as: AU2003293305A1; WO2004037792A3; GB0222516D0; AU2003293305A8

Abstract

The present invention relates to compounds and derivatives thereof, their synthesis, and their use as nuclear hormone receptor modulators. The compounds of the instant invention are ligands for nuclear hormone receptors and as such may be useful for treatment or prevention of a variety of conditions related to nuclear hormone functioning including proliferative diseases, cancers, benign prostate hyperplasia, adenomas and neoplasisms of the prostate, benign or malignant tumor cells containing the androgen receptor, cardiovascular diseases, angiogenic conditions or disorders, hirsutism, acne, hyperpilosity, inflammation, immune modulation, seborrhea, endometriosis, polycycstic ovary syndrome, androgen alopecia, hypogonadism, osteoporosis, suppressing spermatogenesis, libido, cachexia, anorexia, cancers expressing the estrogen receptor, prostate cancer, breast cancer, endometrial cancer, incontinence, hot flashes, vaginal dryness, menopause, contraception, pregnancy termination, cancers containing the progesterone e receptor, endometriosis, cachexia, menopause fibrosis, labor induction, autoimmune diseases, impairment of cognitive functioning, cerebral degenerative disorders, Alzheimer's disease, psychotic disorders, type-II diabetes, congestive heart failure, disregulation of cholesterol or lipid homeostasis, and/or obesity. A compound having the general formula I or II:

Description

Novel Nuclear Hormone Receptor Ligands and Methods,

HELD OF INVENTION This invention relates to novel fused pyridone compounds which are nuclear hormone receptor ligands and are preferably selective for either the androgen, estrogen, glucocorticoid, liver X receptor, mineralocorticoid, peroxisome proliferator-activated, or progesterone receptor, to methods of preparing such compounds and to methods for using such compounds for treating nuclear hormone associated diseases.

BACKGROUND OF INVENTION Nuclear hormone receptors (NHR's) constitute a large super-family of ligand-dependent and DNA sequence specific transcription factors. Members of this family influence transcription either directly, through specific binding to the promoter target genes (Evans, Science 240: 889-895 (1988)), or indirectly, via protein-protein interactions with other transcription factors (Jonat et al., Cell 62:1189-1204 (1990), and Yang- Yen et al., Cell 62:1205-1215 (1990)). The nuclear hormone receptor super-family (also known in the art as the "steroid/thyroid hormone receptor super-family") includes receptors for a variety of hydrophobic ligands including cortisol, aldosterone, estrogen, progesterone, testosterone, vitamin D₃, thyroid hormone T₃, and retinoic acid (Evans, 1988, vide supra). In addition to these conventional nuclear hormone receptors, the super-family contains a number of proteins that have no known ligands, termed orphan nuclear hormone receptors (Mangelsdorf et al, Cell 83:835-839 (1995), O'Malley et al., Mol. Endocrinol. 10: 1293 (1996), Enmark et al., Mol. Endocrinol. 10, 1293-1307 (1996), and Giguere, Endocrinol. Rev. 20: 689-725 (1999)). The conventional nuclear hormone receptors are generally transactivators in the presence of ligand, and can either be active repressors or transcriptionally inert in the absence of ligand. Some of the orphan receptors behave, as if they are transcriptionally inert in the absence of ligand. Others, however, behave as either constitutive activators of repressors. These orphan nuclear hormone receptors are either under the control of ubiquitous ligands that have not been identified, or do not need to bind ligand to exert these activities. Since the expression of a large number of genes is regulated by the NHR's and since NHR's are expressed in many cell types, modulation of NHR's through binding of either natural hormones or synthetic NHR's ligands can have profound effects on the physiology and pathophysiology of the organism.

In common with other transcription factors, the nuclear hormone receptors have a modular structure, being comprised of three distinct domains: an N-terminal domain of variable size containing a transcriptional activation function AF-1, a highly conserved DNA binding domain and a moderately conserved ligand-binding domain. The ligand- binding domain is not only responsible for binding the specific ligand but also contains a transcriptional activation function called AF-2 and a dimerization domain (Wurtz et al., Nature Struc. Bio. 3:87-94 (1996), Parker et al, Nature Struc. Biol, 3:113-115 (1996) and Dumar et al., Steroids 64:310-319 (1999)). Although the overall protein sequence of these receptors can vary significantly, all share both a common structural arrangement indicative of divergence from an ancestral archetype, and substantial homology (especially sequence identity) at the ligand-binding domain.

The steroid binding nuclear hormone receptors (SB-NHR's) comprise a sub-family of nuclear hormone receptors. These receptors are related in that they share a stronger sequence homology to on another, particularly in the ligand-binding domain (LDB), than to the other members of the NHR super-family (Evans, 1988, vide supra) and they all utilize steroid based ligands. Some examples of this sub-family of NHR's are the androgen receptor (AR), the estrogen receptor (ER), the progesterone receptor (PR), the glucocorticoid receptor (GR), the mineralocorticoid receptor (MR), the aldosterone receptor (ALDR) and the steroid and xenobiotic receptor (SXR) (Evans et al., WO 99/35246). Based on the strong sequence homology in the LBD, several orphan receptors (e.g., the liver X receptor, LXR) may also be members of the SB-NHR subfamily. Consistent with the high sequence homology found in the LBD for each of the SB- NHR's, the natural ligands for each is derived from a common steroid core. Examples of some of the steroid based ligands utilized by members of the SB-NHR's include cortisol, aldosterone, estrogen, progesterone, testosterone, and dihydrotestosterone. Specificity of a particular steroid based ligand for one SB-NHR versus another is obtained by differential substitution about the steroid core. High affinity binding to a particular SB-NHR, coupled with high level specificity for that particular SB-NHR, can be achieved with only minor structural changes about the steroid core (e.g., Waller et al., Toxicol. Appl. Pharmacol. 137:219-227 (1996) andMekenyan et al. Environ. Sci. Tecnol. 31:3702-3711 (1997)), binding affinity for progesterone towards the androgen receptor as compared to testosterone).

Numerous synthetically derived steroidal and non-steroidal agonists and antagonists have been described for the members of the SB-NHR family. Many of these agonist and antagonist ligands are used clinically in man to treat a variety of medical conditions. RU-486 is an example of an synthetic agonist of the PR, which is utilized as a birth control agent (Negeto et al., Cell 69:703-713 (1992)), and flutamide is an example of an antagonist of the AR, which is utilized for the treatment of prostate cancer (Νeri et al, Endocrinol. 91:427-437 (1972)). Tamoxifen is an example of a tissue specific modulator of the ER function, that is used in the treatment of breast cancer (Smigel, J. Natl. Cancer Inst., 90:647-648 (1998)). Tamoxifen can function as an antagonist of the ER in breast tissue while acting as an agonist of the ER in bone (Grese et al., Proc. Natl. Acad. Sci. USA 94:14105-14110 (1997)). Because of the tissue selective effects seen for tamoxifen, this agent and agents like it are referred to as " mixed agonist/antagonist". In addition to synthetically derived non-endogenous ligands, non- endogenous ligands for ΝHR's can be obtained from food source (Regal et al., Proc. Soc. Exp. Biol. Med. 223:372-378 (2000) and Hempstock et al., J. Med. Food 2:267-269 (1999)). The ability to modulate the transcriptional activity of individual ΝHR by the addition of a small molecule ligand, makes them ideal targets for the development of pharmaceutical agents for a variety of disease states. As mentioned above, non-natural ligands can be synthetically engineered to serve as modulators of the function of NHR's. In the case of SB-NHR's, engineering of an unnatural ligand can include the identification of a core structure which mimics the natural steroid core system. This can be achieved by random screening against several SB-NHR's or through directed approaches using the available crystal structures of a variety of NHR ligand binding domains (Bourguet et al, Nature 375:377-382 (1995), Brzozowski, et al., Nature, 389, 753-758 (1997), Shiau et al., Cell 95:927-937 (1998) and Tanenbaum et al., Proc. Natl. Acad. Sci. USA 95:5998-6003 (1998). Differential substitution about such a steroid mimic core can provide agents with selectivity for one receptor versus another. In addition, such modifications can be employed to obtain agents with agonist or antagonist activity for a particular SB-NHR. Differential substitution about the steroid mimic core can result in the formation of a series of high affinity agonists and antagonists with specificity for, for example, ER versus PR versus AR versus GR versus MR. Such an approach of differential substitution has been reported, for example for quinoline based modulators of steroid NHR in J. Med. Chem. 41:623 (1999); WO 97/49709; US 5696133; US 5696130; US 5695693647; US 5693646; US 5688810; US 5688808 and WO 96/19458, all incorporated herein by reference.

There is a need for compounds that can produce the same positive responses as the natural NHR's ligands without the negative side effects. Also needed are NHR's ligands that exert selective effects on different tissues of the body.

The compounds of the instant invention comprise a core which serves as a steroid mimic, and are useful as modulators of the function of steroid binding nuclear hormone receptors, as well as other NHR as described in the following and as such may be useful for treatment or prevention of a variety of conditions related to NHR functioning including cardiovascular, metabolic, and CNS diseases, immune response, and cancer. Two prior art compounds, 5,6,7,8,9,10-hexahydro-benzo[/]quinolin-3(4H)-one (1) 3- ethoxy-5,6,7,8,9,10-hexahydro-benzo[rlquinohne (2), that are related to the instant invention are described in Danishefsky and Feldman, Tetrahedron 1968 24(ll):4083-9. However neither of these substances have significant affinity for any of the nuclear hormone receptors that we have screened against (IC₅₀ > 50 μM for AR, ER-α, ER-β, GR, MR, PR, PPAR-α, PPAR-γ, PPAR-δ). Therefore the compounds of the instant invention have unexpected affinity for nuclear hormone receptors.

DESCRIPTION OF INVENTION In accordance with the present invention, compounds are provided which are nuclear hormone receptor ligands and have the general formula I or II:

wherein R₇α, R₇β, R₈ , R₈β, R₉ , R₉β, R,₀α, and R_ι0β, (of compounds of general formula I or II) are the same or are different and each is an R^A group and at least one substituent is not a hydrogen atom; R^Λ is selected from the group of: hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, carboxyalkyl, carboxylate, and cyano;

R₃ (of compounds of general formula I) is selected from: a hydrogen atom, a linear or branched alkyl or cycloalkyl group, and acyl group of 1 to 4 carbon atoms;

R₄ (of compounds of general formula II) is a hydrogen atom, a linear or branched alkyl or cycloalkyl group, and acyl group of 1 to 4 carbon atoms.

DETAILED DESCRIPTION OF INVENTION

The present invention relates to compounds useful as nuclear hormone receptor modulators and have the general formula I or II as described above.

Preferably, the compounds of formula I or II are compounds wherein R₉α, R₉β, Rι₀α, and Rioβ are hydrogen atoms. Particularly preferred compounds of the general formula II are compounds wherein R is R^B and R^B is selected from the group: hydrogen, methyl, ethyl, n-propyl, z-propyl, 2-propenyl, 2-propynyl, n-butyl, cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl, benzyl, and phenethyl.

Compounds of the invention include, but are not limited to, the following:

(rαc)-3-Oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ lquinoline-8-carboxylic acid methyl ester

(Ela);

(ra^-S-Oxo-S^^^J.δ^jlO-octahydro-benzol lquinoline^-carboxylic acid methyl ester

(Elb);

(ra^^-Methyl-S-oxo-S^^jό .δ^JO-octahydro-benzol Iquinoline-S-carboxylic acid methyl ester (E2a); (r c)-4-Methyl-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[/]quinoline-7-carboxylic acid methyl ester (E2b);

(rαc)-4-(2-Methyl-benzyl)-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[f]quinoline-8- carboxylic acid (E3a); and

(rαc)-4-(2-Methyl-benzyl)-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[fJquinoline-7- carboxylic acid (E3b).

In one embodiment of the invention there is provided a method of eliciting an nuclear hormone receptor modulating effect in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of any of the compounds or any of the pharmaceutical compositions described above.

In a first class of this embodiment is the method wherein the nuclear hormone receptor modulating effect is an agonizing effect.

A subclass of this class is the method wherein the nuclear hormone receptor is an androgen receptor.

A second subclass of this class is the method wherein the nuclear hormone receptor is an estrogen receptor.

A third subclass of this class is the method wherein the nuclear hormone receptor is a glucocorticoid receptor.

A fourth subclass of this class is the method wherein the nuclear hormone receptor is a liver X receptor.

A fifth subclass of this class is the method wherein the nuclear hormone receptor is a mineralocorticoid receptor. A sixth subclass of this class is the method wherein the nuclear hormone receptor is a progesterone receptor.

A seventh subclass of this class is the method wherein the nuclear hormone receptor is peroxisome proliferator-activated receptor.

In a second class of this embodiment there is provided the method wherein the nuclear hormone receptor modulating effect is an antagonizing effect.

A fifth subclass of this class is the method wherein the nuclear hormone receptor is a mineralocorticoid receptor.

A sixth subclass of this class is the method wherein the nuclear hormone receptor is a progesterone receptor.

A seventh subclass of this class is the method wherein the nuclear hormone receptor is peroxisome proliferator-activated receptor. Yet another embodiment of the invention is a method of treating or preventing metabolic or autoimmune diseases, or cancer in a mammal in need thereof by administering to the mammal a therapeutically effective amount of any of the compounds or pharmaceutical compositions described above and below.

Exemplifying the invention is a pharmaceutical composition comprising any of the compounds described above and a pharmaceutically acceptable carrier. Also exemplifying the invention is a pharmaceutical composition made by combining any of the compounds described above and a pharmaceutically acceptable carrier. An illustration of the invention is a process for making a pharmaceutical composition comprising combining any of the compounds described above and a pharmaceutically acceptable carrier.

Further exemplifying the invention is a method of treating or preventing proliferative diseases, cancers, benign prostate hyperplasia, adenomas and neoplasisms of the prostate, benign or malignant tumor cells containing the androgen receptor, cardiovascular diseases, angiogenic conditions or disorders, hirsutism, acne, hyperpilosity, inflammation, immune modulation, seborrhea, endometriosis, polycycstic ovary syndrome, androgen alopecia, hypogonadism, osteoporosis, suppressing spermatogenesis, libido, cachexia, anorexia, cancers expressing the estrogen receptor, prostate cancer, breast cancer, endometrial cancer, incontinence, hot flashes, vaginal dryness, menopause, contraception, pregnancy termination, cancers containing the progesterone receptor, endometriosis, cachexia, menopause fibrosis, labor induction, autoimmune diseases, impairment of cognitive functioning, cerebral degenerative disorders, Alzheimer's disease, psychotic disorders, type-II diabetes, congestive heart failure, disregulation of cholesterol or lipid homeostasis, and/or obesity in a mammal in need thereof by administering to the mammal a therapeutically effective amount of any of the compounds described above. Further exemplifying the invention is the use of any of the compounds described above in the preparation of a medicament for the treatment and/or prevention of conditions or disorders selected from the group consisting of proliferative diseases, cancers, benign prostate hyperplasia, adenomas and neoplasisms of the prostate, benign or malignant tumor cells containing the androgen receptor, cardiovascular diseases, angiogenic conditions or disorders, hirsutism, acne, hyperpilosity, inflammation, immune modulation, seborrhea, endometriosis, polycycstic ovary syndrome, androgen alopecia, hypogonadism, osteoporosis, suppressing spermatogenesis, libido, cachexia, anorexia, cancers expressing the estrogen receptor, prostate cancer, breast cancer, endometrial cancer, incontinence, hot flashes, vaginal dryness, menopause, contraception, pregnancy termination, cancers containing the progesterone receptor, endometriosis, cachexia, menopause fibrosis, labor induction, autoimmune diseases, impairment of cognitive functioning, cerebral degenerative disorders, Alzheimer's disease, psychotic disorders, type-II diabetes, congestive heart failure, disregulation of cholesterol or lipid homeostasis, and/or obesity.

The compounds of the present invention can be used in combination with other agents useful for treating nuclear hormone receptor-mediated conditions. The individual components of such combinations can be administer separately at different times during the course of therapy or concurrently in divided or single combination forms. The instant invention is therefore to be understood as embracing all such regimes of simultaneous or alternating treatment and the term "administering" is to be interpreted accordingly. It will be understood that the scope of combinations of the compounds of this invention with other agents useful for treating estrogen-mediated conditions includes in principle any combination with any pharmaceutical composition useful for treating disorders related to estrogen functioning.

The compounds of the present invention can be administered in such oral dosage forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powder, granules, elixirs, tinctures, suspensions, syrups and emulsions. Likewise, they may also be administered in intravenous (bolus or infusion), intraperitoneal, topical (e.g., ocular eyedrop), subcutaneous, intramuscular, or transdermal (e.g., patch) form, all using forms well known to those of ordinary skill in the pharmaceutical arts.

The dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex, and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician, veterinarian or clinician can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition.

Oral dosages of the present invention, when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 100 mg/kg/day, preferably 0.01 mg per kg of body weight per day (mg/kg/day) to 10 mg/kg/day, and most preferably 0.1 to 5.0 mg/kg/day. For oral administration, the compositions are preferably provided in the form of tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, and 500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from about 1 mg to about 100 mg of active ingredient. Intravenously, the most preferred doses will range from about 0.1 to about 10 mg/kg/minute during a constant rate infusion. Advantageously, compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, preferred compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches will known to those of ordinary skill in the art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.

In the methods of the present invention, the compounds herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, exipients or carriers (collectively referred to herein as "carrier" materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.

For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, com sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms includes sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include without limitation starch, methylcellulose, agar, bentonite, xanthan gum and the like.

The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed form a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines. The following definitions apply to the terms as used throughout this specification, unless otherwise limited in specific instances.

The term "nuclear hormone receptor ligand" as used herein is intended to cover any moiety which binds to a nuclear hormone receptor. The ligand may act as an agonist, an antagonist, a partial agonist or a partial antagonist. The ligand may be either selective for a particular nuclear hormone receptor or display mixed nuclear hormone receptor activity.

The term "aliphatic hydrocarbon(s)" as used herein refers to acyclic straight or branched chain groups which include alkyl, alkenyl or alkynyl groups.

The term "aromatic hydrocarbon(s)" as used herein refers to groups including aryl groups as defined herein.

Unless otherwise indicated, the term "lower alkyl", "alkyl", or "alk" as employed herein alone or as part of another group includes both straight and branched chain hydrocarbons, containing 1 to 12 carbon atoms (in the case of alkyl) in the normal chain and preferably 1 to 6 carbons, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, or isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4- trimethylpentyl, nonyl, decyl, undecyl, dodecyl.

The term "cycloalkyl" as employed herein alone or as part of another group refers to 3- to 7-membered fully saturated monocyclic ring system and include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

The term "cycloalkylalkyl" as employed herein alone or as part of another group refers to an cycloalkyl group containing 3 to 7 carbon atoms attached through available carbon atoms to a straight or branched chain alkyl radical containing 1 to 6 carbon atoms and include but are not limited to cyclopropylmethyl (-CH₂C₃Hs), cyclobutylethyl (- CH₂CH₂C4H₇), and cyclopentylpropyl (-CH₂CH₂CH₂C₅H₉).

The term "aryl" as employed herein alone or as part of another group refers to monocyclic and bicyclic aromatic groups containing 6 to 10 carbons in the ring portion and include but are not limited to phenyl, 1-naphthyl, and 2-naphthyl and may be optionally substituted through available carbon atoms with 1, 2, or 3 groups selected from hydrogen, halo, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, amino, trifluoromethyl, trifluoromethoxy, alkynyl, hydroxy, nitro, cyano, or carboxy.

The term "arylalkyl" as employed herein alone or as part of another group refers to an aryl group containing 6 to 10 carbon atoms attached through available carbon atoms to a straight or branched chain alkyl radical containing 1 to 6 carbon atoms and include but are not limited to benzyl (-CH₂Ph), phenethyl (-CH CH Ph), phenpropyl (- CH₂CH₂CH₂Ph), and 1-napthylmethylene (-CH₂C₁₀H₇).

Unless otherwise indicated, the term "lower alkenyl" or "alkenyl" as used herein by itself or as part of another group refers to straight or branched chain radicals of 2 to 12 carbons, preferably 2 to 6 carbons, in the normal chain, which include one to six double bonds in the normal chain, such as vinyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 3-octenyl, 3- nonenyl, 4-decenyl, 3-undecenyl, 4-dodecenyl, and the like.

Unless otherwise indicated, the term "lower alkynyl" or "alkynyl" as used herein by itself or as part of another group refers to straight or branched chain radicals of 2 to 12 carbons, preferably 2 to 6 carbons, in the normal chain, which include one triple bond in the normal chain, such as 2-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, 2- hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonynyl, 4- decynyl, 3-undecynyl, 4-dodecynyl and the like. The term "halogen" or "halo" as used herein alone or as part of another group refers to chlorine, bromine, fluorine, and iodine as well as CF₃.

The term "acyl" as employed herein alone or as part of another group refers to a carbonyl group (C=O) which in turn is bonded a linear or branched alkyl group of from 1 to 4 carbon atoms and includes but is not limited to acetyl [-(C=O)CH₃], propionyl [- (C=O)CH₂CH₃], and butyryl KC=O)CH₂CH₂CH₃].

The compounds of formula I or II can be present as salts, in particular pharmaceutically acceptable salts. If the compounds of formula I or II have, for example, at least one basic center, they can form acid addition salts. These are formed, for example, with strong inorganic acids, such as mineral acids, for example sulfuric acid, phosphoric acid or a hydrohalic acid, with strong organic carboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atoms which are unsubstituted or substituted, for example, by halogen, for example acetic acid, such as saturated or unsaturated dicarboxylic acids, for example oxalic, malonic, succinic, maleic, fumaric, phthalic or terephthalic acid, such as hydroxycarboxylic acids, for example ascorbic, glycolic, lactic, malic, tartaric or citric acid, such as amino acids, (for example aspartic or glutamic acid or lysine or arginine), or benzoic acid, or with organic sulfonic acids, such as (C_j-C₄)-alkyl- or aryl-sulfonic acids which are unsubstituted or substituted, for example by halogen, for example methane- or p-toluene-sulfonic acid. Corresponding acid addition salts can also be formed having, if desired, an additionally present basic center. The compounds of formula I or II having at least one acid group (for example COOH) can also form salts with bases. Suitable salts with bases are, for example, metal salts, such as alkali metal or alkaline earth metal salts, for example sodium, potassium or magnesium salts, or salts with ammonia or an organic amine, such as morpholine, thiomorpholine, piperidine, pyrrolidine, a mono-, di- or tri-lower alkylamine, for example ethyl-, tert-butyl-, diethyl-, diisopropyl-, triethyl-, tributyl- or dimethyl-propylamine, or a mono-, di- or trihydroxy lower alkylamine, for example mono-, di- or triethanolamine. Corresponding internal salts may furthermore be formed. Salts which are unsuitable for pharmaceutical uses but which can be employed, for example, for the isolation or purification of free compounds I or their pharmaceutically acceptable salts are also included.

Preferred salts of the compounds of formula I or II which include a basic group include monohydrochloride, hydrogensulfate, methanesulfonate, phosphate or nitrate.

Preferred salts of the compounds of formula I or II which include an acid group include sodium, potassium and magnesium salts and pharmaceutically acceptable organic amines.

The compounds in the invention contain at least one chiral center and therefore exist as optical isomers. The invention therefore comprises the optically inactive racemic (rac) mixtures (a one to one mixture of enantiomers), optically enriched scalemic mixtures as well as the optically pure individual enantiomers. The compounds in the invention also may contain more than one chiral center and therefore may exist as diastereomers. The invention therefore comprises individual diastereomers as well as mixtures of diastereomers in cases where the compound contains more than one stereo center. The compounds in the invention also may contain acyclic alkenes or oximes and therefore exist as either the E (entgegen) or Z (zusammen) isomers. The invention therefore comprises individual E or Z isomers as well as mixtures of E and Z isomers in cases where the compound contains an acylic alkene or oxime funtional group. Also included within the scope of the invention are polymorphs, hydrates, and solvates of the compounds of the instant invention.

The present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of the compounds of this invention which are readily convertible in vivo into the required compound. Thus, in the methods of treatment of the present invention, the term "administering" shall encompass the treatment of the various conditions described with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the patient. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example in "Design of Prodrugs" ed. H. Bundgaard, Elsevier, 1985, which is incorporated by reference herein in its entirety. Metabolites of the compounds includes active species produced upon introduction of compounds of this invention into the biological milieu.

The present invention also relates to pharmaceutical compositions comprising the compounds of the present invention and a pharmaceutically acceptable carrier.

The present invention also relates to methods for making the pharmaceutical compositions of the present invention.

The present invention also relates to methods for eliciting a nuclear hormone receptor modulating effect in a mammal in need thereof by administration of the compounds and pharmaceutical compositions of the present invention.

The present invention also relates to methods for eliciting a nuclear hormone receptor antagonizing effect in a mammal in need thereof by administration of the compounds and pharmaceutical compositions of the present invention.

The present invention also relates to methods for treating or preventing disorders elated to nuclear receptor functioning, proliferative diseases, cancers, benign prostate hyperplasia, adenomas and neoplasisms of the prostate, benign or malignant tumor cells containing the androgen receptor, cardiovascular diseases, angiogenic conditions or disorders, hirsutism, acne, hyperpilosity, inflammation, immune modulation, seborrhea, endometriosis, polycycstic ovary syndrome, androgen alopecia, hypogonadism, osteoporosis, suppressing spermatogenesis, libido, cachexia, anorexia, cancers expressing the estrogen receptor, prostate cancer, breast cancer, endometrial cancer, incontinence, hot flashes, vaginal dryness, menopause, contraception, pregnancy termination, cancers containing the progesterone receptor, endometriosis, cachexia, menopause fibrosis, labor induction, autoimmune diseases, impairment of cognitive functioning, cerebral degenerative disorders, Alzheimer's disease, psychotic disorders, type-II diabetes, congestive heart failure, disregulation of cholesterol or lipid homeostasis, and/or obesity.

The novel compounds of the present invention can be prepared according to the procedure of the following Schemes and examples, using appropriate materials and are further exemplified by the following specific examples. The compounds illustrated in the examples are not, however, to be construed as forming the only genus that is considered as the invention. The following examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variation of the conditions and processes of the following preparative procedures can be used to prepare these compounds. The compounds of the present invention are prepared according to the general methods outlined in Schemes 1- 2, and according to the methods described. All temperatures are degrees Celsius unless otherwise noted. The following abbreviations, reagents, expressions or equipment, which are amongst those used in the descriptions below, are explained as follows: 20- 25°C (room temperature, r.t.), molar equivalent (eq.), dimethyl formamide, (DMF) dichloromethane (DCM), ethyl acetate (EtOAc), tetrahydrofuran (THF), lithium diisopropylamide (LDA), methyl t-butyl ether (MTBE), rotating glass sheet coated with a silica gel-gypsum mixture used for chromatographic purification (chromatotron), preparative liquid chromatography with a C8 stationary phase and ammonium acetate acetonitrile-water buffer as mobile phase (PHPLC), electrospray mass spectroscopy (ES/MS).

A general route for the construction of the 3-oxo-3 ,4,5,6,7,8,9, 10-octahydro- benzo[ ]quinoline nucleus is shown in Scheme 1. This methodology is based on the chemistry described by Ruda, et al., /. Comb. Chem 2002, 4, 530-535 and Dubas- Sluyter, et al., Rec. Trav. Chim. 1972, 91, 157-160. In step 1, an alkyl propiolate and 3- amino-2-cyclohexen-l-one are coupled in refmxing DMF to give 7,8-dihydro-iH,6H- quinoline-2,5-dione (1). Pyridone 1 is then selectively O-alkylated with an alkyl halide to give the 2-alkoxypyridine 2. Treatment of the ketone 2 is then treated with a vinyl organometalic reagent followed by dehydration to afford the diene 3. This diene is then heated with a dieneophile to provide the 3-alkoxy-5,6,6a,7,8,9-hexahydro- benzo[flquinoline (4). Treatment of the Diels-Alder adduct 4 with a alkyl halide results in simultaneous deprotection of the pyridone and migration of the double bond to afford the thermodynamically more stable 3-alkoxy-5,6,6a,7,8,9-hexahydrobenzo[/]quinoline (5). Alternatively treatment of adduct 4 with trimethyl silyl iodide yields the 3-oxo- 3,4,5,6,7,8,9, 10-octahydro-benzo[ ]quinoline (6). The pyridone 6 may be selectively O- alkylated with an alkyl halide to give the 2-alkoxypyridine 7.

Scheme 1: A general solution phase route to 3-alkoxy-5,6,6a,7,8,9- hexahydrobenzo[ ]quinoline (7) and 3-oxo-3 ,4,5,6,7,8,9, 10-octahydro-benzo[^|quinoline (5 or 6) derivatives.

Reaction conditions: (a) DMF/Δ; (b) alkyl halide/Ag₂CO₃/toluene; (c) vinyl magnesium bromide/THF; (d) 2 M HCl/THF; (e) dieneophile/toluene/Δ; (f) alkyl halide DMF/Δ; (g) TMSCl/Nal/DCM/Δ.

Scheme 2: A general solid phase route to 3-alkoxy-5,6,6a,7,8,9- hexahydrobenzo[/]quinoline (7) and 3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[/]quinoline (5 or 6) derivatives.

A general route for the construction of the 3-oxo-3,4,5,6,7,8,9,10-octahydro- benzo[/]quinoline nucleus employing solid phase chemistry is shown in Scheme 2. This methodology is based on the chemistry described by Ruda, et al., J. Comb. Chem 2002, 4, 530-535. Pyridone 1 is selectively linked through the oxygen atom to a solid phase resin under Mitsunobu conditions (see Chen and Munoz Tetrahedron Lett. 1998, 39, 6781-6784; Comins and Gao Tetrahedron Lett. 1994, 35, 2819-2822) to give the 2- alkoxypyridine 8. Treatment of the ketone 8 is then treated with a vinyl organometalic reagent followed by dehydration to afford the diene 9. This diene is then heated with a dieneophile to provide the 3-alkoxy-5,6,6a,7,8,9-hexahydro-benzo[/]quinoline (10). Treatment of the Diels-Alder adduct 10 with trimethylsilyl iodide results in simultaneous deprotection of the pyridone and migration of the double bond to afford the thermodynamically more stable 3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline (5).

Reaction conditions: (a) Ph₃P DEAD/Δ; (b) alkyl halide/Ag₂CO₃/toluene; (c) vinyl magnesium bromide/THF; (d) 2 M HCl THF; (e) dieneophile/toluene/Δ; (f) alkyl halide DMF/Δ; (g) TMSCl/Nal/DCM/Δ.

EXAMPLES

The following Examples represent preferred embodiments of the present invention. However, they should not be construed as limiting the invention in any way.

Example 1: Synthesis of 3-Oxo-3,4.5.6 .8.9.10-octahvdro-benzorι1quinoline-8- carboxylic acid methyl ester (Εla) and 3-Oxo-3 A5.6-7,8,9-10-octahvdro- benzorflquinoline-7-carboxylic acid methyl ester (Elb).

Step 1: 7-8-Dihydro-lH,6H-quinoline-2,5-dione.

Methyl propiolate (2.69 g, 32 mmol) and 3-amino-2-cyclohexen-l-one (3.5 g, 32 mmol) were dissolved in DMF (20 mL) and heated to reflux. After 3 hrs the mixture was cooled on an ice bath, and the crystalline material was collected and washed with a small amount of DMF cooled in dry ice. The filtrate was heated again for 16 hr after addition of methyl propiolate (2.69 g, 32 mmol) and the solid precipitate was collected and washed with a small amount of DMF. This procedure is repeated once again. The crude solid was recrystallized from methanol and to yield 1.93 g (37 %) of the desired 7,8-dihydro-lH,6H-quinoline-2,5-dione. 1H NMR (270 MHz, CDC1₃) δ 8.04 (d, 1H, J = 9.5 Hz), 6.45 (d, 1H, J = 9.5 Hz), 2.94 (t, 2H, J = 6.2 Hz), 2.57 (t, 2H, J = 6.2 Hz) and 2.12-2.21 (m, 2H); ¹³C NMR (67 MHz, CDC1₃) δ 193.8, 165.9, 156.1, 139.1, 118.1, 115.0, 37.2, 27.3 and 21.4; ES-MS mlz 164 ((M+H)⁺).

Step 2: 2-Benzyloxy-7,8-dihydro-6H-quinoIin-5-one.

A mixture of 7,8-Dihydro-lH,6H-quinoline-2,5-dione (1.138 g, 6.97 mmol), silver carbonate (1.138 g, 4.13 mmol) and benzyl bromide (1 mL) in toluene (50 mL) was stirred for 3 days in the dark at room temperature. The reaction mixture was filtered through Celite^®, and the filtrate was evaporated to leave an orange oil, which was loaded on a silica gel column. Excess unreacted benzyl bromide was first removed by eluting with n-heptane and then the product was eluted with 2:8 ethyl acetate:heptane to yield 1.64 g (93 %) of the desired 2-benzyloxy-7,8-dihydro-6H-quinolin-5-one. 1H NMR (270 MHz, CDC1₃) δ 8.05 (d, 1H, J = 8.6 Hz), 7.19-7.40 (m, 5H), 6.58 (d, 1H, J = 8.6 Hz), 5.35 (s, 2H), 2.90 (t, 2H, J = 6.2 Hz), 2.48 (t, 2H, J = 6.0 and 7.2 Hz) and 1.94- 2.04 (m, 1H); ¹³C NMR (67 MHz, CDC1₃) δ 196.7, 165.5, 163.8, 137.8, 136.8, 128.5, 128.2, 128.1, 123.0, 110.2, 68.2, 38.2, 32.5 and 22.0; ES-MS mlz 254 ((M+H)⁺).

Step 3: 2-Benzyloxy-5-vinyl-7,8-dihydro-quinoline.

A solution of 2-benzyloxy-7,8-dihydro-6H-quinolin-5-one (116 mg, 0.46 mmol) in dry THF (2 mL) was added slowly to a solution of vinyl magnesium bromide (4 mL, 1 M) in dry THF (10 mL) at 0 ^°C. After 2 hrs the reaction was poured on ice and 2M HCl (20 mL) was added and the reaction was stirred over night. The reaction mixture was then neutralized with aqueous sodium bicarbonate and extracted with ethyl acetate. The organic phase was dried with anhydrous sodium sulfate, evaporated and chromatographed on a silica gel column eluted with 1:9 ethyl acetate:heptane to yield 73.8 mg (61 %) of the desired 2-benzyloxy-5-vinyl-7,8-dihydro-quinoline. 1H NMR (270 MHz, CDC1₃) δ 7.54 (d, IH, J= 8.4 Hz), 7.30-7.50 (m, 5H), 6.62 (d, IH, J = 8.4 Hz), 6.53 (dd, IH, J = 10.9 and 17.3 Hz), 6.06 (t, IH, J = 4.8 Hz), 5.49 (d, IH, J = 17.3 Hz), 5.39 (s, 2H), 5.19 (d, IH, J = 10.9 Hz), 2.87 (t, 2H, J = 8.3) and 2.36-2.44 (m, 2H); ¹³C NMR (67 MHz, CDC1₃) δ 162.0, 155.6, 137.6, 135.2, 134.9, 134.1, 128.5, 128.1, 127.8, 124.3, 123.4, 115.4, 107.8, 67.7, 30.7 and 23.1; ES-MS mlz 269 ((M+H)⁺).

Step 4: 3-Benzyloxy-5,6,6a,7,8,9-hexahydro-benzo[/]quinoline-8-carboxylic acid methyl ester and 3-Benzyloxy-5,6,6a,7,8,9-hexahydro-benzo| lquinoline-7-carboxylic acid methyl ester.

A solution of 2-benzyloxy-5-vinyl-7,8-dihydro-quinoline (73 mg, 0.28 mmol) and of methyl acrylate (2 mL) in toluene (5 mL) was added to a sealed reaction tube and heated to 120 C. The reaction was completed after 1.5 hrs as judged by TLC. The reaction mixture was evaporated and the crude was purified on a silica column 1:9 ethyl acetate: heptane to afford 61 mgs (62%) of the desired 3-benzyloxy-5,6,6a,7,8,9- hexahydro-benzo[ ]quinoline-8-carboxylic acid methyl ester and 3-benzyloxy- 5,6,6a,7,8,9-hexahydro-benzo[ ]quinoline-7-carboxylic acid methyl ester as a 1:1 mixture. ES-MS mlz 350 ((M+H)⁺).

Step 5: 3-Oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-8-carboxylic acid methyl ester (Ela) and 3-Oxo-3,4,5,6,7,8,9,10-octahydro-benzo[/]quinoline-7-carboxylic acid methyl ester (Elb).

A solution of a 1:1 mixture of 3-benzyloxy-5,6,6a,7,8,9-hexahydro-benzo|/|quinoline-8- carboxylic acid methyl ester and 3-benzyloxy-5,6,6a,7,8,9-hexahydro- benzo[ ]quinoline-7-carboxylic acid methyl ester (37 mg 0.11 mmol), trimethylsilyl chloride (46 mg, 0.42 mmol) and a large excess of sodium iodide (0.5 mL) in DCM (5 mL) were heated to 60 C in a sealed reaction tube. The reaction was followed with TLC. The reaction was quenched with MeOH after TLC (-1 hr) showed that all the starting material had been consumed. The reaction mixture was evaporated and the residue was extracted with DCM and water. The organic phase was evaporated and purified on a silica column eluted (95%:5%) DCM:MeOH to afford 27 mgs (95%) of 1:1 mixture of the desired 3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[f]quinoline-8- carboxylic acid methyl ester (Ela) and 3-oxo-3,4,5,6,7,8,9,10-octahydro- benzo[f]quinoline-7-carboxylic acid methyl ester (Elb). ES-MS mlz 260 ((M+H)⁺).

Example 2; Synthesis of 4-Methyl-3-oxo-3,4,5.6 ,8,9,10-octahydro-benzort1quinoline- 8-carboxylic acid methyl ester (E2a and 4-Methyl-3-oxo-3,4.5,6,7,8,9.10-octahvdro- benzorrlquinoline-7-carboxylic acid methyl ester (E2b).

A 1:1 mixture of 3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-8-carboxylic acid methyl ester (Ela) and 3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-7- carboxylic acid methyl ester (Elb) (10 mg, 0.04 mmol) prepared as described in Example 1 were mixed with a large excess of lithium carbonate and a large excess of methyl iodide (100 μL) in DMF (1 mL) and then heated to 80 C. When TLC showed that all the starting material had been consumed after 8 hrs and the reaction was stopped. The product was purified by filtration trough dry silica plug in a syringe and eluted with (95%: 5%) DCM:MeOH to yield 10.9 mgs (100%) of a 1 : 1 mixture of the desired 4-methyl-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[/]quinoline-8-carboxylic acid methyl ester (6a) and 4-methyl-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-7- carboxylic acid methyl ester (6b). ES-MS mlz 21 ((M+H)⁺).

Example 3: Synthesis of 2'-methyl-4-benzyl-3-oxo-3A5,6.7,8,9,10-octahydro- benzorflquinoline-8-carboxylic acid methyl ester (E3a) and 2'-methyl-4-benzyl-3-oxo- 3A5.6 ,8,9,10-octahvdro-benzorflquinoline-7-carboxylic acid methyl ester (E3b).

A 1:1 mixture of 3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[/]quinoline-8-carboxylic acid methyl ester (Ela) and 3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-7- carboxylic acid methyl ester (Elb) (7.1 mg, 0.027 mmol) prepared as described in Example 1 were mixed with lithium carbonate (4 mg, 0.055 mmol) and 2-methyl benzyl bromide (20 mg, 0.11 mmol) in DMF (1 mL) and then heated to 100 C. When TLC showed that all the starting material had been consumed after 8 hrs and the reaction was stopped. The solvent was evaporated and the residue was dissolved in a solution of THF (1 mL) and lithium hydroxide (0.5 mL, 1 M). When TLC showed that all the starting material had been consumed the reaction was extracted with DCM and then purified by filtration trough dry silica plug in a syringe and eluted with (95%:5%) DCM: MeOH to yield 9.8 mgs (100%) of a 1:1 mixture of the desired 2'-methyl-4- benzyl-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-8-carboxylic acid methyl ester (E3a) and 2'-methyl-4-benzyl-3-oxo-3,4,5,6,7,8,9,10-octahydro- benzo|]flquinoline-7-carboxylic acid methyl ester (E3b). ES-MS mlz 350 ((M+H)⁺).

DESCRIPTION OF THE SCINTISTRIP ER BINDING ASSAY Introduction

The scintistrip assay differs from a traditional hormone binding assay by not requiring the removal of free tracer prior to the measurement of receptor bound tracer. The scintillating agent is in the polystyrene forming the incubation vial and thus a radioactive molecule in the close proximity to the surface will induce scintillation of the plastic. For ³[H]-labeled ligands, the distance between the free tracer and the scintillating polystyrene surface is too far to induce scintillation of the plastic while ³[H]-labeled ligands bound to receptors immobilized on the surface are close enough to induce scintillation¹ thus enabling a convenient way to measure the competition between a non-radioactive estrogen receptor interacting agent (the compound to be tested) and a fixed concentration of tracer (³[H]-Estradiol).

Materials and Methods

³[H]-β-Estradiol (NET 317) hereafter referred to as ³[H]-E2 was purchased from New England Nuclear, Boston, MA. The scintistrip wells (1450-419) and the scintillation counters (Microbeta™ 1450-Plus and 1450-Trilux) were all from Wallac, Turku, Finland. Human estrogen receptors (hER) alpha and beta were extracted from the nuclei from SF9-cells infected with a recombinant baculovirus transfer vector containing the cloned hER genes. Recombinant baculovirus was generated utilizing the BAC-TO-BAC expression system (Life Technonlogies) in accordance to instruction from the supplier. The hER coding sequences were cloned into a baculovirus transfer vector by standard techniques. The recombinant baculoviruses expressing hER were amplified and used to infect SF9 cells. Infected cells were harvested 48 hr post infection. A nuclear fraction was obtained as described in² and the nuclei were extracted with a high-salt buffer (17 mM K₂HPO₄, 3 mM KH₂PO₄, 1 mM MgCl₂, 0.5 mM EDTA, 6 mM MTG, 400 mM KCl, 8.7% Glycerol). The concentration of hER's in the extract was measured as specific [³H]-E2 binding with the 025-assay³ and was determined to contain 400 pmols specific bound [3H]-E2/mL nuclear extract in the case of hER-alpha and 1000 pmols/mL nuclear for hER-beta. The total concentration of proteins (as determined with Bradford Reagent, Bio-Rad according to instructions from manufacturer) in the nuclear extracts were ~ 2 mg mL. The equilibrium binding constant (K,,) for [³H]-E2 to hER in solution was determined to 0.05 nM for hER-alpha and to 0.07 nM for hER-beta with the G25-assay for highly diluted extracts (hER - 0.1 nM). The extracts were aliquoted and stored at -80°C.

The scintistrip assay¹ In brief; the nuclear extracts were diluted (50 fold for hER-alpha and 110 fold for hER-beta) in coating buffer (17 mM K₂HPO₄, 3 mM KH₂PO₄, 40 mM KCl, 6 mM MTG). The diluted extracts were added to Scintistrip wells (200 lJwell) and incubated 18-20 hr. at ambient room temperature (22-25 °C). The estimated final concentration of immobilized hER in all experiments was ~1 nM. All incubations were performed in 17 mM K₂HPO₄, 3 mM KH₂PO₄, 140 mM KCl, 6 mM MTG (buffer A). The wells were washed twice after hER coating with 250 μL buffer prior to addition of the incubation solution. All steps were carried out at ambient room temperature (22-25 °C). Determination of Equilibrium binding constants to immobilized hER.s: Dilutions of ³[H]-E2 in buffer ± Triton X100 were added to the wells (200 μlJwell), the wells were incubated for 3 hr and then measured in the Microbeta. After the measurement an aliquot of the buffer was taken out and counted by regular liquid scintillation counting for determination of the "free" fraction of ³[H]-E2. In order to correct for non-specific binding parallel incubations were done in presence of a 200-fold excess of unlabeled 17-β-E2. The equilibrium dissociation constants (Kj) were calculated as free concentration of ³[H]-E2 at half maximum binding by fitting data to the Hill equation; b = (b_max x Lⁿ)/(Lⁿ+Kdⁿ) where b is specific bound ³[H]-E2, b_max is the maximum binding level, L is the free concentration of [³H]E2, n is the Hill coefficient (the Hill equation equals the Michaelis-Menten equation when n = 1). The equilibrium binding constants were determined to 0.15 - 0.2 nM for both hER subtypes.

Regular competition binding: Samples containing 3 nM [3HQ-E2 plus a range of dilutions of the compounds to be tested were added to wells with immobilized hER and incubated for 18-20 hr at ambient room temperature. The compounds to be tested were diluted in 100% DMSO to a concentration 50 fold higher than the desired final concentration, the final concentration of DMSO was thus 2% in all samples. For compounds able to displace ³[H]-E2 from the receptor an IC₅₀-value (the concentration required to inhibit 50% of the binding of ³[H]-E2) was determined by a non-linear four parameter logistic model; b = ((b_maχ-_bnώ.)/(l+(I/IC_5o)^s))+b„_lin I is added concentration of binding inhibitor, IC₅₀ is the concentration of inhibitor at half maximal binding and S is a slope factor.¹ For determinations of the concentration of ³[H]-E2 in the solutions regular scintillation counting in a Wallac Rackbeta 1214 was performed using the scintillation cocktail Supermix™ (Wallac).

The Microbeta-instrument generates the mean cpm (counts per minute) value / minute and corrects for individual variations between the detectors thus generating corrected cpm values. It was found that the counting efficiency between detectors differed with less than five percent. 1) Haggblad, J., Carlsson, B., Kivela, P., Siitari, H., (1995) Biotechniques 18, 146-151

2) Barkhem, T., Carlsson, B., Simons, J., Moller, B., Berkenstam, A., Gustafsson J.A.G., Nilsson, S. (1991 ) J. Steroid Biochem. Molec. Biol. 38, 667-75

3) Salomonsson, M., Carlsson, B., Haggblad, J., (1994) J. Steroid Biochem. Molec. Biol. 50, 313-318

4) Schultz, J.R., Ruppel, P.I., Johnson, M.A., (1988) in Biopharmaceutical Statistics for Drug Development (Peace, K.E., Ed.) pp. 21-82, Dekker, New York

The compounds of Examples 1-3 exhibit binding affinities to the AR, ER-α, ER-β, GR, MR, PR in the range of IC₅₀ 1 to 100 μM.

Claims

1. A compound having the general formula I or II:

wherein the substituents R₇a, R₇β, R₈ , R₈β, R₉ , R₉β, R₁₀ , and R₁₀β, (of compounds of general formula I or II) are the same or are different and each is an R^A group and at least one substituent is not a hydrogen atom;

R^A is selected from the group of: hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, carboxyalkyl, carboxylate, and cyano;

R₄ (of compounds of general formula II) is selected from: a hydrogen atom, a linear or branched alkyl or cycloalkyl group, and acyl group of 1 to 4 carbon atoms.

2. A compound with the general formula I or II according to claim 1 wherein R₉α, R₉β, R₁₀α, and R^β are hydrogen atoms.

3. A compound with the general formula II according to claim 2 wherein R is R^B and R^B is selected from the group: hydrogen, methyl, ethyl, n-propyl, z-propyl, 2- propenyl, 2-propynyl, rc-butyl, cyclopropylmefhyl, cyclopentylmethyl, cyclohexylmethyl, benzyl, and phenethyl.

4. The compound in claim 1 which is: (r c)-3-Oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-8-carboxylic acid methyl ester (Ela);

(r c)-3-Oxo-3,4,5,6,7,8,9,10-octahydro-benzo[/]quinoline-7-carboxylic acid methyl ester (Elb);

(rac)-4-Methyl-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[/]quinoline-8-carboxylic acid methyl ester (E2a);

(rac)-4-Methyl-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[ ]quinoline-7-carboxylic acid methyl ester (E2b);

(rac)-4-(2-Methyl-benzyl)-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[f]quinoline-8- carboxylic acid (E3a); and

(rac)-4-(2-Methyl-benzyl)-3-oxo-3,4,5,6,7,8,9,10-octahydro-benzo[f]quinoline-7- carboxylic acid (E3b).

5. A compound according to any one of claims 1 to 4 for use in medical therapy.

6. A pharmaceutical composition comprising a compound according to claim 1 to 4 and a pharmaceutically acceptable carrier.

7. A process for making a pharmaceutical composition comprising combining a compound according to any of the claims 1 to 4 and a pharmaceutically acceptable carrier.

8. A method of eliciting an nuclear hormone receptor modulating effect in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of a compound according to any of the claims l to 4.

9. The method according to claim 8 wherein the nuclear hormone receptor modulation effect is a receptor agonizing effect.

10. The method according to claim 8 wherein the nuclear hormone receptor modulation effect is a receptor antagonizing effect.

11. The method according to claim 8 wherein the nuclear hormone receptor modulation effect is a mixed receptor agonizing/antagonizing effect.

12. The method according to any of the claims 9 tol 1 wherein the nuclear hormone receptor modulation effect is an androgen receptor effect.

13. The method according to any of the claims 9 tol 1 wherein the nuclear hormone receptor modulation effect is an estrogen receptor effect.

14. The method according to any of the claims 9 tol 1 wherein the nuclear hormone receptor modulation effect is a glucocorticoid receptor effect.

15. The method according to any of the claims 9 tol 1 wherein the nuclear hormone receptor modulation effect is a mineralocorticoid receptor effect.

16. The method according to any of the claims 9 tol 1 wherein the nuclear hormone receptor modulation effect is a progesterone receptor effect.

17. The method according to any of the claims 9 tol 1 wherein the nuclear hormone receptor modulation effect is a liver X receptor effect.

18. The method according to any of the claims 9 tol 1 wherein the nuclear hormone receptor modulation effect is a peroxisome proliferator-activated receptor effect.

19. A method of treating or preventing a disease regulated by a nuclear receptor in a mammal in need thereof by administering to the mammal a therapeutically effective amount of a compound according to any of the claims 1 to 4.

20. A method of treating or preventing proliferative diseases, cancers, benign prostate hyperplasia, adenomas and neoplasisms of the prostate, benign or malignant tumor cells containing the androgen receptor, cardiovascular diseases, angiogenic conditions or disorders, hirsutism, acne, hyperpilosity, inflammation, immune modulation, seborrhea, endometriosis, polycycstic ovary syndrome, androgen alopecia, hypogonadism, osteoporosis, suppressing spermatogenesis, libido, cachexia, anorexia, cancers expressing the estrogen receptor, prostate cancer, breast cancer, endometrial cancer, incontinence, hot flashes, vaginal dryness, menopause, contraception, pregnancy termination, cancers containing the progesterone receptor, endometriosis, cachexia, menopause fibrosis, labor induction, autoimmune diseases, impairment of cognitive functioning, cerebral degenerative disorders, Alzheimer's disease, psychotic disorders, type-II diabetes, congestive heart failure, disregulation of cholesterol or lipid homeostasis, and/or obesity in a mammal in need thereof by administering to the mammal a therapeutically effective amount of a compound according to any of the claims 1 to 4.

21. The use of a compound according to any of the claims 1 to 4 in the manufacture of a medicament for the therapeutic treatment or prevention of proliferative diseases, cancers, benign prostate hyperplasia, adenomas and neoplasisms of the prostate, benign or malignant tumor cells containing the androgen receptor, cardiovascular diseases, angiogenic conditions or disorders, hirsutism, acne, hyperpilosity, inflammation, immune modulation, seborrhea, endometriosis, polycycstic ovary syndrome, androgen alopecia, hypogonadism, osteoporosis, suppressing spermatogenesis, libido, cachexia, anorexia, cancers expressing the estrogen receptor, prostate cancer, breast cancer, endometrial cancer, incontinence, hot flashes, vaginal dryness, menopause, contraception, pregnancy termination, cancers containing the progesterone receptor, endometriosis, cachexia, menopause fibrosis, labor induction, autoimmune diseases, impairment of cognitive functioning, cerebral degenerative disorders, Alzheimer's disease, psychotic disorders, type-II diabetes, congestive heart failure, disregulation of cholesterol or lipid homeostasis, and/or obesity.