WO2003074044A1 - Benzopyranes substitues utilises comme agonistes selectifs des recepteurs des oestrogenes beta - Google Patents

Benzopyranes substitues utilises comme agonistes selectifs des recepteurs des oestrogenes beta Download PDF

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WO2003074044A1
WO2003074044A1 PCT/US2003/002678 US0302678W WO03074044A1 WO 2003074044 A1 WO2003074044 A1 WO 2003074044A1 US 0302678 W US0302678 W US 0302678W WO 03074044 A1 WO03074044 A1 WO 03074044A1
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formula
compound
reaction
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PCT/US2003/002678
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Charles Willis Lugar, Iii
Jeffrey Alan Dodge
Venkatesh Gary Krishnan
Blake Lee Neubauer
Bryan Hurst Norman
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Eli Lilly And Company
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    • 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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2

Definitions

  • the present invention relates to novel cycloalkyl-benzopyrans and derivatives thereof, compositions containing those compounds, their use as selective estrogen receptor-beta agonists, and their use in the treatment of estrogen receptor-beta mediated diseases such as prostate cancer, benign prostatic hyperplasia, testicular cancer, ovarian cancer, lung cancer, cardiovascular diseases, neurodegenerative disorders, urinary incontinence, central nervous system (CNS) conditions, gastrointestinal (GI) tract conditions, and osteoporosis.
  • estrogen receptor-beta mediated diseases such as prostate cancer, benign prostatic hyperplasia, testicular cancer, ovarian cancer, lung cancer, cardiovascular diseases, neurodegenerative disorders, urinary incontinence, central nervous system (CNS) conditions, gastrointestinal (GI) tract conditions, and osteoporosis.
  • the estrogen receptor (ER) is currently the only member of the steroid subfamily of nuclear receptors that has different subtypes.
  • ER-beta also known as ER-betal
  • ER-alpha was cloned from a rat prostatic cDNA library and is present in murine and human prostates. Consequently, the previous ER is now designated as ER-alpha.
  • ER-alpha and ER-beta share high amino acid homology, have similar 17- ⁇ Estradiol (E2) binding affinities, and can hetero- or homodimerize to form a signaling complex; Kuiper GG, et al., Endocrinol. 138: 863-70 (1997); Kuiper GG et al., Proc. Natl. Acad. Sci. USA 93: 5925-30 (1996).
  • E2 activates both ER-alpha and ER-beta
  • ER-alpha stimulates transcription and cellular proliferation, while ER-beta suppresses ER-alpha activation.
  • 3-beta, 17-beta-androstanediol has been proposed to be an endogenous ligand for ER-beta; Weihua Z. et al. PNAS 98: 6330-5 (2001).
  • 3-Beta, 17-beta-androstanediol is a major metabolite of dihydrotestosterone (DHT), the 5-alpha- reduced active intracellular androgen in male accessory sex organs.
  • DHT dihydrotestosterone
  • ER-beta activation also stimulates increased glutathione S-transferase and quinone reductase expression.
  • ER-beta is strongly expressed in a number of tissues including prostate, bladder, ovary, testis, lung, small intestine, vascular endothelium, and various parts of the brain
  • compounds that selectively modulate ER-beta would be of clinical importance in the treatment of a variety of disease conditions, such as prostate cancer, testicular cancer, ovarian cancer, lung cancer, colon cancer, cardiovascular diseases, neurodegenerative disorders, urinary incontinence, CNS, anorexic behavior, GI tract conditions, and osteoporosis.
  • Such compounds would have minimal effect on tissues that contain ER-alpha, and thus exhibit different side-effect profiles.
  • ER- beta agonists will display different therapeutic profiles compared to ER-alpha antagonists or agonists, and would be preferentially beneficial in tissues relying on ER-beta signaling.
  • the prostate gland produces components that are found in the semen and blood. Some of these are regulatory peptides.
  • the prostate gland comprises stroma and epithelium cells, the latter group consisting of columnar secretory cells and basal non- secretory cells.
  • the proliferation of these basal cells, as well as stroma cells gives rise to benign prostatic hyperplasia (BPH), which is one common prostate disease.
  • BPH benign prostatic hyperplasia
  • Consequences of BPH can include hypertrophy of bladder smooth muscle, decompensated bladder, and increased incidence of urinary tract infection.
  • the development of BPH is considered to be an inescapable phenomenon for the aging male population. BPH is observed in approximately 70% of males over the age of 70.
  • Drug treatment for BPH currently employs alpha andrenergic antagonists for symptomatic relief or steroid 5 -alpha reductase inhibitors to reduce hyperplastic tissue bulk. These approaches are of limited therapeutic benefit.
  • R 1 , R 2 , R 3 , and R4 are each independently -H, C ⁇ -Cg alkyl, -OH, C ⁇ -Cg alkoxy, halo, or -CF 3 ; and ,
  • R5 and R 6 are each independently -H or C ⁇ -Cg alkyl; or a pharmaceutically salt thereof; and a pharmaceutically acceptable carrier.
  • Specific diseases mediated by estrogen receptor beta, and which are treated by compounds of formula I, include prostate cancer, benign prostatic hyperplasia, testicular cancer, ovarian cancer, lung cancer, cardiovascular diseases, neurodegenerative disorders, urinary incontinence, central nervous system (CNS) disorders, gastrointestinal (GI) tract disorders, and osteoporosis.
  • a preferred embodiment of the invention is said method wherein the compound of
  • Formula I is selected from the group consisting of
  • the present invention provides the novel substituted benzopyran compounds:
  • halogen refers to a fluorine atom, chlorine atom, bromine atom, or iodine atom
  • C ⁇ -Cg alkyl refers to a branched or straight chained alkyl radical containing from 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n- butyl, isobutyl, sec butyl, t-butyl, pentyl, hexyl, etc
  • C ⁇ -Cg alkoxy refers to a straight or branched alkoxy group containing from 1 to 6 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n- butoxy, isobutoxy, sec-butoxy, t-butoxy, pentoxy, hexoxy,
  • the compounds used in the method of the present invention may have one or more asymmetric centers.
  • the. compounds of the present invention occur as racemates and as individual enantiomers, as well as diastereomers and mixtures of diastereomers. All asymmetric forms, individual isomers and combinations thereof, are within the scope of the present invention.
  • the three main chiral centers signified as 1, 2, and 3, are illustrated in formula (I).
  • a number of routes are available. As an example, a mixture of enantiomers maybe prepared, and then the two enantiomers may be separated.
  • a commonly employed method for the resolution of the racemic mixture (or mixture of enantiomers) into the individual enantiomers is to first convert the enantiomers to diastereomers by way of forming a salt with an optically active acid or base. These diastereomers may then be separated using differential solubility, fractional crystallization, chromatography, or the like. Further details regarding resolution of enantiomeric mixtures maybe found in J. Jacques, et al., Enantiomers, Racemates, and Resolutions, (1991). "The term "pharmaceutically acceptable salts thereof refers to either an acid addition salt or a basic addition salt.
  • pharmaceutically acceptable acid addition salts is intended to apply to any non-toxic organic or inorganic acid addition salt of the base compounds represented by formula (I).
  • Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulphuric, and phosphoric acid and acid metal salts such as sodium monohydrogen orthophosphate, and potassium hydrogen sulfate.
  • Illustrative organic acids which form suitable salts include the mono-, di-, and tricarboxylic acids.
  • Such acids are for example, acetic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, benzoic, hydroxy-benzoic, phenylacetic, cinnamic, salicyclic, 2-phenoxy-benzo.ic, p- toluenesulfonic acid, and sulfonic acids such as benzenesulfonic acid, methanesulfonic acid, and 2-hydroxyethanesulfonic acid.
  • Such salts can exist in either a hydrated or substantially anhydrous form.
  • the acid addition salts of these compounds are soluble in water and various hydrophilic organic solvents, and which in comparison to their free base forms, generally demonstrate higher melting points.
  • compositions represented by formula (I) are intended to apply to any non-toxic organic or inorganic basic addition salts of the compounds represented by formula (I).
  • Illustrative bases which form suitable salts include alkali metal or alkaline-earth metal hydroxides such as sodium, potassium, calcium, magnesium, or barium hydroxides; ammonia, and aliphatic, alicyclic, or aromatic organic amines such as methylamine, dimethylamine, trimethylamine, and picoline. Either the mono- or di- basic salts can be formed with those compounds.
  • R la , R 2a , R 3a , and R 4a correspond to the substituents R 1 , R 2 , R 3 , and R 4 , respectively, except for when the R*, R 2 , R 3 , and R 4 substituents would be hydroxy.
  • the corresponding hydroxy group is protected with an alkoxymethylether, such as methoxymethyl ("MOM”) or methoxyethoxymethyl (“MEM").
  • step 1 the hydroxy groups on the phenol of formula (2) are protected with a suitable protecting group to provide the protected 2-bromo-phenol of formula (3) utilizing techniques and procedures well know to one of ordinary skill in the art.
  • the phenol of formula (2) is combined with a suspension comprising a suitable anhydrous solvent such as anhydrous dimethylform-amide (DMF) and a suitable strong base such as a metal hydride, most preferably sodium hydride.
  • a suitable anhydrous solvent such as anhydrous dimethylform-amide (DMF) and a suitable strong base such as a metal hydride, most preferably sodium hydride.
  • MOM alkoxymethyl ether
  • the reaction may be conducted at room temperature for a time ranging from about 30 minutes to about 2 days.
  • the reaction is then quenched with water and an appropriate ether, such as diethyl ether, and the organic layer is washed with an appropriate base, such as sodium hydroxide, and brine.
  • step 2 the protected phenol of formula (3) is coupled with the compound of formula (4) to provide the coupled product of formula (5).
  • the coupling reaction is conducted in the presence of butyllithium, zinc chloride, and a Pd species.
  • the reaction is preferably carried out in a suitable solvent such as tetrahydrofuran (THF), and may initially be carried out under anhydrous condition.
  • THF tetrahydrofuran
  • the protected phenol of formula (3) is dissolved in a suitable solvent such as THF, treated with butyllithium at reduced temperature, zinc chloride in solvent is then added and the temperature allowed to rise to ambient.
  • the palladium species such as tetrakis(friphenylphosphine)Pd(0)
  • the temperature is preferably raised to the reflux temperature of the solvent for a period of time from about 6 to 24 hours.
  • the coupled product of formula (5) can be isolated and'purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 3 the coupled product of formula (5) is reduced with a suitable reducing agent to provide the reduced product of formula (6) utilizing techniques and procedures well known in the art.
  • the coupled product of formula (5) is contacted with a suitable reducing agent, such as a palladium species, preferably 5% carbon on palladium, in a suitable solvent or solvent mixture, such as ethanol or methanol.
  • a suitable reducing agent such as a palladium species, preferably 5% carbon on palladium
  • a suitable solvent or solvent mixture such as ethanol or methanol.
  • the reaction is preferably carried out in the presence of a suitable base, such as a trialkylamine, more preferably, triethylamine.
  • the reaction mixture is then heated to a temperature ranging from about 30°C to about reflux for a period of time ranging from about 2 to 72 hours.
  • the reduced product of formula (6) can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 4 the reduced product of formula (6) can be converted to the Weinreb-amide of formula (7).
  • This reaction can be performed utilizing a reaction of the type described by J. M. Williams, et al., Tetrahedron Letters 36, 5461-5464 (1995).
  • the reduced product of formula (6) is combined with N,O- dimethylhydroxylamine hydrochloride in a suitable aprotic solvent, such as THF, preferably under anhydrous conditions and cooled to a temperature ranging from about 0°C to about -30°C, more preferably about -10°C.
  • a suitable Grignard reagent preferably isopropyl magnesium chloride, is then added in a molar ratio of about 1.5 and reaction mixture is stirred for about 1 hour to 100 hours. The reaction is then quenched with a proton source such as, for example, saturated potassium sodium tartrate aqueous.
  • a proton source such as, for example, saturated potassium sodium tartrate aqueous.
  • the Weinreb-amide of formula (7) can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization. In Scheme A, step 5, the Weinreb-amide of formula (7) is combined with the aryl lithium of formula (8) to form the ketone of formula (9).
  • the aryl lithium of formula (8) is added to a solution of Weinreb-amide of formula (7) in a suitable aprotic solvent, such as anhydrous THF, cooled to a temperature ranging from about -100°C to about 0°C, preferably -75°C, and stirred for a period of time ranging from about 1 hour to 30 hours.
  • a proton source such as, for example, saturated sodium potassium tartrate aqueous.
  • the ketone of formula (9) can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 6a the ketone of formula (9) is subjected to an acid-catalyzed cyclization followed by reduction of the resulting hemiketal to provide a compound of formula (10).
  • a suitable alcohol solvent such as anhydrous methanol.
  • the mixture is then heated at temperature ranging from 40°C to 60°C, preferably 50°C, for a period of time ranging from 12 to 24 hours, preferably 18 hours.
  • reaction is then cooled to ambient temperature and a suitable reducing agent, such as sodium cyanoborohydride, is added along with a suitable indicator such as bromocresol green in a procedure similar to that described by A. Srikrishna, et al., Tetrahedron, vol. 51, no. 11, pp. 3339-3344, 1995.
  • Methanol saturated with hydrochloric acid is then slowly added until a yellow color is maintained.
  • the reaction is stirred for about 1 to 2 hours past the point of final color change.
  • the reaction is then quenched with a suitable proton acceptor, such as saturated sodium potassium tartrate aqueous.
  • a suitable proton acceptor such as saturated sodium potassium tartrate aqueous.
  • step 6b The R3SiH/TFA conditions of step 6b will result predominantly in a trans-configuration of the chiral centers.
  • the product of formula (10) or (11) can then be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 1 a compound of formula (12) and a compound of formula (13) undergo a cyclization reaction to form a compound of formula (14) in the presence of an acid, preferably sulfuric acid.
  • step 2 the product of formula (14) is reduced with a suitable reducing agent to provide the reduced product of formula (15) utilizing techniques and procedures well known in the art.
  • a suitable reducing agent such as a palladium species, preferably 5% carbon on palladium
  • a suitable solvent or solvent mixture such as ethanol or methanol.
  • the reaction is preferably carried out in the presence of a suitable base, such as a frialkylamine, more preferably, triethylamine.
  • the reaction mixture is then heated to a temperature ranging from about 30°C to about reflux for a period of time ranging from about 2 to 72 hours.
  • the reduced product of formula (14) can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 3 the reduced product of formula (15) is combined with an aryl lithium to form the compound of formula (17).
  • the aryl lithium of formula (16) is added to a solution formula (15) in a suitable aprotic solvent, such as anhydrous THF, cooled to a temperature ranging from about -100°C to about 0°C, preferably -75°C, and stirred for a period of time ranging from about 1 hour to 30 hours.
  • the reaction is then quenched with a proton source, such as, for example, saturated sodium potassium tartrate aqueous.
  • the compound of formula (17) can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 4a the compound of formula (17) is subjected to a reduction to provide a compound of formula (18).
  • p-toluenesulfonic acid is added in roughly equimolar proportions to the ketone of formula (17) in a suitable alcohol solvent, such as anhydrous methanol.
  • a suitable alcohol solvent such as anhydrous methanol.
  • the mixture is then heated at temperature ranging from 40°C to 60°C, preferably 50°C, for a period of time ranging from 12 to 24 hours, preferably 18 hours.
  • the reaction is then cooled to ambient temperature and a suitable reducing agent, such as sodium cyanoborohydride, is added along with a suitable indicator such as bromocresol green in a procedure similar to that described by A. Srikrishna, et.
  • step 4a Methanol saturated with hydrochloric acid is then slowly added until a yellow color is maintained. The reaction is stirred for about 1 to 2 hours past the point of final color change. The reaction is then quenched with a suitable proton acceptor, such as saturated sodium potassium tartrate aqueous.
  • a suitable proton acceptor such as saturated sodium potassium tartrate aqueous.
  • This set of reaction conditions for step 4a will result in a cis-configuration of the chiral centers.
  • the R3SiH/TFA conditions of step 4b will result in a trans- configuration of the chiral centers.
  • the product of formula (18) or (19) can then be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step la the hydroxy groups on the phenol of formula (20) are protected with a suitable protecting group to provide the protected phenol of formula (21) utilizing techniques and procedures well know to one of ordinary skill in the art.
  • step lb a compound of formula (22) is converted to the triflate to provide the compound of formula (23) utilizing procedures and techniques well known in the art; G. T. Crisp et al., J. Ore. Chem. 57, 6972-6975 (1992).
  • the carboxylate of formula (22) is dissolved under anhydrous conditions in a suitable solvent, such as tetrahydrofuran, dichloromethane, acetone, ethyl acetate, toluene, or diethyl ether and contacted with a suitable activating agent such as triflic anhydride.
  • a suitable solvent such as tetrahydrofuran, dichloromethane, acetone, ethyl acetate, toluene, or diethyl ether
  • a suitable activating agent such as triflic anhydride.
  • the reaction is carried out in the presence of a base, such as N-methylmorpholine, sodium carbonate, triethylamine, N,N-diisopropylethylamine, potassium carbonate or sodium bicarbonate.
  • the reaction is generally carried out at temperatures of from -78°C to ambient temperature. Generally, the reactions require 1 to 24 hours. The reaction may then be quenched.
  • step 2 the protected phenol of formula (21) is coupled with the activated carboxylate of formula (23) to provide the coupled product of formula (24).
  • the coupling reaction is conducted in the presence of butyllithium, zinc chloride and a Pd species.
  • the reaction is preferably carried out in a suitable solvent such as tetrahydrofuran (THF), and may initially be carried out under anhydrous condition.
  • THF tetrahydrofuran
  • the protected phenol of formula (21) is dissolved in a suitable solvent such as THF, treated with butyllithium at reduced temperature, zinc chloride in solvent is then added and the temperature allowed to rise to ambient.
  • the palladium species such as tetrakis(triphenylphosphine)Pd(0)
  • the temperature is preferably raised to the reflux temperature of the solvent for a period of time ranging from about 6 to 24 hours.
  • the coupled product of formula (24) can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 3 the coupled product of formula (24) is reduced to the product of formula (25). The reduction can be performed by the method described for Scheme A, step 3.
  • step 4 the product of formula (25) is can be converted to the Weinreb-amide of formula (26).
  • This reaction can be performed utilizing a reactin of the type described by J. M. Williams, et al., Tetrahedron Letters 36, 5461-5464 (1995). The reaction can be performed by the method described for Scheme A, step 4.
  • step 5 the Weinreb-amide of formula (26) is combined with the aryl lithium of formula (27) to form the ketone of formula (28).
  • the reaction can be performed by the method described for Scheme A, step 5.
  • step 6a the ketone of formula (28) is subjected to an acid-catalyzed cyclization followed by reduction of the resulting hemiketal to provide a compound of formula (29).
  • the R3SiH/TFA conditions of step 6b will result in a trans-configuration of the chiral centers forming a compound of the formula (30).
  • Steps 6a and 6b can be performed by the method as described for Scheme A, step 6a and 6b, respectively.
  • step 3 p-toluenesulfonic acid tetrabutylammonium cyanoborohydride MeOH saturated with HCl gas
  • step 1 the tetrahydrocoumarin compound of formula (31) is protected with triisopropylsilyl chloride to form the protected compound of formula (32).
  • the reaction is performed in a suitable solvent such as methylene chloride and diisopropylethylamine.
  • the reaction is generally performed at ambient temperature from 70 to 150 hours, but typically at 125 hours.
  • the reaction is then quenched typically with saturated sodium bicarbonate aqueous.
  • the product of formula (32) can then be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 2 the protected compound of formula (32) is combined with the compound of formula (33) to form the compound of formula (34).
  • the product of formula (34) can then be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • step 3 the compound of formula (34) is deprotected and reduced to form a compound of formula (35).
  • the product of formula (35) can then be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, chromatography, and recrystallization.
  • Example 1 was dried over anhydrous sodium sulfate, concentrated in vacuo, and slurried with 2% methanol/methylene chloride to give 0.03 g (29%) of the Example 1 as a -6:1 cis:trans stereoisomer mixture.
  • Example 4 may be made by the methods of Preparations 9, 10, 11, 12 and Example 3 by substituting n-Butyl-2-ethylacrylate for n- Butyl-2-methylacrylate as a starting material in Preparation 9.
  • the competition ER binding assay was run in a buffer containing 50 mM N-[2- hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid (Hepes) pH 7.5, 1.5 mM EDTA, 150 mM NaCl, 10% glycerol, 1 mg/mL ovalbumin, 5mM DTT, 0.025 ⁇ Ci per well of 3 H- Estradiol(NEN #NET517 at 118 Ci/mmol, 1 mCi/mL), and 10 ng/well ERAlpha or ERbeta Receptor (PanNera). Competing compounds were added at 10 different concentrations.
  • Non-specific binding was determined in the presence of 1 ⁇ M of E2 (17- ⁇ Estradiol, Sigma, St. Louis, MO).
  • the binding reaction 140 ⁇ L was incubated for 4 hours at room temperature, then 70 ⁇ L of cold dextran coated charcoal (DCC) buffer was added to each reaction (DCC buffer was prepared by adding 0.75g of charcoal [Sigma] and 0.25g of dextran [Pharmacia] per 50 mL of assay buffer).
  • DCC buffer was prepared by adding 0.75g of charcoal [Sigma] and 0.25g of dextran [Pharmacia] per 50 mL of assay buffer).
  • the incubation plates were mixed for 8 minutes on an orbital shaker at 4°C and then centrifuged at 3,000 rpm for 10 minutes at 4°C.
  • Compounds of Examples 1-4 are active in the assay as described.
  • the compounds of Examples 1-4 bind to the ER beta receptor with a Kj of less than 100 nM.
  • Preferred compounds bind to the ER beta receptor with a Kj of less than 30 nM. More preferred compounds bind to the ER beta receptor with a Kj of less than 2 nM.
  • Compounds that are selective to binding to the ER beta receptor compared to the ER alpha receptor bind to the ER beta receptor with a lower Kj compared to the K for the ER alpha receptor.
  • Preferred selective ER beta compounds bind to ER beta receptor with a K;(ER alpha)/Kj(ER beta) ratio of greater than 4 as shown in Table 1.
  • ERbeta agonists are evaluated for their effects on the growth of androgen-sensitive LNCaP human prostatic cancer (PCa) xenografts grown in intact sexually mature (5-6 weeks old) Hsd: Athymic Nude-nu (Athymic Nude) male mice. 2.0x10 6 LNCaP tumor cells are injected bilaterally by the subcutaneous route into the pre-tracheal region of testicular intact male mice. Mice are castrated via the scrotal route to serve as the positive control group. Test compounds are administered once per day by subcutaneous or gavage administration at multiple dose levels in a volume of 0.2 ml to xenograft-bearing mice starting on the day following tumor injection.
  • PCa human prostatic cancer
  • Test compounds are reformulated weekly based on average group mean body weights.
  • the vehicle for these studies is 1%) carboxymethyl cellulose (CMC) with 0.25% Tween 80.
  • CMC carboxymethyl cellulose
  • Tween 80 0.25% Tween 80.
  • Body weights and tumor measurements are recorded on a weekly basis and entered directly into a JMPTM (SAS; Gary, NC) spreadsheet from electronic caliper measurement.
  • Tumor volumes in mm 3 are calculated in JMP using the following formula: L X W X H X 0.5236.
  • Tumor and body weight responses for individual mice are recorded on a weekly basis. When LNCaP tumor volumes enter log-phase expansion, lesions are measured every 3-4 days.
  • SAS linear extrapolation model
  • mice Seventy-five day old (unless otherwise indicated) female Sprague Dawley rats (weight range of 200 to 225 g) are obtained from Charles River Laboratories (Portage, MI). The animals are either bilaterally ovariectomized (OVX) or exposed to a Sham surgical procedure at Charles River Laboratories, and then shipped after one week. Upon arrival, they are housed in metal hanging cages in groups of 3 or 4 per cage and have ad libitum access to food (calcium content approximately 0.5%) and water for one week. Room temperature is maintained at 22.2° ⁇ 1.7°C with a minimum relative humidity of 40%. The photoperiod in the room was 12 hours light and 12 hours dark. Dosing Regimen Tissue Collection: After a one week acclimation period
  • Serum cholesterol is determined using a Boehringer Mannheim Diagnostics high performance cholesterol assay. Briefly the cholesterol is oxidized to cholest-4-en-3-one and hydrogen peroxide. The hydrogen peroxide is then reacted with phenol and 4-aminophenazone in the presence of peroxidase to produce a p-quinone imine dye, which is read spectrophotemetrically at 500 nm. Cholesterol concentration is then calculated against a standard curve. The entire assay is automated using a Biomek Automated Workstation.
  • the uteri from above are kept at 4°C until time of enzymatic analysis.
  • the uteri are then homogenized in 50 volumes of 50 mM Tris buffer (pH 8.0) containing 0.005% Triton X- 100.
  • Tris buffer pH 8.0
  • Upon addition of 0.01 % hydrogen peroxide and 10 mM O- phenylenediamine (final concentrations) in Tris buffer increase in absorbance is monitored for one minute at 450 nm.
  • the presence of eosonophils in the uterus is an indication of estrogenic activity of a compound.
  • the maximal velocity of a 15 second interval is determined over the initial, linear portion of the reaction curve.
  • the rats are treated daily for thirty-five days (6 rats per treatment group) and sacrificed by carbon dioxide asphyxiation on the 36th day.
  • the thirty-five day time period is sufficient to allow maximal reduction in bone density, measured as described herein.
  • the uteri are removed, dissected free of extraneous tissue, and the fluid contents are expelled before determination of wet weight in order to confirm estrogen deficiency associated with complete ovariectomy. Uterine weight is routinely reduced about 75%) in response to ovariectomy.
  • the uteri are then placed in 10% neutral buffered formalin to allow for subsequent histological analysis.
  • the right femurs are excised and digitilized X-rays generated and analyzed by an image analysis program (NIH image) at the distal metaphysis.
  • NASH image image analysis program
  • the proximal aspect of the tibiae from these animals are also scanned by quantitative computed tomography.
  • F-I or ethynyl estradiol (EE2) in 20%» hydroxypropyl ⁇ -cyclodextrin are orally administered to test animals.
  • Narious diseases and conditions described to be treated herein, are well known and appreciated by those skilled in the art. It is also recognized that one skilled in the art may affect the associated diseases and conditions by treating a patient presently afflicted with the diseases or conditions or by prophylactically treating a patient afflicted with the diseases or conditions with a therapeutically effective amount of the compounds of - formula (I).
  • the term "patient” refers to a warm blooded animal such as a mammal which is afflicted with a particular inflammatory disease state. It is understood that guinea pigs, dogs, cats, rats, mice, horses, cattle, sheep, and humans are examples of animals within the scope of the meaning of the term.
  • the term "therapeutically effective amount" of a compound of formula (I) refers to an amount which is effective in controlling diseases and conditions associated with estrogen receptor-beta mediated diseases such as prostate cancer, benign prostatic hyperplasia, testicular cancer, ovarian cancer, lung cancer, cardiovascular diseases, neurodegenerative disorders, urinary incontinence, central nervous system (C ⁇ S) conditions, gastrointestinal (GI) tract conditions, and osteoporosis.
  • estrogen receptor-beta mediated diseases such as prostate cancer, benign prostatic hyperplasia, testicular cancer, ovarian cancer, lung cancer, cardiovascular diseases, neurodegenerative disorders, urinary incontinence, central nervous system (C ⁇ S) conditions, gastrointestinal (GI) tract conditions, and osteoporosis.
  • controlling is intended to refer to all processes wherein there may be a slowing, interrupting, arresting, or stopping of the progression of the diseases and conditions described herein, but does not necessarily indicate a total elimination of all disease and condition symptoms, but does include prophylactic treatment of the diseases and conditions associated with estrogen receptor-beta mediated diseases such as prostate cancer, benign prostatic hyperplasia, testicular cancer, ovarian cancer, lung cancer, cardiovascular diseases, neurodegenerative disorders, urinary incontinence, central nervous system (C ⁇ S) conditions, gastrointestinal (GI) tract conditions, and osteoporosis.
  • a therapeutically effective amount can be readily determined by the attending diagnostician, as one skilled in the art, by the use of conventional techniques and by observing results obtained under analogous circumstances.
  • the dose a number of factors are considered by the attending diagnostician, including, but not limited to: the species of mammal; its size, age, and general health; the specific disease involved; the degree of or involvement or the severity of the disease; the response of the individual patient; the particular compound administered; the mode of administration; the bioavailability characteristic of the preparation administered; the dose regimen selected; the use of concomitant medication; and other relevant circumstances.
  • a therapeutically effective amount of a compound of formula (I) is expected to vary from about 0.001 milligram per kilogram of body weight per day (mg/kg/day) to about 100 mg/kg/day. Preferred amounts can be determined by one skilled in the art.
  • a compound of formula (I) can be administered in any form or mode which makes the compound bioavailable in a therapeutically effective amount, including oral, inhalation, and parenteral routes.
  • compounds of formula (I) can be administered orally, by inhalation of an aerosol or dry powder, subcutaneously, intramuscularly, intravenously, fransdermally, intranasally, rectally, topically, and the like.
  • Oral or inhalation administration is generally preferred for treatment of respiratory diseases, e.g. asthma.
  • respiratory diseases e.g. asthma.
  • One skilled in the art of preparing formulations can readily select the proper form and mode of administration depending upon the particular characteristics of the compound selected, the disease or condition state to be treated, the stage of the disease or condition, and other relevant circumstances. (Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Co. (1990)).
  • the compounds of the present invention can be administered alone or in the form of a pharmaceutical composition in combination with pharmaceutically acceptable carriers or excipients, the proportion and nature of which are determined by the solubility and chemical properties of the compound selected, the chosen route of administration, and standard pharmaceutical practice.
  • the compounds of the present invention while effective themselves, may be formulated and administered in the form of their pharmaceutically acceptable salts, such as acid addition salts or base addition salts, for purposes of stability, convenience of crystallization, increased solubility and the like.
  • the present invention provides pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula (I) in admixture or otherwise in association with one or more pharmaceutically acceptable carriers or excipients.
  • the pharmaceutical compositions are prepared in a manner well known in the pharmaceutical art.
  • the carrier or excipient may be a solid, semi-solid, or liquid material which can serve as a vehicle or medium for the active ingredient. Suitable carriers or excipients are well known in the art.
  • the pharmaceutical composition may be adapted for oral, inhalation, parenteral, or topical use and may be administered to the patient in the form of tablets, capsules, aerosols, inhalants, suppositories, solution, suspensions, or the like.
  • the compounds of the present invention may be administered orally, for example, with an inert diluent or with an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets.
  • the compounds may be incorporated with excipients and used in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, chewing gums and the like.
  • These preparations should contain at least 4% of the compound of the present invention, the active ingredient, but may be varied depending upon the particular form and may conveniently be between 4% to about 70% of the weight of the unit.
  • the amount of the compound present in compositions is such that a suitable dosage will be obtained.
  • Preferred compositions and preparations according to the present invention may be ⁇ determined by someone skilled in the art.
  • the tablets, pills, capsules, troches and the like may also contain one or more of the following adjuvants: binders such as microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch or lactose, disintegrating agents such as alginic acid, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; and sweetening agents such as sucrose or saccharin may be added or a flavoring agent such as peppermint, methyl salicylate or orange flavoring.
  • a liquid carrier such as polyethylene glycol or a fatty oil.
  • dosage unit forms may contain other various materials which modify the physical form of the dosage unit, for example, as coatings.
  • tablets or pills may be coated with sugar, shellac, or other enteric coating agents.
  • a syrup may contain, in addition to the present compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors. Materials used in preparing these various compositions should be pharmaceutically pure and non-toxic in the amounts used.
  • the compounds of the present invention may be incorporated into a solution or suspension. These preparations should contain at least 0.1 % of a compound of the invention, but may be varied to be between 0.1 and about 50% of the weight thereof. The amount of the compound of formula (I) present in such compositions is such that a suitable dosage will be obtained. Preferred compositions and preparations are able to be determined by one skilled in the art.
  • the compounds of the present invention may also be administered by inhalation, such as by aerosol or dry powder. Delivery may be by a liquefied or compressed gas or by a suitable pump system which dispenses the compounds of the present invention or a formulation thereof.
  • Formulations for administration by inhalation of compounds of formula (I) may be delivered in single phase, bi-phasic, or tri-phasic systems.
  • a variety of systems are available for the administration by aerosols of the compounds of formula (I).
  • Dry powder formulations are prepared by either pelletizing or milling the compound of formula (I) to a suitable particle size or by admixing the pelletized or milled compound of formula (I) with a suitable carrier material, such as lactose and the like.
  • Delivery by inhalation includes the necessary container, activators, valves, subcontainers, and the like.
  • Preferred aerosols and dry powder formulations for administration by inhalation are able to be determined by one skilled in the art.
  • the compounds of the present invention may also be administered topically, and when done so the carrier may suitably comprise a solution, ointment or gel base.
  • the base for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers.
  • Topical formulations may contain a concentration of the formula (I) or its pharmaceutical salt from about 0.1 to about 10%o w/v (weight per unit volume).
  • the solutions or suspensions may also include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylene diaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose.
  • the parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

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  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
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Abstract

La présente invention concerne des dérivés de benzopyranes substitués, leurs stéréo-isomères, et leurs sels pharmaceutiquement acceptables. Les composés de l'invention sont utiles comme agonistes des récepteurs des oestrogènes bêta. Ces agonistes peuvent être utilisés pour le traitement de maladies induites par les récepteurs des oestrogènes bêta, telles que le cancer de la prostate.
PCT/US2003/002678 2002-03-01 2003-02-13 Benzopyranes substitues utilises comme agonistes selectifs des recepteurs des oestrogenes beta WO2003074044A1 (fr)

Priority Applications (1)

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AU2003212856A AU2003212856A1 (en) 2002-03-01 2003-02-13 Substituted benzopyrans as selective estrogen receptor-beta agonists

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US36152402P 2002-03-01 2002-03-01
US60/361,524 2002-03-01

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US9078888B2 (en) 2007-01-22 2015-07-14 Gtx, Inc. Nuclear receptor binding agents
US9604931B2 (en) 2007-01-22 2017-03-28 Gtx, Inc. Nuclear receptor binding agents
US9623021B2 (en) 2007-01-22 2017-04-18 Gtx, Inc. Nuclear receptor binding agents

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WO2000062765A2 (fr) * 1999-04-16 2000-10-26 Astrazeneca Ab β-LIGANDS DE RECEPTEURS D'OESTROGENE
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WO2000062765A2 (fr) * 1999-04-16 2000-10-26 Astrazeneca Ab β-LIGANDS DE RECEPTEURS D'OESTROGENE
WO2001064665A1 (fr) * 2000-03-01 2001-09-07 Akzo Nobel N.V. Derives de chromane en tant que composes oestrogeniques

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9078888B2 (en) 2007-01-22 2015-07-14 Gtx, Inc. Nuclear receptor binding agents
US9604931B2 (en) 2007-01-22 2017-03-28 Gtx, Inc. Nuclear receptor binding agents
US9623021B2 (en) 2007-01-22 2017-04-18 Gtx, Inc. Nuclear receptor binding agents

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