Patent Application: US-29731002-A

Abstract:
the present invention comprises the design , synthesis and development of a new class of chemotherapeutic agents for prophylactic and therapeutic treatments in a mammal , particularly a human , believed to be at risk of suffering from a hormone - responsive disorde . in an embodiment of the invention , such treatments include therapeutic compositions comprising novel steroidal antiestrogen and antiandrogen compounds . in a preferred embodiment , such a novel compound of the present invention has an address and a message component , which are made into a single composite entity for more aggressive intervention and effective treatment of hormone - responsive disorders , thereby prolonging the disease - free interval for the patient and reducing a number of side effects .

Description:
the term “ alkyl ” used herein refers to a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms , such as methyl , ethyl , n - propyl , isopropyl , n - butyl , isobutyl , t - butyl , octyl , decyl , tetradecyl , hexadecyl , eicosyl , tetracosyl and the like . preferred alkyl groups herein contain 1 to 12 carbon atoms . the term “ lower alkyl ” intends an alkyl group of one to six carbon atoms , preferably one to four carbon atoms . the term “ cycloalkyl ” intends a cyclic alkyl group , typically of 3 to 6 carbon atoms , more preferably 4 to 5 carbon atoms . the term “ cyclooxyalkyl ” intends a cyclic alkyl group containing a single ether linkage , again , typically containing 3 to 6 carbon atoms , more preferably 4 to 5 carbon atoms . the term “ aryl ” as used herein refers to a monocyclic aromatic species of 5 to 7 carbon atoms , and is typically phenyl . optionally , these groups are substituted with one to five , more preferably one to three , lower alkyl , lower flouroalkyl , lower alkoxy , halo , nitro , amino , amide , carboxy , thioether , sulfide , sulfoxide , sulfamino , and / or sulfamide substituents . the aryl group may also comprise of di -, tri -, hexa -, penta - substituted phenyl with all positional ( ortho , meta , para ) variations . the term “ lower flouroalkyl ” intends an alkyl group of one to six carbon atoms , preferably one to four carbon atoms . the term “ lower alkoxy ” intends an alkoxy group with one to six carbon atoms , preferably one to four carbon atoms . the term “ carboxy aryl ” as used herein refers to a carboxy group attached to the aryl group . the term “ halo ” or “ halogen ” refers to fluoro , chloro , bromo or iodo , and usually relates to halo substitution for a hydrogen atom in an organic compound . the term “ optional ” or “ optionally ” means that the subsequently described event or circumstance may or may not occur , and that the description includes instances where said event or circumstance occurs and instances where it does not . the term “ heteroaryl ” as used herein refers to monocyclic aromatic species of three to seven carbon atoms , and is preferably one to six carbon atoms , and is more preferably one to five carbon atoms , and is typically phenyl . in particular , the heteroaryl comprises , for example , oxazole , thiazole , isoxazole , where these heteroaryls have nitrogen , oxygen , or sulfur atoms in the monocyclic ring . optionally , these groups are substituted with one to five , more preferably one to three , lower alkyl , lower flouroalkyl , lower alkoxy , halo , nitro , amino , amide , carboxy , thioether , sulfide , sulfoxide , sulfamino , and / or sulfamide substituents . the aryl group may also comprise of di -, tri -, hexa -, penta - substituted phenyl with all positional ( ortho , meta , para ) variations . the term “ carboxy - heteroaryl ” as used herein refers to a carboxy group attached to a heteroaryl group . the term “ fused aryl ” as used herein refers to bicyclic aromatic species of three to seven carbon atoms , and is typically phenyl . in particular , the fused aryl may comprise of naphthyl , benzothienyl , or benzofuryl . optionally , these groups are substituted with one to five , more preferably one to three , lower alkyl , lower flouroalkyl , lower alkoxy , halo , nitro , amino , amide , carboxy , thioether , sulfide , sulfoxide , sulfamino , and / or sulfamide substituents . the aryl group may also comprise of di -, tri -, hexa -, penta - substituted phenyl with all positional ( ortho , meta , para ) variations . the term “ carboxy - fused aryl ” as used herein refers to a carboxy group attached to a fused - aryl group . the term “ biaryl ” as used herein refers to two monocyclic aromatic species of four to seven carbon atoms , and is typically different configurations of a combination of a phenyl and a heteroaryl . optionally , these groups are substituted with one to five , more preferably one to three , lower alkyl , lower flouroalkyl , lower alkoxy , halo , nitro , amino , amide , carboxy , thioether , sulfide , sulfoxide , sulfamino , and / or sulfamide substituents . the aryl group may also comprise of di -, tri -, hexa -, penta - substituted phenyl with all positional ( ortho , meta , para ) variations . the term “ carboxy - biaryl ” as used herein refers to a biaryl attached to a carboxy group . the terms “ ether - linked aryls ” and “ ether - linked heteroaryls ” as used herein refer to two aryls / heteroaryls as defined above that are linked by an ether group . optionally , these groups are substituted with one to five , more preferably one to three , lower alkyl , lower flouroalkyl , lower alkoxy , halo , nitro , amino , amide , carboxy , thioether , sulfide , sulfoxide , sulfamino , and / or sulfamide substituents . the aryl group may also comprise of di -, tri -, hexa -, penta - substituted phenyl with all positional ( ortho , meta , para ) variations . the terms “ amine - linked aryls ” and “ amine - linked heteroaryls ” as used herein refer to two aryls / heteroaryls as defined above that are linked by an amine group . the terms aminoalkoxyl arene hybrids and peptidyl hybrids as used herein are referred to the groups exemplified in table 1 . optionally , these groups are substituted with one to five , more preferably one to three , lower alkyl , lower flouroalkyl , lower alkoxy , halo , nitro , amino , amide , carboxy , thioether , sulfide , sulfoxide , sulfamino , and / or sulfamide substituents . aryl group may also comprise of di -, tri -, hexa -, penta - substituted phenyl with all positional ( ortho , meta , para ) variations . the term “ effective amount ” as used herein means a nontoxic but sufficient amount of a compound to provide the desired effect . the exact amount required will vary from patient to patient , depending on the species , age , and general condition of the patient , the severity of the condition being treated , and the particular compound and its mode of administration . thus , it is not possible to specify an exact “ effective amount .” however , an appropriate effective amount may be determined by one of ordinary skill in the art using only routine experimentation . the term “ pharmaceutically acceptable ” as used herein means a material which is not biologically or otherwise undesirable , i . e ., the material may be administered to a patient along with the selected compound without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained . the present invention comprises the design , synthesis and development of a new class of chemotherapeutic agents for the treatment of hormone - responsive disorders . in the new class of chemotherapeutic agents , two components — a message subunit or pharmacophore , present in the nonsteroidal antagonists ( e . g ., antiandrogens , antiestrogens ) and the address subunit found in the steroidal agonists ( e . g ., androgens , estrogens )— are combined into a single composite entity . in particular , specific compounds in this new class of chemotherapeutic agents target the estrogen and / or the androgen receptors . the general formula for the agents of the invention was determined based on the discovery that the interaction between androgen / estrogen with the receptor involves a two step process . there is an initial association of the hormone ( address component ) with a specific part of the receptor , called the hormone binding domain , followed by the induction of a conformational change in the receptor ( message component ) that generates the observed biological response . accordingly , the present invention incorporates the “ address - message ” concept to generate , for example , prostate cancer tissue affinity , selectivity , and efficacy , and employs transition metal catalysts / reagents to prepare the novel therapeutic compounds . as an embodiment , the present invention uses modified palladium catalysts for carbon - carbon ( stille , suzuki reactions ) and carbon - nitrogen / oxygen ( buchwald , hartwig ) coupling reactions . as another embodiment of the present invention , the use of 1d / 2d - nmr ( nuclear magnetic resonance ) and the molecular modeling in the evaluation of the conformational analysis of the target compounds provides the capability for biological and structural data . the novel therapeutic compounds constitute a structurally unique class of steroidal derivatives , e . g ., derivatives of , for example , 17α -( substituted ) phenylvinyl - 17β - estradiols as estrogens and antiestrogens , and corresponding ( nor ) testosterones and dihydro - derivatives . in particular , identification of the most potent and selective antagonists for prophylaxis and treatment provides for a more effective treatment of hormone - responsive disorders and thereby prolong the disease - free interval . the present invention provides for a more potent and effective agent which increases the initial response coupled with a slower progression to hormone independence . additionally , the therapeutic compound of the present invention targets specifically and more selectively thereby reducing the incidence and / or severity of the side effects of anti - estrogen or anti - androgen therapy . the following examples are presented to illustrate the advantages of the present invention and to assist one of ordinary skill in making and using the same . these examples are not intended in any way otherwise to limit the scope of the disclosure . preferred antiestrogens / antiandrogens for the prevention or treatment of its corresponding hormone - related disorder acting to inhibit estrogen / androgen action may be prepared accordingly as follows : example i synthesis and evaluation of steroidal antiestrogens at the 17α - position of estradiol reagents and solvents were obtained from commercial sources ( aldrich and sigma ) and were used without further purification . wang resins and carboxylated polystyrene resins were obtained from novabiochem . the loading capacities of the resins , 0 . 75 mmol g − 1 for the wang resin and 2 . 47 mmol g − 1 for the polystyrene resin , were determined by the manufacturer . a specially designed flask which had a glass frit , through which the reaction mixture could be filtered by applying pressure , was used for the solid phase synthesis . purifications for the intermediates were done by rinsing resins three times with the following solvents : ch 2 cl 2 , thf , dmf , meoh , ch 2 cl 2 . the cleaved products were purified on a silica gel column chromatography using the appropriate solvents and were characterized by melting point , nmr , ir and electrical analysis . melting points were determined in open capillary on an electrothermal melting point apparatus and were uncorrected . ir spectra were recorded on a perkin - elmer model 1600 ft - ir spectrometer . 1 h and 13 c nmr spectra were obtained with a varian xl - 300 nmr spectrometer at 300 mhz in cdcl 3 , acetone - d 6 , or dmso - d 6 as a solvent . elemental analyses were performed by atlantic microlab , inc . ( norcross , ga .). as on - resin reaction monitoring methods , color tests and ft - ir methods were used . bomoscresol green ( 0 . 5 % in ethanol , ph = 8 ) was used to assay for free carboxylic acids . 18 the color of the stock solution was dark blue and changed to yellow in the presence of free carboxy groups . antimony ( iii ) chloride solution ( 25 % in ccl 4 ) was also used to determine whether the steroid ( 17α - ethynyl estradiol ) was coupled to the resin and a positive test result for the presence of estradiol was indicated by the color purple ( carr , 1926 ; blatz , 1972 ; jork , 1990 ). in addition , a spectro - scopic method ( ft - ir ) was facilitated to detect chromophore change by reaction . ( method a ). the wang resins ( 1 g , 0 . 75 mmol ) were swelled in the ch 2 cl 2 overnight and rinsed twice with thf , ch 3 oh , ch 2 cl 2 and acetone . acetone ( 5 ml ) was added to the swelled resins . to the slurry was added 1 ml of jones reagent ( bowden , 1946 ) in a dropwise manner . the mixture was allowed to stand at room temperature for 24 h . the resin mixture was rinsed twice with water - acetone ( 1 : 1 ), ch 3 oh , dmf , dmso and ch 2 cl 2 and dried in vacuo . the loading capacity after the carboxylation reaction was 0 . 4 - 0 . 6 mmol g − 1 , which was determined with the coupling of 17α - ethynyl estradiol to the resin . the aliquot of the resins was characterized by ft - ir . ft - ir ( kbr ) v : 3000 - 3500 ( oh , broad ), 1690 ( c ═ o , broad ), 1603 , 1492 , 1452 ( aromatic ring ), 1279 ( c — o ). ( method b ). the carboxylation of a polystyrene resin was accomplished using the method described by farrall et al . ( farrall , 1976 ). ft - ir ( kbr ) v : 3420 ( oh , broad ), 1630 ( c ═ o , broad ), 1200 - 1400 ( c — o , broad ). loading capacity : 1 . 5 - 1 . 9 mmol g − 1 . the carboxylated wang resin ( 2 . 3 g ) or polystyrene resin ( 2 . 5 g ) was placed in the reactor equipped with a magnetic stirrer . the resin was swelled in the ch 2 cl 2 for 5 h and washed sequentially with thf , dmf , ch 3 oh , thf and ch 2 cl 2 . to the resin was added 0 . 23 g ( 1 . 1 mmol ) of dicyclohexylcarbodiimide ( ddc ) and 5 ml of ch 2 cl 2 and the mixture was mildly stirred for 10 min . to the slurry was added 0 . 75 g ( 2 . 6 mmol ) of 17α - ethynyl estradiol dissolved in 10 ml of ch 2 cl 2 - dmf ( 9 : 1 ) solvent and catalytic amount of 4 - dimethylaminopyridine ( dmap ). the reaction mixture was stirred for 5 min and then allowed to stand at room temperature for 24 h . the resin was washed three times with ch 2 cl 2 ch 3 oh , ipa ( 60 ° c . ), thf and dmf ( 60 ° c .) ( morales , 1998 ). the rinsed resin was dried under vacuum for 5 h . the actual loading of the resin was determined by quantitative measurement of the material by cleavage from known weight of resin using 5 n - naoh in ch 3 oh - dioxane ( 1 : 3 ). the resin - bound steroids were characterized by ft - ir and the cleaved compounds by h and [ 0102 ] 13 c nmr before proceeding to the next step . the loading capacity of each resin was shown in method a and b ; ft - ir ( kbr ) v ; 3437 ( 17β — oh ), 3301 ( 17α - c ≡ c - h ), 1735 ( c ═ o ), 1607 , 1493 , 1452 ( aromatic ring ), 1216 ( c — o ). ( method a ). the 17α - ethynyl estradiol coupled to the resin ( 0 . 49 g , 0 . 57 mmol g − 1 ) was placed in a dry 25 ml reaction flask equipped with a reflux condenser and a magnetic stirrer and was swelled in thf for 1 h . to the slurry in the dry thf were treated triethylborane ( 0 . 7 ml ) and tributyltin hydride ( 1 ml ) ( nozaki , 1989 ). the mixture was allowed to stand at 60 - 70 ° c . for 48 h under a nitrogen atmosphere . the reaction mixture was washed three times each with ch 2 cl 2 , ch 3 oh , dmf , ch 2 cl 2 and ethyl acetate and the resultant resin was dried in vacuo . an aliquot of the resins was cleaved with 5 n naoh in ch 3 oh — ch 2 cl 2 ( 1 : 2 ) to afford a mixture of e - and z - isomers . the mixture was separated by chromatography on the silica gel to give a 23 % ( 0 . 13 mmol g − 1 ) yield of products , consisting of 21 % ( 0 . 12 mmol g − 1 ) of the e - isomer and 2 % ( 0 . 01 mmol g − 1 ) of the z - isomer . r f ( z - isomer = 0 . 58 ( hexane - ethyl acetate , 4 : 1 ); rf ( e - isomer )= 0 . 44 ( hexane - ethyl acetate , 4 : 1 ); amorphous ; 1 h nmr ( cdcl 3 , 300 mhz , δ ), 0 . 88 ( s , 3h , c 18 - methyl - h ), 1 . 2 - 2 . 4 ( m , steroid envelope and tributyl - stannyl - h ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 6 . 06 ( d , 1h , j = 19 . 4 hz , c 21 vinyl - h ), 6 . 22 ( d , 1h , j = 19 . 4 hz , c 20 vinyl - h ), 6 . 79 ( d , 1h , j = 2 . 4 hz , c 4 — h ), 6 . 84 ( dd , 1h , j = 2 . 6 , 8 . 4 hz , c 2 — h ), 7 . 28 ( d , 1h , j = 8 . 8 hz , c 1 — h ); 13 c nmr ( cdcl 3 ), 9 . 6 ( c 22 , 4c ), 13 . 7 ( c 24 , 4c ), 14 . 2 ( c 18 ), 23 . 4 ( c 15 ), 26 . 4 ( c 11 ), 27 . 3 ( c 25 , 4c ), 27 . 4 ( c 7 ), 29 . 2 ( c 23 , 4c ), 29 . 6 ( c 6 ), 32 . 4 ( c 12 ), 35 . 9 ( c 16 ), 39 . 4 ( c 8 ), 43 . 8 ( c 9 ), 46 . 7 ( c 13 ), 49 . 0 ( c 14 ), 85 . 6 ( c 17 ), 112 . 6 ( c 2 ), 115 . 2 ( c 4 ), 124 . 6 ( c 21 ), 126 . 5 ( c 1 ), 132 . 7 ( c 10 ), 138 . 3 ( c 5 ), 152 . 4 ( c 20 ), 153 . 3 ( c 3 ), ft - ir ( kbr ) v : 3445 ( 17β - oh , broad , 1719 ( c ═ o ), 1653 ( c ═ c ), 1607 , 1493 , 1451 ( aromatic ring ), 1217 ( c — o ). ( method b ). the 17α - ethynyl estradiol ( 3 g , 10 mmol ) was dissolved in thf and treated with triethylborane ( 2 ml , 17 mmol ) and tributyltin hydride ( 3 g , 11 mmol ). the mixture was stirred with a magnetic stirrer at 60 ° c . for 16 h . the crude mixture ( 7 . 73 g ) was evaporated to dryness , redissolved in the ch 2 cl 2 , and transferred to the swelled resin ( 5 g ) in ch 2 cl 2 in the presence of dcc . a catalytic amount of dmap was added to the mixture , which was allowed to stand for 24 h . the resultant functionalized resin was treated as previously described . the total loading for both e - and z - isomers was 0 . 59 mmol g − 1 with 0 . 56 mmol g − 1 of e - isomer and 0 . 03 mmol g − 1 of z - isomer , however , by the dry weight difference between pre - and post - reaction , the loading for both e - and z - isomers was 1 . 55 mmol g − 1 . the stille reaction was used to couple the anchored e - and z - stannylvinyl estradiol to aryl halides . the resin was added to the reaction flask , swelled in the ch 2 cl 2 , subsequently treated with 10 ml of anhydrous toulene . to the resultant slurry was added a 3 - 4 fold excess of the functionalized aryl halide , 1 - 2 crystals of 3 . 5 - di - t - butyl - 4 - hydroxytoulene ( bht ), and pd ( pph 3 ) 4 ( bowden , 1946 ; farrall , 1976 ). the reaction was allowed to proceed at 90 - 100 ° c . for 24 h . after cooling , the resin was washed as previously described , dried in vacuo and weighed . the resin was swelled in ch 2 cl 2 ( 10 ml ) containing 3 ml of 5 n - naoh in ch 3 oh - dioxane ( 1 : 3 ) and stirred for 1 h . this cleavage step was repeated three times . most of the product was collected from the first attempt , a small amount by second hydrolysis , and almost none from the third trial . the fractions were combined , evaporated to dryness , and partitioned between ethyl acetate and water . acetic acid ( 1 ml , 5 %) was added . the organic phase was washed with 10 % aqueous nahco 3 to remove the residual acetic acid , dried over mgso 4 , filtered and evaporated to dryness . the crude product was purified by silica gel column chromatography or by recrystallization from the appropriate solvent . 17α - 20e - 21 -( 2 - trifluoromethylphenyl )- 19 - norpregna - 1 , 3 , 5 ( 10 ), 20 - tetraene - 3 , 17β - diol ( 17α - e -( 2 - trifluoromethylphenyl )- vinyl estradiol ) ( 4a ). yield = 38 %; r t = 0 . 19 ( hexane - ethyl acetate , 4 : 1 ); mp 224 - 225 ° c . ; 1 h nmr ( 300 mhz , acetone - d 6 , 6 ) 1 . 02 ( s , 3h , c 18 methyl - h ), 1 . 2 - 2 . 4 ( m , steroid envelope ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 3 . 98 ( s , 1h , 17p hydroxyl - h ), 6 . 53 ( d , 1h , j = 2 . 3 hz , c 4 — h ), 6 . 58 ( dd , 1h , j = 2 . 6 , 8 . 5 hz , c 2 — h ), 6 . 64 ( d , 1h , j = 15 . 7 hz , c 20 vinyl - h ), 7 . 0 ( dd , 1h , j = 2 . 5 , 15 . 8 hz , c 21 vinyl - h ), 7 . 07 ( d , 1h , j = 8 . 7 hz , c 1 — h ), 7 . 42 ( t , 1h , j = 7 . 8 hz , c 26 — h ), 7 . 60 ( t , 1h , j = 7 . 3 hz , c 25 — h ), 7 . 69 ( d , 1h , j = 7 . 8 hz , c 27 — h ), 7 . 81 ( d , 1h , j = 8 . 3 hz , c 24 — h ), 7 . 98 ( s , c 3 hydroxy - h ); 13 c nmr ( 75 . 4 mhz , acetone - d 6 , 8 ) 14 . 7 ( c 18 ), 24 . 1 ( c 15 ) 27 . 2 ( c 11 ), 28 . 3 ( c 7 ), ( c 6 ), 33 . 4 ( c 12 ), 37 . 5 ( c 16 ), 40 . 7 ( c 8 ); 44 . 6 ( c 9 ), 48 . 4 ( c 13 ), 50 . 0 ( c 14 ), 84 . 3 ( c 17 ), 113 . 5 ( c 2 ), 115 . 9 ( c 4 ), 123 . 4 ( c 21 ), 125 . 6 ( q , j = 273 . 2 hz , c 28 : cf 3 ), 126 . 4 ( q , j = 5 . 8 hz , c 24 ), 127 . 0 ( c 1 ), 127 . 4 ( q , j = 29 . 4 hz , c 23 ), 127 . 8 ( c 26 ), 128 . 6 ( c 27 ), 132 . 0 ( c 25 ), 133 . 2 ( c 10 ), 137 . 9 ( c 22 ), 139 . 1 ( c 5 ), 142 . 4 ( c 20 ), 155 . 9 ( c 3 ); anal . calcd for c 27 h 29 o 2 f 3 : c , 73 . 30 ; h , 6 . 56 . found : c , 73 . 04 ; h , 6 . 68 . [ 0111 ] 17 α - 20e - 21 -( 3 - trifluoromethylphenyl )- 19 - norpregna - 1 , 3 , 5 ( 10 ), 20 - tetraene - 3 , 17β - diol ( 17α - e -( 3 - trifluoro methylphenyl )- vinyl estradiol ) ( 5a ). yield = 33 %; r f ( e - isomer )= 0 . 19 ( hexane - ethyl acetate , 4 : 1 ); mp 244 - 246 ° c . ; 1 h nmr ( 300 mhz , acetone - d 6 , 6 ), 1 . 01 ( s , 3h , c 18 - methyl ), 1 . 2 - 2 . 4 ( m , steroid envelope ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 3 . 98 ( s , 1h , 17β hydroxyl - h ), 6 . 53 ( d , 1h , j = 2 . 6 hz , c 4 — h ), 6 . 58 ( dd , 1h , j = 2 . 6 , 8 . 3 hz , c 2 — h ), 6 . 74 ( d , 1h , j = 16 hz , c 21 vinyl - h ), 6 . 84 ( d , 1h , j = 16 hz , c 20 vinyl - h ), 7 . 06 ( d , 1h , j = 8 . 3 hz , c 1 — h ), 7 . 54 - 7 . 56 ( m , 2h , c 25 , c 27 — h ), 7 . 75 - 7 . 79 ( m , 2h , c 23 , c 26 — h ), 7 . 93 ( s , c 3 - hydroxy - h ); 13 c nmr ( 75 . 4 mhz , acetone - d 6 , δ ), 14 . 7 ( c 18 ), 24 . 1 ( c 15 ), 27 . 3 ( c 11 ), 28 . 3 ( c 7 ), ( c 6 ), 33 . 5 ( c 12 ), 37 . 5 ( c 16 ), 40 . 7 ( c 8 ), 44 . 6 ( c 9 ), 48 . 4 ( c 13 ), 50 . 1 ( c 14 ), 84 . 2 ( c 17 ), 113 . 5 ( c 2 ), 115 . 9 ( c 4 ), 123 . 6 ( q , j = 5 . 6 hz , c 25 ), 124 . 1 ( q , j = 3 . 7 hz , c 23 ), 125 . 4 ( q , j = 271 hz , c 28 : cf 3 ), 126 . 0 ( c 26 ), 127 . 0 ( c 1 ), 130 . 2 ( c 21 ), 130 . 7 ( c 27 ), 131 . 2 ( q , j = 32 hz , c 24 ), 132 . 0 ( c 10 ), 138 . 4 ( c 5 ), 139 . 7 ( c 20 ), 139 . 9 ( c 22 ), 155 . 9 ( c 3 ); anal . calcd for c 27 h 29 o 2 f 3 : c , 73 . 30 ; h , 6 . 56 . found : c , 73 . 42 ; h , 6 . 68 . 17α - 20e - 21 -( 4 - trifluoromethylphenyl )- 19 - norpregna - 1 , 3 , 5 ( 10 ), 20 - tetraene - 3 , 17β - diol ( 17α - e -( 4 - trifluoro methylphenyl )- vinyl estradiol ) ( 6a ). yield = 49 %; r f = 0 . 15 ( hexane - ethyl acetate , 4 : 1 ); mp 215 - 217 ° c . ; 1 h nmr ( acetone - d 6 , 300 mhz , 8 ), 1 . 02 ( s , 3h , c 18 methyl - h ), 1 . 2 - 2 . 4 ( m , steroid envelope ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 3 . 90 ( s , 1h , 17β hydroxyl - h ), 6 . 53 ( d , 1h , j = 2 . 6 hz , c 4 — h ), 6 . 58 ( dd , 1h , j = 2 . 6 , 8 . 4 hz , c 2 — h ), 6 . 73 ( d , 1h , j = 16 hz , c 21 vinyl - h ), 6 . 85 ( d , 1h , j = 16 hz , c 20 vinyl - h , 7 . 07 ( d , 1h , j = 8 . 3 hz , c 1 — h ), 7 . 64 ( d , 2h , j = 8 . 7 hz , c 23 , c 27 — h ), 7 . 70 ( d , 2h , j = 8 . 6 hz , c 24 , c 26 — h ), 8 . 0 ( s , c 3 - hydroxy - h ); 13 c nmr ( 75 . 4 mhz , acetone - d 6 , 6 ) 14 . 7 ( c 18 ), 24 . 1 ( c 15 ), 27 . 3 ( c 11 ), 28 . 3 ( c 7 ), ( c 6 ), 33 . 5 ( cl 2 ), 37 . 6 ( c 16 ), 40 . 7 ( c 8 ), 44 . 6 ( c 9 ), 48 . 5 ( c 13 ), 50 . 2 ( cl 4 ), 84 . 2 ( c 17 ), 113 . 5 ( c 2 ), 115 . 9 ( c 4 ), 125 . 4 ( q , j = 270 . 6 hz , c 28 : cf 3 ), 126 . 0 ( c 21 ), 126 . 2 ( q , j = 3 . 5 hz , c 26 ), 126 . 2 ( q , j = 3 . 5 hz , c 24 ), 127 . 0 ( c 1 ), 127 . 6 ( c 23 , c 27 ), 128 . 9 ( q , j = 32 hz , c 25 ), 132 . 0 ( c 10 ), 138 . 4 ( c 5 ), 140 . 6 ( c 20 ), 142 . 7 ( c 22 ), 155 . 9 ( c3 ); anal . calcd for c 27 h 29 o 2 f 3 : c , 73 . 30 ; h , 6 . 56 . found : c , 73 . 36 ; h , 6 . 79 . 17α - 20z - 21 -( 4 - trifluoromethylphenyl )- 19 - norpregna - 1 , 3 , 5 ( 10 ), 20 - tetraene - 3 , 17α - diol ( 17α - z -( 4 - trifluoro methylphenyl )- vinyl estradiol ) ( 6b ). yield = 17 %; r f = 0 . 29 ( hexane - ethyl acetate , 4 : 1 ); 1 h nmr ( 300 mhz , acetone - d 6 , 6 ) 0 . 97 ( s , 3h , c 18 methyl - h ), 1 . 2 - 2 . 4 ( m , steroid envelope ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 3 . 89 ( s , 1h , 17p hydroxyl - h ), 6 . 12 ( d , 1h , j = 12 . 9 hz , c 21 vinyl - h ), 6 . 48 - 6 . 62 ( m , 3h , c 2 , c 4 , c 20 vinyl - h ), 7 . 11 ( d , 1h , j = 8 . 1 hz , c 1 — h ), 7 . 59 ( d , 2h , j = 8 . 4 hz , c 23 , c 27 — h ), 7 . 80 ( d , 2h , j = 8 . 4 hz , c 24 , c 26 — h ), 7 . 95 ( s , c 3 hydroxy - h ). 17α - 20e - 21 -( 2 - methylphenyl )- 19 - norpregna - 1 , 3 , 5 ( 10 ), 20 - tetraene - 3 , 17β - diol ( 17α - e -( 2 - methylphenyl )- vinyl estradiol ) ( 7a ). yield = 38 %; r t = 0 . 18 ( hexane - acetone , 4 : 1 ); mp 199 - 200 ° c . ; 1 h nmr ( acetone - d 6 , 300 mhz , δ ), 1 . 01 ( s , 3h , c 18 methyl - h ), 1 . 2 - 2 . 4 ( steroid envelope ), 2 . 34 ( s , 3h , c 28 methyl - h ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 3 . 84 ( s , 1h , 17β hydroxyl - h ), 6 . 44 ( d , 1h , j = 16 hz , c 21 vinyl - h ), 6 . 52 - 6 . 63 ( m , 2h , c 2 , c 4 — h ), 6 . 83 ( d , 1h , j = 16 hz , c 20 vinyl - h ), 7 . 07 ( d , 1h , j = 8 . 3 hz , c 1 — h ), 7 . 10 - 7 . 15 ( m , 3h , c 24 , c 25 , c 26 — h ), 7 . 48 ( d , 1h , j = 6 . 8 hz , c 27 — h ), 7 . 97 ( s , c 3 hydroxy - h ); 13 c nmr ( 75 . 4 mhz , acetone - d 6 , δ ) 14 . 7 ( c 18 ), 19 . 9 ( c 28 , methyl ), 24 . 1 ( c 15 ), 27 . 3 ( c 11 ), 28 . 3 ( c 7 ), ( c 6 ), 33 . 5 ( c 12 ), 37 . 5 ( c 16 ), 40 . 7 ( c 8 ), 44 . 7 ( cg ), 48 . 2 ( c 13 ), 50 . 1 ( c 14 ), 84 . 2 ( c 17 ), 113 . 5 ( c 2 ), 115 . 9 ( c 4 ), 125 . 4 ( c 26 ), 126 . 5 ( c 25 ), 126 . 9 ( c 24 ), 127 . 0 ( c 1 ), 127 . 7 ( c 21 ), 130 . 8 ( c 27 ), 132 . 0 ( c 10 ), 135 . 9 ( c 20 ), 137 . 9 ( c 22 ), 138 . 4 ( c 5 ), 138 . 8 ( c 23 ), 155 . 9 ( c 3 ); anal . calcd for c 27 h 32 o 2 : c , 83 . 51 ; h , 8 . 25 . found : c , 83 . 79 ; h , 8 . 65 . 17α - 20e - 21 -( 3 - methylphenyl )- 19 - nonpregna - 1 , 3 , 5 ( 10 ), 20 - tetraene - 3 , 17β - diol ( 17α - e -( 3 - methylphenyl )- vinyl estradiol ) ( 8a ). yield = 75 %; r t = 0 . 17 ( hexane - acetone , 4 : 1 ); mp 204 - 205 ° c . ; 1 h nmr ( 300 mhz , acetone - d 6 , 6 ), 1 . 00 ( s , 3h , c 18 methyl - h ), 1 . 2 - 2 . 4 ( m , steroid envelope ), 2 . 31 ( s , 3h , c 28 methyl - h ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 3 . 74 ( s , 1h , 17β hydroxyl - h ), 6 . 52 - 6 . 63 ( m , 4h , c 4 , c 2 , c 21 vinyl , c 20 vinyl - h ), 7 . 03 ( d , 1h , j = 7 . 3 hz , c 25 — h )., 7 . 07 ( d , 1h , j = 8 . 7 hz , c 1 — h ), 7 . 16 - 7 . 31 ( m , 3h , j = 7 . 4 hz , c 23 , c 26 , c 27 — h ), 7 . 93 ( s , 1h , c 3 hydroxy - h ); 13 c nmr ( 75 . 4 mhz , acetone - d 6 , 6 ) 14 . 8 ( c 18 ), 21 . 4 ( c 28 ; methyl ), 24 . 1 ( c 15 ), 27 . 3 ( c 11 ), 28 . 4 ( c 7 ), ( c 6 ), 33 . 5 ( c 12 ), 37 . 4 ( c 16 ), 40 . 8 ( c 8 ), 44 . 7 ( cg ), 48 . 3 ( c 13 ), 50 . 2 ( c 14 ), 84 . 2 ( c 17 ), 113 . 6 ( c 2 ), 116 . 0 ( c 4 ), 124 . 4 ( c 27 ), 127 . 0 ( cl ), 127 . 7 ( c 25 ), 127 . 8 ( c 26 ), 128 . 5 ( c 21 ), 129 . 2 ( c 23 ), 132 . 2 ( c 10 ), 137 . 0 ( c 20 ), 138 . 4 ( c 5 ), 138 . 7 ( c 22 , c 24 ), 155 . 9 ( c 3 ); anal . calcd for c 27 h 32 o 2 : c , 83 . 51 ; h , 8 . 25 . found : c , 83 . 23 ; h , 8 . 42 . 17α - 20z - 21 -( 3 - methylphenyl )- 19 - norpregna - 1 , 3 , 5 ( 10 ), 20 - tetraene - 3 , 17β - diol ( 17α - z -( 3 - methylphenyl )- vinyl estradiol ) ( 8b ). yield = 54 % ( 0 . 01 g ); r t = 0 . 25 ( hexane - acetone , 4 : 1 ); 1 h nmr ( 300 mhz , acetone - d 6 , δ ) 0 . 95 ( s , 3h , c 18 methyl - h ), 1 . 2 - 2 . 4 ( m , steroid envelope ), 2 . 31 ( s , 3h , c 28 methyl - h ), 2 . 7 - 2 . 9 ( m , 2h , c 6 — h ), 3 . 27 ( s , 1h , 17p hydroxyl - h ), 5 . 96 ( d , 1h , j = 13 . 1 hz , c 21 vinyl - h , 6 . 44 ( d , 1h , j = 13 . 1 hz , c 20 vinyl - h ), 6 . 53 ( d , 1h , j = 2 . 6 hz c 4 — h ), 6 . 60 ( dd , 1h , j = 2 . 6 , 8 . 3 hz , c 2 — h ), 7 . 03 ( d , 1h , j = 7 . 3 hz , c 25 — h ), 7 . 11 ( d , 1h , j = 8 . 3 hz , c 1 — h ), 7 . 17 ( t , 1h , j = 7 . 6 hz , c 26 — h ), 7 . 38 - 7 . 43 ( m , 2h , c 23 , c 27 — h ), 7 . 95 ( s , 1h , c 3 hydroxy - h ); 13 c nmr ( 75 . 4 mhz , acetone - d 6 , 6 ) 14 . 58 ( c 18 ), 21 . 42 ( c 28 : methyl ). 23 . 85 ( c 15 ), 27 . 40 ( c 11 ), 28 . 30 ( c 7 ), ( c 6 ), 32 . 97 ( cl 2 ), 38 . 4 ( c 16 ), 40 . 9 ( c 8 ), 44 . 7 ( cg ), 48 . 8 ( c 13 ), 50 . 1 ( c 14 ), 84 . 3 ( c 17 ), 113 . 6 ( c 2 ), 116 . 0 ( c 4 ), 127 . 1 ( c 1 ), 127 . 8 ( c 27 ), 128 . 1 ( c25 ), 128 . 3 ( c 26 ), 129 . 7 ( c 21 ), 131 . 4 ( c 23 ), 132 . 0 ( c 10 ), 137 . 1 ( c 20 ), 137 . 6 ( c 24 ), 138 . 45 ( c 5 ), 138 . 5 ( c 22 ), 155 . 9 ( c 3 ); anal . calcd for c 29 h 36 o 3 : c , 80 . 55 ; h , 8 . 33 . found : c , 80 . 00 ; h , 8 . 41 17α - 20e - 21 ( 4 - methoxyphenyl )- 19 - norpregna - 1 , 3 , 5 ,( 10 ), 20 - tetraene - 3 , 17β - diol ( 17α - e -( 4 - methoxyphenyl )- vinyl estradiol ) ( 9a ). yield = 36 %; r t = 0 . 23 ( chcl 3 — ch 3 oh , 99 : 1 ); 1 h nmr ( 300 mhz , acetone - d 6 , δ ) 0 . 99 ( s , 3h , c 18 methyl - h ), 3 . 68 ( s , 1h , 17p hydroxy - h ), 3 . 78 ( s , 3h , c 28 : methoxy - h ), 6 . 46 ( d , 1h , j = 16 . 1 hz , c 21 — h ), 6 . 51 - 6 . 59 ( m , 3h , c 2 , c 4 , c 20 — h ), 6 . 88 ( d , 2h , j = 8 . 8 hz , c 24 , c 26 — h ); 7 . 07 ( d , 1h , j = 8 . 3 hz , cl - h ); 7 . 39 ( d , 2h , j = 8 . 8 hz , c 23 , c 27 — h ), 7 . 95 ( s , 1h , c 3 hydroxy - h ); 13 c nmr ( 75 . 4 mhz , acetone - d 6 , δ ) 14 . 7 ( c 18 ), 24 . 1 ( c 15 ), 27 . 3 ( c 11 ), 28 . 3 ( c 7 ), ( c 6 ), 33 . 4 ( c 12 ), 37 . 3 ( c 16 ), 40 . 7 ( c 8 ), 44 . 7 ( cg ), 48 . 2 ( c 13 ), 50 . 0 ( c 14 ), 55 . 5 ( c 28 : methoxy ), 84 . 1 ( c 17 ), 113 . 5 ( c 2 ), 114 . 7 ( c 24 , c 26 ), 115 . 9 ( c 4 ), 127 . 0 ( c 1 ), 127 . 0 ( c 21 ), 128 . 3 ( c 23 , c 27 ), 131 . 4 ( c 22 ), 132 . 1 ( c10 ), 134 . 9 ( c 20 ), 138 . 4 ( c 5 ), 155 . 9 ( c 3 ), 159 . 9 ( c 25 ). in the above procedures , the solid phase synthesis methodology was applied using carboxylated resins to generate a series of novel er - lbd ligands , or estradiol derivatives . the purification steps were simplified and simultaneously produced both the e - and z - isomers . yield may be improved by modifications in both the coupling and cleavage steps for a chemically more sensitive z - isomer . one of the key elements of the synthetic scheme was the selection of a linker that could be both formed and cleaved under mild conditions . 17α - substituted estradiols were unstable under strongly acidic conditions such as those frequently used to release products from the resins . therefore the resin of choice was carboxylated polystyrene which could be esterified under neutral conditions and ultimately cleaved with mild base . compound 8a was prepared using the carboxylated resin obtained either by oxidation of a wang resin using jones reagent ( bowden , 1946 ) or by carboxylation of a polystyrene resin via lithiation with n - butyl lithium ( farrall , 1976 ). the reactions for both methods were easily monitored by the appearance of the 1700 cm − 1 absorption in the ft - ir spectrum . the loading capacity of the carboxylated resins was determined by coupling 17α - ethynyl estradiol onto the resins using dcc in the presence of catalytic amount of dmap and measuring its subsequently cleaved estradiol derivatives from the aliquot of the resins . the loading 1 of oxidized wang resin was 0 . 4 - 0 . 6 mmol g − 1 and that of carboxylated polystyrene was 1 . 5 - 1 . 9 mmol g − 1 . once the utility of coupling through the ester linkage using carboxy polystyrene resin was confirmed , the commercially available carboxy poolystyrene was used for the remainder of the work . the loading yield of the reaction using the resins with already known loading capacity ( 2 . 47 mmol g − 1 ) was 82 %. the yield was determined by ‘ cleave and characterize ’ methods . as shown in fig1 the synthesis of the analogs was initiated by coupling the 3 - phenolic group of 17α - ethynyl estradiol to the carboxy polystyrene resin . the steroids on the resins were confirmed by an antimony ( iii ) chloride assay ( carr , 1926 ; blatz , 1972 ; jork , 1990 ). due to the absence of color change with bromocresol green , no free carboxylic acid groups remained on the resin ( gordon , 1972 ). the appearance of a peak at 3301 cm − 1 in the ir spectrum , corresponding to the c — h stretch of the ethynyl group , also confirmed that the reaction and a shift of carbonyl absorption to higher frequency ( from 1690 - 1734 cm − 1 ) was also observed . the subsequent hydrostannylation step incorporated either the use of hydrostannylation of bound ethynyl estradiol ( method a ) or hydrostannylation of ethynyl estradiol in solution phase synthesis followed by coupling to the resin ( method b ). the resin - bound 17α - ethynyl estradiol was hydrostannylated with tributyltin hydride using triethyl - borane as a radical initiator ( nozaki , 1989 ) to afford a mixture of the 17α - e / z - tri - n - butylstannylvinyl estradiol in 20 - 30 % ( 0 . 12 mmol g − 1 of e , 0 . 01 mmol g - 1 of z ) loading yields . varying the reaction conditons , e . g . different solvents , temperatures , or reaction times , did not improve the yields . therefore , a direct coupling of 17α - e / z - tri - n - butylstannyl - vinyl estradiols was used to overcome the low efficiency of this step . 17α - ethynyl estradiol was hydrostannylated to 60 ° c . and the crude mixture was directly transferred to the resin slurry in ch 2 cl 2 . the mixture was treated with a 2 - 3 fold excess of dcc and a catalytic amount of dmap was added . the loading yield for the coupling reaction was 0 . 59 mmol g − 1 with a z / e ratio = 1 : 20 . the low loading yield was due to the use of the acetic acid for the protonation of phenoxide ion after cleavage , subjecting the products to protiodestannylation and reducing the expected loading yield . because the cleavage after hydrostannylation did not provide a precise loading yield , the dry weight difference between pre - and post - reaction was subsequently used to determine the loading yield . using the dry weight difference method , the yield for the hydrostannylation reaction was 1 . 55 mmol g − 1 for both e - and z - isomers . because hydrostannylation on the resin did not afford satisfactory yields . method b was the method of choice . the ratio of e and z isomers is a function of the reaction temperature , time and stoichiometric ratio of tributyltin hydride to alkyne . at 60 ° c . the reaction generated greater than 20 : 1 ( e / z ) ratio bound to the solid phase . to increase the ratio of the z - isomer , triethylborane was used as a radical initiator and the reaction was run at low temperature . the proportion of the z - isomer ( z / e = 1 : 10 ) was increased . however , the reaction required a longer time and the loading yield for the hydrostannylation was slightly lower than at higher temperature ( 1 . 44 mmol g − 1 by the dry weight difference method ) because of more unreacted 17α - ethynyl estradiol in the reaction mixture . the resin - bound hydrostannnylated estradiol was subjected to the stille coupling reaction ( stille , 1985 ) using a variety of substituted aryl halides to generate the target compounds ( see table 2 ). as shown in fig1 pd ( pph 3 ) 4 was used as the catalyst for the reaction and 3 , 5 - di - t - butyl - 4 - hydroxytoluene ( bht ) was added as a scavenger . the use of pd ( pph 3 ) 4 generated an insoluble by - product that caused coloration of the resin , however , it was easily removed by rinsing it through the built - in filter ( 50 - 70 μm ). after completion of all the reaction steps , the product was cleaved from the resin by saponification with 5 n naoh dissolved in ch 3 oh - dioxane ( 1 : 3 ). as shown in table 2 , the unoptimized yields of the stille reactions on solid phase ranged from 17 - 75 %, comparable to those observed for solution phase synthesis . compounds 5a ( para - trifluoromethylphenyl , e - isomer ) and 5b ( para - trifluoromethylphenyl , z - isomer ) were isolated from the stille reaction in a ratio of 98 : 2 . compound 7a ( meta - methylphenyl , e - isomer ) and 7b ( meta - methylphenyl , z - isomer ) were also obtained in a ratio of 96 : 4 . although the z - tri - n - butylstannyl vinyl estradiol was initially present on the resin , no z - isomers of compounds 3a , 4a , 6a , or 8a were isolated from the stille coupling , instead , 17α - vinyl estradiol , resulting from protiodestannylation was recovered as a side product . because an excess of reagent was used to drive the reaction to completion , unreacted hydrostannylated 17α - e / z -( tri - n - butylstannyl )- vinyl estradiol was no detected after the stille reaction . it is possible that the z - isomers either isomerized to thermodynamically more stable e - isomers under the conditions required for the stille reaction or underwent protiodestannylation . as previously observed , the z - isomer is much more susceptible to protiodestannylation than the e - isomer and the appearance of the side product under either solid phase or solution phase synthesis was approximately the same . the isolated product were characterized by standard spectroscopic methods ( ft - ir , 1 h and 13 c nmr ) and analytical methods . the data were consistent with the proposed structures . stereochemical assignments for compounds 5a and 5b were based on the c 20 , c 21 olefinic proton coupling constants for which e = 16 hz and z - 12 . 9 hz , respectively . for compounds 7a and 7b , the observed coupling constants were 18 . 2 hz of the c 20 e - vinyl proton and 13 . 1 hz for the c 20 z - vinyl proton . in 13 c nmr , long range couplings were observed for the compounds 3a - 5a and 5b containing the trifluoromethyl group . coupling with strongly electronegative fluorine was found at the carbon directly attached to the fluorine ( 1j c - f ) and one ( 2 j c - f ) and two carbons distant ( 3 j c - f ). the carbons appeared as quartets and the coupling constants were approximately 1 j c - f = 270 hz . 2 j c - f = 32 hz . 3 j c - f = 3 - 5 hz respectively . ortho , meta , and para ( trifluoromethyl ) phenylvinyl estradiol isomers were evaluated for estrogen receptor - ligand binding domain ( er - lbd ) affinity . the properties of the aryl substituent and its position on the ring ( ortho / meta / para ) affect receptor binding . trifluoromethyl group was introduced onto phenylvinyl estradiol either at the ortho , meta , or para positions . these compounds were examined for their ability to stimulate or inhibit estrogen responses in two assay systems . the initial system evaluated the ability of the ligand to stimulate the proliferation of mcf - 7 cells and as the results in fig1 indicate , the ortho - isomer produced a full agonist response comparable to that of estradiol . when the ligand was added to the cells in the presence of 1 nm estradiol , there was neither an enhancement nor a diminution of the proliferative response . the meta - and para - isomers gave substantially different profiles . the meta - isomer demonstrated a weak proliferative effect at doses greater than 1 nm and antagonized the effects of estradiol at the same doses . the para - isomer , however , did not elicit a proliferative response until a 10 nm dose was employed and decreases in the estradiol effects were observed below 1 nm . therefore , the position of the trifluoromethyl group exerted a significant effect on the efficacy of the ligand . these trifluoromethyl substituted compounds were also studied with an immature female rat uterotrophic growth assay ; the results are shown in fig1 , 13 , and 14 . in the estrogenic assay , the ortho - isomer produced an effect comparable to estradiol at a 3 nm level and substantial estrogenic effects at 10 and 100 nm . the meta - and para - isomers , however , demonstrated little or no estrogenic effects , even at 10 and 100 nm . therefore , the agonist responses observed in the in vitro cell proliferation assay were carried over to the intact animal as well . the antiestrogen assay evaluated the ability of the isomers to block the uterotrophic effect induced by 1 nm estradiol . under these conditions , the ortho isomer produced an enhancement of the estrogenic response at both 10 and 100 nm . the meta - isomer demonstrated no significant effect on the estradiol response at either dose , however , the para isomer reduced the estrogenic response at the 100 nm level . therefore , in both estrogen responsive cells and tissues these new ligands are producing differential responses in affinity and efficacy related to the site of trifluoromethyl substitution on the phenyl ring . the cellular target for antiandrogen therapy , the androgen receptor ( ar ), is a member of the nuclear receptor superfamily which has been studied extensively over the past decade ( tsai , 1994 ). members of this receptor bear a strong structural similarity ( homology ) and utilize similar signaling pathways to express their biological actions . at the molecular level , the ar , like the other steroid hormone receptors , is composed of discrete domains that are responsible for specific functions . the hormone binding domain ( hbd ), the sequence of aminoacids near the n - terminus of the ar , recognizes and binds testosterone with high affinity but not other hormones or small endogenous molecules ( weatherman , 1999 ; simons , 1998 ). this region of the receptor has been examined using x - ray crystallography to elucidate the aminoacid residues responsible for the recognition of specific hormones . the hormone binding domains on the estrogen receptor ( er ), progesterone receptor ( pgr ) and retinoic acid receptor ( rar ) provide a common fold for the endogenous hormone , which also strongly suggest the types of conformational changes that occur upon ligand binding ( brzozowski , 1997 ; tannenbaum , 1998 ; shiau , 1998 ; williams , 1998 ; renaud , 1995 ; klaholz , 1998 ). the conformational changes , particularly those associated with helix - 12 , assist in the recruitment of specific coactivator proteins that appear to initiate the action of the general transcription apparatus ( resche - rigon , 1998 ; mckenna , 1999 ; klinge , 2000 ). in accordance with the present invention , the steroidal nucleus is the address component , which directs the molecule to the hbd where , for agonists , the d - ring substituents direct helix - 12 into a conformation that exposes the activation function - 2 ( af - 2 ) or message component . for known er and pgr antagonists , the steroid nucleus present in most drugs provides the appropriate address . however , the incorporation of an additional functional group interferes with the movement of helix - 12 , and produces a full or partial antagonist response ( message ). most of the antihormones known in the art incorporate that additional functional group at either the 11β - or 7α - position of the steroid ( see fig7 ). the present invention shows that antagonism can be generated through introduction of an appropriate 17α - substituent . significant research efforts have focused on the synthesis and evaluation of compounds designed to either mimic or block the effects of the endogenous androgen , testosterone . while many steroidal compounds can mimic testosterone , relatively few were able to block its effects in target tissues and virtually none were effective in treating hormone responsive prostate cancer ( teutsch , 1995 ). nonsteroidal agents , however , such as ( hydroxy ) flutamide , nilutamide , and bicalutamide ( sciarra , 1990 ; tucker , 1988 , 1990 ), have demonstrated clinical efficacy for the treatment of prostatic carcinoma , even though their affinity for the ar is relatively low when compared to testosterone ( kokontis , 1999 ; battmann , 1998 ). recent publications have disclosed another class of nonsteroidal antiandrogens which have potential as clinically useful agents ( hamann , 1998 ; edwards , 1999 ; higuchi , 1999 ; kong , 2000 ). analogs of these compounds also demonstrate agonist / antagonist responses at other nuclear receptors ( pooley , 1998 ; zhi , 1998 , 1999 , 2000 ). because the nonsteroidal antiandrogens do not correspond to any current steroid hormone pharmacophore , it is possible that they may primarily effect only the message region ( helix - 12 ) of the ar - hbd . a potent interaction at that site would still compete with agonist ligand binding for the address region , not entirely unlike the situation for the dopamine transporter inhibitors where structurally diverse families of ligands not only inhibit dopamine and cocaine binding but also , by associating with overlapping sites , inhibit the binding of each other . thus , the present invention combines features from both the steroid nucleus ( address component ) and the nonsteroidal antagonist pharmacophore ( message component ) ( see fig8 ). a combination of organotin chemistry and palladium catalyzed coupling reactions is used for the synthesis of the message components ( see fig1 ). the 1 - ethynyl - 1 - aminoperhydroindanes which would incorporate the c - and d - rings of the steroid nucleus is prepared from the corresponding 1 - ethynyl - 1 - acetoxy analogs using a cu ( i )- assisted aminolysis . the ethynyl cycloalkyl alcohols or amines readily undergo hydrostannation to give the corresponding e - and z - stannylvinyl intermediates which can be coupled with the requisite mono - or di - substituted aryl iodide under stille coupling conditions ( farina , 1995 ; casado , 1998 ). three 3 ′- or 4 ′- substituted , three 3 ′-, 4 ′- disubstituted , and three 3 ′-, 5 ′- disubstituted phenyl iodides are used to generate a total of 18 compounds . while there are no obvious choices for the optimal substituents , the structure activity relationships ( sar ) for antiandrogens suggest that electron withdrawing groups ( e . g ., — no 2 , cf 3 ) enhance potency . therefore , these groups are used with one electron releasing group in the first series ( tucker , 1988 ). suzuki coupling reaction is used with vinylboronic acid ( suzuki , 1999 ). the e - vinylboronic acid is accessed directly by hydroboration of the alkyne with catecholborane followed by hydrolysis . the z - isomer is obtained from the z - vinylstannane via idododestannylation , followed by coupling with bispinacolatodiboron , and hydrolysis . for the synthesis of the spirocyclic ether or amine message components , the coupling partner for the z - vinylstannane ( or boronic acid ) requires an orthoiodo ( bromo ) phenol derivative . halogenation of the commercially available 3 ′- or 4 ′- substituted phenol gives the intermediate which is initially protected as the silyl ether . the z - vinyl arene is made by the standard stille or suzuki coupling methods . the conditions developed by buchwald and hartwig to effect the intramolecular aryl amine / ether formation may be used ( wolfe , 1998 , 1999 ; yang , 1999 ; hartwig , 1998a , b ) deprotection of the phenol , conversion to the triflate , and coupling with an appropriate pd catalyst , such as pd 2 ( dba ) 3 , and an activating ligand , such as binap , will effect the cyclization . the final product is provided by the deprotection of the amine . each new compound synthesized is characterized by the standard spectrometric methods — high resolution mass spectrometry ( hrms ), h - 1 / c - 13 - nuclear magnetic resonance spectrometry ( nmr ) to confirm the proposed molecular structures . solution conformations is determined by using 1d - and 2d - nmr techniques , methods of which are described above . the use of both conformational analysis and computational methods , more probable solution conformations are identified , which provides information with regard to key structural features and how they influence molecular conformations . compounds prepared containing the message components may be screened by a bioevaluation protocol already established through a commercially available company ( e . g ., mds - panlabs , located in bothell , wash .) to determine their ar affinity , efficacy and selectivity . receptors from rat ventral prostate tissue may be used to determine the ic50 and ki values . [ h - 3 ] mibolerone may be used as the radioligand . synthesized hydroxyflutamide , nilutamide , bicalutamide and lg 120907 is evaluated as standard ar ligands . those new compounds that demonstrate ar affinities & gt ; 10 % that of bicalutamide or lg 120907 will be evaluated for their affinities for the other nuclear receptors . other sources for receptors and their radioligands include erα - human recombinant from insect st9 cells , [ h - 3 ] estradiol , gr - human jerkat cells , [ h - 3 ] dexamethasone , and pgr - bovine , [ h - 3 ] r - 5020 . compounds that express a significant selectivity for ar (& gt ; 10 : 1 ) is tested for their efficacy in the rat agonism / antagonism model . in vitro efficacy model for testing the compounds for antagonism is the use of cotransfection and whole cell receptor binding ( hamann , 1998 ). preliminary sars is determined from the ic50 and ki data from the screening of the new compounds . e - vs . z - stereochemistry of the acyclic series of compounds is studied as well as the effects of mono - vs . di - substitution and 3 - vs . 4 - substitution . the cyclized compounds are compared with the acyclic series - to identify particular substituent trends . the qsar - comfa module of sybil is used to clarify the individual parameters ( gantchev , 1994 ). the physicochemical parameters developed by hansch may also be used to evaluate the data ( gantchev , 1994 ). the most potent ligands are analyzed for the lowest energy conformations using quanta - charmm / mm3 force fields ( wurtz , 1998 ) and compared with those from the nmr conformational studies to rationalize the initial sar . this allows for better determination of which substituents are most effective in contributing to ar affinity , selectivity and antihormonal response . subsequently , the selected substituents is used for incorporation into the address - message composite . 17α - ethynyl -( 19nor ) testoterone and its dihydroderivative ( address component ) is used as the starting material . the message components may be obtained from commercially available ( or readily synthesized ) mono - and disubstituted iodophenols . the same message components as with the estrogen study are used — the nilutamide / bicalutamide family of nonsteroidal antagonists and the more potent ligand pharmaceutical antagonists . for the message components analogous to flutamide and bicalutamide , the ethylene group is selected as an isosteric substitution for the amide bond ( luthman , 1996 ). the method for synthesis of the ( nor ) testosterone derivatives with the message component at the 17α - position is similar to the steps used for the synthesis of antiestrogens described herein . the antiandrogens of the present invention will include the steroid nucleus ( a - d rings ) and will provide functionality in the a - ring ( 3 c ═ o /— oh ; 4 , 5 - c ═ c ). as an embodiment , these groups are prepared to protect them as ketals , esters or silyl / enol ethers ( hoyte , 1993 ; van den bos , 1998 ). battmann , t ., branche , c ., borichoux , f ., cerede , e ., philibert , d ., goubet , f ., teutsch , g ., gallard ,- kelly , m . pharmacological profile of ru58642 , a potent systemic antiandrogen for the treatment of androgen - dependent disorders . j . steroid biochem . molec . biol . 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( 1999 ) 42 : 1466 - 1472 . while the present invention has been described in conjunction with a preferred embodiment , one of ordinary skill , after reading the foregoing specification , will be able to effect various changes , substitutions of equivalents , and other alterations to the compositions and approaches set forth herein . it is therefore intended that the protection granted by letters patent hereon be limited only by the definitions contained in the appended claims and equivalents thereof .