AU2008202236A1 - Selective androgen receptor modulators and methods of use thereof - Google Patents

Description

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AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION Standard Patent Applicant: University of Tennessee Research Foundation Invention Title: SELECTIVE ANDROGEN RECEPTOR MODULATORS AND METHODS OF USE

THEREOF

The following statement is a full description of this invention, including the best method for performing it known to me/us: P60525AUA. PalSetFling Application 2008-5-2.doc (8) 00 SNELBCnVE ANDROQEN IRECR"DTR MODULATORS AND METHODS OF USE THEREOF C~1 FIELDJ OF INVEl IOPN The present invention Tolates to a novel clans of tissue-selective androgen INDre"ePtor targeting agents (ARTA) which demonstrate androgtaic and anabolic activity.

of a nonstaroidal ligand for the androgent receptor. The agents deflne a now subclos, of C~~1 omipounds which ane tissue-seleotive androgen receptor mnodulators (SARM) which are useful for male hormone therapy such as oral testosterone replacement theray, male 00 contraception, maintaining. sexual desire in women, treating prostate cancer, ad imaging prostate cancer. Thao agents are also adini~stered to a subject for the treatment of sarcopenia, lacki of sexual libido, osteoporosis, ezythropoiesis, and fertility.

T1e agents way be used alone or in combination with 4 progestin or estrogen.

13ACKGROUNI) OF THE INVENTION The andirogen receptor (MR" is a ligand-aotivaWe transcriptional regulatory protein that mediates induction of male sexu~al development and Aljntion through its activity with endogenouis androgens. Androgens are generaly );nown as the nate sex hormones. However, androgens also play a pivotal role in female physiology and reproduction. The androgenic hormones ame steroids which are produced in the body by the testis and the cortex of the adrenal gland, or synthesized in the laboratory.

Androgenic steroids play an important role in many physiologic processes, including the development arnd malntenance of male sexual characteristics such as muscle and bone mass. prostate growth. spetmatogenesis, and the male hair pattern (Matsumoto, *Endoodinol. Met Clan. N. Am. 23:857-75 (:1994). The endogenous steroidal androgens include te-stostemoe and dibydrotestostaroue Testosterone is the principal sterid secreted by the testes and is the primary Circulating anidrogen found in the plasma of males. Testosterone is converted to DHT by the enzyme 5 alpha-reduotase in many peripheral tissu~es. DHT is thus thought to serve 4s the intracellular mediator for most androgen actions (Zhou, at al., Molec. Endocrinal. 9:208-12 (1995))- Other atervidal androgens include estrsr of testosterone, suab as the cypiovate, propionate.

phenylpropionate, cyclopeiitylpropionate, isocasporate, enanthate, and decanoate esters, and other synthetic androgens sach as 7-Mothyl-Nortestosterona ('"MTNM~ and Its 00 acetate ester (Sundawn et al., "7 Alphs-Methyl-Nortestostorone(MBAfl: The Optiml Andcrogen For Male Conzraception," Ann. Med., 25:199-205 (1993) ("Sundarain")).

SecWIS the AR is involved in male sexual development and finction, the AR is a likely target for efteting male contraception or other farins of borrmone replacement therapy.

The AR also regulates femnale sexual flinction libido), bone formation, and INDrooea~ c-i Worldwide. populationi growth and social awarenass of famnily planning have Stimulated a great deal of research in contraception. Contraception is a dtifficult subject 00 10 under any circumstances. It is fraught with cuihiral and social stigm, religious imnplications, and. most certainly, significant health concerns. This sitiation is only exacerbated when the subject focuses on male contraception. Despite the availability of suitable contraceptive devices, historically, society has looked to women to be res-ponsible for contraceptive decisions and their consequences. Although health concerns over sexually transmitted diseases have made men more aware of the need to develop safe and responsible sexual habits, women still often bear the brunt of contraceptive choice. Women have a number of choices, fin temporary maechanical devices such as sponges and diaphragms to temporary chemical devices such as spermicides. Women also have at their disposal more permanent options, such as physical devices Wik 1UDs end cervical caps as well as more permannt chernioal teaets, such as birth control pills and subcataneous implants. HIowever, to dafte, the only options available for men include the use of condoms or a vasectomy. Condom use, however is not favore5 by many men because of the reduced sexual sensitivity, the interution in sexual spontanety, and the signficant possibility of pregwacy caused by breakage or misuse. Vasectomies ame also not t~vored. If more convenient methods of birth control were *vatilable to meni, particularly long term methods that require no prepatrative activity Immediately prior to a sexual act such methods could significantly Increase the likeliood that men would take more responsibility for contracption.

Admninmtion of &ha male sex steroids testosterone and its derivatives) has shown patcular promise in tbis regard due to te combined gonadotropinsuppressing arid anclrogen-substituting properties of threse compotinds (Stainberger at al., 00 "Effect Of Chronic Administration of Testosterone En8thIate on Sperm Produotion and Plasmas Testo~sterone, Follicle SthmuLating Hormone, and tLuteinizng ?Lornone Levels.- A Prelimiary Evaluation of a Possible Male Contraceptive, Fertility and Sterility 29:1320- 28 (1977)). Chmdac administration of high doses of testosterone completely S abolishes sperm production (eaopern-da) or reduces it to a very low level (oligospermia). The degree of spermtogenic suppression necessary to produce infertility is not precisely known. However. a recent report by the World Health Organization showed that weekly intramuiscular injections of testogteronie ozianthate- result in azoospennia or severe oligospena toos than 3 million sperm per mut) and infertility 00 0 in 98% of men receiving therapy (World Health Organization Task Force on Methods Ar R~egulation of Male Fertility, "Contraceptive Efficacy of Testosterone-bIduced Azoosperinia and Oligospermia in *Normal Men," Fertifily and Sterility 65:821-29 (1996)).

A variety of testosterone est=r have been developed that arm more slowly absorbe after inuscular injection anid, tus, result in greater androgenic effect.

Testosterone enantte is the most widely used of these caters. While testosterone enanthate has been valuable in term of establishing the feasibility of hormonal agents for Male contraception, it has saveal. drawbacks, including the need for weekly injections and the presence of supraphysiologic. peak levels of testosterone immediatly following intramuscular iujection (Wu, "Bifeots of Testosterone Enanthate in Normal Men: Experience Prom a Multicenter Contrcetive Efficacoy Study," Fertily and Sterility 65:626-36 (1996)).

2S Steroidal ligmnds which. bind the AR and act as anidrogens testostemoe eztnthate) or as antiandrogens cyproterone acetate) have boon known for mny Yers and are used cliaically (Wu 1988). Although nonsteroldal antiandrogens are in clinlical use for hormone-dependent prostate cancer. nonsteridal androgens have not boon reported. For this meason. reseach on male contraceptives bas focused solely on steroidal comzpouTds.

00 C~1 SUKKARY OF THE INVENTION This invention provides a noviol olhs of tissue-selective ardrogen receptor Iating agents (ARTA). Ile agents define a new subclass of compounds which are S tissue-selootive androgen receptor moduleaors (SAWM, which are useful for oral IND testosterone replacement therpy, mate contraception, roaintaining sexual desire in women, osteopotosis, treating prostate cancer and imaging prostate cancer. These c-i agents have ani unexpected and tissue.-selective in-vivo activity for an androgenic and anabolic activity of a nonsteroidal ligand for the AI. These agents selectively act as 0010 Partial agoniSts in S0ome 1188008, while acting no fUll agonst in Other tiSSues, providing a a novel and unexpected meom for elloiting tissue-selective androgenic or anabolic effect. These agents may be active alone or in combination with progestins or estrogens. The invention fther provides a novel class of non-steridmi agonist compounds. The invewtion futher provides compositions containing the selective androgen modulator compounds or the non-stmtoidal agonist compounds and methods of binding an Alt, modulating sperrnatogenesis, bone formation and/or resorption, treating and imaging prostate cancer, and providing hormonal therapy for androgen-dependent conditions.

The present invention relates to a selective androgen receptor modulator compound having tissuo-selective in-vivo androgenic a=d anabolic activity of a rionsteroidlt ligand for the androgen receptor, the selective androgen receptor modulAtor compound repesented by the structure: of formula 1: 00 wherein X is a O. CH, NNSe, PR, or NR, Z is N0 2 CN, COP, COOH or CONHR; Y is L C 3 .Br, Cl, or Snlta; S Q is uiky halogen, N1a, NHCOCR 3 NHCOCF3, NHCOR, NHCONIR, NHCOOR, OCON R, CONR, NHCSCH 3 NMCSCI3, NHCSR NHSO2CH), 2 RK OR COR OCOR, 0S0 2 R, S02R or SR Ci wherein R is a aflyl, ay), hydroxy, C,-C 4 alkyl, a C,-C 4 baloalkyl, phonyl, halo, sikepyl or hydoxyl 010 or Qtoge iththbznne ringowhichitisattached isafusedring ByStem represented by structure A B or C: 0 0 NHO A C IS R 1 is CH3, CF 3 CU-CHs, or CF 2 CF,; and T is OHL OR, -NHCOC%, or NCOR wherein Ris a C 1

-C

4 alkyl, a CI-

C

4 haloalkyl, phenyl, halo, slknyl or hiydroxyl.

In one embodiment Q is in the par position. In aotber enbodiment X is 0.

In another embodiment, Q is in the pare.position and X is O. In yet another ernbodimeut4 Q is pam alkyl, halogen, NHCOCH 3 NHCOCI;3, NHCOR NHCONHRK NHCOOR, OCONHR, CONHR. NHCSCI, NHCSCF). NHCSR NHScH 3 NHSOA, OR, CO&. OCOk OSO2R, SOR or SR whorein R is a alkyl, aryl, hydroxy, Cl-C 4 uL8yl, Cl-C 4 haloakyl phenyl, halo, alkenyl or hyroxyl.

The present Invention relates to a selective androgen receptor modulator compound having !n-vivo androgenic and anabolic activity of a nonsteroldal Ligand for 00 the =&"mga rMqOWp, goe aelive mndroguu iecvpwr ModuWWo Dempatmd r~nemild by fth ano offImoala IL z Y NH Xj:r

H

3 C O wherein X is 0, CH 2 NH, Se, PR or NR; Z is a hydrogen bond acceptor, NO 2 CN, COR, CONHR; Y is a lipid soluble group, I,CF 3 Br, Cl, SnR 3 R is an alkyl or aryl group or OH; and Q is acetamido, trifluoracetamido, alkylamines, ether, alkyl, N-sulfonyl, 0- 0 sulfonyl, alkylsulfonyl, carbonyl or a ketone.

The prumt hvmdto so relous to a seilivo 4u&13gS mucptor moduilto E(IDpOmd baviung b,.vivo aodroguaic =n mabolic activiy df a ncmsturod figmn for UID mnd"im recepto velecve undm imcptor modulto ewupoad iupmeaWe by duo uktmtr of f(mmi ILh i mU X h a0. CH4 NI LSe. PR. ar NR Z B CN, OOL ar CONER, Y bu, CF 3 .Dr. C% ar SnRa; Sb OL oruy VA amor OxI amd Q bs scAtmdo or trulm0IMemnUlO.

730 Paumat AW~A= also to a seleatt"s audom ma epwr MOdult mnamd hWintuseldv ln-vtv =**Sonia and umbo& s advI of a 00 nonsteroidal ligand for the androgen receptor, the aeleotive androgen receptor modulator compound mproaentod by the structoe of formula IV:

CCF)

R

3

OH

00

IV

The present invention also relates to it selective androgen receptor modulator compound having tisaue-seicctive in-vivo androgeoic and anabolio activity of a rionstetoidal ligand for the androgen receptor, the selective androgen receptor modulator compound represented by the structore of fozmula V: 020 3p NH '0 OaOH

V

The present invention also relates to a selective androgen receptor modulator compound having tissue-selective in-vivo androgenic and anabolic activity of a nstCMoi c ligand fur the androgen receptor the selective androgen receptor modulator compound represented by the structure of formula V1 02N 3 NH, 0a

COC%

V1 00 Ct The present invention also relates to a selective androgen receptor modulator compound having tissue-selective iri-vvo androgenic and anabolic actiVity Of a c-i nonsteroidal ligard for the androgen receptor, the selective androgen receptor modulator comond represented by the structure of formoula VII: c-i02N NI[COCII 00 C3N NH 0H

VII

The present invention also relates to a method of binding a selective androgen receptor modulator comipound to an androgen receptor, which inchuda contacting the andirogen receptor with the selective mndrogen receptor modulator compound Under conditions effective to bind the selective androgen receptor modulator compound to the androgen receptor. In one embodiment the compound is Compound I. In another emnbodimnent the compound is Compound 11 In another embodiment the compound is Compound MI. Ini another embodiment the compound is Compound IV. In another wunboclirent the compound is Compound V. In 4nothe r exmbodiment the copud is Compound VI. In another embodiment the compound is Compound VII. In mnother embodiment the compound is Comipound VMI.

Another aspect of the present invention relates to 4 method of modulating spennatogenesis in a subject, which Includes contacting an androgen receptor of the subject with a selective androgen receptor modulator compound under conditions effecctive to increase or decrease spua prodction. In, one embodiment te compound is Compound 1. In another embodiment the compound is Compound IOL In another embodiment the compound is Compound JUL. in another embodiment te compound is Compound IV. In anotbar embodiment the compound is Comipound V. In ather 00 embodinmt the comnpoundi is Compound VI. In another embodiment the compound is Compound VII. In another embodiment the compound is Compound VMI.

The present invention also relates to a method of hormone therapy, comprising S contacting a androgen receptor of a subject with a selective androgen receptr mnodUlator compound under conditions effective to bind the selective androgen receptor modulator compound to the androgen receptor and effect a change in an androgendependent condition. In one embodiment the compound is Compound 1. In another embodiment the coMpouud is Compound 11. In another embodiment the comnpound is 00 to Compound MI. In another embodiment the compound is Compound IV. In another embodiment the compound is Compound V. In another embodiment the comlpound is Compound VJ. In another emnbodiment the compound is Compound VII. In another embodiment the compound is Compound VWI.

The present invention also relates to 4 method of treating a subject having a hormone related condition which comprises contacting an androgen receptor of said subject with a selective androgen receptor modulator compound under conditions effective to bind the selective androgen receptor modulator compound to the androgen receptor and effect a ohange in an androgen-clependent condition In one embodiment, the selective androgen receptor modulator compound is selective for androgen or testosterone receptor. The present invention also relates to a method of oral admin stration of the selective androgen receptor modulator compoun4 The present invention also relates to a method of tating a subject having a chronic muscle wasting disease slate which comprises otacting ani androgen receptor of said subject with a selective androgen receptor modulator compound as described herein under conditin.s offective to bind the selective aWrogert recetr modulator compound to the androgen receptor and effect a change in an ancirogern-dependent condition, In onie embodiment, the selective androgen receptor modulator compound Is selective for androgen or testosterone receptor. The present Invention also relates to a methodl of oral admuinistration of the selective andirogen receptor modualator compound.

In one embodiment the compound is Compound L. In another embodiment the 00 0 0 compound is Compound 1. In another embodiment the compound is Compound 1. In c- another embodiment the compound is Compound IV. In another embodiment the Scompound is Compound V. In another embodiment the compound is Compound VI. In Sanother embodiment the compound is Compound VII. In another embodiment the compound is Compound Viii.

CAs defined herein, the disease state of "chronic muscle wasting" means 00 The present invention also relates to a method of treating a subject having 0 prostate cancer which comprises administering to a subject an effective amount of a C1 selective androgen receptor modulator compound. In one embodiment, the selective androgen receptor modulator compound is selective for androgen or testosterone receptor. In one embodiment the compound is Compound I. In another embodiment the compound is Compound II. In another embodiment the compound is Compound I. In another embodiment the oompomund is Compound IV. In another embodiment the compound is Compound V. In another embodiment the compound is Compound VI. In another embodiment the compound is Compound VII. In another embodiment the compound is Compound VIIL.

The present invention also relates to compositions and a pharmaceutical compositions which comprises a selective androgen receptor modulator alone or in combination with a progestin or estrogen and a suitable carrier, diluent or salt In one embodiment the composition comprises Compound I. In another embodiment the compound is Compound IL In another embodiment the compound is Compound HI. In another embodiment the compound is Compound IV. In another embodiment the compound is Compound V. In another embodiment the compound is Compound VI. In another embodiment the compound is Compound VI. In another embodiment the compound is Compound VIII.

The present invention relates to a non-staroidal agonist compound, the nonsteroidal agonlst compound represented by the suucture of formula VIII: 0 "T o0 Vm 00 Wherein X is a0.CH 2 ,NIL.So, PR, orNR; R, is Cli,, CK 3

CH

2

CH

3 or CF 2

CP,;

T is OH, OR, -NHCOCH 3 or NRCOR whereini R is a C,-C 4 alkyl, a C 1

C

4 blaloalkyl. PhonyI. halo, alkwyI or hydroxyl: A is a 5 or 6 membered saturted, unsaturated or aromatic carbocyclic or heterocyclic ring iresonmted by the stnitewre: Z 2 A A- or Y"<1

A

7 1is a 5 or 6 imombore4 6aXtr~ted, msatrated or aromatic carbocyclic or heterocyclic ring represented by the stm~oture:

B

4 or Q2rY W;

B-

wherein Av At 1 are each C,0, S rN; 13-Btare each C,0, S orN; Z iS NO 2 CN, COOH COP, or CONHR, Y is 1, CP3,,Sr. C1. or SnR3; and Q I and 02 ame independenty of each other alkyl, halogen. NR 2 NHCOCH3. NHCOCF,, NHCOR, NHCONHR, NHCOOR, OCONHR. CONHR.

NIICSCfl,, NUCSCFi, NHCSPR NHS02CH3. NHSOA~ ORt. COR. OCOR.

OSOAZ S0 2 R or SR wherein R is a CI-C 4 alkyI, a C,-C.4 haloalkyl, phanyl, halo.

alkeyl or hydroxyl.

00 The Present invention also relates to a com1positon and pharnnaeutical comnposition comprising the non-steroidal agonist compound alone or in combinationi with a Progestin or estrogen and a suitable cardler, diluent or salt. In one embodiment the compound is Compound 1. In another embodiment the compound is Compound 11. In another embodiment the compound is Compound 1U. In another embodiment the compound is Compound IV. In asnothier embodiment the compound is Compound V. In another embodiment the compound is Compound VT. In another embodiment the compound is Compound V[U. In another cinbodinent the compound is Compound VQI.

00 t ThU present invention also relates to a method of binding a non-steroidlal agonis compound to 4n androgen receptor comprising contacting the androgen =eeptor with the non-steroidaI agonist compound wuder conditions eftective to bind the nonsteroidal agonist compound to the androgen receptor. Ini one embodiment the compound is Compound 1. In another embodiment the compound is Compound H. In another Is embodiment the compound is Compound MI. In another ernbodiment the compound is Compound IV. In another embodiment the compound is Compound V. In another embodiment the comnpound is Compound V1. In anotber embodiment the compound is Compound V11. In another embodiment the compound is Compound VIMh The present invention also relte to a methad of modulating spermtogenesis in 4 subject comprising contacting an androgen receptor of the subject with a nonsteroidal agonist compound under conditions effective to increase or decrease sperm Production. In one embodiment the compound is Compound 1. In another ernbodimner the compound is Compound 11. In another embodiment the compound is Compouznd Ml In another embodiment the compound is Corupomid IV. In anothat embodiment the compound is Compound V. In another embodiment the compound is Compound VI. In another embodiment the compovnd is Compound VT. In another ombodivicat the compound is Compound VII.

The present Invention also relate to a method of hormone therapy comprising contacting an miadrgeu "eetor of a subject with a uort-atmidal agonist under conditions effaoto to bind the nomoeeroWaI agonist compound to the mndrogen receptor 00 and effect a change in an androgen-depondent condition. in one embodiment the compound is Compound L In another emibodliment the compound is Compound 11. In another embodiment the compound is Compound MT. I aniother embodiment the- C~1 compound is Compound IV. In another embodiment the compound is Compound V. In S nother. embodiment the compound is Compound VI. I another embodiment the compound is Compound VT!. In another embodiment the compound is Compound VMI.

The present invention also relates to a method of treating a subject having a 00 hormone related condition which comprises contacting an androgen receptor of said subjeot with a non-steroidal agonist compound under conditions effective to bind the non-steroida) agonist compound to the 4ndrogeui receptor and eftet a change in an androgen-dependerit condition. In one embodiment. tho non-steroidal agonist compound is selective for androgen or testosterone receptor. The present invention also relates to a method of oral administration of the non-steroidal agonist compound. Ir one embodiment the compound is Compound I. In another emubodimont the comipound is Compound 11. In another embodiment the compound is Compound MI. In another embodiment the compound is Compound IV. In anodhe embodiment the ompound Is Compound V. in another embodiment the compound is Compound VL In another embodiment the compound is Compound V1I. In another embodiment the compound is Compound VUI The present invention also relates to a method of treating a subject having prostate cancer which comprises administrating to a subject an effective amount of a non-steroidal agonist compound. In one embodiment, the non-steroida) 4gonist compound Is selective for androgen or testosterone receptor, In one embodiment the compound Is Compound 1. In another embodiment the compound is Compound II. In another embodiment the compound is Compound MT. In another embodinmt the compound is Compound TV. In another enibodinnent the compound is Compound V. In another embodiment the compound is Compound VI. i another embodiment the compound is Compound V11. In another embodiment the compound is Compound VMI.

Still another aspect of the present relates to a method of producing a selectve 00 androgem receptor modulator or a non- steroidal AR agonist com~pound of the present invention. In one anibodliment the compound is Compound 1. In another embodiment the compound is Compound 11, In another embodiment the compound is Compound TU. In C~~1 anoth=em ibodimnent the compound is Compound IV. In another embodiment the compound is Compound V. In another embodiment the compound is Compound VI_ In another embodiment the compound is Compound VIT. In another embodiment the compound is Compound Vli.

0 0 10 of a selective androgen modulator compound and/or a non-sternidal agonist compound of the present invention in a sample. The method comprises the steps of obtaining the sample, and detecting the compound in the sample, thereby determining the presenoe of the compolmd in the sample. In one embodiment the sample is a blood serum, plasma, urine, or saliva, sample. la another embodiment the detection step comprises measurng 1S the absorbance of the compound. In one embodiment the compound is Compound IL In another embodiment the compound is Compound IL I another embodiment the compound is Compound Ill. In another embodiment the compound is Compound TV. In =nother embodiment the compound is Compound V. In nother embodiment the compound is Compound VI. In another embodiment the compound is Compound VI1. In another embodiment the compound is Compound VMi.

The novel selective androgen receptor modulator compounds and the nonsteroidal agonist compounds of the present inveation, either alone or as 4 oomposition, are WOeW in malies and females for the treatment of a variety of hormone-related conditions, such as hypogonadism, sarooponla, oythropoicsis, erectile function Wak of libido, osteoporesis and fertility. Further, the selective androgen receptor modulator compounds and thea non- steroidal agonist compounds are usefnl for oral testosterone replacement therapy, treating prostate cancer, imaging prostate cancer, and maintaing sexual desire in~ women. The agents may be used alone or in combination. with a progestin or estrogen. In one embodiment the compound is Compound L. In another embodiment the compound is Compoundi T1. in another embodiment the compound is Compound THI. In another embodiment the compound Is Compound TV. In another 00 embodiment the compound is compound V. In another embodiment the compound is Compound VI. In another embodiment the compound is Compound VII. rxn another embodiment the compound is Compound VIII S The selective androgen receptor modulator compounds and the non-steroidal agonist compounds of the present invention offer a significant advance over steroidsl androgen treatment because the selective androgen receptor modulator compounds and the non-sterida! agonist compounds of the present invention have been shown in-vivo 00 to have a tissue-selective androgenic and anabolic activity of a nonsteroidad ligand for the androgen receptor. Moreover, the selective androgen receptor modulator compounds and the noni-steroidal agonist compounds of the present invention are not accompanied by serious side effects. latbility to oxidative metabolism, inconvenient modes of aduinistrafion, or high costs and still have the advantages of oral bioavailability, lack of crss-reactivity with other steroid receptors, and long biological half-lives. In one 1S embodiment the compound is Compound I. In another embodiment the compound is Compound 11. In another embodiment the compound is Compound MI. In another embodiment the compound is Compound TV, In another embodimemt the compound is Compound V. In another embodiment the compound is Compound VI. In another embodiment the compound is Compound V1I. In another embodiment the compound is Compound VII.

BRIEF DES CRIPTION OF THE DRAWINGS The present invention will be understood and appreciated more fIlly from the following detailed description tW=e in conjunction with the appended drawings in which.

Figure 1. Androgenic and Anabolic activity of (S)-GI'x-007 in rats, Rats were left untrested (intact control), castrated (castrated control), treated with testosterone propionate or treated with S-07 x-007, and the body weight gain as well as the weight of androgen-responsive timses (prostate, samirnal vesicles and lavator an! musole) vas determined.

00 Figu~re 2; Androgenic and Anabofic activity of S-OTxh,007 in rats. Rats we=e left untheated (intact control), castrated (castrated control), treated with 0.1, 0.3, 0.5,9 0.75 and 1.0 mg/day testosterone propionate or treated with 0. 1, 0.3, 0.5, 0.75 and 1.0 mg/day SOTx-007, and the weight of androgen-rcsponsive IND tissues (prostate, semimal vesicles and levator ani muscle) was determined.

Figure 3: Androgenic and Anabolic activity of S-GTx-014 inl rats. Rats were left 00 untreated (intact control), castrated (castrated control), treated with 0. 1. 0.3, 0.5, 0.75 and 1.0 mg/day testosterone propionate or treated with 0. 1, 0.3.

050.75 and 1.0 mg/day S-G1'x-014, and the weight of androgen-respousive tissues (prostate, semimal vesicles and levator an rmuscl~e) was determined.

Figure 4: Avenage plasma ooncentration-time profiles of S-GTx-007 in beagle dogs after IV administration at 3 and 10 mg/kg.

Figure Average plasma concentration-time profiles of S-GTx-007 in beagle dogs after PO administration as solution at 10 mg/kg.

Fig 6: Averge plasmae conceatratlon-tinie profiles of S-GTx-007 in beagle dogs after IV admifistration as oapsules at mg/kg.

Figure 7: Uffects of 01"x-014 and GTx-007 on LII Levels.

Figure 8: Biects of GTx-014 and GTx-007 on FSH Levels.

Figure 0: Synthesis sch~eme of OTX-007- 00 DETAILED DESCRIPTION OF THE INVENTION This invention provides a novel clas of androgen receptor targeting agents CI (ARTA). The agents define a now subclass of compounds which are tissue-selective Sandrogen receptor modulators (SAR.M) which axe useful for oral testosteroue INOreplacement therapy, male ourraception, maintaining sexul desire in women, treating prostate cancer and imaging prostate cancer. These agents have 4n unexpected tissueselective in-vivo activity for aa androgenic and anabolic activity of a nonstercidal ligand 00 for the androgen receptor. These agents may be active alone or in combination with progestins or estrogens. The invention finther provides a novel class of non-steroidal agonist compounds, The invention fizrtber provides compositions containing the selective androgen modulAtor compounds or the non-steroidal agonist compounds ad methods of binding an andrdgen receptor, miodulating spennatogenosis, treating and imaging prostate cancer, and providing bormonal therapy for androgen-dependeifl conditions.

The corupunds described herein, define a new class of selective androgen receptor modulators (SARMS) that demonstrate potent anabolic effects muscle growth) with less androgenic activity prostatic growth). This new class of drugs hasSeveral advantages over non-selective androgens, including potential therapeutic applications in males and females for modulation of fertility, eiytliropoiesis, osteoporosis, sexual )ibido atid in men with or at high risk for prostate cancer. In one embodiment the compound is Compound 1. In another embodiment the compound is Compound 11. In another embodiment the compound is Compound Ml. In another embodiment the compound is Compound IV. In another embodiment the comnpound is Compound V. In another embodiment the compound is Compound VI. In another embodiment the compound Is Compound VII. In another embodiment the compound is Compound VMI Further, in one embodiment the compunds have tissue specific pharmacologic activity. As demonstarted in Figure 7 and 8, GTx-007 does not suppress LII levels at doses that ate capable of eliciting maximal stimulation of levator ani muscle growth and 00 does not suppress ESH levels at doses that are capable of eliciting maximal stimulation of levator ani muscle growth.

The present invention relates to a selective androgen receptor modulator s compound having tissue-selective in-vivo androgenic and anabolic activity of a nonsteroidal ligand for the androgen receptor selective androgen receptor modulator compound represented by the structure of formula 1: 00 00

Q

I R

T

I

wherein X is a 0, CH2, NH, Se, PR, or NR; Z is NO 2 CN, COR, COOH or CONHR; 1o Y is 1, CE 3 Br, Cl. or SnR 3 Q is alkyl, halogen, NR2, NHCOCH 3

NHCOCF

3 NHCOR, NHCONHR, NHCOOR, OCONHR, CONHRK NHCSCHJ, NHCSCEI, NHCSR NHS0 2

CH

3 NHSO2R. OR, COR, OCOR, OSO 2 R, SQ 2 R or SR wherein R is an aryl, Cr04 alkyl, a C 1

-C

4 haoalkyl, phenyl, halo, alkenyl or hydroxyl; or Q together with is the benzene ring to which it is attached is a fused ring system represented by structure A, B or C: O 00 A C 00 It is Ciis, Ch, CH2CH). ot CF 2 C~i; and T is OH. OR,. -N1HCOCJ%, or NUCOR Whma R is a CI-C hikl, a Ci- CKI C hal"W.yl phenyL halo. aikanyl orhydmxyt.

In am ecmbodiment. Q is in the pama positio of thei beiza rng to which i Is AU113-hed u&b embodiet Qtisinepaipoiion an4 X isO. Inm=other emboditneat, Q is in the pan position and is .41yL, halogen NR2, NHCOCH3, 00 NHCOCF3. NUCOR. NHOHR, NIHCOOR, OCONHR. CCYNHR NUCSCH3.

NHCSMZ

3 NJHCSR NHSO 3

CH

3 W QA~R OP, CO&l OCOXt OSOAR SO3R or SR Wheavin R is a, uzyl, C 1

-C

4 AAyt a CI-Cs baOaELkl pbanyl, Wao. alkwzyt or hydnrxyI.

The posait invention utlat o a selctvo androgen receptor inodulato Is S compound having i-vivo adrogeic and uzzabolic actt of at nanaexcidal lgad for the aftdro septor. the seleclive androigen receptor nodiilawo compound repreented by the strutr of formula 1: Y NHX H1 3 C "OH wherein X is 0, CH 2 NH, Se, PR or NR; Z is a hydrogen bond acceptor, NO 2 CN, COR, CONHR; Y is a lipid soluble group, I,CF 3 Br, Cl, SnR 3 R is an alkyl or aryl group or OH; and Q is acetamido, trifluoracetamido, alkylamines, ether, alkyl, N-sulfonyl, 0sulf'onyl, alkylsulfonyl, carbonyl or a ketone.

00 The Present invention also relates to a selective androgen receptor modulator compound baving in-vvo acrogenic and anabolic activity of a nonstoroldal ligarid for the androgen receptor tho, selective androgen receptor modulator compound represented c-i by the strucure of formula EII: Ci Q 00 s where X is a,CH 2 NH, So, P&orNR; Z is N02 CM, COR, -or CON*IR; Y is r. CF 3 Br, CI, or SuR3; R is analkyl or arylgroup or OH; and Q is acetarnido or trifluracetamido.

The present invention also rMates to a selective androgen receptor modulator compound having tissue-selective in-vivo androgenic and anabolic activity of a nonsteroidal ligand for tbe androgen receptor, the selective androgen receptor Modulator compound represented by the structure of formula IV: 0 2

N

The present invention also relates to 4 selective androgen receptor modulator compound baving tissue-selective in-vivo androgenic and anabolio activity of a nonsteroidal ligand lbr the androgen receptor, the seletive androgen receptor modulator compound represented by the structure of fon-aulat V: 00 00 The present invention also relates to a selective androgen receptor modulator compound having tissue-selective in-vivo androgenic and anabolic activity of a S nonstotoidal ligand for the androgen receptor, the selective androgen receptor modulator compound represented by the structure of formula VI: 02N COCH 2 CHt 3 0 2 0 CF3 3 OHK j

VI

The present invention also relates to a selective androgen receptor modulator compound having tissue-selactive in-vivo androgenic and anabolic activity of a nonsteroidal figand for the androgen receptor, the selective androgen receptor modulator compound represented by the structur of formula VII: The present invention relates to a non-steroldal agonist compound having the is formula (Compound VIII): 00

NH

vm s wherein X is a 0, CH 2 NH, Se. PXor NR; R, is CH 3 CF3, CH 2

CH

3 or CF 2

CF

3 T is OIL OR, -NHCOCH3, or N1COR wherein wherein R is a C 1

-C

4 alkyl, a C 1 -C haloalkyl. phenyL hao. aUnyl or hydroxyl; A is a 5 or 6 memberod s9tated, unsatated or aromatic curbocyclic or 0 heterocyclic ring represented by the swuoture: Z A3----A 2 A; Al- or x9 -A AA7- A16 B is a 5 or 6 membered satuamed, unsaurated or aromatic carbacyclic or heterocyclic ring represented by the shuoture: 14 or Be~ Qv 7- QZ Bf 00 wherein AI- are each C, 0. S or N; Be- BII ameach C 0. S or N; IN Zis No], CN, COk~ COOK, or C0NHRl; Y'S1, CiF 3 BrCl, Or SnJ3; anid Q, and Q2 arm independently of each other alkyl, halogen, NR2, NWiCOCH, NHiCOCF 3 NHCOP, NHC0NI; NHCOOROOpRCNR 00 NHCSCfj,,

NHCSCF

3 NHCSR NHSo 2 CH3, NHS0 2 R, O& C01t, OCOR, 0S0 2 R, S02R Or SR~ wherein R is a Ci-C 4 alkyl. a CI-C4 haloalkyl, phonyl, halo, alkenyl or hydroxyl.

The substintents Z and Y can be in any Position ot the five or 6 mnembered ring carrying these substtutents (hereinafter A ring"). SiMilarlY, the Sbstituent Q an be in 'S any Position of the five or 6 memnbered ring canying this substitutent (hereinafter

"B

ring"). It is unlderstood that when any of the ring memibers A,,I or

B

11 are 0 or S, then these ring members axe unsubstituted. It is fuirther underttood that when any of the ring members All or 13 1 Bli are 0 or S, then the dotted line between said ring MOen and other ring members represents a single bond.

In one ernbodiinent, the A ring includes any tye of saturated or unsaturated oarbocyclic ring. In one embodiment, the A ring is a 6 membered saturated carbocyclic ring, which may be unsubstituted. inonosubstittd or polysubstitute4 by any of the substitutents described hereinabove. In one embodiment, the A ring is a 5 memabered satuated cabocyclic zing, which may be unsubstituted, ruonosubstituted or polysubstituted by any of the substitutents described hereinabove. In another em bodiment, the A ring is a 6 nmmbered carbocyclic ring containing one or more double bonds, which ring may be untsubstituted, monosubsltte4 or polysubstitated by any of the substitutents described hereinabove. In another emnbodiment the A ring is a nrimbered carbocyclic ring containing one or more double bonds, which ring may be Unsubstitt, monosubstitue or polysUbstitu.ted by any of the substitutents described hereinabrjve.

00 [f00049) In another embodirrent the A ring includes any type of saturated, unsaturated or aromatic heteocyclic ring. In another embodim~ent, the A ring is a 6 membmad CIaturated heterocyclic ring, which may be unsubstituted, nonosubstituted or polysubstituteci by any of the substituents described hereinabove. In another IND embodiment, the A ring is 4 5 membered saturated haterocyclirc ring, which may be umaubstituted, monosubstituted or polysubstituted by any of the gubstitiients described hereinabove. In another embodiment, the A ring is a 6 membered heterocyclic ring 00ontaining one or more double bonds, which ring may be unsubstituted, monosubstituted or polysubstituted by any of the substitutents described hereinabove. In another embodiment, te A ring is a 5 membered heterocyclic ring containing one or more double bonds, which ring may be unsubstituted monosubstituted or polysubstituted by any of the substitutents described hereinabove. In another embodiment the A ring is a 6 memnbered hoteroaromatic ring which may be unsubsttuted, monoaubstitut-ec or polysubstituted by any of the substitutants described hereinabove. In another embodiment the A ring is a 5 memnbered heteroarornatic ring which may be unsubstituted, monosubstitutod or polysubstituted by ay of the substitutents described hereiabove.

Similarly, the B ring includes any "ye of saturated or unsatred carbocyclic ring. In one embodiment, the B rig is 4 6 mrembered saturate earbocyolic ring, which may be unisubstituted, ruonosubstituted or polysubsttted by any of the substitutents described hereinabove. In one embodiment, the B ring is 4 5 membered saturated carbocyclic ring, which may be unsubstitutod. monosubstitutod or polysubstituted by any of the substitutents described hereinabove. in =nother wmbodiment, the B ring Is a 6 memubered carbocyclio ring containing one or imore double bonds, which ring may be unsubstituted, rnono'substitute or polysubsttUrtod by any of the substitutents described hereinabove. In another embodiment the B ring is a 5 memaberedi carbocyclic ring containing one or more double bonds, which ring may be unsubstituted, rnonosubatitutecl or polyaubstituted by any of the substitatents deacribed hereinabove.

00 In another embodiment the B ring ficcles any type of saturated, Unsaturated Or aromatic heterocyclic ring. In another embodiment, the B ring is a 6 mnembered sawurted hieterocyclic ring, which may be unsubstituted, inonosubstituted or CI polysubstituted by any of the substituents described hereiabove. In another einbodimn~ the B ring is a 5 memnbered saturated t~etrocyclio ring, which may be INDunsubatintUd, monosubstitated or polysubstiuited by any of the substituons described hereinabove. In another embodiznent, the B ring is a 6 memnbered baterooyclic ring containing one or more double bonds, which ring may be unsubstituted. inonoaiubstituted 00 or polysuibstituted by any of the substitutents described herainabove. In another to embodiment, the B ring is a 5 mnembered heterocyclic ring containing one or more double bonds, which ring may be inubstizuted monosubstituted or polysubstitaled by any of the substitutets described hereinabove. In another embodiment, the B ring is a 6 rnezubered beteroaroxnatc zing which may be unsubatituted, inonosubstitle4 or polysubstituted by any of the substtutents described hereinabove. In another 1S embodiment the B3 ring is a 5 mnembered heteroaromatic ring which may be unaibstitutrd, inanosubstituted or polysubstituted by any of the substitutents described hereinabove.

Nonliraiting examples of suitable A rings and/or B rings are carbocyclic, rings such as cYrc)opeftafle, cyclopentene, cycloheicane, and cyclohexene rings. and heteiocyclie rings such as pyran, dihydropyran, tetrahydropyran, pyrnole, dihydropyrrale, tethyciropyrrole, pyrazine. dihydropyrazine, tetrabydropyrazine, pyriinidine, diydropyrimidine, temraydropyriinidone, pyrazoL, dihydropyrazol. tetr-ahydropyrazol, piperidine, piperazine, pyxidine, dihyctropyridine, tetrahydropyridine, morpholine, thlornorpholine, ftran, dihydrolbran, tshydrofuran, thiphene, dihychuthiophene, teftahydrothlophane, Whaole, imidazole, isoicazole, and the like.

As used herein, receptors for extranllula signaling molecules ac collectively referred to as "cell signaling receptors". Many a signaling receptors are tmnsmemnbrane proteins on a cell ourface; when they bind an extraceihilar signaling molecule a ligan4l they become activaed so as to generate a cascade of intracellular signals that alter the behavior of the cell. In contrast, in some eases. the 00 receptors are inside the call and the signaling ligand has to enter the cell to activate then; those signaling molecules therefore must 6a sufficiently smnall and hydrophobic to diftws across te plasma membzane of the cell. As used heroin, these receptors arm CIcollectively referred to as "intraoalulrJ call signaling receptors".

Steroid hormones are one example of smoall hydrophobic molecules that diffgse directly across or ane transported across the plasma membrane of target cells and bind to intracellular call signaling receptors. These receptors ame structuialy related and 00 constitute the intracellular receptor superfamily (or steroid-hormone receptor supesfamily). Steroid hormone recaptors include progefterono receptors, estrogen receptors, androgen receptors, gbroocorticoid receptors. and uiinerslooorticoid, and flumserU orphan receptors. The present invention is particutarly directed to androgen receptors and all of its isoforrns.

is In addition to ligarid binding to thte receptors, the receptors can be blocked to prevent ligand binding. When a substance binds to a receptor, the three-dimensional structure of the substance fits into a space created by the thrree-dimensional stracture of the receptor in a ball and socke ooniflguration.

The better the ball fits into the sockcet, the more tightly it is held. This phertorenou is called affinity. If the affinity of a substance is sufficiently high it will compete with the hormone and bind the binding site more frequently. The binding of the liganci may also lead to tissue-selectilve recruitment of other important proteins to trarsduce the signal. These proteins are known as coactivators and oopressor.

participtte in signal transducdion, and way be selectively induced or inhibited by Uigand binding. Once bountds signals may be sent through the receptor into the cells, causing the call to respond in some fashion.. This is caled activation. On activation, thle activated receptor then directly regulates the trascription ofspecific genes. Rat the substance and the receptor may have certain attributes, other than aftity. toa activate the call.

Chemical bonds between atoms of the substance and the at*=s of the receptors may form. In some cases, this leads to a change in the configuration of the receptor, which is enough to begin the activation promes (caled signal mnusduction). As a result, 00 substances can be made which bind receptors and activate them (called receptor Ct 4gonists) or inactivate them (called receptor antagonists).

N- The present invention is directed to selective androgen receptor modulaor compounds which are agonist compounds, and are, therefore, useful in binding to and IND activating steoidal hormone receptors. The compounds are non-steroidal. Preferably, the c-i agonist compound of the present invention is an agonist that binds the androgen receptor. Praferably, the compound tos high affinity for the androgen receptor. The 00 compound may bind either reversibly or izreveribly to the androgen receptor. The cornpolnd of the present invention may contain a fUnctonal group (afinity label) that allows alkylation of the androgen receptor covalent bond formation). Thus, in this case, t compound binds irreversibly to tho receptor and, accordingly, cannot be displaced by a steroid. such as the endlogenous ligands dihydrotestosterone and testosterone. It is preferable, however, for the compounds of the present invention to reversibly bind the androgen receptor.

According to one aspect of the present invention, a method is provided for binding the selective androgen receptor modulator compounds of the present inventon to an androgen receptor by contacting the receptor with a selective androgen receptor modulator compound inder conditions efetive to cause the selective 4ndrogen receptor modulator compound to bind the. andrkogen receptor. The binding of the selective androgen receptor modulator compounds to the androgen receptoi enables the compounds of the present inventiou to be uasefu~l in mates and in females in a number of honmone therapies. The agonist ompounds bind to and activate the androgen receptor.

Binding of the agonist compound is either reversible or irreversible, preferably reversible.

According to one aspect of the preset invention. a method is provided for modulating spermatogenesis by contating an mndrogen receptor of a patient with a selective androgen receptor modulator compound under conditions effective to bind the selective androgen receptor modulator compound to the androgen receptor and incre or decrease sperm production.

00 According to another aspect of the preent invention, a method is provided for hormnal therapy in a patient suffering from an androgen- dependent condition) C1 wbich includes contacting an androgen receptor Of a patient with a selective androgen receptor modulator compound tunder conditions effective to bind the selective androgen IND receptor modulator compound to the androgen receptor and affect a ohange in an androgon-depondent condition. Azdrogen-dependont conditions that may be treated according to the present invention include those conditions associated with aging. such 00 as hypogonadisn, sarcopea, erytbropoiesis, osteoporosis, and any oter conditions later determined to be dependent upon low androgen testoistarone) levels. In one embodiment the selecve androgen receptor modulator compound is administered alone. In another embodiment, the selective androgen receptor modulator compound is administered in combination with progestin. In yet another embodiment, the selective androgen receptor modulator compound is administered in combination with estrogen.

According to another aspect of the present invention, a method is provided for treating a subject having prostate cancer. The method comprises administrating to a subject an effective arnount, of a selective androgen receptor modulator compound. In one embodiment, the selective androgen receptor modulator compound is selective for androguen or testosterone receptor.

The present invention also relates to a method of treating a subject having a chronic nmscle wasting disease state which comprises contag ant androgen receptor of said subject with a selective androgen receptor mnodulator compound under conditions effective to bind the selective andirogen receptor modulator compound to the aidrogen receptor and effect a change in an androgen-clepondent condition. In one embodiment, the selective mndrogen receptor modulator compound is selective for androgen or testosteron, receptor. The present invention also relates to 4 method of oral administration of the selective androgen receptor modulator OOMPOUnd. In Oo embodiment the comupound is Cpound i. in another embodiment the compound is Compound 11. In another embodiment the compound is Compound IH. In another embodiment the compound is Compound IV. in another embodiment tlw compound is 00 Compouind V. In another embodiment the compound is Compound V1. In another embodiment the compound is Compound VII. In another embodiment the compound is Compound VII.

According to one aspect of the present invention, a method is provided for binding the non-steroidal agonist compounds of the present Invention to an androgen receptor by contacting the receptor with a non-steroidal agonist compound under 00 conditions effective to cause t non-steroidal agonist compound to bind the androgen receptor. The binding of the non-steroidali agonist compounds to the androgen receptor enables the comipounds of the present invention to be usefol in roales and in females in a number of hormone therapies. The agonist compounds bind to and activate the androgen receptor. Binding of the agonist compound is either reversible or irreversible, preferably reversible.

According to one aspect of the present invention, 4 method is provided for modnating spennatogenesis by contacting an andirogen receptor of a patient with a nonsteroidal agonist compound under conditions effective to bind the selective androgen receptor modulator compound to the androgen receptor and increase or decrease sperm production.

According to another aspeot of the preet invention, a method is provided for hormonal therapy in a patient one suffering from an adrogen- dependent condition) wbiob Includes contacting an androgen receptor of a patient with a nonsteroidal agonist oompownd under conditions effective to bind the non-steroidal, agonist compound to the 4ndrogen receptor and effet at change in an Androgen-depandent condition. Androgen-dpendent conditions thet maSy be treated aczoording to the present invention include those conditions associated with aging, such as hypogonadism sarcopenia, erythropoiesis. osteoporosis, lack of sexua libido and any other conditions later determined to be dependent upon low androgena testosterone) levels. In one embodiment the non-s tmridal agonist compound is administered alone. In another embodiment, theo non-steroidal agonist compound is administered In combination with 00 progestin. In yet another embodiment, the non-steroidal agordst compound is administered iii combination with estrogen.

According to another aspect of the present invention, a method is provided for S treating a subject having prostate cancer. The metod comprises administrating to a IND subject an effective amount of a non-steroidal agonist compound. In one embodiment, (71 the non-steroidal agonist compound is selective for androgen or testosterone receptor.

00 ~The compounds of the present invention have an assymetrc center and can be the FR or S isomer, or a mixtue of both. In one embodiment, the compounds raceinic mixtures of the Rt and S enantiomers. In another embodiment, the comnPounds are substantially pure Rt enantiomers. In another embodiment. the compounds are substantially pure S enantioniors. "Substantialy pure" is defined herein as greater than about 95%h preponderance of one Isomer. Whore the above-described processes for the preparation of the compounds of use in the invention give rise to mixtures of stereoisomers, these isomers may be separated by conventional techniques, such as preparative chromatography. The compounds may be prepared in racetuic form, or individual enantiomners may be prepared either by enantiospecific synthesis or by resolution As used herein, "phamlawcutical composition" means therapeutically effective amiounts of the MAM or the non-steroidal agonist compound of the present invention, together wit suitable Qiuents. preservatives, solubilizers. emuluifiers, adjuvant and/or carriers. A "therapeutically effective amount" 4asused herein refers to that Amount which provides a therapeutic effect for a given condition and administration regimen, Such omnpositions are liquids or Lyophilized or othewise dried fortnulations and Include diluents of various buffer contentl Tris-HCL, acetate, phosphate), pH =nd ionic stregth, additives such as albumin or gelatin to prevent absorption to surfaces detergents Tweem 20. Tween 80, Pluronic P68. bile atid salts), solubilizing agents glycerol, polyetylene glycerol), anti-oxidants (egasorbiic acid, sodium metabisullite), preservatives Thimerosal, benzyl alcohol, pmrbens), bulking substances or tonicity modifiers lactose, moaunitol), covalent attachment of 00 polymers such as polyethylene glycol to the protein, complexation with metal ions, or Incorporation of the material into or onto particulate prparations of polymeric compounds such as polylactic acid, poiglycolic acid, bydrogels, etc, or onto liposomes, microoulsions, micalles, uvilameUar or multilameilar vesicles, erythrooyte ghosts, or s spheroplasts. Such compositions will influence the physical'state, solubility, stability, rate of in vivo release, and rate of in vivo clearance. Controlled or sustained release compositions include formulation in lIpophilic depots fatty acids, waxes, oils).

Also comprehended by the invention are particulate compositions coated with polymers poloxamers or poloxamines). Other embodiments of the compositions of the invention incorporate particulate forms, protective coatings, protease inhibitors or permeation enhanoers for various routes of administration, including parentoil, pulmonary, nasal and oral. In one embodiment the pharmaceutical composition is adminiterd parenterally, paracanceratly, trunamucosally, trqnsdermally, intramuscularly, intravenously, Intradermally, subcotaneously, intraperitonealy, intraventrioularly, intracranially and intratumorally.

Further, as used herein "pharmaceutically acceptable carriers" am well known to those skilled in the art and include, but ae not limited to, 0.01-0.IM and prefbrably O.05M phosphate buffer or 0.8% saline. Additionally, such pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions.

Examples of non-aqueous solvents ae propylene glycol, polyethylene glycol. veigetable oils snob as olive oil, and injectable organic esters such as ethyl oleate. Aqueous caniers include water, alcoholic/aqueous solutions, emulsions or suspensions. including saline and buffered media.

Parentoral vehicles include sodi=m chloride solution. Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils. Intavenous vehicles include fluid and nutrient replenishers, electrolyte repleniahers such as those based on Ringer's doxtrose, and the like. Preservatives and other additives may also be present, sqch as, for example, antimicrobials, antioxidants, collating agents, inert gases and the like 00 Controlled or sustained release compositions include formulation in lipopbilir.

depots fatty aoids, waxes, oils). Also comprehended by the invention are pardiculate c-i compositions coate with polymers poloxame r poloxamines) and the compound coupled to antibodies directed against tissue-specific receptors, ligands or anti gens or IND coupled to ligands of tissue-spcifc receptors.

Other embodiments of the compositions of the invention incorporate 00 paticlate forms, protective coatings, protease inhibitors or permeation enhancers for various routes of administration, including parenteral, pulmonary, nasal and oral.

Comnpounds modified by the covalent auacbrnent of water-soluble polymaer such as Polyethylene glycol. copolymers of polyethylene glycol anid polypropylene glycol, carboxyrnorhyl cellulose, detn, polyvinyl alcohol, poiyvinylpyrrolidotie or polyproline ame known to exhibit substantially longer half- lives in blood following intravenous injection than do thne corresponding umnodified compounds (Abuchowski ct al., 1981; Newmnark et a&L, 1982; and Kaftr et al., 1987). Such modifications may also increase the compound'~s solubility in ueous solution, elimninate iggregation, enihanc the physical and chemical stability of the compound, and greatly reduce the imxrunogeniy anid reactivity of the compound. As a result, the desired in vivo biological activity my be achieved by the administration of such polymer-compound abducts less frequently or in lower doses than with the unmodified compound.

In yet another embodimnt, the pbarmaceutical composition can be delivered in a controlled release system For oxmple, the agent may be administered using intravenous infusion, an implantable osmotic pump, a tuusdermal patch, liposornes, or other modes of administration. In one embodiment, a pump may be used (see Langer.

supra; Sefton, C1~.C Crit. R&e Biomad Rag. 14:201 (1987); Buohwald at al., Surgery 88:507 (1980); Saudelc ot al., N. BaZIg. 1. Med. 321.574 (1989). bn another embodiment, Polymeric Materials Can be used. In yet another embodiment. a controlled release system can be placed in proximity to the thieautio target, the brain. thus requiring only a fraction of the systemic dose (see, (3oodson, in Medical Applications of Controlled 00 R~elease, supra, vol. 2, pp. 115-138 (1984). Preferably, a controlled release device is Ct latroduced into 4 subject in proximity to the site of inappropriate, immune activation or a Dimor. Other controlled release systemns are discussed in the review by lAuiger (Science 249:1527-1533 (1990).

INDThe pharmaceutical preparation can comprise the selective androgen receptor (Ni modulator alone, or can further include a pharmiaceutically acceptable career, an4 can be in solid or liquid form such as tablets,* powders, capsules, pellets, solutions, suspensions, 00elixirs, emulsions, gels, creams, or suppositories, including rectal and urethral suppositories. Pharnaceutically acceptable carriors include gazzts, staches, sugars, cellulosic materials, and mixtures thereof. The pharmaceutical1 preparation containing the selective androgen receptor modulator can be administered to a subject by, for example, subcutarivous implantation of a pellet; in a further embodinrt the Pellet provides for controlled release of selective androgen receptor modulator over a period of Is time. The preparation can also be administered, by intravenous, intraarterial, or intramuscular injection of a liquid preparation, oral administration of a liquid or solid preparation, or by topical application. Administration can also be acomplizhe4 by use of a rectal suppository or a urethral suppository.

The phatmaceutical preparations of the invention can be prepared by known dissolving, mixing, granulating, or tablet-forzring processes. for oral administration, the selective androgen receptor modulators or their physiologically tolerated derivatives such as salts, esters, N-oxides, and the like are mixed with additives customary for this purpose, such 49 vehicles, stabilizers, or Inert diluents, and converted by customary methods into a suitable form for administratior4 such as tableis, oated tablets, hard or sot gelatin capsules, aqueous, alcoholic or oily solutions. Examples of suitable Inert vehicles are conventional tablet bases such as lactose, sucrose or cornstarch in combination with binders like acacia, cornstarch, gelatin, or with disintegratig agents such as cornstarch, potato starch, alginic acid, or with a lubricant like Btearto acid or magnesium stearste.

Examples of suitable oily vehicles or solvents are vegetable or animal oils 00 such as sunflower oil or fish-liver W. Preparations can be effeoted both as dry and as wet grariules. For parcnteral administratiou (suboutananus, intravenous, intraarterial, or intramuscular injection), the SARM agents or the non-steroidal agonist agents or their physiologically tolerated derivatives such as salts, oatens, N-oxides, and the like are S converted into a solution, suspension, or emulsion, if desired with the substances IND customary sAn suitable for this purpose, for example, solubilizers or other auxiliaries.

Examples are: sterile liquids such as water and oils. with or without the addition of 4 suirfactant and other pharmnaceutically acceptable 4duvants. Illustrative oils are those of 00 petroleum animal, vegetable, or synthetic origin, for example, peanut oil, soyban oil, or mineral oil. In general, water, saline, aeous dextose and related sugar solutions, and glycols such as propylene glycols or polyethylene glycol ane preferred liquid carriers, particularly for injectable solutions.

The preparation of phannacentical compositions which contain an activo component is well understood in theaaL Typically, such compositions arm prepared as aerosols of the polypeptide delivered to the nasopharynx or as injectables. either as liquid solutions or suspensions, however, solid forms suitable for solution in, or suspension in. liquid prior to injection can also be prepared. The preparation can also be ernulsifiec. The active therapeutic ingredient is often mixed with oxcipients that ane pharmaceutically acceptable and compatible with the active ingtedient. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, or the like and combinations thereof.

In addition, if desired, the composition can contain milnor amounts of auxiliary 2z substances such as wetting or emulsifying agents, pH buffering agents, whirch enhance the effectveness of'the active ingredient An active component cm be fominulated into the composition as neutrali2d pharmaceutioally acceptable salt forms. Pbarmaceauticafty acceptable salts include the acid addition salts (famned with the free amino groups of the polypeptide or antibody molecule), which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids 4s aqetic, oxaio, tartarlo, mandelic, and4 the like.

00 Salts formed from the free carboxyl groups can also be derived from inorganic bases such as, for cxample, sodium, potassium, amnmoniumn, calcium, or ferric hydroxides, and such organic bases as isopropylarnine, trirnethylarnine. 2-ethylarnino ethaol, histidine, procaine, and the like.

IND For topical administration to body surfame using, for example, creamns, gels.

drops. and the like, He SARM agents or the non-agonis steroidal compounds or their physiologically tolerated derivatives such as salts, esters. N-oxides, and the like are 00prprdadapidasouinssesosoreusosiaphsooial 0rprdadapida ouinssesos reusosi h~ooial 0 10 acceptable dilueut with or without it pharmacoutical carrier.

In another embodiment. the active compound can be delivered in a vesicle, in particular a liposome (see Langer, Science 249:1527-1533 (1990). Treat at al., in Liposornes in the Therapy of nfetious Disase and Cancer, Lopez- Berestain. and Fidler is Liss, New York, pp. 353-365 (1989); tLopez-Beresteiri, ibid., pp. 317-327; see generally ibid).

For use in medicine, the salts of t SARM or the non-steroidal agonist compounds will be pharniaceutioally acceptable salts. Other salts may, however, be usefurl in the preparation of the compounds according to the invention or of their pharmaceutically acoeptable salts. Spitable pharmaceutically accptable salts of the compounds of this invention iuclude acid addition salts, which may, for example, be formed by mixing 4 solution of the compound according to the invention with a solution of a pharmuccuticalUy acceptable acid such as hydrochloric acid, sulphuric acid, 2S methanesulphonic acid, fmwrio acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic aoid, citric acid, taraic acid, carb~onic acid or phosphoric acid.

The present invention furher relates to a method of determining the presence of a selective anidrogen mnodutor compound and/or at non-steroids I agonist comnpond of the preent invention in a sample. The method comprises the steps of obtaining the sample, and detecting the compound In the sample. thereby determining the presence of the compound in the sample.

00 In one embodiment, the sample is 4 blood serumn sample. in another embodiment, the sample is a plasma samnple. In another embodiment, the Sample is 4 W)ine sample. In another embodiment, the sample is a safiva sample. In another S embodiment, the sample is any other tisue sample.

In one embodiment, the detection step comprises measuring the absorbance of the compound at a prodetenmined wavelength. For example. the compounds of the 00 present invention absorb in the ultraviolet region of the spectrum, with an absorbency to peak at 270 nin. Thus, ina one embodiment of the present Invention, the compound is detected by monitoring the UV absorbance of the sarnple at 270 imn. It should be noted that the present invention is not limited to UV absorption, and that any other spectrometric methods of identification are applicable. F~or example, compounds can be detected by measuring their idfr-red or visible absorbanoa.

In another embodiment, the present invention ftither provides a method of doterxninizng the concentration of a selective androgen receptor modulator comipound 4ud/or a non-steroidal agonist compound of te present invention in a sample. The method comprises the step of obtaiuing a sample: determfinin the level of the compound in the sample, and calculating the concentration, of the compound in the sample by comparing the level with a standard sample containing a known concentration of the compound. Caliration curves of known concentrations of the compound in the sample, can be obtained, and the concentration of the compound in the tast sample is Calculated therfrom. By 'level" It is meant the absorption level of the compound at the MeaUred Wavelength.

Ina another embodiment the compound is detected in the sample by contaeting the sample with a binding protein which qpeifically binds to the compound, and determining the amnount of binding protein bound to the compound. The concentration of the compound can be determined by measuring the amount of binding protein bound to the compound, and comparing that amount to a standard sample containing a knows concentration of the compound binding protein omiplex.

00 Protein levels can be determined. according to standard techniques, as described in Sambrook at al. Briefly, 4 sample obtained from it subject is contacted with CI a binding protein which specitlcally binds to a specific compound of the present S inventi, and the amount of complex formed between the binding protein and the IND compound is determined. In one embodiment, the binding protein Is an antibody which specifically binds to one or more compounds of the present inven~tion- In another embodiment the binding protein has a detectable label bound thereto, and the complex 00 between the binding protoin-label compund is determined by visualizing the complex.

As defined hmrin, "contacting" momns that the binding protein is intlroduced into the sample in a test tube, flask, tissue culture, chip, array, plate, microplate, capillary, or the like, and incubated at a tempertur and time sufficient to permit the binding componeot to bind to a cell or a fraction thereof or plasra/serum or a fraction is thereof containing the target Methods for contacting the samples with the binding proteins, or other specific binding components are kntown to those skilled in the ant and may be selected depending on the typo of assay protocol to be run. Incubation methods are also standard and are known to those skilled in the art.

"Viva~izing" the complex my be carried out by any means known in the art including, but not limited to, ELISA, radioimnmunoassay, flow oytometry, dot blots, western imimuinoblotting combined with gol electrophoresis, jmmunohistochernistry at light and electron pe levels, HPWC and mass spectrometry.

B3ither monoclonal or polyclonal antibodies (as well as any recombinanit antibodies) specific for the selootive androgen modulator compndOiCs or the nonsteroidal agonist compounds of the present invention can be used in the various imunoassays. The antibodies may be deteotably labeled, utilizing conventional labeling techniques well-known to the art~ As used herein, the terma "label" refers to a -molecule, which may be conjugated or otherwise attached covalently or noncovalently) to at binding protein as defined herein. Labels are known to those skilled In the at. Thus, the antltdies may be labeled with radioactive isotopes, non-radioactive 00 isotopic labels. fluorescent labelS, enzyme labels, cherniluminescent labels, biohmainesceftt labels, free radical labels, or bacteriophage labels, using techniques known in the amt EXaMtPleS Of radioisotopic labels are .sup.3 .sup. 125 1, -sup. 131 1, S, .Sup. 14 C. ae. Examples of non-radioactive isotopic labels are .sup.55 Mn.

IN 5 sup.56 Fe, etc. Examples of fluorescence labels are fluorescent labels which are directly labeled with the preferred fluorescence label, or fluorescent labels which are CI indirectly labeled with the preferred fluorescence label. In the la&t case. the preferred fluorescence label Is conjugated to a secondary antibody, which is directed against the 00 first antibody, such as an anti species Ig antibody. Typical fluorescent labels; include, but are not limited to a fluorescein label, an isothiocyanate label, a rhodamine label, 4 phycoerythrjn label, etc., for example fluorescein isothiocyanate (PITC, International Biological Supplies, Melbourne, FL). rhodamine, phycoeryhin Coulter Corp., Hialeah, phycocyanjn, alophycocyanin, phycoeryalin-cyanin dye 5 Coulter), label, a phycocyanin label, an allophycocyanin label, an 0-plithaldehyde label, Is a fluorescarnine and Texas R.ed.

Examples of enzyme labels include alkaline phosphatase, beta-galactosidase, glucose-6-phosphate dehydrogenase, rnaleate dehydrogenase, and peroxidase. Two principal types of enzyme immunoassay are the enzyme-linked immunosorbn assay (ELISA), and the homogeneous enzyme inmmunoassay, also known as enzyme-multiplied immunoassay (EMIT, Syva Corporation, Palo Alto, CA).

In the ELISA system, separation may be achieved, for example, by the use of antibodies coupled to a solid phase. The EMIT system depends on deactivation of the enzyme in the tracer-antibody complex; the activity can thus be measured without the need for a separation step.

Particularly sujitable labels include those, which permit analysis by flow cytometry. fluoroolwomes. Other suitable detectable labels include those Useful In colorimetric enzyme systems, e. horseradish peroxidase (1WU) and alkaline phosphatase Other proximal enzyme systems are known to those of skill in the aWt including hexolcinase in conjunction with glucose-6-phosphate dehydrogenase.

00 Additionally, chemninescent comnpounds may 130 used as 1abe16.

jhaiumineccit labels. mnob as green fthomecct proteins. blue fluoreseent preteins.

=4d vuriants theof ame known. Also biouincacence or ohatailuadftescence oa= be deteted using, rspeciively, NAt) oxidoroductase with luciferue and substrates NAX)1 and WN or peroxidase'with luminol and substrate peroxide. Typical ohOMihrUueWoet compounds include knuteol, isoluminol, aromnatic aoridinium caters, irnidazoles.

acridinum solb, and oxalate esters. Similarly, bioluminescent compounds MaY be uitlized for labelling, the blohaxninascent compounds Including luciferin, lUCifrase, and 00 a~equorin. Owce labeled the antibody may be employed to identify and qunif immunologic counterpamt (antibody or mntigenic polypeptide) utilizing techniques CI well-knowu to the art The following examples are presented in order to more fally ilustrate the prefarred embodiments of the invention. They should in no way be construed, however, as limting the broad scope of the invention, EXMEIMENTAU DITAILS SEMTON Nonsteroldal LUganda with Androgenic and Anabolic Activity The SARM compounds provided haremn wee designed, syatheized anid evaluated for in-vitro Anid in-vivo pharnacologic activity. The in-vitro androgen receptor binding affiity anid ability to maintain andirogeni dependent tisage growth in castrated animals was studied. Androgenic actiVity was Wonitored as the ability of the SARM compounds to mintain and/or stimulate the growth of the prostate and semiinal vesicles. as measurod by weight Anabolic: activity was monitored as te ability of the SARM compounds to maintain and/or stimue tba growth of the lavotor ani muscle. as measured by weight- 00 Synthetic ftocdnu g~f Compound$ IVIZ!: (2R)-1MafcloylpyrotIiO-2-rbOXqli Acdd (R-129). D-?roline (R- 128, 14.93 g, 0. 13 mol) was dissolved in 71 niL of 2 N NaOH and cooled in an ice bath; the resulting alkaline solution was diluted with acetone (71 wL). Am acetone solution (71 INDmL.) of metacryloly chloride 127 (13.56 g, 0.13 mot) and 2.N NaOH solution (71 rnL) Wore simultencously added over 40 min to the aqueous solution of D-proline in fin ice batiL The pH of the mixtre was kept at 10-1 IOC during the addition of the motacrytoly chloride. After stining (3 h, room temperature), the mixture was evaporated in vacua at 0010 a temperature at 35-45 C to remove acetone. The resulting solution was washed with ethyl other and was acidified to pH 2 wih concntate HC. 7Ue acidic mixtur was saturated with NACI and was extracted with EtOAc (100 niL x The combined Wdacts were dried over NA2S0 4 JUrerd through Celito, and ovaporated in vacua to give the crude product as a colorless oil. 19.eryta~lization of the oil from ethyl other and homares afforded 16.2 of the desired compound as colorless crystals: mp 102-103 *C (lit [2141 nip 102.5-103.5 0 the NMR spectrum of this compound dernonstrated the existence of two rotmers of the title compound. 'H NMR (300 MHz. DUSO-4l) 855.28 4n4 5:15 for the first rotantor, 5.15 and 5.03 for the second rotamer (totally 2H for both rotamers, vinyl CR 2 4.48-4.44 for the first rotsxuir, 4.24-4.20 (in) for the secod rotainer (totally lH fbr both roturners, Cti at the chkial cantor), 3.57-3-38 (mn 2H, CH3), 2.27-2.12 (1K-i CM, 1.97-1.72 (mn, 6H1, CU 2 ,'CR Me); 'C NM (75 bMz, l)MSO-4) 8 for major rotamner 173.3. 169.1, 140.9, 116.4. 58.3., 48.7, 28.9, 24.7, 19.5: forrminor rotftmor 174.0, 170.0. 141.6, 115.2, 60.3.4S.9, 31.0.22.3, 19.7; MII (KBr) 3437 1737 1647 (CO, COOR), 1584, 1508, 1459, 1369, 1348, 1178 orn'l (416o 26 +8Q.8* 1, MeOR); Anal. Calod. for CgU))NOS: C 59.00, 11 7.15S. N 7.65.

Found: C 59.13, H 7.19, N 7.61.

(3,a)3Hooebk-eblUrhdoproo21 cIII,4Joxazka.-1,4-dlone (RI, R-130). A solution of NBS (23.Sg, 0.132 mol) in 100 cit of DMF was added dropwise to a stirred solution of compound R-229 (16.1g, 88 mmcol) in 70 mL of V)MF under argon at room temporatdro, and the resulting mbxtur 4as stirred 3 days. The solvent was removed i vacua, and a yellow solid was precipitated.

00 The solid was uuspea4ed in water, stirred overnight at room temperature, Atered. and died to give 18.6 (smaller weight when dried 34%A) of the title compound as a yellow soU- mp I1S2-154 OC (lit (2141 ap 107-109 QC for the S-isomer); IH NM~R (300 MHz. DMSO-d 6 8 4.69 (44, J 9.6 Hz, J 6.7 Wz I H, CIt t the chiral center). 4.02 S(d. J 11.4 Hz, I H, CM~, 3.86 J~ -1I.4AHz, I H, CTHb), 3.53-3.24 (m4H, CH 2 2.30-2.20 (in, I H. CIU) 2.04-1.72 (mn 3H, CH2 andl 1.56 2K, Me); NIR hz14 DM80.4) 8 167.3, 163.1, 83.9. S7.2, 454, 37.8. 29.0, 22.9. 21-6; ZR 3474, 1745 1687 1448, 1377, 1360. 3308, 1227. 1159. l062cmh-': fabD'+124.5 00 (c chloroform); Anal. Calcd. for CgT, 2 BrN03.: C 41.24, H 4.61. N 5.34. F~ound: C 41.46, H4.64, N5.32.

(2R)-3-Braimo2bydroxy-2-DthyIprOPeflo1c Acid (Rt-13 A mixture of bromolactoue R-130 (I18.5g. 71 romol) in 300 roL of 2.4% EH~rwas heated at retu'i for I h. The restulting solution was dilutedl with brine (200 mL, and was extracted with ethyl acatate (100 mb x The combined extracts were Washed With saturted NSH*C03 (100 mL. x The aqueous solution was acidified with concentrated lId to PH which, in tWin, was extacted with ethyl acetate (100 tub x The combined organic solution was dried over Na2SO4, filtered through Celite, and evaporated in vaouo to dryneas.

Rerytal~ization, fron toluene afforded 10.2 g of the desired comipound as colorless mytls: Top 107-109 9C (Ot. (214] mp 109-113 OC for the S-isomer); 'H NMR (300 Miz, DMSO-4) 6 3.63 J 10.1 Hz, lH, CHH.), 3.52 J 10.1 Hz, 13*, CWQb, 1.35 3H. Me); MR (K.Br) 3434 (0OH), 3300-2500 (C00OK) 1730 1449, 1421, 1380. 1292, 1193, 1085 6+ 10.50 (c 2.6, MeOli); Anal. Calod. for C.11 7 B3: C 26.25.,1*3.86. Fonid: C 26.28, H 3.75.

N-4Nto4rfurmtyptyl(R--rm4hdoy2 tMtbYlproIanavulds (R-132). Thionyl chloride (8.6 g, 72 mrool) was added dropwiso under argon to a solution Of brOmoatid R-131 (11.0 g, 60 MumoQ in 70 niL of D)MA at to -10 C. 71e resulting mixtur was stined for 2 hi under the same conditions. A solution of 4-uftro-3-trjfluoromathyl-anllke (124 g& 60 minol) in 80 znL of DMA was added dropwise to the above solution, and the resultS mixture was strd overnight at roI= topOrAtliro. The solvet was removed On RotaVapOr Using high vRCUUM Oil pumIp; 00 the residue was diluted with Bauxrate4 NaHCO, solutioni, end extracted with ethyl other (109 niL x Combined mitracts were dried over mnhydrous NkSO 4 fitered throu~gh Cellie, and pinified by flash cbremtograpy on silica get, using mnethylene chloride as c-Ichantf to afford 18.0 g (110%) of the desired compoud:- mp 98- 100 *C (Rr 0.2 silica gel, CH2CI2); 'H NMR (300 1INMz DMSO-4) 8 10.54 I H, NHl), 8.54 (4l, J -2.1 Hz, 1KU! ArH), 8.34 (4d, J 9.0 11z, J -2.1 Hz, 1K, ArH), 8.18 (Cl, J3 9.0 Hz, IH. ArH).

C1 6-37 1K, OH), 3.82 (di, J 10.4 H2, 111, CWH.), 3.58 J -10.4 H7, lH, CHHW,.

1.48 3K, Me); "C NqMR (75 MK2, DMSO-Q. 173.6 143.0, 127.2, 123.2, 00 122.6 33.0 Hz), 122.0 (cj, J3 27 1.5 Hz), 118.3 J 6.0 74.4, 41.4, 24.9; IP.

(KBr) 3344 1680 1599, 1548 (Cu-C. Ar), 1427. 1363, 1161 MS (ESi): ni/z 370.8 Anal. Culod. for CIIHIBrN 2

O

4 C 35.60, H 2.72, N 7.55. Found: C 35.68, H5 2.72, N 7.49, N-4nto3tiloaehohull($-- aeyan~hnxl2 bYdroxy-2-incthyipropsaaide (S-147). The title compound was prepared. fr compound R-132 (0.37 g, 1.0 nunol), 4-acetazidaphanol (0.23 1.5 mmrol) X2CO3 (0-28 s. 2.0 minol), and 10% of beuzyltributylauamoniuni chloride as a phase tmanafr catlyst in 20 reL of methyl ethyl kotone was heated at reflux overnight Under argon.

The reaction was followed by TLC, the resulting mixture was filtered through Celite, anld concentrated in vaua to dryness. P~urification by fah column chrmatogmnpy on Silica gel (bexanes-ethyl acette, 3: 1) yWeldd 0.38 g (Rr =0.18S hoanes-ethyl acetate, 3:1) of the 4esix-ed compound as a light yellow powder. np 7"-4 9C; The solid can be recrystaized from ethyl acetat* and hexane); 1H NMP. (300 M-z, DM50-dd 8 10.62 1H, NM, 9.7S IN, 8.56 1.9 Hz, 1k!, Ark!), 8.36 (cid, J3 9.1 Htz, J3 1.9 Hz, 1K. ArlI), 8. 18 J 9.4 Hz, IH, ArK!) 7.4S-7.42 (in, 2H!, Ark!, 6.85-6.82 (v4, 2H1, Arft) 6.25 111, OR), 4.17 J 9.5 Hz, I H, CMI!., 3.94 J3 9.5 Hz, IN, CUR,. 1.98 3H!, Me), 1.43 31j, Me); '1C NMR (75 MHz, DMS0-dO 8 174.6 167.7, 154.2, 143.3, 141.6, 132.8. L27.4, 123.0. 122.7 J 33.0 Hz), 122.1 (q, J -271.5 Hz), 120.1, 118.3 I 6.0 Hz), 114.6, 74.9. '73.9, 23.8. 23.0; IR(00r) 3364 1668 1599. 1512 Ar), 1457, 1415, 1351. 1323, 12.39, 1150 1046 cal t MS CWS!: rn/a 464.1 t Anal. Calcd. for CigHAPeiW: C 5 1.7 1, H 4.11, N 9.52. lPotmn4 C 52.33, H 4.40, N 9.01.

00 The in-vitro activity of the SARM compounds speCifically compound VII, 0010 demonstrated high androgen receptor binding afflunity (KI 7.5 nM). Animal studies with the SARM compounds, speoifically compound V. demonstrated thai it is a potent androgenic and anabolic nonsteroidal agent Four groups of rAts were used for these studies: intact controls, castrated controls, catrated animals treated with testosterone propiouat (100 pg/day), and castrated animals treaed with compound is V (1000 14g'day). Testosterane and compound VII were delivered at a constant rate for 14 days via subcutmaneus osmotic pumps.

The results of these studies are shown in Figure 1. Castration significantly reduced the weight of androgenic prostate and seminal Vesicles) =an raolic levator Wm muscle) tissues, Win had little &efct on animal body weight Treatmt.

of castrated 4uWrpas with testostarowe propionate or compound VUI maintained the weight of androgenic tissues to the samie degree. Compound VII Wa similar androgenic activity as testosmuone propionate the prostate and semina vesicle weights ware the aanMO), but much greater eocazy 4s an anabolic agent. Compound VII showed greater 40abolic activity thm testosterone proponat 4t the doses tested the levator ani muflscle maintained the weight 4s iptact control animals and was greater than that observed for testostere). The experiment presented herein ame the fist ina-vivo results which 4emonsu-Mt tissue-aelactive axxtrwogenlc and anabolic activity differing androgenio and anabolic potancy) of a nonawreidai Uigand for the andr-ogen receptor, 00 Nonsteroidal Ligands with Alsdrogenic and Anabolic Activity te in-vivo efficacy ad acuflQ toxicity of four novel uinmmoidal androgens (compounds IV, V, V1 and VUI) in rats was exanined. In-vitro assays establihed that these compounds bind the adrome receptor with very high affinity. The structume mzd 'Ramos of the four compounds are presented below: Crrx-014 (M~-015 GTx-016 GTX-017 R=COCH3

R-COC

2

HS

R-NHCOCIHa EXPEIUNTAL METHODS Materials. noe S-isomers of compounds arx-014 (compound GTX-015 (compound GTX-016 (compound VI) and eU007 (Compo~and VUI WherCIP R is NHCOCHi3) and the R-isomar of GTx-014 were synthesized in accordance with the scheme as set fw*t in Figure 9. Teestorone pioate Mr), polyethylene glyr-ol 300 (PRO300, reagent Srade) and neutral buffared torrualin (101/ wlv) were pwurasd from 00 Sigm~a Chemical Company (St Louis, MO). Aizet osmotic pumps (model 2002) were purchased homz Alma Corp. (Palo Alto, CA).

Animals. Immature male Spregue-Dawley rats, weighing 90 to 100g, Were Spurchased from Harlan Bioscjeuce (ludianapolis, IN). The animals were maintained on a 12-hour light-dark cycle with food and water available ad libim.. The Wmflal Protocol was reviewed and approved by the Iitutional Laboratory Animal Care and Use Committee.

00 1 Stu~dy Des'gL Rats were randomlly distributed into twelty- ie (29) groups, with animals per group. Treatmeont groups are described in Table I. One day prior to the start of drug treatment, animals in groups 2 through 29 were indvidually removed from V4e Cage, weighed andi anesthetized with an intrearitoneal dose of ketemin& yla ine (87113 rag/kg; approximately I mL pe kg). When appropriately anesthetized no repose to tot pinch), the animals' ams were marked for identification purposes.

Animals were then Placed on a sterile pad and their abdomen and scroturn washed with betadine and 70% alcohol. The testes were removed via a midline smrotal incision. with sterile, auture being used to ligate supre-testicular tissue prior to surgical removal of each tests. The surgical wound site was closed with sterile stainless steel wound cliPs. and the site cleaned with betailine. The animals were allowed to recover on a sterile pad (unil! able to, stand) and then returned to theair cage.

Twenty-four hors late, animals in groups 2 through 29 were re-anesthetized with ketumine/xylazine, ad an Alzet osmotic pump(s) (model 2002) was placed suboutaneouly In the scapular region. In this instance, the scapular region was shaved and cleaned (betadine and alcohol) and a small incision (I cm) made using at stexile scalpel. The osmnotic pqmp was inserted and the woxnd closed, with a sterile stainlss steel wound clip. Animals wern allwed to recover and were returned to their cage.

Osmotic pumps omtaind the appropriAte teet (designated in Table 1) dissolved in polyethylene glycol 300 (PB0O300). Osmotic pumps were Riled with the appropriAte solu~tionI one day prior to implantasion. Animals were monitored daily for signs of acute toxicity to drug treatment lethargy, o coat).

00 After 14 days of drug trearrnent, rats wene anesthetized with ct ketamnchyla no. Animals were then saerilced by exsanguinations under anesthesia.

A blood sample was collected by venipunoture of the abdominal aorta, and submitted for N acmplet blood cell analysis. A portion of the blood. wats placed in a separate tube, centrifuged at M~OOOg for I inntc, ad the plasma layer removed ad fOzen at -20 0

C.

The vental prostates, semin~l vesicles, levutor ani muscle, liver, kIdneys, &pleon. Lungs N and heaut wowe removed, cleared of extraneous tisane, weighed, and placed in vials containing 10% neutral buffered forroalin. Preserved tissues were seat to OTX, Inc. for 00 bistopathologiCal 4nalysis.

For data analysis, the weights of all organs were normalized to body weight, and analyed for any statistical significant differnce by single-factor ANOVA. The weights of prostate and seminal vesicle were used 49 indexes for evalation of androgenic activity, and the levator ani muscle weight was used to evaluate the anabolic, activity.

REULTS

The androgenic and anabolic activities the S isomers of compounds CITX-014, OT)x-015. GTX-Ol6 and GQx-007, and the R isomer of GTx-O14 were examined ip a castmatd rat model after 14 days of administration. Testosterone propionate, at inceasg doses, was used as the positive control of anabolic and androgenic effects.

As shown in Figures 2 and 3. the weights of prostate, seminal vesile, amd levator ani muscle in castrated. vehicle-treated rats decread significantly, due to the ablation of endogenous endrogen production. Exoginous administaton of testostmee propionate. in androgenio and anabolic steroid, increased the weights of prostate, seminal vesicle. and levator nii muscle in castrated rats in A dose-depandeot manner.

7Ue R-isomer of OTx-O14. and S-isomer of OTz-0S Sand GT%-016 showed no effect on the weights of prostate. seminal vesicle, and levator ani muscle inl castrated animals' (data not shown). The S-isomers of GTx-007 (Figure 2: S-GTx-O0') and GT%-014 (Figure 3: S-GTx-0) 4) resulted in dose-dependent increases in prostate seminal vesicle and levator miii muscle weights. Compared with testosterone. propioft, S-C1TX-07 00 C~1 showed lower potency and iatiinsic activity in Increasing the weights of prostate and Ct seminal vesicle, but a greater potency and intrinsic activity in incresing8 the Weight Of levator ani nwasle. Patioulszrly, S-GTx-007, at a close as low as 0.3 rag/day, was able to maintain the levsar ani muscle weight of castrated animals in the sanme level As that Of intact animals. Thus. S-QTx-007 is a potent nonsteroldal anabolic agent with less IND androgenic activity but morm anabolic activity than testosterne propionate. This is a signlftcapt imnprY=uet over previous claims, in that this compound selectively c"1 stimulates muscle growth and, other anabolic effect while baving less effect on the c-i prostate and seminal vesicles- This may be particulaly relevant in aging men with 00J 0 conlcerns related to the development or progression of prostate cancer.

QTX-014 was lIss potent than GOx-007, but showed greater tissue selectivity (compare effects on t prostate and seminal vesioles in Figures 2 and GQx-014 significantly increased levator aW mauscle weights, but showed little to no ability to 1S stizniate prostate and seminal vesicle growth the prostate and sezninal vesicle weights were less tha= 20% of that observed in intact animals or in animals treated with testosterone Propionate).

Itesults showed that none of the eammined compounds produced significant effect on body weight or the weights of other organs liver, idneys, spleen, lungs and heart). Nor did say compound produce any signs of acute toxicity, as gauged by diagnostic hemAtology tests and visual examinaton of animals receliin treatmenfts.

Importantly, GTx-007 did not suppress the production of luteiniing hormone (LH) or follicle stimulating hormone (FSH) at a dose of 0.3 mg/day a dose that exhibited maximl anaboic effects).

In summnary, S-GTx-007 exiulsitad exceptional anabolic activity in animals by Maintaining the weight of levator ani wascle after removal of enogenou~s androgen.

This discovery represens major progress towards the development of therapeuatically useful nonsaroidai mndrogens, and a major improvement (Lae., tissue selectivity and potency) over previou~s drugs in this class. S-GTx-014 and S-GTx-007 showed selective anabolic acivity in comparison with tetosterone prOpionste, an anIdrogenic and anablic 00 steroid. noe tsuo-electivo activity is actually one of the advantages of nonateroidal Ct androgens in terms of anabolic-rolatod applications.

*Despte simlarities in stcture and in-vitro Awntional activit:y, the S-isomers of compounds QOTh-014, GIN-015, 07'x-016, and OT%-007 exhibited prafound IDdifferences in tenn of their in-vivo activitr. M~-007 the miost cftlicacious androgenic and anabolic activity in animals. with the anaboflc activity greater than that of testosterone propionate. GNx-014 showed a smll degree of androgenic activity. but an 00 anabolic activity comparable to testosterone propionate. in contrast, GTx-015 and GTX- 016 failed to produce any 4ndrogeaic or anabolic activity in-vivo.

Tese studies show the discovery of two members (GQx-014 and OUx-007, compounds, compounds Rl and V respectively) of a new class of selective -androgen receptar modulators (SAIRMS) that demonstrate potent anabolic; effects muscle growth) with less androgenic activity prostatic growth). This new class of drugs has several advantages over nou-selective androgens, including potential therapeutic applications in males and females for modulation of fertility, erythropoiesis, osteoporcs, sexu4I libido and in men with or at high risk for prostate cancer.

Further, Figures 7 and 8 denzonstrate the effects of GOx-014 and GTx-007 on LH and FSH levels in rats. Thoe res-alls fuarther demonstrate the novelty of these SARM9, due to their differential effects on these reproductive hormones, thus dernonstrating the tissue-specific pharmacologic; activity. In Figure 7, LH levels in castrated animals treated with TP and OTX-014 wer significantly lower than those of untreated anmals castrated controls) at doses greater than or equal to 0.3 mg/day.

However, higher doses 0.5 mg/4ay or higher) of GTx-007 were required before significant decreases in LH levels were observed. T7hus, O N-007 does not suppres LH levels at doses that arm capable of elicitin maximal stimulation of levtor ani muscle growth, In Figure 8, PSE levels in castrated animals treated with GTx-O 14 were significantly lower than those of untreated aniuMs (iLe,, castrated controls) at doses of mg/day or higher. Similarly, lower FSH levels were observed in anirnals treated with TP. H{owever, only this diffamoue was ouly significat at a dose of 0.75 mng/day. PSH 00 00 levels in saikas treated with GUi-007 w=r not zM~c~iiy differet fom those of uated anima s at any dose level tstd. Thus, MY-007 does not suppress FSH levois at doses that are caspable of elioit maximal stinulrlcnof lovator ani muscle gm~wth.

Table 1. Animals Groups nd Experimental Dosig Group casftritaed? m -Dose of sonals I wo None None T 2 Yes -Nn Vehicle only 3 Yes esti ___ay Yes Testosterone 03m/a 6 -ye Testostemoe 0.7 i ay 00 INOEXM PLE 3 ci Pburmsaonetics of GTx-007 In Dogs 00 10 The paackntcs of S-GTx-(Y7, a novel selective androgcr receptor modulator (SARK, ware characterized in' beagle dogs. A folur-treatmcnt, four-period crossover design was utilized in the axidy, which involved a total of six beagle dogs, three of each, goder. Bach animal received a 3 raS~g) IV dose, a 10mrg/kg TV dose, a mng/k P0 dose in solution, ud at 10 mg/ks PO dose in capuie, ini a randomly ussigp.4 order. Then was an oine-week washout period between treatmients. Plasma samples Were colemted for up to 72 hr afte drug aftisbtrioD. Plasma S43-00-O7 coflantlattons were analyzed by 4 validated EPLC methiod. The clearanice volume of 4istribution (Yas), half-life (Tita, and other~ pbarmacokinctic puamotn wae deteniaiuod by nocozmparbneatal moathods. Results showed that S-Gnx-007 was cleared from dog plasm wit a teruimW Top of about 4 hr and a CL of 4.4 mL~rzniafg affter IV adinisuton. Figures~ 4, 5, and 6 show V1ie plasma conoratio-time profiles of S- G7 x-007 after admin~istraution of ma intlravenou3s solutIoA, oral solution, and oral capsule, respectively. The pharmacoldnatics we=e dose- and Sendor-indepndant. The oral bioavailability of S-O~x-007 varied wit the dosage formi, anid averaged 3 9% and I -W for solution and capsule, respectively. Thus, S-CSTx-007 demonstrided mioderate halflife, slow clepwce and modwate biw~ailability in~ beagle dogs, Identifyinig it as the first of 4 new clss of aniiy bio~v4Uble tissue.selactive androgen receptor modulators.

EXAAVLR 4 g~X-OO Auulysta bY HFLC A reversed phase bigh preuum liquid chjromuuagraph (HPLC) assay wa 00 dervelopedl to quanttata GTx-007 concentrations In dog plasma. Dog blood samples wr obtained by veaipnnctne and centrifaged tit I OOOg for IS minutes. Samnples were stored fioze at -20*C =61i analysis. Individual samples wert rapidly thawed and an aliquot ml) was spaked with internal standard (20 ±Ll of a 200 pag/ml aqueous solution of 5CM-1U-87). An aliquot of 1 ml of acotonitrile was added to the samples to precipitate plasmA Proteins. The samples were vortaxeci and then centrifuged at 10OOg for minutes. The supernatait was deated! into glass extraction tubes and 7.5 ml of ethyl acetate was added. The extraction mixture was left at room tempealre for 2.0 minutes, 00 and Vortexed several times during this interval. The samples were them centrifuged at 1000g for 10 minutes, and the organic phase was removed and placed in conical- C~1 bottomned glass tubes. The organic phase was evaporated uder nitrogen. The samPles Were reconstituted in 200 p1 of mobile phase (35-65 ace rilev~wac) and transferred to an antosapler vial for HPLC injection (Waters 717 plus autosampler, Waters Corp., Milford. MA). The isocratic mobile phase of 35% acetenitie, in water was pumped at 4 flow rate Of 1 mI/min (Model 5 10. Waters Corp.). The statonary phase was a CIS reversed phase column (Novapak C 19, 3.9 x 150 mm). Analytes were monitored with UV detction at 270 rim (Model 486 absorbance detector, Waters Corp.). Retention times for GTx-007 and CM-IT-97 were 11.)1 and 16.9 rairmls, respectively.

Chromatography data was collected and analyzed using Millennium software. Plasma COnceftatioD of CITX-007 in each sample were dottiin by comparison to calbration curIves. Calibrtin curves were constmuoed by oddig kaown amounts of GTx-007 to dog plasma. Final GTx-007 concentrations in dog plasma samples Maed in the ca][ibration cines w=r 0.08, 0.2, 0.4, 2. 4, 10, and 20 ig/1 Calibration curves Were liner over this concentration range and eychibited corretatlon Coeffioients (r2) of 0.9935 or greater. Intra- and ipte-day coeffcients of vuriton for the standards ranged fromn 6.4% for 0.08 pig/rl to 7.9% for 20 pglml.

Melting points were determined on a Thomas-Hoover capillary melting point apartu and ane uncorrectd. Infrred spectra were recorded on a Perkin Elmer System 2000 Pr-IR. Optical rotations wene determined on an Autopolo MI Automatic Polarimaer (Rudolph Research Model M-589-10. F~airfield. Now JerseY). PrOton and carbon- 13 wagnetic resonance spectra were obtaiited on a lnzke AX 300 spectninietor 00 (300 and 75 Mhiz for 1H{ and 3 C, respectively). Chemical shift values were reported as Ct parts per raillion relative to tatrmevtbylsilaxe CTMS). Spectria data were consistent with aBsigDed structures. Mvass specr were determined on 4 Bmkocr-1HP Esquire LC Systom. Elemental analyses were performed by Atlantic Microlab Inc. (Norcross. GA), s and fousnd values were within 0.4 of the tearetical values. Routine thin-layer chroresiogray (TLC) was performed on silica gel on aluminume plates (silica gel 60 F 254, 20 x 20 cm, Aldrich Chemical Company Inc., Milwwuke. WI). Flash chromatography was performed on siica gel (Merclk, grade 60, 230-00 mesh, 00 Tetrahydkofirran (TfW) was dried by distillation over sodium metal. Acatonituile (MoCN) and mnethylene chloride (CU 2

CI

2 were dried by distillation from P 2 0 5 In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word "comprise" or variations such as "comprises" or "ccomprising" is used in an inclusive sense, i.e. to specify the presence of the 1 5 stated features but not to preclude the presence or addition of further features in various embodiments of the invention.

It will be clearly understood that, although a number of prior art publications are referred to herein, this reference does not constitute an admission that any of these documents forms part of the common general knowledge in the art, in Australia or in any other country.

Claims (8)

1. A selective androgen receptor modulator compound having in-vivo androgenic and anabolic activity of a nonsteroidal ligand for the androgen receptor, the selective androgen receptor modulator compound represented by the structure of formula II below, Y NH X H 3 C "OH 0 wherein X is O, CH 2 NH, Se, PR or NR; SZ is a hydrogen bond acceptor, NO 2 CN, COR, CONHR; Y is a lipid soluble group, I,CF 3 Br, Cl, SnR 3 R is an alkyl or aryl group or OH; and Q is acetamido, trifluoracetamido, alkylamines, ether, alkyl, N-sulfonyl, O-sulfonyl, alkylsulfonyl, carbonyl or a ketone.

2. The selective androgen receptor modulator compound of claim 1, wherein X is O, Z is NO 2 Y is lipid soluble, and Q is acetamido.

3. Use of the compound of claim 1 or 2 for the preparation of a medicament for use in hormone therapy in a subject.

4. Use of the compound of claim 1 or 2 for the preparation of a medicament for use in treating a hormone-dependent condition in a subject.

Use of the compound of claim 1 or 2 for the preparation of a medicament for use in treating prostate cancer in a subject.

6. A method of hormone therapy comprising administering a compound as claimed in claim 1 or 2 to a subject.

7. A method of treating a hormone-dependent condition comprising administering a compound as claimed in claim 1 or 2 to a subject.

8. A method of treating prostrate cancer comprising administering a compound as claimed in claim 1 of 2 to a subject.