AU3405600A - Sex steroid activity inhibitors - Google Patents

Description

.60.

o

AUSTRALIA

Patents Act 1990 TRUE COPY COMPLETE SPECIFICATION STANDARD PATENT I certify that the following 213 pages are a true and correct copy of the description and claims of the original complete specification in respect of an invention entitled: SEX STEROID ACTIVITY INHIBITORS Applicant(s): ENDORECHERCHE, INC.

4 Signature: 1A- SEX STEROID ACTIVITY INHIBITORS .BACKGROUND OF THE INVENTION This invention relates to novel inhibitors of sex steroid activity such as antiestrogen compounds having effective antagonistic capability while substantially lacking agonistic effects. More particularly, certain preferred embodiments of the invention relate to certain estradiol and diphenylethylene analogs which have high affinity for estrogen receptors but do not activate such receptors and/or which inhibit the production of sex steroids or their precursors.

During the treatment of certain sex steroid-dependent diseases, it is important to greatly reduce or, if possible, eliminate certain sex steroid-induced effects. For this purpose, it is desirable both to block receptor sites stimulated by sex steroids and also to reduce the amount of sex steroid available to act at these sites. For example, alternative or concurrent therapy to administration of antiestrogens could involve attempts to block the production of estrogens by ovariectomy) such that less is available to activate receptor sites. However, prior art methods for blocking estrogen production insufficiently inhibit estrogen-induced functions. Indeed, it is possible that even in the total absence of sex steroid, some receptors may be activated. See Simard and Labrie, "Keoxifene shows pure antiestrogenic activity in pituitary gonadotrophs", Mol.

Cell. Endocrinol. 39: 141-144, (1985), especially page 144.

Hence, antagonists of sex steroids may produce greater therapeutic results than therapy which only inhibits sex steroid production. Prior art antagonists, however, often have insufficient affinity for receptors, and some, although capable of binding the receptors, may themselves act as agonists and undesirably activate the very receptors they are intended to shield from activation.

o There is, therefore, a need in the art for antiestrogens which effectively block estrogen receptors with minimal or no agonistic effect. In Wakeling and Bowler, "Steroidal Pure Antioestrogens", J. Endocrinol. 112: R7-R10 (1987), a steroid derivative is said to act as an antiestrogen but to exhibit some estrogen activity. The net effectiveness of a compound is effected by both its agonistic (undesirable) and antagonistic (desirable) activities.

In U.S. Patent 4,094,994, it is disclosed that the use of certain antiestrogens may inhibit certain human breast tumor cells.

H. Mouridsen et al., Cancer Treatm. Rev. 5: 131-141 (1978), discloses that Tamoxifen, an antiestrogen, is effective in remission of advanced breast cancer in about 30 percent of the women patients treated.

:The combined use of the antiestrogen Tamoxifen and a luteinizing hormone-releasing hormone agonist, Buserelin, is also known for treatment of breast cancer. See, for instance, Klijn et al. J. Steroid Biochem. 420: no. 6B, 1381 (1984). The objective remission of such cancers, however, remains unacceptably low.

It has been found that certain 7a-substituted derivatives of estradiol, for example a 7a-(CH 2 )10CONMeBu substitution possess antiestrogenic activity :(Bowler et al., 1985; Eur. Patent Application 0138504; Wakeling and Bowler, J.

Steroid Biochem. 30: 141-147 (1988). See also US patent 4,659,516. The substitution (CH 2 9 SOC5H 6

F

5 has also been used on certain compounds (Wakeling et al., Cancer Res. 51: 3867-3873, 1991).

Certain -(CH 2 10 CONMeBu substituted compounds are also disclused in US Patent 4,732,912 (See e.g. example 5 and 16). See also EP Pat No. 166 509, EP Pat No. 124 369, EP Pat. No. 160 508, EP Pat. No. 163 416, U.S. Pat No. 4,760,061, U.S.

Pat. No. 4,751,240 and Wakeling A.E. and Bowler, J. Endocrinol. 112: R7-R10 (1987).

Estradiol derivatives bearing a carboxyalkyl substituent at the 7a-position maintained their affinity for the estrogen receptor when linked via their carboxy group to agarose or polyacrylamide resin for affinity chromatography purification of the estrogen receptor (Bucourt et al., J. Biol. Chem. 253: 8221, S1978).

Some steroid derivatives, such as 16-methylene estradiol and 16-methylene estrone, have been described as inhibitors of 17p-hydroxysteroid dehydrogenase activity (Thomas et al., J. Biol. Chem. 258: 11500, 1983).

Certain nonsteroidal compounds which are stated to have antiandrogenic effect are described by Furr et al., J. Endocrinol. 113: R7-R9 (1987).

U.S. Pat. No. 4,659,695 relates to a method of treatment of prostate cancer for susceptible male animals including humans whose testicular hormonal secretions are blocked by surgical or chemical means, by use of an LHRH agonist, [D-Trp 6 des-Gly-NH21 0 ]LHRH ethylamide. The treatment includes administering an antiandrogen, flutamide in association with at least one inhibitor of sex steroid biosynthesis, aminoglutethimide and/or ketoconazole. See also PCT/U.S. 85/01454 (International Publication Number WO 86/01105) regarding combination therapy for treating hormonal-dependent cancers.

U.S. Pat. No. 4,472,382 relates to a method of treating prostate cancer using the combination of an antiandrogen and an LHRH agonist.

In U.S. Pat. No. 4,386,080 relates to new amide derivatives, and more particularly to novel acylanilides, possessing antiandrogenic properties.

In French Patent 2528434 and in Jordan and Koch, "Regulation of Prolactin Synthesis in vitro by estrogenic and antiestrogenic derivatives of estradiol and Estrone", Endocrinology 124(4): 1717-1725 (1989), antiestrogenic effects are described for certain 11p-substituted estradiol derivatives.

In U.S. Pat No. 3,995,060, U.S. Pat No. 4,161,540 and U.S. Pat. No. 4,139,638, it is disclosed that certain 4'-substituted and 3'-,4'-disubstituted anilides have antiandrogenic properties.

For a number of years, researchers have attempted to develop compounds which can efficiently inhibit androgen and/or estrogen formation without causing adverse effects to healthy tissues. More particularly, the inhibition of 17p-hydroxysteroid dehydrogenase, which is involved in the biosynthesis of testosterone, androst-5-ene-3p,17p-diol and estradiol, has been studied by some workers. Some affinity-label inhibitors for human placental estradiol 170-dehydrogenase have been described Chin and J.C. Warren, J. Biol.

Chem. 250: 7682-7686, 1975; Y.M. Bhatnagar et al., J. Biol. Chem. 253: 811-815, 1978; C.C. Chin et al., J. Biol. Chem. 255: 366G-3664, 1980; J.L. Thomas and R-C.

Strickler, J. Biol. Chem. 258: 1587-1590, 1983).

B. Tobias et al., J. Biol. Chem. 257: 27&3-2786 (1982) and R-J. Auchus and D.F.* Covey, Biochemistry 25: 7295-7300 (1986) disclose, respectively, the use of 7p-propynyl-substituted proges tins and propynyl-substituted 3-hydroxy-14,15-secoestra-1,3,5(10)-trien-17-one as inhibitors of the 17p-estradiol dehydrogenase.

Thomas J.L. et al., J. Biol. Chem. 258: 11500 (1983) have described that 16-methylene estradiol and 16-methylene estrone are inhibitors of 171 -hydroxysteroid dehydrogenase activity.

:Prior art methods have not been completely effective in inhibiting sex steroid synthesis while avoiding undesirable side effects.

Von Angerer et al. discuss other antiestrogens in "1-(aminoalkyl)-2phenylindoles as Novel Pure Estrogen Antagonists", J. Med. Chem. 1990; 33: 2635-2640. In U.S. Patent 4,094,994, where it is said that the use of certain antiestrogens; inhibit certain human breast tumor cells. See also DE 3821148.

A. Saeed et al., J. Med. Chem. 33: 3210-3216, 1990; A.P. Sharma et al., j Med.Chem. 33: 3216-3222 and 3222-3229 (1990) described the -synthesis and -7biological activities of 2,3-diaryl-2H-l-benzopyrans analogs as antiestrogens having the following molecular structure: a a* a.

a.

a.

a f ft ft N. Durani et al., J. Med. Chem. 32: 1700-1707 (1989) describe the synthesis and biological activities of benzofuran and triarylfuran analogues as antiestrogens.

The European counterpart of priority applications 07/377,010 and 07/265,150 was published on May 9, 1990 as European Application No 0367576. The European Search report for that case disclosed the following publications: In E.P. Patent No 305 242, Nique et al relates to the synthesis and the use of 17acyl steroids as drugs. The Search Report emphasized page 7, compound I'C.

In E.P. Patent No 280 618, Nique et al. relates to 7-substituted 19-nor-steroids for drugs. The Search Report emphasized Examples 2, 3, pages 22, 23 and the claims.

-8- In D.E. Patent No 32 42 894 Al, Neef et al relates to 17a-substituted equilenin for inhibition of progesterone biosynthesis and control of the fertility.

In U.S. Patent No 2,875,199, Cella, J.A. discuss 17-carboxylated estradiols for decreasing the serum concentration of cholesterol.

Blickenstaff et al. (Steroids, Vol. 46, No 4 et 5, pages 889-902) described the synthesis of 16 and 17-substituted estradiols suitable for coupling to vinblastine species.

Other Search Report References were previously discussed herein.

o* ot* ft OBJECTS OF THE INVENTION It is an object of the present invention to provide methods of inhibiting sex steroid activity. Such methods may be useful in the treatment of sex steroid-related diseases.

It is another object of the invention to provide a pure antiestrogen for therapeutic use.

It is another object of the invention to provide compositions capable of inhibiting sex steroid synthesis, especially estrogen synthesis.

It is another object to provide an antiestrogen having good affinity for estrogen receptors, but substantially lacking undesirable agonistic activity regarding these receptors and substantially lacking hormonal activity.

It is another object of the invention to provide a therapeutic antiestrogenic composition useful in the treatment of estrogen-related diseases. These diseases include, but are not limited to breast cancer, uterine cancer, ovarian cancer, endometriosis, uterine fibroma, precocious puberty and benign prostatic hyperplasia.

It is another object of the invention to provide inhibitors of sex steroid activity useful in the treatment of both estrogen- and androgen-related diseases.

Androgen-related diseases include but are not limited to prostate cancer, acne 10 vulgans, hirsutism, precocious puberty, benign prostatic hyperplasia, seborrhea, androgenic alopecia and sexual deviants. Control of androgen activity may also be useful in male contraception.

@6 4e U 6*S@ 6@ S 0 *4e 0 0 6**0 0*S@ 0S SO 0 S 0

S

0 4*O0 0e I, *46 U *05* @6 6@ 0@@SSS 6 -11- SUMMARY OF THE INVENTION In accordance with the invention, a pharmaceutical composition is provided comprising a therapeutically effective amount of the diphenylethylene derivatives specified herein. One embodiment of the diphenylethyl framework is illustrated below: R 5 R7 Re where the dotted line represents an optional double bond of Z or E configuration. Certain preferred substituents include, but are not limited to the following:

R

1 Rs, R8, and R 1 2 are preferably independently selected from the group consisting of hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy, alkylsulfonyl lower alkoxy, arylsulfonyl lower alkoxy, lower alkylsilyl, amino, nitro, nitrile and nitroso.

12-

R

2

R

4 R9 and Rll are preferably independently selected from the group consisting of hydrogen, hyd roxyl, halogen, lower alkyl, lower alkoxy, alkyisulfonyl lower alkoxy, arylsulfonyl -lower alkoxy, lower alkylsilyl, amino, nitrile, nitro, nitroso, azido, (Cl-C 7 alkanoyl mercuryl, lower alkylamino, dilower alkylainino, AXR 2 1

Y

7 -Al[Y-Al' 1

I-XR

2 j, and Al[Y-All]u-XR 2 1 wherein: A is straight- or branched-chain (Cl-C 3 0 alkylene, (C 2

-C

3 0) alkenylene, (C 2

-C

3 o) alkynylene or fluoro-substituted analogs of the foregoing; wherein u is an integer from 0 to 5; wherein Y7 is absent or selected from the group consisting of carbonyl and carboxyl, Al and All may be the same or different and are independently either absent or selected from the group consisting of straight- or branched-chain alkylene, straight- or branched-chain alkynylene, straight- or branched-chain alkenylene, phenylene and fluoro-substituted analogs of the foregoing, wherein Al and All together have a total of from 3 to 20 carbon atoms and Y is selected from the group consisting :of -SO 2

-NR

2 2 SiR 2 2

R

22 -CR22OR 2 2 -NR22CO-, -NR22CS-, -CONR22-, -COO-, -C OS-, -5CC-, -CSS-, -SCS-, -OCO- and phenylene (R 2 2 being hydrogen or lower alkyl), wherein

R

2 1 is selected from the group consisting of hydrogen, straight- or branched-chain lower alkyl, lower alkenyl or lower alkynyl, (C 3

-C

7 cycloalkyl, halogeno(lower)alkyl, carboxy (lower) al kyl, (lower)alkoxycarbonyl (lower)alkyl, (C 6 -Cl 0 aryl, (C 6

-C

1 o) arylalkyl, 13 di(lower)alkylamino(lower)alkyl and fluoro-substituted analogs of the foregoing, and wherein X is selected from the group consisting of

-CONR

23

-,-CSNR

2 3

-NR

2 4 CO-, -NR 2 4

CS-,-NR

2 4

CONR

2 3

-NR

24 -C(NR5)-NR23, -SO 2 NR23-, -CSS-, -SCS-, -NR 2 3 -(PO)R23-, -NR 2 4 COO-, -NR 2 4 SQ-, -SO- and -SO 2

(R

23 being selected from the group consisting of hydrogen, lower alkyl, a species which, together with R2 1 forms a saturated or unsaturated heterocyclic ring having at least one nitrogen atom and in certain embodiments, at least one other heteroatom selected from the group consisting of oxygen, sulfur, silicon, selenium and nitrogen, and fluoro-substituted analogs of the foregoing; and R 24 being hydrogen or lower alkyl, and R25 being hydrogen, nitrile or nitro). In certain preferred enbodiments, XR21 forms a tetrazole ring, CON CxHx CSN.C.H2x or -N C.HH (where x is an integer from 4-6).

R

3 and RIO are preferably independently selected from the group consisting of hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy, lower alkoxy ~~carbonyloxy, carboxyl, (Cl-C 2 0 alikanoyloxy, (C 3

-C

2 o) alkenoyloxy, 3 Co alkynoyloxy, (C7-CU1) aroyloxy and alkylsilyloxy.

R

6 and R7 are preferably independently selected from the group consisting of hydrogen, amino, lower alkylamino, dilower alkyl amino, nitro, nitrile, nitroso, halogen, lower alkyl, lower alkenyl, lower alkynyl, halogeno lower alkyl, halogeno lower alkenyl, halogeno lower alkynyl, alkyl sulfonyl, aryl 14sulfonyl, a substituted 5 to 7 member heterocyclic. ring having at least one heteroatom (selected from oxygen, sulfur, silicon, selenium, nitrogen), -CH2)sW (wherein- W is nitrile, hydroxyl, azido, nitroso, alkoxy, nitro, thionitrile, halogen, alkyl sulfonyl or aryl sulfonyl and s is an integer from 1 to a moiety of the formula: wherein: F is absent or selected from the group consisting of alkyl, carbonyl or carboxyl, wherein the phenyl ring may be halogenated, wherein R61 is hydrogen, hydroxyl, halogen, lower alkyl, lower alkenyl, lower :alkynyl, nitrile, nitro, nitroso or X 6

(CH

2 )ny 6

(X

6 being selected from the group consisting of Se-, -SC) 2 and and Y 6 :being selected from the group consisting of hydroxyl, amino, inonoalkyl amino, dialkyl amidno, dimethyl N-oxide, N-aziridyl, guanidine, N-pyrrolidino, N-piperidino, N-methylpiperazino, N-morpholino and alkoxy, and n being an integer from 1 to 6, preferably 3),

AXCR

2 j, Y7-A-[Y-A11]uXR 2 1, and AI-[Y-A 1 1]u-XR 2 1, wherein: A is selected from the group consisting of straight- or branched-chain (CI -C 30 alkylene, (C 2

-C

30 alkenylene, (C 2

-C

30 alkynylene and fluoro-substituted analogs of the foregoing, wherein u is an integer from 0 to 5, wherein Y 7 is absent or is selected from the group consisting of carbonyl, carboxyl, -CH 2 S- and -0H 2 wherein Al and All may be the same or different and may be absent or selected from the group consisting of straight- or branched-chain alkylene, straightbranched-chain alkynylene, straight- or branched chain :alkenylene, phenylene and fluoro-substituted analogs of the foregoing, wherein Al and All together have a total of from 2 to carbon atoms, wherein Y is selected from the group consisting of -NR22CS-, -CONR22-, -CSNR 22 -COG-, -COS-I, -CSS-, -SCS-, -0CC)- and phenylene (R2 2 being hydrogen or lower alkyl), wherein

:R

21 is selected from the group consisting of hydrogen, straight or branched chain lower alkyl, lower alkenyl, lower alkynyl, (C 3

-C

7 cycloalkyl, halogeno (lower) alkyl, carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower) alkyl, (C6-Clo)aryl, (C 7

-C

11 arylalkyl, di(lower) alkylamino (lower) alkyl and fluoro-substituted analogs of the foregoing, wherein X is selected from the group consisting of

-CC)NR

2 3

-CSNR

23

-NR

24 CO-, -NR 24 CS-, -NR 2 4

CONR

2 3

-NR

24

-C(NR

25

)-NR

23 -S0 2

NR

23 -CSS-, -SCS-, -NR 23

-(NO)R

23

-(PO)R

23

-NR

24 COO-, -NR 24 S0 2 -SO- and -S02- (R 23 -16being selected from the group consisting of hydrogen, lower alkyl and a species which, together with R 2 1 forms a saturated or unsaturated heterocyclic ring having at least one nitrogen atom and, in certain embodiments at least one other heteroatom selected from the group consisting of oxygen, sulfur, silicon, selenium and nitrogen, and fluoro-substituted analogs of the foregoing, R 24 being hydrogen or lower alkyl and R 25 being hydrogen, nitrile or nitro). In certain S. preferred embodiments, XR 21 forms a tetrazole ring.

R

6 and R 7 may also be a species which, in combination with another substituent of general molecular formula I, forms a moiety selected from the group consisting of: -CH 2 -CHX-, -CX 2 (X being halogen, carboxyl or alkoxycarbonyl), >N-CN, >NR 2 9 and >NC0 2

R

2 9-(R 2 9 being hydroxy or lower alkyl), lower alkylene, -(CH 2 )rO(CH 2 -(CH2)rS(CH2)s-, -(CH2)rSe(CH 2 s -(CH2)rSO(CH2)s-, -(CH2)rSO2(CH2)s-, (CH 2 )rCO(CH 2 -(CH2)r(CH2)NR CH2)s-, -(CH2)rSiR22R22(CH 2 or -(CH2)rCR22(CH2)s- (R22 being hydrogen or lower alkyl, r and s being independent integers from 0 to a moiety of the formula: R71 i.rA'-[Y-A 1 ],XR2, I

I

-17wherein

A

1 Y, A 11 u, X and R 2 1 are as defined above, wherein Z is absent or is selected from the group consisting of lower alkylene, halogeno lower alkylene,-(CH2)nO-, -(CH2)nS-, -(CH2)nSe-, -(CH2)nSO-, 2)nSO2-, -(CH2)nCO-, -(CH2)nNR22-, -(CH2)nSiR22R22- and -(CH2)nCR22OR22-, R12 is as defined above, n being an integer from 0 to 3, and R71 being selected from a group consisting of *hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy and lower alkylsilyl, a moiety of the formula: >N-AI[Y,-All]u XR 21

IV

wherein N is nitrogen atom and A 1 Y, A 11 u, X and R 2 1 are as defined above.

V

In preferred embodiments, moieties which are combinations of R groups from general structure I, are combinations of R 6 and R7, R 6 with R 1 or R 12 or R 7 with or Rg.

The invention further provides an inhibitor of sex steroid activity having, as part of its molecular structure, a substituted or unsubstituted estrogenic nucleus of general formula V: -18- 11 ORe 1 7 4 5 eRs

V

wherein R 5 and R 6 are hydrogen lower alkyl, alkoxy carbonyl, (CI-C 20 alkanoyl,

(C

3 -C20) alkenoyl, (C 3

-C

20 alkynoyl, (C7-C11) aroyl and alkylsilyl, wherein dotted lines are optional pi bonds. In some embodiments, the optional pi bonds are not simultaneously present when aromaticity would result from such simultaneous presence; R 15 is either a direct bond from e to the number carbon or a methylene or ethylene linkage to the number 5 carbon or a lower alkyl substituent, wherein e is selected from the group consisting of carbon, sulfur and nitrogen, q is absent or is a divalent methyl or ethyl moiety; said inhibitor further having a side chain of the formula -RI[-B-R 2 x L-G wherein in at least one of said side chains is substituted at a position selected from the group consisting of carbon 2, carbon 4, carbon 5, carbon 10, carbon 11, carbon 13, q and e atom wherein: -19x is an integer from 0 to 6, wherein at least one of L and G is a polar moiety distanced from the substitution point by at least three intervening atoms, and wherein:

R

1 and R 2 are independently either absent or selected from the group consisting of straight- or branched-chain alkylene, straight- or branched-chain alkynylene, straight- or branched-chain alkenylene, phenylene, and fluoro-substituted *e*o analogs of the foregoing; B is either absent or selected from the group consisting of -SO-, -SO2-, -NR 3 -SiR 3 2

-CR

3

OR

3

-NR

3 CO-, -NR 3 CS-, -CONR 3

-CSNR

3

-COO-,

-COS-, -SCO-, -CSS-, -SCS-, -OCO- and phenylene (R 3 being hydrogen or lower alkyl); L is either a moiety which together with G, forms a heterocyclic ring having at least one nitrogen atom or is selected from the group consisting of lower alkyl,

-CONR

4

-CSNR

4

-NR

5 CO-, -NR 5 CS-, -NR 5

CONR

4

-NRSC(NR

6

)-NR

4 -S02NR4-, -CSS-, -SCS-, -NR5COO-, -NRSS02-, -NR4-, -SO- and -SO 2

(R

4 and R 5 being independently selected from the group consisting of hydrogen and lower alkyl; and R 6 being selected from the group consisting of hydrogen, nitrile and nitro); and G is either a moiety which together with L forms a heterocyclic ring having at least one nitrogen atom or is selected from the group consisting of hydrogen, lower alkyl, lower alkenyl, lower alkynyl, (C 3

-C

7 cydoalkyl, bromo(lower)alkyl, chloro(lower)alkyl, fluoro(lower)alkyl, iodo(lower)alkyl, cyano(lower)alkyl, carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower)alkyl, (C6-Ci0) aryl, (C 7 Cll)arylalkyl, di(lower)alkylamino(lower)alkyl, fluoro-substituted analogs of the foregoing.

This invention further provides a pharmaceutical composition comprising a S: pharmaceutically acceptable diluent or carrier and a therapeutically effective amount of the foregoing sex steroid activity inhibitor.

The inhibitor is preferably hydroxy-substituted in at least the 3 or 12 positions, and is preferably substituted at the 7 position with a Ci-C 4 alkyl. Compounds of formula V above may be used, preferably as part of pharmaceutical compositions including acceptable diluents or carriers, to treat sex steroid dependent diseases by inhibiting sex steroid activity.

In another embodiment, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a therapeutically effective amount of a sex steroid activity inhibitor having, as part of its molecular structure, an estrogenic nucleus of the formula: -21 wherein said sex steroid activity inhibitor includes, as another part of its molecular structure, a side chain of the formula RI(B-R 2 )xLG substituted onto a ring carbon of said estrogenic nucleus to form: R6 I Ir I I" R3. Z R'(B-R 2

LG

wherein x is an integer from 0 to 6, wherein at least one of L and C is a polar moiety distanced from said ring carbon by at least three intervening atoms, and wherein:

R

1 and R 2 are independently either absent or selected from the group consisting of straight- or branched-chain alkylene, straight- or branched-chain alkynylene, straight- or branched-chain alkenylene, phenylene, and fluoro-substituted analogs of the foregoing; B is either absent or selected from the group consisting of -SO-,

-SO

2

-NR

3 -SiR 30

-CR

30

OR

30

-NR

3 CO-, -NR 3 0 CS-, -CONR 3

-CSNR

30 -COO-, -COS-, -SCO-, -CSS-, -SCS-, -OCO- and phenylene (R 30 being hydrogen or lower alkyl); -22 L is either a moiety which together with G, forms a heterocyclic ring having at least one nitrogen atom or is selected from the group consisting of lower alkyl,

-CONR

4

-CSNR

4

-NR

5 CO-, -NR 5 CS-, -NR 5

CONR

4

-NR

5

C(NR

6 0)NR 4

-SO

2

NR

4 -CSS-, -SCS-, -(NO)R 4

-(PO)R

4

-NR

5 COO-, -NR 5 COO-, -NR 5

SO

2

-NR

4 and -SO 2 (R4 and R 5 being independentl y selected from the group consisting of hydrogen and lower alkyl; and R 60 being selected from the group consisting of hydrogen, nitrile and nitro); G is either a moiety whichi together with L forms a heterocyclic ring having at least one nitrogen atom or is selected from the group consisting of hydrogen, lower alkyl, lower alkenyl, lower alkynyl, (C3-C7)cydloalkyl, bromo(lower)alkyl, chloro(lower)alkyl, fluoro(lower)alkyl, iodo(lower)alkyl, cyano(lower)alkyl, carboxy(lower)alkyl, (lower)alkoxycarbonyl~lower) alkyl, (C 6 -C iO)aryl, (C 7

C

11 )arylalkyl, di(lower)alkylamidno(lower)alkyl, and fluoro-substituted analogs of the foregoing; wherein Z is selected from the group consisting of lower alkylene, halogeno lower alkylene, -(CH2)nO-, -(CH 2 )nrS-, -(CH 2 )nSe-, -(CH 2 )nSO-, -(CH 2 nSO2-, -(CH2)nCO-, -(CH 2 )nNR22-, -(CH2)nSiR2R~r and -(CH 2 )nCR2OR2 (wherein R22 is hydrogen or lower alkyl and n is an integer from 0 to 3); wherein R 3 and R 10 are independently selected from the group consisting of hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy, lo*er alkoxy carbonyloxy, carboxyl, (C 1

-C

2 o) alkanoyloxy, (C 3

-C

20 alkenoyloxy; (C 3

-C

20 -23alkynoyloxy, (C7-C) aroyloxy alkylsilyloxy; OR' 3 or OR' 10 (whererin R' 3 and

R'

10 are hydrogen, alkyl, (Ci-C 20 alkanoyl, (C 3

-C

20 alkenoyl, (C3-C 20 alkynoyl or (C7-C 11 aroyl); and wherein R 6 is selected from the group consisting of hydrogen, lower alkyl, lower alkenyl and lower alkynyl.

when L and G are together a nitrogen hetero ring, such ring is preferably -N^CxH 2 x or -N^Cx-1H 2 x-20 (where x is an integer from 4-6).

R6 is preferably methyl, ethyl or propyl. Unsaturated analogs such as ethenyl or ethynyl may also be used. In some embodiments, at least one of the A and D rings is substituted with hydroxyl or a moiety converted in vivo to hydroxyl hydrogen in positions 3 and 10, acetoxy, benzoyloxy, akanoyloxy, alkenoyloxy and aroyloxy). For example, R 3 and/or R 10 may be hydroxyl. The ring-closing moiety, Z, is preferably -NH- or -CH 2 and in some embodiments is a bivalent moiety containing carbon, sulfur or nitrogen.

Preferred side chains RI(B-R 2 )xLG) are discussed in the detailed description and examples herein.

-24- In another embodiment, the estrogenic nucleus may include a ring nitrogen onto which is substituted the side chain. Thus, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and a therapeutically effective amount of a compound of the formula: R6 Rs1 A B N G' wherein the dotted line represents an optional double bond, wherein R 5 and R 6 are independently hydrogen, hydroxy or a moiety which is converted to hydroxy in vivo; wherein R 100 is bivalent moiety which distances L from the B-ring by 4-10 Sintervening atoms; wherein L is a bivalent or trivalent polar moiety selected from the group consisting of -CON<, and -SON<; wherein G I is either absent or selected from the group consisting of hydrogen, a C 1 to C 5 hydrocarbon, a substituted or unsubstituted C 5 to C 7 cycloalkyl, a bivalent moiety which joins G 2 and L to form a 5- to 7-membered heterocyclic ring, and halo-substituted derivatives of the foregoing; and wherein G 2 is either absent or selected from the group consisting of hydrogen, a C1 to C 5 hydrocarbon, a substituted or unsubstituted bivalent moiety which joins G 1 and L to form a 5- to 7-membered heterocyclic ring and halo-substituted derivatives of the foregoing.

In another embodiment, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and a therapeutically effective amount of an estrogen activity inhibitor of the following formula: o 11 *R6 4 G A B 7 14 Rs R G1

G

2 wherein the dotted line is an optional double bond; wherein R 5 and R 6 are independently hydrogen, hydroxyl or a moiety which is converted to hydroxyl in vivo; wherein Z is a bivalent ring closing moiety.

wherein R 10 0 is a bivalent moiety which distances L from the B-ring by 4-10 intervening atoms; -26wherein L is a bivalent or trivalent polar moiety selected from the group consisting of -CON<, and -SON<.

wherein G 1 is either absent or selected from the group consisting of hydrogen, a

C

1 to C 5 hydrocarbon, a saturated or unsaturated C 5 to C7 cycloalkyl, a bivalent moiety which joins G 2 and L to form a 5- to 7-membered heterocyclic ring, and halo-substituted derivatives of the foregoing; a i.:":wherein G 2 is either absent or selected from the group consisting of hydrogen; a a

C

1 to C 5 hydrocarbon, a substituted or unsubstituted C5 to C 7 cycloalkyl, a bivalent moiety which joins G 1 and L to form a 5- to 7-membered. heterocyclic ring and halo-substituted derivatives of the foregoing.

*0o0 oo..

In one embodiment, Z includes a carbon, sulfur or nitrogen atom. In another

G'.

embodiment, Z in -CH 2 or In the side chain R 1 0 0 it s1 preferred that at least one of G 1 and G 2 has at least two carbon atoms. G 1 and

G

2 in some embodiments, are independently absent or are selected from the group consisting of hydrogen, C 1 to C 5 hydrocarbon, a substituted or unsubstituted C 5 to C7 cycloalkyl and halo-substituted derivatives of the foregoing. R 100 in some embodiments, is a straight or branched chain alkylene, alkenylene or alkynylene which distances L from the B ring by 4-10 intervening atoms. Unsaturation in R 10 0 may include, for example, phenylene or alkynylene. Preferred moieties for R 100 include but are not limited to Rl(B-R 2 )x from the side chain RI(B-R 2 )xLG discussed above or from the side -27chain A'-(Y-A")uXR 2 1 discussed above. Preferred side chains include but are not limited to -(CH 2 1 oCONCH 3

C

4

H

9

-(CH

2 9 SOCsH6F 5

-(CH

2 6

NC

5

H

1 0 and -0-O(CH 2 2

NC

5

H

10 When G 1

G

2 and L combine to form a nitrogen-containing heterocyclic moiety, such moiety is preferably, but is not limited to, -N CxH2x or -N^Cx-iH2x-20 (where x is an integer from 5-7).

As used herein, the term "sex steroid activity inhibitor" includes any compound which suppresses the activity of sex steroids by any mechanism including, for example, inhibition of sex steroid synthesis or antagonistic blocking of sex steroid receptors. "Androgen activity inhibitors" and "estrogen activity inhibitors" are sex steroid inhibitors capable of inhibiting the activity of androgens and estrogens, respectively. For example, estrogen activity inhibitors include, but are not limited to antiestrogens which block estrogen receptors, thereby making them unavailable to estrogen compounds which could otherwise activate those receptors. Sex steroid activity inhibitors also include compounds which inhibit the formation of compounds capable of activating sex steroid receptors such as inhibitors of the production of natural sex steroids 17p-estradiol) or inhibitors of production of precursors of natural sex steroids. One mechanism by which these sex steroid production inhibitors may operate is by blocking enzymes which catalyze production of natural sex steroids or their precursors inhibitors of enzymes such as aromatase, 17p-hydroxysteroid dehydrogenase, 30-hydroxysteroid dehydrogenase and the like).

28- As used herein, the term "estrogenic nucleus" includes any compound which, in the absence of the side chain substituent specified herein, is capable of acting as an estrogen as determined by a weight increase of at least 100 percent over a seven-day period of the uterus of ovariectomized rats treated with the compound in question (0.5 mg twice daily per 100 grams of body weight) versus a control group of ovariectomized rats. Treatment should start on the day of castration. The precise test, other than any parameters set forth in this paragraph, is that reported in Simard et al., Mol. Endocrinol. 2: 775-784 (1988).

sees oo0o The following conventions apply to structural formulae set forth herein.

Unless specifically designated to the contrary, substituents may have either a or p stereochemistry or, where valence permits may represent one substituent in. a

*OSS

S

position and another in p position. Presence of optional pi bonds are independent of each other. All structures include salts thereof. Atoms of any sees estrogenic nucleus for which no substituent is shown or described may optionally be substituted or unsubstituted so long as such substitution does not prevent the nucleus from functioning as an "estrogenic nucleus" as defined herein. Those atoms having a defined substituent may optionally be further substituted by other substituents where their valence permits such further substitution. As used herein, the term "lower", when describing a chemical moiety means a moiety having 8 or fewer atoms. For instance, a "lower alkyl" means a C 1 to C 8 alkyl. Any moiety of more than two atoms may be straight- or branched-chain unless otherwise specified.

-29- BRIEF DESCRIFFION OF DRAWINGS Figure 1 illustrates a competition binding assay of the affinity of estradiol, diethylstilbestrol, ICI 164384 (Wakeling, A.E .and Bowler, 1987; J. Endocrinol.

112: R7-R110) and EM-142 (an antiestrogen having a nonsteroidal nucleus and synthesized in example 1, herein) for the rat uterine cytosol receptor (Asselin et al., 1978; J. Steroid Biochem. 9: 1079-1082).

Figure 2 illustrates the effect of the indicated doses of EM-142 injected twice daily on uterine weight (mg) in adult female ovariectomized Balb/C mice treated for 4.5 days in the presence or absence of simultaneous treatment with 17p-estradiol (0.01 jg, Figure 3 illustrates the effect of the administration of EM 139 on uterine weight (mg) in adult ovariectomized mice. The compound was administered twice daily at the indicated doses for 4.5 days in the presence or absence of 17festradiol (0.01 pg, via the subcutaneous route.

Figure 4 is a graph illustrating that the antiestrogen which is the subject of Figure 3 is also a good inhibitor of sex steroid synthesis.

Figure 5 illustrates the comparative inhibitory activity of increasing concentration of EM 343 and EM 312 on the growth of human breast cancer ZR-75-1 cells stimulated by 17p-estradiol. The respective IC50 values are calculated at 2.55 x 10- 1 0 M for EM 343 and 8.43x10- 1 0 M for EM 312, thus indicating a 3-fold higher activity for EM 343.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS In certain preferred embodiments of the invention, the R 3 and R 10 substituents on the nucleus of structure 1 supra are hydroxyl, (C 1

-C

2 0) alkanoyloxy (C 3

-C

2 0) alkenoyloxy, (C 3 -C20) alkynoyloxy, (C7-C10) aroyloxy and/or the R7 substituent is Al-[Y-A 1 1 lu-X-R 21 It is also preferred that the R7 substituent have between 7 and 20 carbon atoms. It is also preferred that R 6 of structure I, supra be lower alkyl, ethyl, fluoroethyl, or (CH2)2W, wherein W is a halogen or lower alkoxy, unsaturated lower alkenyl or alkynyl groups may also be used. In certain embodiments, therapeutic compositions may be comprise one or more compounds represented by Formula L Preferably, at least one antiestrogenic compound is represented by the formula: 6 R11 0 o

R

2 1 X-[A11-Y]u-A 1 R9 VI wherein A 1 All, Y, u, X and R 2 1 are defined as previously for R 6 and R 7 in the formula I, wherein the double bond is in trans configuration, -31wherein R' 3 and R 10 o are hydrogen, alkyl, (Cl-C 2 o) alkanoyl (C 3

-C

20 alkenoyl, (C3-C2o) alkynoyl or (C7-CllOaroyl, wherein R 6 is preferably selected from the g-roup consisting of hydrogen, nidtro, nitrile, halogen, lower alkyl, lower alkynyl, halogeno lower alkyl, halogeno lower alkenyl, halogeno lower alkynyl, alkyl sulfonyl, aryl sulfonyl, a :::.substituted 5 to 7 member heterocyclic ring having at least one hetero atom :(selected from oxygen, sulfur, silicon, selenium, nitrogen), -(CH 2 )SW (wherein W is nitrile, hydroxyl, azido, nitroso, alkoxy, nitro, thionitrile, halogen, alkyl sulfonyl, aryl sulfonyl and s is an integer from 1 to or has the formula: ri 61 :wherein: F is present or selected from the group consisting of alkyl, carbonyl or carboxyl, wherein the phenyl ring may be halogenated, wherein R61 is hydrogen, hydroxyl, halogen, lower alkyl, lower alkenyl, lower alkynyl, nitrile, nitro, nitroso or X6(CH2)ny 6 wherein X6 is selected from the group consisting -S02- and -CCO- and Y6 is selected from the group consisting hydroxyl, amino, monoalkyl -32amino, dialkyl amino, dimethyl N-oxide, N-aziridyl, guanidino, N-pyrrolidino, N-piperidino, N-methylpiperazino, N-morpholino and alkoxy, and n is an integer from 1 to 6 preferable 3.

R

9 and R 1 1 are preferably independently selected from the group consisting of hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy, lower alkylsilyl, amino, nitrile, nitro, nitroso, azido, lower alkylamino, dilower alkylamino, AXR 2 1 and Al[Y-All]u-X-R 2 1 wherein A, A 1

A

11 Y, X, R 2 1 and u are as defined previously from R 6 and R 7 When administered systemically, pharmaceuticals of the inventions may be used in the treatment of breast cancer, uterine cancer, ovarian cancer, endometriosis, uterine fibroma, precocious puberty and benign prostatic hyperplasia.

When sex steroid activity inhibitors are administered in accordance with the invention,they are preferably administered at a dosage from about 1 mg to about 2000 mg of active expedient sex steroid activity inhibitor), per day per kg of body weight, most preferably from about 10 mg to about 100 mg per day per 50 kg of body weight Pharmaceutical compositions comprise therapeutically effective amounts of one or more of the sex steroid activity inhibitors (including antiestrogens) discussed herein wherein a pharmaceutically acceptable diluent or carrier is -33included with the active compound(s). The diluent or carrier will vary in accordance with known techniques depending upon the manner in which the pharmaceutical composition is to be administered.

A composition suitable for oral administration may preferably include at least one inhibitor of sex steroid activity wherein the total concentration of all such inhibitors in said pharmaceutical composition is from about 1% to about 95% of the composition (by weight), and preferably -from about 5% to about 20%. The composition preferably further includes a pharmaceutically acceptable diluent, for example, starch or lactose with or without tartrazine.

When prepared for parenteral injection, an inhibitor of sex steroid activity is preferably added at a concentration between about 1 mg/ml and about 100 o mg/ml (preferably about 2 mg/ml to about 10 mg/ml) into a carrier preferably selected from the group consisting of saline, water, aqueous ethanol, aqueous dimethylsulfoxide and oil.

A composition suitable for parenteral administration preferably contains a carrier and an antiestrogen in accordance with the invention at a concentration sufficient to introduce from about 1 mg to about 1000 (preferably 5 to 50) mg of the antiestrogen per 50 kg of body weight per day. The volume flow will, of course, vary with the concentration at which the pharmaceutical composition is being administered.

-34- During the early course of treatment, it is preferred to take occasional blood samples and to alter dosage as necessary to maintain serum concentration of the sum of the active compounds between about 0.2 jig/ml and 10 gg/ml.

In certain alternative embodiments, the pharmaceutical composition of the invention may be formulated for sustained release in accordance with known techniques. These sustained release formulations are preferably prepared in an appropriate manner for either oral, intramuscular, or subcutaneous administration.

o* e Other alternative preferred embodiments include pharmaceutical compositions comprising therapeutically effective amounts of compounds of the formula:

S

s

OR'

10 o

R'

3 0 O R 10

R

21 X-[A11-Y1u-A1 VII

CC*

wherein the dotted line represents an optional pi bond, wherein R'3, R'IO, A 1 Y, A 1 1 X, -R 2 1 and u are defined as previously in formula VI especially A-[Y-All]u-XR21 is -CO-p O4-O-(CH 2 )nCONR2R22; wherein R 21 and R22 are defined as previously for R, 6 or R7 in formula I and n is an integer from 1 to 15; or.

R

2

,X(CH

2 )nO VinI wherein the dotted line represents optional double bond, especially in trans configuration, wherein R 3 R:1O, R6 are defined as previously, wherein R 2 1 is selected from the group consisting of hydlrogen, straight- or branched-chain lower alkyl, lower alkenyl or lower alkynyl, (C 3

-C

7 cycloalkyl, :halogeno~lower)alkyl, carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower)alkyl, (C 6-Ci o) aryl, (C 7

-C

1 1 )arylalkyl, di(lower)alkylamino (lower)alkyl and fluoro-substituted analogs of the foregoing, wherein X is -CONR -CNR2-

-NI(

24 C0-, -NR 2 4

-NR

2 4 CONR23-, -NR 2 4 C(NR2.)NR23-, -SO 2 NR23-, -GO-, -CSS-, -SCS-, -NR23-, -(NO)R23-, {PO)R23-, -NR 2 4 C0O-, -NR 2 4 S0 2

-SO-

or -SO 2 wherein R23 is selected from the group consisting of hydrogen, lower alkyl and a species which, together with R 2 1 forms a saturated or unsaturated heterocyclic ring having at least one nitrogen atom and fluoro-substituted analogs of the foregoing, wherein R 2 4 is hydrogen or lower alkyl and wherein R2 is hydrogen, nitiile or nitro; or XR 21 forms a tetrazole ring; or- -36-

R

6

N,

R-

3 0 I 1 Y']R1 wherein the dotted line represents an optional pi band, wherein W 3 13 6

RIO,

Al, All, Y, X, R(21 and u are as defined previously; or R6 1

-EY-A'

1 ]uXR 21 x -37 wherein the dotted line represents an optional pi bond, especially in trans configuration, wherein R' 3 RIO, R.

6 Al, All, Y, X, R 2 1 and u are defined as previously, wherein R 7 is preferably selected from the group consisting of hydrogen, halogen, lower ailkyl, amnino, nitro, nitroso, nitrile, lower alkylamidno and dilower alkylamino; or 2113 A B 7.1 7 A* v R-3 4 whri h ote ierpeensa pinl obebnwhri 3 !0 wherein~ AlA 1 AluX-[iYreere-aonfigrto n heenZi absent or selected from the group consisting of lower alkylene, halogeno lower alkylene, -(CH2)n0', -(CH2)nS-, -(CH2)nSe-,-(CH2)nSO-, -(CH2)nSO2-i -(CH2)nCO-, -(CH2)nNR2.2, -(CH2)nSiR22R22- or -(CH2)nCR22OR22-, (wherein R2 2 is defined as previously and n is an integer from 0 to 3).

Preferred methods of treating of sex steroid-related diseases, especially estrogen-related diseases, and preferred methods of blocking estrogen receptors comprise administering to a patient in need of such treatment, a therapeutically -38effective amount (discussed supra) of a sex steroid-activity inhibitor as defined above.

Preferred estrogenic nuclei suitable for substitution with the -RI[-B-R 2 ]xL-G side chain in accordance with the invention include but are not limited to compounds reported in the literature as having estrogenic activity, natural estrogens such as estradiol, estrogenic derivatives thereof, and other nuclei which provide the threshold increase in uterine weight of ovariectomized rats set forth above as defining an estrogenic nucleus (Simard et al., Mol.

EndocrinoL 2: 775-784, 1988).

Some preferred estrogenic nuclei include but are not limited to: OR4 wherein x is a halogen, preferably chlorine or iodine; -39wherein R 3 and R4 are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (CI-C 2 0 alkanoyl, (C 3

-C

20 alkenoyl, (C 3

-C

2 0 alkynoyl and (C 7

-C

1 1 aroyl, alkylsilyl, I-alkyloxy-alkyl and 1-alkyloxy cycloalkyl; or

OH

9.

9 99. 9 9999 .9 99 9 9 .99.

9 99*9 9 9 9*9 9 .9 9 99 9

R

3 0 wherein R 3 is selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (CI-C 2 0 alkanoyl, (C 3

-C

2 0 alkenoyl,

(C

3

-C

2 0 alkynoyl and (C 7

-C

1 1 aroyl, alkylsilyl, 1-alkyloxy-alkyl and I -alkyloxy cycloalkyl; or or

R

3 0 wherein R 3 is selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C 1

-C

2 0 alkanoyl, (C 3

-C

20 alkenoyl,

(C

3

-C

20 alkynoyl and (C 7

-C

1 1 aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cycloalkyl; or 9 9 9 9 *9*9 *9*9 *99* 9**9 9 9 *9*9 9 *0 9 9.9.

9 9. 9 .9 *99*9* 9 wherein R,3 is selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (Cl-C 2 0 alkanoyl, (C3-C2O) alkenoyl,

(C

3

-C

2 0 alkynoyl and (C7-CUI) aroyl, alkylsilyl, l-alkyloxy-alkyl and 1-alkyloxy cycloalkyl; or wherein R 3 is selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C 1

-C

2 alkanoyl, (C 3

-C

20 alkenoyl, -41-

(C

3

-C

2 0 alkynoyl and (C 7 -CII) aroyl, alkylsilyl, I-alkyloxy-alkyl and I -alkyloxy cydloalkyl; or 99 4* 9 99 S 9 9 9 *9*9 9*9* *9 99 9 0 wherein the dotted line are an optional double bonds; wherein R5 and R 6 are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (Cl-C 2 o), ailkanoyl, (C 3 -C20) alkenoyl, (C 3

-C

20 alkenoyl, (C 3

-C

2 o) alkynoyl. and

(C

7

-C

1 1 aroyl, alkylsilyl, I-alkyloxy-alkyl and 1-alkyloxy cydloalkyl; orfee 9 .9 9 000 9 09 wherein the dotted line is an optional double band; -42 wherein R.

5 and R.

6 are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (Cl-C 2 0 alkanoyl, (C 3

-C

2 o) alkenoyl, (C 3

-C

2 o) alkynoyl and (C 7

-C

1 1) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cycloalkyl; or R*J S.

0O R6 0 -43wherein the dotted line is an optional double bond wherein R 5 and R6 are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (Cl-C 20 alkanoyl, (C 3

-C

2 o) alkenoyl, (C 3

-C

20 alkynoyl and (C 7

-C

11 aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydloalkyl; or

S

S.

S S. wherein q is absent, methylene or ethylene wherein R 5 and R 6 are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C7'-C 11 aroyl, alkylsilyl, 1-alkyloxy-alkyl and I-alkyloxy cydloalkyl; -or- -44wherein Z is selected from the group consisting of lower alkylene, halogeno lower alkylene, -(CH2)nO-, -(CH1 2

-(CH

2 )nSe-, -(CH 2 )nSO-,

-(CH

2 )nSO27, -(CH2)CO, -(CH2)nNR22r, -(CH2),SiR22R22- or -(CH 2 )nCR22OR22 (wherein R22 is defined as previously and n is an integer from 0 to 3); wherein R3 and Rio are preferably independently selected from the group consisting of hydrogen, hydroxyl,

OR

3 1 ORIO', halogen, lower alkyl, lower alkoxy, lower alkoxy carbonyloxy, carboxyl, (Cl-C 20 alkanoyloxy,

(C

3

-C

2 0 alkenoyloxy (C3-CQo) alkynoyloxy, (C7-Cil) aroyloxy and alkylsilyloxy; wherein R' 3 and R'jo are hydrogen, alkyl, (Cl-C 2 0 alkanoyl, (C3-C2cj) alkenoyl, (C 3

-C

2 o) alkynoyl or (C 7 -CII) aroyl; wherein* R,6 is preferably selected from the group consisting of hydrogen, nitro, nitrile, halogen, lower alkyl, lower alkenyl, lower alkynyl, halogeno lower alkyl, halogeno lower alkenyl, halo~geno lower alkynyl, alkyl sulfonyl, aryl sulfonyl, a substituted 5 to 7 member heterocyclic ring having at least one hetero atom (selected from oxygen, sulfur, silicon, selenium, nitrogen),

-(CH

2 )sW (wherein W is nitrile, hydroxyl, azido, nitroso, alkoxy, nitro, thionitrile, halogen, alkyl sulfonyl, aryl sulfonyl and s in an integer from I to or has the formula: F Q 161 wherein: F is absent or selected from the group consisting of alkyl, carbonyl or carboxyl, wherein the phenyl ring may be halogenated, wherein R i is hydrogen, hydroxyl, halogen, lower alkyl, lower alkenyl, lower alkynyl, nitrile, nitro, nitroso or

X

6

(CH

2 )nY 6 wherein X6. is selected from the group consisting

-SO

2 and and Y 6 is selected from the group consisting hydroxyl, amino, monoalkyl amino, dialkyl amino, dimethyl N-oxide, N-aziridyl, guanidino, N-pyrrolidino, N-piperidino, N-methylpiperazino, N-morpholino and alkoxy, and n is an integer from 1 to 6 preferably 3.

Preferred sex steroid activity inhibitors result from substituting estrogenic nuclei such as those set forth in the foregoing examples with the preferred substituents set forth herein, including the side chains defined above (e.g.

-46- -Rl-[-B-R 2 Preferred sex steroid activity inhibitors in accordance with the invention include are not limited to: N-n-butyl-N-methyl-I 1-(16a-bromo-3',17'p-dihydroxy-es tra-1 0)-trien-7T cz-yl) undecanainide ("EM 105"): HOJO

(CH

2 0 00N(CH 3

)C

4

H

9 N-n-butyl-N-methyl-1 1-(1 6'a-brorno-3',lTa-dihydroxy-estra-11,3',51(10')-trien-7 a-yl) undecanam-ide ("EM 171"):

S(CH

2 10 C0N(CH 3

)C

4

H

9 47 N-n-butyl-N-methyl-l136'cx-chloro-3',17Tfrihydroxy-estra-l (1 0)-trien-7' ci-yl) undecanamide ("EM 139"): OH O

(OH

2 10 ON "H N-n-butyl-N-methyl-l1l6'ci-chloro-3',17a-dihydroxy-estra-1 ',3',5'(10')-trien-7 a-yl)undecanamide ("EM 170"): S.

S

S S cI 0 11

-OH

3

(OH

2 10 CN

,CA

-48yl) undecanamide ("EM 156"): HO"

(CH

2 1 0CON(CH 3

)C

4

H

9 N-n-butyl-N-methyl-1 1-(3'-hydroxy-17-oxo-estra-l',3',5'(1 O'),15'-tetraen-7'a-yl) umdecanamide ("EM 112"):

(CH

2 0 C0N(CH 3 )0 4

H

9 -49 N-n-butyl-N-methyl-1i-(3',I7frdihydroxy-17c-ethyny-estra-l ,3',5'(1O),15'-tetr aen-7cz-yl)undecanan-dde ("EM 123"): tC-=CH 4 1(CH 2 10 00N(CH 3 )0 4

H

9 aen-Tcx-yl)undecanamide ("EM 140"): HOJO

(CH

2 10 C0N(CH 3

)C

4

H

9 50 N-n-butyl-N-methyl-1 1-(3',17 dihydroxy-15',16'-methylefle-estra-lX3',5'(1 0), 1 5'-trien-7 a-yI)undecanamidde ("EM 136"):

(CH

2 10 C0N(CH- 3

)C

4

H

9 N-n-butyl-N-methyl-llI-(3',1dihydroxy-7-ethyny-estra-5',16'rmethylene-estra-l 0')-trien-7Ta-yl)undecanamide ("EM 138"): HOJO

(CH

2 0 C0N(CH 3

)C

4

H

9 -51 N-n-butyl-N-rnethyl-1 1-(3'-hydroxy-15',16'p-methylene-1 7'-oxo-estra- 1',3',5'(1O')-trien-7'a-yl)undecanamide ("EM 137):

S

*S

(CH

2 j 10 CON (CH 3

)C

4

H

9 N-n-butyl-N-methyl-ll1-(3'-hydroxy-16'-methylene-I7'-oxo-estra-l V3',5' trien-7ci-yl) undecanamide ("EM 175:

(CH

2

),QCON(CH

3

)C

4 Hg 52 N-n-butyl-N-methyl-llI-(3',17'p-dibenzoyl-14'p,15'0-epoxy-estra-1 0,36,59(10Q)t-rien-7'c-yl)undecanamide ("EM 180"): *0.00 .06.

0 0 0:0.0

C

6 H-SC0 2

(CH

2 10 00N(CH 3

)C

4

H

9 N-n-butyl-N-methyl-1 I-(3',17'p-dibenzoyl-14,5a-epoxy-estra-lV,3',5(1 trien-7'a-yl)unclecanamide ("EM 181"):

C

6

H-

5 CO2-

(CH

2 0 C0N(CH 3

)C

4

H

9 -53- N-n-butyl-N-methyl-1 l-(3,17'j-dihydroxy-,estra-1 0')-trien-7cxyl)undecanaxnide (TM 1081):

OH-

see*.

S.9.

see* .005 0:6S0S

-(CH

2 10 C0N(CH 3 )0 4

H

9 tridecynamide ("EM 163"): -54- N-n-butyl-N-methyl-14-(3',l~f dihydroxy-estra-',3',5'(10)-trien-7'a-yl)13tetradecynamide ("EM 195"):

(CH

2 1

,CON(CH-

3

)C

4 Hg Cs 0 *00 f..

N-n-butyl-N-methyl-8(3;l17frdihydroxy-estra-I ',3',5'CIO')-trien-Tcz-yl)7octynamidde ("EM 157T):

OH

C=-C(CH

2 5 CON (CH 3

)C

4

H

9 55 0 N-n-butyl-N-methyl-1 I-(6'-hydroxy-2'-(4"-hydroxyphenyl)-3ethyl-indol-N'-yl) undecanan-ide (EM 215):

(CH

2

)I

0 C0N(0H 3 4

H-

9 N-n-butyl-N-methyl-1 1-(6'-hydroxy-2'-(4"-hydroxyphenyl)-3',4'dihydronaphtalene-3'-yl) undecanainide: Hlj::C '(PCH 2 1 0 fCfON(CH 3

)C

4

H

9 -56- N-n-butyl-N-methyl-1 ,2-d iethyl-I ,2-ethanydyl) bis-phenol-3-yl)] undecanamide (EM 406): EM 406 6-hydroxy-2-(4'-hydroxyphenyl)-l-methyl-3-(6"-piperidino)hexyl-3,4dihydronaphatalene (EM 473) EM 473 lo1 N-n-butyl-N-rnethyl-llI-(6'-hydroxy-2'-(4"-hydroxyphenyl)-I '-methyl-3',4'dihydronaphatalene-3'-Yl) undecanamide (EM 690)

HO(CH

2 10 C0N(C 3

)C

4

H-

9 EM 690 -57- 6-hydroxy-2-(4'-hydroxyphenyl)-1-methyl-3-pentafluoropentylsulphinylnonyl.

3,4-dihydronaphatalene (EM 732) EM 732

;HASOC

5

H

6

F

2-(4'-hydroxyphenyl)-1 -methyl-3-(4"-(2"'-piperidinoethoxy)phenyl-3,4dihydronaphataene (EM 765) EM 765 O(CH2J 2 NCF1 10 2 -(4'-hydroxyphenyl)-l-methyl-3-[4"-(2"'-piperidinioethoxy)benzyl-3,4dihydronaphatalene (EM 431) EM 431 -58- N-pyrrolidinyl-1I -(7-hydroxy-3.-(4"-hydroxyphenyl)-4'-me thyl-2Hbenzothiopyran-2'-yl) undecanam-ide (EM 941) EM 941 HOS (CH 2 10 C0NC 4

HS

7 -hydroxy-3-(4'-hydroxyphenyl)-4-methyl-2-13'-(2'-piperidino)ethoxylpropynyl- 2H-benizopyraxi (EM 555) EM 555 0O(CH 2 2 NC1-1 1 N-n-butyl-N-methyl-llI-(7'-hydroxy-3'-(4"-hydroxyphenyl)-4'-methyl-2Hbenzopyran-2-yl) undecanamidde (EM 467) 59 7-hydroxy-3-(4'-hydroxyphenyl)-4-methyl-2-(6"-piperidino)hexyl-2Hbenzopyran (EM 721) YH EM 721 When a sex steroid nudeus is substituted with the side chain Rl[-B-R 2 L-G, it is preferred that the side chain have between about 7 and 30 carbon atoms and that L be separated from the nudleus by at least 3 intervening and preferably 6 atoms. In some embodiments, a polar moiety L or both) is preferably separated from the nucleus by at least 8 intervening atoms.

Additional inhibitors of the formula: Include but are not limited to those set forth below:

S

I T Inhfnitor EM-732 EM-473 EM-734 IEM-349

RI

-CH2- Absent -0- -0- Absent Absent 0 0 R2 (0-12)3 (C2)2 -(CH2)- 2 2 L G SO sH6Fs -Noo -N 0

-Q

-No Double bond Z Yes Yes Yes Yes yes 4CH2)- 4CH2)- 0 R6 -CH3 -CH3 CH3

H

R

3

-OH

-OH

H

-OH

RIO

-OH

-OH

OH

-OH

EM-428 0 -(CH2)2- 1 EM-384 0 -(CH2)3- 1 0 iCH3 H OH Yes 1 0 -CH3 H H EM-350 EM-357 EM-345 EM-371 EM-511 -0- 0 iCH2 T I 0 CH2 -C-Nr 0

-CNOH

2 C4H9q I 1 Yes Yes Yes Yes Yes o H I-OH 101 H I -OH 1-OH -CH2- -CH2- -CH2- Absent Absent Absent (CH2)2 (CH2)5 (CH2)4 3 2 2 ONC3 NCH3 so C4H9 C4H9 -sH6FS 0 0 0

H

H

H

-OH

-OH

:0H

-OH

-OH

-OH

-OH

61 2 I 0 -{CH2C31

O

I

0N CH3 I -OH

*OH

a.

a a a. a.

a a a a a a EM-547 1 I 0 -CH2)2 I NC2H5- C2H5 Yes S H H OH EM-762 -CH2- 4H) 2 4OC C4 H9 Yes S H OH OH EM-821 0 -{CH2)2 1 Yes NCH3 H OH OH EM-736 -CH2 Absent -(CH2)3- 2 wclb 3 C4H9 Yes -CH3 -OH -OH EM-698 Absent Absent -(CH2)4- 2 mNI3 C4H9 Yes -CH3 -OH -OH EM-721 -(CH2)6 Asr Absent 0 -NO Yes -CH- 3 -OH -OH EM-343 -01- 0- -(CH2)2- I -CD YES -CH3 *OH -OH -62- The following compound of the invention:

OH

CHO

EM 343 HO* O O(CHA)2NCsHio was synthesized and tested for its ability to inhibit the growth of ZR-75-1 human breast cancer cell line. The results are reported in Figure 5 as further discussed below.

0.0* The synthesis is described in Scheme A (page 66 herein) and Scheme 33 (page 183 herein). The synthesis of the compound 23 is described in Scheme A, and the starting materials and reagents were purchased from Aldrich Chemical Company Inc. (Milwaukee, Wis). Thus, the acd chloride 1 2 0 .0g; O.lmol) was added dropwise to methanol (60ml) at room temperature and with stirring. The solution was refluxed for Ih. The solvent was removed under reduced pressure and the resulting oil was dissolved in ethyl acetate. The organic solution was washed with saturated sodium bicarbonate aqueous solution. The organic solution was dried (MgSO 4 and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 1:9) to yield the compound 2 (Scheme A) (18.0g; 92%).

-63- The above ester 2 (21.2g; 0.108mol) and the nitrile 3 (available. from Aldrich Chemical Company Inc., Milwaukee, Wis.) (17.5ml; 0.129mol) were dissolved in benzene (750ml). The solution was refluxed and some benzene (100ml) was removed by means of a Dean Stark apparatus. The solution was left to cool down; then sodium ethoxide (9.2g; 0.135mol) was added. The resulting mixture was refluxed for 18h; it was washed with 1N hydrochloric acid aqueous solution. The organic solution was dried (MgSO 4 and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 1:4) to yield compound 4 (23g; (5 NMR; 300MHz; solvent CDC 3 standard: TMS) 3.78 (3H; s; OMe) 3.84 (3H; s; OMe) 3.95 (3H; s; OMe) 5.84 (1H; s; O-C-CH-CN) 6.43 (1H; d; J 25Hz; CH phenyl) 6.54 (1H; dd; J 2.5Hz and 8.5Hz; CH phenyl) 6.89 and 7.35 (2H; AB system; J 8.5Hz; CH phenyl) 7.79 (1H; d; J 85Hz; CH phenyl).

A solution of the ketone 4 (37.8g; 0.12mol) in acetic acid (400ml) and *concentrated hydrochloric acid aqueous solution (200ml) was kept at 90 0 C for 3h. The reaction mixture was neutralized with concentrated sodium hydroxide aqueous solution and it was extracted with ethyl acetate. The organic extract was dried (MgSO 4 and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 1:9) to yield compounds 5 17 .0g; 49%) and 6 6 .6g; -64- Triphenol 23 To a mixture of the ketones 5 (17.0g; 59.4mmol) and 6 (6.6g; 24.3mmol) was added pyridine hydrochloride (90g). The mixture was heated at 220 0 C for 1N hydrochloric acid aqueous solution (250ml) was added and the resulting mixture was extracted several times with ethyl acetate. The organic extract was dried (MgSO4) and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 3:7) to yield compound 23 (14.1g; 69%).

The continuation of the synthesis of EM 343 is described below with reference to Scheme 33 (page 183).

Diether 24 To a mixture of the triphenol 23 (14.1g; 57.8mmol) in 3,4-dihydro-2H-pyran (200ml) at 0°C and with vigourous stirring was carefully added p- .toluenesulfonic acid monohydrate The reaction mixture was stirred at 0°C for a further Ih. Ether (300ml) was added and the solution was washed with IN sodium hydroxide aqueous solution. The organic extract was dried (MgSO 4 and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 1:9) to yield compound 24 23 .4g; (5 NMR; 300MHz; solvent: CDCl 3 standard: TMS) 1.5-2.1 (12H; m; O-CHH-CH 2

-CH

2

-CH

2 -0 THP) 3.55-3.65 (2H; m; O-CH-CH 2

CH

2 -CH2-CH 2 -0 THP) 3.75-3.95 (2H; m; O-CH-CH 2 -CH2-CH 2

-CH

2 -O THP) 4.16 (2H; s; Ph-CH 2 -CO) 5.40 (1H; t, J 3Hz; OCH-CH 2

-CH

2

-CH

2

-CH

2 -0O THP) 5.49 (1H; t; J 3Hz; O-CH-CH 2

-CH

2

-C

2

-C

2 -0 THP) 6.55 (1H; dd; J 2.5Hz and 8.5Hz; CH phenyl) 6.61 (1H; d; J 2.5Hz; CH phenyl) 7.03 and 7.17 (2H; AB system; J CH phenyl) 7.77 d; J 8.5Hz; CH phenyl) 12.60 (IH; s; Ph-OH).

Chroman-4-one 25 and Chalcone 26 (R=H) To a mixture of the diether 24 2 4 .4g; 59.2mmol) and the aldehyde

(OHCC

6

H

4 0H) 7 .6g; 62.18mmol) in dry benzene (750ml) was added piperidine (500pl). The solution was refluxed for 48h and water was continuously removed by means of a Dean Stark apparatus. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 1-9) to yield compounds 25 (143g; 47%) and 26 (8.4g; This last compound can be converted to compound 25 (R=H) *by heating with sodium acetate in methanoL Chroman-4-one 25 (R=H) (8 NMR; 300MHz; solvent CDCI 3 standard: TMS) 1.5-2.1 (12H; m; O-CH-CH 2

-CH

2

-CH

2

-CH

2 -O THP) 3.45-3.65 (2H; m; O-CH-CH 2

CH

2

-CH

2

-CH

2 -O THP) 3.8-3.95 (2H; m; O-CH-CH 2

-CH

2

C

2

-CH

2 -0O THP) 4.05-4.1 (1H; m O-CH-CH-C=--O) 5.25535 (IH; m O-CH-CH-C=--O) 5.35-5.55 (2H- m; O-CH-

CH

2

-CH

2

-CH

2

CH

2 -0 THP) 6.6-7.1 (10OH; m; CH phenyl) 7.85-7.95 (1H; m; CH phenyl).

-66-

SGHEMEA

MeG 0 cI Meo" Meo 0 N. OMe MeoG a a a MeG 0 ome RO 0 R=Me MeO 6, R--H MeG'

HO'

-67- EM 343 A mixture of compound 25 (1.90g; 3.8mmol), 1-(2-chloroethyl)piperidine hydrochloride CI(CH 2 2 NCsH1o. HCI (1.18 g; 6.5 mmol) and sodium carbonate (0.97g; 9.1 mmol) in acetone (100 ml) was kept under reflux and with stirring for 48h. The precipitate was filtered off and washed thoroughly with acetone. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel (hexanes:acetone; 7:3 a few drops of triethylamine) to yield compound 25 (R=(CH 2 2 NC5H 1 0) (1.

5 7 g; To a solution of compound 25 (R=(CH 2 2 NCsH 1 o) (90mg; 1431mol) in ether was added methylmagnesium iodide (3.0M solution in ether; 1.2ml; 3.6mmol) at 0"C and with stirring. The mixture was stirred for a further 3h. at room temperature, then washed with saturated ammonium chloride solution. The organic solution was dried (MgSO4) and the solvent was removed under reduced pressure. The residue was quickly filtered through silica gel (ethyl acetate: acetone; 1:1) to afford compound 27 (R=(CH2) 2 NCsHio, Rc CH3)(90mg; 97%) which was used directly in the next reaction.

A solution of compound 27 (R=CH2) 2 NCSHo1, Rc CH 3 (90mg; 139tmol) in a mixture of acetic acid (60ml) and water (6ml) was kept at 100 0 C for 10min. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel (ethyl acetate: acetone; 3:2) to yield compound 28 (EM 343, R=(CH 2 2 NCsHlo, Rc CH 3 (40mg; (5 NMR; 300MHz; solvent: CD3OD; standard: TMS), 1.46 (2H; m; cyclo-N-CH2-CH2-CH 2

-CH

2

-CH

2 1.60 (4H; -68m; cyclo-N-CH 2

-CH

2

-CH

2

-CH-

2

-CH

2 2-02 (3H; s; CH 3 2.56 O4H; m; cyclo-N- CH2..CH 2

-CH

2 -CH2-CHZ-) 2.76 (2H; t, J 5Hz, O-CH 2

-CH

2 4.06 (2H; t; J 5Hz; C)-

CH

2

-CH

2 5.77 (1H; s; O-CH-Ph) 6.12 O1H; d; J 2Z5Hz; CH Phenyl) 6.35 O1H; dd; J 2.5Hz,8Hz; CH Phenyl) 6.70 (2H; d; J 8_5Hz; CH Phenyl) 6.77 (2H; d; J 8.5Hz; CH Phenyl) 6.98 d; J 8.5Hz; CH Phenyl) 7.12 (11-L d; J 8Hz; CH Phenyl) 7.19 (2H; d; J 8.5Hz; CH Phenyl). Mass Spectoscpy-. M+ 459.

The product, EM 343, was then prepared for efficacy testing using the ZR-75-1 human breast cancer cell line.

Maintenance of Stock Cell Cultures ZR-75-1 cells (83rd passage) were obtained from the American Type Culture Collection (Rockville, MID) and routinely cultured in phenol-red free RPMI 1640 supplemented with I nM E 2 2 mM L-glutaxnine, 1 mM sodium pyruvate, m.M N-2-hydroxyethyl-piperazine-N-2-ethanesulfonic acid, 100 LU **penicillin/mi, 100 jig streptomycin/mi, and 10% fetal bovine serum (Hyclone, Logan, UT) under a humidified atmosphere of 95% air, 5% C0) 2 at 37 All media and medium supplements were purchased from Sigma. Cells were subcultured weekly by treatment with a pancreatic solution containing 0.02% EDTA The cell cultures used for the experiments herein described were between passages 89 and 94.

69- Measurements of Cell Proliferation Cells in their logarithmic growth phase were harvested, briefly centrifuged, and resuspended in RPMI 1640. Cells were then plated in triplicate in LIMBRO 24-well plastic culture plates (2 cm 2 !/well). Since plating density influences the effect of hormones on ZR-75-1 cell growth, cells were plated at a density of 1 x 104 i0~cells/well. After 72 h, medium was replaced with fresh medium of identical composition containing, in addition, the concentrations of steroids and/or inhibitors EM 312 or EM 343) indicated along the X-axis of Figure Control cultures received the ethanol vehicle only. Cells were then allowedl to grow at 37 0 C for 10 days with medium changes (of identical composition) every 2 days. In absence of inhibitors, in 0.21 nM estradiol (E2)-containing medium, :ZR-75-1 cells have doubling time of about 48 It After E 2 and/or antiestrogen treatment, cells were harvested by addition of ml of a pancreatin solution (Sigma) for 5-10 min at 37 "C before addition of ml of RPM] 1640 containing 5% dextran-coated charcoal-fetal bovine serum in order to block enzymatic action. Cell number (0.10-mld aliquot) was determined by measurement of DNA content as previously described (Simard et al., Endocrinology 126: 3223-3231, 1990).

As may be seen from Figure 5, EM 343 provided extensive cell growth inhibition at low concentration. Half-maximal effectiveness occurred at a concentration of 2.55 x 10 10 M. Without intending to be bound by theory, it is believed that the B-ring alkyl substitution utilized in certain preferred embodiments of the invention the methyl substitution of EM 343) enhances effectiveness relative to compounds lacking such a substitution. For example, another species of the invention.

SOH

HO 0. 0EM 312 O(CH2)2N reported by Saeed et al Med. Chem. 33: 3210-3216, 1990) and Sharma et al. (J.

Med. Chem. 33: 3216-3222 and 3222-3229, 1990) to have antiestrogenic activity S. underperformed EM 343 in comparative testing in our laboratory on the growth of human ZR-75-1 breast cancer cells, the ICso value of EM-312 being 3-fold higher at 8.43 x 10 1 0 M (Fig. Set forth below are some flow charts description and illustration of a number of preferred synthesis schemes for certain preferred antiestrogens in accordance with the invention. The steps set forth below are set forth merely by way of example. Those of skill in the art will readily recognize alternative synthetic 71 pathways and variations capable of producing a variety of antiestrogens and other sex steroid activity inhibitors in accordance with the invention.

0 0 0*00 0 0 0 000 0 00*0 00 0 a.

0*00*0 a -72- EXAMPLES OF SYNTHESIS OF PREFERRED INHIBirORS OF SEX STEROID ACTIVITY Instrumentation The IR spectra were taken on a Perkin-Elmer 1310 spectrophotometer. Proton NMR spectra were recorded on a Varian EM-160A (60 MHz, when specified) or a Varian XL-200 (MHz) instrument The following abbreviations have been used:s, singlet, d, doublet; dd, doublet of doublet; t, triplet, q, quadruplet;, and m, multiplet. Chemical shifts are reported in i values in ppm relative to tetramethysilane (TMS) as internal standard. Mass spectra (MS) were obtained on a V.G. Micromass 16F machine. Thin-layer chromatography (TLC) was performed on 0.25 mm Kieselgel 60F254 plates Merck, Darmstadt, FRG). For flash chromatography, Merck-Kleselgel 60 (230-400 mesh AS.T.M.) was used.

•000 All solvents used in chromatography have been distilled. Unless otherwise noted, starting material and reactant were obtained commercially and were used as such or purified by standard means. All solvents and reactants purified and dried were stored under argon. Anhydrous reactions were performed under an inert atmosphere, the set-up assembled and cooled under argon.

Organic solutions were dried over magnesium sulfate, evaporated on a rotatory evaporator and under reduced pressure. Anhydrous solvents were prepared in the following way.

-73- SOLVENT DISTILLED OVER AMINE, DIMfETHYLFORMAMIDE CaH2 HEXANE, DICI{LOROMETH-ANE P 2 03 ACETONE K2C03 BENqZENE LIAIH4 TOLUENE Na ETHER,, TETRAHYDROFURAN LiAlH4, Na benzophenone

OS

gee.

0 900 C 0e0* 0**0

C

OSOO

C. Ce C 0

C

0000

C

.0Cc 4 0 0000 0 0*0 0 C, 0 0 *000 *0 0 00 C 90

C

000000 0 LIST OF ABBREVIATIONS Bz Benzyl DMF Dimethylformamide EDTA Ethylenediaminetretraac-tic acd HMF1A Hexamethylphosphoramidde HPLJC High pressure liquid chromatography IDA Lithium diisapropylamine mCPBA meta-chloroperbenzoic acd mom Methyloxymethyl NAD Nicotinamide Adenine Dixiudeotide NADH- Nicotinamidde Adenine Dinucleotide reduced form PTSA Para-toluene sulfonic add PT'SC1 Para-toluene sulfonyl chloride TBDMS t-butyldimethylsilyl TI-F Tetrahydrofuran THIP Tetrahydropyrannyl TMS Tetramethylsilyl 74 EXAMPVLE I Instrumentation IR spectra were obtained in a Perkin-Elmner spectrometer 1310. LTV spectra were recorded in methanol on a Beckmnan DU-6 spectrometer. H-NMR spectra were obtained at 200 M[Hz on a Varian XL-200 spectrometer. Chemical shifts are reported in ppm units with tetramethylsilane as internal standard. Mass spectra were obtained on Micromass 16F spectrometer.

N-butyl, N-methyl- 2,1 3-Bis (4-hydroxyphenyl)-12-pentadecenoic amide (EM-142, compound 5 withi The synthesis of thids compound is described in the scheme I (infra) where 12.1 3-Bis (4-methoxyphenyl)-1 1-petadecenoI (3) 4'-methoxy-2-ethyl,2-(4-methoxyphenyl). acetophenone (710 mg, 2-5 mmol, prepared from desoxyanisain, ethyl bromide and LIDA by a known method) in THE (10 ml) were added, under argon, to Grignard reagent prepared from 1l-bromo-tetrahydropyranyl undecanol (6.6 g, 19.7 inmoles) and magnesium (0.6 g, 24.7 mmoles) and THF (10 ml). The mixture was stirred for 18 hours, then acidified with IN HC1 and extracted three times with ether. The organic phase was washed with water dried over anhydrous magnesium sulfate, and evaporated under reduced pressure. The residue was chromatographed on Silica-gel (Kieselgel, 60F254, Merck, 0.063-0.200 mm, 100 Elution with a mixture of hexane-ethyl acetate (9:1 v/v) gave 12,13-Bis-(4-methoxyphenyl)tetrahydropyranyl pentadecan-1,12-diol (991 mg, 76%) as a mixture of diastereoisomers; colorless oil, IRvmax (neat) 3480, 1600 cm- 1 'H-NMR (6, CDCl 3 0.62 (3H, t, J= 73 Hz, CH.CH 3 2.73 (IH, 2d, J=9.7 Hz, -CHCH 2

CH

3 3.25-4.00 (4H, m, -CH2OCHOCH 2 3.76 and3.79 (6H, 2s, -OCH 3 4.57 (1H, t, J=1.1 Hz, -02-CH-CH 2 and 6.71-7.30 (8H, mH-Ar)ppm. MS m/e= 523 (M+-H 2 0).

The above diastereoisomers (920 mg, 1.8 nmmunol) dissolved in methanol (30 ml) and 5N HCI (5 ml) was refluxed for 1 hour, then collected, and extracted three times with ether. The organic layer was washed with water, dried over anhydrous magnesium sulfate, evaporated under reduced pressure to dryness and finally chromatographed on silica gel (Kiesegel, 60F254, 0.063-0.200 mm, Merck, 100 Elution was a mixture of hexane-ethyl acetate (7:3 v/v) gave 1 2 ,13-Bis(4-methoxyphenyl)-11-pentadecenol (710 mg, 65% from compound colorless oil, IR, vmax (neat), 3340, 1600, 1030 cm- 1 UV Tmax (log E) 231 (4.27) nim; 'H-NMR CDC1 3 0.88 (3H, t, J 7.3 Hz, -CH 2 CH3), 3.30 (1H, t, J= 9.7 Hz, -CH-CH2,CH 3 3.63 (2H, t, J 6.6 Hz, -CH 2 0H), 3.76 and 3.78 (6H, 2s, -OCH 3 -76 5-51 (IH, t, J= 8.8 Hz, 1 -C CHi-) and 6.63-7.10 8H, n, H-Ar)*ppm; MS m/e 438 N-bu tyl. N-methyl .12,13-Bis (4-methoxyphenyl)-1 1 -Tntadecenoic amnide (4) To a cooled solution of alcohol 3 (710 mg, 1-56 inmol) in acetone (17 mld) was added Jones' reagent (8N-chromnic acid solution, 0.77 ml). After 30 minutes, isopropanol (5 ml) was added and the midxture was poured in water and extracted three times with ethyl acetate. The organic layer was washed twice with brine, dried over magnesium sulfate and evaporated to dryness. The crude 12,13-Bis (4-methoxyphenyl)-11-pantadecenoic acdd was used in the next step without purification. To its solution in anhydrous methylene chloride (4 :nml) at -10 0 C was added, under stirring, tisobutylarnine (470 iii, mmol) and isobutylchloroformate (280 ILI, 2.1 mmol). After 40 minutes, N-methylbutylamidne (1-5 ml) was added and the mixture was stir-red at room temperature during 1 hour. Methylene chloride (50 mld) was added. The organic solution was washed with IN HCI, saturated sodium bicarbonate solution and water dried on magnesium sulfate and evaporated to dryness. The residue was purified by "Flash chromatography" on silica gel (Kieselgel 60, Merck, under 0.063mm, 50 Elution with a mixture. of hexane-ethyl acetate (4:1 v/v) gave N-butyl, N-methyl-12,13-Bis -77- (4-methoxyphenyl)-1-pentadecenoic amide (549 mg, 68%) colorless oil; IR vmax (neat), 1640, 1600 cm-1; UV Tmax 00(log e) 230, (439) nm; 1 H-NMR CDCl 3 0.85-0.98 (6H, m, 2-CH 2 CI3), 2-27 (2H, t, J= 7.1 Hz, CH 2 CON), 2.91 and 2.%96 (3H, 2s, -NCHl3), 3.25-336 (3H, m, -NCH and CH 3 CH2CI-), 3.77 and 3.78 (61-H, 2s, OCHj 3 5.50 (IH, J= 7.1 Hz, and 6.69-7.01 (8H, m, H-Ar) ppm; MS m/e 521 N-butvl. N-methyl-12.13-Bis (4-hydroxyphenl)-12-pentadecenoic amide (EM-142, compound 5 with x To the above dimethoxy amide 4 (117 mg, 0.22 mmol) in CH 2 Cl 2 (1 ml) at OC was added, under argon, 1.0 M borane tribromide (675 tl). The solution was *stirred for 1 hour, then poured into water and extracted with ether The organic solution was washed with water, dried on magnesium sulfate, and evaporated to dryness. The residue was purified by "Flash chromatography" on silica gel (Kieselgel 60, Merck, under 0.063 mm, 30 Elution with mixture of hexane-ethyl acetate (4:1 v/v) gave N-butyl, N-methyl-12,13-Bis (4-hydroxyphenyl)-12-pentadecenoic amide (EM-142, compound 5 with x (34 mg, colorless oil, IR vmax (neat) 3300, 1600 cm-1; UV Ymax (log E) 235 (4.25) rn; H-NMR CDC13), 0.76 (3H, t, J= 7.3 Hz, -CH 2 CH3), 0.96 (3H, t, J= 7.3 Hz, N (CH 2 3 C13), 2.05-2.20 (4H, m, CH2-C C-CH 2 235 (2H, t, J= 7.0 Hz, 78

-CH

2 CON-), 2.97 and 3.00 (3H, s, -NCfi3), 3.29 and 3.41 (2H, 2t, J= 7.3 Hz, and 6.59-7.09 (8H, m, H-Ar) ppm; MS m/e 493 -79- Scheme 1 o 1) LDA 2) CA~Bra xz 2 1) B('M9(CHJ 1 1

IOTHP

2) H 3) Ct- 3

NHC

4

H

9

CH-(CH

2 )xOH CH(CH 2 1

CON(CH

3

)C

4

H-

9 4 (CH2)xCON(CH3)C4H9 EXAMPLE 2 EFFECTIVENESS OF ANTIESTROGEN SYNTHESIZED IN EXAMPLE 1 EM-142 from example 1 was tested by competition binding on the rat uterine cytosol estrogen receptor with PH] estradiol according to Asselin et al. (1976) prodecure. Fig. 1 shows the binding affinity of EM-142 compared with estradiol, diethylstilbestrol and ICI 164384 (Wakeling and Bowler, 1987). (incubation was performed at 25 0 C for 3 hours). It can be seen that EM-142 is only 3 times less potent than 17p-estradiol itself while being more potent than the antiestrogen ICI 164384.

The antiestrogenic activity of EM-142 was measured in vivo by inhibition of the o estradiol-induced stimulation of uterine weight in adult female ovariectomized Balb/c mice (body weight 19-20 g) sacrificed five days after ovariectomy. EM-142, and/or estradiol contained in ethanol were injected subcutaneously in the appropriate groups in a solution of 0.9% sodium chloride and 1% gelatin at different concentrations in 0.2 ml for EM-142, twice daily, starting on the day of ovariectomy for a total of 9 injections.

Estradiol was injected at the dose of 0.01 ,g in 0.2 ml, twice daily, starting on the morning after ovariectomy for a total of 8 injections.

After sacrifice, the uteri were rapidly removed, freed from fat and connective tissue and weighted. Results shown in Fig. 2 are the means SEM of groups of -81- 9-10 mice. It can be seen that the very low dose of 03 tg already has a significant inhibitory effect of an E 2 -induced uterine growth and that a complete reversal of E 2 effect is obtained at higher doses. A half-maximal inhibitory effect is in fact observed at approximately 3 tig while, at the doses used, 20 tpg causes a complete reversal of estrogenic action. Note that EM-142 has no estrogenic effect on uterine weight, thus demonstrating its potent antiestrogenic activity.

e a -82- EXAMPL-E 3 Scheme 2 N-butyl. N-methyl-12,13-Bis-(4-hydroxyphenvl) Rentadecanoic amide (6) ~ID H 2 CafafYtic HO (CH2)xCON(C-i 3

)C

4 -I (1)xCON(CH)CIg 6 -83- ExAmPLE 4 Scheme 3 N-butyl, N-methyl-5--f4-(2-[4-hydroxyphenylI berzothiophen-3-l) fonnvll p2henoxyl hexanoic amide (12) 4.

S S 4 4@SO

OSSS

St 45 4 0 4 4 4*5

S

Sr..

4* C 5 7 1) BuU 2) p-fC 6

H

4 OC4- 3 cS)-a/OCH 3 8

AIC

(CH

3 3 siI 0 0S00.S S S 1Br(C4 2

)S

Na0' i 2 5

CON(CH

3

)C

4 Hq 0

O(CH

2 5 C0 2

H

1CICO04-6Bj 7 12) BafZC.CH 9 S OCH 3 Cl H C

I

-8&4- EXAMPLE 5 (Scheme 4) N-butvl. N-methyl-64fp-(trans-l' 2'-bis W4-hydroxyphenl)-1 '-butenyl phenoxvyll heptanoic amide (18) 1) BrMgC 6

H

4

OCH

2

C

6

H

5 i 2) H{+ 13

-OCH

3 SNz. NaH

CH

3 0 N. BrCHA)

OH

0CH 3 16 2) CICOi-Bu 3) NH(CH 3

)C

4

H

9 2)6--(3 H 0 0-1

-OCH

3 oo J I BBr 3

:S(CH

3 2

CH

3 0

CICH

2

CH

2

CI

O(CH

2 6

CON(CH

3

)C

4

H

9

O(CH

2 )6CON(CH 3

)C

4 Ne 85 EXAMPLE 6 Scheme N-buty.Nmtvl'bdoy 2 4 -hdroxyhenyl)3..meth dindol-N.'.y 1 undecanoic amide (22) a. a a.

a a a. a a a a.

a

CH

3 0

CH

3

CH

3 0 N N /OCH 3 H 1) NaHIDMF 19 2) Br(CH 2 1 1. 1 H O- Bu 3) NH(CH 3

)C

4

H

9

CH

3 O BBr 3 -86- ExAJm.PLE7 Scheme 6 N-butyl N-Methyl-i1-F4,4'-(1 .2-diethyl-1 .2-ethanydiyl) bis-p2henol-3-ylI undecanoic amnide (28) a a a. TBDMSCI (I eq)

DMAP

TBDMSO'

/KCONEI

2

K

2 C0 3

CN

3 COCH4 3

A

a.

a a I 1) BuU I eq), THF TMEDA-78*C 2) 8rqCHOI 0 F 1)

H.

2) NaOH- 10 C0N(CH 3

)C

4 Hg 27

NH(CH

3 )nei 2) NaOH 87 EXAMPLE 8 (Scheme 7) N-butyl, N-methvl-[6'-hydroxy-2 I (4-hydroxyphenyl)-(lv. 2'-dihvdronaphta-len- T-yl) undecarioic amide (34)

CH

3 oaO' 6-methoxy-2-tetraion R CO 2 Et

CH

3 0O NaHI(Eto) 2 00 Na H RBr (1 eq)

CH

3 0 O NaH Br(CH 2 1 0 R CO 2 Et

CH

3 O4 (H)0.O 31 0) Na 2

CO

3

A

B

2

(CH

2 10

-Q-

R= CH 3

C

2

H

5

C

3

H

7 1) BrMgC 6

H

4

OCH

3 2) H' 3) OH-

CH

3 0. '(CH 2 10 C00H 1) BBr 3 2) CICO i-BuN(HC, R -OH 3) NaO NHC) 4

H

:1 Nz HO NI (CH 2 10 C0N(CH 3

)C

4

H

9 -88- EXAMPLE 9 SYNTHESIS OF A STARTING COMPOUND, N-N-BUTYL, N-METIHYL-11-(3'- BENZOYLOXY-17-OXO-ESTRA-1',3',5(10')-TRIEN- a-YL) UNDECANAMIDE (9) (SCHEME 8, INFRA)) 19-nor-testosterone-acetate 3-enolacetate (2) In an apparatus supplied with a drierite drying tube, a solution of 19-nor-testosterone (10) (100 g; 0365 mole) in acetic anhydride (200 ml), pyridine (32 ml) and acetylchloride (320 ml) was heated at reflux under magnetic stirring, for 3 h and then concentrated to dryness under vacuum. The dry residue was triturated in absolute ethanol, filtered and washed with little portions of absolute ethanol. After drying, 19-nor-testosterone acetate 3-enolacetate was obtained as a white powder (121.4 g, yield 93%) mp. 176-177*C. The structure was confirmed by spectroscopic means.

1 78-acetoxv-estra-4,6dien-3-one (3) To a cooled suspension of enolacetate (121 g; 0337 mole) in a mixture of DMF (330 ml) and water. (7.2 ml) at 0 0 C was added, under nitrogen, over a period of 1 h, N-bromosuccinimide (63 The resulting solution was stirred for an -89 additional 0.5 h at 0 0 C. Then lithium carbonate (60.8 g) and lithium bromide (30.4 g) were added. The mixture was heated at 95*C for 3 h and then poured into 1.7 1 of ice-cold water containing 165 ml of glacial acetic acid. After stirring during 15 hours, the crude 17p-acetoxy-estra-4,6-dien-3-one was filtered, washed with water, dried in a desiccating apparatus and recrystallized twice from isopropyl ether (72 gyield 68%, mp 110'C). The structure was confirmed by spectroscopic means.

*I .7 aeo5 sra4e n 7c11om-cton-ecl 10 ga38 toy wstas-enssolved 4) y te (6 n A.e Psoplrtion waf cooledto and sintsnieH0bah oti slto a To this* bomtudeaoltiao pyrf y et-d hr-Hp n(99&4- l rsl 9.dstle in drbryonethor (10 m,38ml) was dssolvaerid if dry ether (768 solutian was then stirred over a period of 16 hours at room temperature. Afterwards, sodium bicarbonate was added to the mixture. The residue was filtered and the solvent was evaporated under vacuum.

The product was then filtered through basic alumina (250 g, Woelm, grade II) using petroleum ether (30-60) as solvent (112 g, 81%).

B. Grinard reagent *o In a dry three-neck flask (1000 ml) under dry argon, magnesium (12.0 g, 494 mmol) was placed and activated with iodine. Magnesium was heated with the flame to remove iodine and to dry the apparatus. The system was then cooled to -20 0 C, and a solution of 11-bromo-undecanol tetrahydropyranyl ether (73.8 g, 211 mmol) in dry THF (420 ml) was added dropwise. The mixture was stirred under dry argon during one day at The mixture was cooled to -35°C (±2 0 C) using a dry ice/CC4/acetone bath. The anhydrous cuprous chloride (1.18 g, 12 mmol) was added and the mixture was stirred over a period of -91- C. Addition of Griynard reagent After 0-5 h, using the same apparatus mentioned above (Ar, a solution of 17p-acetoxy estra-4,6-diene-3-one (32.0 g, 102 nunmmol) in dry THF (300 ml) was added dropwise over a period of 6 h to the Grignard reagent (red coloration appeared and disappeared). The mixture was stirred for an additional 1 h and, after removal the cooling bath, acidified (about 0°C) with acetic acid (40 ml), ooo diluted with water and extracted with ether The ether solution was washed with a saturated sodium bicarbonate solution and water. The organic layer was dried over anhydrous magnesium sulfate and evaporated under reduced pressure to dryness.

The residue was dissolved in MeOH (660 ml) and 5N HC1 (180 ml), refluxed for 1 h and 45 mrin, then concentrated under reduced pressure and cooled in an ice bath. The mixture was then filtered to remove the white precipitate. After the i solution had been diluted with water and extracted with methylene chloride the organic layer was dried over anhydrous MgSO4 and evaporated under reduced pressure to dryness. Finally, the product (55.9 g, brown oil) was chromatographed on silica gel (Kieselgel 60F254, Merck, 0.063-0.200 mm, 1500 g).

Elution with mixtures of methylene chloride and ethyl acetate (4:1 to 1:2 v/v) and then pure ethyl acetate gave crude 7a-(11'-hydroxy-undecyl)-17p-hydroxy estra-4-en-3-one (34.8 g) which was dissolved in dry pyridine (200 ml) and dry -92acetic anhydride (200 ml), stirred 17h at room temperature and then poured in ice-water. The product was extracted with methylene chloride washed with IN hydrochloric acid, water, saturated sodium bicarbonate and water (3X), dried on anhydrous magnesium sulfate and filtered. After evaporation of solvent, the mixture (35 g) of 7a- and 7p-diacetoxyenones and degradation products of Grignard reagent were separated by flash chromatography on silica gel (Kieselgel 60, Merck, 230 mesh ASTM, 2.0 kg) developed with a mixture of hexane and diethyl ether (2:3 The first product eluted was pure amorphous 7 a-(11'-acetoxy undecyl) 17 pacetoxy-estra-4-en-3-one (20.8 g, 39.4 mmol, yield from dienone was Further elution gave the 7p-isomer (5.4 g, 103 mmol, All structures were determined by spectroscopic means.

Z-(11'-hydroxy-undecyl) estra-1 3,5(10)-trien-3,7l diol (6a) Under dry argon, a solution of 7a-(11'-acetoxy undecyl) 170-acetoxy-estra-4-en-3-one (17.0 g, 32.4 mmol) in dry acetonitrile (150 ml) was added rapidly to a suspension of cupric bromide (14.8 g, 66.2 mmol) and mmol) and lithium bromide (2.89 g, 33.6 mmol) in warm acetonitrile (75 ml).

The mixture was heated to reflux over a period of 30 min and stirred vigorously, and then cooled to room temperature. A saturated aqueous solution of sodium bicarbonate (50 ml) was added, and then the organic solution of sodium bicarbonate (50 ml) was added, and then the organic -93compound was extracted with ethyl acetate O3x 150 rnl). The organic layers were washed with water, dried over anhydrous magnesium sulfate, filtered and evaporated under vacuum to dryness. The residue was chromatographed on silica gel (Kieselgel 60F254 Merck 0.063-0.200 mam; 1000 Elution with hexane-ethyl acetate (1:1 v/v) gave the 7cx-(11'-acetoxy-undecyl) estra- V,3',5'(10')-trien-3,1y7..diol, 17p-acetate (6b) (8.51 g; 50.3%) and the starting product (1.33 g; The above diacetate phenol (8-51 g, 16.2 mmol) was dissolved in methanol ml) and sodium hydroxyde 30% (wfv) (9 mld). The mixture was refluxed for min under dry nitrogen. The solution was then concentrated under vacuumand diluted with hydrochloric acid The mixture was extracted using ethyl acetate (4 x 150 mld) and the ethyl acetate extract was washed with water, dried over anhydrous magnesium sulfate, filtered and evaporated under vacuum. The evaporation gave 7a-(11'-hydroxy undecyl) estra-1,3,5(10)-trien-3,17p-dioI (6a) (6.99 g, 98% brut) as a yellow foam, the structure of which was confirmed by spectroscopic means.

3-benzoyloy 7c-(11'-hydrox'v undecyl) estra-13,5(10)-trien-17ft-ol_(7 The above triol (6.99 g; 15.8 mol) was dissolved in acetone (25 ml) and an aqueous solution of sodium hydroxyde (IN, 19.1 ml). The mixture was cooled -94to 0 C using an ice/water bath. Benzoyl chloride (2.22 ml, 19.1 mmol) was then added dropwise. The mixture was stirred for 40 min at 0oC and then diluted with water. The solution was extracted using ethyl acetate (3X) and the organic layers were washed with a saturated aqueous solution of sodium bicarbonate and finally with water. The ethyl acetate solution was dried over anhydrous magnesium sulfate, filtered and evaporated under vacuum to dryness. Then, the residue was immediately chromatographed on silica gel (Kieselgel, 60F254, 0.063-0.200 mm; 500 The chromatography was carried out, first, using methylene chloride as solvent (about 1 liter) and secondly the pure 3-benzoyloxy 7a-(11'-hydroxy undecyl) estra-l,3,5(10)-trien-17-ol colorless oil (6.50 g, 75%) was eluted with methylene chloride-ethyl acetate (5:1 about 1 liter and 4:1; The structure was confirmed by spectroscopic means.

1-(3-benzoyloxy-17'-oxo-estra-1'3'5'(10)-trien-7'-yl) undecanoic acd (8) 9 To a cooled solution of 3-benzoyloxy-7a-(11'-hydroxy undecyl)estra-1,3,(10)trien-173-ol (43 g) in acetone (100 ml) was added dropwise Jone's reagent (8N-chromic acid solution, 6.7 ml). After 30 min, isopropanol (40 ml) was added and the mixture was concentrated under vacuo. Water was added and the mixture was extracted four times with ethyl acetate. The organic layers were washed twice with brine, dried over magnesium sulfate and evaporated to dryness. The crude I l-(3'-benzoyloxy-17'-oxo estra.~1',3',5'(1 O')-trien-7'ct-yl) undecanoic acid (3.94 g) was used in the next step without purification.

0S S S 0

.SSS

S

S. 55

OS*S

0e*e S. SO S S

S

055@

S

5*59 55

S

550 S *0*b

S

@9

S.

0* 0 *OSO.0

S

-96 Scheme 8 0 It

JC-CH

3 0 0 Is 0C- ci- 0 of IC0CR 3 4 7-pH 7-aH

CH-

2 0 H

CGHSCO

2 6a R-H 6b R=OCOCH 3 0

C

6 1-1 5 C0 2 10 C0 2 1 2 10 CONMeBu 9a C 6 HS00 9b Ra=H -97unidecanamide (2b) To 11 3 -benzoyloxy-17'-oxo-estra-i '):-trien-7'cx-yl) undecanoic acid (8) (3.94 g, 7.22 n-nol, dissolved in anhydrous CH 2 Cl 2 (100 ml) and cooled at -10 0 oC was added tributylamnine (2.18 ml, 9.15 mmol) and isobutyichioroformate (1.30 ml, 10.0 nunol). The solution was stirred during 35 min. and Nmethylbutylarnine (13 ml, 109.7 nunol) was added. The mixture was warmed *to room temnper-ature and stirred during 1 h. Afterward, CH 2

CI

2 was added and the organic phase was washed with 1N HCI, water, saturated sodium bicarbonate solution and finally with water, dried with anhydrous MgSO4 and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel. Elution with mixture of EtOAc/hexane (1-5:8-5 v yielded N-butyl, N-methyl-I1-(3'- benzoyloxy-17'-oxo-estra.. trien-7'a-yl) undecanamide (9a) (4.25 g, 96%) as colorless oil; IR v (neat) 1750, 1725 and 1640 cm- 1 The above described benzoyloxy amide (341 mg, *.0.54 mmol) was dissolved in methanol (10 ml) and cooled at 0 0 C. Following this 2N NaOH (5 mld) was added and the mixture was stirred during 60 mini. at 0 0 C The solution was neutralized with 1N HC1 and extracted with CH 2 Cl 2 The organic phase was dried with anhydrous MgSO 4 and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel. Elution with mixture of EtOAc/hexane (3:7 v/v) yielded N-butyl, N-methyl-i I-(3'-hydroxy-1 7'-oxo,estra-1 (1 0)-trien-7'ac-yl) undecanamide (9b) (284 mg, 97%) as colorless oil; I H-NMR 5 (CDCi1 3 0.91 (s,3H,l 8'-CH 3 2.76 -98 app(d,1HJ=16,~-iz part of ABX system, 296 and 2.98 (2s,3HN-CH 3 3.27 and 3.38 16.63 (broad sAH,4-I-1), 6.70 (broad dlHj=s.s Hz,2'-H2), 7.12 (dIHJ=8.4 Hz,1'-I-D; IRv~~ (neat) 3270, 1730, 1615 cm- 1 MS m/e 523 508 142 (C2H 4

CON(CH

3 )0 4

H

9 47%).

-99- 16-HALO-ESTPJADIOL UNDECANAMID (SCHEME 9) N-n-butyl.N-methl-1 1 .17' B-diacetoy-,estra-l' 6'-tetraeng-zylI undecanamnide The ketone amtide 9b (163 mg, 0.50 ninol) was dissolved in isoprenyl acetate ml). p-toluenesulfonic acid (44 mg) was then added and the solution was distilled to about two-thirds of the original volume in 7 h and was then stirred *at reflux for 12 h. Afterwards, the solution was cooled with an ice-water bath and extracted with 50 ml of cooled ether. The ether was washed with a cooled satured sodium bicarbonate and water. The organic phase was dried with anhydrous MgSO 4 and the solvent was removed under reduced pressure. The residue was filtered through alumina (15mm. x 50 mm alumina Woehim neutral, activity ED) using a mixture of benzene-diethyl ether (3:7 v/vI) as eluant. The solvent was removed under reduced pressure and, the residue was purified by flash chromatography on silica gel. Elution with mixture of EtOAc/hexane (1:4 v/v) yielded the N-butyl, N-methyl-11-(3'17"0-diacetoxyestra-I',3',5'(1cy), 16'-tetraen-7'a-yl) undecanamidde (10) (244 mg, 80%) as colorless.

oil; IH-NM-R 8 (CDCI 3) 0.92 (s,3H-,18'-CH 3 0.92 and 0.95 (2t,3H-3',J=7.0

HZ,N(CH

2 3

CH

3 2.18 (s,3H,17'-OCOCH 3 2.28(s,3H'-OCOCH 3 2.76 app (d,lHJ16.1 Hz, part of ABX systemn,6'-H), 2.90 and 2.96 (2s,3H,N-CH 3 3.26 and 3.35 (2tapp,2H,J=7.6 Hz,N-CH 2 5.52 (mn,1H,16'-H), 6.80 (broad 6.85 (dd,1Hjj=9.1 Hz and J 2 =3.0 7.27 (d,1H,J=9.1 IR Vmax(neat) 100 1750, 1635, 1200 cml; MS m/e 607 5(M+-COCH 2 1 550

(M+-COCH

2

-CH

3 523 (M+-2COCH 2 142 (C2H 4

CON(CH

3

)C

4 H9+,55%), 129 (C4H9(CH 3 )NC0CH 3 114 (C4H9(CH 3 )NCO+, 86 (C4H 9

(CH

3 EXACT MASS calcd for C38H5705N 607.4239. found 607.4234.

N-butyl. N-mnethyl-i1(16'a-chloro-3'acetoxy-1 7-oxo-estra-1' 3'.41(1 0')-triene-7' cXyl) undecanamidde (11, X CD) To diacetate amide 10, dissolved in 5 ml of acetone, was added a solution of sodium acetate (2.6 equivalents) in acetic acid and water (1:11-3 v/v) and then, was treated with tertbutyl hypochlorite (1 eq.) prepared from t-butanol (4 ml) and javel water (Javex 50 nil). The dlear solution was warmed to 55*C and :stirred for 1 h. Afterwards, the solvent was evaporated to dryness. The residue was dissolved in ether (100 mld) and water was added (20 ml). The organic phase was washed with water, dried with anhydrous MgSO4 and evaporated to dryness. The residue was purified by chromatography on silica gel carried out with mixture of EtOAc/hexane (3:7 v/v) to give the N-butyl, N-methyl-Il- (16'a4-hloro-3'-acetoxy-l 7 '-oxo-estra-l',3',4'(1 0')-trien-7'a-yl) undecanamide 11, X--Cl) (115 mg, 89%) as colorless oil; IH-NMR S (CDC1 3 0.92 and 0.95 (2t,3H,J=7.0 Hz,N(CH 2 3

C-H

3 0.96 (s,3H,18'.-CH 3 2.28 (s,3H,3'-OCOCH 3 2.80 app (d,IH,J=16,6 Hz, part of ABX system, 2.90 and 2.96 (2s,3H,N-CH 3 3.24 and -101- 335 (2tapp,2H,J=7.4 Hz,-N-CH 2 4.46 (d,1H,j=6.6 Hz,16'P-H), 6.82 (broad 6.86 (dd,1H,J=9.1Hz and J 2 7.29 (d,1HJ=9.1 IR Vmax (neat 1750, 1640, 1205 cm- 1 MS m/e 601, 599 142

(C

2

H

4 CON(CH)C4H9 100%), 114 (C4H 9

(CH

3 )NCO 93%).

N-butylN-methyl- 1-(16'-chloro-3'.17'-dihydroxy-estra-l'.3' 5'(10')-trien-7'g-yl) undecanamide ("EM 139") and ("EM 170") A stirred solution of haloketone amide (11, X=CL) in anhydrous tetrahydrofuran (THF) (10 ml) under argon was chilled to -70 0 C with 2-propanol/dry ice bath. A solution of 1.0 M of lithium aluminium hybride (2 eq.) was then added dropwise. After 30 min, the reaction was allowed to return slowly at O*C for 5 min, then was quenched by the dropwise addition of a mixture of THF-EtOAc (5 ml) (1:1 v/v) and acidified at pH 4 with HC.

The mixture was stirring for 5 min at room temperature and then extracted with EtOAc. The organic phase was washed with water, dried on anhydrous Na 2

SO

4 and evaporated under reduced pressure. The residue was chromatographed on silica gel with a mixture of EtOAc/hexane (4:6 v/v) as eluant: -102- N-butyl, N-methyl-ll-(1 6'a-hloro-3' .17'c-dihydroxy..estra- O)-trier-7'aundecanamide ("EM 170") mg, 29%) as colorless oil; analytical sample was obtained by HPLC purification; IH-NMR 8 (CDC1 3 400 MI-z) 0.79 (s,3H,18'-CH 3 0.93 and 0.96 (2t, 3H,J=7.3 HzN(CH 2 3

CH

3 2.80 (2Hj6J=17.1 Hz and J 6 7 4.5 Hz, AS=24.34 (Hz, system ABX, 2-94 and 2.99 (2s, 3H9N-CH 3 3.26 (ddJ 1 7.6 Hz and J2 7.4 Hz) and 3.32-3.43 (m)-[2H-N-CH 2 3.71 (d,lHJ=4.5 Hz,17'J3-H), 4.63 (ddd, IH, J16115 10.2 Hz, J1, 1 7-=4.5 Hz and J 16 15 3.9 Hz, 16'0-H), 6.50 IH, J=24 Hz, 6.60 1HJ=2.5 Hz, 6.66 (dd,1HJ1=8 Hz and J 2 =2-5 Hz, 7.14 Hz, IRvna(neat) 3300, 1615. 1495 cnl; MS m/e 561,559 100%), 523 (M+-HC1, 142 (C2H4CON(CH 3

)C

4

H

9 114 (C4H9(CH 3 )CNO+, Exact mass calculated for C34H540 3

N

3 5 Cl 559.3785, found 559.3821.

-and -N-buyl. N-methyl-Il-(16'a-chloro-3'.l7'fdihydroxyestra 0')-trien-7Tayl) undecanamide ("EM 139') mg, 55%) as a colorless oil; analytical sample was obtained by HPLC purification; IH-NMR 5 (CDC 3 400 MHz), 0.81 (s,3H, 18'-CH 3 0.93 and 0.96 (2t, 3H,J=7.3 Hz, (CH 2 3

CH

3 2.78 2H, J6,6=16.2 Hz and J 6 ,7 4.5 Hz, A 5 =24.34 Hz, system AEX, 2.94 and 2.99 (2s, 3H,N=CH 3 327 (dd, JI-7.6 Hz and J 2 -7.5 Hz) 103 and 331-3.45 (in, 2H, -N-CH 2 ),3.86 (dd, 1H, J17,17-0H=3-4 Hz and J 17 ,1 6 =5.9 Hz, I~xHand 4.11 (ddd, 11-1 J 1 6,1 5 10.8 z J 16 17 5.9 z and J 16 15 =2-5 Hz, l6' 6.56 1H, J=19.7 Hiz, 6.61 1H, J=25 Hz, 6.66 (dd, 1H, J 1 =8.4 Hz and J 2 =2-6 Hz, 7.13 1H, J=8.4 Hz, ER vmax(neat) 3320, 1615.

1490 cml; MS m/e 561,559 38%, 100%), 523 (M+-HIi, 142 (C2H2CON(CH,3)C 4

H

9 114 (C4-1 9

(CH

3 )NCO+, exact mass calculated *0 for C34Hs4ON 3 5CI 5593785, found 559.382.

Scheme 9 0

OCOCH

3 HO **IfR

CH

*Il 41X 1 BA 13 X 9LRbKR EM* 17* A R= c EM16...1 tO c4 -105- N-n-butyl. N-methyl-l -(16'a-bromo-3'-acetox- 7'-oxo-estra-1' 3' trien-7'a-yl) undecanamide (11, X=Br) To the above diacetate 10 (244 mg, 0.40 mmol) dissolved in 10 min of acetic acid was added dropwise with stirring within 10 minutes and at room temperature, a brominating solution composed of 50 mg (0.6 mmol) of sodium acetate, 1.6 ml Sof acetic acid, 0.04 ml of water and 63.9 mg (0.02 ml, 0.40 mmol) of bromine.

During the course of this reaction, a red coloration appeared and disappeared.

To the solution, 50 ml of ether was added and the organic phase was washed with water (4 x 50 ml) followed by a saturated sodium bicarbonate solution (2 x ml) and finally with water (3 x 50 ml). The combined phase was dried over anhydrous magnesium sulfate and the solvent was removed in vacuo. The e residue was chromatographed on silica gel (Kieselgel, 60F254, Merck, *0.063-0.200 mm). Elution with a mixture of hexane-ethyl acetate (4:1 v/v) yielded N-butyl, N-methyl- 1-(16'a-bromo-3'-acetoxy-17' -oxo-estra-1',3',5'(10'), :trien-7'-a-yl) undecanamide (11, X=Br) (201 mg, as colorless oil; 1

H-NMR

8 (CDC1 3 0.94 3H,18'-CH3), 228 (s,3H, 3'-OCOCH 3 2.82 app (dlHJ=16.4 Hz, part of ABX system, 2.90 and 2.96 (2s, 3H,H-CH 3 3.24 and 3.35 (2tapp,2H, J=7.7 Hz, -N-CH 2 4.58 (t,1H,J=3.6 Hz, 16VH), 6.82 (broad 6.88 (dd,1H, J=8.0 Hz and J 2 =4.0 Hz, 7.29 (d,1H,J=8 Hz, MS m/e 644 565 Br, 522 (M+-Br-COCH 2 142 (C2H4CON(CH 3

)C

4

H

9 114 (C4H9(CH 3 )NCO 88 (100%).

-106- N-bu tyl. N-methyl-l l-(1 6'a-bromo-3'.1 7-dihydroxy-estra-1 (I O')-trien-7avi) undecanamide (nEM 105") and ("EM 171") A solution of bromoketone amide 11 (X=Br) (295 mg, 0.46 mmol) in anhydrous tetrahydrofuran (10 ml) under argon was chilled to -70 0 C and a solution of M of lithium aluminium hybride in ether (0.92 ml, 0.92 mnol) was added dropwise with rapid magnetic strirring. After 30 min, the reaction was quenched by the dropwise addition of a mixture of T-F-ethyl acetate (1:1 vlv) and acidified by 10% hydrochloric acid. The mixture was strirring for 5 min at room temperature and then extracted with ethyl acetate. The organic phase was washed with water, dried on anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The residue was purified by chromatography on silica gel Elution with a mixture of hexane-ethyl acetate (73 v/v) gave: **too: N-n-butvl. N-methyl-ll-(16'-brom-3'.17'a:dihdroW-:sta Tn-vDl undt-angmirla (7b4 I7") (63 21%) as colorless oil; IH-NMR 6 (DCD13,400 MHz) 0.81 3H,18'-CH), 0.93 and 0.96 (2t, 3H,J=7.3 Hz,N(CH2)3CH3), 2.79 (2HJ66=16.6 I-z, J6,=4.7 Hz, AS=24.34 Hz, system ABX,6'-M), 2.94 and 2.99-(2s3H,N-CH3), 3.27 (dd,HJ=7.7 Hz and J2=7.5 Fiz, -N CH2-), 3.31-3.44 3.66 (dd,1 HJ17 17-1.4 FIz, -107- J17,1 6 4,3 Hz, 170-H), 4.68 (dt,IH,J 6 17 =4,3 Hz, m, J 1 6,15=9.7H1Z,161-H), 6.60 (d, 1H,J=2.4 Hz, 6.65 (dd, 1HJ=8.5 Hz and J 2 =2 Hz, 7.14 (dIHKp=8.5 Hz, IRVmax (neat) 3300, 1615, 1495 an-i 1 MS mle 605,603 523 (M+-HBr, 142 (C 2

H

4

CON(CH

3

)C

4 Hq+, 100%), 114 (C4H9(CH 3 )NCO+, 97%); Exact mass calculated for C34H5403N7 9 Br 603.8289, found 603.3304.

and N-n-butyl. N-methyl-l -1-06'a-bromo-3'. 1Bdihydroy-estra-1 O)-tien- U* 7a-y) undecanamide ("EM 105-).

(170 mg, 50%) as a colorless oil; analytical sample was obtained by HFLC purification; IH-NMR 8 (CDC1 3 400 MHz), 0.80 3H,18,-CH 3 0.93 and 0.96 (2t,3HJ=7.3 HzN(CH 2 3

C-

3 2-80 (2HJ 61 6=164,J 6 7 =46 Hz, A&=24.34 Hz,*system ABX, 2.94 and 2,99 (2s,3HN-CH 3 3.27 (dd, 2HJ 1 =7.7 Hz and J 2 =7.5 Hz,

-N-C-

2 331-3.45 (m,2H,-N-CH 2 4.02 (ddIH 1 7 1 7 3.7 Hz, and J 1 7 16 =6.1 Hz, e. 17a-H), 4.15 (ddd,I1HJ 161 5 =10.2 Hz, J 16 17 =6.1 Hz and J16,5=2.9 Hz, 1613-1), 6.61 Hz, 6.66 (dd,lHJ=84 Hz and J 2 25 Hz, 7.12 (d,lHJ=8.4 Hz, IR v (neat) 3320, 1610, 1490 cn- 1 MS m/e 605, 603 523 (M+-HBr, 100%), 142 (C2H4CON(CH 3

)C

4

H

9 114 (C4H 9 (CH3)NCO, Exact mass calculated for C34H540 3

N

7 9 Br 6033289 found 6033289.

-108- N-butyl. N-methyl-i 1-1 6'a-iodo-3', 7lT-dihydroxy-estra-l '3'.-5'(10')-trien-7c- V1) undecanamide ("EM 156").

Under argon atmosphere, a mixture of 16a-bromodiol EM 105 (55 mg, 0.091 mmol) and dry sodium iodide (136 mg, 0.91 mmol) in freshly ethyl methyl ketone (25 ml) was refluxed in darkness during 12 hI Afterwards, the solvent was evaporated, water was added and the product was -extracted with ethyl acetate. The organic phase was washed with 5% sodium thiosulfate and with water, dried over anydrous sodium sulfate and concentrated to dryness under reduced pressure. The residue was purified by chromatography. Elution with a mixture of hexane-ethyl acetate v/v) gave a mixture of starting material and iodo compound (52-48) of which HPLC separation afforded N-butyl, N- .methyl-l1,(6'-a-iodo-3',17j-dihydroxy-estra-l'3'5'(10 -trien-7a-yl) undecananmide ("EM 156") (21 mg,36%) as colorless oil; IH-NMR 8 (CDC1 3 400 :MHz) 0.78 (s,3H,18'-CH 3 0.93 and 0.96 (2t,3H,J=73 Hz, N(CH 2 3

CH

3 2.79 (2H,J 6 6 =16.5 Hz,J 6 7 =4.4 HzAS, 2434 Hz, system ABX, 2.94 and 2,99 (2s,3H,N-CH 3 3.27 (dd,2H,J 1 -7.6 Hz and J 2 -7.5 Hz, -N-CH 2 3.32-3.44 2H

N-CH

2 4.09-4.17 2H, 16'PH and 17a-H), 6.60 (d,1H,J=2.4 Hz, 6.65 (dd,1H,J=8.4 Hz and J 2 -2.4 Hz, 7.13 (d,1H,J-8.4 Hz, IR v (neat) 3310, 1610, 1490 cm- 1 MS m/e 651 523 100%), 508 (M+-HI-CH 3 ,38%) 142 (C2H 4

CON(CH

3

)C

4

H

9 114 (C 4

H

9

(CH

3 )NCO*, Exact mass calculated for C 3 4H540 3 HI-HI 523.4028, found 523.4028.

-109- EFFICACY OF AN ANTIESTROGEN SYNTHESIZED IN ACCORDANCE WITH EXAMPLE 9 Compound "EM 139" shown in Scheme 9 above is an estrogen activity inhibitor. "EM 139" has been tested both for efficacy in acting as an antiestrogen by blocking estrogen receptors without substantially activating those receptors (see Figure and for efficacy in inhibiting 17-hydroxysteroid dehydrogenase '(see Figure an enzyme which catalyzes reactions involved in the synthesis of both androgens and estrogen (hereinafter referred to as "17f-HSD") The antiestrogenic activity of "EM 139" was measured as its ability to inhibit the estradiol-induced stimulation of uterine weight in adult female ovariectomized Balb/c mice (body weight=19-20 g) sacrificed five days after ovariectomy. "EM 139", and/or estradiol dissolved in ethanol were injected oo "..."subcutaneously in the appropriate groups in a solution of 0.9% sodium chloride and 1% gelatin at different concentrations of "EM 139" (as noted along the X axis of Figure A dosage of 0.2 ml of the foregoing preparation, was administered twice daily, starting on the day of ovariectomy for a total of 9 injections. Estradiol was injected at the dose of 0.01 ,g in 0.2 ml, twice daily, starting on the morning after ovariectomy for a total of 8 injections.

After sacrifice, the uteri were rapidly removed, freed from fat and connective tissue and weighed. Results shown in Figure 3 are the means SEM of groups -110of 9-10 mice. As may be seen from Figure 3, EM 139 was highly effective in reducing estradiol-induoed uterine weight gain.

To test the effect of "EM 139" on the inhibition of sex steroid formation, its effect on the 17p-hydroxysteroid dehydrogenase catalyzed conversion of estradiol to estrone was observed. The reaction was followed by monitoring formation of NADH (at 340 nm). The rate of conversion of cofactor NAD to NADH varies directly with the rate of estradiol conversion to estrone. The ability of "EM 139" to inhibit estrone production is indicative of its ability to inhibit the estrogen-forming reverse reaction because both reactions are catalyzed by 17p-hydroxysteroid dehydrogenase (Thomas et aL, J. BioL Chem.

258: 11500-11504, 1983). 17p-hydroxysteroid dehydrogenase (17f-HSD) was purified to homogeneity from human placenta. A reaction vessel was prepared containing 1 gg 17p-HSD, 5 mM NAD, 20 tM 17p-estradiol, and the concentrations of the test compound "EM 139" which are indicated along the *X-axis of Figure 4 in 1.0 ml of a mixture of Tris-HC1 (50 mM), EDTA (2 mM), NaN3 (5 mM). The pH was 7.5. The reaction was allowed to proceed at 25 0 C for min. Formation of NADH was measured at 340 nm. As shown by Figure 4, increasing concentrations of EM 139 significantly inhibited the reaction.

-111- EXAMPLE N-n-BUTYL, N-METHYL-I 1-(3',lT -DHYDROXY-I Ta-EHYNYL-F pRA- 15'-TETRAEN-Ta-YL) UNDECANAMIDE (CEM 123") (SCHEME N-n-butyl. N-methyl-ll-(3'-benzoyloxy-Ia-ethvlenediox estra-l',3'.5'-(10')- .trien-7 gz-vl) undecananide (12).

A mixture of N-n-butyl, N-methyl-I I-(3'-benzoyloxy-17-oxo estra-1',3',5'(10')trien-7-a-yl) undecanamide (9a) (3.63 ethylene glycol (215 ml), ptoluenesulfonic acid (530 mg) and anhydrous benzene (250 ml) was refluxed with a Dean-Stark apparatus during 24 h. After cooling, the mixture was poured in water and extracted three times with ether. The organic layer was washed with a saturated sodium bicarbonate solution, and brine dried on magnesium sulfate and evaporated to dryness. The residue was purified by flash chromatography on silica-gel (Kieselgel 60, Merck, 230 mesh ASTM, 300 g).

Elution with a mixture of hexane-ethyl acetate (6:4 vlv) gave pure N-butyl, N-methyl-ll-(3'-benzoyloxy-17'-ethylenedioxy estra-1,3'5'(10'), trien-7'-yl) undecanamide (3.58 g, 92%) as an oil, the structure of which was confirmed by spectroscopic means.

-112- N-ri-butyl. N-methyl-ll-(3'-benzovloxcy-16a-roo- 7-ehleedox-etr- ,trien-7a-yl) undecanamide (13 To the above ethylenedioxy amide 12 (370 mg, 0.55 mmol) in anhydrous tetrahydrofuran (10 ml) cooled at 0 4 'C was added dropwise under argon, a solution of pyridinium bromide perbromidde (406 mg, 1.36 nunol) in 7 mld of the same solvent. After stirring during 1.5 h at 0 0 C, sodium iodide (300 mg) was added and the solution was stirred for 25 min. Afterwards, a solution of sodium thiosulfate vlv, 10 ml) and pyridine (0.5 mld) was added and the mixture was stirred for an additional 4 h and then poured into water and extracted three times with ether. The organic layers were washed with 1N hydrochloric add, water, saturated bicarbonate solution and water dried on magnesium sulfate and evaporated to dryness. The residue was chromatographed on silica-gel (50 Elution with a mixture of hexane-ethyl acetate (4:1 v/v) gave pure N-n-butyl, N-methyl-1l(3'-benzoyloxy-16'a-bromo- 17'-ethylenedioxy-estra-l' trien-Tcx-yl) undecanamidde (13) (313 mg, 76%) as colorless oil; IR vma (neat), 1730, 1630, 1595 and 1255 cm 4 IH MM, 0.93 O3H, s, 18'-MH 3 ),2-28 (21-1, td, J 7-5 and 2-6 Hz, -CH 2 CON-), 2.90 and 2.95 (3H, 2s, -N-CH- 3 3.24 and 3.35 (2K, 2t, J 7.3 Hz, -N-CH 2 3.85 and 4.35 (4H, m,

-OCH

2 Ch 2 4-56 (1IH, m, 6.91 (IH, d, J 2.2 Hz, 6.98 (1IH, dd, J 8.4 and 2.2 Hz, 7.32 (1H, d, J 8.4 Hz, H-CV1), 7.49 (2H, tapp J= Hz H-C-3" and 7.63 (1H, tapp, J 7.0 H-z H-C.4" and 8.17 (2H, d, J -113- Hz, H-C.2" and MS m/e, 671 114 (C 4

H

9

(CH

3 NCO+, 13%), 105 (C6H 5 CO+, 100%), 86 (C 4

H

9

(CH

3 N~r 77 (C 6 1-1 5 +125%).

N-n-butyl, N-methyl-llI-(3'-hydroxvy-IT7-oxo-estra-1' 3' 5001) 1 5'-tetraen-7'nyl) undecanarnide "(EM 112)" To a solution of the bromoketal (13) (517 m&g 0.69 mmol) in anhydrous dimethyl sulfoxide warmed at 73*C, under argon, was ac!:1ed potassium-t-butoxide (1-55 g, 13.8 mmol). The midxture was stirred for 5 h at this temperature and then cooled, poured in ice-water, acidified with IN hydro-chioric acid and extracted three times with ethyl acetate. The organic :layers were washed with water dried on magnesium sulfate and evaporated to dryness. The residue was dissolved -in acetone (30 ml), water (7 ml) and p-toluenesulfonic acid (60 mg) was added. The resulting solution was stirred for 5 h at room temperature and then poured into water. The organic compound was extracted three times with ether, washed with a saturated sodium bicarbonate solution and water dried on magnesium sulfate and evaporated to dryness. The residue was purified by "flash chromatography" (100 Elution with a mixture of hexane-ethyl acetate (1:1 vv) gave the pure N-butyl, N-xnethyi.-1-(3-hydroxy-17'-oxo--estra-' 3' 15'-tetraen-7a-yl) undecanamide t1 EM 112" (178 mg, 49%) as colorless oil; IR Vmax (neat) 3290, -114- 1695, 1620 and 1600 cm-1; 1 H NMR, 0.92 and 0.95 (3H, 2t, J 73 and 7.0 Hz,

-N-(CH

2 3 CH3), 1.11 (3H, s, 18'-CH 3 2.32 (2H, td, J 2.5 and 7.0 Hz, 2.94 and 2.99 (3H, 2s, N-CH 3 3.27 and 3.38 (2H, 2t, J 7.7 and 73 Hz,-N-CH 2 6.11 (IH, dd, J 62 and 33 Hz, H-C15'), 6.66 (IH, d, J 26 Hz, H-C.4, 6.71 (1H, dd, J 8.4 and 2.6 Hz, H-C.2, 7.13 (1H, d, J 8.4 Hz, 7.60 (1H, dd, J 6.2 and Hz, H-C.16') and 7.70 (1H, broad s, w/2=16 Hz, OH), MS m/e, 521 53%), 507 (M+-CH 2 506 (M+-CH 3 142 (C2H 4 CON (CH3) C 4 H9+, 114 (C4H9(CH 3 )NCO+, 60%) and 86 (C 4

H

9

(CH

3 22%, 44 (100%).

.N-n-butyl, N-methyl-l B-dihydroxv-17-a-ethynyl-estra- tetraen-Ta-l) undecanamide ("EM-123") To hexanes (1 ml) cooled at 0C, were added trimethylsilylacetylene (0.112 ml), n-butyllithiuin 1.6 M in hexanes (0.25 ml), few drops of anhydrous THF and finally, a slowly addition of a solution of enone amide EM 112 (57 mg) in anhydrous THF (1.2 ml). The mixture was stirred for 30 min at 0*C. After addition of a saturated ammonium chloride solution, the mixture was extracted with ethyl acetate The organic layers were washed with water and brine, dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure. To the residue (61 mg) dissolved in methanol, a 5N potassium hydroxyde solution (0.177 ml) was added and the -115mixture refluxed for 50 min. After cooling and addition of a saturated ammonium chloride solution, the mixture was extracted three times with ethyl acetate. The organic layers were washed with brine, dried over magnesium sulfate and filtered. The organic solvent was removed under reduce pressure.

The residue was chromatographied on silica-gel (5 Elution with a mixture of hexanes: ethyl acetate (7:3 v/v) gave N-butyl, N-methyl-11-(3',17-dihydroxy- 1a-ethynyl-estra-l',3',5'-(10'), 1 5'-tetraen-7'a-yl)undecanamide ("EM-123") (34 mg, IR vmax (neat), 3290, 2150,1620 and 1600 cm- 1 H NMR, 0.92 and 0.95 (3H, 2t, J 7.3 and 7.0 Hz, N-(CH 2 3

C

3 0.95 (3H, s, 18'-CH 3 2.32 (2H, td, J and 2. Hz, -CH 2 CON-), 2.66 (1H, s, 2.93 and 2.98 (3H, 2s, N-CH 3 3.27 and 338 (2H, t, J 7.0 Hz, -N-CH1 2 5.78 (1IH, dd, j 5.9 and 33 Hz, 6.05 (IH, dd, j 5.9 and 1.5 Hz, H-C16), 6.62 (1H, d, J 2.5 Hz, 6.67 (1H, dd, J 8.4 and 2.6 Hz, H-C2') and 7.13 (1H, d, J 8.4 Hz, ppm; MS m/e 547 142 (C2H 4

CON(CH

3

)C

4

H

9 21%) 114 (C 4

H

9

(CH

3 )NCO, 88 (100%) and 86 (C4H 9

(CH

3 34%).

.9 9 116- EXAMPLE 11 I 6j3-CYCLOFROPYL

DERIVATIVES

(Scheme 11) N-n-bu X1, N-methyl-i i-(1 7-oxo-3'-hydroxy-1 5 1 0, 6 metnylene-estra l'-.S(lO'-trien-7rx-yl) umdecanamide (14) *A solution of the phenol-enone EM-112 (101 mg; 0.19 mmnol) dissolved in anhydrous pyridine (15 mal) and acetic anhydride (10 mld) was stirred at room temperature for 20 h. The mixture was into ice-water, then extracted three times with ether. The organic layers were washed with IN hydrochloric acid, water and a saturated sodium bicarbonate solution and water, dried on magnesium sulfate and evaporated to dryness. The residue was purified by "flash chromatography" on silica-gel (20 Elution with a mixture of sees9 hexane-ethyl acetate (73 v/v) gave the N-butyl, N-methyl-11-(17'-oxo-3'ace toxy-es tra-I 5'-tetraen-7a-yl) undecanamide.

99999. To this and palladium (OD1 acetate (11 mg) in ether (25 ml) an ethereal diazomethane solution (prepared from I g of diazald) was added dropwise at 0 0 C with continuous stirring during 10 min. After evaporation, the residue was dissolved in methanol (50 mld) and cooled to 2N sodium hydroxyde solution (imi) was added and after 75 min. of stirring the mixture was neutralized with IN hydrochloric acid, extracted three times with ether. The organic layers were washed with brine, dried on magnesium sulfate and -117evaporated to dryness. The residue was purified by HPLC to give N-butyl, Nmethyl-l -(17-oxo-3'-hydroxy-15'p, 16'-methylene-estra-1',3',5'(10')-trien-7ayl) undecanamide (14) (79 mg, 76%) as a colorless oil. IR vmax (neat) 3260, 1705, 1610 and 1570 ac-1; 1 H NMR (400 MHz) 0.93 and 0.96 (3H, 2t, J 73 Hz,

N(CH

2 3

C-H

3 0.99 (3H, s, 18'-CH1 3 1.98 (1H, td, J= 83 and 3.96 Hz, H-C16'), 2.80 (IH, d, J 16.6 Hz, 2.94 and 2.98 (3H, 2s, N-CH3), 3.27 (1H, dd, J 7.58 and 6.66 Hz) and 3.38 (1H, m) (both are -N-CH 2 6.64 (1H, d, J 2.6 Hz, 6.66 (IH, dd, J 8.2 and 2.6 Hz, and 7.10 (1H, d, J 8.2 Hz, H-Cl') ppm; MS m/e 535 522 (M+-CH2, 129 (C4H9(CH 3

)NCOCH

3 114 (C4H9(CH)NCO+, 67% and 88 (100%).

N-n-butyl N-methyl-l 17B-dihydroxy-15'B,16'B-methylene-estra- 3'5'(1 0')-trien-7n-yl) undecanamide (EM-136) To the cydclopropylketone 14 (10 mg, 18.7 ;mol) dissolved in methanol (8 ml) was added sodium borohydride (1.5 mg). The mixture was stirred at room temperature for 18 h. After addition of water, the mixture was concentrated under reduced pressure. The residue was diluted with water and extracted three times with ethylacetate. The organic layers were washed with brine, dried over magnesium sulfate and filtered. The organic solvent was removed under reduced pressure and the residue was purified by "flash chromatography" on -118silica-gel (5 Elution with a mixture of hexanes: ethyl acetate (5:5 v/v) gave N-butylN-methyl-1 7'-dihydroxy-15',16'P-cyclopropyl-estra-l',3,5(10)trien-7'ci-yl) undecanamide as a colorless oil, IR vmax(neat) 3300,1615,1580 and 1450 cm-1, H NMR (400 M4, 0.31 (IH, dd, j 14.0 and 7.8 Hz, H-C1") 0.53 (3H, s, 18'-CH 3 0.93 and 0.96 2t, J 7.3 Hz, N(CH 2 3 Cj3), 2.7 d, J 17.1 Hz, H1-C6), 294 and 2.98 2s, N-CH 3 3.27 (1H, dd, J 7.7 and 7.5 Hz) and 3-39 (1H, n) (both are -N-CH 2 4.09 (II- broad s, w 10 Hz, H-C17), 6.64(2, m, H-C4' and H-C-2' and 7.11 d, J 8.3 Hz, ppm; MS i/e 537 519 (M+-H 2 O, 504 (M+-H 2

O-CH

3 100%), 142 (C2H 4 CON(CH3)C4H 9 114 (C4H9(CH 3 )NCO+, 50%) and 86 (C4H 9

(CH

3 33%).

N-n-butyl, N-methyl-I 7'-dihvdroxy-17 -ethynvl-158.1 6'-methvleneestra-I'3',5'(10')-trien-Tpvl) undecananide ("EM-138") To hexanes (500 cooled at 0C, were added trimethylsilylacetylene (54.6 g), 1.6 M n-butyl lithium in hexanes (120.4 id), few drops of anhydrous TIF and finally, a slowly addition of a solution of the cyclopropyl ketone 14 (25.8 mg) in anhydrous THF (350 iii). The mixture was stirred for 75 min at OOC. After addition of a saturated ammonium chloride solution (1 ml), the mixture was extracted three times with ethyl acetate. The organic layers were washed and -119brine, dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure. To the residue dissolved in methanol (900 PI), a potassium hydroxyde solution (70 p1) was added and the mixture refluxed for min. After cooling and addition of a saturated ammonium chloride solution (1 ml), the mixture was extracted three times with ethyl acetate. The organic layers were with washed with brine, dried over magnesium sulfate and filtered.

The organic solvent was removed under reduced pressure. The residue was purified by "flash chromatography" on silica-gel (5 Elution with a mixture of hexanes: ethyl acetate (55 v/v) gave N-butyl, N-methyl-11-(3',17-dihydroxy- .Ta-ethynyl-15'P,16 1 -cyclopropyl-estra-1,3',5'(10')-trien-Ta-yl) undecanamide ("EM 138") (12 mg, 44%) as a colorless oil; IR vmax (neat) 3590, 3300, 1620, 1600 and 1450 cm-1; 1 H NMR (400 MHz), 039 (1H, ddd, J 14.6 and 7.9 Hz, 0.93 and 0.96 (3H, 2t, J 7.4 and 73 Hz, -N(CH 2 3

-CH_

3 0.96 (3H, s, 18'-CH 3 2.70 S(1H, s, -C CH), 277 H, d, J 165 Hz, 2.94 and 2.98 (3H, s, N-CH 3 3.27 (1H, dd, J 7.7 and 7.6 Hz) and 3.38 (1H, m) (both are N-CH 2 6.42 (1H, m, OH), 6.65 (2H, m, H-C4' and and 7.12 (1H, d, J 83 Hz, ppm; MS m/e 561 142 (C2H4CON(CH 3 )C4H 9 114 (C 4

H

9

(CH

3 88 (100%) and 86 (C4H9(CH 3 120 Scheme I1I H

H

H H CHC2(C jCO(H)4

C

6

H

5 C0 2 'C 2 1

CNC

3

CH

"(CHJ)

,CON(CH

3

)C

4

H

9 H HO

'J(CH

210 C0N(CH 3

)C

4

HJ

Em 112 EM 123 121 EM 112 a.

9 a "(CU oCON(CUt)C 4

HI

"-(CH

2 10 CON(CU4)C 4 fn

"(CH-

2 10 C0N(CH- 3

)C

4 N1 Em. 136 Em 138 122- EXAMPLE 12 17a-ALKYNYLAMIDE

ESTRADIOLS

GENERAL PROCEDURE FOR ESTER FORMATION (SCHEMA 12) In anhydrous conditions under argon atmosphere, bromo acid (17 mmol) was dissolved in dry CH 2 C1 2 (30 ml), oxalyl chloride (12 ml) was added and the reaction was stirred 2 h at room temperature. Then, dry benzene was added to mixture and solvent was evaporated under reduced pressure (2 x) and dried under vaccum. This crude product was dissolved in dry CH2C1 2 (10 ml) and added at 0°C to a solutin of 3-methyl 3-oxetanemethanol (17 mmol), CH 2 C12, (7 ml) and pyridine (1.4 ml). The reaction was kept at this temperature for 4-8 h.

Thereafter, mixture was diluted with CH 2 C1 2 washed with NaHCO 3 v/w) and organic phase was dried over MgSO4. After removal of solvent, residue was purified by chromatography (hexane-ethyl acetate-triethylamine 80:20:1, v/v/v) to afford bromo ester.

6-bromo hexanoate ester of 3-methvl-3-hydroxymethyloxetane Light yellow liquid (91% yield); IR v (neat) 2930, 2860, 1725 1450, 1370, 1160 cm- 1 5 (CDC13) 131 3H), 1.1-2.1 6H), 2.36 J 6.0 Hz, 2H), 3.36 J 6 Hz, 2H), 4.13 2H), 4.41 (AB system av 83, J 6 Hz, 4H).

9-bromo nonanoate ester of 3-methyl-3-hydroxymnethyl oxetane (16).

Colorless liquid (86% yield); IR V (neat) 2920, 2840, 1725, 1450, 1370, 1150 cm 4 1; 5 (CDC1 3 1.31 11H), 1.2-2.2 (in, 41-1), 2.40 6.0 Hz, 2.HD, 3.45 J HIz, 2H), 4-20 21Df, 4.20 2MH, 4.48 (AB system av 8.2, J 6.0 H-z, 4H-).

1-bromo undecanoate ester of 3-methyl-3-hydroxymethyl oxetane (17).

Colorless liquid (85% yield); NMR-60 5 (CDC1 3 1-33 1511), 1.0-20 (in, 41-1), 2.30 J 6.0 Hz, 21-fl, 3.35 J 6.0 Hz, 211), 4.12 4.40 (AB system,& J ~6.0 H-z, 4W-f.

I 2-bromo dodecanoate ester of 3-methyl-3-hydroxymethyl oxetane 1,18).

Colorless liquid (86% yield): IR v (neat) 2910, 2840, 1720, 1450, 1370, 1155 cm 4 1; 8 (CDCl 3 1.30 17H), 1.1-2-0 (mn, 41-f, 2-30 J 6.0 Hz, 21-),3-33 J Hz, 211), 4.11 2H), 4.40 (AB system A v 8.0, J 6.0 H, 4M-.

-124- GENERAL PROCEDURE FOR ORTHO ESTER FORMATION (SCHEME 12) To a solution of bromo ester (3.4-14-2 mmol) in dry CH 2

CI

2 (100 mi) at OOC was added with stirring distilled boron trifluoride etherate (0.85-3.55 mmol).

After 4 h at OOC, reaction mixture was quenched by the addition of triethylamine (3.4-14.2 mol), diluted with diethylether and filtered to remove the amnine-BF 3 complex. The filtrate was evaporated and residue was purified by chromatography (hexane-ethylacetate-triethylamine 8020:1, v/v/v) to give bromo ortho ester.

1-(5'-bromo pentanyl)4-methyl-2,6.7-trioxabicydo 12.221 octane (19).

Colorless oil (68% yield); IR v (neat) 2940, 2915, 2855, 1450, 1390, 1050, 980 cm 4 8 (CDC1 3 0.79 3H), 1.2-2.0 8H), 335 J 6.0 Hz, 2H), 3.87 6H); MS m/e (rel. intensity) 280 278 250 248 197 195 179 177 72 69 (100).

1-(8'-bromo octanl)-4-methyl-2,6,7-trioxabicydo f22.21 octane Amorphous white solid (69% yield); IR v (KBr) 2940, 2900, 2840, 1450, 1390, 1045, 985, 950 man- 1 NMR-60 6 (CDC1 3 0.80 3H), 133 8H), 1.0-2.1 6H), 3.40 J 6.0 Hz, 2H), 3.93 6H); MS m/e (rel. intensity) 323 321 292 290 239 237 221 219 69 55 (100).

0'-bromo decanyl)4-methyl-26,7-trioxabicyclo 12.2.21 octane (21).

White solid (74% yield); m.p. 51-53 0 C; IR v (KBr) 2940, 2900, 2850, 2830, 1455, 1390, 1055, 985, 955 can- 1 NMR-60 6 (CDC1 3 0.80 3H), 1.27 121H) 1.1-2.1 (m, 6H), 339 (t J 6.0 Hz, 2H11), 3.87 6H); MS m/e (rel. intensity) 350 348 321 319 269 248 144 (37) 55 (100).

126- 1 -0 Ebromo undecanallyl)-4-methyl.6,7trioxabiclo f 2,2,21 octane (22).

White solid yield); m-p. 47-5, 48-5(C; IRV M Kr) 2900, 2835, 1460, 1045, 975 cm-I; NM-60 6 (CDCi 3 0.79 3W, 1.25 14K-1, 1.1-2-1 3.37 J Hz, 2M, 3.85 6W; MS nile (rel. intensity) 364 Ulf, 362 334 (13), 332 283 263 (85),2,61 144 55 (100).

4. Preparation of 17ci-alkynylamide estradiols (scheme 13) 127 GENERAL PROCECURE FOR COUPLING REACTION (Scheme 13) In a flame-dried flask under argon atmosphere, 3,17p-bis tetrahydropyranyl ethynylestradiol 23 (1.5 mmol) synthesized from commercial ethynyl estradiol and dihydropyran was dissolved in dry THF (40 ml) and HMPA (6.0 mmol).

The solution was cooled at -78*C and n-BuLi (3.0 mmol) was added. After 2 h, appropriate bromo ortho ester 19-22 (6.0 mmol) in dry THF (10 ml) was added at -78'C. The mixture was allowed to return slowly at room temperature and kept at this temperature overnight Then, brine was added and the reaction mixture *was extracted with ethylacetate. The organic phase was dried over MgSO4 and the solvent was removed under reduced pressure. The crude product was purified by chromatography (hexane-ethylacetate-triethylamine 96:4:1 to 5050:1, v/v/v) to give coupling product 24-27, unreacted steroid 23 (61, 62, 57%) and small quantity of undeterminated product.

1-3',178-bis f(tetrahydro-2"H-pyran-2"yl)oy) estra-1'3',5'(10')-trien-T17a-yll-7- 4 -methyl-2',6',7-trioxabicyclo f2'.2'.2'1 octan-1'-yl)- -heptyne (24).

Colorless oil (15% yield); IR v (neat) 2920, 2855, 2230, w, 1600, 1485 cm- 1 8 (CDC1 3 0.75 3H), 0.88 3 2.80 2H), 3.2-4.1 4H), 3.80 6H), 4.9-5.3 -128- 5.34 1H), 6.75 2H), 7.19 J 8.0 Hz, 1H); MS m/E'(rel. intensity) 579 (M+-DHP, 564 494 477 374 85 (1000.

1-(3'178-bis f(tetrahydro-2"H-pyran-2"yl)oyl estra-1'3'5',(10')-trien-17-yli (4'-methyl-2'.6'.7-trioxabicylo octan-1'-ylO-1-dcne Colorless oil (15% yield); IR v (neat) 2915, 2850, 2210 w, 1600, 1485 cm- 1 NMR-200 8 (CDC1 3 0.79 3W), 0.90 31-1), 224 J 6.6 Hz, 211), 283 2H), 3.55 3.89 6H), 3.95 2H), 4.98 and 5.19 (2s, IN), 5-39 1H), 6.78 J =26Hz, IN), 6.84 (dd, J1 26 Hz and J 2 =8.4Hz, 11, 7.22 J 8.4hZ, 11-D; MS W/e (rel. intensity) 620 535 518 85 (100).

1-3'.178-bis I(tetrahydro-2"H-pyran-2y)oxyI estra-V.3'5',(10')-trien-17a-l)-12- (4'-methyl-2'.6',T-trioxabicydo 12'.2'.1 octan-1 '-yl)-2-dodecyne (26).

Colorless visquous oil (42% yield); IR v (neat) 2920, 2850, 2210 vw, 1600, 1485 cm 1 NMR-200 8 (CDC1 3 0.79 3H), 0.90 3H), 225 J 6.6 Hz, 2Mf, 283 (m, 2H), 3.55 21-1), 3.89 6K-, 3.95 2Mf, 5.0 and 5.2 (2s, 11f, 5.39 IN), 6.78 J 26 Hz, 1H), 6.84 (dd, J1 2.6 and J 2 8.4 Hz, 1H), 7.21 J 8.4 Hz, IH); MS m/e (rel. intensity) 649 (M+-DHP, 634 564 547 85 (100).

-129- 1 -f3'.1-bisf(tetrahydro-2"Hpyan-2"y1)oxyloestra-1 3'5(1 0)-trien-1 -1 7-yll-13- 4 methyl-2'.6'.7-trioxabigcdof2'.2'loctan-l'-yl)--tride cne (27) Colorless visquous oil (35% yield); ER v (neat) 2915, 2850, 2290 vw, 1600, 1490 cn- 1 NMR-200 S (CDCI 3 0.80 0.90 2.25 J 6.6 Hz, 2H), 283 (m, 3-53 2W, 3.89 61), 3.95 211), 5.0 and 52 (2s, 11), 5.39 1W), 6.78 J 2.2 Hz, 11), 6.84 (dd, J 1 26 and J 2 =8.4 Hz, 7.21 J =8.4 Hz, 1).

see* to. 000.

*000 0: 130 GENERAL PROCEDURE FOR ORTH-O ESTER AND DI-THP HYDROLYSIS The product with ortho ester and di-THIP group (0-22-0.63 mmol) was dissolved in MeOH (80-120 ml) and P-toluenesulfonic acid (0.17-0.23 mniol) was added.

The solution was stirred at room temperature for 2-3 IL Then, water was added, MeOH was removed under reduced pressure and residue was extracted with ethylacetate. After evaporation of solvent, the crude product was purified by column chromatography (hexane-ethylacetate 5;5, vlv) to give ester compound with free hydroxyl group.

8-(3%.17B-dihydroxy estra-lV.3,5'CIO')-trien-17a-yl)-7-octyoate ester of 2'2dihydroxyMethyl propanol (28).

Colorless visquous oil (70% yield); IR v (film) 3340, 2910, 2850, 1710, 1600, 1485 cm- 1 NMR-200 8 (CDCI 3 0.83 0.86 31-D), 2.27 J 6.4 Hz, 21-1), 2.38 (t, J 7.1 Hz, 2H), 2.81 (mn, 2H), 3-54 (s broad, 4H), 4.17 21-1), 4.87 1W1, 6.56 (d, J 2.6 Hz, 1H), 6.63 (dd, J 1 2-6 Hz and J 2 8.4 Hz, 1HM, 7.17 J 8.4 Hz, 1H); MS rn/e (rel. intensity) 512 14), 494 479 466 270 159 (100).

-131- 1I-(3'.1TB-dihroUy estra-1'3'5'(1O')-trien-LTn-vl)-10-undecvnoate ester of 2%2'dihydroxymethyl propanol (29).

Colorless visquous oil (61% yield); iR v (neat) 3360, 2910, 2840, 2210 vw, 1710, 1600, 1485 cm 1 NMR-200 (CDCl 3 8 0.84 3W), 0.86 311), 2.24 J 7.0 Hz, 2.79 21), 3.34 (s broad, 2H), 3.56 (s broad, 4M), 4.13 21-D, 6.57 (Sapp, 111), 6.63 (dd, J 1 2.6 Hz and J 2 8.4 Hz, 7.14 J= 8.4 Hz, 11) MS m/e (rel.

intensity) 554 536 520 507 435 419 270 160 133 (100).

gee& 17-dihydro estra-1 3'5*(10')-trien-1 7a-vl)-12-tridemoate ester of 2%2'-dihydrox!vmethyl pro~anol Colorless visquous oil (78% yield); IR v (film) 3360, 2915, 2840, 1710, 1600, 1490 cm- 1 NMR-200 6 (CDCI 3 083 64, 2-25 4H), 2.78 211), 3-53 (s broad, 4H), 4.09 21), 6.6 21-1), 7.10 J 8.0 Hz, 11-1); MS m/e (rel. intensity) 582 563 548 535 463 446 270 160 133 (58), (100).

132 1 4433 17'8-dihydroxy estra-1 ',3'S5(10O)-trien-lTaz-yl)-1 3-tetradecynoate ester of 2'.2T-dihydroxy]Methyl propanol (31).

Colorless visquous oil yield); IR v (film) 3360, 2910, 2840, 2220 vw, 1710, 1605, 1490 an- 1 NM{R-200 S (CDC1 3 0.85 0.87 3W1, 2.25 J 6.6 Hz, 21HD, 2-33 J=7.1 Hz, 2W, 2.80 (mn, 2W1, 2.9 (in, 2W-1, 3.58 (s broad, 41W, 4.20 (s, 2M-1, 5-72 111, 6.56 J= 2.6 Hz, 111), 6.62 (ddl, ji Hz and J 2 =8.4 Hz, 111, 7.15 (d,J 8.8 Hz, 111.

.00.0 133 GENERAL PROCEDURE FOR HYDROLYSIS OF ESTER FOLLOWING BY AMIDE

FORMATION

At a solution of ester (0.14-0.49 mmol) in MeOH (12-50 nml) was added aqueous solution of KOH 10% w/v (6-25 ml) and mixture was refluxed under argon atmosphere for 24 Thereafter, water was added and MeOH was evaporated.

under reduced pressure. The resulting solution was acidified with HCl and extracted with ethylacetate. Organic phase was washed with water, brine and dried over MgSO4. Without purification, the crude carboxylic acid (IR acid band at 1700 and 2400-3600 cm- 1 was dissolved in dry CH2CI 2 (20-70 ml) and tributylamine (0.58-2-.4 minol). The mixture was cooled at -10C, isobutyl chioroformate (0.68-2-41 mmol) was added and allowed to react 30 min. At this :time, N-methylbutylamidne in excess (4.2-16.0 mmuol) was added and the cooling bath was removed. After 2 h, CH 2 Cl 2 was added and organic phase was washed with HCl (IN) and dried over MgSO 4 The solvent was removed and crude amide purified by column chromatography (hexane-ethylacetate/7:3, v/v).

-134- N-bu tyl N-methl--3'-(i-bu tyloxy carbonloxv)-17B-hvdroy estra-1V-3 5'(10Q)trien-lY'a-ll-7-octynami (32).

Colorless oil (79% yield); IR v (neat) 3380,2920, 2850, 1745,1620 cm 4 l; NMR-200 6

(CDC

3 0.87 311), 0.91 and 0.94 (2t, J 73 Hz, 3H), 1.00 (di J 6.6 Hz, 6H), 2.85 (in, 2M), 2-89 and 2.91 (2s, 3H), 3.22 and 3.33 (2t J 7-5 Hz,2M), 4.02.(d, J 7.0 Hz, 2H), 6.88 J 2.6 Hz, 1H), 6.93 (dc, J 1 2-6 Hz and J 2 =8.4 Hz, 7.29 J 8.4 Hz, 1H); MS m/e (rel. intensity) 579 12), 561 546 461 447 270 57(100). EMS M+ calculated for C36H.30 5 N: 579.3923; found: 5793970.

N-butyl, N-methyl-l1-f3'-(i-bu yloy carbonloxv-1R&-hvdroxy estra- 13 5'(1 0')-trien-1'n-vll-1 0-undecynaninde (33).

Colorless oil (67% yield); JR v (neat) 3370, 2910, 2840, 1745, 1620 cm 4 NMR-200 8 (ODC 3 0.87 3H), 0.92 and 0.95 (2t, J 6.6 Hz, 3H), 1.00 J 7.0 Hz, 6H), 2.86 2H), 2.90 and 2.94 (2s, 3H), 3.24 and 3.35 (2t, J 7-3 Hz, 21), 4.03 J 6.6 Hz, 2H), 6.88 J 26 Hz. 1H), 6.93 (dd, J 1 2.6 Hz and J 2 8.8 Hz, 11-0, 7.30 J 8.1 Hz, IH); MS m/e (rel. intensity) 621 606 602 212 159 142 114 (100).

-135- N-buyl, N-methyl-i3-[3 (i-butyloxy carbonyloxv)-178-hydroxy estra-13 0)trien-1 7 k-vlI-12-tridecynamide (34).

Colorless oil (89% yield); IR v (neat) 3370,2920,2840, 1745, 1620 cur 1 NMR-200 S

(CDC

3 0.87 3Mf, 0.92 and 0.95 (2t, J 7.0 Hz, 1.00 J 7.0 Hz, 61-f, 2.86 2H), 2-90 and 2-96 (2s, 3M-f, 3.25 and 3.35 (2t. J 7.4 Hz, 2M, 4.02 J 6.6 Hz, 2H), 6.88 I 2.2 Hz, I1-1), 6.93 (dd, J 1 2.6 Hz and jj 8.4 Hz, 1K, 7.30 J 8.8 Hz, 1Kf; MS i/e (reL intensity) 649 20), 633 631 616 531 516 270 57 (100); EMS M+ calculated for C4 1 H630 5 N: 649.4706; found: 649.4643.

N-butyl, N-methl-14-3'(i-butloqy carbonloxv)-17'-hydroxy estra-1'3' trien-17acyll-13-tetradecnamide Colorless oil (83% yield); IR v (neat) 3380,2910,2840, 1750, 1625 airl; NMR-200 8

(CDCI

3 0.87 31f, 0.92 and 0.95 (2t, J 7.0 Hz, 3H), 1.00 J 6.6 Hz, 6Kf, 2.85 2H), 2.91 and 2.96 (2s, 3H), 3.25 and 3.36 (2t, J 7.4 Hz, 2H), 4.03 J 6.6 Hz, 210, 6.88 J 26 Hz, IH), 6.93 (dd, ji 29 Hz and J2 8.4Hz, I K, 730 J 8.8 Hz,IH).

-136- HYDROLYSIS OF CARBONATE Hydrolysis of carbonate compounds 32-35 was performed as follows: carbonate derivatives were dissolved in methanol (10 ml). K 2 CO3 p/v) in aqueous methanol (25:75, v/v) (10 ml) was added and the resulting solution was stirred at room temperature for 3 h. Reaction mixture was acidified with HCI (IN) and MeOH was evaporated under vacuum. The residue was extracted with ethyl acetate and organic phase was dried, evaporated and purified by column chromatography (hexane-ethyl acetate 6.5:3.5, v/v).

N-butyl. N-methyl-8-[3',17B-dihvdroxy estra-l',3',5'(10')-trien-17n-yl]-7octynamide ("EM 157").

Purified by column chromatography (hexane-ethyl acetate/4:6, v/v).

a.

Amorphous white solid (88% yield); IR v (film) 3280, 2910, 2840, 1610 cm-1; NMR-200 8 (CDCl 3 0.87 3H), 0.91 and 0.94 (2t, J 7.0 Hz, 3H), 2.80 2), 2.90 and 2.92 (2s, 3H), 3.22 and 3-34 (2t, J 73 Hz, 3H), 5.22 1H), 6.57 J 2.9 Hz, 1H), 6.64 (dd, Ji 2.6 Hz and J 2 8.4 Hz, 1H), 7.16 J 8.4 Hz, 1H); MS m/e (rel. intensity) 479 (M 11), 462 460 446 270 114 88 44 (100); EMS M calculated for C 31

H

45 0 3 N: 479.3399; found 479.3369.

-137- N-butvl, N-methyl- I-13'.178-dihydroxy estra-i'3'5'(10')-trien-17 undecynamide ("EM 183").

Purified by column chromatography (hexane-ethylacetate/4:6, v/v).

Amorphous white solid (83% yield); IR v (KBr) 3300, 2910, 2840, 1610 cm-1; NMR-200 (CDCl 3 0.87 3H), 0.93 and 0.95 (2t, J 7.0 Hz, 3H), 2.80 2H), 2.91 and 2.94 (2s, 3H), 3.23 and 335 (2t, J 73 Hz, 2H), 5.30 1H), 6.57 J 2.6 Hz, 11-1), 6.64 (dd, J 1 2.6 Hz and J 2 8.4 Hz, 1H), 7.16 J 8.1 Hz, MS m/e (rel. intensity) 521 505 502 489 487 270 114 88 44 (100).

N-bu tyl. N-methyl-1 34[3',1 78-dihydroxy estra-1l'3'5(1 0')-trien-1Th-yll-12tridecynamide ("EM 163").

Purified by column chromatography (hexane-ethylacetate/7:3, v/v).

Amorphous white solid (98% yield); IR v (film) 3300, 2910, 2840, 1610 cm- 1 NMR-200 8 (CDCl 3 0.88 3H), 0.93 and 0.95 (2t, J 7.0 Hz, 3H), 2.80 2H), 2.93 and 2.97 (2s, 3H), 3.25 and 338 (2t, J 7.5 Hz, 2H), 6.61 J 2.6 Hz, 1H), 6.69 (dd, J 1 2.6 Hz and J 2 8.6 Hz, 1M), 6.87 1H), 7.14 J 8.1 Hz, 1H); MS m/e (rel. intensity) 549.(M+, 532 530 516 270 114 88 44 (100); EMS M+ calculated for C36H 5 50 3 N: 549.4182, found: 549.4271.

-138 N-butyl. N-methyl-i 43%ITBD-dihydroxy estra-l-3S-(l 0')-trien-17'a-vyll-13tetradecvnamide ("EM 196").

Purified by column chromatography (hexane-ethyl acetate/6:4, vlv).

Amorphous white solid (93% yield); rR v (film) 3280, 2915, 2840, 1615 cm 4 NMR-200 S (CDCl 3 0.88 3M1, 0.94 and 0.95 (2t, J 7.0 Hz, 311, 2.80 (in, 21), 2.95 and 2.98 (2s, 3M-1, 3.26 and 339 (2t, J 73 Hz, 21-D), 6.61 J 2.2 Hz, 111), 6.70 (dd, JI 2.6 Hz and J2 =8.4 Hiz, 1M-1, 7.13. (mn, 211: aromatic and phenolic hydrogen).

139 Scheme 12 84Qc4 2 ),,COOH B4CH 2 ),.CO~c 2 Br(H C8 CH O o 0a a a a a. a a.

a.

a a 15. 1B 19-22 8. 10. 11) 8, 10, 11) (LL L 8*s -U) (Z(,,&0)VHO)OZHOOOO lc) z LL-u 96LK3 9-u -CBLVG s-urr sc 0o

HO*

)d~l t2.& 0.

dMS CL0 (6 V0 LVS-U) IZ V -014'l- 141 EXAMPLE 13 Scheme 14 N-n butyl, N-methyl-3(3-17p-dihydroxy-1 I p-methoxy estra l*,3',5(10O)-trien 7cc-yl) undecanamide ("EM 111V) and its 17a-ethynyl derivatives ("EM 121").

O 0 0 0

O

.H'

36 1 IA-H37 2) (HC~q,2O 0 0~ AcZO *38 39 MgBi(C8 2 )l 1 ThP QAc

OTBOMS

0 0 1) H 1) DIBAL-H 2)TBDMSCa.6 j 2) 0 "(CH 2 11 OTHP 0 "C2,OBM 142 *fl.

0 *00 0 0*00 0 0000 00 0 00 00 0 000000 0

OTBDMS

HO

DD 00 1 OTBDMS ox (C 6 1- 5 2

CH

2 (IC 6

HSI)

2

OTBDMS

HO

11 OH TBDMSOa 1) Na-I 2) CH 3

I

2 11

OTBDMS

OTBDMS

"(CH

2 11

OTBDMS

OTBDMS

Bu NF

TBDMSO"(CH

2 11 OTBDMS HO 143- 1) =0C 2 R, rBU 3

N

2) D~iC 4 tig 9@ 0*

S

1) NaOH 2) OfH "(CHWj 10

CONC

4

HN

EM ill I

CH

3 1) NaOH 2) HC--C-S(CH:) 3 nBuLi 3) KOH

OH

NaBH4 EM 121 1

CHM

3 -144- EXAMvfLE 14 Scheme 11 p-chloromethyl derivatives **0 N-n-butyl, N-methyl, (11 p-chloromethyl-3,17JP-dihydroxy-,estra-l trien-7kr-yl) undecanarnide (56) and its 17ci-ethynyl derivative (58) 1 OTBMS TBDMSO

S*

0 950 0 @0S4 O0SS 0. 0 90

OS

0 506055

S

145 f 1) Bu44NF 2) CVH 3 0OCA 'Jones' 11

OTBDMS

CH

3 COj CH3COY

CFI

3 ccv 2 )j 0 C0NC 4

H.

CH3CO2- 1) ct 2)H' "'(Cf 2

)IQCONC

4

H

9

CII

r J n a (A J 2) KOH

(CH

2 10

CONC

4 HIj

"'(CH

210

CONC

4 Hg 147 EXAMPLE Scheme 16 Compounds with aliphatic side-chain in l 7 1-position N-n-butyl, n-methyl, I 1-(17f -hydroxy-T-mnethoxy estra-1 3 ',F(1O')-trien- 1 7 czcz-yl) undecanamidde ("EM 1O3").

Cty" 1

OTWP

ROO RO,0 63 R-&LCtj% 64 O0

RO'I

2) COjf*C EM 103 -148- EXAMPLE 16 Scheme 17 1 7 a-cyclopropyl derivatives N-n-butyl, N-methyl-(17a-cyclopropyl-3,173-dihydroxy estra-1 ,3',5(10')-trien-

T

a-yi) undecanamnide (68) and its l1cx-chlorocydopropyl and 17afluorocyclopropyl derivative (69) a. ,OMgBr L

CH

2 12 Zn/Ag

(CH

21

QCONC

4

H

9

CH

3

CX

3

CO

2 Na -l cX 2 (CI-I2) 1

CONC

4 Hg X=F, Cl 149 EXAMPLE 17 N-n-butyl, N-me thyI-(17'a-cy clop ropyl-3,1 7frihyd roxy 11 p-meLhoxy estra 1',3',5'(10')-trien-7a-y1) undecanamide (70 X 1-H) and its 17a-chlorocylopropyl and 17a-fluorocydopropyl derivative (70, X Cl) Same example as Example 16 with compound 49 as starting material.

o*.

C

6

HSCO'

)NQ

4 -i,70 '(CH 2 10

CONC

4 Hg CP.3 X F.CI C, -150 EXAMPLE 18 Scheme 18 l1ci-cyanovinyl derivatives N-n-buty, N-methyl-] l-(l7'a-cyclopropyl-3',17V-dihydroxy 11 W-methoxy estra S,Y',5'(10O)-trien-7ci-yl) undecanamide (73)

CH

3

I

C0 3 Na 2 a.

a.

a a a a.

(CH

2 10 C0NCI-I

OH

3

OH

71 1(HJ 0 0CH ~1

OH

3 1) HCE-CSi(CH 3 3 n-BuUj 2) KOH

B

(CHJ

10

CQNC

4

HN

B 1) MeUi,

CO

2 2) NH 3 3) NaBH 4 -pyr 4) H 2 Undlar cat- 151 EXAM1LE 19 Scheme 19 Compounds with aliphatic side chain in l1ca-position N-n-butyl, N-methyl-Il -(3',17prdihydroxy lTci-ethynyl estra I 0)-trien- 1 5'a-yl) undecanam-ide ("EM 108") 9 .9 9* *9 0 0 HO-I Ci)on Co 3 Na 2

H-

3 CO 74 C(HH01) 2 PhN(cH 3 2 Br 3 H3COCHC0CH 3 p sOH 0 Btvig(CH2)1 ITmp H3O06/ CUCI 0 3 77 H 3

CO

PTSA. MeOH 152 C00

(C,

1

H

t Co.

9 9 9O 99999.

1) C'CO 2

R

2) CI- 3

NHC

4

H-

9 1) NaBH 4 2) BBr 3

(CH

2 10

CONC

4

HN

EM 108 153 EXAMPLE Scheme 17a-thioethyl derivatives N-n-butyl, N-methyl-il -3',l7'-dihydroxy 17ci-thwoethyl estra

T

a-yI) undecanamide (82) and its ethyl disulfite derivative (83) .*0 0 :00..

0 0

H

2

S

(UYiJ 1

CONC

4

NI

82 (CH 2 10

C-ONC

4 Hj

CH

3

C

2

H

5

SOSC

2

H

*see 83 (CH 2 1

QCONC

4 Hg

CH

3 EXAMPLE 21 Scbteme 2-1 17a-thiopropyl derivatives N-n-butyl, N-methyl-I I-(Y,17'p-dihydroxy 17a-thopropyl estra-1 trien-T-a-yl) undecananiide (84) and its ethyl disulfite derivative (86) EtO 2

O-N=N-CO

2 Et PPh 3

S

0* S 0@ PhCOOH, THF (CH0), 0 C 6

H

5 COi CON(CH3)C 4

HN

OCOC

6

H

(CH2) 10

CON(CH

3

)C

4

H

9 NaOH 4% MeOH, PhCH 3 t amb.

EM 187

(CNI)

1

CON(CH

3 )C1-{ 155- EXAMPLE 22 Scheme 22 I I p-ethyl derivatives N-n-butyl, N-methyl-Il -{3',7'p-dihydroxy l7cz-ethynyl-I11P-ethyl estra I trien-Tcz-yl) undecanarnide (97) '9 9.

9 *9*9 *9 9 9 9999 9* 9 9* 999999

HO~

1) CH 3 1, N2CO 2) N HAlj 3) QAc 3 87 ."(CHNJ 1 OAc 1) C 5

H

6 N-Br 3 2) CId. DMF 3) (CH 3 2

C(CH

2

OH)

2

PTSA

OAC

0* QAc C 8 l)C H MgBr 2) Ac 2 O. pyr

H

5 OAc

HC

0 0 OH "/(CH 2 11 OAC 0 11 AcOBr. AcI,0 2)OH- 2 11 QAc

HSC

2

C

6

H

5

COCI

I"'(CK

2 11 0H

S

S

S

S. 55

S

4 5*SS

S.

5 5

S

S

S. S S *5

HSC

2 Jiones 1) CICO 2 R, n-BU3N 2) CH 3

NHC

4

H

9 I=OH C 6

HCO(J

1NaOH 2) NaBH 4 ONC,-L HO 1) HC-=CSi(CH 3 3 n-BuLi 2) KOH 157- EXANPLE 23 Scheme 23 14,15 epoxide derivatives N:n-butyl, N-me thy1- 1143', 1 7-dilbenzoyI-1 4',15'-epoxy-es tra 1',3',5'(I0')trien- 7a-yl) undecanarnide ("EM 180") and ("EM 181") 0

OH

1) PTSA PhiCH 3 H MA4

H

2) NaBH 4

Q

HO

HO

HO EM 112 (CH 2 10 C0N(CNC 4 Hg 9 (CH- 2 10

CON(CH

3

)C

4

HN

S.

S. S

S

S

S

S. *S 5

S

4

S

SS

OS S 5

S

S

S

5* 4.

Bc

C

5

H

5

N

0 2

CC

6

H

H

CAHC0 2 99 (CH 2 ),Q1CON(CH 3

)CAH

MCP8A CH 2

C$

r~zVl

'(CH

2 10

CON(CH

3

)C

4

H

9

C

6 HSC0 2

C

6

H

5 00j

'(CM

2 10 C01'(CH3)C 4

H

9 EM 180 EM 181 -158- EXAM{PLE 24 Scheme 24 N-n-butyl, N-methyl-I 1-(3',17'a-dihydroxy estra-l',3',5'(10') trien-TQ-yl) undecanamide ("EM 187") EtQ 2

O-N=N-CO

2 Et PPh 3 PhCOOH-, THF I mb 1 9 9 a.

9 2CC)C 6

HS

C

6

H

5

CO/-

NaOH 4% MeOH, PhCH 3 t. amb.

(CH

2 10 EM 187

CON(CH

3

)C

4

H

9 159 EXAMPLE Scheme N -n-butyl, N-methyl-ll (6*-hydroxy-2'-(4"-hydroxyphenyl)-3Y-ethyl-indol-N,-y1) undecanamide (104) The starting material 101 has been synthesized as described by Von Angered et al., J. Med. (Them. 27:.1439-1447, 1984.

NaH

-OCH

3

(CNIO

09- 101 102 2) CIOO -B 3) NH(CH 3 )0 4

H

9 BBr 3 (CH)j~OCON(CH 3

)C

4 N

H

I OH

(CH

2 10 C0N(CH 3

)C

4 Hg1 160 EXAMPLE 26 (Scheme 26) N-n-butyl, N-methyl-I I-(6'-hydroxy-2-(4"-hyciroxyphenyl)-(I ',2'-dehydronaphtalen-3'-yl) undecanamide (110) C0 2 Et 0NaH /(EtO)ffl

N.

6-Aodioxy-2-Tetralone R.1r (I eq) 105 R-CPF3CHCr1- 7 2) 8H- OH ~0 3)1- 1) BBr 3 2) aCOC0i-Bu.

NH(CHiJC 4 3) tNaOH 0

CON(CH

3 )CHg1 161 EXAMPLE 27 Scheme 27 N-n-butyl, N-methyl-i l-(4,4'-(1,2-diethyl-1,2-ethanydyl) bis-phenol-3-yl) undecanamide (115) TBD~ oq)

DMAP

TE3[MSOO 0 heeto QClONE

K

2 00 CH3GCHl 2 .l.OCONEt2 1) BW (I THF.

2)B(H~o TB DMSO 0 Z1)

H-

12) U.CH

TBDMSO

0

OH

'(CH

2 1

QCON(CH

3

)C

4 Hg 1) CICO iBu NH( ~i)nBu 0 2) NaOH HOo (CH 2 0 C00HHC HO1 162 Other sex steroid activity inhibitors in accordance with the invention may be synthesized by methods known in the art, by methods analogous to those set forth herein and modifying the syntheses set forth herein in a manner known in the art.

a** **ee 163- Without intending to be bound by theory, it is believed that diphenyl ethylene and diphenyl ethenylene nuclei discussed herein contribute an enhanced affinity for the estrogen receptor. The ethenylene version with the optional double bond present is preferred, as is a dosed third ring which includes, in its sides the ethenyl double bond and one side of one of the phenyl groups. The dosed third ring is exemplified by the ring, for example, of formulas XX and XXI below: *9 a

'R'(B-R

2 )xLG

XXI

Preferred side chains, which are believed to help provide antagonistic or inhibitory characteristics, include the RI[B-R 2 -1 L-G and A'[Y-A"]uXR 2 1 side -164 chains discussed and defined above. Preferred additional substituents to the nucleus are those whose presence facilitates synthesis or enhances stability of the compound, or its metabolic half-life without significantly retarding affinity for the receptor. Smaller substituents such as C 1

-C

2 alkyl or alkylene groups or halogens may be used.

The R 6 substituent of formula XX (the G 3 substituent of formula XXI) is preferably either hydrogen or a Ci-C 3 hydrocarbon such as methyl, ethyl or -*"propyl. Generally, this substitution location the atom receiving

R

6 of formula XX or G 3 of formula XXI) is the atom which is both one atom away from the A-ring and one of the two atoms which receives the optional double bond when it is present. In contrast, the preferred side chain RI(B- *o~e o

R

2 )LG of formula XX and R 1 00

LG

1

G

2 of formula XXI) is preferably substituted, :as shown in formulas XX and XXI, at the atom which is both one atom away from the optional pi bond and one atom away from Z. It is believed that a 44.. lower hydrocarbon substitution at the R 6 position of formula XX (the corresponding

G

3 of formula XXI or the R 6 position of formula I) substantially enhances the effectiveness of an inhibitor.

While both the A and D rings may be unsubstituted, certain embodiments include hydroxy substitution on one or both of the A and D rings (especially at the 3 and/or 12 positions of formula XXI or the 3 and/or 10 positions of formula XX). Other preferred substituents to the A and D rings include 165would be converted to hydroxy in vivo after the pharmaceutical is administered to a patient. Such substituents; include, for example, methoxy, ethoxy or esters.

-166- Set forth below are several preferred compounds for use in the pharmaceutical composition of the invention corresponding to the structure: 11

S

*0

XXI

EM Z R100 L G1 G2 G3 AorDring hydroxylated (at 3 or 2) EM 738 CH 2 (CH2) 5 CONC 8 CH 3

A+D

EM 681 CH 2

(CH

2 6 CONC 5

E

1 O

CH-

3

A+D

EM736 CH2 (CH2)7 -CON< CH 3 CH C 3

A+D

EM698 CH 2 (CH2) 8 -CON< CH 3

CH

9

CH

3

A+D

EM 819 CR 2 (CH2) 9 -CON< CH 3

C

4

H

2

F

7

C

2

H

5

A+D

EM 690 CR 2 (CH2) 10 -CON< CR 3

C

4 11 CH 3

A+D

EM661 CH 2 (Cl 2 5 -SO- C 4

H

2

F

7 C 2 H5 A+D EM 663 CH 2

(CR

2 6 -SO- CQH 9 CH 3

A+D

EM 654 CH 2 (CH2)7 -SO- C 5

HI

1 CH 3

A+D

EM 732

CH

2 (CH 2 9 C5H6F5

CH

3 I A I -SO- IA-iD J I I -167- EM 656 1 CH 2 (CH2 )lo I 4 I C2li5 A+D 0 *505 0* 0* 5* EM 360 OH 2 CH 4 O(CH2)2 OH 3

OH

3

CH

3 D only EM 431 OH 2

CH

2 C"HO(O1-2)2 NC- 5

H

10 C83 D only EM 363 OH 2

CH

2 C4X0CH 2 2

NC

4

H

8 0285 D only EM 471 CH 2 (CH2)5 CH3jO3- EM CH 2 (082)6 NC 5

H

1 O CH 3 A +D EM 465 CH 2 (CH2) NC2 4

H

8

C

2

HJ

5 A D EM 777 OH 2 C4HA(CH2) 2 -Nc OH 3

CH

3

CH

3 D only EM 773 CH 2

C

6 4

O(H

2 2

C

2

H

5

C

2

H

5 s 02H-5 D only EM 765 CH2 C4-84CH2) 2 NCSHIO M83 D only EM 778 OH 2

CGH

4 0CCH 2 2 NC4H 8 0285S D only EM 734 082 C 6 Hi 4 O(OH) OH 3 083 OH 3 D only EM 699 OH 2 C"H((CH) C 2

H

5 CzH5 083 D only EM 735 OH 2 C4-H 4 CX(CH2)2 NCSH 10 083 D only EM 725 082 061-14(XOH2)2 NC 4 83 OH 3 D only EM 779 OH 2

C"H

4 (0H 2 2

C

4

H

2

F

7

CH

3 H D only EM 542 0 068i40(082)2 CH 3 OH-, 083 A+D EM 543 0 06404C(082)2 N0 4

H-

8

OH

3

A+D

EM 562 0 C"H 4 CH2) 2

NC

5

H

10 01-3 A *D EM 756 0 04-140(082)2 NC 5

H

10

C

2

H

5 A +D EM 623 10 6 1-LO(CH2)3 FC 2

H-

5

C

2

H

5 01-3 A +D I I EM 321

C

4 0(CH-2) 3 NC4H1 0 CH 3

A+D

9* I.

S

S

EM m7 0 C"O4(CH2) 3 NC 5 I

C

2

H

5 A +DI oM69 C4HVO(Qi 2 3 13 1

CH

3

CR

3

D

EM 423 0 C&11 4 (CH2) 2 NC 5

HI

0 C 2

H

5 D only EM 428 0 C6W 4 OCH2) 2 NC41JB C~H D only oM43 C 6

H

4 0(0i2) 2 ICH3R' H3 Dol EM 472 A (H)3 CHO

CH-

3 D orly EM 492 0 C"HA(CH2)3

NC

4 Hs

C

2

H

5 D only EM 384 0 C",HO(CH-2)3

NCSH

10 H Neither EM 386 0 C6F 4 0(CH2I) 2 CH3 CH- 3

CH

3 Neither EM 382 0 C"HO(CH 2 3 CH 3 CH3 C 2

H

5 Neither EM 345 0 (CH2)7 -CON< CR 3

C

4

H

9 H A.D EM 4.53 0 (CH2)8 -CON< CH 3

C

4

H

2

F

7 013 A *D EM 358 0 (CH2)9 -CON< CR 3 C4H 9 H A*D EM 467 0 (012)10 -CON< CH3 C 4 1- 9

C

2

H

5 A +DU EM 532 0 (CH2)io CONC4-1 8

C

2

H

5 A +D EM 631 0 (CH2)5 NC4F%

CH

3

A+D

EM 721 0 (CH2)6

NC

5

HI

0

CR

3

A+D

EM 612 0 (CH2)7'

C

2

H

5

C

2

H

5

C

2

H

5 A +D EMS511 0 (CH2) 9 -SO- C 5

H

6

F

5 -H A +D EM 513 0 (Q-I2)io -So- C 4 1HIO

CH

3 A D

*S

S

S

*S S

S*

S

-16h9-

S.

0 0000 *0 S S

S

I S 0@@0 04 00 a o 0 a 0*O* a 0 0 *0O@ *4 0 4.

0*

S

4 EM 512 0 C 3 H12

NC

4

H

8 H- A+D EM 555 0 C 3

H

2 NC 5

H

10

CM

3

A+D

EM 560 S C 6

H

4 0CH2) 2

NC

5

H

10 H D only EM 635 1S C 64 W(CH2 2

NC

4

HI

8 CR D ony EM 547 IS CJ40(GH2) N C 2

H

5

C

2

H

5 H D ony EM 541 S (Cfi2)5 -CON< CH3 C 4

H

9 H A+D EM 634 S (CH2) -CON< CH 3

.C

4

H

9 G1 3

A+D

EM 563 S (CHA) -CON< CH 3

C

4

H

9 H A+D IEM 762 1S (CH2) -CON< 013

C

4

H

9 H

A+D

EM 941 S (CR 2 10 CONC 4

H

8 g CH 3 A +D EM 821 NCH 3

C

6 H4(CH2)

NC

5 1 0 H A+D EM 753 NCH 3

C&

4 0(CHRh IC 2

H

5 I C2H 5

CR

3

A+D

EM 637 Nd-I 3

CJH

4 0CH 2 3

NC

4

H

8

CR

3 A +D EM 343 0 C" 4 (XCI-I2 2

NC

5

H

10 Cl- 3 A i-D 170 Non-limiting examples of synthesis of representative inhibitors are set forth below.

EXAMPLE 28 scheme 28 CH30JOID-1--o ,oe FCCOCH3

CH

3 0J 2

COP

2

CH

3 3 CH3 0 4 (CHi 2 ),0TBDMS R, CO 2

CH

CH-

3 0 CQ2CH 3 "611

CH

3 0 (CH12),,TBDMS 0CH 3 CH3 0 (CH 2

),,OH

CH

3 0 12)1 nICONRaRb -171 P-methoxytetralone 1 (0.254 g ,1.4 mmol in 5 ml of THF) was added to a refluxed mixture of (MeO) 2 CO (2.8 mmol) and sodium hydride (38 mg) in THF ml). The mixture was heated at reflux during the night, cooled, water ml) was added and neutralized with 5% HCL and extracted with ether. The ether was washed with a cooled satured sodium bicarbonate and water. The organic phase was dried with anhydrous MgS04 and the solvent was removed under reduced pressure. The residue in THF (5 ml) was added to a mixture of sodium hydride (21 mg, .85 mmol) and RcI (CH3I ,8.5 mmol) in THF (15 ml) and HMPA (1.7 mmol). The mixture was stirred during the night, water (25 ml) was added and neutralized with 5% HCL and extracted with ether. The ether was washed with a cooled satured sodium bicarbonate and water. The organic phase was dried with anhydrous MgSO4 and the solvent was removed under reduced pressure and the compound 2 (Rc=CH 3 was obtained in the yield of .%.The action of MeOCOCN (0.1 ml, 10 min at -780C) on the compound 2 (Rc=CH 3 (200 mg in 5 ml THF) in presence of LDA (lithium diisopropyl amine prepared from 0.13 ml diisopropyl amine and 1.6 M butyl lithium 0.6 ml) at -25°C, 30 min) and HMPA (0.15 ml) yield in 83% the compound 3 (Rc=CH 3 To this compound (290 mg) in THF (20 ml) was added 35% potassium hydride (119 mg, 0.72 mmol 18-crown-6 (24 mg, 0.06 mmol) and the mixture stirred 8 h at 25 0 C and then I(CH 2 )80TBDMS (657 mg, 1.9 mmol) was added and the mixture heated at reflux 6h.The extraction with ethyl acetate yield the compound 4 Rc=CH 3 (98%).The decarboalkoxylation with lithium bromide in pyridine yield the compound 5 Rc=CH3 which was treated by -172the Grignard's reagent of 3-bromo-anisole followed by acidic treatment.The resulting compound 6 Rc=CH 3 was converted into amide 7 Ra=CH 3 Rb =C 4

H

9 Rc=CH 3 Thus a cooled solution compound 6 Rc=CH 3 (700 mg) in acetone (17 ml) was added Jones' reagent (8N-chromic acd solution, 0.77 ml).

After 30 minutes, isopropanol (5 ml) was added and the mixture was poured in water and extracted three times with ethyl acetate. The organic layer was washed twice with brine, dried over magnesium sulfate and evaporated to dryness. The crude acd was used in the next step without purification. To its solution in anhydrous methylene chloride (4 ml) at -10°C was added, under stirring, triisobutylamine (470 il, 1.96 mmol) and isobutylchloroformate (280 il, 2.1 mmol). After 40 minutes, N-methylbutylamine (1.5 ml) was added and the mixture was stirred at room temperature during 1 hour. Methylene chloride (50 ml) was added. The organic solution was washed with IN HC1, saturated sodium bicarbonate solution and water dried on magnesium sulfate and evaporated to dryness. The residue was purified by "Flash chromatography" on silica gel (Kieselgel 60, Merck, under 0.063mm, 50 Elution with a mixture of hexane-ethyl acetate gave the amide 7 Ra=CH 3 Rb =C 4 H9, Rc-CH 3 (63%) The removal of the protection into compound 8 (EM 736, n=7, Ra=CH 3 Rb=C4H 9

R=-CH

3 is performed with pyridine-HCI.

-173- TABLE I HO (CH 2 )nCONRaRb EMn Ra Rb EM 738 5 -4-8 EM 681 6 Clo EM 736 7 OH 3

C

4

H

9 EM 698 8 CIH 3

C

4

H

9 .EM 819 9 CH-3 C4-1 2

F

7 EM 690 .10 CH 3

C

4

H

9 0.0 174 EXAMPLE 29 Scheme 29 V .0 0 C113

'(CH

2 ),,OTs 6 9 momol 1 TBDMS: t-butyl dirnethyLsilyl MOM: Methyloxymethyl 175 The compound 6 Rc =CH 3 is deprotected with pyridine-HCI and the phenol function is selectively protected as MOM derivative by treatment with

CH

3

OCH

2 CI in pyridine. The esterification of the alcohol with MTCI in pyridine gives the compound 9 Rc CH 3 which is transformed in sulfide (n--9,Ra=CH 6

F!

5 Rc =CH 3 with sodium hydride and PRa, SI-I. The oxidation with mCPBA and the acidic hydrolysis gives the sulfoxide 11 (EM 732, n--9, Ra-C51H 6 F& Rc=CH 3 TABLE 2 HO

(CH

2 )nSORa a

EM

EM 661 EM 663 EM 654 ,EM 732 EM 656 5 6 7 9 110 EM 661 5 C 4

H

2

F

7 CH EM 663 6 C 4

H

9

CH

EM 654 7 C 5

H

11

CH

~EM 732 9 C 5

H

6

F

5

>C

3 Rc CH3 CH3 CH3 7 C2H5 EM 656 ilO

C

4

H

9

C

2

H

176- EXAMPLE Scheme ocx 0 b-tebralone R, CO, CH., S S a a S. S S S *Sa*S.

-O(C4'2J 2 NR.Rb

-O(CH

2 2 N RaRb -177- P-tetralone is alkylated with (MeO)2CO and sodium hydride in THF at reflux followed the reaction of sodium hydride and RI (CH 3 1) in THF and HMPA. The action of MeOCOCN in presence of LDA yield the compound 12 (Rc=CH 3 which is alkylated into compound 13 (Rc=CH 3 by IC61H40THP in presence of sodium hydride in THF and HMPA.The decarboalkoxylation with lithium bromide in pyridine followed by the modification of the phenol function (acidic hydrolysis and reaction with Cl (CH 2 2

NC

5

H

1 0 in presence of K 2 C0 3 gives the ketone 14 (RaRb=CSHO) which is transformed into compound 15 by reaction with the Grignard's reagent of Br CH 4 0TMS in ether. The acidic hydrolysis gives the compound 16 (EM 431, RaRb=CSHlo).

TABLE 3 EM Ra Rb

RC

EM 360

CH

3 CH 3 CH 3 EM 431

-C

5 Hjo-

CH

3 EM 363

-C

4

H

8 C 2

H

178 EXAMPLE 31 Scheme 31 MOO(Cl-I,)OTs MOMO 17 (CH 2 )nNRaRb HO (CH 2 )nNRaRb 18 ego* 179 The compound 17 R2Rb=-C5HjG-, Rc--CH 3 is obtained by the reaction of HNRaRbj and sodium hydride on the compound 9. The acidic hydrolysis gives the compound 18 (EM 473, RaRb-C5HO, Rc--CH 3 TABLE 4

~OH

S.

S

S

555* FEM n Ra Rb R EM 471 5 OH 3 0F1 3 0113 EM 473 6 -CsHio- 0113 EM 465 7 -04118- 02115 5*

S

S. S S 05 -180- EXAMPLE 32 Scheme 32 0 0 0000 00 S 0 000 0 0 0000 00 00 0 0 0 *050 0 00 00 0 000 0 0 **Oo 00 0 05 00 000500 0 HO ORd EQ-KBrMg Y,.a ,0,,,NRRb 181 The reaction of the o-tolualdehyde with the Grignard's reagent of BrCH 2 C4-1 4 0Rd (Rd TIf) followed by the oxidation of the resulting alcohol by PCC (pyridinum chlorochrornate) gave the compound 19. The reaction with OHCC6H 4

O(C-

2 2

NC

5 Hlo in presence of sodium hydride followed by bromination with N-bromosuccimidde on presence of light in CCI 4 gave the enone 20 (RaRb=-CSHjO)-). The product 21 (RaRb=-CSHIO-) is obtained by cyclisation with (L-Bu)3SnH and AIBN (2'-2'-azobisisobutyronitile) followed by reduction of the ketone with sodium borohydride and acidic removal of the protection and the alcohol function.The compound 22 (EM 735, RaRb=-C5H1IO-,

R&,--CH

3 was obtained by cyclisation with LDA followed by Grignarci's reaction with alkyl bromide and acidic removal of -the protection- 182 TABLE 0* 0S EXAMPLE 33 Scheme 33 0 0 HOHTH-PO Ot 23 24 0e 0 0 0 0*0* 0*00 0 0 0 0 *005 25

OR

THPO'

-184 EM 349 The compound 26 R=H.L Rc=H was reduced with sodium borohydride in ethanol and a mixture of resulting compound (300mg; .6mmol), 4-(2-chloroethyl)morpholine hydrochloride (267mg; 1.4mmol) cesium carbonate (9 7 8mg; 3.Ommol) and potassium iodide (100mg; 0.6mxnol) in NN-dimethylforrnamide was kept at 90*C and with stirring for 1h. Water was added and the resulting mixture was extracted several times with a mixture of ether and ethyl acetate Drying (MgSO 4 and removal of solvent gave a waxy material that was purified by chomatography on silica gel (hexanes: ethyl acetate; 3:7 a few drops of triethylamine) to yield the dehydrated derivativ of the compound 27 5 5(R=(CH 2 2

NC

4 Hi 8 0, Rc--H) (153mg; 41%).

S

6.

*see solution of above compound(153mg; 249pgmol) in a mixture of acetic add (60m1) and water (6m1) was kept at 100'C for 10min. The solvent was removed oleo under reduced pressure and the residue was purified by chromatography on silica gel (ethyl acetate: acetone; 3:1) to yield compound 28 (EM-349, 0* .R=(CH 2 2

NC

4

H

8 0, Rc=-H )(100mg; (5 NMR; 300MHz; solvent CD 3

O)D-

standard: TMS) 254 (4H; t- J 4.5Hz- cyclo,-N-CH 2 -CH2--O 2

-CH

2 2-73 t, J O-CH2-CH 2 3.66 (4H; t, J 4.5Hz; cydlo-N-CH 2

-CH

2 -CI-1 2

-CH

2 4.04 (2H; t; J 5.5Hz; O-CH 2 -Cl- 2 6.11 OIH; d; J 2.5Hz; CH Phenyl) 6.12 (1H; s; O-CH- Ph) 6.29 (1IH; dd; I 2.5Hz,8Hz; CH Phenyl) 6.69 (2H; d; J 8.5Hz; CH Phenyl) 6.78 (2H; d; J 8.5Hz; CH Phenyl) 6.94 (1 H; d; J 8Hz; CH- Phenyl) 6.95 (1 H; s; HC=C) 7.25 (2H; d; J 8.5Hz;- CH Phenyl) 731 (2H- d; J 8.5Hz;- CH- Phenyl).

185 TABLE 6

O(CH

2 )nNRaRb EM nl Ra Rb

R

99*EM 542 2

CH

3 MH 3 CR 3 EM 543 2

-CH-CH

3 EM 343 2

CQ

5 H-i

CR

3 EM 756 2 -CsHl(-

C

2

H

9:EM 623 3

C

2

H

5 C 2

H

5 CH 3 -EM 321 3

-C

4 HRi-

CH-

3 EM87M 3

-C

5 iH 10 C 2

H

EM 349 2 -C4H-80-

H

EM 691 3

CH

3

OH

3

H

-186- EXAMPLE 34 Same synthesis as example 33 in scheme 33, the compound 23 being replaced by the 2Z,4-dihydroxyphenylacetophenone TABLE 7

O(CH

2 )flNRaRb EM 432

H

EM 472OH Mn Ra Rb M432

C

2

H

l~ M482

-CH

3 8 Rc C2H5 CH3 CH3 CH3

CH

3

CH

3

-C

5 Hl 0 J -4 187- EXAMPLE Same synthesis than example 33 in scheme 33, the compound 23 b~eing replaced by the 2 '-hydroxypheny lace tophenone TABLE 8 C O(CH2)nNRaRb EM ni Ra Rb Rc EM 3&4 3 -051110-

CH

3 EM 386 2 0113 CH3 0113 EM 382 .3 CH3 13

C

2

H

a a. a a.

-188- EXAMPLE 36 EM 350 Synthesis described in scheme 33 Thus the compound 27 Rc=H) was reduced with sodium borohydride in ethanol and a mixture of resulting compound (300mg; 0.6mxnol), chioroacetic ::acid, piperidyl amide (242mg; L-5mmol) and cesium carbonate (978mg; ~.3.Ommol) in N,N-dimethylformamide (l0mi) was kept at 90*C and with o stirring for 1h. Water was added and the resulting mixture was extracted several times with ether. Drying (MgSO 4 and removal of solvent under reduced pressure gave a gummy residue that was purified by chomatography on silica gel (hexanes: ethyl acetate; 1:1) to yield compound 2 7

(R=CH

2

CONC

5 Hj 0 RC-=H) (127mg; 34%).

A solution of above compound (127mg; 203gmol) in a mixture of acetic acdd *(l0mi) and water (lml) was kept at 100*C for 10mmd. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 1:1) to yield compound 28 (EM 350,

R=CH

2

CONC

5

HI

0 (43mg; (8 NMP, 300MHz; solvent: CD 3

OD;

standard: TMS) 1.4-1.7 (6H; m; cyclo-N-CH 2

-CH

2

-CH

2

-CH-

2

-CH

2 3.40 (2H; t; J cyclo-N-CH 2 -CH2-CH2-CH2C-i2-) 3.49 (2H; t, J 5.5Hz; cyclo-N-CH 2

-CH

2

CH

2

-CH

2

-CH

2 4.67 (2K- s; 0-Cl-b-CO-N) 6.11 O1H; d; J 21-z; CH- Phenyl) 6.13 0 H; s; 0-Cl--Ph) 6.29 (1IH; dd; J 2Hz,8Hz; CH Phenyl) 6.69 (2H; d; j 8.5Hz; CH- Phenyl) -189 6.79 (2H; d; J 8.5Hz; CH Phenyl) 6.94 (1H; d; J 8Hz; CH Phenyl) 6.95 (1H; s; HC=C) 7.25 (2H; d; J 8.5Hz; CH Phenyl) 7.32 (2H; d; J 8.5Hz; CH Phenyl). Mass Spectroscopy: M+ 457 28 (CH 2 )nCONMeBu Rc=H) All these compounds were prepared by the following procedure. However, when n was superior to 1, potassium iodide was added to the reaction mixtures during the coupling reactions.

Typical procedure 28 (EM 357, n=l) Thus the compound 26 Rc=H) was reduced with sodium borohydridee in ethanol and a mixture of resulting compound (418mg; 0.84mmol), Nmethyl,N-butyl chloroacetamide (342mg; 2.09mmol) and cesium carbonate (136g; 4.18mmol) in N,N-dimethylformamide (20ml) was kept at 90 0 C and with stirring for 12h. Water was added and the resulting mixture was extracted several times with ether. Drying (MgSO 4 and removal of solvent under reduced pressure gave a gummy residue that was purified by chomatography on silica gel (hexanes: ethyl acetate; 1:1) to yield compound 2 7

(R=CH

2 CONMeBu, Rc=H) (276mg; 53%).

A solution of above compound 1 38mg; 220gmol) in a mixture of acetic acid and water (1ml) was kept at room temperature for 10min. The solvent was removed under reduced pressure and the residue was purified by -190chromatography on silica gel (ethyl acetate: hexanes;01:1) to yield compound 28 (EM 357, R= CH 2 CONMeBu. Rc=H) (38mg; (8 NMR; 300MHz; solvent:

CD

3 OD; standard: TMS) 0.85-1.0 (3H; m; N-CH 2

-CH

2 -CH2-CH 3 ).12-1.35 (2H; m;

N-CH

2

-CH

2

-CH-

2 1.4-1.65 m; N-CH 2

-CH

2

-CH

2

-CH

3 2-87 and 2-96 (3H; 2s; N-CH 3 3.25-3.4 (2H; m; N-CH 2

-CH

2 -C 2

-CHJ

3 4.66 and 4.68 2s; O-CH 2 CO-N) 6.12 (1H; d; J 2.51-z CH Phenyl) 6.13 s; O-CH-Ph) 6.29 dd; J CH Phenyl) 6.70 (211; d; J 8.5Hz; CH Phenyl) 6.78 and 6.79 2d; J CH Phenyl) 6.94 (1H; d; J 8Hz; CH Phenyl) 6.95 (1H; s; HCC) 7-25 d; J CH Phenyl) 732 (2HL- d; J 8.5Hz; CH Phenyl). Mass Spectroscopy- M+ 459 *i 191 TABLE 9 EM flRa Rb R EM 350 1-C 5

HI

0 H EM 357 I GH 3 (24H9 H EM 808 2 CH 3 (24H9 H EM 908 3 (21-3 (ZH 9

H

EM 901 4 01-3 (24H 9

H

192 EXAMI'LE 37 Scheme 34

'HP

0

OTHP

I &Ilnu I I f u X., THPO OH (CH2)n 30 0 NRaRb

S

S

S. *S S S *SS S

S

9*

S

0 29

THPI

31 0 NRaRb

,(CH

2 n+I NRaRb THPO' RaRb 193- EM 345 Synthesis described in scheme 34 A mixture of compound 24 (2.52g 6.12mmol), the aldehyde 29 Ra=Me, Rb=Bu) (1.00g; 4.08mmol) and piperidine (5001l) in benzene (170ml) was reflux for 48h by means of a dean-stark apparatus. The solvent was removed under reduced pressure and the resulting oil was purified by chromatography on silica gel (hexanes: ethyl acetate; 7:3) to yield the chalcone 30 Ra=Me, Rb=Bu) (620mg; 77% corrected yield) and the unreacted starting material 24 (2.00g).

To a solution of the chalcone 30 Ra=Me, Rb=Bu) (469mg; 0.73mmol) in ethanol (30ml) at room temperature. and with stirring was slowly added sodium borohydride (34mg; 0.89mol). The reaction mixture was stirred for a further 12h at room temperature. The solvent was removed under reduced pressure. The oily residue was taken in ethyl acetate and it was washed several times with saturated aqueous ammonium chloride solution. The organic extract was dried (MgSO 4 and solvent was removed under reduced pressure.

The residue was purified by chromatography on silica gel (hexanes: ethyl acetate; 4:1) to yield the chromene 31 R,=Me, Rb=Bu, Rc=H)(300mg; 66%).

A solution of compound 31 Ra=Me, Rb=Bu, Rc=H)(300mg; 482pmol) in a mixture of acetic acid (30ml) and water (3ml) was kept at 100 0 C for 30min. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel (ethyl acetate: hexanes; 1:4) to yield compound 32 194 (EM 345, n=7, R,=Me, Rb=Bu, RC=H) (104mg; (5 NMR; 30GM Hz; solvent-

CDQ

3 standard: TMS) 0.85-1.0 (3H; m; N-CH 2

-CH

2

-GH

2

-CH

3 ).1.15-1.9 (16H; m;

N-CH

2

-CH

2

-C-H

2 -CH3I and O-CH-(CH 2 6

-CH

2 -CO-N) 23-2.4 (2H; m; CH 2

-CO-N)

2-96 and 3.00 (31- 2s; N-Cl- 3 3-28 and 3.40 (2H; 2m; N-CH 2

-CH

2

-CH

2 -C-1b) 5.15 OIH; dd; J 2Hz,loHz; 0-C--Oi 2 6.44 (IH; dd; J 2.Hz,8Hz; CH Phenyl) 6-54 (1H; d; J 21U, CH Phenyl) 6.60 (1H; s; HC-C) 6.84 (2KH d; J 8.5Hz; CH Phenyl) 6.92 d; J 8H-17 CH- Phenyl) 7.31 d; J 8.51-174CH Phenyl). Mass Spectroscopy:. M+ 451.

195 TABLE 0 (CH 2 )nCONRaRb EM 45 EM 358 EM 467 EM 532 IM in Ra .Rb'-R

CH

3

CH

3

CH

3

C

4

H

9 0 4

H

2

F

7 C4119

C

4

H

9

H

C17 3

H

CH

3

C

2

H

-417q- M9 EXAMPLE 38 Typical procedure for compounds 34 (EM 371., n=10, Ra=Me, Rb=Bu, Rc--H) Synthesis described in scheme 34 To a solution of the an-ide 31 (n=10, Ra,=Me, Rb=Bu, Rc--H) (lO0mg; 0.15mmoI) in tetrahydrofuran (l0mi) under refiux and with §tirring was added a solution lithidum alumidnium hydride (1M in tetrahydrofuran; 0.42rn1; 0.42rnmol) The resulting mixture was refluxed for a fiurther 48h- 2N aqueous sodium hydroxide fee* solution was added to the reaction mixture and the aqueous layer was extracted *..*several times with ethyl acetate. The joined organic extracts were dried (MgSO 4 and solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (hexanes: acetone; 1:1) to yield the amidne 33 Ra=Me, Rb=Bu. Rc-H) (60mg; 62%).

A solution of above compound (60mg; 93pgmol) and pyridiniumn p- :toluenesulfonate (46mg; 185pimol) in methanol (l1rni) was refluxed for 12h.

a The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel (hexanes: acetone; 3:2) to yield the chromene 34 (EM 371, n=10, Ra=Me, Rb=Bu, Rc=H) (30mg; 67%) (8 NMR, 300MHz; solvent: CE}3OD; standard: TMS)O.93 (3H; t; J 7.5Hz; N-CH 2

-CH

2

-CH

2

-CH

3 ).1.15-l.85 (24H; m; N-CH 2

-CH

2

-CH

2

-CH

3 and O-CH-(CH 2 10

,-CH

2 2.24 (3H; s; N-Cl- 3 2.35-2.45 (4H; m; CH 2

-N-CH

2 5.19 0lH; dd; J 2.5H-z,10l-z; O-CH-CH 2 6.26 (11H; d; j CH Phenyl) 623 01-; cd;l J 2.5Hz,8.5Hz; CH Phenyl) 6.59 (1H; s; HC--C) 6.78 (2H;- 197 d; J 8.5Hz; CH Phenyl) 6.88 (IH; d; J 8Hz; Cli Phenyl) 7.30 J 8.5Hz; CH Phenyl).

C

BC

C 0

S

6C@*

C

S

e.g.

C. Ct C C

C

0@CC

C

COeC t C

CC..

C C

C

C

C. 0

CC

C CC

C

198 TABLE 11 HO1 0 (CH 2 )nNRaRb

I

EM

'Rb EM 631 5.

Em 721 6 EM 371 10 EM 612 7 a.' 3

C

2

H

5

CH

3 a.' 3

H

C2Hs C4-1 9 -199- EXAMPLE 39 Same synthesis as described in scheme 34 but the compound 29 is replaced by

OHC(CH

2 )nSORa and the reduction step was elimidnated.

TABLE 12 HO1 0 (CH- 2 )nSORa EM n Ra a EM 511 9

C

5

H-J

5 H EM-513 10 C41 1 0 CH- 3 S. S 200 EXLAMPLE Scheme 36

IH

-201 4'-hydroxy-2-(4-benzoyloxyphenyl) acetophenone 35 prepared from the commercial desoxyanisoin (250 g)(hydrolysis of the methoxyl group with pyridine-HCI at 220 OC followed by the treatment with 120 ml of benzoyl chloride and 20g of dimethylaminopyridine in 21 of methylene chloride during 24h) is bromined as usual manner with bromine in acetic acid at room temperature and the residue was without purification treated overnight with dihydropyran (150 ml) and p-toluenesulfonic acd (10 g) in 21 of methylene chloride at 0 OC.The bromo-compound 36 is isolated (170 g) after flash chromatography on silica gel in using ethyl acetate /hexane mixture containing 0.1% triethylamine. This compound is treated for 10 min at room temperature with sodium methylate in methanol followed with 2-mercaptobenzyl triphenyl phosphonium salt 37 (obtained from reduced commercial thiosalicylic acid and triphenyl phosphine). The resulting mixture was heated at reflux for 3 h with sodium methylate in methanol and the compound 38 (95 g) is isolated after ether extraction and flash chromatography on silica gel with ethyl acetate /hexane mixture containing 0.1% triethylamine. The benzyl protection is hydrolysed with sodium carbonate in methanol-water solution at room .:temperature and a mixture of resulting compound, N,N, diethyl chloroethylamine HCI and cesium carbonate in N,N-dimethylformamide ml) is kept at 90°C and with stirring for 12h, extracted several times with ether. Drying (MgSO 4 and removal of solvent under reduced pressure gives a gummy residue that is purified by chomatography on silica gel (hexanes: ethyl acetate; 1:1).The resulting compound in methanol (2 1) is heated at reflux with -202p-toluenesulfonic acid (5 g) for 3 h, Extraction with ethyl acetate and flash chromatography with silica gel (hexanes: ethyl acetate) gives the compo)und 39 (EM 547) (68 g) of wich structure was determined by spectroscopic means.

*.0 203 TABLE 13

H

i(CH 2 2 NRflRb, *.0 0*0* 0O 0* *o EMIR.p IEM 560 IEM 635 EM 547

C

2

H

5

.C

2

H

5

H

CH

3

H

204 0 0** 000 EXAMPLE 41 Same synthesis than the synthesis described in sheme 35 except the sta.rting coumpound 35 is HOC61- 4

CO(CH

2 )nCONRaRb TABLE 14 HO S (CH 2 )flCONRaRb EM nl Ra Rb R EM 541 5 C21 3

C

4

H

9

H

EM 634 7

CH

3

C

4

H

9

CH

3 E-M 563 8 C1-3 C 4

H

9

H

EM 762 9 CH 3

C

4

H

9

H

EM 941 10 -C4HS-

CH

3 '8*Nu('HO)O N~ OH 3* HO0 0 OO 0 wN ;v 0H II I j* N. 9.w 'HOOT'.0 tm 999 0 0 ~zt ef 3 ZqOx3 soz 206 The compound 40 obtained by acylation and rnethylation of the commercial manisidine is treated in a Friedel-Craft reaction with the 4 -methoxyphenylacetoyl chloride 41 and AlC1 3 A mixture of resulted compound 42, the aldehyde 43 (R=THP) and piperidine in benzene is refluxed for 48h by means of a dean-stark apparatus. The solvent is removed under reduced pressure and the resulting oil is purified by chromatography on silica gel. The condensed compound 44 is transformed into compound 45 by alkalin treatment (KOH in methanol) and the protecting group R is remplaced by (CH2) 2

NC

5 HlO0 by the method described in example 33. The resulting compound 45 is treated with methylmagne-sium go. iodide in THY followed by pyridine-HCl treatment to give compound 46 (EM O*P* 821, RaRb--CSjlO, Rc-01 3 s o* to 207 TABLE

I''

IRaRb EM nRa Rb R EM 821 2 -CsH 1 0

H

EM 753 2 C 2

H

5

C

2 Hs CH 3 EM 637 3 -C4HS. CH 3 S. S

S.

208 EXAMPLE 43 Scheme 37 COOH- CodI O

NH

2 MeOe a. MeO' a CH 2 0H- 0 ~HO CF M C2PMeO CH1 50 51 MeO' 209 The 4-methoxyphenylglycine 47 (prepared from commercial 4hydroxyphenyglycine) is treated with (COCI)2-Te resulting acid chloride 48 is used in a Friedel-Craft reaction on the 3-methoxybenzyl alcohol 49 in using aluminiurnchloride as catalyst. The coupled compound 50 is treated with methyl magnesium iodide in THF followed by the treatment with ptoluenesulfonyichioride in pyridine and CH 2

CI

2 and dlimethylaxninopyridine.The resulting compound 51 is heated and reduced with LiAlH 4 into the compound 52. The alkylation with NaH and Br(CH 2

)I

0 C0NC4H{ 8 in DMF gives the compound 53 (EM 877, RaRb=C 4

HS).

210 TABLE 16 0* a a a E-M nRa Rb

R

EM 865 6 C3C4H 9 CH 3 EM 876 9

CH

3 C 4

H

9 CH 3 EM 877 .10 -448-

H

-211- EXAMPLE 44 Same synthesis than the synthesis described in scheme 37. The compound 53 is reduced with LiA1H 4 in TI-I.

TABLE 17 The terms and descriptions used herein are preferred embodiments set forth by way of illustration only, and are not intended as limitations on the many variations which those of skill in the art will recognize to be possible in practicing the present invention as defined by the claims.

Claims (3)

1. A compound of the molecular structure: 0 wherein X 1 or 2; wherein R 5 and R 6 are independently selected from the group consisting of hydrogen, hydroxyl, and a moiety convertable to hydroxyl in vivo; and wherein R 5 and R 6 are not simultaneously hydrogen when X=l.

2. A pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and a S* compound according to claiml.

3. A method of treating breast cancer comprising administering to a subject an effective amount of a compound according to claim 1. Dated this 12th day of May 2000 25 ENDORECHERCHE, INC. By their Patent Attorney "GRIFFITH HACK ooo