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. Fo example, alternative or concurrent therapy to administration of antiestrogen could involve attempts to block the production of estrogens (e.g. b ovariectomy) such that less is available to activate receptor sites. However, prior art methods for blocking estrogen production insufficiently inhibi 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.

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 ma inhibit certain human breast tumor cells.

V

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

The combined use of the antiestrogen Tamoxifen and a luteinizin hormone-releasing hormone agonist, Buserelin, is also known for treatment o breast cancer. See, for instance, Klijn et al. J. Steroid Biochem. 420: no. 6B, 138 (1984). The objective remission of such cancers, however, remains unacceptabl low.

It has been found that certain 7α-substituted derivatives of estradiol, fo example a 7α-(CH2)ιoCONMeBu substitution possess antiestrogenic activit (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. Th substitution (CH2)9SOCsH6F5 has also been used on certain compound (Wakeling et al.. Cancer Res. 51: 3867-3873, 1991).

Certain -(CH2)ιoCONMeBu substituted compounds are also disclused in U Patent 4,732,912 (See e.g. example 5 and 16). See also EP Pat. No. 166509, EP Pa No. 124369, EP Pat. No. 160508, EP Pat. No. 163416, U.S. Pat. No. 4,760,061, U.S

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

Estradiol derivatives bearing a carboxyalkyl substituent at the 7α-positi maintained their affinity for the estrogen receptor when linked via th carboxy group to agarose or polyaαrylamide resin for affinity chromatograp purification of the estrogen receptor (Bucourt et al., J. Biol. Chem. 253: 82 1978).

Some steroid derivatives, such as 16-methylene estradiol and 16-methyle estrone, have been described as inhibitors of 17β-hydroxysteroid dehydrogen activity (Thomas et al., J. Biol. Chem. 258: 1500, 1983).

Certain nonsteroidal compounds which are stated to have antiandrogenic eff 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 f susceptible male animals including humans whose testicular hormon secretions are blocked by surgical or chemical means, e.g., by use of an LHR agonist, e.g., [D-Trp ⁶ , des-Gly-NH2 ¹⁰ ]LHRH ethylamide. The treatment includ administering an antiandrogen, e.g., flutamide in association with at least o inhibitor of sex steroid biosynthesis, e.g., aminoglutethimide and/ ketoconazole. See also PCT/U.S. 85/01454 (International Publication Numb

WO 86/01105) regarding combination therapy for treating hormonal-dependen cancers.

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

In U.S. Pat. No. 4,386,080 relates to new amide derivatives, and mor 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 llβ-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 17β-hydroxysteroid dehydrogenase, which is involved in the biosynthesis of testosterone, androst-5-ene-3β,17β-diol and estradiol, has been studied by some workers. Some affinity-label inhibitors for human placental estradiol

17β-dehydrogenase have been described (C.C. Chin and J.C. Warren, J. Bi Chem. 250: 7682-7686, 1975; Y.M. Bhatnagar et al., J. Biol. Chem, 253: 811-8 1978; C.C. Chin et al., J. Biol. Chem. 255: 3660-3664, 1980; J.L. Thomas and R. Strickler, J. Biol. Chem. 258: 1587-1590, 1983).

B. Tobias et al., J. Biol. Chem. 257: 2783-2786 (1982) and R.J. Auchus and D. Covey, Biochemistry 25: 7295-7300 (1986) disclose, respectively, the use 17β-propynyl-substituted progestins and propynyl-substitut 3-hydroxy-14,15-secoestra-l,3,5(10)-trien-17-one as inhibitors of the 17β-estradi dehydrogenase.

Thomas J.L. et al., J. Biol. Chem. 258: 11500 (1983) have described th 16-methylene estradiol and 16-methylene estrone are inhibitors 17β-hydroxysteroid dehydrogenase activity.

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

Von Angerer et al. discuss other antiestrogens in "l-(aminoalkyl)- phenylindoles as Novel Pure Estrogen Antagonists", J. Med. Chem. 1990; 3 2635-2640. In U.S. Patent 4,094,994, where it is said that the use of certai 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., Med.Chem. 33: 3216-3222 and 3222-3229 (1990) described the synthesis an

biological activities of 2,3-diaryl-2H-l-benzopyrans analogs as antiestrogen having the following molecular structure:

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

The European counterpart of priority applications 07/377,010 and 07/265,15 was published on May 9, 1990 as European Application No 0367576. Th 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 17 acyl 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 fo drugs. The Search Report emphasized Examples 2, 3, pages 22, 23 and th claims.

In D.E. Patent No 3242894 Al, Neef et al relates to 17α-substituted equilenin f inhibition of progesterone biosynthesis and control of the fertility.

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

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

Other Search Report References were previously discussed herein.

OBJECTS OF THE INVENTION

It is an object of the present invention to provide methods of inhibiting se steroid activity. Such methods may be useful in the treatment of se steroid-related diseases.

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

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

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

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

It is another object of the invention to provide inhibitors of sex steroid activit useful in the treatment of both estrogen- and androgen-related disease Androgen-related diseases include but are not limited to prostate cancer, acn

vulgaris, hirsutism, precocious puberty, benign prostatic hyperplasia, seborrhe androgenic alopecia and sexual deviants. Control of androgen activity may als be useful in male contraception.

SUMMARY OF THE INVENTION

In accordance with the invention, a pharmaceutical composition is provid comprising a therapeutically effective amount of the diphenylethyle derivatives specified herein. One embodiment of the diphenylethyl framewo is illustrated below:

where the dotted line represents an optional double bond of Z or configuration. Certain preferred substituents include, but are not limited to t following:

R _l , R5, Re, and R12 are preferably independently selected from the grou consisting of hydrogen, hydroxyl, halogen, lower alkyl, lower alkox alkylsulfonyl lower alkoxy, arylsulfonyl lower alkoxy, lower alkylsilyl, amin nitro, nitrile and nitroso.

R2, _/ , R ₉ and Rn are preferably independently selected from the grou consisting of hydrogen, hydroxyl, halogen, lower alkyl, lower alkox alkylsulfonyl lower alkoxy, arylsulfonyl lower alkoxy, lower alkylsilyl, amin nitrile, nitro, nitroso, azido, (C -C7) alkanoyl mercuryl, lower alkylamin dilower alkylamino, AXR ₂ 1, YT-AH -AΠJ _U -XR∑I, and Al[Y-A"] _u -XR ₂ ι wherein:

A is straight- or branched-chain (C1-C30) alkylene, (C2-C30) alkenylene, (C2-C30) 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. A ¹ and A ¹¹ 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 A ¹ and A ¹¹ together have a total of from 3 to 20 carbon atoms and Y is selected from the group consisting of -0-, -S-, -Se-, -SO-, -SO2-, -CO-, -NR22-, SiR22R22- -CR22OR22-/ -NR22CO-, -NR22CS-, -CONR22-, -COO- -COS-, -SCO-, -CSS-, -SCS-, -OCO- and phenylene (R22 being hydrogen or lower alkyl), wherein R2 ₁ is selected from the group consisting of hydrogen, straight- or branched-chain lower alkyl, lower alkenyl or lower alkynyl, (C3-C7) cycloalkyl, halogeno(lower)alkyl, carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower)alkyl, (C ₆ -C ₁₀ ) aryl, (Cβ-Cio) arylalkyl,

di(lower)alkylamino(lower)alkyl and fluoro-substituted analogs of the foregoing, and wherein X is selected from the group consisting of -CONR ₂ 3-,-CSNR ₂ 3-, -NR24CO-, -NR24CS-, -NR24CONR23-, -NR24-C(NR ₂ 5)-NR23, -SO2NR23-, -CO-, -CSS-, -SCS-, -O-, -NR23-, -(NOR23-, -(PO)R23-, -NR24COO-, -NR24SO2-, -S-, -SO- and -SO2- (R23 being selected from the group consisting of hydrogen, lower alkyl, a species which, together with R21, 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 R24 being hydrogen or lower alkyl, and R25 being hydrogen, nitrile or nitro). In certain preferred enbodiments, XR21 forms a tetrazole ring, CON CχH2x _/ (where x is an integer from 4-6).

R3 and Rio are preferably independently selected from the group consisting hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy, lower alkox carbonyloxy, carboxyl, ( -C20) alkanoyloxy, (C3-C20) alkenoyloxy, (C3-C2 alkynoyloxy, (C7-C1 ₁ ) aroyloxy and alkylsilyloxy.

Rό and R7 are preferably independently selected from the group consisting hydrogen, amino, lower alkylamino, dilower alkyl amino, nitro, nitril nitroso, halogen, lower alkyl, lower alkenyl, lower alkynyl, halogeno low alkyl, halogeno lower alkenyl, halogeno lower alkynyl, alkyl sulfonyl, ar

sulfonyl, a substituted 5 to 7 member heterocydic ring having at least o heteroatom (selected from oxygen, sulfur, silicon, selenium, nitroge -CH2)sW (wherein W is nitrile, hydroxyl, azido, nitroso, alkoxy, nit thionitrile, halogen, alkyl sulfonyl or aryl sulfonyl and s is an integer from 1 6), 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 R ι is hydrogen, hydroxyl, halogen, lower alkyl, lower alkenyl, lower alkynyl, nitrile, nitro, nitroso or Xg CH2)n 6 (Xό being selected from the group consisting of -O-, -S-, -Se-, -SO-, -SO2- and -CO-, and Y6 being selected from the group consisting of hydroxyl, amino, monoalkyl amino, dialkyl amino, 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),

AXR21, Y7-A ¹ -LY-A ¹¹ ] _u .-XR ₂ ι, and A^-[Y-A ⁿ ] _u -XR2l, wherein:

A is selected from the group consisting of straight- or branched-chain (Ci -C ₃₀ ) alkylene, (C ₂ -C ₃₀ ) alkenylene, (C2-C30) alkynylene and fluoro-substituted analogs of the foregoing, wherein u is an integer from 0 to 5, wherein Y7 is absent or is selected from the group consisting of carbonyl, carboxyl, -CH2S- and -CH2O-, wherein A- and A ¹¹ may be the same or different and may be 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 A ¹ and A ¹¹ together have a total of from 2 to 30 carbon atoms, wherein Y is selected from the group consisting of -O-, -S-, -Se-, -SO-, -SO2-, -CO-, -NR22-, -SiR22 R22-, -CR22OR22-; -NR22CO-, -NR22CS-, -CONR22-, -CSNR22- -COO-, -COS-, -SCO-, -CSS-, -SCS-, -OCO- and phenylene (R22 being hydrogen or lower alkyl), wherein R21 is selected from the group consisting of hydrogen, straight or branched chain lower alkyl, lower alkenyl, lower alkynyl, (C ₃ -C7) cycloalkyl, halogeno (lower) alkyl, carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower) alkyl, (C ₆ ~Cιo)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 -CONR23-, -CSNR23-, -NR24CO-, -NR24CS-, -NR24CONR23-, -NR ₂ 4-C(NR ₂ 5)-NR ₂ 3- _/ -SO2NR23-, -CO-, -CSS-, -SCS-, -O-, -NR ₂₃ -, -(NO)R23-, -(PO)R ₂ 3-, -NR24COO-, -NR24SO2-, -S-, -SO- and -S0 ₂ - (R23

being selected from the group consisting of hydrogen, lower alkyl and a spedes which, together with R2 ₁ , forms a saturated or unsaturated heterocydic 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, R24 being hydrogen or lower alkyl and R25 being hydrogen, nitrile or nitro). In certain preferred embodiments, XR21 forms a tetrazole ring.

Rβ and R7 may also be a species which, in combination with anoth substituent of general molecular formula I, forms a moiety selected from t group consisting of: -CH2-, -CHX-, -CX2- (X being halogen, carboxyl alkoxycarbonyl), -O, -S-, -Se-, >N-CN, >NR∑9 and >NCθ2R2 - (R29 bei hydroxy or lower alkyl), lower alkylene, -(CH2)rO(CH2)s-, -(CH2) _r S(CH2) -(CH ₂ )rSe(CH ₂ )s-, -(CH ₂ )rSO(CH ₂ )s-, -(CH ₂ )rSθ2(CH ₂ )s-, (CH ₂ )rCO(CH ₂ )s -(CH ₂ ) _r NR22(CH2)s-, -(CH ₂ )rSiR22R22(CH2)s- or -(CH ₂ )rCR22(CH2)s- (R22 bei hydrogen or lower alkyl, r and s being independent integers from 0 to 3), moiety of the formula:

wherein

A ¹ , Y, A ¹¹ , u, X and R21 are as defined above, wherein Z is absent or is select from the group consisting of lower alkylene, halogeno low alkylene,-(CH2) _n O-, -(CH _∑ JnS-, -(CH2) _n Se-, -(CH _∑ VSO-, -(CH2) _n Sθ2-, -(CH2) _n C -(CH2) _n NR22- -(CH2) _n SiR22R22- and - CH2) _n CR22θR22-, Rl2 is as defined abo n being an integer from 0 to 3, and R7 ₁ being selected from a group consisting hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy and lower alkylsilyl, moiety of the formula:

wherein N is nitrogen atom and A ¹ , Y, A ¹¹ , u, X and R21 are as defined above.

In preferred embodiments, moieties which are combinations of R groups fro general structure I, are combinations of R and R7, Re with R or R12, or R7 wi R5 or Rs.

The invention further provides an inhibitor of sex steroid activity having, part of its molecular structure, a substituted or unsubstituted estrogenic nude of general formula V:

wherein R ⁵ and R ⁶ are hydrogen lower alkyl, alkoxy carbonyl, (C1-C20) alkano (C3-C20) alkenoyl, (C3-C20) alkynoyl, (C7-C11) aroyl and alkylsiiyl,

wherein dotted lines are optional pi bonds. In some embodiments, the option pi bonds are not simultaneously present when aromatiάty would result fro 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 lowe alkyl substituent, wherein e is selected from the group consisting of carbo sulfur and nitrogen, q is absent or is a divalent methyl or ethyl moiety; sai inhibitor further having a side chain of the formula -RH-B-RZ-J _X L-G wherein i at least one of said side chains is substituted at a position selected from th group consisting of carbon 2, carbon 4, carbon 5, carbon 10, carbon 11, carbon 1 q and e atom wherein:

x is an integer from 0 to 6, wherein at least one of L and G is a polar moi distanced from the substitution point by at least three intervening atoms, a wherein:

R ¹ and R ² are independently either absent or selected from the group consisti of straight- or branched-chain alkylene, straight- or branched-diain alkynyle straight- or branched-chain alkenylene, phenylene, and fluoro-substitut analogs of the foregoing;

B is either absent or selected from the group consisting of -0-, -S-, -Se-, -S -SO ₂ -, -NR3-, -SiR ³ ₂ -, -CR30R3-, -NR^CO-, -NR3CS-, -CONR3-, -CSNR ³ -, -CO -COS-, -SCO-, -CSS-, -SCS-, -OCO- and phenylene (R ³ being hydrogen or lo alkyl);

L is either a moiety which together with G, forms a heterocydic ring having least one nitrogen atom or is selected from the group consisting of lower alk -CONR ⁴ -, -CSNR4-, -NR5CO-, -NR5CS-, -NR5CONR ⁴ -, -NR5C(NR6)-NR -S0 ₂ NR ⁴ -, -CSS-, -SCS-, -(NO)R ⁴ -, -(PO)R ⁴ -, -NR5COO-, -NR5S0 -, -0-, -NR -S-, -SO- and -SO2- (R ⁴ and R ⁵ being independently selected from the gro consisting of hydrogen and lower alkyl; and R ⁶ being selected from the gro consisting of hydrogen, nitrile and nitro); and

G is either a moiety which together with L forms a heterocydic ring having least one nitrogen atom or is selected from the group consisting of hydroge

lower alkyl, lower alkenyl, lower alkynyl, (C ₃ -C7) cydoalkyl, bromo(lower)alk chloro(lower) alkyl, fluoro(lower)alkyl, iodo(lower)alkyl, cyano(lower)alk carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower)alkyl, (C6-C1 ₀ ) aryl, (C Cn)arylalkyl, di(lower)alkylamino(lower)alkyl, fluoro-substituted analogs the foregoing.

This invention furtiier provides a pharmaceutical composition comprising pharmaceutically acceptable diluent or carrier and a therapeutically effecti amount of the foregoing sex steroid activity inhibitor.

The inhibitor is preferably hydroxy-substituted in at least the 3 or 12 position and is preferably substituted at the 7 position with a C1-C4 alkyl. Compounds formula V above may be used, preferably as part of pharmaceutic compositions induding acceptable diluents or carriers, to treat sex steroi dependent diseases by inhibiting sex steroid activity.

In another embodiment, the invention provides a pharmaceutical compositio comprising a pharmaceutically acceptable diluent or carrier and therapeutically effective amount of a sex steroid activity inhibitor having, a part of its molecular structure, an estrogenic nudeus of the formula:

wherein said sex steroid activity inhibitor includes, as another part of i molecular structure, a side chain of the formula R ¹ (B-R ² ) _X LG substituted onto ring carbon of said estrogenic nudeus to form:

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

R ¹ and R ² are independently either absent or selected from the group consisti of straight- or branched-chain alkylene, straight- or branched-chain alkynylen straight- or branched-chain alkenylene, phenylene, and fluoro-substitute analogs of the foregoing;

B is either absent or selected from the group consisting of -O-, -S-, -Se-, -S -SO2-, -NR30-, -SiR30 ₂ -, -CR30OR30-. -NR3CO, -NR30CS-, -CONR30-, -CSNR30 -COO, -COS-, -SCO, -CSS-, -SCS-, -OCO and phenylene (R30 being hydrogen lower alkyl);

L is either a moiety which together with G, forms a heterocydic ring havin least one nitrogen atom or is selected from the group consisting of lower al -CONR ⁴ -, -CSNR ⁴ -, -NR5CO-, -NR5CS-, -NR5CONR ⁴ -, -NR5C(NR60)N -SO2NR ⁴ -, -CSS-, -SCS-, -(NO)R -(PO)R -NR5COO, -NR5COO, -NR5S -O, -NR ⁴ -, -S-, -SO, and -SO2- (R ⁴ and R ⁵ being independently selected fr the group consisting of hydrogen and lower alkyl; and R ⁶⁰ being selected fr the group consisting of hydrogen, nitrile and nitro);

G is either a moiety which together with L forms a heterocydic ring having least one nitrogen atom or is selected from the group consisting of hydrog lower alkyl, lower alkenyl, lower alkynyl, (C3-C7)cydoalkyl, bromoGower)alk chloro(lower)alkyl, fluoro(lower)alkyl, iodo(lower)alkyl, cyano(lower)alk carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower)alkyl, (C6-Cιo)aryl, ( Cn)arylalkyl, di(lower)alkylamino(lower)alkyl, and fluoro-substituted analo of the foregoing;

wherein Z is selected from the group consisting of lower alkylene, haloge lower alkylene, -(CH ₂ ) _n O, -(CH2) _n S-, -(CH ₂ ) _n Se-, -(CH ₂ ) _n SO, -(CH ₂ )nS0 -(CH2) _n CO, -(CH2) _n NR22- -(CH2) _n SiR22R22- and -(CH ₂ ) _n CR22θR22 (wherein R 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 hydrogen, hydroxyl, halogen, lower alkyl, lower alkoxy, lower alko carbonyloxy, carboxyl, (C1-C20) alkanoyloxy, (C3-C20) alkenoyloxy; (C3-C2

alkynoyloxy, (C ₇ -C ₁₁ ) aroyloxy alkylsilyloxy; OR' ₃ , or OR' ₁₀ (whererin R' ₃ a R' _l O are hydrogen, alkyl, (C1-C20) alkanoyl, (C3-C20) alkenoyl, (C3-C20) alkyno or (C7-C ₁ 1) aroyl); and

wherein R ⁶ is selected from the group consisting of hydrogen, lower alk lower alkenyl and lower alkynyl.

when L and G are together a nitrogen hetero ring, such ring is preferab or -N χ-iH2χ-2θ (where x is an integer from 4-6).

R6 is preferably methyl, ethyl or propyl. Unsaturated analogs such as ethenyl ethynyl may also be used. In some embodiments, at least one of the A and rings is substituted with hydroxyl or a moiety converted in vivo to hydrox (e.g. hydrogen in positions 3 and 10, acetoxy, benzoyloxy, akanoylox alkenoyloxy and aroyloxy). For example, R ³ and/or R ¹⁰ may be hydroxyl. T ring-closing moiety, Z, is preferably -O-, -S-, -NH- or -CH2-, and in so embodiments is a bivalent moiety containing carbon, sulfur or nitroge Preferred side chains (e.g. R ¹ (B-R ² ) _X LG) are discussed in the detailed descriptio and examples herein.

In another embodiment, the estrogenic nucleus may indude a ring nitro onto which ϊs substituted the side chain. Thus, the invention provide pharmaceutical composition comprising a pharmaceutically acceptable car or diluent and a therapeutically effective amount of a compound of formula:

wherein the dotted line represents an optional double bond,

wherein R5 and ^ are independently hydrogen, hydroxy or a moiety which converted to hydroxy in vivo;

wherein R ¹⁰⁰ is bivalent moiety which distances L from the B-ring by 4- intervening atoms;

wherein L is a bivalent or trivalent polar moiety selected from the gro consisting of -CO, -SO, -CON<, -N< and -SON<

wherein G ^l is either absent or selected from the group consisting of hydroge a Ci to C ₅ hydrocarbon, a substituted or unsubstituted C ₅ to C ₇ cydoalkyl, bivalent moiety which joins G ² and L to form a 5- to 7-membered heterocyd ring, and halo-substituted derivatives of the foregoing; and

wherein G ² is either absent or selected from the group consisting of hydrogen, Ci to C5 hydrocarbon, a substituted or unsubstit ted bivalent moiety whi joins G ¹ and L to form a 5- to 7-membered heterocydic ring a halo-substituted derivatives of the foregoing.

In another embodiment, the invention provides a pharmaceutical compositi comprising a pharmaceutically acceptable carrier or diluent and therapeutically effective amount of an estrogen activity inhibitor of t following formula:

wherein the dotted line is an optional double bond;

wherein R5 and Re are independently hydrogen, hydroxyl or a moiety which converted to hydroxyl in mm;

wherein Z is a bivalent ring closing moiety.

wherein R ¹⁰⁰ is a bivalent moiety which distances L from the B-ring by 4- intervening atoms;

wherein L is a bivalent or trivalent polar moiety selected from the gro consisting of -CO, -SO, -CON<, -N< and -SON<.

wherein G ¹ is either absent or selected from the group consisting of hydrogen, Ci to C5 hydrocarbon, a saturated or unsaturated C5 to C7 cydoalkyl, a bivale moiety which joins G ² and L to form a 5- to 7-membered heterocydic ring, an halo-substituted derivatives of the foregoing;

wherein G ² is either absent or selected from the group consisting of hydrogen; Ci to C5 hydrocarbon, a substituted or unsubstituted C5 to C7 cydoalkyl, bivalent moiety whidi joins G ¹ and L to form a 5- to 7-membered heterocydi ring and halo-substituted derivatives of the foregoing.

In one embodiment, Z indudes a carbon, sulfur or nitrogen atom. In anothe

.«,. G ¹ embodiment, Z in -CH2-, -NH-, -S- or -O. In the side diain R™°L< _G2 , it i

preferred that at least one of G ¹ and G ² has at least two carbon atoms. G ¹ an G ² , in some embodiments, are independently absent or are selected from th group consisting of hydrogen, Ci to C5 hydrocarbon, a substituted o unsubstituted C5 to C7 cydoalkyl and halo-substituted derivatives of th 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 intervenin atoms. Unsaturation in R ¹⁰⁰ may include, for example, phenylene o alkynylene. Preferred moieties for R ¹⁰⁰ indude but are not limited to R ¹ (B-R ² ) from the side chain R ¹ (B-R ² ) _X LG discussed above or A'-(Y-A") _U from the sid

chain A'-(Y-A")uXR ₂ 1 discussed above. Preferred side chains include but a not limited to -(CH2)ιoCONCH ₃ C ₄ H9, -(CH ₂ ) ₆ NC ₅ Hι ₀ an -0-O(CH2)2NCsHio. When G ¹ , G ² and L combine to form a nitrogen-containin heterocydic moiety, such moiety is preferably, but is not limited to, or (where x is an integer from 5-7).

As used herein, the term "sex steroid activity inhibitor" includes an compound which suppresses the activity of sex steroids by any mechanis induding, for example, inhibition of sex steroid synthesis or antagonisti blocking of sex steroid receptors. "Androgen activity inhibitors" and "estroge activity inhibitors" are sex steroid inhibitors capable of inhibiting the activity androgens and estrogens, respectively. For example, estrogen activity inhibito indude, but are not limited to antiestrogens which block estrogen receptor thereby making them unavailable to estrogen compounds which coul otherwise activate those receptors. Sex steroid activity inhibitors also indud compounds which inhibit the formation of compounds capable of activatin sex steroid receptors such as inhibitors of the production of natural sex steroid (e.q. 17β-estradiol) or inhibitors of production of precursors of natural se steroids. One mechanism by which these sex steroid production inhibitors ma operate is by blocking enzymes which catalyze production of natural se steroids or their precursors (e.g. inhibitors of enzymes such as aromatas 17β-hydroxysteroid dehydrogenase, 3β-hydroxysteroid dehydrogenase and th like).

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

The following conventions apply to structural formulae set forth herei Unless specifically designated to the contrary, substituents may have either o β stereochemistry or, where valence permits may represent one substituent in- position and another in β position. Presence of optional pi bonds a independent of each other. All structures indude salts thereof. Atoms of a estrogenic nucleus for which no substituent is shown or described m optionally be substituted or unsubstituted so long as such substitution does n prevent the nudeus from functioning as an "estrogenic nudeus" as define herein. Those atoms having a defined substituent may optionally be furth substituted by other substituents where their valence permits such furth substitution. As used herein, the term "lower", when describing a chemic moiety means a moiety having 8 or fewer atoms. For instance, a "lower alky means a _. to Cg alkyl. Any moiety of more than two atoms may be straight- branched-chain unless otherwise spedfied.

BRIEF DESCRIPTION OF DRAWINGS

Figure 1 illustrates a competition binding assay of the affinity of estradi diethylstilbestrol, ICI 164384 (Wakeling, A.E .and Bowler, J., 1987; J. Endocrino 112: R7-R110) and EM-142 (an antiestrogen having a nonsteroidal nudeus an synthesized in example 1, herein) for the rat uterine cytosol receptor (Asselin al., 1978; J. Steroid Biochem. 9: 1079-1082).

Figure 2 illustrates the effert of the indicated doses of EM-142 injected twi daily (b.i.d.) on uterine weight (mg) in adult female ovariectomized Balb/ mice treated for 4.5 days in the presence or absence of simultaneous treatme with 17β-estradiol (0.01 μg, b.i.d.).

Figure 3 illustrates the effect of the administration of EM 139 on uterine weig (mg) in adult ovariectomized mice. The compound was administered twi daily (b.i.d.) at the indicated doses for 4.5 days in the presence or absence of 17 estradiol (0.01 μg, b.i.d.) via the subcutaneous route.

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

Figure 5 illustrates the comparative inhibitory activity of increasin concentration of EM 343 (•-•) and EM 312 (o-o) on the growth of human brea cancer ZR-75-1 cells stimulated by 17β-estradiol. The respective IC50 values ar calculated at 2.55 x 10" ¹⁰ M for EM 343 and 8.43xlO- ¹⁰ M for EM 312, th indicating a 3-fold higher activity for EM 343.

DEΓAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

In certain preferred embodiments of the invention, the R3 and Rio substituents on the nudeus of structure I supra are hydroxyl, (C1-C20) alkanoyloxy (C3-C20) alkenoyloxy, (C3-C20) alkynoyloxy, (C7-C10) aroyloxy and/or the R7 substituent is It is also preferred that the R7 substituent have between 7 and 20 carbon atoms. It is also preferred that of structure I, supra be lower alkyl, ethyl, fluoroethyl, or (CH _∑ hW, 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:

wherein A ¹ , A ¹¹ , Y, u, X and R ₂ ι are defined as previously for R^ and R7 in the formula I, wherein the double bond is in trans configuration,

wherein R ₃ and R' ₁₀ are hydrogen, alkyl, ( -C ₂ 0) alkanoyl (C ₃ -C20) alkenoyl, (C3-C ₂ 0) alkynoyl or (C7-Cn)aroyl,

wherein ^ is preferably selected from the group consisting of hydrogen, nitro, 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 heterocydic 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 6), or has the formula:

wherein:

F is present 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 wherein X$ is selected from the group consisting -O, -S-, -Se-, -SO, -SO2- and -CO and Y ₆ is selected from the group consisting hydroxyl, amino, monoalkyl

amino, dϊalkyl amino, dimethyl N-oxide, N-aziridyl, guanidino, N-pyrrolidino, N-piperidino, N-methylpiperazϊno, N-morpholino and alkoxy, and n is an integer from 1 to 6 preferable 3.

R9 and Rn 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, AXR21 and A^Y-A^Ju-X^i, wherein A, A ¹ , A ¹¹ , Y, X, R21 and-u are as defined previously from 6 and R7.

When administered systemically, pharmaceuticals of the inventions may be used in the treatment of breast cancer, uterine cancer, ovarian cancer, endometriosis, uterine fibroma, precodous 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 (e.g. sex steroid activity inhibitor), per day per 50 kg of body wdght, most preferably from about 10 mg to about 100 mg per day per 50 kg of body wdght

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

included 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 indude 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 indudes 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 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 suffident 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.

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 μg/ml and 10 μg/ml.

In certain alternative embodiments, the pharmaceutical composition of the invention may be foπnulated 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.

Other alternative preferred embodiments indude pharmaceutical compositions comprising therapeutically effective amounts of compounds of the formula:

wherein the dotted line represents an optional pi bond, wherein R'3, R'IO, A ¹ , Y, A ¹¹ , X,-R2i and u are defined as previously in formula VI espedally A*-[Y-A"]u-XR21 s -COp-C6H4-0(CH2)nCONR ₂ ιR22;

wherein R ₂₁ and R ₂₂ are defined as previously for R or R7 in formula I and n is an integer from 1 to 15; or:

wherein the dotted line represents optional double bond, espedally in trans configuration, wherein R3, R'ιo, Rό are defined as previously, wherein R21 is selected from the group consisting of hydrogen, straight- or branched-chain lower alkyl, lower alkenyl or lower alkynyl, (C3-C7) cydoalkyl, halogeno(lower)alkyl, carboxy(lower)alkyl, (lower)alkoxycarbonyl(lower)alkyl, (C6-C o)aryl, (C ₇ -Cn)arylalkyl, di(lower) alkylamino (lower)alkyl and fluoro-substituted analogs of the foregoing, wherein X is -CONR ₂₃ -, -CSNR ₂₃ -, -NR24CO, -NR24CS-, -NR24CONR23-, -NR 4C(NR25)NR23-, -SO2NR23-, -CO, -CSS-, -SCS-, -O, -NR23-, -(NO)R23-, -(PO)R23-, -NR24COO, -NR24SO2-, -S-, -SO or -SO2-, wherein R2 ₃ is selected from the group consisting of hydrogen, lower alkyl and a species which, together with R ₂₁ , forms a saturated or unsaturated heterocydic ring having at least one nitrogen atom and fluoro-substituted analogs of the foregoing, wherein R ₂₄ is hydrogen or lower alkyl and wherein R25 is hydrogen, nitrile or nitro; or XR ₂₁ forms a tetrazole ring; or

wherein the dotted line represents an optional pi bond, wherein R ^* 3, g, R'lO/ A ¹ , A ¹¹ , Y, X, R ₂₁ and u are as defined previously; or

wherein the dotted line represents an optional pi bond, especially in trans configuration, wherein R3, R'10, Rό A ¹ , A ¹¹ , Y, X, R21 and u are defined as previously, wherein R7 is preferably selected from the group consisting of hydrogen, halogen, lower alkyl, amino, nitro, nitroso, nitrile, lower alkylamino and dilower alkylamino; or

XI

wherein the dotted line represents an optional double bond, wherein R3, R'10, A ¹ , A ¹¹ , Y, X, R21 and u are defined as previously for the formula I and VI, wherein A ¹ -[Y-A ¹¹ ] _u XR2i is preferred in α configuration and wherein Z is absent or selected from the group consisting of lower alkylene, halogeno lower alkylene, -(CH ₂ ) _n O, -(CH ₂ ) _n S-, -(CH ₂ )nSe-,-(CH ₂ )nSO-, -(CH ₂ ) _n S0 ₂ -, -(CH ₂ ) _n CO, -(CH2)nNR22-, -(CH2) _n SiR22R22- or -(CH ₂ ) _n R22θR22-, (wherein R22 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

effective amount (discussed supra) of a sex steroid-activity inhibitor as defined above.

Preferred estrogenic nudei suitable for substitution with the -R^f-B-R^xL-G side chain in accordance with the invention indude but are not limited to compounds reported in the literature as having estrogenic activity, natural estrogens such as estradiol, estrogenic derivatives thereof, and other nudei which provide the threshold increase in uterine weight of ovariectomized rats set forth above as defining an estrogenic nudeus (Simard et al., Mol. EndocrinoL 2: 775-784, 1988).

Some preferred estrogenic nudei indude but are not limited to:

wherein x is a halogen, preferably chlorine or iodine;

wherein R ₃ and R ₄ are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C1-C20)/ alkanoyl, (C ₃ -C20) alkenoyl, (C3-C20) alkynoyl and (C7-C11) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherdn R3 is selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C1-C20), alkanoyl, (C3-C20) alkenoyl, (C3-C20) alkynoyl and (C7-C11) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherein R ₃ is selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C ₁ -C20), alkanoyl, (C3-C ₂ Q) alkenoyl,

(C ₃ -C ₂₀ ) alkynoyl and (C ₇ -C ₁₁ ) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherein R3 is sdected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C1-C20)/ alkanoyl, (C3-C20) alkenoyl, (C3-C20) alkynoyl and (C7-C11) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherein R3 is selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C ₁ -C20)/ alkanoyl, (C3-C20) alkenoyl,

(C ₃ -C ₂₀ ), alkynoyl and (C ₇ -C ₁₁ ) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherein the dotted line are an optional double bonds;

wherein R5 and _$ are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C ₁ -C ₂ 0)/ alkanoyl, (C3-C20) alkenoyl, (C3-C20) alkenoyl, (C3-C20) alkynoyl and (C7-C11) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or-

wherein the dotted line is an optional double bond;

whereϊn R5 and R are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C ₁ -C20), alkanoyl, (C3-C2 ₀ ) alkenoyl, (C3-C20) alkynoyl and (C7-C11) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherein the dotted lines are optional double bonds;

wherein R5 and s are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C1-C20), alkanoyl, (C3-C20) alkenoyl, (C3-C20) alkynoyl and (C7-C11) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherein the dotted line is an optional double bond

wherein R5 and R are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C ₁ -C20), alkanoyl, (C3-C20) alkenoyl, (C3-C20) alkynoyl and (C7-C11) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; - or -

wherein q is absent, methylene or ethylene

wherein R5 and are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy carbonyl, (C ₇ -C ₁ 1) aroyl, alkylsilyl, 1-alkyloxy-alkyl and 1-alkyloxy cydoalkyl; -or-

wherein Z is selected from the group consisting of lower alkylene, halogeno lower alkylene, -(CH ₂ ) _n O, -(CH ₂ ) _n S-, -(CH ₂ ) _n Se-, -(CH ₂ ) _n SO-, -(CH2)nSθ2v-(CH2)nCO, -(CH2) _n NR22-, -{CH2) _n SiR22R22- or -(CH2) _n CR22θR22 (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, OR3V ORio', halogen, lower alkyl, lower alkoxy, lower alkoxy carbonyloxy, carboxyl, (C1-C20) alkanoyloxy, (C3-C20) alkenoyloxy (C3-C20) alkynoyloxy, (C7-C11) aroyloxy andalkylsilyloxy;

wherein R'3 and R'ιo are hydrogen, alkyl, (C1-C20) alkanoyl, (C3-C20) alkenoyl, (C3-C20) alkynoyl or (C7-C11) aroyl;

wherein R6 is preferably sdected from the group consisting of hydrogen, nitro, nitrile, halogen, lower alkyl, lower alkenyl, lower alkynyl, halogeno lower alkyl, halogeno lower alkenyl, halogeno lower alkynyl, alkyl sulfonyl, aryl sulfonyl, a substituted 5 to 7 member heterocydic ring having at least one hetero atom (selected from oxygen, sulfur, silicon, selenium, nitrogen), -(CH ₂ ) _S W (wherein W is nitrile, hydroxyl, azido, nitroso, alkoxy, nitro, thionitrile, halogen, alkyl sulfonyl, aryl sulfonyl and s in an integer from 1 to 6), or has 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 Rsi is hydrogen, hydroxyl, halogen, lower alkyl, lower alkenyl, lower alkynyl, nitrile, nitro, nitroso or X ( .2) _n s, wherein Xg is selected from the group consisting -O, -S-, -Se-, -SO, -SO2- and -CO, and Υ 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.

^-[-B-R^lxL-G). Preferred sex steroid activity inhibitors in accordance with the invention indude are not limited to:

N-n-butyl-N-methyl-ll-(16 ^, α-bromo-3 ^, ,17 ^, β-dihydroxy-estra-l ^, ^'^'(10 ^, )-trien-7 ^* α-yl) undecanamide ("EM 105"):

N- -butyl-N-met yl-ll-(16 ^, o-b^omo-3 ^, ,17 ^, α-dihyd^oxy-estra-l ^, ^ ^{, ,} 0 ^, )-trien-7 ^, α-yl) undecanamide ("EM 171"):

N-n-butyl-N-methyl-ll-(16 ^, α-chloro-3 ^, ,17 ^• β-dihydroxy-estra-l ^, ^ ^, ^ ^• (10 ^, )-tτien-7 ^, α-yl) undecanamide ("EM 139"):

N-n-butyl-N-me yl-ll-(16'α-chloro-3 ^, ,1 α-dihyd^oxy-estra-l'^ ^, ^ ^• (10 ^, )-trien-7 ^, α-yl)undecanamide ("EM 170"):

N-n-butyl-N-methyl-ll-(16 ^, α-iodo-3',17'β-dihydroxy-estra-l ^, ^ ^, ^ ^, (10 ^, )-trien-7 ^, α- yl) undecanamide ("EM 156"):

N-n-butyl-N-methyl-ll-O'-hydroxy-l^-oxo-estra-l^^'dO _/ lS'-tetraen-^α-yl) undecanamide ("EM 112"):

N-n-butyl-N-methyl-ll-OM^β-dihydroxy-l^α-ethynyl-estra- l^^'dO _/ lS'-tetr aen-7'α-yl)undecanamide ("EM 123"):

N-n-butyl-N-methyl-ll-(3 ^, ,17 ^, β-dihydroxy-17 ^, α-ethynyl-estra-l ^, ,3'^ ^, (10 ^, ),14 ^, -tetr aen-7O-yl)undecanamide ("EM 140"):

N-n-butyl-N-methyHl-(3 ^, ,17 ^, β-dihyd^oxy-15'β,16 ^, β-methylene-estra-l ^, ^ ^, ,5'(10 ^, ), lS'-trien^'α-yDundecanamide ("EM 136"):

N-n-butyl-N-methyl-ll-(3 ^, ,17'β-dihydroxy-17'α-ethynyl-estra-15 ^, β,16 ^, β- methylene-estra-l'^' _/ S'dO -trien-^α-yDundecanamide CΕM 138"):

N-n-butyl-N-methyl-ll-(3 ^, -hydroxy-15'β,16 ^, β-methylene-17'-oxo-estra- l',3 ^, ,5 ^, (10 ^, )-trien-7 ^, α-yl)undecanamide ("EM 137"):

N-n-butyl-N-methyl-ll-O'-hydroxy-lό'-methylene-l^-oxo-es tra-l'^'^'dO')- trien-7'α-yl) undecanamide ("EM 175"):

N-n-butyl-N-methyl-ll-(3 ^, ,17 ^, β-dibenzoyl-14 ^, β,15 ^, β-epoxy-estra-l ^, ^ ^, ^ ^, (10')- trien-TO-yDundecanamide (ΕM 180"):

N-n-butyl-N-methyl-ll-(3 ^, ,17 ^, β-dibenzoyl-14 ^, o,15 ^, α-epoxy-estra-l ^, 3 ^, ^ ^, (10 ^* )- trien-7α-yl)undecanamide ("EM 181"):

N-n-butyl-N-methyl-ll-(3 ^, ,17 ^, β-dihydroxy-estra-l'^ ^, ^ ^, (10 ^, )-trien-7 ^, α- yDundecanamide ("EM 108"):

N-n-butyl-N-methyl-13-(3',17 ^, β-dihydroxy-estra-l'^ ^, ^ ^, (10 ^, )-trien- α-yl)12- tridecynamide ("EM 163"):

C≡C(CH ₂ ) ₁₀ CON(CH ₃ )C ₄ H ₉

N-n-butyl-N-methyl-14-(31 ihyd^o y-estra-l ^, ^ ^, ^ ^, (10 ^, )-trien- α-yl)13- tetradecynamide ("EM 195"):

N-n-butyi-N-methyl-8-(3 ^, ,17 ^, β-dihydroxy-estra-l ^, ^ ^, ^ ^, (10')-trien- α-yl)7- octynamide ("EM 157"):

N-n-butyl-N-methyl-ll-(6 ^, -hydroxy-2 ^, -(4"-hydroxyphenyl)-3'ethyl-indol-N'-yl) undecanamide (EM 215):

(CH ₂ ) ₁₀ CON(CH ₃ )C ₄ H ₉

N-n-butyl-N-methyl-ll-(6'-hydroxy-2 ^, -(4"-hyd^oxyphenyl)-3 ^, ,4 ^, - dihydronaphtalene-3'-yl) undecanamide:

N-n-butyl-N-methyl-ll-^ '-d^-diethyl-l^-ethanydyD bis-phenol-S-yl)] undecanamide (EM 406):

6-hydro)cy-2-(4 ^, -hydroxyphenyl)-l-methyl-3-(6"-piperidino)hexyl-3,4- dihydronaphatalene (EM 473)

N-n-butyl-N-methyl-ll-(6'-hydroxy-2 ^, -(4 ^,, -hydroxyphenyl)-l ^, -methyl-3 ^, ,4'- dihydronaphatalene-3'-yl) undecanamide (EM 690)

EM 690

6-hydroxy-2-(4'-hydroxyphenyl)-l-methyl-3-pentafluoropentyls ulphinylnonyl- 3,4-dihydronaphatalene (EM 732)

2-(4 ^, -hydroxyphenyl)-l-methyl-3-[4"-(2 ^, "-piperidinoethoxy)phenyl]-3,4- dihydronaphatalene (EM 765)

2-(4'-hydroxyphenyl)-l-methyl-3-[4"-(2 ^, "-piperidinoethoxy)benzyl]-3,4- dihydronaphatalene (EM 431)

N-pyrrolidinyl-ll-(7'-hydroxy-3 ^, -(4 ^,, -hydroxyphenyl)-4 ^, -methyl-2H- benzothiopyran-2 -yl) undecanamide (EM 941)

7-hydroxy-3-(4'-hydro3yphenyl)-4-methyl-2-[3-(2"-piperidi no)ethoxy]propynyi- 2H-benzopyran (EM 555)

N-n-butyi-N-methyl-ll-(7 ^, -hydroxy-3 ^, -(4"-hydroxyphenyl)-4 ^, -methyl-2H- benzopyran-2-yl) undecanamide (EM 467)

7-hydroxy-3-(4 ^, -hydroxyphenyl)-4-methyl-2-(6"-piperidino)hexyl-2H- benzopyran (EM 721)

When a sex steroid nudeus is substituted with the side chain R ¹ [-B-R ^2_ ] _x L-G, it is preferred that the side chain have between about 7 and 30 carbon atoms and that L be separated from the nudeus by at least 3 intervening and preferably 6 atoms. In some embodiments, a polar moiety (G, L or both) is preferably separated from the nudeus by at least 8 intervening atoms.

Additional inhibitors of the formula:

Indude but are not limited to those set forth below:

The following compound of the invention:

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.

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 add chloride 1 (20.0g; O.lmol) was added dropwise to methanol (60ml) at room temperature and with stirring. The solution was refluxed for lh. 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 (MgSCλt) 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%).

The above ester 2 (21.2g; 0.108mol) and the nitrile 3 (available from Aldrich Chemical Company Inc., Milwaukee, Wis.) (175ml; 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 IN hydrochloric add aqueous solution. The organic solution was dried (MgSC» 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; 69%). (δ NMR; 300MHz; solvent: CDC1 ₃ ; standard: TMS) 3.78 (3H; s; OMe) 3.84 (3H; s; OMe) 3.95 (3H; s; OMe) 5.84 (1H; s; 0=C-CH-CN) 6.43 (1H; d; J 2.5Hz; CH phenyl) 654 (1H; dd; J 2.5Hz and 85Hz; CH phenyl) 6.89 and 7.35 (2H; AB system; J 85Hz; CH phenyl) 7.79 (1H; d; J 85Hz; CH phenyl) .

A solution of the ketone 4 (37.8g; 0.12mol) in acetic add (400ml) and concentrated hydrochloric add aqueous solution (200ml) was kept at 90°C for 3h. The reaction mixture was neutralized with concentrated sodium hydroxide aqueous solution and it was extracted with ethyl acetate. The organic extrart was dried (MgSθ 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; 20%).

Triphenol 23

To a mbcture of the ketones 5 (17.0g; 59.4mmol) and 6 (6.6g; 243mmol) was added pyridine hydrochloride (90g). The mixture was heated at 220°C for 20min. IN hydrodiloric add aqueous solution (250ml) was added and the resulting mixture was extracted several times with ethyl acetate. The organic extract was dried MgSθ ₄ ) 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).

Diether24

To a mixture of the triphenol 23 (14-lg; 57.8mmol) in 3,4-dihydro-2H-pyran (200ml) at 0°C and with vigourous stirring was carefully added p- toluenesulfonic add monohydrate (2.0g). 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 (MgSC» 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; 98%). (δ NMR 300MHz; solvent CDC1 ₃ ; standard: TMS) 1.5-2.1 (12H; m; C 2H-CH ₂ -CH ₂ -CH ₂ -CH ₂ -0 THP) 355-3.65 (2H; m; 0-CH-CH ₂ - CH ₂ -CH ₂ -CH ₂ -O THP) 3.75-3.95 (2H; ; O-CH-CH ₂ -CH ₂ -CH ₂ -CH ₂ -O THP) 4.16

(2H; s; Ph-CH ₂ -C=0) 5.40 (IH; t; J 3Hz; 0-CH-CH ₂ -CH ₂ -CH ₂ -CH ₂ -0 THP) 5.49 (IH; t; J 3Hz; OCH-CH ₂ -CH ₂ -CH ₂ -CH ₂ -O THP) 655 (IH; dd; J 25Hz and 85Hz; CH phenyl) 6.61 (IH; d; J 2.5Hz; CH phenyl) 7.03 and 7.17 (2H; AB system; J 85Hz; CH phenyl) 7.77 (IH; d; J 8.5Hz; CH phenyl) 12.60 (IH; s; Ph-OH).

Ghroman-4-one 25 (R=H) and Chalcone 26 (R=H)

To a mixture of the diether 24 (24.4g; 59.2mmol) and the aldehyde (OHCC-6H4OH) (7.6g; 62.18mmol) in dry benzene (750ml) was added piperidine (500μl). 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 (R=H) (14.3g; 47%) and 26 (R=H) (8.4g; 27%). This last compound can be converted to compound 25 (R=H) by heating with sodium acetate in methanol.

Chroman-4-one 25 (R=H)

(6 NMR 300MHz; solvent: CDCI3; 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 - H ₂ -CH ₂ -CH ₂ -O THP) 4.05-4.1

(IH; m 0-CH-CH-C=0) 5.25-5.35 (IH; m 0-CH-CH-C=0) 5.35-555 (2H; m; O-CH-

CH ₂ -CH ₂ -CH ₂ -CH ₂ -O THP) 6.6-7.1 (10H; m; CH phenyl) 7.85-7.95 (IH; m; CH phenyl).

66-

SCHEME

EM 343

A mixture of compound 25 (R=H) (1.90g; 3.8mmol), l-(2-chloroethyl)piperidine hydrochloride Cl(CH _{2 2} NC ₅ Hι ₀ . HCl (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 predpitate 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 (157g; 66%). To a solution of compound 25 (R CH _∑ NCsHio) (90mg; 143μmol) in ether (30ml) 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 (MgSC> ₄ ) 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 , Re = CH )(90mg; 97%) which was used directly in the next reaction.

A solution of compound 27 Re = CH ₃ ) (90mg; 139μmol) in a mixture of acetic add (60ml) and water (6ml) was kept at 100°C for lOmin. 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 NC ₅ Hιo , Re = CH ₃ ) (40mg; 62%). (δ NMR; 300MHz; solvent: CP ₃ OD; standard: TMS), 1.46 (2H; m; cydo-N-CH ₂ -CH ₂ -CH ₂ -CH ₂ -CH ₂ ) 1.60 (4H;

m; cydo-N-CH ₂ -CH ₂ -CH ₂ -CH ₂ -CH ₂ -) 2.02 (3H; s; CH ₃ -C=C) 2.56 (4H; m; cyclo-N- CH ₂ -CH ₂ -CH ₂ -CH ₂ -CH ₂ -) 2.76 (2H; r; J 5Hz, O-CH ₂ -CH ₂ -N) 4.06 (2H; t; J 5Hz; O- CH ₂ -CH ₂ -N) 5.77 (IH; s; O-CH-Ph) 6.12 (IH; d; J 25Hz; CH Phenyl) 655 (IH; dd; J 25Hz,8Hz; CH Phenyl) 6.70 (2H; d; J 85Hz; CH Phenyl) 6.77 (2H; d; J 85Hz; CH Phenyl) 6.98 (2H; d; J 8.5Hz; CH Phenyl) 7.12 (IH d; J 8Hz; CH Phenyl) 7.19 (2H; d; J 85Hz; CH Phenyl). Mass Spectroscopy: M+ 459.

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

Maintenance of Stock Cell Cultures

ZR-75-1 cells (83rd passage) were obtained from the American Type Culture Collection (Rockville, MD) and routinely cultured in phenol-red free RPMI 1640 supplemented with 1 nM E2, 2 mM L-glutamine, 1 mM sodium pyruvate, 15 mM N-2-hydroxyethyl-piperazine-ZsT'-2-ethanesulfonic acid, 100 IU penidllin/ml, 100 μg streptomyrin/ml, and 10% (v/v) fetal bovine serum (Hydone, Logan, UT) under a humidified atmosphere of 95% air, 5% CO2, at 37 °C All media and medium supplements were purdiased from Sigma. Cells were subcultured weekly by treatment with a pancreatic solution containing 0.02% EDTA (w/v). The cell cultures used for the experiments herein described were between passages 89 and 94.

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 IO ⁴ cells /well. After 72 h, medium was replaced with fresh medium of identical composition containing, in addition, the concentrations of steroids and/or inhibitors (e.g. EM 312 or EM 343) indicated along the X-axis of Hgure 5. Control cultures received the ethanol vehide only. Cells were then allowed to grow at 37°C for 10 days with medium dranges (of identical composition) every 2 days. In absence of inhibitors, in 0.1 nM estradiol (E ₂ )-containing medium, ZR-75-1 cells have doubling time of about 48 h.

After E2 and/or antiestrogen treatment, cells were harvested by addition of 0.5 ml of a pancreatin solution (Sigma) for 5-10 min at 37 °C before addition of 0.5 ml of RPMI 1640 containing 5% dextran-coated charcoal-fetal bovine serum in order to block enzymatic action. Cell number (0.10-ml 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 255 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 (e.g. the methyl substitution of EM 343) enhances effectiveness relative to compounds lacking such a substitution. For example, another spedes of the invention.

reported by Saeed et al (J. Med. Chem. 33: 3210-3216, 1990) and Shaπna et al. (J. Med. Chem. 33: 3216-3222 and 3222-3229, 1990) to have antiestrogenic activity underperformed EM 343 in comparative testing in our laboratory on the growth of human ZR-75-1 breast cancer cells, the IC50 value of EM-312 being 3-fold higher at 8.43 x 10-lθM (Fig.5).

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

pathways and variations capable of producing a variety of antiestrogens and other sex steroid activity inhibitors in accordance with the invention.

EXAMPLES OF SYNTHESIS OF PREFERRED INHIBITORS OF SEX STEROID ACTTVITY

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 δ 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 Kiesdgel 60F254 plates (E. Merck, Darmstadt, FRG). For flash chromatography, Merck-Kieselgd 60 (230-400 mesh A.S.T.M.) was used. All solvents used in chromatography have been distilled. Unless otherwise noted, starting material and reactant were obtained commerάally 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.

IIST OF ABBREVIATIONS

Bz Benzyl

DMF Dimethylformamide

EDTA Ethylenediaminetretraacetic add

HMPA Hexamethylphosphoramide

HPLC High pressure liquid chromatography

LDA Lithium diisopropylamine mCPBA meta-chloroperbenzoic add

MOM Methyloxymethyl

NAD Nicotinamide Adenine Dinudeotide

NADH Nicotinamide Adenine Dinudeotide reduced form

PTSA Para-toluene sulfonic add

PTSCl Para-toluene sulfonyl chloride

TBDMS t-butyldimethylsilyl

THF Tetrahydrofuran

THP Tetrahydropyrannyl

TMS Tetramethylsilyl

EXAMPLE I

Instrumentation

IR spectra were obtained in a Perkin-Elmer spectrometer 1310. UN spectra were recorded in methanol on a Beckman DU-6 spectrometer. H-ΝMR spectra were obtained at 200 MHz 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.

Ν-buty Ν-methyl-12,13-Bis (4-hydroxyphenyl)-12-pentadecenoic amide (EM-142, compound 5 with x=10)

The synthesis of this compound is described in the scheme I (infra) where x=10.

12,13-Bis (4-methoxyphenyl)-ll-pentadecenol (3)

4'-methoxy-2-ethyl,2-(4-methoxyphenyl). acetophenone (2) (710 mg, 2.5 mmol, prepared from desoxyanisoin, ethyl bromide and LDA by a known method) in THF (10 ml) were added, under argon, to Grignard reagent prepared from 11-bromo-tetrahydropyranyl undecanol (6.6 g, 19.7 mmoles) and magnesium

(0.6 g, 24.7 mmoles) and THF (10 ml). The mixture was stirred for 18 hours, then addified with IN HCl and extracted three times with ether. The organic phase was washed with water (X3), dried over anhydrous magnesium sulfate, and evaporated under reduced pressure. The residue was chromatographed on Silica-gd (Kieselgel, 60F254, Merck, 0.063-0.200 mm, 100 g). Elution with a mixture of hexane-ethyl acetate (9:1 v/v) gave 12,13-Bis-(4-methoxyphenyl)- tetrahydropyranyl pentadecan-l,12-diol (991 mg, 76%) as a mixture of diastereoisomers; colorless oil, IRv _ma χ (neat) 3480, 1600 cm ^*1 ; 'H-NMR (δ, CDC1 ₃ ); 0.62 (3H, t, J= 75 Hz, CH2CH ₃ ), 2.73 (IH, 2d, J=9.7 Hz, -CHCH ₂ CH ₃ ), 3.254.00 (4H, m, -CH2OCHOCH2-), 3.76 and3.79 (6H, 2s, -OCH3), 4.57 (IH, t, J=l.l Hz, -O2-CH-CH2) and 6.71-730 (8H, m,H-Ar)ppm. MS m/e= 523 (M+-H ₂ 0).

The above diastereoisomers (920 mg, 1.8 mmol) dissolved in methanol (30 ml) and 5N HCl (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 g). Elution was a mixture of hexane-ethyl acetate (7:3 v/v) gave 12,13-Bis(4-methoxyphenyl)-ll-pentadecenol (3) (710 mg, 65% from compound 2), colorless oil, IR, v _max (neat), 3340, 1600, 1030 on" ¹ ; UV γ _max (log ε) = 231

(4.27) nm; 'H-NMR (δ, CDCI ₃ ), 0.88 (3H, t, J = 7.3 Hz, -CH ₂ CH ₃ ), 3.30 (IH, t, J= 9.7 Hz, -CH-CH ₂ ,CH ₃ ) 3.63 (2H, t, J = 6.6 Hz, -CH2OH), 3.76 and 3.78 (6H, 2s, -OCH ₃ ),

551 (IH, t, J= 8.8 Hz, 1 -C = CH-) and 6.63-7.108H, m, H-Ar) ppm; MS m/e = 438 (M+).

N-butyl. N-methyL12.13-Bis (4-methoxyphenyl)-ll-pentadecenoic amide (4)

To a cooled solution of alcohol 3 (710 mg, 1.56 mmol) in acetone (17 ml) was added Jones' reagent (8N-chromic add 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 12,13-Bis (4-methoxyphenyl)-ll-pantadecenoic add 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 μL 1.96 mmol) and isobutylchloroformate (280 μl, 2.1 mmol). After 40 minutes, N-methylbutylamine (15 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 HCl, saturated sodium bicarbonate solution and water (3X), 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 g). Elution with a mixture of hexane-ethyl acetate (4:1 v/v) gave N-butyl, N-methyl-12,13-Bis

(4-methoxyphenyl)-ll-pentadecenoic amide (4), (549 mg, 68%) colorless oil; IR v _max (neat), 1640, 1600 cm- ^l ; UN γ _max Oog ε) = 230, (459) nm; ⁱ H-ΝMR (δ,

CDC1 ₃ ), 0.85-0.98 (6H, m, 2-CH2CH3), 2.27 (2H, t, J= 7.1 Hz, CH2COΝ), 2.91 and 2.96 (3H, 2s, -NCH ₃ ), 3.25-356 (3H, m, -NCH2 and CH ₃ CH2CH-), 3.77 and 3.78 (6H, 2s, OCH ), 550 (IH, J= 7.1 Hz, -C=CH-) and 6.69-7.01 (8H, m, H-Ar) ppm; MS m/e = 521 (M+).

N-butyL N-methyl-12.13-Bis (4-hydroxyphenyl)-12-pentadecenoic amide (EM-142, compound 5 with x = 10).

To the above dimethoxy amide 4 (117 mg, 0.22 mmol) in CH2CI2 (1 ml) at 0°C was added, under argon, 1.0 M borane tribromide (675 μl). The solution was stirred for 1 hour, then poured into water and extracted with ether (3x). The organic solution was washed with water, dried on magnesium sulfate, and evaporated to dryness. The residue was purified by "Hash chromatography" on silica gel (Kieselgel 60, Merck, under 0.063 mm, 30 g). 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 = 10) (34 mg, 31%), colorless oil, IR v _max (neat) 3300, 1600 cm- ¹ ; UN γ _max (log ε) =

235 (4.25) nm; *H-ΝMR (δ, CDCI ₃ ), 0.76 (3H, t, J= 7.3 Hz, -CH ₂ CH3), 0.96 (3H, t, J= 7.3 Hz, N (CH ₂ ) ₃ CH ₃ ), 2.05-2.20 (4H, m, CH _∑ -C = C-CH ₂ -), 2.35 (2H, t, J= 7.0 Hz,

-CH ₂ CON-), 2.97 and 3.00 (3H, s, -NCH3), 3.29 and 3.41 (2H, 2t, J= 73 Hz, -N-CH2-), and 659-7.09 (8H, m, H-Ar) ppm; MS m/e = 493 (M+).

Sdieme 1

(CH2)x ₊ ιOTHP

5

EXAMPLE2

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 Assdin 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°C for 3 hours). It can be seen that EM-142 is only 3 times less potent than 17β-estradiol itself while being more potent than the antiestrogen IO 164384.

The antiestrogenic activity of EM-142 was measured in vivo by inhibition of 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-142, and/or estradiol contained in ethanol were injected subcutaneously in the appropriate groups in a solution of 0.9% (w/v) sodium chloride and 1% (w/v) 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 μg 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

9-10 mice. It can be seen that the very low dose of 05 μg already has a significant inhibitory effect of an E ₂ - ^induced uterine growth and that a complete reversal of E2 effect is obtained at higher doses. A half-maximal inhibitory effect is in fact observed at approximately 3 μg while, at the doses used, 20 μg causes a complete reversal of estrogenic action. Note that EM-142 has no estrogenic effect on uterine weight, thus demonstrating its potent antiestrogenic activity.

- 82 -

EXAMPLE3 Scheme 2

N-buty N-methyl-12,13-Bis-(4-hydroxyphenyl ⁾ pentadecanoic amide (6)

EXAMPLE 4 Scheme 3

N-butyl, N-methyl-5-f4-f(2-[4-hydroxyphenyl. benzothiophen-3-yl) formyll phenoxyi hexanoic amide (12)

f^dCOCeH.OCHzCsHs

Aids

EXAMPLE 5 (Scheme 4)

N-butyl, N-methyl-6-fp-(trans-l' ,2'-bis (4"-hvdroxyphenyl)-l'-butenyl) phenoxyll heptanoic amide (18)

18

EXAMPLE 6 Scheme 5

N-butyl. N-methyl-(6'-hvdroxy, 2'-(4"-hvdroxyphenyl)-3'-methyl-indoI-N'-yl) undecanoic amide (22)

EXAMPLE 7 Scheme 6

N-butyl, N-Methyl-1 l-fe '-Q ,2-diethyl-l 2-ethanydiyl) bis-phenol-3-yn undecanoic amide (28)

EXAMPLE 8 (Scheme 7) N-butyl, N-methyl- 6'-hydroxy-2'-(4-hydroxyphenyl)-(T 2'-dihydronaphta-len- 3'-yl) undecanoic amide (34)

6-methoxy-2-tetralone 29

NaH RBr (1 eq)

34

EXAMPLE9

SYNTHESIS OF A STARTING COMPOUND, N-N-BUTYL, N-METHYL-ll-(3'- BENZOYLOXY-17 ^, -OXO-ESTRA-l ^, ,3',5 ^, (10 ^* )-TRIEN-7 ^l α-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 men 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%) p. 176-177°C The structure was confirmed by spectroscopic means.

176-acetoxy-estra-4.6-dien-3-one (3)

To a cooled suspension of enolacetate (121 g; 0.337 mole) in a mixture of DMF (330 ml) and water. (7.2 ml) at 0°C was added, under nitrogen, over a period of 1 h, N-bromosuccinimide (63 g). The resulting solution was stirred for an

additional 0.5 h at 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 1.71 of ice-cold water containing 165 ml of glacial acetic acid. After stirring during 15 hours, the crude 17β-acetoxy-estra-4,6-dien-3-one (3) was filtered, washed with water, dried in a desiccating apparatus and recrystallized twice from isopropyl ether (72 g, yield 68%, mp 110°C). The structure was confirmed by spectroscopic means.

7α-(ll'-acetoxy-undecyl) 17β-acetoxy estra-4-en-3-one (4)

A. Preparation of reagents and solvents

11-bromo undecanol tetrahydro pyranyl ether

11-bromo-undecanol (100 g, 398 mmol) was dissolved in dry ether (768 ml) and the solution was cooled to 0°C using an ice/H _∑ O bath. To this solution was added HCl gas (2.13 g, 58.4 mmol, 26 ml of HCl/ether).

To this mixture, a solution of 3,4-dihydro-2H-pyran (39.9 g, 43.3 ml) freshly distilled in dry ether (218 ml) was added over a period of 90 min. The solution 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. Grignard reagent

Li 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°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 -20°C

The mixture was cooled to -35°C C£2°C) using a dry ice/CCU/acetone bath. The anhydrous cuprous chloride (1.18 g, 12 mmol) was added and the mixture was stirred over a period of 0.5h.

C. Addition of Grignard reagent

After 0.5 h, using the same apparatus mentioned above (Ar, -35°C), a solution of 17β-acetoxy estra-4,6-diene-3-one (3) (32.0 g, 102 mmol) 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), diluted with water and extracted with ether (3x). 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 HCl (180 ml), refluxed for 1 h and 45 min, then concentrated under reduced pressure and cooled in an ice bath. The mixture was then filtered to remove the white precipitate. After the solution had been diluted with water and extracted with methylene chloride (3x), the organic layer was dried over anhydrous MgSC* ₄ 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 7α-(ll'-hydroxy-undecyl)-17β-hydroxy estra-4-en-3-one (34.8 g) which was dissolved in dry pyridine (200 ml) and dry

acetic anhydride (200 ml), stirred 17h at room temperature and then poured in ice-water. The product was extracted with methylene chloride (3X), 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 7β-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 v/v). The first product eluted was pure amorphous 7α-(H'-acetoxy undecyl) 17β-acetθ3cy-estrar4-en-3-one (4) (20.8 g, 39.4 mmol, yield from dienone was 39.0%). Further elution gave the 7β-isomer (5) (5.4 g, 103 mmol, 10%). All structures were determined by spectroscopic means.

7α-(ll'-hydroxy-undecyl) estra-1 3(10)-trien-3.17B-diol (6a)

Under dry argon, a solution of 7α-(ll'-acetoxy undecyl) 17β-acetoxy-estra-4-en-3-one (4) (17.0 g, 32.4 mmol) in dry acetonitriie (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 acetonitriie (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

compound was extracted with ethyl acetate (3x 150 ml). 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 mm; 1000 g). Elution with hexane-ethyl acetate (1:1 v/v) gave the 7α-(ll'-acetoxy-undecyl) estra- l ^, ,3',5'(10 ^, )-trien-3,17β-diol, 17β-acetate (6b) (8.51 g; 503%) and the starting product (133 g; 15%).

The above diacetate phenol (851 g, 16.2 mmol) was dissolved in methanol (90 ml) and sodium hydroxyde 30% (w/v) (9 ml). The mixture was refluxed for 90 min under dry nitrogen. The solution was then concentrated under vacuum and diluted with hydrochloric acid (10%, v/v). The mixture was extracted using ethyl acetate (4 x 150 ml) and the ethyl acetate extract was washed with water, dried over anhydrous magnesium sulfate, filtered and evaporated under vacuum. The evaporation gave 7α -(ll '-hydroxy undecyl) estra-l,3,5(10)-trien-3,17β-diol (6a) (6.99 g, 98% brut) as a yellow foam, the structure of which was confirmed by spectroscopic means.

3-benzoyloxy 7α-(ll'-hvdroxy undecyl) estra-1 .5(10)-tτien-17&-ol (7)

The above triol (6.99 g; 15.8 mol) was dissolved in acetone (25 ml) and an aqueous solution of sodium hydroxyde ( N, 19.1 ml). The mixture was cooled

to 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 0°C 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 g). The chromatography was carried out, first, using methylene chloride as solvent (about 1 liter) and secondly the pure 3-benzoyloxy 7α-(ir-hydroxy undecyl) estra-13,5(10)-trien-17β-ol (7), colorless oil (630 g, 75%) was eluted with methylene chloride-ethyl acetate (5:1 about 1 liter and 4:1; v/v). The structure was confirmed by spectroscopic means.

ll-(3-benzoyloxy-17 ^/ -oxo-estra-l ^/ 3 ^/ 5 ^, (10)-trien-7 ^, α-yl) undecanoic acid (8)

To a cooled solution of 3-benzoyloxy-7α-(ll ^, -hydroxy undecyl)estra-l,3,5(10)- trien-17β-ol (7) (4.3 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 ll-(3 ^, -benzoyloxy-17 ^, -oxo-estra-l ^, 3',5'(10 ^/ )-trien-7'α-yl) undecanoic add (8) (3.94 g) was used in the next step without purification.

96-

Scheme 8

9aR _a =C ₆ H _s CO 9b R _a =H

N-n-butyl,n-methyl-ll-(3 ^, -hvdroxy-17 ^/ -oxo-estra-l ^, 3 ^, 5 ^/ (10 ^/ )-trien-7 ^/ α-yl) undecanamide (9b)

To ll-(3'-benzoyloxy-17 ^, -oxo-estra-l',3',5'(10 ^, )-trien-7'α-yl) undecanoic add (8) (3.94 g, 722 mmol), dissolved in anhydrous CH ₂ CI ₂ (100 ml) and cooled at -10°C was added tributylamine (2.18 ml, 9.15 mmol) and isobutylchloroformate (1.30 ml, 10.0 mmol). The solution was stirred during 35 min. and N- methylbutylamine (13 ml, 109.7 mmol) was added. The mixture was warmed to room temperature and stirred during 1 h. Afterward, CH2CI2 was added and the organic phase was washed with IN HCl, water, saturated sodium bicarbonate solution and finally with water, dried with anhydrous MgSθ4 and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel. Elution with mixture of EtOAc/hexane (15:85 v/v) yielded N-butyl, N-methyl-11-(3'- benzoyloxy-17'-oxo-estra-l'3',5'(10')- trien-7'α-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, 054 mmol) was dissolved in methanol (10 ml) and cooled at 0°C. Following this 2N NaOH (5 ml) was added and the mixture was stirred during 60 min. at 0°C The solution was neutralized with IN HCl and extracted with CH2CI2. The organic phase was dried with anhydrous MgSθ 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-1 l-(3'-hydroxy-l 7'-oxo,estra-l ',3',4' (10)-trien-7' α-yl) undecanamide (9b) (284 mg, 97%) as colorless oil; ^J H-NMR δ (CDCI3) 0.91 (s3H,18'-CH ₃ ), 2.76

app(d,lH,J=I6,3Hz, part of ABX system, 6'-H) 2.96 and 2.98 (2s3H,N-CH ₃ ), 3.27 and 338 (2t _aP p,2H,J=75Hz,N-CH ₂ -) 16.63 (broad s,lH,4'-H), 6.70 (broad d,lH,J=8.5 Hz,2'-H), 7.12 (d,lH,J=8.4 Hz,l'-H); IRVmax (neat) 3270, 1730, 1615 cm- ¹ ; MS m/e 523 (M+,100%), 508 (M+-CH3 2%), 142 (C2H CO (CH3)C H9 ⁺ , 47%).

16-HALO-ESTRADIOL UNDECANAMIDE (SCHEME 9)

N-n-butyLN-methyl-ll-(3 ^, .17 ^, β-diacetoxy~estra-l ^, 3 ^, 5 ^, (10 ^, ).16 ^/ -tetraen-7α-yl) undecanamide (10)

The ketone amide 9b (163 mg, 050 mmol) was dissolved in isoprenyl acetate (10 ml), p-toluenesulfonic add (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 MgS04 and the solvent was removed under reduced pressure. The residue was filtered through alumina (15mm x 50 mm alumina Woehlm neutral, activity II) using a mixture of benzene-diethyl ether (3:7 v/v/) 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-ll-(3',17'β-diacetoxy- estra-l',3',5'(10'), 16'-tetraen-7'α-yl) undecanamide (10) (244 mg, 80%) as colorless oil; ϊ H-NMR δ (CDCl ₃ ) 0.92 (s3H,18'-CH ₃ ), 0.92 and 0.95 (2t3H-3',J=7.0 Hz,N(CH ) ₃ CH ₃ ), 2.18 (s,3H,17'-OCOCH ₃ ), 2.28(s,3H ^/ -OCOCH ₃ ), 2.76 app (d,lH,J=16.1 Hz, part of ABX system,6'-H), 2.90 and 2.96 (2s H,N-CH ₃ ), 3.26 and 3.35 (2tap _P ,2H,J=7.6 Hz,N-CH ₂ -), 5.52 (m,lH,16'-H), 6.80 (broad s,lH,4'-H), 6.85 (dd,lH,Jι=9.1 Hz and J ₂ =3.0 Hz,2'-H), 7.27 (d,lH =9.1 Hz,l'-H); IR v _max (neat)

1750, 1635, 1200 cm ¹ ; MS m/e 607 (M+,2%) 5(M ⁺ -COCH ₂ ,100%), 550 (M+-COCH ₂ -CH ₃ ,13%), 523 (M+-2COCH ₂ 45%), 142 (C ₂ H CON(CH ₃ )C H9 ⁺ 55%), 129 (C4H ₉ (CH ₃ )NCOCH ₃ ⁺ 38%), 114 (C ₄ H ₉ (CH ₃ )NCO ⁺ , 60%), 86 (C4H ₉ (CH ₃ )N ⁺ , 25%); EXACT MASS calcd for C ₃ 8H5 ₇ O5 607.4239. found 607.4234.

N-buty N-memyl-ll-(16 ^/ α-chloro-3 ^, acetoxy-17 ^/ -oxo-estτa-l ^# 3 ^/ .4 ^, (10 ^, )-triene-7 ^, α- yl) undecanamide (11. X = Cl)

To diacetate amide 10, dissolved in 5 ml of acetone, was added a solution of sodium acetate (2.6 equivalents) in acetic add and water (1:113 v/v) and then, was treated with tertbutyl hypochlorite (1 eq.) prepared from t-butanol (4 ml) and Javel water (Javex 6.1%. 50 ml). The dear 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 ml) and water was added (20 ml). The organic phase was washed with water, dried with anhydrous MgSθ4 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-11- (16 ^/ α-chloro-3'-acetoxy-17 ^/ -oxo-estra-l ^, ,3 ^, ,4'(10')-trien-7 ^/ α-yl) undecanamide 11, X=C1) (115 mg, 89%) as colorless oil; Η-NMR δ (CDC1 ₃ ) 0.92 and 0.95 (2t,3H =7.0 Hz,N(CH ₂ ) ₃ CH ₃ ), 0.96 (s3H,18'-CH ₃ ), 2.28 (s,3H,3'-OCOCH ₃ ), 2.80 app (d,lH =16,6 Hz, part of ABX system, 6'-H) 2.90 and 2.96 (2s3H _/ N-CH ₃ ), 3.24 and

3.35 (2tap _P ,2HJ=7.4 Hz,-N-CH ₂ -), 4.46 (d,lH,J=6.6 Hz,16'β-H), 6.82 (broad s,lH,4'-H), 6.86 (dd,lH,J=9.1Hz and T ₂ =,2.6Hz,2'-H), 7.29 (d,lH,J=9.1 Hz,l'-H); IR Vmax (neat 1750, 1640, 1205 cm- ¹ ; MS m/e 601, 599 (M ⁺ ,24%, 68%), 142 (C ₂ H4CON(CH ₃ )C ₄ H9 ⁺ , 100%), 114 (C ₄ H9(CH ₃ )NCO ⁺ , 93%).

N-but l.N-me yl-ll- 16 ^/ α-chloro-3 17 ^, -dihydroxy-esfra-l 3 ^/ 5 ^/ l(r -t^ien-7 ^, α-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°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 0°C for 5 min, then was quenched by the dropwise addition of a mixture of THF-EtOAc (5 ml) (1:1 v/v) and addified at pH ~ 4 with (10%) HCl. 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 Sθ4 and evaporated under reduced pressure. The residue was chromatographed on silica gel with a mixture of EtOAc/hexane (4:6 v/v) as eluant:

N-but L N-methyl-ll- 16 ^/ α-cMoro-3 17 ^/ α-dihydrox -est^a-l ^{, ,} 5 ^/ 10 ^/ -trien-7 ^, α- yl) undecanamide ("EM 170")

(15 mg, 29%) as colorless oil; analytical sample was obtained by HPLC purification; Η-NMR δ (CDC1 ₃ , 400 MHz) 0.79 (s,3H,18'-CH ₃ ), 0.93 and 0.96 (2t, 3H,J=73 Hz,N(CH2)3CH ₃ ), 2.80 (2H,J^6=17.1 Hz and J ₆ ,7 = 45 Hz, Δδ=2434 (Hz, system ABX, 6'-H), 2.94 and 2.99 (2s, 3H9N-CHS), 32 (dd i = 7.6 Hz and J ₂ = 7.4 Hz) and 332-3.43 (m)-[2H,-N-CH2-]/ 3.71 (d,lH,J=45 Hz,17 ^, β-H), 4.63 (ddd, IH, J16.15 = 10.2 Hz,.Ji6,i7-=45 Hz and Jι ₆ ,i5 3.9 Hz, 16'β-H), 6.50 (d, IH, J=24 Hz, 3'-OH), 6.60 (d, 1H,J=25 Hz, 4'-H), 6.66 (dd,lHJr=8.4 Hz and J ₂ =25 Hz, 2'H), 7.14 (d,lH,J=85 Hz, l'-H); IRVmaxGieat 3300, 1615. 1495 cm ¹ ; MS m/e 561,559 (M ⁺ , 40%, 100%), 523 (M ⁺ -HC1, 20%), 142 C ₂ H ₄ CON(CH3)C H9 ⁺ , 44%), 114 (C ₄ H ₉ (CH ₃ )CNO ⁺ , 37%); Exact mass calculated for C34H5 θ3N ³⁵ Cl 5593785, found 5593821.

- and -

-N-buty N-me yl-ll-(16 ^# α-cMoro-3\17 -Mihydroxy-estra-l ^/ 3 ^, 5 (10 ^/ )-trien-7 ^/ α- yl) undecanamide (*EM 139")

(25 mg, 55%) as a colorless oil; analytical sample was obtained by HPLC purification; 1H-NMR δ (CDCI ₃ , 400 MHz), 0.81 (s3H, 18'-CH ₃ ), 0.93 and 0.96 (2t, 3H,J=7.3 Hz, (CH ₂ ) ₃ CH ₃ ), 2.78 2H, T _6/6 =16.2 Hz and J ₆ , ₇ = 4.5 Hz, Δ5=24.34 HZ, system ABX, 6'-H), 2.94 and 2.99 (2s, 3H,N=CH ₃ ), 3.27 (dd, Jι-7.6 Hz and J ₂ -75 Hz)

and 331-3.45 (m, 2H, -N-CH ₂ ), 336 (dd, IH, Ji7,l7-OH=3.4 Hz and J ₁₇ , 6 = 5.9 Hz, 17'α-H), and 4.11 (ddd, IH, Jι ₆ ,i5 = 10.8 z J ₁₆ - ₁₇ = 5.9 z and Hz, 16'β-H), 6.56 (d, IH, J=19.7 Hz, 3'-OH), 6.61 (d, IH, J=25 Hz, 4'-H), 6.66 (dd, IH, Jι=8.4 Hz and J ₂ =2.6 Hz, 2'-H), 7.13 (d, IH, J=8.4 Hz, l'-H); IR v _max (neat) 3320, 1615. 1490 cm ¹ ; MS m/e 561,559 (M+, 38%, 100%), 523 (M ⁺ -HC1, 16%), 142 (C2H ₂ CON(CH3)C4H9 ⁺ , 80%), 114 (C4H ₉ (CH3)NCO ⁺ , 76%); exact mass calculated for 034H540SN35C15593785, found 5593825.

Scheme 9

10 : R={CH ₂ )ιoCO (CH ₃ )CH ₉

11 X=α, Br EM 105 X-Br, Rb=OH. Rtf=« EM 170 X = Br,R _tt =H.R _< -OH EM139 X«α,R _ti -H,R _< -OH EM 17lX-CI,R _b =H,R< _i =OH EM 156 X-=I.FI- _b =OH _I R _tf =H

N-n-butyl , N-methyl-1 l-(16'α-bromo-3'-acetoxy-l 7 ^, oxo-estra-l ' 3' 5'- (10 . trien-7'α-yl) undecanamide (11. X=Br)

To the above diacetate 10 (244 mg, 0.40 mmol) dissolved in 10 ml of acetic add 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 of acetic add, 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 50 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 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-lM16 ^, a-bromo-3'-acetoxy-17' -oxo-estra-l',3',5 ^, (10'), trien-7'-α-yl) undecanamide (11, X=Br) (201 mg, 78%), as colorless oil; ¹ H-NMR δ (CDC1 ₃ ), 0.94 (s, 3H,18'-CH3), 2-28 (s, 3H, 3'-OCOCH ₃ ), 2.82 app (d,lH,J=16.4 Hz, part of ABX system, 6'-H), Z90 and 2.96 (2s, 3H,H-CH ₃ ), 3.24 and 335 (2ta _PP ,2H, J=7.7 Hz, -N-CH ₂ -)d, 4.58 (t,lH,J=3.6 Hz, 16β-H), 6.82 (broad s,lH,4 ^, -H), 6.88 (dd,lH, J=8.0 Hz and J ₂ =4.0 Hz, 2'-H), 7.29 (d,lHJ=8 Hz, l'-H); MS m/e 644 ( M ⁺ ,7%), 565 (M ⁺ - Br, 77%), 522 (M ⁺ - B r - C O C H _{2 /} 55%), 142 (C ₂ H ₄ CON(CH ₃ )C ₄ H ₉ ⁺ .67%), 114 (C H ₉ (CH ₃ )NCO ⁺ _/ 66%), 88 (100%).

N-butyl. N-methyl-ll-(16'α-bromo-3'.17'-dihvdroxy-estra-l ^, 3 ^, .4 ^, (10 ^, )-trien-7'α- yl) undecanamide ("EM 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°C and a solution of 1.0 M of lithium aluminium hybride in ether (0.92 ml, 0.92 mmol) was added dropwise with rapid magnetic strirring. After 30 min, the reaction was quenched by the dropwise addition of a mixture of THF-ethyl acetate (1:1 v/v) and addified by 10% hydrochloric add. 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:

N-n-butyl. N-methyl-ll-(16 ^, α-bromo-3'.17'tt-dihydroxy-estra- V3'S (lθ')-trien- ^α-yl) undecanamide ("EM 171")

(63 mg, 21%) as colorless oil; Η-NMR δ (DCD1 ₃ , 400 MHz) 0.81 (s, 3H,18'-CH ₃ ), 0.93 and 0.96 (2t, 3H =73 Hz,N(CH ₂ ) ₃ CH ₃ ), 2.79 (2H,T _6/6 =16.6 Hz, j6,7= -7 Hz, = Δδ=24.34 Hz, system ABX,6'-H), 2.94 and 2.99 (2s H,N-CH ₃ ), 3.27 (dd,2HJι=7.7 Hz and J ₂ =7.5 Hz, -N CH ₂ -), 3.31-3.44 (m,2H,-N-CH ₂ -), 3.66 (dd,lH,Jι ₇ ,i7=1.4 Hz,

J _l7 , ₁ 6= 3 Hz, 17β-H), 4.68 6.60 (d, 1H =2.4 Hz, 4'-H), 6.65 (dd, 1H,J=8.5 Hz and J ₂ =25 Hz, 2'-H), 7.14 (d,lH,J=85 Hz, l'-H); IRVm _a x (neat) 3300, 1615, 1495 cm" ¹ ; MS m/e 605,603 (M ⁺ , 17%), 523 (M+-HBr, 81%), 142 (C ₂ H ₄ CON(CH ₃ )C ₄ H ₉ ⁺ , 100%), 114 (C4H ₉ (CH ₃ )NCO ⁺ , 97%); Exact mass calculated for C ₃₄ H5 ₄ 0 ₃ N ⁷⁹ Br 6033289, found 6033304.

- and -

N-n-butyl. N-methyl-ll-(16'α-bromo-3'. 17B-dihydroxy-estra-l'3 ^, .-5'(10')-trien- 7α-yl) undecanamide ("EM 105").

(170 mg, 50%) as a colorless oil; analytical sample was obtained by HPLC purification; Η-NMR δ (CDC1 ₃ , 400 MHz), 0.80 (s, 3H,18,-CH ₃ ), 0.93 and 0.96 (2t,3H,J=73 Hz,N(CH ₂ ) ₃ CH ₃ ), Z80 (2H,J ₆ ^=16.4,J6,7=4.6 Hz, Δδ=2434 Hz, system ABX, 6'-H), 2.94 and 2,99 (2s,3H,N-CH ₃ ), 3.27 (dd, 2H,Jι=7.7 Hz and J ₂ =75 Hz, -N-CH2-), 331-3.45 (m,2H,-N-CH2-), 4.02 Hz, 17α-H), 4.15 Hz, 16'β-H), 6.61 (d,lH,J=25 Hz, 4'-H), 6.66 (dd,lH,J=8.4 Hz and J2 25 Hz, 2'-H), 7.12 (d,lH,J=8.4 Hz, l'-H); IR v^ax (neat) 3320, 1610, 1490 cm" ¹ ; MS m/e 605, 603 (M+, 29%), 523 (M+-HBr, 100%), 142 (C2H4CON(CH ₃ )C ₄ H ₉ +, 70%), 114 ( H ₉ (CH ₃ )NCO ⁺ , 60%); Exact mass calculated for C ₃₄ H54θ ₃ N ⁷⁹ Br 6033289 found 6033289.

N-butyL N-methyl-ll-(16 ^, α-iodo-3M7'β-dihydroxy-estra-l ^, 3 ^, .-5'(10')-trien-7α- yl) undecanamide ("EM 156").

Under argon atmosphere, a mixture of 16α-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 h. 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 (1:1, v/v) gave a mixture of starting material and ϊodo compound (52:48) of which HPLC separation afforded N-butyl, N- memyl-ll,α6 ^, -α-iodo-3 ^, Λ7 ^, β-dihydroxy-cstra-l ^, ,3 ^, 5 ^, (10 ^, )-trien-7α-yl) undecanamide (ΕM 156") (21 mg, 36%) as colorless oil; Η-NMR δ (CDC1 ₃ , 00 MHz) 0.78 (s H,18'-CH ₃ ), 0.93 and 0.96 (2t3H =73 Hz, N(CH2) ₃ CH ₃ ), 2.79 (2H,J6,6=16.5 Hz,J ₆ ,7=4.4 Hz,Δδ, = 2434 Hz, system ABX, 6'-H), 2.94 and 2,99 (2s,3H,N-CH ₃ ), 3.27 (dd,2H ι-7.6 Hz and J ₂ -75 Hz, -N-CH ₂ ) 332-3.44 (m, 2H N-CH2), 4.09-4.17 (m, 2H, 16'βH and 17α-H), 6.60 (d,lH =2.4 Hz, 4'-H), 6.65 (dd,lH,J=8.4 Hz and J ₂ -2.4 Hz, 2'-H), 7.13 (d,lH,J=8.4 Hz, l'-H); IR v (neat) 3310, 1610, 1490 cm- ¹ ; MS m/e 651 (M+, 8%), 523 (M+-HI, 100%), 508 (M+-HI-CH ₃ ,38%) 142 (C ₂ H CON(C ^' H ₃ )C ₄ H ₉ ⁺ , 54%), 114 (C ₄ H ₉ (CH ₃ )NCO ⁺ , 49%); Exact mass calculated for C^^OsHI-HI 523.4028, found 523.4028.

EFFICACY OF AN ANTIESTROGEN SYNTHESIZED IN ACCORDANCE WITH EXAMPLE 9

Compound "EM 139" shown .., Sch le 9 a e is an estrogen activity inhibitor. "EM 139" has been tested botfr or efficacy in acting as an antiestrogen by blocking estrogen receptors without substantially activating those receptors (see Figure 3), and for efficacy in inhibiting 17β-hydroxysteroid dehydrogenase (see Hgure 4), an enzyme which catalyzes reactions involved in the synthesis of both androgens and estrogen (hereinafter referred to as "17β-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 subcutaneously in the appropriate groups in a solution of 0.9% (w/v) sodium chloride and 1% (w/v) gelatin at different concentrations of "EM 139" (as noted along the X axis of Hgure 3). 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 μg 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 art he rr s ± SEM of groups

of 9-10 mice. As may be seen from Figure 3, EM 139 was highly effective in reducing estradioi-induced uterine weight gain.

To test the effect of "EM 139" on the inhibition of sex steroid formation, its effect on the 17β-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 17β-hydroxystεroid dehydrogenase (Thomas et al., J. BioL Chem. 258: 11500-11504, 1983). 17β-hydroxysteroid dehydrogenase (17β-HSD) was purified to homogeneity from human placenta. A reaction vessel was prepared containing 1 μg 17β-HSD, 5 mM NAD, 20 μM 17β-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-HCl (50 mM), EDTA (2 mM), NaN ₃ (5 mM). The pH was 73. The reaction was allowed to proceed at 25°C for 15 min. Formation of NADH was measured at 340 nm. As shown by Figure 4, increasing concentrations of EM 139 significantly inhibited the reaction.

EXAMPLE 10

N-n-BUTYL, N-METHYL-ll-(3',17'β-DI-HYDROXY-1 α-ETHYNYL-ESTRA- WP-aw), 15'-TETRAEN-7'α-YL) UNDECANAMIDE ("EM 123") (SCHEME 10)

N-n-butyl. N-methyl-ll- 3'-benzoyloxy-1 α-ethylenedioxy est^a-l ^• 3 ^• 5'-(10 ^• )- ,trien-7 α-yl) undecanamide (12).

A πύxture of N-n-butyl, N-methyl-lK3 ^, -benzoyloxy-17 ^, -oxo estra-l^^OO')- trien-7'-α-yl) undecanamide (9a) (3.63 g), ethylene glycol (215 ml), p- toluenesulfonic add (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 (3x), 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 v/v) gave pure N-butyl, N-methyl-ll-(3'-benzoyloxy-17'-ethylenedioxy estra-l,3'5'(10'), trien-7'α-yl) undecanamide (3.58 g, 92%) as an oil, the structure of which was confirmed by spectroscopic means.

N-n-butyL N-methyl-ll-(3'-benzoyloxy-16'α-bromo-17'-ethylenedioxy-est ra-l' 3' 5'(10') . trien-7α-yl) undecanamide (13)

To the above ethylenedioxy amide 12 (370 mg, 055 mmol) in anhydrous tetrahydrofuran (10 ml) cooled at 0°C was added dropwise under argon, a solution of pyridinium bromide perbromide (406 mg, 136 mmol) in 7 ml of the same solvent After stirring during 15 h at 0°C, sodium iodide (300 mg) was added and the solution was stirred for 25 min. Afterwards, a solution of sodium thiosulfate (10%, v/v, 10 ml) and pyridine (05 ml) 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 IN hydrochloric add, water, saturated bicarbonate solution and water (3x), dried on magnesium sulfate and evaporated to dryness. The residue was chromatographed on silica-gd (50 g). Elution with a mixture of hexane-ethyl acetate (4:1 v/v) gave pure N-n-butyl, N-memyl-ll(3'-benzoyloxy-16'α-bromo- 17 ^* -ethylenedioxy-estra-l' ' 5' (10'), trien-7'α-yl) undecanamide (13) (313 mg, 76%) as colorless oil; IR vmax (neat), 1730, 1630, 1595 and 1255 cm" ¹ ; H NMR, 0.93 (3H, s, 18 ^* -CH ₃ ), 2 8 (2H, td, J = 75 and 2.6 Hz, -CH2CON-), 2.90 and 2.95 (3H, 2s, -N-CH ₃ ), 324 and 335 (2H, 2t, J = 73 Hz, -N-CH ₂ -), 3.85 and 435 (4H, m, -OCH2 Ch ₂ O), 4.56 (IH, m, H-C.16 ^* ), 6.91 (IH, d, J = 2.2 Hz, H-C.4') , 6.98 (IH, dd, J = 8.4 and 2.2 Hz, H-C2'), 732 (IH, d, J = 8.4 Hz, H-C.1'), 7.49 (2H, t _app J= 7.0 Hz H-C3" and H-C.5"), 7.63 (IH, t _app , J = 7.0 Hz H-C.4" and 8.17 (2H, d, J = 7.0

Hz, H-C2" and H-C6"), MS m/e, 671 (M+-Br,ll%), 114 ( H ₉ (CH ₃ ) NCO ⁺ , 13%), 105 (C6H ₅ ⁺ CO+, 100%), 86 ( H ₉ (CHs) N+, 10%), 77 (C6H ₅ ⁺ , 25%).

N-n-butyl. N-methyl-1 l-(3'-hydroxy-17'-oxo-estra-l' 3' 5'(10') . 15'-tetraen-7'α- yl) undecanamide "(EM 112)"

To a solution of the bromoketal (13) (517 mg, 0.69 mmol) in anhydrous dimethyl sulfoxide warmed at 73°C, under argon, was added potassium-t-butoxide (1.55 g, 13.8 mmol). The mixture was stirred for 5 h at this temperature and then cooled, poured in ice-water, addified with IN hydro-chloric add and extracted three times with ethyl acetate. The organic layers were washed with water (3x), dried on magnesium sulfate and evaporated to dryness. The residue was dissolved in acetone (30 ml), water (7 ml) and p-toluenesulfonic add (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 (3x), dried on magnesium sulfate and evaporated to dryness. The residue was purified by "flash chromatography" (100 g). Elution with a mixture of hexane-ethyl acetate (1:1 vv) gave the pure N-butyl, N-methyHl-(3 ^, -hydroxy-17 ^• -oxo-estra-l ^, ' 5' (10 ¹ ) ,15 ^* -tetτaen-7α-yl) undecanamide "EM 112" (178 mg, 49%) as colorless oil; IR v _max (neat), 3290,

1695, 1620 and 1600 cm" ¹ ; ^J H NMR, 0.92 and 0.95 (3H, 2t, J = 7.3 and 7.0 Hz, -N-(CH2) ₃ CH ₃ ), 1.11 (3H, s, 18"-CH ₃ ), 232 (2H, td, J = 25 and 7.0 Hz, H-C2), 2.94 and 2.99 (3H, 2s, N-CH ₃ ), 327 and 338 (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 = 2.6 Hz, H-C.4'), 6.71 (IH, dd, J = 8.4 and 2.6 Hz, H-C2% 7.13 (IH, d, J = 8.4 Hz, H-CD, 7.60 (IH, dd, J = 6.2 and 15 Hz, H-C.16') and 7.70 (IH, broad s, w _{1 2} =16 Hz, OH), MS m/e, 521 (M+, 53%), 507 (M+-CH ₂ ,9%), 506 (M+-CH _3/ 7%), 142 (C2H4 CON (CH ₃ ) C H9+ 25%), 114 OH^C-H^NCO- ¹ -' 60%) and 86 (C H ₉ (CH ₃ )N+, 22%, 44 (100%).

N-n-buty N-methyl-ll-(3'.17S-dihydroxy-17 ^, -α-ethynyl-estra-l ^r 3'5 ^, -(10').15'- tetraen-7'g-yl) undecanamide ("EM-123")

To hexanes (1 ml) cooled at 0°C, were added trimethylsilylacetylene (0.112 ml), n-butyllithium. 1.6 M in hexanes (025 ml), few drops of anhydrous THF and finally, a slowly addition of a solution of enone amide EM 112 (57 mg) in anhydrous THF (12 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 (3x). 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

mixture 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-gd (5 g). Elution with a mixture of hexanes: ethyl acetate (73 v/v) gave N-butyl, N-methyl-ll-(3',17 ^, β-dihydroxy- 17'α-ethynyl-estra-l',3'5 ^, -(10'),15'-tetraen-7O-yl)undecanamide ("EM-123") (34 mg, 63%); IR v _max (neat), 3290, 2150, 1620 and 1600 cm" ¹ ; lH NMR, 0.92 and 0.95

(3H, 2t, J = 73 and 7.0 Hz, N- CH _∑ bCHs), 0.95 (3H, s, lδ'-CHs), 232 (2H, td, J = 7.0 and 2. Hz, -CH2CON-), 2.66 (IH, s, -CH), 2.93 and 2.98 (3H, 2s, N-CHb), 3.27 and 3.38 (2H, t, J = 7.0 Hz, -N-CH2-), 5.78 (IH, dd, j = 5.9 and 33 Hz, H-C.15% 6.05 (IH, dd, j = 5.9 and 15 Hz, H-C.16'), 6.62 (IH, d, J = 25 Hz, H-C.4'), 6.67 (IH, dd, J = 8.4 and 2.6 Hz, H-C2') and 7.13 (IH, d, J = 8.4 Hz, H-Cl') ppm; MS m/e 547 (M+, 12%), 142 (C2H CON(CH ₃ )C4H9+, 21%) 114 (G|H ₉ (CH ₃ )NCO ⁺ , 50%), 88 (100%) and 86 (C4H ₉ (CH ₃ )N+, 34%).

EXAMPLE 11

16β-CYCLOPROFYL DERIVATIVES

(Scheme 11)

N-n-butyL N-methyl-ll-(17'-oxo-3'-hydroxy-15'β.l6'β-metnylene-estra l'3'5'(10')-trien-7'tt-yD undecanamide (14)

A solution of the phenol-enone EM-112 (101 mg; 0.19 mmol) dissolved in anhydrous pyridine (15 ml) and acetic anhydride (10 ml) 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 add, 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 g). Elution with a mixture of hexane-ethyl acetate (73 v/v) gave the N-butyl, N-mefhyl-ll-(17'-oxo-3'- acetoxy-estra-l'^ ^, 5'(10'),15'-tetraen-7'ct-yl) undecanamide.

To this and palladium (ID acetate (11 mg) in ether (25 ml) an ethereal diazomethane solution (prepared from 1 g of diazald) was added dropwise at 0°C with continuous stirring during 10 min. After evaporation, the residue was dissolved in methanol (50 ml) and cooled to 0°C. 2N sodium, hydroxyde solution (1ml) was added and after 75 min. of stirring the mixture was neutralized with IN hydrochloric add, extracted three times with ether. The organic layers were washed with brine, dried on magnesium sulfate and

evaporated to dryness. The residue was purified by HPLC to give N-butyl, N- methyl-ll-(17 ^, -oxo-3 ^, -hydroxy-15 ^, β, 16 ^, β-methylene-estra-r3 ^, 5'(10 ^, )-trien-7 ^, α- yl) undecanamide (14) (79 mg, 76%) as a colorless oil. IR v _max (neat) 3260, 1705,

1610 and 1570 cm" ¹ ; ¹ H NMR (400 MHz) 0.93 and 0.96 (3H, 2t, J = 7.3 Hz, NfCH _∑ bCHs), 0.99 (3H, s, 18'-CH ₃ ), 1.98 (IH, td, J= 83 and 3.% Hz, H-C16'), 2.80 (IH, d, J = 16.6 Hz, Hβ-C.6% 2.94 and 2.98 (3H, 2s, N-CH ₃ ), 3.27 (IH, dd, J = 758 and 6.66 Hz) and 338 (IH, m) (both are -N-CHr), 6.64 (IH, d, J = 2.6 Hz, H-C4'), 6.66 (IH, dd, J = 82 and 2.6 Hz, H-C.3") and 7.10 (IH, d, J = 8.2 Hz, H-C.1') ppm; MS m/e 535 (M+, 74%), 522 (M+-CH2, 49%), 129 (C4H ₉ (CH ₃ )NCOCH ₃ +, 37%), 114 ( H ₉ (CH ₃ )NCO+, 67% and 88 (100%).

N-n-butyl. N-methyl-11 -(3'. 17 ^, β-dihydroxy-15'S.16'B-methylene-estra- l'3'5'(10 ^, )-trien-7'α-yl) undecanamide (EM-136)

To the cydopropylketone 14 (10 mg, 18.7 μmol) 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

silica-gel (5 g). Elution with a mixture of hexanes: ethyl acetate (5:5 v/v) gave N-b tyl,N-methyl-ll-(3 ^, ,17 ^, β-dihydroxy-15'β,1 ^, -cydopropyl-est^a-l ^, 3'5 ^, (10 ^, )- trien-7'α-yl) undecanamide ("EM-136"), as a colorless oil, IR v _max (neat)

3300,1615, 1580 and 1450 cπr ¹ , *H NMR (400 MHz), 031 (IH, dd, J = 14.0 and 7.8 Hz, H-Cl") 033 (3H, s, lβ'-CHs), 0.93 and 0.% (3H, 2t, J = 73 Hz, N(CH2) ₃ CH3), 2.77 (IH, d, J = 17.1 Hz, Hβ-C.6'), Z94 and Z98 (3H, 2s, N-CH ₃ ), 3.27 (IH, dd, J = 7.7 and 73 Hz) and 339 (IH, m) (both are -N-CH2-), 4.09 (IH, broad s, w = 10 Hz, H-C17 , 6.64 (2H, m, H-C4' and H-C2") and 7.11 (IH, d, J = 83 Hz, H-C.l') ppm; MS m/e 537 (M+, 18%), 519 (M+-H ₂ 0, 56%), 504 (M+-H ₂ 0-CH ₃ , 100%), 142 (C ₂ H4CON(CH ₃ )C4H9 ⁺ , 70%), 114 (C4H ₉ (CH3)NCO ⁺ 50%) and 86 (C ₄ H9(CH ₃ )N ⁺ 33%).

N-n-butyl. N-methyl-ll-(3M7 ^, 6-dihydroxy-17'α-ethynyl-15'β.l6'β-methylene- estra-l ^, 3'5'(10 ^, )-trien-7 ^, α-yl) undecanamide ("EM-138")

To hexanes (500 μl) cooled at 0°C, were added trimethylsilylacetylene (54.6 μl), 1.6 M n-butyl lithium in hexanes (120.4 μl), few drops of anhydrous THF and finally, a slowly addition of a solution of the cydopropyl ketone 14 (25.8 mg) in anhydrous THF (350 μl). The mixture was stirred for 75 min at 0°C. 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

brine, dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure. To the residue dissolved in methanol (900 μl), a 5N potassium hydroxyde solution (70 μl) was added and the mixture refluxed for 30 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 g). Elution with a mixture of hexanes: ethyl acetate (5:5 v/v) gave N-butyl, N-methyl-ll-(3',17β-dihydroxy- 17 ^, α-ethynyl-15 ^, β,16'β-cydopropyl-estra-l'3 ^, 5'(10 ^, )-trien-7 ^, α-yl) undecanamide ("EM 138") (12 mg, 44%) as a colorless oil; IR v _max (neat) 3590, 3300, 1620, 1600 and 1450 cm" ¹ ; *H NMR (400 MHz), 039 (IH, ddd, J = 14.6 and 7.9 Hz, H-C.1'), 0.93 and 0.% (3H, 2t, J = 7.4 and 73 Hz, -N(CH ₂ ) ₃ -CH ), 0.96 (3H, s, 18'-CH ₃ ), 2.70 (IH, s, -C CH), 2.77 (IH, d, J = 165 Hz, Hβ-C.6'), 2.94 and 2.98 (3H, s, N-CHs), 3.27 (IH, dd, J = 7.7 and 7.6 Hz) and 338 (IH, m) (both are N-CH2-), 6.42 (IH, m, OH), 6.65 (2H, m, H-C4' and H-C2') and 7.12 (IH, d, J = 83 Hz, H-C.1') ppm; MS m/e 561 (M+, 15%), 142 (C2H4CON(CH ₃ )C ₄ H ₉ +, 66%), 114 ( H ₉ (CH ₃ )CO ⁺ , 53%), 88 (100%) and 86 ( H ₉ (CH ₃ )N+, 35%).

Scheme 11

9a 12

13

EM 123 EM 112

EM 136 EM 138

EXAMPLE 12 17α-ALKYNYLAMIDE ESTRADIOLS

GENERAL PROCEDURE FOR ESTER FORMATION (SCHEMA 12)

In anhydrous conditions under argon atmosphere, bromo add (17 mmol) was dissolved in dry CH2CI2 (30 ml), oxalyl diloride (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 CH2CI2 (10 ml) and added at 0°C to a solutin of 3-methyl 3-oxetanemetha ol (17 mmol), CH2CI2, (7 ml) and pyridine (1.4 ml). The reaction was kept at this temperature for 4-8 h. Thereafter, mbcture was diluted with CH2CI2, washed with NaHC0 ₃ (10%, v/w) and organic phase was dried over MgSθ4« 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-methyl-3-hydroxymethyloxetane (15).

Light yellow liquid (91% yield); IR v (neat) 2930, 2860, 1725 1450, 1370, 1160 cm" ¹ ; NMR-60 δ (CDC1 ₃ ) 131 (s, 3H), 1.1-2.1 (m, 6H), 236 (t, J = 6.0 Hz, 2H), 3.36 (t, J = 6 Hz, 2H), 4.13 (s, 2H), 4.41 (AB system ΔV -= 83, J = 6 Hz, 4H).

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

Colorless liquid (86% yield); IR v (neat) 2920, 2840, 1725, 1450, 1370, 1150 cm" ¹ ; NMR-60 δ (CDC13) 131 (s, 11H), 12-22 (m, 4H), 2.40 (t, J = 6.0 Hz, 2H), 3.45 (t, J = 6.0 Hz, 2H), 420 (s, 2H), 4.20 (s, 2H), 4.48 (AB system ΔV = 8.2, J = 6.0 Hz, 4H).

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

Colorless liquid (85% yield); NMR-60 δ (CDC1 ₃ ) 133 (s, 15H), 1.0-2.0 (m, 4H), 230 (t, J = 6.0 Hz, 2H), 335 (t, J = 6.0 Hz, 2H), 4.12 (s, 2H), 4.40 (AB system ΔV - 8.2, J = 6.0Hz, 4H).

12-bromo dodecanoate ester of 3-methyl-3-hydroxymethyl oxetane (18).

Colorless liquid (86% yield): IR v (neat) 2910, 2840, 1720, 1450, 1370, 1155 cm" ¹ ; NMR-60 δ (CDC1 ₃ ) 130 (s, 17H), 1.1-2.0 (m, 4H), 2.30 (t, J = 6.0 Hz, 2H), 3.33 (t, J - 6.0 Hz, 2H), 4.11 (s, 2H), 4.40 (AB system Δ v = 8.0, J - 6.0 Hz, 4H).

GENERAL PROCEDURE FOR OR HO ESTER FORMATION (SCHEME 12)

To a solution of bromo ester (3.4-142 mmol) in dry CH2CI2 (10-40 ml) at 0°C was added with stirring distilled boron trifluoride etherate (0.85-335 mmol). After 4 h at 0°C, reaction mixture was quenched by the addition of triethylamine (3.4-142 mol), diluted with diethylether and filtered to remove the amine-BF ₃ complex. The filtrate was evaporated and residue was purified by chromatography (hexane-ethylacetate-triethylamine / 80:20:1, v/v/v) to give bromo ortho ester.

l-(5'-bromo pentanyl)-4-methyl-2.6,7-trioxabicydo f222l octane (19).

Colorless oil (68% yield); IRv (neat) 2940, 2915, 2855, 1450, 1390, 1050, 980 cm" ¹ ; NMR-60 δ (CDC1 ₃ ) 0.79 (s, 3H), 1.2-2.0 (m, 8H), 335 (t, J = 6.0 Hz, 2H), 3.87 (s, 6H); MS m/e (rel. intensity) 280 (M+, 02), 278 (M+, 0.2), 250 (8.1), 248 (83), 197 (7.2), 195 (7.7), 179 (58), 177 (61), 72 (54), 69 (100).

l-(8'-bromo octanyl)-4-methyl-2.6.7-trioxabicydo [2.2.21 octane (20).

Amorphous white solid (69% yield); IR v (KBr) 2940, 2900, 2840, 1450, 1390, 1045, 985, 950 cm- ¹ ; NMR-60 δ (CDC1 ₃ ) 0.80 (s, 3H), 133 (s, 8H), 1.0-2.1 (m, 6H), 3.40 (t, J = 6.0 Hz, 2H), 3.93 (s, 6H); MS m/e (rel. intensity) 323 (M ⁺ , 2.1), 321 (M ⁺ , Z0), 292 (4.4), 290 (5.1), 239 (8.6), 237 (7.1), 221 (43), 219 (33), 69 (71), 55 (100).

l-(10'-bromo decanyl)-4-methyl-2.6.7-trioxabicvdo f2.2.2l octane (21).

White solid (74% yidd); m.p. 51-53°C; IR v (KBr) 2940, 2900, 2850, 2830, 1455, 1390, 1055, 985, 955 cm" ¹ ; NMR-60 δ (CDC1 ₃ ) 0.80 (s, 3H), 127 (s, 12H) 1.1-2.1 (m, 6H), 339 (t J = 6.0 Hz, 2H), 3.87 (s, 6H); MS m/e (rel. intensity) 350 (M+, 1.2), 348 (M ⁺ , 1.1), 321 (3.0), 319 (7.6), 269 (73), 248 (97), 144 (37) 55 (100).

l-(ll'bromo undecananyl)-4-methyl-2.6.7.trioxabicydo T2221 octane (22).

White solid (76%, yield); m.p. 473, 483°C; IRv (KBr) 2900, 2835, 1460, 1045, 975 cm" ¹ ; NMR-60 δ (CDC1 ₃ ) 0.79 (s, 3H), 125 (s, 14H), 1.1-2.1 (m, 6H), 337 (t, J = 6.0 Hz, 2H), 335 (s, 6H); MS m/e (rd. intensity) 364 (M+, 33), 362 (M+, 3.4), 334 (13), 332 (13), 283 (15), 263 (85), 261 (97), 144 (51), 55 (100).

4. Preparation of 17α-alkynylamide estradiols (scheme 13)

GENERAL PROCECURE FOR COUPLING REACTION (Scheme 13)

In a flame-dried flask under argon atmosphere, 3,17β-bis tetrahydropyranyl ethynylestradiol 23 (13 mmol) synthesized from commerdal 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 ^e 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 MgSθ ₄ and the solvent was removed under reduced pressure. The crude produrt was purified by chromatography (hexane-ethylacetate-triethylamine / 96:4:1 to 5030:1, v/v/v) to give coupling product 24-27, unreacted steroid 23 (61, 62, 57%) and small quantity of undeterminated product.

l-f3M β-bis r(tetrahydrc _^ 2Η-pyran-2"yl)oxyl estτa-l ^, 3'3'(10')-trien-17a-yn-7- '-methyl-T.O'y'-rrioxapicyclo f2'.2'2'1 octan-l'-ylH-heptyne (24).

Colorless oil (15% yield); IR v (neat) 2920, 2855, 2230, w, 1600, 485 cm" ¹ ; NMR-60 δ (CDC1 ₃ ) 0.75 (s, 3H), 0.88 (s, 3 H), 2.80 (n, 2H), 3.2-4.1 (m, 4H), 3.80 (s, 6H), 4.9-5.3

(m, IH), 534 (s, IH), 6.75 (m, 2H), 7.19 (d, J = 8.0 Hz, IH); MS m/E ^' (rel. intensity) 579 (M+-DHP, 4.0), 564 (1.1), 494 (12), 477 (12), 374 (13), 85 (1000.

(4 ^, -methyl-2'.6 ^, .7 ^, -trioxabicydo r2'2'2'l octan-l'-yD- -decyne (25).

Colorless oil (15% yield); IR v (neat) 2915, 2850, 2210 w, 1600, 1485 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.79 (s, 3H), 0.90 (s, 3H), Z24 (t, J = 6.6 Hz, 2H), 2.83 (m, 2H), 335 (m, 2H), 3.89 (s, 6H), 3.95 (m, 2H), 4.98 and 5.19 (2s, IH), 539 (s, IH), 6.78 (d, J = 2.6 Hz, IH), 6.84 (dd, Ji = 26 Hz and J ₂ = 8.4 Hz, IH), 7.22 (d, J = 8.4 hZ, IH); MS m/e (rd. intensity) 620 (M+-DHP, 43), 535 (13), 518 (8.9), 85 (100).

l-f3'.17B-bis f(tetrahvdro-2'Η-pyran-2 l)oxy1 estra-r3'3 ^, .(10')-trien-17α-yiμi2- (4'-methyl-2'.6',7'-trioxabicydo r2'2'2'1 octan-l'-yl)-l-dodecvne (26).

Colorless visquous oil (42% yidd); IR v (neat) 2920, 2850, 2210 vw, 1600, 1485 cm ¹ ; NMR-200 δ (CDC1 ₃ ) 0.79 (s, 3H), 0.90 (s, 3H), 225 (t, J = 6.6 Hz, 2H), 2.83 (m, 2H), 3.55 (m, 2H), 3.89 (s, 6H), 3.95 (m, 2H), 5.0 and 5.2 (2s, IH), 539 (s, IH), 6.78 (d, J = 2.6 Hz, IH), 6.84 (dd, Ji = 2.6 and J ₂ = 8.4 Hz, IH), 7.21 (d, I = 8.4 Hz, IH); MS m/e (rel. intensity) 649 (M ⁺ -DHP, 6.1), 634 (0.7), 564 (22), 547 (16), 85 (100).

l-(3'.1 β-bisf(tetrahvdro-2"H-pyran-2''yl)oxylestra-l'3'5'(10')-tτ ien-l-17'α-yl)-13- (4'methyl-2',6'.7-trioxabicvdor2'.2'2 ^, 1octan-l'-yl)-l-tride cyne (27)

Colorless visquous oU (35% yidd); IR v (neat) 2915, 2850, 2290 vw, 1600, 1490 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.80 (s, 3H), 0.90 (03H), 2.25 (t, J = 6.6 Hz, 2H), 2.83 (m, 2H), 333 (m, 2H), 3.89 (s, 6H), 3.95 (m, 2H), 5.0 and 5.2 (2s, IH), 539 (s, IH), 6.78 (d, J = 2.2 Hz, IH), 6.84 (dd, Jι= Z6 and J ₂ = 8.4 Hz, IH), 7.21 (d, J = 8.4 Hz, IH).

GENERAL PROCEDURE FOR ORTHO ESTER AND DI-THP HYDROLYSIS

The product with ortho ester and di-THP group (0.22-0.63 mmol) was dissolved in MeOH (80-120 ml) and p-toluenesulfonic add (0.17-023 mmol) was added. The solution was stirred at room temperature for 2-3 h. Then, water was added, MeOH was removed under reduced pressure and residue was extracted with etiiylacetate. After .evaporation of solvent, the crude product was purified by column chromatography (hexane-ethylacetate / 5;5, v/v) to give ester compound with free hydroxyl group.

8-(3M76-dihydroxy estra-l ^, 3'3'(10')-trien-17'α-yl)-7-octynoate ester of 2'2'- dihydroxymethyl propanol (28).

Colorless visquous oil (70% yield); IR v (film) 3340, 2910, 2850, 1710, 1600, 1485 cm" ¹ ; NMR-200 δ (CDQ ₃ ) 0.83 (s, 3H), 0.86 (s, 3H), 227 (t, J = 6.4 Hz, 2H), 238 (t, J = 7.1 Hz, 2H), 2.81 (m, 2H), 334 (s broad, 4H), 4.17 (s, 2H), 4.87 (s, IH), 636 (d, J = 2.6 Hz, IH), 6.63 (dd, Ji = 2.6 Hz and J ₂ = 8.4 Hz, IH), 7.17 (d, J = 8.4 Hz, IH); MS m/e (rel. intensity) 512 (M+, 14), 494 (97), 479 (17), 466 (11), 270 (48), 159 (100).

ll-(3'.17B-dihyroxy estra-l'3'3'(10')-trien-17 ^, α-yl)-10-undecynoate ester of 2'2'- dihydroxymethyl propanol (29).

Colorless visquous oil (61% yidd); IR v (neat) 3360, 2910, 2840, 2210 vw, 1710, 1600, 1485 cm- ¹ ; NMR-200 (CDC1 ₃ ) δ 034 (s, 3H), 0.86 (s, 3H), 2.24 (t, J = 7.0 Hz, 4H), 2.79 (m, 2H), 334 (s broad, 2H), 336 (s broad, 4H), 4.13 (s, 2H), 637 (s _app , IH), 6.63 (dd, Ji = 2.6 Hz and fc = 8.4 Hz, IH), 7.14 (d, J = 8.4 Hz, IH) MS m/e (rel. intensity) 554 (M+, 5.0), 536 (57), 520 (10), 507 (7.6), 435 (14), 419 (20), 270 (39), 160 (85), 133 (100).

13-(3'. 17'β-dihvdroxy estra-l'3'3'(10')-trien-17α-yl)-12-tridecynoate ester of 2'2'-dihydroxymethyl propanol (30).

Colorless visquous oil (78% yield); IR v (film) 3360, 2915, 2840, 1710, 1600, 1490 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.83 (s, 6H), 225 (m, 4H), 2.78 (m, 2H), 3.53 (s broad, 4H), 4.09 (s, 2H), 6.6 (m, 2H), 7.10 (d, J = 8.0 Hz, IH); MS m/e (rel. intensity) 582 (M+, 1.0), 563 (38), 548 (5.7), 535 (33), 463 (5.7), 446 (13), 270 (44), 160 (57), 133 (58), 55 (100).

14-(3M7 ^, B-dihvdroxy estra-l'3'3'(10')-trien-17 ^t α-yl)-13-tetradecvnoate ester of 2'2'-dihydroxymethyl propanol (31).

Colorless visquous oil (83%, yield); IR v (film) 3360, 2910, 2840, 2220 vw, 1710, 1605, 1490 cm- ¹ ; NMR-200 δ (CDd ₃ ) 035 (s, 3H), 0.87 (s, 3H), 225 (t, J = 6.6 Hz, 2H), 233 (t, J = 7.1 Hz, 2H), 230 (m, 2H), 23 (m, 2H), 338 (s broad, 4H), 420 (s, 2H), 572 (s, IH), 636 (d, J = 2.6 Hz, IH), 6.62 (dd, ft =2.6 Hz and J ₂ = 8.4 Hz, IH), 7.15 (d,J=8.8 Hz,lH).

GENERAL PROCEDURE FOR HYDROLYSIS OF ESTER FOLLOWING BY AMIDE FORMATION

At a solution of ester (0.14-0.49 mmol) in MeOH (12-50 ml) was added aqueous solution of KOH 10% w/v (6-25 ml) and mixture was refluxed under argon atmosphere for 24 h. Thereafter, water was added and MeOH was evaporated under reduced pressure. The resulting solution was addified with HCl and extracted with ethylacetate. Organic phase was washed with water, brine and dried over MgSO Without purification, the crude carboxylic add (IR add band at 1700 and 2400-3600 cm" ¹ ) was dissolved in dry CH2CI2 (20-70 ml) and tributylamine (0.58-2.04 mmol). The mixture was cooled at -10 ^* C, isobutyl chloroformate (0.68-241 mmol) was added and allowed to react 30 min. At this time, N-methylbutylamine in excess (42-16.0 mmol) was added and the cooling bath was removed. After 2 h, CH2CI2 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/ 73, v/v).

N-buty N-methyl-8-r3'-(i-butyloxy carbonyloxy)-17B-hydroxy estra-l'3'3 ^, (10')- trien-17'α-yll-7-octynamide (32).

Colorless oil (79% yidd); IR v (neat) 3380, 2920, 2850, 1745, 1620 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.87 (s, 3H), 0.91 and 0.94 (2t, J = 73 Hz, 3H), 1.00 (d, J = 6.6 Hz, 6H), 235 (m, 2H), 289 and 291 (2s, 3H), 322 and 333 (2t, J = 73 Hz, 2H), 4.02 (d, J = 7.0 Hz, 2H), 638 (d, J = 26 Hz, IH), 6.93 (dd, = 26 Hz and J ₂ = 8.4 Hz, IH), 729 (d, J = 8.4 Hz, IH); MS m/e (rd. intensity) 579 (M+, 12), 561 (26), 546 (11), 461 (6.7), 447 (3.7), 270 (84), 57 (100). EMS M+ calculated for CsβHssOsN: 5793923; found: 5793970.

N-butyl, N-methyl-ll-f3'-(i-butyloxy carbonyloxy)-17'B-hydroxy estra- l^' lO^-trien-lT-g-yn-lO-undecvnamide OS).

Colorless oil (67% yield); IR v (neat) 3370, 2910, 2840, 1745, 1620 cm" ¹ ; NMR-200 δ (CDα ₃ ) 037 (s, 3H), 0.92 and 0.95 (2t, T = 6.6 Hz, 3H), 1.00 (d, J = 7.0 Hz, 6H), 286 (m, 2H), 290 and 294 (2s, 3H), 324 and 335 (2t, J = 73 Hz, 2H), 4.03 (d, J = 6.6 Hz, 2H), 6.88 (d, J = 26 Hz, IH), 6.93 (dd, Ji = 26 Hz and J ₂ = 83 Hz, IH), 730 (d, J = 8.1 Hz, IH); MS m/e (rel. intensity) 621 (M ⁺ , 21), 606 (24), 602 (6.2), 212 (43), 159 (69), 142 (68), 114 (100).

N-butyl. N-methyl-13-f3 ^' (i-butyloxy carbonyloxy)-17β-hydroxy estra-l',3',5'(10')- trien-17g-ylH2-tridecynamide (34).

Colorless oil (89% yield); IR v (neat) 3370, 2920, 2840, 1745, 1620 cm" ¹ ; NMR-200 δ (CDQ ₃ ) 0.87 (s, 3H), 0.92 and 0.95 (2t, J = 7.0 Hz, 3H), 1.00 (d, J = 7.0 Hz, 6H), 286 (m, 2H), 290 and 296 (2s, 3H), 325 and 335 (2t, J = 7.4 Hz, 2H), 4.02 (d, J = 6.6 Hz, 2H), 6.88 (d, J = 22 Hz, IH), 6.93 (dd, Ji = 26 Hz and J ₂ = 8.4 Hz, IH), 730 (d, J = 8.8 Hz, IH); MS m/e (reL intensity) 649 (M+, 20), 633 (15), 631 (18), 616 (8.2), 531 (15), 516 (5.6), 270 (85), 57 (100); EMS M+ calculated for iH∞OsN: 649.4706; found: 649.4643.

N-butyl. N-methyl-14-r3'(i-butyloxy carbonyloxy)-17B-hvdroxy estra-l'3'5'(10')- trien-17α-yl1-13-tetradecynamide (35).

Colorless oil (83% yield); IR v (neat) 3380, 2910, 2840, 1750, 1625 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.87 (s, 3H), 0.92 and 0.95 (2t, J = 7.0 Hz, 3H), 1.00 (d, J = 6.6 Hz, 6H), 2.85 (m, 2H), 2.91 and 2.96 (2s, 3H), 3.25 and 336 (2t, J = 7.4 Hz, 2H), 4.03 (d, J = 6.6 Hz, 2H), 6.88 (d, J = 26 Hz, IH), 6.93 (dd, Ji = 29 Hz and J ₂ = 8.4 Hz, IH), 730 (d, J = 8.8 Hz, lH).

HYDROLYSIS OF CARBONATE

Hydrolysis of carbonate compounds 32-35 was performed as follows: carbonate derivatives were dissolved in methanol (10 ml). K ₂ C0 ₃ (1%; 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 addified with HCl (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 6333, v/v).

N-butyl. N-methyl-8-r3M7B-dihydroxy estra-l ^, 3 ^, 3'(10')-trien-17'g-yll-7- octynamide ("EM 157").

Purified by column chromatography (hexane-ethyl acetate/4:6, v/v). Amorphous white solid (88% yield); IR v (film) 3280, 2910, 2840, 1610 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.87 (s, 3H), 0.91 and 0.94 (2t, J = 7.0 Hz, 3H), 280 (m, 2H), 2.90 and 292 (2s, 3H), 3.22 and 334 (2t, J = 7.3 Hz, 3H), 5.22 (s, IH), 637 (d, J = 29 Hz, IH), 6.64 (dd, = 26 Hz and J = 8.4 Hz, IH), 7.16 (d, J = 8.4 Hz, IH); MS m/e (rel. intensity) 479 (M ⁺ , 11), 462 (18), 460 (38), 446 (18), 270 (30), 114 (56), 88 (67), 44 (100); EMS M+ calculated for iH-isOsN: 479.3399; found 4793369.

N-butyl. N-methyl-1 l-r3M7'B-dihvdroxy estra-l ^, 3'3 ^, (10 ^, )-rrien-17'α-yl)-10- undecynamide ("EM 183").

Purified by column chromatography (hexane-ethylacetate/4:6, v/v). Amorphous white solid (83% yidd); IR v (KBr) 3300, 2910, 2840, 1610 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.87 (s, 3H), 0.93 and 0.95 (2t, J = 7.0 Hz, 3H), 280 (m, 2H), 291 and 294 (2s, 3H), 3.23 and 335 (2t, J = 73 Hz, 2H), 530 (s, IH), 637 (d, J = 26 Hz, IH), 6.64 (dd, Ji = 26 Hz and J ₂ = 8.4 Hz, IH), 7.16 (d, J = 8.1 Hz, IH); MS m/e (rd. intensity) 521 (M+, 4.4), 505 (10), 502 (26), 489 (7.7), 487 (8.7), 270 (20), 114 (55), 88 (42), 44 (100).

N-butyl. N-methyl-13-f3M7B-dihydroxy estra-l'3'3 ^, (10')-trien-17g-yn-12- tridecynamide ("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 δ (CDC1 ₃ ) 0.88 (s, 3H), 0.93 and 0.95 (2t, J = 7.0 Hz, 3H), 280 (m, 2H), 2.93 and 2.97 (2s, 3H), 325 and 338 (2t, J = 73 Hz, 2H), 6.61 (d, J = 2.6 Hz, IH), 6.69 (dd, = 2.6 Hz and J ₂ = 8.6 Hz, IH), 6.87 (s, IH), 7.14 (d, J = 8.1 Hz, IH); MS m/e (rel. intensity) 549 ,(M+, 8.7), 532 (17), 530 (23), 516 (12), 270 (30), 114 (35), 88 (45), 44 (100); EMS M ⁺ calculated for C^HssOs : 549.4182, found: 549.4271.

N-butyl, N-methyl-14-r3'.17'β-dihydroxy estra-l ^, 3'3 ^, (10')-trien-17 ^, α-yll-13- tetradecvnamide ("EM 196").

Purified by column chromatography (hexane-ethyl acetate/ 6:4, v/v). Amorphous white solid (93% yield); IR v (film) 3280, 2915, 2840, 1615 cm" ¹ ; NMR-200 δ (CDC1 ₃ ) 0.88 (s, 3H), 0.94 and 0.95 (2t, J = 7.0 Hz, 3H), 280 (m, 2H), 295 and 298 (2s, 3H), 326 and 339 (2t, J = 73 Hz, 2H), 6.61 (d, J = 22 Hz, IH), 6.70 (dd, Ji = 2.6 Hz and fe = 8.4 Hz, IH), 7.13 (m, 2H: aromatic and phenolic hydrogen).

Scheme 12

Br(CH2)nCOOH >- C-CH ₃

15-18 19-22

(n-5.8.10.11) (n-5. ^β .10.m

Scheme 13

EIHINYL- ESTRADKDL

EΞM 157 n-5 EM 183 n-8 EM 163 n-10 EM196 n-11

EXAMPLE 13 Scheme 14

N-n butyl, N-methyl-3(3 ^' -17β-dihydroxy-ll'β-methoxy estra l'^'dO -trien 7α-yl) undecanamide ("EM 111") and its 17α-ethynyl derivatives ("EM 121").

gBrfC+ynOTHP

Cud

142-

46 47

- 144 -

EXAMPLE 14 Scheme 15

ll _β -chloromethyl derivatives

N-n-butyl, N-methyl, (llβ-chloromethyl-3',17β-dihydroxy-estra-l',3',5'(10 trien-7α-yl) undecanamide (56) and its 17α-ethynyl derivat ⁱ ve (58)

51 52

53

-146-

EXAMPLE 15 Scheme 16

Compounds with aliphatic side-chain in 17β-position

N-n-butyl, n-methyl, ll-07β-hydroxy-3*-methoxy estea-l' 3',5 ^" (10 -trien- 17α'α-yl) undecanamide ("EM 103").

63 RJiCH, 64

65 66

EM 103

EXAMPLE 16 Scheme 17

17α-cydopropyl derivatives

N-n-butyl, N-methyl-(17α-cydopropyl-3',17β-dihydroxy 7α-yl) undecanamide (68) and its 17α-chlorocydopropyl and 17α- fluorocydopropyl derivative (69)

EXAMPLE 17

N-n-butyl, N-methyl-(17α-cyclopropyl-3',17β-dihydroxy ll'β-methoxy estra undecanamide (70 X = H) and its 17α-chlorocylopropyl and 17α-fluorocydopropyl derivative (70, X = F, Cl)

Same example as Example 16 with compound 49 as starting material.

EXAMPLE 18 Scheme 18

17α-cyanovinyl derivatives

N-n-butyl, N-methyl-ll-(17'α-cydopropyl-3 ^* ,17β-dihydroxy ll'β-methoxy estra undecanamide (73)

B = 1) Meϋ, CO ₂ 2) NH3

3) NaBH ₄ -pyr

4) H ₂ , Lindlarcat.

EXAMPLE 19 Scheme 19

Compounds with aliphatic side chain in 15α-position

N-n-butyl, N-memyl-ll-(3 ^* ,17β-dihydroxy 17α-ethynyl estra l' ',5 ^, ,(10)-trien- 15'α-yl) undecanamide ("EM 108")

CH3COCH3 pTsOH

152

1)CICO ₂ R 2) CK ₃ NHC ₄ H ₉

EXAMPLE 20 Scheme 20

17α-thioethyl derivatives

N-n-butyl, N-methyl-1 l-(3',17β-dihydroxy 17α-thioethyl estra V3'P,(lO)-t ^ή en- 7α-yl) undecanamide (82) and its ethyl disulfite derivative (83)

EXAMPLE21 Sch me 2.1

17α-thiopropyl derivatives

N-n-butyl, N-methyHl-(3 ^' ,17'β-dihydroxy 17α-thiopropyl (10')- trien-7-α-yl) undecanamide (84) and its ethyl disulfite derivative (86)

Etθ2θ-N=N-CO ₂ Et PPh ₃

EXAMPLE 22 Scheme 22

llβ-ethyl derivatives

N-n-butyl, N-methyl-ll-(3 ^, ,17'β-dihydroxy 17α-ethynyl-llβ-ethyl estra l^^'dO rien^α-yl) undecanamide (97)

-156-

EXAMPLE 23 Scheme 23

14,15 epoxide derivatives

N-n-butyl, N-methyl-ll-O'^β-dibenzoyl-H'/lS'-epoxy-estra l'^ ^' dOtrien- 7α-yl) undecanamide ("EM 180") and ("EM 181")

BzCt, C ₅ H ₅ N r.t.

CPBA, CH2Cl2 r.t. 50

- 158 -

EXAMPLE 24 Scheme 24

N-n-butyl, N-methyl-ll-(3M7α-dihydroxy estraWdO ^' ) trien-7α-yl) undecanamide ("EM 187")

EXAMPLE 25 Scheme 25

N-n-butyl, N-methyl-ll-(6'-hydroxy-2'-(4 ^* '-hydroxyphenyl)-3 ^, -ethyl-indol-N,-yl) undecanamide (104)

The starting material 101 has been synthesized as described by Von Angered et al., J. Med. Chem. 27: 1439-1447, 1984.

103 104

EXAMPLE 26 (Scheme 26)

110

EXAMPLE 27 Scheme 27

N-n-butyl, N-methyl-ll-[4,4 ^, -(l,2-diethyl-l,2-ethanydyl) bis-phenol-3-yl) undecanamide (115)

hexestrol

OCO Etz. KzCOa CHaCOCH-.

Other sex steroid activity inhibitors in accordance with the invention may be synthesi2ed 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.

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 closed third ring which includes, in its sides the ethenyl double bond and one side of one of the phenyl groups. The closed third ring is exemplified by the "B" ring, for example, of formulas XX and XXI below:

Preferred side chains, which are believed to help provide antagonistic or inhibitory characteristics, include the R ^ϊ (B-R ² -] L-G and A ^* [Y-A"] _u XR ₂ i side

chains discussed and defined above. Preferred additional substituents to the nudeus 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 -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 C ₁ -C ₃ hydrocarbon such as methyl, ethyl or propyl. Generally, this substitution location (e.g. the atom receiving R ⁶ of formula XX or G ³ of formula XXI) is the atom which is both (A) one atom away from the A-ring and (B) one of the two atoms which receives the optional double bond when it is present. In contrast, the preferred side. chain (e.g. RKB- R ² )LG of formula XX and R ¹⁰⁰ LG ¹ G ² of formula XXI) is preferably substituted, as shown in formulas XX and XXI, at the atom which is both (A) one atom away from the optional pi bond and (B) one atom away from Z. It is believed that a 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 D substantially enhances the effectiveness of an inhibitor.

While both the A and D rings may be unsubstituted, certain embodiments indude hydroxy substitution on one or both of the A and D rings (espedally 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

would be converted to hydroxy in vivo after the pharmaceutical is administered to a patient. Such substituents indude, for example, methoxy, ethoxy or esters.

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

- 170

Non-limiting examples of synthesis of representative inhibitors are set forth below.

EXAMPLE 28 scheme 28

β-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 (20 ml). The mixture was heated at reflux during the night, cooled, 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 MgS0 ₄ 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 R^I (CH ₃ 1 ,85 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 MgS0 ₄ and the solvent was removed under reduced pressure and the compound 2 (Rc=CH ₃ ) was obtained in the yield of 80 %.The action of MeOCOCN (0.1 ml, 10 min at -78°C) on the compound 2 (R _C =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 mi ) and HMPA (0.15 ml) yield in 83% the compound 3 (R^CHa). To this compound (290 g) 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°C and then I(CH ₂ )eOTBDMS (657 mg, 1.9 mmol) was added and the mixture heated at reflux 6h.The extraction with ethyl acetate yield the compound 4 (n=8, R _C =CH ₃ ) (98%).The decarboalkoxylation with lithium bromide in pyridine yield the compound 5 (n=8, R _C =CH ₃ ) which was treated by

the Grignard's reagent of 3-bromo-anisole followed by acidic treatmen The resulting compound 6 (n=8, Rc=CH ₃ ) was converted into amide 7 (n=8, Ra=CH3, R _b = H _{9 r} R _c =CH ₃ ) Thus a cooled solution compound 6 (n=8, Rc=CH3> (700 mg) in acetone (17 ml) was added Jones' reagent (8N-chromic acid 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 acid 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 μl, 136 mmol) and isobutylchlorofoπnate (280 μl, 2.1 mmol). After 40 minutes, N-methylbutylamine (13 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 HQ, saturated sodium bicarbonate solution and water (3X), 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 g). Elution with a mixture of hexane-ethyl acetate gave the amide 7 (63%) The removal of the protection into compound 8 (EM 736, n=7, R _a =CH ₃ , R =C- ₄ H ₉ , R _c =CH ₃ ) is performed with pyridine-HCl.

TABLE1

EXAMPLE 29

Scheme 29

TBDMS: t-butyl dimethylsilyl MOM: Methyloxymethyl

The compound 6 (n=9, R _c =CH ₃ ) is deprotected with pyridine-HCl and the phenol function is selectively protected as MOM derivative by treatment with CH3 OCH ₂ CI in pyridine. The esterification of the alcohol with PTSC1 in pyridine gives the compound 9 (n=9, Rc =CH ₃ ) which is transformed in sulfide 10 (ns9,R _a =C5H ₆ F ₅ , Rc =CH ₃ ) with sodium hydride and Ra SH. The oxidation with mCPBA and the acidic hydrolysis gives the sulfoxide 11 (EM 732, n=9, R _a =CsH ₆ F ₅ , Rc=CH ₃ ) .

TABLE 2

EXAMPLE30 Scheme 30

lone 12

β-tetralone is alkylated with (MeO^CO and sodium hydride in THF at reflux followed the reaction of sodium hydride and RI (CH ₃ I) in THF and HMPA. The action of MeOCOCN in presence of LDA yield the compound 12 (R _C =CH ₃ ) which is alkylated into compound 13 (R _C =CH ₃ ) by I H4OTHP 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 (C ^ ₂ NC5H1 ₀ in presence of K2CO3) gives the ketone 14 (R _a R _b =CsHιo) which is transformed into compound 15 (R _a Rb=CsHιo) by reaction with the Grignard's reagent of Br C- ₆ H ₄ OTMS in ether. The acidic hydrolysis gives the compound 16 (EM 431, R _a Rb=CsHιo).

TABLE 3

EXAMPLE 31 Scheme 31

18

The compound 17 (n=6, R _{a b} =- ₅ Hιo-, Rc=CH ₃ ) is obtained by the reaction of HNR _a Rb and sodium hydride on the compound 9. The addic hydrolysis gives the compound 18 (EM 473, n=6, R _a Rb=-C5Hκ Rc=CH ₃ ).

EXAMPLE32 Scheme 32

The reaction of the o-tolualdehyde with the Grignard's reagent of BrCH ₂ C H ₄ θR _d (R _d = THP) followed by the oxidation of the resulting alcohol by PCC (pyridinum chlorochromate) gave the compound 19. The reaction with OHCC ₆ H ₄ θ(CH ₂ ) ₂ CsH o in presence of sodium hydride followed by bromination with N-bromosucdmide on presence of light in CCI4 gave the enone 20 (R _a R _b =-CsHιo-). The product 21 (R _a R _b =-CsHι ₀ -) is obtained by cydisation with (L-Bu) ₃ SnH and AIBN (2'-2'-azobisisobutyrordtrile) 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=-C5Hιo-, c=CH ₃ ) was obtained by cydisation with LDA followed by Grignard's reaction with alkyl bromide and addic removal of the protection.

TABLE 5

EXAMPLE 33 Scheme 33

EM349

The compound 26 ( R=H, Rc=H ) was reduced with sodium borohydride i ethanol and a mixture of resulting compound (300mg; 0.6mmol), 4-(2-chloro ethyDmorpholine hydrochloride (267mg; 1.4mmol) cesium carbonate (978mg; 3.0mmol) and potassium iodide (lOOmg; O.όmmol) in N,N-dimefhylformamide (10ml) was kept at 90°C and with stirring for lh. Water was added and the resulting mixture was extracted several times with a mixture of ether and ethyl acetate (1:1). Drying (MgSC> ₄ ) and removal of solvent gave a waxy material tha 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 (R=(CH ₂ ) ₂ C ₄ H ₈ θ, Rc=H ) (153mg; 41%).

A solution of above compound(153mg; 249μmol) in a mixture of acetic add (60ml) and water (6ml) was kept at 100°C for lOmin. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel (ethyl acetate: acetone; 3:1) to yield compound 28 (EM-349, R=(CH ₂ ) ₂ NCιH ₈ O, Rc=H XlOOmg 90%). (δ NMR; 300MHz; solvent CD ₃ OD; standard: TMS) 254 (4H; t J 4.5Hz cydo-N-CH ₂ -CH ₂ -0-CH ₂ -CH ₂ -) 2.73 (2H; t; 5.5Hz, 0-CH ₂ -CH ₂ -N) 3.66 (4H; t; J 4.5Hz; cydcv-N-CH2-CH ₂ -0-CH ₂ -CH ₂ -) 4.0 (2H; t J 5.5Hz; 0-CH ₂ -CH--N) 6.11 (IH; d; J 25Hz; CH Phenyl) 6.12 (IH; s; O-CH- Ph) 6.29 (IH; dd; J 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 (IH; d; J 8Hz; CH Phenyl) 6.95 (IH; s; HC=C) 7.25 (2H; d; J 85Hz; CH Phenyl) 731 (2H; d; J 8.5Hz; CH Phenyl)-

TABLE 6

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

TABLE 7

EXAMPLE 35 Same synthesis than example 33 in scheme 33, the compound 23 being replaced by the 2'-hydroxyphenylacetophenone

TABLE 8

EXAMPLE36

EM 350

Synthesis described in scheme 33

Thus the compound 27 (R=H, Rc=H) was reduced with sodium borohydride in ethanol and a mixture of resulting compound (300mg; 0.6mmol), chloroacetic add, piperidyl amide (242mg; 15mmol) and cesium carbonate (978mg;

3.0mmol) in N,N-dimethylformamide (10ml) was kept at 90°C and with stirring for Ih. Water was added and the resulting mixture was extracted several times with ether. Drying (MgSθ 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

A solution of above compound (127mg; 203μmol) in a mixture of acetic add (10ml) and water (1ml) was kept at 100°C for lOmin. 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 Hιo, Rc=H) (43mg; 46%). (δ NMR; 300MHz; solvent CD ₃ OD; standard: TMS) 1.4-1.7 (6H; m; cydo-N-CH ₂ -CH ₂ -CH ₂ -CH ₁ -CH ₂ -) 3.40 (2H; t; J 55Hz; cyclo-N-CH ₂ -CH ₂ -CH ₂ -CH ₂ -CH ₂ -) 3.49 (2H; t, J 5.5Hz; cyclo-N-CH ₂ -CH2- CH ₂ -CH ₂ -CH ₂ -) 4.67 (2H- s; 0-CH ₂ -CO-N) 6.11 (IH; d; J 2Hz; CH Phenyl) 6.13 (IH; s; O-CH-Ph) 6.29 (IH; dd; J 2Hz,8Hz; CH Phenyl) 6.69 (2H; d; J 8.5Hz; CH Phenyl)

6.79 (2H; d; J 85Hz; CH Phenyl) 6.94 (IH; d; J 8Hz; CH Phenyl) 6.95 (IH; s; HC=C) 7.25 (2H; d; J 8.5Hz; CH Phenyl) 7.32 (2H; d; J 85Hz; CH Phenyl). Mass Spectroscopy: M+ 457

28 (R= (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 (R=H, Rc=H) was reduced with sodium borohydridee in ethanol and a mixture of resulting compound (418mg; 0.84mmol), N- methyl,N-butyl chloroacetamide (342mg; 2.09mmol) and cesium carbonate (136g; 4.18mmol) in N,N-dimethylformamide (20ml) was kept at 90°C and with stirring for 12h. Water was added and the resulting mixture was extracted several times with ether. Drying (MgSCλi) 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 (138mg; 220μmol) in a mixture of acetic acid (10ml) and water (1ml) was kept at room temperature for lOmin. 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 357, R= CH ₂ CONMeBu, Rc=H) (38mg; 37%). (δ NMR; 300MHz; solvent: CD ₃ OD; standard: TMS) 0.85-1.0 (3H; m; N-CH ₂ -CH ₂ -CH ₂ - H3).1.2-1 5 (2H; m; N-CH ₂ -CH ₂ -CH ₂ -CH ₃ ) 1.4-1.65 (2H; m; N-CH ₂ -CH2-CH2-CH3) 2.87 and 2.96 (3H; 2s; N-CH ₃ ) 3.25-3.4 (2H; m; N-CH ₂ -CH2-CH2-CH3) 4.66 and 4.68 (2H; 2s; 0-CH ₂ - CO-N) 6.12 (IH; d; J 2.5Hz; CH Phenyl) 6.13 (IH; s; O-CH-Ph) 6.29 (IH; dd; J 25Hz,8Hz; CH Phenyl) 6.70 (2H d; J 85Hz; CH Phenyl) 6.78 and 6.79 (2H; 2d; J 85Hz; CH Phenyl) 6.94 (IH; d; J 8Hz; CH Phenyl) 6.95 (IH; s; HC=C) 7^5 (2H; d; J 85Hz; CH Phenyl) 732 (2H; d; J 85Hz; CH Phenyl). Mass Spectroscope M+ 459

-192-

EXAMPLE37

Scheme 34

NR _a Rb 34

33

EM 345

Synthesis described in scheme 34

A mixture of compound 24 (252g; 6.12mmol), the aldehyde 29 (n=7, R _a =Me,

R _b =Bu) (l.OOg; 4.08mmol) and piperidine (500μl) 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; 73) to yield the chalcone 30 (n=7, R _a =Me, Rb=Bu)

(620mg; 77% correded yield) and the unreacted starting material 24 (2.00g).

To a solution of the chalcone 30 (n=7, R _a =Me, R _b =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 (MgSθ 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 (n=7, R _a =Me, R _b =Bu, Rc=H)(300mg; 66%).

A solution of compound 31 (n=7, R _a =Me, R _b =Bu, R _c =H)(300mg; 482μmol) in a mixture of acetic acid (30ml) and water (3ml) was kept at 100°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 _a =Me, R =Bu, R _< ~H) (104mg; 48%). (δ NMR; 300MHz; solvent CD _Q3 ; standard: TMS) 0.85-1.0 (3H; m; N-CH ₂ -CH2-CH2-CH3).1.15-1.9 (16H; m; N-CH ₂ -CH ₂ -CH ₂ -CH ₃ and 0-CH-(CH ₂ ) ₆ -CH ₂ -CO-N) 2.3-2.4 (2H; m; CH ₂ -CO-N) 2.96 and 3.00 (3H; 2s; N-CH ₃ ) 3.28 and 3.40 (2H; 2m; N-CH2-CH2-CH2-CH ³ ) 5.15 (IH; dd; J 2Hz,10Hz; O-CH-CΗ ₂ ) 6.44 (IH; dd; J 2Hz,8Hz; CH Phenyl) 654 (IH; d; J 2Hz; CH Phenyl) 6.60 (IH; s; HC=C) 6.84 (2H; d; J 85Hz; CH Phenyl) 6.92 (IH; d; J 8Hz; CH Phenyl) 731 (2H; d; J 85Hz CHPhenyl). Mass Spectroscopy. M+ 451

TABLE 10

EXAMPLE38

Typical procedure for compounds 34 (EM 371, n=10, R _a =Me, R _b =Bu, Rc=H)

Synthesis described in scheme 34

To a solution of the amide 31 (n=10, R _a =Me, R _b =Bu, Rc=H) (lOOmg; 0.15mmol) in tetrahydrofuran (10ml) under reflux and with stirring was added a solution of lithium aluminium hydride (1M in tetrahydrofuran; 0.42ml; 0.42mmol) The resulting mixture was refluxed for a further 48h. 2N aqueous sodium hydroxide solution was added to the reaction mixture and the aqueous layer was extracted several times with ethyl acetate. The joined organic extracts were dried (MgSθ ₄ ) and solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (hexanes: acetone; 1:1) to yield the amine 33 (n=10, R _a =Me, R _b =Bu, R _< =H) (60mg; 62%).

A solution of above compound (60mg; 93μmol) and pyridinium p- toluenesulfonate (46mg; 185μmol) in methanol (10ml) was refluxed for 12h. 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, R _a =Me, R _b =Bu, R _< ~H) (30mg; 67%) (δ NMR; 300MHz; solvent: CD3OD; standard: TMS)0.93 (3H; t; J 7.5Hz; N-CH ₂ -CH ₂ -CH ₂ -C.H3).1.15-1.85 (24H; m; N-CH ₂ -CH ₂ -CH ₂ -CH ₃ and CJ-CH-(CH ₂ ιo-CH ₂ -N) 2.24 (3H; s; N-CH3) 235-2.45 (4H; m; CH ₂ -N-CH ₂ ) 5.19 (IH; dd; J 2.5Hz,10Hz; O-CH-CH ₂ ) 6.26 (IH; d; J 2.5Hz; CH Phenyl) 633 (IH; dd; J 2.5Hz,8.5Hz; CH Phenyl) 6.59 (IH; s; HC=C) 6.78 (2H;

d; J 8.5Hz; CH Phenyl) 6.88 (IH; d; J 8Hz; CH Phenyl) 7.30 (2H;.d; J 8.5Hz; CH Phenyl).

TABLE 11

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 eliminated-

TABLE 12

-200-

EXAMPLE40

35 36

4'-hydroxy-2-(4-benzoyloxyphenyl) acetophenone 35 prepared from the commerdal desoxyanisoin (250 g)(hydrolysis of the methoxyl group with pyridine-HCl at 220 °C followed by the treatment with 120 ml of benzoyl chloride and 20g of dimethylaminopyridine in 21 of methylene diloride 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 acid (10 g) in 21 of methylene chloride at 0 °C.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 commerdal thiosalicylic add 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 HCl and cesium carbonate in N,N-dimethylformamide (20 ml) is kept at 90°C and with stirring for 12h, extracted several times with ether. Drying (MgS0 ₄ ) and removal of solvent under reduced pressure gives a gummy residue that is purified by chomatography on silica gel (hexanes: ethyl acetate; l:l).The resulting compound in methanol (2 1) is heated at reflux with

p-toluenesulfonic add (5 g) for 3 h, Extraction with ethyl acetate and flash chromatography with silica gel (hexanes: ethyl acetate) gives the compound 39 (EM 547) (68 g) of wich structure was determined by spectroscopic means.

TABLE 13

EXAMPLE 41

Same synthesis than the synthesis described in sheme 35 except the starting coumpound 35 is HO H4CO(CH2)nCONRaRb

TABLE 14

EXAMPLE42 Scheme 36

46

The compound 40 obtained by acylation and methylation of the commercial m- anisidine is treated in a Friedel-Craft reaction with the 4-methoxyphenylacetoyl chloride 41 and AICI 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 by the method described in example 33. The resulting compound 45 is treated with methylmagnesium iodide in THF followed by pyridine-HCl treatment to give compound 46 (EM 821, R _a R = sHιo, Rc=CH ₃ ).

TABLE 15

EXAMPLE43

Scheme 37

52 53

The 4-methoxyphenylglycine 47 (prepared from commercial 4- hydroxyphenylglydne) is treated with (Cθα)2.The resulting add chloride 48 is used in a Friedel-Craft reaction on the 3-methoxybenzyl alcohol 49 in using aluminiumchloride as catalyst. The coupled compound 50 is treated with methyl magnesium iodide in THF followed by the treatment with p- toluenesulfonylchloride in pyridine and CH2 C I 2 and dimethylaminopyridine.The resulting compound 51 is heated and reduced with LiAlH4 into the compound 52. The alkylation with NaH and Br(CH2)ιoCONC4H8 in DMF gives the compound 53 (EM 877,

EXAMPLE 44

Same synthesis than the synthesis described in scheme 37. The compound 53 is reduced with LiAlH-4 in THF.

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 daims.