Title of Invention

2-ALKOXYESTRADIOL ANALOGS AND PHARMACEUTICAL PREPARATIONS

Cross-Reference to Related Applications [0001] N/A

Statement Regarding Federally Sponsored Research or Development [0002] N/A

Background of the Invention

[0003] 2-Methoxyestradiol is an endogenous mammalian metabolite formed by the sequential biochemical hydroxylation and methylation of the natural hormone Estradiol (Breuer, H. et al., Naturwissenschaften 12, pp. 280-281 (I960)). Certain 2- alkoxyestradiols have been discovered to have antitumor activity (U.S. Pat. No. 6,136,992; U.S. Pat. No. 6,054,598; U.S. Pat. No. 6,051,726; U.S. Pat. No. 5,892,069; U.S. Pat. No. 5,661,143; U.S. Pat. No. 5,504,074; WO 95/04535) (incorporated herein in their entirety by reference). 2-Methoxyestradiol (2-ME2) is one such 2-alkoxyestradiol exhibiting antitumor activity (U.S. Pat. No. 5,892,069; U.S. Pat. No. 5,661,143; U.S. Pat. No. 5,504,074; WO 95/04535).

[0004] Studies have demonstrated that the mechanism responsible for the antitumor activity exhibited by certain 2-alkoxyestradiols, including 2-ME2, includes interference with or prevention of cell mitosis, the multi-step process that precedes cell division and replication (Alberts, B. et al., The Cell, pp. 652-661 (1989); Stryer, L., Biochemistry (1988)). For example, some 2-alkoxyestradiols have been shown to inhibit the replication of certain cancer cells by interfering with microtubule formation and function (Seegers, J. C. et al., J. Steroid Biochem. 32, pp. 797-809 (1989); U.S. Pat. No. 6,136,992; U.S. Pat. No. 6,054,598; U.S. Pat. No. 6,051,726; U.S. Pat. No. 5,892,069; U.S. Pat. No. 5,661 ,143; U.S. Pat. No. 5,504,074; WO 95/04535). Microtubules facilitate and make possible, among other things, chromosome and organelle movement and segregation during cell mitosis (Stryer, L., Biochemistry (1988)). Preventing or interfering with microtubule formation and function leads to mitotic arrest and frequently to apoptosis. In addition to cancer, many diseases are characterized by undesirable cell proliferation, and the value of compounds and methods that prevent such undesirable cell proliferation is of great importance to the treatment of such diseases. Microtubule formation and function is also critical to cell maintenance, locomotion and the movement of specialized cell structures such as cilia and flagella (Stryer, L., Biochemistry (1988)).

[0005] To function properly, cilia and flagella require proper tubulin polymerization (U.S. Pat. No. 6,162,930). Certain 2-alkoxyestradiols are known to inhibit tubulin polymerization or to cause the formation of tubulin polymer with altered morphology and stability properties (U.S. Pat. No. 6,136,992). By interfering with normal microtubule dynamics, such compositions may be used to treat those diseases characterized by abnormal proliferation.

[0006] Certain 2-alkoxyestradiols, including 2-ME2, have also been demonstrated to act as antiangiogenic agents (Fotsis et al., Nature 368, pp. 237-239 (1994); U.S. Publication No. 20050014737, U.S. Pat. No. 6,136,992; U.S. Pat. No. 6,054,598; U.S. Pat. No. 6,051,726; U.S. Pat. No. 5,892,069; U.S. Pat. No. 5,661,143; U.S. Pat. No. 5,504,074; WO 95/04535). Such antiangiogenic activity is likely due to the arrest of endothelial cell mitosis and the consequent prevention of endothelial cell proliferation. 2- alkoxyestradiols exhibiting antiangiogenic activity may be used to treat diseases in which angiogenesis plays an important role. Inducing mitotic arrest and preventing angiogenesis will cause tumors to shrink, and the combination of these methods will provide significant advantages over current anticancer therapies. 2-alkoxyestradiols in murine models have been shown to be orally active, and to exhibit no appreciable toxicity at therapeutically effective doses (Fotsis et al., Nature 368, pp. 237-239 (1994)). [0007] Certain novel 2-alkoxyestradiols and derivatives of 2-alkoxyestradiols having antiproliferative and antiangiogenic activity were described in U.S. Patents 6,593,321 and 6,852,710 (incorporated herein in their entirety by reference). Compounds disclosed and tested include (i) 2-methoxyestra-l,3,5(10),15-tetraen-3,17β-diol, (ii) 2-methoxyestra- l,3,5(10),14-tetraen-3,17β-diol, (iii) 2-methoxyestra-l,3,5,(10),7tetraen-3,17β-diol, and (iv) 2-methoxy-3,15α,16α,17β-tetrahydroxyestra-l,3,5(10)-trien e 15,16-acetonide. The synthesis of several ring D modified 2ME2 analogs has been reported (Rao et al., Steroids 2002;67: 1079- 1089; Tinley et al., Cancer Res 2003;63:1538-1549) and the authors observed that by introducing an additional unsaturation in ring D such as a 14-dehydro or a 15-dehydro derivative, the in vitro antiproliferative activity could be considerably enhanced relative to the parent 2ME2 compound. Additionally, the 17-hydroxy analogs are more active in vitro than their 17-oxo counterparts. It is not known whether these structural modifications will enhance the binding affinity towards tubulin or decrease the rate of metabolic inactivation.

[0008] There is still a general need, however for additional antiproliferative and antimitotic chemical compounds that provide therapeutic advantages over those compounds currently available.

Brief Summary of Invention

[0009] This disclosure arises in part from the concept that the bioavailability of 2- methoxyestradiol is significantly decreased by metabolic oxidation of the 17-hydroxy group to give the inactive 2-methoxyestrone. Further metabolic inactivation of 2- methoxyestradiol may occur with conjugation of the 3- and/or 17-hydroxy groups with sulfate or glucuronic acid giving rise to derivatives that are more prone to excretion. The structural modifications disclosed herein are designed to prevent oxidation or conjugation at the 17-position and conjugation at the 3-position. It is contemplated that the specific modifications presented herein decrease metabolic inactivation while retaining the antiproliferative effects of the parent compound.

[00010] The syntheses and antimitotic activity of several novel 18a-homo-analogs of 2-methoxyestradiol are described. Structural modifications of the parent 2-methoxy- 18ahomoestradiol include introduction of unsaturation in the D-ring and methylation of the 17-OH. Additionally, the syntheses and antimitotic activity of several novel analogs of 2-methoxyestradiol are described with structural modifications that include ring-D homologation, aromatization of the six membered ring-D to a chrysine type molecule with alkyl substituents, and introduction of unsaturation in five-membered ring-D along with substitution of alkyl and ethynyl groups for the 17β-hydroxy function. Such analogs are contemplated to have superior antiproliferative activities compared to 2- methoxyestradiol. [00011] Certain embodiments of the disclosure are compounds of formula I :

[00012] or a pharmaceutically acceptable salt or ester thereof, wherein R is O-alkyl (C _{1- 6} ); R ² is OH, SO ₃ NHR ⁵ , with R ⁵ being hydrogen or alkyl (C _1-6 ), or CONHR ⁶ , with R ⁶ being hydrogen or alkyl (Ci _-6 ); R ³ is alkyl (C _1-6 ); and R ⁴ is H, OH, or OR ⁷ , with R ⁷ being alkyl (C _1-6 ). [00013] Certain embodiments are compounds of formula II:

[00014] or a pharmaceutically acceptable salt or ester thereof, wherein R is O-alkyl (Ci- ₆ ); R ² is OH, SO ₃ NHR ⁵ , with R ⁵ being hydrogen or alkyl (C _1-6 ), or CONHR ⁶ , with R ⁶ being hydrogen or alkyl (Ci _-6 ); R ³ is alkyl (Ci _-6 ); and R ⁴ is H, alkyl (Ci _-6 ), C≡R ⁷ with R ⁷ being H or alkyl (C _1-6 ), CN or COR ⁸ , with R ⁸ being H, alkyl (C _)-6 ), OH, O-alkyl (C _1-6 ), NH ₂ , or NH-alkyl (Ci _-6 ). [00015] Certain embodiments are compounds of formula III:

[00016] or a pharmaceutically acceptable salt or ester thereof, wherein R ¹ is O-alkyl (Ci- ₆ ); R ² is OH, SO ₃ NHR ⁶ , with R ⁶ being hydrogen or alkyl (C _{1 -6} ), or CONHR ⁷ , with R ⁷ being hydrogen or alkyl (Ci _-6 ); R ³ is alkyl (Ci _-6 ); R ⁴ (stereochemistry unspecified) is H, OH, O-alkyl (C _1-6 ), or SO ₃ NHR ⁸ with R ⁸ being hydrogen or alkyl (C _1-6 ), R ⁵ is H, OH, O- alkyl (Ci _-6 ), SO ₃ NHR ⁹ , with R ⁹ being hydrogen or alkyl (Ci _-6 ). [00017] Certain embodiments are compounds of formula IV:

[00018] or a pharmaceutically acceptable salt or ester thereof, wherein R ¹ is O-alkyl (C- ₆ ); R ² is OH, SO ₃ NHR ⁶ , with R ⁶ being hydrogen or alkyl (C _1-6 ) or CONHR ⁷ , with R ⁷ being hydrogen or alkyl (Ci _-6 ); R ³ is hydrogen, alkyl (Ci _-6 ), OH, O-alkyl (Ci _-6 ), SO ₃ NHR ⁸ with R ⁸ being hydrogen or alkyl (Ci _-6 ), or CONHR ⁹ , with R ⁹ being hydrogen or alkyl (C _-6 ); R4 is hydrogen, alkyl (Ci _-6 ), OH, O-alkyl (Ci _-6 ), SO ₃ NHR ¹⁰ with R ¹⁰ being hydrogen or alkyl (Ci _-6 ), or CONHR ¹¹ , with R ¹¹ being hydrogen or alkyl (Ci _-6 ); and R ⁵ is hydrogen, alkyl (C _{1 -6} ), OH, O-alkyl (C _1-6 ), SO ₃ NHR ¹² with R ¹² being hydrogen or alkyl (Ci _-6 ), or CONHR ¹³ , with R ¹³ being hydrogen or alkyl (C _-6 ).

[00019] Certain embodiments are compounds of formula V:

[00020] or a pharmaceutically acceptable salt or ester thereof, wherein R ¹ is O-alkyl (C- ₆ ); R ² is OH, SO ₃ NHR ⁴ , with R ⁴ being hydrogen or alkyl (C _-6 ) or CONHR ⁵ , with R ⁵ being hydrogen or alkyl (Ci _-6 ); and R ³ is alkyl (C _1-6 ). [00021] Certain embodiments are compounds of formula VI:

[00022] or a pharmaceutically acceptable salt or ester thereof, wherein R ¹ is O-alkyl (Ci- ₆ ); R ² is OH, SO ₃ NHR ⁵ , with R ⁵ being hydrogen or alkyl (C _1-6 ) or CONHR ⁶ , with R ⁶ being hydrogen or alkyl (Ci _-6 ); R ³ is alkyl (Ci _-6 ); and R ⁴ is OH, alkyl (Ci _-6 ), or SO ₂ NHR ⁷ , with R ⁷ being hydrogen or alkyl (Ci _-6 ). [00023] Certain embodiments are compounds of formula VII:

[00024] or a pharmaceutically acceptable salt or ester thereof, wherein R ¹ is O-alkyl (Ci- ₆ ); R ² is OH, SO ₃ NHR ⁵ , with R ⁵ being hydrogen or alkyl (Ci _-6 ) or CONHR ⁶ , with R ⁶ being hydrogen or alkyl (Ci _-6 ); R ³ is alkyl (Ci _-6 ); and R ⁴ is OH, O-alkyl (C _]-6 ), or SO ₃ NHR ⁷ , with R ⁷ being hydrogen or alkyl (Ci _-6 ). [00025] Certain embodiments are compounds of formula VIII:

[00026] or a pharmaceutically acceptable salt or ester thereof, wherein R ¹ is O-alkyl (Ci- ₆ ); R ² is OH, SO ₃ NHR ⁴ , with R ⁴ being hydrogen or alkyl (Ci _-6 ) or CONHR ⁵ , with R ⁵ being hydrogen or alkyl (C _1-6 ); and R ³ is alkyl (Ci _-6 ). Certain embodiments are compounds of formula IX:

[00027] or a pharmaceutically acceptable salt or ester thereof, wherein R ¹ is O-alkyl (Ci- ₆ ); R ² is OH, SO ₃ NHR ⁵ , with R ⁵ being hydrogen or alkyl (Ci _-6 ) or CONHR ⁶ , with R ⁶ being hydrogen or alkyl (Ci _-6 ); R ³ is alkyl (Ci _-6 ); and R ⁴ (stereochemistry unspecified) is OH, O-alkyl (Ci _-6 ), COR ⁷ with R ⁷ being hydrogen, alkyl (Ci _-6 ), OH, O-alkyl (Ci _-6 ), NH ₂ , or NH-alkyl (C, _-6 ), or SO ₃ NHR ⁸ , with R ⁸ being hydrogen or alkyl (Ci _-6 ). [00028] Any of the disclosed compounds as described above can be included in a pharmaceutical formulation or dosage form as described herein or as commonly known in the art. As such the compounds may be mixed with or dissolved in a pharmaceutically acceptable carrier in liquid or solid form as appropriate.

[00029] Throughout this disclosure, unless the context dictates otherwise, the word "comprise" or variations such as "comprises" or "comprising," is understood to mean "includes, but is not limited to" such that other elements that are not explicitly mentioned may also be included. Further, unless the context dictates otherwise, use of the term "a" may mean a singular object or element, or it may mean a plurality, or one or more of such objects or elements.

Brief Description of the Drawings

[00030] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be

better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[00031] FIG. 1 is a schematic of the syntheses of compounds 2-11.

[00032] FIG. 2 is an alternate synthesis of 2-Methoxy-18a-homoestra-l,3,5,(10)-trien-

17β-ol, compound 9.

[00033] FIG. 3 is a schematic of the syntheses of ring D-unsaturated 2-Methoxy-18a- homoestra-l,3,5,(10)-trien-3-ol derivatives.

[00034] FIG. 4 is a schematic of the synthesis of 2-Methoxy-3-hydroxy-18a-homoestra- l,3,5,(10),16-tetraene.

[00035] FIG. 5 is the molecular structures of compounds 36, 37, and 38, which have been observed with 13 -ethyl gauche g ⁺ conformation.

[00036] FIG. 6 shows the observed conformations of 18a homosteroids including the trans (left) and gauche (right) conformations.

[00037] FIG. 7 is the MM3 energy profile for the rotation of the 13 -ethyl group of compounds 9 and 24.

[00038] FIG. 8 is the MM3 energy profile for the rotation of the 13 -ethyl group of compounds 9 and 28.

[00039] FIG. 9 is the molecular structures of the compounds compared in Table 3.

[00040] FIG. 10 is a schematic of the syntheses of compounds 2a - 12a.

[00041] FIG. 11 is a schematic of the syntheses of 2-methoxy-17-methyl and 2- methoxy-17-ethynyl steroids with unsaturated ring D.

[00042] FIG. 12 is a schematic of the synthesis of a Dimethylhexahydrochrysine analog.

[00043] FIG. 13 is a schematic of the synthesis of 2-methoxyestra-l,3,5(10),14,15- pentaen-3-ol.

[00044] FIG. 14 is a schematic of the synthesis of Dihydroxyhexahydrochrysine analog.

Detailed description

[00045] In steroid metabolism, 17β-hydroxysteroid dehydrogenases (17β-HSD) play a major role in oxidization of a 17β-hydroxy function to an oxo group or the reduction of a 17-oxo group to a 17β-hydroxy derivative. Since it has been observed that the 17-hydroxy derivatives of 2ME2 are more active than their corresponding 17-oxo derivatives, it is desirable to minimize the activity of 17β-HSD on the oxidation of a 17β-hydroxy steroid to a 17-oxo compound. The present disclosure addresses this by incorporation of a sterically bulky substituent next to the 17β-hydroxy function and also protecting the 17β-hydroxy

group as a methyl ether which cannot be readily hydrolyzed by the enzyme hydrolase. 2ME2 has a methyl group at C- 13 position adjacent to the 17β-hydroxyl group. Replacing the C- 13 methyl group with a sterically bulkier ethyl group, it is contemplated that the 17β- hydroxyl function is protected from metabolic deactivation. Examples of 2ME2 derivatives with C- 13 ethyl substituents have been synthesized for evaluation of cytotoxic activity in multiple tumor cell lines.

[00046] One possible explanation for the effects of the changes in the 2ME2 structure on biological activity could lie in the observation of crystal structural data reported for certain 18a-homo steroids containing unsaturation in ring D. van Geerestein el α/(Acta Cryst C 1987;43:2402 - 2405; Acta Cryst C 1988;44:329 - 332) and Eckle et α/,(Liebigs Ann Chem 1988:199 - 202) have observed an unusual 18a-methyl orientation in the crystal structure of Gestodene (36) and 11 -methyl enegestodene (37). Normally, the observed orientation of the ethyl group in 18a-homo steroids with saturated D-rings is trans with respect to the C/D ring junction. However, crystal structures of compounds (36) and (37) were observed with the C 14-Cl 3 -C 18-Cl 8a bond in the gauche g ⁺ configuration with the 18a-methyl group directly over the D-ring. A similar observation was made by Chekhlov et al (Bioorg Khim 1983;9:978-985) for rac-3-methoxy-18a-homoestral-l,3,5(10),8,14-pentaen-17-one (38). These compounds along with an illustration of the conformational difference are shown in Figs. 5 and 6.

[00047] In order to investigate this possibility for compound (24) and (28), conformational energy profiles were calculated for these compounds along with the parental compound (9) using the MM3 forcefield (Lii and Allinger, J Comput Chem 1998;19:1001-1016 and references therein) implemented in PC-Model. The results are shown in Figs. 7 and 8. [00048] While the expected trans conformation for compound (9) is favored by ~ 2.5 Kcal/mol, these calculations indicate the gauche g ⁺ conformation is favored for the δ compound (24) by about 1.5 Kcal/mol and both conformations are about equally populated for the δ ¹⁵ -compound (28). Table 1 summarizes the results of similar calculations carried out for all the 18a-homo derivatives tested in this study.

[00049] Experimental evidence in support of these calculations can be seen in the chemical shift data for the Cl 8a methyl group for the 18a-homosteroids in this investigation. Table 3 compares these values along with the data for the corresponding 13 -methyl compounds (34), (39), and (35). It can be seen from these data that there is an increase in upfield shifts in the 18a-methyl for the sequences (9)-(28)-(24) and (ll)-(31)-(27) which is opposite of the

trend observed for the 13-methyl compounds (34)-(39)-(35). This could be explained by increased shielding of the methyl group by the double bond caused by an increased population of the gauche g ⁺ conformation as predicted by the calculations in Figs. 6 and 7 and Table 1.

[00050] Without limiting the present disclosure to a particular theory, the net effect of an increased population of the gauche g ⁺ conformations for compounds (24), (27), (28) and (31) would be a decrease in accessibility of the beta-face of the D-ring for interaction with tubulin. This could explain the decrease in biological activities observed for these compounds compared to (9) and (11). This is in agreement with the observation that for 16- substituted or 15,16-disubstituted 2ME2 analogs, the β-isomer is usually less active than the α-isomer.

[00051] While the Ring-D modifications carried out here with the introduction of unsaturation and absence of a hydroxyl function can increase activity compared to 2ME2, clearly other factors play a role. In a comparison of the MM3 minimized molecular structures of compounds 2ME2, 15a, 17a, 24a, and 30a whereby the A-ring of compounds 15a, 17a, 24a, and 30a are superimposed on the A-ring of 2ME2 via the best root mean square fit, the active compounds (17a, 24a, 30a) have a D-ring that is either in line with that of 2ME2 or projects towards the α-side of the molecule. In contrast, the least active compound (15a) has a D-ring that projects above that of 2ME2, towards the β-side of the molecule. Other investigation disclosed herein has suggested that substituents on the β-side of the D-ring decrease antiproliferative activity. In the case of compound (15a) the δ ¹³ ' ¹⁷ bond projects the entire D-ring towards the βface of the molecule which could explain its decreased antiproliferative activity. Indications

[00052] The disclosed compounds can be used to treat diseases characterized by abnormal or undesired cell proliferation. Such diseases include for example, but are not limited to: abnormal stimulation of endothelial cells (e.g., atherosclerosis), solid tumors and tumor metastasese, benign tumors (e.g., hemangiomas, acoustic neuromas, neurofibromas, trachomas and pyogenic granulomas), vascular malfunctions, abnormal wound healing, inflammatory and immune disorders, Bechet's disease, gout or gouty arthritis, abnormal angiogenesis accompanying for example, rheumatoid arthritis, psoriasis, diabetic retinopathy, and other ocular angiogenic diseases such as retinopathy

of prematurity (retrolental fibroplasia), macular degeneration, corneal graft rejection, neuroscular glaucoma and Oster Webber syndrome.

[00053] Neoplasms which the disclosed compounds can be used to treat include, but are not limited to: mammary, small-cell lung, non-small-cell lung, colorectal, leukemia, lymphoma, melanoma, pancreatic, renal, liver, myeloma, multiple myeloma, mesothelioma, central nervous system including neuroblastoma, ovarian, prostate, sarcoma of soft tissue or bone, head and neck, esophageal, stomach, bladder, retinoblastoma, squamous cell, testicular, vaginal, and neuroendocrine-related, which includes thyroid, Hodgkin's disease and non-Hodgkin's disease neoplasms. [00054] In certain embodiments, the disclosed compounds can be used in combination with other treatment modalities, e.g. surgery or radiation therapy, and in combination therapy with other known chemotherapeutic or antineoplastic agents (e.g., alkylating agents, antimetabolites, antitumor antibiotics, antimitotics (e.g., vinca alkaloids and taxanes), hormones (e.g., tamoxifen), Selective Estrogen Receptor Modulators (SERMs), antibodies (e.g., Herceptin), and platinum coordination complexes, etc.). For example, the compounds of the present invention can be used in combination therapy with a vinca alkaloid compound, such as vinblastine, vincristine, Taxol™, etc.; an antibiotic, such as adriamycin (doxorubicin), dactinomycin (actinomycin D), daunorubicin (daunomycin, rubidomycin), bleomycin, plicamycin (mithramycin) and mitomycin (mitomycin C), etc.; an antimetabolite, such as methotrexate, cytarabine, azauridine, azaribine, fluorodeoxyuridine, deoxycoformycin, mercaptopurine, etc.; or a platinum coordination complex, such as cisplatin, carboplatin, etc. In addition, those of skill in the art will appreciate that the disclosed compounds can be used in combination therapy with other known chemotherapeutic or antineoplastic compounds.

[00055] Certain embodiments relate to methods of treating diseases associated with undesired angiogenesis, the methods including administering to a subject an anti- angiogenic compound of any of Formulas I- VIII, or a pharmaceutically acceptable salt or ester thereof, prodrug or precursor thereof, in admixture with one or more pharmaceutically acceptable carriers, diluents, or excipients, in a therapeutically effective amount.

[00056] The methods can be used to treat a variety of diseases, including diseases associated with undesired angiogenesis. Such diseases include those associated with corneal neovascularization including, but are not limited to, diabetic retinopathy,

retinopathy of prematurity, corneal graft rejection, neovascular glaucoma and retrolental fibroplasias, epidemic keratoconjunctivitis, Vitamin A deficiency, contact lens overwear, atopic keratitis, superior limbic keratitis, pterygium keratitis sicca, sjogrens, acne rosacea, phylectenulosis, syphilis, mycobacterial infections, lipid degeneration, chemical bums, bacterial ulcers, fungal ulcers, Herpes simplex infections, Herpes zoster infections, protozoan infections, Kaposi's sarcoma, Mooren ulcer, Teπϊen's marginal degeneration, marginal keratolysis, trauma, rheumatoid arthritis, systemic lupus, polyarteritis, Wegener's sarcoidosis, scleritis, Steven Johnson's disease and periphigoid radial keratotomy.

[00057] Diseases associated with retinal/choroidal neovascularization that can be treated with the disclosed methods include, but are not limited to, diabetic retinopathy, macular degeneration, sickle cell anemia, sarcoid, syphilis, pseudoxanthoma elasticum, Pagets disease, vein occlusion, artery occlusion, carotid obstructive disease, chronic uveitis/vitritis, mycobacterial infections, Lyme's disease, systemic lupus erythematosus, retinopathy of prematurity, Eales disease, Bechets disease, infections causing a retinitis or choroiditis, presumed ocular histoplasmosis, Bests disease, myopia, optic pits, Stargarts disease, pars planitis, chronic retinal detachment, hyperviscosity syndromes, toxoplasmosis, trauma and post-laser complications. Other diseases include, but are not limited to, diseases associated with rubeosis (neovascularization of the angle) and diseases caused by the abnormal proliferation of fibrovascular or fibrous tissue, including all forms of proliferative vitreoretinopathy, whether or not associated with diabetes. [00058] Diseases associated with chronic inflammation can also be treated. Diseases that can be treated with the disclosed methods include diseases with symptoms of chronic inflammation that include, but are not limited to, inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis, psoriasis, sarcoidosis and rheumatoid arthritis. Unwanted angiogenesis is a key element that these chronic inflammatory diseases have in common. The chronic inflammation depends on continuous formation of capillary sprouts to maintain an influx of inflammatory cells.

[00059] Other diseases that can be treated include endometriosis, hemangiomas, Osler- Weber-Rendu disease, or hereditary hemorrhagic telagiectasia, solid or blood borne tumors and acquired immune deficiency syndrome (AIDS).

[00060] The disclosed compositions and methods can also be used to treat diseases, other than cancer for example, in which normal tubulin polymerization and function

plays a role. Chagas' disease, for example, is caused by Trypanosoma cruzi, a flagellate protozoa which has a substantial protein composition containing tubulin both as a component of the subpellicular microtubule system and the flagellum. Chagas ¹ disease is characterized by lesions in the heart, alimentary tract and nervous system. The disease is the leading cause of myocarditis in the Americas. Inhibition of tubulin polymerization, crucial to the parasite's mobility, would provide an effective treatment. Indeed, the use of agents that selectively affect tubulin polymerization has precedence in the therapy of other parasitic diseases. The benzimidazoles are very effective antihelmenthic drugs, and the dinitroanilines have shown promise against Leishmania, a parasite closely related to Trypanosoma (U.S. Pat. No. 6,162,930). The disclosed compositions are contemplated to be used to contact such parasites or sites of parasitic infection and thereby treat the associated disease.

[00061] Certain embodiments relate to a method of treating fungal diseases, the method comprising administering a therapeutically effective amount of a compound of any of Formulas I- VIII, or a pharmaceutically acceptable salt or ester, prodrug or precursor thereof, in admixture with one or more pharmaceutically acceptable carriers, diluents, or excipients. The progression of fungal diseases has proven particularly susceptible to treatment by drugs that act by disruption of microtubule organization, as for example by cryptophycin (U.S. Pat. No. 6,180,679). The methods of the present invention may be used in controlling mycotic infections or in controlling a yeast infection, where controlling refers to slowing, stopping or interrupting the spread of the given infection and not necessarily to a complete and total elimination of the mycotic infection or yeast infection. Administration

[00062] The disclosed compositions can be provided in therapeutically effective amounts as physiologically acceptable formulations using known techniques, and these formulations can be administered by standard routes. In general, the compositions can be administered alone or in combination, and by topical, oral, rectal, intravenous, subcutaneous or intramuscular route. In addition, the compounds can be incorporated into biodegradable polymers allowing for sustained release, the polymers being implanted in the vicinity of where delivery is desired, for example, at the site of a tumor. Biodegradable polymers and their use are described in detail in Brem et al., J. Neurosurg. 74, 441-446 (1991), and are familiar to those skilled in the art.

[00063] As will be appreciated by one skilled in the art, the dosage of the compositions will depend on the condition being treated, the particular compound used, the type and severity of the disease or malady, and other clinical factors such as weight, sex, age and condition of the patient, the patient's tolerance to drugs and/or treatment, and the route of administration. Those skilled in the art will be able to determine the appropriate dosages depending on these and other factors. Typically, a therapeutically effective amount of a compound can range from about 1 mg per day to about 1000 mg per day for an adult human individual. For oral administration to human adults, a dosage of 0.01 to 100 mg/kg/day, preferably 0.01-1 mg/kg/day, is generally a therapeutically effective amount. [00064] The formulations include those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intraocular, intratracheal and epidural) administration. The formulations can conveniently be presented in unit dosage form and may be prepared by conventional pharmaceutical techniques. Such techniques may include the step of bringing into association the active ingredient and the pharmaceutical carrier(s) or excipient(s). Pharmaceutical carriers or excipients can contain inert ingredients which do not interact with the compound, or ingredients that do interact with the compound but not in a fashion so as to interfere with the desired effect. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

[00065] Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil emulsion and as a bolus, et cetera. The oral formulations can be designed for immediate release in a gastric fluid environment or they can be designed for delayed or sustained release. The use of hydrophobic and hydrophilic polymers for coatings or drug matrices is known in the art and is contemplated for use in the described pharmaceutical formulations. Oral dosage forms can be prepared in which release is pH sensitive, to be released only at low or high pH, pH insensitive, released over a period of time of from 30 minutes to 1 to 4 hours, or even 10, 20, 30 or more hours, for example. In this way, release of the compositions can be targeted to a particular portion of the

digestive tract such as the gastric, ileal or colonic areas to provide for optimal absorption or stability of the active ingredient. The use of sustained release also allows the use of less frequent administrations, resulting in a more convenient protocol.

[00066] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface-active or dispersing agent. Molded tablets may optionally be coated or scored and may be formulated so as to provide a slow or controlled release of the active ingredient therein as described above. Oral dosage forms can also include capsules such as gel capsules that contain particles of therapeutic compositions in which either the particles or capsule can include coating layers.

[00067] Formulations suitable for topical administration in the mouth include lozenges comprising the ingredients in a flavored basis, usually sucrose and gum acacia or gum tragacanth; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and gum acacia; and mouthwashes comprising the ingredient to be administered in a suitable liquid carrier.

[00068] Formulations suitable for topical administration to the skin may be presented as ointments, creams, gels and pastes comprising the ingredient to be administered in a pharmaceutically acceptable carrier. A preferred topical delivery system is a transdermal patch containing the ingredient to be administered.

[00069] Formulations for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate.

[00070] Formulations suitable for nasal administration, wherein the carrier is a solid, include a coarse powder having a particle size, for example, in the range of 20 to 500 microns which is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nose. Suitable formulations for administration wherein the carrier is a liquid, for example, are nasal sprays or nasal drops including aqueous or oily solutions of the active ingredient.

[00071] Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known by those skilled in the art to be appropriate.

[00072] Formulations suitable for parenteral administration include aqueous and nonaqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described herein.

[00073] In addition, the compositions of the present disclosure may be administered in a formulation including liposomes in order to improve availability and to regulate dosage. The liposome may or may not form part of a targeted drug delivery system, for example in a liposome coated with a tumor-specific antibody. Such liposomes will be targeted to and taken up selectively by the site of interest (e.g., a tumor cell) Liposomes and emulsions are well known examples of delivery vehicles or carriers. Further, long- circulating, or stealth, liposomes may be employed (U.S. Pat. No. 5,013,556) (incorporated herein by reference).

[00074] Generally, such liposomes or other drug delivery systems typically have a targeting moiety, i.e., ligand, conjugated thereto that is specific for the target site of interest (e.g., tumor cell). For instance, some property (biochemical, architectural or genetic) of the tumor that is different from normal tissue can be exploited to concentrate the compounds of the present invention in, or at least near, the target tumor. Tumor vasculature, which is composed primarily of endothelial cells, is inherently different than normal differentiated vasculature. For example, the architecture of tumor vasculature is known to be leaky, and blood flow through them is mostly intermittent, with periods of perfusion and periods of occlusion and subsequent hypoxia. This aberrant microenvironment may be caused by and, in turn, leads to, additional differential gene expression in tumor vasculature relative to normal vasculature. This abnormal architecture and function, at the molecular level, is characterized by differences in surface markers in tumor microvessels relative to normal vessels and such differences can be exploited to target the liposome or other drug delivery system to the site of interest. Liposomes offer the added advantage of shielding the drug from most normal tissues.

When coated with polyethylene glycol (PEG) (i.e., stealth liposomes) to minimize uptake by phagocytes and with a tumor vasculature-specific targeting moiety, liposomes offer longer plasma half-lives, lower non-target tissue toxicity and delivery, and increased efficacy over non-targeted drug.

[00075] Other targeting strategies include, but are not limited to, ADEPT (antibody- directed enzyme prodrug therapy), GDEPT (gene-directed EPT) and VDEPT (virus- directed EPT). In ADEPT, the targeting of an inactive prodrug to a tumor mass is effected by an antibody against a tumor-associated marker. The enzyme milieu in or about the tumor transforms the prodrug into an active toxic agent that then acts on the tumor tissue. Similarly, differential gene expression or viral targeting at the tumor site is used to activate a prodrug into its active, toxic form in GDEPT and VDEPT, respectively. Other strategies include targeting differentially expressed genes, enzymes or surface markers that appear on, for example, tumor-associated vasculature to effect control of tumor progression or to other sites of interest (e.g., endothelial cells, TNF-. alpha., TNF- . alpha, receptor, etc.).

[00076] Additionally, standard pharmaceutical formulation techniques may be employed such as those described in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa (incorporated herein by reference). Additional methods of encapsulating compounds or compositions comprising the compound are known to those skilled in the art (Baker et al., "Controlled Release of Biological Active Agents", John Wiley and Sons, 1986).

[00077] Preferred unit dosage formulations are those containing a daily dose or unit, daily sub-dose, as herein above recited, or an appropriate fraction thereof, of the administered ingredient.

[00078] It should be understood that in addition to the ingredients specifically set forth above, the formulations can include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include flavoring and other agents. Chemistry

[00079] Melting points were determined on a Thomas-Hoover apparatus and are uncorrected. Nuclear magnetic resonance spectra were recorded on a General Electric GE-300 (300 MHz) spectrometer as deuterochloroform (CDCL ₃ ) solutions using tetramethylsilane (TMS) as an internal standard (δ = 0) unless noted otherwise. Infrared spectra were

recorded on Thermo-Nicolet model 370 FT-IR instrument equipped with an attenuated reflectance (ATR) accessory. Combustion analyses were performed by Midwest Microlabs Ltd. (Indianapolis, IN). 'Flash column' chromatography was performed on 32-64 uM silica gel obtained from EM Science, Gibbstown, New Jersey. 'Dry column' chromatography was performed on 70-230 mesh silica gel, also obtained from EM Science. Thin-layer chromatography (TLC) analyses were carried out on silica gel GF (Analtech) glass plates (2.5 cm x 10 cm with 250 uM layer and prescored). Most chemicals and solvents were analytical grade and used without further purification. Commercial reagents were purchased from Aldrich Chemical Company (Milwaukee, WI). Levonorgestrel was provided as a gift by Wyeth-Ayerst Research, CN 8000, Princeton, NJ 08534-8000.

[00080] The 3 -hydroxy- 17-ketone derivative (1) was prepared from Norgestrel according to Rao et al., Steroids 1994;59:621-627. The syntheses of compounds (2-11) are outlined in Fig. 1 and were based on the procedures previously developed for the 13 -methyl analogs (Rao and Cessac, Steroids 2002;67: 1065- 1070). Reduction of the ketone via sodium borohydride in ethanol/THF at room temperature overnight gave the diol compound (2) in 98% yield. Simple acetylation of this material with acetic anhydride in pyridine at room temperature for 48 hours gave the 3,17β-diacetate compound (3) in 80% yield. Treatment of the diacetate derivative (3) with zirconium (FV) chloride in dichloromethane at room temperature for 48 hours gave the 2-acetyl-3-hydroxy-17β-acetate compound (4) in 63% yield. This compound was then converted to the 3-benzyl ether derivative (5) in 87% yield by reaction with benzyl chloride in the presence of potassium carbonate in DMF at 65°C overnight. Conversion of this material to the diacetate derivative (6) was accomplished by reaction with 3-chloroperoxybenzoic acid and sodium phosphate in dichloromethane at room temperature for 48 hours in 79% yield. Simple base hydrolysis of the diacetate material (6) with sodium hydroxide in methanol/water gave the 2,17β-diol-3- benzyl ether compound (7) in 71% yield. Methylation of this material in THF with dimethyl sulfate in the presence of lithium hydroxide monohydrate at 60 ⁰ C overnight gave the 2- methoxy derivative (8) in 92% yield. Removal of the benzyl ether from compound (8) via catalytic hydrogenation with 5% palladium on carbon in ethanol at 40 psi gave compound (9) in 81% yield. Compound (8) was also converted to the 2,17β-dimethoxy derivative (10) in 64% yield by refluxing with solid sodium hydride in THF, followed by the addition of iodomethane. Finally, catalytic hydrogenation of compound (10) with 10% palladium on carbon in ethanol at 40 psi gave compound (11) in 94% yield.

[00081] An alternate synthesis of compound (9) is illustrated in Fig. 2 and is based on the procedure described in US Patent No. 6,488,419. Treatment of 2,17β-diol derivative (2) with chloromethyl methyl ether and N,N-diisopropylethylamine at 65 ⁰ C in THF overnight afforded the dimethoxymethyl ether (12) in 78% yield. This material was then reacted with n-BuLi in THF at -65 ⁰ C followed by the addition of trimethyl borate and sodium perborate to give the 2-hydroxy compound (13) in 84% yield. Methylation of this material in THF with dimethyl sulfate in the presence of lithium hydroxide monohydrate at 60 ⁰ C overnight gave the 2-methoxy derivative (13) in quantitative yield. This material was then hydrolyzed with 6M HCl in THF to give the 3,17β-diol derivative (9) in 72% yield. [00082] For the synthesis of ring D-unsaturated compounds (15-31), the general procedure reported in Rao et al, Steroids 2002;67:1079-1089, was followed and the synthetic scheme is presented in Fig. 3. The 3,17β-diol derivative (9) was then selectively acetylated in isopropanol with acetic anhydride in the presence of 2M KOH to give the 3-acetate derivative (15) in 91% yield. Oxidation of this material with Jones's reagent in acetone at 0 °C gave the 17-ketone compound (16) in 95% yield. Treating this material with triethylorthoformate, ethylene glycol and tosic acid in methylene chloride at room temperature overnight gave the 17-ketal (17) in 94% yield. Reaction of the 17-ketal (17) with phenyltrimethylammonium tribromide in THF at -5 °C afforded the 16α -bromo-17,17- ethylenedioxy compound (18) in 95% yield. Dehydrobromination by refluxing with potassium t~butoxide in xylenes gave a mixture of the δ ¹⁴ and δ ¹⁵ compounds (19 and 20) in 49% total yield. The mixture of unsaturated compounds (19 and 20) was reacted with tosic acid in 6:1 acetone/water at room temperature to give a mixture of δ ¹⁴ and δ ¹⁵ ketone derivatives (21 and 22) as well as the pure δ ¹⁵ compound (22), with a total yield of 49% . The mixture of isomers (21 and 22) was refluxed with isopropenyl acetate and tosic acid in acetic anhydride to give the pure δ ¹⁴ ' ¹⁶ -3,17-diacetate compound (23) in 46% yield. This material was then reduced with sodium borohydride in 1 :1 methanol/water at 0 ⁰ C to give the δ ¹⁴ -3,17B-diol (24) in 69% yield.

[00083] Selective acetylation of compound (24) in isopropanol with acetic anhydride in the presence of 2M NaOH gave the 3-acetate material (25) in 77% yield. The 3-acetate material (25) was then O-methylated with trimethylsilyl diazomethane in the presence of aqueous fluoroboric acid according to the procedure of Aoyama et al (Tetrahedron Letters 1990;38:5507-8) to give the 17β-methoxy compound (26) in 71% yield. Hydrolysis

of this material with potassium carbonate in 3:1 methanol/water at room temperature gave the 3-hydroxy compound (27) in 75% yield.

[00084] Reduction of the δ ¹⁵ compound (22) with lithium aluminum hydride in ether at 0 ⁰ C gave the δ ¹⁵ -3,17β-diol (28) in 37% yield. This material was then selectively acetylated in isopropanol with acetic anhydride in the presence of 2M NaOH to give the 3 -acetate derivative (29) in 61% yield. Methylation of this material with trimethylsilyl diazomethane in methylene chloride in the presence of aqueous fluoroboric acid (48%) at 0 ⁰ C gave the 17 ^β -methoxy compound (30) in 71% yield. Hydrolysis of this material with potassium carbonate in 3:1 methanol/water at room temperature gave the 3-hydroxy compound (31) in 76% yield.

[00085] The synthesis of the tetraene derivative (33) is presented in Fig. 4 and is based on the procedure of US Patent No. 5,793,571. Compound (16) was refluxed with p- toluenesulfonhydrazide in methanol in the presence of HCl for 16 hours to give the 3-hydroxy- 17-p-toluenesulfonylhydrazone derivative (32). This material was then treated with n-BuLi in THF at 0°C for 72 hours to give the δ ¹⁶ material (33) in 19% yield. [00086] The syntheses of the D-homo analogs of 2ME2 are outlined in Fig. 10 and are based on the Tiffeneau-Demjanov rearrangement as carried out by Avery et al (Steroids 1 990;55 :5964). Reaction of 2-methoxyestradiol (Ia) with sodium hydroxide and acetic anhydride in isopropanol selectively gave the 3 -acetate compound (2a) in quantitative yield. Oxidation of (2a) using Jones reagent in acetone at 0 ⁰ C afforded the 17-ketone derivative (3a) in 91 % yield. Hydrolysis of this material with sodium hydroxide in THF/methanol gave 2-methoxyestrone (4a) in quantitative yield. Treatment of compound (4a) with chloromethyl methyl ether and diisopropylethylamine in THF at 65 ⁰ C overnight afforded the 3-methoxymethoxy ether (5a) in 89 % yield. This material was then reacted with zinc iodide and trimethylsilylcyanide in chloroform at room temperature overnight. Analysis by ¹ H NMR indicated an incomplete reaction. The residue was re-reacted for four hours and purified by flash column. Analysis of the recovered material by ¹ H NMR indicated the loss of the 3-methoxymefhyl ether. The residue was re-protected by reaction with chloromethyl methyl ether and diisopropylethylamine at 65 ⁰ C overnight to give (6a) in 40 % overall yield. Reduction of the 17α-cyano group of (6a) was carried out with LiAlH ₄ in THF to give the 17a- aminomethyl-derivative (7a) in 75 % yield. Reaction of compound (7a) with sodium nitrite in aqueous acetic acid gave compounds (8a and 9a) in 61 % and 5.5 %

respectively. Treatment of compound (8a) with lithium tri-tørtbutoxyaluminum hydride solution in THF at 0 °C followed by hydrolysis with HCl gave products (10a and Ha) in 32 % and 10 % respectively. Finally compound (12a) was produced from (9a) in 43 % yield using the same conditions.

[00087] Stereochemistry for compounds (10a), (Ha) and (12a) were assigned based on the proton NMR chemical shifts for the 18-methyls and 17a-protons or 17-protons. This data is summarized in Table 2. According to Bhacca and Williams (Applications of NMR spectroscopy in organic chemistry. Illustrations from the steroid field. San Francisco. Holden-Day Inc., 1964:78), axial protons attached to hydroxyl-substituted carbons resonate at higher field than the corresponding equatorial protons in the epimeric alcohol. As can be seen from Table 2, this observation is consistent with the assignment of 17aβ- OH for compound (10a) and 17aα-OH for compound (Ha). These assignments agree with those made by Hillisch et al (US Application Publication No. US2006/0154985) for the corresponding compounds as the 3-sulfamate derivatives. The assignment of 17β-OH for compound (12a) is based on the strong downfield shift of the 18-methyl group due to 1,3-diaxial interaction with the 17β-axial OH.

[00088] The syntheses of D-ring unsaturated 2ME2 analogs (13a-17a) are outlined in Fig. 11. The 17-ketone (5a) was converted to the 17β-hydroxy derivative (13a) by reduction with sodium borohydride in 99 % yield. Compound (13a) was then treated with />-toluenesulfonic anhydride in pyridine at 0° C to give the 17β-tosylate product (14a) in 95 % yield. This material was then subjected to a Miescher-Kagi rearrangement following the procedure of Engel et al (J. Org. Chem. 1983;48: 1954-1966) by refluxing with ethyl magnesium bromide in benzene to give the 18-nor-13(17)-ene derivative (15a) in 79 % yield.

[00089] Compound (5a) was treated with trimethylsilylacetylene and butyl lithium at - 78 ⁰ C in THF/benzene then stirred at room temperature for 2 hours. Once TLC confirmed the formation of the lithium intermediate, methanesulfonyl chloride was added followed V2 hour later with pyridine and the mixture stirred at room temperature overnight. Subsequent extraction and purification gave a mixture of the 3 -hydroxy and 3- methoxymethoxy compounds (16a) which were used directly for the next reaction. This material was then treated with tetrabutylammonium fluoride at room temperature for 2 hours. Multiple purifications by flash column subsequent crystallization from acetone/hexanes gave the product (17a) in 6.4 % yield.

[00090] The syntheses of compounds (19a-24a) are outlined in Fig. 12. Compound (19a) was prepared as described by Vincze et al. (Steroids 1993;58: 220-224) by refluxing mestranol (18a) in formic acid. This material was converted to the free hydroxy derivative (20a) in 76 % yield by reaction with DIBAL in toluene at 95°C overnight. Treatment of (20a) with chloromethyl methyl ether and N,N-diisopropylethylamine at 65 ⁰ C overnight afforded the 3-methoxymethoxy ether (21a) in 77 % yield. Introduction of the 2-methoxy-group was carried out following the procedure of Paaren et al, US Patent No. 6,488,419 whereby metallation of compound (21a) was achieved by reaction with n- BuLi in THF at -65 ⁰ C. Subsequent addition of trimethyl borate and sodium perborate tetrahydrate gave the 2-hydroxy compound (22a) in 31 % yield. Methylation of compound (22a) with methyl iodide in DMF in the presence of potassium carbonate and tetra-n-butyl ammonium iodide at 45°C for four hours afforded the 2-methoxy derivative (23a) in quantitative yield. Hydrolysis of compound (23a) in THF with 6M HCl at room temperature for 48 hours followed by crystallization from ether gave compound (24a) in 87 % yield.

[00091] The syntheses of compounds (25a-30a) are outlined in Fig. 13. The 3,17β- dihydroxy-δ derivative (25a) was prepared as previously described (Rao et al., Steroids 2002;67: 1079-1089). This material was selectively acetylated with acetic anhydride in isopropanol in the presence of 2M NaOH to give the 3-acetate compound (26a) in 96 % yield. Treatment of (26a) with Jones reagent in acetone at 0 °C afforded the 17-ketone derivative (27a) in quantitative yield. The 17-ketone (27a) was converted to the 17- thioketal (28a) in 71 % yield by reaction with ethanedithiol and boron trifluoride diethyl etherate in acetic acid at room temperature for ninety minutes. This material was then refluxed with deactivated Raney nickel in acetone to give the δ ¹⁴ ' ¹⁶ derivative (29a) in 9.3 % yield. Simple base hydrolysis of (29a) with potassium carbonate in methanol/water gave the 3-hydroxy compound (30a) in quantitative yield.

[00092] The syntheses of the 2,8-dihydroxyhexahydrochrysene derivatives (36a and 40a) are outlined in Fig. 14. The 18-nor-13(17)-ene derivative (15a) was treated with acetic anhydride in pyridine at room temperature overnight to give the 3-acetate derivative (31a) in quantitative yield. Oxidative cleavage of the 13-17 double bond was carried out following the procedure of Yu et al (Org. Lett. 2004;6:3217-3219) using a catalytic amount of OsO ₄ in dioxane/water/t-BuOH in the presence of excess sodium periodate and pyridine to give the diketone derivative (32a) in 41% yield. Robinson

annulation of diketone (32a) was accomplished by refluxing in methanol with 10% KOH to give compound (33a) in quantitative yield. Compound (33a) was converted to the 3- acetate derivative (34a) in 51% yield by reaction with acetic anhydride in pyridine. Aromatization of compound (34a) to give the chrysene derivative (35a) was carried out in 70% yield following the procedure of Rao et al (Steroids 1994;59:621-627) using copper II bromide in acetonitrile. Hydrolysis of compound (35a) in methanol/water with potassium carbonate gave the 2,8-diol derivative (36a) in 88% yield. The diol (36a) was converted to the dimethoxymethyl ether (37a) in 88% yield by reaction with chloromethyl methyl ether and N,N-diisopropylethylamine in THF at 65 ⁰ C. Metallation of compound (37a) by reaction with sec-BuLi in THF at -65 ⁰ C followed by the addition of trimethyl borate and sodium perborate gave the 8-hydroxy compound (38a) in 84% yield. O-Methylation of compound (38a) with methyl iodide in DMF in the presence of potassium carbonate and tetra-n-butyl ammonium iodide afforded the 3,9- dimethoxychrysine derivative (39a) in 73% yield. Finally, hydrolysis of compound (39a) in THF with 6M HCl followed by subsequent chromatographic purification and crystallization gave the 3,9-dimethoxyhexahydrochrysene-2,8-diol derivative (40a) in 40% yield. Biological Activity

[00093] Suitable cell lines for testing biological activity include human umbilical vein endothelial cells (HUVEC), human breast carcinoma cells (MDA-MB-231) and human gliomablastoma cells (U87-MG).

[00094] Cell Cultures

[00095] Human umbilical vein endothelial cells (HUVEC) can be obtained from Clonetics

(San Diego, CA), MDA-MB-231, and U87-MG cells from ATCC (American Type Culture

Collection, Manassas, VA). HUVEC cultures are maintained for up to 5 passages in EGM

(Endothelial Growth Medium) containing bovine brain extract (Clonetics) and IX antibiotic- antimycotic (BioWhittaker, Walkersville, MD). MDA-MB-231 and U87-MG cells are maintained in DMEM/F12 (1:1) containing 10 % (v/v) fetal bovine serum (Hyclone

Laboratories, Logan, UT) and IX antibiotic-antimycotic.

[00096] In Vitro Inhibition o/Proliferation

[00097] Proliferation assays are performed by evaluating detection of DNA synthesis by the use of the 5-bromo-2'-deoxyuridine (BrdU) cell proliferation colorimetric ELISA kit from

Roche (Indianapolis, IN) according to the manufacturer's instructions. The cells are seeded

at 1,000 cells/well (MDA-MB-231 and U87-MG cells, anti-tumor activity) or 3,000 cells/well (HUVEC, anti-angiogenic activity) in a 96 well plate, allowed to attach overnight and then exposed to the compound to be tested for 48 hours.

[00098] The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

Example 1

3, 17β-Dihydroxy-18a-homoestra-\,3,5(10)-triene (2)

[00099] The 3 -hydroxy- 17 ketone derivative (1) was prepared from Norgestrel according to Rao et al., Steroids 1994;59:621-627. Under nitrogen, the 3-phenol (1, 9.4 g, 33 mmol) was dissolved in 50 ml of 1 :1 EtOHZH ₂ O. Sodium borohydride, (2.5 g, 66 mmol) was dissolved in 450 ml of 1 :1 EtOH/H ₂ O. The sodium borohydride solution was added to the steroid solution dropwise over 2 hours and stirred overnight. Analysis by TLC confirmed complete reaction (5% acetone/CH ₂ Cl ₂ ). Excess sodium borohydride was decomposed with acetic acid, the solvent was evaporated, and the residue extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, dried over sodium sulfate, filtered and evaporated in vacuo. The residue was crystallized from methanol/H ₂ O to give the pure 3,17-diol (2, 9.3 Ig, 98%) as a white powder, mp = 183 °C; FTIR (ATR) v _irax : 3377, 3270, 2930, 1614, and 1582 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.92 (t, J=7.4 Hz, 18a), 3.73(t,J=7.1 Hz, 17-H), 6.65 (t,J= 3.1 Hz, 4-H), 6.63 (dd, J, = 8.5 Hz, J ₂ = 2.6 Hz, 2-H), 7.15 (d, J= 8.7 Hz, 1-H). Anal. Calcd. for C ₎₉ H ₂₆ O ₂ -2/3MeOH: C, 76.77; H, 9.38. Found: C, 76.77; H, 9.35.

Example 2

18a-Homoestra-l, 3,5(10)-trien-3,17β-diyl-diacetate (3)

[OOOlOOJUnder nitrogen, the 3,17-diol (2, 9.41 g, 33 mmol) was dissolved in 100 ml of pyridine. Acetic anhydride, (11.5 ml, 0.12 mol) was added and the mixture stirred over the weekend. Analysis by TLC confirmed complete reaction (5% acetone/CH2Cl ₂ ). The reaction was quenched with methanol and the solvent removed in vacuo. The residue was crystallized from methanol to give the pure 3, 17-di acetate (3, 9.7g, 80%) as a white

powder, mp = 124-126 °C; FTIR (ATR) v _max : 2930, 1758, 1727, 1604, and 1576 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.96 (t, J= 7.6 Hz, 18a), 2.05 (s, 17-OAc), 2.78 (s, 3-OAc), 4.75 (t, J= 8.5 Hz, 17-H), 6.79 (t, J= 2.6 Hz, 4-H), 6.83 (dd, Ji, = 5.7 Hz, J ₂ = 2.7 Hz, 2-H), 7.27 (d, J = 5.7 Hz, 1-H). Anal. Calcd. for C ₂₃ H ₃₀ O ₄ : C, 74.56; H, 8.16. Found: C, 74.63; H, 8.11. Example 3

2-Acetyl-3-hydroxy-18a-homoestra-l,3,5(10)-trien-17β-yl-ace tate (4) fOOOlOlJUnder nitrogen, the 3,17-diacetate (3, 8.91 g, 24 mmol) was dissolved in 650 ml of CH ₂ Cl ₂ . Zirconium tetrachloride, (26 g, 0.11 mol) was added and the mixture was stirred 48 hours at room temperature. Analysis by TLC confirmed complete reaction (5% acetone/CH ₂ Cl ₂ ). The mixture was chilled to 0° C, quenched with 300 ml of water, and extracted with CH ₂ Cl ₂ (3x). The organic fractions were washed with water (3x) and brine, combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was crystallized from methanol to give the pure 2-acetyl compound (4, 5.18 g, 63%) as a white powder, mp = 240-242 ⁰ C; FTIR (ATR) v _max : 2943, 1714, 1636, 1614, and 1573 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.96 (t, J= 7.4 Hz, 18a), 2.05 (s, 17- OAc), 2.62 (s, 3-Ac), 4.76 (t, J=8.4 Hz, 17-H), 6.69 (s, 4-H), 7.59 (s, 1-H), 12.05 (s, 3- OH). Anal. Calcd. for C ₂₃ H ₃₀ O ₄ : C, 74.56; H, 8.16. Found: C, 74.36; H, 8.15.

Example 4

2-Acetyl-3-benzyloxy-18a-homoestra-l,3,5(10)-trien-17β-y l-acetate (5)

[000102]\Jndeτ nitrogen, the 2-acetyl compound (4, 5.18 g, 14 mmol) was dissolved in 165 ml of DMF. Potassium carbonate, (16.2 g, 0.12 mol) and benzyl chloride, (7.17 ml, 87 mmol) were added and the mixture stirred at 60° C for 48 hours. Analysis by TLC (5% Acetone/CH ₂ C1 ₂ ) confirmed a complete reaction. The reaction was quenched with 500 ml of water whereby a white flocky precipitate formed. The precipitate was collected by filtration, washed with water until the filtrate was neutral, and dried in vacuo. This material was crystallized from methanol/CH ₂ Cl ₂ to give the pure 3-benzyl ether (5, 5.59 g, 87%) as a white powder, mp = 213-215 ⁰ C; FTIR (ATR) v _max : 2939, 2869, 1734, 1657, and 1603 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.93 (t, J= 7.4 Hz, 18a), 2.05 (s, 17-OAc), 2.57 (s, 2- Ac), 4.74 (t, J= 8.3 Hz, 17-H), 5.13 (s, benzyl CH ₂ ), 6.73 (s, 4-H), 7.70 (s, 1-H), 7.34-7.45 (m, benzyl aromatic). Anal. Calcd. for C ₃₀ H ₃₆ O ₄ -l/6CH ₃ OH: C, 77.76; H, 7.93. Found: C, 77.89; H, 7.72.

Example 5

3-Benzyloxy-18a-homoestra-\,3,5(10)-trien-2, 17β-diyl-diacetate (6)

[000103]lJnder nitrogen, the 3-benzyl ether (5, 5.59 g, 12 mmol) was dissolved in 160 ml

Of CH ₂ Cl ₂ . Sodium phosphate (4.48 g, 32 mmol) and 3-chloroperoxybenzoic acid (4.17 g, 24 mmol) were added and the suspension stirred for 24 hours at room temperature. Water was added and the mixture extracted with CH ₂ Cl ₂ (3x). The organic fractions were washed with water (Ix), 10% sodium sulfite solution (Ix), and half saturated sodium bicarbonate solution (Ix). The organic fractions were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to give the pure 3-benzyl ether (6, 4.57 g, 79%) as a white foam, mp = 98-100 ⁰ C; FTIR (ATR) v _max : 2933, 1759, 1729, and 1612 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.92 (t, J= 7.4 Hz, 18a-H), 2.05 (s, 17-OAc), 2.25 (s, 2-OAc), 4.74 (t, J= 8.3 Hz, 17-H), 5.05 (s, benzyl CH ₂ ), 6.72 (s, 4-H), 6.95 (s, 1-H), 7.26-7.38 (m, benzyl aromatic). Anal. Calcd. for C ₃₀ H ₃₆ O ₅ : C, 75.60; H, 7.61. Found: C, 75.85; H, 7.69. Example 6

2, 17β-Dihydroxy-3-benzyloxy-l 8a-homoestra-\,3,5(l 0)-triene (7)

[000104]Unάer nitrogen, the 2,17-diacetate (6, 4.50 g, 9.5 mmol) was dissolved in 250 ml of methanol. To this solution 50 ml of IM NaOH solution was added and the mixture stirred at 60° C for 2 hours. Analysis by TLC (5% Acetone/CH ₂ C1 ₂ ) indicated an incomplete reaction. An additional 25 ml of IM NaOH solution was added and the mixture stirred an additional hour. Analysis by TLC at that time indicated a complete reaction. The reaction mixture was cooled to room temperature and glacial acetic acid was slowly added with stirring until the mixture was neutral. The mixture was diluted with 300 ml of water whereby a precipitate formed. Methanol was removed in vacuo and the residue was extracted with ethyl acetate (3x). The organic fractions were washed with water (3x) and brine, combined, dried over sodium sulfate, filtered and concentrated in vacuo. The solid residue was crystallized from ethyl acetate to give the pure 2,17-diol (7, 2.61 g, 71%) as a white powder, mp = 178 ⁰ C; FTIR (ATR) v _max : 3531, 3261, 2922, and 1604 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.98 (t, J= 7.55 Hz, 18a), 3.80 (t, J= 8.35 Hz, 17-H), 5.04 (s, benzyl CH ₂ ), 6.66 (s, 4-H), 6.92 (s, 1-H), 7.35- 7.45 (m, benzyl aromatic). Anal. Calcd. for C ₂₆ H ₃₂ O ₃ : C, 79.56; H, 8.22. Found: C, 79.38; H, 8.24.

Example 7

2-Methoxy-3-benzyloxy-18a-homoestra-\,3,5(10)-trien-l 7β-ol (8)

[000105] Under nitrogen, the 2,17-diol (7, 2.057 g, 5.2 mmol) was dissolved in 30 ml of

THF. Lithium hydroxide monohydrate (0.55 g, 1.3 mmol) and dimethyl sulfate (0.55 ml, 7.7mmol) were added and the mixture was heated to 60 ⁰ C overnight. Analysis by TLC (5% Acetone/CH2C1 ₂ ) confirmed complete a reaction and the mixture was cooled to room temperature. The solvent was removed in vacuo and the residue purified by flash chromatography (5% Acetone/CH ₂ Cl ₂ ). This material resisted crystallization and was obtained as a white foam which softened at 240-242 ⁰ C; FTIR (ATR) V _1113x : 3522, 2927, 2861, 1601, and 1506 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.99 (t, J= 7.4 Hz, 18a), 3.87 (s, 2- OCH ₃ ), 3.81 (t,J= 8.4 Hz, 17-H), 5.10 (s, benzyl CH ₂ ), 6.63 (s, 4-H), 6.48 (s, 1-H), 7.30-7.48 (m, benzyl aromatic). Anal. Calcd. for C ₂₇ H ₃₄ O ₃ -2/3H ₂ O C, 77.48; H, 8.51. Found: C, 77.92; H, 8.26.

Example 8

2-Methoxy-3, 17β-dihydroxy-l 8a-homoestra-\,3,5(J 0)-triene (9)

[000106] A mixture of the 2-methoxy compound (8, 200 mg, 0.5 mmol) and 5% palladium on carbon (200 mg) in ethanol (50ml) was hydrogenated in a Parr shaker apparatus at 40 psi hydrogen pressure overnight. Analysis by TLC (3% Acetone/CH ₂ d ₂ ) confirmed a complete reaction. The mixture was filtered through Celite while rinsing with dichloromethane. The solvent was removed in vacuo and the residue was crystallized from dichloromethane/hexanes to give the pure 3,17-diol (9, 126 mg, 81%) as a white powder, mp = 114-115 °C; FTIR (ATR) v _1Tiax : 3842, 3359, 2921, 2850, 1592, and 1505 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.00 (t, J =7.5 Hz, 18a), 3.86 (s, 2-OCH ₃ ), 3.77-3.821 (m, 17-H), 6.65 (s, 4-H), 6.79 (s, 1-H). Anal. Calcd. for C ₂ oH ₂₈ 0 ₃ -l/6Hexanes: C, 76.25; H, 9.24. Found: C, 76.20; H, 9.17.

Example 9

2, 17β-Dimethoxy-3-benzyloxy-18a-homoestra-\,3,5(10)-triene (10)

[000107]UndQV nitrogen, the 2-methoxy compound (8, 400 mg, 0.95 mmol) was dissolved in 20 ml of THF. This solution was added dropwise to 0.08 g (~2 mmol) of 60% NaH (60% disp. in mineral oil) in 10 ml of THF. The mixture was refluxed for 1 hour and cooled to room temperature. Methyl iodide (0.27 ml, 4.3 mmol) was added and the mixture stirred for 2 hours at room temperature. At this point TLC (5% Acetone/CH ₂ C12) showed no reaction. Additional methyl iodide (6 ml, 9.6 mmol) was added and the mixture again stirred overnight at room temperature. Analysis by TLC confirmed a complete reaction. The reaction was quenched with water and extracted with

ether (3x). The organic fractions were washed with water (3x) and brine, combined, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was crystallized from acetone/hexanes to give the pure 2,17-dimethoxy-3-benzyl ether (10, 266 mg, 64%). mp = 1 13-114 ⁰ C; FTIR (ATR) v _max : 2922, 2860, 1604, and 1514 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.98 (t, J = 7.5 Hz, 18a), 3.37 (s, 17- OCH ₃ ), 3.87 (s, 2-OCH ₃ ), 3.35 (t, J= 7.75 Hz, 17-H), 5.11 (s, benzyl CH ₂ ), 6.62 (s, 4-H), 6.85 (s, 1-H), 7.27-7.46 (m, benzyl aromatic). Anal. Calcd. for C ₂₈ H ₃₆ O ₃ : C, 79.96; H, 8.63. Found: C, 80.03; H, 8.68. Example 10

2, 17β-Dimethoxy-3-hydroxy-18a-homoestra-\,3,5(10)-triene (11)

[000108] A mixture of the 2, 17-dimethoxy-3 -benzyl ether (10, 150 mg, 0.35 mmol) and

5% palladium on carbon (150 mg) in ethanol (60ml) was hydrogenated in a Parr shaker apparatus at 40 psi hydrogen pressure overnight. Analysis by TLC (3% Acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The mixture was filtered through Celite while rinsing with dichloromethane. The solvent was removed in vacuo and the reside purified by flash chromatography (3% Acentone/CH ₂ Cl ₂ ). This material was triturated with cold ether, rinsed with pentane, and dried in vacuo to yield the pure 2,17-dimethoxy compound (11, 110 mg, 94%). mp = 161-162 °C; FTIR (ATR) v _inax : 3392, 2933, 2868, 1591, and 1506 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.97 (t, J= 7.4 Hz, 18a), 3.37 (s, 17- OCH ₃ ), 3.85 (s, 2-OCH ₃ ), 3.49 (t, J= 7.55 Hz, 17-H), 6.65 (s, 4-H), 6.80 (s, 1-H). Anal. Calcd. for C ₂₁ H ₃₀ O ₃ -1/6H ₂ O: C, 75.64; H, 9.17. Found: C, 75.54; H, 8.91. Example 11

3, 17β-bis(Methoxymethoxy)-18a-homoestra-l, 3, 5(10)-triene (12)

1000109] Under nitrogen, the diol (2, 47.2 g, 0.165 mol) was dissolved in 1.2 L of dry THF.

Chloromethyl methyl ether (62 ml, 0.816 mol) and diisopropylethylamine (170 ml, 0.976 mol) were added and the reaction heated to 65 ⁰ C overnight. The reaction was cooled to room temperature, quenched with 20% NH ₄ Cl, and extracted with EtOAc (3x). The organic phase washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the pure 3,17β-dimethoxymethyl ether (12, 48.2 g, 78 %) as a pale yellow oil. The material was used without further purification. FTIR (ATR) v _max : 2929, 2876, 1608, and 1496 cm ^"1 . NMR (300 MHz CDCl ₃ ), δ _H (ppm): 0.98 (t, J= 7.5 Hz, 18a), 3.38 (s, 17-OCH ₂ OCH ₃ ), 3.48 (s, 3-OCH ₂ OCH ₃ ), 4.64 (dd, J ₁ , = 9.0 Hz, J ₂ = 6.45 Hz, 17-OCH ₂ OCH ₃ ), 5.15 (s, 3-OCH ₂ OCH ₃ ), 6.78 (d, J= 2.4 Hz, 4-H), 6.84 (dd, J ₁ = 8.4

Hz, J ₂ = 2.4 Hz, 2-H), 7.21 (d ,J= 8.7 Hz, 1-H). Anal. Calcd. forC ₂₃ H ₃₄ O ₄ : C, 73.76; H, 9.15. Found: C, 73.34; H, 8.86. Example 12

2-Hydroxy-3,17β-bis(methoxymethoxy)-18a-homoestra-l,3,5(10) -triene (13) fOOOll OJ Under nitrogen, the dimethoxymethyl ether (12, 56.4 g, 0.151 mol) was dissolved in 1 L of dry THF and chilled to -78 ⁰ C. To this was added sec-BuLi (1.4M/cyclohexane, 215 ml, 0.3 mol) at such a rate the temperature did not exceed -65 ⁰ C and the reaction mixture stirred at -78 ⁰ C for 3 hours. Trimethyl borate (68 ml, 0.61 mol) was then added maintaining the temperature below -65 ⁰ C and the reaction stirred for 15 minutes at -78 °C and then warmed to 0 °C. The reaction was quenched with 1 L of 20% NH ₄ Cl solution, then allowed to come to room temperature and stirred for 1 hour. Sodium perborate tetrahydrate (93 g, 0.6 mol) was then added at such a rate that the temperature did not exceed 35 ⁰ C and the reaction carried out at room temperature overnight. The reaction mixture was concentrated in vacuo and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography (2% Acetone/CH ₂ C1 ₂ ) to give (13, 60.4 g, 84 %) as a yellow oil. FTIR (ATR) Vn _81x : 3407, 2930, 1589, and 1504 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.99 (t, J= 7.5 Hz, 18a), 3.36 (s, 17-OCH ₂ OCH ₃ ), 3.52 (s, 3-OCH ₂ OCH ₃ ), 4.63 (dd, Ji = 9.0 Hz, J ₂ = 6.6 Hz, 17-OCH ₂ OCH ₃ ), 5.16 (s, 3-OCH ₂ OCH ₃ ), 6.80 (s, 4-H), 6.90 (s, 1-H). This material was used without further purification.

Example 13

2-Methoxy-3, 17β-bis(methoxymethoxy)-18a-homoestra-l,3,5(10)-triene (14) [OOOlllJUnder nitrogen, the 2-hydroxy compound (13, 44.7 g, 0.115 mol) was dissolved in 500 ml of dry THF. Lithium hydroxide monohydrate (6 g, 0.143 mol) and dimethyl sulfate (12.2 ml, 0.128 mol) were added and the reaction was refluxed for 4 hours. Analysis by TLC (CH ₂ CL ₂ ) indicated a complete reaction. The reaction was cooled to room temperature, diluted with water, concentrated in vacuo, and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, filtered, dried over Na ₂ SO4, and concentrated in vacuo to give the 2-methoxy derivative (14, 53.7 g, >100 %) as a yellow oil which still contained some solvent. FTIR (ATR) v _ITiax : 2930, 1608, and 1507 cm ^" '. NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.98 (t, J= 7.5 Hz, 18a), 3.37 (s, 17-OCH ₂ OCH ₃ ), 3.51 (s, 3-OCH ₂ OCH ₃ ), 3.86 (s, 2-OCH ₃ ), 4.63 (dd, J ₁ = 9.0 Hz, J ₂ = 6.3 Hz, 17-OCH ₂ OCH ₃ ), 5.19

(s, 3-OCH ₂ OCH ₃ ), 6.84 (s, 4-H), 6.86 (s, 1-H). This material was used without further purification.

Example 14

2-Methoxy-3,17β-dihydroxy-18a-homoestra-l, 3,5(10)-triene (9)

[000112]Under nitrogen, 2-methoxy compound (14, 53.7 g, 0.132 mol) was hydrolyzed with a mixture of THF (300 ml) and 6M HCl (500 ml) at room temperature overnight. Analysis by TLC (2% Acetone/CH ₂ C1 ₂ ) indicated a complete reaction. The reaction was diluted with water, concentrated in vacuo and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by flash column (6% Acetone/CH ₂ C1 ₂ ). This material was crystallized from dichloromethane/hexanes to give the pure 3,17-diol (9, 30.3 g, 72 %) as yellow solid, mp = 114-115 °C; FTIR (ATR) v _max : 3484, 3362, 2921, 2872, and 1706 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.00 ft, J= 7.5 Hz, 18a), 3.84 (s, 2-OCH ₃ ), 6.64 (s, 4-H), 6.79 (s, 1-H). Anal. Calcd.for C ₂₀ H ₂₈ O ₃ l/6Hexanes: C, 76.25; H, 9.24. Found: C, 76.20; H, 9.17. Example 15

2-Methoxy-3-acetoxy-18a-homoestra-l,3,5(10)-trien-l 7β-ol (15) f000113JOnder nitrogen, the diol (9, 20 g, 62.3 mmol) was selectively acetylated in isopropanol with acetic anhydride (18 ml, 0.190 mmol) in the presence of NaOH (2M, 95 ml, 190 mmol) at room temperature for 90 minutes. Analysis by TLC (5% Acetone/CH ₂ C1 ₂ ) indicated a complete reaction. The reaction was slowly quenched with methanol, diluted with water, concentrated in vacuo and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the pure 3-acetate (15, 20.56 g, 91 %) as a yellow foam which resisted crystallization, mp = 84 ⁰ C; FTIR (ATR) v _max : 3523, 2931, 2870, 1751, and 1644 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.93 (t, J= 7.5 Hz, 18a), 2.29 (s, 3-OAc), 3.80 (s, 2-OCH ₃ ), 6.73 (s, 4-H), 6.89 (s, 1-H). Anal. Calcd. for C ₂₂ H ₃₀ O ₄ -2/3H ₂ O: C, 71.32; H, 8.52. Found: C, 71.15; H, 8.16. Example 16

2-Methoxy-3-acetoxy-18a-homoestra-l,3,5(10)-trien-l 7 -one (16)

[000114JUnder nitrogen, the 17-hydroxy compound (15, 20 g, 55.8 mmol) was dissolved in 400 ml of acetone and chilled to 0 ⁰ C. Jones's Reagent was slowly added with stirring until the yellow-orange color persisted (~40 ml). The reaction was stirred an additional five minutes then slowly quenched with isopropanol. The solution was concentrated in vacuo,

diluted with water, and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the 17-ketone (16, 19 g, 95.5 %). This material was used without further purification, mp = 159 ⁰ C; FTIR (ATR) V _1113x : 2932, 1762, 1731, 1614, and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.81 (t, J= 7.5 Hz, 18a), 2.31 (s, 3-OAc), 3.81 (s, 2- OCH ₃ ), 6.76 (s, 4-H), 6.89 (s, 1-H). Anal. Calcd. for C ₂₂ H ₂₈ O ₄ -1/6H ₂ O: C, 73.51; H, 7.94. Found: C, 73.73; H, 7.84.

Example 17

17,17-Ethylenedioxy-2-methoxy-l 8a-homoestra-l,3, 5(10)-trien-3-yl-acetate (17) [OOOUSJUndeτ nitrogen, the 17-ketone (16, 19 g, 53 mmol) was dissolved in 300 ml of

CH ₂ Cl ₂ . To this solution were added triethylorthoformate (28.8 ml, 173 mmol), ethylene glycol (19 ml, 341 mmol) and /?-toluenesulfonic acid monohydrate (0.77 g, 4 mmol) and the reaction mixture stirred at room temperature overnight. Analysis by TLC (2% acetone/CH ₂ Cl ₂ ) indicated a partial reaction. Additional triethylorthoformate (35 ml, 210 mmol) and ethylene glycol (10 ml, 179 mmol) were added and the reaction refluxed for an additional three hours. The reaction was cooled to room temperature, quenched with saturated NaHCO ₃ solution and extracted with CH ₂ Cl ₂ (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give a dark yellow oil. Analysis showed an incomplete reaction. The material was re-reacted over the weekend using 60 ml of triethylorthoformate, 40 ml of ethylene glycol and 0.8 g of tosic acid. Subsequent purification and re-acetylation afforded the ketal (17, 20 g, 94 %) as a yellow foam which resisted crystallization, mp - 97-98 °C; FTIR (ATR) v _max : 2936, 2875, 1763, 1614, and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.99 (t, J= 7.4 Hz, 18a), 2.31 (s, 3- OAc), 3.82 (s, 2-OCH ₃ ), 3.90 - 3.95 (m, 17-ketal CH ₂ 's), 6.74 (s, 4-H), 6.90 (s, 1-H). Anal. Calcd. for C ₂₄ H ₃₂ O ₅ -3/4H ₂ O: C, 69.62; H, 8.16. Found: C, 69.51; H, 8.00.

Example 18

17,17-Ethylenedioxy-16a-bromo-2-methoxy-18a-homoestra-l,3 ,5(10)-trien-3-yl-acetate

(18)

/000/76/Under nitrogen, the 17-ketal (17, 20 g, 50 mmol) was dissolved in 400 ml of THF and chilled to -5 ⁰ C. Benyltrimethylammonium tribromide (97%, 22.5 g, 56 mmol) was added in 5 g portions over 1/2 hour. Once the addition was complete, the reaction mixture was stirred at -5 ⁰ C overnight. The reaction was quenched with cold, saturated NaHCO ₃ and extracted with EtOAc (3x). The organic fractions were washed with saturated NaHCO ₃ solution (2x), 10% sodium thiosulfate solution, cold water (2x), and brine, combined, dried over Na ₂ SO ₄ ,

filtered, and concentrated in vacuo. The residue was purified by flash column and subsequently crystallized from CH ₂ Cl ₂ /hexanes to give the pure 16α-bromo compound (12, 22.94 g, 95 %) mp = 194 ⁰ C; FTIR (ATR) v _max : 2936, 2878, 1763, 1614, and 1508 cm ^" '. NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.00 (t, J = 7.05 Hz, 18a), 2.31 (s, 3-OAc), 3.81 (s, 2-OCH ₃ ), 3.96 - 4.21 (m, 17-ketal CH ₂ ¹ S), 4.69 (dd, Ji = 10.4 Hz, J ₂ = 3.4 Hz, 16-H), 6.74 (s, 4-H), 6.90 (s, 1-H). Anal. Calcd. for C ₂₄ H ₃ ,0 ₅ Br: C, 60.13; H, 6.52; Br, 16.67. Found: C, 59.96; H, 6.35; Br, 16.85.

Example 19

17,17-Ethylenedioxy-2-methoxy-18a-homoestra-l,3,5(10),14- tetraen-3-ol (19) and 17,17-Ethylenedioxy-2-methoxy-18a-homoestra-l,3,5(10),15-tet raen-3-ol (20) [000117]Undeτ nitrogen, the 16-bromo compound (18, 19 g, 19.8 mmol) was dissolved in

100 ml of dry xylenes. This solution was added to freshly prepared potassium tert-butoxide (43 g, 383 mmol) in 500 ml of dry xylenes and refluxed overnight. The reaction was cooled to room temperature, quenched with water and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give a dark brown residue. This material was purified via flash chromatography (1.5% acetone/CH ₂ Cl ₂ ) to give a mixture of the double bond isomers (19 and 20, 6.92 g, 49%) as a dark brown solid. This material was used without further purification. Example 20

2-Methoxy-3-hydroxy-18a-homoestra-l,3,5(10)14-tetraen-l 7-one (21) and 2-Methoxy-3-hydroxy-18a-homoestra-l,3,5(10)15-tetraen-l 7-one (22) fOOO118J\Jnάeτ nitrogen, the mixture of the double bond isomers (19 and 20, 8 g, 22.4 mmol) was dissolved in 400 ml of 6:1 acetone / water. /j-Toluenesulfonic acid monohydrate (0.41 g, 2.16 mmol) was added and the reaction mixture stirred at room temperature for 2.5 hours. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) indicated the reaction was complete. The reaction mixture was evaporated to 1/3 volume and extracted with CH ₂ Cl ₂ (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by flash chromatography (2.5 % acetone/CH ₂ Cl ₂ ) to give a mixture of the δ ¹⁴ and δ ¹⁵ isomers (21 and 22, 2.7 g) as well as the pure δ ¹⁵ derivative (22, 0.46 g, 45 %). The δ ¹⁵ compound (22) was crystallized from ether / hexanes. mp = 163 ⁰ C; FTIR (ATR) v _max : 3236, 2959, 2929, 1686, 1609, and 1523 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.79 (t, J= 7.5 Hz, 18a), 3.86 (s, 2-OCH ₃ ), 6.06 (dd, Ji = 6.0 Hz, J ₂ = 3.3 Hz, 16-H), 5.72 (s, 3-OH), 6.66 (s, 4-H), 6.77 (s, 1-H), 7.81 (dd, J, = 6.0

Hz, J ₂ = 2.7 Hz, 15-H). Anal. Calcd. for C ₂₀ H ₂₄ O ₃ : C, 76.89; H, 7.74. Found: C, 76.36; H,

7.71.

Example 21

2-Methoxy-18a-homoestra-l,3,5(10), 14,16-pentaen-3-l 7β-diyl-diacetate (23) [000119]lJnάer nitrogen, a mixture of δ ¹⁴ and δ ¹⁵ compounds (21 and 22, 2.7 g, 8.64 mmol) was dissolved in acetic anhydride (51 ml, 540 mmol). Isopropenyl acetate (50 ml, 454 mmol) and p-Toluenesulfonic acid monohydrate (1.0 g, 5.3 mmol) were added and the reaction mixture was refluxed overnight. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was cooled to room temperature, quenched with water and stirred for one hour. The mixture was then concentrated in vacuo and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by flash chromatography (2.5 % acetone/CH2Cl2) and subsequently crystallized from ether/hexanes to give the pure pentaene derivative (23, 1.6 g, 46.6 %) as a white solid, mp = 133 ⁰ C; FTIR (ATR) v _max : 2933, 2865, 1763, and 1615 cm ^'1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.53 (t, J- 7.4 Hz, 18a), 2.23 (s, 17-OAc), 2.31 (s, 3-OAc), 3.81 (s, 2-OCH ₃ ), 5.93 (t, J= 2.0 Hz, 15-H), 6.22 (d, J=2.7 Hz, 16-H), 6.79 (s, 4-H), 6.89 (s, 1-H). Anal. Calcd. for C ₂₄ H ₂₈ O ₅ : C, 72.71; H, 7.12. Found: C, 72.64; H, 7.14.

Example 22

2-Methoxy-3,l 7 β-dihydroxy-18a-homoestra-l, 3,5(10), 14-tetraene (24)

[000120]lJnder nitrogen, a solution of the dienol acetate (23, 1.5 g, 3.8 mmol) in ethanol (20 ml) and THF (20 ml) was chilled to 0 °C in an ice bath. An ice cold solution of sodium borohydride (0.9 g, 23.8 mmol) in ethanol/water (1:1, 50 ml) was added to the steroid solution. The reaction mixture was stirred for 1 hour, allowed to warm to room temperature, and then stirred overnight. Analysis by TLC (2 % acetone/CH ₂ Cl2) indicated a complete reaction. The reaction was quenched with acetic acid, concentrated to a small volume, and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by flash column to give the pure diol (24, 0.93 g, 69 %) as an amorphous powder, mp = 65 °C; FTIR (ATR) v _max : 3409, 2920, 2853, 1592, and 1506 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.89 (t, J= 7.5 Hz, 18a), 3.87 (s, 2- OCH ₃ ), 4.21 (t, J= 8.4 Hz, 17-H), 5.21 (m, 15-H), 6.67 (s, 4-H), 6.80 (s, 1-H). Anal. Calcd. for C ₂₀ H ₂₆ O ₃ l/2H ₂ O: C, 74.27; H, 8.41. Found: C, 74.33; H, 8.21.

Example 23

2-Methoxy-3-acetoxy-18a-homoestra-l,3,5(10), 14-tetraen-l 7β-ol (25)

[000121]XJnder nitrogen, the diol (24, 0.7 g, 2.2 mmol) was dissolved in 20 ml of isopropanol. Acetic anhydride (5 ml, 52 mmol) and 2M NaOH (20 ml, 40 mmol) were added and the reaction stirred at room temperature for 2 hours. Analysis by TLC (2 % acetone/CH2Cl ₂ ) indicated a complete reaction. The reaction was quenched with methanol, concentrated to a small volume and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , and concentrated in vacuo. Crystallization from ether / hexanes afforded the pure 3 -acetate (25, 0.61 g, 77 %) as a white solid, mp - 139 ⁰ C; FTIR (ATR) v _ma x: 3510, 2922, 2854, 1755, 1615, and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.88 (t, J = 7.4 Hz, 18a), 2.31 (s, 3-OAc), 3.80 (s, 2-OCH ₃ ), 4.19 (t, J= 8.6 Hz, 17-H), 5.34 (m, 15-H), 6.76 (s, 4-H), 6.90 (s, 1-H). Anal. Calcd. for C ₂₂ H ₂₈ O ₄ -I^H ₂ O: C, 72.30; H, 8.00. Found: C, 72.47; H, 7.70.

Example 24

2, 11§-Dimethoxy- 18a-homoestra-l, 3, 5(10), 14-tetraen-3-yl-acetate (26)

[000122JUnder nitrogen, fluoroboric acid (48 %, 2.5 ml, 18.9 mmol) was added to a solution of the 3-acetate (25, 0.5 g, 1.4 mmol) in CH ₂ Cl ₂ (20 ml) chilled to 0 °C, and stirred for 10 minutes. Trimethylsilyl diazomethane (2M/hexane, 1 ml, 2 mmol) was slowly added and the reaction mixture stirred for 20 minutes. An additional trimethylsilyl diazomethane solution (~lml) was added every 20 minutes. After the final addition, the reaction was stirred for an additional 1/2 hour. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was quenched with water and extracted with CH ₂ Cl ₂ (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash column (5 % acetone/CH ₂ Cl ₂ ) afforded the pure 17-methoxy derivative (26, 0.37 g, 71 %) as a yellow foam. This material was used in the subsequent reaction without further purification. FTIR (ATR) v _1Tiax : 2934, 2859, 1763, 1613, and 1508 cm ^" '. NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.85 (t, J= 7.5 Hz, 18a), 2.30 (s, 2-OAc), 3.41, (s, 17- OMe), 3.74 (t, J= 8.25 Hz, 17-H), 3.80 (s, 2-OCH ₃ ), 5.34 (m, 15-H), 6.75 (s, 4-H), 6.89 (s, 1- H). Example 25

2, 17β-Dimethoxy-3-hydroxy-18a-homoestra-l,3,5(10) 14-tetraene (27)

[000123JUnUQr nitrogen, the 3-acetate (26, 0.3 g, 0.81 mmol) was dissolved in 60 ml of

3:1 MeOH/H ₂ O. Potassium carbonate (0.44 g, 3.2 mmol) was added and the reaction mixture stirred at room temperature overnight. Analysis by TLC (5 % acetone/CH ₂ Cl2)

indicated a complete reaction. The reaction was quenched with water, concentrated, and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification of the residue by flash chromatography and subsequent crystallization from ether/hexanes afforded the pure 3-hydroxy (27, 0.2 g, 75 %) as a white solid, mp = 202 ⁰ C; FTIR (ATR) Vn _13x : 3383, 2935, 2834, 1618 and 1507 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.85 (t, J= 1.2 Hz, 18a), 3.42, (s, 17-OMe), 3.85 (t, J= 8.25 Hz, 17-H), 3.86 (s, 2-OCH ₃ ), 5.34 (m, 15- H), 6.66 (s, 4-H), 6.79 (s, 1-H). Anal. Calcd. for C ₂ iH ₂₈ O ₃ -l/6Et ₂ O: C, 76.36; H, 8.77. Found: C, 76.14; H, 8.55. Example 26

2-Methoxy-3,17β-dihydroxy-18a-homoestra-l, 3,5(10), 15-tetraene (28)

[000124]υo a solution of the 17-ketone (22, 0.4 g, 1.28 mmol), in ether (380 ml), chilled to -

5 ⁰ C, was added solid LiAlH ₄ (1.0 g, 26.4 mmol) in small portions, and the reaction mixture stirred for 1 hour. Analysis by TLC (5 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was quenched with the dropwise addition of 5% H ₂ SO ₄ and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (5 % acetone/CH ₂ Cl ₂ ) and subsequent crystallization from methanol afforded the pure diol (28, 0.15 g, 37 %) as a white solid, mp = 164 °C; FTIR (ATR) v _irax : 3519, 3240, 2919, 1589, and 1506 cm- ¹ . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.91 (t, J= 7.5 Hz, 18a), 3.87 (s, 2-OCH ₃ ), 5.71 (m, 16-H), 5.98 (dd, J ₁ = 7.8 Hz, J ₂ = 1.8 Hz, 15-H), 6.67 (s, 4-H), 6.79 (s, 1-H). Anal. Calcd. for C ₂₀ H ₂₆ O ₃ -l/2H ₂ O: C, 74.27; H, 8.41. Found: C, 74.38; H, 8.15.

Example 27

2-Methoxy-3-acetoxy-18a-homoestra-l,3,5(10),15-tetraen-l 7β-ol (29)

[000125]\Jnder nitrogen, the 3-hydroxy compound (28, 0.25 g, 0.79 mmol) was dissolved in

20 ml of isopropanol. Acetic anhydride (5 ml, 52 mmol) and 2M NaOH solution (20 ml, 40 mmol) were added and the reaction mixture was stirred at room temperature for 3 hours. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was quenched with methanol, concentrated to a small volume, and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by flash column (2 % acetone/CH ₂ Cl ₂ ) to give the pure 3-acetate (29, 0.17 g, 61 %) as a white foam. This material was used without further purification. FTIR (ATR) v _max : 3427, 2923, 2854, 1751, 1614, and

1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.90 (t, J= 7.5 Hz, 18a), 2.30 (s, 3-0Ac), 3.81 (s, 2-OCH ₃ ), 5.71 (m, 16-H), 5.97 (m, 15-H), 6.75 (s, 4-H), 6.89 (s, 1-H).

Example 28

2, 17β-Dimethoxy-18a-homoestra-l, 3, 5(10), 15-tetraen-3-yl-acetate (30) fOOOl 2 όJUnder nitrogen, 48 % fluoroboric acid (1.2 ml, 9.1 mmol) was added to a solution of the 3-acetate (29, 0.175 g, 0.49 mmol) in CH ₂ Cl ₂ (10 ml) chilled to 0 ⁰ C, and stirred for 10 minutes. Trimethylsilyl diazomethane (2M/hexanes, 8 ml, 16 mmol) was added dropwise over 2.5 hours. Once the addition was complete, the reaction was stirred an additional 90 minutes. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was quenched with water and extracted with CH ₂ Cl ₂ (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash column (5 % acetone/CH ₂ Cl ₂ ) afforded the pure 17β-methoxy (30, 0.13 g, 71 %) as a white foam which melted at 142 ⁰ C; FTIR (ATR) v _1Tiax : 2921, 1764, 1614 and 1511 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.88 (t, J= 7.5 Hz, 18a), 2.30 (s, 3- OAc), 3.47, (s, 17-OCH ₃ ), 3.81 (s, 2-OCH ₃ ), 5.80 (m, 16-H), 5.95 (dd, J _x = 6.0 Hz, J ₂ = 2.1 Hz, 15-H), 6.75 (s, 4-H), 6.89 (s, 1-H). Anal. Calcd. for C ₂₃ H ₃₀ O ₄ : C, 74.56; H, 8.16. Found: C, 74.20; H, 8.30.

Example 29

2,17β-Dimethoxy-3-hydroxy-18a-homoestra-l,3,5(10),15-tet raene (31)

[000127]Ondeτ nitrogen, the 3-acetate derivative (30, 0.1 g, 0.27 mmol) was dissolved in 20 ml of 3:1 MeOH/H ₂ O. Potassium carbonate (1.0 g, 7.2 mmol) was added and the reaction mixture stirred at room temperature for 2 hours. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was quenched with water, and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification of the residue by flash chromatography and subsequent crystallization from CH ₂ Cl ₂ /hexanes gave the pure 3- hydroxy compound (31, 0.067 g, 76 %) as a white solid, mp =180 °C; FTIR (ATR) v _max : 3378, 2930, 1617, and 1504 cm- ¹ . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.87 (t, J = 7.5 Hz, 18a), 3.46, (s, 17-OCH ₃ ), 3.87 (s, 2-OCH ₃ ), 5.80 (m, 16-H), 5.96 (m, 15-H), 6.65 (s, 4-H), 6.78 (s, 1-H). Anal. Calcd. for C _2l H ₂₈ O ₃ l/10H ₂ O: C, 76.38; H, 8.61. Found: C, 76.19; H, 8.52. Example 30

2-Methoxy-3-hydroxy-18a-homoestra-l,3,5(10)-trien-l7-onep -toluenesulfonylhydrazone

(32) f000128JTo a solution of the ketone (16, 1.0 g, 3.1 mmol) in MeOH (20 ml) was added p- toluene sulfonhydrazide (0.72 g, 3.9 mmol) and the reaction mixture was refluxed overnight.

Analysis by TLC (2 % acetone/C^C^) indicated an incomplete reaction. Hydrochloric acid (con, 4 drops) was added and the reaction mixture refluxed for an additional 4 hours. Analysis by TLC at this time indicated an incomplete reaction. Additional HCl was added (4 drops) and the reflux continued an additional 2 hours, after which time analysis by TLC indicated approximately 90 % completion. The reaction was cooled to room temperature and stored in the refrigerator over the weekend. The reaction mixture was diluted with water and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the hydrazone (32, 1.65 g) as a green/brown solid. The material was used in the subsequent reaction without further purification. FTIR (ATR) v _max : 3519, 3225, 2928, 1597, and 1508 cm ^" \ NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.50 (t, J= 7.2 Hz, 18a), 2.44 (d, J = 3.9 Hz, benzyl CH ₃ ), 3.87 (s, 2-OCH ₃ ), 6.64 (s, 4-H), 6.78 (s, 1-H), 7.316 (d, J= 7.5 Hz, benzyl CH ₂ ¹ S), 7.86 (d, J= 7.8 Hz, benzyl CH ₂ 's). Example 31

2-Methoxy-3-hydroxy-18a-homoestra-l, 3,5(10),16-tetraene (33) f000129JOnder nitrogen, the 17-p-toluenesulfonylhydrazone (32, 1.0 g, 2.1 mmol) was dissolved in dry THF (130 ml) and chilled to 0 ⁰ C in an ice/salt bath. To this solution was added chilled butyl lithium solution (2.5M/hexanes, 3.3 ml, 8.25 mmol). Once the addition was complete, the reaction mixture was stirred at room temperature for 72 hours. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was chilled to 0 ⁰ C, quenched with 20 % NH ₄ Cl solution, concentrated to a small volume, and extracted with EtOAc (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification of the residue by flash chromatography gave the pure tetraene (33, 0.12O g, 19 %) as a viscous oil. FTIR (ATR) v _raax : 3549, 2923, 2853, 1591 and 1504 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.79 (t, J = 7.5 Hz, 18a), 3.85 (s, 2-OCH ₃ ), 5.82 (m, 16-H), 6.04 (m, 17-H), 6.64 (s, 4-H), 6.78 (s, 1-H). Anal. Calcd. for C ₂₀ H ₂₆ O ₂ -l/2H ₂ 0: C, 79.70; H, 8.81. Found: C, 79.78; H, 8.87. Example 32 2-Methoxy-3-acetoxy-l 7β-hydroxyestra-\ ,3 ,5(10)-triene (2a)

[000130JTo a solution of 2-methoxyestradiol (Ia, 20 g, 58 mmol) in isopropanol (350 ml) was added sodium hydroxide solution (2M, 100 ml, 400 mmol) and acetic anhydride (16 ml, 169 mmol). The reaction mixture was stirred at room temperature and monitored by TLC (5 % acetone/CH ₂ Cl ₂ ) which indicated a complete reaction after two hours. The

reaction was slowly quenched with methanol, diluted with water, and concentrated in vacuo. The residue was acidified with HCl (3 M) and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the 3-acetate derivative (2a, 24.4 g) as a white solid, mp = 148 - 149 °C; FTIR (ATR) v _max : 3459, 2926, 2859, 1758, and 1505 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.76 (s, 18-CH ₃ ), 2.30 (s. 3-0Ac), 3.80 (s, 2-OCH ₃ ), 3.71 (t, J = 8.5 Hz, 17-H), 6.73 (s, 4-H), 6.90 (s, 1-H). Anal. Calcd. for C ₂ iH ₂₈ O ₄ : C, 73.26; H, 8.19. Found: C, 73.13; H, 8.04. Example 33

2-Methoxy-3-acetoxyestra-\ , 3,5(10)-trien-l 7-one (3a)

[000131]XJnder nitrogen, the 17-hydroxy compound (2a, 10 g, 29 mmol) was dissolved in 20 ml of acetone and chilled to 0 °C. Jones reagent was slowly added with stirring until the yellow-orange color persisted (18 ml). The reaction was stirred an additional five minutes then slowly quenched with isopropanol. The solution was concentrated in vacuo, diluted with water, and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the 17-ketone (3a, 9.07 g, 91 %) as a white solid, mp = 152 ⁰ C; FTIR (ATR) v _max : 2939, 1761, 1733, 1616, and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.92 (s, 18-CH ₃ ), 2.31 (s. 3-OAc), 3.81 (s, 2-OCH ₃ ), 6.77 (s, 4-H), 6.90 (s, 1- H). Anal. Calcd. for C _2I H ₂₆ O ₄ : C, 73.66; H, 7.65. Found: C, 73.53; H, 7.67. Example 34

2-Methoxy-3-hydroxyestra-\,3,5(10)-trien-l 7 -one (4a) f000132J\Jndeτ nitrogen, a solution of the 3-acetate derivative (3a, 11.3 g, 33 mmol) in 1 :1 THF/H ₂ O (100 ml) was treated with NaOH (2 M, 75 ml, 150 mmol) at room temperature for 1 hour. Analysis by TLC (5 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was cooled to 0 °C, quenched with 3 M_HC1, concentrated in vacuo, and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give 2-methoxyestrone (4a, 1O g, 100 %) as a white solid, mp = 189 - 191 ⁰ C) ;FTIR (ATR) V _1118x : 3359, 2928, 1720, 1588, and 1503 cm ^"1 . NMR (300 MHz, CDCl ₃ ) δ _H (ppm): 0.93 (s, 18-CH ₃ ), 3.87 (s, 2-OCH ₃ ), 5.51 (s, 3-OH), 6.67 (s, 4-H), 6.80 (s, 1-H). Example 35

2-Methoxy-3-methoxymethoxyestra- ,3,5(10)-trien-17-one (5a)

/0001337 Under nitrogen, 2-methoxyestrone (4a, 9.2 g, 30 mmol) was dissolved in 60 ml of THF. N,N-Diisopropylethylamine (35 ml, 200 mmol) and chloromethyl methyl ether (12.5 ml, 160 mmol) were added and the reaction mixture stirred at 65 ⁰ C overnight. The reaction was cooled to room temperature, quenched with 20 % NH ₄ Cl solution, and extracted with ethyl acetate (3x). The organic fractions washed with water (3x), and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the pure methoxymethyl ether (5a, 9.39 g, 89 %) as a white solid, mp = 117 °C; FTIR (ATR) v _max : 2928, 2854, 1731, 1606 and 1509 cm- ¹ . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.93 (s, 18- CH ₃ ), 3.53 (s, 3-OCH ₂ OCH ₃ ) 3.87 (s, 2-OCH ₃ ), 5.21 (s, 3-OCH ₂ OCH ₃ ), 6.85 (s, 4-H), 6.90 (s, 1-H). Anal. Calcd. for C ₂ iH ₂₈ O ₄ : C, 73.23; H, 8.19. Found: C, 73.09; H, 8.16. Example 36

2-Methoxy-3-methoxymethoxy-l 7a-cyano-l 7β-trimethylsifyloxyestra-l,3,5(10)triene (6a)

[000134]υo a solution of the 3 -methoxymethyl ether (5a, 4.9 g, 14.3 mmol) in chloroform (40 ml) were added zinc iodide (20 mg, 0.063 mmol) and trimethylsilylcyanide (3.0 ml, 23.5 mmol) and the reaction mixture stirred at room temperature overnight. The reaction mixture was quenched with water, concentrated in vacuo, and extracted with ethyl acetate (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give 7.O g of residue. Analysis by ¹ H NMR showed an incomplete reaction. The residue was re-reacted for another 4 hours, quenched and extracted as above and purified by flash column (2 % acetone/CH ₂ Cl ₂ ) to give 2.51 g of material. Analysis by ¹ H NMR showed the loss of the 3 -methoxymethyl ether.

/000135/In this material, the 3-hydroxyl function was re-protected by reaction with chloromethyl methyl ether and N,N-diisoproplyethylamine in THF at 65 ⁰ C overnight. The reaction was cooled to room temperature, quenched with 20 % NH ₄ Cl solution, and extracted with ethyl acetate (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the pure 3-rnethoxymethoxy-17α-cyano product (6a, 2.78 g, 40 %) as a yellow foam, mp = 63 ⁰ C; FTIR (ATR) V _n18x : 2929, 1608, and 1507 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.20 (s, 17B-OSiMe ₃ ), 0.79 (s, 18-CH ₃ ), 3.45 (s, 3-OCH ₂ OCH ₃ ) 3.8 (s, 2-OCH ₃ ), 5.12 (s, 3-

OCH ₂ OCH ₃ ), 6.74 (s, 4-H), 6.76 (s, 1-H). Anal. Calcd. for C ₂₅ H ₃₇ NO ₄ Si ^• 2/5 H ₂ O: C, 66.60; H, 8.45; N, 3.11. Found: C, 66.56; H, 8.17; N, 3.24. Example 37

2-Methoxy-3-methoxymethoxy-l 7a-aminomethylestra-l,3,5(10)-trien-l 7β-ol (7a)

[000136]\Jndeτ nitrogen, the 17α-cyano compound (6a, 2.7 g, 6.26 mmol) in THF (40 ml) was slowly added to an ice cold solution of LiAlH ₄ (IM in THF, 12.5 ml, 12.5 mmol). After the addition was complete, the reaction was allowed to come to room temperature and stirred overnight. The reaction mixture was chilled to 0 °C, quenched with saturated sodium sulfate solution, then solid sodium sulfate. The resulting mixture was filtered through Celite and the solids washed (3x) with THF. The combined filtrate was concentrated in vacuo and the residue extracted with ethyl acetate (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by flash column to give the pure 17α-aminomethyl (7a, 1.72 g, 75 %) as a yellow solid, mp = 114 - 115 °C; FTIR (ATR) V _1118x : 3350, 2918, 1607, and 1513 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.8 (s, 18- CH ₃ ), 1.274 (d, J = 7.5 Hz, IH), 3.40 (s, 3-OCH ₂ OCH ₃ ) 3.75 (s, 2-OCH ₃ ), 5.08 (s, 3- OCH ₂ OCH ₃ ), 6.73 (s, 4-H), 6.75 (s, 1-H). Anal. Calcd. for C ₂₂ H ₃₃ NO ₄ : C, 70.37; H, 8.86. Found: C, 70.22; H, 8.82. Example 38

2-Methoxy-3-methoxymethoxy-D-homoestra-\,2>,5(10)-trie n-17a-one (8a) and 2- Methoxy-3-Methoxymethoxy-D-homoestra-\ ,3 ,5( 1 O)-trien-l 7 -one (9a)

[000137/Under Nitrogen, the 17α-aminomethyl compound (7a, 1.72 g, 4.58 mmol) was dissolved in 48 ml of 5:1 HOAc/H ₂ O and chilled to 0 °C. Sodium Nitrite (1.67 g, 24.2 mmol) in 7 ml of H ₂ O was added dropwise over ¹ A hour, the solution was then stirred an additional 3 hours at 0 °C, then warmed to room temperature and stirred overnight. Analysis by TLC (5 % i-prOH/CH ₂ Cl ₂ ) indicated a complete reaction. The solvent was removed and the residue extracted with ethyl acetate (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue mixture was separated and purified by flash column (4:1 ether/hexanes) to give the pure 17a- and 17-ketones (8a, 1.0 g, 61 % and 9a, 90 mg, 5.5 % respectively) as white solids, mp (8a) = 115 ⁰ C; FTIR (ATR) v _max : 2936, 2866,

1697, 1606 and 1505 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.12 (s, 18-CH ₃ ), 3.50 (s, 3-OCH ₂ OCH ₃ ) 3.85 (s, 2-OCH ₃ ), 5.18 (s, 3-OCH ₂ OCH ₃ ), 6.84 (s, 4-H), 6.86 (s, 1-H). Anal. Calcd. for C ₂₂ H ₃₀ O ₄ : C, 73.71; H, 8.44. Found: C, 73.06; H, 8.29. f000138Jmp (9a) = 104 - 105 ⁰ C; FTIR (ATR) v _max : 2935, 1707, 1608 and 1507 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.85 (s, 18-CH ₃ ), 3.53 (s, 3-OCH ₂ OCH ₃ ) 3.86 (s, 2- OCH ₃ ), 5.21 (s, 3-OCH ₂ OCH ₃ ), 6.86 (s, 4-H), 6.89 (s, 1-H). Anal. Calcd. for C ₂₂ H ₃₀ O ₄ : C, 73.71 ; H, 8.44. Found: C, 73.31; H, 8.28. Example 39

2-Methoxy-D-homoestra-\,3,5(10)-trien-3,17aβ-diol (10a) and 2-Methoxy-D-homoestra- 1,3,5(10)-trien-3, 17aa-diol (Ha)

[000139/Under nitrogen, the 17a-ketone (8a, 1.0 g, 2.79 mmol) was dissolved in 20 ml of THF and cooled to 0 °C. Lithium tri-tert-butoxyaluminum hydride solution (IM in THF, 5.6 ml, 5.6 mmol) was added dropwise and the solution stirred at room temperature for 2.5 hours. The reaction was quenched with ethyl acetate, concentrated in vacuo, diluted with water, acidified with IM HCl, and extracted with ethyl acetate (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give a residue that was a mixture of the 17aβ- and 17aα-alcohols. The residue was separated by flash chromatography (4:1 ether/hexanes) and subsequently hydrolyzed with 6M HCl at 60 ⁰ C. Trituration from ether/hexanes yielded the 17aβ-OH and 17aα-OH compounds, (10a, 300 mg, 32 %) and (Ha, 100 mg, 10 %) as white solids, mp (10a) = 138 °C; FTIR (ATR) v _max : 3503, 3384, 2927, 2860, 1589, and 1505 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.85 (s, 18-CH ₃ ), 3.28 (dd, J, = 10.95 Hz, J ₂ = 4.65 Hz, 17-H), 3.87 (s, 2-OCH ₃ ), 6.64 (s, 4-H), 6.80 (s, 1-H). Anal. Calcd. for C ₂₀ H ₂₈ O ₃ -1/4H ₂ O: C, 74.85; H, 8.95. Found: C, 74.73; H, 8.88. [OOOUOJrrφ (Ha) = 146 °C; FTIR (ATR) v _max : 3561, 3499, 3271, 2937, 2863, 1608, and 1524 cm ^'1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.87 (s, 18-CH ₃ ), 3.43 (m, 17-H), 3.87 (s, 2-OCH ₃ ), 6.64 (s, 4-H), 6.81 (s, 1-H). Anal. Calcd. for C ₂₀ H ₂₈ O ₃ : C, 75.91 ; H, 8.92. Found: C, 75.49; H, 8.82.

Example 40

2-Methoxy-D-homoestra-\,3,5(10)-trien-3, 17β-diol (12a)

[000141]The 17-ketone (9a, 90 mg, 0.251 mmol) in 10 ml of THF was reacted similarly with lithium tri-tert-butoxyaluminum hydride solution (IM in THF, 0.5 ml, 0.5 mmol). Purification by flash chromatography (4:1 ether/hexanes), subsequent hydrolysis with 6M HCl at 60 °C and trituration from ether/hexanes yielded the pure 17β-OH compound (12a, 36 mg, 43 %) as a white solid, mp = 207 ⁰ C; FTIR (ATR) v _max : 3544, 3466, 3272, 2925, 1595, 1513 and 1501 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.10 (s, 18-CH ₃ ), 3.86 (s, 2-OCH ₃ ), 4.15 (m, 17-H), 6.64 (s, 4-H), 6.80 (s, 1-H). Anal. Calcd. for C ₂₀ H ₂₈ O ₃ -2/5H ₂ O: C, 74.22; H, 8.97. Found: C, 74.02; H, 8.66. Example 41

2-Methoxy-3-methoxymethoxyestra-\,3,5(10)-trien-l 7β-ol (13a)

[000142]Under nitrogen, sodium borohydride (1.3 g, 34.35 mmol) in 1 :1 EtOH:H ₂ O (65 ml) was added dropwise to a solution of the 17-ketone (5a, 4.76 g, 13.74 mmol) in 1 :1 THF:EtOH (250 ml) and stirred at room temperature for Vi hour. Analysis by TLC (5 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was chilled in an ice bath, quenched with acetic acid until acidic, then concentrated in vacuo. The residue was diluted with water, acidified with 6M HCl, and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ ,filtered, and concentrated in vacuo. The residue was purified by flash chromatography (5 % acetone/CH ₂ Cl ₂ ) to give the 17-hydroxy compound (13a, 4.74g, 99 %) as a yellow semisolid that still contained some solvent. FTIR (ATR) v _max : 3422, 2923, 2866, 1608 and 1507 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.79 (s, 18-CH ₃ ), 3.51 (s, 3-OCH ₂ OCH ₃ ) 3.73 (t, J= 8.4 Hz, 17-H), 3.85 (s, 2-OCH ₃ ), 5.19 (s, 3-OCH ₂ OCH ₃ ), 6.84 (s, 4-H), 6.87 (s, 1-H). Anal. Calcd. for C ₂ iH ₃₀ O ₄ -l/6 H ₂ O: C, 72.18; H, 8.75. Found: C, 72.09; H, 8.52. Example 42

2-Methoxy-3-methoxymethoxyestra-\, 3,5(10)-trien-l 7β-yl-tosylate (14a) f000143J\Jndev nitrogen, />toluenesulfonic anhydride (97 %, 5.5 g, 16.3 mmol) was added to a solution of the 17-hydroxy compound (13a, 4.74 g, 13.7 mmol) in anhydrous pyridine (80 ml) at 0 ⁰ C and the mixture was stirred for 2 hours at room temperature.

Analysis by TLC (5 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was quenched with water and extracted with ethyl acetate (3x). The organic fractions were washed with cold 2M HCl, water, cold saturated NaHCO ₃ solution, and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash column (2 % acetone/CH ₂ Cl ₂ ) and crystallization from acetone/hexanes afforded the 17-tosylate (14a, 6.7 g, 95 %) as a white solid, mp = 115 ⁰ C; FTIR v _max (ATR) : 2969, 2922, 2868, 1598, and 1515 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.83 (s, 18-CH ₃ ), 2.46 (s, OTs-CH ₂ ), 3.50 (s, 3-OCH ₂ OCH ₃ ), 3.84 (s, 2-OCH ₃ ), 4.36 (t, J = 8.1 Hz, 17-H), 5.18 (s, 3-OCH ₂ OCH ₃ ), 6.79 (s, 4-H), 6.85 (s, 1-H), 7.34 (d, J = 3.9 Hz, OTs-ArH), 7.80 (d, J = 4.0 Hz, OTs-ArH). Anal. Calcd. for C ₂₈ H ₃₆ O ₆ S: C, 67.17; H, 7.25; 5, 6.40. Found: C, 67.02; H, 6.98; 5, 6.35. Example 43

2-Methoxy3-hydroxy-l 7-methylgona-\,3,5(10), 13(17)-tetraene (15a) f000144J\Jndev nitrogen, the 17-tosylate derivative (14a, 6.23 g, 12.5 mmol) in benzene (125 ml) was added dropwise to a solution of ethylmagnesium bromide (3M/ether, 53 ml, 159 mmol). The ether was distilled off, the solution was refluxed for 3 hours and then stirred overnight at room temperature. The reaction was cooled to 0 ⁰ C, quenched with the dropwise addition of cold 2M H ₂ SO ₄ , and extracted with ethyl acetate (3x). The organic fractions were washed with cold saturated NaHCO ₃ solution, water, and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash column (2 % acetone/CH ₂ Cl ₂ ) and crystallization from acetone/hexanes afforded the 17- methyl product (15a, 2.86 g, 79 %) as a white solid, mp = 108 ⁰ C; FTIR (ATR) v _max : 3444, 2911, 2855, 2832, 1591, and 1505 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.65 (s, 17-CH ₃ ), 3.87 (s, 2-OCH ₃ ), 5.42 (s, 3-OH), 6.64 (s, 4-H), 6.83 (s, 1-H). Anal. Calcd. for C ₁₉ H ₂₄ O ₂ : C, 80.24; H, 8.51. Found: C, 79.86; H, 8.14. Example 44

2-Methoxy-3-methoxymethoxy-21-trimethylsilyl-19-norpregna -\,3,5(10),16- tetraene-20-yne (16a)

[000145]\Jnder nitrogen, a solution of trimethylsilylacetylene (1 ml, 7 mmol) in THF (10 ml) was cooled to -78 °C. Butyl lithium (1.6 M in hexanes, 4.4 ml, 7 mmol) was slowly added and the mixture stirred at -78 ⁰ C for 10 minutes, and then allowed to warm to 0 ⁰ C

in an ice bath. After stirring at 0 ⁰ C for ¹ A hour, the mixture was cooled again to -78 ⁰ C and transferred to a solution of the ketone (5a, 1 g, 2.9 mmol) in 12 ml of 2:1 THF/benzene, also at -78 °C. The reaction mixture was stirred for 15 minutes then allowed to warm to room temperature. After stirring at room temperature for 2 hours, TLC (2 % acetone/CH ₂ Cl ₂ ) of a quenched aliquot indicated complete formation of the lithium intermediate. Methanesulfonyl chloride (1 ml, 4.77 mmol) was added followed ¹ A hour later by pyridine (2 ml). The reaction mixture was then stirred overnight at room temperature. The reaction mixture was quenched with saturated NaHCO ₃ solution and extracted with ethyl acetate (3x). The organic fractions were washed with water, IM HCl (2x), water and brine. The combined organic fractions were dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give 1.6 g of a dark violet residue. This material was passed through a short silica column (2 % acetone/CH ₂ Cl ₂ ) to give a mixture of the 3- methoxymethyl and 3-hydroxy derivatives. This mixture was used directly in the subsequent reaction. Example 45

2.4 2-Methoxy-l 7 -ethynylestra-X , 3,5(1 O), 16-tetraen-3-ol (17a)

[000146]\Jnder nitrogen, the 21-trimethylsilyl mixture (16a, 0.85 g, 2 mmol) was dissolved in 15 ml of THF. Tetrabutylammonium fluoride (1M/THF, 3.7 ml, 3.7 mmol) was added and the reaction stirred at room temperature for 2 hours. The reaction was quenched with water and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give 0.65 g of a brown foam. The residue was twice purified by flash column (1 % acetone/CH ₂ Cl ₂ ), then (7:3 CH ₂ Cl ₂ /hexanes) to give 120 mg of material. This material was twice crystallized from acetone/hexanes to give the pure product (17a, 40 mg, 6.4 %). mp = 202 ⁰ C; FTIR (ATR) v _max :3490, 3293, 2932, 1591, and 1504 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 0.90 (s, 18-CH ₃ ), 1.55 (s, 17-CCH), 3.87 (s, 2-OCH ₃ ), 6.16 (t, J = 2.6 Hz, 16-H), 6.66 (s, 4-H), 6.79 (s, 1-H). Anal. Calcd. for C ₂ ]H ₂₄ O ₂ : C, 81.78; H, 7.84. Found: C, 81.73; H, 7.97. Example 46

17,17a-Dimethyl-3-methoxy-D-homogona-\,3,5(10), 13, 15, 17(17a)-hexaene (19a)

[000147J Compound (19a) was prepared from mestranol (18a) by reacting with formic acid as reported in the literature [Vincze et al., Steroids 1993;58: 220-224].

Example 47

17, 17a-Dimethyl-3-hydroxy-D-homogona-\,3,5(10),13,15, 17(17a)-hexaene (20a) f000148J\Jnder nitrogen, the 3-methoxy derivative (19a, 1.0 g, 3.41 mmol) was dissolved in toluene (10 ml) and treated with DIBAL (lM/toluene, 17 ml, 17 mmol) at 95 ⁰ C overnight. Analysis by TLC (5 % acetone/CH ₂ Cl ₂ ) showed a complete reaction. The reaction mixture was quenched with 1 :1 ethanol/water, the resulting precipitate was dissolved with 5 % HCl and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash column (2 % acetone/CH ₂ Cl ₂ ) gave the 3 -hydroxy compound (20a, 0.73 g. 76 %) as a white solid, mp = 177 ⁰ C; FTIR (ATR) v _max : 3372, 2918, 2828, and 1607 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.19 (s, 17-CH ₃ ), 2.30 (s, Ha-CH ₃ ), 4.82, (s, 3-OH), 6.62 (d, J = 2.7 Hz, 4-H), 6.68 (dd, J ₁ = 8.1 Hz, J ₂ = 3.0 Hz, 2-H), 7.05 (d, J = 9.6 Hz, 1-H), 7.20 (d, J = 8.1 Hz, 15-H), 7.28 (d, J = 8.4 Hz, 16-H). Anal. Calcd. for C ₂₀ H ₂₂ O -1/10CH ₂ Cl ₂ : C, 84.15; H, 7.80. Found: C, 84.08; H, 7.86. Example 48

17,17a-Dimethyl-3-methoxymethoxy-D-homogona-\,3,5(10), 13,15,17(17a)-hexaene (21a)

[000149]XJnder nitrogen, the 3 -hydroxy compound (20a, 0.9 g, 3.23 mmol) was dissolved in 15 m of dry THF. Chloromethyl methyl ether (1.25 ml, 16.5 mmol) and diisoproplyethylamine (3.4 ml, 19.5 mmol) were added and the reaction mixture heated to 65 °C overnight. Analysis by TLC (2 % acetone/CH ₂ Cl ₂ ) showed a complete reaction. The reaction mixture was cooled to room temperature, quenched with 20 % NH ₄ Cl solution, and extracted with ethyl aceate (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ ,fϊltered, and concentrated in vacuo. Purification by flash column (1.5 % acetone/CH ₂ Cl ₂ ) gave the 3-methoxymethoxy ether product (21a, 0.8 g, 77 %) as a white solid, mp = 102 ⁰ C; FTIR (ATR) v _max : 2953, 2826, 1613 and 1575 cm ^'1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.16 (s, 17-CH ₃ ), 2.27 (s, 17α- CH ₃ ), 3.47 (s, 3-OCH ₂ OCH ₃ ), 5.18 (m, 3-OCH ₂ OCH ₃ ), 6.84 (d, J = 2.4 Hz, 4-H), 6.88 (dd, J, = 8.7 Hz, J ₂ = 3.0 Hz, 2-H), 7.02 (d, J = 8.1 Hz, 1-H), 7.18 (d, J= 8.4 Hz, 15-H), 7.30 (d, J = 8.4 Hz, 16-H). Anal. Calcd. for C ₂₂ H ₂₆ O ₂ ^• 1/4H ₂ O : C, 80.82; H, 8.17. Found: C, 80.96; H, 7.97.

Example 49

17,17a-Dimethyl-2-hydroxy-3-methoxymethoxy-D-homogona- 1 ,3,5(10), 13,15, 17(17a)-hexaene (22a)

[000150]Under nitrogen, the 3-methoxymethoxy ether (21a, 0.8 g, 2.48 mmol) was dissolved in 20 ml of dry THF and chilled to -78 °C. To this was added sec-BuLi (1.4 M/cyclohexane, 3.5 ml, 4.9 mmol) at such a rate that the temperature did not exceed -65 °C. The reaction mixture was stirred at -78 °C for 3 hours. Trimethyl borate (1.1 ml, 9.9 mmol) was then added at such a rate that the temperature did not exceed -65 ⁰ C and the mixture stirred for one hour at -78 ⁰ C. The reaction mixture was quenched with 15 ml of 20 % NH ₄ Cl solution, allowed to come to room temperature, and stirred for 1 hour. Sodium perborate tetrahydrate (1.5 g, 9.7 mmol) was then added at such a rate that the temperature did not exceed 35 °C and the reaction mixture was stirred at room temperature overnight. The reaction mixture was quenched with water and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by flash column (8:2 CH ₂ Cl ₂ /hexanes) to give the 2-hydroxy compound (22a, 0.26 g, 31 %) as a white solid, mp = 145 - 146 °C; FTIR (ATR) v _max : 3354, 2949, 1618, and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.16 (s, 17-CH ₃ ), 2.27 (s, 17a-CH ₃ ), 3.50 (s, 3- OCH ₂ OCH ₃ ), 5.15 (m, 3-OCH ₂ OCH ₃ ), 6.85 (s, 1-H), 7.02 (d, J = 7.8 Hz, 15-H), 7.17 (d, J = 8.4 Hz, 16-H). Anal. Calcd. for C ₂₂ H ₂₆ O ₃ -1/3H ₂ O: C, 76.73; H, 7.80. Found: C, 76.76; H, 7.59. Example 50

17,17a-Dimethyl-2-methoxy-3-methoxymethoxy-D-homogona- 1 ,3 ,5(10), 13, 15, 17(17a)-hexaene (23a)

[00015I]To a solution of the 2-hydroxy (22a, 0.46 g, 1.36 mmol) in DMF (20ml) was added potassium carbonate (1.8 g, 13 mmol) and tetrabutylammonium iodide (30 mg, 0.08 mmol) and the reaction mixture stirred for 15 minutes. Methyl iodide (1.9 ml, 30.5 mmol) was added and the mixture stirred at 45 ⁰ C for 4 hours then stirred overnight at room temperature Analysis by TLC (7:3 ether/hexanes) showed a complete reaction. The reaction mixture was quenched with water and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ ,

filtered, and concentrated in vacuo. Purification by flash column (7:3 ether/hexanes) gave the 2-methoxy derivative (23a, 0.49 g, >100 %) as a white foam, mp = 82 ⁰ C; FTIR (ATR) v _max : 2917, 2857, 1608 and 1514 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.17 (s, 17-CH ₃ ), 2.28 (s, Ha-CH ₃ ), 3.52 (s, 3-OCH ₂ OCH ₃ ), 3.89 (s, 2-OCH ₃ ), 5.21 (m, 3- OCH ₂ OCH ₃ ), 6.94 (s, 4-H), 6.95 (s, 1-H), 7.03 (d, J= 8.7 Hz, 15-H), 7.20 (d, J = 8.1 Hz, 16-H). Anal. Calcd. for C ₂₃ H ₂₈ O ₃ : C, 78.38; H, 8.01. Found: C, 78.07; H, 8.08. Example 51

17,17a-Dimethyl-2-methoxy-3-hydroxy-D-homogona-l ,3,5(10), 13,15, 17(17a)-hexaene (24a)

/000752/Under nitrogen, the 3-methoxymethoxy ether (23a, 0.46 g, 1.3 mmol) was hydrolyzed with 6M _. HCl (15 ml) in THF (20 ml) at room temperature over the weekend. Analysis by TLC (7:3 ether/hexanes) indicated a complete reaction. The reaction mixture was quenched with water and extracted with ethyl aceate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash column (1 % acetone/CH ₂ Cl ₂ ) and subsequent crystallization from ether gave the 3-hydroxy derivative (24a, 0.35 g, 87 %) as a white solid, mp = 177 ⁰ C; FTIR (ATR) v _max : 3511, 2920, 2841, 1592, and 1514 cm ^'1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.19 (s, 17-CH ₃ ), 2.30 (s, 17a-CH ₃ ), 3.91 (s, 2-OCH ₃ ), 5.48 (s, 3-OH), 6.72 (s, 4-H), 6.91 (s, 1-H), 7.05 (d, J = 8.1 Hz, 15-H), 7.22 (d, J = 7.8 Hz, 16-H). Anal. Calcd. for C _{2 I} H ₂₄ O ₂ : C, 81.78; H, 7.84. Found: C, 81.05; H, 7.75. Example 52

2-Methoxy-3-acetoxyestra-\,3,5(10), 14-tetraen-l 7β-ol (26a)

[000153]υo a solution of compound (25a) in isopropanol (20 ml) was added 2M sodium hydroxide (15 ml, 30 mmol) and acetic anhydride (0.7 ml, 7.4 mmol) and the reaction mixture stirred at room temperature for two hours. Analysis by TLC (5 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was slowly quenched with methanol, diluted with water, concentrated in vacuo and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the 3-acetate derivative (26a, 0.82 g, 96 %) as a white solid, mp = 84 ⁰ C; FTIR (ATR) v _max : 3425, 2927, 2843,1757, 1615, and 1508 cm ^" '. NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.03 (s, 18-CH ₃ ), 2.31 (s, 3-OAc), 3.81 (s, 2-

OMe), 4.11 (t, J= 8.2 Hz, 17-H), 5.22 (s, 15-H), 6.77 (s, 4-H), 6.93 (s, 1-H). Anal. Calcd. for C ₂ ,H ₂₆ O ₄ : C, 73.66; H, 7.65. Found: C, 73.77; H, 7.59. Example 53

2-Methoxy-3-acetoxyestra- 1,3, 5(10)J 4-tetraen-l 7 -one (27a) f000154J\Jnder nitrogen, the 17-hydroxy compound (26a, 0.82 g, 2.4 mmol) was dissolved in 20 ml of acetone and chilled to 0 ⁰ C. Jones reagent was slowly added with stirring until the yellow-orange color persisted (~3 ml). The reaction was stirred an additional five minutes then slowly quenched with isopropanol. The solution was concentrated in vacuo, diluted with water, and extracted with ethyl acetate (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the 17-ketone (27a, 0.82 g, 100 %) as a yellow solid, mp = 133 ⁰ C; FTIR (ATR) v _max : 2966, 2938, 1762, 1734, 1615 and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.18 (s, 18-CH ₃ ), 2.31 (s, 3-0Ac), 3.81 (s, 2-0Me), 5.63 (s, 15-H), 6.79 (s, 4-H), 6.89 (s, 1-H). Anal. Calcd. for C ₂₁ H ₂₄ O ₄ ^• 1/10 H ₂ O: C, 73.70; H, 7.13. Found: C, 73.81; H, 7.06. Example 54

2-Methoxy-3-acetoxyestra-\,3,5(10), 14-tetraene-l 7, 17-ethylenethioketal (28a)

[000155]υo a solution of the 17-ketone compound (27a, 0.8 g, 2.4 mmol) in acetic acid (35 ml) was added ethanedithiol (1.9 ml, 22.6 mmol) and boron trifluoride diethyl etherate (1.9 ml, 15.4 mmol) and the reaction mixture was stirred at room temperature for ninety minutes. Analysis by TLC (5 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction was quenched with water and extracted with ether (3x). The organic fractions were washed with saturated NaHCO ₃ solution (2x), water and brine. The combined organic fractions were dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (CH ₂ Cl ₂ ) gave the thioketal derivative (28a, 0.7 g, 71 %) as a white foam, mp = 123 ⁰ C; FTIR (ATR) V _018x : 2926, 2857, 2835, 1762, 1614, and 1508 cml NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.12 (s, 18-CH ₃ ), 2.31 (s, 3-OAc), 3.15-3.37 (m, 17-thioketal CH ₂ 's) 3.82 (s, 2-OMe), 5.47 (s, 15-H), 6.77 (s, 4-H), 6.94 (s, 1-H). Anal. Calcd. for C ₂₃ H ₂₈ O ₃ S ₂ : C, 66.31 ; H, 6.77; S, 15.39. Found: C, 66.33; H, 6.83; S, 15.62.

Example 55

2-Methoxyestra-\, 3,5(10), 14,16-pentaen-3-yl-acetate (29a) thioketal compound (28a, 0.68 g, 1.63 mmol) in acetone (20 ml) was added to a mixture of deactivated Raney nickel (5 g) in acetone (50 ml) and the mixture refluxed for eight hours. Analysis by TLC (3 % acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was filtered through Celite, and concentrated in vacuo. The residue was purified by flash chromatography (1 :1 ether/hexanes) to give the δ ¹⁴ ' ¹⁶ derivative (29a, 60 mg, 9.3 %) as a white solid, mp = 88 °C. FTIR (ATR) v _max : 2924, 2856, 1762, 1614 and 1509 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.08 (s, 18-CH ₃ ), 2.31 (s, 3-0Ac), 3.81 (s, 2-OMe), 5.92 (s, 15-H), 6.33 (dd, J ₁ = 4.8 Hz, J ₂ = 1.8 Hz, 16- H), 6.38 (d, J = 2.4 Hz, 17-H), 6.79 (s, 4-H), 6.92 (s, 1-H). Anal. Calcd. for C ₂₃ H ₂₈ O ₃ ^• 1/2 H ₂ O: C, 75.65; H, 7.56. Found: C, 75.60; H, 7.58. Example 56

2-Methoxy-3-hydroxyestra- \, 3,5(10), 14,16-pentaene (30a)

[000157]υo a solution of the 3-acetate compound (29a, 50 mg, 0.15 mmol) in 90 % methanol/water (20 ml) was added potassium carbonate (0.2 g, 1.45 mmol) and the reaction mixture stirred at room temperature for two hours. Analysis by TLC (1 :1 ether/hexanes) indicated a complete reaction. The reaction mixture was acidified with 0.5M HCl and extracted with ether (3x). The organic fractions were washed with water and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by flash chromatography (1 :1 ether/hexanes) to give the 3-hydroxy compound (30a, 44 mg, 100 %) as a clear oil. FTIR (ATR) v _max : 3549, 2923, 2854, 1592, and 1505 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.08 (s, 18-CH ₃ ), 3.87 (s, 2-OMe), 5.92 (s, 15-H), 6.33 (dd, J ₁ = 5.1 Hz, J ₂ = 1.8 Hz, 16-H), 6.37 (d, J = 2.4 Hz, 17-H), 6.70 (s, 4-H), 6.82 (s, 1-H). Anal. Calcd. for C ₉ H ₂₂ O ₂ ^• 1/4 H ₂ O: C, 79.55; H, 7.91. Found: C, 79.51 ; H, 7.92. Example 57

2-Methoxy-3-acetoxy-l 7 -methylgona-X ,3,5(10), 13(17)-tetraene (31a) f000158JVndev nitrogen, the 3-hydroxy compound (15a, 14.2 g, 50 mmol) was acetylated in pyridine (250 ml) with acetic anhydride (60 ml, 634.8 mmol) at room temperature overnight. Analysis by TLC (1% acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was quenched with methanol and concentrated in vacuo to

give the 3-OAc (3, 17.5 g, 100%) as a yellow solid. This material was used in the subsequent reaction without further purification, mp = 91 ⁰ C; FTIR (ATR) v _max : 2924, 2837, 1762, 1615, and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 1.6 (s, 17-CH ₃ ), 2.30 (s, 3-OAc), 3.80 (s, 2-OCH ₃ ), 6.72 (s, 4-H), 6.92 (s, 1-H). Anal. Calcd. for C ₂ ]H ₂₆ O ₃ : C, 77.27; H, 8.03. Found: C, 77.20; H, 7.96. Example 58

(8s, 4bS, 8aR)-3-Methoxy- 7-oxo-8-(3-oxobutyl)-5, 6, 8, 9, 10, 4b, 8a-heptahydrophenanthra-2- yl acetate (32a)

[000159] Under nitrogen, the 3-acetate derivative (31a, 2.0 g, 6.1 mmol) in 3:1 dioxane/water (10 ml) was treated with a solution of OsO ₄ (6.2 mg, 0.024 mmol) in t- BuOH (2 ml). Sodium periodate (0.52 g, 2.43 mmol) and pyridine (0.1 ml, 1.24 mmol) were added and the mixture was heated at 65-7O ⁰ C for 3 hours and then stirred at room temperature overnight. Analysis by TLC (CH ₂ Cl ₂ ) indicated an incomplete reaction. Additional OsO ₄ in t-BuOH (3 mg in 1 ml, 0.012 mmol) was added and the reaction heated at 65-70 ⁰ C for 4 hours. Analysis by TLC showed disappearance of the starting material. The reaction mixture was cooled to room temperature, quenched with water, and extracted with EtOAc (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (5% acetone/CH ₂ Cl ₂ ) gave compound (32a, 0.9 g, 41%) as a white solid, mp = 111-1 12°C; FTIR (ATR) v _max : 2950, 2863, 1760, 1704, 1615 and 1509 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.14 (s, 3-oxobutyl CH ₃ ), 2.30 (s, 2-OAc), 3.80 (s, 3-OCH ₃ ), 6.76 (s, 1-H), 6.89 (s, 4-H). Anal. Calcd. for C ₂₁ H ₂₆ O ₅ : C, 70.37; H, 7.31. Found: C, 70.44; H, 7.38. Example 59

(1 ObS, 4aS, 4bS)-8-Hydroxy-9-methoxy-3, 4, 5, 6,11,12,1 Ob, 4a, 4b-nonahydrochrysen-2-one (33a)

[000160] Under nitrogen, the diketone compound (32a, 3 g, 8.4 mmol) was cyclized in methanol (100 ml) with 10% KOH solution (40 ml) at reflux for 2.5 hours. Analysis by TLC (5%acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was quenched with water, acidified with 10% HCl, concentrated in vacuo, and extracted with EtOAc (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo to give the cyclized compound (33a, 2.4 g, 99 %) as a yellow solid, mp - 211-213 ⁰ C; FTIR (ATR) v _max : 3267, 2923,

2863, 1658, and 1505 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 3.86 (s, 9-OCH ₃ ), 5.91 (s, 1-H) 6.64 (s, 7-H), 6.78 (s, 10-H). Anal. Calcd. for C ₁₉ H ₂₂ O ₃ : C, 76.48; H, 7.43. Found: C, 76.48; H, 7.48. Example 60

(1 OaS, 1 ObS, 4bS)-3-Methoxy-8-oxo-5, 6, 9,10,11,12,1 Oa, 1 Ob, 4b-nonahydrochrysen-2-yl acetate (34a)

[000161] Under nitrogen, the 3-hydroxy compound (33a, 0.62 g, 2.0 mmol) was acetylated in pyridine (50 ml) with acetic anhydride (10 ml, 105.8 mmol) at room temperature overnight. Analysis by TLC (3% acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was quenched with methanol and concentrated in vacuo. Purification by flash chromatography (3% acetone/CH ₂ Cl ₂ ) and subsequent crystallization from ether gave the 2-acetate derivative (34a, 0.36 g, 51%) as a white solid, mp = 139 ⁰ C; FTIR (ATR) v _max : 2916, 2860, 1759, 1665, 1615 and 1508 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.30 (s, 2-OAc), 3.81 (s, 3-OCH ₃ ), 5.90 (s, 7-H) 6.76 (s, 1-H), 6.89 (s, 4-H). Anal. Calcd. for C ₂₁ H ₂₄ O ₄ ^• 1/5H ₂ O: C, 73.32; H, 7.15. Found: C, 73.37; H, 7.01. Example 61

(4bS, 10bR)-8-Hydroxy-3-methoxy-5, 6,11,12,10b, 4b-hexahydrochrysene-2-yl acetate (35a)

[000162] Under nitrogen, compound (34a, 2.3 g, 5.3 mmol) in CH ₃ CN (150 ml) was aromatized with CuBr ₂ (1.4 g, 6.3 mmol) at room temperature for 24 hours. Analysis by TLC (5% acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was quenched with water and extracted with EtOAc (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (5% acetone/CH ₂ Cl ₂ ) and subsequent crystallization from ether gave the hexahydrochrysene derivative (35a, 1.6 g, 70%) as a white solid, mp = 199-200 ⁰ C; FTIR (ATR) v _max : 3455, 2928, 2839, 1745, 1610, and 1503 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 2.33 (s, 2-OAc), 3.84 (s, 3-OCH ₃ ), 5.13 (s, 8- OH), 6.59 (d, J = 2.7 Hz, 7-H), 6.62 (dd, J ₁ = 8.25 Hz, J ₂ = 3.0 Hz, 9-H), 6.80 (s, 1-H), 6.96 (s, 4-H), 7.20 (d, J = 8.4 Hz, 10-H). Anal. Calcd. for C ₂ ]H ₂₂ O ₄ : C, 74.54; H, 6.55. Found: C, 74.11 ; H, 6.58.

Example 62

(4bS, 10bR)-3-Methoxy-5, 6,11,12,10b, 4b-hexahydrochrysene-2, 8-diol (36a) [000163] Under nitrogen, the 3-acetate compound (35a, 0.17 g, 0.5 mmol) was hydrolyzed in MeOH/H ₂ O (60 ml, 5:1) with potassium carbonate (1.9 g, 13.7 mmol) at room temperature for 2 hours. Analysis by TLC (5% acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was quenched with water and extracted with EtOAc (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (5% acetone/CH ₂ Cl ₂ ) and subsequent crystallization from ether/hexanes gave compound (36a, 130 mg, 88%) as a white solid, mp = 225 ⁰ C; FTIR (ATR) v _max : 3458, 3312, 2922, 2829, 1614, 1584, 1497, and 1463 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 3.89 (s, 3-OCH ₃ ), 4.74 (s, 2-OH), 5.49 (8-OH), 6.63 (d, J = 3.0 Hz, 7-H), 6.68 (dd, J ₁ = 8.4 Hz, J ₂ = 3.0 Hz, 9-H), 6.71 (s, 1-H), 6.87 (s, 4-H), 7.25 (d, J = 8.4 Hz, 10- H). Anal. Calcd. for Ci ₉ H ₂₀ O ₃ - 1/2H ₂ O: C, 74.73; H, 6.93. Found: C, 74.82; H, 6.70. Example 63

(4bS, 10bR)-2, 8-bis(Methoxymethoxy)-3-methoxy-5 , 6,11,12,1 Ob, 4b-hexahydrochrysene (37a)

[000164] Under nitrogen, the 2,8-diol derivative (36a, 1 g, 3.4 mmol) in THF (100 ml) was treated with N,N-diisopropylethylamine (2.1 ml, 5.75 mmol) and chloromethyl methyl ether (6 ml, 79 mmol) and the reaction mixture stirred at 65-7O ⁰ C overnight. Analysis by TLC (5% acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was cooled to room temperature, quenched with 20% NH ₄ Cl solution, concentrated in vacuo, and extracted with EtOAc (3x). The organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (5% acetone/CH ₂ Cl ₂ ) gave the dimethoxymethyl ether derivative (37a, 1.1 g, 88 %) as a low melting solid, mp = 83 - 84 ⁰ C; FTIR (ATR) V _108x : 2908, 2834, 1606, and 1498 cm ^"1 . NMR (300 MHz, CDCl ₃ ), 5 _H (ppm): 3.48 (s, 8-OCH ₂ OCH ₃ ), 3.52 (s, 2-OCH ₂ OCH ₃ ) 3.88 (s, 3-OCH ₃ ), 5.16 (s, 8- OCH ₂ OCH ₃ ), 5.21 (s, 2-OCH ₂ OCH ₃ ), 6.84 (d, J= 2.4 Hz, 7-H), 6.88 (dd, J ₁ = 8.4 Hz, J ₂ = 2.7 Hz, 9-H), 6.92 (s, 1-H), 6.93 (s, 4-H). 7.30 (d, J = 8.4 Hz, 10-H). Anal. Calcd. for C ₂₃ H ₂₈ O ₅ : C, 71.85; H, 7.34. Found: C, 71.68; H, 7.32.

Example 64

(1 ObS , 4bR)-2, 8-bis(Methoxymethoxy)-9-methoxy-5, 6,11,12,10b, 4b-hexahydrochrysen-3- ol (38a)

[000165] Under nitrogen, the dimethoxymethyl ether derivative (37a, 0.5 g, 1.3 mmol) in THF (10 ml) at -78 ⁰ C was treated with the dropwise addition of sec-BuLi (1.4 M/cyclohexane, 1.8 ml, 2.52 mmol) and the reaction stirred for 3 hours. Trimethyl borate (0.6 ml, 5.4 mmol) was added dropwise and the reaction mixture stirred for 20 minutes. The reaction mixture was warmed to O ⁰ C and quenched with 20 % NH ₄ Cl solution (10 ml) and stirred at room temperature for 1 hour. Sodium perborate tetrahydrate (0.8 g, 5.2 mmol) was added and the reaction mixture stirred at room temperature overnight. The mixture was extracted with EtOAc (3x), the organic fractions were washed with water (3x) and brine, combined, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. Purification by flash chromatography (5% acetone/CH ₂ Cl ₂ ) gave a mixture of the starting material (0.14 g) and the 16-hydroxy derivative (38a, 0.31 g, 84 %), which crystallized from ether/hexanes as a white solid, mp = 117-119 ⁰ C; FTIR (ATR) v _max : 3440, 2909, 2838, 1608, and 1502 cm- ¹ . NMR (300 MHz, CDCl ₃ ), 6 _H (ppm): 3.50 (s, 2-OCH ₂ OCH ₃ ), 3.52 (s, 8-OCH ₂ OCH ₃ ) 3.87 (s, 9-OCH ₃ ), 5.15 (s, 2-OCH ₂ OCH ₃ ), 5.21 (s, 8- OCH ₂ OCH ₃ ), 5.99 (3-OH), 6.86 (s, 1-H), 6.91 (d, J = 8.4 Hz, 4-H), 6.92 (s, 7-H), 6.98 (s, 10-H). Anal. Calcd. for C ₂₃ H ₂₈ O ₆ : C, 69.98; H, 7.05. Found: C, 69.83; H, 6.99. Example 65

(4bS, 10bR)-2, 8-bis(Methoxymethoxy)-3, 9-dimethoxy-5, 6,11,12,1 Ob, 4b- hexahydrochrysene (39a)

[000166] Under nitrogen, the 16-hydroxy derivative (38a, 0.5 g, 1.24 mmol) in DMF (20 ml) was methylated with methyl iodide (5 ml, 80.3 mmol) in the presence of potassium carbonate (2 g, 14.5 mmol) and tetrabutyl ammonium iodide (20 mg) at room temperature overnight. Analysis by TLC (3% acetone/CH ₂ Cl ₂ ) indicated an incomplete reaction Additional methyl iodide (5 ml) was added and the reaction mixture stirred another 2 hours. Analysis by TLC indicated no further progress. The reaction mixture was quenched with brine and extracted with EtOAc (3x). The organic fractions were washed with brine, combined, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (3% acetone/C^Ch) gave the dimethoxy compound (39a, 0.38 g, 73 %) as a yellow solid, mp = 151-153 ⁰ C; FTIR (ATR) v _max : 2944, 2913, 2827, 1607, and 1510 cm ^"1 . NMR (300 MHz, CDCl ₃ ), δ _H (ppm): 3.53 (s, 2,8-

OCH ₂ OCH ₃ ), 3.89 (s, 3,9-OCH ₃ ), 5.22 (s, 2,8-OCH ₂ OCH ₃ ), 6.94 (s, 1,4,7,10-H). Anal. Calcd. for C ₂₄ H ₃₀ O ₆ : C, 69.55; H, 7.30. Found: C, 69.56; H, 7.23. Example 66

(4bS, 10bR)-3, 9-Dimethoxy-5, 6,11,12,1 Ob, 4b-hexahydrochrysene-2, 8-diol (40a) [000167] Under nitrogen, the dimethoxymethyl ether derivative (39a, 0.5 g, 1.2 mmol) in THF (30 ml) was hydrolyzed with 6M HCl (10 ml) at room temperature overnight. Analysis by TLC (3% acetone/CH ₂ Cl ₂ ) indicated a complete reaction. The reaction mixture was quenched with water and extracted with EtOAc (3x). The organic fractions were washed with water (3x) and brine, dried over Na ₂ SO ₄ , filtered, and concentrated in vacuo. Purification by flash chromatography (5% acetone/CH ₂ Cl ₂ ) and subsequent crystallization from MeOH/EtOAc gave compound (40a, 0.16 g, 40%) as a white solid. mp = 295 ⁰ C (dec); FTIR (ATR) v _max : 3511, 3346, 2958, 2909, 2841, 1622, and 1500 cm ^" '. NMR (300 MHz, (CD ₃ ) ₂ SO), δ _H (ppm): 3.73 (s, 3,9-OCH ₃ ), 6.50 (s, 1,7-H), 6.88 (s, 4,10-H). Anal. Calcd. for C ₂₀ H ₂₂ O ₄ - V. H ₂ O: C, 72.60; H, 6.85. Found: C, 72.65; H, 6.67. [000168]

Table 1. MM3 Relative energies of trans- and gsuche g ⁺ conformations of 18a-homo steroids * Dihedral angle for the CH-ClS-Cl δ-Cl δa bond.

Table 2. Selected proton chemical shifts for 10a, 11a, and 12a.

Table 3. 18-Methyl and 18a-Methyl chemical shifts • Data taken from reference 3 and 6

[000169] All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the methods described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.