KR20150135547A - Estrogen receptor ligands and methods of use thereof - Google Patents

Abstract

The present invention relates to a method of reducing testosterone levels by decreasing luteinizing hormone (LH) in a male subject, or independently of LH levels, a method of treating, inhibiting, reducing the incidence, reducing the severity, or inhibiting advanced prostate cancer, It is about palliative treatment of cancer.

Description

ESTROGEN RECEPTOR LIGANDS AND METHODS OF USE THEREOF < RTI ID = 0.0 >

The present invention relates to a method of reducing testosterone levels by decreasing luteinizing hormone (LH) in a male subject, or independently of LH levels, a method of treating, inhibiting, reducing the incidence, reducing the severity, or inhibiting advanced prostate cancer, It is about palliative treatment of cancer.

Estrogen is a group of resistance and synthetic hormones that are important for tissue and bone maintenance and are used for this purpose. Estrogen is an endocrine modulator in the cellular processes involved in the development and maintenance of the reproductive system. The role of estrogen in reproductive biology is well known for the prevention of postmenopausal flushing and postmenopausal osteoporosis prevention. Estradiol is a major endogenous human estrogen, found in both women and men.

The biological action of estrogen and antiestrogen appears through two intracellular receptors, the estrogen receptor alpha (ERa) and the estrogen receptor beta (ERβ). Endogenous estrogens are typically potential activators of both receptor subtypes. For example, estradiol acts as an ERa agonist in many tissues, including breast, bone, cardiovascular, and central nervous system tissues. Selective estrogen receptor modulators typically work differently in different tissues. For example, SERM may be an ERa antagonist in the breast, but may be a partial ERa agonist in the uterus, bone and cardiovascular system. Compounds acting as estrogen receptor ligands are therefore useful in the treatment of a variety of conditions and disorders.

Prostate cancer is one of the most frequently diagnosed non-skin cancers among men in the United States, the second most common cause of cancer deaths with more than 180,000 new cases, and is expected to close to 29,000 deaths this year. Patients with advanced prostate cancer are typically treated with luteinizing hormone-releasing hormone (LHRH) agonists or with bilateral testicular resection (ADT). Androgen blockers not only reduce testosterone, but also reduce estrogen levels because estrogen is derived from the aromatization of testosterone and the level of testosterone is deficient by ADT. Estrogen deficiency induced by androgen blockers results in significant side effects, including facial flushing, gynecomastia and mastectomy, bone loss, bone loss and strength reduction, osteoporosis and life-threatening fractures, deleterious lipid changes and cardiovascular disease, Infarction, and depression and other mood changes. It is believed that many estrogen deficiency side effects of ADT are mediated by ERα.

Looprolide acetate (Lupron®) is a nonapeptide analog of naturally occurring gonadotropin-releasing hormone (GnRH or LH-RH). Loopride Acetate is an LH-RH superagonist that eventually inhibits LH secretion by the pituitary gland. Looprolide acetate acts as a potential inhibitor of gonadotropin secretion, inhibiting ovarian and testicular steroid production. In humans, the administration of looped lead acetate leads to an initial increase in the circulating levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH), leading to gonadal hormones (testosterone and dihydrootestosterone in males, and estrone And estradiol) levels. However, continuous administration of the looped lead acetate results in a decrease in LH and FSH levels. In men, testosterone is reduced to castrate levels (<50 ng / dL). In premenopausal women, estrogen is reduced to postmenopausal levels. Testosterone is known to stimulate prostate cancer cells. Therefore, inhibition of testosterone secretion or inhibition of testosterone function is necessary for the treatment of prostate cancer. Looprolide acetate can be used for LH inhibition, which treats prostate cancer by reducing and lowering serum testosterone to a steady level.

Prior to the introduction of the LHRH agonist, the level of steroid testosterone was achieved by increasing estrogenic activity in the pituitary gland through estrogen, primarily diethylstilbestrol (DES). DES is equally effective as an LHRH agonist in suppressing testosterone to a steady level. DES did not show flushing or bone loss, but showed a higher gynecomastia ratio than ADT treated with LHRH agonist. Unfortunately, very potent pure estrogens such as DES and estradiol are often associated with high risk of serious cardiovascular and thromboembolic complications and have limited their clinical use.

The compounds of the present invention prevent the increased risk of thrombosis and inhibit testosterone levels to a steady level without causing bone loss, facial flushing and / or gynecomastia, which can be used for the treatment of prostate cancer.

Summary of the Invention

In one embodiment, the invention provides a method of reducing total serum testosterone levels in a male subject, comprising administering a therapeutically effective amount of a compound of formula I-XII as set out below.

In one embodiment, the present invention provides a method of reducing total serum testosterone levels in a male subject comprising administering a therapeutically effective amount of a compound of formula I-XII as set forth below, wherein said total serum testosterone reduction is selected from the group consisting of serum progesterone Lt; RTI ID = 0.0 &gt; hormone &lt; / RTI &gt; level.

In one embodiment, the present invention provides a method of reducing total serum testosterone levels in a male subject comprising administering a therapeutically effective amount of a compound of formula I-XII as set forth below, wherein said total serum testosterone reduction is selected from the group consisting of serum progesterone Lt; RTI ID = 0.0 &gt; hormone &lt; / RTI &gt; levels.

In one embodiment, the present invention is directed to a method of reducing total serum testosterone levels in a male subject comprising administering a therapeutically effective amount of a compound of formula I-XII as set out below, wherein said compound administration is selected from the group consisting of androgen- ), Wherein the subject has prostate cancer. The present invention also provides a method for preventing or treating the occurrence of side effects associated with prostate cancer.

In one embodiment, the present invention provides a method for androgen blockade therapy in a subject comprising administering a therapeutically effective amount of a compound of formula I-XII as set forth below. In another embodiment, .

In one embodiment, the invention provides a method of treating, inhibiting, reducing the incidence, severity, or inhibiting progressive prostate cancer, comprising administering a therapeutically effective amount of a compound of formula I-XII as set forth below .

In one embodiment, the invention provides a method of palliative treatment of advanced prostate cancer, comprising administering a therapeutically effective amount of a compound of formula I-XII as set forth below.

The compounds of the present invention prevent the increased risk of thrombosis and inhibit testosterone levels to a steady level without causing bone loss, facial flushing and / or gynecomastia, which can be used for the treatment of prostate cancer.

The gist of the present invention is specifically described and explicitly claimed in the conclusion of the specification. The invention, however, both as to organization and method of operation, may best be understood by reference to the following detailed description, as well as the objects, features and advantages thereof, as well as the accompanying drawings.
Figure 1 shows the levels of serum testosterone (solid line) and total androgen (dashed line) after oral administration of compound IV at 30 mg / kg daily (first administered at day 0) in intact male monkeys. (See Example 8)
Figure 2 shows the levels of testosterone in intact rat treated with compound IV (0.3, 1, 10, 30 mg / kg). ^I & lt ^{; /} RTI ^&gt; Uninfected Beihikle control group. BLOQ values are plotted at the limit of quantitation 0.08 ng / mL. (See Example 9).
Figure 3 shows the inhibitory effect of compound IV on 17 [beta] -HSD5 enzyme activity. (See Example 12).
Figure 4 shows in vitro aggregation of human platelets in the presence of DES, 17 [beta] -estradiol (E2), and Compound IV. Platelet-rich plasma (PRP) was incubated with Beckle, E2, DES, or Compound IV for 30 seconds before inducing aggregation to 0.3 units of thrombin. Aggregation was monitored for 5 minutes and expressed as a percentage of the Beihikle control (see Example 13).
Figure 5 is a comprehensive synthesis scheme for the preparation of compounds II-XII (see Example 1).
Figure 6 is a synthesis schematic for the preparation of compound IV (see Example 2).
Figure 7 is a synthesis schematic for the preparation of compound VI (see Example 3).
Figure 8 is a synthesis schematic for the preparation of compounds IX and X (see Example 5).
Figure 9 shows testosterone levels at 24 hours, 72 hours and 168 hours at doses of 3 mg / kg, 10 mg / kg and 300 mg / kg in intact rat treated with compound IV. (See Example 9).
10 shows the LH levels of intact testis-extracted (ORX) rats treated with Compound IV at doses of 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 10 mg / kg and 30 mg / ), FSH levels (Figure 10b), testosterone levels (Figure 10c), prostate weight levels (Figure 10d), seminal vesicle weight levels (Figure 10e), and anorectal mass (Figure 10f). ^I & lt ^{; /} RTI ^&gt; Uninfected Beihikle control group. ^{O &lt}; / RTI ^&gt; ORX &lt; / RTI &gt; BLOQ values are plotted at the limit of quantitation of 0.08 ng / mL (see Example 9).
Figure 11 shows the intact prostate size in ORX rats by administering Compound IV (Figure 11A) and DES (Figure 11B) at different doses (see Example 15).
Figure 12 shows the difference between DES and Compound IV; DES cross-reacts with the glucocorticoid receptor (GR), whereas compound IV does not (Figure 12a); DES cross-reacts with the androgen receptor (AR). This mildly stimulates and mildly inhibits AR action (i.e. it is a partial agonist / antagonist), whereas compound IV is not (Fig. 12b); DES eliminates the estrogen-related receptor (ERR) transfer activity, whereas compound IV does not (Figure 12c). (See Example 15).
Figure 13 shows the effect of Compound IV on facial flushing weakness at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, and 30 mg / kg of the morphine withdrawal model. N = 7 animals per group. 17 [beta] -E2 was used at 5 mg / kg in 100% DMSO (see Example 14).
Figure 14 shows the dose-dependent decrease in monkey body weight (kg) from 100 mg / kg to ~ 20% by administering compound IV for 91 days. No sign of gynecomastia or hyperestrogenicity was observed. (See Example 16)
Figure 15 shows the dose-dependent decrease in serum testosterone levels (ng / mL) after daily oral administration of compound IV compared to the positive control (LHRH agonist). The dotted line represents testosterone levels in chemically castrated patients and the thick dashed lines represent the testosterone levels of surgically sterile monkeys (see Example 16).
Figure 16 shows dose-dependent prostate-specific antigen (PSA) levels in monkeys by administering compound IV at baseline and at 28 days. PSA levels were significantly reduced with Compound IV treatment (see Example 16).
Figure 17 shows the dose-dependent prostate volume using monkey intussusum ultrasonography (TRUS) by administering compound IV at 6 weeks compared to the positive control (LHRH agonist). (See Example 16)
Figure 18 shows the dose-dependent organ weights (prostate, seminal vesicles, and testes) as a percentage of control monkeys on day 90 by administering compound IV (Fig. 18A). Prostate weight at 13 weeks post-mortem in monkey after oral administration of Compound IV daily (Figure 18b). (See Example 16).
Figure 19 shows the dose-dependent average total testosterone level (nmol / L) in humans for a period of 1-11 days by administering Compound IV (100 mg, 300 mg, 600 mg and 1000 mg). (See Example 17)
Figure 20 shows the dose-dependent average LH level (IU / L) in humans for a period of 1-10 days by administering Compound IV (100 mg, 300 mg, 600 mg and 1000 mg). (See Example 17)
Figure 21 shows the dose-dependent average free testosterone level (pg / mL) in humans for a period of 1-10 days by administering Compound IV (100 mg, 300 mg, 600 mg and 1000 mg) Reference)
Figure 22 shows the dose-dependent average PSA level (占 퐂 / L) in humans for a period of 1-10 days by administering Compound IV (100 mg, 300 mg, 600 mg and 1000 mg). (See Example 17)
Figure 23 shows dose-dependent serum testosterone levels (ng / mL) in intact rat after 14 days recovery of compound IV administration. ^I represents a P &lt; 0.05 vs uninfected control group (see Example 10)
It will be appreciated that for simplicity and clarity of illustration, components shown in the figures are not necessarily drawn to scale. For example, the scale of some components may be exaggerated relative to other components for clarity. Further, where considered appropriate, reference numerals will be repeated throughout the drawings to indicate corresponding or analogous features.

In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. However, it will be understood by those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures and components are not described so as not to obscure the present invention.

In one embodiment, the compounds described herein and / or compositions comprising them will be useful for reducing total serum testosterone levels in male subjects.

In one embodiment, the compounds described herein and / or compositions comprising them will be useful for reducing total serum testosterone levels in male subjects, wherein the total serum testosterone reduction occurs by a decrease in serum luteinizing hormone (LH) levels .

In one embodiment, in one embodiment, the compounds described herein and / or compositions comprising the same will be useful in reducing total serum testosterone levels in a male subject, wherein the total serum testosterone reduction is a decrease in serum luteinizing hormone levels It is irrelevant.

In one embodiment, the present invention includes a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound represented by the structure of Formula I, or an isomer thereof, a pharmaceutically acceptable salt, a pharmaceutical product, a polymorph, a hydrate, , A method of reducing total serum testosterone levels in a male subject is provided:

In this formula,

Y is C (O) or CH ₂ ;

R ₁ and R ₂ are independently selected from the group consisting of hydrogen, halogen, hydroxyl, alkoxy, cyano, nitro, CF ₃ , N (R) ₂ , sulfonamide, SO ₂ R, alkyl, haloalkyl, aryl, O-Alk-NR ₅ R ₆ Or O-Alk-heterocycle, wherein said heterocycle is optionally aromatic and is a 3-7 membered substituted or unsubstituted heterocycle ring;

R ₃ and R ₄ are independently selected from the group consisting of hydrogen, halogen, hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF ₃ , NHCOR, N (R) ₂ , sulfonamide, SO ₂ R, Or a protected hydroxyl group;

R is alkyl, trihalomethyl alkyl, hydrogen, haloalkyl, dihalo, CH ₂ F, CHF ₂ , CF ₃ , CF ₂ CF ₃ , aryl, phenyl, halogen, alkenyl, CN, NO ₂ , or OH;

R ₅ and R ₆ are independently hydrogen, phenyl, alkyl of 1-6 carbon atoms, 3-7 membered cycloalkyl, 3-7 membered heterocycle, 5-7 membered aryl; R &lt; ₅ &gt; and R &lt; ₆ &gt; together with the nitrogen atom form a 3-7 membered ring;

j and k are independently 1 to 4;

Alk is linear alkyl of 1-7 carbons, branched alkyl of 1-7 carbons, or cyclic alkyl of 3-8 carbons.

In an additional embodiment of the method described herein, the compound of formula I is represented by formula IA:

Wherein R ₁ , R ₂ , R ₃ , R ₄ , j and k are as defined for formula I.

In one embodiment, the present invention is directed to a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound represented by a compound of formula II, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention is directed to a pharmaceutical composition comprising a therapeutically effective amount of a compound represented by the formula III compound, or an isomer thereof, a pharmaceutically acceptable salt, a pharmaceutical product, a polymorph, a hydrate or a combination thereof , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention includes a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound represented by the formula IV, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or combination thereof , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention is directed to a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound represented by the formula V compound, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or combination thereof , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention includes a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound represented by a compound of formula VI, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention includes a method of treating or preventing a disease or disorder comprising administering a therapeutically effective amount of a compound represented by the formula VII compound, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or a combination thereof , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention is directed to a method of treating or preventing a disease or disorder comprising administering a therapeutically effective amount of a compound represented by the formula VIII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or combination thereof , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention is directed to a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound represented by the formula IX, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or combination thereof , A method of reducing total serum testosterone levels by reducing the level of luteinizing hormone (LH) in a male subject with prostate cancer is provided:

In one embodiment, the present invention includes a method of treating a patient suffering from a disease or disorder comprising administering a therapeutically effective amount of a compound represented by a compound of formula X, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention is directed to a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound represented by the formula XI, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention includes a method of treating a patient suffering from a disease or disorder comprising administering a therapeutically effective amount of a compound represented by the formula XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or combination thereof , A method of reducing total serum testosterone levels in a male subject is provided:

In one embodiment, the present invention provides a method of treating or preventing a disorder or condition selected from the group consisting of: administering a therapeutically effective amount of a compound of formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, A method of reducing total serum testosterone levels in a male subject is provided. In another embodiment, the male subject has prostate cancer. In another embodiment, the total serum testosterone is reduced to less than about 100 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 50 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 25 ng / dL.

In one embodiment, the present invention is directed to a method of treating a male patient comprising administering a therapeutically effective amount of a compound of formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, Wherein the total serum testosterone reduction is caused by a decrease in serum luteinizing hormone (LH) levels. In another embodiment, the male subject has prostate cancer. In another embodiment, the total serum testosterone is reduced to less than about 100 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 50 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 25 ng / dL.

In one embodiment, the present invention is directed to a method of treating a male patient comprising administering a therapeutically effective amount of a compound of formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, Wherein the free serum testosterone reduction is caused by a decrease in serum luteinizing hormone (LH) levels, as a method for reducing free serum testosterone levels in a subject. In another embodiment, the male subject has prostate cancer.

In one embodiment, the present invention is directed to a method of treating a male patient comprising administering a therapeutically effective amount of a compound of formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, Wherein the total serum testosterone reduction is independent of a decrease in serum luteinizing hormone (LH) levels. In another embodiment, the male subject has prostate cancer. In another embodiment, the total serum testosterone is reduced to less than about 100 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 50 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 25 ng / dL.

In one embodiment, the present invention is directed to a method of treating a male patient comprising administering a therapeutically effective amount of a compound of formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, A method of reducing free serum testosterone levels in a subject, wherein said free serum testosterone reduction is independent of a decrease in serum luteinizing hormone (LH) levels. In another embodiment, the male subject has prostate cancer.

In one embodiment, the invention provides a method of reducing total serum testosterone or free serum testosterone levels in a male subject, wherein the male subject has prostate cancer. In another embodiment, the subject has advanced prostate cancer.

In one embodiment, the decrease in serum concentration of testosterone is reversible and returns to baseline levels after treatment with the compounds of the invention.

In another embodiment, the serum concentration of testosterone is reversible after treatment with compound IV according to FIG. 23 and Example 10.

In one embodiment, the present invention provides a method of treating or preventing a disorder or condition selected from the group consisting of: administering a therapeutically effective amount of a compound of formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, A method of reducing total serum testosterone levels in a male subject is provided. In another embodiment, the total serum testosterone is reduced to less than about 100 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 50 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 25 ng / dL. In another embodiment, the total serum testosterone is reduced to less than about 75 ng / dL. In another embodiment, the total serum testosterone is reduced to between about 75 ng / dL -100 ng / dL. In another embodiment, the total serum testosterone is reduced to between about 50 ng / dL-75 ng / dL. In another embodiment, the total serum testosterone is reduced to between about 40 ng / dL and 50 ng / dL. In another embodiment, the total serum testosterone is reduced to between about 25 ng / dL and 50 ng / dL. In another embodiment, the total serum testosterone is reduced to between about 40 ng / dL and 60 ng / dL.

Testosterone can be measured as serum levels of "free" (ie, biologically available and unbound) or "total" (including percentages of conjugated unavailable proteins). Men over 40 years of age who do not have prostate cancer show a lower testosterone level of less than 250 ng / dL (<8.7 nmol / L) or a lower free testosterone level of less than 0.75 ng / dL (<0.03 nmol / L).

In one embodiment, the method of the present invention provides a method of reducing total serum and / or free testosterone levels by reducing LH levels in a male subject with prostate cancer, regardless of a decrease in LH levels. In other embodiments, the testosterone level will be a decrease from the pretreatment level. In another embodiment, the total serum testosterone level is reduced to less than 100 ng / dL. In another embodiment, the total serum testosterone level is reduced to less than 50 ng / dL. In another embodiment, the total serum testosterone level is reduced to less than 25 ng / dL. In another embodiment, the free serum testosterone level is reduced to 2 ng / dL or less. In another embodiment, the free serum testosterone level is reduced to less than 1 ng / dL. In another embodiment, the free serum testosterone level is reduced to less than 0.5 ng / dL. In another embodiment, the free serum testosterone level is reduced to less than 0.25 ng / dL.

Methods for measuring free serum testosterone levels and total serum testosterone levels include monitoring testosterone levels during treatment by blood testing. Total testosterone is a combination of circulating testosterone and free / non-binding hormone bound to carrier proteins (albumin, SHBG, transcotin, transferrin). Total testosterone levels may be influenced by several factors including the level of protein in the blood that transports the hormone into the body, age, obesity, and interference associated with commonly used test methods.

Useful methods for measuring free testosterone (FT) can be either complex (equilibrium dialysis and free testosterone calculations (CFT)) or simple (commercial FT kit "Coat-A-Count") using an analogous tracer. In other embodiments, measurements of total testosterone and free testosterone serum levels can be achieved by measuring total testosterone and SHBG (e.g., Irma-Count, DPC) simultaneously and then measuring free testosterone values (CFT). In other embodiments, measurement of total testosterone and free testosterone may be in accordance with the knowledge in the art.

In one embodiment, the present invention provides a method of treating a compound of Formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, A method of reducing total serum testosterone levels or free serum testosterone levels in a male subject, comprising administering an effective amount of a compound of formula In other embodiments, total or free serum testosterone reduction occurs by a decrease in serum luteinizing hormone (LH) levels. In another embodiment, the decrease in total or free serum testosterone levels is independent of the decrease in serum luteinizing hormone levels.

The methods of the invention include administering a combination of compounds of the invention and other forms of ADT. In one embodiment, other forms of ADT include an LHRH agonist. In another embodiment, the LHRH agonist is selected from the group consisting of looprolide acetate (Lupron®) (US Pat. Nos. 5,480,656, 5,575,987, 5,631,020, 5,643,607, 5,716,640, 5,814,342, 6,036,976, ) Or goserelin acetate (Zoladex®) (US Pat. Nos. 7,118,552; 7,220,247; 7,500,964, all incorporated herein by reference). In one embodiment, other forms of ADT include an LHRH antagonist. In another embodiment, the LHRH antagonist comprises degalelix. In one embodiment, other forms of ADT include anti-androgens. In another embodiment, the anti-androgen includes bicalutamide, flutamide, panestelide, dutasteride, MDV 3100M, neilutamide, chlormadinone, or a combination thereof.

In one embodiment, the methods of the invention comprise administering a therapeutically effective amount of an anti-androgen and a compound of the invention. In one embodiment, the methods of the invention comprise administering a therapeutically effective amount of an LHRH agonist and a compound of the invention. In one embodiment, the methods of the invention comprise administering a therapeutically effective amount of an anti-androgen, an LHRH agonist, and a compound of the invention.

In one embodiment, the present invention provides a method for the treatment and prophylaxis of lupus erythematosus (LH) in a male subject having prostate cancer for the purpose of producing androgen-blocking therapy (ADT), comprising administering a therapeutically effective amount of a compound of formula IA, ) Level or a decrease in total serum testosterone levels and / or free testosterone levels independent of a decrease in luteinizing hormone levels. In another embodiment, the compound is compound IV.

In another embodiment, the present invention provides a method of treating a disorder or condition selected from the group consisting of: administering a therapeutically effective amount of a compound of formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, Provides a method for androgen-blocking therapy (ADT) in a subject. In another embodiment, the subject has prostate cancer. In another embodiment, the compound is compound IV.

In another embodiment, ADT is used for prostate cancer treatment, delayed progression of prostate cancer, or prevention and / or treatment of prostate cancer recurrence.

In one embodiment, the invention provides a method of preventing, treating and / or preventing recurrence of prostate cancer, comprising administering a compound of the invention. In other embodiments, the compounds of the present invention may be administered in combination with an LHRH analog, a reversible anti-androgen (such as bicalutamide or flutamide), an anti-estrogen, an anticancer, a 5-alpha reductase inhibitor, an aromatase inhibitor, progestin, Androgen receptor modulators (SARMS), or other agents acting through nuclear hormone receptors.

In one embodiment, the invention provides a method of treating prostate cancer, comprising administering a compound of Formula IA, I-XII, which is independent of LH level reduction or a decrease in LH level, and treatment of total serum testosterone and / It provides a method of reducing testosterone levels. In another embodiment, compound IV is administered.

Androgen blockers not only reduce testosterone, but also estrogen levels are reduced because estrogen is derived from the direction of testosterone. Estrogen deficiency induced by androgen blockers is associated with a number of complications including facial flushing, gynecomastia and mastectomy, bone loss, bone and strength reduction, osteoporosis, osteopenia, and life-threatening fractures, deleterious lipid changes and higher cardiovascular disease and myocardial infarction, Causing significant side effects including loss of libido, impotence, muscle mass loss (myopathy), fatigue, cognitive dysfunction, and depression and other mood changes.

In other embodiments, the invention provides methods of treating diseases, disorders or conditions associated with ADT. In other embodiments, the invention provides a method of treating a disease, disorder or condition associated with a testosterone deficiency. Each disease, disorder or condition represents a separate embodiment of the present invention.

In one embodiment, the present invention is directed to a method of treating or preventing a disease or condition comprising administering a therapeutically effective amount of a compound of Formula IA, I-XII, or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, Or a pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or a combination thereof, of a compound of formula IA, I-XII, or an isomer thereof, a pharmaceutically acceptable salt, Inhibiting, causing, reducing, inhibiting or treating the occurrence of side effects associated with ADT, the subject having prostate cancer. In other embodiments, the total or free serum testosterone level reduction is independent of the decrease in LH level or the decrease in LH level.

In one embodiment, administration of a compound of the present invention inhibits, reduces, inhibits, or treats the occurrence of typical side effects associated with traditional androgen-blocking therapy (ADT). In another embodiment, the subject has prostate cancer. Prevention and / or reduction of such side effects are for placebo or control. In one embodiment, typical side effects associated with the traditional androgen blockade regimen (ADT) include facial flushing, gynecomastia, decreased bone density, and increased fracture. In other embodiments, the administration of the compounds of the present invention prevents the occurrence of facial flushes found in the use of traditional forms of androgen-blocking therapy (ADT). In other embodiments, the administration of the compounds of the present invention prevents the occurrence of gynecomastia found in conventional androgen-blocking therapy (ADT). In other embodiments, administration of the compounds of the present invention prevents the occurrence of a decrease in bone density (BMD) found using conventional androgen-blocking therapy (ADT). In another embodiment, the administration of a compound of the present invention prevents the occurrence of an increased fracture found when using traditional androgen-blocking therapy (ADT). In other embodiments, the increase in fracture is a pathological fracture, a non-traumatic fracture, a vertebral fracture, a non-vertebral fracture, a new morphological fracture, a clinical fracture, or a combination thereof.

In one embodiment, the term "traditional androgen-blocking therapy" refers to testicular resection (surgical castration) in which a surgeon removes testicles. In another embodiment, the term "traditional androgen-blocking therapy" relates to administering a luteinizing hormone-releasing hormone (LHRH) analogue: such a drug reduces the amount of testosterone produced by the testis. Examples of useful LHRH analogs in the United States include looprolides (Lupron, Viadur, Eligard), goserelin (Zoladex), tryptorelin (Trelstar), and histroline (Vantas). In another embodiment, the term "traditional androgen-blocking therapy" relates to administering anti-androgens: anti-androgens block the body's ability to use androgens. Small amounts of androgens are still produced by the adrenal glands after testicular resection or during LHRH analogue treatment. Examples of anti-androgen drugs include flutamide (Eulexin), bicalutamide (Casodex), and neilutamide (Nilandron). In another embodiment, the term "traditional androgen-blocking therapy" refers to administering a luteinizing hormone-releasing hormone (LHRH) antagonist such as Plenaxis; Firmagon is a new LHRH antagonist approved by the FDA in 2008 for the treatment of advanced prostate cancer. In another embodiment, the term "traditional androgen-blocking therapy" refers to administering a 5a-reductase inhibitor such as Proscar and Avodart: a 5a-reductase inhibitor inhibits testosterone Dihydrotestosterone (DHT), an androgen. &Lt; / RTI &gt; In another embodiment, the term "traditional androgen-blocking therapy" refers to administering a testosterone biosynthesis inhibitor such as ketoconazole (Nizoral). In other embodiments, the term "traditional androgen-blocking therapy" refers to administering estrogen such as diethylstilbestrol or 17 [beta] -estradiol.

In one embodiment, the term "facial flushing" refers to a sudden febrile sensation on the upper or the whole of the body, redness of the face and neck, red spots appearing in the chest, back and arms, chills and the like.

In one embodiment, the term "gynecomastia" refers to a benign enlargement of the male breast by the proliferation of mammary gland components, which may or may not be associated with pain. The gynecomastia is clinically defined as the presence of rubbery or rigid mass extending concentrically from the nipple. Symptoms known as caustic gynecomastia or lipopneumonia are characterized by fat accumulation without proliferation. The gynecomastia is usually bilateral or one-sided.

In one embodiment, the method of the invention relates to treating testosterone reduction in a male with prostate cancer or advanced prostate cancer without causing bone loss or facial flushing. In another embodiment, the methods of the invention utilize Compounds IA, I-XII, which do not cause certain side effects, such as bone loss and facial flushing, which are currently common in androgen blockers (ADT) for prostate cancer , But has the potential to reduce testosterone, the primary stimulus for prostate cancer.

In another embodiment, Table 8 (Example 11) below demonstrates that administration of compound IV reduces testosterone without causing bone loss.

In one embodiment, the method of the invention relates to the reduction of testosterone levels, which further treats advanced prostate cancer by administering a compound of formula IA, I-XII. In one embodiment, the methods of the invention relate to a reduction in testosterone levels that further inhibits, reduces, severely reduces or inhibits advanced prostate cancer by administering a compound of Formula IA, I-XII. In one embodiment, the methods of the invention relate to a reduction in testosterone levels, further providing a palliative treatment of advanced prostate cancer by administering a compound of formula IA, I-XII.

In one embodiment, the method of the invention is directed to the treatment of advanced prostate cancer. In one embodiment, the methods of the invention relate to inhibition, reduction of occurrence, severity reduction or inhibition of advanced prostate cancer. In one embodiment, the methods of the invention relate to palliative treatment of advanced prostate cancer. In another embodiment, the methods of the invention utilize compounds of formula IA, I-XII. In another embodiment, the invention is directed to the treatment of advanced prostate cancer. In another embodiment, the invention relates to the inhibition of advanced prostate cancer. In another embodiment, the present invention relates to the reduction of the incidence of advanced prostate cancer. In another embodiment, the present invention relates to a reduction in the severity of advanced prostate cancer. In another embodiment, the invention relates to the inhibition of advanced prostate cancer.

The term "advanced prostate cancer" refers to a metastatic cancer that is derived from the prostate gland and transits widely to the seminal vesicle, pelvic lymph node or bone, or other body parts beyond the prostate gland like surrounding tissue. The pathology of prostate cancer is graded by the Gleason grade from 1 to 5 in order of increasing malignancy. In other embodiments, patients with significant risk of death from progressive disease and / or prostate cancer will be included in the definition, and patients with cancer outside the prostate adventitia with low disease stages, such as IIB, I have.

In one embodiment, the methods provided herein and / or the use of compounds provided herein are effective in providing feedback on the hypothalamus-pituitary-testis axis (HPT axis). Feedback means the ability of a substance produced by one organ or tissue to modulate the activity of another organ or tissue that affects its activity. In one embodiment, feedback on the hypothalamus-pituitary-testis axis (HPT axis) decreases the level of LH. In one embodiment, feedback on the hypothalamus-pituitary-testis axis (HPT axis) reduces total serum testosterone levels. In one embodiment, feedback on the hypothalamus-pituitary-testicular axis (HPT axis) reduces free serum testosterone levels. In one embodiment, feedback on the hypothalamus-pituitary-testis axis (HPT axis) reduces the serum, tissue or tumor levels of androgens.

The hypothalamus-pituitary-testis axis (HPT axis) refers to an endocrine physiological system that regulates levels of hormone levels in the hypothalamus, pituitary gland, and testes. LHRH (luteinizing hormone releasing hormone) is released by the hypothalamus, which stimulates the pituitary to synthesize and secrete LH and FSH (gonadotropin). Next, LH and FSH act on the testicles to stimulate testosterone and sperm production. Next, testosterone has direct negative feedback on hypothalamic LHRH secretion and indirect negative feedback effects on pituitary LH and FSH production. Estrogens, androgens and serum proteins (e.g., inhivins) also have a negative effect on LHRH secretion and LH and FSH secretion.

The pituitary gland is a line that regulates the level of testosterone in the body. When testosterone levels are low, the pituitary releases luteinizing hormone (LH). This hormone causes the testicles to produce more testosterone. Testosterone levels increase during puberty. Testosterone levels are highest at approximately 20 to 40 years of age, and then gradually decrease in older men. Women have a much lower amount of testosterone in the body than men. However, testosterone plays an important role throughout the body in both men and women. This affects brain, bone and muscle mass, fat distribution, vascular system, energy levels, genital tissue, and sexual function. Most blood testosterone is bound to proteins called sex hormone binding globulin (SHBG) or to other serum proteins called albumin. Unbound (or "free") testosterone can also be clinically determined.

In another embodiment, a decrease in total serum testosterone or free serum testosterone levels independent of decreased serum luteinizing hormone levels is due to increased sex hormone-binding globulin (SHBG). In another embodiment, the decrease in total serum testosterone or free serum testosterone levels is due to an increase in sex hormone-binding globulin (SHBG). In another embodiment, a decrease in total serum testosterone or free serum testosterone levels independent of a decrease in serum luteinizing hormone (LH) level is due to inhibition of testosterone production and secretion by the testicular Rhodiol cells. In another embodiment, a decrease in total serum testosterone or free serum testosterone levels independent of decreased serum luteinizing hormone (LH) levels is due to decreased adrenocortical production.

In one embodiment, the compounds described herein and / or compositions comprising thereof can be used for the reduction of the level of luteinizing hormone (LH). In other embodiments, the compounds and / or compositions of the invention may be used for endogenous hormone reduction.

Hydroxysteroid dehydrogenase (HSD) classes are involved in the conversion of circulating steroids. 17 [beta] -HSD5 converts androstenedione to testosterone and estrone to estradiol. It is also accompanied by prostaglandin synthesis. In one embodiment, the compounds of the present invention inhibit HSD, especially 17 [beta] -hydroxysteroid dehydrogenase 5 (17 [beta] -HSD5). This inhibition can be used to prevent peripheral / extra-testosterone synthesis, which can lead to an incomplete decrease in total or free serum testosterone off the HPT axis control, or to allow for a locally increased intracellular testosterone level, all of which are deleterious to ADT , Which will be useful for ADT.

LHRH agonist therapy, that is, the androgen blockade regimen (ADT) achieved by administering a luteinizing hormone releasing hormone agonist (LHRH) or its analogue, initially stimulates testosterone production from the gonadotropin release from the pituitary gland and testes, reaction), then reduce gonadotrophin release and reduce both testosterone and estrogen levels. The "immediate response " caused by LHRH agonist therapy has a negative effect on prostate cancer treatment due to increased levels of androgen / testosterone. In addition, LHRH therapy is associated with an increased risk of diabetes and cardiovascular disease (Smith (2008) Current Prostate Reports . 6 : 149-154).

To overcome the immediate effects of LHRH therapy, anti-androgen monotherapy (bicalutamide, flutamide, chlormadinone), LHRH / antiandrogen combination therapy approach, and LHRH antagonist (degalelix) have been proposed (Suzuki et al, (2008) Int J. Clin Oncol 13:...... 401-410; Sharifi, N. et al, (2005) JAMA 294 (2): 238-244). Antiandrogen monotherapy does not reduce the level of androgen in the subject. Bicalutamide antiandrogen monotherapy appears to be less effective than ADT in prostate cancer patients with bone metastases. In addition, adverse effects observed in bicalutamide therapy include breast cancers, breast enlargement (gynecomastia and breast cancer) (Suzuki et al., Ibid). Additional risks of antiandrogen therapy include increased liver transaminase (Sharifi et al. Ibid).

In one embodiment, the present invention overcomes the side effects associated with estrogen deficiency caused by testosterone reduction using the traditional ADT method, without producing "flare" And / or reduced free serum testosterone levels. Use of the methods / compounds of the present invention may be beneficial for bone tissue maintenance (agonist effects on bone tissue), thrombotic potential and / or facial flushing and / or for less or neutral breast tissue than estradiol or diethylstilbestrol Lt; RTI ID = 0.0 &gt; effect-selective &lt; / RTI &gt; estrogenic activity.

In one embodiment, compound IV will not cause gonadotropin and testosterone increase, since compound IV exhibits agonist effects, but does not exhibit antagonist effects (Examples 6 and 7).

In one embodiment, compound IV exhibits agonist activity (Examples 8-11), demonstrating a strong pharmacological response to serum hormone, testosterone and total androgen reduction.

In one embodiment, the methods provided herein employing the compounds and / or compositions provided herein are effective in reducing or eliminating bone resorption caused by LH reduction using conventional ADT. In one embodiment, the methods provided herein and / or the use of compositions provided herein are effective in reducing or eliminating bone resorption caused by testosterone reduction using traditional ADT. In one embodiment, the methods provided herein using the compositions provided herein are effective in reducing or eliminating bone resorption caused by a decrease in estrogen as a result of LH level reduction. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein prevent bone resorption associated with reduced levels of LH using conventional ADT. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein prevent bone loss associated with endogenous LH, testosterone, and / or estradiol reduction using conventional ADT. In one embodiment, the methods provided herein using compounds and / or compositions provided herein increase bone density (BMD) while providing a reduction in LH levels. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein increase the bone volume percentage while providing endogenous LH, testosterone and / or estradiol levels.

In some embodiments, the invention provides a method of avoiding and / or reducing thromboembolism by administering a compound of the invention, an isomer thereof, a pharmaceutical product, a polymorph, a hydrate or a combination thereof.

In one embodiment, the methods provided herein using the compounds and / or compositions provided herein are effective for breast tissue. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein provide a reduction in LH levels while preventing gynecomastia associated with decreased LH levels achieved by conventional ADT.

In one embodiment, Example 13 discloses specific toxicity studies, wherein in vitro studies using human platelets show that compound IV has a much lower procoagulant activity than DES. Thus, the ER-selective agonist, Compound IV, will deliver the prostate cancer benefit of DES with less risk of thrombotic complications than DES, and will also deliver the benefits of LHRH agonists or antagonists without causing bone loss, flushing or harmful lipid profiles .

Diethylstilbestrol (DES) alone or in combination with other ADTs has shown that DES prevents bone resorption in patients with prostate cancer. Although the use of DES has been known as a treatment for prostate cancer, it is thought that the effects of DES on angiogenesis and malignant tumors are mediated by the DES metabolite and are not thought to act through estrogen receptors. Also, dosing levels of DES administered for therapeutic purposes suggest a number of adverse side effects including vascular disease, cardiovascular disease, thrombotic toxicity, gynecomastia, erectile dysfunction and decreased sexual desire (Scherr and Pitts, ibid and Presti, JC Jr. (1996) JAMA, 275 (15): 1153-6).

In one embodiment, the present invention overcomes the side effects of LHRH agonists or antagonist therapies, either alone or in combination with anti-androgen or DES. In another embodiment, the methods of the present invention provide androgen block therapy without estrogen-stimulating adverse effects, such as metaplastic breast, without adverse estrogen-deficient side effects such as harmful bone related symptoms. In another embodiment, the methods of the present invention overcome side effects associated with increased estrogen agonists commonly observed in DES therapy and overcome side effects associated with estrogen deficiency caused by LH reduction, And thus a reduction in total and / or free serum testosterone levels. The use of the methods / compounds of the present invention provides tissue-selective estrogenic activity to provide bone tissue retention (agonist effect on bone tissue), reduced thrombotic potential and a neutral effect on breast tissue.

The antiestrogen effect at the hypothalamic level of traditional selective estrogen receptor modulators (SERMs) such as tamoxifen, toremifene and raloxifene increases the level of gonadotropin or LH in men and potentially increases testosterone serum levels (Tsouri et al. al., 2008, Fertility and Sterility doi: 10.1016). In contrast, the methods of the present invention provide LH reduction in male subjects comprising administration of a compound of formula IA, I-XII

The addition of compounds of formula I Example :

In one embodiment of the method of the present invention, Y of the compound of formula I is C (O). In other embodiments, Y is CH _2. In other embodiments, R ₁ and R ₂ of the compounds of formula I or IA are independently O-Alk-NR ₅ R ₆ Or an O-Alk-heterocycle. In other embodiments, the O-Alk- _{heterocycle, O-Alk-NR 5 R} 6, -Alk- heterocycle and Alk-NR ₅ R ₆ Alk is a linear alkyl, 1-7 carbon of 1-7 carbon Branched alkyl, or cyclic alkyl of 3-8 carbons. In another embodiment, the alkyl is ethylene (-CH ₂ CH ₂ -). In other embodiments, the Alk is methylene (-CH ₂ -) a. In another embodiment, Alk is propylene (-CH ₂ CH ₂ CH ₂ -). In other embodiments, the Alk is 2-methyl-propylene _{(-CH 2 CH (CH 3)} CH 2 -) a.

이다. 본 발명의 방법의 다른 구현예에서, 식 I 또는 IA의 화합물의 R₁ 는 히드록실이다. 본 발명의 방법의 다른 구현예에서, 식 I 또는 IA의 화합물의 R₁ 는 알콕시이다. 본 발명의 방법의 다른 구현예에서, 식 I 또는 IA의 화합물의 R₁ 및 R₂ 는 독립적으로 수소, 할로겐, 히드록실, 알콕시, 시아노, 니트로, CF₃, N(R)₂, 술폰아미드, SO₂R, 알킬, 할로알킬, 아릴, O-Alk-NR₅R₆ 또는 O-Alk-헤테로사이클이며, 상기 헤테로사이클은 임의로 방향족인, 3-7 원 치환 또는 비치환 헤테로사이클 고리 이다. 본 발명의 방법의 다른 구현예에서, 식 I 또는 IA의 화합물의 R₁ 및 R₂ 는 독립적으로, 할로겐, 히드록실, 알콕시, 시아노, 니트로, CF₃, N(R)₂, 술폰아미드, SO₂R, 알킬, 할로알킬, 아릴, O-Alk-NR₅R₆ 또는 O-Alk-헤테로사이클이며, 상기 헤테로사이클은 임의로 방향족인, 3-7 원 치환 또는 비치환 헤테로사이클 고리 이다. 다른 구현예에서, 식 I 또는 IA의 화합물의 R₂ 는 할로겐이다. 다른 구현예에서, 식 I 또는 IA의 화합물의 R₂ 는 F이다. 다른 구현예에서, 식 I 또는 IA의 화합물의 R₂ 는 Cl이다. 다른 구현예에서, 식 I 또는 IA의 화합물의 R₂ 는 Br이다. 다른 구현예에서, 식 I 또는 IA의 화합물의 R₂ 는 I이다. 다른 구현예에서, 식 I 또는 IA의 화합물의 R₂ 는 히드록실이다. 다른 구현예에서, R₁ 및/또는 R₂ 는 CF₃이다. 다른 구현예에서, R₁ 및/또는 R₂ 는 CH₃ 이다. 다른 구현예에서, R₁ 및/또는 R₂ 는 할로겐이다. 다른 구현예에서, R₁ 및/또는 R₂ 는 F이다. 다른 구현예에서, R₁ 및/또는 R₂ 는 Cl이다. 다른 구현예에서, R₁ 및/또는 R₂ 는 Br이다. 다른 구현예에서, R₁ 및/또는 R₂ 는 I이다. 다른 구현예에서, 식 I의 R₂는 파라 포지션에 있다.In one embodiment of the method of the present invention, R ₁ of the compound of formula I or IA is in the para position. In one embodiment of the method of the invention, R ₁ and R ₂ of the compounds of formula I or IA are different. In another embodiment of the method of the present invention, R ₁ and R ₂ of the compound of formula I or IA are the same. In another embodiment of the method of the invention, R &lt; ₁ &gt; of the compound of formula I or IA is

to be. In another embodiment of the method of the present invention, R ₁ of the compound of formula I or IA is hydroxyl. In another embodiment of the method of the present invention, R ₁ of the compound of formula I or IA is alkoxy. In another embodiment of the method of the invention, formula (I) or R ₁ and R ₂ of the IA compound are independently hydrogen, halogen, hydroxyl, alkoxy, cyano, _{nitro, CF 3, N (R)} 2, sulfonamide , SO ₂ R, alkyl, haloalkyl, _{aryl, O-Alk-NR 5 R} 6 Or an O-Alk-heterocycle, said heterocycle being optionally aromatic, a 3-7 membered substituted or unsubstituted heterocycle ring. In another embodiment of the method of the invention, formula (I) or as R ₁ and R ₂ are independently of IA compound, halogen, hydroxyl, alkoxy, cyano, _{nitro, CF 3, N (R)} 2, sulfonamide, SO ₂ R, alkyl, haloalkyl, aryl, O-Alk-NR ₅ R ₆ Or an O-Alk-heterocycle, said heterocycle being optionally aromatic, a 3-7 membered substituted or unsubstituted heterocycle ring. In another embodiment, R ₂ of the compounds of formula I or IA is halogen. In another embodiment, R ₂ of the compound of formula I or IA is F. In another embodiment, R &lt; ₂ &gt; of the compound of formula I or IA is Cl. In other embodiments, R ₂ of the compounds of formula I or IA is Br. In another embodiment, R ₂ of the compound of formula I or IA is I. In another embodiment, R &lt; ₂ &gt; of the compound of formula I or IA is hydroxyl. In another embodiment, R ₁ and / or R ₂ is CF _3. In other embodiments, R ₁ and / or R ₂ is CH ₃ . In other embodiments, R ₁ and / or R ₂ is halogen. In another embodiment, R &lt; _{1 &gt;} and / or R &lt; ₂ &gt; In other embodiments, R &lt; _{1 &gt;} and / or R &lt; ₂ &gt; In other embodiments, R &lt; _{1 &gt;} and / or R &lt; ₂ &gt; In another embodiment, R &lt; _{1 &gt;} and / or R &lt; ₂ &gt; In another embodiment, R &lt; ₂ &gt; in formula I is in the para position.

In one embodiment of the method of the invention, R ₃ and R ₄ of the compound of formula I or IA are the same. In other embodiments, R ₃ and R ₄ of the compounds of formula I or IA are different. In other embodiments, j and k of the compounds of formula I or IA are independently 1. In another embodiment, R ₃ and R ₄ of the compounds of formula I or IA are independently halogen, haloalkyl, hydroxyl or alkyl. In other embodiments, R ₃ and R ₄ of the compounds of formula I or IA are independently F. In other embodiments, R ₃ and R ₄ of the compounds of formula I or IA are independently Br. In other embodiments, R ₃ and R ₄ of the compounds of formula I or IA are independently Cl. In another embodiment, R &lt; ₄ &gt; is in para position. In another embodiment, R &lt; ₃ &gt; is in the ortho position. In another embodiment, R &lt; ₃ &gt; is in a meta position. In other embodiments, R ₃ and / or R ₄ is CF ₃ . In other embodiments, R ₃ and / or R ₄ is CH ₃ .

In one embodiment of the method of the present invention, R ₅ and R ₆ of the compounds of formula I or IA, together with the nitrogen atom, form a 3-7 membered ring. In another embodiment, the ring is a saturated or unsaturated ring. In another embodiment, the ring is a substituted or unsubstituted ring. In other embodiments, R &lt; ₅ &gt; and R &lt; ₆ &gt; of the compounds of formula I or IA together with the nitrogen form a piperidine ring. In other embodiments, R &lt; ₅ &gt; and R &lt; ₆ &gt; of the compounds of formula I or IA together with the nitrogen form a pyrazine ring. R ₅ and R ₆ of the compounds of formula I or IA together with the nitrogen form a piperazine ring. R ₅ and R ₆ of the compounds of formula I or IA together with the nitrogen form a morpholine ring. R ₅ and R ₆ of the compounds of formula I or IA together with the nitrogen form a pyrrole ring. R &lt; ₅ &gt; and R &lt; ₆ &gt; of the compounds of formula I or IA form pyrrolidines. R ₅ and R ₆ of the compounds of formula I or IA together with the nitrogen form a pyridine ring. In another embodiment, the ring is substituted by halogen, alkyl, alkoxy, alkylene, hydroxyl, cyano, nitro, amino, amide, COOH, or aldehyde.

In another embodiment of the process of the invention, R ₁ of the compounds of the formula I or IA and R ₂ of the compounds of the formula I or IA are independently O-Alk-heterocycle or OCH ₂ CH ₂ -heterocycle. In another embodiment, the term "heterocycle" group means, in one embodiment, a ring structure that includes, in addition to carbon atoms, sulfur, oxygen, nitrogen or a combination thereof as part of a ring. In another embodiment, the heterocycle is a 3-12 membered ring. In another embodiment, the heterocycle is a six membered ring. In another embodiment, the heterocycle is a 5-7 membered ring. In another embodiment, the heterocycle is a 4-8 membered ring. In another embodiment, the heterocyclic is unsubstituted or halogen, haloalkyl, hydroxyl, alkoxy, carbonyl, amido, alkyl, amido, dialkyl amido, cyano, nitro, CO ₂ H, amino, alkylamino , Dialkylamino, carboxyl, thio and / or thioalkyl. In another embodiment, the heterocycle may be fused with another saturated or unsaturated cycloalkyl or heterocyclic 3-8 membered ring. In another embodiment, the heterocycle ring is a saturated ring. In another embodiment, the heterocycle ring is an unsaturated ring. In another embodiment, the heterocycle is piperidine. In another embodiment, the heterocycle is pyridine. In another embodiment, the heterocycle is piperidine, pyridine, furan, thiophene, pyrrole, pyrrolidine, pyrazine, piperazine or pyrimidine.

The term "cycloalkyl" means a non-aromatic, monocyclic or polycyclic ring containing carbon and hydrogen atoms. The cycloalkyl group has at least one carbon-carbon double bond in the ring, so long as the ring is not aromatic by the presence of a carbon-carbon double bond. Examples of the cycloalkyl group include, but are not limited to, a (C3-C7) cycloalkyl group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, and a cycloalkyl group such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl And saturated cyclic and bicyclic terpenes such as cycloheptenyl, and (C3-C7) cycloalkenyl groups, and unsaturated cyclic and bicyclic terpenes. The cycloalkyl group may be substituted or unsubstituted by one or more substituents. Preferably, the cycloalkyl group is a monocyclic ring or a bicyclic ring.

The term "alkyl" in one embodiment means a saturated aliphatic hydrocarbon comprising straight chain, branched chain and cyclic alkyl groups. In one embodiment, the alkyl group has 1-12 carbons. In another embodiment, the alkyl group has 1-7 carbons. In another embodiment, the alkyl group has 1-6 carbons. In another embodiment, the alkyl group has 1-4 carbons. In another embodiment, the cyclic alkyl group has 3-8 carbons. In another embodiment, the cyclic alkyl group has 3-12 carbons. In another embodiment, the branched alkyl is alkyl substituted by an alkyl side chain of 1-5 carbons. In other embodiments, the branched alkyl is alkyl substituted by a haloalkyl side chain of 1-5 carbons. Wherein said alkyl group is optionally substituted with one or more substituents selected from the group consisting of halogen, haloalkyl, hydroxyl, alkoxycarbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxyl, thio and / It can be unsubstituted.

"Alkenyl" in another embodiment means an unsaturated hydrocarbon comprising straight chain, branched chain and cyclic groups having at least one double bond. The alkenyl group may have one double bond, two double bonds, three double bonds and the like. In another embodiment, the alkenyl group has 2-12 carbons. In another embodiment, the alkenyl group has 2-6 carbons. In another embodiment, the alkenyl group has 2-4 carbons. Examples of alkenyl groups include ethenyl, propenyl, butenyl, cyclohexenyl and the like. The alkenyl group may be unsubstituted or substituted with halogen, hydroxy, alkoxycarbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxyl, thio and / or thioalkyl .

The term "aryl" group refers to a group selected from halogen, haloalkyl, hydroxy, alkoxy, carbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxy or thio or thioalkyl Means an aromatic group having at least one carbocyclic aromatic group or heterocyclic aromatic group which may be substituted or unsubstituted with one or more groups. Non-limiting examples of aryl rings are phenyl, naphthyl, pyranyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyrazolyl, pyridinyl, furanyl, thiophenyl, thiazolyl, imidazolyl, isoxazolyl, and the like . In one embodiment, the aryl group is a 4-8 membered ring. In other embodiments, the aryl group is a 4-12 membered ring. In another embodiment, the aryl group is a 6-membered ring. In another embodiment, the aryl group is a 5 membered ring. In another embodiment, the aryl group is a 2-4 fused ring system.

"Aldehyde" group refers to an alkyl or alkenyl group substituted in one embodiment by a formyl group, wherein the alkyl or alkenyl is as previously defined. In another embodiment, the aldehyde group is an aryl or phenyl group substituted with a formyl group, and the aryl is as defined above. Examples of aldehydes are formyl, acetal, propanal, butanal, pentanal, benzaldehyde. In another embodiment, the aldehyde group is a formyl group.

The "haloalkyl" group is, in another embodiment, an alkyl group as defined above substituted with one or more halogen atoms, such as F, Cl, Br,

The "hydroxyl" group is, in another embodiment, an OH group. It will be understood by those skilled in the art that when R &lt; ₁ &gt;, R &lt; _{2 &gt;} or R &lt; ₃ &gt; in the compounds of the present invention are OR, R is not OH.

In one embodiment, the term " halogen "or" halo " means a halogen such as F, Cl, Br or I.

In another embodiment, "phenol" refers to an alcohol (OH) derivative of benzene.

For protected hydroxyl groups, in some embodiments, incorporation of a substituent bonded to the oxygen moiety of the benzene ring, said substituent being readily removable. In some embodiments, the phenolic protecting group is selected from the group consisting of methyl ether (methoxy), alkyl ether (alkoxy), benzyl ether (Bn), methoxymethyl (MOM) ether, benzoyloxymethyl , Methoxyethoxymethyl (MEM) ether, 2- (trimethylsilyl) ethoxymethyl (SEM) ether, methylthiomethyl (MTM) ether, phenylthiomethyl (PTM) ether, azidomethyl ether, cyanomethyl ether (THF) ether, 1-ethoxyethyl (EE) ether, phenanthrene (THF) But are not limited to, ether, 4-bromophenacyl ether, cyclopropylmethyl ether, allyl ether, propargyl ether, isopropyl ether, cyclohexyl ether, t-butyl ether, 2,5-dimethylbenzyl ether, Ether, o-nitrobenzyl ether, 2,6-dichlorobenzyl ether, 3-4-dichlorobenzyl Tetralin, heptafluoro-o-tolyl, tetrafluoro-4-pyridyl, tetrafluoro-4-pyridyl, Butyldiphenylsilyl (TBDPS) ether, triisopropylsilyl (TIPS) ether, aryl formate, aryl acetate, aryl levulinate, , Aryl pivaloate, aryl benzoate, aryl 9-fluorenecarboxylate, arylmethyl carbonate, 1-adamantyl carbonate, t-butyl carbonate, t-methylsulfinylbenzyl carbonate, 2,4- 3-yl carbonate, aryl 2,2,2-trichloroethyl carbonate, aryl benzyl carbonate, aryl carbamate, dimethylphosphinyl ether (Dmp-OAr), dimethylphosphinothioyl ester (Mpt-OAr) (Dpt-OAr), aryl It may include a carbon-sulfonate, aryl pole toluene sulfonate or aryl 2-formyl-benzenesulfonate.

In one embodiment, the process of the present invention is a process for preparing N, N-bis (4-hydroxyphenyl) -4-propylbenzamide (II) or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate Or a combination thereof. In another embodiment, the process of the present invention is a process for the preparation of 4,4 '- (2,3-dimethyl-benzyllaurel) diphenol (III) or its isomers, pharmaceutically acceptable salts, pharmaceutical products, polymorphs, Hydrate or a combination thereof. In another embodiment, the process of the present invention is a process for the preparation of 3-fluoro-N- (4-fluorophenyl) -4-hydroxy-N- (4- hydroxyphenyl) benzamide Acceptable salts, pharmaceutical products, polymorphs, hydrates or combinations thereof. In another embodiment, the process of the present invention is a process for the preparation of N, N-bis (4-hydroxyphenyl) -2,3-dimethylbenzamide (V) or an isomer, pharmaceutically acceptable salt, , Hydrate, or a combination thereof. In another embodiment, the process of the present invention is a process for the preparation of N, N-bis (4-hydroxyphenyl) -2-naphthylamide (VI) or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate Or a combination thereof. In another embodiment, the process of the present invention is a process for the preparation of 3-fluoro-4-hydroxy-N, N-bis (4-hydroxyphenyl) benzamide VII or its isomers, Product, polymorph, hydrate or a combination thereof. In another embodiment, the process of the present invention is a process for the preparation of 4 - ((4-fluorophenyl) (4-hydroxybenzyl) amino) phenol (VIII) or an isomer, pharmaceutically acceptable salt, , Hydrate, or a combination thereof. In another embodiment, the process of the present invention is a process for the preparation of 4-fluoro-N- (4-hydroxy-phenyl) -N- [4- (2-piperidin- -Trifluoromethyl-benzamide (IX) or an isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or a combination thereof. In another embodiment, the process of the present invention is a process for the preparation of 4-fluoro-N- (4-hydroxy-phenyl) -N- [4- (2- (piperidin- (X) or an isomer, a pharmaceutically acceptable salt, a pharmaceutical product, a polymorph, a hydrate thereof, or a combination thereof. In an example, the process of the present invention is a process for the preparation of 3-fluoro-4-hydroxy-N- (4-hydroxyphenyl) -N-phenylbenzamide (XI) or an isomer, (XII), a polymorph, a hydrate, or a combination thereof. In another embodiment, the process of the present invention comprises reacting 3-fluoro-N, N-bis- (4-hydroxy- Or an isomer thereof, a pharmaceutically acceptable salt, a pharmaceutical product, a polymorph, a hydrate or a combination thereof.

In one embodiment, the methods of the present invention employ "pharmaceutically acceptable salts" of such compounds that can be produced by the reaction of an acid or base with a compound of the present invention.

Suitable pharmaceutically acceptable salts of the amines of the compounds of the process of the present invention may be prepared from inorganic or organic acids. In one embodiment, the inorganic salt of the amine is selected from the group consisting of bisulfate, borate, bromide, chloride, hemisulfate, hydrobromate, hydrochloride, 2-hydroxyethylsulfonate (hydroxyethanesulfonate), iodate, (Alkylsulfonates, arylsulfonates, halogen-substituted alkylsulfonates, halogen-substituted arylsulfonates), sulfonates, and thioesters, Cyanate.

In one embodiment, examples of organic salts of amines can be selected from aliphatic, alicyclic, aromatic, aromatic aliphatic, heterocyclic, carboxylic and sulfonic organic acids, such as acetate, arginine, aspartate, Alginate, benzenesulfonate, benzoate, bisulfate, butyrate, bicarbonate, butyrate, carboxylate, carboxylate, alkoxylate, alkoxycarbonate, But are not limited to, lactose, camphorate, camphorsulfonate, cyclohexyl sulfamate, cyclopentane propionate, calcium edetate, camylate, carbonate, clavulanate, cinnamate, dicarboxylate, digluconate, dodecylsulfonate, Dihydrochloride, decanoate, enanthuates, ethanesulfonate, But are not limited to, lactose, tartrate, eddylate, estolate, ethylate, fumarate, formate, fluoride, galacturonate, gluconate, glutamate, glycolate, glucoate, glucoheptanoate, glycerol phosphate, But are not limited to, glycyrrhizanate, glutaryl arsanylate, glutarate, glutamate, heptanoate, hexanoate, hydroxymaleate, hydroxycarboxylic acid, hexylresorcinate, hydroxybenzoate, hydroxynaphtholate, hydrofluorate, lactate , Maltodextrin, maltitol, maltodextrin, maltitol, maltodextrin, lactobionate, laurate, malate, maliate, methylenebis (beta-oxynaphtholate), malonate, mandelate, mesylate, methanesulfonate, methylbromide, Maleate, muate, monocarboxylate, nitrate, naphthalene sulfone Or a pharmaceutically acceptable salt thereof. The present invention also relates to a method for producing a compound of formula (I), wherein the compound is a compound selected from the group consisting of N, N'-dicyclohexylcarbodiimide, , Phthalates, pacinates, phenylpropionates, palmitates, pantothenates, polygalaturates, pyruvates, quinides, salicylates, succinates, stearates, sulfanilates, subacetates, , Theophylline acetate, p-toluenesulfonate (tosylate), trifluoroacetate, terephthalate, tanate, theoclate, trihaloacetate, triethiodide, tricarboxylate, undecanoate and valerate Rate.

In one embodiment, examples of inorganic salts of carboxylic acids or phenols include alkali metals, including ammonium, lithium, sodium, potassium, cesium; Alkaline earth metals including calcium, magnesium and aluminum; Zinc, barium, choline, and quaternary ammonium.

In other embodiments, examples of organic salts of carboxylic acids or phenols include organic amines including arginine, aliphatic organic amines, alicyclic organic amines, aromatic organic amines, benzanthene, t-butylamine, venetamine (N-benzylphenethylamine ), Dicyclohexylamine, dimethylamine, diethanolamine, ethanolamine, ethylenediamine, hydrabamine, imidazole, lysine, methylamine, meglamine, N-methyl-D-glucamine, N, But are not limited to, ethylenediamine, nicotinamide, organic amines, omithine, pyridine, picolies, piperazine, procaine, tris (hydroxymethyl) methylamine, triethylamine, triethanolamine, trimethylamine, tromethamine and urea Lt; / RTI &gt;

In one embodiment, the salt is prepared by reacting a product of the free base or free acid form with one or more appropriate acid or base equivalents in a solvent or medium insoluble in the salt or in a solvent such as water, Can be formed by conventional means, such as by vacuum or by lyophilization, or by exchanging ions of an existing salt with another ion or a suitable ion exchange resin.

In one embodiment, the methods of the present invention utilize pharmaceutically acceptable salts of the compounds of the present invention. In one embodiment, the methods of the present invention utilize a pharmaceutically acceptable salt of a compound of formula IA, I-XII. In one embodiment, the methods of the present invention utilize amine salts of compounds of formula IA, I-XII. In one embodiment, the process of the present invention utilizes a phenol salt of a compound of formula IA, I-XII.

In one embodiment, the process of the present invention comprises reacting a free base, free acid, non-charged or non-complexed compound of formula IA, I-XII, and / or an isomer, pharmaceutical product, hydrate, polymorph, Combination.

In some embodiments, the compounds of the invention comprise three phenyl groups attached by amide linkages. In one embodiment, the compounds of the present invention have a non-charged structure. In other embodiments, the compounds of the invention are free base structures. In other embodiments, the compounds of the invention are free acid structures. In other embodiments, the compounds of the invention are non-complexed structures. In other embodiments, the compounds of the invention are non-ionised structures. In other embodiments, the compounds of the present invention are pharmaceutically acceptable salts. In other embodiments, the compounds of the present invention comprise a hydrochloric acid (HCl) salt.

In one embodiment, the methods of the present invention utilize isomers of the compounds of Formulas IA, I-XII. In one embodiment, the method of the invention utilizes a pharmaceutical product of a compound of formula IA, I-XII. In one embodiment, the process of the present invention utilizes a hydrate of a compound of formula IA, I-XII. In one embodiment, the methods of the present invention utilize a polymorph of the compound of formula IA, I-XII. In one embodiment, the methods of the present invention utilize metabolites of compounds of formula IA, I-XII. In another embodiment, the methods of the present invention utilize a composition comprising a compound of formula IA, I-XII, or, in another embodiment, an isomer, metabolite, metabolite of a compound of formula IA, I-XII, , Hydrates, and polymorphs.

In one embodiment, the term "isomer" includes, but is not limited to, optical isomers and analogs, structural isomers and analogs, morphologic isomers and analogs, and the like.

In one embodiment, the term "isomer" is meant to include the optical isomer of the compound. In one embodiment, the term "isomer" is meant to include stereoisomers of such compounds. The compounds of the present invention have amide bonds which may be in the cis or trans isomerization thereof. It is to be understood that the invention encompasses optically active, or stereoisomeric forms, or mixtures thereof, and uses thereof for applications considered within the scope of the present invention.

In another embodiment, the present invention includes a hydrate of the compound. In one embodiment, the term "hydrate" includes the hemihydrate, monohydrate, dihydrate, trihydrate, and the like as is known in the art.

Synthesis procedure

Compounds of formula I or IA can be readily prepared, for example, by reacting a substituted diphenylamine with benzoic acid or a benzoyl halide in the presence of a base to produce the benzamide. In one embodiment, the base is pyridine. In another embodiment, the benzoyl halide is benzoyl chloride. In another embodiment, the hydroxyl substituent is protected during the reaction of diphenylamine with benzoic acid or benzoyl chloride. In another embodiment, the protecting group for the hydroxyl is optionally removed in the final step. See U.S. Pat. No. 2009/00624231, which is incorporated herein by reference in its entirety.

For example, compounds of formula IA:

Wherein R ₁ , R ₂ , R ₃ and R ₄ , j and k are as defined above;

을

of

와 반응시켜

Gt;

을 생산하고;

&Lt; / RTI &gt;

The diphenylamine (3) is reacted in the presence of a base

와 반응시켜

Gt;

And a step of reacting the compound of formula

Wherein _{R, R 1, R 2, R} 3 and R ₄ are independently _{OH, O-Alk-R 5} R 6 or is O-Alk- _{heterocycle, R 1 ', R 2'} , R 3 ', R 4 'is a protected hydroxyl group, to obtain the free hydroxyl group by removing the protecting group or optionally to obtain a compound of Cl-Alk- or heterocyclyl Cl-Alk-NR ₅ R ₆ to yield formula IA:

When R ₁ , R ₂ , R ₃ and R ₄ are independently other than OH, O-Alk-NR ₅ R ₆ or O-Alk-heterocycle, R ₁ ', R ₂ ', R ₃ 'and R ₄ 'are R ₁ , R ₂ , R ₃ and R _4, respectively.

As another example, the process for preparing a compound of formula IA comprises:

(Wherein R ₁ , R ₂ , R ₃ and R ₄ are as defined above)

In the presence of a base

을

of

와 반응시켜

Gt;

. &Lt; / RTI &gt;

Wherein _{R, R 1, R 2, R} 3 and R ₄ are independently _{OH, O-Alk-R 5} R 6 or is O-Alk- _{heterocycle, R 1 ', R 2'} , R 3 ', R 4 'is a protected hydroxyl group, to obtain the free hydroxyl group by removing the protecting group or optionally to obtain a compound of Cl-Alk- or heterocyclyl Cl-Alk-NR ₅ R ₆ to yield formula IA:

When R ₁ , R ₂ , R ₃ and R ₄ are independently other than OH, O-Alk-NR ₅ R ₆ or O-Alk-heterocycle, R ₁ ', R ₂ ', R ₃ 'and R ₄ 'are R ₁ , R ₂ , R ₃ and R _4, respectively.

In one embodiment, Formula II is prepared according to Example 1 and FIG.

In another embodiment, Formula III is prepared according to Example 1 and FIG.

In a further embodiment, the compound of formula IV is:

In the presence of a base

을

of

와 반응시켜

Gt;

을 수득한 후,

, &Lt; / RTI &gt;

The protecting group is deprotected to give compound IV:

를

To

&Lt; / RTI &gt;

(Wherein P and P 'are the same or different protecting groups). In one embodiment, compound IV is prepared according to Example 2 and FIG.

In another embodiment, compound V is prepared according to Example 1 and FIG.

In a further embodiment, compound VI is prepared according to Example 3 and FIG.

In another embodiment, compound VII is prepared according to Example 1 and FIG.

In another embodiment, Compound VIII is prepared according to Example 4 and FIG.

In another embodiment, compound IX is prepared according to Example 5 and FIG.

In another embodiment, Compound X hydrochloride is prepared according to Example 5 and FIG.

In another embodiment, compound XI is prepared according to Example 1 and FIG.

In another embodiment, compound XII is prepared according to example 1 and figure 5.

Suitable hydroxyl protecting groups include, for example, methyl ether (methoxy), benzyl ether (benzyloxy) methoxymethyl (MOM) ether, benzoyloxymethyl (BOM) ether, benzyl, carbobenzoxy, methoxyethoxymethyl (Methyl) ether, methylthiomethyl (MTM) ether, phenylthiomethyl (PTM) ether, azidomethyl ether, cyanomethyl ether, 2,2-dichloro Ethoxyethyl (EE) ether, phenacyl ether, 4-bromoethyl ether, tetrahydrofuranyl ether, tetrahydrofuranyl ether, Propyl ether, isopropyl ether, cyclohexyl ether, t-butyl ether, benzyl ether, 2,6-dimethylbenzyl ether, 4-methoxybenzyl ether, o - nitrobenzyl ether, 2,6-dichlorobenzyl ether, 3,4-dichlorobenzyl 4-pyridyl, heptafluoro-p-tolyl, tetrafluoro-4-pyridyl, 4-methylphenyl Butyldiphenylsilyl (TBDPS) ether, triisopropylsilyl (TIPS) ether, aryl formate, aryl acetate, aryl levulinate, , Aryl pivaloate, aryl benzoate, aryl 9-fluorenecarboxylate, arylmethyl carbonate, 1-adamantyl carbonate, t-butyl carbonate, 4-methylsulfinylbenzyl carbonate, 2,4- 3-yl carbonate, aryl 2,2,2-trichloroethyl carbonate, aryl benzyl carbonate, aryl carbamate, dimethyl phosphinyl ester (Dmp-OAr), dimethyl phosphinothioyl ester (Mpt-OAr) (Dpt-OAr), &lt; / RTI &gt; And a methanesulfonate, aryl toluenesulfonate or aryl 2-formyl-benzenesulfonate.

The methods of the invention include the use of compounds IA, I-XII wherein the preparation of the compounds of the present invention comprises reacting diphenylamine with benzoyl chloride in the presence of a base. Suitable bases include, for example, pyridine, _{triethylamine, K 2 CO 3, Cs 2} CO 3, Na 2 CO 3, methylamine, imidazole, benzimidazole, histidine, tributylamine, or a combination thereof. In one embodiment, the base is pyridine.

The methods of the present invention include the use of compounds IA, I-XII, wherein the preparation of the compounds of the present invention involves deprotection of the protected hydroxyl. In another embodiment, the deprotection conditions are according to a protecting group. In some embodiments, the deprotection step comprises hydrogenation in the presence of Pd / C. In another embodiment, the deprotection comprises reaction with BBr &lt; _{3 &} gt ;. In another embodiment, the deprotecting step comprises reacting with an acid.

In a further embodiment, compounds IA, I-XII are prepared according to Figs. 5-8 and 1-5.

Pharmaceutical composition

In some embodiments, the invention includes administering a composition comprising the compound. The term "pharmaceutical composition" as used herein means a "therapeutically effective amount" of an active ingredient, i.e., a compound of the invention, and a pharmaceutically acceptable carrier or diluent. "Therapeutically effective amount" means an amount that provides a therapeutic effect for a given condition and method of administration.

As used herein, the term "administering " means contacting a subject with a compound of the invention. Administration as used herein can be accomplished in vitro, i. E. In a test tube, or in vivo, i. E. In an organism, e. In one embodiment, the invention includes administering a compound of the invention to a male subject.

In another embodiment, the invention includes a pharmaceutical product of a compound described herein. "Pharmaceutical product" means, in another embodiment, a composition (pharmaceutical composition) suitable for pharmaceutical use as described herein.

The compounds of the present invention may be administered alone or as an active ingredient of a preparation. Thus, the present invention also encompasses pharmaceutical compositions of the compounds of formula I, including, for example, one or more pharmaceutically acceptable carriers.

A number of standard references describing procedures for preparing various agents suitable for administration of the compounds according to the present invention are useful. Examples of possible preparations are, for example, the Handbook of Pharmaceutical Excipients, American Pharmaceutical Association (current edition); Are included in Pharmaceutical Dosage Forms: Tablets (Lieberman, Lachman and Schwartz, editors) current edition, published by Marcel Dekker, Inc., and Remington's Pharmaceutical Sciences (Arthur Osol, editor), 1553-1593 (current edition).

The mode of administration and formulation are closely related to the amount of treatment of a compound or composition that is desirable and effective for a given therapeutic application.

Suitable formulations include, but are not limited to, oral, rectal, sublingual, mucosal, nasal, eye, subcutaneous, intramuscular, intravenous, transdermal, spinal, intrasynovial, intraarticular, subarachnoid, , Intrauterine administration, and formulations for systemic delivery of other active ingredients. Formulations suitable for oral administration are preferred.

For the preparation of such pharmaceutical formulations, the active ingredient is mixed with the pharmaceutical carrier according to typical pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration.

In preparing the compositions of the oral formulations, any of the usual pharmaceutical media may be employed. Thus, for liquid oral formulations such as, for example, suspensions, elixirs and solutions, suitable carriers and additives include water, glycols, oils, alcohols, flavoring agents, preservatives, colorants and the like. For solid oral preparations such as, for example, powders, capsules and tablets, suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like. Because of their ease of administration, tablets and capsules represent the most advantageous oral unit dosage forms. If desired, tablets may be sugar-coated or enteric coated by standard techniques.

For parenteral formulations, the carrier will usually comprise sterile water and may contain other ingredients, for example ingredients for assisting in solubility or for preservation. Injectable solutions may also be prepared using suitable stabilizers.

In some applications, the active agent is covalently bound, chelated or coordinated to suitable biomolecules such as encapsulation in liposomes or other encapsulation media or the active agent, for example, from proteins, lipoproteins, glycoproteins and polysaccharides Quot; vectorized "form, such as &lt; / RTI &gt;

The therapeutic methods of the present invention employing agents suitable for oral administration may each be presented as discrete units such as capsules, cachets, tablets or lozenges, containing a predetermined amount of the active ingredient, for example, as a powder or granules have. Optionally, suspensions in aqueous liquid or non-aqueous liquids, such as syrups, elixirs, emulsions or drafts, may be used.

Tablets may optionally be made by compression or molding, or wet granulation, with one or more accessory ingredients. Compressed tablets may be prepared by compressing the active compounds in a suitable machine, for example, in the form of free-flowing, such as powders or granules optionally mixed with binders, disintegrants, lubricants, inert diluents, surfactants or dragees. Molded tablets comprising a powdered active compound and a suitable carrier can be prepared by molding in a suitable machine.

Syrups may be prepared by adding the active compound to a concentrated aqueous solution of sugar, e. G., Sucrose, and also adding thereto any auxiliary component (s). Such ancillary component (s) may include flavoring agents, suitable preservatives, agents that delay the crystallization of sugars, and agents that increase the solubility of polyhydroxy alcohols, such as glycerol or sorbitol, for example.

Formulations suitable for parenteral administration include sterile aqueous preparations of the active compound, which are preferably isotonic with the blood of the recipient (e. G., Physiological saline). Such agents may include suspending agents and thickening agents and liposomes or microparticulate systems designed to target the compounds to a blood component or one or more organs. The formulations may be presented in unit dosage or multiple dosage forms.

Parenteral administration may include any suitable form of systemic delivery. Administration may be by, for example, intravenous, intraarterial, intrathecal, intramuscular, subcutaneous, intraperitoneal, and the like, and may be carried out by means suitable for an infusion pump (external or implantable) or other desired mode of administration.

Nasal and other mucosal spray preparations (e. G., Inhalable forms) may contain the active ingredient and a purified aqueous solution of preservatives and isotonic agents. Such a preparation is preferably adjusted to a pH and an isotonic state compatible with the nose or other mucous membranes. Alternatively, it may be in the form of a solid powder finely divided in a gas carrier. Such formulations may be delivered by any suitable means or method, for example, a nebulizer, atomizer, metered dose inhaler, and the like.

Formulations for rectal administration may be presented as a suppository with a suitable carrier such as cocoa butter, hydrogenated fats or hydrogenated fats carboxylic acid.

The dermal preparation may be prepared by incorporating the active agent into a thixotropic or gelatinous carrier such as a cellulose medium, for example methylcellulose or hydroxyethylcellulose, and then dissolving the resulting formulation in a transdermal device adapted to be fixed in skin contact with the skin of the wearer &Lt; / RTI &gt;

In addition to the above ingredients, the formulations of the present invention may contain one or more accessory ingredient (s) selected from, for example, diluents, buffers, flavoring agents, binders, disintegrants, surfactants, thickeners, lubricants, preservatives May be further included.

The formulations of the present invention may have immediate release, sustained release, delayed release or other release profiles known in the art.

In one embodiment, the invention provides a method of treating a subject suffering from prostate cancer, comprising administering an oral composition comprising a compound of Formula IA, I-XII, A) decreased total serum testosterone levels; b) providing a method of reducing free serum testosterone levels. In an additional embodiment, the methods of the invention comprise administering an oral composition comprising a compound of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, Formula X, Formula XI or Formula XII .

In one embodiment, the invention provides a method of treating a subject suffering from prostate cancer, comprising administering an oral composition comprising a compound of Formula IA, I-XII, Provides a way to treat prostate cancer indifferently. In an additional embodiment, the invention provides a method of administering an oral composition comprising a compound of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, Formula X, Formula XI or Formula XII The present invention provides a method of treating prostate cancer, either by LH reduction or in relation to LH hormone reduction in a male subject having prostate cancer.

It will be appreciated that the present invention encompasses any embodiment of the compounds described herein as "compounds of the invention" in some embodiments.

In one embodiment, the methods of the invention comprise administering a compound of the invention in varying dosages. In one embodiment, the compound of the invention is administered at a dose of 1-1500 mg per day. In an additional embodiment, the compounds of the invention are administered at a dose of 1-10 mg, 3-26 mg / day, 3-60 mg / day, 3-16 mg / day, 3-30 mg / day, 10-26 mg / day, 15 -60 mg, 50-100 mg per day, 50-200 mg per day, 150-300 mg per day, 25-50 mg per day, 5-50 mg per day, 200-500 mg per day, 150-500 mg per day, 200-1000 mg per day , 300-1500 mg per day, or 100-1000 mg per day.

In one embodiment, the methods of the invention comprise administering a compound of the invention in varying dosages. In one embodiment, the compound of the invention is administered in a dose of 3 mg. In additional embodiments, the compounds of the invention are administered in dosages of 10 mg, 30 mg, 50 mg, 100 mg, 200 mg, 300 mg, 450 mg, 500 mg, 600 mg, 900 mg, 1000 mg or 1500 mg.

In one embodiment, the methods of the invention comprise administering a compound of the invention in varying dosages. In one embodiment, a compound of the invention is administered at a dose of 0.1 mg / kg / day. In additional embodiments, the compounds of the invention may be administered at a dose of 0.2 to 30 mg / kg / day, or 0.2 mg / kg / day, 0.3 mg / kg / day, 1 mg / kg / mg / kg / day, 10 mg / kg / day, 20 mg / kg / day, or 30 mg / kg / day.

In one embodiment, the methods of the invention provide for the use of a pharmaceutical composition comprising a compound of formula IA, I-XII. In an additional embodiment, the methods of the present invention comprise administering a pharmaceutical composition comprising a compound of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, Formula X, Formula XI or Formula XII The use of the composition is provided.

In certain embodiments, the pharmaceutical composition is a solid dosage form. In another embodiment, the pharmaceutical composition is a tablet. In another embodiment, the pharmaceutical composition is a capsule. In another embodiment, the pharmaceutical composition is a solution. In another embodiment, the pharmaceutical composition is a transdermal patch.

In one embodiment, a compound of the invention or a composition comprising it will have the use of inhibiting, inhibiting, promoting or stimulating a desired response in a subject, as will be understood by those skilled in the art. In other embodiments, the composition will additionally comprise additional active ingredients having a useful activity for the particular application to which the compounds of the present invention are administered.

For administration to mammals, particularly humans, the physician will determine the actual dosage and duration of treatment, which will be best suited to the individual and will vary with age, weight, sex, and / or the response of a particular individual.

In some embodiments, the compositions of the present invention will comprise the compounds of the invention in any form or embodiment described herein. In some embodiments, the compositions of the invention will consist of a compound of the invention in any form or embodiment described herein. In some embodiments, the compositions of the invention will consist essentially of the compounds of the invention in any form or embodiment described herein. In some embodiments, the term "comprising" encompasses the indicated active ingredients and other active agents, such as the compounds of the invention, and pharmaceutically acceptable carriers, excipients, emollients, stabilizers, and the like, it means. In some embodiments, the term "essentially consisting" is intended to encompass the case where the indicated active ingredient is the sole active ingredient, or other compounds that are not intended to stabilize, preserve, or otherwise directly affect the therapeutic effect of the indicated active ingredient &Lt; / RTI &gt; In some embodiments, the term "essentially consisting" may refer to a component that promotes the release of the active ingredient. In some embodiments, the term "consisting" means a composition containing the active ingredient and a pharmaceutically acceptable carrier or excipient.

Any use of any of the compounds described herein may be used in the treatment of any disease, disorder, or condition described herein, and will be understood to represent embodiments of the invention. In one embodiment, the compound is a free base, a free acid, a non-charged or non-complexed compound.

The following examples are presented to more fully illustrate preferred embodiments of the invention. However, these should not be construed as limiting the scope of the invention in any way.

Example

Example  One

General synthetic procedures and synthetic intermediates for compounds of formulas II-XII

Organic solvents, surfactants, and antioxidants and the like that can be used in the compositions described herein are typically readily available from commercial sources. For example, PEG-300, polysorbate 80, Captex ^TM 200, Capmul ^TM MCM C8 can be purchased from Dow Chemical Company (Midland, MI), ICI Americas, Inc. (Wilmington, DE) or Abitec Corporation .

The estrogen receptor ligands described herein can be prepared in a variety of ways known in the art. For example, the estrogen receptor ligands described herein can be prepared by the synthetic methods described in U.S. Patent No. 2009/0062341, the disclosure of which is incorporated herein by reference in its entirety.

N, N - Bis  General Synthesis of Aryl Benzamide Derivatives

In general synthesis of diaryl aniline (5). A mixture of arylamine (1.5 eq.), Aryl iodide (1 eq.), K ₂ CO ₃ (2 eq.), CuI (0.1 eq.) And L-proline (0.2 eq.) Were mixed together and dissolved in anhydrous DMSO Respectively. The reaction mixture was then stirred and heated at 90 &lt; 0 &gt; C for 28 hours. The mixture was cooled to room temperature and hydrolyzed with water. The solution was partitioned by addition of EtOAc. The EtOAc layer separated, washed with brine, dried over anhydrous MgSO _4. The solvent was removed under reduced pressure. The solid residue was purified by flash column chromatography (silica gel) using 5% EtOAc / hexane as eluent to give the corresponding diaryl aniline.

Bis- (4 -methoxyphenyl ) amine (1a) : pale yellow solid, 73% yield. Mp 98.6-99.0 [deg.] C. _{^{1 H NMR (CDCl 3, 300}} MHz) δ6.93-6.81 (m, 8H), 5.37 (s, br, 1H), 3.78 (s, 6H). MS m / z 228.4 (MH) ^&lt; ⁺ & gt ^;

N- (4 -methoxyphenyl ) phenylamine (1b) : pale yellow solid, 70% yield. Mp 106.3-106.5 [deg.] C. ¹ H NMR (CDCl ₃ , 300 MHz)? 7.24-7.18 (m, 3H), 7.08-7.06 (m, 2H), 6.92-6.84 s, 3H). MS m / z 200.1 (M + H) &lt; ⁺ & gt ^; .

N- (4- Fluorophenyl ) -N-4 -methoxyphenylamine (1c) : pale yellow solid, 54% yield. Mp 60.6-61.0 [deg.] C. _{^{1 H NMR (CDCl 3, 300}} MHz) δ7.01-6.83 (m, 8H), 3.78 (s, 3H). MS m / z 217 (M) ^&lt; ⁺ & gt ^; .

N- (4- benzyloxyphenyl ) -N-4 -methoxyphenylamine (1d) : pale yellow solid, 54% yield. Mp 108.0-108.4 [deg.] C. _{^{1 H NMR (CDCl 3, 300}} MHz) δ7.34-7.08 (m, 5H), 6.90-6.81 (s, 3H), 3.78 (s, 3H). MS m / z 306 (M + H) &lt; ⁺ & gt ^; .

General synthesis of benzamide . A mixture of aryl aniline (1 eq), benzoyl chloride (1.3 eq), and pyridine (6 eq) were mixed together and dissolved in anhydrous THF at room temperature. The mixture was stirred and refluxed for 24 hours. The reaction solution was cooled to room temperature and hydrolyzed by addition of 2N HCl solution. The solution was extracted with ethyl acetate. Washing the organic layer with saturated NaHCO ₃ solution to remove excess acid, dried over anhydrous MgSO _4, and concentrated under reduced pressure. The residue was purified by flash column chromatography using EtOAc / hexane (3/7 v / v) to give the corresponding benzamide compound.

3- Fluoro- N- (4- fluorophenyl ) -4 -methoxy- N- (4 -methoxyphenyl ) benzamide (2a) : yellow solid, Mp 54-56 [deg.] C. _{^{1 H NMR (CDCl 3 / TMS}} ) δ 7.24-7.11 (m, 4H), 7.05-6.97 (m, 4H), 6.85-6.78 (m, 3H), 3.86 (s, 3H), 3.79 (s, 3H) . MS (ESI) m / z 370.1 [M + H] &lt; ⁺ &gt;

4- Fluoro - N, N - bis (4 -methoxyphenyl ) -2- ( trifluoromethyl ) benzamide (2b) : colorless oil, 84.2% yield. _{^{1 H NMR (CDCl 3, 300}} MHz) δ7.34-7.26 (m, 4H), 7.09-7.01 (m, 3H), 6.91 (d, 2H, J = 8.7 Hz), 6.87 (d, 2H, J = 8.7 Hz), 3.80 (s, 3H), 3.71 (s, 3H). MS m / z 442.1 (M + Na) &lt; ⁺ & gt ^; .

4-methoxy -N- (4-methoxyphenyl) -N- (4-fluoro-phenyl) -benzamide (2c): white solid, 97% ^{yield, Mp 133.5-134.5 ℃ 1 H NMR (} CDCl 3, 300 MHz) [delta] 8.11-6.66 (m, 15H), 3.74 (s, 3H), 3.73 (s, 3H). MS m / z 384 (M + H) &lt; ⁺ & gt ^; .

N- (4 -methoxyphenyl ) -N- (4- benzoylphenyl ) -2 -naphthylamide (2d) : white solid, 58% yield. Mp 174.9-175.5 [deg.] C. ¹ H NMR (CDCl ₃ , 300 MHz)? 8.04 (s, 1H), 7.77-7.74 (m, 2H), 7.64-7.61 (m, 1H), 7.51-7.43 (m, 4H), 7.40-7.31 m, 4H), 7.13-7.10 m, 4H), 6.88-6.78 (m, 4H), 4.99 (s, 2H), 3.74 (s, 3H). MS m / z 460 (M + H) &lt; ⁺ & gt ^; .

4- Fluoro - N, N - bis (4 -methoxyphenyl ) -2- ( trifluoromethyl ) benzamide (2e) : colorless oil, 84.2% yield. _{^{1 H NMR (CDCl 3, 300}} MHz) δ7.34-7.26 (m, 4H), 7.09-7.01 (m, 3H), 6.91 (d, 2H, J = 8.7 Hz), 6.87 (d, 2H, J = 8.7 Hz), 3.80 (s, 3H), 3.71 (s, 3H). MS m / z 442.1 (M + Na) &lt; ⁺ & gt ^; .

General procedure for the demethylation of a benzamide derivative using BBr _3. The methoxybenzamide compound was dissolved in dry CH ₂ Cl ₂ . BBr ₃ (1.0 M CH ₂ Cl ₂ solution) was added dropwise at 0 ° C. The reaction solution was slowly warmed to room temperature and stirred overnight at room temperature. The mixture was cooled to 0 &lt; 0 &gt; C in an ice bath and hydrolyzed by the addition of water. The solution was partitioned by addition of EtOAc. Separating the organic layer; The aqueous layer was extracted with EtOAc. The organic layer was washed with brine and dried over anhydrous MgSO _4. The solvent was removed under reduced pressure. The residue was purified by flash column chromatography using CH ₃ OH / CH ₂ Cl ₂ (1/9 v / v) to give the corresponding phenolic compound.

4- Fluoro - N, N - bis (4 -hydroxyphenyl ) -2- ( trifluoromethyl ) benzamide (3a): white solid, 92.5% yield. _{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ9.55 (s, 1H), 9.53 (s, 1H), 7.69-7.58 (m, 2H), 7.46-7.39 (m, 1H), 7.18 (d, 2H, J = 8.7 Hz), 6.93 (d, 4H, J = 8.7Hz), 7.03 (d, 2H, J = 8.4 Hz), 6.78 (d, 2H, J = 8.7 Hz), 6.57 (d, 2H, J = 8.7 Hz). MS m / z 392.1 (M + H) &lt; ⁺ & gt ^; .

The following compounds were synthesized and characterized as described hereinabove and summarized in Table 1: N, N-bis (4-hydroxyphenyl) -4-propylbenzamide (II); 3-fluoro-N- (4-fluorophenyl) -4-hydroxy-N- (4-hydroxyphenyl) benzamide (IV); N, N-bis (4-hydroxyphenyl) -2,3-dimethylbenzamide (V); 3-Fluoro-4-hydroxy-N, N-bis (4-hydroxyphenyl) -benzamide (VII); 3-fluoro-4-hydroxy-N- (4-hydroxyphenyl) -N-phenylbenzamide (XI); And 3-fluoro-N, N-bis (4-hydroxyphenyl) -2-methylbenzamide (XII).

General procedure for debenzylation of benzyloxyphenyl -benzamide . The compound was dissolved in EtOH in a 250 mL hydrogenation vessel. Pd / C powder (5% mol) was added to the solution. The reaction vessel was placed in a hydrogenation unit under 20 psi pressure hydrogen gas. The reaction was monitored until the starting material disappeared. Then, the solvent was removed under reduced pressure. The residue was purified by flash column chromatography using hexane / EtOAc = 3/2 v / v to give the desired product.

The following compounds were synthesized and identified as described herein and summarized in Table 1: N, N-bis (4-hydroxyphenyl) -2-naphthylamide (VI).

General procedure for the reduction of deprotected benzamide . The benzamide compound was dissolved in 20 mL dry THF at room temperature. H ₃ B (SMe ₂ ) was added at room temperature via argon under vacuum. The reaction solution was stirred and heated to reflux for 6 hours. The reaction was then quenched by adding 10 mL of MeOH at 0 &lt; 0 &gt; C. The solvent was removed under reduced pressure. The residue was subjected to flash column chromatography (silica gel, CH ₂ Cl ₂ / MeOH = 9/1 v / v) to obtain the desired product.

The following compounds were synthesized and identified as described herein and are summarized in Table 1: 4,4 '- (2,3-Dimethylbenzylla turyl) diphenol (III); 4 - ((4-fluorophenyl) (4-hydroxybenzyl) amino) phenol (VIII).

General synthesis of O- (2-piperidin-1 -ylethoxy) -benzamide and analogs . To the hydroxyphenyl solution containing the benzamide analog (1 eq) in acetone was added K ₂ CO ₃ (3 eq) and N-chloroethyl-piperidine hydrochloride (1.2 eq.). The solution was heated to reflux for 6 hours. The solution was evaporated to dryness. Water was added to hydrolyze the residue and then extracted with ethyl acetate. The organic layer was separated and dried over anhydrous MgSO _4. The solvent was removed under reduced pressure. The residue was purified by flash chromatography using methylene chloride / methanol = 9/1 v / v to give the desired compound. HCl in Et ₂ O was added to a methanol solution of the above compound, and then the solvent was evaporated to prepare a hydrochloride.

The following compounds were synthesized and identified as described herein and are summarized in Table 1: 4-Fluoro-N- (4-hydroxyphenyl) -N- (4- (2-piperidin- Ethoxy) phenyl) -2- (trifluoromethyl) benzamide (IX); And 4-fluoro-N- (4-hydroxyphenyl) -N- (4- (2- (piperidin- 1 -yl) ethoxy) phenyl) -2- (trifluoromethyl) ) Benzamide hydrochloride (X).

Compound # rescue Physical identification II

^{1 H NMR (DMSO-d6,} 300 MHz) δ 9.46 (s, 2H, 2 X OH), 7.27-7.26 (m, 2H, ArH), 7.06-7.04 (m, 2H, ArH), 6.99-6.97 (m 2H, CH2), 0.82 (t, J = 7.33 (m, 2H, CH2), 4H, ArH), 6.66-6.65 Hz, 3H, CH3). m / z 346.0 (MH) - III

Tan foam, 41% yield. Mp 147-150 [deg.] C. _{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ 8.92 (s, 2H), 7.07 (d, J = 7.33 Hz, 1H), 7.00-6.94 (m, 2H), 6.76-6.72 (m, 4H), 4H), 4.72 (s, 2H), 2.23 (s, 3H), 2.16 (s, 3H). m / z 320.2 (M + H) &lt; ⁺ & gt ^; IV

Tan solid, 92% yield. Mp 110-112 [deg.] C. _{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ10.14 (bs, 1H), 9.71 (bs, 1H), 7.26-7.11 (m, 5H), 7.05-6.99 (m, 3H), 6.78 (t, J = 8.61 Hz, 2H), 6.68 (d, J = 8.68 Hz, 2H). m / z 364.1 (M + Na) &lt; ⁺ & gt ^; V

^{1 H NMR (DMSO-d6,} 300 MHz) δ9.47 (bs, 2H, 2 X OH), 7.18 (d, J = 8.30 Hz, 2H, ArH), 7.06 (d, J = 7.08 Hz, 1H, ArH 2H, ArH), 2.22 (s, 3H, CH3), 7.00-6.92 (m, 4H, ArH), 6.78 (d, J = 8.30 Hz, 2.15 (s, 3H, CH3). m / z 334.3 (M + H) &lt; + &gt; VI

White solid, 70% yield. Mp 264.3-265.2 C (decomposed). _{^{1 H NMR (DMSO- d 6,}} 500 MHz) δ9.46 (s, 2H), 7.98 (s, 1H), 7.85-7.75 (m, 2H), 7.75-7.73 (m, 2H), 7.54-7.48 ( m, 2H), 7.45-7.43 (m, 1H), 7.05 (s, 4H), 6.66 (s, 4H). m / z 356 (M + H) &lt; ⁺ & gt ^; VII

_{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ10.25 (bs, 1H, OH), 9.48 (bs, 2H, 2 X OH), 7.12-6.95 (m, 6H, ArH), 6.80-6.65 (m , 5H, ArH). m / z 338.0 (MH &lt; ⁺ &gt;) ^- VIII

Yellow oil, 92% yield. _{^{1 H NMR (DMSO- d 6,}} 500 MHz) δ9.29 (s, 1H), 9.24 (s, 1H), 7.09 (d, 2H, J = 8.3 Hz), 6.98 (d, 2H, J = 9.0 Hz ), 6.94-6.91 (m, 2H), 6.73 (d, 2H, J = 9.0 Hz), 6.68-6.64 (m, 4H), 4.70 (s, 2H). m / z 307.8 (MH) ^- IX
And X
(HCl salt of IX)

White solid, 57.7% yield. _{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ9.57 (s, 1H), 7.71-7.68 (m, 2H), 7.47-7.44 (m, 1H), 7.28 (d, 1H, J = 9.0 Hz) , 7.18 (d, 1H, J = 8.7 Hz), 7.13 (d, 1H, J = 8.7 Hz), 7.05 (d, 1H, J = 8.4 Hz), 6.97 (d, 1H, J = 9.0 Hz), 6.80 -6.76 (m, 2H), 6.57 (d, 1H, J = 87. Hz), 4.06 (t, 1H, J = 6.0 Hz), 3.93 (t, 1H, J = 6.0 Hz), 2.66 (t, 1H 2H, J = 5.7 Hz), 2.55 (t, 1H, J = 5.4 Hz), 2.44 (s, 2H), 2.36 (s, 2H), 1.49-1.37 (m, 6H). m / z 501.0 (MH) ^- XI

^{1 H NMR (DMSO-d6,} 300 MHz) δ9.95 (bs, 1H, OH), 9.47 (bs, 2H, 2 X OH), 7.02-6.95 (m, 6H, ArH), 6.75-6.72 (m, 1H, ArH), 6.68-6.66 (m, 4H, ArH). m / z 324.0 (M + H) &lt; + &gt; XII

Light red solid. 72.0% Yield. Mp > _{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ9.50 (bs, 2H), 7.19-6.79 (m, 7H), 6.61 (d, J = 8.93 Hz, 2H), 6.53 (d, J = 7.79 Hz , &Lt; / RTI &gt; 2H), 2.23 (s, 3H). m / z 336.0 (MH &lt; ⁺ &gt;).

Example 2

Synthesis of Compound of Formula IV (Figure 6)

Step 1: Synthesis of 4-fluoro-N- (4-methoxyphenyl) aniline (1c)

Anhydrous K ₂ CO ₃ (122.23 g, 0.884 mol), CuI (11.23 g, 58.96 mmol), and L (4-fluoroaniline, 78.00 g, 0.708 mol) -Propyne (13.58 g, 0.118 mol) were mixed together in a dry 1 L 3-neck round bottom flask equipped with stir bar, reflux condenser and argon inlet. Anhydrous DMSO (300 mL) was added at room temperature. The reaction mixture was stirred and heated at 90 &lt; 0 &gt; C for 20 h under argon. The mixture was then cooled to room temperature and hydrolyzed with water (300 mL). The solution was partitioned by the addition of EtOAc (200 mL). The EtOAc layer was separated. The aqueous layer was extracted with 100 mL of EtOAc. The combination of the EtOAc layers, wash with brine (2 x 100mL) and dried over anhydrous MgSO ₄ (50 g). The solvent was removed under reduced pressure. The brown oil residue was purified by flash column chromatography (silica gel, hexane / EtOAc = 9/1 v / v) to give 4-fluoro- N- (4- methoxyphenyl) aniline (1c) 99.70 g, 77.6%. Mp 46-48 ° C. MS (ESI) m / z 218.1 [M + H] ⁺ , ¹ H NMR (DMSO- d ₆ , 300 MHz)? 7.77 (bs, 1H), 7.03-6.98 (m, 4H), 6.93-6.82 , &Lt; / RTI &gt; 4H), 3.70 (s, 3H).

step 2: 3 - Fluoro -N- (4- Fluorophenyl )-4- Methoxy -N- (4- Methoxyphenyl ) &Lt; / RTI &gt; benzamide (2a).

Fluoro-N- (4-methoxyphenyl) aniline (1c) (90.78 g, 0.418 mol) and 3-fluoro (3-fluoroaniline) were dissolved in a dry 1 L three-necked round bottom flask equipped with a stir bar, a reflux condenser and an argon inlet. -4-methoxybenzoyl chloride (94.55 g, 0.501 mol) were mixed together and dissolved in anhydrous THF (200 mL). Anhydrous pyridine (132.22 g, 1.672 mol) was added via syringe at room temperature under argon. The reaction mixture was stirred and heated to reflux overnight. The reaction mixture was cooled to room temperature and filtered to remove the pyridine salt. The solution was concentrated to remove the THF solvent. The residue oil was washed with 200 mL of 2N HCl solution and extracted with ethyl acetate (2 x 200 mL). The combined organic layers were washed with a saturated aqueous Na ₂ CO ₃ solution (150 mL) to remove excess benzoyl chloride and acid, dried over MgSO ₄ (50 g), and concentrated under reduced pressure to provide an oil. The residue was purified by flash column chromatography using silica gel with CH ₂ Cl ₂ / acetone (50/1 v / v) to give the pure corresponding benzamide compound as a yellow solid. Mp 54-56 [deg.] C. MS (ESI) m / z 370.1 [M + H] +, 1 H NMR (CDCl 3 / TMS) δ7.24-7.11 (m, 4H), 7.05-6.97 (m, 4H), 6.85-6.78 (m, 3H), 3.86 (s, 3H), 3.79 (s, 3H).

step 3: 3 - Fluoro -N- (4- Fluorophenyl ) -4-hydroxy-N- (4- Hydroxyphenyl ) &Lt; / RTI &gt; benzamide (IV).

A compound 3-fluoro -N- (4-fluorophenyl) -4-methoxy -N- (4-methoxyphenyl) benzamide (2a) (138.0 g, 0.374 mol) under argon and the drying at room temperature, CH ₂ Cl ₂ was dissolved in (600 mL). With stirring via a syringe at 0 ℃ in an ice bath under argon was added dropwise _{BBr 3 (374.75 g, 1.496 mol} ). The reaction solution was allowed to stir overnight at room temperature. Next, the solution was poured with 1 L of ice water while stirring. The slurry mixture was stirred at room temperature for 2 hours. The white precipitate was filtered, washed with water (2 x 100 mL) and dried under vacuum. The CH ₂ Cl ₂ layer was separated, dried over anhydrous MgSO ₄ (50 g), filtered, and concentrated to dryness under reduced pressure. The white precipitate and residue from the CH ₂ Cl ₂ solution were combined and purified by flash column chromatography (silica gel, CH ₂ Cl ₂ / acetone / MeOH = 90/7/3 v / v / v) to give a pale tan solid , Which was recrystallized twice from a hot EtOAc / hexane solution to give 104.0 g of a white crystalline solid in 81.6% yield. Mp 110-112 [deg.] C. MS (ESI) m / z 364.1 [M + Na] +, 1 H NMR (DMSO- d 6) δ10.14 (bs, 1H), 9.71 (bs, 1H), 7.25-7.11 (m, 5H), 7.05 6.99 (m, 3H), 6.78 (t, J = 8.6 Hz, 1H), 6.68 (d, J = 8.7 Hz, 2H).

Example 3

Synthesis of Compound of Formula VI (Figure 7)

Synthesis of 4- (benzoyloxy) -N- (4-methoxyphenyl) aniline (1d).

4-benzoyloxyaniline (16.6 g, 83.31 mmol), 4-iodoanisole (15.0 g, 64.09 mmol), K ₂ CO ₃ (17.72 g, 128.18 mmol), CuI (1.48 g, 12.82 mmol) were mixed together and dissolved in anhydrous DMSO (120 mL) at room temperature. The reaction mixture was then stirred and heated to 90 &lt; 0 &gt; C for 48 hours. The mixture was cooled to room temperature and hydrolyzed with water. The solution was partitioned by addition of EtOAc. The EtOAc layer separated, washed with brine, dried over anhydrous MgSO _4. The solvent was removed under reduced pressure. The solid residue was purified by flash column chromatography (silica gel) using EtOAc / hexanes (1/9 v / v) to give the corresponding diaryl aniline as a yellow solid in 9.8 g, 50% yield. Mp 108.0-108.4 [deg.] C. ¹ H NMR (CDCl ₃ , 300 MHz)? 7.34-7.25 (m, 5H), 6.90-6.81 (m, 8H), 5.02 (s, 2H), 3.78 (s, 3H). MS m / z 306 (M + H) &lt; ⁺ & gt ^; .

Synthesis of N- (4-benzoyloxyphenyl) -N- (4-methoxyphenyl) -2-naphthamide (2d).

(Benzoyloxy) -N- (4-methoxyphenyl) aniline (0.80 g, 2.62 mmol) was added to a 1.5 eq. Solution of 2- Naphthoyl chloride (0.75 g, 3.93 mmol) and 4 equivalents of pyridine (0.83 g, 10.48 mmol). The mixture was dissolved in anhydrous THF (30 mL) and heated to reflux for 20 h. The reaction solution was cooled to room temperature and filtered. The solvent was removed under reduced pressure. The residue was purified by flash column chromatography with silica gel with EtOAc / hexane (3/7 v / v) to give the pure corresponding naphtamide compound as a white solid in 0.70 g, 58% yield. Mp 174.9-175.5 [deg.] C. ¹ H NMR (CDCl ₃ , 300 MHz)? 8.04 (s, 1H), 7.77-7.74 (m, 2H), 7.64-7.61 (m, 1H), 7.51-7.43 (m, 4H), 7.40-7.31 m, 4H), 7.13-7.10 (m, 4H), 6.88-6.78 (m, 4H), 4.99 (s, 2H), 3.74 (s, 3H). MS m / z 460 (M + H) &lt; ⁺ & gt ^; .

Synthesis of N, N-bis (4-hydroxyphenyl) -2-naphthylamide (VI).

The compound N- (4-benzoyloxyphenyl) -N- (4-methoxyphenyl) -2-naphthamide (2d) (0.50 g, 1.09 mmol) was dissolved in dry CH ₂ Cl ₂ (30 mL) at room temperature . BBr ₃ (3.26 mL of 1.0 M CH ₂ Cl ₂ solution, 3.26 mmol) was added dropwise via syringe at room temperature with stirring. The reaction solution was allowed to stir overnight at room temperature. The mixture was cooled to 0 &lt; 0 &gt; C in an ice bath and water was added to hydrolyze. The solution was partitioned by addition of EtOAc. Separating the organic layer; The aqueous layer was extracted twice with EtOAc. Combine the organic layers, washed with brine, dried over anhydrous MgSO _4. The solvent was removed under reduced pressure. The residue was purified by flash column chromatography using silica gel with CH ₃ OH / CH ₂ Cl ₂ (1/9 v / v) to give the desired phenol compound as a white solid in 0.27 g, 70% yield. Mp 264.3-265.2 C (decomposed). _{^{1 H NMR (DMSO- d 6,}} 500 MHz) δ9.46 (s, 2H), 7.98 (s, 1H), 7.85-7.75 (m, 2H), 7.75-7.73 (m, 2H), 7.54-7.48 ( m, 2H), 7.45-7.43 (m, 1H), 7.05 (s, 4H), 6.66 (s, 4H). MS m / z 356 (M + H) &lt; ⁺ & gt ^; .

Example 4

Synthesis of compounds of formula VIII.

Synthesis of 4 - ((4-fluorophenyl) (4-hydroxybenzyl) amino) phenol (VIII)

The compound N- (4-fluorophenyl) -4-hydroxy-N- (hydroxyphenyl) benzamide (0.30 g, 0.93 mmol) was dissolved in 20 mL dry THF at room temperature. . H ₃ B (SMe ₂ ) (1.86 mL of 2 M THF solution, 3.71 mmol) was added via syringe at room temperature under argon. The reaction solution was stirred and heated to reflux for 6 hours. The reaction was then quenched by adding 10 mL of MeOH at 0 &lt; 0 &gt; C. The solvent was removed under reduced pressure. The residue was subjected to flash column chromatography (silica gel, CH ₂ Cl ₂ / MeOH = 9/1 v / v) to obtain 0.26 g of yellow oil in 92% yield. _{^{1 H NMR (DMSO- d 6,}} 500 MHz) δ9.29 (s, 1H), 9.24 (s, 1H), 7.09 (d, 2H, J = 8.3 Hz), 6.98 (d, 2H, J = 9.0 Hz ), 6.94-6.91 (m, 2H), 6.73 (d, 2H, J = 9.0 Hz), 6.68-6.64 (m, 4H), 4.70 (s, 2H). MS m / z 307.8 (MH) ^- .

Example 5

Synthesis of Compounds of Formula IX and X (Figure 8).

Synthesis of diaryl aniline . (1.5 eq), aryl iodide (1 eq), K ₂ CO ₃ (2 eq), CuI (1 eq) and L-proline (0.2 eq) were mixed together and dissolved in anhydrous DMSO at room temperature . The reaction mixture was then stirred and heated to 90 &lt; 0 &gt; C for 28 hours. The mixture was cooled to room temperature and hydrolyzed with water. The solution was partitioned by addition of EtOAc. The EtOAc layer separated, washed with brine, dried over anhydrous MgSO _4. The solvent was removed under reduced pressure. The solid residue was purified by flash column chromatography using EtOAc / hexane (3/7 v / v) as solvent to give the corresponding diaryl aniline. Bis- (4 -methoxyphenyl ) amine (1a) : pale yellow solid, 73% yield. _{^{1 H NMR (CDCl 3, 300}} MHz) δ6.93-6.81 (m, 8H), 5.37 (s, br, 1H), 3.78 (s, 6H). MS m / z 228.4 (MH) ^&lt; ⁺ & gt ^; .

4 flow - N, N - bis (4-methoxyphenyl) Synthesis of 2- (trifluoromethyl) benzamide (2e).

In a dry 3-neck round bottom flask equipped with a magnetic stirrer and a reflux condenser, one equivalent of bis- (4-butoxyphenyl) amine (1a) (0.73 g, 3.18 mmol) was added to a solution of 1.2 equivalent of 4-fluoro- -Trifluoromethylbenzoyl chloride (0.87 g, 3.82 mmol) and 6 equivalents of pyridine (1.51 g, 19.08 mmol). The mixture was dissolved in anhydrous THF (20 mL) and heated to 90 &lt; 0 &gt; C for 20 h. The reaction solution was cooled to room temperature and filtered. The solvent was removed under reduced pressure. The residue was purified by flash column chromatography with silica gel with EtOAc / hexane (3/7 v / v) to give pure corresponding benzamide compound as a colorless oil in 1.12 g, 84.2% yield. _{^{1 H NMR (CDCl 3, 300}} MHz) δ7.34-7.26 (m, 4H), 7.09-7.01 (m, 3H), 6.91 (d, 2H, J = 8.7 Hz), 6.87 (d, 2H, J = 8.7 Hz), 3.80 (s, 3H), 3.71 (s, 3H). MS m / z 442.1 (M + Na) &lt; ⁺ & gt ^; .

4- Fluoro - N, N - Bis (4- Hydroxyphenyl )-2-( Trifluoromethyl ) Benzamide (3a).

To a solution of 4-fluoro-N, N-bis (4-methoxyphenyl) -2- (trifluoromethyl) benzamide 2e (1.00 g, 2.38 mmol) in dry CH ₂ Cl ₂ ). BBr ₃ (10 mL of a 1.0 M CH ₂ Cl ₂ solution, 10.0 mmol) was added dropwise at room temperature via a syringe. The reaction solution was stirred overnight at room temperature. The mixture was cooled to 0 &lt; 0 &gt; C in an ice bath and water was added to hydrolyze. The solution was partitioned by addition of EtOAc. Separating the organic layer; The aqueous layer was extracted twice with EtOAc. The organic layer was combined, washed with brine, dry MgSO ₄ Lt; / RTI &gt; The solvent was removed under reduced pressure. The residue was purified by flash column chromatography using silica gel with CH ₃ OH / CH ₂ Cl ₂ (1/9 v / v) to give pure desired phenol compound as a white solid in yield of 0.86 g, 92.5%. _{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ9.55 (s, 1H), 9.53 (s, 1H), 7.69-7.58 (m, 2H), 7.46-7.39 (m, 1H), 7.18 (d, 2H, J = 8.7 Hz), 6.93 (d, 4H, J = 8.7Hz), 7.03 (d, 2H, J = 8.4 Hz), 6.78 (d, 2H, J = 8.7 Hz), 6.57 (d, 2H, J = 8.7 Hz). MS m / z 392.1 (M + H) &lt; ⁺ & gt ^; .

4- Fluoro -N- (4- Hydroxyphenyl ) -N- [4- (2-piperidin-1-yl) - Ethoxy ) Phenyl] -2- ( Trifluoromethyl ) &Lt; / RTI &gt; benzamide (IX)

To a solution of 4-fluoro-N, N-bis (4-hydroxyphenyl) -2 (trifluoromethyl) benzamide 3a (0.61 g, 1.56 mmol) in acetone was added K ₂ CO ₃ , 9.36 mmol) and N-chloroethyl-piperidine hydrochloride (0.34 g, 1.87 mmol). The solution was heated to reflux for 20 h. The solution was evaporated to dryness. The residue was purified by flash chromatography (silica gel; methylene chloride / methanol = 9/1 v / v) to give the desired compound as a white solid in 0.45 g, 57.7% yield. _{^{1 H NMR (DMSO- d 6,}} 300 MHz) δ9.57 (s, 1H), 7.71-7.68 (m, 2H), 7.47-7.44 (m, 1H), 7.28 (d, 1H, J = 9.0 Hz) , 7.18 (d, 1H, J = 8.7 Hz), 7.13 (d, 1H, J = 8.7 Hz), 7.05 (d, 1H, J = 8.4 Hz), 6.97 (d, 1H, J = 9.0 Hz), 6.80 -6.76 (m, 2H), 6.57 (d, 1H, J = 87. Hz), 4.06 (t, 1H, J = 6.0 Hz), 3.93 (t, 1H, J = 6.0 Hz), 2.66 (t, 1H 2H, J = 5.7 Hz), 2.55 (t, 1H, J = 5.4 Hz), 2.44 (s, 2H), 2.36 (s, 2H), 1.49-1.37 (m, 6H). MS m / z 501.0 (MH) ^- .

HCl in Et ₂ O was added to the methanol solution of the above compound, and the solvent was evaporated to prepare the hydrochloride (X).

Example 6

Estrogen receptor binding affinity, agonist and antagonist activity

(GST) fusion ER-alpha or ER-beta ligand binding domain ([ ³ H] E2) expressed by natural high affinity ER ligand [2,4,6,7- ³ H The binding affinity of the compounds was determined using competitive in vitro radioligand binding assays using the LBD protein.

Way

The equilibrium dissociation constant ( Kd ) of [ ³ H] E ₂ was determined by combining recombinant ER-α or ER-β with [ ³ H] E ₂ . Total and non-specific binding to measure, it was incubated with the protein with high concentrations of unlabelled E concentration of ^[3 H] E ₂ to increase in the presence or absence of ₂ during the 4 ℃ 18 hours. Except for the non-specific binding, the Kd of E ₂ (ERα: 0.71 nM; ERβ: 1.13 nM) was determined using nonlinear regression. In addition, [ ³ H] E ₂ required for saturation of ER-α and ER-βLBD was determined to be 4-6 nM.

Increasing concentrations of compound (range: 10 ^-11 to 10 ^-6 M) were incubated with [ ³ H] E ₂ (5.7 nM) and ER LBD using the above conditions. After incubation, the plates were harvested using a GF / B filter on a Unifilter-96 Harvester (PerkinElmer) and washed three times with very cold buffer B (50 mM Tris, pH 7.2). The filter plate was dried at room temperature and then 35 μl Microscint-O cocktail was added to each well and the filter plate was sealed with TopSeal-A. The radioactivity was counted in a TopCount® NXT Microplate Scintillation Counter using the setting for 3H in a Microscint cocktail (PerkinElmer).

^[3 H] (determined by incubating with 10 ^-6 M unlabeled E ₂₎ determining the specific binding of ^[3 H] E ₂ by subtracting the non-specific binding of E ₂ at each concentration of compound, and this compound under test member As a percentage of specific binding. The concentration of the compound (IC ₅₀ ) which reduced the specific binding of [ ³ H] E ₂ by 50% was determined. Next, the equilibrium binding constants ( Ki ) of the compounds were calculated as follows: K _i = K _d X IC ₅₀ / ( K _d + L), where K _d is the equilibrium dissociation constant of [ ³ H] E ₂ -α = 0.71 nM; ER-β = 1.13 nM), L is a ^[3 H] concentrations of _{E 2 (ER-α: 5.7} nM; ER-β: 5.7 nM).

result

Binding assays indicate that the ligand binds to ER- [alpha] and ER- [beta] with varying concentrations ranging from 3.75 nM to greater than 1000 nM and a non-isoform selective selectivity range in isoform selective. The results from representative compounds are listed in Table 2.

Binding results for selected compounds compound ER-a Ki (nM) ER-β Ki (nM)

3.75


81.6

3.81


6.44

21.7


15.2

7.13


35.9

9


72

6.06


76.92

13


19

14.79


646.32

15


57

15.12


25.02

Compound IV binds to ER [alpha] and ER [beta]. Of [2,4,6,7- ³ H (N)] of natural high-affinity ligand ER-estradiol ^([3 H] E ₂₎ and by the bacteria expressing GST-fusion that ERα or ERβ ligand binding domain (LBD) protein Lt; / RTI &gt; binding affinity of compound IV was determined using an in vitro competitive radioligand binding assay. In this assay, the ERa and ER [beta] binding affinity of Compound IV ( Ki Values) were 21.7 ± 1.7 nM (n = 3) and 15.2 ± 4.1 nM (n = 3), respectively. Upon binding to the ER, Compound IV initiates a series of complex molecular reactions that result in the expression or inhibition of a target gene involved in a pharmacological reaction in a tissue-selective manner. In transient transfection assays, compound IV is an ERa and ERbeta agonist and has been demonstrated to have greater ability to stimulate ERa-mediated transcriptional activity compared to ERbeta. Estradiol stimulates ERα and ERβ with a 5.1-fold greater selectivity for ERα, whereas Compound IV shows a 49.0-fold selectivity for ERα. Thus, compound IV has a relative 9.7-fold selectivity (normalized to the estradiol value) in relative transcriptional activity to ERa relative to ERβ. In addition, no antagonistic effect was observed in the estradiol (1 nM) -stimulatory transcriptional activity by Compound IV at a concentration of 10 [mu] M or less. While many steroid ligands cross-react with other nuclear hormone receptors, the action of compound IV is specific for ERa and ERb. Compound IV was tested for transcriptional activity in both the agonist and antagonist modes by the glucocorticoid receptor (GR), the mineralocorticoid receptor (MR), the progesterone receptor (PR), the rat isoform of the androgen receptor (AR) Screening for the cross reactivity of the receptor (FXR), liver X receptor (LXT), peroxisome proliferator activated receptor (PPAR- alpha and PPAR-y), and retinoit X receptor (RXR- Respectively. Compound IV did not show any agonist or antagonist activity in this assay, which supports the conclusion that compound IV does not functionally cross-react with these nuclear hormone receptor superfamily members.

Example  7

Transcriptional activation of selected compounds

Agonist and antagonist transcriptional activation assays were performed to ascertain whether the compound was an agonist, antagonist or partial.

Way

Rat estrogen receptors (ER-alpha and ER-beta) were cloned from rat ovarian cDNA into the pCR 3.1 plasmid vector backbone. Sequencing was performed to determine the mutation member. HEK-293 cells were plated at 100,000 cells per well of a 24 well plate in Dulbecco ' s Minimal Essential Media (DMEM) + 5% charcoal-removed fetal calf serum (csFBS). The cells were transfected with lipofectamine ((Invitrogen, Carlsbad, CA) with 0.25 ug of ERE-LUC, 0.02 ug CMV-LUC (renilla luciferase) and 12.5 ng of rat ER- The cells were treated with various concentrations of compounds or combinations of compounds and estradiol for 24 hours to determine antagonistic activity. After 48 hours of transfection, luciferase assays were performed.

result

Screening of the compounds of the invention in transcriptional activation systems has shown that the compounds of the present invention belong to all three classes of agonists, antagonists and partial agonists. Examples of agonists and antagonists are given in Table 3. The transcription activation results matched very well with the binding results for isoform selectivity.

Table 3 provides EC ₅₀ and IC ₅₀ transcriptional activation for some selected compounds of the invention.

compound ER-alpha
EC ₅₀  (nM) ER-β
EC ₅₀  (nM) ER-alpha
IC ₅₀  (nM) ER-β
IC ₅₀  (nM)

0.65




40.4



> 1000



> 1000

> 1000


> 1000


2.207


145

Example 8

Testosterone inhibition in cynomolgus monkeys

During the study according to the USDA guidelines (except fasting prior to oral administration), gonad-innoculated male cynomolgus monkeys (n = 2) at 2 years of age were admitted with free access to primate diets and water. Animals were dosed once daily with an oral gavage dose of 30 mg / kg of the compound of formula IV in a microemulsion Beihek of Tween 80 / deionized water for 7 consecutive days. At 1 (baseline), 3, 4, 5, 6, and 7 days, serum samples were withdrawn by venipuncture prior to oral dosing. Testosterone and total androgens were quantified using enzyme immunoassay (EIA) methods, both with or without the HPLC method. After 6 days of treatment of the compound of formula IV, a time-dependent reduction in testosterone and total androgen was evident (testosterone / dihydrotestosterone). Compounds of formula IV inhibited testosterone levels to 58% and 64%, respectively, in animals # 1 and # 3 compared to baseline values (solid line in Figure 1; see Table 4). Similarly, total androgen levels were 56% inhibited in animals # 1 and # 3 compared to baseline values (dashed line in Figure 1; see Table 4).

Consistent with the estrogen feedback in the pituitary-testis axis in males, these results indicate that serum hormones (testosterone and total) in the intact non-human primate (cynomolgus monkey) after repeated oral administration (30 mg / &Lt; / RTI &gt; androgen).

Testosterone and total androgen levels in uninfected male monkey serum given at a dose of 30 mg / kg of compound IV daily (first dose on Day 0). Testosterone Total androgen Serum levels (pg / mL) Serum levels (pg / mL) Work Animal 1 Animal 3 Animal 1 Animal 3 0 (basal value) 1120 617 1868 1643 2 937 479 1178 847 3 784 437 1078 786 4 552 415 988 924 5 403 276 966 664 6 474 221 819 726
Percent reduction from baseline
Percent reduction from baseline Work Animal 1 Animal 3 Animal 1 Animal 3 0 (basal value) 100 100 100 100 2 16 22 37 48 3 30 29 42 52 4 51 33 47 44 5 64 55 48 60 6 58 64 56 56

Example 9

Inhibition of LH and testosterone levels in rats

In vivo dose-response studies were performed to evaluate the effect of compound IV on LH inhibition in intact and testosterone (ORX) male rats. In intact and ORX animals, compound IV significantly inhibited LH levels at doses of &gt; 10 mg / kg daily compared to the respective controls. (Suppression of the same pattern was observed at the level of FSH). LH inhibition significantly reduced testosterone levels below the limit of 0.08 ng / mL (BLOQ), and the prostate, seminal vesicle and anal muscle mass were very androgen - dependent, reducing their weight. In intact animals, a dose-dependent reduction in these target organ crabs was noted for the seminal vesicle and anal ankle weights at the level of the fermented control group. Prostate weight was significantly reduced in uninjured animals, but these values did not reach the level of the sterile control group. The results are summarized in Table 6 below.

Materials and Methods :

Male Sprague-Dawley rats weighing approximately 200 g were maintained in a 12-h human / cancer cycle with an unlimited supply of food (2016 Teklad Global 16% Protein Rodent Diet, Harlan, Madison, Wis.) And water. The animal protocol was reviewed and approved by the Animal Experimental Ethics Committee of the University of Tennessee.

Test materials for this study were quantitated and dissolved in 10% DMSO (Fisher) diluted with PEG 300 (Acros Organics, NJ) to prepare suitable dosage formulations. For this study, 60 male Sprague-Dawley rats were randomly selected for body weight and designated as one of the 12 treatment groups (n = 5 animals / group). The treatment groups are listed in Table 5. The animals were housed in groups of 2-3 animals per group. Controls (intact and testicular resection (ORX)) were administered daily with Beckle. Compound IV was administered intradermally at doses of 0.3, 1, 3, 10 and 30 mg / kg / day and subcutaneously injected (200 μL) in all ORX groups.

After the 14 day dosing regimen, animals were sacrificed by anesthesia (ketamine / xylazine, 87:13 mg / kg) and body weight recorded. In addition, the abdominal prostate, seminal vesicle and anal muscle mass were removed, external tissue was removed, and quantified individually. Tissue weights were normalized to body weight and expressed as a percentage of uninjured control. Blood was collected from the abdominal aorta under isoflurane anesthesia and allowed to solidify. Serum was separated by centrifugation and stored at -80 ° C prior to measurement of serum hormone levels. Serum luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentrations were measured by rat pituitary lumenx assay (Millipore, Billerica, MA) according to manufacturer's instructions. The lower limit for this assay was 3.2 pg / mL for LH and 32 pg / mL for FSH. Testosterone was measured by testosterone EIA (Alpco Diagnostics, Salem, NH) with an LLOQ of 0.08 ng / mL. Serum hormone levels below the lower limit of quantification (BLOQ) were excluded from the group mean analysis. Therefore, the values reported for LH and T in the group with sample BLOQ are set to be less than the actual value. These analytical methods provided estimates for the most conservative LH and T inhibition. The individual treatment groups were compared to the uninjured and ORX Beihikle controls using the Fisher's Least Signigicant Difference test. The significance was presupposed to be P-value &lt; 0.05.

Treated group group Gonadal status Dose (mg / kg / day) Test substance One Intact - Beihik 2 ORX - Beihik 3 Intact 0.3 Compound IV 4 Intact One Compound IV 5 Intact 3 Compound IV 6 Intact 10 Compound IV 7 Intact 30 Compound IV 8 ORX 0.3 Compound IV 9 ORX One Compound IV 10 ORX 3 Compound IV 11 ORX 10 Compound IV 12 ORX 30 Compound IV

No damage and luteinizing hormone levels in ORX rats (Table 6)

The LH levels (mean ± SD) in the intact and ORX Beihoku controls were 1.46 ± 0.64 and 11.1 ± 3.9 ng / mL, respectively. Compound IV decreased LH levels in a dose-dependent manner in intact animals, reaching a statistically significant reduction of the daily dose ≥ 3 mg / kg. LH levels in untreated compound IV treated animals were 0.863 ± 0.384, 0.704 ± 0.530, 0.395 ± 0.302, 0.226 ± 0.165, and 0.236 ± 0.176 ng after administration of 0.3, 1, 3, 10 and 30 mg / / mL. LH level in ORX male was also significantly decreased by treatment with Compound IV. Within the ORX animals, LH levels were 15.4 ± 2.9, 13.5 ± 2.2, 6.5 ± 5.6, 0.425 ± 0.135, and 0.368 ± 0.119 ng / mL after administration of 0.1, 1, 3, 10 and 30 mg / Respectively. The results are shown graphically in Fig. 10a.

No damage and follicle stimulating hormone levels in ORX rats (Table 6)

FSH levels in the uninjured and ORX Beihik controls were 20.9 ± 8.5 and 93.5 ± 13.8 ng / mL, respectively. In intact animals, Compound IV decreased FSH levels dose-dependently and a significant decrease was observed at dose ≥ 10 mg / kg / day. FSH levels in untreated Compound IV treated animals were 17.3 ± 6.4, 15.7 ± 7.3, 18.4 ± 7.7, 9.2 ± 4.0, and 6.3 ± 1.8 ng, respectively, after administration of 0.3, 1, 3, 10 and 30 mg / / mL. In ORX animals, LH levels were 115 ± 17, 114 ± 22, 65.2 ± 31.9, 27.6 ± 8.2, and 15.1 ± 4.1 ng / mL, respectively, after administration of 0.3, 1, 3, 10 and 30 mg / . The results are shown graphically in FIG. 10b.

No damage and testosterone levels in ORX rats

Serum testosterone in the uninjured Beihikle control group was 2.4 ± 1.1 ng / mL. The lower limit of determination for T was 0.08 ng / mL. Values less than 0.08 ng / mL are shown below the quantitation limit (BLOQ). In intact animals, the compound of formula IV decreased T levels dose-dependently and a significant decrease was observed at doses of ≥ 3 mg / kg / day. Testosterone levels in untreated animals treated with compounds of formula IV were 2.6 ± 1.7, 1.6 ± 1.0, 0.7 ± 0.4, BLOQ and BLOQ ng after administration of 0.3, 1, 3, 10 and 30 mg / / mL. Within the ORX animals, the T level was BLOQ in all treated group IV and in the Beckle treated group. The results for uninjured animals are graphically presented in Fig. 10c (and Fig. 2) (indicated as quantitative limits for graphing BLOQ values)

As shown in Figure 9, rapid and potent inhibition of serum testosterone in intact male rats by administering Compound IV at doses of 3 mg / kg, 10 mg / kg and 300 mg / kg after 24h, 74h and 168h Respectively.

Tissue weight (Table 6)

Prostate, seminal vesicle and anal anus. The organ weights (mean ± SD) are presented in Figures 10d, 10e and 10f, respectively. A dose-dependent reduction in prostate, seminal vesicle and anal muscle mass was observed in compound IV treated intact animals. The prostate weights in uninjured animals were 84.0 ± 19.2, 75.2 ± 20.7, 68.2 ± 8.1, 45.1 ± 20.0, and 43.6 ± 8.8 after administration of 0.3, 1, 3, 10 and 30 mg / The prostate weights in ORX animals were 19.0 ± 4.2, 17.4 ± 3.4, 19.6 ± 6.7, 22.9 ± 5.4, and 20.6 ± 2.1 after administration of 0.3, 1, 3, 10 and 30 mg / The mean weight of seminal vesicles in the intact animals was 76.2 ± 7.8, 66.3 ± 27.2, 51.8 ± 28.5, 19.1 ± 7.0, and 17.9 ± 3.3 after administration of 0.3, 1, 3, 10 and 30 mg / The seminal vesicle weight in ORX animals was 12.2 ± 1.3, 16.6 ± 5.4, 16.5 ± 4.8, 13.3 ± 1.9, and 12.9 ± 2.1, respectively, after administration of 0.3, 1, 3, 10 and 30 mg / The weight of intestinal muscle in the intact animals was 86.9 ± 10.0, 82.1 ± 12.1, 65.2 ± 4.4, 57.8 ± 11.2, and 58.1 ± 4.7 after administration of 0.3, 1, 3, 10 and 30 mg / The mean weights of the anal ankles in ORX animals were 4.5 ± 6.6, 49.6 ± 7.0, 53.6 ± 10.0, 51.1 ± 4.9, and 49.2 ± 4.2 after administration of 0.3, 1, 3, 10 and 30 mg / LH inhibition and organ weight data are summarized in Table 6.

In vivo effects of compounds of formula IV on serum hormones and organ weights gonad
condition compound Dose
(mg / kg / day) LH
(ng / mL) FSH
(ng / mL) prostate
(No damage%) Semen
(No damage%) anus
elevator
(No damage%)
Intact Beihik - Average 1.46 ^b 20.9 ^b 100.0 ^b 100.0 ^b 100.0 ^b S.D. 0.642 8.49 28.6 13.4 4.97 ORX Beihik - Average 11.1 ^a 93.5 ^a 13.7 ^a 14.0 ^a 58.8 ^a S.D. 3.87 13.8 2.56 2.93 6.62 Intact Compound IV 0.3 Average 0.863 ^b 17.3 ^b 84.0 ^b 76.2 ^{a, b} 86.9 ^{a, b} S.D. 0.384 6.44 19.2 7.83 10 Intact Compound IV One Average 0.704 ^b 15.7 ^b 75.2 ^b 66.3 ^{a, b} 82.1 ^{a, b} S.D. 0.53 7.26 20.7 27.2 12.1 Intact Compound IV 3 Average 0.395 ^{a, b} 18.4 ^b 68.2 ^{a, b} 51.8 ^{a, b} 65.2 ^a S.D. 0.302 7.72 8.12 28.5 4.35 Intact Compound IV 10 Average 0.226 ^{a, b} 9.25 ^{a, b} 45.1 ^{a, b} 19.1 ^a 57.8 ^a S.D. 0.165 3.97 20 6.98 11.2 Intact Compound IV 30 Average 0.236 ^{a, b} 6.25 ^{a, b} 43.6 ^{a, b} 17.9 ^a 58.1 ^a S.D. 0.176 1.82 8.75 3.33 4.71 ORX Compound IV 0.3 Average 15.4 ^a 116 ^a 19.0 ^{a, b} 12.2 ^a 54.5 ^a S.D. 2.94 17.2 4.19 1.31 6.56 ORX Compound IV One Average 13.5 ^a 114 ^a 17.4 ^a 16.6 ^a 49.6 ^a S.D. 2.18 22.3 3.4 5.36 7.04 ORX Compound IV 3 Average 6.5 65.2 ^a 19.6 ^a 16.5 ^a 53.6 ^a S.D. 5.63 31.9 6.67 4.82 10 ORX Compound IV 10 Average 0.425 ^{a, b} 27.6 ^b 22.9 ^{a, b} 13.3 ^a 51.1 ^a S.D. 0.135 8.16 5.44 1.91 4.88 ORX Compound IV 30 Average 0.368 ^{a, b} 15.1 ^b 20.6 ^{a, b} 12.9 ^a 49.2 ^{a, b} S.D. 0.119 4.11 2.08 2.14 4.21

^a P < Damaged Bechle. ^b P < ORX Bechl

Example 10

Restoration of testosterone levels after inhibition by compound IV in rats and monkeys

The reversibility of chemical castration using compound IV was studied

Materials and Methods:

Thirty-five male Sprague-Dawley rats weighing approximately 200 g were maintained in a 12 hour man / cancer cycle with an unlimited supply of food (2016 Teklad Global 16% Protein Rodent Diet, Harlan, Madison, Wis.) And water. The animal protocol was reviewed and approved by the Animal Experimental Ethics Committee of the University of Tennessee.

Test materials for this study were quantitated and dissolved in PEG 300 (100%) (Acros Organics, NJ) to produce suitable dosage formulations. Animals were randomly assigned to one of the 10 treatment groups (n = 5 animals / group). The treatment groups are listed in Table 7. The animals were housed in groups of 2-3 animals per cage. Group 1 was sacrificed at the start of the study (Day 1) for the measurement of baseline testosterone levels in uninjured animals. Groups 1 to 3, or 30 mg / kg daily for 3 days were administered via oral gavage. Groups 2, 3 and 4 were sacrificed on day 4 to determine maximal testosterone inhibition. Groups 5, 6, and 7 were allowed to recover for 14 days with a drug-free wash period.

group Compound IV P.O . Dose process County 1 - Basal County 2 1 mg / kg for 3 days No recovery County 3 3 mg / kg for 3 days No recovery County 4 30 mg / kg for 3 days No recovery County 5 1 mg / kg for 3 days 14 days recovery County 6 3 mg / kg for 3 days 14 days recovery County 7 30 mg / kg for 3 days 14 days recovery

result:

Serum testosterone levels in uninjured rats were 6.4 ± 3.1 ng / mL at baseline (mean ± SD). Compound IV administered for 3 days at a dose of 3 to 30 mg / kg significantly inhibited serum testosterone levels to 1.47 + -0.26 and 1.62 + -0.49 ng / mL, respectively. No significant inhibition was observed in animals receiving Compound IV at 1 mg / kg for 3 days. Most importantly, serum testosterone levels were 3.3 ± 1.92, 3.00 ± 1.06 and 3.8 ± 1.72, respectively, after 14 days of recovery, in animals receiving each of the compounds at 1, 3 or 30 mg / kg for 3 days , And as shown in FIG. 23, there was no statistically significant difference from the baseline serum testosterone concentration in the uninjured rat.

This study confirms previous findings that Compound IV rapidly suppresses serum testosterone levels in intact male rats. Inhibition of serum testosterone levels was observed in the dose group administered ≥ mg / kg / day for 3 days. No significant decrease in serum testosterone was observed in the 1 mg / kg dose group. However, within the recovery period of 14 days, serum testosterone levels returned to the level of the intact control group. This study shows that pharmacological castration by compound IV in rats is reversible.

The effects of compound IV on inhibition and recovery of testosterone levels in uninjured male monkeys were evaluated along with oral pharmacokinetic studies. Three untreated male cynomolgus monkeys (2-3 weeks old) were administered compound IV by oral gavage at a dose of 30 mg / kg for 7 consecutive days. Blood samples were collected and divided into serum and plasma for the determination of testosterone and compound IV, respectively. The results show that the daily dose of compound IV reduced circulating androgens (mainly testosterone and dihydrotestosterone) levels to less than 47% in all three male monkeys compared to the basal level (2 and 6 day treatment [mean ± SEM], levels of 159 ± 72.5, 997 ± 104, and 852 ± 136 ng / mL, respectively, for baseline values). After 18 days of drug-free recovery, the androgen levels returned to normal and were not significantly different from baseline levels (1757.7 ± 369.5 ng / mL after recovery).

Example 11

Despite LH and testosterone reduction in rats, bone maintenance (Table 8)

The effect of compound of formula IV on bone treatment was studied. The orally administered compound of formula IV completely prevented bone loss associated with LH inhibition in intact male rats. Significant LH reductions were induced by compounds of formula IV at doses of ≥ 10 mg / kg / day in intact animals. At 1 mg / kg / day, the compound of formula IV did not significantly decrease LH, but at this dose there was a significant decrease in prostate, seminal vesicle and anal body mass, indicating that circulating testosterone reduction was associated with these androgen- Quot; However, administration of a compound of formula IV at 1 mg / kg / day maintained canine volume at the level of the uninjured control (measured at the distal femur). When administered at doses of 10 and 30 mg / kg / day, the compound of formula IV significantly increased the bone volume in the distal thighs above that of the uninjured control group. These data indicate that Compound IV increased the marginal bone mineral density (BMD) and bone volume percentage to a dose level that reduced LH levels in intact rat. Data from this study are presented in Table 8.

gonad
condition
compound
Dose
(mg / kg / day)
Bone density
(g / cm ³⁾ Percent bone volume (BV / TV)
(%) prostate
(% Of no damage) Semen
(% Of no damage) anus
elevator
(% Of no damage) FSH
(ng / mL) LH
(ng / mL) Intact Beihik - Average 0.274 ^b 20.2 ^b 100.0 ^b 100.0 ^b 100.0 ^b 9.93 ^b 0.781 ^b S.D. 0.033 3.57 11.1 15.9 11.6 2.94 0.263 ORX Beihik - Average 0.224 ^a 15.4 ^a 14.8 ^a 10.3 ^a 59.4 ^a 117 ^a 22.0 ^a S.D. 0.025 2.6 4.08 0.767 7.26 40.2 5.81 Intact Compound IV One Average 0.273 ^b 20.0 ^b 69.2 ^{a, b} 44.6 ^{a, b} 80.0 ^{a, b} 14.1 ^{a, b} 0.820 ^b S.D. 0.04 4.08 13.5 15.7 6.69 4.07 0.392 Intact Compound IV 10 Average 0.326 ^{a, b} 25.9 ^{a, b} 30.7 ^{a, b} 12.8 ^{a, b} 58.1 ^a 5.48 ^{a, b} 0.060 ^{a, b} S.D. 0.048 4.76 12.4 0.886 9.68 1.97 0.092 Intact Compound IV 30 Average 0.326 ^{a, b} 25.5 ^{a, b} 30.1 ^{a, b} 14.4 ^{a, b} 56.1 ^a 6.32 ^{a, b} 0.078 ^{a, b} S.D. 0.046 4.49 17.4 1.45 4.67 3.4 0.114

^a P < Damaged Bechle. ^b P < ORX Bechl

Example 12

17β-hydroxy-steroid dehydrogenase 5 (17β - HSD5) an impact on the enzyme activity

A member of the HSD family is involved in the conversion of circulating steroids. 17β-HSD5 converts androstendione to testosterone and estrone to estradiol. It is also involved in prostaglandin synthesis. The ability of some selected compounds of the invention to inhibit 17 [beta] -HSD5 activity has been demonstrated.

Way

Human 17? -HSD5 was cloned into the pGEX 4t1 vector and the purified protein was prepared. The purified protein was incubated with representative compounds of the present invention, ¹⁴ C androstenedione and NADPH in a suitable buffer. The synthesized testosterone was extracted with ethyl acetate, air dried, and spotted and flowed on a thin layer chromatography (TLC) plate. TLC was exposed to phosphorimager and the intensity of the testosterone band was quantitated. Indomethacin was used as a positive control (LHRH agonist).

result

Compound IV was tested, which had a partial inhibitory effect on 17? -HSD5 enzyme activity. Indymethacin, a positive control (LHRH agonist), showed strong inhibition of the enzyme as expected, as shown in FIG.

Example 13

Toxicology study

Studies were performed to compare thrombolytic activity of compound IV and diethylstilbestrol (DES, positive control) using in vitro human platelet aggregation assays. Since males are the intended therapeutic group for Compound IV (LH inhibition), blood from healthy male donors was used for the study. Platelet-rich plasma was preincubated with estradiol (E2), DES, compound IV or Bevichlo for 30 seconds and then thrombin (0.3 units) was added to induce platelet aggregation. Studies show that preincubation with DES increases thrombin-induced platelet aggregation by approximately 10-fold. However, Compound IV and estradiol reduced platelet-rich plasma aggregation. These data demonstrate that compound IV reduced in vitro human platelet reactivity compared to DES, suggesting that compound IV has less thromboembolic activity than DES (FIG. 4).

Example 14

Effect of Compound IV on facial flushing

A study was conducted to investigate the effect of compound IV on facial flushing using a morphine-dependent rat model (MD model) developed by Simpkins et al (1983) and demonstrated to have some similarities to postmenopausal flushing. In addition to its similarity to human condition, this animal model has a short test time to become a high throughput screening tool for identifying compounds that can alleviate vasomotor nerves using tail skin temperature (TST). TST probe TA-40 (Data Sciences International, MN) was taped to the tail base and the basal temperature was obtained for 15 minutes. After 15 minutes, animals were treated with naloxone (1 mg / kg, SQ) to reverse the effect of morphine. Tail skin temperature (TST) was measured for 1 hour after naloxone treatment with a sampling period of 5 seconds through the experimental procedure. After data acquisition, the moving average of temperature was recorded and calculated for each animal every 60 seconds and further analyzed. The basal temperature was estimated as the average temperature over 15 minutes prior to naloxone administration. The area under the curve (AUC) was calculated by subtracting all values after naloxone administration using the linear trapezoid method.

Compound IV attenuated flushing within the morphine-discontinuous model (see FIG. 13), and the best results were obtained with 10 mg compound IV. 17 [beta] E2 was used at 5 mg / kg in 100% DMSO.

Example 15

Compound IV &lt; / RTI &gt; DES

Prior to the introduction of LHRH agonists, estrogen predominantly achieved a level of steroidal testosterone by increasing estrogenic activity in the pituitary gland via diethylstilbestrol (DES). DES was equally effective as LHRH agonist in achieving testosterone at the steady level. DES treated patients did not show facial flushing or bone loss, but had gynecomastia at a higher rate than ADT using LHRH agonists. Unfortunately, pure estrogens with potent efficacy such as DES and estradiol are often associated with a high risk of serious cardiovascular and thromboembolic complications, which limits their clinical use. It has been hypothesized that increased risk of complications of venous thromboembolism due to DES is due to cross reactivity with other hormone receptors, but it has not been proven. In vitro studies using human platelets have shown that compound IV has a much lower procoagulant activity than DES. Thus, Compound IV, an ER-alpha selective agonist, can deliver the benefits of LHRH agonists while delivering the prostate cancer benefits of DES without causing osteoporosis or deleterious lipid profiles.

Compound IV was as effective as DES in reducing prostate size in rats (Fig. 11a) and modest increase in prostate size in ORX rats (Fig. 11b).

The difference between DES and Compound IV is shown in Figures 12a-12c, where DES cross-reacts with the glucocorticoid receptor (GR) (Figure 12a) and androgen receptor (AR) (Figure 12b), while Compound IV does not. In addition, DES antagonizes estrogen-related receptor (ERR) transcription activation, whereas compound IV does not. Compound IV did not cross-react with any of the three ERR isoforms (ERR-alpha, ERR-beta and ERR-y) as shown in Figure 12c.

Example 16

Monkey toxicity study - 90 days

I got a nomolgus macaca between colonies of Mauritius origin. A prospective study was designed as a 39 week oral pharmacology and toxicological evaluation of compound IV in male cynomolgus monkey and control (LHRH agonist) as a 13 week interim period. A total of 39 sexually mature male monkeys at 5-8 years of age were randomly assigned to Group 5 before initiation of treatment. The groups included 1) a Becky's control, 2) 1 mg / kg Compound IV, 3) 10 mg / kg Compound IV, 4) 100 mg / kg Compound IV and 5) a positive control (LHRH agonist). The drug was administered orally to cervical cancer patients by cage-side administration once daily for 39 weeks using Becky's control compound (Tween 80 / PRANG ^TM ) for Groups 1 and 5, or compounds 2, 3 and 4, . The dose levels of Compound IV were 1, 10 and 100 mg / kg / day for Groups 2, 3 and 4, respectively. Oral administration was calculated based on the most recent useful body weight for each animal and delivered at a dose of 10 mL / kg (FIG. 14). Group 5 animals were also subcutaneously injected (0.02 mL constant dose) once a day in a positive control (LHRH agonist) for a 39-week study period. General appearance and clinical signs were observed and recorded daily. Periodic evaluation and other studies Screening was performed as directed in the study protocol. The screening parameters include, but are not limited to, testosterone, prostate specific antigen (PSA), and prostate volume and weight.

Testosterone and total PSA levels were quantified (using standard procedures) in serum samples using enzyme immunoassay (EIA) and chemiluminescence immunoassay (LIA, ALPCO Diagnostics, Salem NH). Blood samples for testosterone evaluation were collected from all animals (fasted state) at baseline (i. E., Prior to initiation of treatment) and at 1, 3, 7, 14, 28, 64 and 90 days. Blood samples for PSA measurements were collected from all animals (fasted state) at baseline and for 6 weeks. For the sake of discussion, the results of the samples with less than the quantitation limit (BLQ) concentration for testosterone and PSA analysis were calculated as one half of the lower limit of the assay (LLOQ) and considered as the "final concentration estimate". The data in Tables 9 to 16 are presented as "quantifiable concentration only" (ie, excluding the BLQ value) and "final concentration estimate" (ie, samples with BLQ results included as 1/2 LLOQ of the analysis). Prostate volume was measured in live animals under anesthesia at baseline and at 6 weeks using the transrectal ultrasound (RTUS) procedure. The width and height of the prostate gland were recorded. Prostate volume was calculated as width x width x height x pi / 6 and normalized for body weight. The non-fat tissue and the exogenous tissue were cut and the wet weight of the prostate was recorded at the time of autopsy.

Results and discussion:

Serum testosterone levels are presented in Figure 15 and Tables 9-12. At baseline, testosterone levels for all monkeys studied were within normal ranges for sexually mature adult male cynomolgus monkeys. However, testosterone levels were significantly reduced in monkeys receiving compound IV at 100 mg / kg / day and monkeys treated with a positive control (LHRH agonist). Testosterone levels in the positive control (LHRH agonist) were significantly higher (p <0.01) at the initial 47.4% and at the 1st and 4th days, respectively, by 47.4% and 547% (p <0.01) Phase changes of 3.6%, 67%, 73%, 83% and 85% reduction on days 28, 64 and 90 (see FIG. 15 and Tables 9 to 12). A similar immediate effect was not observed at the highest dose level (i.e., 100 mg / kg / day) for compound IV treated animals, and a dose of 100 mg / kg / 60%, 51%, 42%, 79% and 92%, respectively, on days 7, 14, 28 and 64 (see FIG. 15 and Tables 9 and 10). After treatment with 100 mg / kg / day of compound IV for 90 days, testosterone levels were reduced to sub-limit quantities of analysis within 6-10 of the 4 monkeys in group 4 (see Table 11). Mean serum testosterone levels in group 4 monkeys were reduced by 96% compared to respective basal levels ("final concentration estimates", ie, testosterone levels for 6/10 monkeys with BLQ values were calculated as 50% of the LLOQ concentration, See Table 10). It is important to note that 100 mg / kg / day of Compound IV by 90 days significantly reduced serum testosterone to a significantly lower level than the positive control (LHRH agonist) (p = 0.013).

Compound IV Mean serum testosterone level (ng / mL) in uninjured male monkey after daily oral administration; ^@ Final concentration estimate. Work
Beihik
Control group Compound IV
1 mg / kg Compound IV
10 mg / kg Compound IV
100 mg / kg Positive control group
(LHRH agonist) Average SEM N Average SEM N Average SEM N Average SEM N Average SEM N 0
Basal 6.1 1.2 10 7.3 1.0 6 4.9 0.6 6 4.4 0.6 10 4.9 0.9 7 One 8.0 1.7 10 11 1.6 6 7.6 1.1 6 8.0 2.2 10 7.2 0.8 7 3 8.2 2.3 10 7.4 1.2 6 5.1 1.1 6 1.8 ^* 0.5 10 32 ^{# $} 3.8 7 7 5.9 1.2 10 6.7 0.8 6 7.7 1.9 6 2.2 ^* 0.7 9 4.7 2.6 7 14 3.4 0.5 10 3.8 0.4 6 7.1 1.6 6 2.6 0.9 9 1.6 ^# 0.2 7 28 3.8 0.6 10 4.7 0.9 6 9.4 2.1 6 0.9 ^* 0.2 10 1.3 ^# 0.2 7 64 5.1 1.1 10 4.3 0.6 6 5.4 1.5 6 0.3 ^* 0.1 9 0.8 ^{# $} 0.2 7 90 3.6 0.6 9 4.2 0.6 4 4.6 1.0 5 0.2 ^* 0.0 10 0.8 ^{# $} 0.2 7

Testosterone assay LLOQ = 0.246 ng / mL; The BLOQ value is 0.123 ng / mL, which is half of the LLOQ.

*: Statistically significant (p < 0.05) Compound IV 100 mg / kg vs. Beihikle control

#: Statistically significant (p <0.05) positive control (LHRH agonist) vs. Beihikle control

$: Statistically significant (p <0.05) positive control (LHRH agonist) vs. Compound IV 100 mg / kg

Percentage change in mean serum testosterone level compared to baseline (%); ^@ Final concentration estimate. Work Beihik
Control group Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg Positive control group One 31 44 54 82 47 3 35 1.8 3.5 -60 547 7 -3.2 -8.1 57 -51 -3.6 14 -44 -48 45 -42 -67 28 -38 -35 92 -79 -73 64 -16 -41 11 -92 -83 90 -42 -42 -5.5 -96 -85

Testosterone assay LLOQ = 0.246 ng / mL; The BLOQ value is 0.123 ng / mL, which is half of the LLOQ.

Mean serum testosterone levels (ng / mL) in uninjured male monkeys after oral administration of Compound IV daily; ^λ Quantifiable concentration only Work Beihikle control Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg Positive control group
(LHRH agonist) Average SEM N Average SEM N Average SEM N Average SEM N Average SEM N 0
Basal 6.1 1.2 10 7.3 1.0 6 4.9 0.6 6 4.4 0.6 10 4.9 0.9 7 One 8.0 1.7 10 11 1.6 6 7.6 1.1 6 8.0 2.2 10 7.2 0.8 7 3 8.2 2.3 10 7.4 1.2 6 5.1 1.1 6 1.8 0.5 10 32 3.8 7 7 5.9 1.2 10 6.7 0.8 6 7.7 1.9 6 2.2 0.7 9 4.7 2.6 7 14 3.4 0.5 10 3.8 0.4 6 7.1 1.6 6 2.6 0.9 9 1.6 0.2 7 28 3.8 0.6 10 4.7 0.9 6 9.4 2.1 6 0.9 0.2 10 1.3 0.2 7 64 5.1 1.1 10 4.3 0.6 6 5.4 1.5 6 0.3 0.1 9 0.8 0.2 7 90 3.6 0.6 9 4.2 0.6 4 4.6 1.0 5 0.2 0.1 4 0.8 0.2 7

Testosterone assay LLOQ = 0.246 ng / mL; ^λ BLQ value is excluded.

Percent change in mean testosterone level compared to baseline (%); ^λ Quantifiable concentration only Work Beihik
Control group Compound IV
1 mg / kg Compound IV
10 mg / kg Compound IV
100 mg / kg positivity
Control group One 31 44 54 82 47 3 35 1.8 3.5 -60 547 7 -3.2 -8.1 57 -51 -3.6 14 -44 -48 45 -42 -67 28 -38 -35 92 -79 -73 64 -16 -41 11 -92 -83 90 -42 -42 -5.5 -95 -85

Testosterone assay LLOQ = 0.246 ng / mL; ^λ BLQ value is excluded.

Serum PSA levels were also significantly inhibited by Compound IV within 4 weeks of treatment initiation. PSA levels in the positive control (LHRH agonist) were reduced by 60%, while 69% and 87% (mean) PSA reduction was noted for monkeys given compound IV at 10 mg / kg and 100 mg / kg for 4 weeks (Fig. 16 and Table 13-16).

Mean serum PSA level (ng / mL) in uninjured male monkey after daily oral administration of compound IV; ^@ Final concentration estimate. Beihikle control Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg The positive control (LHRH agonist) Average SEM N Average SEM N Average SEM N Average SEM N Average SEM N Before administration 1.1 0.2 10 1.0 0.2 6 0.8 0.1 6 1.0 0.1 10 1.0 0.1 7 4 weeks 1.0 0.2 10 0.9 0.2 6 0.3 * 0.1 6 0.1 ^& 0.1 10 0.4 ^{# $} 0.1 7

PSA assay LLOQ = 0.0575 ng / mL; ^{The @} BLQ value is calculated as 0.02875 ng / mL, half the LLOQ.

*: Statistically significant (p < 0.05) Compound IV 10 mg / kg vs. Beihikle control

& Statistically significant (p < 0.05) Compound IV 100 mg / kg vs. Beihikle control

#: Statistically significant (p <0.05) positive control (LHRH agonist) vs. Beihikle control

$: Statistically significant (p <0.05) positive control (LHRH agonist) vs. Compound IV 100 mg / kg

Percentage change in average PSA level compared to baseline (%); ^@ Final concentration estimate. Control group Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg The positive control (LHRH agonist) 4-week -7.1 -11 -69 -87 -60

PSA assay LLOQ = 0.0575 ng / mL; ^{The @} BLQ value is calculated as 0.02875 ng / mL, half the LLOQ.

Mean serum PSA level (ng / mL) in uninjured male monkey after daily oral administration of compound IV; ^λ Quantifiable concentration only. Beihikle control Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg The positive control (LHRH agonist) Average SEM N Average SEM N Average SEM N Average SEM N Average SEM N Before processing 1.2 0.2 9 1.0 0.2 6 0.8 0.1 6 1.0 0.1 10 1.0 0.1 7 4-week 1.1 0.1 9 0.9 0.2 6 0.3 0.1 5 0.3 0.1 4 0.4 0.1 7

PSA assay LLOQ = 0.0575 ng / mL; ^λ BLQ values are excluded from the table above.

Percentage change in average PSA level compared to baseline (%); ^λ Quantifiable concentration only. Control group Compound IV
1 mg / kg Compound IV
10 mg / kg Compound IV
100 mg / kg Positive control group 4 weeks -7.1 -11 -64 -72 -60

PSA assay LLOQ = 0.0575 ng / mL; ^λ BLQ values are excluded from the table above.

Prostate volume was measured periodically by TRUS through studies. The results obtained after six weeks of treatment demonstrated a potent effect of compound IV and a positive control (LHRH agonist) on the monkey prostate. Compound IV reduced the prostate volume by 25% and 45% at the 10 mg / kg and 100 mg / kg dose levels, respectively, while the prostate volume was reduced by 28% within the positive control (LHRH agonist) 17 and 18).

The average prostate volume (ratio) in male monkey after daily oral administration of compound IV Beihikle control Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg The positive control (LHRH agonist) Average SEM N Average SEM N Average SEM N Average SEM N Average SEM N 6 weeks 438 78 10 468 78 6 327 33 6 242 28 10 315 47 7

Percentage change in mean prostate volume compared to baseline (%) Control group Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg positivity
Control group 6 weeks 0 6.8 -25 -45 -28

A decrease in prostate volume IV-related prostate volume was determined by autopsy weight assessment at autopsy. After 13 week treatment, compound IV significantly reduced the mean prostate weight by 24% and 21% in the animals receiving 10 and 100 mg / kg / day, respectively (Fig. 18b and Tables 19 and 20).

The average prostate weight (g) at the time of autopsy in male monkey after daily oral administration of compound IV Beihikle control Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg Average SEM N Average SEM N Average SEM N Average SEM N 13 weeks 1.8 0.2 3 1.8 0.4 3 1.3 0.1 3 1.4 0.1 3

Percent change in average prostate weight compared to baseline (%) Control group Compound IV 1 mg / kg Compound IV 10 mg / kg Compound IV 100 mg / kg 13 weeks 0 1.7 -24 -21

No apparent effect on platelet aggregation, prothrombin time (PT) or activated partial thromboplastin time was observed.

Example 17

Study of compound IV in humans

Studies were conducted to determine the effect of compound IV on human males. Twelve subjects per group were studied at doses of 100, 300, 600 and 1000 mg of compound IV. Table 21 presents the mean changes in LH, serum PSA, free testosterone, and total testosterone levels in men upon administration of Compound IV at doses of 100, 300, 600 and 1000 mg. A dose-dependent mean total testosterone level (nmol / L) was measured in males during the period of 1-11 (Figure 19). Total testosterone levels were reduced by 51.9% and 47.9% at doses of 600 mg and 1000 mg, respectively.

Dose-dependent mean LH levels (IU / L) in humans were measured over a period of 1-10 days (Figure 20). LH levels increased by 20.7%, 46.9%, 27.6% and 29.2% at doses of 100 mg, 300 mg, 600 mg and 1000 mg, respectively.

Dose-dependent mean free testosterone levels (pg / mL) in humans were measured over a period of 1-10 days (Figure 21). Free testosterone levels were reduced by 17.0%, 18.5%, 72.7% and 53.2% at doses of 100 mg, 300 mg, 600 mg and 1000 mg, respectively.

A dose-dependent mean PSA level ([mu] g / L) was measured in humans for a period of 1-10 days (Figure 22). PSA levels were reduced by 9.2%, 24.4%, 27.5% and 29.9% at doses of 100 mg, 300 mg, 600 mg and 1000 mg, respectively. No changes were noted at doses of 10 and 30 mg.

Mean change from baseline 100 mg 300 mg 600 mg 1000 mg Serum PSA -9.2% -24.4% -27.5% -29.9% LH 20.7% 46.9% 27.6% 29.2% freedom
Testosterone -17.0% -18.5% -72.7% -53.2% Total testosterone 3.9% 7.3% -51.9% -47.9%

Example 18

Bioavailability of compound IV

Compound IV was rapidly absorbed after oral administration to rats, dogs and monkeys. The oral bioavailability of Compound IV in rats was between 6% and 25%, depending on the formulation being administered. Preparations using polyethylene glycol 300 (PEG 300) generally produced higher exposures than microemulsions prepared in diluted Tween 80 in deionized water. Within the dog, visual inspection of the plasma concentration-time profile suggests that compound IV undergoes long-term recycling, as evidenced by the second peak at the end. It is important within the dogs that exposure in the male 30 mg / kg PEG 300 oral administration group would exceed the exposure required to produce the maximal effect on prostate reduction in the LH-inhibitory rat model. Within monkeys, preliminary pharmacokinetic studies suggest that oral bioavailability in these species is similar to or greater than that in dogs, as evidenced by plasma levels of compound IV and serum testosterone inhibition over a 7 day period. Overall, this data suggests that sufficient oral exposures may be achieved to produce the desired pharmacological effect (based on AUC data) within the two non-rodent species. Furthermore, endocrine data in rats and monkeys suggest that the pharmacological effects of compound IV are reversible (i. E. Serum concentration of testosterone returns to basal or normal levels upon discontinuation of compound IV treatment).

Example 19

Pharmacokinetics of Compound IV

Preliminary data from in vitro (mouse, rat, dog, monkey and human) and in vivo (rat) metabolism studies suggest that the conjugation of compound IV, its hydroxylated metabolite (s) and its N-dealkylated metabolite And the overall placement of compound IV in humans. Interspecific comparison results show that, although only qualitatively, the overall metabolite profile of the non-clinical species adequately reflects the profile generated in human liver microsomes. Based on these results, rats and dogs are suitable rodent and non-rodent species for pharmacological and toxicological evaluation. In vitro studies show that compound IV does not induce CYP450 isoforms (CYP1A2, CYP2B6, or CYP3A4) and does not inhibit CYP1A2, CYP2C19, CYP2D6, or CYP3A4 / 5 at a concentration of <30 μM. CYP2C9 is inhibited by compound IV, but only at high concentrations (Ki = 8 μM) and the possibility of pharmacokinetic drug-drug interactions is considered to be sparse.

Example 20

Biological activity of compound IV

Compound IV hardly exerts or exerts an in vitro inhibitory effect (IC ₅₀ ? 30 μM) on the hERG channel. The compounds reduced APD50 and APD90 in a dose dependent manner at concentrations of 10 and 100 [mu] M in in vitro isolated Purkinje fibers. However, Compound IV did not affect hemodynamic or cardiac function (blood pressure, heart rate, electrocardiographic morphology or QT interval) in remotely-measured dogs at any dose (up to 300 mg / kg). No neuropharmacological or lung effects were observed. No significant effect on renal function was noted with a single oral administration of Compound IV at 30 mg / kg or less. Only the urine volume and urinary excretion of potassium and chloride were observed at the highest dose tested (100 mg / kg). Oral administration of Compound IV at doses of 30 to 300 mg / kg in the rats resulted in a significant increase in peritoneal motility, and oral administration of 30 mg / kg of Compound IV in rats resulted in a significant increase in gastrointestinal motility and gastric acidity (Not likely to be due to an effect on smooth muscle).

Compound IV was not a mutagenic agent and did not induce structural or aberrant chromosomal aberrations at concentrations below 200 [mu] M in vitro in human peripheral blood lymphocytes. Rats and dogs survived well after single and repeated oral administration (up to 28 days) of Compound IV. No pathological changes were observed in the kidney, liver, heart, and other non-target related organs. There were no significant physical signals, weight effects, clinical pathologic changes, ophthalmological, electrocardiographic or histopathological changes associated with oral administration of male or female dogs of compound IV for periods of up to 28 days.

While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will occur to those skilled in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims (14)

(PSA) level in a male subject, comprising administering a therapeutically effective amount of a compound of formula I, an isomer thereof, a pharmaceutically acceptable salt, a pharmaceutical product, a hydrate or a combination thereof, &Lt; RTI ID = 0.0 &gt;

In this formula,
Y is C (O) or CH ₂ ;
R ₁ and R ₂ are each independently selected from the group consisting of hydrogen, halogen, hydroxyl, alkoxy, cyano, nitro, CF ₃ , N (R) ₂ , sulfonamide, SO ₂ R, alkyl, haloalkyl, aryl, O- NR ₅ R ₆ Or O-Alk-heterocycle, wherein said heterocycle is optionally aromatic and is a 3-7 membered substituted or unsubstituted heterocycle ring;
R ₃ and R ₄ is each independently selected from the group consisting of hydrogen, halogen, hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF ₃ , NHCOR, N (R) ₂ , sulfonamide, SO ₂ R, alkyl, haloalkyl, Lt; / RTI &gt;
R is alkyl, trihalomethyl alkyl, hydrogen, haloalkyl, dihalo, CH ₂ F, CHF ₂ , CF ₃ , CF ₂ CF ₃ , aryl, phenyl, halogen, alkenyl, CN, NO ₂ , or OH;
R ₅ and R ₆ are independently hydrogen, phenyl, alkyl of 1-6 carbon atoms, 3-7 membered cycloalkyl, 3-7 membered heterocycle, 5-7 membered aryl; R &lt; ₅ &gt; and R &lt; ₆ &gt; together with the nitrogen atom form a 3-7 membered ring;
j and k are independently 1 to 4; And
Alk is linear alkyl of 1-7 carbons, branched alkyl of 1-7 carbons, or cyclic alkyl of 3-8 carbons. The method according to claim 1,
Wherein the compound of formula I is selected from:

or

The method according to claim 1,
Wherein the subject has a progressive prostate cancer. The method according to claim 1,
Wherein the subject has undergone androgen blockade therapy (ADT). The method according to claim 1,
Wherein the administration of said compound does not cause side effects associated with androgen blockade therapy (ADT). 6. The method of claim 5,
Wherein said side effect is selected from the group consisting of facial flushing, gynecomastia, increased body fat, bone loss, decreased bone density, and increased risk of fracture. The method according to claim 1,
Wherein said compound, its isomer, pharmaceutically acceptable salt, pharmaceutical product, hydrate or combination thereof is administered at a dose of 125 mg, 250 mg or 500 mg per day. (SHBG) in a subject comprising a therapeutically effective amount of a compound of formula I, an isomer thereof, a pharmaceutically acceptable salt, a pharmaceutical product, a polymorph, a hydrate or a combination thereof, Method to increase:

In this formula,
Y is C (O) or CH ₂ ;
R ₁ and R ₂ are each independently selected from the group consisting of hydrogen, halogen, hydroxyl, alkoxy, cyano, nitro, CF ₃ , N (R) ₂ , sulfonamide, SO ₂ R, alkyl, haloalkyl, aryl, O- NR ₅ R ₆ Or O-Alk-heterocycle, wherein said heterocycle is optionally aromatic and is a 3-7 membered substituted or unsubstituted heterocycle ring;
R ₃ and R ₄ is each independently selected from the group consisting of hydrogen, halogen, hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF ₃ , NHCOR, N (R) ₂ , sulfonamide, SO ₂ R, alkyl, haloalkyl, Lt; / RTI &gt;
R is alkyl, trihalomethyl alkyl, hydrogen, haloalkyl, dihalo, CH ₂ F, CHF ₂ , CF ₃ , CF ₂ CF ₃ , aryl, phenyl, halogen, alkenyl, CN, NO ₂ , or OH;
R ₅ and R ₆ are independently hydrogen, phenyl, alkyl of 1-6 carbon atoms, 3-7 membered cycloalkyl, 3-7 membered heterocycle, 5-7 membered aryl; R &lt; ₅ &gt; and R &lt; ₆ &gt; together with the nitrogen atom form a 3-7 membered ring;
j and k are independently 1 to 4; And
Alk is linear alkyl of 1-7 carbons, branched alkyl of 1-7 carbons, or cyclic alkyl of 3-8 carbons. 9. The method of claim 8,
Wherein the compound of formula I is selected from:

or

9. The method of claim 8,
Wherein the subject has a progressive prostate cancer. 9. The method of claim 8,
Wherein the subject has undergone androgen blockade therapy (ADT). 9. The method of claim 8,
Wherein the administration of said compound does not cause side effects associated with androgen blockade therapy (ADT). 13. The method of claim 12,
Wherein said side effect is selected from the group consisting of facial flushing, gynecomastia, increased body fat, bone loss, decreased bone density, and increased risk of fracture. 9. The method of claim 8,
Wherein said compound, its isomer, pharmaceutically acceptable salt, pharmaceutical product, hydrate or combination thereof is administered at a dose of 125 mg, 250 mg or 500 mg per day.

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