MX340753B

Estrogen receptor ligands and methods of use thereof.

Abstract

The present invention relates to methods for reducing testosterone levels by reduction of luteinizing hormone (LH) or independent of LH levels in a male subject and methods of treating, suppressing, reducing the incidence, reducing the severity, or inhibiting advanced prostate cancer and palliative treatment of advanced prostate cancer.

MX340753B, drawing sheet 1 — Sheet 1 of 161

Term

No projected expiry on record.

3 claims: 2 independent, 1 dependent

1
CLAIMS REIVINDICACIONES 1. Uso de un compuesto agonista de ER-alfa de fórmula I, o su isómero, sal farmacéuticamente aceptable, producto farmacéutico, polimorfo, hidrato o cualquier combinación de los mismos, en la elaboración de un medicamento para el tratamiento de cáncer avanzado de próstata en un sujeto masculino, en dond dicho compuesto agonista de ER-alfa de fórmula I se representa por la estructura:one. Use of an ER-alpha agonist compound of formula I, or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate, or any combination thereof, in the manufacture of a medicament for the treatment of advanced prostate cancer in a male subject, where said ER-alpha agonist compound of formula I is represented by the structure: en donde where Y es C(O);Y is C (O);R! and R2 they are independently hydrogen, halogen, hydroxyl, alkoxy, cyano, nitro, CF3, N (R)2, alkyl, haloalkyl, or aryl;R! y R2 son independientemente hidrógeno, halógeno, hidroxilo, alcoxi, ciano, nitro, CF3, N(R)2, alquilo, haloalquilo, o arilo;R3 and R4 they are independently hydrogen, halogen, hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF3, NHCOR, N (R)2, alkyl, haloalkyl, aryl, or protected hydroxyl;R3 y R4 son independientemente hidrógeno, halógeno, hidroxialquilo, hidroxilo, alcoxi, ciano, nitro, CF3, NHCOR, N(R)2, alquilo, haloalquilo, arilo, o hidroxilo protegido;R es alquilo, hidrógeno, haloalquilo, d i ha loa I q u i lo, trihaloalquilo, CH2F, CHF¡>, CF3, CF2CF3, arilo, fenilo, halógeno, alquenilo, CN, NO2, u OH;y j y k son ind p ndlent mente 1-4;y R is alkyl, hydrogen, haloalkyl, di ha loa I qui lo, trihaloalkyl, CH2F, CHF¡>, CF3, CF2CF3, aryl, phenyl, halogen, alkenyl, CN, NO2, or OH;and j and k are ind p ndlent mind 1-4;and 137 137 IMPI ^ IMPI^ INSTITUTE Μ EXICANO _t, INSTITUTO Μ EXICANO _t, DELA PROPIEDAD OF THE PROPERTY INDUSTRIAL ^ ΝβΤ * · *. INDUSTRIAL ^ΝβΤ*·*. en donde dicho cáncer avanzado de nr<s»tata pr nánnar da próstata metastásico, y en donde dicho tratamiento de cánc r avanzado de próstata es mediante la reducción del nivel de testosterona total en suero o los niveles de testosterona libr en suero en dicho sujeto. wherein said advanced cancer of nr <s »tata prnánnar gives metastatic prostate, and where said treatment of advanced prostate cancer is by reducing the serum total testosterone level or serum libr testosterone levels in said subject .
3
The use according to claims 1 or 2, wherein the serum total testosterone level is decreased below about 100 ng / dL, 50 ng / dL or 25 ng / dL. 3. El uso de acuerdo con las reivindicaciones 1 o 2, en donde el nivel de testosterona total en el suero es disminuida debajo de aproximadamente 100 ng/dL, 50 ng/dL o 25 ng/dL. io io 139 139

Independent claims2

1,017 paragraphs in 161 sections, as filed

(54) Title: STROGEN RECEIVER LINKS AND METHODS FOR USING THEM. (54) Title: ESTROGEN RECEPTOR LIGANDS AND METHODS OF USE THEREOF.

(57) Summary

The present invention relates to methods of reducing testosterone levels through reduction of luteinizing hormone (LH) or independent of LH levels in a male subject and methods of treating, suppressing, reducing incidence, reducing severity , or inhibit advanced prostate cancer and palliative treatment of advanced prostate cancer.

(57) Abstract

The present invention relates to methods for reducing fesfosferone levels by reducfion of lufeinizing hormone (LH) or independenf of LH levels in a male subjecf and methods of freafing, suppressing, reducing the incidence, reducing the severify, or inhibiting advanced prostafe cancer and palliafive treatment of advanced prostate cancer.

Institute

Mexican Property

Industrial

PATENT TITLE NO. 340753 _SE_ aefttTAMA Bl KONOMY

Owner (s): θΤΧ, INC.

Address: 175 Toyota Plaza, 7th Floor, Memphis, Tennessee, 38103, USA

Name: STROGEN RECEIVER LEAGUES AND METHODS FOR USING THEM.

Classification: lnt.CI.8: A01N43 / 40; A61K31 / 445; A61K31 / 55

Inventor (s): JAMES T. DALTON; MITCHELL S. STEINER

Number:

MX / a / 2011/008879

Country:

REQUEST

International filing date:

February 2010

PRIORITY

Date: Number:

US

US February 2009 April 14, 2009 November 16, 2009

61/154,707

61/168,983

61/261,669

Validity: Twenty years

Expiration Date: February 23, 2030

The reference patent is granted based on articles 1, 2 fraction V, 6 fraction III, and 59 of the Industrial Property Law

Pursuant to article 23 of the Industrial Property Law, this patent has a non-extendable term of twenty years, counted from the date of filing of the international application and will be subject to the payment “fe la tatito” to keep the rights.

Whoever subscribes to the preafente title Jo does based on the provisions of articles β ° fractions lll and 7 ° bis 2 of the Industrial Property Law (Official Letter of the Federation (DOF) 27/06 / 19S1, re-issued on 02 / 08/1994 10/25/1996, 12/26/1967, 05/17/1999, 01/26/2004, 06/16/2005, 01/25/2006, 06/05/2009, 01/06/2010 06/18 / 1C10 78 / 06,714,? »U * 201? , 04/09/2012); 1st articles. 3rd fraction V subsection a), 4th and 12th fraqjiones I jf lll of the Regulations of the Institute of Industrial Property (DOF 14/12/1960, amended on 07/01/2002, 07/15/2007% 28 / 07/2604 and 7/09/2007); Articles 1, 3, 4, 5, subsection a), 16 sections I and III and 30 of the Organic Document of the Mexican Institute of Industrial Property (OOF) 12/27/1999, amended on 10/10/2002, 07/29/2004, 04.08 / 2004 and 09/13/2007), 1st, 3rd and 5th paragraph a) of the Agreement that delegates powers to the Deputy Directors General, Coordinator, Division Directors, Head of the Regional Offices, Divisional Deputy Directors, Departmental Coordinators and other subordinates of the Mexican Institute of Industrial Property (DOF 12/15/1999, amended on 02/04/2000, 07/29/07 / 2004, 08/04/2004 and 09/13/2007).

Issue Date: July 25, 2013

THE DIVISIONAL DIRECTOR OF PATENTS

NAHANNY CANAL REYES / Vena! No $ 50 Floor 1. I heard. Puebíc Santa María Tepepan.

xochirntico. c Η ΐΰϋζΰ Mexico City

Tel. (55 '53 34 07 00 www impi qob.m *

MX / 2016/57971

2.ΠΙ)

ESTROGEN RECEPTOR LEAGUARDS AND METHODS OF

USE OF THE SAME

FIELD OF THE INVENTION

The present invention relates to methods of reducing testosterone levels by reducing luteinizing hormone (LH) or LH-independent hormone levels in a male subject, and methods of treating, suppressing, and reducing incidence, reducing severity, or inhibiting advanced prostate cancer and palliative treatment for advanced prostate cancer.

BACKGROUND OF THE INVENTION

Estrogens refers to a group of endogenous and synthetic hormones that are important to and used for the maintenance of tissue and bones. Estrogens are endocrine regulators in the cellular processes involved in the development and maintenance of the reproductive system. AND! The role of estrogens in reproductive biology, the prevention of postmenopausal hot flashes, and the prevention of postmenopausal osteoporosis is well established. Estradiol is the main endogenous human estrogen, and is found in both women and men.

The biological actions of estrogens and antiestrogens are manifested through two different intracellular receptors.

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alpha estrogen receptor (ERa) and beta estrogen receptor (ER3).

Endogenous estrogens are typically potent 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 commonly act differently on different tissues. For example, a SERM may be an ERa antagonist in the breast, but it may be a partial ERa agonist in the uterus, bones, and cardiovascular system. Compounds that act as estrogen receptor ligands, therefore, are useful in treating a variety of conditions and disorders.

Prostate cancer is one of the most frequently diagnosed non-skin cancers among men in the United States and is the second most common cause of cancer deaths with over 180,000 new cases and nearly 29,000 deaths this year. Patients with advanced prostate cancer undergo androgen deprivation therapy (ADT), typically either using luteinizing hormone-releasing hormone (LHRH) agonists or bilateral orchidectomy. Androgen deprivation therapy not only lowers testosterone, but also lower estrogen levels since estrogen is derived from aromatization of testosterone, these levels are decreased by ADT. Estrogen deficiency induced by androgen deprivation therapy causes significant side effects, which

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MEXICAN INSTITUTE

OE THE PROPERTY

INDUSTRIAL nr »include hot flashes, gynecomastia and mastalgia, bone loss, reductions in bone quality and strength, osteoporosis and life-threatening fractures, adverse lipid changes and superior cardiovascular disease and myocardial infarction, and depression and other changes in the mood. Many of the estrogen deficiency side effects of ADT are believed to be mediated by ERa.

Leuprolide Acetate (Lupron®) is a synthetic non-peptidic analog of naturally occurring gonadotropin-releasing hormone (GnRH or LH-RH). Leuprolide Acetate is an LH-RH super antagonist that eventually suppresses LH secretion by the pituitary. Leuprolide acetate acts as a potent inhibitor of gonadotropin secretion, resulting in suppression of testicular and ovarian steroidogenesis. In humans, administration of leuprolide acetate results in an initial increase in circulating levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH), leading to a transient increase in the levels of gonadal steroids (testosterone and dihydrotestosterone in men, and estrone and estradiol in premenopausal women). However, continuous administration of leuprolide acetate results in reduced levels of LH and FSH. In men, testosterone is reduced to castration levels (below 50 ng / dL). In premenopausal women, estrogens are reduced to postmenopausal levels. Testosterone is a stimulus

known for cancer cells of the prostate. Suppressing testosterone secretion or inhibiting testosterone actions in this way is a necessary component of prostate cancer therapy. Leuprolide acetate can be used to suppress LH, which is the reduction and decrease of serum testosterone to castrating levels to treat prostate cancer.

Prior to the introduction of LHRH agonists, castration testosterone levels were achieved by increasing estrogen activity in the pituitary through estrogens, primarily diethylIbestosterI (DES). DES was equally effective as LHRH agonists in suppressing testosterone at castration levels. Patients treated with DES had no hot flashes or bone loss, but had gynecomastia at higher rates than ADT with LHRH agonists. Unfortunately, highly potent estrogens, such as DES and estradiol, are generally associated with a high risk of severe cardiovascular and thromboembolic complications that have limited their clinical use.

The compounds of this invention suppress testosterone levels to castration levels, which can be used to treat prostate cancer, while avoiding the high risk of thrombotic events, and without causing bone loss, hot flashes and / or gynecomastia.

MEXICAN INSTITUTE

OF IA PROPERTY

INDUSTRIAL

BRIEF DESCRIPTION OF THE INVENTION

In one embodiment, this invention provides a method of reducing serum total testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a compound of formula I-XII as described below, wherein the reduction of Total serum testosterone occurs by a reduction in serum luteinizing hormone levels.

In one embodiment, this invention provides a method of reducing serum total testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a compound of formula I-XII as described below, wherein the reduction of Total serum testosterone is independent of a reduction in serum luteinizing hormone levels.

DC INDUSTRIAL rRlWF.DAD is described below, where said administration of said compounds of formula l-XII avoids or tries to avoid side effects associated with androgen deprivation therapy (ADT), where said subject has prostate cancer .

In one embodiment, this invention provides a method for androgen deprivation therapy in a subject, which comprises administering a therapeutically effective amount of a compound of formula l-XIl as described below. In another embodiment, said subject has prostate cancer.

In one embodiment, this invention provides a method of treating, suppressing, reducing the incidence, reducing the severity, or inhibiting advanced prostate cancer, which comprises administering a therapeutically effective amount of a compound of formula IXII as described herein below.

In one embodiment, this invention provides a method of palliative treatment for advanced prostate cancer, comprising a therapeutically effective amount of a compound of formula I-XII as described herein below.

BRIEF DESCRIPTION OF THE DRAWINGS

The subject matter considered as the invention is particularly pointed out and indiscriminately claimed in the concluding portion of the specification. The invention, however, both as an organization and as a method of operation, together with objects,

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MEXICAN INSTITUTE OC THE INDUSTRIAL PROPERTY

characteristics, and advantages thereof, can be better understood by referring to the following detailed description read with the accompanying drawings in which:

Figure 1 shows serum testosterone levels (solid line and androgen totals (dotted line) in intact male monkeys after daily oral administration of 30 mg / kg of Compound IV (first dose on Day 0). (See Examples 8).

Figure 2 shows testosterone levels in rats intact with Compound IV (0.3, 1, 10, 30 mg / kg). 1 denotes P <0.05 against intact vehicle controls. The BLOQ values are graphically represented at the 0.08 ng / ml quantification limit. (See Example 9).

Figure 3 shows the inhibitory effect of Compound IV on the activity of the 17p-HSD5 enzyme. (See Example 12).

Figure 4 shows in vivo aggregation of human platelets in the presence of DES, 1 7β-strdio I (E2), and Compound, IV. Platelet rich plasma (PRP) was incubated with vehicle, E2, DES, or Compound IV for 30 seconds before inducing aggregation with 0.3 thrombin units. Aggregation was verified for 5 minutes and expressed as a percentage of vehicle control. (See Example 13).

Figure 5 is a generic synthetic scheme for the preparation of Compounds li-XII. (See Example 1).

Figure 6 is a synthetic scheme for the preparation of Compound IV. (See Example 2).

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Figure 7 is a synthetic schematic for -4a.-preparation .dsL Compound VI. (See Example 3).

Figure 8 is a synthetic schematic for the preparation of Compounds IX and X. (See Example 5).

Figure 9 shows testosterone levels in intact rats treated with Compound IV after 24 hours, 72 hours and 168 hours at doses of 3 mg / kg, 10 mg / kg, and 300 mg / kg. (See Examples

9).

Figures 10A to 10F show LH levels (Figure 10A), FSH levels (Figure 10B), testosterone levels (Figure 10C), prostate weight levels (Figure 10D), seminal vesicle weight levels (Figure 10E) , and weight of levator ani (Figure 10F) from intact and orchiectomized (ORX) treated rats at doses of 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 10 mg / kg, and 30 mg / kg of Compound IV. 'denotes P> 0.05 against vehicle controls. ° denotes P <0.05 against ORX vehicle controls. BLOQ values are graphically represented at the 0.08 ng / ml quantification limit. (See Example 9).

Figures 11A and 11 B show prostate size in intact and ORX rats administering Compound IV (Figure 11 A) and DES (Figure 11 B) at different doses. (See Example 15).

Figures 12A to 12C show differences between DES and Compound IV; DES cross-reacts with the glucocortlcolde receptor (GR) while Compound IV does not (Figure 12A); DES cross-reacts with the receptor

MEXICAN INSTITUTE ΠΕ LA FROPIF.OAI)

INDUSTRIAL

androgen (AR). It moderately stimulates the action of AR and moderately inhibits (ie it is a partial agonist / antagonist) while Compound IV does not (Figure 12B); DES abrogates transactivation of the estrogen-related receptor (ERR), while Compound IV does not (Figure 12C) (See Example 15).

Figure 13 shows the effect of Compound IV on attenuation of hot flashes in a morphine withdrawal model at doses of 5 mg / kg, 10 mg / kg, 15 mg / kg, and 30 mg / kg. N = 7 animals per group. 17β-Ε2 was used at 5 mg / kg in 100% DMSO. (See Example 14).

Figure 14 shows monkey dose dependent reductions in body weight (kg) (~ 20% at 100 mg / kg) by administering Compound IV for 91 days. No sign of hyper-estrogenicity was observed. (See Example 16).

Figure 15 shows dose-dependent reductions in serum testosterone level (ng / ml) in monkeys after daily oral administration of Compound IV compared to the positive control (LHRH agonist). The dotted line indicates the testosterone level of chemically castrated patients and the dotted line in bold indicates the testosterone level of the surgically castrated monkey. (See Example 16).

Figure 16 shows dose-dependent prostate-specific antigen (PSA) levels (ng / ml) in monkeys by administering Compound IV to a baseline and at day 28. PSA levels were significantly reduced with Compound treatment IV. (See Example 16).

MEXICAN INSTITUTE

M LA FRORIEDAD

INDUSTRIAL

Figure 17 shows the dose-dependent prostate volume using transrectal ultrasound (TRUS) in monkeys compared to the positive control (LHRH agonist), administering Compound IV at week 6. (See Example 16).

Figure 18 shows dose-dependent organ weights (prostate, seminal vesicle, and testes) as a percentage of monkey control on day 90, administering Compound IV (Figure 18A). Prostate weights at necropsy at week 13 after daily oral administration of Compound IV (Figure 18B). (See Example, 16).

Figure 19 shows average total dose-dependent testosterone levels (nmoles / l) in humans over a period of 1-11 days administering Compound IV (100mg, 300mg, 600mg, and 1000mg). (See Example 17).

Figure 20 shows average dose-dependent LH (IU / I) levels in humans over a period of 1-10 days administering Compound IV (100mg, 300mg, 600mg, and 1000mg). (See Example 1 7).

Figure 21 shows average dose-dependent free testosterone levels (pg / ml) in humans over a period of 1-10 days administering Compound IV (100mg, 300mg, 600mg, and 1000mg). (See Example 17).

Figure 22 shows dose-dependent mean PSA levels (pg / l) in humans over a period of 1-10 days administering Compound IV (100mg, 300mg, 600mg, and 1000

MEXICAN INSTITUTE / ΓΟΕ THE PROPERTY

INDUSTRIAL

mg). (See Example 17). .......-

Figure 23 shows dose-dependent mean testosterone levels (mg / ml) in intact rats after 14 days of recovery from Compound IV administration. 'denotes P <

0.05 against intact controls. (See Example 10).

It will be appreciated that for simplicity and clarity of illustration, the elements shown in the figures have not necessarily been drawn to scale. For example, the dimensions of some of the elements may be exaggerated relative to other elements for clarity. Furthermore, when deemed appropriate, reference numbers may be repeated between the figures to indicate corresponding or analogous elements.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

In the following description, numerous specific details are set forth in order to provide a complete understanding of the invention. However, it will be understood by those skilled in the art that the present invention can be practiced without these specific details. In other cases, well known methods, procedures and components have not been described in detail in order not to obscure the present invention.

In one embodiment, the compounds, as described herein, and / or compositions comprising the same can be used to decrease serum total testosterone levels in a subject

male.

In one embodiment, the compounds, as described herein, and / or compositions comprising the same can be used to decrease serum total testosterone levels in a male subject, where decrease in serum total testosterone occurs through a reduction in serum luteinizing hormone (LH) levels.

In one embodiment, the compounds, as described herein, and / or compositions comprising the same can be used to decrease serum total testosterone levels in a male subject, where the decrease in serum total testosterone is independent of a reduction in serum luteinizing hormone (LH) levels.

(D

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY where:

Y is C (O) or CH2;

R1( R2 they are independently hydrogen, halogen, hydroxyl, alkoxy, cyano, nitro, CF3, N (R)2, sulfonamide, SO2R, alkyl, haloalkyl, arite, O-Alk-heterocycle, wherein the heterocycle is a 3-7 membered substituted or unsubstituted heterocyclic ring, optionally aromatic;

R3, R4 they are independently hydrogen, halogen, hydroxy I qui I or, hydroxyl, alkoxy, cyano, nitro, CF3, NHCOR, N (R)2, sulfonamide, SO2R, alkyl, haloalkyl, aryl, or protected hydroxyl;

R is alkyl, hydrogen, haloalkyl, di haloalkyl, tri ha I or Iq ui I o, CH2F, CHF2, CF3, CF2CF3, aryl, phenyl, halogen, alkenyl, CN, NO2, or OH;

R5 and R6 they are independently hydrogen, halogen, phenyl, an alkyl group of 1 to 6 carbon atoms, a 3-7 membered cycloalkyl, a 3-7 membered heterocycle, a 5-7 membered aryl; or R5 and R6 they form a 3-7 membered ring with the nitrogen atom;

j and k are independently 1-4;

Alq is 1-7 carbon linear alkyl, 1-7 carbon branched alkyl, or 3-8 carbon cyclic alkyl.

In additional embodiments of the methods described herein, the compound of Formula I is represented by formula IA:

where Ri, R2, R3, R4, j and k are as defined for the Formula

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DE LA TOOMEIM D <»53« ;. INDUSTRIAL hydrate or any combination thereof, represented by the structure of formula III:

(IU)

(IV)

In one embodiment, the invention provides a method of lowering serum total testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a compound or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate, or any combination. thereof, represented by the structure of formula V:

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(V)

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY pharmaceutically acceptable, pharmaceutical product, polymorph, 'hydrate or any combination thereof, represented by the structure of the formula Vil:

(vn)

In one embodiment, this invention provides a method of lowering serum total testosterone levels in a subject /

male, comprising administering a therapeutically effective amount of a compound or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate, or any combination thereof, represented by the structure of formula VIII:

(VUI)

MEXICAN INSTITUTE AND say the moniDAO

INDUSTRIAL hydrate or any combination of the structures of formula IX:

themselves, represented by the

(X)

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structure of formula XI;

HO

F

OH (XI)

HO

OH (ΧΠ)

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INDUSTRIAL prostate. In another embodiment, total serum testosterone is decreased below about 100 ng / dL. In another embodiment, total serum testosterone is decreased below about 50 ng / dL. In another embodiment, the total serum testosterone concentration is decreased below about 25 ng / dL.

In one embodiment, this invention provides a method of lowering serum total testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a compound of the formulas ΙΑ, l-XII or its isomer, pharmaceutically acceptable salt, product pharmaceutical, polymorph, hydrate or any combination thereof, where the decrease in total serum testosterone occurs due to a reduction in serum luteinizing hormone (LH) levels. In another embodiment, the male subject has prostate cancer. In another embodiment, total serum testosterone is decreased below about 100 ng / dL. In another embodiment, total serum testosterone is decreased below about 50 ng / dL. In another embodiment, the total serum testosterone concentration is decreased below about 25 ng / dL.

In one embodiment, this invention provides a method of lowering serum free testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a compound of formulas IA, l-XII

MEXICAN INSTITUTE

ΠΕ THE MONEDAD

INDUSTRIAL

or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate, or any combination thereof, wherein the decrease in serum free testosterone occurs by a reduction in serum luteinizing hormone (LH) levels. In another modality; the male subject has prostate cancer.

In one embodiment, this invention provides a method of lowering serum total testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a compound of formulas IA, I-XII or its isomer, pharmaceutically acceptable salt, product pharmaceutical, polymorph, hydrate or any combination thereof, wherein the decrease in total serum testosterone is independent of a reduction in serum luteinizing hormone (LH) levels. In another embodiment, the male subject has prostate cancer. In another embodiment, total serum testosterone is decreased below about 100 ng / dL. In another embodiment, total serum testosterone is decreased below about 50 ng / dL. In another embodiment, the total serum testosterone concentration is decreased below about 25 ng / dL.

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MEXICAN INSTITUTE OF '.A PROPERTY

INDUSTRIAL

or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate, or any combination thereof, wherein the decrease in serum free testosterone levels is independent of a reduction in serum luteinizing hormone (LH) levels. In another embodiment, the male subject has prostate cancer.

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

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

In another embodiment, serum testosterone concentrations are reversible after treatment with Compound IV according to Figure 23 and Example 10.

In the modality, total serum testosterone is decreased below about 50 ng / dL. In another embodiment, the total serum testosterone concentration is decreased below about 25 ng / dL. In another embodiment, total serum testosterone is decreased below about 75 ng / dL. In another embodiment, total serum testosterone is decreased to approximately 75 ng / dL-100 ng / dL. In another embodiment, total serum testosterone is decreased to approximately 50 ng / dL-75 ng / dL. In another embodiment, total serum testosterone is decreased to approximately 40 ng / dL-50 ng / dL. In another embodiment, the total serum testosterone concentration is decreased to approximately between 25 ng / dL-50 ng / dL. In another embodiment, total serum testosterone is decreased to approximately 40 ng / dL-60 ng / dL.

Testosterone can be measured as "Udder" levels (ie, bioavailable or unbound) or as "total" (including the percentage that is bound or unavailable protein) in serum. Men, without prostate cancer, older than 40 years of age demonstrate low testosterone levels having a total testosterone level of less than 250 ng / dL (<8.7 nmol / l) or a level of free testosterone less than 0.75 ng / dL (<0.03 nmol / l).

In one embodiment, the methods of this invention provide a method of decreasing serum total and / or free testosterone levels independent of reduction of luteinizing hormone (LH) levels or reduction of LH levels in a subject.

male who has prostate cancer. In another modality, the 'changes in testosterone levels should be a reduction in level before treatment. In another embodiment, the serum total testosterone level is decreased below 100 ng / dL. In another embodiment, the serum total testosterone level is decreased below 50 ng / dL. In another embodiment, the serum total testosterone level is decreased below 25 ng / dL. In another embodiment, the level of free testosterone is decreased below 2 ng / dL. In another embodiment, the level of free testosterone is decreased below 1 ng / dL. In another embodiment, the level of free testosterone is decreased below 0.5 ng / dL. In another embodiment, the level of free testosterone is decreased below 0.25 ng / dL.

Methods for determining serum free testosterone levels and serum total testosterone levels include checking testosterone levels during the course of the treatment period through a blood test. Total testosterone is a combination of circulating testosterone bound to carrier proteins (albumin, SHBG, transcortin, transferrin) and the free / unbound hormone. Total testosterone levels can be affected by various factors including the level of protein in the blood that carries the hormone in the body, age, obesity and interference associated with commonly used test methods.

Available methods to measure free testosterone (FT) can be complex (balance dialysis or free testosterone

IMPI

MEXICAN INSTITUTE

OF THE PROPERTY

INDUSTRIAL calculated (CFT)) or simple (the commercial FT-Coat-ACount equipment) using an analog tracker. In another embodiment, measurement of serum total testosterone and free testosterone levels can be accomplished through simultaneous measurement of total testosterone and SHBG (eg, Irma-Count, DPC) and then a calculated free testosterone (CFT). In another embodiment, the measurement of total testosterone and free testosterone is according to the knowledge of one skilled in the art.

In one embodiment, this invention provides a method of lowering serum total testosterone levels or serum free testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a combination of one or more forms of ADT and a compound. of formula IA, l-XII or its isomer, pharmaceutically acceptable salt. Polymorph, hydrate, or any combination thereof. In another embodiment, the decrease in total or free serum testosterone occurs by a reduction in the level of serum luteinizing hormone (LH). In another embodiment, the decrease in serum total or free testosterone levels is independent of a reduction in serum luteinizing hormone levels.

The methods of this invention comprise administering a combination of other forms of ADT and a compound of this invention. In one embodiment, other forms of ADT include an LHRH agonist. In another embodiment, the LHRH agonist includes leuprolide acetate (Lupron®) (US 5,480,656; US 5,575,987;

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OL THE INDUSTRIAL ROPE

5,631,020; 5,643,607; 5,716,640; 5,814,342; 6,036,976, which are incorporated herein for reference) or gcserelin acetate (Zoladex®) (US 7.1 1 8,552; 7,220,247; 7,500,964, which are incorporated herein for reference). In one embodiment, other forms of ADT include an LHRH antagonist. In another embodiment, the LHRH antagonist includes degarelix. In one embodiment, other forms of ADT include anti-androgens. In another embodiment, anti-androgens include bicalutamide, flutamide, finasteride, dutasteride, MDV3100, nilutamide, chlormadinone, or any combination thereof.

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

In one embodiment, this invention provides a method of decreasing serum total testosterone levels and / or free testosterone levels through reduction of luteinizing hormone (LH) levels or independent of reduction of hormone levels luteinizing in a male subject with prostate cancer for the purpose of producing an androgen deprivation therapy (ADT) comprising administering an amount

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INSTITUTO MEXICANO industrial 'therapeutically effective of a compound of the formula ΙΑ, 1-XII. In another embodiment, the compound is Compound IV.

In another embodiment, this invention provides a method for androgen deprivation therapy (ADT) in a subject, which comprises administering a therapeutically effective amount of a compound of formula IA, l-Xlt or its isomer, pharmaceutically acceptable salt, product pharmaceutical, polymorph, hydrate or any combination thereof. In another embodiment, said subject has prostate cancer. In another embodiment, the compound is Compound IV.

In another embodiment, ADT is used to treat prostate cancer, to slow the progression of prostate cancer, or to prevent and / or treat recurrence of prostate cancer.

In another embodiment, this invention provides a method of treating prostate cancer, for delaying the progression of prostate cancer, for preventing and / or treating recurrence of prostate cancer, which comprises administering a compound of this invention. In another embodiment, administration of a compound of this invention may be in combination with L.HRH analogs, reversible anti-androgens (such as blcalutamide or flutamide), anti-estrogens, anti-cancer drugs, 5-alpha reductase inhibitors , Aromatase inhibitors, progestins, selective androgen receptor modulators (SARMS), or agents that act through other nuclear hormone receptors.

In one embodiment, the present invention provides a method

Mexican Institute of Industrial Property

for treating prostate cancer and reducing serum total testosterone and / or serum free testosterone levels, by reducing LH levels or LH levels reduction independent, which comprises administering a compound of formula IA, l- XII. In another embodiment, administer Compound IV.

Androgen deprivation therapy not only lowers testosterone levels, but estrogen levels are also lower as estrogen is derived from aromatization of testosterone. Androgen Deprivation Therapy-Induced Deficiency Causes Significant Side Effects, Including Hot Flashes, Gynecomastia, and Masstagia, Bone Loss, Reductions in Bone Quality and Strength, Osteoporosis, Osteopenia, and Life-threatening Fractures, Changes adverse lipids, and superior cardiovascular disease and myocardial infarction, loss of livid, impotence, loss of muscle mass (sarcopenia), fatigue, cognitive dysfunction, and depression and other mood swings.

In other embodiments, the present invention provides a method of treating any disease, disorder, or symptom associated with ADT. In other embodiments, the present invention provides a method of treating any disease, disorder, or symptom associated with testosterone deprivation. Each disease, disorder, or symptom represents a separate embodiment of the present invention.

In one embodiment, this invention provides a method for

lowering serum total testosterone levels in a male subject, which comprises administering a therapeutically effective amount of a compound of formula IA, l-XII or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate or any combination of the same, wherein said administration of said compounds of formula IA, l-XII or its isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate or any combination thereof, prevents, suppresses, reduces the incidence, inhibits or tries not to cause side effects associated with androgen deprivation therapy (ADT), where said subject has prostate cancer. In another embodiment, the decrease in serum total or free testosterone levels is through reduction of LH levels or independent of reduction in LH levels.

In one embodiment, administration of the compounds of this invention suppresses, reduces the incidence, inhibits, or attempts to prevent typical side effects from occurring with traditional androgen deprivation therapy (ADT). In another embodiment, the subject has prostate cancer. Said prevention and / or reduction of side effects is relative to the placebo or control group. In one modality, the typical side effects associated with traditional androgen deprivation therapy (ADT) include hot flashes, gynecomastia, reduced bone mineral density, and increased bone fracture. In another embodiment, administration of the compounds of this invention prevents hot flashes from occurring, such as

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OF THE PROPERTY

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They could occur when using traditional forms of androgen deprivation (ADT) therapy. In another embodiment, administration of the compounds of this invention prevents gynecomastia from occurring, as might occur when using traditional forms of androgen deprivation therapy (ADT). In another embodiment, administration of the compounds of this invention prevents reduced bone mineral density (BMD) from occurring, as might occur when using traditional forms of androgen deprivation therapy (ADT). In another embodiment, administration of the compounds of this invention prevents increased bone fractures from occurring, as might occur when using traditional forms of androgen deprivation therapy (ADT). In another modality, increased bone fractures are pathological fractures, non-traumatic fractures, vertebral fracture, non-vertebral fractures, new morphometric fractures, or a combination thereof.

In one modality, the term "traditional androgen deprivation therapy" is directed at orchidectomy (surgical castration), where the surgeon removes the testicles. In another embodiment, the term "traditional androgen deprivation therapy" is directed at the administration of luteinizing hormone-releasing hormone (LHRH) analogs; These drugs reduce the amount of testosterone made by the testicles. Examples of LHRH analogues available in the United States include leuprolide (Lupron, Viadur, Ellgard), goserelin (Zoladex), triptorelin (Trelstar), and

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MEXICAN INSTITUTE iL · ** »» ** ^ * .. *!

OWNED O »« Jr ^ ÚU¿S¿F INDUSTRIAL histrelina (Vantas). In another embodiment, the term "traditional androgen deprivation therapy" is directed at the administration of antiandrogens: anti-androgens block the body's ability to use any androgen. Even after an orchidectomy or during treatment with LHRH analogues, a small amount of androgen is still made by the adrenal glands. Examples of anti-androgen drug include flutamide (Eulexin), bicalutamide (Casodex), and nilutamide (Nllandron). In another embodiment, the term "traditional androgen deprivation therapy" is intended for administration of luteinizing hormone-releasing hormone (LHRH) antagonists such as Abarelix (Plenaxls); Degarelix (Firmagon) is a new LHRH antagonist that was approved for use by the FDA in 2008 to treat advanced prostate cancer. In another embodiment, the term "traditional androgen deprivation therapy" is directed at the administration of 5a-reductase inhibitors such as finasteride (Prosear) and dutasterlda (Avodart): 5a-reductase inhibitors block the body's ability to convert Testosterone to the most active androgen, 5a-dihydrotestosternoa (DHT). IN another embodiment, the term "traditional androgen deprivation therapy" is directed at the administration of testosterone biosynthesis inhibitors such as ketoconazole (Nizoral). In another embodiment, the term "traditional androgen deprivation therapy" is directed at the administration of estrogens such as diethylstilbestrol or 1 7βestradiol.

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INOUJTitlAL

In one embodiment, the term "hot flashes" refers to a sudden sensation of heat in the upper or whole body, redness of the face and neck, red rashes appearing on the chest, back and arms, excessive sweating, chills, etc. .

In one embodiment, the term "gynecomastia" refers to a benign enlargement of the male chest that results from a proliferation of the glandular component of the chest, which may or may not be associated with pain. Gynecomastia is clinically defined by the presence of an elastic or firm mass that extends concentrically from the nipples. The condition known as pseudoginecomastia, or lipomastia, is characterized by fat deposition without glandular proliferation. Although gynecomastia is t

usually bilateral, may be unilateral.

In one embodiment, the methods of this invention are directed to treating men with prostate cancer or advanced prostate cancer through reduced testosterone without causing bone loss and hot flashes. In another embodiment, the methods of this invention make use of compounds IA-1-XII, where the compounds have the potential to reduce testosterone, a primary stimulus for prostate cancer, without causing certain side effects such as bone loss and hot flashes that are common with current androgen deprivation therapy (ADT) for prostate cancer.

In another embodiment, Table 8 (Example 11) below, demonstrates testosterone reduction without causing bone loss by

administer Compound IV.

In one embodiment, the methods of this invention are directed at reducing testosterone levels which further treats patients with advanced prostate cancer by administering a compound of formula IA, l-XII. In one embodiment, the methods of this invention are directed at reducing testosterone levels which further suppress, reduce incidence, reduce severity, or inhibit advanced prostate cancer by administering a compound of formula IA, l-XII. In one embodiment, the methods of this Invention are directed at reducing testosterone levels which also provides a palliative treatment for advanced prostate cancer by administering a compound of formula IA, l-XII.

In one embodiment, the methods of this invention are directed to the treatment of advanced prostate cancer. In one embodiment, the methods of this invention are directed at suppressing, reducing incidence, reducing severity, or inhibiting advanced prostate cancer. In one embodiment, the methods of this invention are directed to the palliative treatment of advanced prostate cancer. In> one embodiment, the methods of this invention make use of compounds IA, l-XII. In one embodiment, this invention is directed to the treatment of advanced prostate cancer. In one embodiment, this invention is directed at suppressing advanced prostate cancer. In one embodiment, this invention is directed at reducing the incidence of advanced prostate cancer. In one embodiment, this invention is

aimed at reducing the severity of advanced prostate cancer. In one embodiment, this invention is directed at inhibiting advanced prostate cancer.

The term "advanced prostate cancer" refers to metastatic cancer that originated in the prostate, and has become extensively metastatic beyond the prostate, such as the surrounding tissues, to include seminal vesicles, pelvic lymph nodes, or bone. , or to other parts of the body. Prostate cancer pathologies are classified with a Gleason score of 1 to 5 in order of increasing malignancy. In another modality, patients at significant risk for progressive disease and / or death from prostate cancer should be included in the definition and that any patient with cancer outside the prostate capsule with disease stages as low as 11B clearly has cancer. " advanced".

In one embodiment, the methods provided herein and / or using the compounds provided herein are effective in providing feedback on the hypothalamic-pituitary-testlcular axis (HPT axis).

Feedback refers to the ability of a substance produced in one organ or tissue to regulate the activity of another organ or tissue that affects its own activity. In one modality, feedback on the hlpotálamo-pitultaria-testicular axis (HPT axis) results in the reduction of LH levels. In one embodiment, re-growth on the axis of the hypotá I amo - ρ ¡tu it ar ¡a testicular (HPT axis) results in reduced levels of

total serum testosterone. In one embodiment, feedback-eTT to the hypothalamic-pituitary-testicular axis (HPT axis) results in reduced serum free testosterone levels. In one embodiment, feedback on the hypothalamic-pituitary arctic axis (HPT axis) results in reduced androgen levels in serum, tissue, or tumor.

The hypothalamic-pituitary-testicular axis (HPT axis) refers to the endocrine physiological system that regulates hormone levels in the hypothalamus, pituitary gland, and testes. LHRH (luteinizing hormone releasing hormone) is released by the hypothalamus and stimulates the pituitary to synthesize and secrete LH and FSH (gonadotropins). LH and FSH then act on the testicles to stimulate the production of testosterone and sperm. Testosterone then has a direct negative feedback effect on hypothalamic LHRH secretion and a negative and indirect feedback effect on pituitary LH and FSH production. Serum estrogens, androgens, and proteins (eg, inhibin) also have a negative effect on LHRH secretion and LH and FSH secretion.

The pituitary gland is a gland that controls the level of testosterone in the body. When the testosterone level is low, the pituitary gland releases luteinizing hormone (LH). This hormone induces the testicles to make more testosterone. Testosterone level increases during puberty. Testosterone level is highest at the age of about 20-40 years, and

then it gradually decreases in larger men. Women have a much smaller amount of testosterone in their bodies compared to men. But testosterone plays an important role throughout the body in both men and women. It affects the brain, bones and muscle mass, fat distribution, the vascular system, energy levels, genital tissues, and sexual function. Most testosterone in the blood is bound to a protein called sex hormone binding globulin (SHBG) or to another serum protein called albumin. Testosterone that is not bound or fixed (or free) can also be clinically determined.

In another embodiment, the decrease in serum total testosterone or serum free testosterone levels independent of a reduction in serum luteinizing hormone levels is due to an increase in sex hormone binding globulin (SHBG). In another embodiment, the decrease in free testosterone levels independent of a reduction in luteinizing hormone levels is due to the increase in sex hormone binding globulin (SHBG). In another embodiment, the decrease in serum total or free serum testosterone levels independent of a reduction or reduction in luteinizing hormone levels is due to inhibition of testosterone production or secretion by Leydig cells in the testes. . In another embodiment, lowering serum total or free serum testosterone levels independent of one

37__ reduction a reduction in luteinizing hormone levels is due to reduced adrenal steroidogenesis.

In one embodiment, the compounds, as described herein, and / or compositions comprising the same can be used for 1st reduction of luteinizing hormone (LH) levels. In another embodiment, the compounds and / or compositions of this invention can be used to reduce endogenous sex hormones.

Members of the hydroxysteroid dehydrogenase (HSD) family are involved in the conversion of circulating spheroids. 17p-HSD5 converts androstenedione to testosterone and estrone to estradiol. In addition, it is also involved in the synthesis of prostaglandin. In one embodiment, the compounds of this invention specifically inhibit HSD by inhibiting 17β-hydroxysteroid dehydrogenase 5 (17p-HSD5). Such inhibition may be useful in ADT, avoiding peripheral / extragonadal synthesis of testosterone that may escape control of the HPT axis and cause incomplete reduction of serum total or free testosterone or allow locally elevated intracellular testosterone levels, any of the which can be harmful in the ADT.

Androgen Deprivation Therapy (ADT) accomplished by LHRH agonist therapy, i.e., administering luteinizing hormone-releasing hormone (LHRH) agonists or analogues thereof, results in initial stimulation of gonadotropin release of the pituitary and testosterone production of the testicles (called "flare reaction"),

I JM [P í Mexican institute of INDUSTRIAL property followed by decreased release of gonad tTüjptrrar -'- and * - decreased levels of both testosterone and estrogen.

The flare reaction ”caused by LHRH agonist therapy has a negative impact on prostate cancer treatment, due to increased androgen / testosterone levels. Furthermore, LHRJ therapy has been associated with an increased risk of diabetes and cardiovascular disease (Smith (2008) Current Prostate Reports. 6: 149-154).

In an effort to overcome the flare effects of LHRH therapy, anti-androgen monotherapy (bicalutamide, flutamide, chlormadinone), combined aspects of LHRH / anti-androgen therapy, and LHRH (degarelix) antagonists (Suzuki) have been suggested. et al., (2008) Int. J. Clin. Oncol. 13: 401-410; Sharifi, N. et al., (2005) JAMA. 294 (2): 238-244). Anti-androgen monotherapy does not reduce androgen levels in a subject. Anti-androgen monotherapy, bicalutamide, was shown to be less effective than ADT in patients with prostate cancer with bone metastases. Furthermore, the adverse effects observed with bicalutamide therapy include breast tenderness and breast enlargement (Gynecomastia and Mastodynia). (Suzuki et al., Ibid). An additional risk with anti-androgen therapy includes elevated levels of liver transaminases (Sharifi et al. Ibid).

In one embodiment, the present invention provides a reduction in LH levels and thus a reduction in serum total testosterone and / or free testosterone levels

ΙΜΡΙ

MEXICAN INSTITUTE

Dt THE PROPERTY

2Q INDUSTRIAL in serum, without producing the "flare" effect, and while "overcoming the adverse effects associated with estrogen deficiency caused by testosterone reduction using ADT methods. The methods / uses of the compounds herein provide tissue selective estrogen activities that provide maintenance to bone tissue (agonistic effect on bone tissue), reduced thrombotic potential and / or hot flashes and / or less or neutral effects on tissue of the chest than estradiol or diethylstilbestrol.

In one embodiment, Compound IV exhibits agonistic but non-antagonistic effects (Examples 6 and 7) whereby Compound IV cannot cause an increase in gonadotropins and testosterone.

In one embodiment, Compound IV exhibits agonist activity (Examples 8-11) demonstrating a robust pharmacological response to the reduction of hormones, testosterone, and total serum androgens.

In one embodiment, the methods provided herein using the compounds and / or compositions provided herein, are effective in reducing or eliminating the bone resorption effects caused by LH reduction using traditional forms of ADT. In one embodiment, the methods provided herein and / or using the compositions provided herein are effective in reducing or eliminating bone resorption effects caused by reduced testosterone levels using traditional forms of ADT. In one embodiment, the methods provided here using the

IMPIOUS

WniTO MEXICANO ...

OF PROPERTY .3,

INDUSTRIAL compositions provided herein are effective in reducing or eliminating the bone resorption effects caused by the reduction of estrogen as a result of the reduction of the LH level. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein, avoid the bone resorption effects associated with lowering the LH level using traditional forms of ADT. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein, prevent bone loss associated with the reduction of endogenous LH, testosterone and / or estradiol using traditional forms of ADT. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein, increase bone mass density (BMD) while providing a reduction in the LH level. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein, increase the percentage of

X bone volume while providing the reduction of the level of endogenous LH, testosterone and / or estradiol.

In some embodiments, this invention provides a method of preventing and / or reducing thromboembolism by administering a compound of this invention or its isomer, pharmaceutical, polymorph, hydrate, or any combination thereof.

In one embodiment, the methods provided herein using the compounds and / or compositions provided herein, are effective on breast tissue. In one embodiment, the methods provided herein using the compounds and / or compositions provided herein,

They provide the LH level reduction while avoiding gynecomastia associated with the LH level reduction achieved through traditional ADT.

In one embodiment, Example 13 describes special toxicity studies, where in vitro studies with human platelets showed that Compound IV had much less pro-coagulant activity than DES. Thus, Compound IV, an ER-selective agonist, must provide the benefits of DES for prostate cancer with a lower risk of thrombotic events than DES, and also provide the benefits of an LHRH agonist or antagonist without causing bone loss. , hot flashes, or adverse lipid profiles.

Diethylstilbestrol (DES) therapy alone or in combination with another ADT showed that DES prevented bone resorption in patients with prostate cancer. Although the use of DES has been promoted as a therapy for prostate cancer, the effects of DES on angiogenesis and malignancy are believed to be mediated by DES metabolites and are not believed to act via the estrogen receptor. Furthermore, the dose levels of DES administered for therapeutic uses have numerous adverse side effects including vascular disease, cardiovascular morbidity, thrombotic toxicity, gynecomastia, erectile dysfunction, and reduced livid (Scherr and Pitts, ibid and Prestí, JC Jr. (1996) JAMA 275 (15): 1 1 53-6).

In one embodiment, the present invention overcomes the negative side effects of LHRH agonist or antagonist therapy, alone

or in combination with anti-androgens or DES. In other modalities, the methods of the present invention provide androgen deprivation therapy without adverse estrogen deprivation side effects, such as bone-related conditions, and without adverse estrogen stimulation side effects, such as gynecomastia. In another embodiment, the methods of the present invention provide a reduction in LH levels and thus a reduction in serum levels of total and / or free testosterone, without producing the "flare" effect, while Outweigh the adverse effects associated with estrogen deficiency caused by LH reduction and while outweigh the adverse effects associated with an increase in estrogen agonist generated! observed in DES therapy. The methods / uses of the compounds of the invention provide tissue selective estrogen activities thus providing bone tissue maintenance (agonist effect on bone tissue), reduced thrombotic potential and neutral effects on breast tissue.

The anti-estrogenic effects of traditional selective estrogen receptor modulators (SERMs) such as tamoxifen, toremifene, and raloxifene at the hypothalamic level result in increased gonadotropin levels or increased LH levels in men, thus resulting potentially an increase in serum testosterone levels. (Tsouri et al., 2008, Fertility and Sterility doi: 10,1016), In contrast, the methods of this invention provide LH reduction in a subject

male, comprise administering a compound of formula ΙΑ, IXII.

Additional Modalities for the Compound of Formula I:

In one embodiment of the methods of this invention, Y of the compound of formula I is C (O). In another modality, Rt and R2 of the compound of formula I or IA are independently O-Alq-NR5R6 or O-Alq-heterocycle. In another embodiment, the Alq of said O-Alqheterocycle, O-Alq-NR5R6, -Alq-heterocycle, and Alq-NR5R5, as described herein above, are linear 1-7 carbon alkyl, branched 1-7 carbon alkyl, or cyclic 3-8 carbon alkyl. In another embodiment, the alkyl is ethylene (-CH2CH2-). In another embodiment, the Alq is methylene (-CH2-). In another embodiment, the Alq is propylene (-CH2CH2CH2-). In another embodiment, the Alq is 2-methylpropylene (-CH2CH (CH3) CH2-).

In one embodiment of the methods of this invention R1 of the compound of formula I or IA is in the para position. In one embodiment of the methods of this invention, Rt and R2 of the compound of formula I or IA are different. In another embodiment of the methods of this invention, R ·, and R2 of the compound of formula I or IA are the same. In another embodiment of the methods of this invention, Ri of the compound of formula I or IA is

In another embodiment of the methods of this invention, Rt of the compound of formula I or IA is hydroxyl. In another embodiment of the methods, Rt del

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MEXICAN INSTITUTE W THE INDUSTRIAL PROPERTY

compound of formula I or IA is alkoxy. In another embodiment of the methods, R, and R2 they are independently hydrogen, halogen, hydroxyl, alkoxy, cyano, nitro, CF3, N (R)2, sulfonamide, SO2R, alkyl, haloalkyl, aryl, O-Alq-NR5R6, or O-Alq-heterocycle, wherein the heterocycle is a heterocyclic ring substituted or unsubstituted with

3-7 members, optionally aromatic. In another embodiment of the methods, Ri and R2 of the compound of formula I or IA are independently hydrogen, halogen, hydroxyl, alkoxy, cyano, nitro, CF3, N (R)2, sulfonamide, SO2R, alkyl, haloalkyl, aryl, 0Alq-NR5R6, or O-Alq-heterocycle, wherein the heterocycle is a 3-7 membered substituted or unsubstituted heterocyclic ring, optionally aromatic. In another embodiment of the methods, R2 of the compound of formula I or IA is halogen. In another embodiment of the methods, R2 of the compound of formula I or IA is F. In another embodiment of the methods, R2 of the compound of formula I is Cl. In another embodiment of the methods, R2 of the compound of formula I or IA is Br. In another embodiment of the methods, R2 of the compound of formula I or IA is I. In another embodiment of the methods, R2 of the compound of formula I or IA is hydroxyl. In another modality of the methods, R-ι and / or R2 is CF3. In another modality, Rj and / or R2 is CH3. In another modality, R-ι and / or R2 it is halogen. In another modality, Rj and / or R2 is F. In another modality, Ri and / or R2 is Cl. In another embodiment, R-, and / or R2 is Br. In another modality, Rj and / or R2 is I. In another modality, R2 of the compound of formula I is in the para position.

In one embodiment of the methods of this invention, R3 and R4

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JCANO OI INSTITUTE INDUSTRIAL AROPIEDAD

of the compound of formula I or IA are the same. In another embodiment of the methods of this invention, R3 and R4 of the compound of formula I or IA are different. In another embodiment of the methods, j and k of y

compound of formula I or IA are independently 1. In another embodiment of the methods, R3 and R4 of the compound of formula I or IA are independently halogen, haloalkyl, hydroxyl, or alkyl. In another embodiment of the methods, R3 and R4 of the compound of formula I or IA are independently F. In another embodiment of the methods, R3 and R4 of the compound of formula I or IA are independently Br. In another embodiment of the methods, R3 and R4 of the compound of formula I or IA are independently Cl. In another embodiment of the methods, R4 is in the position to. In another embodiment of the methods, R3 is in the ortho position. In another embodiment of the methods, R3 is in the meta position. In another embodiment of the methods, R3 and / or R4 is CF3. In another embodiment of the methods, R3 me

R4 is CH3.

In one embodiment of the methods of this Invention, Rs and Rand of the compound of formula I or IA form a 3-7 membered ring with the nitrogen atom. In another embodiment, the ring is a saturated or unsaturated ring. In another embodiment, the ring is a substituted or unsubstituted ring. In another embodiment of the methods of this invention, R5 and R6 of the compound of the formula I or IA form a ring of plperlin with nitrogen. In another embodiment of the methods, R5 and R6 of the compound of formula I or IA form a pyrazine ring with nitrogen. In another embodiment of the methods, R5 and

R6 of the compound of formula I or IA form n eleven ? piperazine with nitrogen. In another embodiment of the methods, R5 and Rs of the compound of formula I or IA form a morpholine ring with nitrogen. In another embodiment of the methods, R5 and R6 of the compound of formula I or IA form a pyrrole ring with nitrogen. In another embodiment of the methods, R5 and R6 of the compound of formula I or IA form a pyrrolidine. In another embodiment of the methods, R5 and R6 of the compound of formula I or IA form a pyridine ring with nitrogen. In another embodiment, the ring is substituted by halogen, alkyl, alkoxy, alkylene, hydroxyl, cyano, nitro, amino, amide, COOH, or an aldehyde.

In another embodiment of the methods of this invention, R1 of the compound of formula I or IA and R2 of the compound of formula I or IA are independently O-Alq-heterocycle or OCH2-CH2heterocycle. In another embodiment, the term "heterocycle" group refers, in one embodiment, to a ring structure that further comprises carbon, sulfur, oxygen, nitrogen, or any combination thereof, as part of the ring. In another embodiment, the heterocycle is a 3-12 membered ring. In another embodiment, the heterocycle is a 6-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 heterocycle group may be unsubstituted or substituted by a halogen, haloalkyl, hydroxyl, alkoxy, carbonyl, amido, alkylamido, dialkylamido, cyano, nitro, CO2H, amino,

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY !.

alkylamino, dialkylamino, carboxyl, uncle, and / or thioalkyl. -In another embodiment, the heterocycle ring may be fused to another saturated or unsaturated cycloalkyl or 3-8 membered heterocyclic ring. In another embodiment, the heterocyclic ring is a saturated ring. In another embodiment, the heterocyclic 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" refers to a non-aromatic monocyclic or polycyclic ring comprising carbon and hydrogen atoms. A cycloalkyl group can have one or more carbon-carbon double bonds in the ring as long as the ring is not presented as aromatic by its presence. Examples of cycloalkyl groups include, but are not limited to, (C3-C7) alkyl groups, such as cyclopropl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, and saturated cyclic and cyclic terpenes and (C3-C7) cycloalkenyl groups, such as cyclopropenyl, cyclobutenyl, cyclopentyl, cyclohexenyl, and cycloheptenyl, and unsaturated cyclic and bicyclic terpenes. A cycloalkyl group can be unsubstituted or substituted by one or two substituents. Preferably, the cycloalkyl group is a monocyclic ring or bicyclic ring.

The term "alkyl" refers, in one embodiment, to a saturated aliphatic hydrocarbon, including straight-chain, branched-chain, and cyclic alkyl groups. In one modality, the group

IMPI

INSn l 'JTO MEKICANt T OF INDUSTRIAL PROPERTY

alkyl has 1-12 carbons. In another modality, the group 31 q írftü'trerve '

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 an alkyl substituted by alkyl side chains of 1 to 5 carbons. In another embodiment, the branched alkyl is an alkyl substituted by haloalkyl side chains of 1 to 5 carbons. The alkyl group can be unsubstituted or substituted by a halogen, haloalkyl, hydroxyl, alkoxy, carbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxyl, uncle and / or thioalkyl.

An "alkenyl" group refers, in another embodiment, to an unsaturated hydrocarbon, including straight-chain, branched-chain, and cyclic groups having one or more double bonds. The alkenyl group can have one double bond, two double bonds, three double bonds, etc. 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 are ethenyl, propenyl, butenyl, cyclohexenyl, etc. the alkenyl group may be unsubstituted or substituted by a halogen, hydroxy, alkoxy carbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxyl, uncle, and / or thioalkyl.

An "aryl" group refers to an aromatic group having i mssyja

IMP

INSTITUTO MEXICANO '/hí7.-.T Dt INDUSTRIAL PROPERTY at least one aromatic carbocyclic group or ^ “φ'τΐ'Ρ'ΰ' Tlüf'eT'CTütctiütr aromatic, which may be unsubstituted or substituted by one or more groups selected from halogen, haloalkyl, hydroxy, alkoxy carbonyl, amido, alkylamldo, dlalkylamido, nitro, amino, alkylamino, dlalkylamino, carboxyl or thio or thioalkyl. Non-limiting examples of aryl rings are phenyl, naphthyl, plranyl, pyrrolllo, pyrazyllo, pyrimidinyl, pyrazolyl, plridinyl, furanyl, thiophenyl, thiazolllo, mldazolyl, isoxazolyl, and the like. In one embodiment, the aryl group is a 4-8 membered ring. In another embodiment, the aryl group is a 4-12 membered ring (s). 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.

An "aldehyde" group refers, in one embodiment, to an alkyl or alkenyl substituted by a formyl group, wherein the alkyl or alkenyl are as defined above. In another embodiment, the aldehyde group is an aryl group, or phenyl substituted by a formyl group, where 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.

A "haloalkyl" group refers, in another embodiment, to an alkyl group as defined above, which is substituted by one or more halogen atoms, eg, F, Cl, Br, or I.

A "hydroxyl" group refers, in another embodiment, to a group

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OF INDUSTRIAL CHALLENGE

- •• -ΐ

Oh It is understood by a person skilled in the art that when R / 7 K2, '™ ü · R3, in the compounds of the present invention, is OR, so R is not OH.

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

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

Referring to protected hydroxyl, in some embodiments, it includes the incorporation of a substituent attached to the oxygen portion of the benzene ring, where the substituent can be easily removed. In some embodiments, phenolic protecting groups may comprise: methyl ether (methoxy), alkyl ether (alkoxy), benzyl ether (Bn), methoxymethyl (MOM) ether, benzoyloxymethyl (BOM) ether, benzyl, carbobenzoxy, methoxyethoxymethyl (MEM) ether, 2 - (t ri m eti I si I il) et ox im et i I (SEM) ether, meti It iom et i I (MTM) ether, phenylthiomethyl (PTM) ether, azidomethyl ether, cyanomethyl ether, 2,2-dichloro-1,1-difluoroethyl ether, 2-chloroethyl ether, 2-bromoethyl ether, tetrahydropyranyl (THP) ether, 1-ethoxyethyl (EE) ether, phenacyl ether, 4bromophenacyl ether, cyclopropylmethyl ether, allyl ether, propargyl ether, isopropyl ether, cyclohexyl ether, t-butyl ether, 2,6-dimethylbenzyl ether, 4-methoxybenzyl ether, o-nitrobenzyl ether, 2,6-d-chlorobenzyl ether, 3,4-dichlorobenzyl ether, 4- (dimethylamino) carbonylbenzyl ether, 4methIsuIfIniIbenciI ether, 4-anthrylmethyl ether, 4-picolyl ether, heptafluorop-tolyl, tetrafluoro-4-pyridyl ether, tr i m i i (TMS) ether, tbutyldimethylsiyl (TBDMS) ether , tb ut ildifeni I si I i I (TBDPS) ether,

ΙΜΡΙ

MEXICAN INSTITUTE Jf,

SAY THE PROPERTY 'Ote.ZCsS_JSV

INDUSTRIAL tri is op ro pi I si I i I (TIPS) ether, arilu formate, classroom acetate. ,, aryl levulinate, arite pivaloate, aryl benzoate, aryl 9-fluorenecarboxylate, aryl methyl carbonate, 1adamantllo carbonate , aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate, aryl benzyl carbonate dimethylphosphnyl ester (Dmp-OAr), d-methylphosphine-oni I ester (Mpt-OAr), diphenylphosphinothionyl ester (Dpt-OAr), aryl methanesulfonate, aryl toluenesulfonate or aryl 2-formylbenzenesulfonate.

In one embodiment, the methods of this invention make use of W, A / -bis (4-hydrophenyl) -4-propylbenzamide (II) or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate or any combination thereof. In another embodiment, the methods of this Invention make use of 4,4 '- (2,3-dimet enό enci I aza nodii I) d if e no I (II) or its isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, or any combination thereof. In another embodiment, the methods of this invention make use of 3-fluoro-A / - (4-fluorophenyl) -4-hydroxy- / V- (4hydroxyphenyl) benzamide (IV) or its Isomer, pharmaceutically salt acceptable, pharmaceutical, polymorph, hydrate or any combination thereof. In another embodiment, the methods of this invention make use of N, N-bis (4-h id roxifeniI) -2,3-dimethi I benzamide (V) or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate or any combination thereof. In another embodiment, the methods of this invention make use

IMPI

MEXICAN INSTITUTE

OF PROPERTY v; ,

INDUSTRIAL de / V, Λ / -b¡s (4-hidroxifeni 1) -2-naftiIamida (VI) or its pharmaceutically acceptable lyüiuerre? —'- sat—, pharmaceutical product, polymorph, hydrate or any combination thereof. In another embodiment, the methods of this invention make use of 3-fluoro-4-hydroxy-A /, A / b¡s (4-hydroxyphenyl) -benzamide (VH) or its isomer, pharmaceutically acceptable sai , pharmaceutical, polymorph, hydrate or any combination thereof. In another embodiment, the methods of this invention use a 4- ((4-fluorophenyl) (4-hydroxybenzyl) amino) phenol (VIII) or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate, or any combination thereof. . In another embodiment, the methods of this invention make use of a 4-f I uo ro-Λ / - (4 - hid ro xi-f eni I) - / V- [4- (2piperidin-1 -¡l-ethoxy ) -phenyl] -2-trifluoromethyl-benzamide (IX) or its isomer, pharmaceutically acceptable sai, pharmaceutical, polymorph, hydrate or any combination thereof. In another embodiment, the methods of this invention make use of a hydrochloride salt of IX (HCI salt of IX) or 4-fluoro- / V- (4hydroxy-phenyl) - / V- [4- (2- p! peridin-1-yl-ethoxy) -phenyl] -2-trifluoromethibenzamide (X) or its isomer, pharmaceutically acceptable salt, pharmaceutical, polymorph, hydrate or any combination thereof. In another embodiment, the methods of this invention make use of a 3-fluoro-4-hydroxy - / \ / - (4-hydroxyphenyl) - / \ / - phenylbenzamide (XI) or its isomer, pharmaceutically acceptable salt, pharmaceutical product, polymorph, hydrate, or any combination thereof. In another embodiment, the methods of this invention make use

of a 3-fluoro- / V, / V-bis- (4-hydroxy-phenyl) -2-methyl-benzamide (XII) or its' isomer, pharmaceutically acceptable salt, pharmaceutical product;

polymorph, hydrate, or any combination thereof.

In one embodiment, the methods of this invention make use of "pharmaceutically acceptable salts" of the compounds, which can be produced, by reacting a compound of this invention with an acid or base.

Suitable pharmaceutically acceptable salts of the amines of the compounds of the methods of this invention can be prepared from an inorganic acid or an organic acid. In one embodiment, examples of inorganic amine salts are bisulfates, borates, bromides, chlorides, hemisulfates, hydrobromates, hydrochlorides, 2-hydroxyethylsulfonates (hydroxyethanesu ifonates), iodates, iodides, isothionates, nitrate, persulfates, phosphate, sulfates, sulfamates, sulfanilates, sulfonic acids (alkylsulfonates, arylsulfonates, halogen-substituted alkylsulfonates, halogen-subtitled arylsulfonates), sulfonates and thiocyanates.

In one embodiment, examples of organic amine salts can be selected from classes of organic aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carbocyclic, and sulfonic acids, examples of which are acetates, arginines, aspartates, ascorbates, adipates, anthranilates, alginates, Alloxy carboxylates, Alloxy substituted carboxylates, Alginate, Benzenesulfonate, Benzoate, Bisulfate, Butyrate, Bicarbonate,

ΙΝΑ-Πτυτο MEXICAN

Dt THE PROPERTY

INDUSTRIAL bitartrates, carboxylates, citrates, camphorrates, camphor sulfonates, cyclohexylsulfamates, cyclopentanpropionates, calcium edetates, camsylates, carbonates, clavulanates, cinnamates, dicarboxylates, dodecyl sulphates, dichlorohydrates, decanoates, enantholates, decanolates, decanolates , formats, fluorides, galacturonates, gluconates, glutamates, glycolate, glucorates, glucoheptanoates, glycerophosphates, gluceptates, g I ico I ¡I a rs ani I at os, glutarates, glutamate, heptanoates, hexanoates, hydroxylealeates, hydroxycarboxylic acids, hexylresorcinates, hydroxybenzoates, hydroxynaphthates, hydrofluorides, lactates, lactobionates, laurates, malates, maleates, methylenebis (beta) , mandelates, mesylates, methanesulfonates, methyl bromides, methyl nitrates, methyl sulfonates, monopotassium maleates, mucates, monocarboxylates, nitrates, naphthalenesulfonates, 2-naphthalenesulfonates, nicotinates, napsllates, Nmethylglucamlnes, oxalates, octanoates, oleates, pamoates, fenllacetates, picrates, phenylbenzoates, pivalates, propionates, phthalates, pectinates, phenllproplonates, succinates, pyruvatates, pyruvatates, pyruvatates, pyruvates, succinates tartarates, theophylline acetates, ptoluenesulfonates (tosylates), trifluoroacetates, terephthalates, tannates, theoclates, trlhaloacetates, triethyl iodide, tricarboxylates, undecanoates and valerates.

In one embodiment, examples of inorganic salts of carboxylic acids or phenols can be selected from ammonium, metals

alkalis including lithium, sodium, potassium, cesium; alkaline earth metals including calcium, magnesium, aluminum; zinc, barium, hills, quaternary ammoniums.

In another embodiment, examples of organic salts of carboxylic acids or phenols can be selected from arginine, organic amines that include aliphatic organic amines, alicyclic organic amines, aromatic organic amines, benzathines, t-butylamines, bentamines, (/ V-benzylphenethylamine), dicyclohexylamines, dlmethylamines, diethanolamines, ethanolamines, ethylenediamines, hydramlines, imldazoles, lysines, methylamines, meglamines, A / -methylD-glucamines, N, / V'-dibenzylethylenediamines, nicotinamides, organic amines, urines, pyridines, picolies, piperazines, procaine, tris (hydroxymethyl) methylamines, triethylamines, triethanolamines, trimethylamines, tromethamines, and ureas.

In one embodiment, salts can be formed through conventional means, such as by reacting the free base or free acid form of the product with one or more equivalents of the appropriate acid or base in a solvent or medium in which the salt it is insoluble or in a solvent such as water, which is removed under vacuum or through freeze drying or exchanging the ions of an existing salt for another ion or suitable ion exchange resin.

In one embodiment, the methods of this invention make use of a pharmaceutically acceptable salt of the compounds of this invention. In one embodiment, the methods of this invention do

use of a pharmaceutically acceptable salt of the compounds formula IA, l-XII. In one embodiment, the methods of this invention make use of an amine salt of the compounds of the formula

IA, l-XII of this invention. In one modality, the methods of this

Invention make use of a phenol salt of the compounds of formula IA, l-XII of this invention.

In one embodiment, the methods of this invention make use of a free base, free acid, uncharged or non-complex compounds of formula IA, l-XII and / or its isomer, pharmaceutical, hydrate, polymorph, or combinations of the same.

In some embodiments of this invention, the compounds of this invention comprise three phenyl groups, which are held together through an amide bond. In one embodiment, the compounds of this invention are uncharged structures. In one embodiment, the compounds of this invention are free base structures. In one embodiment, the compounds of this invention are free acid structures. In one embodiment, the compounds of this invention are non-complex structures. In one embodiment, the compounds of this invention are non-ionized structures. In one embodiment, the compounds of this invention are pharmaceutically acceptable salts. In one embodiment, some compounds of this invention include hydrochloride salts (HCI).

In one embodiment, the methods of this invention make use of an Isomer of a compound of formula IA, l-XII. In one embodiment, the methods of this invention make use of a

ΙΜΡΙ Mixican institute Di LA FROFIIDAB INDUSTRIA!

compound of formula IA, l-XII. In one embodiment, the methods of this invention make use of a hydrate of a compound of formula IA, l-XII. In one embodiment, the methods of this invention make use of a polymorph of a compound of formula IA, l-XII. In one embodiment, the methods of this invention make use of a metabolite of a compound of formula IA, l-XII. In one embodiment, the methods of this invention make use of a composition comprising a compound of formula IA, l-XII, as described herein, or, in another embodiment, a combination of isomer, metabolite, pharmaceutical, hydrate, polymorph of a compound of formula IA, l-XII.

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

In one embodiment, the term "isomer" encompasses optical isomers of the compound. In one embodiment, the term "isomer" encompasses stereoisomers of the compound. The compounds of this invention possess an amide bond, which may be in its cis or trans isomerization. It should be understood that the present invention encompasses any optically active form, or stereolsomeric form, or mixtures thereof, and the use of these for any application is considered to be within the scope of this invention.

In another embodiment, this invention further includes hydrates of the compounds. In one embodiment, the term "hydrate" refers to

hemihydrate, monohydrate known in the art.

dihydrate, trihydrate or others, such as 'is'

Synthetic Procedures

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

For example, a compound of formula IA:

where R · ,, R2, R3 and R4, j and k are as previously described;

It can be prepared through a procedure that involves reacting:

IMPI

MEXICAN INSTITUTE OF THE OWN EDA 1 »INDUSTRIAL

Laugh

NH2 (1) together with '(2) to produce

diphenylamine (3) is reacted with,

in the presence of a

where if Rj, R2, R3 and R4 are independently OH, O-Alq-R5R6 or O-Alq-heterocycle, then R1 ', R2', R3'and R4'are a protected hydroxyl group, where the protecting group is removed to obtain the hydroxyl free or optionally followed by reacting with CI-Alq-heterocycle or CI-Alq-NR5R6 To produce a compound of formula IA:

(IA)

where if Ri, R2, R3 and R4 are independently OH, O-Alq-R5R6 or O-Alq-heterocycle, then RM, R2', Ra' and R4'are R ^ R2, R3 and R4, respectively.

As another example, a procedure for the preparation of a compound of Formula IA:

R (IA) where Ri, R2, R3 and R4 are as described above, includes reacting:

with

in the presence of a base to produce:

(4)

ΙΜΡΙ

MEXICAN INSTITUTE Dt THE INDUSTRIAL PROPERTY

where if R1t R2> R3 and R4 are independently OH, O-Alq-R5R6 or O-Alq-heterocycle, then Rf, R2', R3'and R4'are a protected hydroxyl group, wherein the protecting group is removed to obtain the free hydroxyl or optionally followed by reacting with CI-Alq-heterocycle or CI-Alq-NR5R6 To produce a compound of formula IA:

(IA) where if R1f R2, R3 and R4 are independently OH, O-Alq-R5R6 or O-Alq-heterocycle, then Rf, R2', R3'and R4'are R1: R2, R3 and R4, respectively.

In one example, Compound II is prepared according to Example 1, and Figure 5.

In another example, Compound III is prepared according to Example 1, and Figure 5.

In one more example, a compound of Formula IV:

HO (IV)

can be prepared by reacting:

in the presence of a base to produce:

followed by deprotection of the protecting groups to produce Compound ¡V:

where P and P 'are the same or different protecting groups. In one example, Compound IV is prepared according to Example 2, and Figure 6.

In another example, Compound V is prepared according to

IMPI

MEXICAN INSTITUTE «THE INDUSTRIAL PROPERTY

Example 1, and Figure 5. ---- ~

In a further example, Compound VI is prepared according to Example 3, and Figure 7.

In another example, Compound Vil is prepared according to Example 1, and Figure 5.

In another example, Compound VIII is prepared according to Example 4, and Figure 5.

In another example, Compound IX is prepared according to Example 5 and Figure 8.

In another example, the hydrochloride of Compound X is prepared according to Example 5 and Figure 8.

In another example, Compound XI is prepared according to Example 1, and Figure 5,

In another example, 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, carbobenzoxl, methoxyethoxymethyl (MEM) ether, 2 - (tr! M et i I s ί I i I) eto xim et i I (SEM) ether, metllthiomethyl (MTM) ether, fenllthiomell (PTM) ether, azidomethyl ether, cyanomethyl ether, 2,2-dec I or ro -1,1 - dif I u or oet i I ether, 2-chloroethyl ether, 2bromoethyl ether, tetrahydropyran, I (THP) ether, 1-ethoxyethyl (EE) ether, phenacyl ether, 4-bromophenacyl ether, cyclopropylmethyl ether, allyl ether, propargyl ether, sopropyl ether, cyclohexyl ether, t-butyl ether, benzyl ether, 2,6-dimethylbenzyl ether, 4-methoxybenzyl ether, o-nitrobenzyl ether, 2,664 dichlorobenzyl ether,

3,4-dichlorobenc

(dimethylamino) carbonylbenzyl ether, 4-m et i I su I fin ¡I be nci I ether, 4antrilmetil ether, 4-picolil ether, he ptaf I uo ro-pt o I i lo, tetraf I uoro - 4 - p ¡ r¡di I ether, trlmethylsilyl (TMS) ether, t-butyldimethylsilyl (TBDMS) ether, tbutiId¡fen¡IsiIiI (TBDPS) ether, aryl trisopropyIsi (TIPS) ether, aryl formate, aryl acetate, levulinate aryl, aryl pivaloate, aryl benzoate, aryl 9-fluorinecarboxylate, aryl methyl carbonate, 1-adamantyl carbonate, t-butyl carbonate, 4-methylsulfinylbenzyl carbonate, 2,4-dimethylpent-3-yl carbonate, 2,2,2-trichloroethyl aryl carbonate, aryl benzyl carbonate, aryl carbamate, dimethyl I f osf ini I ester ( Dmp-OAr), dim et ¡I fo sf inothioni I ester (Mpt-OAr), diphenylphosphinothionyl ester (Dpt-OAr), aryl methanesu Ifonate, aryl toluenesulfonate or aryl 2-formylbenzenesulfonate.

The methods of this invention comprise the use of compounds ΙΑ, l-XII, where the process for the preparation of the compounds of this invention comprises the reaction of a diphenylamine with a benzoyl chloride in the presence of a base. Suitable bases include, for example, pyridine, triethylamine, K2CO3, Cs2CO3, Na2CO3, methylamine, imidazole, benzimidazole, histidine, tributylamine, or any combination thereof. In one embodiment, the base is pyridine.

The methods of this invention comprise the use of compounds IA, l-XII, where the process for the preparation of the compounds of this invention comprises the deprotection of a protected hydroxyl. In another modality, the conditions of

IMPÍ iÑrriilrroMBOCano Ot LA rtOPlKJAB INDUSTRIAL

lack of protection depend on the protective group. In some embodiments, the deprotection step involves hydrogenation in the presence of Pd / C. In another embodiment, deprotection involves reaction with BBr3. In another embodiment, the deprotection step comprises reaction with an acid.

In other examples, Compounds IA, l-XII are prepared according to Figures 5-8 and Examples 1-5.

Pharmaceutical Compositions

In some embodiments, this invention provides methods of use, which comprise administering a composition comprising the disclosed compounds. As used herein, "pharmaceutical composition" means a "therapeutically effective amount" of the active ingredient, ie, the compound of this invention, along with a pharmaceutically acceptable carrier or diluent. A "therapeutically effective amount", as used herein, refers to that amount that provides a therapeutic effect for a given condition and regimen of administration.

As used herein, the term "administer" refers to bringing a subject into contact with a compound of the present invention. As used herein, administration can be accomplished in vitro, i.e., in a test tube, or in vivo, i.e., in cells or tissues of living organisms, eg, humans. In one embodiment, the present invention encompasses administration of the compounds of the present invention to a male subject.

This invention provides, in other embodiments, pharmaceuticals of the compounds described herein. The term "pharmaceutical product" refers to, in other embodiments, a composition suitable for pharmaceutical use (pharmaceutical composition), for example, as described herein.

The compounds of the invention can be administered alone or as an active ingredient in a formulation. Thus, the present invention also includes pharmaceutical compositions of the compounds of Formula I, containing, for example, one or more pharmaceutically acceptable carriers.

Numerous standard references are available that describe procedures for preparing various formulations suitable for administering the compounds according to the invention. Examples of potential formulations and preparations are contained in, for example, Handbook of Pharmaceutlcal Excipients, American Pharmaceutical Association (current edition); Pharmaceutical Dosage Forms: Tablets (Lieberman, Lachman, and Schwartz, editors) current edition, published by Marcel Dekker, Inc., as well as Remington's Pharmaceutical Sciences (Arthur Osol, editor), 1553-1593 (current edition).

The mode of administration and dosage forms are closely related to the therapeutic amounts of the compounds or compositions that are desirable and effective for the given treatment application.

Suitable dosage forms include, but are not limited to,

oral, rectal, sub-lingual, mucosa, nasal, ophthalmic, 'subcutaneous, intramuscular, intravenous, transdermal, spinal, intrathecal, intra-articular, intra-arterial administration, its trunk, bronchial, lymphatic, and Intra-uterine, and other dosage forms for systemic supply of the active ingredients. Formulations suitable for oral administration are preferred.

To prepare such dosage forms, the active ingredient can be mixed with a pharmaceutical carrier according to conventional pharmaceutical mixing techniques. The vehicle can have a wide variety of forms depending on the form of preparation desired for administration.

To prepare the compositions in an oral dosage form, any of the usual pharmaceutical media can be employed. Thus, for oral liquid preparations, such as, for example, suspensions, elixirs, and solutions, suitable carriers and additives include water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like. For oral solid 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. Due to their ease of administration, tablets and capsules represent the most advantageous form of oral dosage unit. If desired, the tablets can be sugar coated or enteric coated using standard techniques.

For parenteral formulations, the vehicle will usually comprise sterile water, although other ingredients may be included, for example, Ingredients that aid solubility or preservation. Injectable solutions can also be prepared, in which case appropriate stabilizing agents can be used.

In some applications, it may be advantageous to use the active agent in a "vectorized" form, such as through encapsulation of the active agent in a liposome or other encapsulation medium, or through attachment of the active agent, eg, to through covalent bonding, chelation, or associative coordination, in a suitable biomolecule, such as those selected from proteins, llpoproteins, glycoproteins, and poüsaccharides.

The methods of treatment of the present invention using formulations suitable for oral administration can be presented as discrete units such as capsules, small sacs, tablets, or troches, each containing a predetermined amount of the active ingredient such as a powder or granules. Optionally, a suspension can be used in an aqueous liquor or a non-aqueous liquid, such as a syrup, an elixir, an emulsion, or a potion.

A tablet can be made through compression or molding, or wet granulation, optionally with one or more accessory ingredients. Compressed tablets can be prepared Mexican Institute

DS THE MOPÍEDAD

BíDumiAL

compressing in a suitable machine, with the active compound being in a free-flowing form such as a powder or granules, which optionally can be mixed with, for example, a binder, disintegrating agent, lubricant, inert diluent, active agent in the surface, or discharge agent. Molded tablets composed of a mixture of the powdered active compound with a suitable vehicle can be made by molding on a suitable machine.

A syrup can be made by adding the active compound to a concentrated aqueous solution of a sugar, for example sucrose, to which any accessory ingredient (s) can also be added. Said accessory ingredient (s) includes flavorings, suitable preservatives, agents to delay the crystallization of sugar, and agents to increase the solubility of any ingredient, such as an alcohol ρ or I ih dro χ ί I co, for example, glycerol or sorbitol.

Formulations suitable for parenteral administration may comprise a sterile aqueous preparation of the active compound, which is preferably isotonic with the blood of the recipient (eg, physiological saline). Such formulations can include suspending agents and thickening agents and liposomes or other microparticle systems, which are designed to target the compound to components of the blood or one or more organs. The formulations can be presented in either a single dose or multiple dose form.

Mexican Institute pt la TRorir.oM)

INDUSTRIAL

Parenteral administration can comprise any suitable form of systemic delivery. Administration, for example, can be intravenous, intra-arterial to I, intrathecal, intramuscular, subcutaneous, intramuscular, intra-abdominal (for example, intraperitoneal), etc., and can be carried out by infusion pumps (external or can be implanted) or any other means appropriate to the desired mode of administration.

Nasal formulations and other sprays through the mucosa (eg, inhalable forms) may comprise purified aqueous solutions of the active compounds with preservatives and isotonic agents. Said formulations preferably adjust to a pH and isotonic state compatible with the nasal membranes or other mucosa membranes.

Alternatively, they can be in the form of finely divided solid powders, suspended in a gaseous vehicle. Such formulations can be delivered through any suitable means or method, for example, through a nebulizer, atomizer, metered dose inhaler, or the like.

The rectal administration formulations can be presented as a suppository with a suitable vehicle such as cocoa butter, hydrogenated fats, or hydrogenated fat carboxylic acids.

Transdermal formulations can be prepared by incorporating the active agent in a txxotropic or gelatinous vehicle such as a cellulosic medium, eg, methylcellulose or

IMPI

MEXICAN INSTITUTE • E industrial PROPERTY

hydroxyethyl cellulose, the resulting formulation is then wrapped in a transdermal device adapted to be secured in dermal contact with a user's skin.

In addition to the aforementioned ingredients, the formulations of this invention may further include one or more accessory ingredients selected from, for example, diluents, pH regulators, flavoring agents, binders, disintegrating agents, surface active agents, thickeners, lubricants. , preservatives (including antioxidants), and the like.

The formulations of the present invention may have immediate release, sustained release, delayed onset release or any other release profile known to one skilled in the art.

In one embodiment, this invention provides methods for a) reducing serum total testosterone levels; b) reducing serum free testosterone levels through the decrease of luteinizing hormone (LH) or independent of the decrease of LH hormone in a male subject with prostate cancer, which comprises administering an oral composition comprising a compound of formula IA, l-XII. In further embodiments, the methods of this invention make use of an oral composition comprising a compound of formula II, formula lll, formula IV, formula V, formula VI, formula Vil, formula VIII, formula IX, formula X, formula XI or formula XII

In one embodiment, this invention provides a method for

ΙΜΡΙ

MEXICAN INSTITUTE OE LA PROHEDAD INDUSTRIAL

the treatment of prostate cancer by decreasing 105 n'IVSlSS from W or independent of the decrease in LH levels in a male subject having prostate cancer, comprising administering an oral composition comprising a compound of formula IA, l-XII . In further embodiments, this invention provides methods for treating prostate cancer by lowering LH levels or Independent of lowering LH levels in a male subject having prostate cancer, which comprises administering an oral composition comprising a compound of formula II, formula III, formula IV, formula V, formula VI, formula Vil, formula VIII, formula IX, formula X, formula XI or formula XII.

It should be understood that this invention encompasses any embodiment of a compound as described herein, which in some embodiments is referred to as a "compound of this invention".

In one embodiment, the methods of this invention may comprise administering a compound of this invention at various doses. In one embodiment, a compound of this invention is administered at a dose of 1-1500 mg per day. In further embodiments, a compound of this invention is administered at a dose of 1-10 mg per day, 3-26 mg per day, 3-60 mg per day, 3-16 mg per day, 3-30 mg per day, 10-26 mg per day, 15-60 mg, 50-100 mg per day, 50-200 mg per day, 1 50-300 mg per day, 20-50 mg per day, 5-50 mg per day, 200 -500 mg per day, 1 50-500 mg per day, 200-1000 mg per day, 300-1 500 mg per day or 1 00-1000 mg per day.

ΜΡϊ

Μ.Τ, Τ ', τ ,, MEX,SPOUT '· Λ -OHÍDAD ^ P' ^ TMIAL

In one embodiment, the methods of this invention may comprise administering a compound of this invention at various doses. In one embodiment, a compound of this invention is administered at a dose of 0.1 mg / kg / day. In further embodiments, a compound of this invention is administered at a dose of between 0.2 to 30 mg / kg / day, or 0.2 mg / kg / day, 0.3 mg / kg / day, 1 mg / kg / day, 3 mg / kg / day, 5 mg / kg / day, 10 mg / kg / day, 20 mg / kg / day or 30 mg / kg / day.

In one embodiment, the methods of this invention are provided to use a pharmaceutical composition comprising a compound of the formula ΙΑ, 1-12. In further embodiments, the methods of this invention are provided to use a pharmaceutical composition comprising a compound of formula ll, formula III, formula IV, formula V, formula VI, formula Vil, formula VIII, formula IX, formula X, formula XI or formula XII.

In certain embodiments, the pharmaceutical composition is a solid dosage form. In another embodiment, the pharmaceutical composition is a tablet. In another modality, the composition

MEXICAN INSTITUTE

OF IA PROPERTY

INDUSTRIAL

pharmaceutical is a capsule. In another embodiment, the pharmaceutical composition is a solution. In another embodiment, the pharmaceutical composition is a transdermal patch.

--- In -me modality, e) use of eum-c om position of this -invention o_______ a composition comprising the same, will have utility to inhibit, suppress, improve or stimulate a desired response in a subject, as will be understood by one skilled in the art. In another embodiment, the compositions may further comprise additional active ingredients, the activity of which is useful for the particular application for which the compound of this invention is being administered.

For administration to mammals, and particularly humans, the physician is expected to determine the actual dose and duration of treatment, which is most suitable for an individual and may vary with the age, weight, genetics, and / or response of the particular individual. .

In some embodiments, any of the compositions of this invention will comprise a compound of this invention, in any form or embodiment described herein. In some embodiments, any of the compositions of this invention will consist of a compound of this invention, in any form or embodiment described herein. In some embodiments, the compositions of this invention will consist essentially of a compound of this invention, in any form or embodiment described herein. In some modalities, the term "includes" refers to the inclusion of the

MEXICAN INSTITUTE '?

OE LA MOHEDA »

INDUSTRIAL

indicated active agent, such as the compound of this invention, as well as the inclusion of other active agents, and pharmaceutically acceptable carriers, excipients, emollients, stabilizers, etc., as is known in the pharmaceutical industry. In some embodiments, the term "consisting essentially of" refers to a composition, the sole active ingredient of which is the indicated active ingredient, however, other compounds may be included, which are to stabilize, preserve, etc., the formulation , but are not directly involved in the therapeutic effect of the indicated active ingredient. In some embodiments, the term "consisting essentially of" may refer to components that facilitate the release of the active ingredient. In some embodiments, the term "consisting of" refers to a composition, which contains the active ingredient and a pharmaceutically acceptable carrier or excipient.

It should be understood that any use of any of the compounds described herein can be used in the treatment of any disease, disorder or condition as described herein, and represents an embodiment of this invention. In one embodiment, compounds with a base compound free, acid free, uncharged, or not in complex.

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

EXAMPLES

EXAMPLE 1

General synthesis procedures for compounds of Formulas ll-XII and synthetic intermediates

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

The estrogen receptor ligands described herein can be prepared in a number of ways well known to those skilled in the art. For example, the estrogen receptor ligands described herein can be prepared through synthetic methods described in the Patent Application Publication of

USA No. 2009/0062341, the descriptions of which are incorporated herein by reference in their entirety.

General Synthesis of Derivatives of Λί, ΛΓ-bis Aryl Benzamide

General synthesis of diarylanilines (Figure 5). A mixture of arilamlna (1.5 equivalents), aryl iodide (1 equivalent), K2CO3 (2 equivalents), Cul (0.1 equivalents) and L-proüna (0.2

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INDUSTRIAL equivalents) was mixed together and dissolved in anhydrous DTVTSO '' ”at room temperature. Then, the reaction mixture was stirred and heated at 90 ° C for 28 hours. The mixture was cooled to room temperature and hydrolyzed with water. EtOAc was added to split the solution. The EtOAc layer was separated, washed with brine, and dried over MgSO4. The solvent was removed under reduced pressure. The solid residue was purified by flash column chromatography (silica gel) using 5% EtOAc / hexanes as the eluent to provide the corresponding dialaniline.

Bis- (4-methoxyphenyl) amine (1a): pale yellow solid, 73% yield. Mp 98.6-99.0 ° C.1H NMR (CDCÍ3, 300 MHz) δ 6.936.81 (m, 8H), 5.37 (s, br, 1H), 3.78 (s, 6H). MS m / z 228.4 (MH)+

N- (4-Methoxyphene yl) -phenylamine (Ib): pale yellow solid, 70% yield. Mp 106.3 -1 06.5 ° C.1H NMR (CDCI3, 300 MHz) δ 7,247.18 (m, 3H), 7.08-7.06 (m, 2H), 6.92-6.84 (m, 4H), 5.61 (s, br, 1H), 3.79 (s, 3H). MS m / z 200.1 (M + H)+ .

N- (4-Fluorophenyl) -N-4-methoxyphenylamine (le): pale yellow solid, 54% yield. Mp 60.6-6t.0 ° C.1H NMR (CDCI3, 300 MHz) δ 7.01-6.83 (m, 8H), 3.78 (s, 3H). MS m / z 217 (M)+ .

N- (4-Benzyloxyphenyl) -N-4-methoxyphenylamine (Id): pale yellow solid, 54% yield. Mp 1 08.0-108.4 ° C.1H NMR (CDCI3, 300 MHz) δ 7.34-7.08 (m, 5H), 6.90-6.81 (s, 3H), 3.78 (s, 3H). MS m / z 306 (M + H)+ .

General synthesis of Benzamides. A mixture of arylaniline (1 equivalent), benzoyl chloride (1.3 equivalents), and pyridine (6 equivalents) 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 through the addition of a 2N HCI solution. The solution was extracted with ethyl acetate. The organic layer was washed with a NaHCO solution3 saturated aqueous to remove excess acid, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified through flash column chromatography using EtOAc / hexanes (3/7 v / v) to give the corresponding benzamide compounds.

3- Fluoro-N- (4-fluorophenyl) -4-methoxy-N- (4-methoxyphenyl) benzamide (2a): yellow solid, mp 54-56 ° C, 1H NMR (CDCI3/ TMS) 57.24-7.1 1 (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]+

4- Fluoro-N, N-bis (4-methoxyphenyl) -2- (trfl uoromethyl) benzamide (2b): Colorless oil, 84.2% yield. 1H NMR (CDCI3, 300 MHz) δ 7.34-7.26 (m, 4H), 7.09-7.01 (m, 3H), 6.91 (dv 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)+ .

4-Wletox¡-N- (4-methoxiphenyl) -N- (4-fluorophenyl) -benzamida (2c):

white solid, 97% yield, mp 1 33.5.0-134.5 ° C. 1H NMR (CDCI3, 300 MHz) δ 8.11-6.66 (m, 15H), 3.74 (s, 3H), 3.73 (s, 3H).

MS m / z 384 (M + H) + .

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N- (4-Methoxyphenyl) -N- (4-benzyloxyphenyl) -2-naphthylamide f2d): - white solid, 58% yield. Mp 174.9-1 75.5 ° C.1H NMR (CDCI3, 300 MHz) δ 8.04 (s, 1H), 7.77-7.74 (m, 2H), 7.64-7.61 (m, 1H), 7.517.43 (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)+ .

4-Fluoro-N, N-bis (4-methoxy fe nil) -2- (trifluoromethyl) be nzamide (2e): Colorless oil, 84.2% yield. 1H NMR (CDCI3, 300 MHz) δ 7.34-7.26 (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)+ .

General Procedure for Demethylation of Benzamide Derivatives Using BBr3. A methoxybenzamide compound was dissolved in CH2CI2 dry. BBr added3 (1.0 M CH solution2CI2) drop by drop at 0 ° C. The reaction solution was slowly warmed to room temperature and allowed to stir overnight at room temperature. The mixture was cooled to 0 ° C in an ice bath and hydrolyzed by adding water. EtOAc was added to split the solution. The organic layer was separated; the aqueous layer was extracted with EtOAc. The organic layer was washed with brine and dried over MgSO4 anhydrous. The solvent was removed under reduced pressure. The residue was purified through flash column chromatography using CH3OH / CH2CI2 (1/9 v / v) to give the corresponding phenolic compounds.

4-Fluoro-N, Wb¡s (4-hydroxyphenyl) -2- {trifluoromethyl) benzamida (3a):

white solid, 92.5% yield. 1H NMR (DMSO-J8, 300 MHz) δ

9.55 (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)+ .

The following compounds were synthesized as described hereinabove and are characterized and summarized in Table 1: N, / \ - - bis (4-hydroxyphenyl) -4-propylbenzamide (II); 3-fluoro- / V (4-fluorophenyl) -4-hydroxy- / V- (4-hydroxyphenyl) benzamide (IV); W, A / -bis (4hrodroxfenll) -2,3-dimethylbenzamide (V); 3-fluoro-4-hldroxy- / V, A / -bis (4hydroxyphenyl) -benzamide (Vil); 3-fluoro-4- hydroxy-N- (4-hydroxyphenyl) -A / phenylbenzamld (XI); and 3-f I or gold- / V, ΛΖ-bis (4-h idrox if in i I) -2methylbenzamide (XII).

General Procedures for the Debenzylation of Benzyloxyphenyl-benzamides. The compound was dissolved in EtOH in a 250 ml hydrogenation bottle. Pd / C powder (5 mol%) was added to the solution. The reaction vessel was mounted in a hydrogenation apparatus under a pressure of 1,406 kg / cm2 hydrogen gas. The reaction was verified through TLC until the starting material disappeared. Then, the solvent was removed under reduced pressure. The residue was purified through flash column chromatography with hexanes / EtOAc = 3/2 v / v to give the desired product.

The following compounds were synthesized as previously described herein and are characterized and summarized in

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Table 1: N, N-bs (4-hydroxyphenyl, I) -2-naphthylamide (IV).

General Procedures for the Reduction of Unprotected Benzamides. Benzamide compounds were dissolved in 20 ml of anhydrous THF at room temperature. H added3B (SMe2) through a syringe at room temperature under argon. The reaction solution was stirred and heated under reflux for 6 hours. The reaction was then quenched by adding 10 ml of MeOH at 0 ° C. The solvent was removed under reduced pressure. The residue was subjected to flash column chromatography (silica gel, CH2CI2/ MeOH = 9/1 v / v) to give the desired product.

The following compounds were synthesized as described hereinabove and are characterized and summarized in Table 1: 4,4 '- (2,3-dimethi I benzyl I azanodii I) d if e η oi (III); 4 - ((4fluorophenyl) (4-hydroxybenzyl) amino) phenol (VIII).

General synthesis of 0- (2-piperidin-1-ylethoxy) -benzamides and the like. To a solution of the hydroxyphenyl-containing benzamide analog (1 equivalent) in acetone, K was added2CO3 (3 equivalents) and N-chloroethyl-piperidine hydrochloride salt (1.2 equivalents). The solution was heated under reflux for 6 hours. The solution was evaporated to dryness. The residue was hydrolyzed by adding water, and then extracted with ethyl acetate. The organic layers were separated and dried over MgSO4 anhydrous. The solvent was removed under reduced pressure. The residue was purified through

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flash chromatography with methylene chloride / methanol = 9/1 v / v to give the desired compound '. Hydrochloride salts were prepared by adding HCI in Et2Or to the methanol solution of the compounds followed by the evaporation of solvents.

The following compounds were synthesized as previously described herein and are characterized and summarized in Table 1: 4-fluoro-N- (4-hydroxyphenyl) -N- (4- (2- (p¡per¡d¡n -1-yl) ethoxy!) Phenyl) -2- (trifluoromethyl) benzamide (IX); and 4-fluoro- / V- (4hid rox if eni I) -Λ / - (4- (2 - (pi per id i n-1 - i I) eto xi) feni I) - 2 - (t rif I uo rom et i I) benzamide (X) which is the HCI salt of IX.

TABLE 1. Physical Characterization of the Compounds of the

Formulas ll-XII

<td>Comp. #</td><td>Structure</td><td>PHYSICAL CHARACTERIZATION</td>

<td>II.</td><td>Oh</td><td>1H NMR (DMSO-dg, 300 MHz) 69.46 (s, 2H, 2 X OH), 7.27-7.26 (m, 2H, ArH), 7.06-7.04 (m, 2H, ' ArH), 6.99-6.97 (m, 4H, ArH), 6.66-6.65 (m, 4H, ArH), 2.50 (s, 2H, CH2, overlapped with DMSO peak), 1.53-1.52 (m, 2H, CH2), 0.82 (t, J = 7.33 Hz, 3H, CH3). m / z 346.0 (MH) '</td>

<td>III.</td><td>Η° Ά jCu Φ Oh</td><td>Cinnamon foam, 41% rend. Mp 147-150 ° C.1H NMR (DMSO-de, 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), 6.63-6.59 (m, 4H), 4.72 (s, 2H), 2.23 (s, 3H), 2.16 (s, 3H). m / z 320.2 (M + Hf</td>

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<td> 5</td><td>IV.</td><td>φΤΧ F</td><td>Solid cinnamon, 92% rend. Mp 110-112 ° C.1H NMR (DMSO-de, 300 MHz) δ 10.14 (bs, 1H). 9.71 (bs, 1H) 7 26-711 (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)+</td>

<td> 10</td><td>V.</td><td>ocHliyCH:' Oh</td><td>1H NMR (DMSO-d6, 300 MHz) δ 9.47 (bs, 2H, 2 X OH), 7.18 (d, J = 8.30 Hz, 2H, ArH), 7.06 (d, J = 7.08 Hz, 1H, ArH), 7.00-6.92 (m, 4H, ArH), 6.78 (d, J = 8.30 Hz, 2H, ArH), 6.51 (d, J = 8.06 Hz, 2H, ArH), 2.22 (s, 3H, CH3), 2.15 (s, 3H, CH3). m / z 334.3 (M + H)+</td>

<td> 15</td><td>SAW.</td><td>Oh</td><td>White solid, 70% rend. Mp 264.3-265.2DC (decomposed). 1H NMR (DMSO-d6, 500 MHz) δ 9.46 (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)+</td>

<td> 20</td><td>HIV.</td><td>Χχχχ F</td><td>Yellow oil, 92% rend. 1H NMR (DMSO-d6, 500 MHz) (59.29 (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) '</td>

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<td>XI yX. (Get ouf of HCI of IX.)</td><td>τρ-χι. ϊΓ · Oh</td><td>White solid, 57.7% yield. 1H NMR (DMSO-d6, 300 MHz) δ 9.57 (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, J = 5.7 Hz), 2.55 (t, 1H, J = 5.4 Hz), 2.44 (s, 211), 2.36 (s, 2H), 1.49-1.37 (m, 6H). m / z 501.0 (MH) '</td>

<td>XII.</td><td>ü i S fHsOh</td><td>Pale red solid. 72.0% yield Mp> 240 ° C.1H NMR (DMSO-de, 300 MHz) δ 9.50 (bs, 2H), 7.19- 6.79 (m, 7H), 6.61 (d, J = 8.93 Hz, 2H), 6.53 (d, J = 7.79 Hz, 2H), 2.23 (s, 3H). m / z 336.0 (MH) '</td>

EXAMPLE 2

Synthesis of the compound of formula IV (Figure 6)

Cul, L-proline, K2CO3 'OCH3 at DMSO, 90-95 ° C, o / n

Step 1:

A

Synthesis of 4-fluoro- / V- (4-methoxyphenyl) aniline (1c) 4-fluoroaniline mixture (78.63 g, 0.708 mol), 4yodoanisole (1 38.00 g, 0.590 mol), K2CO3 anhydrous (1 22.23 g, 0.884

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moles), Cul (11.23 g, 58.96 mmol) and L-proline (13.58 g, 0.118 moles) were mixed together in a 1 liter 3 neck round bottom flask equipped with a stir bar, a reflux condenser and a argon inlet. Anhydrous DMSO (300 ml) was added at room temperature. The reaction mixture was stirred and heated at 90 ° C for 20 hours under argon. The mixture was then cooled to room temperature and hydrolyzed with water (300 ml). EtOAc (200 ml) was added to divide the solution. The EtOAc layer was separated. The aqueous layer was extracted with 100 ml of EtOAc. The EtOAc layers were combined, washed with brine (2x100 ml), and dried over MgSO4 anhydrous (50 g). The solvent was removed under reduced pressure. The brown oily residue was purified by flash chromatography (silica gel, hexanes / EtOAc = 9/1 v / v) to give 4-fluoro- / V- (4methox¡fen¡l) anlna (1c) as a yellow solid product, 99.70 g, 77.8% yield. Mp 46-48 ° C. MS (ESI) m / z 218.1 [M + H]+, 1H NMR (DMSO-J6, 300 MHz) δ 7.77 (bs, 1H), 7.03 - 6.98 (m, 4H), 6.93 - 6.82 (m, 4H), 3.70 (s, 3H).

you

2nd

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Step 2: Synthesis of 3-fluoro-N- (4-fluorophenyl) -4-methoxy-W- (4methoxyphenyl) benzamide (2a)

4-Fluoro- / V- (4-methoxyphenyl) aniline (1c) (90.78 g, 0.418 mol) and 3-fluoro-4-methoxybenzoyl chloride (94.55 g, 0.501 mol) were mixed together and dissolved in THF anhydrous (200ml) in a 1 liter three neck round bottom flask equipped with a stir bar, a reflux condenser and an argon inlet. Anhydrous pyridine (1,32.22 g, 1,672 mol) was added via syringe at room temperature under argon - the reaction mixture was stirred and refluxed overnight. The reaction mixture was then cooled to room temperature and filtered to remove the pyridine salt. The solution was concentrated to remove the THF solvent. The residual oil was washed with 200 ml of a 2N HCI solution and extracted with ethyl acetate (2 x 200 ml). The organic layer was washed with a Na solution2CO3 (150 ml) aqueous saturated to remove excess benzoyl chloride and acid, dried over MgSO4 (50 g), filtered, and concentrated under reduced pressure to give an oil. The residue was purified through flash column chromatography using silica gel with CH2CI2/ acetone (50/1 v / v) to provide the corresponding pure benzamide compound as a yellow solid. Mp 54-56 ° C. MS (ESI) m / z 370.1 [M + H]+ , 1H NMR (CDCI3/ TMS) or 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).

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Step 3: Synthesis of 3-fluoro-N- (4-fluorophenyl) -4-h¡droxi- / V- (4h¡droxifen¡l) benzamida (IV)

The 3-fluoro-N- (4-fluorophenyl) -4-methoxy-A / - (4methoxyphenyl) benzamide (2a) compound (138.0 g, 0.374 mol) was dissolved in CH2 Cl2 dry (600 ml) at room temperature under argon. BBr added drop by drop3 (374.75 g, 1,496 moles) with stirring through a syringe at 0 ° C in an ice bath under argon. The reaction solution was allowed to stir at room temperature overnight. The solution was then emptied into 1 liter of ice water with 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. CH layer2CI2 separated, dried over MgSO4 anhydrous (50 g), filtered and concentrated under reduced pressure to dryness. The white precipitate and the residue of the CH solution2CI2 pooled and purified via flash column chromatography (silica gel, CH2CI2/ Acetone / MeOH = 90/7/3 v / v / v) to give a tan solid, which was recrystallized from hot EtOAc / hexanes twice to provide a white crystalline solid, 104.0 g, 81.6% yield. Mp 110-112 ° C. MS (ESI) m / z 364.1 [M + Naf,1H NMR

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INDUSTRIAL (DMSO-J6) δ 10.14 (bs, 1H), 9.71 (bs, 1H), 7.25-7.11 (m, 5H), 7,056.99 (m, 3H), 6.78 (t, J = 8.6 Hz, 1H), 6.68 (d, J = 8.7 Hz, 2H).

EXAMPLE 3

Synthesis of the compound of formula VI (Figure 7)

Synthesis of 4- (benzyloxy) -W- (4-methoxyphenyl) aniline (Id)

A mixture of 4-benzyloxyaniline (16.6 g, 83.31 mmol), 4-iodoanisole (15.0 g, 64.09 mmol), K2CO3 (17.72g, 128.18mmol), Cul (1.22g, 6.41mmol) and L-proline (1.48g, 12.82mmol) were combined together and dissolved in anhydrous DMSO (120ml) at room temperature. Then, the reaction mixture was stirred and heated at 90 ° C for 48 hours. The mixture was cooled to room temperature and hydrolyzed with water. EtOAc was added to split the solution. The EtOAc layer was separated with brine, dried over MgSO4 anhydrous. The solvent was removed under reduced pressure. The solid residue was purified through flash column chromatography (silica gel) using EtOAc / hexanes (1/9 v / v) to provide the corresponding d i a ri I an I ina as a yellow solid , 9.8 g, 50% yield. Mp 1 08.0-1 08.4 ° C. 'H NMR (CDCIs, 300 MHz) δ 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)+ .

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Synthesis of Af- (4-benzyloxyphenyl) -N- (4-methoxyphenyl) -2-nafthamide (2d)

An equivalent of 4- (benzyloxy) - / \ / - (4-methoxyphenyl) aniline (0.80 g, 2.62 mmol) was mixed with 1.5 equivalents of 2-naphthoyl chloride (0.75 g, 3.93 mmol) and 4 pyridine equivalents (0.83 g, 10.48 mmol) in a dry three-necked round bottom flask equipped with a magnetic stir bar and a reflux condenser. The mixture was dissolved in anhydrous THF (30 ml) and heated under reflux for 20 hours. The reaction solution was cooled to room temperature and filtered. The solvent was removed under reduced pressure. The residue was purified through flash column chromatography using silica gel with EtOAc / hexanes (3/7 v / v) to provide the corresponding pure naphtamide compound as a white solid, 0.70 g, 58% yield . Mp 1 74.9-175.5 ° C.1H NMR (CDCI3, 300 MHz) δ 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)+ .

Synthesis of / V, N-pis (4-hydroxyphenyl) -2-naphthylamides (VI)

Compound A / - (4-benzyloxyphenyl) - / V- (4-methoxyphenyl) -2nafthamide (2d) (0.50 g, 1.09 mmol) was dissolved in CH2 Cl2 (30 ml) dry at room temperature. BBr added drop by drop3 (3.26 ml of a 1.0 M CH solution2CI2, 3.26 mmol) with stirring through a syringe at room temperature. The reaction solution is .ertlA .- 'tWJi'V.

ΙΜΡΪίINSTmiTO mexicana i and Di LA ΜΟΜΕΒλΡ> INDUSTRIAL allowed to stir overnight at room temperature. The mixture was cooled to 0 ° C in an ice bath and hydrolyzed by adding water. EtOAc was added to split the solution. The organic layer was separated; the aqueous layer was extracted with EtOAc twice. The organic layers were combined, washed with brine, and dried over MgSO4 anhydrous. The solvent was removed under vacuum. The residue was purified through flash column chromatography using silica gel with CH3OH / CH2CI2 (1/9 v / v) to provide the desired pure phenolic compound as a white solid, 0.27 g, white solid, 70% yield. Mp 264.3-265.2 ° C (decomposed). NMR (DMSO-J6, 500 MHz) δ 9.46 (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 + Hf.

EXAMPLE 4

Synthesis of the compound of formula VIII

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

The compound / V- (4-fluorophenyl) -4-hydroxy- / V- (hydroxyphenyl) benzamlda (0.30 g, 0.93 mmol) was dissolved in 20 ml of anhydrous THF at room temperature . H added3B (SMe2) (1.86 ml of

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INDUSTRIAL ______ a solution of 2M THF, 3.71 mmol) through a syringe at room temperature under argon. The reaction solution was stirred and heated under reflux for 6 hours. The reaction was then quenched by adding 10 ml of MeOH at 0 ° C. The solvent was removed under reduced pressure. The residue was subjected to flash column chromatography (silica gel, CH2CI2/ MeOH = 9/1 v / v) to give a yellow oil. 0.26 g, 92% yield.1H NMR (DMSO-J6, 500 MHz) δ 9.29 (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 the compound of formulas IX and X (Figure 8)

Synthesis of diarylanilines. A mixture of arilamlna (1.5 equivalents), aryl iodide (1 equivalent), K2CO3 (2 equivalents), Cul (0.1 equivalent), and L-proline (0.2 equivalent) were combined together and dissolved in anhydrous DMSO at room temperature. Then, the reaction mixture was stirred and heated at 90 ° C for 28 hours. The mixture was cooled to room temperature and hydrolyzed with water. EtOAc was added to split the solution. The EtOAc layer was separated, washed with brine, dried over MgSo4 anhydrous. The solvent was removed under reduced pressure. The solid residue was purified through column chromatography of

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instant vaporization (silica gel) using EtOAc / hexanes (3/7 v / v) as solvent to provide the corresponding diarylaniline. Bis- (4-methoxyphenyl) amine (la): pale yellow solid, 73% yield.1H NMR (CDCI3, 300 MHz) δ 6.93-6.81 (m,

8H), 5.37 (s, br, 1H), 3.78 (s, 6H). MS m / z 228.4 (MH)+ .

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

1 equivalent of bis- (4-methoxyphenyl) amine (la) (0.73 g, 3.18 mmol) was mixed with 1.2 equivalents of 4-fluoro-2trifluoromethylbenzoiium chloride (0.87 g, 3.82 mmol) and 6 equivalents of pyridine (1.51 g, 19.08 mmol) in a dry three-necked round-bottom flask equipped with a magnetic stir bar and a reflux condenser. The mixture was dissolved in anhydrous THF (20 ml) and heated at 90 ° C for 20 hours. The reaction solution was cooled to room temperature and filtered. The solvent was removed under reduced pressure. The residue was purified through flash column chromatography using silica gel with EtOAc / hexanes (3/7 v / v) to provide the corresponding pure benzamide compound as a colorless oil, 1.12 g, 84.2% yield.1H NMR (CDCI3, 300 MHz) δ 7.34-7.26 (m, 4H), 7.097.01 (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)+ .

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Synthesis of 4-flu gold- W, / V-bis (4-hydroxyphenyl) -2 - '(trif luoromethyl) benzamide (3a)

Compound 4-fluoro-N, N-bis (4-methoxyphenyl) -2- (trifluoromethyl) benzamide (2e) (1.00 g, 2.38 mmol) was dissolved in CH2CI2 dry (30 ml) at room temperature. BBr added drop by drop3 (10 ml of a 1.0 M CH solution2CI2, 10.0 mmol) with stirring through a syringe at room temperature. The reaction solution was allowed to stir overnight at room temperature. The mixture was cooled to 0 ° C in an ice bath and hydrolyzed by adding water. EtOAc was added to split the solution. The organic layer was separated; the aqueous layer was extracted with EtOAc twice. The organic layers were combined, washed with brine, and dried over MgSO4 anhydrous. The solvent was removed under vacuum. The residue was purified by flash column chromatography using silica gel with ΟΗ3ΟΗ / ΟΗ2ΟΙ2 (1/9 v / v) to provide the desired pure phenolic compound as a white solid, 0.86 g, 92.5% yield. 1H NMR (DMSO-J6, 300 MHz) δ 9.55 (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)+ .

Synthesis of 4-fluoro-N- (4-hydroxyphenyl) -N- [4- (2-piper¡din-1-yl) ethoxyphene l] -2- (trifl uorometi Ijbenzamida. (IX)

To a solution of 4-fluoro- / V, A / -bis (4-hydroxyphenyl) -2 (trifluoromethyl) benzamide (3a) (0.61 g, 1.56 mmol) in acetone,

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY added K2CO3 (1.29 g, 9.36 mmol) and N-chloroethyl-piperidine hydrochloride salt (0.34 g, 1.87 mmol). The solution was heated under reflux for 20 hours. The solution was evaporated to dryness. The residue was purified through flash chromatography (silica gel; methylene chloride / methanol = 9/1 v / v) to give the desired compound as a white solid, 0.45 g, 57.7% yield.1H NMR (DMSO-J6, 300 MHz) δ 9.57 (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, 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) '.

The hydrochloride salt (X) was prepared by adding HCI in Et2Or to the methanol solution of the compounds, followed by evaporation of the solvents.

EXAMPLE 6

Estrogen receptor binding affinities, Agonist and Antagonist activity

The ER binding affinity of the compounds was determined using a competitive in vitro radlollgando binding assay with [2,4,6,7 -3 Η (N) j - E st ra dio I ([3H] E2), a naturally occurring high affinity ER ligand, and a bacterially expressed GST fusion ER-α or ER-β ligand binding domain (LBD) protein.

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Method

Recombinant ER-α or ER-β was combined with [3H] E2 to determine the equilibrium dissociation constant (Kd) of [3H] E2. The protein was incubated with increasing concentrations of [3H] E2 with and without a high concentration of unlabelled E2 at 4 ° C for 18 hours in order to determine total and non-specific binding. Non-specific binding was subtracted and the Kd of E2 (ERa: 0.71 nM; ΕΡβ: 1.13 nM) was determined using nonlinear regression. Furthermore, the concentration of [3H] E2 was required to saturate ER-α and ER-β was determined to be 4-6 nM.

Increasing concentrations of compounds (scale: 10 '11 to 10 5 M) were incubated with [3H] E2 (5.7 nM) and ER LBD using the conditions described above. After incubation, plates were harvested with GF / B filters on the Unifilter96 Harvester harvester (Perkin Elmer) and washed three times with ice cold pH B regulator (50mM Tris, pH 7.2). The filter plates were dried at room temperature, then 35 µΙ of the Mrosccntnt-0 cocktail was added to each well and the filter plates were sealed with TopSeal-A. Radioactivity was counted on a TopCount® NXT microplate clipping counter using the parameters to 3H in the Microscint cocktail (Perkin Elmer).

The specific union of [3H] E2 at each concentration of the compounds it was determined by subtracting the nonspecific binding of [3H] E2 (determined by incubating with 10 '6 M of unlabeled E2) and expressing it as a percentage of the specific binding in the absence

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MEXICAN INSTITUTE DF. THE INDUSTRIAL PROPERTY of the test compound. We determined the concent'racToñ 3e ~ TüT compounds that reduced the specific binding of [3H] E2 at 50% (IC50). The equilibrium union constant (K¡) of the compounds was then calculated through: K¡ = Ktí χ Ι05ο / (Κό + L), where Κύ is the equilibrium dissociation constant of [3H] E2 (ER-α = 0.71 nM; ER-β = 1.13 nIVI), and L is the concentration of [3H] E2 (ER-a: 5.7 nM; ER-β: 5.7 nM).

Results

Binding assays revealed that ER-α and ER-β bound ligands at various concentrations, ranging from 3.75 nM to over 1000 nM and the selectivity varies from the compound being selective for isoform to being selective for non-isoform. Representative compound results are listed in Table 2.

TABLE 2. Binding Results for Selected Compounds

<td>COMPOUNDS</td><td>ER-α Ki (nM)</td><td>ER-β Ki (nM)</td>

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AND! compound IV binds ERa and ERp. The ER binding affinity of Compound IV was determined using a competitive in vitro radioligand binding assay with | 2,4,6,7-3H (N)] - Estradiol ([3H] E2), a naturally occurring high affinity ER ligand, and a GST fusion ER-α or ER-β ligand binding domain (LBD) protein

bacterially expressed. In this test, the binding affinities of ERa and ER6 (K values) of Compound IV were 21.7 + 1.7 nM (n = 3) and 15.2 ± 4.1 nM (n = 3), respectively. After binding to ER, Compound IV initiates a complex series of molecular events leading to the expression or repression of target genes involved with the drug response in a tissue-selective form. In transient transfection assays, Compound IV is an ERa and ERp agonist, with a demonstrated increased potency to stimulate ERa-mediated transcription activation as compared to that of ERp. While estradlol activates ERa and ERp with a selectivity greater than 5.1 times for ERa, Compound IV shows a selectivity of 49.0 times for ERa. Thus, Compound IV has a 9.7-fold relative selectivity in relative transactivation potency (normalized to estradlol values) for ERa over ERp. Furthermore, no antagonistic effects were observed in activation of estradlol-stimulated transcription (1 nM) with other nuclear hormone receptors, the actions of Compound IV are specific for ERa and ERp. Compound IV was classified for cross-reactivity against rat glucocorticoid receptor (GR), corticosteroid mineral receptor (MR), progesterone receptor (PR), androgen receptor (AR), and human farnesolde X receptor (FXR) isoforms. ), liver X receptor (LXR), peroxisome proliferator activated receptors (PPAR-α and PPAR-γ), and retlnoid X receptor (RXR-a) in both agonist and antagonist modes in

transcription activation assays. C om DTrestoHAA · η η reported no agonist or antagonist activity in any of these trials, supporting the conclusion that Compound IV does not functionally cross-react with these members of the nuclear hormone super-family.

EXAMPLE 7

Transactivation of selected compounds

Transactivation assays in agonist and antagonist modes were performed to identify whether the compound is an agonist, antagonist, or a partial.

Method

Rat estrogen receptors (ER-α and ER-β) were cloned from rat ovarian cDNA into a pCR3.1 plasmid vector base structure. Sequencing was performed to determine the absence of any mutation. HEK292 cells at 100,000 cells per well of a 24 well plate were plated in Dulbecco's Minimum Essential Medium (DMEM) + 5% charcoal-labeled fetal bovine serum (csFBS). Cells were transfected using Lipofectamine (Invitrogen, Carlsbad, CA) with 0.25 pg ERE-LUC, 0.02 pg CMV-LUC (renilla luciferase) and 12.5 ng rat ER-α or 25 ng rat ER-β. Cells were treated 24 hours after transfection with various concentrations of compounds or a combination of compounds and estradiol to

ΙΜ

101 determine antagonistic activity. Luciferase assays were performed 48 hours after transfection.

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Results

Classification of the compounds of this invention in the transactional system revealed that the compounds belonged to all three classes, ie agonists, antagonists and partial agonist. An example of an agonist and an antagonist is provided in Table 3. The results of transactivation coincided extremely well with the binding results for isoform selectivity.

Table 3 provides the ED values50 and IC50 of transactivation for some selected compounds of this invention.

TABLE 3. Transactivation (both agonist and antagonist) of selective compounds of this invention

<td></td><td colspan="2">COMPOUND</td><td></td><td></td><td rowspan="2">ec50</td><td rowspan="2">ic50</td><td rowspan="2">IC50</td>

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EXAMPLE 8

Testosterone Suppression in Macaque Monkeys

Two-year-old gonad macaque monkeys (n = 2) were housed during the study according to the USDA Guide Lines with free access to primate diet and water (except that there was fasting prior to oral dose administration) . Animals were given a once daily oral force feed dose of 30 mg / kg of the compound of formula IV in a Tween 80 / deionized water microemulsion vehicle for 7 consecutive days. Serum samples were withdrawn through a venipuncture prior to oral dose administration on days 1 (baseline), 3, 4, 5, 6, and 7. Total testosterone and androgens were quantified using a enzyme immunoassay method (EIA) combined with or without an HPLC method, respectively. After 6 days of treatment. with the compound of formula IV, the time-dependent reductions were apparent for testosterone and total androgens (testosterone / dihydrotestosterone). The compound of formula IV reduced testosterone levels by 58% and 64% in animal # 1 and animal # 3, respectively, with

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relation to baseline values (see solid lines tí'ir I a'-F'ngw »· 1; Table 4). Similarly, total androgen levels decreased to 56% in both # 1 and 3 # animals (see dotted lines in Figure 1; Table 4) compared to baseline values.

Consistent with estrogen re-augmentation of the ρ 11 axis uitari a-te st i cu I ar in males, these results demonstrated a robust pharmacological response to hormone suppression in serum- (testosterone and total androgens) in non-human primates. intact (macaque monkeys) after repeated oral doses (30 mg / kg) of the compound of formula IV.

TABLE 4. Testosterone and total androgen levels in the serum of intact male monkeys with daily oral administration of 30 mg / kg of the compound of formula IV (first dose on Day 0)

<td colspan="3">Testosterone</td><td colspan="2">Total Androgens</td>

<td colspan="3">Serum level (pg / ml)</td><td colspan="2">Serum level (pg / ml)</td>

<td>Day Π</td><td>Animal 1</td><td>Animal 3</td><td>Animal 1</td><td>Animal 3</td>

<td>or (baseline)</td><td> 1120</td><td> 617</td><td> 1868</td><td> 1643</td>

<td colspan="3">Reduction percentage from</td><td>Rec Percentage</td><td>uction from</td>

<td></td><td colspan="2">baseline</td><td>line d</td><td>e base</td>

<td>Day</td><td>Animal 1</td><td>Animal 3</td><td>Animal 1</td><td>Animal 3</td>

<td>(baseline)</td><td> 100</td><td></td><td></td><td></td>

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EXAMPLE 9 -

Suppression of LH and Testosterone Hormone Levels in Rats

An in vivo dose-response study was conducted to assess the effect of Compound IV on LH suppression in intact and orchiectomized (ORX) male rats. In intact and ORX animals, Compound IV at doses of> 10 mg / kg per day significantly suppressed LH levels when compared to respective controls. (The same pattern of suppression was observed at FSH levels). LH suppression resulted in robustly reduced testosterone levels to lower the limit of quantitation (BLOQ) which is 0.08 ng / ml and reduced prostate, seminal vesicles, and levator ani weight muscles and that these are organs highly dependent on androgen. In intact animals, dose dependent reductions in the weights of these target organs were observed with the weights of the seminal vesicles and the levator ani muscle at the level of castrated controls. Although the weights of the prostates were significantly reduced in intact animals, these values did not reach the level of castrated controls. The results are summarized in Table 6 below.

Materials and methods:

Male Sprague-Dawley rats, weighing approximately 200 g, were kept on a 12 hour light / dark cycle with food (rodent diet with 16% protein 2016 Teklad

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Global, Harían, Madison, Wl) and Agua HicponHi? ari lihitum. The animal protocol was reviewed and approved by the Institutionai Animal

Care and Use Committee of the University of Tennessee.

The test article for this study was weighed and dissolved in 10% DMSO (Fisher) diluted with PEG 300 (Acros Organics, NJ) to prepare dosage formulations. For this study, sixty (60) male Sprague-Dawley rats were randomly accommodated by body weight, and assigned to one of twelve treatment groups (n = 5 animals / group). Treatment groups are listed in Table 5. Animals were housed in groups of 2 to 3 animals per cage. Control groups (intact and orchiectomized (ORX)) were administered vehicle daily. Compound IV was administered through subcutaneous injection (200 pL) at doses of 0.3, 1, 3, 10, and 30 mg / kg / day to both the intact and ORX groups.

After a 14 day dosing regimen, animals were sacrificed under anesthesia (ketamine / xylazine, 87:13 mg / kg) and body weights were recorded. In addition, the ventral prostate, seminal vesicles, and levator ani muscle were removed, foreign tissue was cleaned, and individually weighed. Organ weights were normalized to body weights and expressed as a percentage of intact control. Blood was collected from the abdominal aorta under isoflurane anesthesia and allowed to clot. Serum was separated by centrifugation and stored at -80 ° C before determining the levels of

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serum hormone. Concentrations of serum luteinizing sister (LH) and serum follicle stimulating hormone (FSH) were determined through the Rat Pituitary Luminex Assay (Μ 11 ipo re, Blllerica, MA) according to the manufacturer's instructions. . The lower limit of quantification for this assay was 3.2 pg / ml for LH and 32 pg / ml for FSH. Testosterone was measured through a Testosterone EIA (Alpco Diagnostics, Salem, NH) with a BLOQ of 0.08 ng / ml. Serum hormone values below the lower limit of quantification (BLOQ) were omitted from the group media analysis. Therefore, the reported value for LH and T in the groups with samples, the BLOQ is greater than the actual value. This method of analysis provided the most conservative estimate of LH and T suppression. The Fisher Significant Minimum Difference test was used to compare individual dose groups with the intact vehicle and ORX control groups. Importance was defined a priori as a value P <0.05.

TABLE 5. Treatment Groups

<td>Group</td><td>Gonad State</td><td>Dose (mg / kg / day)</td><td>Test Item</td>

<td> 1,</td><td>Intact</td><td> -</td><td>Vehicle</td>

<td> 2</td><td>ORX</td><td> -</td><td>Vehicle</td>

<td> 3</td><td>Intact</td><td> 0.3</td><td>Compound IV</td>

<td> 4</td><td>Intact</td><td> 1</td><td>Compound IV</td>

<td> 5</td><td>Intact</td><td> 3</td><td>Compound IV</td>

<td> 6</td><td>Intact</td><td> 10</td><td>Compound IV</td>

<td> 7</td><td>Intact</td><td> - 30</td><td>Compound IV</td>

<td> 8</td><td>ORX</td><td> 0.3</td><td>Compound IV</td>

<td> 9</td><td>ORX</td><td> 1</td><td>Compound IV</td>

<td> 10</td><td>ORX</td><td> 3</td><td>Compound IV</td>

<td> 11</td><td>ORX</td><td> 10</td><td>Compound IV</td>

<td> 12</td><td>ORX</td><td> 30</td><td>Compound IV</td>

107

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Luteinizing Hormone Levels in Intact Rats and ORX (Table 6)

LH levels (± SD mean) in control groups of intact vehicle and ORX were 1.46 + 0.64 and 11.1 ± 3.9 ng / ml, respectively. It reduced the dose of Compound IV in intact animals, reaching statistically significant reductions with daily doses of> 3 mg / kg. LH levels in intact animals treated with Compound IV were 0.863 + 0.384, 0.704 + 0.530, 0.395 + 0.302, 0.226 + 0.165, and 0.236 ± 0.176 ng / ml, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively. LH levels in ORX males were also significantly reduced by treatment with Compound IV. In ORX animals, LH levels were 15.4 +.2.9, 13.5 +.2.2, 6.5 +.5.6, 0.425 and 0.135, and 0.368 + 0.119 ng / ml, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively. The results are presented graphically in Figure 10A.

Follicle Stimulating Hormone Levels in Intact Rats and ORX (Table 6)

Serum FSH levels in intact vehicle control groups and ORX were 20.9 ± 8.5 and 93.5 ± 13.8 ng / ml, respectively. In intact animals, the dose of Compound IV dependently reduced FSH levels with significant reductions observed at doses of> 10 mg / kg / day. The levels of

FSH in intact animals treated with Compound IV were

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17.3 ± 6.4, 15.7 + 7.3, 18.4 + 7.7, 9.2 + 4.0, and 6.3 ± 1 .K ...... h'Tí7hTt "'after doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively. 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, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively. The results are presented graphically in Figure 10B.

Testosterone Levels in Intact Rats and ORX

Testosterone levels in intact vehicle control groups were 2.4 ± 1.1 ng / ml. The lower limit of quantification for T was 0.08 ng / ml. Values less than 0.08 ng / ml were designated as Below the Limit of Quantitation (BLOQ). In Intact animals, the dose of the compound of formula IV reduced the levels of T with significant reductions observed at doses of> 3 mg / kg per day. Testosterone levels in intact animals treated with the compound of formula IV were 2.6 + 1.7, 1.6 ± 1.0, 0.7 ± 0.4, BLOQ, and BLOQ ng / ml, after doses of 0.3, 1, 3, 10, and 30 mg / kg per day, respectively. In ORX animals, T levels were LOCK for all groups treated with compound IV and the group treated with vehicle. Results for intact animals are graphically presented in Figure 10C (and Figure 2) (BLOQ values are plotted at the limit of quantification for graphic purposes).

The rapid and powerful suppression of serum testosterone in

109 Intact male rats were measured by administering Compound IV at doses of 3 mg / kg, 10 mg / kg and 300 mg / kg after 24 hours, 72 hours and 168 hours as presented in Figure 9.

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Organ Weights (Table 6)

The weights of the prostate, seminal vesicles, and the levator ani muscle were measured to confirm T. suppression. Organ weights (± SD mean) are presented in Figures 10D, 10E, and 10F, respectively. Dose dependent reductions in weight of prostate, seminal vesicles, or levator ani muscle were observed in intact animals treated with the

Compound IV. Prostate weights in intact animals were 84.0 + 19.2, 75.2 + 20.7, 68.2 + 8.1, 45.1 ± 20.0, and 43.6 + 8.8, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day . 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 doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively. The weights of the seminal vesicles in intact animals were 76.2 ± 7.8, 66.3 ± 27.2, 51.8 ± 28.5, 19.1 ± 7.0, t 17.9 ± 3.3, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively. Seminal vesicle weights in ORX animals were 12.2 + 1.3, 16.6 + 5.4, 16.5 + 4.8, 13.3 + 1.9, and 12.9 + 2.1, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day , respectively. Levator ani weights in intact animals were 86.9 ± 10.0, 82.1 ± 12.1, 65.2 + 4.4, 57.8 + 11.2, and 58.1 + 4.7, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively. The pesos

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of levator ani in ORX animals were 54.5 ± 6.6, 49.6 + 7.0, 53.6 + 10.0, 51.1 ± 4.9, and 49.2 + 4.2, after doses of 0.3, 1, 3, 10, and 30 mg / kg / day, respectively.

LH suppression data and organ weights are summarized in Table 6.

TABLE 6.

In vivo effects of the compound of formula IV on serum hormones and organ weight

<td>State of Gonads</td><td>Compound</td><td>Dose (mg / kg per day)</td><td></td><td>LH (ng / ml)</td><td>FSH (ng / ml)</td><td>Prostate (% of Intact)</td><td>Seminal Vesicles (% of Intact)</td><td>Ani's muscle of levator (% of Intact)</td>

<td>Intact</td><td>Vehicle</td><td> --</td><td>SD Means, medium</td><td>Ϊ465 0.642</td><td> 20.9b 8.49</td><td> 100.0b 28.6</td><td> 100.0b 13.4</td><td> 100.0b 4.97</td>

<td>ORX</td><td>Vehicle</td><td> -</td><td>SD Means, medium</td><td> 11.1to 3.87</td><td> 93.5to 13.8</td><td> 13.7to 2.56</td><td> 14.0to 2.93</td><td>58.8a 6.62</td>

<td>Intact</td><td>Compound IV</td><td> 0.3</td><td>SD Means, medium</td><td> 0.863b 0.384</td><td> 17.3b 6.44</td><td> 84.0b 19.2</td><td> 76.2ab 7.83</td><td> 86.9to? 10</td>

<td>Intact</td><td>Compound IV</td><td> 1</td><td>SD Means, medium</td><td> 0.704b 0.53</td><td> 15.7b 7.26</td><td> 75.2b 20.7</td><td> 66.3to? 27.2</td><td> 82.1to? 12.1</td>

<td>Intact</td><td>Compound IV</td><td> 3</td><td>SD Means, medium</td><td> 0.395to? 0.302</td><td> 18.4b 7.72</td><td> 68.2a, b 8.12</td><td> 518ab 28.5</td><td>65i2to 4.35</td>

<td>Intact</td><td>Compound IV</td><td> 10</td><td>SD Means, medium</td><td> 0.226to? 0.165</td><td> 9.25ab 3.97</td><td> 45.1to, b twenty</td><td> 19.1to 6.98</td><td> 57.8to 11.2</td>

<td>Intact</td><td>Compound IV</td><td> 30</td><td>SD Means, medium</td><td> 0.236to? 0.176</td><td> 6.25to? 1.82</td><td> 43.6to? 8.75</td><td> 17.9to 3.33</td><td> 58.1to 4.71</td>

<td>ORX</td><td>Compound IV</td><td> 0.3</td><td>SD Means, medium</td><td> 15.4to 2.94</td><td> 116to 17.2</td><td> 19.0to? 4.19</td><td> 12.2to' 1.31</td><td> 54.5to 6.56</td>

<td>ORX</td><td>Compound IV</td><td> 1</td><td>SD, Means, medium</td><td> 13.5to 2.18</td><td> 114to 22.3</td><td> 17.4to 3.4</td><td> 16.6to 5.36</td><td> 49.6to 7.04</td>

<td>ORX</td><td>Compound IV</td><td> 3</td><td>SD Means, medium</td><td> 6.5 5.63</td><td> 65.2to 31.9</td><td> 19.6to 6.67</td><td> 16.5to 4.82</td><td> 53.6to 10</td>

<td>ORX</td><td>Compound iv</td><td> 10</td><td>SD Means, medium</td><td> 0.425to? 0.135</td><td> 27.6b 8.16</td><td> 22.9to? 5.44</td><td> 13.3to 1.91</td><td> 51.1to 4.88</td>

<td>ORX</td><td>Compound IV</td><td> 30</td><td>SD Means, medium</td><td> 0.368ab 0119</td><td> 15.1b 4.11</td><td> 20.6to? 2.08</td><td> 12.9to 2.14</td><td> 49.2to? 4.21</td>

to P <0.05 against Intact Vehicle, b P <0.05 vs. ORX Vehicle

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EXAMPLE 10

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Recovery of Testosterone Levels After Suppression through Compound IV in Rats and Monkeys

The reversibility of chemical castration with Compound IV was studied.

Materials and methods:

Thirty-five (35) male Sprague-Dawley rats, weighing approximately 200 g, were kept on a 12-hour light / dark cycle with feeding (rodent diet with 16% protein 2016 Teklad Global, Harian, Madison , Wl) and water ad libitum. The animal protocol was reviewed and approved by the Institutionai Animal Care and Use Committee of the University of Tennessee.

The test article for this study was weighed and dissolved in PEG 300 (100%) (Acros Organize, NJ) to prepare the appropriate dosage formulations. Animals were randomly assigned to one of the ten treatment groups (n = 5 animals / group). Treatment groups are listed in Table 7. Animals were housed in groups of 2 to 3 animals per cage. Group 1 was sacrificed at the start of the study (Day 1) for the determination of baseline testosterone levels in intact animals. Groups 2-7 received daily doses of 1, 3, or mg / kg through oral forced feeding (~ 200 uL) for three days. Groups 2, 3, and 4 were euthanized on Day 4 to measure maximum testosterone suppression. Groups 5, 6, and 7 were left ft · ^

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112

He will recover for 14 days with a drug-free washout period.

TABLE 7. Treatment groups

<td>Group</td><td>Compound IV PO Dose</td><td>Treatment</td>

<td>Group 1</td><td> —</td><td>Baseline</td>

<td>Group 2</td><td>1 mg / kg for 3 days</td><td>No Recovery</td>

<td>Group number 3</td><td>3 mg / kg for 3 days</td><td>No Recovery</td>

<td>Group 4</td><td>30 mg / kg for 3 days</td><td>No Recovery</td>

<td>Team 5</td><td>1 mg / kg for 3 days</td><td>14 days recovery</td>

<td>Group 6</td><td>3 mg / kg for 3 days</td><td>14 days recovery</td>

<td>Group 7</td><td>30 mg / kg for 3 days</td><td>14 days recovery</td>

Results:

Serum testosterone levels in Intact rats were 6.4 ± 3.1 ng / ml (± SD mean) at baseline. Compound IV administered at doses of 3 and 30 mg / kg for three days significantly suppressed serum testosterone levels at 1.47 ± 0.26 and 1.62 ± 0.49 ng / ml, respectively. We did not observe any significant deletion in animals that received 1 mg / kg of Compound IV for 3 days. More importantly, serum testosterone levels were 3.3 ± 1.92, 3.00 ± 1.06, and 3.8 ±

1.72 in animals receiving 1, 3, or 30 mg / kg, respectively, of Compound IV for three days when

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INDUSTRIAL measured after a 14-day recovery period, and there were no statistically significant differences from baseline serum testosterone concentrations in intact rats as shown in Figure 23.

This study confirms previous results showing that the

Compound IV rapidly suppresses serum testosterone levels in intact male rats. Suppression of serum testosterone levels was observed in dose groups receiving> 3 mg / kg / day for 3 days. No significant reduction in serum testosterone was observed with the 1 mg / kg dose group. However, during the 14 days of recovery, testosterone levels returned to the level of intact controls. This study shows that pharmacological castration by Compound IV is reversible in rats.

The effect of Compound IV on the suppression and recovery of testosterone levels in Intact male monkeys was evaluated in conjunction with an oral pharmacokinetic study. Three treatment natural male macaque monkeys (age 2 to 3 years) are treated. Compound IV was administered at 30 mg / kg daily through oral force feeding for 7 consecutive days. Blood samples were taken and divided into serum and plasma for quantitative measurements of testosterone and Compound IV, respectively. The results show that daily oral doses of Compound IV significantly reduced circulating androgen levels (primarily testosterone and dlhydrotestosterone) in all three monkeys.

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male by up to 47% compared to baseline levels (1591 ± 72.5, 997 ± 104, and 852 ± 136 ng / ml, respectively for baseline, Day 2 and Day 6 of treatment [± SEM mean]). After an 18-day drug-free recovery period, androgen levels returned to normal, and were not significantly different from pre-treatment baseline levels (1757.7 ± 369.5 ng / ml after recovery) .

EXAMPLE 11

Bone Conservation Despite LH and Testosterone Reduction in Rats (Table 8)

The effect of the compound of formula IV was studied in. bone treatment. The fully orally administered compound of formula IV prevented bone loss associated with LH suppression in intact male rats. S induced a significant reduction in LH through the compound of formula IV in intact animals at dose levels of> 10 mg / kg per day. Although at 1 mg / kg per day, the compound of formula IV did not significantly reduce LH, significant reductions in prostate, seminal vesicles, and levator ani muscle were evident at this dose, indicating that the reduction in circulating testosterone was physiologically relevant in these androgen sensitive organs. However, 1 mg / kg per day of the compound of formula IV kept the trabecular bone volume (measured in the distal femur) at the level of intact controls. When administered at doses of 10 and 30

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mg / kg per day, the compound of formula IV increased bone volume in the distal femur significantly above that of intact controls. These data show that compound IV increased trabecular bone mineral density (BND) and percent bone volume at a dose level that reduces LH levels in Intact rats. The data from this study are presented in the

Table 8.

TABLE 8. In Vivo Effects of Compound IV on Rat Bone, Organ, and Hormone Parameters 10

<td>State of Gonads</td><td>Compound</td><td>Dose (mg / kg by day)</td><td></td><td>Density mining! that is (g / cm3)</td><td>Percentage of bone volume (BWTV) (%)</td><td>Prostate (%of Intact)</td><td>Vesicles seminal (% of Intact)</td><td>An¡ muscle of levator (% of Intact)</td><td>FSH (ng / ml)</td><td>LH (ng / ml)</td>

<td>Intact</td><td>Vehicle</td><td> -</td><td>SD Means, medium</td><td> 0.274 0.033</td><td> 20.2 3.57</td><td> 100.0 11.1</td><td> 100.0 15.9</td><td> 100.0 11.6</td><td> 993 2.94</td><td> 0.781 0.263</td>

<td>ORX</td><td>Vehicle</td><td> -</td><td>SD. Means, medium</td><td> 0.224to 0.025</td><td> 15.4to 2.6</td><td> 14.8to 4.08</td><td> 10.3to 0.767</td><td> 59.4to 7.26</td><td> 117to 40.2</td><td> 22.0to 5.81</td>

<td rowspan="2">Intact</td><td>Compound</td><td rowspan="2"> 1</td><td>SD</td><td> 0.273b</td><td> 20.0b</td><td> 69.2to</td><td> 44.6a, b</td><td> 80.0ab</td><td> 14.1ab</td><td> 0820”</td>

<td>IV</td><td>Means, medium</td><td> 0.04</td><td> 4.08</td><td> 13.5</td><td> 15.7</td><td> 6.69</td><td> 4.07</td><td> 0.392</td>

<td>Intact</td><td>Compound IV</td><td> 10</td><td>SD Means, medium</td><td> 0.326to'6 0.048</td><td> 25.9to” 4.76</td><td> 30.7to” 12.4</td><td> 12.8ab 0.88S</td><td> 58.1to 9.68</td><td> 5.48’” 1.97</td><td> 0.060to” 0.092</td>

<td rowspan="2">Intact</td><td>Compound</td><td rowspan="2"> 30</td><td>SD</td><td> 0.326to'”</td><td> 25.5ab</td><td> 30.1to'6</td><td> 14.4to”</td><td> 56.1to</td><td> 6.32to”</td><td> 0.078to”</td>

<td>IV</td><td>Means, medium</td><td> 0.046</td><td> 4.49</td><td> 17.4</td><td> 1.45</td><td> 4.67</td><td> 3.4</td><td> 0.114</td>

toP <0.05 against Intact Vehicle. b P <0.05 vs. ORX Vehicle

EXAMPLE 12 /

Effects on the enzymatic activity of 1 7p-hydroxysteroid dehydrogenase 5 (1 7p-HSD5)

Members of the HSD family were involved in the conversion of circulating spheroids. 17P-HSD5 converts androstenedione to

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INDUSTRIAL 116 'testosterone and estrone to estradiol. In addition, he was also involved in prostaglandin synthesis. Here the ability of some selected compounds of this invention to inhibit the activity of 17p-HSD5 was demonstrated.

Method

Human 17p-HSD5 was cloned into the pGEX 4t1 vector and a purified protein was prepared. The purified protein was incubated with the representative compound of this invention,14C androstenedione and NADPH in an appropriate pH regulator. Synthesized testosterone was extracted using ethyl acetate, air dried, splashed, and run on a thin layer chromatography (TLC) plate. TLC was exposed to phospho-imaging and the intensity of the testosterone band was quantified. Indomethacin was used as a positive control (LHRH agonist).

Results

Compound IV was tested and had a partial inhibitory effect on the activity of the 173-HSD5 enzyme. The positive control (LHRH agonist), indomethacin, was expected to exhibit strong inhibition of this enzyme, as presented in Figure 3.

EXAMPLE 13

Toxicity studies

A study was conducted to compare the thrombotic potential

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MEXICAN INSTITUTE of Compound IV and diethylstylbestro! (D ES, positive control) using the human platelet aggregation assay. Blood from healthy male donors was used in the study although male donors are intended for the treatment population for Compound IV (LH suppression). Platelet rich plasma was pre-incubated with estradiol (E2), compound IV, or vehicle for 30 seconds, and then thrombin (0.3 units) was added to indicate platelet aggregation. The study results show that pre-incubation with DES increased thrombin-induced platelet aggregation by approximately 10-fold. However, Compound IV and estradiol reduced aggregation in platelet rich plasma. These data demonstrate that Compound IV reduced the reactivity of human platelets in vitro, compared to DES, and suggests that Compound IV may have a lower boembolic trom potential than DES (Figure 4).

EXAMPLE 14

Effect of Compound IV on Hot Flashes

A study was conducted to investigate the effect of Compound IV on hot flashes using the morphine dependent rat model (MD model), which was developed by Simpkins et al. (1 983) and was shown to have several similarities to the hot flash of menopause. In addition to the similarities to the human condition, this experimental animal model has a short spin around time which makes it a useful high ranking tool.

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Mexican werrnrro or the womeoap INDUSTRIAL production to identify compounds that can alleviate IW? Vaso-motor symptoms using the tail skin temperature (TST). TSTTA-40 probes (Data Sciences International, MN) were placed at the base of the tails and baseline temperatures were obtained for 15 minutes. After 15 minutes, the animals were treated with naloxone (1 mg / kg, SQ) to reverse the effects of morphine. Tail temperature (TST) was measured for one hour after naloxone treatment at a sampling rate of 5 seconds throughout the experiment. After data acquisition, the average temperature movement recorded every 60 seconds for each animal was calculated and further analyzed. The baseline temperature was calculated as the average temperature acquired during the 15 minutes preceding naloxone administration. Area under the curve (AUC) was calculated by subtracting all values after naloxone administration from baseline using a linear trapezoid method. .

Compound IV attenuated hot flashes in the morphine withdrawal model (see Figure 13) with the best results obtained at a concentration of 10 mg of Compound IV. 1 7β E2 at 5 mg / kg in 100% DMSO was used.

EXAMPLE 15

Compound IV vs. DES in Rats

Before the introduction of LHRH antagonists,

I

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY castration testosterone levels increasing the activity of estrogen in the pituitary through estrogens, mainly diethylstilbestrol (DES). DES was equally effective as LHRH agonists in suppressing testosterone at castration levels. Patients treated with DES did not have hot flashes or bone loss, but they did have gynecomastia at higher rates than ADT with LHRH agonists.

Unfortunately, pure, highly potent estrogens, such as DES and estradiol, are generally associated with a high risk of cardiovascular complications and severe bolus thromboem, which have limited their clinical use. The hypothesis has been made, but not proven, that the increased risk of venous thromboembolic complications with DES is due to its cross-reactivity with other hormone receptors. In vitro studies with human platelets showed that Compound IV had much lower procoagulant activity than DES. In this way, Compound IV, a selective ER-alpha agonist, can deliver the benefits of DES to prostate cancer and also deliver the benefits of an LHRH agonist without causing osteoporosis or adverse lipid profiles.

Compound IV is as effective as DES in reducing prostate size in rats (Figure 11 A) and exhibits a moderate increase in prostate size in ORX rats (Figure 11 B).

The differences between DES and Compound IV are presented in

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120 Figures 12A-12C, where DES cross-reacts with the glucocorticoid receptor (GR) (Figures 12A) and the androgen receptor (AR) (Figure 12B) while in Compound IV it does not. Furthermore, DES antagonized estrogen-related receptor transactivation (ERR), while Compound IV did not. Compound IV failed to cross-react with any of the three ERR isoforms (ERR-a, EER-β and ERRy) as shown in Figure 12C.

EXAMPLE 16

Monkey Toxicity Study - 90 Days

Macaque monkeys were obtained from the island's breeding colony

Mauricio. The prospective study was designed as a 36-week oral pharmacological and toxicological evaluation of Compound IV and positive control (LHRH agonist) in male macaque monkeys with a provisional period of 13 weeks. A total of 39 sexually mature male monkeys, 5 to 8 years of age, were randomly assigned to five groups before the start of treatment. Groups included: 1) vehicle control, 2) 1 mg / kg Compound IV, 3) 10 mg / kg Compound IV, 4) 100 mg / kg Compound IV, and 5) positive control (LHRH agonist). The drug was administered orally through lateral cage administration once daily for 39 weeks with the vehicle control article (Tween 80 / PRANG ™) for Groups 1 and 5, or Compound IV in the vehicle for Groups 2, 3, and 4. The dose levels of Compound IV were 1,

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10, and 100 mg / kg / day for Groups 2, 3, and 4, respectively. Oral doses were delivered in a dose volume of 10 ml / kg as calculated based on the most recent available body weight for each animal (Figure 14). Animals in Group 5 also received a once daily subcutaneous injection of positive control (LHRH agonist / 0.02 my constant volume) during the 39 week study period. General appearance and clinical signs were observed and recorded daily. Routine evaluations were performed and other study investigations were selected as indicated in the study protocol. Parameter selection includes, but is not limited to, testosterone, prostate specific antigen (PSA), and prostate volume and weight.

Total testosterone and PSA levels were quantified in serum samples (following the standard procedure) using an enzyme immunoassay (EIA) and chemiluminescence immunoassay (LIA, ALPCO Diagnostics, Salem NH) method, respectively. Blood samples for testosterone evaluations were taken from all animals (in the fasted state) at a baseline (i.e. before starting treatment) and on Days 1, 3, 7, 14, 28, 64, and 90. Blood samples for PSA determinations were taken from all animals (in a fasted state) at baseline and during Week 6. For discussion purposes, results were calculated for samples with concentrations below the limit of quantification ( BLQ) for

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Testosterone and PSA tests, like half of it ΙΙΓΤΙΙΤ? Iliftíi iui dela quantification (LLOQ) of the trial, and were considered as “Estimated final concentrations”. Data in Tables 9 through are presented as “Quantifiable Concentrations Only” (ie excludes BLQ values) in addition to “Estimated Final Concentrations” (ie displayed with BLQ result included as 1/2 LLOQ of assay) . Prostate volume was measured in live animals under anesthesia using a transrectal ultrasound (TRUS) procedure at baseline and Week 6. The width and height of the prostate were recorded. Prostate volumes were calculated as width x width xp / 6 and normalized to body weight. The wet weight of the prostate was recorded with an autopsy after cutting the fat-free and foreign tissue.

Results and Discussion:

Serum testosterone levels are presented in Figure 15 and Tables 9 through 12. At a baseline, testosterone levels for all monkeys in the study were on the normal scale for sexually mature adult macaque monkeys. However, testosterone levels were significantly reduced in monkeys receiving Compound IV at 100 mg / kg / day in monkeys treated with positive control (LHRH agonist). Testosterone levels in the positive control group (LHRH agonist) illustrated a biphasic change, with a

Say THE PROPERTY Λ, .. INDUSTRIAL * «,

123 significant initial increase (i.e. widening) of 47.4% and 547% (p <0.01) on Days 1 and 3, respectively, followed by reductions of 3.6%, 67%, 73%, 83%, and 85% in the Days 7, 14, 28, 64 and 90 (see Figure 15 and Tables 9 to 12). No similar widening was observed for any animal treated with Compound IV even at the highest dose level (ie 100 mg / kg / day). The dose and duration of treatment were important for the pharmacological action of Compound IV, where doses of 100 mg / kg / day suppressed serum testosterone by 60%, 51%, 42%, 79% and

92% on Days 3, 7, 14, 28 and 64, respectively, in relation to the baseline value (see Figure 15 and Tables 9 and 10). After 90 days of treatment with 100 mg / kg / day of Compound IV, the testosterone level in 6 of 10 monkeys in Group 4 was reduced to concentrations below the assay quantification limit (refer to Table 11). The mean serum testosterone level in Group 4 monkeys was reduced by 95% compared to respective baseline values ("Estimated Final Concentrations", 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 for Day 90, Compound IV at a concentration of 100 mg / kg / day reduced the serum testosterone level to levels significantly lower than the positive control (LHRH agonist) (p = 0.013).

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TABLE 9. Average testosterone levels (mg / ml) in intact male monkeys after daily oral administration of

Compound IV; @ Estimated final concentrations

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<td rowspan="2">Day</td><td colspan="3" rowspan="2">Vehicle Control</td><td colspan="3">Compound IV</td><td colspan="3" rowspan="2">Compound IV 10 mg / kg</td><td colspan="3" rowspan="2">Compound IV 100 mg / kg</td><td colspan="3" rowspan="2">Positive Control (LHRH agonist)</td>

<td></td><td>Means, medium</td><td>SEM</td><td>N</td><td>Means, medium</td><td>SEM</td><td>N</td><td>Means, medium</td><td>SEM</td><td>N</td><td>Means, medium</td><td>SEM</td><td>N</td><td>Means, medium</td><td>SEM</td><td>N</td>

Testosterone Test LLOQ = 0.246 ng / ml; @ BLOQ values were calculated as 0.123 ng / ml, half of LLOQ.

*: Statistically important (p <0.05) Compound IV 10 mg / kg against Vehicle Control #; Statistically important (p <0.05) Positive control (LHRH agonist) against Vehicle Control $: Statistically important (p <0.05) Positive control (LHRH agonist) against Compound IV 100 mg / kg &. • .fUW

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TABLE 10. Percentage (%) of change serum mean testosterone levels compared to baseline;

@ Estimated final concentrations

<td>Day</td><td>Control of Vehicle</td><td>Compound IV 1 mg / kg</td><td>Compound IV 10 mg / kg</td><td>Compound IV 100 mg / kg</td><td>Control Positive</td>

Testosterone test LLOQ = 0.246 ng / ml; @ BLQ values were calculated as 0.123 ng / ml, half of LLOQ.

TABLE 11. Serum mean testosterone levels (ng / ml) in intact male monkeys after daily oral administration of Compound IV; xQuantifiable concentrations only

<td>Day</td><td colspan="3">Control of Vehicle</td><td colspan="3">Compound IV 1 mgíkg</td><td colspan="3">Compound IV 10 mg / kg</td><td colspan="3">Compound IV 100 mg / kg</td><td colspan="3">Positive Control (LHRH agonist)</td>

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Testosterone Test LLOQ = 0.246 ng / ml; λ LOCK values were excluded.

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TABLE 12. Percentage (%) of change in mean testosterone levels compared to baseline; TOQuantifiable concentrations only

<td>Day</td><td>Control of Vehicle</td><td>Compound IV 1 mg / kg</td><td>Compound IV 10 mg / kg</td><td>Compound IV 100 mg / kg</td><td>Control Positive</td>

LLOQ Testosterone Test = 0.246 ng / ml; λ values are excluded

BLQ.

Serum PSA levels were also significantly suppressed by Compound IV within four weeks from the start of treatment. PSA reductions of 69% and 87% (on average) were observed for the monkeys receiving Compound IV at 10 mg / kg and 100 mg / kg for 4 weeks, while PSA levels were reduced by 60% in the positive control group (LHRH agonist) (Figure 16 and Tables 1 3-1 6).

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TABLE 13. Average serum PSA levels (ng / ml) in intact male monkeys after daily oral administration of the

Compound IV; ® Estimated final concentrations

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<td></td><td colspan="3">Control of Vehicle</td><td colspan="3">Compound IV 1 mg / kg</td><td colspan="3">Compound IV 10 mg / kg</td><td colspan="3">Compound IV 100 mg / kg</td><td colspan="3">Positive Control (agonist of LHRH)</td>

LLOQ PSA assay = 0.0575 ng / ml; @BLQ values were calculated as 0.02875 ng / ml, half of LLOQ.

*: Statistically important (p <0.05) Compound IV 10 mg / kg against Vehicle Control &: Statistically important (p <0.05) Compound IV 100 mg / kg against Vehicle Control #: Statistically important (p <0.05) Positive control ( LHRH agonist) vs. Vehicle Control $; Statistically Important (p <0.05) Positive control (LHRH agonist) against Compound IV 100 mg / kg

<td></td><td>Vehicle Control</td><td>Compound IV 1 mg / kg</td><td>Compound IV 10 mg / kg</td><td>Compound IV 100 mg / kg</td><td>Positive Control (LHRH agonist)</td>

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PSA test LLOQ = 0.0575 ng / ml; @Values of

BLQ as 0.2875 ng / ml, half the LLOQ.

TABLE 15. Average serum PSA levels (ng / ml) in intact male monkeys after oral administration of Compound IC; xQuantifiable concentrations only

PSA Assay LLOQ = 0.0575 ng / ml; TOBLQ values are excluded in this table.

TABLE 16. Percentage (%) of change in mean PSA levels compared to baseline; xQuantifiable concentrations only

<td></td><td>Control of Vehicle</td><td>Compound IV 1 mg / kg</td><td>Compound IV 10 mg / kg</td><td>Compound IV 100 mg / kg</td><td>Positive Control (LHRH agonist)</td>

PSA test LLOQ = 0.0575 ng / ml; zValues of

BLQ in this box.

Prostate volumes were measured through TRUS periodically throughout the study. The results obtained

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MKXICANO INDUSTRIAL PROPERTY INSTITUTE> j jfjj after six weeks of treatment demonstrated a potent effect of Compound IV and positive control (LHRH agonist) in monkey prostate. Compound IV significantly suppressed prostate volumes by 25% and 45% at dose levels of 10 mg / kg and 100 mg / kg, respectively, while prostate volumes were reduced by 28% in the control group. positive (LHRH agonist) (Figure 17 and Tables 17 and 18).

TABLE 17. Average Prostate Volumes (Ratio) in Male Monkeys After Daily Oral Administration of Compound IV

<td></td><td colspan="3">Vehicle Control</td><td colspan="3">Compound IV 1 mg / kg</td><td colspan="3">Compound IV 10 mg / kg</td><td colspan="3">Compound IV 100 mg / kg</td><td colspan="3">Positive Control (LHRH agonist)</td>

TABLE 18. Percentage (%) of change in average prostate volumes compared to baseline

<td></td><td>Control of Vehicle</td><td>Compound IV 1 mg / kg</td><td>Compound IV 10 mg / kg</td><td>Compound IV 100 mg / kg</td><td>Positive Control (LHRH agonist)</td>

Compound IV related reductions in prostate volume were confirmed through evaluation of prostate weight at necropsy. After thirteen weeks of

AND

130 treatment, Compound IV significantly reduced mean prostate weights by 24% and 21% in animals receiving 10 and

100 mg / kg / day, respectively (Figure 18B and Tables 19 and 20).

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TABLE 19. Average Prostate Weights (grams) at Necropsy of Monkeys with Daily Oral Administration of Compound IV

<td rowspan="2"></td><td colspan="3">Control of Vehicle</td><td colspan="3">Compound IV 1 mg / kg</td><td colspan="3">Compound IV 10 mg / kg</td><td colspan="3">Compound IV 100 mg / kg</td>

<td>Means, medium</td><td>SEM</td><td>N</td><td>Means, medium</td><td>SEM</td><td>N</td><td>Means, medium</td><td>SEM</td><td>N</td><td>Means, medium</td><td>SEM</td><td>N</td>

TABLE 20. Percentage (%) of change of average prostate weights compared to baseline

<td></td><td>Control</td><td>Compound IV 1 mg / kg</td><td>Compound IV 10 mg / kg</td><td>Compound IV 100 mg / kg</td>

No apparent effects on platelet aggregation, prothrombin time (PT), or activated partial thromboplastin time (APTT) were observed.

EXAMPLE 17

Compound IV Studies in Humans

A study was conducted to determine the effect of Compound IV in male humans. Twelve subjects per group were examined a

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doses of 1, 300, 600 and 1,000 mg of Compound IV. Table 21 presents the mean change in levels of LH, serum PSA, free testosterone, and total testosterone in men by administering Compound IV at doses of 1, 300, 600, and 1,000 mg. Dose-dependent mean total testosterone levels (nmoles / l) were measured in humans over a period between days 111 (Figure 19). Total testosterone level decreased by 51.9% and 47.9% at doses of 600 mg and 1000 mg, respectively.

Average dose-dependent LH levels (IUZI) in humans were measured over a period of days 1-10 (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 1,000 mg, respectively.

Dose-dependent free testosterone levels (pg / ml) were measured in humans over a period of days 1-10 (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.

Dose-dependent mean PSA levels (pg / L) in humans were measured over a period of days 1-10 (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 1,000 mg, respectively. No changes were observed for 10 and 30 mg doses.

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TABLE 21. Average change from baseline

<td>PSA serum</td><td> -9.2%</td><td> -24.4%</td><td> -27.5%</td><td> -29.9%</td>

<td>Free Testosterone</td><td> ' -17.0%</td><td> -18.5%</td><td> -72.7%</td><td> -53.2%</td>

<td>Total Testosterone</td><td> 3.9%</td><td> 7.3%</td><td> -51.9%</td><td> -47.9%</td>

EXAMPLE 18

Bioavailability of Compound IV

Compound IV was rapidly absorbed after oral dosing in rats, dogs and monkeys. The oral bioavailability of Compound IV in rats ranged from 6% to 25% depending on the formulation where the dose was administered. Formulations using polyethylene glycol 300 (PEG 300) generally produced higher exposures than the ml microemulsions prepared in Tween 80 diluted in de-flaked water. In dogs, visual inspection of the plasma-time concentration profiles suggested that Compound IV underwent enterohepatic recirculation as evidenced by a second peak in the terminal phase. Importantly, in dogs, exposure in the oral PEG 300 30 mg / kg male dose group exceeded the exposure necessary to produce the maximum effect on prostate reduction in the rat LH suppression model . In monkeys, preliminary pharmacokinetic studies suggested that oral bioavailability in this species approximates or exceeds that in dogs, as evidenced by concentrations of Compound IV in plasma and suppression of

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serum testosterone over a period of seven days. As a whole, these data suggest that sufficient oral exposure can be obtained in two non-rodent animal species to produce the desired pharmacological effect (based on AUC data). In addition, endocrine data in rats and monkeys suggest that the pharmacokinetic effects of Compound IV are reversible (i.e., that serum testosterone concentrations return to baseline or normal levels when Compound IV treatment is stopped).

EXAMPLE 19

Pharmacokinetics of Compound IV

Preliminary data from metabolism studies, in vitro (mouse, rat, dog, monkey and humans) and in vivo (rat) suggest that the conjugation of Compound IV, its hydroxylated metabolite (s) and its N-dealkylated metabolite they contribute to the total disposal of Compound IV in animals and humans. The results of the comparison between species, although only qualitative, show that the total metabolite profiles of the non-clinical species adequately reflect the profile generated in human liver mlrosomes. Based on these studies, rat and dog are appropriate rodent and non-rodent species, respectively, for pharmacological and toxicological evaluations. In vitro studies show that Compound IV does not induce relevant CYP450 isoforms (CYP1A2, CYP2B6, or CYP3A4), and does not inhibit CYP1A2,

134

CYP2C19, CYP2D6, or CYP3A4 / 5 at concentrations of <30 μΜ.

CYP2C9 is inhibited by Compound IV but only at high concentrations (K¡ = 8 µΜ), and potential pharmacokinetic drug-drug interactions are considered remote.

EXAMPLE 20

Biological Activity of Compound IV

Compound IV exerts little or no inhibitory effects in vitro (IC50> 300 μΜ) in the hERG channel. The dose-dependent compound reduced APD50 and APD90 at concentrations of 10 and 100 µΜ in isolated canine Purkinje fibers, in vitro. However, Compound IV did not affect hemodynamic or cardiac function (blood pressure), heart rate, EKG morphology, or QT intervals) in telemetered dogs at any dose (up to 300 mg / kg). Neuropharmacological or pulmonary effects were not observed. No significant effects on renal function were noted with a single oral dose of up to 30 mg / kg of Compound IV. Increased output of urine volume and urinary excretion of potassium and chlorine were only observed at the highest dose tested (100 mg / kg). Administration of Compound IV at doses of 30 to 300 mg / kg in rats produced a significant increase in peristalsis, and oral administration of Compound IV at 30 mg / kg in rats produced a significant increase in gastrointestinal motility and gastric acidity (probably not due to the effects on smooth muscle).

MEXICAN INSTITUTE

OF THE PROPERTY

INDUSTRIAL

135

Compound IV was not mutagenic and did not induce numerical or structural chromosomal aberrations at concentrations up to

200 μΜ in human peripheral blood lymphocytes, in vitro. Compound IV was well tolerated by rats and dogs after individual and repeated oral administration (up to 28 days). No pathological changes were observed in the kidney, liver, heart, and other related non-target organs. There were no serious physical signs, effects on body weight, clinical pathology changes, ophthalmological, electrocardiographic, or histopathological changes associated with oral administration of Compound IV to female or male dogs for up to 28 days.

Although certain aspects of the invention have been illustrated and described herein, many modifications, substitutions, changes, or equivalents will now occur to those skilled in the art. Therefore, it should be understood that the appended claims are intended to cover such modifications and changes to fall within the true spirit of the Invention.

136

IMPI Mexican institute OF INDUSTRIAL PROPERTY

Contents161

97 members in 14 offices

This recordMember, by publication datePriority claim4 more offices are in the list below

Priority claims19

Priority claims
Document	Office	Kind	Date
15470709	United States of America	P	2009-02-23
15470709	United States of America	P	2009-02-23
61154707	United States of America	–	2009-02-23
16898309	United States of America	P	2009-04-14
16898309	United States of America	P	2009-04-14
61168983	United States of America	–	2009-04-14
26166909	United States of America	P	2009-11-16
26166909	United States of America	P	2009-11-16
61261669	United States of America	–	2009-11-16
2010025032	United States of America	W	2010-02-23
2010025032	United States of America	W	2010-02-23
61154707	–	–	–
61168983	–	–	–
61261669	–	–	–
PCTUS2010025032	–	–	–
US20090154707P	–	–	–
US20090168983P	–	–	–
US20090261669P	–	–	–
WO2010US25032	–	–	–

Members97

Family members, oldest first
Document	Office	Kind
AU2006318400A1	Australia	A1	2007-05-31
CA2631331A1	Canada	A1	2007-05-31
CA2824517A1	Canada	A1	2007-05-31
WO2007062230A2	World Intellectual Property Organization (WIPO)	A2	2007-05-31
US2007135407A1	United States of America	A1	2007-06-14
WO2007062230A3	World Intellectual Property Organization (WIPO)	A3	2007-09-20
US2007265296A1	United States of America	A1	2007-11-15
EP1954670A2	European Patent Office (EPO)	A2	2008-08-13
MX2008006885A	Mexico	A	2008-10-20
WO2008130571A1	World Intellectual Property Organization (WIPO)	A1	2008-10-30
IL191803A0	Israel	A0	2008-12-29
EA200801461A1	Eurasian Patent Organization (EAPO)	A1	2008-12-30
CN101336227A	China	A	2008-12-31
US2009062341A1	United States of America	A1	2009-03-05
KR20100014228A	Republic of Korea	A	2010-02-10
JP2010524939A	Japan	A	2010-07-22
CA2753436A1	Canada	A1	2010-08-26
WO2010096801A1	World Intellectual Property Organization (WIPO)	A1	2010-08-26
US2010267773A1	United States of America	A1	2010-10-21
EP1954670A4	European Patent Office (EPO)	A4	2011-04-27
CA2790703A1	Canada	A1	2011-09-01
WO2011106317A1	World Intellectual Property Organization (WIPO)	A1	2011-09-01
AU2010215809A1	Australia	A1	2011-09-22
BRPI0620520A2	Brazil	A2	2011-11-16
IL214805A0	Israel	A0	2011-11-30
KR20110131227A	Republic of Korea	A	2011-12-06
EP2398322A1	European Patent Office (EPO)	A1	2011-12-28
CN102351732A	China	A	2012-02-15
US2012077845A1	United States of America	A1	2012-03-29
AU2006318400B2	Australia	B2	2012-04-05
CN102413692A	China	A	2012-04-11
US8158828B2	United States of America	B2	2012-04-17
MX2011008879A	Mexico	A	2012-04-30
EP2455362A1	European Patent Office (EPO)	A1	2012-05-23
US2012157539A1	United States of America	A1	2012-06-21
IL219590A0	Israel	A0	2012-06-28
JP2012518654A	Japan	A	2012-08-16
AU2011221246A1	Australia	A1	2012-09-13
CN102702013A	China	A	2012-10-03
EP2398322A4	European Patent Office (EPO)	A4	2012-12-05
CN102858154A	China	A	2013-01-02
EP2538780A1	European Patent Office (EPO)	A1	2013-01-02
KR20130000398A	Republic of Korea	A	2013-01-02
AU2006318400C1	Australia	C1	2013-01-17
EA201201021A1	Eurasian Patent Organization (EAPO)	A1	2013-01-30
RU2011137324A	Russian Federation	A	2013-03-27
CA2845890A1	Canada	A1	2013-03-28
WO2013043304A1	World Intellectual Property Organization (WIPO)	A1	2013-03-28
AU2012312902A1	Australia	A1	2013-05-23
EA018066B1	Eurasian Patent Organization (EAPO)	B1	2013-05-30
JP2013520445A	Japan	A	2013-06-06
KR20130101146A	Republic of Korea	A	2013-09-12
US8546451B2	United States of America	B2	2013-10-01
US8637706B2	United States of America	B2	2014-01-28
US2014057946A1	United States of America	A1	2014-02-27
US2014057985A1	United States of America	A1	2014-02-27
IL191803A	Israel	A	2014-03-31
IL231070A0	Israel	A0	2014-03-31
WO2013043304A9	World Intellectual Property Organization (WIPO)	A9	2014-04-24
EP2538780A4	European Patent Office (EPO)	A4	2014-04-30
KR20140064906A	Republic of Korea	A	2014-05-28
US2014171468A1	United States of America	A1	2014-06-19
EP2747562A1	European Patent Office (EPO)	A1	2014-07-02
US2014187641A1	United States of America	A1	2014-07-03
CN103957706A	China	A	2014-07-30
JP2014166996A	Japan	A	2014-09-11
JP2014524479A	Japan	A	2014-09-22
MX2014002105A	Mexico	A	2014-09-25
JP5611991B2	Japan	B2	2014-10-22
CN102351732B	China	B	2014-10-29
KR101458539B1	Republic of Korea	B1	2014-11-10
KR20150001847A	Republic of Korea	A	2015-01-06
CN102413692B	China	B	2015-01-07
RU2543339C2	Russian Federation	C2	2015-02-27
US2015087712A1	United States of America	A1	2015-03-26
IL219590A	Israel	A	2015-03-31
EP2747562A4	European Patent Office (EPO)	A4	2015-05-06
IN1959DEN2014A	India	A	2015-05-15
KR101519456B1	Republic of Korea	B1	2015-05-15
US9051267B2	United States of America	B2	2015-06-09
RU2014111060A	Russian Federation	A	2015-09-27
KR101563609B1	Republic of Korea	B1	2015-10-27
JP5815237B2	Japan	B2	2015-11-17
KR20150135547A	Republic of Korea	A	2015-12-02
US2016031797A1	United States of America	A1	2016-02-04
CN102702013B	China	B	2016-02-10
AU2012312902B2	Australia	B2	2016-03-17
JP5913400B2	Japan	B2	2016-04-27
EA201500845A1	Eurasian Patent Organization (EAPO)	A1	2016-04-29
CA2753436C	Canada	C	2016-05-24
EA023472B1	Eurasian Patent Organization (EAPO)	B1	2016-06-30
MX340753BThis record	Mexico	B	2016-07-25
CA2631331C	Canada	C	2016-08-09
US9409856B2	United States of America	B2	2016-08-09
US9427418B2	United States of America	B2	2016-08-30
BR112014004008A2	Brazil	A2	2017-03-28
US9624161B2	United States of America	B2	2017-04-18