Treatment methods for AR+ breast cancer

Abstract

To provide a method for treating AR+ breast cancer. The present invention provides a compound III: TIFF2023055861000019.tif47160 or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

Description

[Technical field]

[0001] Priority claim This application claims priority to U.S. Provisional Patent Application No. 62 / 353,350, filed June 22, 2016, U.S. Provisional Patent Application No. 62 / 377,497, filed August 19, 2016, and U.S. Provisional Patent Application No. 62 / 461,546, filed February 21, 2017, all of which are incorporated by reference in their entirety, including any drawings.

[0002] The present invention relates to methods of treating AR+ breast cancer. [Background technology]

[0003] background The relationship between androgens and breast cancer has been recognized for some time. In the past, androgen therapy was used to treat breast cancer in women, often with mixed success. Because breast cancer rapidly evolves to develop resistance to anti-estrogen hormone treatment, there is an urgent need to develop new treatments for breast cancer. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Gitto et al., “Recent insights into the pathophysiology of mTOR pathway dysregulation”, Res. Rep. Bio., 2:1-16 (2015) Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a method for treating AR+ breast cancer. [Means for solving the problem]

[0006] Summary of the Invention In one aspect, the present disclosure relates to a method for treating a subject (e.g., a human) with breast cancer that expresses the androgen receptor (AR+ breast cancer) by administering one or more AR agonists. In some embodiments, the AR agonist is a selective androgen receptor modulator (SARM). In some embodiments, the breast cancer is positive for both the androgen receptor and the estrogen receptor (AR+ / ER+). In some embodiments of the methods disclosed herein, the breast cancer is positive for the AR, ER, and progesterone receptor (AR+ / ER+ / PR+). In some embodiments, the AR-expressing breast cancer is not positive for Her2 (Her2 - ). In some embodiments, the breast cancer comprises one or more mutations in ER as disclosed herein. In some embodiments, the one or more ER mutations affect the ability of the ligand binding domain of ER to bind to a ligand with affinity for non-mutated ER (wild type ER). In some embodiments, breast cancers that are initially positive for ER may lose tumor expression of ER. In some embodiments, the subject is a pre- or post-menopausal woman. In some embodiments, the SARM used to treat breast cancer is used in combination with a cellular cyclin inhibitor, and in some embodiments, the cellular cyclin inhibitor is an inhibitor of CDK4 and / or CDK6 (CDK4 / 6 inhibitor). In some embodiments, the SARM is used in combination with an mTOR inhibitor. In some embodiments, the mTOR inhibitor is an inhibitor of mTOR2 and / or mTOR3. In some examples, the SARM is given by oral administration. In some embodiments, the cellular cyclin inhibitor and / or the mTOR inhibitor is given by oral administration. In some embodiments, the SARM and the mTOR inhibitor or the SARM and the CDK4 / 6 inhibitor are combined together in a kit. In some embodiments, the SARM and the mTOR inhibitor or the SARM and the CDK4 / 6 inhibitor are co-formulated.

[0007] In some embodiments, the SARM is steroidal or non-steroidal, in some embodiments, the SARM is non-steroidal.In some embodiments, the SARM of the present invention is selected from the group consisting of enobosarm, BMS-564929, LGD-4033, AC-262,356, JNJ-28330835, S-40503, LY-2452473 and GSK-2881078.In some embodiments of the methods disclosed herein, the SARM used according to the methods disclosed herein is described by the class of compounds represented by formula I and formula IV as disclosed herein.

[0008] In some embodiments of the methods disclosed herein, the cytocyclin inhibitor is a CDK4 / CDK6 inhibitor selected from the group consisting of palbociclib, ribociclib, trilaciclib, and abemaciclib. In some embodiments, the CDK4 / CDK6 inhibitor is selected from the group consisting of IC 50A compound that inhibits both CDK4 and CDK6 at <250 nM or <100 nM or <50 nM. In some embodiments of the methods disclosed herein, the mTOR inhibitor (TORC1 and / or TORC2) is selected from the group consisting of sirolimus, temsirolimus, everolimus, ridafarolimus, and MLN0128. In some embodiments, methods of treating breast cancer using a SARM in combination with a PARP inhibitor are disclosed, and in some embodiments, the PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289, ANG-3186. In some embodiments, methods of treating breast cancer using a SARM in combination with a BCL2 inhibitor are disclosed, and in some embodiments, the BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139, or S55746. In some embodiments, methods of treating breast cancer using a SARM in combination with an MCL1 inhibitor are disclosed, and in some embodiments, the MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omacataxine, seliciclib, UMI-77, AT101, sabutoclax, TW-37. In certain embodiments, methods of treating breast cancer using a SARM in combination with a PI3K inhibitor.

[0009] In some embodiments of the methods disclosed herein, the breast cancer is treatment naive. In some embodiments, the breast cancer has not yet been treated by any endocrinological treatment. In some embodiments of the methods disclosed herein, the breast cancer is resistant to at least one previous treatment. In some embodiments, the previous treatment to which resistance has occurred is at least one of antiestrogen treatment, such as an aromatase inhibitor, a selective estrogen receptor modulator (SERM) or a selective estrogen receptor degrader (SERD). In some embodiments, the subject (e.g., a woman) is postmenopausal and has progressed to a previous endocrine treatment, including but not limited to a SERD (e.g., fulvestrant, RAD1901, AZD9496); a SERM (e.g., tamoxifen, toremifene), an aromatase inhibitor, and combinations thereof. In some embodiments, the subject (e.g., a woman) has metastatic breast cancer but has not yet been treated. In some embodiments, the subject (e.g., a woman) has metastatic breast cancer and has progressed after a previous endocrinological treatment. In some embodiments, the subject (e.g., a woman) has metastatic breast cancer and has progressed after treatment with an mTOR inhibitor or a CDK4 / 6 inhibitor or a PIK3 inhibitor.

[0010] An embodiment provides a method for treating breast cancer, comprising: measuring the baseline level of ZBTB16 mRNA expression or protein expression in ER+ / AR+ breast cancer subjects; treating with AR agonist or selective androgen receptor modulator as described herein; measuring the level of ZBTB16 (encoding protein PLZF) after treatment; and if the level of ZBTB16 after treatment increases, continuing treatment with AR agonist or selective androgen receptor modulator.In an embodiment, the subject is a woman, for example a premenopausal or postmenopausal woman.

[0011] In some embodiments, an Isa diagnostic kit comprising reagents for measuring ZBTB16 mRNA or protein expression is also provided herein.

[0012] The present invention provides a method for identifying the subject who may be responsive to AR agonist treatment disclosed herein, comprising: measuring the baseline level of ZBTB16 mRNA expression or protein expression; treating with AR agonist (e.g. SARM) at a time including at least one administration; measuring ZBTB16 mRNA expression level after AR agonist treatment; and identifying the subject who may be responsive to AR agonist treatment if ZBTB16 mRNA expression increases.In some embodiments, the expression cutoff for determining responsiveness is at least x2-fold increase, x4-fold increase, x8-fold increase, x10-fold increase, x25-fold increase, x50-fold increase or >1x100-fold increase.

[0013] In some embodiments, a method is provided for treating a woman with breast cancer, wherein the woman expresses one or more mutations in estrogen receptor, such as mutations in ERα gene (ESR1). Such mutations may include fusion proteins, in which a portion of estrogen receptor is fused with a portion or all of another protein. In some embodiments, a method is provided for treating a woman with breast cancer, wherein the woman is first evaluated for one or more of the mutations and / or fusions, and if one or more mutations and / or fusions are positive, the woman is treated with an AR agonist, such as a SARM, as described herein, either as a single therapy or in combination with one or more additional chemotherapeutic agents. In some embodiments, the subject expresses mutant PI3K.

[0014] That is, the gist of the present invention relates to the following. Item 1 2. For use in a method of treating AR+ breast cancer in a subject, compound III: [ka] or a pharma- ceutically acceptable salt thereof, or a pharma- ceutically acceptable solvate thereof. Section 2 Breast cancer is AR+ / ER+ / HER2 - Item 3. The compound according to item 1, which is for breast cancer. Section 3 3. The compound according to paragraph 1 or 2, wherein the compound is administered by oral route. Section 4 (i) the subject is treated in an adjuvant setting; (ii) the subject has disease progression following treatment with one or more endocrinological agents, optionally wherein the one or more endocrinological agents are selected from the group consisting of SERMs, SERDs, progestins, aromatase inhibitors, and combinations thereof; and / or (iii) The compound of any of the preceding claims, wherein the subject has had disease progression after treatment with one or more agents selected from the group consisting of CDK4 / 6 inhibitors, mTOR inhibitors, BCL-2 inhibitors, PI3K inhibitors, and combinations thereof. Section 5 The compound of any of the preceding claims, wherein the breast cancer is resistant to at least one previous treatment. Section 6 The compound of any of the preceding claims, wherein the subject has progressed on previous endocrinological treatment. Section 7 The compound of paragraph 1 or 2, wherein the subject is a candidate for first line treatment. Section 8 7. The compound of any of the preceding claims, wherein the compound is administered at 10-500 mg, 10 mg-250 mg or 25 mg-250 mg per day, optionally said administration being once per day. Section 9 The compound of any preceding claim, wherein the subject expresses ESR1 containing one or more mutations. Section 10 10. The compound according to item 9, wherein the mutation comprises one or more mutations selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X and combinations thereof. Section 11 The compound of any preceding claim, wherein the subject expresses a PI3K mutation. Item 12 Item 12. The compound according to item 11, wherein the PI3K mutation is PIK3CA. Item 13 The compound of any of the preceding claims, wherein the treatment further comprises administration of a CDK4 / 6 inhibitor. Item 14 The compound according to item 13, wherein the CDK4 / 6 inhibitor is selected from the group consisting of palbociclib, ribociclib, trilaciclib, abemaciclib and AMG925. Item 15 The treatment further comprises a second therapeutic agent that targets the PI3K / AKT / mTOR pathway, e.g., (i) an mTOR inhibitor, optionally wherein the mTOR inhibitor is selected from the group consisting of sirolimus, temsirolimus, everolimus, ridafarolimus and MLN0128, and / or (ii) a PI3K inhibitor, optionally wherein the PI3K inhibitor is BEZ235, GDC-0980, GDC-0941, BYL719, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223; 4. The compound of any preceding claim, further comprising administration of: Item 16 The treatment, (i) PARP inhibitors, such as talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186; (ii) an MCL-1 inhibitor, optionally wherein the MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, or sabutoclax; and / or (iii) BCL-2 inhibitors, such as venetoclax, navitoclax, ABT737, G3139 or S55746; 4. The compound of any preceding claim, further comprising administration of: Section 17 The compound according to any one of items 1 to 14, wherein the treatment further comprises administration of a therapeutic agent selected from the group consisting of rapamycin, everolimus, ridaforolimus, temsirolimus, MLN0128, CC115, CC223, OSI-027, AZD8055, AZD2014, GDC-0980, SAR245409, LY3023414, NVP-BEZ235, NVP-BGT226, SF1126 and PKI-587. Section 18 Item 13. The compound according to any one of items 1 to 12, wherein the treatment further comprises administration of a therapeutic agent selected from the group consisting of everolimus, BYL719, GDC-0068, palbociclib, AZD5363 and abemaciclib, and combinations thereof. Section 19 15. The compound according to any one of items 1 to 14, wherein the treatment further comprises administration of everolimus. Section 20 15. The compound according to any one of items 1 to 14, wherein the treatment further comprises administration of BYL719. Section 21 4. The compound of any of the preceding claims, wherein said subject is a female, and optionally said female is a pre-menopausal female or a post-menopausal female. Section 22 The compound according to any of the preceding claims, wherein the breast cancer is localized, advanced or metastatic. Effect of the Invention

[0015] The present invention can provide a method for treating AR+ breast cancer. [Brief description of the drawings]

[0016] [Figure 1] Figure 1: Compound III ("RAD140") in combination with a CDK inhibitor inhibited ER+ / AR+ breast cancer growth in patient-derived xenograft (PDx) mice. [Diagram 2] Figure 2: Combined administration of RAD140 and CDK or mTOR inhibitors inhibited ER+ / AR+ breast cancer growth in cell line-derived xenograft (CDx) mice. [Diagram 3] Figure 3: RAD140 reduced ER+ / AR+ breast cancer growth in PDx mice and was more effective than fulvestrant. [Figure 4] Figure 4: RAD140 reduced ER+ / AR+ breast cancer growth in PDx mice. [Diagram 5] Figures 5A-5B: Increased ZBTB16 mRNA expression upon RAD140 treatment of breast cancer. Figure 5A: Increased ZBTB16 mRNA expression upon RAD140 treatment of T47D breast cancer cells in vitro. Figure 5B: Increased ZBTB16 mRNA expression upon RAD140 treatment of AR+ / ER+ breast cancer in vivo (PDx #2). [Figure 6] Figure 6: Inhibitory effects of RAD140, palbociclib and the combination of RAD140 and palbociclib on PDx tumors in the WHIM 18 model harboring ESR1-YAP1 fusion and E545K mutation in PIK3CA. [Figure 7] Figure 7: RAD140 inhibited the growth of ER+ / AR+ breast cancer patient-derived xenografts in the same PDX model used in Figure 3. [Figure 8] FIG. 8: RAD140 inhibited the growth of ER+ / AR+ breast cancer patient-derived xenografts in the same model used in FIG. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Detailed Description of the Invention RAD140 is an orally available, non-steroidal SARM with a distinct tissue selectivity profile. In vitro functional analysis showed that RAD140 is a potent AR agonist comparable to dihydrotestosterone in breast cancer cells. As shown in the following example section, treatment with SARM (e.g. RAD140) alone (Examples 3 and 4; Examples 7 and 8) or in combination with CDK4 / 6 inhibitors (e.g. palbociclib in Examples 1 and 2) or mTOR inhibitors (e.g. everolimus in Example 2) efficiently inhibited the growth of ER+ / AR+ breast cancer in multiple PDx and / or CDx mice. Molecular analysis of xenograft tumor biopsies revealed substantial suppression of progesterone receptor (PR), consistent with previous reports of crosstalk between AR and ER pathways (ER signaling upregulates PR), but also showed potential activation of the AR pathway in RAD140-treated tumors. Furthermore, increased ZBTB16 mRNA expression was observed upon RAD140 treatment of AR+ / ER+ breast cancer in vivo (PDx #2) and in vitro (T47D breast cancer cells), whereas PDx #2 treated with SERD (fulvestrant) showed no appreciable induction of ZBTB16 mRNA expression (Example 5). Treatment with SARM (e.g. RAD140) alone (Example 6) or in combination with CDK4 / 6 inhibitors (e.g. palbociclib) effectively inhibited the growth of ER+ / PR+ / AR+ / HER- breast cancer in WHIM18 PDx mice expressing the ESR-1 YAP fusion initially.

[0018] More specifically, RAD140 unexpectedly inhibited tumor growth in all four PDx models (PDx #1 (AR+ / ER+ / PR+ / Her2-), PDx #2 (AR+ / ER+), PDx#3 (AR+ / ER+) and WHIM18 PDx (ER+ / PR+ / AR+ / HER-)) as well as in a CDx model (ZR75 CDx (AR+ / ER+) derived from the ZR-75-1 cancer cell line (see Examples).

[0019] RAD140 alone showed tumor growth inhibition (TGI) against ER+ / PR+ / AR+ / HER- breast cancer in WHIM18 PDx, which is highly resistant to the potential ER-inhibitor Fulvestrant (Example 6). Unexpectedly, administration of RAD140 in combination with a CDK4 / 6 inhibitor (e.g., palbociclib) produced enhanced TGI effects compared to RAD140 or palbociclib treatment alone. Furthermore, in the PDx #1 (Example 1) and CDx (Example 2) models, administration of RAD140 in combination with a CDK4 / 6 inhibitor (e.g., palbociclib) again unexpectedly produced enhanced TGI effects compared to RAD140 or palbociclib treatment alone. In the CDx (Example 2) model, administration of RAD140 in combination with an mTOR inhibitor (e.g., everolimus) also produced enhanced TGI effects compared to RAD140 or palbociclib treatment alone (Example 2). Thus, SARMs (e.g., RAD140) are likely to be effective endocrine backbones to enhance the TGI of AR+ / ER+ and / or ER+ / PR+ / AR+ / HER- cancer treatment, including those that are endocrine resistant or less responsive to endocrine treatment (e.g., SERDs such as fulvestrant), and also strongly enhance the activity of active agents such as cdk4 / 6 inhibitors, m-TOR inhibitors, PI3k inhibitors, PARP inhibitors, BCL-2 inhibitors, MCL-1 inhibitors, or any combination thereof.

[0020] Based on the results provided herein, a method is provided for treating AR+ tumor in a subject in need of the treatment of AR+ tumor by administering to the subject a therapeutically effective amount of AR agonist (e.g., SARM such as RAD140), or its pharmaceutically acceptable salt or pharmaceutically acceptable solvate (e.g., hydrate).In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of one or more second therapeutic agents as described herein, such as CDK4 / 6 inhibitor, mTOR inhibitor, PARP inhibitor, PIK3 inhibitor, BCL-2 inhibitor and MCL-1 inhibitor.

[0021] The process is as follows: a) determining baseline levels of ZBTB16 mRNA and / or protein; b) administering an androgen receptor agonist; c) treating the subject with an AR agonist and then testing the level of ZBTB16 mRNA and / or protein again to provide a first level of ZBTB16 mRNA and / or protein; and d) continuing administration of the AR agonist if the first level is greater than the baseline level of ZBTB16 mRNA and / or protein. Also provided is a method for treating breast cancer in a subject, comprising:

[0022] Examples of subjects include, but are not limited to, mammals, such as humans. In some embodiments, subjects for the methods disclosed herein are women, such as premenopausal and postmenopausal women. In some embodiments, subjects for the methods disclosed herein have progressed prior to endocrine therapy, including, but not limited to, SERDs (e.g., fulvestrant, RAD1901, AZD9496); SERMs (e.g., tamoxifen, toremifene), aromatase inhibitors (e.g., arimidex, letrozole, aromasin, or combinations thereof, either directly or sequentially). In some embodiments, subjects have metastatic breast cancer and have progressed prior to endocrine therapy (e.g., SERDs, SERMs, aromatase inhibitors, or combinations thereof). In some embodiments, subjects (e.g., women) have primary or metastatic breast cancer and have progressed prior to cdk4 / 6 inhibitors, mTOR inhibitors, or PI3K inhibitors. In some embodiments, the woman is not being treated for breast cancer and an AR agonist (e.g., a SARM) (e.g., RAD140) is administered as described herein, either alone or in combination with one of the other agents described herein.

[0023] The presence of AR, ER and / or PR in breast cancer tumor cells or tumor tissues can be easily evaluated, for example, by immunohistochemistry (IHC). Some embodiments of the methods disclosed herein further comprise determining that the tumor expresses AR and optionally one or more other receptors (e.g., ER, PR and Her2), particularly ESR1. Furthermore, the methods disclosed herein develop novel treatment regimes that are not dependent on pathways that may have already been treated and become resistant (e.g., anti-estrogen resistant).

[0024] Mutations in the ERα gene (ESR1) Resistance to hormone therapy in ER+ breast cancer is often accompanied by various mutations in estrogen receptor.In some cases, these mutated receptors cause increased resistance or complete resistance to anti-estrogen / anti-endocrinological treatment, when resistance occurs as a result of treatment with aromatase inhibitors, anti-estrogen SERMs such as tamoxifen, or estrogen receptor suppressor (SERD) such as fulvestrant.In some cases, resistance to anti-estrogen treatment is accompanied by complete loss of estrogen receptor signaling due to the loss of estrogen receptor itself, but in many cases, the mutated receptor still transmits signals through the ER pathway. The reasons for anti-estrogen resistance include cases where estrogen receptors show reduced affinity for direct competitive ligands (i.e. Y537S ESR1, D538G) or mutate in such a way that they lose part or all of their ligand-binding domain (LBD) but retain enough other functional domains (e.g. DNA-binding domain, AF1 domain and / or hinge region) so that they retain constitutive activity, meaning that they remain effectively on even when they do not bind (or cannot bind) ligand.In some cases, this occurs when chromosomal translocation occurs, truncating or deleting the ligand-binding domain of ER, and another gene or partial gene is replaced at this position, resulting in a gene fusion product that produces a constitutively active receptor that does not require ligand binding.Tumor cells with such genetic changes have been shown to be resistant to therapeutic agents that target ER LBD.Such mutant cells can be enriched for a long period of time. Also, some of the mutations are known to predate treatment with anti-estrogen / SERD / aromatase inhibitors. Subjects expressing such receptors may be at higher risk of being poorly responsive to conventional anti-ER treatments, but are excellent candidates for AR agonist or SARM treatments as described herein.Thus, a patient may be a candidate for first-line or adjuvant treatment AR agonist or SARM if the subject expresses such mutation or fusion. In some embodiments, the subject is tested for the expression of candidate mutation or fusion to determine whether to treat the subject in neoadjuvant, adjuvant or first-line settings if the subject has not yet been treated with an endocrinological agent. The first-line use of SARM can be either monotherapy or combination with at least one agent selected from the group consisting of CDK inhibitors (e.g., CDK4 / 6 inhibitors), mTOR inhibitors (e.g., mTORc 1 and / or 2 inhibitors), PARP inhibitors, PIK3 inhibitors, BCL-2 inhibitors and MCL-1 inhibitors. Before the methods described herein, these mutations were particularly problematic because they prevented first-line anti-estrogenic drug treatment for ER+ breast cancer patients. As a result, patients often progress to cytotoxic agents and experience a series of their disease that is often accelerated and more difficult to treat. It is an important aspect of the present disclosure that the AR agonists described herein can effectively treat subjects with such mutant ER.

[0025] In some embodiments of the methods disclosed herein, the AR agonist disclosed herein is used to treat subjects with AR+ / ER+ breast cancer that contains one or more ER mutations. In some embodiments, these mutations affect the ability of the ligand-binding domain to bind to ligands that have affinity for non-mutated ER. In some embodiments of the present invention, the ER has one or more point mutations in the ligand-binding domain that reduce or cut off binding to ER ligands that normally bind, including agonist and / or antagonist types, including SERM and SERD, and in some cases has constitutive ER signaling activity, such as being resistant to aromatase inhibitors. In some embodiments, the mutated receptor has a ligand-binding domain that is partially or completely absent. In some embodiments, the mutated receptor is a fusion receptor between a part of ESR1 and a part or all of another protein. In some embodiments, the mutated receptor can transmit signals through the ER pathway, even though it cannot bind to ligands or has a weakened affinity for ligands that bind to non-mutated ER. In some embodiments, the mutation or fusion retains ER DNA-binding domain function.

[0026] In some embodiments of the methods disclosed herein, the subject expresses at least one of the specifically described ER fusion genes. In one embodiment, the genes include E545K mutation products in PIK3CA, ESR1-AKAP12 fusion products, ESR1-CCDC170 gene fusion products, ESR1-YAP1 gene fusion products, ESR1-POLH gene fusion products, ESR1-PCDH11X gene fusion products, or combinations thereof. In certain embodiments, the subject expresses ESR1-YAP1 fusion. In some embodiments, the subject has his / her first breast cancer or tumor that is evaluated to determine whether the subject has one or more ER gene fusion mutations in one or more samples of its cancer cells. If the subject does have one or more of the described ER mutations, the subject becomes a candidate for treatment with the compounds and methods of the present invention. In some embodiments of the present invention, the subject has already been treated with one or more previous treatments, and in some embodiments, the previous treatments include anti-estrogen treatments, such as those utilizing aromatase inhibitors, SERMs, or SERDs. In some embodiments, the subject with the indicated mutation has never been previously treated for his / her breast cancer or has never been previously treated with an antiestrogen. In some embodiments, the subject has one or more mutations / fusions that involve ligand binding function or ESR1 activity.

[0027] In some embodiments, the subject is initially positive for ER and then loses tumor expression of ER. In some embodiments, the loss of expression occurs after a series of one or more previous treatments. In some embodiments, the one or more previous treatments include treatment with an antiestrogen, further comprising one or more of an aromatase inhibitor, a SERM, and a SERD. In some embodiments, the subject who has lost ER expression in the tumor expresses the progesterone receptor (PR).

[0028] In some embodiments of the method disclosed herein, the method disclosed herein further comprises a diagnostic step, wherein the subject is first evaluated for one or more mutations / fusions as described herein.If the mutation / fusion is estimated to meet a predetermined cutoff, the subject is a candidate for either AR agonist / SARM treatment as single therapy or as combination as described herein.In some embodiments, the mutation or fusion comprises ESR1-YAP fusion and / or closely related mutations.

[0029] AR agonists The method disclosed herein is not limited to any single group of compounds, but rather includes compounds that have affinity for androgen receptor and can exhibit at least some classical androgen activity that is broadly considered to be that of AR agonist.One method for identifying such activity preclinically is in the rat Hershberger assay, where the effect of expected AR agonist is evaluated against castrated background to determine whether compound has stimulatory effect on androgen target tissues such as levator ani, prostate and / or seminal vesicle.AR agonist can be steroidal or nonsteroidal, selective or nonselective. In some embodiments, the AR agonist is a steroidal AR agonist, such as testosterone (and its esters), DHT (and its esters), fluoxymesterone, oxandrolone, stanozolol, methandrostenelone, methyltestosterone, oxymetholone, nandrolone (and its esters). In some embodiments, the AR agonist is a SARM (e.g., RAD140). In some embodiments, SARMs show efficacy against tumor endpoints, despite having a reduced androgen drive in other tissues (e.g., prostate) or other expression profiles that produce undesirable outcomes, such as virilization and hirsutism in women. In some embodiments, SARMs often have a reduced drive against liver enzyme elevations and / or against possible adverse changes in cholesterol levels, such as reduced HDL and / or increased LDL. In some embodiments, SARMs are non-steroidal and do not exhibit the potential class of liability of 17α-alkylated steroids, even though they generally still have good oral activity. In some embodiments, SARMs provide effective treatments that are less or not virilizing.In some embodiments, SARMs are less likely to feedback stimulate the central hormonal axis. In some embodiments, SARMs (e.g., RAD140) can cross the blood-brain barrier ("BBB"). In some embodiments, SARMs that can cross the BBB have an inhibitory effect on the central hormonal axis, decreasing rather than increasing sex steroids, e.g., estrogens such as estrone and estradiol, and ovarian products such as intracrine precursors such as DHEA, androstenedione. In some embodiments of the methods disclosed herein, SARMs can be used to treat premenopausal women with breast cancer and postmenopausal women with breast cancer in general. In some embodiments, inhibitory SARMs (e.g., RAD140) provide additional CNS benefits in premenopausal or postmenopausal women with breast cancer. In some embodiments, AR agonists (e.g., SARMs) disclosed herein increase lean muscle mass and / or appetite. Thus, the AR agonists (eg, SARMs) disclosed herein may be beneficial in cancer cachectic states or when wasting is a problem.

[0030] Without wishing to be bound by the examples, some of the SARMs contemplated in the methods disclosed herein include, but are not limited to, 2-chloro-4-[[(1R,2R)-2-hydroxy-2-methyl-cyclopentyl]amino]-3-methyl-benzonitrile (J Med Chem 2016; 59(2) 750), PF-06260414, enobosarm, BMS-564929, LGD-4033, AC-262356, JNJ-28330835, S-40503, GSK-2881078, RAD140, AZD-3514, MK4541, LG121071, GLPG0492, NEP28, YK11, MK0773, ACP-105, LY-2452473, S-101479, S-40542, S-42, LGD-3303, and the SARMs disclosed in US8,067,448 and US9,133,182, which are incorporated herein by reference. SARMs suitable for the methods disclosed herein also include compounds of formula I disclosed herein (e.g., Compound II and Compound III), and compounds of formula IV disclosed herein, which may be used alone or in combination with one or more agents selected from the group consisting of CDK inhibitors (e.g., CDK4 / 6 inhibitors), mTOR inhibitors (e.g., mTORc 1 inhibitors and / or mTORc 2 inhibitors), PARP inhibitors, PIK3 inhibitors, BCL-2 inhibitors, MCL-1 inhibitors, and combinations thereof, for the treatment of AR+ breast cancer in a subject.

[0031] Compounds of formula I include those of formula I: [ka] (In the formula, R x =CN; R y =CF 3 or Cl; R z =CH 3 , C.H. 2 CH 3 or Cl; or R y and R z Together: [ka] Forming; R a' is H, F, Cl, CN, OH or OSO 3 and R 1 and R 2 are each independently selected from hydrogen and methyl. The present invention includes compounds having the structure:

[0032] In some embodiments, the compound of formula I is compound II or compound III (RAD140): [ka] , a pharma- ceutically acceptable salt thereof, or a pharma- ceutically acceptable solvate thereof.

[0033] In some embodiments, the compound of formula IV: [ka] (In the formula, R x is CN, Cl, Br or NO 2 and; R y CH 3 , C.F. 3 or a halogen; R z is hydrogen, optionally substituted C 1-3 Alkyl, optionally substituted C 2-3 Alkenyl, optionally substituted C 1-3 Hydroxyalkyl, optionally substituted C 1-3 Haloalkyl, NO 2 , N.H. 2 , OMe, halogen or OH; P 1 is hydrogen or a metabolically labile group; R a and R bare each independently hydrogen or C 1-3 is alkyl; and X is O) Compounds of the formula include pharma- ceutically acceptable salts and pharma- ceutically acceptable solvates thereof.

[0034] Examples of optional substitutions include, but are not limited to, 1 to 3 halogen atoms.

[0035] In some embodiments, the SARM is a compound of formula IV, a pharma- ceutically acceptable salt, or a pharma-ceutically acceptable solvate thereof, where R x is CN.

[0036] In some embodiments, the SARM is a compound of formula IV, a pharma- ceutically acceptable salt, or a pharma-ceutically acceptable solvate thereof, where R x is CN;R y is Cl or CF 3 ;R z is hydrogen, Cl or CH 3 ;P 1 is (C=O)-C 1-6 alkyl or hydrogen; R a and R b are each independently hydrogen or -CH 3 It is.

[0037] In some embodiments, the SARM is a compound of formula IV, a pharma- ceutically acceptable salt, or a pharma-ceutically acceptable solvate thereof, where R x is CN;R y is Cl or CF 3 ;R z is hydrogen, Cl or CH 3 ;P 1 is (C=O)-C 1-6 alkyl or hydrogen; R a and R b are both hydrogen.

[0038] Combination therapy of AR agonist and mTOR inhibitor The phosphoinositide 3-kinase (PI3K) / protein kinase B (AKT) / mammalian target of rapamycin (mTOR) pathway is an intracellular signaling pathway important in regulating the cell cycle. Frequent activation of the PI3K / AKT / mTOR pathway in cancer and its critical role in cell proliferation and survival presents a challenge to find the right amount of proliferation versus differentiation to exploit this balance in the development of various therapies. See, for example, Gitto et al., "Recent insights into the pathophysiology of mTOR pathway dysregulation," Res. Rep. Bio., 2:1-16 (2015).

[0039] Inhibitors of the PI3K pathway have shown promise when offered in combination with other therapies. For example, everolimus was the first mTOR inhibitor approved in 2012 as an allosteric mTOR inhibitor, in combination with the AI ​​exemestane (Aromasin) for postmenopausal women with advanced hormone receptor-positive (HR+), HER2- breast cancer (BOLERO-2 trial). Agents targeting other components of the PI3K pathway, such as ATP-competitive dual PI3K and mTOR inhibitors (e.g., BEZ235, GDC-0980), pan-PI3K inhibitors that inhibit all four isoforms of class I PI3K (e.g., BKM120, GDC-0941), isoform-specific inhibitors of various PI3K isoforms (e.g., BYL719, GDC-0032), allosteric and catalytic inhibitors of AKT (MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363), and ATP-competitive inhibitors of mTOR only (AZD2014, MLN0128 and CC-223), are in development to treat HR+ cancers. Dienstmann et al., “Picking the point of inhibition: a comparative review of PI3K / AKT / mTOR pathway inhibitors,” Mol. Cancer Ther., 13(5):1021-31 (2014).

[0040] Despite their great potential, the undesirable side effects associated with mTOR inhibitors have hindered their development as effective cancer treatments. Kaplan et al., "Strategies for the management of adverse events associated with mTOR inhibitors", Transplant Rev (Orlando), 28(3): 126-133 (2014); and Pallet et al., "Adverse events associated with mTOR inhibitors", Expert Opin. Drug Saf. 12(2): 177-186 (2013).

[0041] Additionally, there remains a need for more durable and effective targeted therapies that can overcome the challenges associated with current endocrine therapies, while combining with second therapeutic agents (e.g., everolimus and other agents targeting the PI3K / AKT / mTOR pathway) may provide additional benefit in advanced stages and / or against cancers resistant to previous treatments.

[0042] In some embodiments, the second therapeutic agent targets the PI3K / AKT / mTOR pathway and can be an mTOR inhibitor, a dual mTOR inhibitor, a PI3K / mTOR inhibitor, or an inhibitor of mTOR2 and / or mTOR3. In some embodiments, the second therapeutic agent is a rapamycin derivative (known as a rapalog), such as rapamycin (sirolimus or Rapamune, Pfizer), everolimus (Affinitor or RAD001, Novartis), ridaforolimus (AP23573 or MK-8669, Merck and ARIAD Pharmaceuticals), temsirolimus (Torisel or CCI779, Pfizer), including solvates (e.g., hydrates) and salts thereof. In some embodiments, the second therapeutic agent is a dual mTOR inhibitor that inhibits both mTORC1 and mTORC2, such as MLN0128, CC115 and CC223 (Celgene), OSI-027 (OSI Pharmaceuticals), and AZD8055 and AZD2014 (AstraZeneca), including solvates (e.g., hydrates) and salts thereof. In some embodiments, the second therapeutic agent is a PI3K / mTOR inhibitor, such as GDC-0980, SAR245409 (XL765), LY3023414 (Eli Lilly), NVP-BEZ235 (Novartis), NVP-BGT226 (Novartis), SF1126 and PKI-587 (Pfizer), including solvates (e.g., hydrates) and salts thereof.

[0043] In some embodiments, more than one of the above-mentioned second therapeutic agents can be used in combination with the AR agonist (e.g., SARM) disclosed herein, such as the compound of formula I, the compound of formula IV, compound II and compound III.For example, an mTOR inhibitor can be used together with another mTOR inhibitor or a PI3K / mTOR inhibitor.It is also known in the art that the above-mentioned second therapeutic agents, including mTOR inhibitors, dual mTOR inhibitors and PI3K / mTOR inhibitors, can be administered together with other active agents to enhance the effectiveness of treatment, for example, JAK2 inhibitors (Bogani et al., PLOS One, 8(1): e54826 (2013)), chemotherapeutic agents (Yardley, Breast Cancer (Auckl) 7: 7-22 (2013)).Therefore, the second therapeutic agent also includes these supplementary active agents.

[0044] FIG. 2 shows the increased efficacy obtained when the mTOR inhibitor everolimus was used in combination with the SARM RAD140 in an in vivo model.

[0045] Combination therapy of AR agonist and CDK inhibitor Cell cycle regulators such as cyclins and cyclin-dependent kinases (CDKs) have been reported to have effects on ER expression. Lamb et al., "Cell cycle regulators cyclin D1 and CDK4 / 6 have estrogen receptor-dependent divergent functions in breast cancer migration and stem cell-like activity", Cell Cycle 12(15): 2384-2394 (2013). Selective CDK4 / 6 inhibitors (e.g., ribociclib, abemaciclib, and palbociclib) have targeted tumor types in which CDK4 / 6 plays a central role in the G1- to S-phase cell cycle transition with improved efficacy and reduced deleterious effects on normal cells. O'Leary et al., "Treating cancer with selective CDK4 / 6 inhibitors," Nat. Rev. Clin. Oncol. (2016), published March 31, 2016 (http: / / www.nature.com / nrclinonc / journal / vaop / ncurrent / full / nrclinonc.2016.26.html). Selective CDK4 / 6 inhibitors showed the best response when tested in combination with ER endocrine therapy in patients with ER-positive breast cancer. In this regard, it is useful to consider that the AR pathway may interact with the ER pathway to antagonize ER activity in a manner that differs from direct ER antagonism under some conditions.

[0046] Palbociclib in combination with the aromatase inhibitor, letrozole (PALoMA-1 / TRIO 18 study) was approved in February 2015 as the first endocrine-based therapy in postmenopausal women for the treatment of hormone receptor (HR)-positive (HR+), HER2-negative (HER2-) advanced breast cancer. In February 2016, palbociclib in combination with the SERD fulvestrant (PALOMA-3 study) was approved for the treatment of patients with ER+, HER2- advanced or metastatic breast cancer who had progressed prior to endocrine therapy. The FDA granted breakthrough therapy designation to the CDK4 / 6 inhibitor abemaciclib (LY2835219) as monotherapy for severely pretreated patients with refractory HR-positive advanced breast cancer based on data from a phase I study (JPBA study). Further combinations of selective CDK4 / 6 inhibitors (e.g., ribociclib, abemaciclib, and palbociclib) with ER endocrine therapies (e.g., AIs, SERMs, and SERDs) are currently under development. However, to date there appears to be a lack of clinical or preclinical studies demonstrating the efficacy of unexpected combinations between SARMs and cdk4 / 6 inhibitors.

[0047] Moreover, CDK4 / 6 inhibitors show toxicity that may require cyclical treatment (O'Leary).Therefore, there remains a need for effective and alternative combination-targeting treatments that can solve the problems associated with current endocrine treatments and provide additional benefits by combining with CDK4 / 6 inhibitors against breast cancer, especially breast cancers that are in advanced stages and / or have resistance to previous treatments or have mutations identified as existing before or after previous treatments that make ESR1 target responsive to androgen intervention (including SARMs), where other endocrinological treatments, especially those that rely on direct anti-estrogenic effects such as aromatase inhibitors, SERMs or SERDs, are generally less effective or not effective at all.

[0048] In some embodiments, CDK4 and / or CDK6 inhibitors include, but are not limited to, palbociclib, abemaciclib, ribociclib, and AMG925.

[0049] An in vivo illustration of this concept of an AR agonist (e.g., SARM) in combination with a CDK4 / CDK6 inhibitor (e.g., palbociclib) can be seen in FIG. 1. As shown in FIG. 1, the SARM RAD140 has a highly effective tumor suppression ability similar to monotherapy with palbociclib. It was unexpected that the combination treatment of RAD140 and palbociclib provided enhanced TGI compared to monotherapy with RAD140 or palbociclib. Thus, the combination treatment of an AR agonist and a CDK4 / 6 inhibitor may provide improved activity and significant clinical benefits for the treatment of AR+ breast cancer. Similarly, FIG. 6 shows the efficacy of either alone or together with palbociclib in the WHIM18 model expressing the ESR1-YAP fusion for the first time, where fulvestrant (as a SERD) was not effective alone and did not enhance the efficacy of palbociclib in the fulvestrant-palbociclib combination treatment.

[0050] Administration and Formulation With regard to the administration of the compounds and combinations according to the methods disclosed herein, the AR agonist (e.g., SARM) disclosed herein or its solvate or salt and CDK4 and / or CDK6 inhibitor (e.g., ribociclib, abemaciclib and palbociclib) or mTOR inhibitor, PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor and / or BCL2 inhibitor are administered in combination to a subject in need thereof. The phrase "in combination" means that the AR agonist (e.g., SARM) disclosed herein can be administered before, during or after the administration of CDK4 and / or CDK6 inhibitor or mTOR inhibitor, PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor and / or BCL2 inhibitor. For example, the AR agonists (e.g., SARMs) and CDK4 and / or CDK6 inhibitors or mTOR inhibitors, PI3K inhibitors, PARP inhibitors, MCL-1 inhibitors, and / or BCL2 inhibitors can be administered at about 1 week apart, about 6 days apart, about 5 days apart, about 4 days apart, about 3 days apart, about 2 days apart, about 24 hours apart, about 23 hours apart, about 22 hours apart, about 21 hours apart, about 20 hours apart, about 19 hours apart, about 18 hours apart, about 17 hours apart, about 16 hours apart, about The administration may be performed at about 15 hours apart, about 14 hours apart, about 13 hours apart, about 12 hours apart, about 11 hours apart, about 10 hours apart, about 9 hours apart, about 8 hours apart, about 7 hours apart, about 6 hours apart, about 5 hours apart, about 4 hours apart, about 3 hours apart, about 2 hours apart, about 1 hour apart, about 55 hours apart, about 50 hours apart, about 45 hours apart, about 40 hours apart, about 35 hours apart, about 30 hours apart, about 25 hours apart, about 20 hours apart, about 15 hours apart, about 10 hours apart, or about 5 hours apart. In some embodiments, the AR agonist (e.g., SARMs) and the CDK4 and / or CDK6 inhibitor or mTOR inhibitor, PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor, and / or BCL2 inhibitor are administered to the subject simultaneously or substantially simultaneously.In some of these embodiments, the AR agonist (e.g., SARMs) disclosed herein and CDK4 and / or CDK6 inhibitor (e.g., ribociclib, abemaciclib and palbociclib) or mTOR inhibitor (e.g., sirolimus, temsirolimus, everolimus, ridafalolimus and MLN0128), PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor and / or BCL2 inhibitor can be administered as part of a single formulation.Included are kits in which the AR agonist and one or more additional agents described herein are included together in a kit, for example, as a co-packaged arrangement.As a non-limiting example, the scope includes a kit that includes RAD140 and a CDK4 / 6 inhibitor, an m-TOR inhibitor, a PI3K inhibitor, a PARP inhibitor, an MCL-1 inhibitor and / or a BCL2 inhibitor, for example, as detailed herein.

[0051] In some embodiments, a single AR agonist (e.g., SARM) and a single CDK4 and / or CDK6 inhibitor or a combination of mTOR inhibitor, PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor and / or BCL2 inhibitor are administered to a subject.In some embodiments, a combination of an AR agonist (e.g., SARM) and a CDK4 / CDK6 inhibitor and an mTOR inhibitor, PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor and / or BCL2 inhibitor are administered to a subject together.The formulation of AR agonist (e.g., SARM) used in the method disclosed herein is generally disclosed in the literature, and these teachings are incorporated herein by general reference.In particular, US8,067,448 discloses how to make the compound, and the general formulation method useful for formulating the compound of formula I, compound II and compound III is incorporated herein by reference. Similarly, US9,133,182 discloses how to make and generally formulate compounds of formula IV, and is incorporated herein by reference. In instances where specific preparation advice or instructions are not available, some general principles of formulation may apply. For example, the compounds and combinations of the methods disclosed herein may be formulated into unit dosage forms, which refer to physically separate units suitable as unitary doses for the subject undergoing treatment, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect, optionally with a suitable pharmaceutical carrier. The unit dosage form may be for a single daily dose or may be one of multiple daily doses (e.g., about 1-4 or more qd). When multiple daily doses are used, the unit dosage form may be the same or different for each dose. In some embodiments, the compounds may be formulated for controlled release.

[0052] The compound and combination for use in the method disclosed in the present invention can be formulated according to any available conventional method.Examples of preferred dosage forms include tablets, powders, subtle granules, granules, coated tablets, capsules, syrups, troches, inhalants, suppositories, injections, ointments, eye ointments, eye drops, nose drops, ear drops, poultices, lotions, etc. In the preparation, commonly used additives such as diluents, binders, disintegrants, lubricants, colorants, flavorings, and, if necessary, stabilizers, emulsifiers, absorption promoters, surfactants, pH adjusters, preservatives, antioxidants, etc. can be used.In addition, formulation can be carried out by combining compositions commonly used as raw materials for pharmaceutical preparations according to conventional methods. Examples of these compositions include, for example, (1) oils such as soybean oil, beef tallow, and synthetic glycerides; (2) hydrocarbons such as liquid paraffin, squalane, and solid paraffin; (3) ester oils such as octyldodecyl myristate and isopropyl myristate; (4) higher alcohols such as cetostearyl alcohol and behenyl alcohol; (5) silicone resins; (6) silicone oils; (7) polyethylene fatty acid esters, sorbitan fatty acid esters, glycerin fatty acid esters, polyoxyethylene sorbitan fatty acid esters, solid polyoxyethylene castor oil, and polyoxyethylene polyoxypropylene. (8) water-soluble macromolecules such as hydroxyethyl cellulose, polyacrylic acid, carboxyvinyl polymers, polyethylene glycol, polyvinylpyrrolidone, and methylcellulose; (9) lower alcohols such as ethanol and isopropanol; (10) polyhydric alcohols such as glycerol, propylene glycol, dipropylene glycol, and sorbitol; (11) sugars such as glucose and sucrose; (12) inorganic powders such as silicic anhydride, aluminum magnesium silicate, and aluminum silicate; and (13) purified water.Excipients for use in the above formulations include, for example: 1) diluents such as lactose, corn starch, sucrose, glucose, mannitol, sorbitol, crystalline cellulose, and silicon dioxide; 2) binders such as polyvinyl alcohol, polyvinyl ether, methylcellulose, ethylcellulose, gum arabic, tragacanth, gelatin, shellac, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polypropylene glycol-polyoxyethylene-block copolymer, meglumine, calcium citrate, dextrin, pectin, and the like; and 3) disintegrants. Examples of suitable additives include starch, agar, gelatin powder, crystalline cellulose, calcium carbonate, sodium bicarbonate, calcium citrate, dextrin, pectic, carboxymethylcellulose / calcium, etc.; 4) lubricants include magnesium stearate, talc, polyethylene glycol, silica, concentrated vegetable oils, etc.; 5) colorants include any colorant whose addition is pharma- ceutically acceptable and appropriate; 6) flavoring agents include cocoa powder, menthol, aromatizing agents, peppermint oil, cinnamon powder; and 7) antioxidants such as ascorbic acid or alpha-tophenol whose addition is pharma- ceutically acceptable.

[0053] The compounds and combinations for use in the methods disclosed in the present invention can be formulated into pharmaceutical compositions as any one or more of the active compounds described herein and physiologically acceptable carriers (also referred to as pharmaceutically acceptable carriers or solutions or diluents).Such carriers and solutions include the pharmaceutically acceptable salts and solvates of the compounds used in the methods of the present invention, as well as mixtures that contain two or more of such compounds, pharmaceutically acceptable salts of the compounds, and pharmaceutically acceptable solvates of the compounds.Such compositions are prepared according to acceptable pharmaceutical procedures, such as those described in Remington's Pharmaceutical Sciences, 17th edition, edited by Alfonso R. Gennaro, Mack Publishing Company, Eaton, Pa. (1985), which is incorporated herein by reference.

[0054] The term "pharmaceutically acceptable carrier" refers to a carrier that is compatible with other ingredients in the formulation without causing allergic reactions or other untoward effects in the patient to which it is administered. Pharmaceutically acceptable carriers include, for example, pharmaceutical diluents, excipients or carriers that are appropriately selected with respect to the intended form of administration and are not inconsistent with conventional pharmaceutical practice. For example, solid carriers / diluents include, but are not limited to, gums, starches (e.g., corn starch, pregelatinized starch), sugars (e.g., lactose, mannitol, sucrose, dextrose), cellulose materials (e.g., microcrystalline cellulose), acrylates (e.g., polymethyl acrylate), calcium carbonate, magnesium oxide, talc or mixtures thereof. Pharmaceutically acceptable carriers may further include minor amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the therapeutic agent.

[0055] The free form of the AR agonist (e.g., SARM) and / or CDK4 / 6 inhibitor and / or mTOR inhibitor, PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor and / or BCL2 inhibitor may be converted to a salt, if desired, by conventional methods. The term "salt" as used herein is not limited as long as the salt is pharma- ceutically acceptable, and preferred examples of salts include hydrohalide salts (e.g., hydrogen chloride, hydrogen bromide, hydrogen iodide, etc.), inorganic acid salts (e.g., sulfate, nitrate, perchlorate, phosphate, carbonate, bicarbonate, etc.), organic carboxylate salts (e.g., acetate, maleate, tartrate, fumarate, citrate, etc.), organic sulfonate salts (e.g., methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, camphorsulfonate, etc.), amino acid salts (e.g., aspartate, glutamate, etc.), quaternary ammonium salts, alkali metal salts (e.g., sodium salt, potassium salt, etc.), alkaline earth metal salts (magnesium salt, calcium salt, etc.), etc. Hydrochloride salts, sulfate salts, methanesulfonate salts, acetate salts, etc. are preferred as "pharmaceutical acceptable salts" of the compounds disclosed herein.

[0056] In some embodiments, the AR agonists (e.g., SARMs) and CDK4 / 6 and / or mTOR inhibitors, PI3K inhibitors, PARP inhibitors, MCL-1 inhibitors and / or BCL2 inhibitors disclosed herein may be in a prodrug form, meaning that they must undergo some transformation (e.g., oxidation or hydrolysis) to achieve their active form.

[0057] Administration of the compounds and / or combinations disclosed herein can be by routes previously described for these compounds, but generally can be transdermal, subcutaneous, intravenous, intranasal, pulmonary, oral, etc. For the combination methods of the present invention, oral is the preferred route.

[0058] The therapeutically effective amount of the combination of AR agonist (e.g., SARM) and CDK4 / 6 inhibitor and / or mTOR inhibitor, PI3K inhibitor, PARP inhibitor, MCL-1 inhibitor and / or BCL2 inhibitor in the method disclosed herein is an amount that, when administered over a certain time interval, results in the achievement of one or more therapeutic levels (e.g., causing tumor growth to be delayed or stopped, tumor regression, symptoms to be stopped, etc.). The combination for use in the method disclosed in the present invention can be administered to a subject once or multiple times. In those embodiments in which the compound is administered multiple times, the compound can be administered at set intervals, such as daily, every other day, weekly, or monthly. Alternatively, the compound can be administered at irregular intervals, such as on a required basis based on symptoms, the patient's health, etc. The therapeutically effective amount of the combination can be administered qd for 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 10 days, or at least 15 days. Optionally, the cancer status or tumor regression is monitored during or after treatment, for example by FES-PET scanning of the subject. The dose of the combination administered to the subject can be increased or decreased depending on the cancer status or tumor regression detected.

[0059] Those skilled in the art can easily determine this amount either on an individual subject basis (e.g., the amount of compound required to achieve a particular therapeutic level in the subject being treated) or on a population basis (e.g., the amount of compound required to achieve a particular therapeutic level in the average subject of a given population). Ideally, a therapeutically effective amount does not exceed the maximum tolerated dose at which 50% or more of treated subjects experience nausea, hirsutism, hoarseness, or other more severe reactions that prevent further drug administration. A therapeutically effective amount may vary for a subject depending on a variety of factors, including the change and degree of symptoms, the sex, age, weight, or general health of the subject, the dosage form and salt or solvate type, the variation in sensitivity to the drug, the particular type of disease, and the like. One means of indicating an acute response to the treatment regimen of the present invention is to analyze progestin receptor expression. It has been found that the AR agonists (e.g., SARMs) used in the methods of the present invention cause a decrease in the expression of progestin receptors, which indicates a response to the drug. Based on the extensive preclinical efficacy data in mouse xenografts presented in the Examples, calculation and disclosure of a predicted effective human clinical dose for the SARM RAD140 is provided in Example 9.

[0060] The method examples disclosed herein are for illustrative purposes and the invention is not limited to the embodiments exemplified. EXAMPLES

[0061] Working Example Example 1: SARM RAD140 in combination with a CDK inhibitor inhibited the growth of ER- and AR-positive breast cancer in PDx mice (Figure 1) material and method: PDx model #1, maintained by a Contract Research Organization (CRO), was characterized as AR+ / ER+ / PR+ / Her2- using IHC and gene chip microarrays. Donor tumor slices were implanted subcutaneously into the flank of intact female nude mice (n=7). 3Animals bearing tumors measuring 2,000 mm in size were randomized into four treatment groups. Animals in each group received vehicle (circles, FIG. 1), RAD140 100 mg / kg twice daily (bid) (triangles, FIG. 1), palbociclib 75 mg / kg once daily (qd) (squares, FIG. 1), or a combination of RAD140 100 mg / kg (bid) and palbociclib 75 mg / kg (qd) (diamonds, FIG. 1). All test compounds were administered orally for 42 consecutive days. Tumor volumes were measured twice weekly and % tumor growth inhibition (TGI) was calculated. Tumor volumes were measured twice weekly and % tumor growth inhibition (TGI) was calculated. 3 Animals with tumors larger than 1 mm in size were euthanized in accordance with animal welfare regulations. At the end of the study, plasma and tumor samples were collected for pharmacokinetic and pharmacodynamic analysis. To stimulate tumor growth, mice were supplemented with estradiol in the water.

[0062] result: Treatment with RAD140 or palbociclib alone resulted in inhibition of tumor growth with approximately 53% TGI (tumor growth inhibition), respectively (Figure 1). Combined administration of SARM and CDK4 / 6 inhibitor resulted in 70% TGI at day 28, at which point the number of animals in the vehicle-treated group was less than six due to ethical handling of mice with larger tumors. The combination of RAD140 and palbociclib continued to exert potent tumor-suppressing effects until the end of the study (day 42). The estimated end-point TGI was greater than 70%. No appreciable RAD140 treatment-related weight loss was observed (data not shown).

[0063] In summary, treatment with SARMs alone showed comparable tumor inhibition to treatment with palbociclib in AR+ / ER+ / PR+ / Her2- breast cancer PDx-bearing mice. Combined administration of RAD140 and palbociclib produced greater growth inhibition effects in these xenografts than RAD140 and palbociclib acting alone, respectively. These results indicate that combined administration of SARMs with or without CDK4 / 6 inhibitors is effective in ER+ / AR+ breast cancer, and that combination with CDK4 / 6 enhanced TGI more than SARMs and CKD4 / 6 inhibitors could do when administered alone, respectively.

[0064] Example 2: Administration of the SARM RAD140 in combination with a CDK inhibitor or an mTOR inhibitor inhibited ER- and AR-positive breast cancer growth in CDx mice (Figure 2) material and method: ZR-75-1 is a frequently used breast cancer cell line model that is ER+ / AR+. ZR-75-1 cells (donor tumors) were injected into the flanks of female nude mice to establish the parental ZR-75-1 CDx model (ZR75 model). The ER+ / AR+ status of these donor tumors was confirmed using immunoblotting (IB) and IHC. Donor tumor sections were implanted subcutaneously into the flanks of intact female nude mice (n=7). After randomization of established xenograft tumors, animals in each group received vehicle (diamond, Figure 2), RAD140 100 mg / kg (bid) (square, Figure 2), palbociclib 45 mg / kg (qd) (triangle, Figure 2), a combination of RAD140 100 mg / kg (bid) and palbociclib 45 mg / kg (qd) (star, Figure 2), everolimus 2.5 mg / kg (qd) ("X", Figure 2), or a combination of RAD140 100 mg / kg (bid) and everolimus 2.5 mg / kg (qd) (circle, Figure 2). All test compounds were administered orally for 28 consecutive days. ZR75-1 xenograft tumors in animals were supported with estradiol pellets (0.18 mg 90-day release, Innovative Research America). Tumor volumes were measured twice a week and %TGI was calculated. 3 Animals bearing tumors exceeding a size of 100 were euthanized in accordance with animal welfare regulations. At the end of the study, plasma and tumor samples were collected for pharmacokinetic and pharmacodynamic analysis.

[0065] result: Treatment with RAD140, palbociclib or everolimus alone resulted in inhibition of tumor growth with TGI of 52%, 84% or 75%, respectively (Figure 2). Combinations of RAD140 with palbociclib or everolimus showed potent tumor suppressive effects until the end of the study (day 28). End point TGI for the RAD140-palbociclib and RAD140-everolimus combinations were 96% and 101%, respectively. No appreciable RAD140 treatment-related weight loss was observed (data not shown).

[0066] In summary, treatment with RAD140 alone showed antitumor activity in ER+ / AR+ breast cancer CDx mice. Combined administration of RAD140 with either palbociclib or everolimus produced enhanced growth inhibitory effects in these xenografts compared with RAD140, palbociclib, or everolimus alone, respectively. These results indicate that combined administration of the SARM RAD140 alone or in combination with either a CDK4 / 6 inhibitor or mTOR is effective in ER+ / AR+ breast tumors.

[0067] Example 3: SARMs reduced ER and AR breast cancer growth in PDx models and were more effective than fulvestrant (Figure 3) material and method: CRO-maintained PDx model #2 was characterized as AR+ / ER+ using IHC and gene chip microarray. Donor tumor sections were implanted subcutaneously into the flank of intact female nude mice (n=10). After randomization of established xenografts, animals in each group received vehicle (diamond, Figure 3), RAD140 100 mg / kg daily (qd) (square, Figure 3), fulvestrant 1 mg weekly (qw) (circle, Figure 3) or a combination of RAD140 100 mg / kg (qd) and fulvestrant 1 mg (qw) (star, Figure 3). RAD140 was administered orally for 42 consecutive days and fulvestrant was administered subcutaneously every other week for 6 weeks. Tumor volumes were measured twice weekly to calculate %TGI. 2,000 mm 3 Animals with tumors larger than 1 mm in size were euthanized in accordance with animal welfare regulations. The PDx model was supplemented with estradiol in the water to stimulate tumor growth. At the end of the study, plasma and tumor samples were collected for pharmacokinetic and pharmacodynamic analysis.

[0068] result: Treatment with RAD140 alone resulted in inhibition of tumor growth with approximately 76% TGI (Figure 3). Fulvestrant alone resulted in 59% TGI. Combined administration of RAD140 and fulvestrant resulted in a 76% TGI similar to that observed with RAD140 alone. No appreciable SARM treatment-related weight loss was observed (data not shown).

[0069] In summary, SARMs alone demonstrated more effective antitumor activity than SERDs (e.g., fulvestrant, a standard of care drug for ER+ breast cancer). Combined administration of SARMs with fulvestrant did not demonstrate improved efficacy over that of RAD140 alone in ER+ / AR+ breast cancer PDx mice.

[0070] Example 4: RAD140 reduced ER- and AR-positive breast cancer growth in a PDx model (Figure 4) material and method: CRO-maintained PDx model #3 was characterized as AR+ / ER+ using IHC and gene chip microarray. Donor tumor sections were implanted subcutaneously into the flank of intact female nude mice (n=10). After randomization of established xenografts, animals in each group received vehicle (diamonds, FIG. 4) or RAD140 100 mg / kg (bid) (triangles, FIG. 4) for 45 days. PDx mice were supplemented with estradiol in water to stimulate tumor growth.

[0071] result: Treatment with RAD140 resulted in inhibition of tumor growth with a TGI of 49% (Figure 4). No appreciable RAD140 treatment-associated weight loss was observed (data not shown).

[0072] In summary, these results demonstrate that SARMs inhibit the growth of ER+ / AR+ breast tumors.

[0073] Example 5: RAD140 induces AR target gene ZBTB16 expression in ER+ / AR+ breast cancer cells and patient-derived xenografts (PDx) (Figure 5) material and method: PDX model #2 was treated as described in Example 3, and tumor samples were frozen after 6 hours of the last dose. ER and AR positive T47D breast cancer cells were incubated in medium supplemented with 5% charcoal-dextran stripped serum (CSS) for 48 hours and then treated with vehicle (DMSO), 1nM, 10nM, 100nM, 1,000nM RAD140, or 1nM or 10nM DHT. Cells were harvested 24 hours after treatment. RNA was extracted from frozen tumor samples from the above PDX #2 and T47D cells using Qiagen RNeasy kit. Real-time quantitative PCR (qPCR) was performed using primer / probe sets (Applied Biosystems / ThermoScientific) for AR target genes ZBTB16 (encoding PLZF protein) and GAPDH (internal control).

[0074] result: Treatment of T47D cells with RAD140 in vitro resulted in a dose-dependent increase in ZBTB16 mRNA expression (Figure 5A), with 1,000 nM treatment resulting in an approximately 130-fold increase compared to vehicle-treated cells. The natural androgen DHT also resulted in a 300-500-fold induction of this gene. This further supports that RAD140 is an effective AR agonist. Consistently, approximately 25-fold induction of the ZBTB16 gene was also seen in PDX #2 treated with RAD140 100 mg / kg (mpk) qd for 45 days compared to that of vehicle-treated tumors (Figure 5B). In contrast, fulvestrant, an antagonist of the SERD and ER pathways, did not result in an appreciable increase in ZBTB16 gene expression (Figure 5B). ZBTB16 / PLZF has been implicated in prostate cancer as a tumor suppressor that is suppressed in recurrent tumors after androgen deprivation treatment (ADT, also known as castration). These results suggest that the SARM RAD140 activated the transcription of AR target genes in breast cancer cells. More importantly, RAD140 suppressed breast cancer growth by inducing certain tumor suppressor genes, including but not limited to ZBTB16 / PLZF.

[0075] Example 6: RAD140 was effective in the WHIM18 PDx model with and without palbociclib The WHIM18 PDx model is a xenograft of patient-derived breast cancer tumors implanted in thymic ablated female nude mice. WHIM18 xenografts are ER+ / PR+ / AR+ / HER- and grew independently of exogenous estrogen (e.g., 17β-estradiol). The WHIM18 PDx model was highly resistant to the potent ER inhibitor fulvestrant. The WHIM18 PDx model harbored an ESR1-YAP1 fusion and an E545K mutation in PIK3CA.

[0076] The efficacy of RAD140 or fulvestrant alone and in combination with palbociclib in the WHIM18 PDx model was evaluated over 56 days (8 weeks) of treatment. The primary endpoint was tumor growth. EDTA plasma and tumor tissues were collected after the last dose.

[0077] material and method Female outbred thymic nude mice (The Jackson Laboratory 007850) (DOB 7-19-16) were inoculated with 1.5x10 6 A single cell suspension of WHIM18 cells, passage 9, was implanted. The cells were mixed 1:1 with DMEM:Matrigel (Corning REF 354234) in a total volume of 100 μl / mouse. 3 Sixty mice were analyzed by tumor volume using Biopticon's Tumor Manager 62 days after cell implantation when the tumor reached an average volume of 179.9 mm (actual mean = 179.9 mm). TM Software was used to randomize into one of six treatment groups (10 mice / group).

[0078] Each mouse was dosed for 56 days with the dosing schedule described in Table 1, receiving vehicle 0.5% carboxymethylcellulose (CMC, Sigma C4888), RAD140 or palbociclib by oral gavage once daily and fulvestrant by subcutaneous injection once every 7 days. Dose volumes were calculated based on the average weekly animal weight for each group shown in Table 1. 0.5% CMC was prepared by dissolving 2.5 g in 400 ml of sterile warm water and heating with stirring until dissolved, qs up to 500 ml with sterile water and stored at 4°C. [Table 1]

[0079] The RAD140 compositions for administration in this example were prepared by adding an appropriate amount of 0.5% CMC to RAD140 with continuous stirring, protected from light, at 4° C. The RAD140 compositions were prepared weekly.

[0080] The palbociclib compositions for administration in this example were prepared by adding an appropriate amount of sterile saline (0.9% sodium chloride for injection, NDC 0409-7983-03) to palbociclib with stirring. The palbociclib compositions were stirred continuously at 4° C. until the palbociclib was dissolved. The palbociclib compositions were prepared weekly.

[0081] Biopticon's Tumor Imager TM Tumor volumes were measured twice a week using the corresponding TumorManager TM The volume was calculated using software. Mice were weighed once a week during the first half of the study (day 27). Once mice showed weight loss, they were weighed twice a week. Additionally, any mice that had a body weight loss (BWL) of ≥ 5% compared to day 0 or showed significant clinical signs (e.g., hunched posture, dirty appearance) were weighed daily. Treatment was discontinued for any mice that had a BWL of ≥ 15% compared to day 0, and treatment was resumed if the mice's weight returned to ≤ 15% BWL.

[0082] When the mouse becomes moribund, the tumor volume of the mouse is 2,000 mm 3 Mice were removed from the study if the time to administration of the drug was exceeded or if the mice lost > 20% of their body weight compared to day 0. When possible, test samples were collected and the time of last administration and take down were recorded.

[0083] The remaining animals were taken down 6 hours after the last dose on day 56. The last dose of fulvestrant was given on the morning of sacrifice. Actual sacrifice took place between 6 and 9 hours. 2 Immediately after death, blood was collected by cardiac puncture and spun down in EDTA tubes at 2000 g for 10 min at 4°C. Plasma was transferred to Eppendorf tubes and stored at -80°C.

[0084] After blood collection, tumors were excised and weighed. For each tumor, one half of the tumor was placed in 10% neutral buffered formalin (NBF) and the other half of the tumor was placed in an Eppendorf tube and diluted with liquid N 2 The tissues were then flash frozen in PBS and stored at -80°C. All tissues retrieved for formalin-fixed, paraffin-embedded (FFPE) blocks were fixed in neutral buffered formalin (NBF) for 24–48 hours and then transferred to 70% ethanol for subsequent shipping to be processed into FFPE blocks.

[0085] result: The results of the in vivo experiments are shown graphically in Figure 6. The relative reduction in tumor growth is statistically shown in Tables 2 and 3 below.

[0086] Tumor growth inhibition (TGI, %) was calculated relative to the vehicle group on day 56 (Table 2, formula 1).

number

[0087] Student's t-test was calculated in Excel using two-way distribution of Δtumor volume day 56-day 0 and equal variance of the two samples. [Table 3]

[0088] Administration of RAD140 alone or in combination with palbociclib at a dose of 100 mg / kg inhibited the growth of HER2-, ER+, PR+ breast cancer tumors implanted in female thymic nude mice (WHIM18 PDx) (Figure 6). The combination of RAD140 and palbociclib appears to be more effective than either drug alone. No obvious toxicity was observed in either group.

[0089] Example 7 In PDx model #2, the same PDx model as described in the Examples / FIG. 3, RAD140 administered at low doses of both 1 mg / kg bid or 10 mg / kg bid resulted in substantial inhibition of tumor growth as judged by TGI values ​​of approximately 81% in both groups.

[0090] Example 8 In PDx model #1, the same PDx model as described in the Examples / FIG. 1, RAD140 administered at low doses of 1 mg / kg bid, 3 mg / kg bid or 10 mg / kg bid all resulted in substantial inhibition of tumor growth as judged by TGI values ​​of 49%, 65% and 57%, respectively.

[0091] Example 9 RAD140 was shown to have good activity in PDx tumor xenografts implanted in nude mice. Activity was significant in the range of 1 mg / kg to 100 mg / kg. Considering the specific exposure levels in mice, and cross species pharmacokinetic modeling from both known and derived pharmacokinetic parameters, a dose range in female patients can be calculated. Notably, doses from 1 mg / kg (bid) to 100 mg / kg (bid) were all shown to have good efficacy in one or more of the models described in the Examples herein. Based on the interspecies half-life and microsomal stability, RAD140 is predicted to be effective as a once-daily oral dose in the dose range of 5 mg to 500 mg. For example, a mouse effective dose of 10 mg / kg (qd) would be effectively translated to a dose of approximately 50 mg qd in a 60 kg woman. A range of 1 mg / kg to 100 mg / kg (bid) has been shown to be effective, so a broader range of 10 mg to 1000 mg is clinically evident. Particularly within this range, further ranges of 10 mg to 250 mg, 25 mg to 250 mg, 25 to 500 may also be found to be supported. Similarly, specific points within the range, integer or non-integer, are supported, as are individual dose points that fall within any of the ranges. For example, doses such as 12.5 mg, 17.5 mg, etc. are specifically contemplated, as are doses such as 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, and 500 mg. Further non-limiting examples include doses of 10 mg, 15 mg, 20 mg, 25 mg, 50 mg, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500 mg. QD administration of the doses described is expected to be highly suitable as pharmacokinetic studies in animals predict that RAD140 has a long half-life suitable for once-daily administration, although bid administration also works, however the doses listed above for once-daily administration are split into two doses so that they are given twice a day.

[0092] Example 10: Method 1: administering to a subject a dose of Formula I [ka] (In the formula, R x =CN; R y =CF 3 or Cl; R z =CH 3 , C.H. 2 CH 3 or Cl; or R y and R z Together: [ka] form; R a’ is H, F, Cl, CN, OH or OSO 3 - and; and R 1 and R 2 are each independently selected from the group consisting of hydrogen and methyl. 4. A method of treating AR+ / ER+ breast cancer in a subject comprising administering a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof. Method 2: A compound of formula I is reacted with compound II: [ka] The method of method 1, wherein Method 3. The compound of formula I is RAD140 (compound III): [ka] The method of method 1, wherein Method 4. The method of any of Methods 1-3, wherein administration is by oral route. Method 5. The method of any one of Methods 1 to 4, wherein the subject is female. Method 6. The method of Method 5, wherein said woman is a premenopausal woman. Method 7. The method of Method 5, wherein said woman is a postmenopausal woman. Method 8. The method of any of Methods 1-7, wherein said subject is treated in an adjuvant setting. Method 9. The method of any of Methods 1-7, wherein said subject has had disease progression following treatment with one or more endocrinological agents. Method 10. The method of Method 9, wherein said one or more endocrinological agents are selected from the group consisting of SERMs, SERDs, progestins, aromatase inhibitors, and combinations thereof. Method 11. The method of any of Methods 1-7, wherein the subject has had disease progression following treatment with one or more agents selected from the group consisting of a CDK4 / 6 inhibitor, an mTOR inhibitor, a BCL-2 inhibitor, a PI3K inhibitor, and combinations thereof. Method 12. The method according to any one of Methods 1 to 11, wherein the breast cancer is localized, advanced or metastatic breast cancer. Method 13. The method according to any one of Methods 3 to 12, wherein the RAD140 is administered at 10 to 500 mg, 10 to 250 mg, or 25 to 250 mg per day. Method 14. The method of Method 13, wherein said administration is once a day. Method 15. The method of any one of Methods 1 to 14, wherein the subject expresses ESR1 containing one or more mutations. Method 16. The method of method 15, wherein the mutation affects the binding affinity of a ligand compared to non-mutated ESR1. Method 17. The method of method 16, wherein the mutation results in a decrease in estradiol affinity for the mutated ESR1 compared to non-mutated ESR1. Method 18. The method of any of Methods 15 to 17, wherein the mutation signals through the ESR1 pathway in a ligand-dependent or ligand-independent manner. Method 19. The method of any of Methods 15 to 18, wherein the mutation results in a fusion protein comprising at least 10 consecutive amino acids from the sequence of non-mutated ESR1 and at least 10 consecutive amino acids from another human protein. Method 20. The method according to any one of Methods 15 to 19, wherein the mutation results in ESR1 lacking 10 or more consecutive amino acids from the amino acid sequence of the normal (non-mutated) ligand-binding domain of ESR1. Method 21. The method according to any one of Methods 15 to 20, wherein the mutation comprises one or more mutations selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X, and combinations thereof. Method 22. The method of any of Methods 1 to 21, wherein the treatment further comprises administration of a CDK4 / 6 inhibitor. Method 23. The CDK4 / 6 inhibitor has an IC of <100 nM for CDK4 and CDK6. 50 23. The method of method 22, comprising the steps of: Method 24. The method of any one of Methods 1 to 23, wherein the CDK4 / 6 inhibitor is selected from the group consisting of palbociclib, ribociclib, trilaciclib, and abemaciclib. Method 25. The method of any one of Methods 1 to 21, wherein the treatment further comprises administration of an mTOR inhibitor. Method 26. The method of Method 25, wherein said mTOR inhibitor is selected from the group consisting of sirolimus, temsirolimus, everolimus, ridafarolimus and MLN0128. Method 27. The method of any one of Methods 1 to 21, further comprising administration of a PI3K inhibitor. Method 28. The method of Method 27, wherein the PI3K inhibitor is BEZ235, GDC-0980, BKM120, GDC-0941, BYL719, GDC-0032, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223. Method 29. The method of any of Methods 1-21, further comprising administration of a PARP inhibitor. Method 30. The method of Method 29, wherein said PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186. Method 31. The method of any of Methods 1-21, further comprising administration of an MCL-1 inhibitor. Method 32. The method of Method 31, wherein said MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, sabutoclax or TW-37. Method 33. The method of any of Methods 1-21, further comprising administration of a BCL-2 inhibitor. Method 34. The method of Method 33, wherein said BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139 or S55746. Method 35. The method of any one of Methods 1-7 or 12-34, wherein said treatment is a first line treatment in a non-adjuvant setting. Method 36. A kit comprising an AR agonist according to any one of methods 1 to 3 and one or more agents selected from the group consisting of a PARP inhibitor, an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof. Method 37. A method of treating AR+ / ER+ breast cancer in a subject comprising administration of a steroidal or non-steroidal AR agonist together with one or more agents selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof. Method 38. The method of Method 37, wherein said AR agonist is a steroidal AR agonist. Method 39. The method of Method 38, wherein said AR agonist is a selective androgen receptor modulator. Method 40. The method of Method 39, wherein said selective androgen receptor modulator is selected from the group consisting of 2-chloro-4-[[(1R,2R)-2-hydroxy-2-methyl-cyclopentyl]amino]-3-methyl-benzonitrile, PF-06260414, enobosarm, BMS-564929, LGD-4033, AC-262356, JNJ-28330835, S-40503, GSK-2881078, AZD-3514, MK4541, LG121071, GLPG0492, NEP28, YK11, MK0773, ACP-105, LY-2452473, S-101479, S-40542, S-42 and LGD-3303. Method 41. The method of any of Methods 37-40, wherein said treatment is in an adjuvant setting. Method 42. The method of any of Methods 37-40, wherein said treatment is first line in a non-adjuvant setting. Method 43. The method of any of Methods 37-40, wherein the subject has had disease progression following treatment with a previous endocrinological therapy. Method 44. The method of any one of Methods 37-40 or 43, wherein the subject has had disease progression following treatment with an agent selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof. Method 45. The method of any one of Methods 37 to 44, wherein the subject is female. Method 46. The method of Method 45, wherein said woman is a premenopausal woman. Method 47. The method of Method 45, wherein said woman is a postmenopausal woman. Method 48. The method of any one of Methods 37 to 47, wherein the breast cancer is localized. Method 49. The method of any one of Methods 37 to 47, wherein the breast cancer is advanced or metastatic. Method 50. The method of any one of Methods 37 to 49, wherein the m-TOR inhibitor is sirolimus, temsirolimus, everolimus, ridafarolimus, or MLN0128. Method 51. The method of any of Methods 37 to 50, wherein the CDK4 / 6 inhibitor is palbociclib, ribociclib, trilaciclib, or abemaciclib. Method 52. The method of any one of Methods 37 to 51, wherein the PI3K inhibitor is BEZ235, GDC-0980, BKM120, GDC-0941, BYL719, GDC-0032, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223. Method 53. The method of any of Methods 37 to 52, wherein the PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186. Method 54. The method of any one of Methods 37 to 53, wherein the MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, sabutoclax, or TW-37. Method 55. Any of Methods 37 to 54, wherein the BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139 or S55746. Method 56. The method of any of Methods 37-55, wherein the active agents are administered concomitantly. Method 57. The method of any of Methods 35-56, wherein the active agents are administered in a coformulation. Method 58. A kit useful for treating breast cancer comprising an AR agonist or selective androgen receptor modulator and one or more agents selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor and combinations thereof. Method 59. A method of treating AR+ / ER+ breast cancer in a subject, wherein the subject has one or more ESR1 mutations, the method comprising administration of an AR agonist. Method 60. The method of method 59, wherein said AR agonist is non-steroidal. Method 61. The method of Method 60, wherein said AR agonist is a selective androgen receptor modulator. Method 62. The method of Method 61, wherein said selective androgen receptor modulator is selected from the group consisting of 2-chloro-4-[[(1R,2R)-2-hydroxy-2-methyl-cyclopentyl]amino]-3-methyl-benzonitrile, PF-06260414, enobosarm, BMS-564929, LGD-4033, AC-262356, JNJ-28330835, S-40503, GSK-2881078, AZD-3514, MK4541, LG121071, GLPG0492, NEP28, YK11, MK0773, ACP-105, LY-2452473, S-101479, S-40542, S-42 and LGD-3303. Method 63. The method of any of Methods 58 to 62, wherein the mutation affects the binding affinity of a ligand compared to non-mutated ESR1. Method 64. The method of any of Methods 59 to 63, wherein the mutation results in a decrease in estradiol affinity for the mutated ESR1 compared to non-mutated ESR1. Method 65. The method of any of Methods 59-64, wherein the mutation signals through the ESR1 pathway in a ligand-dependent or ligand-independent manner. Method 66. The method of any of Methods 59 to 65, wherein the mutation results in a fusion protein comprising at least 10 consecutive amino acids from the sequence of non-mutated ESR1 and at least 10 consecutive amino acids from another human protein. Method 67. The method according to any one of Methods 59 to 66, wherein the mutation results in ESR1 lacking 10 or more consecutive amino acids from the amino acid sequence of the normal (non-mutated) ligand-binding domain of ESR1. Method 68. The method of any of Methods 59 to 67, wherein the mutation is a fusion selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X, and combinations thereof. Method 69. The method of any of Methods 59-68, wherein said administration is by oral route. Method 70. The method of any of methods 59-69, wherein said treatment is in an adjuvant setting. Method 71. The method of any of Methods 59-69, wherein said treatment is first line in a non-adjuvant setting. Method 72. The method of any of Methods 59-69, wherein the subject has had disease progression following treatment with a previous endocrinological therapy. Method 73. The method of any one of Methods 59-69 or 72, wherein the subject has had disease progression following treatment with one or more agents selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof. Method 74. The method of any of Methods 59 to 73, wherein the subject is female. Method 75. The method of Method 74, wherein said woman is a premenopausal woman. Method 76. The method of Method 74, wherein said woman is a postmenopausal woman. Method 77. The method of any of Methods 59 to 76, wherein the breast cancer is localized. Method 78. The method of any of Methods 59 to 76, wherein the breast cancer is advanced or metastatic. Method 79. The method of any of Methods 59-78, wherein the treatment further comprises administration of a CDK4 / 6 inhibitor. Method 80. The CDK4 / 6 inhibitor has an IC of <100 nM against CDK4 and CDK6. 5080. The method of method 79, comprising: Method 81. The method of Method 79, wherein said CDK4 / 6 inhibitor is selected from the group consisting of palbociclib, ribociclib, trilaciclib and abemaciclib. Method 82. The method of any of methods 59-78, wherein the treatment further comprises administration of an mTOR inhibitor. Method 83. The method of Method 82, wherein said mTOR inhibitor is selected from the group consisting of sirolimus, temsirolimus, everolimus, ridafarolimus, and MLN0128. Method 84. The method of any of methods 59-78, further comprising administration of a PI3K inhibitor. Method 85. The method of Method 84, wherein the PI3K inhibitor is BEZ235, GDC-0980, BKM120, GDC-0941, BYL719, GDC-0032, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223. Method 86. The method of any of methods 59-78, further comprising administration of a PARP inhibitor. Method 87. The method of Method 86, wherein said PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186. Method 88. The method of any of methods 59-78, further comprising administration of an MCL-1 inhibitor. Method 89. The method of Method 88, wherein said MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, sabutoclax, or TW-37. Method 90. The method of any of methods 59-78, further comprising administration of a BCL-2 inhibitor. Method 91. The method of Method 90, wherein said BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139 or S55746. Method 92. The method of any of Methods 59-91, wherein said treatment is a first line treatment in a non-adjuvant setting. Method 93. 1) testing a subject for one or more ESR1 mutations; and 2) if the subject tests positive for one or more ESR1 mutations, treating the subject with any one of methods 1 to 35, 37 to 57, or 59 to 92. 4. A method of treating a subject for breast cancer comprising: Method 94. The method of Method 93, wherein the mutation results in a decrease in estradiol affinity for the mutated ESR1 compared to non-mutated ESR1. Method 95. The method of Method 93 or 94, wherein said mutation signals through the ESR1 pathway in a ligand-dependent or ligand-independent manner. Method 96. The method of any of Methods 93 to 95, wherein the mutation results in a fusion protein comprising at least 10 contiguous amino acids from the sequence of non-mutated ESR1 and at least 10 contiguous amino acids from another human protein. Method 97. The method according to any one of Methods 93 to 95, wherein the mutation results in ESR1 lacking 10 or more consecutive amino acids from the amino acid sequence of the normal (non-mutated) ligand-binding domain of ESR1. Method 98. The method of Method 93, wherein the mutation is a fusion selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X, and combinations thereof. Method 99. A method described in any one of Methods 1-35, 37-57, 59-98, wherein the subject is first tested for a baseline level of ZBTB16 mRNA or protein expression, and then tested again after a period of treatment for the level of ZBTB16 mRNA or protein expression, and if the level increases above baseline, it is recommended that the subject continue treatment. Method 100. The method of Method 99, wherein said treatment period is daily administration of the AR agonist for at least 3 days. Method 101. The method of Method 100, wherein said period of time is daily administration of the AR agonist for at least one week. Method 102. The method of any of Methods 99-101, wherein the ratio of post-treatment level to pre-treatment level is >3. Method 103. The method of method 102, wherein said ratio is >10. Method 104. The method of method 103, wherein said ratio is >50.

[0093] The aspects of the present invention include the following. [1] A subject is administered a dose of Formula I [ka] (In the formula, R x =CN; R y =CF 3 or Cl; R z =CH 3 , C.H. 2 CH 3 or Cl; or R y and R z Together: [ka] form; R a’ is H, F, Cl, CN, OH or OSO 3 - and; and R1 and R 2 are each independently selected from the group consisting of hydrogen and methyl. 4. A method of treating AR+ / ER+ breast cancer in a subject comprising administering a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof. [2] The compound of formula I is a compound II: [ka] The method according to [1], [3] The compound of formula I is RAD140 (compound III): [ka] The method according to [1], [4] The method according to any one of [1] to [3], wherein administration is by oral route. [5] The method described in any one of [1] to [4], wherein the subject is a female. [6] The method described in [5], wherein the woman is a premenopausal woman. [7] The method described in [5], wherein the woman is a postmenopausal woman. [8] The method of any of [1] to [7], wherein the subject is treated in an adjuvant setting. [9] The method of any of [1] to [7], wherein the subject has had disease progression following treatment with one or more endocrinological agents.

[10] The method of [9], wherein the one or more endocrinological agents are selected from the group consisting of SERMs, SERDs, progestins, aromatase inhibitors, and combinations thereof.

[11] The method of any of [1] to [7], wherein the subject has had disease progression after treatment with one or more agents selected from the group consisting of CDK4 / 6 inhibitors, mTOR inhibitors, BCL-2 inhibitors, PI3K inhibitors, and combinations thereof.

[12] The method described in any of [1] to

[11] , wherein the breast cancer is localized, progressive or metastatic breast cancer.

[13] The method described in any of [3] to

[12] , wherein the RAD140 is administered at 10 to 500 mg, 10 mg to 250 mg, or 25 mg to 250 mg per day.

[14] The method of

[13] , wherein the administration is once a day.

[15] The method described in any one of [1] to

[14] , wherein the subject expresses ESR1 containing one or more mutations.

[16] The method of

[15] , wherein the mutation affects the binding affinity of a ligand compared to non-mutated ESR1.

[17] The method of

[16] , wherein the mutation results in reduced estradiol affinity for the mutated ESR1 compared to non-mutated ESR1.

[18] The method according to any one of

[15] to

[17] , wherein the mutation transmits signals via the ESR1 pathway in a ligand-dependent or ligand-independent manner.

[19] A method described in any of

[15] to

[18] , wherein the mutation results in a fusion protein containing at least 10 consecutive amino acids from the sequence of non-mutated ESR1 and at least 10 consecutive amino acids from another human protein.

[20] The method described in any of

[15] to

[19] , wherein the mutation results in ESR1 lacking 10 or more consecutive amino acids from the amino acid sequence of the normal (non-mutated) ligand-binding domain of ESR1.

[21] The method described in any one of

[15] to

[20] , wherein the mutation includes one or more mutations selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X, and combinations thereof.

[22] The method described in any of [1] to

[21] , wherein the treatment further comprises administration of a CDK4 / 6 inhibitor.

[23] The CDK4 / 6 inhibitor has an IC of <100 nM for CDK4 and CDK6. 50 The method according to

[22] , comprising:

[24] The method of any of [1] to

[23] , wherein the CDK4 / 6 inhibitor is selected from the group consisting of palbociclib, ribociclib, trilaciclib and abemaciclib.

[25] The method described in any of [1] to

[21] , wherein the treatment further comprises administration of an mTOR inhibitor.

[26] The method of

[25] , wherein the mTOR inhibitor is selected from the group consisting of sirolimus, temsirolimus, everolimus, ridafarolimus and MLN0128.

[27] The method described in any of [1] to

[21] , further comprising administration of a PI3K inhibitor.

[28] The method of

[27] , wherein the PI3K inhibitor is BEZ235, GDC-0980, BKM120, GDC-0941, BYL719, GDC-0032, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223.

[29] The method described in any of [1] to

[21] , further comprising administration of a PARP inhibitor.

[30] The method of

[29] , wherein the PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186.

[31] The method of any of [1] to

[21] , further comprising administration of an MCL-1 inhibitor.

[32] The method of

[31] , wherein the MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, sabutoclax or TW-37.

[33] The method described in any of [1] to

[21] , further comprising administration of a BCL-2 inhibitor.

[34] The method of

[33] , wherein the BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139 or S55746.

[35] The method of any one of [1] to [7] or

[12] to

[34] , wherein the treatment is a first-line treatment in a non-adjuvant setting.

[36] A kit comprising an AR agonist according to any one of [1] to [3] and one or more agents selected from the group consisting of PARP inhibitors, mTOR inhibitors, CDK4 / 6 inhibitors, PI3K inhibitors, BCL2 inhibitors, MCL-1 inhibitors, and combinations thereof.

[37] A method for treating AR+ / ER+ breast cancer in a subject, comprising administering a steroidal or non-steroidal AR agonist together with one or more agents selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof.

[38] The method of

[37] , wherein the AR agonist is a steroidal AR agonist.

[39] The method of

[38] , wherein the AR agonist is a selective androgen receptor modulator.

[40] The method of

[39] , wherein the selective androgen receptor modulator is selected from the group consisting of 2-chloro-4-[[(1R,2R)-2-hydroxy-2-methyl-cyclopentyl]amino]-3-methyl-benzonitrile, PF-06260414, enobosarm, BMS-564929, LGD-4033, AC-262356, JNJ-28330835, S-40503, GSK-2881078, AZD-3514, MK4541, LG121071, GLPG0492, NEP28, YK11, MK0773, ACP-105, LY-2452473, S-101479, S-40542, S-42 and LGD-3303.

[41] The method of any of

[37] to

[40] , wherein the treatment is in an adjuvant setting.

[42] The method of any of

[37] to

[40] , wherein the treatment is first-line in a non-adjuvant setting.

[43] The method of any of

[37] to

[40] , wherein the subject has had disease progression after treatment with a previous endocrinological therapy.

[44] The method of any one of

[37] to

[40] or

[43] , wherein the subject has had disease progression after treatment with one or more agents selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof.

[45] The method described in any one of

[37] to

[44] , wherein the subject is a female.

[46] The method of

[45] , wherein the woman is a premenopausal woman.

[47] The method of

[45] , wherein the woman is a postmenopausal woman.

[48] ​​The method described in any one of

[37] to

[47] , wherein the breast cancer is localized.

[49] The method described in any one of

[37] to

[47] , wherein the breast cancer is progressive or metastatic.

[50] The method described in any of

[37] to

[49] , wherein the m-TOR inhibitor is sirolimus, temsirolimus, everolimus, ridafarolimus or MLN0128.

[51] The method described in any of

[37] to

[50] , wherein the CDK4 / 6 inhibitor is palbociclib, ribociclib, trilaciclib or abemaciclib.

[52] The method of any of

[37] to

[51] , wherein the PI3K inhibitor is BEZ235, GDC-0980, BKM120, GDC-0941, BYL719, GDC-0032, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223.

[53] The method of any of

[37] to

[52] , wherein the PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186.

[54] The method according to any one of

[37] to

[53] , wherein the MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, sabutoclax or TW-37.

[55] The method described in any of

[37] to

[54] , wherein the BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139 or S55746.

[56] The method of any one of

[37] to

[55] , wherein an active agent is administered together.

[57] The method of any one of

[35] to

[56] , wherein the active agent is administered in a coformulation.

[58] A kit useful for treating breast cancer, comprising an AR agonist or selective androgen receptor modulator and one or more agents selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof.

[59] A method of treating AR+ / ER+ breast cancer in a subject, the subject having one or more ESR1 mutations, the method comprising administration of an AR agonist.

[60] The method of

[59] , wherein the AR agonist is non-steroidal.

[61] The method of

[60] , wherein the AR agonist is a selective androgen receptor modulator.

[62] The method of

[61] , wherein the selective androgen receptor modulator is selected from the group consisting of 2-chloro-4-[[(1R,2R)-2-hydroxy-2-methyl-cyclopentyl]amino]-3-methyl-benzonitrile, PF-06260414, enobosarm, BMS-564929, LGD-4033, AC-262356, JNJ-28330835, S-40503, GSK-2881078, AZD-3514, MK4541, LG121071, GLPG0492, NEP28, YK11, MK0773, ACP-105, LY-2452473, S-101479, S-40542, S-42 and LGD-3303.

[63] The method described in any one of

[59] to

[62] , wherein the mutation affects the binding affinity of a ligand compared to non-mutated ESR1.

[64] A method described in any of

[59] to

[63] , wherein the mutation results in reduced estradiol affinity for the mutated ESR1 compared to non-mutated ESR1.

[65] The method of any one of

[59] to

[64] , wherein the mutation signals via the ESR1 pathway in a ligand-dependent or ligand-independent manner.

[66] A method described in any of

[59] to

[65] , wherein the mutation results in a fusion protein containing at least 10 consecutive amino acids from the sequence of non-mutated ESR1 and at least 10 consecutive amino acids from another human protein.

[67] A method described in any of

[59] to

[66] , wherein the mutation results in ESR1 lacking 10 or more consecutive amino acids from the amino acid sequence of the normal (non-mutated) ligand-binding domain of ESR1.

[68] The method described in any of

[59] to

[67] , wherein the mutation is a fusion selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X and combinations thereof.

[69] The method of any one of

[59] to

[68] , wherein the administration is by oral route.

[70] The method of any of

[59] to

[69] , wherein the treatment is in an adjuvant setting.

[71] The method of any of

[59] to

[69] , wherein the treatment is first-line in a non-adjuvant setting.

[72] The method of any of

[59] to

[69] , wherein the subject has had disease progression following treatment with a previous endocrinological therapy.

[73] The method of any one of

[59] to

[69] or

[72] , wherein the subject has had disease progression following treatment with one or more agents selected from the group consisting of an mTOR inhibitor, a CDK4 / 6 inhibitor, a PI3K inhibitor, a PARP inhibitor, a BCL2 inhibitor, an MCL-1 inhibitor, and combinations thereof.

[74] The method described in any one of

[59] to

[73] , wherein the subject is female.

[75] The method of

[74] , wherein the woman is a premenopausal woman.

[76] The method of

[74] , wherein the woman is a postmenopausal woman.

[77] The method described in any one of

[59] to

[76] , wherein the breast cancer is localized.

[78] The method described in any one of

[59] to

[76] , wherein the breast cancer is advanced or metastatic.

[79] A method described in any of

[59] to

[78] , wherein the treatment further comprises administration of a CDK4 / 6 inhibitor.

[80] The CDK4 / 6 inhibitor has an IC of <100 nM for CDK4 and CDK6. 50 The method of

[79] , comprising:

[81] The method of

[79] , wherein the CDK4 / 6 inhibitor is selected from the group consisting of palbociclib, ribociclib, trilaciclib and abemaciclib.

[82] The method described in any of

[59] to

[78] , wherein the treatment further comprises administration of an mTOR inhibitor.

[83] The method of

[82] , wherein the mTOR inhibitor is selected from the group consisting of sirolimus, temsirolimus, everolimus, ridafarolimus and MLN0128.

[84] The method described in any of

[59] to

[78] , further comprising administration of a PI3K inhibitor.

[85] The method of

[84] , wherein the PI3K inhibitor is BEZ235, GDC-0980, BKM120, GDC-0941, BYL719, GDC-0032, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223.

[86] The method described in any of

[59] to

[78] , further comprising administration of a PARP inhibitor.

[87] The method of

[86] , wherein the PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186.

[88] The method described in any of

[59] to

[78] , further comprising administration of an MCL-1 inhibitor.

[89] The method of

[88] , wherein the MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, sabutoclax or TW-37.

[90] The method described in any of

[59] to

[78] , further comprising administration of a BCL-2 inhibitor.

[91] The method of

[90] , wherein the BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139 or S55746.

[92] The method of any of

[59] to

[91] , wherein the treatment is a first-line treatment in a non-adjuvant setting.

[93] 1) testing a subject for one or more ESR1 mutations; and 2) treating the subject with the method according to any one of [1] to

[35] ,

[37] to

[57] , or

[59] to

[92] if the subject tests positive for one or more ESR1 mutations. 4. A method of treating a subject for breast cancer comprising:

[94] The method of

[93] , wherein the mutation results in reduced estradiol affinity for the mutated ESR1 compared to non-mutated ESR1.

[95] The method of

[93] or

[94] , wherein the mutation signals through the ESR1 pathway in a ligand-dependent or ligand-independent manner.

[96] A method described in any of

[93] to

[96] , wherein the mutation results in a fusion protein containing at least 10 consecutive amino acids from the sequence of non-mutated ESR1 and at least 10 consecutive amino acids from another human protein.

[97] A method described in any of

[93] to

[96] , wherein the mutation results in ESR1 lacking 10 or more consecutive amino acids from the amino acid sequence of the normal (non-mutated) ligand-binding domain of ESR1.

[98] The method of

[93] , wherein the mutation is a fusion selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X and combinations thereof.

[99] The method of any one of [1]-

[35] ,

[37] -

[57] ,

[59] -

[98] , wherein the subject is first tested for a baseline level of ZBTB16 mRNA or protein expression, and then tested again after a period of treatment for the level of ZBTB16 mRNA or protein expression, and if the level increases above baseline, it is recommended that the subject continue treatment.

[0100]

[99] The method of claim 1, wherein the treatment period is daily administration of the AR agonist for at least 3 days.

[0101] The method of

[0100] , wherein the period is daily administration of an AR agonist for at least one week.

[0102] A method described in any of

[99] to

[0101] , wherein the ratio of post-treatment level to pre-treatment level is >3.

[0103] The method described in

[0102] , wherein the ratio is >10.

[0104] The method described in

[0103] , wherein the ratio is >50.

Claims

1. 1. A medicament for treating AR+ / ER+ / HER2- breast cancer in a subject, comprising compound III: 【Chemistry 1】 or a pharma- ceutically acceptable salt or a pharma- ceutically acceptable solvate thereof.

2. The method of claim 1, wherein the compound is administered by oral route.

3. (i) the subject is treated in an adjuvant setting; (ii) the subject has disease progression following treatment with one or more endocrinological agents; and / or (iii) The pharmaceutical agent of claim 1 or 2, wherein the subject has disease progression after treatment with one or more agents selected from the group consisting of CDK4 / 6 inhibitors, mTOR inhibitors, BCL-2 inhibitors, PI3K inhibitors, and combinations thereof.

4. The pharmaceutical composition of claim 3, wherein the one or more endocrinological agents are selected from the group consisting of SERMs, SERDs, progestins, aromatase inhibitors, and combinations thereof.

5. The method of any one of claims 1 to 4, wherein the breast cancer is resistant to at least one previous treatment.

6. The method of any one of claims 1 to 5, wherein the subject has progressed on previous endocrinological therapy.

7. The method of claim 1 , wherein the subject is a candidate for first-line treatment.

8. The method of any one of claims 1 to 7, wherein the compound is administered at 10 to 500 mg, 10 mg to 250 mg, or 25 mg to 250 mg per day.

9. The pharmaceutical composition of claim 8, wherein the administration is once a day.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the subject expresses ESR1 containing one or more mutations.

11. The pharmaceutical composition of claim 10, wherein the mutation comprises one or more mutations selected from the group consisting of ESR1-AKAP12, ESR1-CCDC170, ESR1-YAP1, ESR1-POLH, ESR1-PCDH11X, and combinations thereof.

12. The pharmaceutical composition of any one of claims 1 to 11, wherein the subject expresses a PI3K mutation.

13. The pharmaceutical composition according to claim 12, wherein the PI3K mutation is PIK3CA.

14. The pharmaceutical composition of any one of claims 1 to 13, wherein the treatment further comprises administration of a CDK4 / 6 inhibitor.

15. The pharmaceutical composition of claim 14, wherein the CDK4 / 6 inhibitor is selected from the group consisting of palbociclib, ribociclib, trilaciclib, abemaciclib and AMG925.

16. The pharmaceutical composition of any one of claims 1 to 14, wherein the treatment further comprises administration of a second therapeutic agent that targets the PI3K / AKT / mTOR pathway.

17. The pharmaceutical described in claim 16, wherein the second therapeutic agent is an mTOR inhibitor and / or a PI3K inhibitor.

18. The pharmaceutical composition of claim 17, wherein the mTOR inhibitor is selected from the group consisting of sirolimus, temsirolimus, everolimus, ridafarolimus and MLN0128, and / or the PI3K inhibitor is BEZ235, GDC-0980, GDC-0941, BYL719, MK2206, GDC-0068, GSK2110183, GSK2141795, AZD5363, AZD2014, MLN0128 or CC-223.

19. The treatment, (i) a PARP inhibitor, (ii) an MCL-1 inhibitor, and / or (iii) a BCL-2 inhibitor, The method of any one of claims 1 to 16, further comprising administering:

20. The following: (i) the PARP inhibitor is talazoparib, veliparib, niraparib, beigene290, E7449, KX01, ABT767, CK102, JPI289, KX02, IMP4297, SC10914, NT125, PJ34, VPI289 or ANG-3186; (ii) the MCL-1 inhibitor is 7-(5-((4-(4-(N,N-dimethylsulfamoyl)piperazin-1-yl)phenoxy)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1-(2-morpholinoethyl)-3-(3-(naphthalen-1-yloxy)propyl)-1H-indole-2-carboxylic acid, S63845, omasataxin, seliciclib, UMI-77, AT101, or sabutoclax; and / or (iii) the BCL-2 inhibitor is venetoclax, navitoclax, ABT737, G3139 or S55746; The pharmaceutical composition of claim 19, wherein at least one of the above is satisfied.

21. The pharmaceutical composition of any one of claims 1 to 15, wherein the treatment further comprises administration of a therapeutic agent selected from the group consisting of rapamycin, everolimus, ridaforolimus, temsirolimus, MLN0128, CC115, CC223, OSI-027, AZD8055, AZD2014, GDC-0980, SAR245409, LY3023414, NVP-BEZ235, NVP-BGT226, SF1126 and PKI-587.

22. The pharmaceutical composition of any one of claims 1 to 13, wherein the treatment further comprises administration of a therapeutic agent selected from the group consisting of everolimus, BYL719, GDC-0068, palbociclib, AZD5363 and abemaciclib, and combinations thereof.

23. The pharmaceutical composition of any one of claims 1 to 15, wherein the treatment further comprises administration of everolimus.

24. The pharmaceutical composition of any one of claims 1 to 15, wherein the treatment further comprises administration of BYL719.

25. The pharmaceutical composition according to any one of claims 1 to 24, wherein the subject is a female.

26. The pharmaceutical composition described in claim 25, wherein the female is a premenopausal female or a postmenopausal female.

27. The method according to any one of claims 1 to 25, wherein the breast cancer is localized, advanced or metastatic.

Images