Lasofoxifene treatment for ER+ breast cancer

Abstract

To provide lasofoxifene treatment of ER+ breast cancer.SOLUTION: The disclosure provides methods for treating estrogen receptor positive (ER+) cancer in women with an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. The disclosure also includes detection of Estrogen Receptor 1 (ESR1) gene mutations that lead to endocrine resistance and treatment of endocrine resistant ER+ cancers. In one aspect, the methods further comprise selecting, for treatment, a patient who has been diagnosed with estrogen receptor positive (ER+) breast cancer.SELECTED DRAWING: None

Description

[Technical Field]

[0001] 1. CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application Nos. 62 / 502,299, filed May 5, 2017; 62 / 457,759, filed February 10, 2017; and 62 / 406,859, filed October 11, 2016, each of which is incorporated herein by reference in its entirety.

[0002] 2. Sequence Listing This application contains a Sequence Listing, which was submitted via EFS-Web and is incorporated herein by reference in its entirety. This ASCII copy was created on October 9, 2017, is named 33498PCT_CRF_sequencelisting.txt, and is 2,119 bytes in size. [Background technology]

[0003] Estrogen receptor positive (ER) + ) breast cancer is a group of breast cancers that express estrogen receptor alpha (ERα). Approximately 70% of breast cancers are ERα-expressing. + and is therefore treated by endocrine therapy, which reduces estrogen levels or blocks estrogen signaling to inhibit ER. + It has led to significant improvements in outcomes for women with breast cancer, however its effectiveness is limited by intrinsic and acquired endocrine resistance.

[0004] Recent research has shown that ER +We present evidence for the temporal selection of functional estrogen receptor 1 (ESR1) gene mutations as a potential driver of endocrine resistance during breast cancer progression. See Jeselsohn et al., Clinical Cancer Research 20(7):1757-1767 (2014). Mutations in ESR1, the gene encoding ERα, alter the conformation of the ERα protein, increasing its interaction with its coactivators, promoting the active form of the receptor in the absence of hormone and helping tumor cells evade hormone treatment. See Thomas and Gustafsson, Trends in Endocrinology and Metabolism 26(9):467-476 (2015).

[0005] Therefore, there remains a need to develop new therapeutic strategies that are effective in treating tumors with mutations in ESR1 and that can therefore be used to treat breast cancer patients who have developed or are at risk of developing endocrine resistance. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Jeselsohn et al., Clinical Cancer Research (2014) 20(7):1757-1767 [Non-patent document 2] Thomas and Gustafsson, Trends in Endocrinology and Metabolism (2015) 26(9):467-476 Summary of the Invention [Means for solving the problem]

[0007] We created ERα expression constructs expressing four ESR1 mutations, Y537S, Y537N, Y537C, and D538G, in the ligand-binding domain (LBD) of the ERα protein and introduced these expression constructs into cells in culture. These mutations were found to be involved in the regulation of ERα expression in endocrine-treated ERα-treated mice. + Found in patients with metastatic breast cancer. Jeselsohn et al., Nature Reviews Clinical Oncology 12(10):573-583 (2015); Jeselsohn et al., Clinical Cancer Research 20(7):1757-1767 (2014); Robinson et al., Nature Genetics 45(12):1446-1451 (2013); Thomas and Gustafsson, Trends in See Endocrinology and Metabolism 26(9):467-476 (2015); and Toy et al., Nature Genetics 45(12):1439-1445 (2013).

[0008] Using estrogen receptor-responsive reporter constructs, we confirmed that all mutants were constitutively active compared with wild-type ERα in ovarian and breast cancer cell lines. We then treated the cells with lasofoxifene, a selective ER modulator (SERM), and found that lasofoxifene effectively inhibited the transcriptional activity of the ERα LBD mutants in a dose-responsive manner at clinically achievable concentrations.

[0009] In a second series of experiments, we confirmed that lasofoxifene, at clinically achievable concentrations, was able to reduce the viability of the breast cancer cell line MCF7 stably transfected with either the Y537S or D538G ESR1 mutant receptor.

[0010] Thus, in a first aspect, a method of treating locally advanced or metastatic breast cancer in a woman is provided. The method comprises administering estrogen receptor-positive (ER) +2.) selecting for treatment a patient diagnosed with locally advanced or metastatic breast cancer, and administering to the selected patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0011] In various embodiments, the selected patient has been previously treated with one or more lines of endocrine therapy, hi certain embodiments, the patient has been previously treated with multiple lines of endocrine therapy.

[0012] In some embodiments, the endocrine therapy the patient has previously been treated with is a selective ER modulator (SERM). In certain embodiments, the SERM is tamoxifen, raloxifene, bazedoxifene, toremifene, or ospemifene.

[0013] In some embodiments, the endocrine therapy the patient has previously been treated with is a selective ER degrading drug (SERD). In certain embodiments, the SERD is fulvestrant, RAD1901, ARN-810 (GDC-0810), or AZD9496.

[0014] In some embodiments, the endocrine therapy the patient was previously treated with is an aromatase inhibitor. In certain embodiments, the aromatase inhibitor is exemestane (Aromasin®), letrozole (Femara®), or anastrozole (Arimidex®).

[0015] In some embodiments, the patient has disease progression after endocrine therapy. In some embodiments, the patient is resistant to endocrine therapy.

[0016] In various embodiments, the patient's cancer has at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. In some embodiments, the patient has previously been determined to have at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. In certain embodiments, the method further comprises the preceding step of determining that the patient has at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene.

[0017] In some embodiments, the at least one gain-of-function missense mutation is at any one of amino acids D538, Y537, L536, P535, V534, S463, V392, or E380.

[0018] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid D538. In some preferred embodiments, the mutation is D538G.

[0019] In certain embodiments, at least one gain-of-function missense mutation is at amino acid Y537. In some embodiments, the mutation is Y537S, Y537N, Y537C, or Y537Q. In some preferred embodiments, the mutation is Y537C.

[0020] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid L536. In some embodiments, the mutation is L536R or L536Q.

[0021] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid P535. In some embodiments, the mutation is P535H.

[0022] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid V534. In some embodiments, the mutation is V534E.

[0023] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid S463. In some embodiments, the mutation is S463P.

[0024] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid V392. In some embodiments, the mutation is V392I.

[0025] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid E380. In some embodiments, the mutation is E380Q.

[0026] In some embodiments, the patient's serum estradiol level is at least 0.35 ng / dL. In some embodiments, the patient's serum estradiol level is from about 0.30 ng / dL to about 0.35 ng / dL. In some embodiments, the patient's serum estradiol level is from about 0.25 ng / dL to about 0.30 ng / dL.

[0027] In various embodiments, lasofoxifene is administered in selected ER +It is administered as lasofoxifene tartrate to patients with locally advanced or metastatic breast cancer. In various embodiments, lasofoxifene is administered orally, intravenously, transdermally, topically, or via vaginal ring administration. In certain embodiments, lasofoxifene is administered orally. In some of these embodiments, lasofoxifene is administered orally (os) (po) at about 0.5 mg / day to about 10 mg / day. In certain embodiments, lasofoxifene is administered orally at about 0.5 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day. In certain embodiments, lasofoxifene is administered orally at about 5 mg / day. In various embodiments, lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month.

[0028] In certain embodiments, the method further comprises treating the patient with at least one additional endocrine therapy. In some embodiments, the patient is treated with the additional endocrine therapy at the original dose. In some other embodiments, the patient is treated with the additional endocrine therapy at a dose higher than the original dose. In certain embodiments, the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. In certain embodiments, the additional endocrine therapy is treatment with a selective ER degrader (SERD). In certain embodiments, the additional endocrine therapy is treatment with an aromatase inhibitor.

[0029] In various embodiments, the method comprises: +The method further comprises administering to the patient with locally advanced or metastatic breast cancer an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. In certain embodiments, the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. In some embodiments, the method further comprises administering to the patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. In certain embodiments, the mTOR inhibitor is everolimus. In some embodiments, the method further comprises administering to the patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. In some embodiments, the method further comprises administering to the patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. In certain embodiments, the HER2 inhibitor is trastuzumab (Herceptin®) or ado-trastuzumab emtansine (Kadcyla®). In some embodiments, the method further comprises administering to the patient an effective amount of a histone deacetylase (HDAC) inhibitor. In some of these embodiments, the HDAC inhibitor is selected from the group consisting of vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), abexinostat (P CI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), kevetrin, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rocilinostat (ACY-1215), or sulforaphane.In some embodiments, the method further comprises administering to the patient an effective amount of a checkpoint inhibitor. In some of these embodiments, the checkpoint inhibitor is an antibody specific for programmed death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). In certain embodiments, the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). In certain embodiments, the CTLA-4 antibody is ipilimumab (Yervoy®). In some embodiments, the method further comprises administering to the patient an effective amount of a cancer vaccine.

[0030] In some embodiments, the patient is premenopausal. In certain embodiments, the patient has locally advanced or metastatic ER+ / HER2- breast cancer. In some of these embodiments, the patient is progressing during the first hormone treatment, such as a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI combined with a CDK4 / 6 inhibitor, or fulvestrant combined with a CDK4 / 6 inhibitor.

[0031] In some embodiments, the patient is perimenopausal. In certain embodiments, the patient has locally advanced or metastatic ER+ / HER2- breast cancer. In some of these embodiments, the patient is progressing during the first hormone treatment, such as a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI combined with a CDK4 / 6 inhibitor, or fulvestrant combined with a CDK4 / 6 inhibitor.

[0032] In some embodiments, the patient is postmenopausal. In certain embodiments, the patient has locally advanced or metastatic ER+ / HER2- breast cancer. In some of these embodiments, the patient has progressed during the first hormonal treatment with a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI combined with a CDK4 / 6 inhibitor, or fulvestrant combined with a CDK4 / 6 inhibitor.

[0033] In another aspect, a method of treating primary breast cancer in a woman is provided. The method comprises treating primary breast cancer in an estrogen receptor positive (ER) cell line. + ) selecting a patient diagnosed with primary breast cancer for treatment, and administering to the selected patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0034] In various embodiments, lasofoxifene is administered in selected ER + It is administered to patients with primary breast cancer as lasofoxifene tartrate. In some embodiments, lasofoxifene is administered orally, intravenously, transdermally, topically, or via vaginal ring administration. In certain embodiments, lasofoxifene is administered orally. In some of these embodiments, lasofoxifene is administered orally at about 0.5 mg / day to about 10 mg / day. In certain embodiments, lasofoxifene is administered orally at about 0.5 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day. In certain embodiments, lasofoxifene is administered orally at about 5 mg / day. In various embodiments, lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month.

[0035] In various embodiments, ER +The method for treating primary breast cancer further comprises treating the patient with at least one additional endocrine therapy. In some embodiments, the patient is treated with the additional endocrine therapy at the original dose. In some other embodiments, the patient is treated with the additional endocrine therapy at a dose higher than the original dose. In certain embodiments, the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. In certain embodiments, the additional endocrine therapy is treatment with a selective ER degrader (SERD). In certain embodiments, the additional endocrine therapy is treatment with an aromatase inhibitor.

[0036] In various embodiments, the method comprises: +The method further comprises administering to the patient with primary breast cancer an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. In certain embodiments, the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. In some embodiments, the method further comprises administering to the patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. In certain embodiments, the mTOR inhibitor is everolimus. In some embodiments, the method further comprises administering to the patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. In some embodiments, the method further comprises administering to the patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. In certain embodiments, the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). In some embodiments, the method further comprises administering to the patient an effective amount of a histone deacetylase (HDAC) inhibitor. In some of these embodiments, the HDAC inhibitor is selected from the group consisting of vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), and the like. ), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), kevetlin, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. In some embodiments, the method further comprises administering to the patient an effective amount of a checkpoint inhibitor.In some of these embodiments, the checkpoint inhibitor is an antibody specific for programmed death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). In certain embodiments, the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). In certain embodiments, the CTLA-4 antibody is ipilimumab (Yervoy®). In some embodiments, the method further comprises administering to the patient an effective amount of a cancer vaccine.

[0037] In certain embodiments, the patient is premenopausal. In certain embodiments, the patient is peri-menopausal. In certain embodiments, the patient is post-menopausal.

[0038] In another aspect, a method for adjuvant therapy of estrogen receptor positive (ER+) breast cancer is provided, comprising administering to a patient who has received primary treatment for ER+ breast cancer an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof, in combination with an aromatase inhibitor.

[0039] In some embodiments, lasofoxifene is administered continuously during administration of the aromatase inhibitor. In some embodiments, lasofoxifene is administered cyclically during administration of the aromatase inhibitor. In certain embodiments, the dosing regimen of lasofoxifene is different from the dosing regimen of the aromatase inhibitor.

[0040] In various embodiments, lasofoxifene is administered as adjuvant therapy in combination with an aromatase inhibitor as lasofoxifene tartrate. In some embodiments, the aromatase inhibitor is exemestane (Aromasin®), letrozole (Femara®), or anastrozole (Arimidex®). In some embodiments, lasofoxifene is administered orally, intravenously, transdermally, topically, or via vaginal ring administration. In certain embodiments, lasofoxifene is administered orally. In some of these embodiments, lasofoxifene is administered at about 0.5 mg / day to about 10 mg / day orally. In certain embodiments, lasofoxifene is administered at about 0.5 mg / day to about 5 mg / day orally. In certain embodiments, lasofoxifene is administered at about 1 mg / day to about 5 mg / day orally. In certain embodiments, lasofoxifene is orally administered at about 1 mg / day.In certain embodiments, lasofoxifene is orally administered at about 5 mg / day.In various embodiments, lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month.

[0041] In various embodiments, the method of adjuvant therapy for estrogen receptor positive (ER+) breast cancer further comprises treating the patient with at least one additional endocrine therapy. In certain embodiments, the additional endocrine therapy is treatment with a selective ER degrading drug (SERD).

[0042] In various embodiments, the method of adjuvant therapy for estrogen receptor-positive (ER+) breast cancer further comprises administering to the patient an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. In certain embodiments, the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. In some embodiments, the method further comprises administering to the patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. In certain embodiments, the mTOR inhibitor is everolimus. In some embodiments, the method further comprises administering to the patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. In some embodiments, the method further comprises administering to the patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. In certain embodiments, the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). In some embodiments, the method further comprises administering to the patient an effective amount of a histone deacetylase (HDAC) inhibitor. In some of these embodiments, the HDAC inhibitor is selected from the group consisting of vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103 ), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), kevetlin, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. In some embodiments, the method further comprises administering to the patient an effective amount of a checkpoint inhibitor.In some of these embodiments, the checkpoint inhibitor is an antibody specific for programmed death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). In certain embodiments, the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). In certain embodiments, the CTLA-4 antibody is ipilimumab (Yervoy®). In some embodiments, the method further comprises administering to the patient an effective amount of a cancer vaccine.

[0043] In some embodiments, lasofoxifene is administered in an amount and on a schedule sufficient to improve bone mass. In some embodiments, lasofoxifene is administered in an amount and on a schedule sufficient to improve the symptoms of VVA.

[0044] In certain embodiments, the patient is premenopausal. In certain embodiments, the patient is peri-menopausal. In certain embodiments, the patient is post-menopausal.

[0045] In another aspect, a method is provided for treating cancer other than breast cancer in a woman. The method comprises treating estrogen receptor positive (ER) cancer other than breast cancer. + ) cancer and having at least one gain-of-function mutation in the estrogen receptor 1 (ESR1) gene, and selecting for treatment a patient with the cancer; and administering to the selected patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. In some embodiments, the patient has ER cancer. + In some other embodiments, the patient has been diagnosed with ovarian cancer. + He has been diagnosed with lung cancer.

[0046] In various embodiments, lasofoxifene is used to treat ER cancer other than breast cancer. +It is administered as lasofoxifene tartrate to selected patients with cancer. In some embodiments, lasofoxifene is administered orally, intravenously, transdermally, topically, or via vaginal ring administration. In certain embodiments, lasofoxifene is administered orally. In some of these embodiments, lasofoxifene is administered orally at about 0.5 mg / day to about 10 mg / day. In certain embodiments, lasofoxifene is administered orally at about 0.5 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day. In certain embodiments, lasofoxifene is administered orally at about 5 mg / day. In various embodiments, lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month.

[0047] In various embodiments, ER other than breast cancer. + The method for treating cancer further comprises treating the patient with at least one additional endocrine therapy. In some embodiments, the patient is treated with the additional endocrine therapy at the original dose. In some other embodiments, the patient is treated with the additional endocrine therapy at a dose higher than the original dose. In certain embodiments, the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. In certain embodiments, the additional endocrine therapy is treatment with a selective ER degrader (SERD). In certain embodiments, the additional endocrine therapy is treatment with an aromatase inhibitor.

[0048] In various embodiments, the method comprises administering to a patient a cancer other than breast cancer. +The method further comprises administering to the patient having cancer an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. In certain embodiments, the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. In some embodiments, the method further comprises administering to the patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. In certain embodiments, the mTOR inhibitor is everolimus. In some embodiments, the method further comprises administering to the patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. In some embodiments, the method further comprises administering to the patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. In certain embodiments, the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). In some embodiments, the method further comprises administering to the patient an effective amount of a histone deacetylase (HDAC) inhibitor. In some of these embodiments, the HDAC inhibitor is selected from the group consisting of vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), and the like. ), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), kevetlin, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. In some embodiments, the method further comprises administering to the patient an effective amount of a checkpoint inhibitor.In some of these embodiments, the checkpoint inhibitor is an antibody specific for programmed death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). In certain embodiments, the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). In certain embodiments, the CTLA-4 antibody is ipilimumab (Yervoy®). In some embodiments, the method further comprises administering to the patient an effective amount of a cancer vaccine.

[0049] In certain embodiments, the patient is premenopausal. In certain embodiments, the patient is peri-menopausal. In certain embodiments, the patient is post-menopausal.

[0050] In another aspect, a method of treating a female patient with breast cancer who is at risk for acquiring a gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene is provided, comprising administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0051] In another aspect, a method is provided for treating a female patient with breast cancer who is at risk of developing resistance to endocrine therapy. The endocrine therapy is optionally (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii), and / or (iii). The method includes administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0052] In some embodiments, the patient has primary breast cancer. In some of these embodiments, the primary breast cancer is locally advanced.

[0053] In various embodiments, the patient is being treated with endocrine therapy, optionally the endocrine therapy is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii), and / or (iii).

[0054] In another aspect, a method of treating a female patient with estrogen receptor-positive (ER+) primary breast cancer is provided, comprising administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0055] In some embodiments, the patient is at risk of developing resistance to endocrine therapy, and optionally the endocrine therapy is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii), and / or (iii).

[0056] In certain embodiments, the primary breast cancer is locally advanced.

[0057] In some embodiments, the patient is being treated with endocrine therapy, optionally wherein the endocrine therapy is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii), and / or (iii).

[0058] In another aspect, a method of treating a female patient with estrogen receptor-positive (ER+) locally advanced or metastatic breast cancer is provided, comprising administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0059] In various embodiments, the selected patient has been previously treated with one or more lines of endocrine therapy, hi certain embodiments, the patient has been previously treated with multiple lines of endocrine therapy.

[0060] In some embodiments, the endocrine therapy the patient has previously been treated with is a selective ER modulator (SERM). In certain embodiments, the SERM is tamoxifen, raloxifene, bazedoxifene, toremifene, or ospemifene.

[0061] In some embodiments, the endocrine therapy the patient has previously been treated with is a selective ER degrading drug (SERD). In certain embodiments, the SERD is fulvestrant, RAD1901, ARN-810 (GDC-0810), or AZD9496.

[0062] In some embodiments, the endocrine therapy the patient was previously treated with is an aromatase inhibitor. In certain embodiments, the aromatase inhibitor is exemestane (Aromasin®), letrozole (Femara®), or anastrozole (Arimidex®).

[0063] In some embodiments, the patient has disease progression after endocrine therapy. In some embodiments, the patient is resistant to endocrine therapy.

[0064] In various embodiments, the patient's cancer has at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. In some embodiments, the patient has previously been determined to have at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. In certain embodiments, the method further comprises the preceding step of determining that the patient has at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene.

[0065] In some embodiments, the at least one gain-of-function missense mutation is at any one of amino acids D538, Y537, L536, P535, V534, S463, V392, or E380.

[0066] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid D538. In some preferred embodiments, the mutation is D538G.

[0067] In certain embodiments, at least one gain-of-function missense mutation is at amino acid Y537. In some embodiments, the mutation is Y537S, Y537N, Y537C, or Y537Q. In some preferred embodiments, the mutation is Y537C.

[0068] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid L536. In some embodiments, the mutation is L536R or L536Q.

[0069] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid P535. In some embodiments, the mutation is P535H.

[0070] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid V534. In some embodiments, the mutation is V534E.

[0071] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid S463. In some embodiments, the mutation is S463P.

[0072] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid V392. In some embodiments, the mutation is V392I.

[0073] In certain embodiments, the at least one gain-of-function missense mutation is at amino acid E380. In some embodiments, the mutation is E380Q.

[0074] In various embodiments, lasofoxifene is administered in selected ER + It is administered as lasofoxifene tartrate to patients with locally advanced or metastatic breast cancer. In various embodiments, lasofoxifene is administered orally, intravenously, transdermally, topically, or via vaginal ring administration. In certain embodiments, lasofoxifene is administered orally. In some of these embodiments, lasofoxifene is administered orally (po) at about 0.5 mg / day to about 10 mg / day. In certain embodiments, lasofoxifene is administered orally at about 0.5 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day to about 5 mg / day. In certain embodiments, lasofoxifene is administered orally at about 1 mg / day. In certain embodiments, lasofoxifene is administered orally at about 5 mg / day. In various embodiments, lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month.

[0075] In certain embodiments, the method further comprises treating the patient with at least one additional endocrine therapy. In some embodiments, the patient is treated with the additional endocrine therapy at the original dose. In some other embodiments, the patient is treated with the additional endocrine therapy at a dose higher than the original dose. In certain embodiments, the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. In certain embodiments, the additional endocrine therapy is treatment with a selective ER degrader (SERD). In certain embodiments, the additional endocrine therapy is treatment with an aromatase inhibitor.

[0076] In various embodiments, the method comprises: +The method further comprises administering to the patient with locally advanced or metastatic breast cancer an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. In certain embodiments, the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. In some embodiments, the method further comprises administering to the patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. In certain embodiments, the mTOR inhibitor is everolimus. In some embodiments, the method further comprises administering to the patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. In some embodiments, the method further comprises administering to the patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. In certain embodiments, the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). In some embodiments, the method further comprises administering to the patient an effective amount of a histone deacetylase (HDAC) inhibitor. In some of these embodiments, the HDAC inhibitor is selected from the group consisting of vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), and the like. ), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), kevetlin, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. In some embodiments, the method further comprises administering to the patient an effective amount of a checkpoint inhibitor.In some of these embodiments, the checkpoint inhibitor is an antibody specific for programmed death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). In certain embodiments, the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). In certain embodiments, the CTLA-4 antibody is ipilimumab (Yervoy®). In some embodiments, the method further comprises administering to the patient an effective amount of a cancer vaccine.

[0077] In some embodiments, the patient is premenopausal. In certain embodiments, the patient has locally advanced or metastatic ER+ / HER2- breast cancer. In some of these embodiments, the patient is progressing during the first hormone treatment, such as a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI combined with a CDK4 / 6 inhibitor, or fulvestrant combined with a CDK4 / 6 inhibitor.

[0078] In some embodiments, the patient is perimenopausal. In certain embodiments, the patient has locally advanced or metastatic ER+ / HER2- breast cancer. In some of these embodiments, the patient is progressing during the first hormone treatment, such as a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI combined with a CDK4 / 6 inhibitor, or fulvestrant combined with a CDK4 / 6 inhibitor.

[0079] In some embodiments, the patient is postmenopausal. In certain embodiments, the patient has locally advanced or metastatic ER+ / HER2- breast cancer. In some of these embodiments, the patient has progressed during the first hormonal treatment with a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI combined with a CDK4 / 6 inhibitor, or fulvestrant combined with a CDK4 / 6 inhibitor. The present invention provides, for example, the following items. (Item 1) Estrogen receptor positive (ER)+ 1.) A method of treating a female patient suffering from breast cancer, comprising: A method comprising the step of administering to a female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. (Item 2) Estrogen receptor positive (ER) + 2. The method of claim 1, further comprising the step of selecting a patient diagnosed with breast cancer for treatment. (Item 3) 3. The method of claim 1 or 2, further comprising administering to the patient an agent selected from the group consisting of an aromatase inhibitor, a CDK4 / 6 inhibitor, an mTOR inhibitor, a PI3K inhibitor, an HSP90 inhibitor, a HER2 inhibitor, and an HDAC inhibitor. (Item 4) 4. The method of any one of items 1 to 3, wherein lasofoxifene or a pharmaceutically acceptable salt or prodrug thereof is administered as adjuvant therapy. (Item 5) 5. The method of any one of items 1 to 4, wherein the patient's breast cancer is resistant to endocrine therapy. (Item 6) 6. The method of any one of items 1 to 5, wherein the patient's cancer has at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. (Item 7) 7. The method of item 6, wherein the patient has previously been determined to have at least one gain-of-function missense mutation within the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. (Item 8) 8. The method of claim 7, further comprising the preceding step of determining that the patient has at least one gain-of-function missense mutation within the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. (Item 9) 9. The method of any one of items 6 to 8, wherein the at least one gain-of-function missense mutation is at any one of amino acids D538, Y537, L536, P535, V534, S463, V392, and E380. (Item 10) 10. The method of item 9, wherein the at least one gain-of-function missense mutation is at amino acid D538. (Item 11) 11. The method of item 10, wherein the mutation is D538G. (Item 12) 10. The method of item 9, wherein the at least one gain-of-function missense mutation is at amino acid Y537. (Item 13) 13. The method of claim 12, wherein the mutation is Y537S, Y537N, Y537C, or Y537Q. (Item 14) Item 14. The method of item 13, wherein the mutation is Y537C. (Item 15) the at least one gain-of-function missense mutation is at amino acid L536. 9. The method according to claim 9. (Item 16) Item 16. The method of item 15, wherein the mutation is L536R or L536Q. (Item 17) 10. The method of item 9, wherein the at least one gain-of-function missense mutation is at amino acid P535. (Item 18) 18. The method of item 17, wherein the mutation is P535H. (Item 19) 10. The method of item 9, wherein the at least one gain-of-function missense mutation is at amino acid V534. (Item 20) 20. The method of item 19, wherein the mutation is V534E. (Item 21) 10. The method of item 9, wherein the at least one gain-of-function missense mutation is at amino acid S463. (Item 22) 22. The method of item 21, wherein the mutation is S463P. (Item 23) 10. The method of item 9, wherein the at least one gain-of-function missense mutation is at amino acid V392. (Item 24) 24. The method of item 23, wherein the mutation is V392I. (Item 25) 10. The method of item 9, wherein the at least one gain-of-function missense mutation is at amino acid E380. (Item 26) 26. The method of item 25, wherein the mutation is E380Q. (Item 27) 5. The method of any one of items 1 to 4, wherein the patient is at risk of developing resistance to endocrine therapy, and optionally, the endocrine therapy is (i) a selective ER modulator (SERM) therapy, (ii) a selective ER degrader (SERD) therapy, (iii) an aromatase inhibitor therapy, or (iv) any combination of (i), (ii) and / or (iii). (Item 28) 28. The method of item 27, wherein the patient is at risk of acquiring a gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. (Item 29) 29. The method of item 27 or 28, wherein the patient has been previously treated with one or more lines of endocrine therapy. (Item 30) 30. The method of item 29, wherein the patient has received a previous line of endocrine therapy as primary or adjuvant therapy for the breast cancer. (Item 31) 31. The method of any one of items 1 to 30, wherein the ER+ breast cancer is a primary breast cancer. (Item 32) 31. The method of any one of items 1 to 30, wherein the ER+ breast cancer is locally advanced breast cancer. (Item 33) 31. The method of any one of items 1 to 30, wherein the ER+ breast cancer is metastatic breast cancer. (Item 34) 1. A method of treating cancer other than breast cancer in a female, comprising: a) Estrogen receptor-positive (ER) cancer other than breast cancer + ) selecting for treatment a patient diagnosed with cancer and having at least one gain-of-function mutation in the estrogen receptor 1 (ESR1) gene; and b) administering to said selected patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. A method comprising:

[0080] These and other features, aspects, and advantages of the present invention will become better understood with regard to the following description and accompanying drawings. [Brief explanation of the drawings]

[0081] [Figure 1] Figures 1A and 1B show the effect of lasofoxifene on ESR1 ligand binding domain ("LBD") mutations in Caov2 ovarian cancer cells, with Figure 1A demonstrating that the mutant receptor is constitutively active and unresponsive to 17-β estradiol ("E2"), and Figure 1B demonstrating that lasofoxifene inhibits mutant receptor activity in a dose-responsive manner.

[0082] [Figure 2] Figures 2A and 2B show the effect of lasofoxifene on ESR1 LBD mutations in SKBR3 breast adenocarcinoma cells. Figure 2A demonstrates that the mutant receptor is constitutively active and unresponsive to 17-β estradiol (E2), and Figure 2B demonstrates that lasofoxifene inhibits mutant receptor activity in a dose-responsive manner.

[0083] [Figure 3]Figures 3A and 3B show the effect of lasofoxifene on ESR1 LBD mutations in stably transfected MCF7 breast cancer cells, with Figure 3A demonstrating that lasofoxifene inhibits Y537S mutant receptor activity with increasing dose titration, and Figure 3B demonstrating that lasofoxifene inhibits D538G mutant receptor activity with increasing dose titration. DETAILED DESCRIPTION OF THE INVENTION

[0084] Endocrine therapy is ER + They are often used in the treatment and prevention of breast cancer. Different types of endocrine therapy include selective ER modulators (SERMs), such as tamoxifen; selective ER degraders (SERDs), such as fulvestrant; and aromatase inhibitors (AIs). Endocrine therapy is used to treat ER + Although it significantly improves outcomes for women with breast cancer, its effectiveness is limited by intrinsic and acquired endocrine resistance. Recent studies on the mechanisms of endocrine resistance have shown that, in some cases, estrogen receptor 1 (ESR1) gene mutations result in a conformational change of the ERα protein to a constitutively active state, resulting in ligand-independent activity that is relatively resistant to tamoxifen, fulvestrant, and estrogen deprivation. See Jeselsohn et al., Clinical Cancer Research 20(7):1757-1767 (2014).

[0085] Lasofoxifene is a nonsteroidal selective estrogen receptor modulator (SERM). It has high binding affinity to the estrogen receptor and acts as a tissue-selective estrogen agonist or antagonist. In the double-blind, placebo-controlled, randomized Postmenopausal Evaluation and Risk Reduction with Lasofoxifene (PEARL) trial, lasofoxifene was found to reduce the risk of osteoporosis. See Cummings et al., The New England Journal of Medicine 326(8):686-696 (2010). In the PEARL trial, lasofoxifene was also found to reduce the risk of breast cancer in postmenopausal women with osteoporosis. See LaCroix et al., Journal of the National Cancer Institute 102(22):1706-1715 (2010). However, the efficacy of lasofoxifene as a treatment for breast cancer and its effect on endocrine-resistant cancers has not previously been determined.

[0086] Using cell lines with engineered mutations in the ESR1 gene, the inventors discovered that lasofoxifene inhibits mutant receptor activity in a dose-responsive manner at clinically achievable concentrations, and that its efficacy is not precluded by endocrine resistance. This finding demonstrates that lasofoxifene can be used to treat ER tumors, including those with ESR1 mutations. + Locally advanced or metastatic breast cancer, ER + Primary breast cancer and other ER + This has opened up new avenues for treating cancer.

[0087] 6.1. Treatment Method Thus, in a first aspect, there is provided a method of treating cancer in women, comprising administering estrogen receptor positive (ER) + ) selecting a patient diagnosed with cancer for treatment. The selected patient is treated with an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0088] 6.1.1.ER+ Patients with cancer In various embodiments, the patient is identified as having ER by immunohistochemistry (IHC) performed on a patient's cancer sample. + In some embodiments, the patient has been diagnosed with locally advanced or metastatic ER cancer. + In some embodiments, the patient has been diagnosed with breast cancer. + In some embodiments, the patient has been diagnosed with primary breast cancer. + In some of these embodiments, the patient has been diagnosed with cancer. + In some of these embodiments, the patient has been diagnosed with ovarian cancer. + He has been diagnosed with lung cancer.

[0089] In some embodiments, cells of the patient's cancer have acquired a gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene.

[0090] In some embodiments, the patient is at risk of developing resistance to endocrine therapy. In certain embodiments, the patient is at risk of developing resistance to endocrine therapy due to increased expression of estrogen receptors. In certain embodiments, the patient is at risk of developing resistance to endocrine therapy due to increased expression of coactivators of estrogen receptors. In certain embodiments, the patient is at risk of developing resistance to endocrine therapy due to increased phosphorylation levels and activity of estrogen receptors and their coactivators. In certain embodiments, the patient is at risk of developing resistance to endocrine therapy due to changes in the tumor microenvironment and other host-related factors. In some preferred embodiments, the patient is at risk of developing resistance to endocrine therapy due to mutations in the estrogen receptor 1 (ESR1) gene.

[0091] In some of these embodiments, the endocrine therapy to which the patient is at risk of developing resistance is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor therapy (AI), or (iv) any combination of (i), (ii), and / or (iii).

[0092] 6.1.2. Prior treatment with endocrine therapy In various embodiments, ER + The cancer patient has been previously treated with one or more lines of endocrine therapy. In certain embodiments, the patient has been previously treated with one line of endocrine therapy. In certain other embodiments, the patient has been previously treated with multiple lines of endocrine therapy. In some embodiments, the patient has been previously treated with two lines of endocrine therapy. In some embodiments, the patient has been previously treated with three lines of endocrine therapy. In some embodiments, the patient has been previously treated with four or more lines of endocrine therapy.

[0093] In some embodiments, the endocrine therapy that the patient has previously been treated with is a selective ER modulator (SERM). In some embodiments, the selective ER modulator is selected from tamoxifen, raloxifene, bazedoxifene, toremifene, or ospemifene. In certain embodiments, the selective ER modulator is tamoxifen.

[0094] In some embodiments, the endocrine therapy that the patient has previously been treated with is a selective ER degrading drug (SERD). In various embodiments, the selective ER degrading drug binds to the estrogen receptor and causes proteasomal degradation of the receptor. In some embodiments, the selective ER degrading drug is selected from fulvestrant, RAD1901, ARN-810 (GDC-0810), and AZD9496. In certain embodiments, the selective ER degrading drug is fulvestrant.

[0095] In some embodiments, the endocrine therapy with which the patient was previously treated is an aromatase inhibitor (AI). In various embodiments, the aromatase inhibitor blocks the production of estrogen. In some embodiments, the aromatase inhibitor is selected from exemestane (Aromasin®), letrozole (Femara®), and anastrozole (Arimidex®).

[0096] In some embodiments, the endocrine therapy that the patient has previously undergone is ovarian suppression. In certain embodiments, ovarian suppression is achieved by oophorectomy. In certain embodiments, ovarian suppression is achieved by administration of a GnRH antagonist.

[0097] In certain embodiments, the patient's cancer has recurred or progressed after a previous endocrine therapy treatment. In some embodiments, the patient's cancer has recurred or progressed after tamoxifen treatment. In some embodiments, the patient's cancer has recurred or progressed after fulvestrant treatment. In some embodiments, the patient's cancer has recurred or progressed after aromatase inhibitor treatment. In some of these embodiments, the patient's cancer has recurred or progressed after multiple lines of endocrine therapy treatment.

[0098] In some embodiments, ER + The cancer patient has not been previously treated with endocrine therapy.

[0099] In certain embodiments, the patient is resistant to endocrine therapy other than lasofoxifene. In some embodiments, the patient has endogenous endocrine resistance. In some embodiments, the patient has acquired endocrine resistance. In certain embodiments, the patient is resistant to endocrine therapy due to increased expression of the estrogen receptor. In certain embodiments, the patient is resistant to endocrine therapy due to increased expression of coactivators of the estrogen receptor. In certain embodiments, the patient is resistant to endocrine therapy due to increased phosphorylation levels and activity of the estrogen receptor and its coactivators. In certain embodiments, the patient is resistant to endocrine therapy due to changes in the tumor microenvironment and other host-related factors. In some preferred embodiments, the patient is resistant to endocrine therapy due to genetic mutations in the estrogen receptor 1 (ESR1) gene.

[0100] In various embodiments, the patient is resistant to clinical doses of one or more SERMs other than lasofoxifene. In some of these embodiments, the patient is resistant to clinical doses of tamoxifen. In various embodiments, the patient is resistant to clinical doses of one or more SERDs. In some of these embodiments, the patient is resistant to clinical doses of fulvestrant. In various embodiments, the patient is resistant to clinical doses of one or more aromatase inhibitors. In various embodiments, the patient is resistant to supra-clinical doses of one or more SERMs other than lasofoxifene. In some of these embodiments, the patient is resistant to supra-clinical doses of tamoxifen. In various embodiments, the patient is resistant to supra-clinical doses of one or more SERDs. In some of these embodiments, the patient is resistant to supra-clinical doses of fulvestrant. In various embodiments, the patient is resistant to supra-clinical doses of one or more aromatase inhibitors.

[0101] In certain embodiments, ER + The cancer patient is not documented to have endocrine resistance. In some of these embodiments, the patient is not documented to have endocrine resistance due to limitations in the detection method.

[0102] In some embodiments, lasofoxifene is administered to patients with ER after completion of cancer treatment. + Administered to a cancer patient. In some of these embodiments, lasofoxifene is administered to a patient to treat occult micrometastases.

[0103] 6.1.3. Menopausal status In some embodiments, ER + The cancer patient is premenopausal. In a specific embodiment, the patient is premenopausal and has locally advanced or metastatic ER. + In certain embodiments, the patient is premenopausal and has locally advanced or metastatic ER cancer. + Have breast cancer.

[0104] In certain embodiments, ER + The cancer patient is perimenopausal. In a specific embodiment, the patient is perimenopausal and has locally advanced or metastatic ER. + In certain embodiments, the patient is pre- or post-menopausal and has locally advanced or metastatic ER cancer. + Have breast cancer.

[0105] In an exemplary embodiment, ER + The cancer patient is postmenopausal. In a specific embodiment, the patient is postmenopausal and has locally advanced or metastatic ER. + In certain embodiments, the patient is postmenopausal and has locally advanced or metastatic ER cancer. + Have breast cancer.

[0106] In certain embodiments, lasofoxifene is used to treat locally advanced or metastatic ER + / HER2 - In certain embodiments, lasofoxifene is administered to premenopausal women with locally advanced or metastatic breast cancer that has progressed during primary hormonal treatment with a nonsteroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. + / HER2- It is given to premenopausal women with breast cancer.

[0107] In certain embodiments, lasofoxifene is used to treat locally advanced or metastatic ER + / HER2 - In certain embodiments, lasofoxifene is administered to pre- and post-menopausal women with breast cancer who have locally advanced or metastatic ER that has progressed during primary hormonal treatment with a nonsteroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. + / HER2 - It is given to pre- and postmenopausal women with breast cancer.

[0108] In certain embodiments, lasofoxifene is used to treat locally advanced or metastatic ER + / HER2 - In certain embodiments, lasofoxifene is administered to postmenopausal women with locally advanced or metastatic breast cancer that has progressed during primary hormonal treatment with a nonsteroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. + / HER2 - It is given to postmenopausal women with breast cancer.

[0109] Mutations in the ESR1 gene In various embodiments, the patient is +The cancer has cells that have at least one mutation in the estrogen receptor 1 (ESR1) gene, which encodes the estrogen receptor alpha (ERα) protein. In some embodiments, the mutation results in ligand-independent activity of the estrogen receptor. In some embodiments, the mutation results in enhanced ligand-stimulated activity of the estrogen receptor. In some embodiments, the mutation results in resistance to endocrine therapy. In some embodiments, the mutation promotes tumor growth. In some embodiments, the mutation enhances metastatic activity of the cancer. In some preferred embodiments, the mutation is in the ERα gene. + Enhances metastatic activity in metastatic breast cancer.

[0110] In some embodiments, the mutation arises from a rare, undetectable pre-existing clone. In some embodiments, the mutation is acquired de novo during the course of endocrine therapy treatment. In some preferred embodiments, the mutation is acquired de novo during the course of endocrine therapy treatment for breast cancer. In some embodiments, the mutation is acquired de novo after multiple lines of endocrine therapy treatment. In some embodiments, the mutation is acquired de novo after multiple lines of endocrine therapy treatment for metastatic breast cancer. In various embodiments, the mutant clone expands and becomes a more dominant clone over the course of successive lines of endocrine therapy.

[0111] In some embodiments, the mutation in the ESR1 gene is a missense point mutation. In some embodiments, the mutation in the ESR1 gene is a truncation mutation. In some embodiments, the mutation in the ESR1 gene is gene amplification. In some embodiments, the mutation in the ESR1 gene is genomic rearrangement.

[0112] In some preferred embodiments, the patient has at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the ESR1 gene. +In various embodiments, at least one of the mutations is at an amino acid selected from D538, Y537, L536, P535, V534, S463, V392, and E380 (amino acids are numbered according to the ESR1 protein of NCBI accession number NP_000116.2).

[0113] In certain embodiments, the mutation increases the stability of the agonist conformation of Helix 12 of the ERα protein. In some of these embodiments, the mutation increases the binding of the estrogen receptor to its coactivators. In some of these embodiments, the mutation results in hormone-independent activity of the estrogen receptor. In some of these embodiments, the mutation results in resistance to tamoxifen, fulvestrant, and / or aromatase inhibitors.

[0114] In certain embodiments, the mutation is at amino acid D538. In certain preferred embodiments, the mutation is D538G.

[0115] In certain embodiments, the mutation is at amino acid Y537. In some of these embodiments, the mutation is Y537S, Y537N, Y537C, or Y537Q. In certain preferred embodiments, the mutation is Y537C.

[0116] In some embodiments, the mutation is at amino acid L536. In certain embodiments, the mutation is L536R or L536Q.

[0117] In some embodiments, the mutation is at amino acid P535. In certain embodiments, the mutation is P535H.

[0118] In some embodiments, the mutation is at amino acid V534. In certain embodiments, the mutation is V534E.

[0119] In some embodiments, the mutation is at amino acid S463. In certain embodiments, the mutation is S463P.

[0120] In some embodiments, the mutation is at amino acid V392. In certain embodiments, the mutation is V392I.

[0121] In some embodiments, the mutation is at amino acid E380. In certain embodiments, the mutation is E380Q.

[0122] 6.1.4.1. Detection of ESR1 gene mutations In various embodiments, the patient has previously been determined to have at least one mutation in the ESR1 gene. Some embodiments of the methods described herein further comprise detecting a mutation in the ESR1 gene.

[0123] In some embodiments, massively parallel next-generation sequencing (NGS) is used to detect estrogen receptor mutations in a patient's cancer. In certain embodiments, the entire genome is sequenced. In certain embodiments, a selected gene panel of cancer-related genes is sequenced. In certain embodiments, all coding exons within a given set of genes are sequenced. In certain embodiments, known "hotspot" regions within a given set of genes are sequenced. However, the inherent error rate of current next-generation sequencing technology is up to 1%, limiting the sensitivity and specificity of detection. In some embodiments, targeted sequencing is used to detect the presence of ESR1 mutations. Targeted sequencing allows for deeper sequencing, but is also currently limited by an error rate of 1%. In some embodiments, methods that reduce sequencing error rates are used. In certain embodiments, the Safe-Sequencing System (Safe-SeqS) is used, which tags each template molecule and enables reliable identification of rare variants. See Kinde et al., Proceedings of the National Academy of Sciences, Vol. 108 (No. 23): 9530-9535 (2011). In certain embodiments, ultrasensitive duplex sequencing is used, which independently tags and sequences each of the two strands of a DNA duplex. See Schmitt et al., Proceedings of the National Academy of Sciences, Vol. 109 (No. 36): 14508-14513 (2012). In some embodiments, digital droplet PCR is used, which emulsifies DNA into thousands to millions of droplets, encapsulating a single DNA molecule designed with a mutant-specific primer. See Vogelstein and Kinzler, Proceedings of the National Academy of Sciences, Vol. 96 (No. 16): 2322-2326 (1999) and Huggett et al., Clinical Chemistry, Vol. 61 (No. 1): 79-88 (2014).

[0124] In some embodiments, ESR1 mutation detection is performed at the time of initial diagnosis. In some embodiments, mutation detection is performed at the time of disease progression, relapse or recurrence. In some embodiments, mutation detection is performed at the time of disease progression. In some embodiments, mutation detection is performed when the disease is stable.

[0125] In some embodiments, one or more tissue specimens are obtained for mutation detection. In certain embodiments, the tissue specimen is a tumor biopsy. In certain embodiments, the tissue specimen is a tumor metastasis biopsy. In some other embodiments, a liquid biopsy is obtained for mutation detection. In certain embodiments, the liquid biopsy is circulating tumor cells (CTCs). In certain other embodiments, the liquid biopsy is cell-free DNA from a blood sample.

[0126] In specific embodiments, ESR1 mutation is monitored by circulating tumor DNA (ctDNA) analysis.In some embodiments, ctDNA analysis is carried out throughout the course of treatment.In some of these embodiments, ctDNA is extracted from patient's blood sample.In certain embodiments, ctDNA is evaluated by digital PCR analysis of ESR1 mutation.

[0127] Estradiol levels In various embodiments, patients selected for treatment based on the presence of an ESR1 gene mutation are further selected based on serum estradiol levels.

[0128] In certain embodiments, an ER having an ESR1 gene mutation +The serum estradiol level of a patient with cancer is at least 0.20 ng / dL, e.g., at least 0.25 ng / dL, at least 0.30 ng / dL, at least 0.35 ng / dL, at least 0.40 ng / dL, at least 0.45 ng / dL, at least 0.50 ng / dL, at least 0.55 ng / dL, at least 0.60 ng / dL, at least 0.65 ng / dL, at least 0.70 ng / dL, at least 0.75 ng / dL, at least 0.80 ng / dL, at least 0.85 ng / dL, at least 0.90 ng / dL, at least 0.95 ng / dL, or at least 1.0 ng / dL.

[0129] In certain embodiments, the serum estradiol level of a patient having an ESR1 gene mutation is about 0.20 ng / dL to about 1.0 ng / dL, for example, about 0.20 ng / dL to about 0.25 ng / dL, about 0.25 ng / dL to about 0.30 ng / dL, about 0.30 ng / dL to about 0.35 ng / dL, about 0.35 ng / dL to about 0.40 ng / dL, about 0.40 ng / dL to about 0.45 ng / dL, about 0.45 ng / dL to about 0.50 ng / dL, or about 0.50 ng / dL to about 0.50 ng / dL. About 0.55ng / dL, about 0.55ng / dL to about 0.60ng / dL, about 0.60ng / dL to about 0.65ng / dL, about 0.65ng / dL to about 0.70ng / dL, about 0.70ng / dL to about 0.75ng / dL, about 0.75ng / dL to about 0.80ng / dL, about 0.80ng / dL to about 0.85ng / dL, about 0.85ng / dL to about 0.90ng / dL, about 0.90ng / dL to about 0.95ng / dL, and about 0.95ng / dL to about 1.0ng / dL.

[0130] 6.1.6. Adjuvant Treatment In various embodiments, lasofoxifene is administered to patients as an adjuvant treatment. In certain embodiments, lasofoxifene is administered to patients alone as an adjuvant treatment. In certain other embodiments, lasofoxifene is administered to patients in combination with other endocrine therapies as an adjuvant treatment. In some embodiments, lasofoxifene is administered to patients after primary treatment. In some of these embodiments, lasofoxifene is administered to patients after surgical removal or debulking of cancer.

[0131] In some embodiments, lasofoxifene is administered to the patient as adjuvant therapy in combination with an aromatase inhibitor (AI). In various embodiments, the aromatase inhibitor is exemestane (Aromasin®), letrozole (Femara®), or anastrozole (Arimidex®).

[0132] In various embodiments, the aromatase inhibitor predisposes the patient to bone-related toxic effects. In some embodiments, the aromatase inhibitor predisposes the patient to osteoporosis. In some embodiments, the aromatase inhibitor predisposes the patient to osteopenia (bone loss). In some embodiments, the aromatase inhibitor predisposes the patient to fractures. In some embodiments, the aromatase inhibitor predisposes the patient to bone pain.

[0133] In various embodiments, aromatase inhibitors predispose patients to vulvovaginal atrophy (VVA).

[0134] In some embodiments, lasofoxifene is administered continuously during the administration of the aromatase inhibitor. In some other embodiments, lasofoxifene is administered cyclically during the administration of the aromatase inhibitor. In some embodiments, lasofoxifene and the aromatase inhibitor are administered together (simultaneously). In some other embodiments, lasofoxifene and the aromatase inhibitor are administered separately (sequentially).

[0135] In certain embodiments, the dosing regimen of lasofoxifene is different from the dosing regimen of the aromatase inhibitor. In some of these embodiments, the dosage of lasofoxifene is different from the dosage of the aromatase inhibitor. In some embodiments, the dosing schedule of lasofoxifene is different from the dosing schedule of the aromatase inhibitor. In some embodiments, the route of administration of lasofoxifene is different from the route of administration of the aromatase inhibitor.

[0136] In certain embodiments, the dosing regimen of lasofoxifene is the same as the dosing regimen of the aromatase inhibitor. In some embodiments, the dosage of lasofoxifene is the same as the dosage of the aromatase inhibitor. In some embodiments, the dosing schedule of lasofoxifene is the same as the dosing schedule of the aromatase inhibitor. In some embodiments, the administration route of lasofoxifene is the same as the administration route of the aromatase inhibitor.

[0137] In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for one year. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for two years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for three years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for four years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for five years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for six years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for seven years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for 8 years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for 9 years. In some embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for 10 years. In some other embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy for more than 10 years. In certain embodiments, lasofoxifene is administered to a patient in combination with an aromatase inhibitor as adjuvant therapy until the patient's cancer progresses during therapy.

[0138] In some embodiments, lasofoxifene is administered as adjuvant therapy in combination with an aromatase inhibitor to increase disease-free survival in breast cancer patients. In some embodiments, lasofoxifene is administered as adjuvant therapy in combination with an aromatase inhibitor to reduce the incidence of contralateral breast cancer. In some embodiments, lasofoxifene is administered as adjuvant therapy in combination with an aromatase inhibitor to prevent cancer recurrence or progression.

[0139] 6.2.Lasofoxifene In various embodiments, the selected patient is treated with an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. In some preferred embodiments, the lasofoxifene is administered to the selected patient as lasofoxifene tartrate.

[0140] The term "pharmaceutically acceptable salts" refers to non-toxic pharmaceutically acceptable salts. Gould, International Journal of Pharmaceutics 33:201-217 (1986) and Berge et al., Journal of See Pharmaceutical Sciences 66(1):1-19 (1977). However, other salts known to those skilled in the art can be used. Representative organic or inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, perchloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, lactic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, hydroxyethanesulfonic acid, benzenesulfonic acid, oxalic acid, pamoic acid, 2-naphthalenesulfonic acid, p-toluenesulfonic acid, cyclohexanesulfamic acid, salicylic acid, saccharinic acid, or trifluoroacetic acid. Representative organic or inorganic bases include, but are not limited to, basic or cationic salts such as benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc.

[0141] Embodiments also include prodrugs of the compounds disclosed herein. Generally, such prodrugs are functional derivatives of the compounds that can be easily converted in vivo into the required compound. Thus, in the treatment methods of the present invention, the term "administering" is intended to encompass treatment of the various disorders described by the specifically disclosed compounds, or by compounds that cannot be specifically disclosed but that are converted to the specified compound in vivo after administration to a subject. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs," H. Bundgaard, Elsevier, 1985.

[0142] Some of the crystalline forms of the compounds may exist as polymorphs and are intended to be included in the present invention. Additionally, some of the compounds may form solvates with water (i.e., hydrates) or common organic solvents, and such solvates are intended to be encompassed by some embodiments.

[0143] If the process for preparing the compounds disclosed herein results in a mixture of stereoisomers, these isomers can be separated by conventional techniques, such as preparative chromatography. The compounds can be prepared in racemic form or as individual enantiomers or diastereomers by either stereospecific synthesis or resolution. For example, compounds can be resolved into their component enantiomers or diastereomers by standard techniques, such as the formation of stereoisomeric pairs by salt formation with optically active bases, followed by fractional crystallization and regeneration of the free acid. Compounds can also be resolved by the formation of stereoisomeric esters or amides, followed by chromatographic separation and removal of the chiral auxiliary. Alternatively, compounds can be resolved using a chiral HPLC column. It should be understood that all stereoisomers, racemic mixtures, diastereomers, cis-trans isomers, and their enantiomers are encompassed in some embodiments.

[0144] Pharmaceutical Compositions Estrogen receptor positive (ER)+ ) A method for treating cancer comprises administering a therapeutically effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. The lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof of the present invention can be formulated into a pharmaceutical composition. In addition to lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof, the composition can further comprise a pharmaceutically acceptable excipient, carrier, buffer, stabilizer, or other material known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient. The precise nature of the carrier or other material may depend on the route of administration, e.g., oral, intravenous, transdermal, topical vaginal, or vaginal ring.

[0145] Pharmaceutical compositions for oral administration may be in the form of tablets, capsules, powders, or liquids. Tablets may contain solid carriers such as gelatin or adjuvants. Liquid pharmaceutical compositions usually contain liquid carriers such as water, petroleum, animal oils, vegetable oils, mineral oils, or synthetic oils. Physiological saline solution, dextrose or other sugar solutions, or glycols such as ethylene glycol, propylene glycol, or polyethylene glycol may also be included.

[0146] For parenteral administration, lasofoxifene is in the form of a parenterally acceptable aqueous solution that is pyrogen-free and has suitable pH, isotonicity, and stability. Those skilled in the art can easily prepare suitable solutions using isotonic vehicles such as sodium chloride injection, Ringer's solution, lactated Ringer's solution, etc. Preservatives, stabilizers, buffers, antioxidants, and / or other additives may be included as needed.

[0147] Pharmaceutical compositions for topical vaginal administration can be in the form of ointments, creams, gels, or lotions. Pharmaceutical compositions for topical vaginal administration often contain water, alcohol, animal oil, vegetable oil, mineral oil, or synthetic oil. Hydrocarbons (paraffin), wool fat, beeswax, macrogol, emulsifying wax, or cetrimide may also be included.

[0148] The compositions may be administered alone or in combination with other treatments, either simultaneously or sequentially, depending on the condition being treated.

[0149] 6.4 Treatment Regimen ER + In the method of administering an effective amount of lasofoxifene in the form of a pharmaceutical composition as described above for the treatment of cancer, the terms "treatment," "treating," and the like are used herein to generally mean obtaining a desired pharmacological and / or physiological effect. The effect may be preventative, in terms of completely or partially preventing a disease, condition, or its symptoms, and / or therapeutic, in terms of partially or completely curing the disease or condition and / or adverse effects caused by the disease or condition, such as symptoms. As used herein, "treatment" encompasses any treatment of a mammalian, particularly a human, disease or condition, including (a) preventing the onset of a disease or condition in a subject who may be susceptible to the disease or condition but has not yet been diagnosed as having it; (b) inhibiting a disease or condition (e.g., halting its development); or (c) alleviating a disease or condition (e.g., causing regression of the disease or condition and resulting in the improvement of one or more symptoms). Improvement of any condition can be readily assessed according to standard methods and techniques known in the art. The population of disease subjects to be treated by the method includes subjects suffering from the undesired condition or disease, as well as subjects at risk of developing the condition or disease.

[0150] The term "effective amount" means a dose that, when administered, produces the desired effect. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques. See Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999).

[0151] 6.4.1. Route of Administration In various embodiments, lasofoxifene is administered orally, intravenously, transdermally, topically, or by vaginal ring administration.

[0152] In some embodiments, lasofoxifene is administered to a patient by oral administration. In certain embodiments, lasofoxifene is administered orally at a dose of about 0.5 mg / day to about 10 mg / day, e.g., about 0.5 mg / day to about 5 mg / day, about 0.5 mg / day to about 5 mg / day, about 1 mg / day to about 5 mg / day, about 2 mg / day to about 5 mg / day, about 3 mg / day to about 5 mg / day, about 4 mg / day to about 5 mg / day, or about 0.5 mg / day to about 4 mg / day. , about 1 mg / day to about 4 mg / day orally, about 2 mg / day to about 4 mg / day orally, about 3 mg / day to about 4 mg / day orally, about 0.5 mg / day to about 3 mg / day orally, about 1 mg / day to about 3 mg / day orally, about 2 mg / day to about 3 mg / day orally, about 0.5 mg / day to about 2 mg / day orally, about 1 mg / day to about 2 mg / day orally, or about 0.5 mg / day to about 1 mg / day orally. In some embodiments, lasofoxifene is administered orally at about 0.5 mg / day. In some embodiments, lasofoxifene is administered orally at about 1 mg / day. In some embodiments, lasofoxifene is administered orally at about 1.5 mg / day. In some embodiments, lasofoxifene is administered orally at about 2 mg / day. In some embodiments, lasofoxifene is administered orally at about 2.5 mg / day. In some embodiments, lasofoxifene is administered orally at about 3 mg / day. In some embodiments, lasofoxifene is administered orally at about 3.5 mg / day. In some embodiments, lasofoxifene is administered orally at about 4 mg / day. In some embodiments, lasofoxifene is administered orally at about 4.5 mg / day. In some embodiments, lasofoxifene is administered orally at about 5 mg / day. In some embodiments, lasofoxifene is administered orally at about 6 mg / day. In some embodiments, lasofoxifene is administered orally at about 7 mg / day. In some embodiments, lasofoxifene is administered orally at about 8 mg / day. In some embodiments, lasofoxifene is administered orally at about 9 mg / day. In some embodiments, lasofoxifene is administered orally at about 10 mg / day.In some other embodiments, lasofoxifene is administered orally at greater than 10 mg / day.

[0153] In certain embodiments, when lasofoxifene is administered to a patient with cancer that has not acquired endocrine resistance, lasofoxifene may be administered orally at less than 0.5 mg / day to prevent endocrine resistance. In certain embodiments, when lasofoxifene is administered to a cancer patient as an adjuvant treatment, lasofoxifene may be administered orally at less than 0.5 mg / day to prevent endocrine resistance.

[0154] In certain embodiments, lasofoxifene is administered once daily. In certain embodiments, lasofoxifene is administered once every two days. In certain embodiments, lasofoxifene is administered once every three days. In certain embodiments, lasofoxifene is administered once every four days. In certain embodiments, lasofoxifene is administered once every five days. In certain embodiments, lasofoxifene is administered once every six days. In certain embodiments, lasofoxifene is administered once weekly. In certain embodiments, lasofoxifene is administered once every two weeks. In certain embodiments, lasofoxifene is administered once every three weeks. In certain embodiments, lasofoxifene is administered once monthly.

[0155] In some embodiments, lasofoxifene is administered to patients by vaginal ring administration. In some of these embodiments, lasofoxifene is administered once every two weeks. In some of these embodiments, lasofoxifene is administered once every three weeks. In some of these embodiments, lasofoxifene is administered once a month. In some of these embodiments, lasofoxifene is administered once every two months. In some of these embodiments, lasofoxifene is administered once every three months. In some of these embodiments, lasofoxifene is administered once every four months.

[0156] In some embodiments, lasofoxifene is administered for 1 year. + It is administered to a cancer patient. In some embodiments, lasofoxifene is administered to a patient for two years. In some embodiments, lasofoxifene is administered to a patient for three years. In some embodiments, lasofoxifene is administered to a patient for four years. In some embodiments, lasofoxifene is administered to a patient for five years. In some other embodiments, lasofoxifene is administered to a patient for more than five years. In certain embodiments, lasofoxifene is administered to a patient until the patient's cancer progresses during therapy.

[0157] Combination therapy In various embodiments, lasofoxifene is administered alone or in combination with other therapies. In certain embodiments, lasofoxifene is administered in combination with at least one other therapy. In some embodiments, lasofoxifene and other therapies are administered together (simultaneously). In some other embodiments, lasofoxifene and other therapies are administered at different times (sequentially).

[0158] In certain embodiments, the additional therapy the patient is treated with is endocrine therapy. In various embodiments, the patient is treated with at least one additional line of endocrine therapy. In some other embodiments, the patient is treated with one additional line of endocrine therapy. In some other embodiments, the patient is treated with multiple additional lines of endocrine therapy.

[0159] In some embodiments, the patient is treated with an additional endocrine therapy at the original dose. In some other embodiments, the patient is treated with an additional endocrine therapy at a higher dose than the original dose. In certain embodiments, the patient is treated with an additional endocrine therapy at a lower dose than the original dose.

[0160] In certain embodiments, the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. In some of these embodiments, the selective ER modulator is selected from tamoxifen, raloxifene, bazedoxifene, toremifene, and ospermifene. In certain embodiments, the selective ER modulator is tamoxifen.

[0161] In certain embodiments, the additional endocrine therapy is treatment with a selective ER degrading drug (SERD). In some of these embodiments, the selective ER degrading drug is selected from fulvestrant, RAD1901, ARN-810 (GDC-0810), and AZD9496. In certain embodiments, the selective ER degrading drug is fulvestrant.

[0162] In certain embodiments, the additional endocrine therapy is treatment with an aromatase inhibitor. In some of these embodiments, the aromatase inhibitor is selected from exemestane (Aromasin®), letrozole (Femara®), and anastrozole (Arimidex®).

[0163] In various embodiments, the additional therapy is administered to the patient an effective amount of a cell cycle inhibitor.In certain embodiments, the additional therapy is administered to the patient an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor.In some embodiments, the additional therapy is a CDK4 / 6 inhibitor selected from the group consisting of palbociclib, abemaciclib, and ribociclib.

[0164] In some embodiments, the additional therapy is administered to the patient an inhibitor of a pathway that crosstalks with and activates ER transcriptional activity.In certain embodiments, the additional therapy is a mammalian target of rapamycin (mTOR) inhibitor.In a specific embodiment, the mTOR inhibitor is everolimus.In some of these embodiments, lasofoxifene in combination with everolimus is administered to postmenopausal women with locally advanced or metastatic breast cancer that has progressed during non-steroidal AI and / or fulvestrant, either as monotherapy or in combination with a CDK4 / 6 inhibitor.In various embodiments, the additional therapy is a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor.

[0165] In various embodiments, the additional therapy is administration to the patient of an effective amount of a growth factor inhibitor. In certain embodiments, the additional therapy is a human epidermal growth factor receptor 2 (HER2) inhibitor. In some embodiments, the HER2 inhibitor is trastuzumab (Herceptin®). In some other embodiments, the HER2 inhibitor is trastuzumab emtansine (Kadcyla®).

[0166] In some embodiments, the additional therapy is administering to the patient an effective amount of a histone deacetylase (HDAC) inhibitor. In various embodiments, the HDAC inhibitor is selected from the group consisting of vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), The HDAC inhibitor is abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), kevetlin, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane.In certain embodiments, the HDAC inhibitor is entinostat (MS-275) under the condition that the patient has not been treated with a HER2 inhibitor.In certain other embodiments, the HDAC inhibitor is vorinostat (Zolinza®). In yet certain other embodiments, the HDAC inhibitor is romidepsin (Istodax®).

[0167] In some embodiments, the additional therapy is administering to the patient an effective amount of a checkpoint inhibitor. In certain embodiments, the checkpoint inhibitor is an antibody. In some of these embodiments, the checkpoint inhibitor is an antibody specific for programmed death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). In some embodiments, the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). In some embodiments, the CTLA-4 antibody is ipilimumab (Yervoy®).

[0168] In certain embodiments, the additional therapy is administering to the patient an effective amount of a cancer vaccine.

[0169] In some embodiments, the additional therapy is administering to the patient an effective amount of denosumab.

[0170] In some embodiments, the additional therapy is administering to the patient an effective amount of a serotonin-norepinephrine reuptake inhibitor (SNRI), a selective serotonin reuptake inhibitor (SSRI), or gabapentin. In certain embodiments, the SNRI is venlafaxine (Effexor®).

[0171] 6.4.3. Clinical Endpoints 6.4.3.1 Primary Clinical Endpoint In various embodiments, the method comprises: + In some embodiments, the method comprises administering lasofoxifene in an amount effective to increase disease-free survival in a cancer patient. + In some embodiments, the method comprises administering lasofoxifene in an amount effective to reduce recurrence of the cancer. + In some embodiments, the method comprises administering lasofoxifene in an amount effective to increase the time to cancer recurrence. + In some embodiments, the method comprises administering lasofoxifene in an amount effective to reduce metastasis of the cancer. + The method includes administering lasofoxifene in an amount effective to increase progression-free survival in a cancer patient.

[0172] In various embodiments, the method comprises: + In certain embodiments, the method comprises increasing disease-free survival in breast cancer patients. + In certain embodiments, the method comprises: + In certain embodiments, the method comprises increasing the time to recurrence of breast cancer. +In certain embodiments, the method reduces metastasis of breast cancer to bone. + In certain embodiments, the method reduces metastasis of breast cancer to tissues other than bone. + Increases progression-free survival in breast cancer patients.

[0173] In various embodiments, the methods include: + In some embodiments, the method increases disease-free survival in cancer patients. In some embodiments, the method reduces cancer recurrence in patients with endocrine resistance. In some embodiments, the method increases the time to cancer recurrence in patients with endocrine resistance. In some embodiments, the method reduces metastasis of cancer in patients with endocrine resistance. In some embodiments, the method reduces ER progression in patients with endocrine resistance. + Increase progression-free survival in cancer patients.

[0174] In some preferred embodiments, the methods comprise administering to a subject in need thereof an ER that has developed endocrine resistance. + The method increases disease-free survival, reduces recurrence, increases time to recurrence, reduces metastasis, and / or increases progression-free survival in patients with locally advanced or metastatic breast cancer. In certain embodiments, the breast cancer has developed endocrine resistance due to the acquisition of one or more of the ESR1 mutations discussed herein. In some embodiments, the method reduces selection pressure and reduces ER during treatment. + Preventing the expansion of endocrine-resistant clones in locally advanced or metastatic breast cancer.

[0175] 6.4.3.2. Secondary Clinical Endpoints In some embodiments, the methods are effective in preventing fractures and osteopenia in women who are concurrently treated with one or more drugs that cause or predispose to osteoporosis.

[0176] In some embodiments, the method is effective in decreasing vaginal pH, increasing vaginal lubrication, and / or improving vaginal cell maturation index in women who are concurrently treated with one or more drugs that cause or predispose to vulvovaginal atrophy (VVA).

[0177] In some embodiments, the method reduces one or more symptoms of sexual dysfunction in a woman who is being concomitantly treated with one or more drugs that cause or predispose to sexual dysfunction.

[0178] In some embodiments, the method treats hot flashes in women who are concurrently treated with one or more drugs that cause or predispose to hot flashes.

[0179] In some embodiments, the method increases one or more quality of life measures selected from joint pain, genitourinary symptoms, osteopenia, and fractures.

[0180] 6.5. Further Embodiments Further embodiments are provided in the following numbered embodiments. 1. A method of treating locally advanced or metastatic breast cancer in a female, comprising: a) Estrogen receptor positive (ER) + ) selecting for treatment a patient diagnosed with locally advanced or metastatic breast cancer; and b) administering to the selected patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. A method comprising: 2. The method of embodiment 1, wherein the patient has been previously treated with one or more lines of endocrine therapy. 3. The method of embodiment 2, wherein the patient has been previously treated with multiple lines of endocrine therapy. 4. The method of embodiment 2 or embodiment 3, wherein the endocrine therapy with which the patient has previously been treated is a selective ER modulator (SERM). 5. The method of embodiment 4, wherein the SERM is tamoxifen, raloxifene, bazedoxifene, toremifene, or ospemifene. 6. The method of embodiment 2 or embodiment 3, wherein the endocrine therapy with which the patient was previously treated is a selective ER degrading drug (SERD). 7. The method of embodiment 6, wherein the SERD is fulvestrant, RAD1901, ARN-810 (GDC-0810), or AZD9496. 8. The method of embodiment 2 or embodiment 3, wherein the endocrine therapy with which the patient has previously been treated is an aromatase inhibitor. 9. The method of embodiment 8, wherein the aromatase inhibitor is exemestane (Aromasin®), letrozole (Femara®), or anastrozole (Arimidex®). 10. The method of any one of embodiments 2 to 9, wherein the patient has disease progression after endocrine therapy. 11. The method of any one of embodiments 1 to 10, wherein the patient's locally advanced or metastatic cancer is resistant to endocrine therapy other than lasofoxifene. 12. The method of any one of embodiments 1 to 11, wherein the patient's locally advanced or metastatic cancer has at least one gain-of-function missense mutation within the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. 13. The method of embodiment 12, wherein the patient has previously been determined to have at least one gain-of-function missense mutation within the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. 14. The method of embodiment 13, further comprising the preceding step of determining that the patient has at least one gain-of-function missense mutation within the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. 15. The method of any one of embodiments 12 to 14, wherein the at least one gain-of-function missense mutation is at any one of amino acids D538, Y537, L536, P535, V534, S463, V392, and E380. 16. The method of embodiment 15, wherein the at least one gain-of-function missense mutation is at amino acid D538. 17. The method of embodiment 16, wherein the mutation is D538G. 18. The method of embodiment 15, wherein the at least one gain-of-function missense mutation is at amino acid Y537. 19. The method of embodiment 18, wherein the mutation is Y537S, Y537N, Y537C, or Y537Q. 20. The method of embodiment 19, wherein the mutation is Y537C. 21. The method of embodiment 15, wherein the at least one gain-of-function missense mutation is at amino acid L536. 22. The method of embodiment 21, wherein the mutation is L536R or L536Q. 23. The method of embodiment 15, wherein at least one gain-of-function missense mutation is at amino acid P535. 24. The method of embodiment 23, wherein the mutation is P535H. 25. The method of embodiment 15, wherein the at least one gain-of-function missense mutation is at amino acid V534. 26. The method of embodiment 25, wherein the mutation is V534E. 27. The method of embodiment 15, wherein at least one gain-of-function missense mutation is at amino acid S463. 28. The method of embodiment 27, wherein the mutation is S463P. 29. The method of embodiment 15, wherein at least one gain-of-function missense mutation is at amino acid V392. 30. The method of embodiment 29, wherein the mutation is V392I. 31. The method of embodiment 15, wherein at least one gain-of-function missense mutation is at amino acid E380. 32. The method of embodiment 31, wherein the mutation is E380Q. 33. The method of any one of embodiments 12 to 32, wherein the patient's serum estradiol level is at least 0.35 ng / dL. 34. The method of any one of embodiments 12 to 32, wherein the patient's serum estradiol level is between about 0.30 ng / dL and about 0.35 ng / dL. 35. The method of any one of embodiments 12 to 32, wherein the patient's serum estradiol level is between about 0.25 ng / dL and about 0.30 ng / dL. 36. The method of any one of embodiments 1-35, wherein lasofoxifene is administered as lasofoxifene tartrate. 37. The method of any one of embodiments 1 to 36, wherein lasofoxifene is administered by oral, intravenous, transdermal, topical vaginal, or vaginal ring administration. 38. The method of embodiment 37, wherein lasofoxifene is administered by oral administration. 39. The method of embodiment 38, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 10 mg orally per day. 40. The method of embodiment 39, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 5 mg orally per day. 41. The method of embodiment 40, wherein lasofoxifene is administered from about 1 mg orally per day to about 5 mg orally per day. 42. The method of embodiment 40, wherein lasofoxifene is administered orally at 1 mg / day. 43. The method of embodiment 40, wherein lasofoxifene is administered orally at 5 mg / day. 44. The method of any one of embodiments 1-43, wherein lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month. 45. The method of any one of embodiments 1 to 44, further comprising treating said patient with at least one additional endocrine therapy. 46. ​​The method of embodiment 45, wherein the patient is treated with additional endocrine therapy at the original dose. 47. The method of embodiment 45, wherein the patient is treated with an additional endocrine therapy at a dose higher than the original dose. 48. The method of any one of embodiments 45 to 47, wherein the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. 49. The method of any one of embodiments 45 to 47, wherein the additional endocrine therapy is treatment with a selective ER degrading drug (SERD). 50. The method of any one of embodiments 45 to 47, wherein the additional endocrine therapy is treatment with an aromatase inhibitor. 51. The method of any one of embodiments 1 to 44, further comprising administering to said patient an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. 52. The method of embodiment 51, wherein the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. 53. The method of any one of embodiments 1 to 44, further comprising administering to said patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. 54. The method of embodiment 53, wherein the mTOR inhibitor is everolimus. 55. The method of any one of embodiments 1 to 44, further comprising administering to said patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. 56. The method of any one of embodiments 1 to 44, further comprising administering to said patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. 57. The method of embodiment 56, wherein the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). 58. The method of any one of embodiments 1 to 44, further comprising administering to said patient an effective amount of a histone deacetylase (HDAC) inhibitor. 59. The HDAC inhibitor is vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), quevetrine, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. 60. The method of any one of embodiments 1 to 44, further comprising administering to said patient an effective amount of a checkpoint inhibitor. 61. The method of embodiment 60, wherein the checkpoint inhibitor is an antibody specific for programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). 62. The method of embodiment 61, wherein the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). 63. The method of embodiment 61, wherein the CTLA-4 antibody is ipilimumab (Yervoy®). 64. The method of any one of embodiments 1 to 44, further comprising administering to said patient an effective amount of a cancer vaccine. 65. The method of any one of embodiments 1 to 64, wherein the patient is premenopausal. 66. The method of embodiment 65, wherein the patient has locally advanced or metastatic ER+ / HER2- breast cancer. 67. The method of embodiment 65, wherein the patient is progressing during a first hormonal treatment while on a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. 68. The method of any one of embodiments 1 to 64, wherein the patient is perimenopausal. 69. The method of embodiment 68, wherein the patient has locally advanced or metastatic ER+ / HER2- breast cancer. 70. The method of embodiment 69, wherein the patient is progressing during a first hormonal treatment while on a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. 71. The method of any one of embodiments 1 to 64, wherein the patient is postmenopausal. 72. The method of embodiment 71, wherein the patient has locally advanced or metastatic ER+ / HER2- breast cancer. 73. The method of embodiment 72, wherein the patient is progressing during a first hormonal treatment while on a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. 74. A method for treating primary breast cancer in a female, comprising: a) Estrogen receptor positive (ER) + ) selecting a patient diagnosed with primary breast cancer for treatment; and b) administering to the selected patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. A method comprising: 75. The method of embodiment 74, wherein lasofoxifene is administered as lasofoxifene tartrate. 76. The method of embodiment 74 or embodiment 75, wherein lasofoxifene is administered orally, intravenously, transdermally, topically, or by vaginal ring administration. 77. The method of embodiment 76, wherein lasofoxifene is administered by oral administration. 78. The method of embodiment 77, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 10 mg orally per day. 79. The method of embodiment 78, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 5 mg orally per day. 80. The method of embodiment 79, wherein lasofoxifene is administered from about 1 mg orally per day to about 5 mg orally per day. 81. The method of embodiment 79, wherein lasofoxifene is administered orally at 1 mg / day. 82. The method of embodiment 79, wherein lasofoxifene is administered orally at 5 mg / day. 83. The method of any one of embodiments 74 to 82, wherein lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month. 84. The method of any one of embodiments 74 to 83, further comprising treating said patient with at least one additional endocrine therapy. 85. The method of embodiment 84, wherein the patient is treated with additional endocrine therapy at the original dose. 86. The method of embodiment 84, wherein the patient is treated with additional endocrine therapy at a dose higher than the original dose. 87. The method of any one of embodiments 84 to 86, wherein the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. 88. The method of any one of embodiments 84 to 86, wherein the additional endocrine therapy is treatment with a selective ER degrading drug (SERD). 89. The method of any one of embodiments 84 to 86, wherein the additional endocrine therapy is treatment with an aromatase inhibitor. 90. The method of any one of embodiments 74 to 83, further comprising administering to said patient an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. 91. The method of embodiment 90, wherein the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. 92. The method of any one of embodiments 74 to 83, further comprising administering to said patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. 93. The method of embodiment 92, wherein the mTOR inhibitor is everolimus. 94. The method of any one of embodiments 74 to 83, further comprising administering to said patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. 95. The method of any one of embodiments 74 to 83, further comprising administering to said patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. 96. The method of embodiment 95, wherein the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). 97. The method of any one of embodiments 74 to 83, further comprising administering to said patient an effective amount of a histone deacetylase (HDAC) inhibitor. 98. The HDAC inhibitor is vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), quevetrine, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. 99. The method of any one of embodiments 74 to 83, further comprising administering to said patient an effective amount of a checkpoint inhibitor. 100. The method of embodiment 99, wherein the checkpoint inhibitor is an antibody specific for programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). 101. The method of embodiment 100, wherein the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). 102. The method of embodiment 100, wherein the CTLA-4 antibody is ipilimumab (Yervoy®). 103. The method of any one of embodiments 74 to 83, further comprising administering to said patient an effective amount of a cancer vaccine. 104. The method of any one of embodiments 74 to 103, wherein the patient is premenopausal. 105. The method of any one of embodiments 74 to 103, wherein the patient is perimenopausal. 106. The method of any one of embodiments 74 to 103, wherein the patient is postmenopausal. 107. Estrogen receptor positive (ER) + 1.) A method for adjuvant therapy of breast cancer, comprising: ER + administering to a patient undergoing primary treatment for breast cancer an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof in combination with an aromatase inhibitor. A method comprising: 108. The method of embodiment 107, wherein lasofoxifene is administered continuously during administration of the aromatase inhibitor. 109. The method of embodiment 107, wherein lasofoxifene is administered cyclically during administration of the aromatase inhibitor. 110. The method of any one of embodiments 107-109, wherein the dosing regimen of lasofoxifene is different from the dosing regimen of the aromatase inhibitor. 111. The method of any one of embodiments 107-110, wherein lasofoxifene is administered as lasofoxifene tartrate. 112. The method of any one of embodiments 107 to 111, wherein the aromatase inhibitor is exemestane (Aromasin®), letrozole (Femara®), or anastrozole (Arimidex®). 113. The method of any one of embodiments 107-112, wherein lasofoxifene is administered by oral, intravenous, transdermal, topical vaginal, or vaginal ring administration. 114. The method of embodiment 113, wherein lasofoxifene is administered by oral administration. 115. The method of embodiment 114, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 10 mg orally per day. 116. The method of embodiment 115, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 5 mg orally per day. 117. The method of embodiment 116, wherein lasofoxifene is administered from about 1 mg orally per day to about 5 mg orally per day. 118. The method of embodiment 116, wherein lasofoxifene is administered orally at 1 mg / day. 119. The method of embodiment 116, wherein lasofoxifene is administered orally at 5 mg / day. 120. The method of any one of embodiments 107-119, wherein lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month. 121. The method of any one of embodiments 107 to 120, further comprising treating said patient with an additional endocrine therapy. 122. The method of embodiment 121, wherein the additional endocrine therapy is treatment with a selective ER degrading drug (SERD). 123. The method of any one of embodiments 107 to 120, further comprising administering to said patient an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. 124. The method of embodiment 123, wherein the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. 125. The method of any one of embodiments 107 to 120, further comprising administering to said patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. 126. The method of embodiment 125, wherein the mTOR inhibitor is everolimus. 127. The method of any one of embodiments 107 to 120, further comprising administering to said patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. 128. The method of any one of embodiments 107 to 120, further comprising administering to said patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. 129. The method of embodiment 128, wherein the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). 130. The method of any one of embodiments 107 to 120, further comprising administering to said patient an effective amount of a histone deacetylase (HDAC) inhibitor. 131. The HDAC inhibitor is vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), quevetrine, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. 132. The method of any one of embodiments 107 to 120, further comprising administering to said patient an effective amount of a checkpoint inhibitor. 133. The method of embodiment 132, wherein the checkpoint inhibitor is an antibody specific for programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). 134. The method of embodiment 133, wherein the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). 135. The method of embodiment 133, wherein the CTLA-4 antibody is ipilimumab (Yervoy®). 136. The method of any one of embodiments 107 to 120, further comprising administering to said patient an effective amount of a cancer vaccine. 137. The method of any one of embodiments 107-136, wherein lasofoxifene is administered in an amount and on a schedule sufficient to improve bone mass. 138. The method of any one of embodiments 107 to 136, wherein lasofoxifene is administered in an amount and on a schedule sufficient to ameliorate the symptoms of VVA. 139. The method of any one of embodiments 107-138, wherein the patient is premenopausal. 140. The method of any one of embodiments 107-138, wherein the patient is peri-menopausal. 141. The method of any one of embodiments 107 to 138, wherein the patient is postmenopausal. 142. A method for treating cancer other than breast cancer in women, comprising: a) Estrogen receptor-positive (ER) cancer other than breast cancer + ) selecting for treatment a patient diagnosed with cancer and having at least one gain-of-function mutation in the estrogen receptor 1 (ESR1) gene; and b) administering to the selected patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. A method comprising: 143. A patient goes to the ER. + 143. The method of embodiment 142, wherein the patient has been diagnosed with ovarian cancer. 144. A patient goes to the ER. + 143. The method of embodiment 142, wherein the patient has been diagnosed with lung cancer. 145. The method of any one of embodiments 142 to 144, wherein lasofoxifene is administered as lasofoxifene tartrate. 146. The method of any one of embodiments 142 to 145, wherein lasofoxifene is administered by oral, intravenous, transdermal, topical vaginal, or vaginal ring administration. 147. The method of embodiment 146, wherein lasofoxifene is administered by oral administration. 148. The method of embodiment 147, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 10 mg orally per day. 149. The method of embodiment 148, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 5 mg orally per day. 150. The method of embodiment 149, wherein lasofoxifene is administered from about 1 mg orally per day to about 5 mg orally per day. 151. The method of embodiment 149, wherein lasofoxifene is administered orally at 1 mg / day. 152. The method of embodiment 149, wherein lasofoxifene is administered orally at 5 mg / day. 153. The method of any one of embodiments 142-152, wherein lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month. 154. The method of any one of embodiments 142 to 153, further comprising treating said patient with at least one additional endocrine therapy. 155. The method of embodiment 154, wherein the patient is treated with additional endocrine therapy at the original dose. 156. The method of embodiment 154, wherein the patient is treated with additional endocrine therapy at a dose higher than the original dose. 157. The method of any one of embodiments 154 to 156, wherein the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. 158. The method of any one of embodiments 154 to 156, wherein the additional endocrine therapy is treatment with a selective ER degrading drug (SERD). 159. The method of any one of embodiments 154 to 156, wherein the additional endocrine therapy is treatment with an aromatase inhibitor. 160. The method of any one of embodiments 142 to 153, further comprising administering to said patient an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. 161. The method of embodiment 160, wherein the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. 162. The method of any one of embodiments 142 to 153, further comprising administering to said patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. 163. The method of embodiment 162, wherein the mTOR inhibitor is everolimus. 164. The method of any one of embodiments 142 to 153, further comprising administering to said patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. 165. The method of any one of embodiments 142 to 153, further comprising administering to said patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. 166. The method of embodiment 165, wherein the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). 167. The method of any one of embodiments 142 to 153, further comprising administering to said patient an effective amount of a histone deacetylase (HDAC) inhibitor. 168. The HDAC inhibitor is vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), quevetrine, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. 169. The method of any one of embodiments 142 to 153, further comprising administering to said patient an effective amount of a checkpoint inhibitor. 170. The method of embodiment 169, wherein the checkpoint inhibitor is an antibody specific for programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). 171. The method of embodiment 170, wherein the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). 172. The method of embodiment 170, wherein the CTLA-4 antibody is ipilimumab (Yervoy®). 173. The method of any one of embodiments 142 to 173, further comprising administering to said patient an effective amount of a cancer vaccine. 174. The method of any one of embodiments 142-173, wherein the patient is premenopausal. 175. The method of any one of embodiments 142-173, wherein the patient is peri-menopausal. 176. The method of any one of embodiments 142 to 173, wherein the patient is postmenopausal. 177. A method for treating a female patient suffering from breast cancer who is at risk of acquiring a gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene, comprising administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. 178. A method of treating a female patient with breast cancer who is at risk of developing resistance to endocrine therapy, wherein optionally the endocrine therapy is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii) and / or (iii), comprising administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. 179. The method of embodiment 177 or embodiment 178, wherein the patient has primary breast cancer. 180. The method of embodiment 179, wherein the primary breast cancer is locally advanced. 181. The method of any one of embodiments 177 to 180, wherein the patient is being treated with endocrine therapy, and optionally the endocrine therapy is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii) and / or (iii). 182. Estrogen receptor positive (ER) + 2.) A method of treating a female patient suffering from primary breast cancer, comprising administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. 183. The method of embodiment 182, wherein the patient is at risk of developing resistance to endocrine therapy, and optionally, the endocrine therapy is (i) a selective ER modulator (SERM) therapy, (ii) a selective ER degrader (SERD) therapy, (iii) an aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii) and / or (iii). 184. The method of embodiment 182 or embodiment 183, wherein the primary breast cancer is locally advanced. 185. The method of any one of embodiments 182 to 184, wherein the patient is being treated with endocrine therapy, and optionally the endocrine therapy is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor (AI) therapy, or (iv) any combination of (i), (ii) and / or (iii). 186. Estrogen receptor positive (ER) + 2.) A method of treating a female patient suffering from locally advanced or metastatic breast cancer, comprising administering to the female patient an effective amount of lasofoxifene, a pharmaceutically acceptable salt thereof, or a prodrug thereof. 187. The method of embodiment 186, wherein the patient has been previously treated with one or more lines of endocrine therapy. 188. The method of embodiment 186, wherein the patient has been previously treated with multiple lines of endocrine therapy. 189. The method of any one of embodiments 186 to 188, wherein the patient has disease progression after endocrine therapy. 190. The method of any one of embodiments 186 to 188, wherein the endocrine therapy with which the patient has previously been treated is a selective ER modulator (SERM). 191. The method of embodiment 190, wherein the SERM is tamoxifen, raloxifene, bazedoxifene, toremifene, or ospemifene. 192. The method of any one of embodiments 186 to 188, wherein the endocrine therapy with which the patient has previously been treated is a selective ER degrading drug (SERD). 193. The method of embodiment 192, wherein the SERD is fulvestrant, RAD1901, ARN-810 (GDC-0810), or AZD9496. 194. The method of any one of embodiments 186 to 188, wherein the endocrine therapy with which the patient has previously been treated is an aromatase inhibitor. 195. The method of embodiment 194, wherein the aromatase inhibitor is exemestane (Aromasin®), letrozole (Femara®), or anastrozole (Arimidex®). 196. The method of any one of embodiments 187 to 195, wherein the patient has disease progression after endocrine therapy. 197. The method of any one of embodiments 186 to 196, wherein the patient's locally advanced or metastatic cancer is resistant to endocrine therapy other than lasofoxifene. 198. The method of any one of embodiments 186 to 197, wherein the patient has cancer cells with at least one gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. 199. The method of embodiment 198, wherein the patient has previously been determined to have at least one gain-of-function missense mutation within the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. 200. The preliminary step is to determine that the patient has at least one gain-of-function missense mutation within the ligand-binding domain (LBD) of the estrogen receptor 1 (ESR1) gene. 198. The method of embodiment 197, further comprising: 201. The method of any one of embodiments 198 to 198, wherein at least one gain-of-function missense mutation is at any one of amino acids D538, Y537, L536, P535, V534, S463, V392, and E380. 202. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid D538. 203. The method of embodiment 202, wherein the mutation is D538G. 204. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid Y537. 205. The method of embodiment 204, wherein the mutation is Y537S, Y537N, Y537C, or Y537Q. 206. The method of embodiment 205, wherein the mutation is Y537C. 207. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid L536. 208. The method of embodiment 207, wherein the mutation is L536R or L536Q. 209. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid P535. 210. The method of embodiment 209, wherein the mutation is P535H. 211. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid V534. 212. The method of embodiment 211, wherein the mutation is V534E. 213. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid S463. 214. The method of embodiment 213, wherein the mutation is S463P. 215. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid V392. 216. The method of embodiment 215, wherein the mutation is V392I. 217. The method of embodiment 201, wherein at least one gain-of-function missense mutation is at amino acid E380. 218. The method of embodiment 217, wherein the mutation is E380Q. 219. The method of any one of embodiments 177 to 218, wherein lasofoxifene is administered as lasofoxifene tartrate. 220. The method of any one of embodiments 177 to 219, wherein lasofoxifene is administered by oral, intravenous, transdermal, topical vaginal, or vaginal ring administration. 221. The method of embodiment 220, wherein lasofoxifene is administered by oral administration. 222. The method of embodiment 221, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 10 mg orally per day. 223. The method of embodiment 222, wherein lasofoxifene is administered from about 0.5 mg orally per day to about 5 mg orally per day. 224. The method of embodiment 223, wherein lasofoxifene is administered from about 1 mg orally per day to about 5 mg orally per day. 225. The method of embodiment 223, wherein lasofoxifene is administered orally at 1 mg / day. 226. The method of embodiment 223, wherein lasofoxifene is administered orally at 5 mg / day. 227. The method of any one of embodiments 177 to 226, wherein lasofoxifene is administered once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, or once a month. 228. The method of any one of embodiments 177 to 227, further comprising treating said patient with at least one additional endocrine therapy. 229. The method of embodiment 228, wherein the patient is treated with additional endocrine therapy at the original dose. 230. The method of embodiment 228, wherein the patient is treated with additional endocrine therapy at a dose higher than the original dose. 231. The method of any one of embodiments 228 to 230, wherein the additional endocrine therapy is treatment with a selective ER modulator (SERM) other than lasofoxifene. 232. The method of any one of embodiments 228 to 230, wherein the additional endocrine therapy is treatment with a selective ER degrading drug (SERD). 233. The method of any one of embodiments 228 to 230, wherein the additional endocrine therapy is treatment with an aromatase inhibitor. 234. The method of any one of embodiments 177 to 227, further comprising administering to said patient an effective amount of a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor. 235. The method of embodiment 234, wherein the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib. 236. The method of any one of embodiments 177 to 227, further comprising administering to said patient an effective amount of a mammalian target of rapamycin (mTOR) inhibitor. 237. The method of embodiment 236, wherein the mTOR inhibitor is everolimus. 238. The method of any one of embodiments 177 to 227, further comprising administering to said patient an effective amount of a phosphoinositide 3-kinase (PI3K) inhibitor or a heat shock protein 90 (HSP90) inhibitor. 239. The method of any one of embodiments 177 to 227, further comprising administering to said patient an effective amount of a human epidermal growth factor receptor 2 (HER2) inhibitor. 240. The method of embodiment 239, wherein the HER2 inhibitor is trastuzumab (Herceptin®) or trastuzumab emtansine (Kadcyla®). 241. The method of any one of embodiments 177 to 227, further comprising administering to said patient an effective amount of a histone deacetylase (HDAC) inhibitor. 242. The HDAC inhibitor is vorinostat (Zolinza®), romidepsin (Istodax®), chidamide (Epidaza®), panobinostat (Farydak®), belinstat (Beleodaq®, PXD101), valproic acid (Depakote®, Depakene®, Stavzor®), mocetinostat (MGCD0103), abexinostat (PCI-24781), entinostat (MS-275), pracinostat (SB939), resminostat (4SC-201), gibinostat (ITF2357), xinostat (JNJ-26481585), quevetrine, CUDC-101, AR-42, tefinostat (CHR-2835), CHR-3996, 4SC202, CG200745, rosilinostat (ACY-1215), or sulforaphane. 243. The method of any one of embodiments 177 to 227, further comprising administering to said patient an effective amount of a checkpoint inhibitor. 244. The method of embodiment 243, wherein the checkpoint inhibitor is an antibody specific for programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), or cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). 245. The method of embodiment 244, wherein the PD-1 antibody is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). 246. The method of embodiment 244, wherein the CTLA-4 antibody is ipilimumab (Yervoy®). 247. The method of any one of embodiments 177 to 227, further comprising administering to said patient an effective amount of a cancer vaccine. 248. The method of any one of embodiments 177 to 247, wherein the patient is premenopausal. 249. The method of embodiment 248, wherein the patient has locally advanced or metastatic ER+ / HER2- breast cancer. 250. The method of embodiment 249, wherein the patient is progressing during a first hormonal treatment while on a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. 251. The method of any one of embodiments 177 to 247, wherein the patient is perimenopausal. 252. The method of embodiment 251, wherein the patient has locally advanced or metastatic ER+ / HER2- breast cancer. 253. The method of embodiment 252, wherein the patient is progressing during a first hormonal treatment while on a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. 254. The method of any one of embodiments 177 to 247, wherein the patient is postmenopausal. 255. The method of embodiment 254, wherein the patient has locally advanced or metastatic ER+ / HER2- breast cancer. 256. The method of embodiment 255, wherein the patient is progressing during a first hormonal treatment while on a non-steroidal aromatase inhibitor (AI), fulvestrant, an AI in combination with a CDK4 / 6 inhibitor, or fulvestrant in combination with a CDK4 / 6 inhibitor. [Example]

[0181] 6.6.Example Below are examples of specific embodiments for carrying out the present invention. The examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should naturally be allowed for.

[0182] The practice of the present invention will employ, unless otherwise indicated, conventional methods of molecular biology, cell biology, biochemistry, genetics, cancer biology and pharmacology, which are within the skill of the art, and such techniques are explained fully in the literature.

[0183] 6.6.1. Example 1: Efficacy of lasofoxifene on ESR1 LBD mutations 6.6.1.1. Method 6.6.1.1.1. Site-directed mutagenesis ExSite mutagenesis was performed on the pENTR2B ERα WT construct using Pfu Ultra Taq polymerase with the corresponding primers as summarized in Table 1 below. Primers were PNK phosphorylated. After PCR amplification, the product was digested with DpnI at 37°C for 1 hour, followed by ligation at 16°C overnight. The ligated product was transformed into DH5α bacterial cells and grown on kanamycin-resistant plates. pENTR clones were verified by sequencing and then exchanged into the pcDNA-DEST vector using the Gateway system (Invitrogen) for expression analysis. [Table 1]

[0184] 6.6.1.1.2. Cell culture Caov2 ovarian cancer cells were grown in RPMI-1640 medium (Gibco) supplemented with 8% fetal bovine serum (FBS), sodium pyruvate (NaPyr), and non-essential amino acids (NEAA) and passaged every 2–3 days. SKBR3 breast adenocarcinoma cells were grown in DMEM medium (Gibco) supplemented with 8% fetal bovine serum (FBS), sodium pyruvate (NaPyr), and non-essential amino acids (NEAA) and passaged every 2–3 days. One day before seeding for experiments, the medium was replaced with phenol red-free RPMI-1640 medium supplemented with 8% charcoal-stripped fetal bovine serum (CFS), NaPyr, and NEAA. The cells were then seeded into 96-well experimental plates in phenol red-free medium the day before transfection.

[0185] 6.6.1.1.3. Reporter Gene Assays Caov2 cells were cotransfected with the 7X-TK-ERE-TATA luciferase reporter gene (Nagel et al., Endocrinology 142(11):4721-4728 (2001)) and expression constructs for either the wild-type or mutant receptors using Fugene transfection reagent (Promega). SKBR3 cells were cotransfected with the 3X-TK-ERE-TATA luciferase reporter gene under the same conditions. pCMV-βgal was used as a control for transfection efficiency, and pcDNA was added at a final DNA concentration of 75 ng per triplicate group. Cells were treated with the indicated ligands for 5 hours posttransfection. Twenty-four hours after treatment, cells were lysed, luciferase and β-gal assays were performed as previously described (Norris et al., J Biol Chem 270(39):22777-22782 (1995)), and plates were read on a Fusion α-FP HT plate reader (PerkinElmer Life Sciences).

[0186] 6.6.1.2. Results An ERα expression construct was created to express one of four different ESR1 LBD mutations found in metastatic breast cancer patients: Y537S, Y537N, Y537C, and D538G. Jeselsohn et al., Nature Reviews Clinical See Oncology 12(10):573-583 (2015); Jeselsohn et al., Clinical Cancer Research 20(7):1757-1767 (2014); Robinson et al., Nature Genetics 45(12):1446-1451 (2013); Thomas and Gustafsson, Trends in Endocrinology and Metabolism 26(9):467-476 (2015); and Toy et al., Nature Genetics 45(12):1439-1445 (2013). The activity of these mutants was assessed in a reconstituted estrogen response element (ERE)-luciferase reporter assay in Caov2 ovarian cancer cells and SKBR3 breast cancer cells. Data were normalized to the "0" data point (no ligand) of the wild-type receptor. As previously reported (Jeselsohn et al., 2014; Robinson et al., 2013; Toy et al., 2013), all mutants studied exhibited substantial constitutive activity when compared to the activity of wild-type (WT) ERα in the absence of its ligand: 17-β estradiol (E2). While WT ERα responds to E2 in a dose-responsive manner, the transcriptional activity of the mutants is unresponsive to E2 activation (Figures 1A and 2A).

[0187] The ability of lasofoxifene to inhibit the transcriptional activity of ERα mutants was then assessed under the same conditions. All inhibition curves were obtained at 10 -9The experiments were performed in the presence of (1 nM) 17-β estradiol. Data were normalized to the "0" data point (no lasofoxifene) for each individual receptor. Plots include data from five independent experiments, and each value is the average of triplicates from each experiment. Notably, lasofoxifene effectively inhibited the transcriptional activity of all tested ERα LBD mutants in a dose-response manner (Figures 1B and 2B).

[0188] The transcriptional IC90 values ​​of lasofoxifene were also evaluated under the same conditions in Caov2 ovarian cancer cells and SKBR3 breast adenocarcinoma cells. See Maximov et al., Current Clinical Pharmacology 8(2):135-155 (2013). The evaluated transcriptional IC90 values ​​of lasofoxifene were compared to the Cmax of these compounds in the blood at doses used in previous clinical trials and approved in Europe. See Assessment Report for Fablyn, 2009 (EMA). Calculations included the Cmax of lasofoxifene at theoretical doses of 0.5 mg and 1 mg. To evaluate the possible clinical efficacy of lasofoxifene at higher concentrations, an additional dose (1 mg) of lasofoxifene was included. See Gardner et al., J Clin Pharmacol 46(1):52-58 (2006). Results from Caov2 ovarian cancer cells and SKBR3 breast adenocarcinoma cells are summarized in Table 2. [Table 2-1] [Table 2-2]

[0189] As expected, the wild-type receptor was most responsive to anti-estrogen treatment, while each of the mutants showed a reduced response to the inhibitory effects of lasofoxifene. Importantly, the pharmacology of each of the mutants differed, highlighting the need to match patients with the most appropriate drug. The data show that a 1 mg dose of lasofoxifene was most effective in patients whose tumors express the mutations in both ovarian and breast cancer settings.

[0190] 6.6.2. Example 2: Efficacy of lasofoxifene on ESR1 LBD mutations Y537S and D538G in stable transfectants MCF7 estrogen receptor alpha positive (ER) + We generated breast cancer cells stably expressing a doxycycline (DOX)-inducible hemagglutinin (HA)-tagged full-length ER carrying the ligand-binding domain mutations Y357S and D538G. Mutant introduction and expression were confirmed by Sanger sequencing, RNA sequencing, and Western blot analysis.

[0191] Dose-response studies were performed in complete media conditions. Cells were treated with DOX for induction of HA-tagged mutant ER or with vehicle as a control and seeded in triplicate. Then, on day 5, cell counts were performed using a Celigo instrument using Hoechst dye staining to detect nucleated viable cells and propidium iodide to quantify dead cells. Treatments were performed with vehicle and 10 -12 Starting at M, in 10x increments up to 10 -6 The study included increasing doses of lasofoxifene up to M. The efficacy of treatment is inversely proportional to cell count.

[0192] The antiestrogenic activity of lasofoxifene in the ER mutation Y537S and D538G breast cancer models identified in Example 1 was confirmed by the ability of lasofoxifene to overcome resistance and kill stably transfected cells with increasing dose titration (Figures 3A and 3B).

[0193] IC50 values ​​were calculated using PRISM and the results are summarized in Table 3. [Table 3]

[0194] The results confirmed that lasofoxifene treatment was effective against the Y537S and D538G mutations, but that the Y537S and D538G mutations required higher concentrations to overcome resistance.

[0195] 7. Equivalents and Incorporation by Reference While the present invention has been particularly shown and described with reference to preferred and various alternative embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.

[0196] All references, issued patents, and patent applications cited within the body of this specification are hereby incorporated by reference in their entirety for all purposes.

Claims

1. A composition comprising lasofoxifene or a pharmaceutically acceptable salt thereof for use in a method of treating a female patient suffering from estrogen receptor-positive (ER + ) breast cancer, wherein the patient is at risk of acquiring resistance to endocrine therapy, and the patient is at risk of acquiring a gain-of-function missense mutation in the ligand binding domain (LBD) of the estrogen receptor 1 (ESR1) gene, and where appropriate, the endocrine therapy is (i) selective ER modulator (SERM) therapy, (ii) selective ER degrader (SERD) therapy, (iii) aromatase inhibitor therapy, or (iv) any combination of (i), (ii) and / or (iii), the method comprising the step of administering to the patient an effective amount of lasofoxifene or a pharmaceutically acceptable salt thereof.

2. The composition described in claim 1, wherein the patient has previously been treated with one or more lines of endocrine therapy, and optionally, the patient has received one or more prior lines of endocrine therapy as primary or adjuvant therapy for the breast cancer, and optionally, the prior line of endocrine therapy was treatment with an aromatase inhibitor.

3. A composition described in any one of claims 1 or 2, wherein the ER + breast cancer is primary breast cancer, locally advanced cancer, or metastatic breast cancer.

4. The composition described in any one of claims 1 to 3, wherein the method further comprises the step of administering to the patient a drug selected from the group consisting of an aromatase inhibitor, a CDK4 / 6 inhibitor, an mTOR inhibitor, a PI3K inhibitor, an HSP90 inhibitor, a HER2 inhibitor, and an HDAC inhibitor.

5. The composition described in claim 4, wherein the method further comprises the step of administering a CDK4 / 6 inhibitor to the patient.

6. The composition described in claim 5, wherein the CDK4 / 6 inhibitor is palbociclib, abemaciclib, or ribociclib.

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