Combination therapy for treating cancer

Abstract

The present disclosure includes methods, pharmaceutical compositions, and kits for treating prostate cancer wherein AZD5305 and enzalutamide are administered in combination to a subject in need thereof.

Description

[0001] The present disclosure relates to methods of treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC), and castration-resistant prostate cancer (CRPC) in patients in need thereof. Background Art

[0002] Prostate cancer is the second most common cancer in men. In 2020, prostate cancer caused an estimated 375,304 deaths worldwide, making it the fifth leading cause of cancer death in men and accounting for 6.8% of all cancer deaths in men (Sung 2021).

[0003] Treatment of prostate cancer with androgen deprivation therapy (ADT), such as luteinizing hormone-releasing hormone (LHRH) analogs or orchiectomy, is often initially effective in controlling metastatic disease. However, patients inevitably progress from androgen sensitivity to a castration-resistant phenotype, which is associated with a 90% overall mortality rate (Scher 2015).

[0004] The recent approval of several new hormonal agents (NHAs) has significantly changed the treatment landscape for patients with metastatic castration-resistant prostate cancer (mCRPC), and NHAs are now considered the standard of care in both mCRPC and metastatic hormone-sensitive prostate cancer (mHSPC) (Mohler 2019, Parker 2020).

[0005] Both abiraterone acetate and enzalutamide in combination with ADT demonstrated robust improvements in progression-free survival (PFS) and overall survival (OS) and showed a significant prolongation in time to initiation of cytotoxic chemotherapy in patients with CRPC (Beer 2014, Ryan 2013).

[0006] In addition, recent data have demonstrated the benefits of NHA in patients with mHSPC. Compared with ADT alone, abiraterone acetate plus prednisone and ADT showed a significant survival benefit by further prolonging OS and delaying the initiation of chemotherapy and subsequent therapies (Fizazi 2019). Compared with placebo plus ADT, enzalutamide plus ADT significantly reduced the risk of radiographic progression or death and reduced the risk of PSA progression, initiation of new antitumor therapy, first symptomatic skeletal events, castration resistance, and pain progression (Armstrong 2019). A phase III trial in patients with mHSPC is underway to evaluate the combination of darolutamide with standard ADT (ARANOTE, NCT04736199).

[0007] The addition of olaparib (PARP1 / PARP2 inhibitor) to abiraterone acetate plus ADT has been shown to improve radiographic progression-free survival (rPFS) compared to abiraterone acetate alone in both men with mCRPC who had previously received docetaxel (Clarke 2018) and those who had not received prior lines of systemic therapy, regardless of homologous recombination repair gene mutation (HRRm) status (AstraZeneca press release, September 24, 2021).

[0008] It was expected that the combination of olaparib (PARP1 / PARP2 inhibitor) and enzalutamide would not be successful because enzalutamide is a strong CYP3A4 inducer (Gibbons 2015) and olaparib is a CYP3A4 substrate (Dirix 2016), so co-administration of enzalutamide with olaparib would significantly reduce olaparib exposure in patients in the multiple-dose setting.

[0009] While great advances have been made in the treatment of metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC), and castration-resistant prostate cancer (CRPC), including metastatic hormone-sensitive prostate cancer (mHSPC) and metastatic castration-resistant prostate cancer (mCRPC), many patients with these cancers live with incurable disease. Therefore, it is important to continue to find new treatments for patients with incurable cancers. Summary of the invention

[0010] In some embodiments, a method of treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject in need thereof is disclosed, the method comprising administering to the subject a first amount of AZD5305 or a pharmaceutically acceptable salt thereof, and a second amount of enzalutamide or a pharmaceutically acceptable salt thereof. In the method, the first amount and the second amount together constitute a therapeutically effective amount.

[0011] In some embodiments, disclosed is AZD5305 or a pharmaceutically acceptable salt thereof for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

[0012] In some embodiments, enzalutamide or a pharmaceutically acceptable salt thereof is disclosed for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the enzalutamide or a pharmaceutically acceptable salt thereof, and ii) AZD5305 or a pharmaceutically acceptable salt thereof.

[0013] In some embodiments, disclosed is the use of AZD5305 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC), wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the medicament comprising AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

[0014] In the above embodiments, the metastatic prostate cancer can be metastatic hormone-sensitive prostate cancer (mHSPC) or metastatic castration-resistant prostate cancer (mCRPC).

[0015] In some embodiments, a pharmaceutical product is disclosed comprising i) AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

[0016] In some embodiments, a kit is disclosed, comprising: a first pharmaceutical composition comprising AZD5305 or a pharmaceutically acceptable salt thereof; a second pharmaceutical composition comprising enzalutamide or a pharmaceutically acceptable salt thereof; and instructions for using the first pharmaceutical composition and the second pharmaceutical composition in combination.

[0017] The combination of AZD5305 and enzalutamide may result in fewer side effects or be more effective than current monotherapy or combination therapy. This may be due to the fact that AZD5305 is a selective PARP1 inhibitor. "Selective PARP1 inhibitor" means an inhibitor of the PARP enzyme that has a higher selectivity for PARP1 relative to other members of the PARP family (such as PARP2, PARP3, PARP5a and PARP6). In some embodiments, the selective PARP1 inhibitor is selective for PARP1 relative to PARP2. In some embodiments, the selective PARP1 inhibitor has a selectivity of greater than 5:1 for PARP1 relative to PARP2. In some embodiments, the selective PARP1 inhibitor has a selectivity of greater than 10:1 for PARP1 relative to PARP2. In some embodiments, the selective PARP1 inhibitor has a selectivity of greater than 100:1 for PARP1 relative to PARP2.

[0018] In some embodiments, a method of treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject in need thereof is disclosed, the method comprising administering to the subject a first amount of a selective PARP1 inhibitor (such as AZD5305) or a pharmaceutically acceptable salt thereof, and a second amount of enzalutamide or a pharmaceutically acceptable salt thereof. In the method, the first amount and the second amount together constitute a therapeutically effective amount.

[0019] In some embodiments, a selective PARP1 inhibitor (such as AZD5305) or a pharmaceutically acceptable salt thereof is disclosed for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the selective PARP1 inhibitor (such as AZD5305) or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

[0020] In some embodiments, enzalutamide or a pharmaceutically acceptable salt thereof is disclosed for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the enzalutamide or a pharmaceutically acceptable salt thereof, and ii) a selective PARP1 inhibitor (such as AZD5305) or a pharmaceutically acceptable salt thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 shows Efficacy and tolerability of the combination of AZD5305 and enzalutamide in an in vivo preclinical model of LNCaP

[0022] Figure 2 shows Efficacy of the combination of AZD5305 and enzalutamide in the in vivo preclinical model C901 DETAILED DESCRIPTION

[0023] The term "AZD5305" refers to the compound having the chemical name 5-{4-[(7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl]piperazin-1-yl}-N-methylpyridine-2-carboxamide and the structure shown below:

[0024]

[0025] AZD5305 is a potent and selective PARP1 inhibitor and PARP1-DNA trap with excellent in vivo efficacy. AZD5305 is highly selective for PARP1 relative to other PARP family members, has favorable secondary pharmacology and physicochemical properties and excellent pharmacokinetics in preclinical species, and has reduced effects on human bone marrow progenitor cells in vitro.

[0026] The synthesis of AZD5305 is described in Johannes 2021 and in WO 2021 / 013735, the contents of which are hereby incorporated by reference in their entirety. In some embodiments, the free base AZD5305 is administered to the subject. In some embodiments, a pharmaceutically acceptable salt of AZD5305 is administered to the subject. In some embodiments, crystalline AZD5305 or a pharmaceutically acceptable salt of AZD5305 is administered to the subject.

[0027] The term "enzalutamide" refers to the following compound, the chemical name of which is 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluoro-N-methylbenzamide, and the structure is shown below:

[0028]

[0029] Enzalutamide is a potent AR (androgen receptor) signaling inhibitor that blocks several steps in the AR signaling pathway. Enzalutamide competitively inhibits the binding of androgens to the AR, inhibits nuclear translocation of the activated receptor, and inhibits the association of activated AR with DNA, even in the presence of AR overexpression and in prostate cancer cells that are resistant to anti-androgen drugs. Enzalutamide treatment reduces the growth of prostate cancer cells and can induce cancer cell death and tumor regression. Enzalutamide is indicated for the treatment of adult men with mCRPC who are asymptomatic or mildly symptomatic after failure of ADT, for whom chemotherapy is not clinically indicated, and for the treatment of patients with mCSPC.

[0030] The synthesis of enzalutamide is described in US 7709517, the contents of which are hereby incorporated by reference in their entirety. In some embodiments, the free base enzalutamide is administered to the subject. In some embodiments, a pharmaceutically acceptable salt of enzalutamide is administered to the subject.

[0031] The term "pharmaceutical composition" includes a composition containing an active ingredient and a pharmaceutically acceptable excipient, carrier or diluent, wherein the active ingredient is AZD5305 or a pharmaceutically acceptable salt thereof, or enzalutamide or a pharmaceutically acceptable salt thereof. The term "pharmaceutically acceptable excipient, carrier or diluent" includes compounds, materials, compositions and / or dosage forms that are suitable for use in contact with tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications as determined by those skilled in the art, within the scope of sound medical judgment. In some embodiments, the pharmaceutical composition is a solid dosage form, such as a capsule, tablet, granule, powder or sachet. In some embodiments, the pharmaceutical composition is in the form of a sterile injectable solution in one or more aqueous or non-aqueous non-toxic parenterally acceptable buffer systems, diluents, solubilizers, co-solvents or carriers. The sterile injectable preparation may also be a sterile injectable aqueous or oily suspension or a suspension in a non-aqueous diluent, carrier or co-solvent, which may be formulated according to known procedures using one or more appropriate dispersants or wetting agents and suspending agents. The pharmaceutical composition can be a solution for intravenous (iv) bolus / infusion, or a lyophilized system (alone or with excipients) reconstituted with a buffer system with or without other excipients. The lyophilized freeze-dried material can be prepared from non-aqueous solvents or aqueous solvents. The dosage form can also be a concentrate for further dilution for subsequent infusion.

[0032] The terms "treat", "treating" and "treatment" include reducing or inhibiting the activity of an enzyme or protein associated with PARP-1, AR or metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, alleviating one or more symptoms of metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, or slowing or delaying the progression of metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject. The term "treatment" also includes reducing or inhibiting the growth of a tumor or the proliferation of cancerous cells in a subject.

[0033] The terms "inhibit," "inhibition," or "inhibiting" include a decrease in the baseline activity of a biological activity or process.

[0034] The term "subject" includes warm-blooded mammals, such as primates, dogs, cats, rabbits, rats and mice. In some embodiments, the subject is a primate, such as a human. In some embodiments, the subject has metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC).

[0035] The term "therapeutically effective amount" includes an amount of AZD5305 and an amount of enzalutamide that together will cause a biological or medical response in a subject, for example, a reduction or inhibition of the activity of an enzyme or protein associated with PARP1, AR or cancer; relief of symptoms of metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC); or a slowing or delay in the progression of metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC). In some embodiments, the term "therapeutically effective amount" includes an amount of AZD5305 and enzalutamide that together effectively at least partially alleviate, inhibit and / or alleviate metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, or inhibit PARP1 or AR, and / or reduce or inhibit the growth of a tumor or the proliferation of cancerous cells.

[0036] In some embodiments, a method of treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject in need thereof is disclosed, the method comprising administering to the subject a first amount of AZD5305 or a pharmaceutically acceptable salt thereof, and a second amount of enzalutamide or a pharmaceutically acceptable salt thereof. In the method, the first amount and the second amount together constitute a therapeutically effective amount.

[0037] In some embodiments, disclosed is AZD5305 or a pharmaceutically acceptable salt thereof for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

[0038] In some embodiments, enzalutamide or a pharmaceutically acceptable salt thereof is disclosed for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the enzalutamide or a pharmaceutically acceptable salt thereof, and ii) AZD5305 or a pharmaceutically acceptable salt thereof.

[0039] In some embodiments, disclosed is the use of AZD5305 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the medicament comprising AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

[0040] In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof and enzalutamide or a pharmaceutically acceptable salt thereof are administered separately, sequentially or simultaneously in a treatment cycle. In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof is administered continuously in a treatment cycle and enzalutamide or a pharmaceutically acceptable salt thereof is also administered continuously in the treatment cycle.

[0041] The term "continuously" or "continuously" refers to administration of a therapeutic agent, such as AZD5305, at regular intervals without cessation or interruption (ie no blank days). A "blank day" means a day on which no therapeutic agent is administered.

[0042] As used herein, a "cycle," "treatment cycle," or "dosing schedule" refers to a period of time of a combination therapy that is repeated on a regular schedule. For example, treatment may be administered for one week, two weeks, or three weeks, wherein AZD5305 and enzalutamide are administered in a coordinated manner. In some embodiments, the treatment cycle is from about 1 week to about 3 months. In some embodiments, the treatment cycle is from about 5 days to about 1 month. In some embodiments, the treatment cycle is from about 1 week to about 3 weeks. In some embodiments, the treatment cycle is about 1 week, about 10 days, about 2 weeks, about 3 weeks, about 4 weeks, about 2 months, or about 3 months.

[0043] In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof and enzalutamide or a pharmaceutically acceptable salt thereof are administered to a human subject in one or more treatment cycles (e.g., courses of treatment). A "course of treatment" comprises multiple treatment cycles, which can be repeated on a regular schedule, or adjusted to a gradually decreasing schedule based on the disease progression of the monitored patient. For example, at the beginning of a course of treatment (e.g., when the patient is first diagnosed), the patient's treatment cycle may have a longer treatment period and / or a shorter rest period, and as the cancer begins to ease, the rest period is extended, thereby increasing the length of a treatment cycle. Throughout the course of treatment, a technician can determine and adjust the time periods for treatment and rest in the treatment cycle, the number of treatment cycles, and the length of the course of treatment based on the patient's disease progression, treatment tolerance, and prognosis. In some embodiments, the method comprises 1 to 10 treatment cycles. In some embodiments, the method comprises 2 to 8 treatment cycles.

[0044] In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof is administered for 7 days in a 7 day pre-treatment cycle followed by 28 days in a 28 day treatment cycle, and enzalutamide or a pharmaceutically acceptable salt thereof is administered for 28 days in a 28 day treatment cycle.

[0045] In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof is in tablet dosage form. In some embodiments, AZD5305 is administered at a dose of up to about 60 mg per day (e.g., up to about 5 mg, up to about 10 mg, up to about 15 mg, up to about 20 mg, up to about 25 mg, up to about 30 mg, up to about 35 mg, up to about 40 mg, up to about 45 mg, up to about 50 mg, up to about 55 mg or up to about 60 mg AZD5305). In some embodiments, AZD5305 is administered once a day (QD). In some embodiments, AZD5305 is administered at a dose of about 10 mg QD, about 15 mg QD, about 20 mg QD, about 25 mg QD, about 30 mg QD, about 35 mg QD, about 40 mg QD, about 45 mg QD, about 50 mg QD, about 55 mg QD or about 60 mg QD.

[0046] In some further embodiments, AZD5305 is administered at a dose of up to about 140 mg per day (e.g., up to about 80 mg, up to about 90 mg, up to about 100 mg, up to about 110 mg, up to about 120 mg or up to about 140 mg AZD5305). In some further embodiments, AZD5305 is administered at a dose of about 80 mg QD, about 90 mg QD, about 100 mg QD, about 110 mg QD, about 120 mg QD or about 140 mg QD.

[0047] In some embodiments, enzalutamide or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, enzalutamide or a pharmaceutically acceptable salt thereof is in tablet dosage form. In some embodiments, enzalutamide or a pharmaceutically acceptable salt thereof is in capsule dosage form. In some embodiments, enzalutamide or a pharmaceutically acceptable salt thereof is administered orally once a day (QD) at a dose of about 160 mg. In some embodiments, a dose of 160 mg comprises four 40 mg capsules, four 40 mg tablets, or two 80 mg tablets.

[0048] In some embodiments, AZD5305 and enzalutamide are taken together on an empty stomach, without food for two hours before and one hour after taking the dose.

[0049] In some embodiments, a drug product is disclosed, comprising i) AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof. In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof, and enzalutamide or a pharmaceutically acceptable salt thereof are present in a single dosage form. In some embodiments, AZD5305 or a pharmaceutically acceptable salt thereof, and enzalutamide or a pharmaceutically acceptable salt thereof are present in separate dosage forms.

[0050] In some embodiments, a kit is disclosed, comprising: a first pharmaceutical composition comprising AZD5305 or a pharmaceutically acceptable salt thereof; a second pharmaceutical composition comprising enzalutamide or a pharmaceutically acceptable salt thereof; and instructions for using the first pharmaceutical composition and the second pharmaceutical composition in combination.

[0051] Metastatic prostate cancer is prostate cancer that has spread, or metastasized, to another part of the body.

[0052] Hormone-sensitive prostate cancer (HSPC) refers to cancer whose growth is inhibited by reduced androgen levels or by blocking androgen action.

[0053] Castration-resistant prostate cancer (CRPC) is a type of prostate cancer that continues to grow even when androgen levels in the body are extremely low or undetectable.

[0054] Metastatic hormone-sensitive prostate cancer (mHSPC) refers to prostate cancer that has spread, or metastasized, to another part of the body and whose growth is suppressed by low androgen levels or by blocking the action of androgens.

[0055] Metastatic castration-resistant prostate cancer (mCRPC) is prostate cancer that has spread, or metastasized, to another part of the body and continues to grow even when androgen levels in the body are very low or undetectable.

[0056] In some embodiments, luteinizing hormone releasing hormone (LHRH) agonist or antagonist therapy may be administered concurrently, especially if the patient has not undergone orchiectomy or subcapsular orchiectomy. LHRH agonists include leuprolide / leuprolide, goserelin, triptorelin, histrelin and buserelin. LHRH antagonists include degarelix, reglioxan, bicalutamide, flutamide and cyproterone acetate. Such additional treatments may be administered as per current standard of care.

[0057] Without wishing to be bound by theory, the combination of AZD5305 and enzalutamide may be beneficial because PARP1, in addition to its role in DNA repair, is a positive co-regulator of AR-driven gene expression of AR targets. (Schiewer 2012; Schiewer and Knudsen 2014). Therefore, AZD5305 should further inactivate the androgen receptor pathway, thereby enhancing the effects of enzalutamide.

[0058] In addition, new hormone agents (NHA) have been shown to induce HRR-deficient phenotypes by inhibiting AR signaling (Asim 2017; Goodwin 2013; Li 2017; Polkinghorn 2013; Tarish 2015). Homologous recombination repair gene transcripts and protein levels were found to be upregulated in prostate cancer in response to enhanced AR signaling, and increased radioresistance was observed in the presence of functional AR signaling, while reduced HRR gene expression was seen in NHA-treated cells and tumor biopsies. Therefore, without wishing to be bound by theory, the induction of the HRR-deficient phenotype by NHA will lead to increased sensitivity to AZD5305 (a selective PARP-1 inhibitor).

[0059] In some embodiments, the treated prostate cancer may lack homologous recombination (HR)-dependent DNA DSB repair activity. The HR-dependent DNA DSB repair pathway repairs double-strand breaks (DSBs) in DNA via homologous mechanisms to re-form a continuous DNA helix (Khanna and Jackson 2001). Components of the HR-dependent DNA DSB repair pathway include, but are not limited to, ATM (NM_000051), RAD51 (NM_002875), RAD51L1 (NM_002877), RAD51C (NM_002876), RAD51L3 (NM_002878), DMC1 (NM_007068), XRCC2 (NM_005431), XRCC3 (NM_005432), RAD52 (NM_002879), RAD54L (NM_003579), RAD54B (NM_012415), BRCA1 (NM_007295), BRCA2 (NM_000059), RAD50 (NM_005732), MRE11A (NM_005590) and NBS1 (NM_002485). Other proteins involved in the HR-dependent DNA DSB repair pathway include regulatory factors such as EMSY (Hughes-Davies 2003). HR components are also described in Wood 2001.

[0060] Prostate cancer defective in HR-dependent DNA DSB repair may comprise, or consist of, one or more cancer cells having reduced or abolished ability to repair DNA DSB via this pathway relative to normal cells, i.e., the activity of the HR-dependent DNA DSB repair pathway may be reduced or abolished in one or more cancer cells.

[0061] The activity of one or more components of the HR-dependent DNA DSB repair pathway can be abolished in one or more prostate cancer cells of an individual suffering from prostate cancer deficient in HR-dependent DNA DSB repair. Components of the HR-dependent DNA DSB repair pathway are well characterized in the art (see, e.g., Wood 2001) and include the components listed above.

[0062] In some embodiments, the prostate cancer cells may have a BRCA1 and / or BRCA2 defective phenotype, i.e., the activity of BRCA1 and / or BRCA2 is reduced or eliminated in the prostate cancer cells. The BRCA1 and / or BRCA2 of the prostate cancer cells having this phenotype may be defective, i.e., the expression and / or activity of BRCA1 and / or BRCA2 may be reduced or eliminated in the prostate cancer cells, for example, by means of mutations or polymorphisms in the encoding nucleic acid, or by means of amplification, mutations or polymorphisms of genes encoding regulatory factors (e.g., the EMSY gene encoding the BRCA2 regulatory factor) (Hughes-Davies 2003).

[0063] BRCA1 and BRCA2 are known tumor suppressors whose wild-type alleles are frequently lost in tumors of heterozygous carriers (Jasin 2002; Tutt 2002).

[0064] In some embodiments, the individual is heterozygous for one or more variations (such as mutations and polymorphisms) in BRCA1 and / or BRCA2 or their regulators. Detection of variations in BRCA1 and BRCA2 is well known in the art and is described, for example, in EP 699754, EP 705903, Neuhausen and Ostrander 1992; Chappuis and Foulkes 2002; Janatová 2003; Jancárková 2003). Determination of amplification of the BRCA2 binding factor EMSY is described in Hughes-Davies 2003.

[0065] Cancer-associated mutations and polymorphisms can be detected at the nucleic acid level by detecting the presence of variant nucleic acid sequences, or at the protein level by detecting the presence of variant (ie, mutant or allelic variant) polypeptides.

[0066] Examples

[0067] The compounds of the present application will now be further explained by reference to the following non-limiting examples.

[0068] Example 1. Efficacy of the combination of AZD5305 and enzalutamide in an in vitro assay

[0069] Cell lines The following cell lines were originally obtained from ATCC:

[0070]

[0071] Cell line identification was verified using the CellCheck assay (IDEXX Bioanalytics, Westbrook, ME, USA). All cell lines were verified to be free of viral mycoplasma contamination using the MycoSEQ assay (Thermo Fisher Scientific, Waltham, MA, USA) or the STAT-Myco assay (IDEXX Bioanalytics). All cell lines were grown in RPMI-1640 growth medium (Corning 17-105-CV), which was supplemented with 10% fetal bovine serum (FBS) or (when indicated) 10% charcoal-stripped FBS (Thermo Fisher Scientific, 12676029) and 2 mM glutamine.

[0072] Cell proliferation assay and combination benefit calculation

[0073] Cells in 384-well or 96-well plates were dosed using an Echo 555 (LabCyte, San Jose, CA, USA) or using an HP D300e digital dispenser (HP Life Science Dispensing). CellTiter-Glo was used to determine viable cell counts before and after treatment (7 days after treatment) according to the manufacturer's instructions (Promega, Madison, WI, USA; G7570).

[0074] Cell viability was determined using the SytoxGreen assay as described in Davies 2012, and AC was calculated. 50 The HSA (highest single agent) synergy score was calculated according to Bernenbaum 1989.

[0075] result

[0076]

[0077] The efficacy of monotherapy agents is expressed as M concentration; values ​​are the average of two independent experiments performed in triplicate. HSA (highest single agent) is the average of the synergy scores of three independent experiments performed in triplicate. SD indicates standard deviation error, where only one experiment was performed for the unindicated cases.

[0078] These results indicate that the combination of AZD5305 and enzalutamide showed synergy in LnCAP, C4-2, VCAP and CWR22Pc-R1-AD1 cell lines.

[0079] Example 2. Efficacy of the combination of AZD5305 and enzalutamide in ATMKO prostate cancer cells in an in vitro assay

[0080] An isogenic ATM-KO model was generated in LNCAP cells using CRISPR-Cas9 technology. Two clones showing very low or undetectable ATM protein levels were obtained: clone .1 (0.14 ATM protein level compared to control) and clone .2 (<0.01 ATM protein level compared to control).

[0081] Using the method of Example 1, it was determined that these ATM-KO cell lines were sensitive to AZD5305. 50 3 nM, compared with the parental (or control) LNCAP cell line which was insensitive to AZD5305 monotherapy. 50 Greater than 10 μM. No sensitivity could be measured in the cell lines responding to enzalutamide.

[0082]

[0083] When AZD5305 and enzalutamide were used in combination, the HSA (highest single agent) synergy score was calculated according to Bernenbaum 1989.

[0084]

[0085] A stronger synergistic effect was seen in the LNCAP-ATM-KO model compared to the LNCAP control cell line.

[0086] Example 3. Efficacy of the combination of AZD5305 and enzalutamide in the in vivo preclinical model LNCaP

[0087] LNCaP cells (1x 10 7 cells, 1:1 in Matrigel) were implanted subcutaneously into the flank of male NODS CID mice (5-8 weeks old, weighing approximately 25-30 g, provided by Charles River). 3 At 4:00, 40 mice with tumors of the most similar size were randomly assigned to treatment groups as shown in the table below.

[0088]

[0089] &- Enzalutamide was given 1 hour before the morning dose of AZD5305

[0090] Drug delivery preparations

[0091]

[0092] Research

[0093] Mice were dosed for 42 days, the dose for each animal was calculated on the day of dosing, and the dosing volume was 10 mg / kg.

[0094] Tumor measurement

[0095] Tumors will be measured 3 times per week using digital calipers. Tumor length and width will be measured and volume calculated using the following formula:

[0096] Volume = (length x width 2 ) / 2.

[0097] weight

[0098] Body weights of all mice in the study were measured and recorded three times per week; this information was used to calculate precise dosing for each animal.

[0099] result

[0100] LNCaP tumors were insensitive to once-daily (QD) enzalutamide 4 mg / kg or AZD5305 1 mg / kg (see Figure 1A ), and these agents as monotherapy caused no or minimal (4%) tumor growth inhibition (TGI). However, when combined, AZD5305 + enzalutamide caused a significant 55% TGI compared to the control vehicle-treated group. In addition, this effect was significantly superior to that observed in each monotherapy group. Statistical significance compared to vehicle or combination groups was evaluated using a one-tailed t-test (*, P≤0.05; **, P≤0.01; ***, P≤0.001; ns, not significant, P>0.05).

[0101] Both monotherapy and combination therapy were well tolerated (see Figure 1B ), and the treated mice showed minimal changes in body weight.

[0102] Example 4. Efficacy of the combination of AZD5305 and enzalutamide in the in vivo preclinical model C901

[0103] C901 patient-derived xenograft (PDX) models were implanted subcutaneously into the flank of donor mice and monitored weekly until they reached an ethical size (1000-2000mm 3). When the donor tumor reaches ethical size, it will measure approximately 20 mm 3 PDX fragments of the study were implanted into research mice (male NMRINude mice, 6-9 weeks old, provided by Janvierlabs). When tumors reached approximately 150 mm 3 At 4:00, 40 mice with tumors of the most similar size were randomly assigned to treatment groups as shown in the table below.

[0104]

[0105] &- Enzalutamide was given 1 hour before the morning dose of AZD5305

[0106] Drug delivery preparations

[0107]

[0108] Research

[0109] Mice were dosed for 21 days, the dose for a single animal was calculated on the day of dosing, and the dosing volume was 10 mg / kg.

[0110] Tumor measurement

[0111] Tumors were measured twice weekly using digital calipers. Tumor length and width were measured and volume was calculated using the following formula:

[0112] Volume = (length x width 2 )xπ / 6.

[0113] weight

[0114] Body weights of all mice in the study were measured and recorded twice weekly; this information was used to calculate precise dosing for each animal.

[0115] result

[0116] C901 tumors were sensitive to once daily dosing (QD) of either enzalutamide 60 mg / kg or AZD530 51 mg / kg, and these agents as monotherapy resulted in significant TGI (92% and 90%, respectively) compared to the control vehicle group (see Figure 2A ). In addition, when combined, AZD5305 + Enzalutamide caused additional tumor growth inhibition compared to the control vehicle-treated group, resulting in 66% regression (reg). Moreover, this effect was significantly superior to that observed in each monotherapy group. Statistical significance compared to vehicle or combination groups was evaluated using a one-tailed t-test (*, P≤0.05; **, P≤0.01; ***, P≤0.001; ns).

[0117] Both monotherapy and combination treatment were well tolerated, and treated mice showed minimal changes in body weight (see Figure 2B ).

[0118] Example 5. Clinical study of the combination of AZD5305 and enzalutamide in the treatment of mCRPC and mHSPC

[0119] Inclusion criteria

[0120] Patients must have a histologically confirmed diagnosis of metastatic prostate cancer.

[0121] Candidates for treatment with enzalutamide have documented evidence of current metastatic prostate cancer, where metastatic status is defined as at least one documented metastatic lesion on a bone scan or CT / MRI scan.

[0122] Surgical or medical castration within (≤) 28 days prior to the first dose of study treatment, with serum testosterone level ≤50 ng / dL (≤1.75 nmol / L). For patients who have not undergone bilateral orchiectomy, ongoing ADT with a GnRH agonist or antagonist must be initiated at least 2 weeks prior to enrollment and must be continued throughout the study.

[0123] The patient must have:

[0124] (a) Metastatic castration-resistant prostate cancer.

[0125] Patients with mCRPC should have documented prostate cancer progression at screening with at least one of the following as assessed by the investigator:

[0126] (i) PSA progression defined by: a minimum of 3 increases in PSA levels with an interval of ≥ 1 week between each determination. PSA values ​​at the screening visit should be ≥ 1 μg / L (1 ng / mL).

[0127] (ii) Radiographic progression of soft tissue disease according to RECIST criteria, with or without PSA progression.

[0128] (iii) Radiographic progression of bone metastases, with two or more documented new bone lesions on bone scan, with or without PSA progression.

[0129] Patients with mCRPC should be first- or second-line in the castration-resistant setting (should have received ≤1 prior line of systemic therapy). Androgen deprivation therapy does not count as a line of therapy. Prior use of docetaxel while patients were in the hormone-sensitive stage of their disease does not count as a line of therapy.

[0130] or

[0131] (b) Metastatic hormone-sensitive prostate cancer.

[0132] For patients with mHSPC, the following prior therapies were allowed:

[0133] (i) Prior treatment with estrogen, cyproterone acetate, or first-generation antiandrogens was allowed, as long as treatment was discontinued 3 weeks or 5 half-lives (whichever was shorter) before enrollment.

[0134] (ii) ADT ≤ 6 months before enrollment is permitted. Androgen deprivation therapy should continue on study.

[0135] (iii) Patients may have received disease-related radiation or surgery; this should have been completed at least 4 weeks prior to enrollment.

[0136] Adequate organ and bone marrow function (in the absence of transfusion or growth factor support within 14 days prior to enrollment), as defined by:

[0137]

[0138]

[0139] a In the presence of liver metastases and elevated ALT / AST between 2.5-5×ULN, patients were enrolled only if total bilirubin levels were <1.5×ULN.

[0140] ALT = alanine aminotransferase; AST = aspartate aminotransferase; INR = international normalized ratio; ULN = upper limit of normal.

[0141] ECOGPS (Eastern Cooperative Oncology Group performance status): 0-1, with no deterioration in the previous 2 weeks.

[0142] Life expectancy ≥ 16 weeks.

[0143] Enzalutamide and AZD5305 dose escalation

[0144] The starting dose of AZD5305 will be 60 mg once daily (QD). Enzalutamide will be administered once daily (QD) at 160 mg, starting on Day 1 of Cycle 1, with the combination of AZD5305 and enzalutamide administered concurrently; this will follow an initial Cycle 0 of AZD5305 60 mg once daily (OD) for 7 days as a run-in period.

[0145] In this study, the cycle length will be 28 days, with AZD5305 dosed once daily and enzalutamide dosed once daily at 160 mg. AZD5305 and enzalutamide will be taken on an empty stomach, with no food 2 hours and 1 hour after dosing. The 160 mg dose of enzalutamide will be taken as four 40 mg softgels.

[0146] If the starting dose of 60 mg QD of AZD5305 is tolerated, the dose can be escalated to 90 mg QD if necessary (while the dose of enzalutamide will be maintained at 160 mg QD), whereas if not tolerated, the dose of AZD5305 will be reduced to 40 mg QD.

[0147] The dose of AZD5305 can be further escalated up to no more than 140 mg QD.

[0148] The dose of AZD5305 may be tapered to 20 mg QD as tolerance leads or if such a dose is demonstrated to be efficacious.

[0149] All potential dose escalation and / or decrement levels following the initial dose (including intermediate dose levels of AZD5305 and exploration of alternative regimens) may be adjusted based on emerging safety, tolerability and / or PK data.

[0150] References

[0151] Many publications are cited above to more fully describe and disclose the present invention and the state of the art in the field to which the present invention belongs. The complete citations of these references are provided below. The full text of each reference in these references is incorporated herein.

[0152]

[0153]

[0154]

[0155]

[0156]

Claims

1. A method of treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject in need thereof, the method comprising administering to the subject a first amount of AZD5305 or a pharmaceutically acceptable salt thereof, and a second amount of enzalutamide or a pharmaceutically acceptable salt thereof, wherein the first amount and the second amount together constitute a therapeutically effective amount.

2. The method of claim 1, wherein the metastatic prostate cancer is metastatic hormone-sensitive prostate cancer (mHSPC) or metastatic castration-resistant prostate cancer (mCRPC).

3. A method according to claim 1 or claim 2 wherein AZD5305 is administered once daily.

4. A method according to claim 3 wherein AZD5305 is administered at a dose of up to about 60 mg per day.

5. A method according to claim 4 wherein AZD5305 is administered at a dose of 60 mg per day.

6. A method according to claim 4, wherein AZD5305 is administered at a dose of 20 mg per day.

7. The method according to any one of claims 1 to 6, wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered once daily.

8. The method of claim 7, wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered once daily at a dose of 160 mg.

9. A method according to any one of claims 1 to 8 wherein AZD5305 and enzalutamide are taken together on an empty stomach, without food being consumed within two hours prior to administration.

10. AZD5305, or a pharmaceutically acceptable salt thereof, for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) said AZD5305, or a pharmaceutically acceptable salt thereof, and ii) enzalutamide, or a pharmaceutically acceptable salt thereof.

11. AZD5305 or a pharmaceutically acceptable salt thereof for use according to claim 10, wherein the metastatic prostate cancer is metastatic hormone-sensitive prostate cancer (mHSPC) or metastatic castration-resistant prostate cancer (mCRPC).

12. AZD5305 or a pharmaceutically acceptable salt thereof for use according to claim 10 or claim 11 wherein AZD5305 is administered once daily.

13. AZD5305 or a pharmaceutically acceptable salt thereof for use according to claim 12, wherein AZD5305 is administered in a dose of up to about 60 mg per day.

14. AZD5305 or a pharmaceutically acceptable salt thereof for use according to claim 13, wherein AZD5305 is administered at a dose of 60 mg per day.

15. AZD5305 or a pharmaceutically acceptable salt thereof for use according to claim 13, wherein AZD5305 is administered at a dose of 20 mg per day.

16. AZD5305 or a pharmaceutically acceptable salt thereof for use according to any one of claims 10 to 15, wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered twice daily.

17. AZD5305 or a pharmaceutically acceptable salt thereof for use according to claim 16, wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered once daily at a dose of 160 mg.

18. AZD5305 or a pharmaceutically acceptable salt thereof for use according to any one of claims 10 to 17, wherein AZD5305 and enzalutamide are taken together on an empty stomach, without food for two hours prior to administration.

19. Enzalutamide or a pharmaceutically acceptable salt thereof for use in treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC) in a subject, wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the enzalutamide or a pharmaceutically acceptable salt thereof, and ii) AZD5305 or a pharmaceutically acceptable salt thereof.

20. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to claim 19, wherein the metastatic prostate cancer is metastatic hormone-sensitive prostate cancer (mHSPC) or metastatic castration-resistant prostate cancer (mCRPC).

21. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to claim 19 or claim 20, wherein AZD5305 is administered once daily.

22. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to claim 21, wherein AZD5305 is administered in a dose of up to about 60 mg per day.

23. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to claim 22, wherein AZD5305 is administered at a dose of 60 mg per day.

24. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to claim 22, wherein AZD5305 is administered at a dose of 20 mg per day.

25. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to any one of claims 19 to 24, wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered once daily.

26. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to claim 25, wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered once daily at a dose of 160 mg.

27. Enzalutamide or a pharmaceutically acceptable salt thereof for use according to any one of claims 19 to 26, wherein AZD5305 and enzalutamide are taken together on an empty stomach, without food for two hours prior to administration.

28. Use of AZD5305 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating metastatic prostate cancer, hormone-sensitive prostate cancer (HSPC) or castration-resistant prostate cancer (CRPC), wherein the treatment comprises administering to the subject separately, sequentially or simultaneously i) the medicament comprising AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

29. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to claim 28, wherein the metastatic prostate cancer is metastatic hormone-sensitive prostate cancer (mHSPC) or metastatic castration-resistant prostate cancer (mCRPC).

30. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to claim 28 or claim 29 wherein AZD5305 is administered once daily.

31. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to claim 30 wherein AZD5305 is administered at a dose of up to about 60 mg per day.

32. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to claim 31 , wherein AZD5305 is administered at a dose of 60 mg per day.

33. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to claim 31 , wherein AZD5305 is administered at a dose of 20 mg per day.

34. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 33 wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered twice daily.

35. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to claim 34, wherein enzalutamide or a pharmaceutically acceptable salt thereof is administered once daily at a dose of 160 mg.

36. The use of AZD5305 or a pharmaceutically acceptable salt thereof according to any one of claims 28 to 35, wherein AZD5305 and enzalutamide are taken together on an empty stomach, without food for two hours prior to administration.

37. A pharmaceutical product comprising i) AZD5305 or a pharmaceutically acceptable salt thereof, and ii) enzalutamide or a pharmaceutically acceptable salt thereof.

38. A kit comprising: a first pharmaceutical composition comprising AZD5305 or a pharmaceutically acceptable salt thereof; a second pharmaceutical composition comprising enzalutamide or a pharmaceutically acceptable salt thereof; and instructions for using the first pharmaceutical composition and the second pharmaceutical composition in combination.

39. A method, compound, use, pharmaceutical product or kit according to any preceding claim wherein AZD5305 is replaced by an alternative selective PARP1 inhibitor.

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