EA042117B1 - METHODS FOR DIAGNOSTICS AND TREATMENT OF RESISTANT OR SENSITIVE TO ABIRATERONACETATE-GLUCOCORTICOID THERAPY IN METASTATIC CASTRATION-RESISTANT PROSTATE CANCER - Google Patents

Description

Cross-references to related applications

This application claims priority over U.S. Provisional Application No. 62/402,196, filed September 30, 2016, the contents of which are incorporated herein by reference.

Technical field

The present document provides methods for the diagnosis and treatment of abiraterone acetate-glucocorticoid therapy-resistant and abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer in a patient and biomarkers for diagnosing acquired resistance to abiraterone acetate glucocorticoid therapy in a patient.

Prerequisites for the creation of the invention

Prostate cancer is the second most common type of cancer in men in the US. It is also one of the leading causes of male cancer death among all races and among Hispanic populations. In 2010, 196,038 men in the US were diagnosed with prostate cancer, and 28,560 men in the US died of prostate cancer. Demographic aging, the westernization of the diet and the development of diagnostic methods have led to an increase in the number of patients with prostate cancer in Japan in recent years.

The US Food and Drug Administration (FDA) has approved a number of therapeutic agents for use in patients with metastatic castration-resistant prostate cancer (mCRPC). Treatment options available include docetaxel, abiraterone acetate, cabazitaxel, enzalutamide, mitoxantrone, radium-223, and the therapeutic cancer vaccine sipuleucel-T. As a result, clinicians and patients are faced with the need to choose from a variety of possible treatment options and the sequence of application of such agents, which makes clinical decisions difficult.

Statement of the Invention

Provided herein are methods for diagnosing and treating abiraterone acetate-glucocorticoid therapy-resistant metastatic castration-resistant prostate cancer in a patient, comprising (consisting of and/or consisting essentially of) for a patient treated with abiraterone acetate and a glucocorticoid, diagnosing abiraterone acetate-glucocorticoid-resistant glucocorticoid therapy for metastatic castration-resistant prostate cancer in a patient with elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample, and treating the diagnosed patient with a therapeutic agent other than abiraterone acetate and a glucocorticoid, or using abiraterone acetate and a glucocorticoid in combination with an additional therapeutic agent.

Further provided are methods for diagnosing and treating abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer in a patient, comprising (consisting of and/or consisting essentially of) for a patient treated with abiraterone acetate and glucocorticoid: diagnosing abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer in a patient when the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample is not elevated, and treating the diagnosed patient with a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid.

In addition, methods are provided for detecting resistance or sensitivity to abiraterone acetate-glucocorticoid therapy in a patient having metastatic castration-resistant prostate cancer, comprising (consisting of and/or consisting essentially of) determining the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in in a biological sample from a patient treated with abiraterone acetate and glucocorticoid where elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof indicate acquired resistance to abiraterone acetate-glucocorticoid therapy, and no elevated levels of PSMA, ACADL, NPY, UBE2C, or any their combination indicates sensitivity to abiraterone acetate glucocorticoid therapy.

Described are methods of treating metastatic castration-resistant prostate cancer in a patient, comprising (consisting of and/or consisting essentially of) administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to the patient, wherein the patient does not have an elevated level of ACADL, NPY, UBE2C, or any their combinations.

In addition, the use of PSMA, ACADL, NPY, UBE2C, or any combination thereof is described to predict sensitivity or acquired resistance to abiraterone acetate glucocorticoid therapy in a patient with metastatic castration-resistant prostate cancer.

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In the methods and uses described, the glucocorticoid may be prednisone, prednisolone, hydrocortisone, dexamethasone, cortisone, or methylprednisolone.

Brief description of graphic materials

The summary of the invention, as well as the following detailed description, will be better understood when viewed in conjunction with the accompanying drawings. For the purpose of illustrating the described methods, the drawings provide examples of the methods; however, such methods are not limited to the specific embodiments described. Let's look at the drawings.

In FIG. 1A shows the level of expression at baseline and at the end of treatment (CL) for the PSMA gene.

In FIG. 1B shows the level of expression at baseline and in CL for full-length androgen receptor (AR) and androgen receptor (ARV) variants. The expression level (%) is shown in comparison with controls from 30 healthy subjects.

In FIG. 2A shows the level of PSMA gene expression at baseline and at the end of the study/treatment (CT/T) and its relationship with secondary endpoints.

In FIG. 2B shows the expression levels of the AR and ARV genes at baseline and in CT/L and their association with secondary endpoints. Asterisks indicate statistical significance (p<0.05).

In FIG. 3A shows the level of AR protein expression as measured by the number of circulating tumor cells (AR+ CTCs) at baseline and in CI/L.

In FIG. 3B shows the level of AR protein expression by the number of CTCs (AR-CTCs) at baseline and in CI/L. Connected lines show paired samples.

Detailed Description of Illustrative Embodiments

The described methods will be better understood with reference to the following detailed description in conjunction with the accompanying figures, which are part of the present description. It should be understood that the methods described are not limited to the specific methods described and/or given herein, and that the purpose of the terminology used herein is to describe specific embodiments by way of example only and is not intended to be limiting with respect to the claimed methods.

Unless expressly stated otherwise, any description relating to a possible mechanism or mode of action or reason for an improvement is understood to be illustrative only, and no limitation is intended herein on the methods described herein to the correctness or incorrectness of any such purported mechanism or mode of action or reason. improvements.

When a range of numerical values is specified or specified herein, a range includes its endpoints and all individual integer and fractional values that fall within it, and also includes each of its smaller ranges formed by all the various possible combinations of their endpoints. and falling within their integer and fractional values, which form subgroups of a larger group of values within the specified range, to the same extent as in the case of explicitly specifying each of such smaller ranges. When a range of numerical values is defined herein as greater than a given value, said range is nonetheless finite and limited at its upper end to some value that is possible in the context of the invention described herein. When a range of numerical values is defined herein as being less than a given value, said range is nevertheless limited at its lower end to some non-zero value.

All ranges are inclusive and combinable. When values are indicated as approximate using the preceding word approximately, it is to be understood that a particular meaning constitutes another embodiment. Reference to a particular numerical value includes at least that particular numerical value, unless the context clearly dictates otherwise.

It should be understood that certain elements of the described methods, which for clarity are described herein in the context of different embodiments, can also be used in combination in one embodiment. In contrast, it is also possible to provide certain elements of the described methods, presented for brevity in the context of one embodiment, individually or in any subcombination.

The following abbreviations are used in the present description: abiraterone acetate (AA); abiraterone acetate plus low dose prednisone (AA+R); circulating tumor cells (CTC) end of treatment (CL); end of study (CI); end of treatment/study (CL/I); metastatic castration-resistant prostate cancer (mCRPC); overall survival (OS); progression-free survival (PFS); and radiographic progression-free survival (rPFS).

As used herein, the singular forms include the plural.

In the present description and the claims, various terms are used that refer to aspects of the description. Such terms shall have their usual meaning in the field, unless

- 2 042117 indicates otherwise. Other specifically defined terms should be interpreted as defined herein.

Abiraterone acetate (AA) or abiraterone acetate in the form of ZYTIGA is a 17a-hydroxylase/C17,20-lyase (CYP17) inhibitor that blocks androgen synthesis in testicular, adrenal, and prostate tumors.

The term including is intended to include examples that are encompassed by the terms consisting essentially of and consisting of; similarly, the term consisting essentially of is intended to include examples that are covered by the term consisting of.

The terms end of treatment (ET), end of study (ET), and end of treatment/study (ET/T) are used interchangeably herein.

As used herein, the term therapeutically effective amount means an amount of a therapeutic agent that produces a specific therapeutic response in a patient.

As used herein, the term therapy/treatment means the use of a therapeutic agent to cure or alleviate a cancer, including prostate cancer.

The described methods are based on the detection of the possibility of using the level of mRNA for PSMA, ACADL, NPY, UBE2C or any combination thereof can be used to diagnose and/or treat abiraterone acetate-glucocorticoid therapy-resistant or abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer ( mCRPC) in a patient treated with abiraterone acetate and a glucocorticoid. The use of these biomarkers for the diagnosis and treatment of related prostate cancer is non-trivial and non-standard.

Methods for diagnosing abiraterone acetate-glucocorticoid therapy-resistant or abiraterone acetate-glucocorticoid therapy-responsive mCRPC cancer in a patient treated with abiraterone acetate and glucocorticoid include:

for a patient treated with abiraterone acetate and glucocorticoid, diagnosing abiraterone acetate-glucocorticoid therapy-resistant mCRPC cancer in a patient with elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample; or diagnosing an abiraterone acetate-glucocorticoid therapy-responsive cancer mCRPC in a patient when PSMA, ACADL, NPY, UBE2C, or any combination thereof in a patient biological sample is not elevated.

In some embodiments, the glucocorticoid is prednisone. In some embodiments, the glucocorticoid is prednisone. In some embodiments, the glucocorticoid is hydrocortisone. In some embodiments, the glucocorticoid is dexamethasone. In some embodiments, the implementation of the glucocorticoid is a cortisone. In some embodiments, the glucocorticoid is methylprednisolone.

In some embodiments, the methods further comprise, prior to diagnosis, determining the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, in a patient's biological sample.

In addition, methods for treating metastatic castration-resistant prostate cancer in a patient are described. In some embodiments, the methods include administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to a patient, wherein the patient does not have elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof. Thus, the methods can be used to treat patients with abiraterone acetate-glucocorticoid therapy-responsive mCRPC cancer. In some embodiments, methods of treating metastatic castration-resistant prostate cancer in a patient include administering to the patient a therapeutic agent other than abiraterone acetate and/or a glucocorticoid, or administering abiraterone acetate and a glucocorticoid in combination with an additional therapeutic agent, wherein the patient has elevated PSMA, ACADL, NPY, UBE2C or any combination thereof. Thus, the methods can be used to treat patients with abiraterone acetate-glucocorticoid therapy-resistant mCRPC cancer.

In some embodiments of the methods of treatment, the patient has previously received therapy with abiraterone acetate and a glucocorticoid. Thus, methods of treating mCRPC cancer may include administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to a patient who has previously received abiraterone acetate and glucocorticoid therapy and the patient does not have an elevated PSMA, ACADL, NPY, UBE2C, or any combination thereof.

In some embodiments of the methods of treatment, the glucocorticoid is prednisone. In some embodiments, the glucocorticoid is prednisolone. In some embodiments, the glucocorticoid is hydrocortisone. In some

- 3 042117 embodiments, the glucocorticoid is dexamethasone. In some embodiments, the implementation of the glucocorticoid is a cortisone. In some embodiments, the glucocorticoid is methylprednisolone.

In some embodiments of the treatment methods, the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C. Thus, the methods include administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to a patient, wherein the patient does not have an elevated level of PSMA, ACADL, NPY, UBE2C mRNA, or any combination thereof. The level of PSMA, ACADL, NPY, UBE2C, or any combination thereof can be compared to control. Accordingly, in some aspects, the patient does not have elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof, compared to controls. Acceptable controls include the previous level of PSMA, ACADL, NPY, UBE2C or any combination thereof from a patient or the level of PSMA, ACADL, NPY, UBE2C or any combination thereof from a patient or patient population who does not have metastatic castration-resistant prostate cancer. In some aspects, a control level (prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from the same patient or from a patient or population of patients who do not have mCRPC cancer) is obtained from a blood sample.

Methods for diagnosis and treatment can be combined to provide a method for diagnosing and treating abiraterone acetate-glucocorticoid therapy-resistant or abiraterone acetate-glucocorticoid therapy-responsive mCRPC cancer. Such methods can be applied to a patient who has previously received therapy with abiraterone acetate and a glucocorticoid. Methods for diagnosing and treating abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer in a patient include:

for a patient treated with abiraterone acetate and a glucocorticoid, diagnosing abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer in a patient when the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample is not elevated; and treating the diagnosed patient with a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid.

Methods for diagnosing and treating abiraterone acetate-glucocorticoid therapy-resistant metastatic castration-resistant prostate cancer in a patient include:

for a patient treated with abiraterone acetate and glucocorticoid, diagnosing abiraterone acetate-glucocorticoid therapy-resistant metastatic castration-resistant prostate cancer in a patient with elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample; and treating the diagnosed patient with a therapeutic agent other than abiraterone acetate and a glucocorticoid, or with abiraterone acetate and a glucocorticoid in combination with an additional therapeutic agent.

In some embodiments, methods for diagnosing and treating abiraterone acetate-glucocorticoid therapy-responsive or refractory metastatic castration-resistant prostate cancer further comprise determining the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, in a patient biological sample prior to diagnosis. Such a biological sample includes any patient-derived sample that contains or may contain one or more of PSMA, ACADL, NPY, or UBE2C. In some embodiments, the biological sample includes circulating tumor cells (CTCs). Characterization of biomarkers on CTCs allows for a non-invasive, real-time assessment of tumor cell dynamics, which may be desirable in the evaluation of mCRPC cancer, since the collection of fresh tumor biopsies is not part of the standard treatment procedure.

In some embodiments, the PSMA, ACADL, NPY, or UBE2C level is the PSMA, ACADL, NPY, or UBE2C mRNA level.

The described methods of diagnosis or methods of diagnosis and treatment may further include comparing the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof with a control prior to diagnosis. For example, in some embodiments, methods for diagnosing abiraterone acetate-glucocorticoid therapy-resistant or abiraterone acetate-glucocorticoid therapy-responsive mCRPC cancer in a patient treated with abiraterone acetate and a glucocorticoid include:

comparison of the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in a biological sample of a patient with a control, and the patient has previously received therapy with abiraterone acetate and glucocorticoid; and diagnosing abiraterone acetate-glucocorticoid therapy-resistant mCRPC cancer in a patient with an elevated level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample; or

- 4 042117 diagnosing abiraterone acetate-glucocorticoid therapy-responsive cancer mCRPC in a patient when the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample is not elevated.

The diagnostic method may further comprise, prior to diagnosis, determining the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, in a biological sample of a patient, the patient being previously treated with abiraterone acetate and a glucocorticoid.

In the described methods, patients can be diagnosed with abiraterone acetate glucocorticoid therapy-resistant mCRPC cancer when the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample is elevated. In the methods described, patients can be diagnosed with abiraterone acetate-glucocorticoid therapy-responsive mCRPC cancer when the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample is not elevated.

In some embodiments, methods for diagnosing treatment of abiraterone acetate-glucocorticoid therapy-responsive mCRPC cancer in a patient include:

comparison of the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in a biological sample of a patient with a control, and the patient has previously received therapy with abiraterone acetate and glucocorticoid;

diagnosing abiraterone acetate-glucocorticoid therapy-responsive mCRPC cancer in a patient when PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample is not elevated; and treating the diagnosed patient with a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid.

In other embodiments, methods for diagnosing and treating abiraterone acetate-glucocorticoid therapy-resistant mCRPC cancer in a patient include:

comparison of the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in a biological sample of a patient with a control, and the patient has previously received therapy with abiraterone acetate and glucocorticoid;

diagnosing abiraterone acetate-glucocorticoid therapy-resistant mCRPC cancer in a patient with elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample; and treating the diagnosed patient with a therapeutic agent other than abiraterone acetate and/or a glucocorticoid, or using abiraterone acetate and a glucocorticoid in combination with an additional therapeutic agent.

The diagnostic and treatment method may further comprise, prior to comparison, determining the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, in a biological sample of a patient, the patient being previously treated with abiraterone acetate and a glucocorticoid.

The control may include prior PSMA, ACADL, NPY, UBE2C, or any combination thereof from the patient. In some aspects, the prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from the patient prior to treatment with abiraterone acetate and a glucocorticoid. In some aspects, the prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from a patient during treatment with abiraterone acetate and a glucocorticoid.

The control may be the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, from a patient or population of patients who do not have mCRPC cancer.

Treatment of a diagnosed patient with a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid may include simultaneous or sequential administration of abiraterone acetate and a glucocorticoid. In some embodiments, abiraterone acetate and a glucocorticoid are co-administered. In some embodiments, the abiraterone acetate and the glucocorticoid are administered sequentially in any order.

Treatment of a diagnosed patient with a therapeutic agent other than abiraterone acetate and a glucocorticoid, or with abiraterone acetate and a glucocorticoid in combination with an additional therapeutic agent, may include treating the patient with a known mCRPC cancer therapy in place of or in addition to abiraterone acetate and a glucocorticoid.

In addition, methods for detecting resistance or sensitivity to abiraterone acetate-glucocorticoid therapy in a patient with mCRPC cancer are described, including:

determination of the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in a biological sample from a patient treated with abiraterone acetate and glucocorticoid, and an increased level of PSMA, ACADL, NPY, UBE2C, or any combination of them indicates acquired resistance to abiraterone acetate-glucocorticoid therapy, and however, the absence of elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination of them indicates sensitivity to abiraterone acetate-glucocorticoid therapy.

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In some embodiments, the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C.

In some embodiments of methods for detecting resistance or susceptibility, the glucocorticoid is prednisone. In some embodiments, the glucocorticoid is prednisone. In some embodiments, the glucocorticoid is hydrocortisone. In some embodiments, the glucocorticoid is dexamethasone. In some embodiments, the implementation of the glucocorticoid is a cortisone. In some embodiments, the glucocorticoid is methylprednisolone.

In some embodiments, the patient has or does not have elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof, compared to a control. The control may include prior PSMA, ACADL, NPY, UBE2C, or any combination thereof from the patient. In some aspects, the prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from the patient prior to treatment with abiraterone acetate and a glucocorticoid. In some aspects, the prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from a patient during treatment with abiraterone acetate and a glucocorticoid. The control substance may include the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, from a patient or patient population that does not have mCRPC cancer.

In addition, the use of PSMA, ACADL, NPY, UBE2C, or any combination thereof to predict sensitivity or acquired resistance to abiraterone acetate glucocorticoid therapy in a patient with mCRPC cancer is described. Additionally provided is the use of PSMA, ACADL, NPY, UBE2C, or any combination thereof, for use in predicting sensitivity or acquired resistance to abiraterone acetate-glucocorticoid therapy in a patient having metastatic castration-resistant prostate cancer.

In some embodiments of the described uses, the glucocorticoid is prednisone. In some embodiments, the glucocorticoid is prednisone. In some embodiments, the glucocorticoid is hydrocortisone. In some embodiments, the glucocorticoid is dexamethasone. In some embodiments, the implementation of the glucocorticoid is a cortisone. In some embodiments, the glucocorticoid is methylprednisolone.

In some embodiments, PSMA, ACADL, NPY, UBE2C, or any combination thereof is mRNA obtained from a biological sample from a patient and the patient has previously received abiraterone acetate and glucocorticoid therapy.

The amount of abiraterone acetate administered to a patient may be from about 250 to about 1500 mg/day, from about 500 to about 1000 mg/day, about 500 mg/day, about 750 mg/day, or about 1000 mg/day.

The amount of glucocorticoid administered to a patient may be from about 2.5 to about 15 mg/day, from about 5 to about 10 mg/day, about 5 mg/day, about 7.5 mg/day, about 10 mg/day, or about 12 .5 mg/day. For example, in some embodiments, the glucocorticoid is prednisone and the amount of prednisone administered is about 2.5 to about 15 mg/day, about 5 to about 10 mg/day, about 5 mg/day, about 7.5 mg/day. day, approximately 10 mg/day or approximately 12.5 mg/day.

In some embodiments, the glucocorticoids may be prednisolone, hydrocortisone, dexamethasone, cortisone, and methylprednisolone. The amounts of prednisolone, hydrocortisone, dexamethasone, cortisone, and methylprednisolone can be from about 2 to about 10 mg/day, from 3 to 6, 4, or 5 mg/day.

Examples

The following examples are provided to further describe some of the embodiments described herein. The examples are intended to illustrate, but not to limit the described embodiments.

Study plan and treatment procedures.

Two phase 2 studies, called JPN-201 and JPN-202, were conducted in Japan. The JPN201 and JPN-202 trials were phase 2, open-label, multicentre, uncontrolled clinical trials of 1000 mg abiraterone acetate plus 10 mg prednisone daily (AA) in Japanese patients with mCRPC who had not previously received chemotherapy (JPN-201) and were previously treated course of chemotherapy (JPN-202). Patients received oral abiraterone acetate (1000 mg once a day) daily at least 1 hour before a meal and 2 hours after a meal at any time up to 10 pm. In addition, 5 mg of prednisolone was given orally twice a day in parallel. The 28 day dosing cycle was continued until disease progression or unacceptable toxicity was detected.

The primary endpoint was the proportion of patients achieving a prostate-specific antigen (PSA) response (e.g. >50% drop in PSA from baseline)

- 6 042117 for 12 weeks of treatment in accordance with the criteria of the prostate-specific antigen working group (PSAWG). Secondary endpoints included:

the proportion of PSA response (confirmed or unconfirmed) during the treatment period;

overall survival (OS);

PSA progression-free survival (PSA-PFS) based on disease progression determination according to PSAWG criteria;

radiographic progression-free survival (rPFS) based on disease progression determination according to RECIST Version 1.0 criteria; and modified PFS.

Patients were required to have a PSA level >5 ng/mL and an Eastern United Cancer Group (ECOG PS) global health score of 0 or 1. Patients were receiving study-contemplated treatment until disease progression or unacceptable toxicity was detected, unless the investigator considered that they continued to receive clinical benefit. The study was approved by the expert councils of all participating organizations and was conducted in accordance with the Declaration of Helsinki. All patients gave written informed consent to participate in the study.

Biomarker analysis.

Based on patient data from the JPN-201 and JPN-202 studies, an exploratory biomarker analysis was performed to investigate potential differences in efficacy by comparing PSA response and survival characteristics between the two studies and to investigate the molecular mechanism of abiraterone acetate resistance in a pooled population of Japanese patients with mCRPC cancer. This analysis assessed a number of biomarkers associated with the mechanism of action of abiraterone acetate or with the development and progression of prostate cancer (Table 1).

Table 1 Biomarkers assessed

Biomarker Analysis Moment of time CSC Counting ³ Baseline, C2D1, C3D1, C4D1 and CL Expression of AR ^b CellSearch® CXC Kit ^c Baseline and CL Expression of mRNA biomarker ^a RT-PCR Baseline and CL

^a CTCs were stratified as >5 or <5 CTCs, ^b nuclear AR protein expression was dichotomized as <10% or >10% in CTCs, ^c AR protein expression level was determined using the Cell-

Search® CXC Kit, using phycoerythrin-labeled monoclonal antibody to determine the level of AR protein expression, ^d additional evaluable mRNA biomarkers, including AR FL, splicing variants

AR, KLK3 (PSA), PSMA, CYP17, IFIH1, CDH1, were dichotomized as positive or negative for expression based on thresholds established from analyzes of normal healthy patients (using peak expression levels in blood samples from normal healthy patients as cutoffs).

Baseline was defined as C1D1.

AR (androgen receptor); CTC (circulating tumor cells); C2D1 (cycle 2, day 1); CL (end of treatment).

The identification of biomarkers and their association with clinical outcomes taken at baseline and at the end of the study or progression was assessed in each study and compared between studies. Association analysis of biomarkers with clinical endpoints was initially performed for the primary endpoints of the study. When positive associations were found, additional associative analysis was performed for the secondary endpoints of the study. Molecular characteristics of a subset of patients who did not respond to abiraterone acetate therapy were assessed and compared with those of patients who received a good response to treatment to identify biomarker profiles that correlate with resistance to abiraterone acetate therapy.

Samples of biomaterials were taken from patients at various time points indicated in Table. 1. Blood samples (10 ml) were taken to count the number of CTCs on day 1 from cycle 1 (baseline) to cycle 4 and at the end of treatment (CL). Additional blood samples to determine the level of AR protein expression were taken on day 1 of cycle 1 (baseline) and at the end of treatment (CL). For RNA analysis, blood samples were taken with EDTA at baseline and in CL.

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Calculation of the number of CTCs.

CTC counts were performed at the Clinical Research Solutions (CRS) Laboratory (Janssen Diagnostics, LLC, Huntington Valley, PA, USA). CTCs were quantified using the CellSearch® CTC Kit (part number 7900001) following the manufacturer's instructions. Samples were processed on a CellTracks® AutoPrep® system and analyzed on a CellTracks® Analyzer II.

AR protein expression.

AR protein expression level analysis (CTC nuclear staining) was performed in the CRS laboratory using the CellSearch® CXC Kit (p/n 7900017) following the manufacturer's instructions. To determine the level of AR protein expression, CTCs were stained using a phycoerythrin-labeled monoclonal antibody. To obtain a background signal for the phycoerythrin channel and a control substance for the reagent, respectively, normal blood samples with injected LNCaP cells were treated without the marker reagent for the AR protein (background control) and in the presence of the marker reagent for the AR protein (positive control).

Analysis of gene expression.

Using a quantitative reverse transcriptase polymerase chain reaction (RT-PCR), a gene expression analysis panel was developed for mRNA biomarkers from specific RNA groups. Biomarker analysis was performed as a single marker analysis. We developed assays to detect full-length ARs (AR FL) and AR splicing variants lacking a ligand-binding domain (ARV1, ARV7 and ARV567), GAPDH and RPL19 (universal controls), PSMA, CDH1, IFIH1, NPY, UBE2C and ACADL using cell-based line model of the prostate (LNCaP) by entering (0, 10, 50 and 100 cells) into the blood of a healthy donor.

For the preparation of RNA samples, blood samples were taken from patients participating in the clinical study and sent to the contracting center of Janssen Pharmaceutical KK, Japan. RNA was extracted from frozen cell lysate samples from clinical trial patients using the Qiagen RNeasy® Plus Micro kit (part number 74034) following the kit manufacturer's instructions, followed by cDNA synthesis using the High Capacity cDNA Reverse Transcription kit (Thermo Fisher Scientific, p/n 4368814) with RNase inhibitor following the manufacturer's instructions. Pre-amplification was performed using the Taqman Pre-amp mastermix (Thermo Fisher Scientific, cat. no. 4391128) reaction mixture following the manufacturer's instructions, followed by PCR analysis using ABI's (Applied Biosystems) Taqman primer/probe pairs. RNA samples were analyzed and normalized using the RPL19 gene (housekeeping gene; NCBI gene no: 6143), positive gene expression was defined as the level of expression above the maximum for controls from healthy patients (59 samples) (i.e. for positive gene expression took on an mRNA expression level greater than the maximum expression level seen in cancer-free mCRPC patients).

Patients.

Ninety-five patients participated and were treated in the two studies (48 chemo-naive and 47 chemo-naive patients in studies JPN-201 and JPN-202, respectively). The biomarker analysis included all patients treated during the study for whom data were available for at least one clinical endpoint and biomarker data suitable for evaluation were available (Table 2). All patients from the JPN-201 and JPN 202 studies were eligible for inclusion in the biomarker analysis dataset. Biomarkers were excluded from the analysis based on the following predefined criteria:

to analyze the level of expression of CTC proteins: if there were less than 3 CTCs suitable for evaluation in the sample, the sample was considered unsuitable for evaluation and excluded from the analysis; and for all mRNAs of CTC biomarkers: if at least one of the results of measuring GAPDH or PSA turned out to be negative, the sample was considered unsuitable for evaluation for all tested biomarkers and excluded from the analysis.

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Table 2. Patient samples available for biomarker analysis

JPN-201 JPN-202 Total Randomized and treated patients, η 48 47 95 Baseline Number of blood samples, η 48 47 95 Counting the number of CTCs 48 47 95 RT-PCR 46 47 93 CL Number of blood samples, η 43 31 74 Counting the number of CTCs 36 25 61 RT-PCR 35 24 59 ^a Baseline and CL Counting the number of CTCs 36 25 61 RT-PCR 34 24 58

59 control samples from healthy patients were also available for analysis of mRNA biomarkers.

Data for patients experiencing serious adverse events leading to discontinuation of treatment were excluded from the analysis. Below are the results of the analysis for those patients who remained on treatment.

To assess the association of dichotomized categorical changes in biomarkers from baseline to CL, the paired McNemar test was used, and to assess continuous changes in biomarkers from baseline to CL, the paired Wilcoxon rank sum test was used. An associative analysis of biomarkers and biomarker changes with clinical response was performed in the pooled JPN-201 and JPN-202 studies and in each study separately. Logistic regression or Fisher's exact test was used to evaluate association with biomarker levels at baseline or in CL with clinical endpoints or association with change in biomarker levels from baseline to CL with PSA response. To correct for interpatient variability in CTC counts, CTC counts were also included in the analysis of the association of gene expression levels with clinical outcomes. The association of biomarker levels with endpoints determined by time to event, overall survival, rPFS, and PSA-PFS was assessed using a Cox proportional hazards model adjusted for the number of CTCs. Survival analyzes were performed for endpoints defined by time to event, adjusted for the number of CTCs. Uncorrected P-values are reported for all analyzes.

The effect of treatment, the level of expression of the biomarker in CL compared with the baseline, is presented in the table below. 3.

Table 3. Biomarker gene expression level at the end of treatment compared to baseline

Biomarker Baseline CL % change from baseline p value negative I Positive ARFL Positive negative 8 20 21 9 2% 1,000 ARV1 Positive negative 4 31 2 21 29% 0.001 ARV7 Positive negative 4 26 eleven 17 22% 0.009 PSMA Positive negative 0 16 25 eleven 21% 0.003 CDH1 Positive negative 3 25 15 9 12% 0.149 IFIH1 Positive negative 3 46 1 2 -2% 1,000

The level of gene expression was dichotomized using the maximum levels found in controls from healthy donors. Baseline levels are given in rows, CL levels are given in columns, p-values are given as unadjusted p-values.

Changes in gene expression levels from baseline to CL/CI were analyzed and the results obtained for the genes of interest are presented in FIG. 1A, fig. 1B and in table. 3. When analyzing

- 9 042117 pooled study data, the proportion of patients with expressed PSMA was higher in CL relative to baseline (69% vs. 48%, P=0.003; Fig. 1A). There were no significant changes from baseline in the proportion of patients expressing AR FL (52% compared to 50%, P = 1.0), while there were more patients with ARV1 expression and ARV7 expression in CL compared with baseline (40 % versus 10% and 48% versus 26%, respectively, P<0.01) (FIG. 1B). ARV567 expression levels in most patients' samples were undetectable either at baseline or in CL (data not shown).

The association of biomarkers at baseline with clinical response and resistance was assessed. In table. 4 shows the main performance indicators (PSA response) at baseline, and in table. 5 shows the PSA response in CL.

Table 4. Association analysis of biomarkers at baseline with PSA response for pooled data from JPN-201 and JPN-202 studies

Biomarker No answer ³ , η (%) Answer ³ , η (%) Total, η (%) Disagreement Rate (95% CI) Meaning R Expression of ARV7 ^b Negative Positive 28 (74) 10 (26) 25 (76) 8 (24) 53 (76) 18 (25) 0.897 (0.2622.987) 1,000 Expression of ARVl ^b Negative Positive 32 (84) 6 (16) 29 (88) 4 (12) 61 (86) 10 (14) 0.739 (0.1393.480) 0.742 Full length AR expression ^b Negative Positive 14 (37) 24 (63) 19 (58) 14 (42) 33 (46) 38 (54) 0.435 (0.1481.235) 0.099 RZMA expression ^b Negative Positive 12 (36) 21 (64) 20 (69) 9 (31) 32 (52) 30 (48) 0.263 (0.0780.833) 0.013

^and the PSA response is defined as a >50% drop in PSA levels by week 12, ^and gene expression levels were dichotomized separately for each gene using the maximum levels observed in controls from healthy donors, including all RNA samples in the analysis, regardless of the number of CTCs.

Frequencies are shown by column as fractions.

The P values are based on Fisher's exact test.

Table 5. Association analysis of biomarkers in CL with PSA response for pooled data from studies JPN-201 and JPN-202

Biomarker No answer ³ , η (%) Answer ³ , η (%) Total, η (%) Disagreement Rate (95% CI) Meaning R Expression of ^ARV7b 11 (35) 20 (65) 18 (69) 8 (31) 29 (51) 28 (49) 0.251 (0.069, 0.842) 0.017

- 10042117

negative Positive Expression of ARVl ^b Negative Positive 15 (48) 16 (52) 19 (73) 7 (27) 34 (60) 23 (40) 0.352 (0.096, 1.193) 0.103 Expression of total ARV ^C Negative Positive 9 (29) 22 (71) 17 (65) 9 (35) 26 (46) 31 (54) 0.223 (0.061, 0.755) 0.008 RZMA expression ^b Negative Positive 4 (14) 25 (86) 12 (52) 11 (48) 16 (31) 36 (69) 0.153 (0.029, 0.646) 0.006 Expression of NPY ^b Negative Positive 12 (41) 17 (59) 18 (78) 5 (22) 30 (58) 22 (42) 0.203 (0.045, 0.772) 0.011 Expression of ACADL ^b Negative Positive 14 (48) 15 (52) 16 (70) 7 (30) 30 (58) 22 (42) 0.416 (0.109, 1.471) 0.162 Expression of UBE2C ^b Negative Positive 26 (90) 3 (10) 22 (96) 14) 48 (92) 4 (8) 0.4 (0.007, 5.402) 0.621

^and no PSA response is defined as a >50% drop in PSA levels by week 12, ^b gene expression levels were dichotomized separately for each gene using the maximum levels observed in controls from healthy donors, including all RNA samples in the analysis, regardless of the number of CTCs, ^c total AR expression is defined as positive expression of any one of the ARV1, ARV7, or ARV567 variants.

Frequencies are shown by column as fractions.

The P values are based on Fisher's exact test.

Regarding associations of gene expression with the primary endpoint, PSMA expression levels at baseline and in CL were associated with lower PSA response rates (at baseline: 30% in PSMA+ versus 63% in PSMA-, P=0.01 CL: 31% in PSMA+ versus 75% in PSMA-, P=0.01). AR FL expression levels at baseline and in CL had no significant effect on PSA response (37% in AR FL+ vs 58% in AR FL-, P=0.10 and 43% in AR FL+ vs 48% in AR FL-, P=0.19, respectively). ARV1 or ARV7 expression levels at baseline had no significant effect on PSA response (40% in ARV1+ versus 48% in ARV1-, P = 0.14; 44% in ARV7+ versus 47% in ARV7-, P =1.0). The expression level of ARV7 in CL was associated with a lower PSA response (29% in ARV7+ versus 62% in ARV7-, P<0.02). Positive expression of NPY in CL was associated with a lower PSA response rate (22% responders versus 59% non-responders, p=0.011).

Of all patients sampled at baseline for biomarker analysis, 44% had >5 CTCs (Table 6). Of patients with CTCs >5 at baseline, 34% to 54% transitioned to CTCs<5 during treatment and 20% retained CTCs<5 in CL/CT. The majority of samples with CTC<5 at baseline (56% at baseline) retained CTC<5 during treatment (89-94%) until progression (41% in CL/CT).

Table 6. Number (Percentage) of Patients in Classes with Different CTC Conversions at Each Time Point Compared to Baseline, Pooled Data from JPN-201 and JPN-202

Category by CSC C1D1 C2DK5 C3DK5 C4DK5 CL or CI < 5 < 5 38/68 (56) 33/35 (94) 36/37 (97) 32/36 (89) 11/27 (41) > 5 30/68 (44) 10/29 (34) 12/29 (41) 14/26 (54) 4/20 (20) Total < 5 38/68 (56) 43/64 (67) 48/66 (73) 46/62 (74) 15/47 (32)

With respect to associations of gene expression with secondary endpoints (Fig. 2A and Fig. 2B), the level of PSMA expression at baseline was associated with shorter rPFS, PFS

- 11 042117 and OS (KH, 2.3, 3.2 and 4.9, respectively, P<0.05) (Fig. 2A). PSMA expression level in CL was associated with a shorter PFS value (KH, 3.1, P=0.04) (Fig. 2A). The level of AR FL expression at baseline was associated with shorter PFS (KH, 2.5, P=0.004) and shorter rPFS (KH, 1.77, P=0.048; Fig. 2B). No evidence of significant association between AR FL expression in CL and rPFS, PFS, or OS values (KH, 1.46, P=0.25; KH, 1.90, P=0.06; and KH, 0.63, P=0.32 respectively) was not detected (Fig. 2B). ARV7 expression level at baseline did not show a significant association with rPFS, PFS, or OS values, while ARV7 expression level in CL showed a significant association only with a shorter PFS value (KH, 2.7, P=0.01; Fig. 2B ). The expression level of CDH1 in CL was associated with a shorter PFS value (KH, 2.32, P=0.04), and IFIH1 was associated with a shorter rPFS value (KH, 4.54, P=0.06) (Table 7 ). Positive expression of UBE2C (proliferation marker) in CL was associated with shorter OS (KH, 3.70, P=0.03), and ACADL (androgen-related fatty acid β-oxidation enzyme) with shorter PFS (KH , 2.48, P=0.02) (Table 7).

Table 7. Association of gene expression levels in CL with secondary clinical endpoints

OS PFS rPFS Biomarker ³ KN (CI 95%) Meaning R KN (CI 95%) Meaning R KN (CI 95%) Meaning P^ CDH1 0.69 (0.23-2.06) 0.50 2.32 (1.03-5.23) 0.04 1.41 (0.68-2.91) 0.36 IF1H1 1.25 (0.16-9.66) 0.83 0.46 (0.11-1.98) 0.30 4.54 (0.94-22.07) 0.06 UBE2C 3.70 (1.17-11.68) 0.03 1.25 (0.40-3.93) 0.70 1.87 (0.60-5.78) 0.28 NPY 0.85 (0.26-2.76) 0.78 1.03 (0.50-2.11) 0.94 0.65 (0.31-1.35) 0.25 ACADL 0.73 (0.26-2.08) 0.56 2.48 (1.17-5.29) 0.02 1.11 (0.57-2.20) 0.75

a The assessed genes were dichotomized as positive or negative for expression based on cut-off values established from analyzes of normal healthy patients, ^b P values based on Fisher's exact test.

Analysis of pooled study data showed no significant changes in AR protein expression in CL relative to baseline (Table 8), and no significant changes (CL relative to baseline) were found in absolute amounts of AR+ and AR- CTCs. An inconsistency was observed between AR FL+ gene expression and AR+ protein expression (Table 9); Sixteen of 18 (89%) AR+ CTC patient samples were classified as AR FL+, while 10 of 12 (83%) AR-CTC patient samples were classified as AR FL+. Positive AR expression at baseline and in CL had no significant effect on PSA response (37% in AR+ versus 36% in AR-, P=1.0 and 36% in AR+ versus 37% in AR-, P=1.0, respectively) (Table 9). AR expression, neither at baseline nor in CL, was significantly associated with secondary clinical endpoints (P>0.05); however, there was a tendency to associate baseline AR expression with improvement in rPFS in patients treated with AA plus prednisone (KN, 0.56, P=0.17; Table 10).

Table 8. Change in the number of CTCs from baseline to CL in AR+ and AR- samples

Initial level (l=31) CL (n=33) Biomarker ³ Median value (range) Median value (range) P ^b value AR+ 3 (0-73) 1 (0-45) 0.88 AR- 7 (0-530) 17 (3-1069) 0.47

^and AR protein expression in CTCs was dichotomized as <10% (negative) or >10% (positive). Unpaired expression data included, ^b P values based on Wilcoxon's paired rank sum test.

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Table 9. Comparison of positive AR protein expression and _________ AR gene expression at baseline_________

Expression AR ^a gene Expression of AR ^b protein Positive negative Positive 16 2 negative 10 2

a _ _ _ _ _ _ _ _ _ _ _ _ _ ___ The assessed genes were dichotomized as positive or negative for expression based on thresholds established from analysis of normal healthy patients, ^b AR protein expression in the CTC was dichotomized as <10% (negative) or >10% (positive). Patients with <5 CTCs were excluded from the analysis.

Table 10 Association of AR protein expression at baseline and in CL with secondary clinical endpoints

OS PFS rPFS Biomarker ³ KN (CI 95%) A value KN (CI 95%) Meaning A KN (CI 95%) P° value AR (IS) 0.70 (0.27- 1.81) 0.46 0.90 (0.42-1.92) 0.79 0.56 (0.24-1.28) 0.17 AR (CL) 0.62 (0.24- 1.59) 0.32 1.22 (0.56-2.65) 0.62 0.92 (0.44-1.91) 0.82

^and AR protein expression in CTCs was dichotomized as <10% or >10%. Patients with >5 CTCs were excluded from the analysis, ^b P values are based on a Cox proportional hazards model, with the number of CTCs being a covariate.

Description.

The overall goal of the biomarker strategy was to identify biomarker profiles that would be associated with sensitivity or resistance to AA plus prednisone therapy for mCRPC cancer in patients in Japan. Since the choice of further therapy in resistant to AA plus prednisone therapy presents significant difficulties, such an analysis can provide biomarker indicators for choosing the optimal therapy for individual patients, based on their molecular profile. This biomarker analysis also has broader implications for clinical practice as it provides a possible approach to conduct a comprehensive study of AR axis centered CTC biomarkers and for the first time offers a way to assess AR and its splicing patterns in CTCs.

Baseline PSMA expression levels and CL levels have been identified as potential biomarkers associated with worse outcomes in patients with mCRPC cancer. Positive expression of PSMA was noted more frequently in non-responders in the pooled population of mCRPC patients and chemotherapy-naive patients. In addition, for chemotherapy-naive patients, an association was observed between PSMA positive expression at baseline and shorter PFS and rPFS values. It is possible to use PSMA expression as a biomarker for decision making in the choice of therapy following AA plus prednisone therapy and in developing therapeutic strategies for patients with mCRPC, especially those who have not previously received chemotherapy.

Although ARV7 expression level in CL was associated with a lower PSA response (29% in ARV7+ versus 62% in ARV7-, P<0.02) and shorter PFS (KN, 2.7, P=0.01) , baseline ARV7 expression did not show an association with clinical outcomes. Positive CTC nuclear staining for AR or expression of wild-type AR or AR splicing variants did not show association with PSA response, indicating the existence of a primary AA resistance mechanism and heterogeneity of AR signaling pathways. These results contradict recent reports on the possibility of an association of ARV7 expression levels in patients with metastatic castration-resistant prostate cancer with resistance to abiraterone (Antonarakis E.S., et al. AR-V7 and resistance to enzalutamide and abiraterone in prostate cancer. N Engl J Med 2014 ;371:1028-1038;Qu F, et al.Association of AR-V7 and prostate specific antigen RNA levels in blood with efficacy of abiraterone acetate and enzalutamide treatment in men with prostate cancer.Clin Cancer Res 2017;23:726-34 Del Re M., et al., The detection of androgen receptor splice variant 7 in plasma-derived exosomal RNA

- 13 042117 strongly predicts resistance to hormonal therapy in metastatic prostate cancer patients. EUR Urol 2017; 71:680-7; and Todenhofer T, et al. AR-V7 transcripts in whole blood RNA of patients with metastatic castration resistant prostate cancer correlate with response to abiraterone acetate. J Urol 2017; 197:135-42). Among abiraterone acetate-treated men, ARV7-positive patients had lower PSA response rates compared with ARV7-negative patients (0% vs. 68%, p = 0.004) and lower PSA-PFS (median, 1.3 months vs. with no progression; p<0.001), clinical or radiographic progression-free survival (median, 2.3 months vs. no; p<0.001), and overall survival (median, 10.6 months vs. no achieved, p=0.006). In addition, high blood levels of ARV7 expression at baseline have been reported to predict poorer OS outcomes in AA-treated patients with mCRPC cancer. In one study evaluating a novel extraction method, extracting RNA from plasma-derived exosomes, OS was significantly lower in participants classified as ARV7+ at baseline compared to ARV7- classified patients with CRPC cancer treated with AA or enzalutamide (8 months after compared with not achieved, P<0.001). However, another study also showed that some groups of patients with CRPC cancer may benefit from AA plus prednisone and/or enzalutamide despite the identification of ARV7 splicing variants in their CTCs (Bernemann C, et al., Expression of AR-V7 in circulating tumor cells does not preclude response to next generation androgen deprivation therapy in patients with castration resistant prostate cancer. Eur Urol 2017; 71: 1-3). When combining the results of the two studies, higher rates of PSMA-positive patients were observed in PSA non-responders compared to PSA-responders.

Inconsistency was observed between mRNA and AR protein expression and association analyzes between these biomarkers and clinical outcomes. In such an assay, mRNA detection is more sensitive than protein detection. Baseline AR FL expression was correlated with worse clinical outcomes and showed a significant association with shorter PFS (KN, 2.5, P=0.004) in patients treated with AA plus prednisone, with baseline AR protein expression showing a trend to improve clinical outcomes, in particular the trend towards an increase in rPFS (HK, 0.56, P=0.17). The inconsistencies in these results may be due to the dependence of AR expression level on the clinical benefit of AA plus prednisone therapy, since the level of AR FL expression in patients in this analysis did not correlate with AR expression. AA suppressed the expression of genes (eg, IFIH1, CDH1) that are downregulated in docetaxel-resistant tumors. These genes were identified in a exploratory microarray assay for docetaxel resistance in two CRPC cancer cell lines (22). The epithelial cell adhesion molecule CDH1 is associated with the epithelial-mesenchymal transition process, and the IFIH1 molecule has been associated with antiviral cellular responses (22).

Conclusions.

Higher frequencies of PSMA at baseline and in CL were associated with worse clinical outcomes (i.e., it is a biomarker of AA resistance in mCRPC patients, especially in chemo-naive mCRPC patients). The initial level of expression of ARv7 in CL showed a weak association with clinical results. The initial level of expression of NPY, UBE2C and ACADL in CL was associated with worse clinical outcomes. AA suppressed the expression of the IFIH1 and CDH1 genes, which are characterized by a decrease in docetaxel-resistant tumors.

Those skilled in the art will appreciate that the preferred embodiments of the present invention are susceptible to many changes and modifications, and that such changes and modifications may be made without departing from the spirit of the invention. Thus, the appended claims are intended to include all such equivalent variations as fall within the true spirit and scope of the present invention.

The descriptions of each patent, patent application, and publication cited or described in this document are incorporated herein by reference in their entirety.

Embodiments

The following list of embodiments is intended to supplement, not replace or invalidate the preceding descriptions.

Embodiment 1. A method for diagnosing and treating metastatic castration-resistant prostate cancer resistant to abiraterone acetate glucocorticoid therapy in a patient, comprising:

for a patient treated with abiraterone acetate and glucocorticoid, diagnosing abiraterone acetate-glucocorticoid therapy-resistant metastatic castration-resistant prostate cancer in a patient with elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample; and treating the diagnosed patient using a therapeutically effective amount of a therapeutic agent other than abiraterone acetate and a glucocorticoid, or using

- 14 042117 abiraterone acetate and glucocorticoid in combination with a therapeutically effective amount of an additional therapeutic agent.

Embodiment 2. A method for diagnosing and treating abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer in a patient, comprising:

for a patient treated with abiraterone acetate and a glucocorticoid, diagnosing abiraterone acetate-glucocorticoid therapy-responsive metastatic castration-resistant prostate cancer in a patient when the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample is not elevated; and treating the diagnosed patient with a therapeutically effective amount of abiraterone acetate and a glucocorticoid.

Embodiment 3. The method of Embodiment 1 or 2, further comprising determining the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in a patient biological sample prior to diagnosis.

Embodiment 4. The method of Embodiment 3 wherein the biological sample contains circulating tumor cells.

Embodiment 5. The method of any one of the preceding embodiments, wherein the level of PSMA, ACADL, NPY, or UBE2C is the mRNA level of PSMA, ACADL, NPY, or UBE2C.

Embodiment 6. The method of any one of the preceding embodiments, further comprising comparing the level of PSMA, ACADL, NPY, UBE2C or any combination thereof with a control prior to diagnosis.

Embodiment 7. The method of Embodiment b wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C or any combination thereof from the patient.

Embodiment 8. The method of Embodiment 6 wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C or any combination thereof from a patient or patient population that does not have metastatic castration resistant prostate cancer.

Embodiment 9. A method for detecting resistance or sensitivity to abiraterone acetate-glucocorticoid therapy in a patient having metastatic castration-resistant prostate cancer, comprising:

determination of the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in a biological sample from a patient treated with abiraterone acetate and glucocorticoid, and an increased level of PSMA, ACADL, NPY, UBE2C, or any combination of them indicates acquired resistance to abiraterone acetate-glucocorticoid therapy, and however, the absence of elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination of them indicates sensitivity to abiraterone acetate-glucocorticoid therapy.

Embodiment 10. The method of Embodiment 9 wherein the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C.

Embodiment 11. The method of Embodiment 9 or 10 wherein the patient has or does not have an elevated level of PSMA, ACADL, NPY, UBE2C or any combination thereof compared to a control.

Embodiment 12. The method of Embodiment 11 wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C or any combination thereof from the patient.

Embodiment 13. The method of Embodiment 11 wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C or any combination thereof in a patient or patient population that does not have metastatic castration resistant prostate cancer.

Embodiment 14. A method of treating metastatic castration-resistant prostate cancer, comprising: administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to a patient having mCRPC cancer, wherein the patient does not have elevated levels of ACADL, NPY, UBE2C, or any combination thereof.

Embodiment 15. The method of Embodiment 14 wherein the patient has previously been treated with abiraterone acetate and a glucocorticoid.

Embodiment 16. The method of Embodiment 15 wherein the patient does not have an elevated PSMA level.

Embodiment 17. The method of any one of Embodiments 14-16 wherein the abiraterone acetate and the glucocorticoid are administered simultaneously or sequentially.

Embodiment 18. The method of any one of Embodiments 14-17, wherein the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C.

Embodiment 19. The method of any one of Embodiments 14-18, wherein the patient does not have an elevated level of PSMA, ACADL, NPY, UBE2C, or any combination thereof compared to a control.

Embodiment 20. The method of Embodiment 19 wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C or any combination thereof from the patient.

Embodiment 21. The method of Embodiment 20 wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C or any combination thereof in a patient or patient population that does not have metastatic castration resistant prostate cancer.

Embodiment 22. A method of treating metastatic castration-resistant prostate cancer, comprising: administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to a patient who has previously received abiraterone acetate and glucocorticoid therapy, wherein the patient does not have elevated PSMA, ACADL, NPY, UBE2C, or any combination of them.

Embodiment 23. The method of Embodiment 22 wherein the abiraterone acetate and the glucocorticoid are administered simultaneously or sequentially.

Embodiment 24. The method of Embodiment 22 or 23 wherein the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C.

Embodiment 25. The method of any one of Embodiments 22-24, wherein the patient does not have elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof compared to a control.

Embodiment 26. The method of Embodiment 25 wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C or any combination thereof from the patient.

Embodiment 27. The method of Embodiment 25 wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C or any combination thereof in a patient or patient population that does not have metastatic castration resistant prostate cancer.

Embodiment 28. A method of treating metastatic castration-resistant prostate cancer, comprising: for a patient not having elevated levels of ACADL, NPY, UBE2C, or any combination thereof, administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to treat metastatic castration-resistant cancer prostate.

Embodiment 29. The method of Embodiment 28 wherein the patient has previously been treated with abiraterone acetate and a glucocorticoid.

Embodiment 30. The method of Embodiment 29 wherein the patient does not have an elevated PSMA level.

Embodiment 31. The method of any one of Embodiments 28-30 wherein the abiraterone acetate and the glucocorticoid are administered simultaneously or sequentially.

Embodiment 32. The method of any one of Embodiments 28-31, wherein the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C.

Embodiment 33. The method of any one of Embodiments 28-32, wherein the patient does not have elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof compared to a control.

Embodiment 34. The method of Embodiment 33 wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C or any combination thereof from the patient.

Embodiment 35. The method of Embodiment 34 wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C or any combination thereof in a patient or patient population that does not have metastatic castration resistant prostate cancer.

Embodiment 36. A method of treating metastatic castration-resistant prostate cancer, comprising: for a patient not having elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof, administering a therapeutically effective amount of abiraterone acetate and a therapeutically effective amount of a glucocorticoid to treat metastatic castration- resistant prostate cancer.

Embodiment 37. The method of Embodiment 36 wherein the abiraterone acetate and the glucocorticoid are administered simultaneously or sequentially.

Embodiment 38. The method of Embodiment 36 or 37 wherein the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C.

Embodiment 39. The method of any one of Embodiments 36-38, wherein the patient does not have elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof compared to a control.

Embodiment 40. The method of Embodiment 39 wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C or any combination thereof from the patient.

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Embodiment 41. The method of Embodiment 40 wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C or any combination thereof in a patient or patient population that does not have metastatic castration resistant prostate cancer.

Embodiment 42 Use of PSMA, ACADL, NPY, UBE2C or any combination thereof in predicting sensitivity or acquired resistance to abiraterone acetate glucocorticoid therapy in a patient with metastatic castration resistant prostate cancer.

Embodiment 43 PSMA, ACADL, NPY, UBE2C or any combination thereof for use in predicting sensitivity or acquired resistance to abiraterone acetate glucocorticoid therapy in a patient with metastatic castration resistant prostate cancer.

Embodiment 44 The use of embodiment 42 or 43 wherein PSMA, ACADL, NPY, UBE2C or any combination thereof is mRNA obtained from a biological sample from a patient and the patient has previously received abiraterone acetate and glucocorticoid therapy.

Embodiment 45. The method of any one of Embodiments 1-41, or the use of any one of Embodiments 42-44, wherein the glucocorticoid is prednisone, prednisolone, hydrocortisone, dexamethasone, cortisone, or methylprednisolone.

Claims (21)

1. A method of treating abiraterone acetate-prednisone-responsive metastatic castration-resistant prostate cancer in a patient, comprising: for a patient treated with abiraterone acetate 1000 mg/day and prednisone 10 mg/day, a diagnosis of abiraterone acetate-prednisone therapy-responsive metastatic castration-resistant prostate cancer in a patient when PSMA, ACADL, NPY, UBE2C, or any of these combinations in the biological sample of the patient is not elevated; and treating the diagnosed patient with abiraterone acetate and prednisone. 2. The method of claim 1, further comprising determining the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in the patient's biological sample prior to diagnosis. 3. The method of claim 2, wherein the biological sample contains circulating tumor cells. 4. The method according to any one of the preceding claims, wherein the level of PSMA, ACADL, NPY or UBE2C is the mRNA level of PSMA, ACADL, NPY or UBE2C. 5. The method according to any one of the preceding claims, further comprising comparing the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof with a control prior to diagnosis. 6. The method of claim 5, wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from the patient. 7. The method of claim 5, wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, from a patient or patient population that does not have metastatic castration-resistant prostate cancer. 8. A method of treating metastatic castration-resistant prostate cancer in a patient, comprising administering abiraterone acetate and prednisone to a patient to treat metastatic castration-resistant prostate cancer, wherein the patient does not have elevated levels of ACADL, NPY, UBE2C, or any combination thereof. 9. The method of claim 8 wherein the patient has previously been treated with abiraterone acetate and prednisone. 10. The method of claim 9, wherein the patient does not have an elevated PSMA level. 11. The method according to any one of claims 8 to 10, wherein the abiraterone acetate and prednisone are administered simultaneously or sequentially. 12. The method according to any one of claims 8-11, wherein the level of PSMA, ACADL, NPY, UBE2C or any combination thereof is the mRNA level of PSMA, ACADL, NPY or UBE2C. 13. The method of any one of claims 8-12, wherein the patient does not have elevated levels of PSMA, ACADL, NPY, UBE2C, or any combination thereof compared to a control. 14. The method of claim 13, wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from the patient. 15. The method of claim 13, wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C, or any combination thereof, in a patient or patient population that does not have metastatic castration-resistant prostate cancer. 16. A method of treating metastatic castration-resistant prostate cancer in a patient, comprising administering abiraterone acetate and prednisone to a patient for the treatment of metastatic castration-resistant prostate cancer, wherein the patient has previously been treated with abiraterone acetate at a dose of 1000 mg/day and prednisone at a dose of 5 mg/day , and at the same time, the patient has - 17 042117 no elevated PSMA, ACADL, NPY, UBE2C or any combination thereof. 17. The method of claim 16 wherein the abiraterone acetate and prednisone are administered simultaneously or sequentially. 18. The method of claim 16 or 17, wherein the level of PSMA, ACADL, NPY, UBE2C, or any combination thereof is the mRNA level of PSMA, ACADL, NPY, or UBE2C. 19. The method of any one of claims 16-18, wherein the patient does not have an elevated level of PSMA, ACADL, NPY, UBE2C, or any combination thereof compared to a control. 20. The method of claim 19, wherein the control contains a prior level of PSMA, ACADL, NPY, UBE2C, or any combination thereof from the patient. 21. The method of claim 19, wherein the control comprises a level of PSMA, ACADL, NPY, UBE2C, or any combination thereof in a patient or patient population that does not have metastatic castration-resistant prostate cancer.