KDM3a inhibitors for treating colon cancer and prostate cancer

Abstract

Provided herein are compounds for inhibiting cancer cell proliferation and methods for treating cancer and improving the therapeutic effect of an androgen receptor signaling inhibitor utilizing such compounds.

Description

KDM3A INHIBITORS FOR TREATING COLON CANCER AND PROSTATE CANCERCROSS-REFERENCE TO RELATED APPLICATION

[0001] The present application claims priority to U.S. Provisional Patent Application Serial No. 63 / 637,642, filed on April 23, 2024, the entire disclosure of which is incorporated herein by reference.TECHNICAL FIELD

[0002] The present disclosure relates to compounds useful for the treatment of cancer. In particular, certain embodiments of the presently disclosed subject matter relate to compounds that inhibit histone demethylase 3A (KDM3A) and the use thereof to inhibit cancer cell proliferation and / or improve the therapeutic effect of an androgen receptor signaling inhibitor.BACKGROUND

[0003] Prostate cancer is the second leading cause of cancer-related mortality among men in the United States. Androgen receptor (AR) plays a crucial role in prostate cancer development and progression. Prostate cancer patients commonly receive androgen deprivation therapy (ADT) through surgery (e.g., orchiectomy, medical castration) or drug treatment (e.g., AR inhibitors) for treatment. While patients have an initial response to ADT, they often subsequently develop castration-resistant prostate cancer (CRPC). Enzalutamide, an AR inhibitor, has been approved by the U.S. Food and Drug Administration (FDA) to treat both metastasis and non -metastasis CRPC. However, a high percentage of patients have cancer which is resistant to enzalutamide or which subsequently becomes resistant to enzalutamide following a period of enzalutamidetreatment. Mechanisms causing enzalutamide resistance include AR overexpression or amplification, and mutation or expression of AR variants, particularly androgen receptor splice variant (AR-V7). Activation of other signaling pathways, such as Wnt / b-catenin and PI3K pathways also contribute to enzalutamide resistance.

[0004] Colorectal cancer (CRC) is one of the most common cancers in the U.S. and contributes to thousands of deaths each year. A majority of CRC cases involve mutations in the Wnt signaling pathway. Accordingly, Wnt signaling in CRC progression makes it an important, potential target for the development of new anti cancer agents.SUMMARY

[0005] The presently disclosed subject matter meets some or all of the above-identified limitations, as will become evident to those of ordinary skill in the art after a study of information provided in this document.

[0006] This summary describes several embodiments and implementations of the presently disclosed subject matter, and in many cases lists variations and permutations of these embodiments and implementations. This summary is merely exemplary of the numerous and varied embodiments and implementations. Mention of one or more representative features of a given embodiment or implementation is likewise exemplary. Such an embodiment or implementation can typically exist with or without the feature(s) mentioned; likewise, those features can be applied to other embodiments or implementations of the presently disclosed subject matter, whether listed in this summary or not. To avoid excessive repetition, this summary does not list or suggest all possible combinations of such features.

[0007] The presently disclosed subject matter includes compounds for inhibiting cancer cell proliferation. In some embodiments, a compound for inhibiting cancer cell proliferation comprises the structure according to Formula I:or analogs thereof; where R1is branched or unbranched alkyl, amine, or heterocycle. In some

[0008] Also provided herein are methods for inhibiting cancer cell proliferation in a subject.In some implementations, a method for inhibiting cancer cell proliferation comprises administering to the subject a compound selected from the group consisting of:

[0009] In some implementations of the method for inhibiting cancer cell proliferation, the subject to which the compound is administered has a cancer selected from leukemia, lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, and combinations thereof.

[0010] In some implementations of the method for inhibiting cancer cell proliferation, the subject has prostate cancer. In some implementations of the method, the subject has castrationresistant prostate cancer. In some implementations of the method, the subject has androgen receptor axis-targeted therapy (ARAT)-resistant cancer. In some implementations of the method, androgen receptor splice variant 7 (AR-V7) is expressed in a cancer cell present in the subject. In some implementations of the method, the method includes detecting AR-V7 expression in the cancer cell present in the subject. In some implementations of the method, the method includes administering an androgen receptor signaling inhibitor to the subject. In some implementations of the method, the androgen receptor signaling inhibitor is an androgen synthesis inhibitor. Insome implementations of the method, the androgen receptor signaling inhibitor is an androgen receptor antagonist. In some implementations of the method, the androgen receptor signaling inhibitor is enzalutamide. In some implementations of the method, the compound is

[0011] In some implementations of the method for inhibiting cancer cell proliferation, the subject has colon cancer. In some implementations, the compound is

[0012] In some implementations of the method for inhibiting cancer cell proliferation, the subject is a mammal.

[0013] Further provided herein are methods for improving the therapeutic effect of an androgen receptor signaling inhibitor. In some implementations, a method for improving the therapeutic effect of an androgen receptor signaling inhibitor comprises administering to asubject with cancer a compound selected from the group consisting of

[0014] In some implementations of the method for improving the therapeutic effect of an androgen receptor signaling inhibitor, the subject has prostate cancer. In some implementations of the method, the subject has castration-resistant prostate cancer. In some implementations of the method, the subject has ARAT-resistant cancer. In some implementations of the method, AR- V7 is expressed in a cancer cell present in the subject. In some implementations of the method, the androgen receptor signaling inhibitor is an androgen synthesis inhibitor. In some implementations of the method, the androgen receptor signaling inhibitor is an androgen receptor antagonist. In some implementations of the method, the androgen receptor signaling inhibitor is enzalutamide.

[0015] In some implementations of the presently disclosed methods, the compound may be administered as part of a pharmaceutical composition, which includes the compound and a pharmaceutically acceptable carrier. Accordingly, the present disclosure also provides a pharmaceutical composition comprising at least one of the of the compounds provided in the present disclosure, together with a pharmaceutically acceptable carrier.

[0016] Further features and advantages of the presently disclosed subject matter will become evident to those of ordinary skill in the art after a study of the description, figures, and nonlimiting examples in this document.BRIEF DESCRIPTION OF DRAWINGS

[0017] The presently disclosed subject matter will be better understood, and features, aspects, and advantages will become apparent when consideration is given to the following detailed description. Such detailed description makes reference to the following drawings, wherein:

[0018] FIG. 1 is a graph showing the inhibitory effects of VMS-7-118 (identified in FIG. 1 as V-7-118 and also referred to herein as CBA-2) and ZBC-1-83 (identified in FIG. 1 as Z-l-183 and also referred to herein as CBA-3) on lysine demethylase 3A (KDM3A) enzymatic activity at 1 pM and 10 pM concentrations.

[0019] FIG. 2A is an image showing comparative tumor growth of control (top) and VMS-7- 118-treated (10 mg / kg, intraperitoneal (IP)) (bottom) LS174T colorectal cancer (CRC) xenografted severe combined immunodeficient (SCID) mice.

[0020] FIG. 2B is a graph showing tumor volume in control and VMS-7-118-treated (10 mg / kg, IP) LS174T colon cancer xenografted SCID mice over a 12-day period.

[0021] FIG. 3 is a graph comparing the survival rate of control and CBA-3 -treated (2 mg / kg twice per week, oral preparation (OP)) LS174T CRC xenografted SCID mice. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

[0022] FIG. 4A is an image showing comparative tumor growth of control (top) , enzalutamide (ENZ)-treated (20 mg / kg per day, oral administration) (middle), and VMS-7-118- treated (10 mg / kg twice per week, IP) (bottom) 22RV1 prostate cancer xenografted SCID mice.* p < 0.05, ** p < 0.01.

[0023] FIG. 4B is a graph showing tumor volume of control, ENZ-treated, and VMS-7-118- treated 22RV1 prostate cancer xenografted SCID mice over a 15 -day period.

[0024] FIG. 5 is a reaction scheme showing an exemplary synthesis of compound ZBC-1-82.

[0025] FIG. 6 is a reaction scheme showing an exemplary synthesis of compound ZBC-1-83 (CBA-3).

[0026] FIG. 7 is a reaction scheme showing an exemplary synthesis of compound VMS-7- 118 (CBA-2).

[0027] FIG. 8 is a schematic representation of enzalutamide interacting with the ligandbinding domain (LBD) of full-length androgen receptor (AR), and of androgen receptor splice variant 7 that lacks the LBD but maintains the DNA-binding domain (DBD).

[0028] FIG. 9 is an image showing that both CBA-1 and CBA-2 increase histone H3 lysine 9 dimethylation (H3K9Me2) levels in 2RV1 cells, inhibit expression of AR-target gene c-Myc, full-length AR, and AR-V7.

[0029] FIG. 10 is a graph showing relative cell proliferation of dimethyl sulfoxide (DMSO), ENZ-treated (50 pM), CBA-l-treated (0.5 pM), and ENZ and CBA-l-treated (50 pM of ENZ and 0.5 pM of CBA-1) 22RV1 cells.

[0030] FIG. 11 is a graph showing relative cell proliferation of DMSO, ENZ-treated (20 pM), CBA-2-treated (0.1 pM), and ENZ and CBA-2 -treated (20 pM of ENZ and 0.1 pM of CBA-2) 22RV1 cells.

[0031] FIG. 12 is a graph showing relative cell proliferation of DMSO and C ABA-3 -treated (0.1 pM) PC3 cells.

[0032] FIG. 13 is a graph showing relative cell proliferation of DMSO and C ABA-3 -treated(0.1 pM) PC3 cells.DESCRIPTION OF EXEMPLARY EMBODIMENTS

[0009] The details of one or more embodiments of the presently disclosed subject matter are set forth in this document. Modifications to embodiments described in this document, and other embodiments, will be evident to those of ordinary skill in the art after a study of the information provided in this document. The information provided in this document, and particularly the specific details of the described exemplary embodiments, is provided primarily for clearness of understanding and no unnecessary limitations are to be understood therefrom. In case of conflict, the specification of this document, including definitions, will control.

[0010] While the terms used herein are believed to be well understood by those of ordinary skill in the art, certain definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[0011] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the invention(s) belong.

[0012] All patents, patent applications, published applications and publications, GenBank sequences, databases, websites and other published materials referred to throughout the entire disclosure herein, unless noted otherwise, are incorporated by reference in their entirety.

[0013] Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

[0014] Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the presently disclosed subject matter, representative methods, devices, and materials are described herein.

[0015] Following long-standing patent law convention, the terms “a”, “an”, and “the” refer to “one or more” when used in this application, including the claims, unless the context clearly dictates otherwise. Thus, for example, reference to “a polypeptide” includes one or more of such polypeptides, and so forth.

[0016] Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about”. Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and claims are approximations that can vary depending upon the desired properties sought to be obtained by the presently disclosed subject matter.

[0017] As used herein, the term “about,” when referring to a value or to an amount of mass, weight, time, volume, concentration or percentage is meant to encompass variations of in some embodiments ±20%, in some embodiments ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5%, and in some embodiments ±0.1% from the specified amount, as such variations are appropriate to perform the disclosed method.

[0018] As used herein, ranges can be expressed as from “about” one particular value, and / or to “about” another particular value. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is alsodisclosed. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0019] As used herein, the term “treatment” is inclusive of prophylactic treatment and therapeutic treatment. As would be recognized by one of ordinary skill in the art, treatment that is administered prior to clinical manifestation of a condition is prophylactic (i.e., it protects the subject against or reduces the risk of the subject developing the condition). If the treatment is administered after manifestation of the condition, the treatment is therapeutic (i.e., it is intended to diminish, ameliorate, control, or maintain the existing condition and / or side effects associated with the condition). The terms relate to medical management of a subject with the intent to substantially cure, ameliorate, stabilize, or substantially prevent a condition of interest (e.g., disease, pathological condition, or disorder), including but not limited to prophylactic treatment to preclude, avert, obviate, forestall, stop, or hinder something from happening, or reduce the severity of something happening, especially by advance action. As such, the terms treatment or treating include, but are not limited to: inhibiting the progression of a condition of interest; arresting or preventing the development of a condition of interest; reducing the severity of a condition of interest; ameliorating or relieving symptoms associated with a condition of interest; causing a regression of the condition of interest or one or more of the symptoms associated with the condition of interest; and preventing a condition of interest or the development of a condition of interest. The terms include active treatment, that is, treatment directed specifically toward the improvement of a condition of interest, and also includes causal treatment, that is, treatment directed toward removal of the cause of the condition of interest.

[0020] As used herein, the term “effective amount” refers to an amount that is sufficient to achieve the desired prophylactic or therapeutic result or to have an effect on undesiredsymptoms. The specific effective amount for any particular subject may depend upon a variety of factors including the particular risk or condition being treated and the severity of the condition; the specific composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration; the route of administration; the rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of a compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. If desired, the effective daily dose can be divided into multiple doses for purposes of administration. Consequently, single dose compositions can contain such amounts or submultiples thereof to make up the daily dose. The dosage can be adjusted by the individual medical practitioner in the event of any contraindications. Dosage can vary, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of compounds and products. It is appreciated that, in the various methods disclosed herein, the compounds disclosed herein can be administered to the subject in an effective amount.

[0021] As will be recognized by one of ordinary skill in the art, the terms “suppression,” “suppressing,” “suppressor,” “inhibition,” “inhibiting” or “inhibitor” do not refer to a complete elimination of a value in all cases. Rather, the skilled artisan will understand that the term “suppressing” or “inhibiting” refers to a reduction or decrease in a measured value, qualitatively or quantitatively. Such reduction or decrease can be determined relative to a control or a prior status of a subject. In some embodiments, the reduction or decrease relative to a control or the prior status of a subject can be about a 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,97, 98, 99, or 100% decrease.

[0022] As used herein, the term “administering” refers to any method of providing a pharmaceutical preparation to a subject. Such methods are well known to those skilled in the art and include, but are not limited to, oral administration, transdermal administration, intraperitoneal administration, and parenteral administration, including injectable such as intravenous administration, intra-arterial administration, intramuscular administration, and subcutaneous administration. Administration can be continuous or intermittent. In various aspects, a preparation can be administered therapeutically; that is, administered to treat an existing disease or condition. In further various aspects, a preparation can be administered prophylactically; that is, administered for prevention of a disease or condition. In some embodiments, oral administration is used. In some embodiments, intravenous (IV) administration is used.

[0023] The compounds disclosed herein can be administered to a subject using a pharmaceutically acceptable carrier. As used herein, the term “pharmaceutically acceptable carrier” refers to sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, bythe use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid and the like. It can also be desirable to include isotonic agents such as sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents, such as aluminum monostearate and gelatin, which delay absorption. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide, poly(orthoesters) and poly(anhydrides). Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues. The injectable formulations can be sterilized, for example, by filtration through a bacterial -retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable media just prior to use. Suitable inert carriers can include sugars such as lactose.

[0024] As used herein, the term “subject” when referring to a subject having a particular condition or risk thereof, or when referring to a subject in need of treatment for a particular condition or risk thereof, refers to a target of administration, which optionally displays symptoms and / or has risk factors related to the particular condition or risk thereof, as would be considered relevant to one skilled in the medical art of making a diagnosis of the particular condition or risk thereof. As used herein, the term “subject” can refer to a vertebrate, such as a mammal. The termincludes human and veterinary subjects. Thus, the subject can be, by way of non-limiting example, a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, fish, cow, cat, guinea pig, or rodent. The term does not denote a particular age or sex.

[0025] As used herein, an “azacycle” is a nitrogen-containing heterocycle that includes one or more nitrogen atoms. In various embodiments, an azacycle can be saturated, partially unsaturated, or wholly unsaturated.

[0026] As used herein, the term “androgen receptor signaling inhibitor” is inclusive of both (i) androgen synthesis inhibitors that reduce the production of androgens and (ii) androgen receptor antagonists that block the androgen receptor from binding with androgens. Accordingly, in various implementations of the methods disclosed herein involving or relating to the use of an androgen receptor signaling inhibitor, unless indicated otherwise, the androgen receptor signaling inhibitor can be an androgen synthesis inhibitor, such as abiraterone, or an androgen receptor antagonist (which can also be characterized as an androgen receptor inhibitor), such as enzalutamide, apalutamide, or darolutamide.

[0027] The presently disclosed subject matter is based, in part, on the discovery that certain compounds effective for inhibiting lysine demethylase 3A (KDM3A) are also effective with respect to inhibiting cancer cell proliferation in various types of cancers.

[0028] Accordingly, the present disclosure includes compounds for inhibiting cancer cell proliferation and methods for treating cancer which make use of such compounds. In some embodiments, a compound for inhibiting cancer cell proliferation has a structure according toFormula I:where R1is branched or unbranched alkyl, amine, or heterocycle. In some embodiments, the compound according to Formula I includes the structure where R1is an azacycle. In some embodiment, the compound according to Formula I includes the structure where R1is a saturated heterocycle. In some embodiments, the compound according to Formula I includes the structure where R1is branched alkyl. In some embodiments, the compound according to Formula I includes the structure where R1is a tertiary amine. In some embodiments, the compound for inhibiting cancer cell proliferation isome embodiments, the compound for inhibiting cancer cell proliferation i.

[0029] Each respective compound disclosed herein can be administered as part of a pharmaceutical composition that includes the compound and a pharmaceutically acceptable carrier. Pharmaceutical compositions which include one or more compounds disclosed herein in combination with a pharmaceutically acceptable carrier are thus also provided herein. In some embodiments, a pharmaceutical composition comprises: a compound selected from CBA-1,VMS-7-102, CBA-2, ZBC-1-82, and CBA-3; and a pharmaceutically acceptable carrier. Various pharmaceutical composition embodiments in which a different one of CBA-1, VMS-7-102,CBA-2, ZBC-1-82, and CBA-3 is utilized with a pharmaceutically acceptable carrier are contemplated herein.

[0030] Lysine demethylase 3A (KDM3A) is a Jumonji domain-containing demethylase that regulates the demethylation of histone H3’s lysine 9 (H3K9Me2). It has been found that inhibiting KDM3A increases H3 lysine 9 dimethylation (H3K9Me2). Without wishing to be bound by theory, it is believed that Wnt signaling is inhibited by increasing H3K9 methylation on Wnt target gene promotors. Consequently, KDM3A inhibiting compounds have also been found to inhibit cancer cell proliferation. The present inventors have found that both CBA-2 and CBA-3 significantly inhibit KDM3A enzymatic activity, and, in turn, inhibit cancer cell proliferation in various different cancers. Specifically, the present inventors have found that CBA-2 inhibits cell proliferation in colorectal cancer (CRC) and prostate cancer cells, while CBA-3 has surprisingly been discovered to inhibit CRC, prostate, leukemia, lung, central nervous system (CNS), melanoma, ovarian, renal, and breast cancer cells. CBA-1, VMS-7-102, and ZBC-1-182 have also each been found by the present inventors to inhibit cancer cell proliferation in CRC cells, with CBA-1 and VMS-7-102 also being found to inhibit prostate cancer cells. Surprisingly, CBA-2 and VMS-7-102 have each been found to provide greater inhibition of CRC cells and / or prostate cancer cells than CBA-1 at the same dosages, despite CBA-1 having been found to exhibit significantly higher Wnt signaling inhibition than either CBA-2 or VMS-7-102. CBA-3 and ZBC-1-82 have likewise been surprisingly found to provide greater inhibition of CRC and / or prostate cancer cells than CBA-1 at the same dosage.

[0031] Accordingly, in some embodiments, a method for treating cancer in accordance with the present disclosure includes administering a compound selected from the compound embodiments disclosed above to a subject in need thereof to thereby inhibit cancer cell proliferation in the subject. Such method can also be characterized as a method for inhibiting cancer cell proliferation in a subject. In some implementations, the compound administered to

[0032] In some implementations, the inhibition of cancer cell proliferation is characterized by a cessation of tumor growth or a reduction in the rate of tumor growth within a subject. In some implementations, a cessation of tumor growth or a reduction in the rate of tumor growth is assessed by detecting the volume of a tumor within a subject at a first time or first period of time prior to administration of the compound, subsequently detecting the volume of the tumor at a second time or second period of time subsequent to administration of the compound or a series of administrations of the compound, and comparing the detected tumor volumes. Accordingly, in some implementations, a method for inhibiting cancer cell proliferation includes detecting a size of a tumor present within the subject at a first time or first period of time prior to administrationof the compound, detecting the size of the tumor at a second time or second period of time subsequent to administration of the compound or a series of administrations of the compound, and comparing the detected tumor volumes. In some implementations, a reduction in the rate of tumor growth is assessed by comparing the volume of a tumor within the subject following administration of the compound or a series of administrations of the compound with the volume of a tumor within an untreated control subject having the same type of cancer and in the same or similar stage of cancer progression. Accordingly, in some implementations, a method for inhibiting cancer cell proliferation includes detecting the volume of a tumor present within the subject following administration of the compound or a series of administrations of the compound, detecting the volume of a tumor present within and untreated control subject, and comparing the detected tumor volumes. Tumor volume can be assessed utilizing known imaging techniques, including, by way of non-limiting example, CT scan, MRI, ultrasound, mammography, PET scan, or combinations thereof.

[0033] In some implementations, the subject to which the compound is administered has leukemia, lung cancer, colon cancer, CNS, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, or combinations thereof.

[0034] In some implementations, the subject to which the compound is administered has prostate cancer. As will become further evident by the discussion that follows, certain compounds disclosed herein have been surprisingly discovered to inhibit castration-resistant prostate cancer (CRPC). CRPC cells in which certain compounds disclosed herein have been found to inhibit cancer cell proliferation include those which express full-length androgen receptor (AR) as well as those that express androgen receptor splice variant 7 (AR-V7) where the ligand binding domain (LBD) is removed. The absence of the LBD enables the AR to functionwithout its normal activation signal and renders the cancer cells resistant to androgen receptoraxis targeted therapy (ARAT), such as that employing the use of androgen receptor signaling inhibitors like enzalutamide, apalutamide, and darolutamide. Accordingly, in some implementations, the subject to which the compound is administered has CRPC. In some implementations, the subject has ARAT -resistant cancer. In some implementations, AR-V7 is expressed in a cancer cell present in the subject.

[0035] Prostate cancer can be detected and diagnosed in a subject employing known diagnostic techniques, including, by way of non-limiting example: prostate-specific antigen testing; digital rectal exam; prostate biopsy; magnetic resonance imaging (MRI); transrectal ultrasound; computed tomography (CT) scan; bone scan; positron emission tomography (PET) scan; Gleason score; biomarker testing; genetic testing; detection of symptoms associated with prostate cancer; or combinations thereof. CRPC can be detected and diagnosed utilizing one or more of the foregoing techniques and / or the detection of continued progression of prostate cancer in a subject despite testosterone suppression (e g., as a result of castration). AR-V7 expression can be detected utilizing known biomarker detection techniques, such as a liquid biopsy where circulating tumor cells are isolated from the subject’s blood and analyzed for AR- V7 mRNA or protein presence utilizing reverse transcriptase polymerase chain reaction (RT- PCR) or immunocytochemistry. In some cases, ARAT-resistant cancer can be detected and diagnosed by the detection of the expression of AR-V7 and / or the detection of prostate cancer progression subsequent to treatment employing ARAT. In some implementations, the method for inhibiting cancer cell proliferation in a subject further includes detecting and / or diagnosing the subject as having prostate cancer, CRPC, and / or ARAT-resistant cancer. In someimplementations, the method for inhibiting cancer cell proliferation further includes detecting the presence of a cancer cell expressing AR-V7 in the subject.

[0036] The presently disclosed subject matter is further based, in part, on the discovery that certain compounds disclosed herein are effective in inhibiting cancer cell proliferation in androgen-independent cancers (i.e., cancers where cancer cells can grow and progress even in the absence of androgens). Accordingly, in some implementations of inhibiting cancer cell proliferation, the subject to which the compound is administered has androgen-independent cancer.

[0037] The presently disclosed subject is further based, in part, on the discovery that compounds disclosed herein can be administered to improve the therapeutic effect of an androgen receptor signaling inhibitor with respect to cancer cell proliferation in ARAT-resistant cancer. In this regard, it has been found that the administration of CBA-1 in combination with an androgen receptor signaling inhibitor, such as enzalutamide (the structure of which is shown in FIG.8), provides significantly greater inhibition of CRPC cell proliferation than either CBA-1 or the androgen receptor signaling inhibitor alone. Similarly, it has been found that the administration of CBA-2 in combination with an androgen receptor signaling inhibitor, such as enzalutamide, provides significantly greater inhibition of CRPC cell proliferation than either CBA-2 or the androgen receptor signaling inhibitor alone. Due to their structural similarity to CBA-2 and their demonstrated or anticipated ability to inhibit KDM3A, VMS-7-102, ZBC-1-82, and CBA-3 are each believed likely to also improve the therapeutic efficacy of an androgen receptor signaling inhibitor, such as enzalutamide.

[0038] Accordingly, in some implementations, the method for inhibiting cancer cell proliferation includes administering both a compound selected from the compound embodimentsdescribed above and an androgen receptor signaling inhibitor to the subject. In some implementations, treatment of a subject involves the administration of a combination drug therapy. In some implementations, the compound administered in combination with the androgen receptor signaling inhibitor is selected from CBA-1, VMS-7-102, CBA-2, ZBC-1-82, and CBA- 3. In some implementations, the androgen receptor signaling inhibitor is selected from abiraterone, enzalutamide, apalutamide, and darolutamide. In some implementations, the androgen receptor signaling inhibitor is enzalutamide and the compound is CBA-1 or CBA-2. In some implementations, the compound is administered at least two times per week and the androgen receptor signaling inhibitor is administered daily. However, the dosage schedule of the compound and the androgen receptor signaling inhibitor can, of course, be modified, for example, to accommodate different subjects, subject response to a particular dosage schedule, different types of cancer, and / or different stages of cancer.

[0039] In some implementations, the subject to which the compound is administered has colorectal cancer. In some implementations, the subject to which the compound is administered has colon cancer. Colon cancer can be detected and diagnosed in a subject employing known diagnostic techniques, including, by way of non-limiting example: colonoscopy; fecal immunochemical test; fecal occult blood test; flexible sigmoidoscopy; CT coIonography; biopsy analysis; detection of symptoms associated with colon cancer; CT scan; MRI; PET scan; biomarker testing; genetic testing; or combinations thereof. In some implementations, the method for inhibiting cancer cell proliferation further includes detecting and / or diagnosing the subject as having colon cancer.

[0040] In some implementations, the compound or combination of the compound and the androgen receptor signaling inhibitor is administered for a predetermined treatment period. Insome implementations, the predetermined treatment period corresponds to a period starting at the time the subject is identified as needing treatment and first administered the compound or combination of the compound and androgen receptor signaling inhibitor and ending at the time when cancer cell proliferation has ceased or a physician determines treatment utilizing the compound or the combination of the compound and an androgen receptor signaling inhibitor is not required for a desired health outcome.

[0041] As reflected in the discussion above, further provided herein are methods for improving the therapeutic effect of an androgen receptor signaling inhibitor. In an exemplary implementation, a method for improving the therapeutic effect of an androgen receptor signaling inhibitor includes administering to a subject with cancer a compound selected from CBA-1, VMS-7-102, CBA-2, ZBC-1-82, and CBA-3 to the subject. In some implementations, the selected compound is administered at least twice per week and the administration of the androgen receptor signaling inhibitor is administered daily. Again, however, the dosage schedule of the compound and the androgen receptor signaling inhibitor can, of course, be modified, for example, to accommodate different subjects, subject response to a particular dosage schedule, different types of cancer, and / or different stages of cancer. In some implementations, the androgen receptor signaling inhibitor is selected from abiraterone, enzalutamide, apalutamide, and darolutamide. In some implementations, the compound administered to the subject is CBA-1 or CBA-2. In some implementations, the subject has prostate cancer. In some implementations, the compound administered is CBA-1 or CBA-2 and the androgen receptor signaling inhibitor is enzalutamide. In some implementations, the subject has CRPC, in some implementations, the subject has ARAT -resistant cancer. In some implementations, AR-V7 is expressed in a cancer cell present in the subject. Detection and / or diagnosis of prostate cancer, CRPC, ARAT -resistantcancer, and AR-V7 expression can be achieved utilizing the techniques described above with respect to the methods of inhibiting cancer cell proliferation.

[0042] As reflected in the discussion above, and as will be further evidenced in the discussion of the examples below, the compounds disclosed herein find utility in various methods for treating the human or animal body by therapy. In this regard, the compounds disclosed herein can find use in treating leukemia, lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, or combinations thereof.

[0043] Implementations of the various methods disclosed herein in which a combination including two or more of CBA-1, VMS-7-102, CBA-2, ZBC-1-82, and CBA-3 is administered to the subject are also contemplated herein.

[0044] It is appreciated that where reference is made to the use of a compound disclosed herein, embodiments and implementations utilizing a pharmaceutically acceptable salt of such compound are also contemplated.

[0045] The presently disclosed subject matter is further illustrated by the following specific but non-limiting examples. The following examples may include compilations of data that are representative of data gathered at various times during the course of development and experimentation related to the presently disclosed subject matter.EXAMPLES

[0046] The following examples relate to the detection of certain compounds’ ability to inhibit KDM3A activity and / or cancer cell proliferation in various types of cancers.

[0047] Materials and Methods for Examples 1-7

[0048] Materials. CBA-1, VMS-7-102, VMS-7-118 (CBA-2), ZBC-1-82, and ZBC-1-83(CBA-3) were synthesized. Exemplary reaction schemes for synthesizing ZBC-1-82, ZBC-1-83, and VMS-7-118 are shown in FIGS. 5, 6, and 7, respectively.

[0049] Chemicals were purchased from either Millipore Sigma (St. Louis, MO) or Fisher Scientific (Hampton, NH) unless otherwise specified. Solvents were used from commercial vendors without further purification unless otherwise noted. Nuclear magnetic resonance spectra were acquired on a Varian (XH at 400MHz;13C at 100MHz) instrument. High resolution electrospray ionization (ESI) mass spectra were recorded on an LTQ-Orbitrap Velos mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). The FT resolution was set at 100,000 (at 400 mlz). Samples were introduced through direct infusion using a syringe pump with a flow rate of 5pL / min. Compounds were chromatographed on preparative layer Merck silica gel F254 (Fisher Scientific) plates unless otherwise indicated.

[0050] KDM3A Activity Inhibition.

[0051] The inhibition of KDM3A enzymatic activity by V-7-118 (CBA-2) and ZBC-1-83 (CBA-3) was assessed through the services of BPS Bioscience (San Diego, California).

[0052] Cell Lines and Cell Culture. LS174T colon cancer cells and PC4, DU-145 and 22RV1 prostate cancer cells were cultured in RPMH640 (Sigma, R8758) containing 10% Fetal Bovine Serum (Sigma F0926). All cells were cultured at 37 U with 5% CO2 atmosphere in a water jacketed incubator (NuAire, Plymouth, MN).

[0053] Tumor Volume. Tumor volume in LS174T colorectal cancer (CRC) xenografted severe combined immunodeficient (SCID) mice treated with VMS-7-118 (CBA-2) (10 mg / kg twice per week, IP) and untreated LS174T colorectal cancer (CRC) xenografted controls over the course of 12 days was calculated based on the formula (length x width2) / 2 (four mice per group,two tumors on each mouse). Tumor volume in 22RV1 prostate cancer xenografted SCID mice treated with VMS-7-118 (CBA-2) (10 mg / kg, IP), 2RV1 prostate cancer xenografted SCID mice treated with enzalutamide (ENZ) (20 mg / kg per day, oral), and untreated 22RV1 prostate cancer xenografted controls over the course of 15 days was assessed via caliper and calculated based on the formula (length x width2) / 2.

[0054] Survival Rate of Treated LS174T CRC Xenografted Mice. The survival rate of LS174T CRC xenografted SCID mice treated with CBA-3 (2 mg / kg twice per week, OP, n =4) and untreated LS174T CRC xenografted control mice (n = 10) was assessed by log-rank (Mantel-Cox) test.

[0055] Cell Proliferation Inhibition Assays. All compounds to be tested were dissolved in DMSO at different concentrations. The cells were seeded into 12-well plates at a density of 40,000 cells per well in 1 mL of culture medium and were cultured overnight at 37 °C. Either 1 pL of the compounds or the vehicle control (DMSO) was added to the cells. For combination studies, enzalutamide (ENZ) was added together with CBA-1 or CBA-2 at the indicated concentrations. After 5 days, the medium was removed, and 200 pL of 0.25% trypsin was added. The cells were re-suspended in 800 pL phosphate-buffered saline (PBS) and counted with Vi- CELL XR 2.03 (Beckman Coulter, Inc. USA). The ratio R of the number of viable cells in the compound-treated group to the number of viable cells in the DMSO-treated group was called relative growth rate, and the growth inhibition was calculated as (l-R)*100.

[0056] H3K9Me2and Inhibition of Androgen Receptor (AR). AR-V7, and c-Myc. 22RV 1 cells were treated with DMSO, CBA-1, or CBA-2. The cell lysates were analyzed by western blot using antibodies that recognize H3K9Me2methylation, c-Myc, AR (both wild-type and AR-V7) (FIG. 9). P-Actin was analyzed as a loading control. The antibodies were purchased fromCell Signaling Technology.

[0057] NCI-60 Cell Line Screening. The NCI-60 cell lines were cultured in RPMI-1640 medium with 5% FBS. The cells were treated with 0.1 or 0.5 pM CBA-3, with DMSO as a control. After treatment (4-8 days depending on doubling time), the cells were counted with Vi- CELL XR 2.03 (Beckman Coulter, Inc. USA).

[0058] Example 1: Inhibition of KDM3A Activity

[0059] As shown in FIG. 1, VMS-7-118 (CBA-2) and ZBC-1-83 (CBA-3) were both found to significantly inhibit KDM3 A enzymatic activity as compared to the untreated controls at both tested concentrations (1 pM and 10 pM), with CBA-3 being more potent than CBA-2 at each concentration. Although the inhibitory effect of ZBC-1-82 against KDM3A enzymatic activity was not tested in this study it is believed that such compound is also likely to inhibit KDM3 A enzymatic activity because of its similarity in structure to CBA-3 and its ability, like CBA-3, to inhibit colon cancer cell growth (see TABLE 1). Although the inhibitory effects of VMS-7-102 against KDM3A enzymatic activity was not tested in this study it is believed that such compound is also likely to inhibit KDM3A enzymatic activity because of its similarity in structure to VMS- 7-118. It is appreciated that CBA-1 has also been found to inhibit KDM3A enzymatic activity.

[0060] Example 2: Inhibition of Cell Proliferation in Colon and Prostate Cancer Cell Lines

[0061] To assess the inhibitory efficacy of CBA-1, VMS-7-102, VMS-7-118 (CBA-2), ZBC- 1-82, and ZBC-1-83 (CBA-3) against colon cancer, and more particularly, CRC cell proliferation, CBA-1, VMS-7-102, VMS-7-118, ZBC-1-82, and ZBC-1-83 were each separately administered to LS174T colon cancer cell culture in 0.1 pM and 3 pM concentrations (TABLE 1). To assess the inhibitory efficacy of CBA-1, VMS-7-102, and VMS-7-118 against prostatecancer cell proliferation, and, in particular, castration resistant and androgen receptor axis- targeted therapy (ARAT)-resistant cancer, CBA-1, VMS-7-102, and VMS-7-118 were each separately administered to 22RV1 prostate cancer cell culture in 0.1 pM and 0.3 pM concentrations (TABLE 1). Cell proliferation was analyzed by Vi-Cell XR Cell Viability Analyzer (Beckman Coulter, Inc., USA).

[0062] Inhibition of CRC Cell Proliferation. As shown in TABLE 1, CBA-1, VMS-7-102, VMS-7-118, ZBC-1-82, and ZBC-1-83 were each found to inhibit CRC LS174T cell proliferation, with inhibition increasing as the concentration of each compound was increased. VMS-7-102 and VMS-7-118 were both found to provide greater inhibition of CRC cell proliferation than CBA-1 at both administered concentrations, which was surprising as CBA-1 has previously been found to provide greater Wnt signaling inhibition than VMS-7-102 and VMS-7-118. Likewise, ZBC-1-82 and ZBC-1-83 were found to provide greater inhibition of CRC cell proliferation than CBA at both administered concentrations, with ZBC-1-83 exhibiting the greatest inhibition against CRC cell proliferation at each concentration with 91% and 95.7% inhibition.TABLE 1. Inhibition of Colon Cancer and Prostate Cancer

[0063] Inhibition of Prostate Cancer Cell Proliferation. As further shown in TABLE 1, CBA- 1, VMS-7-102, and VMS-7-118 were each found to inhibit prostate cancer 22RV1 cell proliferation. Cell proliferation inhibition was generally found to increase with the application of such compounds at increased concentrations, with marked increases in inhibition being found in CBA-1 (0% to 57.7%) and VMS-7-118 (41.2% to 70.7%) being observed when the concentration of such compounds was increased from 0.1 pM to 0.3 pM.

[0064] Although the inhibitory effect of ZBC-1-82 and ZBC-1-83 against 22RV1 cell proliferation was not measured in this study, such compounds are nonetheless anticipated to inhibit 22RV1 cell proliferation. ZBC-1-83 was, as evidenced in Example 1, found to inhibit KDM3A enzymatic activity, which is overexpressed in and promotes growth of prostate cancer cells. As further discussed below in Example 6, ZBC-1-83 (CBA-3) has been found to inhibit cancer cell proliferation in other types of prostate cancer cells. The structural similarity between ZBC-1-82 and ZBC-1-83 suggests that ZBC-1-82 is also likely to inhibit prostate cancer cell proliferation, such as 22RV1.

[0065] Example 3: Inhibition of Colon Cancer Tumor Growth

[0066] To assess the inhibitory efficacy of VMS-7-118 against CRC tumor growth relative to untreated controls, VMS-7-118 (CBA-2) was administered (10 mg / kg, intraperitoneal (IP)) to LS174T colon cancer xenografted SCID mice. As shown in FIGS. 2A and 2B, significant decreases in tumor volume were observed in mice treated with VMS-7-118 as compared to untreated controls.

[0067] Although the effects of VMS-7-102, ZBC-1-82, and ZBC-1-83 against LS174T colon cancer tumor growth volume in SCID mice were not measured in the current study, it is believed that such compounds are also likely to inhibit tumor volume growth as these compounds have asimilar structure and mechanism (KDM3A inhibition) to VMS-7-118. Additionally, ZBC-1-82 and ZBC-1-83 were more active in inhibiting colon cancer cell growth in vitro, in at least some concentrations, than VMS-7-118 (TABLE 1), and ZBC-1-83 (CBA-3) has been found to increase the survival rate of LS174t CRC xenografted SCID mice, as further discussed below with respect to Example 4.

[0068] Example 4: Survival Rate of CBA-3-Treated CRC Mice

[0069] CBA-3 (ZBC-1-183) is an orally available analog of ZBC-1-82, suggesting that it has better pharmaceutical properties, including bioavailability.

[0070] As shown in FIG. 3, CBA-3 treatment (2 mg / kg, OP) significantly enhanced the survival rate of LS174T CRC xenografted SCID as compared to the untreated controls.

[0071] While CBA-1 was active in cell models, and previously found to be active in an in vivo zebrafish model, it was inactive in mouse-based models. Accordingly, the beneficial effects exhibited by VMS-7-118 (CBA-2) with respect to inhibiting tumor volume growth, and CBA-3 with respect to increasing the survival rate of CRC xenografted mice, may indicate that the structure of CBA-2 and CBA-3 better enables such compounds to be utilized in mammalian treatment applications. CBA-2 and CBA-3 have each been found to be more stable than in CBA- 1 in low pH buffer simulating gastric fluid (pH 1.5-3.5).

[0072] Example 5: Inhibition of Prostate Cancer Tumor Growth

[0073] To assess the inhibitory efficacy of VMS-7-118 (CBA-2) against CRPC tumor growth relative to untreated and enzalutamide (ENZ) controls, VMS-7-118 was administered to SCID mice with 22RV1 prostate cancer xenografts. As shown in FIGS. 4A and 4B, significant decreases in tumor volume was observed in mice treated with VMS-7-118 as compared to both untreated and ENZ-treated controls.

[0074] Although the inhibitory effect of VMS-7-102, ZBC-1-82, and ZBC-1-83 (CBA-3) against 22RV1 colon cancer tumor growth volume was not measured in the current study, it is believed that such compounds are also likely to inhibit tumor growth volume as these compounds have a similar structure and mechanism (KDM3A inhibition) to VMS-7-118. Additionally, CBA-3 has been found to inhibit cell proliferation in other types of prostate cancer cells, as further discussed below with respect to Example 6.

[0075] Example 6: Treatment of Cancer Resistant to Androgen Receptor Inhibitor Treatment

[0076] To overcome enzalutamide (ENZ) resistance, we screened a panel of commercial and home-made compounds, and identified a family of histone lysine demethylases 3 A (KDM3A) inhibitors that significantly enhance the therapeutic efficacy of enzalutamide on 22RV1 prostate cancer cells, which express the androgen receptor splice variant 7 (AR-V7). KDM3A regulates H3K9 methylation, which represses gene expression. Inhibition of KDM3A increases H3K9 methylation and blocks transcription. We originally developed KDM3A inhibitors to inhibit the Wnt / p-catenin signaling. We identified CBA-1 as an epigenetic regulator that inhibited KDMs possessing the Jumanji C (JmjC) domain. Based on structure-activity relationships (SAR) in conjunction with molecular docking studies, we identified CBA-2 (VMS-7-118) that inhibited 22RV1 prostate cancer xenografts growth in mouse models (FIG. 11).

[0077] KDM3A is overexpressed in prostate cancer and has been recognized as an important drug target for prostate cancer treatment. It is hypothesized that KDM3A inhibitors enhanced enzalutamide efficacy by multiple mechanisms: (1) inhibition of the expression of AR-V7; (2) inhibition of AR target gene expression; and (3) inhibition of other signaling pathways that contribute to enzalutamide resistance.

[0078] CBAs Inhibited Prostate Cancer Cells. CBA-1 was originally developed as a Wnt inhibitor for CRC treatment. CBA-2 and CBA-3 (ZBC-1-83) were subsequently designed and synthesized. CBA-3 was found to significantly inhibit prostate cancer cell lines PC3, an androgen independent cell line, and DU-145 (FIGS. 12 and 13). The inhibitory effects of CBA-1 and CBA-2 were assessed against the 22RV1 cell line, which is resistant to enzalutamide by expressing the AR-V7 variant. Both CBA-1 and CBA-2 were found to inhibit the proliferation of 22RV1 (FIGS. 10 and 11). Moreover, CBA-1 and CBA-2 significantly enhanced the efficacy of enzalutamide on 22RV1 cells (FIGS. 10 and 11), suggesting that KDM3A inhibitors can be used to treat enzalutamide-resistant and castration resistant prostate cancer, and that such inhibitors can be used in combination with enzalutamide. In this regard, treatment employing CBA-1 in combination with enzalutamide was found to provide greater 22RV1 cell proliferation inhibition than either CBA-1 or enzalutamide alone (FIG. 10). Similarly, treatment employing CBA-2 in combination with enzalutamide was found to provide greater 22RV1 cell proliferation inhibition than either CBA-2 or enzalutamide alone (FIG. 11).

[0079] CBAs Increased H3K9 Methylation and Inhibited AR-V7 Expression in 22Ryl Prostate Cancer Cells. Enzalutamide interacts with the ligand-binding domain (LBD) of full- length androgen receptor (AR) and inhibits AR signaling by blocking ligand binding, AR nuclear translocation and AR-medicated transcription (FIG. 8). However, 22RV1 cells are resistant to the AR-targeted therapy by expressing both full-length AR and the AR-V7 splicing variant. AR- V7 lacks LBD and activates transcription through ligand-independent mechanism. Since enzalutamide interacts with the LBD, it is ineffective to the AR-V7 variant. CBA-1 and CBA-2 were found to inhibit the expression of both full-length AR and AR-V7 (FIG. 9). It is hypothesized that H3K9 methylation regulates AR expression. In addition, CBA-1 and CBA-2were both found to repress the expression of c-Myc, an AR-target gene, suggesting that KDM3A also regulates AR-mediated transcription (FIG. 9).

[0080] Example 7: NCI-60 Cell Line Screening

[0081] KDM3A also regulates other cell signaling pathways and is an anti-cancer target for multiple cancers. Accordingly, to assess whether the compounds found effective with respect to colon cancer and prostate cancer cell proliferation may be useful in inhibiting other types of cancer cells, a NCI-60 (National Cancer Institute 60) cell line panel was carried out for ZBC-1- 83 (CBA-3) (TABLE 2).TABLE 2. NCI-60 Cell Proliferation Inhibition for ZBC-1-83.

[0082] As shown in TABLE 2, ZBC-1-83 provided significant inhibition of cancer cell proliferation for multiple cell lines for leukemia, lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer.

[0083] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference, including the references set forth in the following list:REFERENCES1. Liu, C.; Kato, Y.; Zhang, Z.; Do, V. M.; Yankner, B. A.; He, X., beta-Trcp couples beta- catenin phosphorylation-degradation and regulates Xenopus axis formation. Proc Natl AcadSci USA 1999, 96 (11), 6273-8.2. Liu, C.; Li, Y.; Semenov, M.; Han, C.; Baeg, G. H.; Tan, Y.; Zhang, Z.; Lin, X.; He, X., Control of beta-catenin phosphorylation / degradation by a dual-kinase mechanism. Cell 2002, 108 (6), 837-47.3. Shi, J.; Liu, Y.; Xu, X.; Zhang, W .; Yu, T.; Jia, J.; Liu, C., Deubiquitinase USP47 / UBP64E Regulates beta-Catenin Ubiquitination and Degradation and Plays a Positive Role in Wnt Signaling. Mol Cell Biol 2015, 35 (19), 3301-11.4. Yang, J.; Zhang, W.; Evans, P. M.; Chen, X.; He, X.; Liu, C., Adenomatous polyposis coli (APC) differentially regulates beta-catenin phosphorylation and ubiquitination in colon cancer cells. J Biol Chem 2006, 281 (26), 17751-7.5. Zhang, W.; Sviripa, V. M.; Xie, Y.; Yu, T.; Haney, M. G.; Blackbum, J. S.; Adeniran, C. A.; Zhan, C. G.; Watt, D. S.; Liu, C., Epigenetic Regulation of Wnt Signaling by Carboxamide-Substituted Benzhydryl Amines that Function as Histone Demethylase Inhibitors. iScience 2020, 23 (12), 101795.6. Zhang, W.; Sviripa, V.; Chen, X.; Shi, J.; Yu, T.; Hamza, A.; Ward, N. D.; Kril, L. M.; Vander Kooi, C. W.; Zhan, C. G.; Evers, B. M.; Watt, D. S.; Liu, C., Fluorinated N,N- dialkylaminostilbenes repress colon cancer by targeting methionine S- adenosyltransferase 2A. ACS Chem Biol 2013, 8 (4), 796-803.Giles, R. H.; van Es, J. H.; Clevers, H., Caught up in a Wnt storm: Wnt signaling in cancer. Biochim Biophys Acta 2003, 1653 (1), 1-24. Kinzler, K. W .; Vogelstein, B., Lessons from hereditary colorectal cancer. Cell 1996, 87 (2), 159-70. Polakis, P., Wnt signaling and cancer. Genes Dev 2000, 14 (15), 1837-51. Anastas, J. N.; Moon, R. T., WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer 2013, 13 (1), 11-26. Nusse, R ; Clevers, H., Wnt / beta-Catenin Signaling, Disease, and Emerging Therapeutic Modalities. Cell 2017, 169 (6), 985-999. Barker, N.; Clevers, EL, Mining the Wnt pathway for cancer therapeutics. Nat Rev Drug Discov 2006, 5 (12), 997-1014. Garber, K., Drugging the Wnt pathway: problems and progress. J Natl Cancer Inst 2009, 101 (8), 548-50. Zhong, Z.; Virshup, D. M., Wnt Signaling and Drug Resistance in Cancer. Mol Pharmacol 2020, 97 (2), 72-89. Chen, B.; Dodge, M. E.; Tang, W.; Lu, J.; Ma, Z.; Fan, C. W.; Wei, S.; Hao, W .; Kilgore, J.; Williams, N. S.; Roth, M. G.; Amatruda, J. F.; Chen, C.; Lum, L., Small molecule- mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer. Nat Chem Biol 2009, 5 (2), 100-7. Huang, S. M.; Mishina, Y. M.; Liu, S.; Cheung, A.; Stegmeier, F.; Michaud, G. A.; Charlat, O.; Wiellette, E.; Zhang, Y.; Wiessner, S.; Hild, M.; Shi, X.; Wilson, C. J.; Mickanin, C.; Myer, V.; Fazal, A.; Tomlinson, R.; Serluca, F.; Shao, W .; Cheng, H.; Shultz, M.; Rau, C.; Schirle, M.; Schlegl, J.; Ghidelli, S.; Fawell, S.; Lu, C.; Curtis, D.; Kirschner, M. W .; Lengauer, C.; Finan, P. M.; Tallarico, J. A.; Bouwmeester, T.; Porter, J. A.; Bauer, A.; Cong, F., Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling. Nature 2009. Liu, J.; Pan, S.; Hsieh, M. H.; Ng, N.; Sun, F.; Wang, T.; Kasibhatla, S.; Schuller, A. G.; Li, A. G.; Cheng, D.; Li, J.; Tompkins, C.; Pferdekamper, A.; Steffy, A.; Cheng, J.; Kowal, C.; Phung, V.; Guo, G.; Wang, Y.; Graham, M. P.; Flynn, S.; Brenner, J. C.; Li, C.; Villarroel, M. C.; Schultz, P. G.; Wu, X.; McNamara, P.; Sellers, W. R.; Petruzzelli, L.; Boral, A. L.; Seidel, H. M.; McLaughlin, M. E.; Che, J.; Carey, T. E.; Vanasse, G.;Harris, J. L., Targeting Wnt-driven cancer through the inhibition of Porcupine by LGK974. Proc Natl Acad Sci USA 2013, 110 (50), 20224-9. Lyou, Y.; Habowski, A. N.; Chen, G. T.; Waterman, M. L., Inhibition of nuclear Wnt signalling: challenges of an elusive target for cancer therapy. Br J Pharmacol 2017, 174 (24), 4589-4599. Lee, E.; Madar, A.; David, G.; Garabedian, M. J.; Dasgupta, R.; Logan, S. K., Inhibition of androgen receptor and beta-catenin activity in prostate cancer. Proc Natl Acad Sci U S A 2013, 110 (39), 15710-5. Lepourcelet, M.; Chen, Y. N.; France, D. S.; Wang, H.; Crews, P.; Petersen, F.; Bruseo,C.; Wood, A. W .; Shivdasani, R. A., Small-molecule antagonists of the oncogenic Tcf / beta-catenin protein complex. Cancer Cell 2004, 5 (1), 91-102. Schneider, J. A.; Craven, T. W .; Kasper, A. C.; Yun, C.; Haugbro, M.; Briggs, E. M.; Svetlov, V.; Nudler, E.; Knaut, H.; Bonneau, R.; Garabedian, M. J.; Kirshenbaum, K.; Logan, S. K., Design of Peptoid-peptide Macrocycles to Inhibit the beta-catenin TCF Interaction in Prostate Cancer. Nat Commun 2018, 9 (1), 4396. Feng, M.; Jin, J. Q.; Xia, L.; Xiao, T.; Mei, S.; Wang, X.; Huang, X.; Chen, J.; Liu, M.; Chen, C.; Rafi, S.; Zhu, A. X.; Feng, Y. X.; Zhu, D., Pharmacological inhibition of beta- catenin / BCL9 interaction overcomes resistance to immune checkpoint blockades by modulating Treg cells. Sci Adv 2019, 5 (5), eaau5240. Wisniewski, J. A.; Yin, J.; Teuscher, K. B.; Zhang, M.; Ji, H., Structure-Based Design of 1,4-Dibenzoylpiperazines as beta-Catenin / B-Cell Lymphoma 9 Protein-Protein Interaction Inhibitors. ACS Med Chem Lett 2016, 7 (5), 508-13. Emami, K. H.; Nguyen, C.; Ma, H.; Kim, D. H.; Jeong, K. W .; Eguchi, M.; Moon, R. T.; Teo, J. L.; Kim, H. Y.; Moon, S. H.; Ha, J. R.; Kahn, M., A small molecule inhibitor of beta-catenin / CREB-binding protein transcription [corrected]. Proc Natl Acad Sci USA 2004, 101 (34), 12682-7. Lenz, H. J.; Kahn, M., Safely targeting cancer stem cells via selective catenin coactivator antagonism. Cancer Sci 2014, 105 (9), 1087-92. Zhang, W.; Sviripa, V. M.; Kril, L. M.; Yu, T.; Xie, Y.; Hubbard, W. B.; Sullivan, P. G.; Chen, X.; Zhan, C. G.; Yang-Hartwich, Y.; Evers, B. M.; Spear, B. T.; Gedaly, R.; Watt,D. S.; Liu, C., An Underlying Mechanism of Dual Wnt Inhibition and AMPK Activation:Mitochondrial Uncouplers Masquerading as Wnt Inhibitors. J Med Chem 2019, 62 (24), 11348-11358. Sierra, J.; Yoshida, T.; Joazeiro, C. A.; Jones, K. A., The APC tumor suppressor counteracts beta-catenin activation and H3K4 methylation at Wnt target genes. Genes & development 2006, 20 (5), 586-600. Cloos, P. A.; Christensen, J.; Agger, K.; Helin, K., Erasing the methyl mark: histone demethylases at the center of cellular differentiation and disease. Genes & development 2008, 22 (9), 1115-40. Jambhekar, A.; Anastas, J. N.; Shi, Y., Histone Lysine Demethylase Inhibitors. Cold Spring Harb Per spect Med 2017, 7 (1). Klose, R. J.; Kallin, E. M.; Zhang, Y., JmjC-domain-containing proteins and histone demethylation. Nat Rev Genet 2006, 7 (9), 715-27. Shi, Y.; Lan, F.; Matson, C.; Mulligan, P.; Whetstine, J. R.; Cole, P. A.; Casero, R. A.; Shi, Y , Histone demethylation mediated by the nuclear amine oxidase homolog LSD1. Cell 2004, 119 (7), 941-53. Kooistra, S. M.; Helin, K., Molecular mechanisms and potential functions of histone demethylases. Nat Rev Mol Cell Biol 2012, 13 (5), 297-311. Sviripa, V. M.; Zhang, W.; Balia, A. G.; Tsodikov, O. V.; Nickell, J. R.; Gizard, F.; Yu, T.; Lee, E. Y.; Dwoskin, L. P.; Liu, C.; Watt, D. S., 2',6'-Dihalostyrylanilines, pyridines, and pyrimidines for the inhibition of the catalytic subunit of methionine S- adenosyltransferase-2. JMedChem 2014, 57 (14), 6083-91. MacDonald, B. T.; Tamai, K.; He, X., Wnt / beta-Catenin Signaling: Components, Mechanisms, and Diseases. Developmental Cell 2009, 17 (1), 9-26. Comprehensive molecular characterization of human colon and rectal cancer. Nature 2012, 487 (7407), 330-7. Schatoff, E. M.; Leach, B. I.; Dow, L. E., Wnt Signaling and Colorectal Cancer. Current colorectal cancer reports 2017, 13 (2), 101-110. Patnaik, S.; Anupriya, Drugs Targeting Epigenetic Modifications and Plausible Therapeutic Strategies Against Colorectal Cancer. Front Pharmacol 2019, 10, 588. Polakis, P., Wnt signaling in cancer. Cold Spring Harb Perspect Biol 2012, 4 (5).Strekalova, E.; Malin, D.; Weisenhorn, E. M. M.; Russell, J. D ; Hoelper, D.; Jain, A.;Coon, J. J.; Lewis, P. W.; Cryns, V. L., S-adenosylmethionine biosynthesis is a targetable metabolic vulnerability of cancer stem cells. Breast Cancer Res Treat 2019, 175 (1), 39- 50. Zhang, W .; Sviripa, V.; Kril, L. M.; Chen, X.; Yu, T.; Shi, J.; Rychahou, P.; Evers, B. M.; Watt, D. S.; Liu, C., Fluorinated N,N-dialkylaminostilbenes for Wnt pathway inhibition and colon cancer repression. J Med Chem 2011, 54 (5), 1288-97. Zaytseva, Y. Y.; Rychahou, P. G.; Le, A. T.; Scott, T. L.; Flight, R. M.; Kim, J. T.; Harris, J.; Liu, J.; Wang, C.; Morris, A. J.; Sivakumaran, T. A.; Fan, T.; Moseley, H.; Gao, T.; Lee, E. Y.; Weiss, H. L.; Heuer, T. S.; Kemble, G.; Evers, M., Preclinical evaluation of novel fatty acid synthase inhibitors in primary colorectal cancer cells and a patient-derived xenograft model of colorectal cancer. Oncotarget 2018, 9 (37), 24787- 24800. Hedgepeth, C. M.; Conrad, L. J.; Zhang, J.; Huang, H. C.; Lee, V. M.; Klein, P. S., Activation of the Wnt signaling pathway: a molecular mechanism for lithium action. Dev Biol ' 1997, 755 (1), 82-91. Peng, K.; Su, G.; Ji, J.; Yang, X.; Miao, M.; Mo, P.; Li, M.; Xu, J.; Li, W .; Yu, C , Histone demethylase JMJD1A promotes colorectal cancer growth and metastasis by enhancing Wnt / beta-catenin signaling. J Biol Chem 2018, 293 (27), 10606-10619. Liu, J.; Liang, T.; Zhangsun, W ., KDM3A is associated with tumor metastasis and modulates colorectal cancer cell migration and invasion. Int J Biol Macromol 2019 , 126, 318-325. Li, J.; Yu, B.; Deng, P.; Cheng, Y.; Yu, Y.; Kevork, K.; Ramadoss, S.; Ding, X.; Li, X.; Wang, C. Y., KDM3 epigenetically controls tumorigenic potentials of human colorectal cancer stem cells through Wnt / beta-catenin signalling. Nat Commun 2017, 8, 15146. Wang, H. Y.; Long, Q. Y ; Tang, S. B.; Xiao, Q.; Gao, C.; Zhao, Q. Y.; Li, Q. L ; Ye, M.;Zhang, L.; Li, L. Y.; Wu, M., Histone demethylase KDM3A is required for enhancer activation of hippo target genes in colorectal cancer. Nucleic Acids Res 2019, 47 (5), 2349-2364. Evans, P. M.; Chen, X.; Zhang, W .; Liu, C., KLF4 interacts with beta-catenin / TCF4 and blocks p300 / CBP recruitment by beta-catenin. Mol Cell Biol 2010, 30 (2), 372-81.Evans, P. M.; Zhang, W.; Chen, X.; Yang, J.; Bhakat, K. K.; Liu, C., Kruppel-like factor 4 is acetylated by p300 and regulates gene transcription via modulation of histone acetylation. J Biol Chem 2007, 282 (47), 33994-4002. Yu, T.; Chen, X.; Zhang, W.; Colon, D.; Shi, J.; Napier, D.; Rychahou, P.; Lu, W.; Lee, E. Y.; Weiss, H. L.; Evers, B. M.; Liu, C., Regulation of the potential marker for intestinal cells, Bmil, by beta-catenin and the zinc finger protein KLF4: implications for colon cancer. J Biol Chem 2012, 287 (6), 3760-8. Burikhanov, R.; Sviripa, V. M.; Hebbar, N.; Zhang, W .; Layton, W. J.; Hamza, A.; Zhan, C. G.; Watt, D. S.; Liu, C.; Rangnekar, V. M., Arylquins target vimentin to trigger Par-4 secretion for tumor cell apoptosis. Nat Chem Biol 2014, 10 (11), 924-926. Frasinyuk, M. S.; Zhang, W.; Wyrebek, P.; Yu, T.; Xu, X.; Sviripa, V. M.; Bondarenko, S. P.; Xie, Y.; Ngo, H. X.; Morris, A. J.; Mohler, J. L.; Fiandalo, M. V.; Watt, D. S.; Liu, C., Developing antineoplastic agents that target peroxisomal enzymes: cytisine-linked isoflavonoids as inhibitors of hydroxy steroid 17-beta-dehydrogenase-4 (HSD17B4). Org Biomol Chem 2017, 15 (36), 7623-7629. Berg, K. C. G.; Eide, P. W .; Eilertsen, I. A.; Johannessen, B.; Bruun, J.; Danielsen, S. A.; Bjornslett, M.; Meza-Zepeda, L. A.; Eknaes, M.; Lind, G. E.; Myklebost, O.; Skotheim, R. I.; Sveen, A.; Lothe, R. A., Multi-omics of 34 colorectal cancer cell lines - a resource for biomedical studies. Mol Cancer 2017, 16 (1), 116. Barretina, J.; Caponigro, G.; Stransky, N.; Venkatesan, K.; Margolin, A. A.; Kim, S.; Wilson, C. J.; Lehar, J.; Kryukov, G. V.; Sonkin, D.; Reddy, A.; Liu, M.; Murray, L.; Berger, M. F.; Monahan, J. E.; Morais, P ; Meltzer, J.; Korejwa, A.; Jane-Valbuena, J.; Mapa, F. A.; Thibault, J.; Bric-Furlong, E.; Raman, P.; Shipway, A.; Engels, I. H.; Cheng, J.; Yu, G. K.; Yu, J.; Aspesi, P., Jr.; de Silva, M.; Jagtap, K.; Jones, M. D.; Wang, L.; Hatton, C.; Palescandolo, E.; Gupta, S.; Mahan, S.; Sougnez, C.; Onofrio, R. C.; Liefeld, T.; MacConaill, L.; Winckler, W .; Reich, M.; Li, N.; Mesirov, J. P.; Gabriel, S. B.; Getz, G.; Ardlie, K.; Chan, V.; Myer, V. E.; Weber, B. L.; Porter, J.; Warmuth, M.; Finan, P.; Harris, J. L.; Meyerson, M.; Golub, T. R.; Morrissey, M. P.; Sellers, W. R.; Schlegel, R.; Garraway, L. A., The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 2012, 483 (7391), 603-7.54. Wen, Y. A.; Xing, X.; Harris, J. W.; Zaytseva, Y. Y.; Mitov, M. I.; Napier, D. L.; Weiss, H. L.; Mark Evers, B.; Gao, T., Adipocytes activate mitochondrial fatty acid oxidation and autophagy to promote tumor growth in colon cancer. Cell Death Dis 2017, 8 (2), e2593.55. Veschi, S.; De Lellis, L.; Florio, R ; Lanuti, P.; Massucci, A.; Tinari, N.; De Tursi, M.; di Sebastiano, P.; Marchisio, M.; Natoli, C.; Cama, A., Effects of repurposed drug candidates nitroxoline and nelfinavir as single agents or in combination with erlotinib in pancreatic cancer cells. J Exp Clin Cancer Res 2018, 37 (1), 236.56. Xie, Y.; Kril, L. M.; Yu, T.; Zhang, W .; Frasinyuk, M. S.; Bondarenko, S. P.;Kondratyuk, K. M.; Hausman, E.; Martin, Z. M.; Wyrebek, P. P.; Liu, X.; Deaciuc, A.; Dwoskin, L. P.; Chen, J.; Zhu, H.; Zhan, C. G.; Sviripa, V. M.; Blackburn, J.; Watt, D. S.; Liu, C., Semisynthetic aurones inhibit tubulin polymerization at the colchicine- binding site and repress PC -3 tumor xenografts in nude mice and myc-induced T-ALL in zebrafish. Sci Rep 2019, 9 (1), 6439.57. Wen, Y. A.; Xiong, X.; Zaytseva, Y. Y.; Napier, D. L.; Vallee, E.; Li, A. T.; Wang, C.; Weiss, H. L.; Evers, B. M.; Gao, T., Downregulation of SREBP inhibits tumor growth and initiation by altering cellular metabolism in colon cancer. Cell death & disease 2018, 9 (3), 265.58. Kondo, J.; Endo, H.; Okuyama, H.; Ishikawa, O.; lishi, H.; Tsujii, M.; Ohue, M.; Inoue, M., Retaining cell-cell contact enables preparation and culture of spheroids composed of pure primary cancer cells from colorectal cancer. Proc Natl Acad Sci USA 2011, 108 (15), 6235-40.59. U.S. Patent Application Publication No. 2023 / 0108479, entitled “Substituted N- Benzhydrylacetamide Inhibitors of Jamanji Domain Histone Demethylases for the Treatment of Cancer”, published April 6, 2023.

[0084] This disclosure further encompasses the following aspects.

[0085] Aspect 1. A method for inhibiting cancer cell proliferation in a subject, comprising administering to the subject a compound selected from the group consisting of:

[0086] Aspect 2. The method of aspect 1, wherein the subject has a cancer selected from the group consisting of: leukemia, lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer; and combinations thereof.

[0087] Aspect 3. The method of aspect 1, wherein the subject has prostate cancer.

[0088] Aspect 4. The method of aspect 3, wherein the subject has castration-resistant prostate cancer.

[0089] Aspect 5. The method of any of aspects 1 to 4, wherein the subject has androgen receptor axis-targeted therapy (ARAT)-resistant cancer.

[0090] Aspect 6. The method of any of aspects 1 to 5, wherein androgen receptor splice variant 7 (AR-V7) is expressed in a cancer cell present in the subject.

[0091] Aspect 7. The method of aspect 6, and further comprising detecting AR-V7 expression in the cancer cell present in the subject.

[0092] Aspect 8. The method of any of aspects 1 to 7, and further comprising administering an androgen receptor signaling inhibitor to the subject.

[0093] Aspect 9. The method of aspect 8, wherein the androgen receptor signaling inhibitor is enzalutamide.

[0094] Aspect 10. The method of aspect 9, wherein the compound is

[0095] Aspect 11. The method of any of aspects 1 to 10, wherein the subject has colon cancer.

[0096] Aspect 12. The method of aspect 11, wherein the compound is

[0097] Aspect 13. The method of any of aspects 1 to 12, wherein the subject is a mammal.

[0098] Aspect 14. A compound for inhibiting cancer cell proliferation, comprising a structure according to Formula I:wherein R1is heterocycle.

[0099] Aspect 15. The compound of aspect 14, wherein the compound is

[0100] Aspect 16. A method for improving the therapeutic effect of an androgen receptor signaling inhibitor, comprising administering to a subject with cancer a compound selected from the group consisting of:

[0101] Aspect 17. The method of aspect 16, wherein the subject has prostate cancer.

[0102] Aspect 18. The method of aspect 16 or 17, wherein the subject has castrationresistant prostate cancer.

[0103] Aspect 19. The method of any of aspects 16 to 18, wherein the subject has androgen receptor axis-targeted therapy (ARAT)-resistant cancer.

[0104] Aspect 20. The method of any of aspects 16 to 19, wherein androgen receptor splice variant 7 (AR-V7) is expressed in a cancer cell present in the subject.

[0105] Aspect 21. The method of any of claims 16 to 20, wherein the androgen receptor signaling inhibitor is enzalutamide.

[0106] Aspect 22. A compound having a structureuse in a method of treating the human or animal body by therapy.

[0107] Aspect 23. A compound having a structurea method of treating the human or animal body by therapy.

[0108] Aspect 24. A compound having a structurer use in a method of treating the human or animal body by therapy.

[0109] Aspect 25. A compound having a structurea method of treating the human or animal body by therapy.

[0110] Aspect 26. A compound having a structureuse in treating prostate cancer.

[0111] Aspect 27. A compound having a structurer use in treating prostate cancer.

[0112] Aspect 28. A compound having a structuretreating prostate cancer.

[0113] Aspect 29. A compound having a structureuse in treating prostate cancer.

[0114] Aspect 30. The compound of any of aspects 26 to 29, wherein the prostate cancer is castration-resistant prostate cancer.

[0115] Aspect 31. The compound of any of aspects 26 to 30, wherein the prostate cancer is ARAT-resistant cancer.

[0116] Aspect 32. The compound of any of aspects 26 to 31, wherein the prostate cancer is cancer which includes a cancer cell in which androgen receptor splice variant 7 (AR-V7) is expressed.

[0117] Aspect 33. A compound having a structureuse in treating colon cancer.

[0118] Aspect 34. A compound having a structuretreating colon cancer.

[0119] Aspect 35. A compound having a structurer use in treating colon cancer.

[0120] Aspect 36. A compound having a structuretreating colon cancer.

[0121] Aspect 37. A compound having a structureuse in treating colon cancer.

[0122] Aspect 38. A compound having a structureuse in treating a cancer selected from leukemia, lung cancer, central nervous system, melanoma, ovarian, renal, or breast cancer.

[0123] It will be understood that various details of the presently disclosed subject matter can be changed without departing from the scope of the subject matter disclosed herein. Furthermore, the foregoing description is for the purpose of illustration only, and not for the purpose of limitation.

Claims

CLAIMSWhat is claimed is:

1. A method for inhibiting cancer cell proliferation in a subject, comprising administering to2. The method of claim 1, wherein the subject has a cancer selected from the group consisting of: leukemia, lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer; and combinations thereof.

3. The method of claim 1, wherein the subject has prostate cancer.

4. The method of claim 3, wherein the subject has castration-resistant prostate cancer.

5. The method of claim 1, wherein the subject has androgen receptor axis-targeted therapy (ARAT)-resistant cancer.

6. The method of claim 1, wherein androgen receptor splice variant 7 (AR-V7) is expressed in a cancer cell present in the subject.

7. The method of claim 6, and further comprising detecting AR-V7 expression in the cancer cell present in the subject.

8. The method of claim 1, and further comprising administering an androgen receptor signaling inhibitor to the subject.

9. The method of claim 8, wherein the androgen receptor signaling inhibitor is enzalutamide.

10. The method of claim 9, wherein the compound i11. The method of claim 1, wherein the subject has colon cancer.m 11, wherein the compound i13. The method of any of claim 1, wherein the subject is a mammal.

14. A compound for inhibiting cancer cell proliferation, comprising a structure according toFormula I:wherein R1is heterocycle.

15. The compound of claim 14, wherein the compound is16. A method for improving the therapeutic effect of an androgen receptor signaling inhibitor, comprising administering to a subject with cancer a compound selected from the group consisting of:

17. The method of claim 16, wherein the subject has prostate cancer.

18. The method of claim 17, wherein the subject has castration-resistant prostate cancer.

19. The method of claim 16, wherein the subject has androgen receptor axis-targeted therapy (ARAT)-resistant cancer.

20. The method of any of claim 16, wherein androgen receptor splice variant 7 (AR-V7) is expressed in a cancer cell present in the subject.

21. The method of claim 16, wherein the androgen receptor signaling inhibitor is enzalutamide.

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