CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63/303,341 , filed January 26, 2022, and U.S. Provisional Application No. 63/330,878, filed April 14, 2022, the contents of which are incorporated herein by reference in their entirety.

FIELD

[0002] The present disclosure relates generally to methods, uses and kits for treating prostate cancer using combinations of cepharanthine and a taxane. In particular, the present invention pertains to the use of a combination of cepharanthine and a taxane such as cabazitaxel, docetaxel, or paclitaxel, for the treatment of prostate cancer.

INTRODUCTION

[0003] According to the American Cancer Society, in 2021 there were approximately 248,530 new cases of prostate cancer and about 34,130 deaths from prostate cancer in the United States, and it is the second leading cause of cancer death in American men. Metastatic prostate cancer is commonly treated using androgen deprivation therapy (ADT), including surgical or medical castration, but metastatic castration-resistant prostate cancer (mCRPC) commonly emerges (reviewed in e.g. Komura et al. 2019). The taxanes docetaxel and cabazitaxel are approved for the treatment of mCRPC, however, docetaxel resistance is known to develop.

[0004] Cepharanthine is a natural alkaloid extracted from the plant Stephania cepharantha Hayata. Cepharanthine has been widely used in Japan since the 1950s to treat both acute and chronic diseases, such as radiation-induced leukopenia, venomous snake bites, alopecia and malaria (see e.g. Bailly, 2019; Rogosnitzky & Danks, 2011).

[0005] Studies demonstrate the efficacy of cepharanthine and cepharanthine hydrochloride in the treatment of various cancers. Specifically, cepharanthine has been studied for its activity as a potential anti-tumor agent, reverse multi-drug resistance, and potentiate chemotherapy (see e.g. Bailly, 2019; Rogosnitzky & Danks, 2011). Cepharanthine is known to sensitize prostate cancer cell lines DU-145 and LNCaP to onconase cytotoxicity (Ita et al. 2008).

[0006] Cepharanthine is available in tablet and powder form for oral administration as well as in an injectable form. However, the alkaloid free base form of cepharanthine has very limited solubility (< 1 mg/mL). To overcome the poor solubility US patent No. 10,576,077 discloses the preparation of a pharmaceutically acceptable salt form of cepharanthine, cepharanthine.2HCI, and enteric formulations of cepharanthine.2HCI, as well as use of such cepharanthine.2HCI salts for the treatment of filovirus infections such as Ebola and Marburg viruses and subtypes thereof, other viral infections, hypertension, cancer, and neutropenia.

SUMMARY

[0007] As shown herein, it has been found that cepharanthine and the taxane cabazitaxel act synergistically to inhibit the growth of prostate cancer cells.

[0008] Accordingly, an aspect of the present disclosure includes a method of treating prostate cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a combination of cepharanthine, or pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof.

[0009] Another aspect includes a method for inhibiting the growth of a prostate cancer cell, the method comprising contacting the prostate cancer cell with a combination of cepharanthine or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof.

[0010] A further aspect includes a use of a combination of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof, for the treatment of prostate cancer.

[0011] Another aspect includes use of a combination of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof, for inhibiting the growth of a prostate cancer cell.

[0012] Another aspect includes a use of a combination of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof, in the manufacture of a medicament for the treatment of prostate cancer in a subject in need thereof. [0013] Another aspect includes a use of a combination of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof, in the manufacture of a medicament for inhibiting the growth of a prostate cancer cell.

[0014] Another aspect includes a combination of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof, for use in the treatment of prostate cancer in a subject in need thereof.

[0015] A further aspect includes a combination of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof, for use in inhibiting the growth of a prostate cancer cell.

[0016] Other aspects of the disclosure include combination therapies, compositions, and/or kits comprising cepharanthine or a pharmaceutical salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof, for use in the treatment of prostate cancer or inhibiting the growth of a prostate cancer cell.

[0017] In some embodiments of the various aspects described herein, the taxane is paclitaxel, docetaxel, or cabazitaxel. In some embodiments, the taxane is cabazitaxel.

[0018] In some embodiments of the various aspects described herein, cabazitaxel is administered, formulated for administration, or for use as a 1-hour intravenous infusion of about 20 mg/m ² of cabazitaxel every 3 weeks.

[0019] In some embodiments of the various aspects described herein, the cepharanthine salt is cepharanthine.2HCI.

[0020] In some embodiments of the various aspects described herein, the cepharanthine or pharmaceutically acceptable salt or solvate thereof is administered, formulated for administration, or for use at a dose between about 5 mg/day to about 400 mg/day, optionally about 10 mg/day, about 15 mg/day, about 20 mg/day, about 25 mg/day, about 50 mg/day, about 100 mg/day, about 150 mg/day, about 200 mg/day, about 300 mg/day, or about 400 mg/day.

[0021] In some embodiments of the various aspects described herein, the cepharanthine or pharmaceutically acceptable salt or solvate thereof is administered, formulated for administration, or for use at a daily dose between about 0.1 mg/kg body weight to about 4 mg/kg body weight, optionally about 0.244 mg/kg body weight, about 0.732 mg/kg body weight, or about 2.195 mg/kg body weight.

[0022] In some embodiments of the various aspects described herein, the prostate cancer is relapsed prostate cancer and/or has acquired chemoresistance.

[0023] In some embodiments of the various aspects described herein, the prostate cancer is a castration-resistant prostate cancer (CRPC).

[0024] In some embodiments of the various aspects described herein, the prostate cancer is metastatic prostate cancer.

[0025] In some embodiments of the various aspects described herein, the cepharanthine or pharmaceutically acceptable salt or solvate thereof and taxane or pharmaceutically acceptable solvate thereof are administered, formulated for administration, or for use simultaneously or sequentially.

[0026] In some embodiments of the various aspects described herein, the cepharanthine or pharmaceutically acceptable salt or solvate thereof is administered, formulated for administration, or for use orally. In some embodiments, the cepharanthine.2HCI is formulated as an enteric formulation for oral administration. In some embodiments, the enteric formulation for oral administration comprises about 25% w/w cepharanthine.2HCI in about 75% w/w Eudragit L100.

DRAWINGS

[0027] Further objects, features and advantages of the disclosure will become apparent from the following detailed description taken in conjunction with the accompanying figures showing illustrative embodiments of the disclosure, in which:

[0028] Fig. 1 shows the dose response curves for cepharanthine.2HCI, cisplatin, and cabazitaxel in DU 145 cells.

[0029] Fig. 2 shows the dose response curves for cepharanthine.2HCI, cisplatin, and cabazitaxel in 22Rv1 cells.

[0030] Fig. 3 shows the dose response curves for cepharanthine.2HCI, cisplatin, and cabazitaxel in LNCaP clone FGC cells.

[0031] Fig. 4A - Fig. 4E show the combination effect of cepharanthine.2HCL and cabazitaxel on 22Rv1 cells. (A) Dose response curve for cepharanthine.2HCL. (B) Dose response curve for cabazitaxel. (C) Dose response curve for cepharanthine.2HCL + cabazitaxel combination. (D) Combination Index plot. (E) Curve shift plot.

[0032] Fig. 5A - Fig. 5C show the combination effect of cepharanthine and cabazitaxel in an in vivo tumor model. The treatment was performed for 21 days. The study was terminated on Day 28. (A) The mean tumor volume curve of vehicle and treatment groups with SEM. (B) The mean tumor volume percentage curves of treatment vs vehicle groups. (C) The mean tumor volume growth inhibition percentage curves of treatment vs vehicle groups.

[0033] Fig. 6A - Fig. 6B show the body weight change percentage curves for in vivo mouse tumor model in Example 3. (A) Mean body weight % change ± SEM of vehicle and treatment groups. (B) Mean body weight ± SEM from group 5, 6, 7 and 8.

[0034] Fig. 7 shows survival curves of all groups at day 28 termination of study.

DESCRIPTION OF VARIOUS EMBODIMENTS

[0035] The following is a detailed description provided to aid those skilled in the art in practicing the present disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The terminology used in the description herein is for describing particular embodiments only and is not intended to be limiting of the disclosure. All publications, patent applications, patents, figures and other references mentioned herein are expressly incorporated by reference in their entirety.

[0036] Further, the definitions and embodiments described in particular sections are intended to be applicable to other embodiments herein described for which they are suitable as would be understood by a person skilled in the art. For example, in the following passages, different aspects of the disclosure are defined in more detail. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature described herein may be combined with any other feature or features described herein.

I. Definitions

[0037] As used herein, the following terms may have meanings ascribed to them below. However, it should be understood that other meanings that are known or understood by those having ordinary skill in the art are also possible, and within the scope of the present disclosure. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0038] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the description. Ranges from any lower limit to any upper limit are contemplated.

[0039] The term “about” as used herein may be used to take into account experimental error and variations that would be expected by a person having ordinary skill in the art. For example, “about” may mean plus or minus 10%, or plus or minus 5% of the indicated value to which reference is being made.

[0040] As used herein the singular forms "a", "an", and "the" include plural references unless the context clearly dictates otherwise.

[0041] The phrase "and/or," as used herein, should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with "and/or" should be construed in the same fashion, i.e., "one or more" of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the "and/or" clause, whether related or unrelated to those elements specifically identified.

[0042] As used herein in the specification and in the claims, "or" should be understood to have the same meaning as "and/or" as defined above. For example, when separating items in a list, "or" or "and/or" shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as "only one of' or "exactly one of" or, when used in the claims, "consisting of' will refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or" as used herein shall only be interpreted as indicating exclusive alternatives (i.e., "one or the other but not both") when preceded by terms of exclusivity, such as "either," "one of," "only one of," or "exactly one of." [0043] As used herein, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like are to be understood to be open-ended, i.e. , to mean including but not limited to. Only the transitional phrases "consisting of” and "consisting essentially of” shall be closed or semi-closed transitional phrases, respectively

[0044] As used herein, the phrase "at least one," in reference to a list of one or more elements, should be understood to mean at least one element selected from anyone or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified.

[0045] It should also be understood that, in certain methods described herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited unless the context indicates otherwise.

II. Methods and Uses

[0046] As shown herein, both cepharanthine and the taxane cabazitaxel act to inhibit the growth of prostate cancer cells. Unexpectedly it has been found that cepharanthine and cabazitaxel act synergistically to inhibit the growth of prostate cancer cells. Accordingly, an aspect of the present disclosure is a method of treating prostate cancer comprising administering to a subject in need thereof a therapeutically effective amount of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane or pharmaceutically acceptable solvate thereof. Another aspect of the disclosure includes a method of inhibiting growth of a prostate cancer cell, the method comprising contacting the prostate cancer cell with a combination of cepharanthine or a pharmaceutically acceptable salt or solvate thereof and a taxane or pharmaceutically acceptable solvate thereof. A further aspect is a combination therapy comprising cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane, or pharmaceutically acceptable solvate thereof, for use in treating prostate cancer. Another aspect includes a combination therapy comprising cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane, or pharmaceutically acceptable solvate thereof, for use in inhibiting growth of a prostate cancer cell. Also provided herein is a use of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane, or pharmaceutically acceptable solvate thereof, for the treatment of prostate cancer in a subject in need thereof. A further aspect is a use of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane, or pharmaceutically acceptable solvate thereof, for inhibiting growth of a prostate cancer cell. Also provided herein is a use of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane, or pharmaceutically acceptable solvate thereof, in the manufacture of a medicament for the treatment of prostate cancer in a subject in need thereof. Another aspect includes a use of cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, and a taxane, or pharmaceutically acceptable solvate thereof, in the manufacture of a medicament for inhibiting growth of a prostate cancer cell.

[0047] In an embodiment the taxane is cabazitaxel, paclitaxel, or docetaxel.

[0048] In an embodiment, the taxane is cabazitaxel.

[0049] The term “cepharanthine” refers to the compound with the following structure: or a pharmaceutically acceptable salt or solvate thereof. Pharmaceutically acceptable salts of cepharanthine may include, for example, salts prepared using an organic or inorganic acid such as hydrochloric, formic, maleic, benzoic, hydrobromic, acetic, malonic, glucuronic, phosphoric, methanesulfonic, glutamic, fumaric, sulfuric, tosic, aspartic, nitric, tartaric, lactic, pyruvic, or mucic acid. In an embodiment, the cepharanthine salt is prepared using hydrochloric acid. [0050] In an embodiment, the cepharanthine is a pharmaceutically acceptable salt such as, for example, cepharanthine»2HCI salt having the structure:

[0051] In an embodiment, the cepharanthine, or pharmaceutically acceptable salt or solvate thereof, is prepared in various forms and/or polymorphs, including amorphous phase, crystalline forms, milled forms and nano-particulate forms, and/or mixtures thereof.

[0052] In an embodiment the cepharanthine is enterically formulated. Enterically formulated salts of cepharanthine are disclosed for example in United States Patent No. 10,576,077, herein incorporated by reference in its entirety. Optionally, the enterically formulated cepharanthine.2HCI salt comprises about 25% cepharanthine.2HCI in Eudragit L100 (1/1 :methacrylic acid/methyl methacrylate enteric copolymer).

[0053] The term “taxane” as used herein refers to a family of diterpene compounds, or pharmaceutically acceptable solvates thereof, originally isolated from plants of the genus Taxus, and which act as microtubule poisons by stabilizing GDP- bound tubulin and inhibiting microtubule depolymerization. Common taxanes which are widely used for chemotherapy include paclitaxel (Taxol™), docetaxel (Taxotere™), and cabazitaxel (Jevtana™).

[0054] As used herein, “solvate” means a solvation product of a compound, formed for example as a combination of solvent molecules, such as water, an alcohol, or other polar organic solvent, with molecules or ions of the solute compound. Suitable solvents are physiologically tolerable at the dosage administered, and may include, without limitation, methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, t-butanol, ethyl acetate and other lower alkanols, glycerine, acetone, Ethoxy ethanol dichloromethane, Dimethyl sulphoxide (DMSO), Dimethyl acetate (DMA), dimethyl formamide (DMF), isopropyl ether, methyl ethyl ketone, acetonitrile, toluene, N- methylpyrrolidone (NMP), tetrahydrofuran (THF), tetrahydropyran, water, other cyclic mono-, di- and tri-ethers, polyalkylene glycols '(e. g., polyethylene glycol, polypropylene glycol, propylene glycol) and mixtures thereof in suitable proportions. In an embodiment, the solvate of a taxane includes acetone or ethyl acetate. In an embodiment, the cabazitaxel solvate is an acetone solvate or an ethyl acetate solvate.

[0055] As used herein, the term “cancer” refers to one of a group of diseases caused by the uncontrolled, abnormal growth of cells that can spread to adjoining tissues or other parts of the body. Cancer cells can form a solid tumor, in which the cancer cells are massed together, or exist as dispersed cells. In the case of prostate cancer, the cells undergoing uncontrolled, abnormal growth originate in the prostate. The prostate cancer may be of any stage (early stage, locally advanced, or advanced), and may optionally be metastatic prostate cancer, relapsed prostate cancer, refractory prostate cancer, prostate cancer with acquired chemoresistance, and/or castrationresistant prostate cancer (CRPC). In an embodiment, the prostate cancer is advanced prostate cancer, metastatic prostate cancer, relapsed prostate cancer, refractory prostate cancer, prostate cancer with acquired chemoresistance, and/or CRPC. As used herein, “refractory prostate cancer” means a prostate cancer that is intrinsically refractive to treatment, i.e. the cancer is not sensitive to a particular chemotherapy upon first exposure to the chemotherapy. As used herein “relapsed prostate cancer” means a prostate cancerthat relapses and begins to grow again upon discontinuation of a particular chemotherapy. As used herein, “acquired chemoresistance” is an active process that develops within a cancer during treatment that makes the cancer less or entirely non responsive to a given chemotherapy. As used in the art, CRPC refers to prostate cancer that continues to grow even in the absence of normal levels of testosterone.

[0056] The term “cancer cell” refers a cell characterized by uncontrolled, abnormal growth and the ability to invade another tissue or a cell derived from such a cell. Cancer cells include, for example, a primary cancer cell obtained from a subject with cancer or cell line derived from such a cell.

[0057] The term "subject" as used herein includes all members of the animal kingdom including mammals, and suitably refers to humans. Optionally, the term “subject” includes mammals that have been diagnosed with prostate cancer or are in remission. In one embodiment, the term “subject” refers to a human having, or suspected of having, prostate cancer.

[0058] The term "subject in need thereof" refers to a subject that could benefit from the method(s) ortreatment(s) described herein, and optionally refers to a subject with prostate cancer, or optionally a subject with increased risk of prostate cancer, such as a subject previously having prostate cancer, a subject with a precancerous syndrome or a subject with a strong genetic disposition.

[0059] The term "increased risk of prostate cancer" as used herein means a subject that has a higher risk of developing prostate cancer than the average risk of the population. A subject may have a higher risk due to previously having had prostate cancer and or having a genetic risk factor for prostate cancer or exhibit a pre-cancer syndrome.

[0060] In one embodiment, the methods and uses described herein provide for the treatment of prostate cancer. The term "treating" or “treatment” as used herein and as is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease (e.g. maintaining a subject in remission), preventing disease or preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission (whether partial or total), whether detectable or undetectable. "Treating" and "treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment. "Treating" and "treatment" as used herein also includes prophylactic treatment. “Treating” and “treatment” as used herein can also mean reducing the cancer burden in a subject.

[0061] The phrase "cancer burden" refers to the quantum of cancer cells or cancer volume in a subject. Reducing cancer burden accordingly refers to reducing the number of cancer cells or the cancer volume in a subject.

[0062] The term "administered" or “administering” as used herein means administration of a therapeutically effective amount of a compound or composition of the disclosure to a cell either in cell culture or in a subject. [0063] The term “co-administration” or “combination therapy” means that at least two compounds or compositions are administered to the subject concurrently, such that effective amounts or concentrations of each of the two or more compounds may be found in the subject at a given point in time. Although compounds according to the present disclosure may be co-administered to a subject at the same time, the term embraces both administration of two or more agents at the same time or at different times, provided that effective concentrations of all co-administered compounds or compositions are found in the subject at a given time. “Coadministration” or “combination therapy” also embraces the use or administration of two or more agents in a single administration or a series of administrations, and/or in the same dosage form or separate dosage forms.

[0064] As used herein, the phrase "effective amount" or "therapeutically effective amount" means an amount effective, at dosages and frequencies, and for periods of time necessary to achieve the desired result. For example in the context of treating cancer, an effective amount is an amount that for example induces remission, reduces tumor burden, and/or prevents tumor spread or growth of cancer cells compared to the response obtained without administration of the compound. Effective amounts may vary according to factors such as the disease state, age, sex and weight of the animal. The amount of a given compound that will correspond to such an amount will vary depending upon various factors, such as the given drug or compound, the pharmaceutical formulation, the route of administration, the type of disease or disorder, the identity of the subject or host being treated, and the like, but can nevertheless be routinely determined by one skilled in the art.

[0065] Examples of suitable dosage ranges for cepharanthine free base may include for example up to 100mg. Cepharanthine free base is available in Japan in tablet form at 1 and 10mg. Cepharanthine free base has been safely used in healthy volunteers via oral and IV administration at up to 100mg per day for seven days without noted adverse events. An IV administration of 100 mg results in an average CMax of 6.9uM (Yasuda et al. 1989). Oral administration of enteric formulations may achieve therapeutic circulating concentration of for example about 6.9uM with doses in the 50- 400 mg/day range. Lower dosages may also be used. Accordingly, suitable dosages and dosage ranges include about 5 mg/day to about 400 mg/day, optionally about 10 mg/day, about 15 mg/day, about 20 mg/day, about 25 mg/day, about 50 mg/day, about 100 mg/day, about 150 mg/day, about 200 mg/day, about 300 mg/day, about 400 mg/day, or any suitable dose between about 5 mg/day to about 400 mg/day. Dosages of cepharanthine may be based on subject weight. Daily dosages of about 3 mg/kg body weight to about 27 mg/kg body weight are shown herein to be effective in a mouse model. Suitable dosages in humans can be estimated for example by calculating the human equivalent dose (HED) using for example the methods of Nair & Jacob (2016). Accordingly, suitable dosages of cepharanthine include, for example about 0.1 mg/kg body weight to about 4 mg/kg body weight, optionally about 0.244 mg/kg body weight to about 2.195 mg/kg body weight, or about 0.244 mg/kg body weight, about 0.732 mg/kg body weight, about 2.195 mg/kg body weight, or any suitable dosage between about 0.1 mg/kg body weight to about 4 mg/kg body weight. Examples of suitable dosage ranges for taxanes can be determined by the skilled person. For example, the recommended dose of cabazitaxel (JEVTANA) is 20 mg/m ² administered as a 1 -hour intravenous infusion every 3 weeks. A dose of 25 mg/m ² can be used in select subjects at the discretion of the treating healthcare provider. Paclitaxel is typically dosed at 135-175 mg/m ² at 3-week intervals. Docetaxel is typically dosed at 75-100 mg/m ² at 3-week intervals. Other dosages and intervals are specifically contemplated herein. Optionally, lower doses of cabazitaxel, paclitaxel, or docetaxel may be used in combination with cepharanthine and/or lower doses of cepharanthine may be used in combination with cabazitaxel, paclitaxel, or docetaxel.

[0066] Such dosages may be administered according to any suitable schedule for example daily, weekly, bi-weekly, monthly, or any other suitable frequency, depending on the given drug combination, formulation(s) and/or route(s) of administration, stage of disease, and/or subject-specific factors. Administration of cepharanthine may occur on the same schedule as or a different schedule from the taxane. For example, cepharanthine may be administered daily, and the taxane may be administered at 3-day intervals, weekly intervals, or 3-week intervals. Examples of suitable dosing schedules can be determined by the skilled person.

III. Compositions and Kits

[0067] As shown herein, the combination of cepharanthine and cabazitaxel act synergistically to inhibit the growth of prostate cancer cells. Accordingly, in one aspect there is provided a composition comprising cepharanthine, or a pharmaceutically acceptable salt or solvate thereof, a taxane, or pharmaceutically acceptable solvate thereof, optionally paclitaxel, docetaxel, or cabazitaxel, and a pharmaceutically acceptable carrier. Also provided are kits comprising cepharanthine, or pharmaceutically acceptable salt or solvate thereof, and a taxane, or pharmaceutically acceptable solvate thereof, optionally paclitaxel, docetaxel, or cabazitaxel, as described herein, optionally along with suitable container or packaging and/or instructions for the use thereof, such as for the treatment of prostate cancer in a subject in need thereof.

[0068] The cepharanthine, or pharmaceutically acceptable salt or solvate thereof, and taxane, or pharmaceutically acceptable solvate thereof, may be provided for administration in the same dosage form or alternatively may be provided as separate dosage forms. In an embodiment, the cepharanthine and taxane are provided as a composition. In an embodiment, the cepharanthine is provided as an oral dosage form.

[0069] The composition may be formulated for use or prepared for administration to a subject using pharmaceutically acceptable formulations known in the art. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003 - 20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999. The term “pharmaceutically acceptable” means compatible with the treatment of animals, in particular, humans.

[0070] The term “dosage form” as used herein refers to the physical form of a dose for example comprising a compound of the disclosure, and includes without limitation injectable dosage forms, including, for example, sterile solutions and sterile powders for reconstitution, and the like, that are suitably formulated for injection, liquid and solid dosage forms including, for example tablets, including enteric coated tablets, caplets, gelcaps, capsules, ingestible tablets, buccal tablets, troches, elixirs, suspensions, syrups, wafers, resuspendable powders, liquids and solutions.

[0071] On this basis, the pharmaceutical compositions could include an active compound or substance, such as a compound described herein, in association with one or more pharmaceutically acceptable vehicles or diluents, and/or contained in buffered solutions with a suitable pH and isoosmotic with the physiological fluids. The composition could include a targeting agent for the delivery or transport of the active compound to specified sites within the body, organ, tissue, or cell. [0072] As used herein, the term “diluent” refers to a pharmaceutically acceptable carrier which does not inhibit a physiological activity or property of an active compound to be administered and does not irritate the subject and does not abrogate the biological activity and properties of the administered compound. Diluents include any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservative salts, preservatives, binders, excipients, disintegration agents, lubricants, such like materials and combinations thereof, as would be known to one of ordinary skill in the art (see, for example, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329, incorporated herein by reference). Except insofar as any conventional carrier is incompatible with the active ingredient, its use in the pharmaceutical compositions is contemplated.

[0073] The pharmaceutical compositions, formulations, dosages, etc. described herein can be administered for example, by parenteral, intravenous, subcutaneous, intramuscular, intraorbital, ophthalmic, intraocular, intravitreal, intracameral, subtenon, subconjunctival, intraperitoneal, inhalation or spray (e.g. via aerosol), rectal or oral administration in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. The pharmaceutical compositions etc. described herein can be administered as a bolus dose, or as a continuous infusion.

[0074] The term “parenteral” as used herein includes percutaneous, subcutaneous, intravascular (e.g., intravenous), intramuscular, or intrathecal injection or infusion techniques and the like. In addition, there is provided a pharmaceutical formulation comprising an active compound, derivative, analog, pharmaceutically acceptable salt thereof or a mixture of any of the foregoing and a pharmaceutically acceptable carrier. One or more molecules of the disclosure can be present in association with one or more non-toxic pharmaceutically acceptable carriers and/or diluents and/or adjuvants, and if desired other active ingredients. The pharmaceutical compositions of the disclosure can be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs.

[0075] Compositions or dosage forms intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions can contain one or more such sweetening agents, flavoring agents, coloring agents or preservative agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients can be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets can be uncoated or they can be coated by known techniques. In some cases such coatings can be prepared by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monosterate or glyceryl distearate can be employed. Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil. In the case of cepharanthine, methods for preparing pharmaceutically acceptable salts suitable for oral formulations, and particularly enteric formulations, are disclosed for example in United States Patent No. 10,576,077. Suitably, the enteric formulation for oral administration may comprise about 25% w/w cepharanthine.2HCI in about 75% w/w Eudragit L100 (1/1 :methacrylic acid/methyl methacrylate enteric copolymer).

[0076] Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydropropyl-methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents can be a naturally-occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions can also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0077] Oily suspensions can be formulated by suspending the active ingredients in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions can contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents and flavoring agents can be added to provide palatable oral preparations. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

[0078] Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents or suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, can also be present. Pharmaceutical compositions of the disclosure can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil or a mineral oil or mixtures of these. Suitable emulsifying agents can be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally- occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions can also contain sweetening and flavoring agents.

[0079] Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol, glucose or sucrose. Such formulations can also contain a demulcent, a preservative and flavoring and coloring agents. The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents that have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parentally acceptable diluent or solvent, for example as a solution in 1 ,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

[0080] In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811 , herein incorporated by reference in its entirety.

[0081] Oral or parenteral compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the disclosure are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

[0082] The preceding section is provided by way of example only and is not intended to be limiting on the scope of the present disclosure and appended claims. Additional objects and advantages associated with the compositions and methods of the present disclosure will be appreciated by one of ordinary skill in the art in light of the instant claims, description, and examples. For example, the various aspects and embodiments of the disclosure may be utilized in numerous combinations, all of which are expressly contemplated by the present description. These additional advantages, objects and embodiments are expressly included within the scope of the present disclosure. The publications and other materials used herein to illuminate the background of the disclosure, and in particular cases, to provide additional details respecting the practice, are incorporated by reference, and for convenience are listed in the appended reference section.

IV. Examples

Example 1. Sensitivity of prostate cancer cells to cepharanthine.2HCI, cabazitaxel, and cisplatin monotherapy

[0083] Each cell line was treated with cepharanthine or a standard chemotherapy drug as reference control and culture medium containing 0.25% [v/v] DMSO as vehicle control.

[0084] Table 1 : cell lines and experimental conditions

General cell culture reagents and plastic.

[0085] FBS, (Cat# FND500, ExCell Bio. Store at -20°C)

[0086] 96-Well Flat Clear Bottom Black Polystyrene TC-Treated Microplates (Cat# 3603, Corning).

[0087] CellTiter-Glo® Luminescent Cell Viability Assay (Cat# G7572, Promega. Store at -20°C). Substrate is sufficient for 1 ,000 assays at 100 pL per assay in 96-well plates. Including:

1 x 100 mL CellTiter-Glo® Buffer

1 x vial CellTiter-Glo® Substrate (lyophilized)

Reagent Preparation [0088] CellTiter-Glo Buffer was thawed and equilibrated to room temperature prior to use.

[0089] The lyophilized CellTiter-Glo Substrate was equilibrated to room temperature prior to use.

[0090] The appropriate volume (100 mL) of CellTiter-Glo Buffer was transferred into the amber bottle containing CellTiter-Glo Substrate to reconstitute the lyophilized enzyme/substrate mixture. This forms the CellTiter-Glo Reagent.

[0091 ] The contents were mixed by gently vortexing, swirling or by inverting the contents to obtain a homogeneous solution. The CellTiter-Glo Substrate should go into solution easily in less than one minute.

[0092] Cepharanthine-2HCL was dissolved in water to make 10 mM stock solution and then aliquoted and stored in -20°C freezer.

Determination of the half maximal inhibition concentration IC50

[0093] Cells were harvested during the logarithmic growth period and counted using Count-star. Cell concentrations were adjusted to 4.44*10 ⁴ cells/mL with respective culture medium. A 90 pL volume of cell suspension was added to two 96- well plates (plates A and B) with the final cell density of 4x1 o ³ cells/well. (cell concentration adjusted according to the data base or density optimization assay.)

Next day: For the plates of TO reading:

[0094] A 10 pL volume of culture medium was added to each well of plate A for TO reading. The plate and its content was equilibrated at RT for approximately 30 min. A 50 pL volume of CellTiter-Glo reagent was added to each well. The contents were mixed for 5 min on an orbital shaker to induce cell lysis. The plates were allowed to incubate at RT for 20 min to stabilize luminescent signal.

[0095] Luminescence (TO) was recorded using EnVision Multi Label Reader.

For the plates of test reading:

[0096] A 10xsolution of each drug was prepared in media with 3.16-fold serial dilutions to achieve 9 dose levels (Top working concentration: 100 pM cepharanthine, 100 pM cisplatin (as reference control), 10 pM cabazitaxel). Cabazitaxel was examined as a monotherapy at concentrations that ranged from 10uM to 1nM. Cepharanthine was examined as a monotherapy at concentrations that ranged from 100 uM to 10 nM. Cisplatin was used as an experimental control at 100uM to 10nM. A 10 pL (10x) volume of drug solution was dispensed in individual wells (triplicate for each drug concentration) of plate B. Test plate B was incubated for 72 hours in the humidified incubator at 37°C with 5% CO2. The plate and its contents were equilibrated at RT for approximately 30 min. A 50 pL CellTiter-Glo reagent was added to each well. The contents were mixed for 5 min on an orbital shaker to induce cell lysis. The plates were allowed to incubate at RT for 20 min to stabilize luminescent signal. Luminescence (T3) was recorded using EnVision Multi Label Reader.

Data Analysis

[0097] The data was displayed graphically using GraphPad Prism 5.0.

[0098] In order to calculate absolute IC50 (EC50), a dose-response curve was fitted using nonlinear regression model with a sigmoidal dose response. The formula for calculating surviving rate is shown below and the absolute IC50 (EC50) was calculated according to the dose-response curve generated by GraphPad Prism 5.0.

Surviving rate (%) ^— (LumTest article-LumMedium control)/ (LumNon-treated-LUITlMedium control)* 100%.

[0099] Table 2. Summary of Absolute IC50s & Maximal inhibition in 3 cell lines

[00100] Dose-response curves for each cell line are shown in Figs. 1-3.

Example 2. Dose response curves and combination index for combinations of cepharanthine and cabazitaxel

[00101] Experimental protocols were as described in Example 1. To explore additive or synergistic effects, combinations of cepharanthine + cabazitaxel were tested on 22Rv1 at the following concentrations: 10uM/100nM, 3.16uM/31.6nM, 1.0uM/10nM, 316nM/3.16nM, 100nM/1 nM respectively. IC50 values for cepharanthine and cabazitaxel determined in Example 1 were used to generate dose response curves and calculate the combination index. [00102] The Combination Index (Cl) and confidence interval was determined based on Zhao's formulation ^{1 2} . Cl<1 indicates a synergistic effect, while Cl>1 indicates an antagonistic effect and Cl=1 means an additive effect. Cl and its 95% confidence interval are shown against % inhibition of the combination treatment, and can be used to identify optimal dosage range for drug synergy.

[00103] Table 3: Combination Index values generated according to Zhao’s formula based on extrapolation at the indicated inhibition value.

Inhibition CI lower upper

7.735 1.570 0.369 5.872

12.735 1.201 0.473 2.255

17.735 1.097 0.575 1.614

22.735 1.047 0.621 1.452

27.735 1.017 0.655 1.360

32.735 0.997 0.686 1.293

37.735 0.981 0.717 1.242

42.735 0.969 0.735 1.210

47.735 0.958 0.737 1.192

52.735 0.948 0.728 1.196

57.735 0.937 0.708 1.239

62.735 0.925 0.678 1.301

67.735 0.908 0.641 1.397

72.735 0.883 0.595 1.476

77.735 0.840 0.524 1.584

82.735 0.750 0.360 1.713

[00104] Table 4: IC50 and Cl values for Cepharanthine and Cabazitaxel in 22Rv1 prostate cancer cells.

Cell line drug A IC50A drug B IC50B CI_IC50 lower upper (pM) (pM)

22Rv1 Cepharanthin 4.396 Cabazitaxel 0.00956 0.95 0.73 1.19 e-2HCL

[00105] Growth inhibition curves for cepharanthine.2HCI, cabazitaxel, and cepharanthine.2HCI+cabazitaxel are shown in Figs. 4A-4C. The Combination Index (Cl) plot is shown in Fig. 4D. The curve-shift analysis plot is shown in Fig. 4E. For the curve-shift analysis, a left shift indicates a synergistic effect, while a right shift indicates an antagonistic effect, and no shift means an additive effect. As shown in Fig. 4E, the cepharanthine + cabazitaxel combination is shifted to the left compared to either drug alone, indicating a synergistic effect. Example 3. In Vivo Efficacy Study Of Cepharanthine in the Treatment of Subcutaneous 22Rv1 Human Prostate Cancer Xenograft Model In Male BALB/C Nude Mice

[00106] Experimental Design: Treatment plan of this study is shown as Table 5 below.

Table 5: Treatment plan for 22Rv1 model study. ROA = route of administration; i.p. = intraperitoneal; p.o. = oral; QD = once per day; Q3D = every three days (one day dosing and 2 days off). [00107] Experimental Methods

[00108] Cell Culture'. The 22Rv1 cells were maintained in vitro with RPMI1640 medium supplemented with 10% fetal bovine serum at 37°C in an atmosphere of 5% CO2 in air. The cells in exponential growth phase were harvested and quantitated by cell counter before tumor inoculation. [00109] Tumor Inoculation'. Each mouse was inoculated subcutaneously in the right upper flank region with 22Rv1 cells (1x 107) in 0.1 ml of PBS mixed with matrigel (1 : 1 ) for tumor development. [00110] Randomization: The randomization started when the mean tumor size reached approximately 121 mm ³ . In total, 80 mice were enrolled in the study and randomly allocated to 8 study groups as shown in Table 5, with 10 mice per group. Randomization was performed based on “Matched distribution” method (Study Director™ software, version 3.1.399.19). The date of randomization was denoted as day 0.

[00111] Observation and Data Collection: After tumor inoculation, the animals were checked daily for morbidity and mortality. During routine monitoring, the animals were checked for any effects of tumor growth and treatments on behavior such as mobility, food and water consumption, body weight gain/loss (Body weights were measured twice per week after randomization), eye/hair matting and any other abnormalities. Mortality and observed clinical signs were recorded for individual animals in detail.

[00112] Tumor volumes were measured twice per week after randomization in two dimensions using a caliper, and the volume was expressed in mm ³ using the formula: “V = (L x W x W)/2, where V is tumor volume, L is tumor length (the longest tumor dimension) and W is tumor width (the longest tumor dimension perpendicular to L). Dosing as well as tumor and body weight measurements were conducted in a Laminar Flow Cabinet.

[00113] The body weights and tumor volumes were measured by using StudyDirector™ software (version 3.1.399.19).

[00114] Drug Formulation

[00115] Dosing volume was adjusted for body weight (dosing volume = 10 pL/g)

Table 6: Drug formulations.

[00116] By way of example: Formulation methods for Cepharanthine (2.7 mg/mL) a) Accurately weigh about 48.79 mg (based on 24.9% payload) sample of CEPN.2HCI (enteric coated) into an appropriately sized glass bottle, add a magnetic stir bar. Add 4.5 mL sterile water for injection into the bottle, cap and vortex for 30 seconds. Alternatively, slowly add the water (approximately half of the 4.5 mL), then turn on the stir plate. Continue to gradually add water. Use a spatula to help mix all the CEPN.2HCI. Add the rest of the water to achieve a homogenous suspension. b) Magnetically stir the mixture slowly to avoid foaming of the test article. c) Use an appropriately sized syringe with an attached 16-gauge needle, draw a sample into the syringe. Tap and expel air bubbles. Stop at the desired volume. d) Use the suspension formulation within 35 min of preparation.

[00117] Article Administration'. The treatment was initiated on the same day of randomization (day 0) per Table 5.

[00118] Statistical Analysis’. To compare tumor volumes of different groups at a pre-specified day, Bartlett's test was used first to check the assumption of homogeneity of variance across all groups. When the p-value of Bartlett's test was >= 0.05, a one-way ANOVA was run to test overall equality of means across all groups. If the p-value of the one-way ANOVA was < 0.05, post hoc testing was performed by running Tukey's HSD (honest significant difference) tests for all pairwise comparisons, and Dunnett's tests were performed for comparing each treatment group with the vehicle group. When the p-value of Bartlett's test was <0.05, Kruskal-Wallis test was run to test overall equality of medians among all groups. If the p-value the Kruskal- Wallis test was <0.05, post hoc testing was performed by running Conover's nonparametric test for all pairwise comparisons or for comparing each treatment group with the vehicle group, both with single-step p-value adjustment. [00119] In addition, pairwise comparisons were performed without multiple comparison correction and nominal/uncorrected p-values were reported directly from Welch's t-test or Mann-Whitney U test. Specifically, Bartlett's test was used first to check the assumption of homogeneity of variance for a pair of groups. When the p- value of Bartlett's test was >0.05, Welch's t-test was run, otherwise Mann-Whitney U test was run, to obtain nominal p-values.

[00120] To compare body weight changes of different groups at different time points, two-way ANOVA Tukey’s multiple comparisons test was performed in Mixed- effects model with Geisser-Greenhouse correction to obtain adjusted p-values.

[00121] All statistical analyses were done in R-a language and environment for statistical computing and graphics (version 3.3.1 ). All tests were two-sided unless otherwise specified, and p-values of <0.05 are regarded as statistically significant.

[00122] For survival analysis, the survival time was analyzed by the Kaplan- Meier method. The survival time was defined as the time from the day of randomization until animal death or ethical endpoint. For each group, the median survival time (MST) and the increased in life-span (ILS) was calculated. The Kaplan-Meier curves were constructed for each group, the log-rank test and Gehan-Breslow-Wilcoxon test were used to compare survival curves between groups to obtain p-values, p-values of <0.05 are regarded as statistically significant.

[00123] Results

[00124] Tumor volume growth curves are shown in Fig. 5A - Fig. 5C.

[00125] Tumor Growth Inhibition: Tumor growth inhibition (TGI) and

Treatment/Control (T/C) calculations based on the tumor size data comparing to vehicle group are shown in Table 7 - Table 13.

[00126] Table 7. TGI and T/C calculation on Day 3.

Note: First, Bartlett’s test was performed to test homogeneity of variance and normality, P = 0.365 non-significant, therefore, the data followed a parametric test to compare groups. Second, ANOVA test was performed. P=>0.05 is considered not significant.

[00127] Table 8. TGI and T/C calculation on Day 7. Note: First, Bartlett’s test was performed to test homogeneity of variance and normality, P =0.316 non-significant, therefore, the data followed a parametric test to compare groups. Second, ANOVA test was performed. P=>0.05 is considered not significant.

[00128] Table 9. TGI and T/C calculation on Day 10.

Note: First, Bartlett’s test was performed to test homogeneity of variance and normality, P =0.315 non-significant, therefore, the data followed a parametric test to compare groups.

Second, ANOVA test was performed. P=<0.05 is considered statistically significant.

[00129] Table 10. TGI and T/C calculation on Day 14.

Note: First, Bartlett’s test was performed to test homogeneity of variance and normality, P =0.737 non-significant, therefore, the data followed a parametric test to compare groups. Second, ANOVA test was performed. P=<0.05 is considered statistically significant. [00130] Table 11. TGI and T/C calculation on Day 17.

Note: First, Bartlett’s test was performed to test homogeneity of variance and normality, P =0.816 non-significant, therefore, the data followed a parametric test to compare groups. Second, ANOVA test was performed. P=<0.05 is considered statistically significant.

[00131] Table 12. TGI and T/C calculation on Day 21. Note: First, Bartlett’s test was performed to test homogeneity of variance and normality, P =0.723 non-significant, therefore, the data followed a parametric test to compare groups. Second, ANOVA test was performed. P=<0.05 is considered statistically significant.

[00132] Table 13. TGI and T/C calculation on Day 28.

Note: First, Bartlett’s test was performed to test homogeneity of variance and normality, P =0.223 non-significant, therefore, the data followed a parametric test to compare groups. Second, ANOVA test was performed. P=>0.05 is considered not statistically significant.

[00133] Body weight change percentage curves are shown in Fig. 6A - Fig. 6B. [00134] Table 14. Comparison of body weight between group 5 to 8 using Two

Way ANOVA Tukey's multiple comparisons test.

[00135] Mortality and tolerability: Treatments of Cabazitaxel or Cabazitaxel combined with Cepharanthine (group 5 to 8) induced large body weight loss (BWL) in mice. Mortality related to treatment was observed in group 5, 6, 7 and 8 since day 17 (group 8, two mice euthanized due to BWL>20%) resulting in 1/10, 1/10, 1/10 and 3/10 deaths respectively on day 21 ; while during additional 7 days’ observation period after 21 days’ treatment completion higher mortality rate was observed resulting in 4/10, 6/10, 8/10 and 6/10 deaths in group 5 to 8 respectively. No significant difference was found when comparing body weight of each other group between group 5 to 8. Control and Cepharanthine only treatments (groups 1 to 4) were well-tolerated.

[00136] Table 15. Test article related mortality and tolerability. [00137] Survival analysis: No significant difference was observed in median survival time of mice among group 5, 6, 7 and 8.

[00138] Table 16. Statistical analysis of survival between different treatment groups.

[00139] Summary: The objective of this study was to evaluate preclinically and confirm the in vivo therapeutic efficacy of Cepharanthine single, Cabazitaxel single and Cepharanthine combined cabazitaxel administrations in the treatment of the subcutaneous 22Rv1 human prostate cancer xenograft model in male BALB/c nude mice.

[00140] Cabazitaxel combined Cepharanthine groups (group 6 to 8) demonstrated significant anti-tumor effects compared to the control group using data of day 21 , the TGIs were 57.86%, 63.86% and 56.56%. Cabazitaxel single and Cepharanthine single administrations did not demonstrate significant anti-tumor effect at the time points examined (p>0.05). No significant difference was observed in median survival time of mice among group 5, 6, 7 and 8. Treatments of Cabazitaxel or Cabazitaxel combined Cepharanthine (group 5 to 8) induced large body weight loss (BWL) in mice resulting in mortalities. Mice from groups 1 to 4 tolerated treatments well. The addition of cepharanthine alone (groups 2-4 vs group 1 ), or in combination with cabazitaxel (groups 6-8 vs group 5), did not statistically worsen weight loss compared to the relative control group.

[00141] While the present application has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole. [00142] All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.

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