WO2022032146A1

WO2022032146A1 - Methods for inhibiting growth of era positive cancers

Info

Publication number: WO2022032146A1
Application number: PCT/US2021/045023
Authority: WO
Inventors: Ramon Moreno; Elizabeth Bruckheimer; Theodore M. Tarasow
Original assignee: Systems Oncology, Llc
Priority date: 2020-08-07
Filing date: 2021-08-06
Publication date: 2022-02-10

Abstract

Methods of treating and inhibiting growth of ERa positive tumors and cancers are described herein.

Description

METHODS FOR INHIBITING GROWTH OF ERa POSITIVE CANCERS

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to and benefit of U.S. Provisional Patent Application No. 63/062,856, filed August 7, 2020, the disclosures of which are hereby incorporated herein by reference in their entirety.

FIELD OF THE INVENTION

The present invention relates to methods of treating and inhibiting growth of ERa positive tumors and cancers.

BACKGROUND

ERa positive cancers, which include breast, ovarian, uterine, cervical carcinoma, and endometrial cancers, are difficult to treat, with high rates of recurrence and drug resistance. ERa positive cancer cells can remain quiescent for extended periods of time and then reactivate. Endocrine therapies such as fulvestrant and tamoxifen that have been used to treat ERa positive cancers are cytostatic drugs, not cytotoxic drugs, and do not kill residual dormant ERa positive cancer cells, which can later proliferate causing recurrence. Cytotoxic anticancer drug therapies are used to kill tumor cells, including residual tumor cells. However, the toxicities of cytotoxic anticancer drugs that make them effective at killing cancer cells, also have frequent and severe side effects at therapeutic doses, causing toxicities such as haematological, gastrointestinal, skin and hair follicle toxicity, nervous system toxicity, local toxicity, metabolic abnormalities, hepatic toxicity, urinary tract toxicity, cardiac toxicity, pulmonary toxicity, and gonadal toxicity. Remesh, A., Int J Basic Clin Pharmacol;. 2012 1 (1):2- 12; Taskln-Tok, T. et al., “Anticancer Drug - Friend or Fee”, Pharmacology and Therapeutics, 2014, Chapter 9, pp. 255-269. The need to limit anticancer drug toxicities can lead to a reduction in the efficacy of the anticancer drug.

There is a need for methods of inhibiting growth of ERa positive tumors and cancers using cytotoxic small molecule drugs where drug efficacy is maximized while minimizing dose limiting toxicity.

SUMMARY OF THE INVENTION

The present invention relates to a method of ameliorating or treating an ERa positive cancer in a human by administering to the human patient a compound having the formula

(hereinafter designated “ErSO compound”) once a week in a therapeutically effective amount greater than 1 .5 mg/kg of body weight when administered orally or in a therapeutically effective amount of at least 0.4 mg/kg of body weight when administered intravenously, with the weekly dose repeated until there is complete regression of the ERa positive cancer.

In some embodiments, the ErSO compound is administered orally to a human patient in a therapeutically effective amount of 1 .6 to 4.9 mg/kg of body weight, 1 .6 to 4.1 mg/kg of body weight, of 1 .6 to 3.3 mg/kg of body weight, 2.0 to 3.3 mg/kg of body weight, 2.4 to 3.3 mg/kg of body weight, or 3.3 mg/kg of body weight.

In some embodiments, the ErSO compound is administered intravenously to a human patient in a therapeutically effective amount of 0.4 to less than 1 .6 mg/kg of body weight or in a therapeutically effective amount of 0.4 to 0.8 mg/kg of body weight.

The present invention relates to the use of an ErSO compound in a method of ameliorating or treating an ERa positive cancer in a human patient in which the compound is administered to the human patient once a week in a therapeutically effective amount greater than 1 .5 mg/kg of body weight when administered orally or a therapeutically effective amount of at least 0.4 mg/kg of body weight when administered intravenously, with the weekly dose is repeated until there is complete regression or until growth of the ERa positive cancer is inhibited.

The present invention relates to a method of inhibiting an ERa positive tumor in a mammal by administering to the mammal an ErSO compound once a week in an amount greater than 20 mg/kg of body weight when administered orally or in an amount of at least 5 mg/kg of body weight when administered intravenously, with the weekly dose repeated until growth of the ERa positive tumor is inhibited.

In some embodiments, the ErSO compound is administered orally to a mammal in a therapeutically effective amount of 20 to 60 mg/kg of body weight, 20 to 50 mg/kg of body weight, of 20 to 40 mg/kg of body weight, 25 to 40 mg/kg of body weight, 30 to 40 mg/kg of body weight, or 40 mg/kg of body weight.

In some embodiments, the ErSO compound is administered intravenously in a therapeutically effective amount of 5 to less than 20 mg/kg of body weight or 5 to 10 mg/kg of body weight. In some embodiments, the weekly dose of the ErSO compound is repeated until or until growth of the ERa positive cancer is inhibited.

In some embodiments, the ERa positive cancer is a breast cancer, an ovarian cancer, a uterine cancer, a cervical carcinoma, or an endometrial cancer.

BRIEF DESCRIPTION OF THE DRAWINGS The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

Figure 1 is a graph of tumor doubling time of tumor type MCF-7 with oral (PO) administration of control ErSO vehicle (Log™ Mean Tumor Volume) over 22.4 days (Td ~ 22.4 days). R² = 0.9285.

Figure 2 is a graph of animal weights (mean ± SD) for animals that received oral (PO) administration daily for 21 days of either ErSO vehicle, fulvestrant, 40 mg/kg ErSO, 30 mg/kg ErSO, 20 mg/kg ErSO, or 10 mg/kg ErSO.

Figure 3 is a graph of animal weights (mean ± SD) for animals that received oral (PO) administration weekly three times of either ErSO vehicle, fulvestrant, 40 mg/kg ErSO, 30 mg/kg ErSO, 20 mg/kg ErSO, or 10 mg/kg ErSO.

Figure 4 is a graph of animal weights (mean ± SD) for animals that received intravenous (IV) administration daily for 21 days of either ErSO vehicle, fulvestrant, 10 mg/kg ErSO, 5 mg/kg ErSO, 1 mg/kg ErSO, or 0.5 mg/kg ErSO. Figure 5 is a graph of animal weights (mean + SD) for animals that received intravenous (IV) administration weekly three times of either ErSO vehicle, fulvestrant, 10 mg/kg ErSO, 5 mg/kg ErSO, 1 mg/kg ErSO, or 0.5 mg/kg ErSO.

Figure 6 is a graph of percentage animal weight change vs. Day 0 for animals that received oral (PO) administration daily for 21 days of either ErSO vehicle, fulvestrant, 40 mg/kg ErSO, 30 mg/kg ErSO, 20 mg/kg ErSO, or 10 mg/kg ErSO. Figure 7 is a graph of percentage anima! weight change vs. Day 0 for animals that received oral (PO) administration weekly three times of either ErSO vehicle, fulvestrant, 40 mg/kg ErSO, 30 mg/kg ErSO, 20 mg/kg ErSO, or 10 mg/kg ErSO.

Figure 8 is a graph of percentage animal weight change vs. Day 0 for animals that received intravenous (IV) administration daily for 21 days of either ErSO vehicle, fulvestrant, 10 mg/kg ErSO, 5 mg/kg ErSO, 1 mg/kg ErSO, or 0.5 mg/kg ErSO.

Figure 9 is a graph of percentage animal weight change vs. Day 0 for animals that received intravenous (IV) administration weekly three times of either ErSO vehicle, fulvestrant, 10 mg/kg ErSO, 5 mg/kg ErSO, 1 mg/kg ErSO, or 0.5 mg/kg ErSO.

Figure 10 is a graph of tumor volume (Mean ± SEM) for animals that received oral (PO) administration daily for 21 days of either ErSO vehicle, fulvestrant, 40 mg/kg ErSO, 30 mg/kg ErSO, 20 mg/kg ErSO, or 10 mg/kg ErSO.

Figure 11 is a graph of tumor volume (Mean ± SEMI) for animals that received oral (PO) administration weekly three times of either ErSO vehicle, fulvestrant, 40 mg/kg ErSO, 30 mg/kg ErSO, 20 mg/kg ErSO, or 10 mg/kg ErSO.

Figure 12 is a graph of tumor volume (Mean ± SEM) for animals that received intravenous (IV) administration daily for 21 days of either ErSO vehicle, fulvestrant, 10 mg/kg ErSO, 5 mg/kg ErSO, 1 mg/kg ErSO, or 0.5 mg/kg ErSO.

Figure 13 is a graph of tumor volume (Mean ± SEM) for animals that received intravenous (IV) administration weekly three times of either ErSO vehicle, fulvestrant, 10 mg/kg ErSO, 5 mg/kg ErSO, 1 mg/kg ErSO, or 0.5 mg/kg ErSO.

Figures 14A-C are graphs. Figure 14A is a graph of the percent (%) dissolution of 30 mg of ErSO for seven prototypes over 24 hours (Time (hrs)) in simulated gastric fluid (SGF). The seven prototypes are designated A5, A6, A7, A8, A9, A10 and API (ErSO API), each of which contained a dose of 30 mg ErSO. The compositions of these prototypes is provided in Table 5. Figure 14B is a graph of the percent (%) dissolution of 30 mg of ErSO for five prototypes over 24 hours (Time (hrs)) in fasted state small intestinal conditions fluid (FaSSIF). The five prototypes are designated A5, A7, A8, A10 and API (ErSO API), each of which contained a dose of 30 mg ErSO. The compositions of these prototypes is provided in Table 5. Figure 14C is a graph of the concentration of ErSO (cone ng/ml) over 24 hours (time (h)) for four different prototypes designated micronized (micronized ErSO API), A7, A8 and A10, each of which contained a dose of 30 mg ErSO. The compositions of these prototypes is provided in Table 5.

Figures 15A-D are graphs showing the estimated effective doses based on pharmacokinetic data from oral administration of four prototype formulations containing 30 mg doses of ErSO designated Micronized API (ErSO API) (Figure 15A), A7 LBF (A7 lipid based formulation (Figure 15B), A8 LBF (A8 lipid based formulation (Figure 15B), and A10 LBF (107 lipid based formulation (Figure 15D). The compositions of these prototypes is provided in Table 5. The three graphs in each of Figures 15A-D show from left to right: left graph shows the plasma concentration of ErSO from the specified prototype for two female beagle dogs identified as 8275406 and 8280951 from blood samples taken at 0.25 hr, 0.5 hr, 1-4 hr, 8-12 hr, and 24 hr; middle graph plots the concentration (ng/ml) of 30 mg dose of ErSO over a simulation time from 0 to 24 hours; right graph plots the concentration (ng/ml) of the effective mg dose of ErSO over a simulation time from 0 to 24 hours.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Compounds having anticancer activity against ERa positive cancer celis have been described in published PCT Application No. WO 2020/009958, which is incorporated by reference in its entirety. The ERa positive cancer can be a breast cancer, an ovarian cancer, a uterine cancer, a cervical carcinoma, or an endometrial cancer. These compounds were administered daily, with daily dosing being in a single dose or a subdivided single dose given in spaced administration throughout the day, typically for twenty-one days, included in these compounds to be administered daily is a compound having the formula:

hereinafter designated “ErSO compound”. it has been surprisingly found that administering the ErSO compound once a week administered orally or intravenously instead of daily administration of the ErSO compound results in inhibition of ERa positive cancer cells and no re-growth, thereby maximizing drug efficacy while minimizing dose limiting toxicity. The benefits of once weekly dosing include reduction in overall dosing of the compound, lower potential of dose limiting toxicity, improved exposure per dose, greater patient compliance, and fewer patient visits. The weekly dose of the ErSO compound can be repeated until there is complete regression of the ERa positive cancer.

When tested in mice, the ErSO compound can be administered orally in an amount of 20 to 60 mg/kg of body weight, 20 to 50 mg/kg of body weight, 20 to 40 mg/kg of body weight, 25 to 40 mg/kg of body weight, 30 to 40 mg/kg of body weight or 40 mg/kg of body weight. The ErSO compound can be administered intravenously in mice in an amount of 5 to less than 20 mg/kg of body weight, preferably 5 to 10 mg/kg of body weight. Techniques for scaling doses for size differences or converting doses between species are well known. See, e.g., U.S. Dept, of Health and Human Services, Food and Drug Administration (July 20025) “Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers”; Sharma, V. et al., “To scale or not to scale: the principles of dose extrapolation”, British J. Pharmacol. 2009, 157:9070921 ; Nair, A. et al., “A simple practice guide for dose conversion between animals and human”, J. Basic Clin. Pharm. 2016, 7(2): 27-31 ; Freireich, E.J. et al., Cancer Chemother Rep.1966, 50(4):219-244.

Using the FDA guidance to convert animal doses to human equivalent does based on body surface area, and assuming the reference human body weight is 60 kg with a body surface area of 1 .62 m², the mouse doses are divided by 12.3 to obtain the human equivalent does in Table 1 below.

Table 1

Based on these human equivalent doses, the ErSO compound can be administered once a week in humans in an amount greater than 1.5 mg/kg of body weight when administered orally, or in an amount of at least 0.4 mg/kg of body weight when administered intravenously instead of dally administration of these amounts of ErSO compound, with resulting inhibition of ERa positive cancer cells and no re-growth, thereby maximizing drug efficacy while minimizing dose limiting toxicity. The ErSO compound can be administered to humans orally in an amount of 1 .6 to 4.9. mg/kg of body weight, 1 .6 to 4.1 mg/kg of body weight, 1 .6 to 3.3 mg/kg of body weight, 2.0 to 3.3 mg/kg of body weight, 2.4 to 3.3 mg/kg of body weight or 3.3 mg/kg of body weight. The ErSO compound can be administered to humans intravenously in an amount of 0.4 to less than 1 .6 mg/kg of body weight, preferably 0.4 to 0.8 mg/kg of body weight. Selection of a therapeutically effective dose will be determined by the skilled artisan considering several factors, which will be known to one of ordinary skill in the art. Such factors include the particular form of the pharmacological agent, and its pharmacokinetic parameters such as bioavailability, metabolism, and half-life, which will have been established during the usual development procedures typically employed in obtaining regulatory approval for a pharmaceutical compound. Further factors in considering the dose include the condition or disease to be treated or the benefit to be achieved in a normal individual, the body mass of the patient, the route of administration, whether the administration is acute or chronic, concomitant medications, and other factors well known to affect the efficacy of administered pharmaceutical agents. Thus, the precise dose should be decided according to the judgment of the person of skill in the art, and each patient’s circumstances, and according to standard clinical techniques.

As used herein, the terms “therapeutically effective amount’’, “therapeutically effective dose” and “effective amount” refer to an amount of ErSO compound and compositions which is sufficient to effect beneficial or desired results, that, when administered alone or in combination with an additional therapeutic agent to a cell, tissue, or subject, is effective to cause a measurable improvement in one or more symptoms of an ERa positive cancer or in delaying, reducing or mitigating the progression of such ERa positive cancer. A therapeutically effective dose further refers to that amount of the compound sufficient to result in at least partial amelioration of symptoms, e.g., treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions. When applied to an individual active ingredient administered alone, a therapeutically effective dose refers to that ingredient alone. When applied to a combination, a therapeutically effective dose refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. The effective amount of the ErSO compound may be less when administered in a combination than when administered alone. An effective amount can also result in an improvement in a subjective measure in cases where subjective measures are used to assess disease severity.

The ErSO compound can be in the form of a pharmaceutical composition, which may comprise a therapeutically effective amount of the ErSO compound and a pharmaceutically acceptable carrier. Preferred methods of administration of the ErSO compound and compositions for use in the methods of the invention are oral, intrathecal, intratumoral and parental including intravenous. Further methods of administration may include subcutaneous, intra-muscular, intraperitoneal, or intravesicular administration to a subject. The ErSO compound must be in the appropriate form for administration of choice. The phrase "pharmaceutically acceptable" or “pharmacologically acceptable” as used herein refers to molecular entities and compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a human, and approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

The term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as saline solutions in water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. A saline solution is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The use of solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like for pharmaceutical active substances that are pharmaceutically acceptable as the term is used herein are well known in the art and are preferably inert. Except insofar as any conventional media or agent is incompatible with the active ingredients, its use in therapeutic compositions is contemplated.

Oral lipid-based drug delivery systems in which a drug is encapsulated or solubilized in lipid excipients can be used to increase solubilization and absorption of a drug, such as a poorly water soluble drug, to obtain enhanced bioavailability. Various lipid excipients and formulation approaches are described in Kalepu, S. et al., Acta Pharmaceutics Sinica B 2013, 3 (6): 361 -372.

Pharmaceutical compositions adapted for oral administration may be capsules, tablets, powders, granules, solutions, syrups, suspensions (in non-aqueous or aqueous liquids), or emulsions. Tablets or hard gelatin capsules may comprise lactose, starch or derivatives thereof, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, stearic acid or salts thereof. Soft gelatin capsules may comprise vegetable oils, waxes, fats, semisolid, or liquid polyols. Solutions and syrups may comprise water, polyols, and sugars. An active agent intended for oral administration may be coated with or admixed with a material that delays disintegration and/or absorption of the active agent in the gastrointestinal tract. Thus, the sustained release may be achieved over many hours and if necessary, the active agent can be protected from degradation within the stomach. Pharmaceutical compositions for oral administration may be formulated to facilitate release of an active agent at a particular gastrointestinal location due to specific pH or enzymatic conditions,

A further preferred form of administration is parenteral including intravenous administration. Pharmaceutical compositions adapted for parenteral administration, including intravenous administration, include aqueous and non-aqueous sterile injectable solutions or suspensions, which may contain anti-oxidants, butters, bacteriostats, and solutes that render the compositions substantially isotonic with the blood of the subject. Other components which may be present in such compositions include water, alcohols, polyols, glycerine, and vegetable oils. Compositions adapted for parental administration may be presented in unit- dose or multi-dose containers, such as sealed ampules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile carrier, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets. Suitable vehicles that can be used to provide parenteral dosage forms of the invention are well known to those skilled in the art. Examples include: Water for Injection USP; aqueous vehicles such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles such as ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate. Definitions

The terms used in this specification generally have their ordinary meanings in the art, within the context of this invention and the specific context, where each term is used. Certain terms are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner in describing the methods of the invention and how to use them. Moreover, it will be appreciated that the same thing can be said in more than one way. Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein, nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of the other synonyms. The use of examples anywhere in the specification, including examples of any terms discussed herein, is illustrative only, and in no way limits the scope and meaning of the invention or any exemplified term. Likewise, the invention is not limited to its preferred embodiments.

The term “subject” as used in this application means an animal with an immune system such as avians and mammals. Mammals include canines, felines, rodents, bovine, equines, porcines, ovines, and primates. Avians include, but are not limited to, fowls, songbirds, and raptors. Thus, the invention can be used in veterinary medicine, e.g., to treat companion animate, farm animals, laboratory animals in zoological parks, and animate in the wild. The invention is particularly desirable for human medical applications.

The term “patient” as used in this application means a human subject. In some embodiments of the present invention, the “patient” is suffering with cancer.

The term “in need thereof’ would be a subject known or suspected of having or being at risk of cancer.

The terms “treat”, “treating” or “treatment” of a state, disorder or condition includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder, or condition developing in a person who may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical symptoms of the state, disorder or condition; or (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical symptom, sign, or test, thereof; or (3) relieving the disease,

causing regression of the state, disorder or condition or at least one of its clinical or sub-clinical symptoms or signs.

The term “agent” as used herein means a substance that produces or is capable of producing an effect and would include, but is not limited to, chemicals, pharmaceuticals, biologies, small organic molecules, antibodies, nucleic acids, peptides, and proteins. The terms “agent”, “compound” and “drug” are interchangeable.

The term “an effective amount” refers to the amount of an agent, composition or drug that is sufficient to effect beneficial or desired results.

The term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system, i.e., the degree of precision required for a particular purpose, such as a pharmaceutical formulation. For example, “about” can mean within 1 or more than 1 standard deviations, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and more preferably still up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated, the term “about” meaning within an acceptable error range for the particular value should be assumed.

EXAMPLES

The present invention may be better understood by reference to the following nonlimiting examples, which are presented in order to more fully illustrate the preferred embodiments of the invention. They should in no way be construed to limit the broad scope of the invention.

EXAMPLE 1

Study Design

Antitumor activity of the ErSO compound and regimens were evaluated in a Cell- Derived Xenograft (CDX) model representing human breast cancer in immune-deficient female mice. The immune-deficient female mice were athymic nude, outbred homozygous (J:NU(Foxn1 ^nu/Foxn1 ”^u) (The Jackson Laboratory, Bar Harbor, ME). Data collected from the study included animal weights, observations, and tumor dimensions. The data was used to determine agent tolerability based on weight change and gross physiologic changes, as well as anticancer activity based on tumor growth inhibition or regression. The data analysis endpoint for this study was Day 45.

MCF-7 cells were cultured and approximately 20 x 10⁶ MCF-7 cells were injected into immune-deficient mice, with the study initiated at a mean tumor volume of approximately 125- 250 mm³. No tumor burden was associated with this model based on lack of weight loss or limited weight gain versus Day 0 in control group animals.

The ErSO compound was compared to the compound fulvestrant, sold under the brand name FASLODEX® (AstraZeneca, Wilmington, DE), and to the control vehicle (ErSO vehicle). The administration volume of the ErSO compound and control vehicle was 10 mL/kg. The vehicle for administration of the ErSO compound and the control was 5% DMSO, 10% Tween- 20, 85% PBS. Fulvestrant was administered in a dose of 2.5 mg/kg in a fixed volume of 2.5 mg/mL. Table 2 provides the detailed study outline with identifying group number for each group of animals tested (Group), the number of animals tested in each Group (-N-), the test agents (Tx), the doses given (Dose (mg/kg)), the route of administration (ROA) and schedule of doses (Schedule), the doses administered including the total number of doses (Total Dosed) and the day(s) of administration (Tx Day(s)), and the endpoint day (Endpoint). The route of administration for ErSO and the ErSO vehicle was either oral injection (PO) or intravenous (IV), and the route of administration for fulvestrant was subcutaneous (SC). The schedule of doses was either once a day for 21 days (qdx21) or once a week for three weeks (q7dx3).

Table 2

‘Dosed at a fixed-volume dose.

Weight and Agent Tolerability Analysis

For each group of animals (Group), Table 3 provides /he weight data on day 45, the weight nadir on day 45, and drug deaths. The weight data is presented as a mean for each group plus or minus standard deviation (Mean ± SD) and the percent weight change versus the Day 0 study initiation measurement (%vD₀). The weight nadir is presented as the maximum mean %vDo (%vDomax), which is the maximum weight loss versus the Day 0 study initiation measurement, on the day indicated in Table 3. The drug deaths are broken down into four types: (1) drug related animal death as a result of toxicity (D); (2) technical related animal death as a result of technician error (T); (3) tumor (burden) related animal death as a result of tumor-related weight loss or cachexia (B); and (4) unknown related animal death where cause of death cannot be determined (U).

Table 3

The data in Table 3 shows that ErSO administered alone by intravenous (IV) and oral (PO) injection and fulvestrant alone were well tolerated with slight weight loss and no drug- related deaths; deaths of unknown cause occurred in four groups at various times during the study. Figures 2, 3, 4 and 5 show that ErSO administered alone by intravenous (IV) and oral (PO) injection and fulvestrant alone, whether given daily for twenty-one days or weekly three times, did not result in significant weight loss. Figures 6, 7, 8 and 9 show that the percent weight change versus Day 0 was low when ErSO was administered alone by intravenous (IV) and oral (PO) injection and fulvestrant alone, whether given daily for twenty-one days or weekly three times.

Efficacy and Tumor Volume Analysis

Table 4 provides the data on efficacy of the compounds being tested and the data on tumor volume for each group of animals (Group). Tumor volume is presented as a mean for each group plus or minus standard error mean (Mean ± SEM). Tumor growth inhibition (%TGI) is the percent mean tumor growth versus Day 0 between treatment (TX) and control (C) groups using the formula

Table 3 also provides p-values, significance, partial responders (PR) and their percent tumor regression (%TR), complete responders (CR) and tumor free survivors (TFS). Partial responders had tumor volume >50% regression versus Day 0 for two consecutive measurements over a period of >7 days during or at completion of the study. The percent tumor regression was the percentage reduction in

Day 0 tumor volume calculated at the study endpoint, with a mean value used if multiple regressions. Complete responders had tumor volume <15 mm³ for two consecutive measurements over a period of >7 days during or at completion of the study. Tumor free survivors had durable complete response at completion of the study.

1 Versus ErSO vehicle (PO); ²versus ErSO vehicle (IV)

The data in Table 4 shows that ErSO dosed IV daily at 5 or 10 mg/kg or PO at 30 or 40 mg/kg for twenty-one days or once weekly for three doses resulted in similar, statistically significant (p<0.05) tumor growth inhibition. Fuivestrant was not active towards the MCF-7 model.

Animals dosed daily for twenty-one days included one partial tumor regression in Group 4 dosed PO at 40 mg/kg; two partial tumor regressions in Group 14 dosed IV at 5 mg/kg; three partial tumor regressions in Group 8 dosed PO at 20 mg/kg; five partial tumor regressions in Group 12 dosed IV at 10 mg/kg; two complete tumor regressions in Group 8 dosed PO at 20 mg/kg and Group 14 dosed IV at 5 mg/kg; four complete tumor regressions in Group 4 dosed PO at 40 mg/kg and Group 6 dosed PO at 30 mg/kg; and five complete tumor regressions in Group 12 dosed IV at 10 mg/kg.

Animals dosed once weekly for three doses included three partial tumor regressions in Group 7 dosed PO at 30 mg/kg, Group 9 dosed PO at 20 mg/kg and Group 15 dosed IV at 5 mg/kg; five partial tumor regressions in Group 13 dosed IV at 10 mg/kg; one complete tumor regression in Group 7 dosed PO at 30 mg/kg, Group 9 dosed PO at 20 mg/kg and Group 15 dosed IV at 5 mg/kg; two complete tumor regressions in Group 14 dosed PO at 20 mg/kg; four complete tumor regressions in Group 5 dosed PO at 40 mg/kg; and five complete tumor regressions in Group 13 dosed IV at 10 mg/kg.

Daily oral administration (PO) of ErSO for 21 days (qdx21) resulted in 100% complete regression at 40mg/kg (5/5) and 30mg/kg (4/5, with one mouse found dead on day 21), as well as 60% complete regression at 20mg/kg (3/5). Partial regression (40%) was observed in the 20mg/kg group (2/5). Figure 10 shows tumor volume (Mean ± SEM) for daily oral (PO) administration for 21 days of either ErSO vehicle, fuivestrant, ErSO 40 mg/kg, ErSO 30 mg/kg, ErSO 20 mg/kg, or ErSO 10 mg/kg.

Oral administration of ErSO once a week for 3 weeks (q7dx3) resulted in 100% complete regression at 40mg/kg (5/5) and 20% complete regression at 30mg/kg (1/5, with one mouse found dead on day 17) and 20mg/kg (1/5). Figure 1 1 shows tumor volume (Mean ± SEM) for weekly oral (PO) administration for three weeks of either ErSO vehicle, fuivestrant, ErSO 40 mg/kg, ErSO 30 mg/kg, ErSO 20 mg/kg, or ErSO 10 mg/kg.

Daily intravenous administration (IV) of ErSO for 21 days (qdx21) resulted in 100% complete regression at 10mg/kg (5/5), and 40% complete regression at 5mg/kg (2/5) at 5mg/kg. Partial regression (40%) was also observed at 5mg/kg (2/5). Figure 12 shows tumor volume (Mean ± SEM) for daily intravenous (IV) administration for 21 days of either ErSO vehicle, fuivestrant, ErSO 10 mg/kg, ErSO 5 mg/kg, ErSO 1 mg/kg, or ErSO 0.5 mg/kg.

Intravenous administration of ErSO once a week for 3 weeks (q7dx3) resulted in 100% complete regression at 10mg/kg (5/5), and 20% complete regression at 5mg/kg (1/5). Partial regression (60%) was observed at 5 mg/kg (3/5, mouse found dead on day 31). Figure 13 shows tumor volume (Mean ± SEM) for weekly intravenous (IV) administration given three times of either ErSO vehicie, fulvestrant, ErSO 10 mg/kg, ErSO 5 mg/kg, ErSO 1 mg/kg, or ErSO 0.5 mg/kg.

Overall, ErSO dosed once weekly for three doses at 5 or 10 mg/kg IV or 30 or 40 mg/kg PO was sufficient to produce significant tumor growth inhibition and tumor regressions.

EXAMPLE 2

Several lipid-based formulation samples were prepared to test for improved oral bioavailability for ErSO, the active pharmaceutical ingredient (ErSO API). ErSO API was a yellow solid characterized as a minimally hygroscopic, amorphous compound. ErSO has a molecular weight of 453.3 g/mol, a calculated pKa of 9.48, 1 1 .84, and a partition coefficient of 6,44. All active formulation samples were prepared by adding micronized ErSO API into a clear scintillation vial followed by the corresponding excipients under yellow light. All samples were then homogenized using a handheld homogenizer until the molecule was visually dissolved, for about 15 minutes. All samples were de-aerated to remove all the air bubbles and blanketed with nitrogen.

A total of six prototypes were prepared and evaluated. The compositions of these prototypes are shown in Table 5, Materials used in the compositions of the prototypes included (1) olive oil; (2) caprylocaproyl polyoxyl-8 glycerides NF sold under the brand name Labrasol® CC (Gattefosse Corp. Paramus, NJ); (3) lauroyl polyoxyl-32 glycerides sold under the name Gelucire® 44/14 CC (Gattefosse Corp. Paramus, NJ); (4) polysorbate 80 sold under the brand name Tween® 80 (Millipore Sigma, St. Louis, MO); (5) Vitamin E TPGS (tocopherosolan); propylene glycol monolaurate sold under the brand name Lauroglycol™ FCC (Gattefosse Corp. Paramus, NJ); (6) polyethylene glycol 400 (PEG 400); (7) glycerol/glyceryl monolinoleate sold under the brand name Maisine® CC (Gattefosse Corp. Paramus, NJ); (8) polyoxyl castor oil sold under the brand name Koiliphor® EL (BASF Corp. Floram Park, NJ); (9) polyoxyl 40 hydrogenated castor oil sold under the brand name Koiliphor® RH40 (BASF Corp. Floram Park, NJ); and (10) linoleoyl polyoxyl-6 glycerides NF sold under the brand name Labrafil® M2125CS CC (Gattefosse Corp. Paramus, NJ).

Table S

The prototypes performance was tested by quantitative dispersion study using Fiberoptic dissolution method. The formulations containing were filled into air-filled capsules with a fill weight of about 1g. Pure crystallized ErSO API (30 mg) was also tested as the control. Fasted state small intestinal conditions fluid (“FaSSIF”) (pH = 6.5) and simulated gastric fluid (“SGF”) (pH = 1 .3) were used as the dissolution media. The media volume was 500 ml, stirred at 75 rpm with paddle (USP Apparatus II). The total drug load for each prototype was 30 mg. PDPK analysis indicated that the solubility of ErSO needs to be maintained in both upper and lower gastrointestinal track at different pHs in order to improve bioavailability. The solubility of the ErSO API was monitored continuously over a 12-hour period to ensure that the formulations were able to maintain the API solubility.

From the SGF screen, all the prototypes performed better than the native ErSO API. However, prototypes A6 and A9 did not achieve the maximum solubility enhancement of the ErSO API in acidic condition that mimics the stomach pH. The other prototypes, A5, A7, A8 and A10, were able to reach about 100% solubility and maintain the solubility during the 12- hour period, as shown in Figure 14A. Therefore, A5, A7, A8 and Al 0 were selected to screen in FaSSIF media to ensure the solubility was also enhanced in a higher pH environment. Figure 14B shows the quantitative dispersion of the prototypes in FaSSIF media at pH = 6.5. All the prototypes were dispersed and reached their maximum solubility within an hour compared to the long dissolving time with just the pure ErSO API. After a 12-hour period, pure ErSO API and A5 prototype reached the same level of solubility. The other 3 prototypes were able to reach the 100% level and maintained the solubility throughout the testing period.

EXAMPLE 3

Physical stability was performed on the A7, A8 and A10 formulations from Example 2. The formulations were held at the room temperatures for 7 days with observations made at not less than 3 time points. The observations of the initial assessment of the physical stability of these three formulations are presented in Table 6 below.

Table 6

The three formulations were stable at room temperature physically. No precipitation was observed during the testing period. Small phase separation was observed in the AW formulation.

EXAMPLE 4

Pharmacokinectic profiles of ErSO following capsule dosing of micronized ErSO API and the A7, A8 and A10 formulations from Example 2 in three female beagle dogs were determined as follows. All three dogs received the four prototypes in the following order:

1 . Group 1 received the micronized ErSO API;

2. Group 2 received the A7 formulation; 3. Group 3 received the A8 formulation; and

4. Group 4 received the AI D formulation.

The dose level ErSO in each prototype was 30 mg/animal and the dose formulation concentration was 15 mg/g. Blood samples of approximately 1.0 mL were collected at 0.25 hr, within ± 1 minutes; 0.5 hr, within ± 2 minutes; 1-4 hr, within ± 5 minutes; 8-12 hr, within ± 10 minutes; and 24 hr, within ± 30 minutes. PK plasma samples were analyzed using LC- MS/MS method. One dog was removed from the analysis.

The PK data for each group was used to calculate the effective dose of ErSO obtained from the oral administration of a 30 mg dose of ErSO for each prototype. For micronized ErSO API, the effective dose was determined to be 5 mg, shown in Figure 15A, For formulation A7, the effective dose was determined to be 13 mg, shown in Figure 15B. For formulation A8, the effective dose was determined to be 4 mg, shown in Figure 15C. For formulation A10, the effective dose was determined to be 8 mg, shown in Figure 15D.

The highest concentration of ErSO found in the blood for each prototype (C_max) was 121 ng/ml for the micronized ErSO API; 618 ng/ml for the A7 formulation; 149 ng/ml for the A8 formulation; and 233 ng/ml for the A10 formulation, as shown In Figure 14C. The area under the curve from the time of dosing to the time of last observation at 24 hours (AUC0.24) was 802 ng*h/ml for the micronized ErSO API; 2081 ng*h/ml for the A7 formulation; 661 ng*h/mi for the A8 formulation; and 1376 ng*h/ml for the A10 formulation. These drug exposure results presented in Figure 14C showed good correlation with the FaSSIF solubility results shown in Figure 14B.I

Table 7 below compares the tested formulations for effective dose (Dose best fit), percentage of dose loss (Dose loss), and dose exposure including total amount of drug absorbed by the body (Fo-24 (%), DN Cmax and DN AUC0-24) and total drug exposure across time (Fo-inf (%)). Abbreviations: DN: Dose normalized to 30 mg. Fa: predicted fraction of formulation absorbed. Fo-24 (%): fraction of formulation absorbed over 24 hours. F₀-inf (%): fraction of formulation absorbed from 0 to infinity. Cmax: highest concentration of ErSO found in the blood for each prototype. AUC0.24: Area under the curve from the time of dosing to the time of last observation at 24 hours that is greater than the limit of quantitation. Table 7

Formulation A7 performed the best of the four prototypes. As the fraction absorbed for all formulations and doses tested was predicted to be 100%, the lower dose loss for formulation A7 may have been due to a higher protection against ErSO API degradation in the gastrointestinal tract. * * * * * * *

All publications, patents, and patent documents are incorporated by reference herein, as though individually incorporated by reference. This statement of incorporation by reference is intended by applicants, pursuant to 37 C.F.R. §1 -57(b)(1), to relate to each and every individual publication, patent application, or patent, each of which is clearly identified in compliance with 37 C.F.R. §1.57(b)(2), even if such citation is not immediately adjacent to a dedicated statement of incorporation by reference. The inclusion of dedicated statements of incorporation by reference, if any, within the specification does not in any way weaken this general statement of incorporation by reference. Citation of the references herein is not intended as an admission that the reference is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents. No limitations inconsistent with this disclosure are to be understood therefrom.

The Invention has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many variations and modifications may be made while remaining within the spirit and scope of the invention.

While specific embodiments have been described above with reference to the disclosed embodiments and examples, such embodiments are only illustrative and do not limit the scope of the invention. Changes and modifications can be made in accordance with ordinary skill in the art without departing from the invention in its broader aspects as defined in the following claims. Such equivalents are intended to be encompassed by the following claims. Exemplary methods and uses are set out in the foiiowmg items:

Item 1 A method of ameliorating or treating an ERa positive cancer in a human patient, comprising administering to the human patient a compound having the formula:

once a week in a therapeutically effective amount greater than 1 .5 mg/kg of body weight when administered orally or in a therapeutically effective amount of at least 0.4 mg/kg of body weight when administered intravenously, wherein the weekly dose is repeated until there is complete regression of the ERa positive cancer.

Item 2. The method of item 1 , wherein the compound is administered orally in a therapeutically effective amount of 1 .6 to 4.9 mg/kg of body weight.

Item 3. The method of item 1 , wherein the compound is administered orally in a therapeutically effective amount of 1 .6 to 4.1 mg/kg of body weight.

Item 4. The method of item 1 , wherein the compound is administered orally in a therapeutically effective amount of 1 .6 to 3.3 mg/kg of body weight. Item 5. The method of item 1 , wherein the compound is administered orally in a therapeutically effective amount of 2.0 to 3.3 mg/kg of body weight.

Item 6. The method of item 1 , wherein the compound is administered orally in a therapeutically effective amount of 2.4 to 3.3 mg/kg of body weight. item 7. The method of Item 1 , wherein the compound is administered orally in a therapeutically effective amount of 3.3 mg/kg of body weight.

Item 8. The method of item 1 , wherein the compound is administered intravenously in a therapeutically effective amount of 0.4 to less than 1 .6 mg/kg of body weight.

Item 9. The method of item 1 , wherein the compound is administered intravenously in a therapeutically effective amount of 0.4 to 0.8 mg/kg of body weight. tern 10. The method of item 1 , wherein the weekly dose of the compound is repeated until growth of the ERa positive cancer is inhibited.

Item 1 1 . The method of item 1 , wherein the ERa positive cancer is a breast cancer, an ovarian cancer, a uterine cancer, a cervical carcinoma, or an endometrial cancer. Item 12. A compound having the formula

for use in a method of ameliorating or treating an ERa positive cancer in a human patient, wherein the method comprises administering the compound to the human patient once a week in a therapeutically effective amount greater than 1 .5 mg/kg of body weight when administered orally or a therapeutically effective amount of at least 0.4 mg/kg of body weight when administered intravenously, wherein the weekly dose is repeated until there is complete regression of the ERa positive cancer.

Item 13. The compound of item 12, wherein the therapeutically effective amount of the compound administered orally is 1 .6 to 4.9 mg/kg of body weight. Item 14. The compound of item 12, wherein the therapeutically effective amount of the compound administered orally is 1 .6 to 4.1 mg/kg of body weight. item 15. The compound of item 12, wherein the therapeutically effective amount of the compound administered orally is 1 .6 to 3.3 mg/kg of body weight. item 16. The compound of item 12, wherein the therapeutically effective amount of the compound administered orally is 2.0 to 3.3 mg/kg of body weight. item 17. The compound of item 12, wherein the therapeutically effective amount of the compound administered orally is 2.4 to 3.3 mg/kg of body weight.

Item 18, The compound of item 12, wherein the therapeutically effective amount of the compound administered orally is 3.3 mg/kg of body weight. tern 19. The compound of item 12, wherein the therapeutically effective amount of the compound administered intravenously 0.4 to less than 1 .6 mg/kg of body weight.

Item 20. The compound of item 12, wherein the therapeutically effective amount of the compound administered intravenously 0.4 to 0.8 mg/kg of body weight.

Item 21 . The compound of item 12, wherein the weekly dose is repeated until growth of the ERa positive cancer is inhibited.

Item 22. The compound of tern 12, wherein the ERa positive cancer is a breast cancer, an ovarian cancer, a uterine cancer, a cervical carcinoma, or an endometrial cancer.

Item 23. A method of inhibiting growth of an ERa positive tumor in a mammal, comprising administering to the mammal a compound having the formula

once a week in an amount greater than 20 mg/kg of body weight when administered orally or in an amount of at least 5 mg/kg of body weight when administered intravenously, wherein the weekly dose is repeated until growth of the ERa positive tumor is inhibited.

Item 24. The method of item 23, wherein the compound is administered orally in an amount of 20 to 60 mg/kg of body weight.

Item 25. The method of item 23, wherein the compound is administered orally in an amount of 20 to 50 mg/kg of body weight.

Item 26, The method of item 23, wherein the compound is administered orally in an amount of 20 to 40 mg/kg of body weight.

Item 27. The method of item 23, wherein the compound is administered orally in an amount of 25 to 40 mg/kg of body weight.

Item 28. The method of item 23, wherein the compound is administered orally in an amount of 30 to 40 mg/kg of body weight. tern 29. The method of item 23, wherein the compound is administered orally in an amount of 40 mg/kg of body weight. item 30. The method of item 23, wherein the compound is administered intravenously in an amount of 5 to less than 20 mg/kg of body weight.

Item 31. The method of item 23, wherein the compound is administered intravenously in an amount of 5 to 10 mg/kg of body weight.

Item 32, The method of item 33, wherein the ERa positive tumor is from a breast cancer, ovarian cancer, uterine cancer, cervical carcinoma, or endometrial cancer.

Item 33: A formulation of a compound having the formula

comprising the compound in an amount of 3% w/w, polyethylene glycol 400 in an amount of 40% w/w, vitamin E TPGS in an amount of 20% w/w, and caprylocaproyi polyoxyl-8 glycerides in an amount of 37% w/w.

Item 34: A formulation of a compound having the formula

comprising the compound in an amount of 3% w/w, olive oil in an amount of 47% w/w, propylene glycol monolaurate in an amount of 10% w/w, lecithin in an amount of 10% w/w, and either polysorbate 80 or lauroyl polyoxyl-32 glycerides in an amount of 30% w/w.

Claims

What is claimed:

1. A method of ameliorating or treating an ERa positive cancer in a human patient, comprising administering to the human patient a compound having the formula

2. The method of claim 1 , wherein the compound is administered orally in a therapeutically effective amount of 1 .6 to 4.9 mg/kg of body weight.

3. The method of claim 1 , wherein the compound is administered orally in a therapeutically effective amount of 1 .6 to 4.1 mg/kg of body weight.

4. The method of claim 1 , wherein the compound is administered orally in a therapeutically effective amount of 1 .6 to 3.3 mg/kg of body weight.

5. The method of claim 1 , wherein the compound is administered orally in a therapeutically effective amount of 2.0 to 3.3 mg/kg of body weight.

6. The method of claim 1 , wherein the compound is administered orally in a therapeutically effective amount of 2.4 to 3.3 mg/kg of body weight.

7. The method of claim 1 , wherein the compound is administered orally in a therapeutically effective amount of 3.3 mg/kg of body weight.

8. The method of claim 1 , wherein the compound is administered intravenously in a therapeutically effective amount of 0.4 to less than 1 .6 mg/kg of body weight.

9. The method of claim 1 , wherein the compound is administered intravenously in a therapeutically effective amount of 0.4 to 0.8 mg/kg of body weight.

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10. The method of claim 1 , wherein the weekly dose of the compound is repeated until growth of the ERa positive cancer is inhibited.

11 . The method of claim 1 , wherein the ERa positive cancer is a breast cancer, an ovarian cancer, a uterine cancer, a cervical carcinoma, or an endometrial cancer.

12. A compound having the formula

for use in a method of ameliorating or treating an ERa positive cancer in a human patient, wherein the method comprises administering the compound to the human patient once a week In a therapeutically effective amount greater than 1.5 mg/kg of body weight when administered orally or a therapeutically effective amount of at least 0.4 mg/kg of body weight when administered intravenously, wherein the weekly dose is repeated until there is complete regression of the ERa positive cancer.

13. The compound of claim 12, wherein the therapeutically effective amount of the compound administered orally is 1 .6 to 4.9 mg/kg of body weight.

14. The compound of claim 12, wherein the therapeutically effective amount of the compound administered orally is 1 .6 to 4.1 mg/kg of body weight.

15. The compound of claim 12, wherein the therapeutically effective amount of the compound administered orally is 1 .6 to 3.3 mg/kg of body weight.

16. The compound of claim 12, wherein the therapeutically effective amount of the compound administered orally is 2.0 to 3.3 mg/kg of body weight.

17. The compound of claim 12, wherein the therapeutically effective amount of the compound administered orally is 2.4 to 3.3 mg/kg of body weight.

18. The compound of claim 12, wherein the therapeutically effective amount of the compound administered orally is 3.3 mg/kg of body weight.

19. The compound of claim 12, wherein the therapeutically effective amount of the compound administered intravenously 0.4 to less than 1 .6 mg/kg of body weight,

20. The compound of claim 12, wherein the therapeutically effective amount of the compound administered intravenously 0.4 to 0.8 mg/kg of body weight.

21. The compound of claim 12, wherein the weekly dose is repeated until growth of the ERa positive cancer is inhibited.

22. The compound of claim 12, wherein the ERa positive cancer is a breast cancer, an ovarian cancer, a uterine cancer, a cervical carcinoma, or an endometrial cancer.

23. A method of inhibiting growth of an ERa positive tumor in a mammal, comprising administering to the mammal a compound having the formula

24. The method of claim 23, wherein the compound is administered orally in an amount of 20 to 60 mg/kg of body weight.

25. The method of claim 23, wherein the compound is administered orally in an amount of 20 to 50 mg/kg of body weight.

26. The method of claim 23, wherein the compound is administered orally in an amount of 20 to 40 mg/kg of body weight.

27. The method of claim 23, wherein the compound is administered orally in an amount of 25 to 40 mg/kg of body weight.

28. The method of claim 23, wherein the compound is administered orally in an amount of 30 to 40 mg/kg of body weight.

29. The method of claim 23, wherein the compound is administered orally in an amount of 40 mg/kg of body weight.

30. The method of claim 23, wherein the compound is administered intravenously in an amount of 5 to less than 20 mg/kg of body weight.

31 . The method of claim 23, wherein the compound is administered intravenously in an amount of 5 to 10 mg/kg of body weight.

32. The method of claim 23, wherein the ERa positive tumor is from a breast cancer, ovarian cancer, uterine cancer, cervical carcinoma, or endometrial cancer.

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