Vitamin d analogs and methods of treating cancer using the same

Abstract

The present disclosure provides novel Vitamin D analogs, pharmaceutical compositions containing such compounds, and methods for using the compounds as therapeutic agents. These compounds may be useful in the treatment of cancer, particularly melanoma, ovarian, and breast cancers. Also provided herein are methods of treating cancers by administering at least one compound disclosed herein and at least one additional therapy.

Description

VITAMIN D ANALOGS AND METHODS OF TREATING CANCER USING THESAMETECHNICAL FIELD

[0001] This disclosure relates to novel vitamin D analogs and methods of using and pharmaceutical compositions comprising a compound having the chemical structure:BACKGROUND

[0002] Breast and ovarian cancers are two of the most common types of cancer that affect women. According to the CDC, ovarian cancer is the second most common gynecologic cancer in the United States, causing more deaths than any other cancer of the female reproductive system. The National Cancer Institute reports that about 13% of women in the U.S. population will develop breast cancer during their lives.

[0003] The incidence rate of ovarian cancer in the U.S. has been slowly falling over the past 20 years, declining by 1-2% per year from 1990 to the mid-2010s and by almost 3% per year from 2015 to 2019. This is likely due to more use of oral contraceptives and less use of hormonal treatment for the symptoms of menopause.

[0004] The National Cancer Institute reports that a woman’s lifetime risk of developing breast and / or ovarian cancer is markedly increased if she inherits a harmful variant in BRCA1 or BRCA2. Women with BRCA1 mutations have a 40 percent to 60 percent chance of being diagnosed with ovarian cancer, while those with BRCA2 mutations have a 20 percent to 35 percent lifetime risk. BRCA1 mutations lead to a 55 percent to 72 percent risk of developing breast cancer.

[0005] In studies of cancer cells and of tumors in mice, vitamin D has been found to have several activities that might slow or prevent the development of cancer, including promoting cellular differentiation, decreasing cancer cell growth, stimulating apoptosis, and reducing angiogenesis. During the study of vitamin D intake on the effects of bone health or other non-cancer outcomes it was discovered that incidence and mortality rate ofcolorectal cancer, breast cancer, prostate cancer, and pancreatic cancer decreased in subjects with higher intake, or blood levels, of vitamin D. Previously reported vitamin D analogs have exhibited efficacy in animal trials for breast, ovarian, prostate, and skin cancer, albeit with evidence of dose-limiting adverse effects, including renal toxicities such as nephrocalcinosis. There remains a need for effective anticancer therapies for the treatment of breast, ovarian and skin cancers. New vitamin D analogs that reduce or eliminate the toxicities of the compounds previously shown to be effective in the treatment of these cancers, could partially fulfill this need.SUMMARY

[0006] Disclosed herein are novel organic compounds and pharmaceutical compositions containing such compounds. These compounds may be useful in the treatment of cancer.

[0007] This disclosure also provides pharmaceutical compositions comprising these analogs of Vitamin D and a pharmaceutically acceptable carrier.

[0008] This disclosure also provides methods of treating diseases and disorders utilizing these novel analogs of Vitamin D and pharmaceutically acceptable salts, hydrates, solvates, clathrates, and polymorphs thereof.

[0009] In these compositions and methods of use, the Vitamin D analog may be (lS,5R,E)-5-hydroxy-3-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a- dimethyldihydro-lH-inden-4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-2- methylenecyclohexyl 2-bromoacetate (hereinafter “Compound A”), which is a compound of Formula I, and which has the chemical structure:

[0010] This compound is a geometric isomer (i.e., stereoisomer or geometric “E” isomer) of (lR,3S,Z)-3-hydroxy-5-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)- 3 a, 7 a-dimethyldihy dro- 1 H-inden-4(2H, 5H, 6H,7H,7aH)-ylidene)ethylidene)-4-methylenecyclohexyl 2-bromoacetate (FIG. 1 A), disclosed in the inventors’ copending application No. 18 / 012,975 (U.S. Patent Pub. No. 2023 / 0241081), which is incorporated herein in its entirety. The inventors have made the surprising and unexpected discovery that this “E” isomer (Compound A; FIG. IB) advantageously maintains nearly the same or greater anti-cancer activity while displaying substantially reduced systemic toxicities or “off-target” adverse effects compared to the previously disclosed “Z” isomer.

[0011] In the compositions and methods of use provided in this disclosure, the novel Vitamin D analogs are substantially or completely free of the previously disclosed Z stereomeric forms.

[0012] This invention particularly relates to the E stereoisomer, Compound A (FIG. IB), which is (1 S,5R,E)-5-hydroxy-3-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2- yl)-3a,7a-dimethyldihydro-lH-inden-4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-2- methylenecyclohexyl 2-bromoacetate, which is believed to have substantially reduced toxicity and other benefits as compared to its Z stereoisomer: (lR,3S,Z)-3-hydroxy-5-((E)- 2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a-dimethyldihydro-lH-inden- 4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-4-methylenecyclohexyl 2-bromoacetate.

[0013] This disclosure encompasses the use of the novel Vitamin D analogs of formula I for treating or preventing cancers, including, but not limited to cancer of the head, thyroid, neck, eye, skin, mouth, throat, esophagus, chest, bone, blood, bone marrow, lung, colon, sigmoid, rectum, stomach, prostate, breast, ovaries, kidney, liver, pancreas, brain, intestine, heart, adrenal, subcutaneous tissue, lymph nodes, heart, and combinations thereof. Specific cancers that can be treated by this method are ovarian cancer, malignant melanoma, and triple-negative breast cancer.

[0014] This disclosure further encompasses pharmaceutical compositions and single unit dosage forms comprising the novel Vitamin D analogs of formula I and pharmaceutically acceptable polymorphs, salts, hydrates, clathrates, and solvates thereof. These pharmaceutical compositions may be substantially or completely free of the previously disclosed Z stereomeric forms.BRIEF DESCRIPTION OF THE FIGURES

[0015] FIG. 1 A shows the chemical structure of the previously disclosed Z isomer: (lR,3S,Z)-3-hydroxy-5-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a- dimethyldihydro- lH-inden-4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-4- methylenecyclohexyl 2-bromoacetate.

[0016] FIG. IB illustrates the chemical structure of (lS,5R,E)-5-hydroxy-3-((E)-2- ((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a-dimethyldihydro-lH-inden- 4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-2-methylenecyclohexyl 2-bromoacetate (Compound A of this disclosure).

[0017] FIG. 2 depicts a chemical synthesis scheme for the synthesis of Compound A [(lS,5R,E)-5-hydroxy-3-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a- dimethyldihydro- lH-inden-4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-2- methylenecyclohexyl 2-bromoacetate] .

[0018] FIGS. 3 A, 3B, and 3C show the growth in tumor volume in a mouse xenograft model in mice treated with Compound A or control (vehicle). FIG. 3 A shows the inhibition of tumor growth in a xenograft model of UCD46 triple negative breast cancer patient- derived xenograft (PDX) cells, and the percent change in tumor volume at day 45. FIG. 3B shows the inhibition of tumor growth in a xenograft model of UCD49 TNBC cells derived from a PDX, and the percent change in tumor volume at day 28. FIG. 3C shows the inhibition of tumor growth in a xenograft model of MDA-MB-231 TNBC cells and the percent change in tumor volume at day 19.

[0019] FIG. 4 is a graph of the weight of mice treated with escalating doses of a compound of this disclosure or escalating doses of a previously disclosed isomer of a compound of this disclosure.

[0020] FIG. 5 is a bar graph of the serum calcium concentrations of mice treated with two different doses of a compound of this disclosure or escalating doses of a previously disclosed isomer of a compound of this disclosure.

[0021] FIGS. 6A and 6B are bar graphs of the gene expression of CYP24 relative to GAPDH gene expression in melanoma cells (FIG. 6A) or ovarian cancer cells (FIG. 6B) treated with vitamin D3, or a compound of this disclosure, or a previously disclosed isomer of a compound of this disclosure.

[0022] FIG. 7 and FIG. 8 are photomicrographs of renal tissue samples taken from mice treated with a compound of this disclosure or a previously disclosed isomer of a compound of this disclosure, stained with H&E stain (FIG. 7) or Von Kosa stain (FIG. 8).

[0023] FIG. 9 shows the growth inhibition in human ovarian carcinoma cells (OVCAR3 cell line) in an in vitro model treated with Compound A or control (vehicle) and the final tumor cell confluence following treatment in the two cell groups.

[0024] FIG. 10 shows the growth of LOX melanoma tumor volume in a mouse xenograft model in mice treated with Compound A or control (vehicle) and the final tumor volumes in the two treatment groups.

[0025] FIG. 11 shows the growth of MB4667 melanoma cell tumor volume in a mouse xenograft model in mice treated with Compound A or control (vehicle), and the final tumor volumes in the two treatment groups.DEFINITIONS

[0026] As used herein, term “Formula I” refers to the stereomerically pure “E” isomeric form of compounds having the chemical structure:wherein: X1is H, OH, or -OC(=O)CH2X3;X2is H or OH; and,X3is F, Cl, Br, or I.

[0027] As used herein, the term “Compound A” refers to stereomerically pure (1S,5R,E)-5-hydroxy-3-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a- dimethyldihydro-lH-inden-4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-2- methylenecyclohexyl 2-bromoacetate (FIG. IB).

[0028] Compound A has the following chemical structure:

[0029] As used herein, the term “patient” refers to a mammal, particularly a human, in need of cancer treatment.

[0030] As used herein, the term “pharmaceutically acceptable salts” refers to non-toxic salts of the compounds of this disclosure (see, J. Pharm. Set., 1997, 66(1): 1-19). Other salts may, however, be useful in the preparation of compounds of this disclosure or of their pharmaceutically acceptable salts.

[0031] As used herein and unless otherwise indicated, the term “stereomerically pure” means a composition that comprises one stereoisomer of a compound and is substantially free of other stereoisomers of that compound. For example, a stereomerically pure composition of a compound having one chiral center will be substantially free of the opposite enantiomer of the compound. A stereomerically pure composition of a compound having two chiral centers will be substantially free of other diastereomers of the compound. Atypical stereomerically pure compound comprises greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomers of the compound, even more preferably, greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomers of the compound, and most preferably greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomers of the compound.

[0032] As used herein, the terms “adverse effects” or “toxicities” includes, but is not limited to, lethargy, weight loss, elevated serum calcium (hypercalcemia), gastrointestinal and renal toxicities, including renal calcium deposits, nephrocalcinosis. Vitamin D toxicity similarly manifests from hypercalcemia's effects. Often, symptoms can be nonspecific and subtle, such as weakness, fatigue, anorexia, and bone pains. More severe symptoms include neurological symptoms including confusion, apathy, agitation, irritability, andsometimes, ataxia, stupor, and coma. Gastrointestinal symptoms include abdominal pain, nausea, vomiting, constipation, peptic ulcers, and pancreatitis (from malignant calcifications). Renal symptoms manifest as polyuria, polydipsia, and nephrolithiasis. Severe hypercalcemia can also lead to cardiac arrhythmias. Clinical signs of toxicity in patients may include loss of skin turgor and dry mucous membranes (due to dehydration); changes in mental status; and abdominal tenderness without rebound, rigidity, or guarding. All of these adverse effects and signs of toxicity may be seen in mammals following administration of the previously disclosed compound [(lR,3S,Z)-3-hydroxy-5-((E)-2- ((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a-dimethyldihydro-lH-inden- 4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-4-methylenecyclohexyl 2-bromoacetate], and are significantly reduced or eliminated following the administration of a compound of this disclosure, such as Compound A.

[0033] As used herein and unless otherwise indicated, the phrases “reduce or avoid adverse effects” and “reducing or avoiding adverse effects” “reducing or avoiding toxicities” mean the reduction of the severity of one or more adverse effects or toxicities as defined herein.

[0034] It should be noted that if there is a discrepancy between a depicted chemical structure and a name given that structure, the depicted structure is to be accorded more weight. In addition, if the stereochemistry of a structure or a portion of a structure is not indicated, the structure or portion of the structure is to be interpreted as encompassing all stereoisomers at that position or within the molecule.DETAILED DESCRIPTION

[0035] As conventional vitamin D analogs exert their activity through modulation of vitamin D receptor activity, it was highly unexpected and extremely surprising to discover that compounds of Formula I exerted an anti-cancer efficacy independent of the vitamin D receptor and that the E stereoisomer displayed the same or greater anti-cancer activity while causing substantially lower adverse effects.

[0036] This disclosure provides these stereomerically pure forms of the compounds of Formula I, specifically including stereomerically pure Compound A, substantially free of its other, “Z” stereoisomer, as well as novel methods using these compounds, and pharmaceutical compositions comprising the stereomerically pure forms of these compounds.

[0037] For example, this disclosure provides the in vitro and in vivo use of the compounds of Formula I, and the incorporation of one or more of these compounds into pharmaceutical compositions and single unit dosage forms useful in the treatment and prevention of a variety of cancers. Specific methods of this disclosure reduce or avoid the adverse effects associated with Vitamin D or derivatives of Vitamin D and related anticancer analogs. Other specific methods of this disclosure reduce or avoid the adverse effects associated with the use of the previously disclosed (lR,3S,Z)-3-hydroxy-5-((E)-2- ((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a-dimethyldihydro-lH-inden- 4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-4-methylenecyclohexyl 2-bromoacetate.

[0038] Specific methods of this disclosure include methods of treating or preventing diseases and disorders including, but not limited to, solid tumor cancers and blood-born cancers, including specifically, ovarian cancer, malignant melanoma, and triple-negative breast cancer.

[0039] Pharmaceutical compositions and dosage forms of this disclosure, which may comprise compounds of Formula I, such as Compound A, or a pharmaceutically acceptable polymorph, salt, clathrate, solvate, or hydrate thereof, can be used in the methods of this disclosure.

[0040] This disclosure also provides methods of treating or preventing cancer, including but not limited to, solid tumors, blood-born tumors, and in particular, ovarian cancer, malignant melanoma, and triple-negative breast cancer in a patient, which comprises administering to a patient in need of such treatment or prevention a therapeutically effective amount of a stereomerically pure compound of Formula I, or a pharmaceutically acceptable metabolite, polymorph, salt, solvate, hydrate, or clathrate thereof; in particular wherein the patient is a mammal, in particular wherein the patient is a human.

[0041] Stereomerically pure compounds of Formula I, or a pharmaceutically acceptable polymorph, salt, solvate, hydrate, or clathrate thereof, may be adjunctively administered with at least one additional therapeutic agent. Examples of additional therapeutic agents include, but are not limited to, anti-cancer drugs, anti-inflammatories, antihistamines, and decongestants. Specific additional therapeutic agents include but are not limited to aldesleukin, binimetinib, encorafenib, cobimetinib, dabrafenib mesylate, dacarbazine, doxorubicin, encorafenib, aldesleukin (IL-2), paclitaxel, talimogene laherparepvec, recombinant interferon a-2b, peginterferon a-2b, ipilimumab, pembrolizumab, trametinib, binimetinib, nivolumab, vemurafenib, tebentafusp, or combinations thereof.Administration of one or more of these additional compounds may be carried out simultaneously, in tandem, or at different treatment regimens.

[0042] Methods of Treatment

[0043] This disclosure encompasses methods of treating and preventing cancer in a patient which comprise administering to a patient in need of such treatment or prevention a therapeutically effective amount of a stereomerically pure compound of Formula I, or a pharmaceutically acceptable metabolite, polymorph, salt, solvate, hydrate, or clathrate thereof. In particular, these methods include treating and / or preventing ovarian cancer, malignant melanoma, or triple-negative breast cancer in a patient.

[0044] Specific methods of this disclosure avoid or reduce toxicities or adverse effects associated with agents used in the treatment of such cancers, including toxicities or adverse effects caused by the administration of (lR,3S,Z)-3-hydroxy-5-((E)-2-((3aS,7aR)- l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a-dimethyldihydro-lH-inden- 4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-4-methylenecyclohexyl 2-bromoacetate.

[0045] As stated above, the active compounds of this disclosure (i.e., compounds of Formula I) may be used in the treatment or prevention of a wide range of diseases and conditions. The magnitude of a prophylactic or therapeutic dose of a particular active ingredient of this disclosure in the acute or chronic management of a disease or condition will vary, however, with the nature and severity of the disease or condition, and the route by which the active ingredient is administered. The dose, and perhaps the dose frequency, will also vary according to the age, body weight, and response of the individual patient. Suitable dosing regimens can be readily selected by those skilled in the art with due consideration of such factors. In general, the recommended daily dose range for the conditions described herein lie within the range of from about 1 mg to about 1000 mg per day.

[0046] It may be necessary to use dosages of the active ingredient outside the ranges disclosed herein in some cases, as will be apparent to those of ordinary skill in the art. Furthermore, it is noted that the clinician or treating physician will know how and when to interrupt, adjust, or terminate therapy in conjunction with individual patient response.

[0047] The phrases “therapeutically effective amount”, “ prophy lactically effective amount” and “therapeutically or prophylactically effective amount,” as used herein encompasses the above-described dosage amounts and dose frequency schedules. Different therapeutically effective amounts may be applicable for different diseases andconditions, as will be readily known by those of ordinary skill in the art. Similarly, amounts sufficient to treat or prevent such disorders, but insufficient to cause, or sufficient to reduce, adverse effects associated with (lR,3S,Z)-3-hydroxy-5-((E)-2-((3aS,7aR)-l- ((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a-dimethyldihydro-lH-inden- 4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-4-methylenecyclohexyl 2-bromoacetate are also encompassed by the above described dosage amounts and dose frequency schedules.

[0048] This disclosure also provides methods of treating cancers, such as breast, ovarian, or skin cancers, using combination therapy. The co-administration of the other oncolytic therapies can be carried out simultaneously, in tandem, or at different treatment regimens.

[0049] In these methods, the combination therapy may include one or more immunotherapies. Such combinations can be carried out simultaneously, in tandem, or at different treatment regimens. These immunotherapies may comprise a PD-1 inhibitor or a PD-L1 inhibitor.

[0050] Pharmaceutical Compositions

[0051] Pharmaceutical compositions and single unit dosage forms comprising one or more compounds of Formula I, such as Compound A, or a pharmaceutically acceptable polymorph, salt, solvate, hydrate, or clathrate thereof, are encompassed by this disclosure. Individual dosage forms of this disclosure may be suitable for oral, mucosal (including rectal, nasal, or vaginal), parenteral (including subcutaneous, intramuscular, bolus injection, intraarterial, or intravenous), sublingual, transdermal, buccal, or topical administration. Pharmaceutical compositions and dosage forms of this disclosure typically also comprise one or more pharmaceutically acceptable excipients.

[0052] The pharmaceutical compositions of this disclosure may also encompass at least one additional therapeutic agent. Exemplary additional therapeutic agents include, but are not limited to: aldesleukin, binimetinib, encorafenib, cobimetinib, dabrafenib mesylate, dacarbazine, doxorubicin, encorafenib, aldesleukin (IL-2), paclitaxel, talimogene laherparepvec, recombinant interferon a-2b, peginterferon a-2b, ipilimumab, pembrolizumab, trametinib, binimetinib, nivolumab, vemurafenib, tebentafusp, or combinations thereof.

[0053] Single unit dosage forms of this disclosure are suitable for oral, mucosal (e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g., subcutaneous, intravenous, bolus injection, intramuscular, or intraarterial), or transdermal administration to a patient. Examples of dosage forms include, but are not limited to: tablets; caplets; capsules, suchas soft elastic gelatin capsules; cachets; troches; lozenges; dispersions; suppositories; ointments; cataplasms (poultices); pastes; powders; dressings; creams; plasters; solutions; patches; aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage forms suitable for oral or mucosal administration to a patient, including suspensions (e.g. aqueous or nonaqueous liquid suspensions, oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions, and elixirs; liquid dosage forms suitable for parenteral administration to a patient; and sterile solids (e.g., crystalline or amorphous solids) that can be reconstituted to provide liquid dosage forms suitable for parenteral administration to a patient.

[0054] The composition, shape, and type of dosage forms of this disclosure will typically vary depending on their use. For example, a dosage form used in the acute treatment of melanoma or a related cancer may contain larger amounts of one or more of the active ingredients it comprises than a dosage form used in the chronic treatment of the same disease. Similarly, a parenteral dosage form may contain smaller amounts of one or more of the active ingredients it comprises than an oral dosage form used to treat the same disease or disorder. These and other ways in which specific dosage forms encompassed by this invention will vary from one another will be readily apparent to those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton Pa. (1990).

[0055] Typical pharmaceutical compositions and dosage forms comprise one or more excipients. Suitable excipients are well known to those skilled in the art of pharmacy, and non-limiting examples of suitable excipients are provided herein. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on a variety of factors well known in the art including, but not limited to, the way in which the dosage form will be administered to a patient. For example, oral dosage forms such as tablets may contain excipients not suited for use in parenteral dosage forms. The suitability of a particular excipient may also depend on the specific active ingredients in the dosage form.

[0056] Lactose-free compositions of this disclosure can comprise excipients that are well known in the art and are listed, for example, in the U.S. Pharmacopeia (USP) SP (XXI) / NF (XVI). In general, lactose-free compositions comprise an active ingredient, a binder / filler, and a lubricant in pharmaceutically compatible and pharmaceutically acceptable amounts. Preferred lactose-free dosage forms comprise an active ingredient, microcrystalline cellulose, pre-gelatinized starch, and magnesium stearate.

[0057] This invention further encompasses anhydrous pharmaceutical compositions and dosage forms comprising active ingredients, since water can facilitate the degradation of some compounds. For example, the addition of water (e.g., 5%) is widely accepted in the pharmaceutical arts as a means of simulating long-term storage to determine characteristics such as shelf-life or the stability of formulations over time. See, e.g., Jens T. Carstensen, Drug Stability: Principles & Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 1995, pp. 379-80. In effect, water and heat accelerate the decomposition of some compounds. Thus, the effect of water on a formulation can be of great significance since moisture and / or humidity are commonly encountered during manufacture, handling, packaging, storage, shipment, and use of formulations.

[0058] Anhydrous pharmaceutical compositions and dosage forms of this disclosure can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions.

[0059] An anhydrous pharmaceutical composition should be prepared and stored such that its anhydrous nature is maintained. Accordingly, anhydrous compositions are preferably packaged using materials known to prevent exposure to water such that they can be included in suitable formulary kits. Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastics, unit dose containers (e.g., vials), blister packs, and strip packs.

[0060] This disclosure further encompasses pharmaceutical compositions and dosage forms that comprise one or more compounds that reduce the rate at which an active ingredient will decompose. Such compounds, which are referred to herein as “stabilizers,” include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers.

[0061] Like the amounts and types of excipients, the amounts and specific types of active ingredients in a dosage form may differ depending on factors such as, but not limited to, the route by which it is to be administered to patients. However, typical dosage forms of this disclosure comprise compounds of Formula I, such as Compound A, or a pharmaceutically acceptable salt, solvate, clathrate, hydrate, or polymoprh thereof lie within the range of from about 1 mg to about 1000 mg per day, given as a single once-a- day dose in the morning but preferably as divided doses throughout the day taken with food. More specifically, the daily dose is administered twice daily in equally divided doses. Specifically, a daily dose range should be from about 5 mg to about 500 mg perday, more specifically, between about 10 mg and about 200 mg per day. In managing the patient, the therapy should be initiated at a lower dose, perhaps about 1 mg to about 25 mg, and increased if necessary up to about 200 mg to about 1000 mg per day as either a single dose or divided doses, depending on the patient's global response.

[0062] Oral Dosage Forms

[0063] Pharmaceutical compositions of this disclosure that are suitable for oral administration can be presented as discrete dosage forms, such as, but are not limited to, tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g., flavored syrups). Such dosage forms contain predetermined amounts of active ingredients, and may be prepared by methods of pharmacy well known to those skilled in the art. See generally, Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton Pa. (1990).

[0064] Typical oral dosage forms of this disclosure are prepared by combining the active ingredient(s) in an intimate admixture with at least one excipient according to conventional pharmaceutical compounding techniques. Excipients can take a wide variety of forms depending on the form of preparation desired for administration. For example, excipients suitable for use in oral liquid or aerosol dosage forms include, but are not limited to, water, glycols, oils, alcohols, flavoring agents, preservatives and coloring agents. Examples of excipients suitable for use in solid oral dosage forms (e.g., powders, tablets, capsules, and caplets) include, but are not limited to, starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrating agents.

[0065] Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit forms, in which case solid excipients are employed. If desired, tablets can be coated by standard aqueous or nonaqueous techniques. Such dosage forms can be prepared by any of the methods of pharmacy. In general, pharmaceutical compositions and dosage forms are prepared by uniformly and intimately admixing the active ingredients with liquid carriers, finely divided solid carriers, or both, and then shaping the product into the desired presentation if necessary.

[0066] For example, a tablet can be prepared by compression or molding. Compressed tablets can be prepared by compressing in a suitable machine the active ingredients in a free-flowing form such as powder or granules, optionally mixed with an excipient. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0067] Examples of excipients that can be used in oral dosage forms of this disclosure include, but are not limited to, binders, fillers, disintegrants, and lubricants. Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

[0068] Examples of fillers suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof. The binder or filler in pharmaceutical compositions of this disclosure is typically present in from about 50 to about 99 weight percent of the pharmaceutical composition or dosage form.

[0069] Suitable forms of microcrystalline cellulose include, but are not limited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICEL RC-581, AVICEL-PH-105 (available from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. A specific binder is a mixture of microcrystalline cellulose and sodium carboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or low moisture excipients or additives include AVICEL-PH-103™ and Starch 1500 LM.

[0070] Disintegrants are used in the compositions of this disclosure to provide tablets that disintegrate when exposed to an aqueous environment. Tablets that contain too much disintegrant may disintegrate in storage, while those that contain too little may not disintegrate at a desired rate or under the desired conditions. Thus, a sufficient amount of disintegrant that is neither too much nor too little to detrimentally alter the release of the active ingredients should be used to form solid oral dosage forms of this disclosure. The amount of disintegrant used varies based upon the type of formulation and is readily discernible to those of ordinary skill in the art. Typical pharmaceutical compositions comprise from about 0.5 to about 15 weight percent of disintegrant, specifically from about 1 to about 5 weight percent of disintegrant.

[0071] Disintegrants that can be used in pharmaceutical compositions and dosage forms of this disclosure include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.

[0072] Lubricants that can be used in pharmaceutical compositions and dosage forms of this disclosure include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g. peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, and mixtures thereof. Additional lubricants include, for example, a syloid silica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore, Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co. of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold by Cabot Co. of Boston, Mass.), and mixtures thereof. If used at all, lubricants are typically used in an amount of less than about 1 weight percent of the pharmaceutical compositions or dosage forms into which they are incorporated.

[0073] Delayed Release Dosage Forms

[0074] Active ingredients of this disclosure can be administered by controlled release means or by delivery devices that are well known to those of ordinary skill in the art. Examples include, but are not limited to, those described in U.S. Pat. Nos.: 3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548, 5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which is incorporated herein by reference. Such dosage forms can be used to provide slow or controlled-release of one or more active ingredients using, for example, hydropropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or a combination thereof to provide the desired release profile in varying proportions. Suitable controlled-release formulations known to those of ordinary skill in the art, including those described herein, can be readily selected for use with the active ingredients of this disclosure. This disclosure thus encompasses single unit dosage forms suitable for oral administration such as, but not limited to, tablets, capsules, gelcaps, and caplets that are adapted for controlled-release.

[0075] All controlled-release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled counterparts. Ideally, the use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time. Advantages of controlled-release formulations include extended activity of the drug, reduced dosage frequency, and increased patient compliance. In addition, controlled-release formulations can be used to affect the time of onset of action or other characteristics, such as blood levels of the drug, and can thus affect the occurrence of side (e.g., adverse) effects.

[0076] Most controlled-release formulations are designed to initially release an amount of drug (active ingredient) that promptly produces the desired therapeutic effect, and gradually and continually release of other amounts of drug to maintain this level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body. Controlled-release of an active ingredient can be stimulated by various conditions including, but not limited to, pH, temperature, enzymes, water, or other physiological conditions or compounds.

[0077] Parenteral Dosage Forms

[0078] Parenteral dosage forms can be administered to patients by various routes including, but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular, and intraarterial. Because their administration typically bypasses patients’ natural defenses against contaminants, parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions.

[0079] Suitable vehicles that can be used to provide parenteral dosage forms of this disclosure are well known to those skilled in the art. Examples include, but are not limited to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as,but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0080] Compounds that increase the solubility of one or more of the active ingredients disclosed herein can also be incorporated into the parenteral dosage forms of this disclosure.

[0081] Transdermal, Topical, and Mucosal Dosage Forms

[0082] Transdermal, topical, and mucosal dosage forms of this disclosure include, but are not limited to, ophthalmic solutions, sprays, aerosols, creams, lotions, ointments, gels, solutions, emulsions, suspensions, or other forms known to one of skill in the art. See, e.g., Remington's Pharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton Pa. (1980 & 1990); and Introduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger, Philadelphia (1985). Dosage forms suitable for treating mucosal tissues within the oral cavity can be formulated as mouthwashes or as oral gels. Further, transdermal dosage forms include “reservoir type” or “matrix type” patches, which can be applied to the skin and worn for a specific period of time to permit the penetration of a desired amount of active ingredients.

[0083] Suitable excipients (e.g., carriers and diluents) and other materials that can be used to provide transdermal, topical, and mucosal dosage forms encompassed by this invention are well known to those skilled in the pharmaceutical arts, and depend on the particular tissue to which a given pharmaceutical composition or dosage form will be applied. With that fact in mind, typical excipients include, but are not limited to, water, acetone, ethanol, ethylene glycol, propylene glycol, butane-l,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil, and mixtures thereof to form lotions, tinctures, creams, emulsions, gels or ointments, which are non-toxic and pharmaceutically acceptable. Moisturizers or humectants can also be added to pharmaceutical compositions and dosage forms if desired. Examples of such additional ingredients are well known in the art. See, e.g., Remington's Pharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton Pa. (1980 & 1990).

[0084] Depending on the specific tissue to be treated, additional components may be used prior to, in conjunction with, or subsequent to treatment with active ingredients of this disclosure. For example, penetration enhancers can be used to assist in delivering the active ingredients to the tissue. Suitable penetration enhancers include, but are not limited to: acetone; various alcohols such as ethanol, oleyl, and tetrahydrofuryl; alkyl sulfoxidessuch as dimethyl sulfoxide; dimethyl acetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such as polyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; and various water-soluble or insoluble sugar esters such as Tween 80 (polysorbate 80) and Span 60 (sorbitan monostearate).

[0085] The pH of a pharmaceutical composition or dosage form, or of the tissue to which the pharmaceutical composition or dosage form is applied, may also be adjusted to improve delivery of one or more active ingredients. Similarly, the polarity of a solvent carrier, its ionic strength, or tonicity can be adjusted to improve delivery. Compounds such as stearates can also be added to pharmaceutical compositions or dosage forms to advantageously alter the hydrophilicity or lipophilicity of one or more active ingredients so as to improve delivery. In this regard, stearates can serve as a lipid vehicle for the formulation, as an emulsifying agent or surfactant, and as a delivery-enhancing or penetration-enhancing agent. Different salts, hydrates or solvates of the active ingredients can be used to further adjust the properties of the resulting composition.

[0086] Kits

[0087] Active ingredients of this disclosure may not be administered to a patient at the same time or by the same route of administration. This invention therefore encompasses kits which, when used by the medical practitioner, can simplify the administration of appropriate amounts of active ingredients to a patient.

[0088] Atypical kit of this disclosure comprises a unit dosage form of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, clathrate, polymorph or prodrug thereof, and a unit dosage form of a second active ingredient. Examples of additional active ingredients include, but are not limited to, aldesleukin, binimetinib, encorafenib, cobimetinib, dabrafenib mesylate, dacarbazine, doxorubicin, encorafenib, aldesleukin (IL-2), paclitaxel, talimogene laherparepvec, recombinant interferon a-2b, peginterferon a-2b, ipilimumab, pembrolizumab, trametinib, binimetinib, nivolumab, vemurafenib, tebentafusp, or combinations thereof.

[0089] Kits of this disclosure can further comprise devices that are used to administer the active ingredient(s). Examples of such devices include, but are not limited to, syringes, drip bags, patches, and inhalers.

[0090] Kits of this disclosure can further comprise pharmaceutically acceptable vehicles that can be used to administer one or more active ingredients. For example, if an active ingredient is provided in a solid form that must be reconstituted for parenteraladministration, the kit can comprise a sealed container of a suitable vehicle in which the active ingredient can be dissolved to form a particulate-free sterile solution that is suitable for parenteral administration. Examples of pharmaceutically acceptable vehicles include, but are not limited to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.EXAMPLESExample 1: Synthesis of Compound A

[0091] Synthesis of (lS,5R,E)-5-hydroxy-3-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6- methylheptan-2-yl)-3a,7a-dimethyldihydro-lH-inden-4(2H,5H,6H,7H,7aH)- ylidene)ethylidene)-2-methylenecyclohexyl 2-bromoacetate.

[0092] Referring to FIG. 2, a thiophene ring is formed in compound 1 (C27H44O2; molecular weight 400.64) with sulfur dioxide in chloroform to form compound 2 (C27H44O4S; molecular weight 464.70). The exposure of compound 2 to sodium bicarbonate in ethanol forms compound 3 as a mix of the two stereoisomers (C27H44O2; molecular weight 400.64). The desired isomer of compound 3, shown in FIG. 2, is purified from its congener. The silane compound 4 (CssHssChSi; molecular weight 514.90) is formed by contacting compound 3 with triethylsilyl chloride and imidazole. A hydroxyl is formed on the cyclohexane of compound 4 by selenium dioxide-mediated oxidation in N- methylmorpholine-N-oxide to form compound 5 (CssHssChSi; molecular weight 530.90). 2-bromo-2-methylpropanoic acid in N,N’ -di cyclohexylcarbodiimide is used to form a bromo-acetyl ester at the hydroxyl in compound 5 to form compound 6 (CssEfeBrChSi; molecular weight 651.83). Compound A of this disclosure (C29H4sBrO4; molecular weight 537.57) is formed when compound 6 is acidified in hydrofluoric acid.Example 2: Compound A inhibits in vivo growth of triple negative breast cancer tumors

[0093] FIGS. 3 A-3C show the growth inhibition in vivo of triple negative breast cancer by Compound A. Briefly, UCD46 triple negative breast cancer patient-derived xenograft (TNBC PDX) cells (FIG. 3 A), UCD49 TNBC cells derived from a PDX (FIG. 3B), and MDA-MB-231 TNBC cells (FIG. 3C) were used to establish tumors in c57 / B16 mice.Tumor size was measured by caliper and once the tumors reached approximately 100mm3in size, treatments were initiated with intraperitoneal injection of Compound A (0.75 pg / kg) three times weekly. As shown in FIGS. 3A-3C, all reductions in tumor growth in response to Compound A treatments reached significance with p values < 0.05 as determined by one-way ANOVA with Fisher’s LSD post test.Example 3: Substantially lower toxicity of Compound A

[0094] The toxicity of Compound A was investigated in comparison to the adverse effects known to occur following treatment with the previously disclosed Z isomeric form ((lR,3S,Z)-3-hydroxy-5-((E)-2-((3aS,7aR)-l-((R)-6-hydroxy-6-methylheptan-2-yl)-3a,7a- dimethyldihydro-lH-inden-4(2H,5H,6H,7H,7aH)-ylidene)ethylidene)-4- methylenecyclohexyl 2-bromoacetate).

[0095] CD-I mice (Charles River) were administered increasing doses of Compound A or the previously disclosed Z isomer or the vehicle (control). All treatments were done by intraperitoneal injection. Mice weights were monitored daily, and the mice treated with the previously disclosed Z isomer were sacrificed after three days of treatment due to loss of weight and decreased clinical scores (FIG. 4). The mice treated with Compound A maintained weight during the time course and did not display decreases in clinical scores (FIG. 4).

[0096] Blood samples collected from the treated mice and serum calcium levels were measured. Mice treated with the previously disclosed Z isomer showed significantly increased serum calcium levels, whereas the mice treated with Compound A showed no elevation in serum calcium above vehicle-treated control mice (FIG. 5).

[0097] Melanoma cells (FIG. 6A) and ovarian cancer cells (FIG. 6B) were treated with 1,25-dihydroxy vitamin D3 (“1,25D”), Compound A (CpdA), the previously disclosed Z isomer (Z isomer), or vehicle (control) and the gene expression of cytochrome P45024A1 (CYP24A1) was measured relative to GAPDH expression in these cells. As shown in FIGS. 6A and 6B, treatment of these melanoma and ovarian cancer cells with 1,25D and the previously disclosed Z isomer induced significantly increased CYP24A1 gene expression over control-treated cells. In contrast, treatment of these melanoma and ovarian cancer cells with Compound A showed no increase in CYP24A1 gene expression over control -treated cells.

[0098] Kidneys from mice treated with Compound A, or the previously disclosed Z isomer, or vehicle (control) were recovered from the sacrificed animals and analyzed forcalcium deposits, indicative of nephrocalcinosis. As shown in FIG. 7 (hematoxylin and eosin, “H&E” stained; arrows indicate areas of nephrocalcinosis) and FIG8. (Von Kossa stained), renal tissue from the Z isomer treated mice showed numerous calcium deposits, while renal tissue from the Compound A and vehicle-treated mice showed no calcium deposits.

[0099] These data demonstrate that Compound A is considerably less toxic than the previously disclosed Z stereomeric form.Example 4: Compound A inhibits in vitro growth of ovarian carcinoma cells

[0100] FIG. 9 shows the growth inhibition in human ovarian carcinoma cells (OVCAR3 cell line) in an in vitro model treated with Compound A or control (vehicle). Briefly, OVCAR3 human ovarian carcinoma cells were plated and incubated in vitro and treated with Compound A or control (vehicle). Cell density (confluence) was measured before and after drug treatment with Compound A (lOOnM). As shown in FIG. 9, all reductions in tumor growth in response to Compound A treatments reached significance with p values < 0.05 as determined by one-way ANOVA.Example 5: Compound A inhibits in vivo growth of melanoma cells

[0101] FIGS. 10 and 11 show the growth inhibition in vivo of melanoma cells by Compound A. LOX melanoma cells (FIG. 10) and MB4667 melanoma cells (FIG. 11) were used to establish tumors in c57 / B16 mice. Tumor size was measured by caliper and once the tumors reached approximately 100mm3in size, treatments were initiated with intraperitoneal injection of Compound A (lOOnM) three times weekly. As shown in FIGS. 10 and 11, all reductions in tumor growth in response to Compound A treatments reached significance with p values < 0.05 as determined by one-way ANOVA with Fisher’s LSD post test.

[0102] The various features and processes described above may be used independently of one another or may be combined in various ways. All possible combinations and subcombinations are intended to fall within the scope of this disclosure. In addition, certain method or process blocks may be omitted in some implementations. The example systems and components described herein may be configured differently than described. For example, elements may be added to, removed from, or rearranged compared to the disclosed example embodiments.

[0103] Conditional language used herein, such as, among others, “can,” “could,” “may,” “e.g.,” and the like, unless specifically stated otherwise, or otherwise understood withinthe context as used, is generally intended to convey that certain embodiments include, while other embodiments do not include, certain features, elements, and / or steps. Thus, such conditional language is not generally intended to imply that features, elements and / or steps are in any way required for one or more embodiments or that one or more embodiments necessarily include logic for deciding, with or without author input or prompting, whether these features, elements and / or steps are included or are to be performed in any particular embodiment. The terms “comprising,” “including,” “having,” and the like are synonymous and are used inclusively, in an open-ended fashion, and do not exclude additional elements, features, acts, operations, and so forth. Also, the term “or” is used in its inclusive sense (and not in its exclusive sense) so that when used, for example, to connect a list of elements, the term “or” means one, some, or all of the elements in the list. Throughout this description all ranges described include all values and sub-ranges therein, unless otherwise specified. Additionally, the indefinite article “a” or “an” carries the meaning of “one or more” throughout the description, unless otherwise specified.

[0104] Whereas certain example embodiments have been described, these embodiments have been presented by way of example only and are not intended to limit the scope of the inventions disclosed herein. Thus, nothing in the foregoing description is intended to imply that any particular feature, characteristic, step, or module is necessary or indispensable. Indeed, the novel methods and systems described herein may be embodied in a variety of other forms; furthermore, various omissions, substitutions, and changes in the form of the methods and systems described herein may be made without departing from the spirit of the inventions disclosed herein. The accompanying claims and their equivalents are intended to cover such forms or modifications as would fall within the scope and spirit of certain of the inventions disclosed herein.

Claims

CLAIMSWhat is claimed is:

1. A compound having the chemical structure:X2is H or OH; and, X3is F, Cl, Br, or I; and pharmaceutically acceptable metabolites, polymorphs, salts, solvates, hydrates, or clathrates thereof.

2. The compound of claim 1, having the chemical structure:

3. The compound of claim 1, wherein X1is OH.

4. The compound of claim 1, wherein X1is H.

5. The compound of claim 1, wherein X1is OC(=O)CH2Br.

6. The compound of claim 1, wherein X2is OH.

7. The compound of claim 1, wherein X2is H.

8. The compound of claim 1, wherein X1is OH and X2is OH.

9. The compound of claim 1, wherein X1is H and X2is OH.

10. The compound of claim 1, wherein X1is -OC(=O)CH2Br and X2is OH.

11. The compound of claim 1, wherein X1is -OC(=O)CH2Br and X2is H.

12. The compound of claim 1, wherein X3is F.

13. The compound of claim 1, wherein X3is Cl.

14. The compound of claim 1, wherein X3is Br.

15. The compound of claim 1, wherein X3is I.

16. A pharmaceutical composition comprising at least one of the compounds of claim 1, and / or pharmaceutically acceptable metabolites, polymorphs, salts, solvates, hydrates, or clathrates thereof.

17. The pharmaceutical composition of claim 16, wherein said composition is formulated for intravenous, oral, subcutaneous, or intramuscular administration.

18. A method for treating cancer in a patient in need of such a treatment, said method comprising administering to the patient a therapeutically effective amount of a compound of any one of claims 1-15, or a pharmaceutical composition of any one of claims 16 or 17.

19. The method of claim 18, wherein the cancer is one of ovarian cancer, malignant melanoma, and triple-negative breast cancer.

20. The method of claims 18 or 19, further comprising administering an oncolytic agent to the patient.

21. The method of claim 20, wherein the oncolytic agent is at least one oncolytic agent selected from aldesleukin, binimetinib, encorafenib, cobimetinib, dabrafenib mesylate, dacarbazine, doxorubicin, encorafenib, aldesleukin (IL-2), paclitaxel, talimogene laherparepvec, recombinant interferon a-2b, peginterferon a-2b, ipilimumab, pembrolizumab, trametinib, binimetinib, nivolumab, vemurafenib, tebentafusp, or combinations thereof.

22. The method of any one of claims 18-21, further comprising administering an immunotherapy to the patient.

23. The method of claim 22, wherein the immunotherapy comprises a PD-1 inhibitor or a PD-L1 inhibitor.

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