Treatment of cancer using combinations of an EED inhibitor and an AKT inhibitor

Abstract

The present disclosure relates to methods of treating cancer in a subject, comprising administering to the subject (a) Compound 1, or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor. In some embodiments, the AKT inhibitor is capivasertib. In some embodiments, the cancer in the subject is breast cancer or prostate cancer

Description

WSGR Ref. 47134-779.601TREATMENT OF CANCER USING COMBINATIONS OF AN EED INHIBITOR AND AN AKTINHIBITORCROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 703,090 filed October 3, 2024; which is hereby incorporated by reference in its entirety.BACKGROUND OF THE INVENTION

[0002] Cancer is the leading cause of death worldwide and the second leading cause in the United States. Prostate cancer is the most common cancer among men in the United States, with about one in nine men diagnosed in their lifetime. Many cancers, including prostate cancer, remain one of the leading causes of death and new treatments are needed. Poly comb repressive complex 2 (PRC2) dysregulation is linked to poor prognosis in cancers because the PRC2-controlled epigenetic chromatin silencing mechanism plays a key role in regulating cellular functions such as cell growth and differentiation. The dysregulation of PRC2 activity can lead to tumorigenesis in a wide range of cancers including prostate cancer, breast cancer, and hematological malignancies. In addition, misregulation of PRC2 function in prostate cancer contributes not only to tumorigenesis, but also therapeutic resistance. Mutations and alterations in the PI3K / AKT / PTEN pathway, another crucial cell growth, survival, metabolism, and proliferation pathway, are also common in cancers, including prostate cancer. For example, approximately 40% of prostate cancer tumors harbor a form of PI3K pathway alteration, with PTEN loss being the most frequent, followed by PIK3CA and AKT1 mutations. These alterations are critical because they are associated with more aggressive tumor behavior, resistance to standard therapies (like androgen deprivation therapy in the case of prostate cancer), and may serve as biomarkers for targeted therapies, such as AKT inhibitors. Given the need for new treatments in the treatment of cancer and the critical role of both PRC2 function and PI3K / AKT / PTEN pathway signaling in cancer, inhibition of both pathways in subjects with PI3K / AKT / PTEN pathway alterations may provide a new opportunity for the treatment of cancers in the subjects.

[0003] The present disclosure relates to methods of treating cancer in a subject, comprising administering to the subject (a) an embryonic ectoderm development (EED) inhibitor, and (b) an AKT inhibitor. In some embodiments, the EED inhibitor is Compound 1, or a pharmaceutically acceptable salt thereof.WSGR Ref. 47134-779.601SUMMARY OF THE INVENTION

[0004] Provided herein are methods of treating cancer in a subject, comprising administering to the subject (a) Compound(Compound 1), or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor.

[0005] Also provided herein are methods of treating cancer in a subject, wherein the cancer in the subject has been determined to comprise one or more PIK3CA / AKT1 / PTEN -alterations, comprising administering to the subject (a) Compound 1, or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor. In some embodiments, the cancer is breast cancer. In other embodiments, the cancer is prostate cancer.INCORPORATION BY REFERENCE

[0006] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.DETAILED DESCRIPTION OF THE INVENTION

[0007] As used in the specification and appended claims, unless specified to the contrary, the following terms have the meaning indicated below.

[0008] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “an agent” includes a plurality of such agents, and reference to “the cell” includes reference to one or more cells (or to a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formulae, all combinations and sub -combinations of ranges and specific embodiments therein are intended to be included. The term “about” when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range, in some instances, will vary between 1% and 15% of the stated number or numerical range. The term “comprising” (and related terms such as “comprise” or “comprises” or “having” or “including”) is not intended to exclude that in other certain embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, described herein, “consist of’ or “consist essentially of’ the described features.WSGR Ref. 47134-779.601

[0009] “Administering” when used in conjunction with a therapeutic, including androgen receptor inhibitors and embryonic ectoderm development (EED) inhibitors, means to administer a therapeutic systemically or locally, as directly into or onto a target tissue, or to administer a therapeutic to a subject whereby the therapeutic positively impacts the tissue to which it is targeted. Thus, as used herein, the term “administering”, when used in conjunction with a composition described herein, can include, but is not limited to, providing a composition into or onto the target tissue; providing a composition systemically to a subject by, e.g., oral administration whereby the therapeutic reaches the target tissue or cells. “Administering” a composition may be accomplished by injection, topical administration, and oral administration or by other methods alone or in combination with other known techniques.

[0010] The term “AKT inhibitor,” as used herein, means an inhibitor of the serine / threonine protein kinase Akt (protein kinase B), which may result in inhibition of the PI3K / Akt signaling pathway in cells, including cells in subject, such as cells comprising a cancer in a subject. In it specifically contemplated herein that the AKT inhibitor utilized in the methods described herein may be an inhibitor of one or more of the 3 isoforms of AKT, including AKT1, AKT2, and AKT3. In some embodiments, the AKT inhibitor is an inhibitor of all of AKT1, AKT2, and AKT3. In other embodiments, the AKT inhibitor is selective inhibitor of one of AKT1, AKT2, and AKT3.

[0011] The terms “determine,” “determined,” and “determining,” and the like, as used herein mean that it has been established that a pre-condition in a subject exists, or a condition precedent with respect to a subject has been satisfied, prior to the administration to the subject of an embryonic ectoderm development (EED) inhibitors and an AKT inhibitor. AKT inhibitors which may be used according to the methods disclosed herein include, but are not limited to, ipatasertib (GDC-0068), capivasertib (AZD5363), MK2206, afiiresertib (GSK2110183), uprosertib (GSK2141795), perifosine (KRX- 0401), PHT-427 (CS-0223), and Akti-1 / 2. In some embodiments, the AKT inhibitor is ipatasertib (GDC-0068). In some embodiments, the AKT inhibitor is capivasertib (AZD5363). In some embodiments, the AKT inhibitor is MK2206. In some embodiments, the AKT inhibitor is afiiresertib (GSK2110183). In some embodiments, the AKT inhibitor is uprosertib (GSK2141795). In some embodiments, the AKT inhibitor is perifosine (KRX- 0401). In some embodiments, the AKT inhibitor is PHT-427 (CS-0223). In some embodiments, the AKT inhibitor is Akti-1 / 2.

[0012] The term “embryonic ectoderm development (EED) inhibitor,” as used herein, means an agent that inhibits the function of the embryonic ectoderm development (EED) protein, including by binding to the protein.

[0013] The term “pharmaceutically acceptable”, as used herein, means a carrier, diluent or excipient that is compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0014] The term “pharmaceutical composition” means a composition comprising at least one active ingredient, whereby the composition is amenable to investigation for a specified, efficacious outcome in a mammal (for example, without limitation, a human). Those of ordinary skill in the art will understand andWSGR Ref. 47134-779.601 appreciate the techniques appropriate for determining whether an active ingredient has a desired efficacious outcome based upon the needs of the artisan.

[0015] The term “one or more PIK3CA / AKT1 / PTEN alterations,” as used herein, means one or more alterations in one or more of the PIK3CA, AKT1, and / or PTEN proteins in a subject having cancer, such as prostate cancer, compared to the PIK3CA, AKT1, and / or PTEN proteins in a subject not having cancer. Such alterations in a subject having cancer may include, but are not limited to, expression of protein having one or more mutations in the amino acid sequence of the protein, over-expression of the protein, or decreased expression of the protein. For example, the PIK3CA protein may be overexpressed in a subject having cancer, or there may be mutations in the amino acid sequence of the protein, for example E542K, E545K, and H1047R, that contribute to the development of cancer in the subject. The AKT1 protein may be overexpressed in a subject having cancer, or there may be mutations in the amino acid sequence of the protein, for example E17K, that contribute to the development of cancer in the subject. The PTEN protein may not be expressed to the same degree in a subject having cancer compared to a subject not having cancer (due to, e.g., gene deletions or epigenetic silencing) or there may be mutations in the amino acid sequence of the protein, for example E17K, that contribute to the development of cancer in the subject. Such alterations in the PIK3CA, AKT1, and / or PTEN proteins in a subject having cancer may be the result of alterations in the genes that lead to expression of the proteins, including amplification of the gene, gene deletions, or missense or non-sense mutations in the gene sequence, or the result of epigenetic silencing of the genes.

[0016] The term “PTEN null,” as used herein, means the loss or absence of the PTEN (Phosphatase and Tensin Homolog) gene function in a cell, which may be caused by deletion of the gene, mutation of the gene, or hypermethylation in the promoter region of the gene resulting in the loss of cellular PTEN expression and / or function.

[0017] As used herein, the term “therapeutic” means an agent utilized to treat, combat, ameliorate, prevent, or improve an unwanted condition or disease of a subject.

[0018] A “therapeutically effective amount” or “effective amount” as used herein refers to the amount of active compound or pharmaceutical agent that elicits a biological or medicinal response in a tissue, system, animal, individual or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes one or more of the following: (1) preventing the disease; for example, preventing a disease, condition or disorder in an individual that may be predisposed to the disease, condition or disorder but does not yet experience or display the pathology or symptomatology of the disease, (2) inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and / or symptomatology), and (3) ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and / or symptomatology).WSGR Ref. 47134-779.601

[0019] The terms “treat,” “treated,” “treatment,” or “treating” as used herein refers to both therapeutic treatment in some embodiments and prophylactic or preventative measures in other embodiments, wherein the object is to prevent or slow (lessen) an undesired physiological condition, disorder, or disease, or to obtain beneficial or desired clinical results. For the purposes described herein, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total), whether detectable or undetectable, or enhancement or improvement of the condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment. A prophylactic benefit of treatment includes prevention of a condition, retarding the progress of a condition, stabilization of a condition, or decreasing the likelihood of occurrence of a condition.

[0020] Provided herein are methods of treating cancer in a subject, comprising administering to the subject (a) Compound(Compound 1), or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor.

[0021] In some embodiments are provided methods of treating cancer in a subject, wherein the cancer in the subject has been determined to comprise one or more PIK3CA / AKTl / PTEN-alterations, comprising administering to the subject (a) Compound 1, or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor. In some embodiments, the cancer in the subject has been determined to comprise one or more PIK3CA alterations. In some embodiments, the cancer in the subject has been determined to comprise one or more AKT1 alterations. In some embodiments, the cancer in the subject has been determined to comprise one or more PTEN alterations. In some embodiments, the cancer in the subject has been determined to be PTEN-null. In some embodiments, cancer in the subject has been determined to comprise one or more PI3KA / AKT1 / PTEN alterations as detected by an FDA-approved test. In some embodiments, the cancer in the subject has been determined to comprise one or more one or more PI3KA alterations as detected by an FDA-approved test. In other embodiments, the cancer in the subject has been determined to comprise one or more one or more AKT1 alterations as detected by an FDA -approved test. In other embodiments, the cancer in the subject has been determined to comprise one or more one or more PTEN alterations as detected by an FDA -approved test. In other embodiments, the cancer in the subject has been determined to be PTEN-null as detected by an FDA -approved test.WSGR Ref. 47134-779.601

[0022] In some embodiments of the methods disclosed herein, the subject has received prior administration of one or more chemotherapeutic agents prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor. In some embodiments, the subject has received up to one chemotherapeutic treatments prior the administration to the subject prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

[0023] In some embodiments of the methods disclosed herein, the subject has not been administered a chemotherapeutic agent prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

[0024] In some embodiments of the methods disclosed herein, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject sequentially or simultaneously. In other embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject sequentially. In still further embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject on the same day. In still further embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject within a 24-hour period. In another embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject within a 12-hour period, or a 10-hour period, or an 8-hour period, or a 6-hour period, or a 4-hour period, or a 2-hour period, or within an hour of each of other. In another embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject simultaneously. In a further embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject once per day or twice per day. In another embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject once per day. In another embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject twice per day. In a further embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to the subject once per day and the AKT inhibitor is administered to the subject twice per day.

[0025] Also disclosed herein are any of the methods of treating cancer disclosed herein, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject with food or without food. In an embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject with food.

[0026] In some embodiments of the methods disclosed herein, the subject has been administered at least one endocrine-based therapy prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor. In some embodiments, the prior endocrine therapy comprises a selective estrogen receptor degrader (SERD), a selective estrogen receptor modulator (SERM), a selective estrogen receptor covalent antagonist (SERCA), a selective human estrogen receptor agonist (ShERPA), an aromatase inhibitor (Al), or a combination thereof. In someWSGR Ref. 47134-779.601 embodiments of the methods disclosed herein, the cancer in the subject has progressed on the at least one endocrine -based therapy prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor. In some embodiments, the prior endocrine therapy comprises a selective estrogen receptor degrader (SERD), a selective estrogen receptor modulator (SERM), a selective estrogen receptor covalent antagonist (SERCA), a selective human estrogen receptor agonist (ShERPA), an aromatase inhibitor (Al), or a combination thereof.

[0027] Also provided herein are methods of treating cancer in a subject disclosed herein, comprising administering to the subject a therapeutically effective amount of (a) Compound(Compound 1), or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor, wherein Compound 1 is in crystalline form. Also provided herein are such methods wherein the crystalline form of Compound 1 is an anhydrous form. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 8. 1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 9.6° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.7° ± 0.2° 2-theta, 19.7° ± 0.2° 2-theta, and 22.0° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 9.8° ± 0.2° 2-theta, 15.2° ± 0.2° 2-theta, and 17.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of about 172 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 205 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C, or from about 207 °C to about 210 °C, or from about 208 °C to about 210 °C, or from about 209 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 1% upon heating the sample from about 25 °C to a temperature prior to melting. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 1% upon heating the sample from about 25 °C to about 380 °C.

[0028] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta. Also provided herein are suchWSGR Ref. 47134-779.601 methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta and 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ± 0.2° 2- theta, and 24. 1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 15.4° ± 0.2° 2-theta, 21.0° ± 0.2° 2-theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0029] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further a peak in an x-ray powder diffraction (XRPD) pattern at 15.4° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further a peak in an x-ray powder diffraction (XRPD) pattern at 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ± 0.2° 2-theta, and 24.1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 21.0° ± 0.2° 2-theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein the crystalline form of Compound 1WSGR Ref. 47134-779.601 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0030] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits peaks in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta and 15.4° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x- ray powder diffraction (XRPD) pattern at 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ± 0.2° 2-theta, and 24. 1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 21.0° ± 0.2° 2- theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0031] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits peaks in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta and 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 15.4° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ± 0.2° 2-theta, and 24. 1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 21.0° ± 0.2° 2- theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C. Also providedWSGR Ref. 47134-779.601 herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0032] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 10% degradation when stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days.

[0033] Also provided herein are such methods wherein the crystalline form of Compound 1 (a) exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 8.1° ± 0.2° 2-theta, and (b) exhibits less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 (a) exhibits peaks in an x-ray powder diffraction (XRPD) pattern at 9.6° ± 0.2° 2-theta, 5.7° ± 0.2° 2-theta, 19.7° ± 0.2° 2-theta, and 22.0° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days.

[0034] Also provided herein are such methods wherein the crystalline form of Compound 1 (a) exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta, and (b) exhibits less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) peaks in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta, 13.7° ± 0.2° 2-theta, and 19.2° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days.

[0035] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 10% degradation when the crystalline forms are stored at 40 °C and 75% relative humidity forWSGR Ref. 47134-779.601 at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days.

[0036] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) a peak in an x-ray powder diffraction (XRPD) pattern at 8.1° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) peaks in an x-ray powder diffraction (XRPD) pattern at 9.6° ± 0.2° 2-theta, 5.7° ± 0.2° 2-theta, 19.7° ± 0.2° 2-theta, and 22.0° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days.

[0037] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) peaks in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta, 13.7° ± 0.2° 2-theta, and 19.2° ± 0.2° 2- theta, and (b) less than about 10% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days.

[0038] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 10% degradation when the crystalline form is stored at 60 °C for at least one week. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 60 °C for at least one week.

[0039] In some embodiments of the methods disclosed herein, the AKT inhibitor is selected from ipatasertib (GDC-0068), capivasertib (AZD5363), MK2206, afuresertib (GSK2110183), uprosertib (GSK2141795), perifosine (KRX- 0401), PHT-427 (CS-0223), and Akti-1 / 2. In some embodiments, the AKT inhibitor is ipatasertib (GDC-0068). In some embodiments, the AKT inhibitor is capivasertib (AZD5363). In some embodiments, the AKT inhibitor is MK2206. In some embodiments, the AKTWSGR Ref. 47134-779.601 inhibitor is afuresertib (GSK2110183). In some embodiments, the AKT inhibitor is uprosertib (GSK2141795). In some embodiments, the AKT inhibitor is perifosine (KRX- 0401). In some embodiments, the AKT inhibitor is PHT-427 (CS-0223). In some embodiments, the AKT inhibitor is Akti-1 / 2.

[0040] In some of embodiments of the methods disclosed herein, the cancer in the subject is selected from diffused large B cell lymphoma (DLBCL), follicular lymphoma, other lymphomas, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, cancer, breast cancer, bile duct and gallbladder cancers, bladder carcinoma, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal carcinoma, ovarian cancer, pancreatic cancer, renal cell carcinoma, prostate cancer, rectal cancer, thyroid cancers, parathyroid tumors, uterine tumors, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), Kaposi sarcoma, synovial sarcoma, osteosarcoma and Ewing's sarcoma.

[0041] In some of embodiments of the methods disclosed herein, the cancer in the subject is breast cancer. In some embodiments, the breast cancer in the subject is selected from HR-positive breast carcinoma, HR-negative breast carcinoma, primary breast ductal carcinoma, mammary adenocarcinoma, HR-positive mammary ductal carcinoma, HR- negative mammary ductal carcinoma, HER2 -positive mammary ductal carcinoma, HER2 -positive breast cancer, luminal breast cancer, triple negative breast cancer (TNBC), ER-positive breast cancer, ER-negative breast cancer, PR-positive breast cancer, PR- negative breast cancer, HR-positive / HER-2 negative breast cancer, HR-negative / HER-2 negative breast cancer, HR-positive / HER-2 positive breast cancer, and HR-negative / HER-2 positive breast cancer, and unclassified breast cancer. In one embodiment, the breast cancer in the subject is HR-positive / HER-2 negative breast cancer. In another embodiment, the breast cancer in the subject is HR-positive / HER-2 negative, locally advanced or metastatic breast cancer. In another embodiment, the breast cancer in the subject is HR-positive / HER-2 negative, locally advanced or metastatic breast cancer, with one or more PIK3CA / AKT1 / PTEN -alterations. In another embodiment, the breast cancer in the subject is HR- positive / HER-2 negative, locally advanced or metastatic breast cancer, with one or more PIK3CA / AKT1 / PTEN -alterations as detected by an FDA-approved test. In another embodiment, the breast cancer in the subject is HR-positive / HER-2 negative, locally advanced or metastatic breast cancer, with one or more PIK3CA / AKT1 / PTEN -alterations as detected by an FDA -approved test following progression on at least one endocrine-based regimen in the metastatic setting or recurrence on or within 12 months of completing adjuvant therapy.

[0042] In some of embodiments of the methods disclosed herein, the cancer in the subject is triple negative breast cancer (TNBC). In some embodiments, the TNBC is selected from the group consisting of basal -like TNBC, mesenchymal TNBC, immunomodulatory TNBC, and luminal androgen receptor TNBC.WSGR Ref. 47134-779.601

[0043] In some of embodiments of the methods disclosed herein, the cancer in the subject is prostate cancer. In some embodiments, the prostate cancer in the subject is selected from metastatic prostate cancer, non-metastatic prostate cancer, metastatic castration-resistant prostate cancer, metastatic castration-sensitive prostate cancer, localized high risk prostate cancer, recurrent prostate cancer, non- metastatic castration-resistant prostate cancer, non-metastatic castration-sensitive prostate cancer, androgen receptor inhibitor-sensitive prostate cancer, androgen receptor inhibitor-resistant prostate cancer, androgen receptor-dependent prostate cancer, androgen receptor-independent prostate cancer, neuroendocrine prostate cancer (NEPC), metastatic neuroendocrine prostate cancer (NEPC), prostate cancer with small cell features, metastatic prostate cancer with small cell features, and aggressive-variant prostate cancer. In some embodiments, the prostate cancer in the subject is metastatic prostate cancer. In some embodiments, the prostate cancer in the subject is non -metastatic prostate cancer. In some embodiments, the prostate cancer in the subject is metastatic castration-resistant prostate cancer. In some embodiments, the prostate cancer in the subject is metastatic castration-sensitive prostate cancer. In some embodiments, the prostate cancer in the subject is localized high risk prostate cancer. In some embodiments, the prostate cancer in the subject is recurrent prostate cancer. In some embodiments, the prostate cancer in the subject is non-metastatic castration-resistant prostate cancer. In some embodiments, the prostate cancer in the subject is non -metastatic castration-sensitive prostate cancer. In some embodiments, the prostate cancer in the subject is androgen receptor inhibitor-sensitive prostate cancer. In some embodiments, the prostate cancer in the subject is androgen receptor inhibitor-resistant prostate cancer. In some embodiments, the prostate cancer in the subject is androgen receptor-dependent prostate cancer. In some embodiments, the prostate cancer in the subject is androgen receptor-independent prostate cancer. In some embodiments, the prostate cancer in the subject is neuroendocrine prostate cancer (NEPC). In some embodiments, the prostate cancer in the subject is metastatic neuroendocrine prostate cancer (NEPC). In some embodiments, the prostate cancer in the subject is prostate cancer with small cell features. In some embodiments, the prostate cancer in the subject is metastatic prostate cancer with small cell features. In some embodiments, the prostate cancer in the subject is aggressive-variant prostate cancer.

[0044] In some embodiments, the cancer in the subject is prostate cancer and the subject is not administered an androgen receptor inhibitor in combination with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the cancer in the subject is prostate cancer and the subject is not administered an androgen receptor inhibitor selected from apalutamide, darolutamide, enzalutamide, or abiraterone. In some embodiments, the cancer in the subject is prostate cancer and the subject is not administered apalutamide in combination with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the cancer in the subject is prostate cancer and the subject is not administered darolutamide in combination with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the cancer in the subject is prostate cancer and the subject is not administered enzalutamide in combination withWSGR Ref. 47134-779.601Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the cancer in the subject is prostate cancer and the subject is not administered abiraterone in combination with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor.

[0045] In other embodiments are provided the methods of treating cancer in a subject described herein, wherein the subject is administered a therapeutically effective amount of (a) Compound 1, or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor, and is further administered one or more additional therapeutic agents. In further embodiments are provided such methods, wherein the one or more additional therapeutic agents is selected from an androgen receptor degrader, a chemotherapeutic agent, a mitotic inhibitors, an antimetabolites, a platinum -based agents, histone deacetylase (HDAC) inhibitors, CD30-directed antibody-drug conjugates, famesyl transferase inhibitors, SYK inhibitors, JAK inhibitors, PI3K pathway inhibitors, immunomodulatory agents, radiopharmaceuticals, PARP inhibitors, or combinations thereof. In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from chemotherapeutic agents.

[0046] In other embodiments are provided such methods wherein the cancer in the subject is prostate cancer and the androgen receptor degrader is a PROteolytic-TArgeting-Chimera (PROTAC®) androgen receptor (AR) degrader. In further embodiments, the androgen receptor degrader is bavdegalutamide (ARV- 110).

[0047] In further embodiments are provided such methods, wherein the chemotherapeutic agents are selected from actinomycin, azacytidine, azathioprine, bendamustine, bleomycin, bortezomib, chlorambucil, cyclophosphamide, daunorubicin, doxifluridine, doxorubicin, epirubicin, epothilone, etoposide, idarubicin, irinotecan, lurbinectedin, mechlorethamine, mitoxantrone, teniposide, topotecan, valrubicin, vemurafenib, vinblastine, vincristine, and vindesine. In some embodiments, the chemotherapeutic agent is actinomycin. In some embodiments, the chemotherapeutic agent is azacytidine. In some embodiments, the chemotherapeutic agent is azathioprine. In some embodiments, the chemotherapeutic agent is bleomycin. In some embodiments, the chemotherapeutic agent is bortezomib. In some embodiments, the chemotherapeutic agent is chlorambucil. In some embodiments, the chemotherapeutic agent is cyclophosphamide. In some embodiments, the chemotherapeutic agent is daunorubicin. In some embodiments, the chemotherapeutic agent is doxifluridine. In some embodiments, the chemotherapeutic agent is doxorubicin. In some embodiments, the chemotherapeutic agent is epirubicin. In some embodiments, the chemotherapeutic agent is epothilone. In some embodiments, the chemotherapeutic agent is etoposide. In some embodiments, the chemotherapeutic agent is idarubicin. In some embodiments, the chemotherapeutic agent is irinotecan. In some embodiments, the chemotherapeutic agent is lurbinectedin. In some embodiments, the chemotherapeutic agent is mechlorethamine. In some embodiments, the chemotherapeutic agent is mitoxantrone. In some embodiments, the chemotherapeutic agent is teniposide. In some embodiments, the chemotherapeutic agent is topotecan. In some embodiments, the chemotherapeutic agent is valrubicin. In some embodiments, the chemotherapeutic agent is vemurafenib. In some embodiments, the chemotherapeuticWSGR Ref. 47134-779.601 agent is vinblastine. In some embodiments, the chemotherapeutic agent is vincristine. In some embodiments, the chemotherapeutic agent is vindesine.

[0048] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from mitotic inhibitors. In further embodiments are provided such methods, wherein the mitotic inhibitors are selected from paclitaxel, docetaxel, cabazitaxel, tesetaxel, and nab-paclitaxel. In some embodiments, the mitotic inhibitor is paclitaxel. In some embodiments, the mitotic inhibitor is docetaxel. In some embodiments, the mitotic inhibitor is cabazitaxel. In some embodiments, the mitotic inhibitor is tesetaxel. In some embodiments, the mitotic inhibitor is nab- paclitaxel.

[0049] In yet other embodiments, the one or more additional therapeutic agents are selected from antimetabolites. In some embodiments, the one or more antimetabolites are selected from azacytidine, 6- mercaptopurine, capecitabine, hydroxyurea, cladribine, pralatrexate, thioguanine, decitabine, clofarabine, nelarabine, fludarabine, 5 -fluorouracil, gemcitabine, cytarabine, pemetrexed, and methotrexate, cytarabine (Ara-C), floxuridine, fludarabine, pentostatin, and trifluridine / tipiracil combination. In some embodiments, the one or more antimetabolites are selected from 6-mercaptopurine, capecitabine, hydroxyurea, cladribine, pralatrexate, thioguanine, decitabine, clofarabine, nelarabine, fludarabine, 5- fluorouracil, gemcitabine, cytarabine, pemetrexed, and methotrexate.

[0050] In some embodiments, the antimetabolite is azacytidine. In some embodiments, the antimetabolite is 6-mercaptopurine. In some embodiments, the antimetabolite is capecitabine. In some embodiments, the antimetabolite is hydroxyurea. In some embodiments, the antimetabolite is cladribine. In some embodiments, the antimetabolite is pralatrexate. In some embodiments, the antimetabolite is thioguanine. In some embodiments, the antimetabolite is decitabine. In some embodiments, the antimetabolite is clofarabine. In some embodiments, the antimetabolite is nelarabine. In some embodiments, the antimetabolite is fludarabine. In some embodiments, the antimetabolite is 5 -fluorouracil. In some embodiments, the antimetabolite is gemcitabine. In some embodiments, the antimetabolite is cytarabine. In some embodiments, the antimetabolite is pemetrexed. In some embodiments, the antimetabolite is methotrexate. In some embodiments, the antimetabolite is cytarabine (Ara-C). In some embodiments, the antimetabolite is floxuridine. In some embodiments, the antimetabolite is fludarabine. In some embodiments, the antimetabolite is pentostatin. In some embodiments, the antimetabolite is a trifluridine / tipiracil combination.

[0051] In some embodiments, the one or more additional therapeutic agents are selected from platinumbased agents. In some embodiments, the platinum -based agents are selected from cisplatin, carboplatin, oxaliplatin, nedaplatin, lobaplatin, triplatin tetranitrate, pheanthriplatin, picoplatin, and satraplatin. In further embodiments, the platinum -based agent is cisplatin. In further embodiments, the platinum -based agent is carboplatin. In further embodiments, the platinum-based agent is oxaliplatin. In further embodiments, the platinum-based agent is nedaplatin. In further embodiments, the platinum-based agent is lobaplatin. In further embodiments, the platinum -based agent is triplatin tetranitrate. In furtherWSGR Ref. 47134-779.601 embodiments, the platinum-based agent is pheanthriplatin. In further embodiments, the platinum -based agent is picoplatin. In further embodiments, the platinum-based agent is satraplatin.

[0052] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from histone deacetylase (HDAC) inhibitors. In further embodiments are provided such methods, wherein the histone deacetylase (HDAC) inhibitors are selected from vorinostat, romidepsin, belinostat, tucidinostat, panobinostat, mocetinostat, givinostat, resmiostat, abexinostat, ricolinostat, entinostat, tinostamustin, fimepinostat, CXD-101, quisinostat, and chidamide. In further embodiments, the histone deacetylase (HDAC) inhibitor is vorinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is romidepsin. In further embodiments, the histone deacetylase (HDAC) inhibitor is belinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is tucidinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is Panobinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is mocetinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is givinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is resmiostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is abexinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is ricolinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is entinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is tinostamustin. In further embodiments, the histone deacetylase (HDAC) inhibitor is fimepinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is CXD-101. In further embodiments, the histone deacetylase (HDAC) inhibitor is quisinostat. In further embodiments, the histone deacetylase (HDAC) inhibitor is chidamide.

[0053] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from CD30-directed antibody-drug conjugates. In further embodiments are provided such methods, wherein the CD30-directed antibody-drug conjugates are selected from brentuximab vedotin and SGN-CD30C. In further embodiments, the CD30-directed antibody-drug conjugate is brentuximab vedotin. In further embodiments, the CD30-directed antibody-drug conjugate is SGN-CD30C.

[0054] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from famesyl transferase inhibitors. In further embodiments are provided such methods, wherein the famesyl transferase inhibitors are selected from antroquinonol, BMS-214662, L778123, L744832, FTI-276, FTI-277, manumycin A, LB-42708, moverastin, PDI6954I, ABT-100, FTI- 2153, tipifamib and lonafamib. In further embodiments, the famesyl transferase inhibitor is antroquinonol. In further embodiments, the famesyl transferase inhibitor is BMS-214662. In further embodiments, the famesyl transferase inhibitor is L778123. In further embodiments, the famesyl transferase inhibitor is L744832. In further embodiments, the famesyl transferase inhibitor is FTI-276. In further embodiments, the famesyl transferase inhibitor is FTI-277. In further embodiments, the famesyl transferase inhibitor is manumycin A. In further embodiments, the famesyl transferase inhibitor is LB-42708. In further embodiments, the famesyl transferase inhibitor is moverastin. In further embodiments, the famesylWSGR Ref. 47134-779.601 transferase inhibitor is PD169541. In further embodiments, the famesyl transferase inhibitor is ABT-100. In further embodiments, the famesyl transferase inhibitor is FTI-2153. In further embodiments, the famesyl transferase inhibitor is tipifamib. In further embodiments, the famesyl transferase inhibitor is lonafamib.

[0055] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from SYK inhibitors. In further embodiments are provided such methods, wherein the SYK inhibitors are selected from fostamatinib (R788), entospletinib (GS-9973), cerdulatinib (PRT062070), and TAK-659. In further embodiments, the SYK inhibitor is fostamatinib (R788). In further embodiments, the SYK inhibitor is entospletinib (GS-9973). In further embodiments, the SYK inhibitor is cerdulatinib (PRT062070). In further embodiments, the SYK inhibitor is TAK-659.

[0056] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from JAK inhibitors. In further embodiments are provided such methods, wherein the JAK inhibitors are selected from tofacitinib, baricitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, and ruxolitinib. In further embodiments, the JAK inhibitor is tofacitinib. In further embodiments, the JAK inhibitor is baricitinib. In further embodiments, the JAK inhibitor is ruxolitinib. In further embodiments, the JAK inhibitor is upadacitinib. In further embodiments, the JAK inhibitor is fedratinib. In further embodiments, the JAK inhibitor is abrocitinib. In further embodiments, the JAK inhibitor is ruxolitinib.

[0057] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from PI3K pathway inhibitors. In further embodiments are provided such methods, wherein the PI3K pathway inhibitors are selected from taselisib (GDC-0032), GDC-0077, perifosine, idelalisib, buparlisib (BKM120), duvelisib, (IPI-145), copanlisib (BAY 80-6946), PX-866, dactolisib, CUDC-907, voxtalisib (SAR245409, XL765), ME-401, IPI-549, SF1126, RP6530, INK1117, pictilisib (GDC-0941), XL147 (SAR245408), palomid 529, GSK1059615, ZSTK474, and PWT33597. In further embodiments, the PI3K pathway inhibitor is taselisib (GDC-0032). In further embodiments, the PI3K pathway inhibitor is GDC-0077. In further embodiments, the PI3K pathway inhibitor is perifosine. In further embodiments, the PI3K pathway inhibitor is idelalisib. In further embodiments, the PI3K pathway inhibitor is buparlisib (BKM120). In further embodiments, the PI3K pathway inhibitor is duvelisib. In further embodiments, the PI3K pathway inhibitor is (IPI-145). In further embodiments, the PI3K pathway inhibitor is copanlisib (BAY 80-6946). In further embodiments, the PI3K pathway inhibitor is PX-866. In further embodiments, the PI3K pathway inhibitor is dactolisib. In further embodiments, the PI3K pathway inhibitor is CUDC-907. In further embodiments, the PI3K pathway inhibitor is voxtalisib (SAR245409, XL765). In further embodiments, the PI3K pathway inhibitor is ME- 401. In further embodiments, the PI3K pathway inhibitor is IPI-549. In further embodiments, the PI3K pathway inhibitor is SF1126. In further embodiments, the PI3K pathway inhibitor is RP6530. In further embodiments, the PI3K pathway inhibitor is INK1117. In further embodiments, the PI3K pathway inhibitor is pictilisib (GDC-0941). In further embodiments, the PI3K pathway inhibitor is XL147WSGR Ref. 47134-779.601(SAR245408). In further embodiments, the PI3K pathway inhibitor is palomid 529. In further embodiments, the PI3K pathway inhibitor is GSK1059615. In further embodiments, the PI3K pathway inhibitor is ZSTK474. In further embodiments, the PI3K pathway inhibitor is PWT33597.

[0058] In further embodiments are provided such methods, wherein the one or more additional therapeutic agents are selected from immunomodulatory agents. In further embodiments are provided such methods, wherein the immunomodulatory agents are selected from lenalidomide, PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 agents, T-cell immunoglobulin and ITIM domain (TIGIT) agents, TIM-3 inhibitors, and LAG-3 inhibitors.

[0059] In other embodiments are provided the methods disclosed herein, wherein the one or more additional therapeutic agents are selected from PD-1 inhibitors. In some embodiments, the PD-1 inhibitors are selected from pembrolizumab, nivolumab, cemiplimab, JTX-4014, spartalizumab (PDR001), camrelizumab (SHR1210), sintilimab (IBI308), tislelizumab (BGB-A317), toripalimab (JS 001), dostarlimab (TSR-042, WBP-285), INCMGA00012 (MGA012), AMP-224, and AMP-514 (MEDI0680).

[0060] In some embodiments, the PD-1 inhibitor is pembrolizumab. In some embodiments, the PD-1 inhibitor is nivolumab. In some embodiments, the PD-1 inhibitor is cemiplimab. In some embodiments, the PD-1 inhibitor is spartalizumab (PDR001). In some embodiments, the PD-1 inhibitor is camrelizumab (SHR1210). In some embodiments, the PD-1 inhibitor is sintilimab (IBI308). In some embodiments, the PD-1 inhibitor is tislelizumab (BGB-A317). In some embodiments, the PD-1 inhibitor is toripalimab (JS 001). In some embodiments, the PD-1 inhibitor is dostarlimab (TSR-042, WBP-285). In some embodiments, the PD-1 inhibitor is INCMGA00012 (MGA012). In some embodiments, the PD-1 inhibitor is AMP -224. In some embodiments, the PD-1 inhibitor is AMP-514 (MEDI0680).

[0061] In other embodiments are provided the methods disclosed herein, wherein the one or more additional therapeutic agents are selected from PD-L1 inhibitors. In some embodiments, the PD-L1 inhibitors are selected from atezolizumab, avelumab, durvalumab, MPDL3280A (RG7446), MDX-1105 (BMS-936559), BMS-935559, MSB0010718C, and MEDI4736.

[0062] In some embodiments, the PD-L1 inhibitors are selected from atezolizumab, avelumab, durvalumab, KN035, CK-301, AUNP12, CA-170, and BMS-986189.

[0063] In some embodiments, the PD-L1 inhibitor is atezolizumab. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, the PD-L1 inhibitor is durvalumab. In some embodiments, the PD-L1 inhibitor is MPDL3280A (RG7446). In some embodiments, the PD-L1 inhibitor is MDX-1105 (BMS-936559). In some embodiments, the PD-L1 inhibitor is BMS-935559. In some embodiments, the PD-L1 inhibitor is MSB0010718C. In some embodiments, the PD-L1 inhibitor is MEDI4736.

[0064] In further embodiments are provided such methods, wherein the CTLA-4 inhibitors are selected from ipilimumab and tremelimumab. In further embodiments, the CTLA-4 inhibitor is ipilimumab. In further embodiments, the CTLA-4 inhibitor is tremelimumab.

[0065] In further embodiments are provided such methods, wherein the T-cell immunoglobulin and ITIM domain (TIGIT) agents are selected from BMS-986207, BGB-A1217, tiragolumab, AB154,WSGR Ref. 47134-779.601ASP8374, MK-7684, CD112RCOM701, and LY3435151. In further embodiments, the TIGIT agent is BMS-986207. In further embodiments, the TIGIT agent is BGB-A1217. In further embodiments, the TIGIT agent is tiragolumab. In further embodiments, the TIGIT agent is AB154. In further embodiments, the TIGIT agent is ASP8374. In further embodiments, the TIGIT agent is MK-7684. In further embodiments, the TIGIT agent is CD112RCOM701. In further embodiments, the TIGIT agent is LY3435151.

[0066] In further embodiments are provided such methods, wherein the TIM-3 inhibitors are selected from Sym023, INCAGN02390, LY331367, Sym021, MBG453, BGB-A425, TSR-022, RO7121661, and LU3415244. In further embodiments, the TIM-3 inhibitor is Sym023. In further embodiments, the TIM-3 inhibitor is INCAGN02390. In further embodiments, the TIM-3 inhibitor is LY331367. In further embodiments, the TIM-3 inhibitor is Sym021. In further embodiments, the TIM-3 inhibitor is MBG453. In further embodiments, the TIM-3 inhibitor is BGB-A425. In further embodiments, the TIM-3 inhibitor is TSR-022. In further embodiments, the TIM-3 inhibitor is RO7121661. In further embodiments, the TIM-3 inhibitor is LU3415244.

[0067] In further embodiments are provided such methods, wherein the LAG-3 inhibitors are selected from relatlimab, tebotelimab, chlorogenic acid, RO-7247669, favezelimab, INCAGN-2385, IBI-110, eftilagimod alpha, Sym-022, LBL-007, ABL-501, HLX 26, IBI-323, ieramilimab, FS 118, EMB-02, and fmalimab. In further embodiments, the LAG-3 inhibitor is relatlimab. In further embodiments, the LAG-3 inhibitor is tebotelimab. In further embodiments, the LAG-3 inhibitor is chlorogenic acid. In further embodiments, the LAG-3 inhibitor is RO-7247669. In further embodiments, the LAG-3 inhibitor is favezelimab. In further embodiments, the LAG-3 inhibitor is INCAGN-2385. In further embodiments, the LAG-3 inhibitor is IBI-110,=. In further embodiments, the LAG-3 inhibitor is eftilagimod alpha. In further embodiments, the LAG-3 inhibitor is Sym-022. In further embodiments, the LAG-3 inhibitor is LBL-007. In further embodiments, the LAG-3 inhibitor is ABL-501. In further embodiments, the LAG-3 inhibitor is HLX 26,=. In further embodiments, the LAG-3 inhibitor is IBI-323. In further embodiments, the LAG-3 inhibitor is ieramilimab. In further embodiments, the LAG-3 inhibitor is FS 118. In further embodiments, the LAG-3 inhibitor is EMB-02. In further embodiments, the LAG-3 inhibitor is fmalimab.

[0068] In further embodiments are provided such methods, wherein the radiopharmaceuticals are selected from radio-ligand therapy agents. In some embodiments, the radiopharmaceutical is lutetium Lu 177 vipivotide tetraxetan.

[0069] In further embodiments are provided such methods, wherein the PARP inhibitors are selected from olaparib, rucaparib, niraparib, veliparib, fuzolaparib, CEP 9722, E7016, talazoparib, veliparib, pamiparib, AZD5305, AZD5135, AZD9574, IMP1734, DM5167, KU-0059436 (AZD2281), NMS-293, SNV-001, compounds disclosed in WO 2022 / 225934, compounds disclosed in WO 2023 / 056039, compounds disclosed in WO 2022 / 247816, and compounds disclosed in CN 115677688 A.

[0070] Compound 1, or pharmaceutically acceptable salts thereof, may be prepared using commercially available reagents and intermediates in the synthetic methods and reaction schemes described herein,WSGR Ref. 47134-779.601 those described in United States Patent No. 11,091,495, or may be prepared using other reagents and conventional methods well known to those skilled in the art. The contents of United States Patent No. 11,091,495 are hereby incorporated by reference forthat purpose.

[0071] In some embodiments are provided methods of treating cancer in a subject, comprising administering to the subject a pharmaceutically acceptable salt of Compound 1. The desired salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p -toluene sulfonic acid or ethanesulfonic acid, or the like.

[0072] If Compound 1, or a pharmaceutically acceptable salt thereof, is a solid, it is understood by those skilled in the art that the compounds or salts thereof may exist in different crystal or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulas.

[0073] Also provided herein are uses of isotopically -labeled Compound 1, or a pharmaceutically acceptable thereof, wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as2H and3H, carbon, such asnC,13C and14C, chlorine, such as36C1, fluorine, such as18F, iodine, such as123I and125I, nitrogen, such as13N and15N, oxygen, such as150,17O and18O, phosphorus, such as32P, and sulfur, such as35S. Certain isotopically-labeled compounds of the invention, for example, those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. The radioactive isotopes tritium (3H) and carbon-14 (14C) are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium,2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such asnC,18F,15O and13N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically- labeled Compound 1, or a pharmaceutically acceptable salt thereof, can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed.

[0074] In one aspect, the compositions described herein comprising Compound 1, or a pharmaceutically acceptable thereof, are used for the treatment of cancer in subjects. Such compositions may be prepared in pharmaceutically acceptable dosage forms for administration to subjects. Pharmaceutically acceptable dosage forms include, for example, liquids, suspensions, powders forWSGR Ref. 47134-779.601 reconstitution, tablets, pills, sachets, or capsules of hard or soft gelatin (See, e.g., Remington: The Science and Practice of Pharmacy (Gennaro, 21stEd. Mack Pub. Co., Easton, PA (2005)). Compound 1, or a pharmaceutically acceptable thereof, may be formulated into pharmaceutical compositions as described below in any pharmaceutical form recognizable to the skilled artisan as being suitable. Pharmaceutical compositions disclosed herein comprise a therapeutically effective amount of Compound 1, or a pharmaceutically acceptable thereof, and an inert, pharmaceutically acceptable carrier or diluent.

[0075] The pharmaceutical carriers employed may be either solid or liquid. Exemplary solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, and the like. Exemplary liquid carriers are syrup, peanut oil, olive oil, water, and the like. Similarly, the compositions may include time-delay or time-release material known in the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate or the like. Further additives or excipients may be added to achieve the desired formulation properties. For example, a bioavailability enhancer, such as Labrasol, Gelucire or the like, or formulator, such as CMC (carboxy-methylcellulose), PG (propyleneglycol), or PEG (polyethyleneglycol), may be added. Gelucire, a semi-solid vehicle that protects active ingredients from light, moisture, and oxidation, may be added, e.g., when preparing a capsule formulation.

[0076] If a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form, or formed into a troche or lozenge. The amount of solid carrier may vary, but generally will be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampoule or vial or non-aqueous liquid suspension. If a semi-solid carrier is used, the preparation may be in the form of hard and soft gelatin capsule formulations. The inventive compositions are prepared in unit-dosage form appropriate for the mode of administration, e.g., parenteral or oral administration.

[0077] To obtain a stable water-soluble dose form, Compound 1, or a pharmaceutically acceptable thereof, may be dissolved in an aqueous solution of an organic or inorganic acid, such as a 0.3 M solution of succinic acid or citric acid. If a soluble salt form is not available, the compound, or a pharmaceutically acceptable salt thereof, may be dissolved in a suitable co-solvent or combinations of co-solvents. Examples of suitable co-solvents include alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerin and the like in concentrations ranging from 0 to 60% of the total volume. In an exemplary embodiment, Compound 1, or a pharmaceutically acceptable salt thereof, is dissolved in DMSO and diluted with water. The composition may also be in the form of a solution of a salt form of the active ingredient in an appropriate aqueous vehicle such as water or isotonic saline or dextrose solution.

[0078] Proper formulation is dependent upon the route of administration selected. For injection, Compound 1, or a pharmaceutically acceptable thereof, may be formulated into aqueous solutions, preferably in physiologically compatible buffers such as Hanks solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.WSGR Ref. 47134-779.601

[0079] For oral administration, Compound 1, or a pharmaceutically acceptable thereof, can be formulated by combining the active compounds with pharmaceutically acceptable carriers known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by a subject to be treated. Pharmaceutical preparations for oral use can be obtained using a solid excipient in admixture with the active ingredient (agent), optionally grinding the resulting mixture, and processing the mixture of granules after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include: fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; and cellulose preparations, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum, methyl cellulose, hydroxypropylmethyl -cellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as crosslinked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0080] Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, polyvinyl pyrrolidone, Carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active agents.

[0081] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with fillers such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active agents may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0082] For administration intranasally or by inhalation, Compound 1, or a pharmaceutically acceptable thereof, may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of gelatin for use in an inhaler or insufflator and the like may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0083] Compound 1, or a pharmaceutically acceptable thereof, may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit-dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily orWSGR Ref. 47134-779.601 aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0084] Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active agents may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[0085] Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0086] Compound 1, or a pharmaceutically acceptable thereof, may also be formulated as a depot preparation. Such long-acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, Compound 1, or a pharmaceutically acceptable thereof, may be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion-exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt. A pharmaceutical carrier for hydrophobic compounds is a co-solvent system comprising benzyl alcohol, a non-polar surfactant, a water-miscible organic polymer, and an aqueous phase. The co-solvent system may be a VPD co-solvent system. VPD is a solution of 3% w / v benzyl alcohol, 8% w / v of the non-polar surfactant polysorbate 80, and 65% w / v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD: 5W) contains VPD diluted 1: 1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. The proportions of a co-solvent system may be suitably varied without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity non-polar surfactants may be used instead of polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may be substituted for dextrose.

[0087] Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide (DMSO) also may be employed, although usually at the cost of greater toxicity due to the toxic nature of DMSO. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemicalWSGR Ref. 47134-779.601 nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.

[0088] The pharmaceutical compositions also may comprise suitable solid- or gel-phase carriers or excipients. These carriers and excipients may provide marked improvement in the bioavailability of poorly soluble drugs. Examples of such carriers or excipients include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[0089] Further, the pharmaceutical composition may be incorporated into a skin patch for delivery of the drug directly onto the skin.

[0090] Additionally, the pharmaceutically acceptable formulations Compound 1, or a pharmaceutically acceptable thereof, that may be used to practice the methods disclosed herein may contain Compound 1, or a pharmaceutically acceptable thereof, in an amount of from about 0.5 w / w % to about 95 w / w %, or from about 1 w / w % to about 95 w / w %, or from about 1 w / w % to about 75 w / w %, or from about 5 w / w % to about 75 w / w %, or from about 10 w / w % to about 75 w / w %, or from about 10 w / w % to about 50 w / w %.

[0091] It will be appreciated that the actual dosages of Compound 1 , or a pharmaceutically acceptable thereof, to be administered to a subject in need thereof, will vary according to the particular composition of the formulation, the mode of administration, and the particular site, host, and disease being treated. Those skilled in the art using conventional dosage-determination tests in view of the experimental data for a given compound may ascertain optimal dosages for a given set of conditions. For oral administration, an exemplary daily dose of Compound 1, or a pharmaceutically acceptable thereof, generally employed will be from about 0.001 to about 1000 mg / kg of body weight, with courses of treatment repeated at appropriate intervals. In some embodiments are provided the methods disclosed herein, Compound 1, or a pharmaceutically acceptable thereof, is administered to the subject in an amount between about 0.01 mg / kg per day to about 300 mg / kg per day. In other embodiments are provided the methods disclosed herein, wherein Compound 1, or a pharmaceutically acceptable thereof, is administered to the subject in an amount between about 0.1 mg / kg per day to about 100 mg / kg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable thereof, is administered in an amount between about 10 mg to 500 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable thereof, is administered in an amount between about 100 mg to about 400 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable thereof, is administered in an amount between about 150 mg to about 350 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable thereof, is administered in an amount between about 150 mg to about 300 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered in an amount between about 160 mg to about 300 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered in an amount of about 160 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered in an amount of about 200 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof,WSGR Ref. 47134-779.601 is administered in an amount of about 240 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered in an amount of about 280 mg per day. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered in an amount of about 320 mg per day.

[0092] In some embodiments are provided the methods disclosed herein, wherein Compound 1, or a pharmaceutically acceptable salt thereof, is administered to the subject in an amount between about 100 mg and about 1000 mg once per day, between about 100 mg and about 900 mg once per day, between about 100 mg and about 850 mg once per day, between about 100 mg and about 800 mg once per day, between about 100 mg and about 750 mg once per day, between about 100 mg and about 700 mg once per day, between about 100 mg and about 650 mg once per day, between about 100 mg and about 600 mg once per day, between about 100 mg and about 550 mg once per day, or between about 100 mg and about 500 mg once per day.

[0093] In some embodiments are provided the methods disclosed herein, wherein Compound 1, or a pharmaceutically acceptable salt thereof, is administered to the subject in an amount of about 100 mg once per day, about 150 mg once per day, about 200 mg once per day, about 300 mg once per day, about 225 mg once per day, about 275 mg once per day, about 300 mg once per day, about 325 mg once per day, about 350 mg once per day, about 375 mg once per day, about 400 mg once per day, about 425 mg once per day, about 450 mg once per day, about 475 mg once per day, about 500 mg once per day, about 525 mg once per day, about 550 mg once per day, about 575 mg once per day, about 600 mg once per day, about 625 mg once per day, about 650 mg once per day, about 675 mg once per day, about 700 mg once per day, about 725 mg once per day, about 750 mg once per day, about 775 mg once per day, about 800 mg once per day, about 825 mg once per day, about 850 mg once per day, about 875 mg once per day, about 900 mg once per day, about 925 mg once per day, about 950 mg once per day, about 975 mg once per day, or about 1000 mg once per day.

[0094] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount between about 100 mg and about 1000 mg once per day, between about 100 mg and about 900 mg once per day, between about 100 mg and about 850 mg once per day, between about 100 mg and about 800 mg once per day, between about 100 mg and about 750 mg once per day, between about 100 mg and about 700 mg once per day, between about 100 mg and about 650 mg once per day, between about 100 mg and about 600 mg once per day, between about 100 mg and about 550 mg once per day, or between about 100 mg and about 500 mg once per day.

[0095] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 100 mg once per day, about 150 mg once per day, about 200 mg once per day, about 300 mg once per day, about 225 mg once per day, about 275 mg once per day, about 300 mg once per day, about 325 mg once per day, about 350 mg once per day, about 375 mg once per day, about 400 mg once per day, about 425 mg once per day, about 450 mg once per day, about 475 mg once per day, about 500 mg once per day, about 525 mg once per day, about 550 mg once perWSGR Ref. 47134-779.601 day, about 575 mg once per day, about 600 mg once per day, about 625 mg once per day, about 650 mg once per day, about 675 mg once per day, about 700 mg once per day, about 725 mg once per day, about 750 mg once per day, about 775 mg once per day, about 800 mg once per day, about 825 mg once per day, about 850 mg once per day, about 875 mg once per day, about 900 mg once per day, about 925 mg once per day, about 950 mg once per day, about 975 mg once per day, or about 1000 mg once per day.

[0096] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 100 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 200 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 400 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 500 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 600 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 700 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 800 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 900 mg once per day

[0097] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 24 hours following administration.

[0098] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma ofWSGR Ref. 47134-779.601 the subject of equal to or greater than 200 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 24 hours following administration.

[0099] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 24 hours following administration.

[0100] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal toWSGR Ref. 47134-779.601 or greater than 275 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 24 hours following administration

[0101] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 24 hours following administration.

[0102] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400WSGR Ref. 47134-779.601 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 24 hours following administration.

[0103] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 24 hours following administration.

[0104] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 16 hoursWSGR Ref. 47134-779.601 following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 24 hours following administration.

[0105] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 24 hours following administration.

[0106] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 20 hours following administration.WSGR Ref. 47134-779.601In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 24 hours following administration.

[0107] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 24 hours following administration.

[0108] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 24 hours following administration.WSGR Ref. 47134-779.601

[0109] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 24 hours following administration.

[0110] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 24 hours following administration.

[0111] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 4 hours following administration. In someWSGR Ref. 47134-779.601 embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 24 hours following administration.

[0112] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 24 hours following administration.

[0113] Furthermore, the pharmaceutically acceptable formulations of Compound 1, or a pharmaceutically acceptable salt thereof, that may be used to practice the methods disclosed herein may contain Compound 1, or a pharmaceutically acceptable salt thereof, in an amount of about 10 mg to about 2000 mg, or from about 10 mg to about 1500 mg, or from about 10 mg to about 1000 mg, or from about 10 mg to about 750 mg, or from about 10 mg to about 500 mg, or from about 25 mg to about 500 mg, or from about 50 mg to about 500 mg, or from about 100 mg to about 500 mg.WSGR Ref. 47134-779.601

[0114] In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof once a day. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof twice a day. Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof three times a day.

[0115] In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof in 28-day cycles. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof in multiple 28-day cycles. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof for at least one 28-day cycle. In some embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a subject in need thereof on each day of each 28-day cycle.

[0116] In some instances, the methods described herein comprise administering the compositions and formulations comprising Compound 1, or a pharmaceutically acceptable salt thereof, in combination with one or more additional therapeutic agents, to the subject or subject in need thereof in multiple cycles repeated on a regular schedule with periods of rest in between each cycle. For example, in some instances, treatment given for one week followed by three weeks of rest is one treatment cycle. The length of a treatment cycle depends on the treatment being given. In some embodiments, the length of a treatment cycle ranges from two to six weeks. In some embodiments, the length of a treatment cycle ranges from three to six weeks. In some embodiments, the length of a treatment cycle ranges from three to four weeks. In some embodiments, the length of a treatment cycle is three weeks (or 21 days). In some embodiments, the length of a treatment cycle is four weeks (28 days). In some embodiments, the length of a treatment cycle is 56 days. In some embodiments, a treatment cycle lasts one, two, three, or four weeks. In some embodiments, a treatment cycle lasts three weeks. In some embodiments, a treatment cycle lasts four weeks. The number of treatment doses scheduled within each cycle also varies depending on the drugs being given.

[0117] Dosages of compositions described herein can be determined by any suitable method. Maximum tolerated doses (MTD) and maximum response doses (MRD) for Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor, and, optionally, the additional therapeutic agents when administered to the subject, can be determined via established animal and human experimental protocols as well as in the examples described herein. For example, toxicity and therapeutic efficacy of Compound 1, or a pharmaceutically acceptable salt thereof, and the additional therapeutic agent can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between LD50 and ED50. The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in a human. TheWSGR Ref. 47134-779.601 dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with minimal toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. Additional relative dosages, represented as a percent of maximal response or of maximum tolerated dose, are readily obtained via the protocols.

[0118] In some embodiments, the amount of Compound 1, or a pharmaceutically acceptable salt thereof, and / or pharmaceutical formulations comprising them that corresponds to such an amount varies depending upon factors such as the particular salt or form, disease condition and its severity, the identity (e.g., age, weight, sex) of the subject or host in need of treatment, but can nevertheless be determined according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the liquid formulation type, the condition being treated, and the subject or host being treated.Methods of detecting biomarkers

[0119] The AKT inhibitor administered to the subject having cancer in combination Compound 1, or a pharmaceutically acceptable salt thereof, may be used in an amount according to the information known to those of ordinary skill in the art regarding those agents. For example, capivasertib may be administered to a subject according to the methods disclosed herein in amounts, and under conditions, according to those approved by one more regulatory authorities (e.g., the United States Food and Drug Administration) for the treatment of various types of cancer using those agents. In an embodiment, capivasertib is administered to a subject according to the methods described herein in an amount that is 400 mg twice daily, with or without food, for 4 days followed by 3 days off. In the event a subject experiences an adverse reaction in conjunction with the subject being administered capivasertib, the dose of capivasertib administered to such subject may be reduced to 320 mg twice daily for 4 days followed by 3 days off. In the event a second dose reduction of capivasertib is necessary, the dose of capivasertib administered to such subject may be reduced to 200 mg twice daily for 4 days followed by 3 days off. It is specifically contemplated herein that a subject administered Compound 1, or a pharmaceutically acceptable salt thereof, and capivasertib may require a reduction in the amount of capivasertib being administered to such subject while maintaining the same dose of Compound 1, or a pharmaceutically acceptable salt thereof. It is also specifically contemplated herein that a subject administered Compound 1, or a pharmaceutically acceptable salt thereof, and capivasertib may require a reduction in the amount of Compound 1, or a pharmaceutically acceptable salt thereof, while maintaining the same dose of capivasertib being administered to such subject. It is further contemplated herein that a subject administered Compound 1, or a pharmaceutically acceptable salt thereof, and capivasertib may require a reduction in the amount of Compound 1, or a pharmaceutically acceptable salt thereof, and a reduction in the amount of capivasertib.

[0120] In some embodiments are provided the methods disclosed herein wherein the measurement of one or more biomarkers is used to determine whether a subject would benefit from the administration Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor, or whether the subjectWSGR Ref. 47134-779.601 is responding to the administration of Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. Biomarkers that may be used according to the methods disclosed herein are those known to those having ordinary skill in the art, for example the presence of one or more PI3KA / AKT1 / PTEN alterations. In an embodiment, the biomarkers that may be used according to the methods disclosed herein include the presence of one or more PI3KA / AKT1 / PTEN alterations as detected by an FDA -approved test. In some embodiments, the biomarker comprises the presence of one or more PI3KA alterations in one or more cells comprising the cancer in the subject. In other embodiments, the biomarker comprises the presence of one or more AKT1 alterations in one or more cells comprising the cancer in the subject, the biomarker comprises the presence of one or more PTEN alterations in one or more cells comprising the cancer in the subject. In some embodiments, one or more cells comprising the cancer in the subject are determined to be PTEN-null.

[0121] Disclosed herein are methods comprising: (a) providing a biologic sample obtained from a subject having cancer; (b) assaying to detect in the biologic sample obtained from the subject a presence or absence of a biomarker; (c) detecting the presence or absence of the biomarker in the biologic sample using the methods described herein; and (d) administering to the subject a therapeutically effective amount of amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor if the biomarker is present in the biological sample. In some embodiments, the presence of one or more biomarkers described herein indicate the cancer in the subject is resistant to one or more prior therapies, such as the cancer being resistant to one or more chemotherapeutic agents or one or endocrine -based regimen.

[0122] The presence, absence, or level, of such biomarkers may be measured, collectively or individually, in a biological sample obtained from a subject, such as a sample of a solid tumor, such as breast cancer, or from a sample of a relevant biological fluid, such as a blood sample. In some instances, the one or more biomarkers are detected in plasma or serum that is derived from a blood sample obtained from the subject. In some instances, the methods of detection disclosed herein are useful for predicting a therapeutic response to a therapy described herein (e.g., the administration to a subject of amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor), monitor the treatment using the therapy of, and treating with the therapy, a proliferative disease or condition described herein in a subject.

[0123] In some embodiments, the expression of a biomarker in a biological sample from a subject is measured by use of immunohistochemistry (IHC) assays. Such immunohistochemistry (IHC) assays are commercially available, or may be developed and utilized according to methods known to those having ordinary skill in the art.

[0124] Immunohistochemistry techniques utilize an antibody to probe and visualize cellular antigens in situ, generally by chromogenic or fluorescent methods. In such techniques, antibodies or antisera, polyclonal antisera, or monoclonal antibodies specific for each marker are used to detect expression. The antibodies can be detected by direct labeling of the antibodies themselves, for example, with radioactiveWSGR Ref. 47134-779.601 labels, fluorescent labels, hapten labels such as, biotin, or an enzyme such as horse radish peroxidase or alkaline phosphatase. Alternatively, unlabeled primary antibody is used in conjunction with a labeled secondary antibody, comprising antisera, polyclonal antisera, or a monoclonal antibody specific for the primary antibody. Immunohistochemistry protocols and kits are well known in the art and are commercially available.

[0125] Two general methods of IHC are generally available; direct and indirect assays. According to the first assay, binding of antibody to the target antigen is determined directly. This direct assay uses a labeled reagent, such as a fluorescent tag or an enzyme-labeled primary antibody, which can be visualized without further antibody interaction. In a typical indirect assay, unconjugated primary antibody binds to the antigen and then a labeled secondary antibody binds to the primary antibody. Where the secondary antibody is conjugated to an enzymatic label, a chromagenic or Anorogenic substrate is added to provide visualization of the antigen. Signal amplification occurs because several secondary antibodies may react with different epitopes on the primary antibody. The primary and / or secondary antibody used for immunohistochemistry typically will be labeled with a detectable moiety. Numerous labels are available which can be generally grouped into the following categories. First, are radioisotopes, such as35S,14C,1251,3H, and131I. The antibody can be labeled with the radioisotope using the techniques described in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-Interscience, New York, N.Y., Pubs. (1991) for example and radioactivity can be measured using scintillation counting. Next, are colloidal gold particles. Third are fluorescent labels including, but are not limited to, rare earth chelates (europium chelates), Texas Red, rhodamine, fluorescein, dansyl, Lissamine, umbelliferone, phycocrytherin, phycocyanin, or commercially available fluorophores such SPECTRUM ORANGE® and SPECTRUM GREEN® and / or derivatives of any one or more of the above. The fluorescent labels can be conjugated to the antibody using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a fluorimeter. Fourth are various enzyme-substrate labels are available and U.S. Pat. No. 4,275,149 provides a review of some of these. The enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above. The chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor. Examples of enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3 -dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, [3-galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose -6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like. Techniques for conjugating enzymes to antibodies are described inWSGR Ref. 47134-779.601O'Sullivan et al. Methods for the Preparation of Enzyme-Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (ed J. Langone & H. Van Vunakis), Academic press, New York, 73: 147-166 (1981). Examples of enzyme-substrate combinations include, for example (i) Horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate, wherein the hydrogen peroxidase oxidizes a dye precursor [e.g., orthophenylene diamine (OPD) or 3,3 ’,5,5 ’-tetramethyl benzidine hydrochloride (TMB)]. 3,3 -Diaminobenzidine (DAB) may also be used to visualize the HRP -labeled antibody; (ii) alkaline phosphatase (AP) with para-Nitrophenyl phosphate as chromogenic substrate; and (iii) [3-D- galactosidase ( -D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl- -D-galactosidase) or Anorogenic substrate (e.g., 4-methylumbelliferyl-P-D-galactosidase). Numerous other enzyme-substrate combinations are available to those skilled in the art. These methods are generally described in U.S. Pat. Nos. 4,275,149 and 4,318,980. Sometimes, the label is indirectly conjugated with the antibody. The skilled artisan will be aware of various techniques for achieving this. For example, the antibody can be conjugated with biotin and any of the four broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the antibody in this indirect manner. Alternatively, to achieve indirect conjugation of the label with the antibody, the antibody is conjugated with a small hapten and one of the different types of labels mentioned above is conjugated with an anti-hapten antibody. Thus, indirect conjugation of the label with the antibody can be achieved.

[0126] Biological samples obtained from subjects comprising tissue samples may be prepared according to protocols commonly used in the art. Typically, sections of paraffin-embedded cells or tissues are obtained by (1) preserving tissue in fixative, (2) dehydrating the fixed tissue, (3) infiltrating the tissue with fixative, (4) orienting the tissue such that the cut surface accurately represents the tissue, (5) embedding the tissue in paraffin (making a paraffin block), (6) cutting tissue paraffin block with a microtome in sections of 4-5 picometers, and (7) mounting sections onto slides. The slides may then be read by a pathologist or the like assessing for the presence or absence of a biomarker, or of abnormal or normal cells or a specific cell type and provides the loci of the cell types of interest. Thus, for example, a pathologist or the like would review the slides and identify normal cells and abnormal cells (such as abnormal or tumor cells). Any means of defining the loci of the cells of interest may be used (e.g., coordinates on an X-Y axis).

[0127] Aside from the sample preparation procedures discussed above, further treatment of the tissue section prior to, during or following IHC may be desired. For example, epitope retrieval methods, such as heating the tissue sample in citrate buffer may be carried out [see, e.g., Leong et al. Appl.Immunohistochem. 4(3) :201 (1996)]. Following an optional blocking step, the tissue section is exposed to primary antibody for a sufficient period of time and under suitable conditions such that the primary antibody binds to the target protein antigen in the tissue sample. Appropriate conditions for achieving this can be determined by routine experimentation.WSGR Ref. 47134-779.601

[0128] The extent of binding of antibody to the sample is determined by using any one of the detectable labels discussed above. For example, the label is an enzymatic label (e.g., HRPO) which catalyzes a chemical alteration of the chromogenic substrate such as 3,3 ’-diaminobenzidine chromogen. Preferably the enzymatic label is conjugated to antibody which binds specifically to the primary antibody (e.g., the primary antibody is rabbit polyclonal antibody and secondary antibody is goat anti-rabbit antibody). Specimens thus prepared may be mounted and coverslipped. Slide evaluation is then determined, e.g., using a microscope.

[0129] IHC may be combined with morphological staining, either prior to or thereafter. After deparaffmization, the sections mounted on slides may be stained with a morphological stain for evaluation. The morphological stain to be used provides for accurate morphological evaluation of a tissue section. The section may be stained with one or more dyes each of which distinctly stains different cellular components. In one embodiment, hematoxylin is use for staining cellular nucleic of the slides. Hematoxylin is widely available. An example of a suitable hematoxylin is Hematoxylin II (Ventana). When lighter blue nuclei are desired, a bluing reagent may be used following hematoxylin staining. One of skill in the art will appreciate that staining may be optimized for a given tissue by increasing or decreasing the length of time the slides remain in the dye.

[0130] Automated systems for slide preparation and IHC processing are available commercially. The Ventana® BenchMark XT system is an example of such an automated system.

[0131] After staining, the tissue section may be analyzed by standard techniques of microscopy. Generally, a pathologist or the like assesses the tissue for the presence of abnormal or normal cells or a specific cell type and provides the loci of the cell types of interest. Thus, for example, a pathologist or the like would review the slides and identify normal cells and abnormal cells (such as abnormal or tumor cells). Any means of defining the loci of the cells of interest may be used (e.g., coordinates on an X-Y axis).

[0132] In some embodiments, the presence, or an absence, and / or a level of expression of the biomarker is detected in the sample obtained from a subject by analyzing the genetic material in the sample. In some embodiments, the genetic material is obtained from blood, serum, plasma, sweat, hair, tears, urine, and other techniques known by one of skill in the art. In some embodiments the sample comprises circulating tumor RNA (ctRNA). In some embodiments the sample comprises peripheral blood mononuclear cells (PBMCs). In some embodiments the sample comprises circulating tumor cells (CTCs). In some cases, the genetic material is obtained from a tumor biopsy or liquid biopsy. In some embodiments, a tumor biopsy comprises a formalin-fixed paraffin embedded biopsy, a fresh frozen biopsy, a fresh biopsy, or a frozen biopsy. In some embodiments, a liquid biopsy comprises PBMCs, circulating tumor RNA, plasma cell- free RNA, or circulating tumor cells (CTCs). Tumor and liquid biopsies can undergo additional analytic processing for sample dissociation, cell sorting, and enrichment of cell populations of interest.

[0133] In some embodiments, methods of detecting a presence, absence, or level of a biomarker in a biologic sample obtained from the subject involve detecting a nucleic acid sequence. In some cases, theWSGR Ref. 47134-779.601 nucleic acid sequence comprises deoxyribonucleic acid (DNA), such as in the case of detecting complementary DNA (cDNA) of an mRNA transcript. In some instances, the nucleic acid sequence comprises a denatured DNA molecule or fragment thereof. In some instances, the nucleic acid sequence comprises DNA selected from: genomic DNA, viral DNA, mitochondrial DNA, plasmid DNA, amplified DNA, circular DNA, circulating DNA, cell-free DNA, or exosomal DNA. In some instances, the DNA is single -stranded DNA (ssDNA), double-stranded DNA, denaturing double-stranded DNA, synthetic DNA, and combinations thereof. The circular DNA may be cleaved or fragmented. In some instances, the nucleic acid sequence comprises ribonucleic acid (RNA). In some instances, the nucleic acid sequence comprises fragmented RNA. In some instances, the nucleic acid sequence comprises partially degraded RNA. In some instances, the nucleic acid sequence comprises a microRNA or portion thereof. In some instances, the nucleic acid sequence comprises an RNA molecule or a fragmented RNA molecule (RNA fragments) selected from: a microRNA (miRNA), a pre-miRNA, a pri-miRNA, a mRNA, a pre-mRNA, a viral RNA, a viroid RNA, a virusoid RNA, circular RNA (circRNA), a ribosomal RNA (rRNA), a transfer RNA (tRNA), a pre-tRNA, a long non-coding RNA (IncRNA), a small nuclear RNA (snRNA), a circulating RNA, a cell-free RNA, an exosomal RNA, a vector-expressed RNA, an RNA transcript, a synthetic RNA, and combinations thereof.

[0134] Disclosed herein, in some embodiments, a biomarker is detected by subjecting a sample obtained from the subject to a nucleic acid-based detection assay. In some instances, the nucleic acid-based detection assay comprises quantitative polymerase chain reaction (qPCR), reverse transcription PCT (RT- qPCR), gel electrophoresis (including for e.g., Northern or Southern blot), immunohistochemistry (IHC), immunofluorescence (IF), in situ hybridization (ISH) such as fluorescent in situ hybridization (FISH), cytochemistry, microarray, or sequencing. In some embodiments, the sequencing technique comprises next generation sequencing. In some embodiments, the methods involve a hybridization assay such as Anorogenic qPCR (e.g., TaqMan™, SYBR green, SYBR green I, SYBR green II, SYBR gold, ethidium bromide, methylene blue, Pyronin Y, DAPI, acridine orange, Blue View or phycoerythrin), which involves a nucleic acid amplification reaction with a specific primer pair, and hybridization of the amplified nucleic acid probes comprising a detectable moiety or molecule that is specific to a target nucleic acid sequence. In some instances, a number of amplification cycles for detecting a target nucleic acid in a qPCR assay is about 5 to about 30 cycles. In some instances, the number of amplification cycles for detecting a target nucleic acid is at least about 5 cycles. In some instances, the number of amplification cycles for detecting a target nucleic acid is at most about 30 cycles. In some instances, the number of amplification cycles for detecting a target nucleic acid is about 5 to about 10, about 5 to about 15, about 5 to about 20, about 5 to about 25, about 5 to about 30, about 10 to about 15, about 10 to about 20, about 10 to about 25, about 10 to about 30, about 15 to about 20, about 15 to about 25, about 15 to about 30, about 20 to about 25, about 20 to about 30, or about 25 to about 30 cycles. For TaqMan™ methods, the probe may be a hydrolysable probe comprising a fluorophore and quencher that is hydrolyzed by DNA polymerase when hybridized to a target nucleic acid. In some cases, the presence of a target nucleic acid isWSGR Ref. 47134-779.601 determined when the number of amplification cycles to reach a threshold value is less than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, or 20 cycles. In some instances, hybridization may occur at standard hybridization temperatures, e.g., between about 35 °C and about 65 °C in a standard PCR buffer.

[0135] An additional exemplary nucleic acid-based detection assay comprises the use of nucleic acid probes conjugated or otherwise immobilized on a bead, multi-well plate, or other substrate, wherein the nucleic acid probes are configured to hybridize with a target nucleic acid sequence. In some instances, the nucleic acid probe is specific to one or more of a polynucleotide sequence that encodes a relevant biomarker as disclosed herein. In some instances, the nucleic acid probe specific to a biomarker comprises a nucleic acid probe sequence sufficiently complementary to the polynucleotide sequence that encodes the relevant biomarker protein. In some instances, the probe comprises a transcribed polynucleotide sequence (e.g., RNA, cDNA). In some embodiments, the nucleic acid probe can be, for example, a full-length cDNA, or a portion thereof, such as an oligonucleotide of at least about 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides in length and sufficient to specifically hybridize under standard hybridization conditions to the target nucleic acid sequence. In some embodiments, the target nucleic acid sequence is immobilized on a solid surface and contacted with a probe, for example by running the isolated target nucleic acid sequence on an agarose gel and transferring the target nucleic acid sequence from the gel to a membrane, such as nitrocellulose. In some embodiments, the probe(s) are immobilized on a solid surface, for example, in an Affymetrix gene chip array, and the probe(s) are contacted with the target nucleic acid sequence.

[0136] In some embodiments, the term “probe” with regards to nucleic acids, refers to any nucleic acid molecule that is capable of selectively binding to a specifically intended target nucleic acid sequence. In some instances, probes are specifically designed to be labeled, for example, with a radioactive label, a fluorescent label, an enzyme, a chemiluminescent tag, a colorimetric tag, or other labels or tags that are known in the art. In some instances, the fluorescent label comprises a fluorophore. In some instances, the fluorophore is an aromatic or heteroaromatic compound. In some instances, the fluorophore is a pyrene, anthracene, naphthalene, acridine, stilbene, benzoxazole, indole, benzindole, oxazole, thiazole, benzothiazole, canine, carbocyanine, salicylate, anthranilate, xanthenes dye, coumarin. Exemplary xanthene dyes include, e.g., fluorescein and rhodamine dyes. Fluorescein and rhodamine dyes include, but are not limited to 6-carboxyfluorescein (FAM), 2'7'-dimethoxy-4'5'-dichloro-6-carboxyfluorescein (JOE), tetrachlorofluorescein (TET), 6-carboxyrhodamine (R6G), N,N,N; N'-tetramethyl-6-carboxyrhodamine (TAMRA), 6-carboxy-X-rhodamine (ROX). Suitable fluorescent probes also include the naphthylamine dyes that have an amino group in the alpha or beta position. For example, naphthylamino compounds include I-dimethylaminonaphthyl-5-sulfonate, l-anilino-8-naphthalene sulfonate, and 2-p-toluidinyl-6- naphthalene sulfonate, 5-(2'-aminoethyl)aminonaphthalene-I-sulfonic acid (EDANS). Exemplary coumarins include, e.g., 3-phenyl-7-isocyanatocoumarin; acridines, such as 9-isothiocyanatoacridine and acridine orange; N-(p-(2-benzoxazolyl)phenyl) maleimide; cyanines, such as, e.g., indodicarbocyanine 3 (Cy3), indodicarbocyanine 5 (Cy5), indodicarbocyanine 5.5 (Cy5.5), 3-(-carboxy-pentyl)-3'-ethyl-5,5'-WSGR Ref. 47134-779.601 dimethyloxacarbocyanine (CyA); 1H, 5H, 11H, 15H-Xantheno[2,3, 4-ij: 5,6, 7-i'j']diquinolizin-18-ium, 9- [2 (or 4)-[[[6-[2,5-dioxo-l-pyrrolidinyl)oxy]-6-oxohexyl]amino]sulfonyl]-4 (or 2)-sulfophenyl]-2,3, 6,7, 12,13, 16,17-octahydro-inner salt (TR or Texas Red); or BODIPYTM dyes. In some cases, the probe comprises FAM as the dye label.

[0137] In some embodiments, detecting the one or more biomarkers comprises sequencing genetic material obtained from a sample from the subject. Sequencing can be performed with any appropriate sequencing technology, including but not limited to single -molecule real-time (SMRT) sequencing, Polony sequencing, sequencing by ligation, reversible terminator sequencing, proton detection sequencing, ion semiconductor sequencing, nanopore sequencing, electronic sequencing, pyrosequencing, Maxam-Gilbert sequencing, chain termination (e.g., Sanger) sequencing, +S sequencing, or sequencing by synthesis. Sequencing methods also include next-generation sequencing, e.g., modem sequencing technologies such as Illumina sequencing (e.g., Solexa), Roche 454 sequencing, Ion torrent sequencing, and SOLiD sequencing. In some cases, next-generation sequencing involves high-throughput sequencing methods. Additional sequencing methods available to one of skill in the art may also be employed.

[0138] In some instances, a number of nucleotides that are sequenced are at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 300, 400, 500, 2000, 4000, 6000, 8000, 10000, 20000, 50000, 100000, or more than 100000 nucleotides. In some instances, the number of nucleotides sequenced is in a range of about 1 to about 100000 nucleotides, about 1 to about 10000 nucleotides, about 1 to about 1000 nucleotides, about 1 to about 500 nucleotides, about 1 to about 300 nucleotides, about 1 to about 200 nucleotides, about 1 to about 100 nucleotides, about 5 to about 100000 nucleotides, about 5 to about 10000 nucleotides, about 5 to about 1000 nucleotides, about 5 to about 500 nucleotides, about 5 to about 300 nucleotides, about 5 to about 200 nucleotides, about 5 to about 100 nucleotides, about 10 to about 100000 nucleotides, about 10 to about 10000 nucleotides, about 10 to about 1000 nucleotides, about 10 to about 500 nucleotides, about 10 to about 300 nucleotides, about 10 to about 200 nucleotides, about 10 to about 100 nucleotides, about 20 to about 100000 nucleotides, about 20 to about 10000 nucleotides, about 20 to about 1000 nucleotides, about 20 to about 500 nucleotides, about 20 to about 300 nucleotides, about 20 to about 200 nucleotides, about 20 to about 100 nucleotides, about 30 to about 100000 nucleotides, about 30 to about 10000 nucleotides, about 30 to about 1000 nucleotides, about 30 to about 500 nucleotides, about 30 to about 300 nucleotides, about 30 to about 200 nucleotides, about 30 to about 100 nucleotides, about 50 to about 100000 nucleotides, about 50 to about 10000 nucleotides, about 50 to about 1000 nucleotides, about 50 to about 500 nucleotides, about 50 to about 300 nucleotides, about 50 to about 200 nucleotides, or about 50 to about 100 nucleotides.

[0139] In some cases, a hybridization assay, such as those described herein, is used to detect the mRNA encoding the biomarker in the sample. Exemplary probe sequences that are hybridizable to a target nucleic acid sequence comprise at least 10, but no more than 100 contiguous nucleotides comprising the relevant sequence. In some cases, RNA sequencing (RNAseq) is used to detect the mRNA encoding the relevant biomarker protein.WSGR Ref. 47134-779.601

[0140] Detection of the mRNA, in some cases, involves amplification of the subject’s nucleic acid by the polymerase chain reaction (PCR). In some embodiments, the PCR assay involves use of a pair of primers capable of amplifying at least about 10 contiguous nucleobases within a nucleic acid sequence, thereby amplifying the one or more gene products in the biomarker. In Anorogenic quantitative PCR, quantitation is based on amount of Huorescence signals (TaqMan and SYBR green). In some embodiments, the nucleic acid probe is conjugated to a detectable molecule. The detectable molecule may be a Huorophore. The nucleic acid probe may also be conjugated to a quencher.

[0141] In some embodiments, the assay for detecting the presence or absence of mRNA encoding a relevant biomarker comprises reverse-transcribing the relevant mRNA molecule to produce a corresponding complementary DNA (cDNA) molecule. In some embodiments, the assay further comprises contacting the cDNA molecule with a nucleic acid probe comprising a nucleic acid sequence that is complementary to a nucleic acid sequence of the cDNA molecule. In some embodiments, the assay comprises detecting a double-stranded hybridization product between the nucleic acid probe and the cDNA molecule. In some embodiments, the hybridization product is further amplified using a pair of primers. In some embodiments, the primers comprises a first primer with a nucleic acid sequence comprising at least 10 but not more than 50 contiguous nucleic acids within a relevant nucleic acid sequence that binds to a top strand of the double -stranded hybridization product; and a second primer with a nucleic acid sequence comprising at least 10 but not more than 50 contiguous nucleic acids within a nucleic acid sequence that is reverse complement to the relevant nucleic acid sequence that binds to a bottom strand of the double -stranded hybridization product.

[0142] Disclosed herein, in some embodiments, are methods comprising preparing a complementary DNA (cDNA) library. In some embodiments, the cDNA library is sequenced using suitable sequence methodologies disclosed herein. In some embodiments, the cDNA library is labeled, a plurality of nucleic acid probes is generated, and fixed to an immobile surface (such as a microarray). In some embodiments, the plurality of nucleic acid probes is capable of hybridizing to at least about 10 contiguous nucleotides of the two or more genes in a sample obtained from the subject. In some embodiments, detecting the presence of or absence of a biomarker includes detecting a high or a low level of expression of one or more genes as compared to a reference level.

[0143] Disclosed herein, in some embodiments, genetic material is extracted from a biologic sample obtained from a subject, e.g., a sample of blood, serum, or tissue. In certain embodiments where nucleic acids are extracted, the nucleic acids are extracted using any technique that does not interfere with subsequent analysis. In certain embodiments, this technique uses alcohol precipitation using ethanol, methanol, or isopropyl alcohol. In certain embodiments, this technique uses phenol, chloroform, or any combination thereof. In certain embodiments, this technique uses cesium chloride. In certain embodiments, this technique uses sodium, potassium or ammonium acetate or any other salt commonly used to precipitate DNA. In certain embodiments, this technique utilizes a column or resin based nucleic acid purification scheme such as those commonly sold commercially, one non-limiting example would beWSGR Ref. 47134-779.601 the GenElute Bacterial Genomic DNA Kit available from Sigma Aldrich. In certain embodiments, after extraction the nucleic acid is stored in water, Tris buffer, or Tris-EDTA buffer before subsequent analysis. In an exemplary embodiment, the nucleic acid material is extracted in water. In some cases, extraction does not comprise nucleic acid purification. In certain embodiments, RNA may be extracted from cells using RNA extraction techniques including, for example, using acid phenol / guanidine isothiocyanate extraction (RNAzol B; Biogenesis), RNeasy RNA preparation kits (Qiagen) or PAXgene (PreAnalytix, Switzerland).

[0144] In some aspects, circulating tumor RNA (ctRNA) is used to assess the expression levels of RNA molecules, shed by the tumor into the blood stream. In some embodiments, detection of ctRNA is useful, for example, for detecting and diagnosing a tumor. Because tumor DNA and RNA has acquired multiple genetic mutations, leading to tumor development, ctRNA are not an exact match to the individual’s DNA and RNA, respectively. Finding DNA and RNA with genetic differences aids in tumor detection. Diagnosing the type of tumor using ctRNA can reduce the need for getting a sample of the tumor tissue (tumor biopsy), which can be challenging when a tumor is difficult to access, such as a tumor in the brain or lung.

[0145] In some embodiments, a decrease in the quantity of ctRNA suggests the solid tumor is shrinking and treatment of the subject with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor is effective. In some embodiments, a lack of ctRNA in the bloodstream indicates that the cancer has not returned after treatment of the subject with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor.

[0146] Described herein are methods of assessing genetic alterations by ctRNA profiling. In some embodiments, the genomic profiling is performed after each treatment cycle with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the gene alterations indicate that the cancer is becoming resistant to the treatment with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the lack of gene alterations indicate that the cancer is not becoming resistant to the treatment with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor.

[0147] Described herein are methods of assessing genetic alterations, including, but not limited to, mutations in certain genes and / or copy number alterations in certain genes, by circulating tumor DNA (ctDNA) and / or cell-free DNA (cfDNA) profiling. In some embodiments, the genomic profiling is performed after each treatment cycle with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the gene alterations indicate that the cancer is becoming resistant to the treatment with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the lack of gene alterations indicate that the cancer is not becoming resistant to the treatment with Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor.WSGR Ref. 47134-779.601

[0148] In some embodiments, the expression of a biomarker is measured by immunofluorescence (IF) assays. In some embodiments, the expression of a biomarker is measured by in situ hybridization (ISH) assays. In some embodiments, the expression of a biomarker transcript levels are measured using assays such as quantitative polymerase chain reaction (qPCR), microarray, and RNA sequencing, or assays commercially available from companies such as Fluidigm and Nanostring.

[0149] Disclosed herein are methods of treating a subject having cancer, comprising: (a) providing a biologic sample obtained from a subject having cancer; (b) assaying to detect in the biologic sample obtained from the subject a presence or absence of a biomarker; (c) detecting the presence or absence of the biomarker in the biologic sample using the methods described herein; and (d) administering to the subject a therapeutically effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor if the biomarker is present in the biological sample. In some embodiments, expression of a biomarker is based on the expression level of the biomarker deviating from a reference expression level. In some embodiments, the expression level is high, relative to the reference expression level. In some embodiments, the expression level is low, relative to the reference expression level. In some embodiments, the reference expression level is derived from an individual, or a group of individuals, that do not have cancer. In some embodiments, the reference expression level is derived from an individual, or a group of individuals, that have cancer that does not therapeutically respond to the administration of Compound 1, or a pharmaceutically acceptable salt thereof, and an AKT inhibitor. In some embodiments, the expression level deviates from the reference expression level by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.

[0150] In some embodiments, the determination of expression or the presence of a biomarker is defined based on the percentage of cells that stain weakly, moderately, or strongly for the relevant biomarker, with the threshold defining the minimal percentage of cells that are required to stain positive at the various intensity levels (>a% of tumor cells stain weakly, >b% of tumor cells stain moderately, >c% of tumor cells stain strongly, or a combination thereof). In some embodiments, the one or more of the cells comprising the cancer has been determined to express a biomarker when > about 10%, > about 15%, > about 20%, > about 25%, > about 30%, > about 35%, > about 40%, > about 45%, > about 50%, > about 55%, > about 60%, > about 65%, > about 70%, > about 75%, > about 80%, > about 85%, > about 90%, or > about 95% of the tumor cells stain weakly for the biomarker; when > about 10%, > about 15%, > about 20%, > about 25%, > about 30%, > about 35%, > about 40%, > about 45%, > about 50%, > about 55%, > about 60%, > about 65%, > about 70%, > about 75%, > about 80%, > about 85%, > about 90%, or > about 95% of the tumor cells stain moderately for the biomarker; when > about 10%, > about 15%, > about 20%, > about 25%, > about 30%, > about 35%, > about 40%, > about 45%, > about 50%, > about 55%, > about 60%, > about 65%, > about 70%, > about 75%, > about 80%, > about 85%, > about 90%, or > about 95% of the tumor cells stain strongly for the biomarker; or any combinations thereof.WSGR Ref. 47134-779.601Kits and articles of manufacture

[0151] Disclosed herein, in certain embodiments, are kits and articles of manufacture for use with one or more methods and compositions described herein. Such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. In one embodiment, the containers are formed from a variety of materials such as glass or plastic.

[0152] A kit typically includes labels listing contents and / or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.

[0153] In one embodiment, a label is on or associated with the container. In one embodiment, a label is on a container when letters, numbers or other characters forming the label are attached, molded, or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. In one embodiment, a label is used to indicate that the contents are to be used for a specific therapeutic application. The label also indicates directions for use of the contents, such as in the methods described herein.

[0154] In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack, for example, contains metal or plastic foil, such as a blister pack. In one embodiment, the pack or dispenser device is accompanied by instructions for administration. In one embodiment, the pack or dispenser is also accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for drugs, or the approved product insert. In one embodiment, compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0155] Disclosed herein is a kit comprising (a) Compound 1, or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor, for use in treating cancer in a subject in need thereof and a package insert comprising instructions for determining when the administration to the subject having cancer measuring the expression of a biomarker described herein in one or more of the cells comprising the cancer and (a) Compound 1, or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor if one or more of the cells comprising the cancer has been determined to express the biomarker.WSGR Ref. 47134-779.601Numbered Embodiments

[0156] Embodiment 1. A method of treating cancer in a subject, comprising administering to the subject(a) Compound(Compound 1), or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor.

[0157] Embodiment 2. A method of treating cancer in a subject, wherein the cancer in the subject has been determined to comprise one or more PIK3CA / AKTl / PTEN-alterations, comprising administering to the subject (a) Compound 1, or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor.

[0158] Embodiment 3. The method of embodiment 2, wherein the cancer in the subject has been determined to comprise one or more PIK3CA alterations.

[0159] Embodiment 4. The method of embodiment 2, wherein the cancer in the subject has been determined to comprise one or more AKT1 alterations.

[0160] Embodiment 5. The method of embodiment 2, wherein the cancer in the subject has been determined to comprise one or more PTEN alterations.

[0161] Embodiment 6. The method of embodiment 5, wherein the cancer in the subject has been determined to be PTEN-null.

[0162] Embodiment 7. The method of any one of embodiments 1 to 6, wherein the subject has received prior administration of one or more chemotherapeutic agents prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

[0163] Embodiment 8. The method of embodiment 7, wherein the subject has received up to one chemotherapeutic treatments prior the administration to the subject prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

[0164] Embodiment 9. The method of any one of embodiments 1 to 6, wherein the subject has not been administered a chemotherapeutic agent prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

[0165] Embodiment 10. The method of any one of embodiments 1 to 9, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject sequentially or simultaneously.

[0166] Embodiment 11. The method of embodiment 10, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject sequentially.

[0167] Embodiment 12. The method of embodiment 11, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject on the same day.WSGR Ref. 47134-779.601

[0168] Embodiment 13. The method of embodiment 11, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject within a 24-hour period.

[0169] Embodiment 14. The method of embodiment 10, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject simultaneously.

[0170] Embodiment 15. The method of any one of embodiments 1 to 14, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject once per day or twice per day.

[0171] Embodiment 16. The method of embodiment 15, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject once per day.

[0172] Embodiment 17. The method of embodiment 15, wherein Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor are administered to the subject twice per day.

[0173] Embodiment 18. The method of any one of embodiments 1 to 14, wherein Compound 1, or a pharmaceutically acceptable salt thereof, is administered to the subject once per day and the AKT inhibitor is administered to the subject twice per day.

[0174] Embodiment 19. The method of any one of embodiments 1 to 18, wherein the subject has been administered at least one endocrine -based therapy prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

[0175] Embodiment 20. The method of embodiment 19, wherein the cancer in the subject has progressed on the at least one endocrine-based therapy prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

[0176] Embodiment 21. The method of any one of embodiments 1 to 20, wherein the cancer in the subject is selected from diffused large B cell lymphoma (DLBCL), follicular lymphoma, other lymphomas, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, cancer, breast cancer, bile duct and gallbladder cancers, bladder carcinoma, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal carcinoma, ovarian cancer, pancreatic cancer, renal cell carcinoma, prostate cancer, rectal cancer, thyroid cancers, parathyroid tumors, uterine tumors, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), Kaposi sarcoma, synovial sarcoma, osteosarcoma and Ewing's sarcoma.

[0177] Embodiment 22. The method of embodiment 21, wherein the cancer in the subject is breast cancer.

[0178] Embodiment 23. The method of embodiment 22, wherein the breast cancer in the subject is selected from HR-positive breast carcinoma, HR-negative breast carcinoma, primary breast ductal carcinoma, mammary adenocarcinoma, HR-positive mammary ductal carcinoma, HR- negative mammary ductal carcinoma, HER2 -positive mammary ductal carcinoma, HER2 -positive breast cancer, luminal breast cancer, triple negative breast cancer (TNBC), ER-positive breast cancer, ER-negative breast cancer, PR-positive breast cancer, PR-negative breast cancer, HR-positive / HER-2 negative breast cancer, HR-WSGR Ref. 47134-779.601 negative / HER-2 negative breast cancer, HR-positive / HER-2 positive breast cancer, and HR- negative / HER-2 positive breast cancer, and unclassified breast cancer.

[0179] Embodiment 24. The method of embodiment 22, wherein the breast cancer in the subject is HR- positive / HER-2 negative breast cancer.

[0180] Embodiment 25. The method of embodiment 22, wherein the breast cancer in the subject is HR- positive / HER-2 negative, locally advanced or metastatic breast cancer.

[0181] Embodiment 26. Embodiment 20. The method of embodiment 22, wherein the breast cancer in the subject is HR-positive / HER-2 negative, locally advanced or metastatic breast cancer, with one or more PIK3 CA / AKT 1 / PTEN -alterations .

[0182] Embodiment 27. The method of embodiment 22, wherein the breast cancer in the subject is HR- positive / HER-2 negative, locally advanced or metastatic breast cancer, with one or more PIK3 CA / AKT 1 / PTEN -alterations as detected by an FDA-approved test.

[0183] Embodiment 28. The method of embodiment 22, wherein the breast cancer in the subject is HR- positive / HER-2 negative, locally advanced or metastatic breast cancer, with one or morePIK3 CA / AKT 1 / PTEN -alterations as detected by an FDA-approved test following progression on at least one endocrine-based regimen in the metastatic setting or recurrence on or within 12 months of completing adjuvant therapy.

[0184] Embodiment 29. The method of embodiment 22, wherein the breast cancer is triple negative breast cancer (TNBC).

[0185] Embodiment 30. The method of embodiment 29, wherein the TNBC is selected from the group consisting of basal -like TNBC, mesenchymal TNBC, immunomodulatory TNBC, and luminal androgen receptor TNBC.

[0186] Embodiment 31. The method of embodiment 21, wherein the cancer in the subject is prostate cancer.

[0187] Embodiment 32. The method of embodiment 31, wherein the prostate cancer in the subject is selected from metastatic cancer, non-metastatic cancer, metastatic castration-resistant cancer, metastatic castration-sensitive cancer, localized high risk cancer, recurrent cancer, non-metastatic castration-resistant cancer, non-metastatic castration-sensitive cancer, androgen receptor inhibitor-sensitive cancer, androgen receptor inhibitor-resistant cancer, androgen receptor-dependent cancer, androgen receptor-independent cancer, neuroendocrine cancer (NEPC), metastatic neuroendocrine cancer (NEPC), cancer with small cell features, metastatic cancer with small cell features, and aggressive-variant cancer.

[0188] Embodiment 33. The method of embodiment 32, wherein the prostate cancer in the subject is metastatic cancer.

[0189] Embodiment 34. The method of embodiment 32, wherein the prostate cancer in the subject is non- metastatic cancer.

[0190] Embodiment 35. The method of embodiment 32, wherein the prostate cancer in the subject is metastatic castration-resistant cancer.WSGR Ref. 47134-779.601

[0191] Embodiment 36. The method of embodiment 32, wherein the prostate cancer in the subject is metastatic castration-sensitive cancer.

[0192] Embodiment 37. The method of embodiment 32, wherein the prostate cancer in the subject is localized high risk cancer.

[0193] Embodiment 38. The method of embodiment 32, wherein the prostate cancer in the subject is recurrent cancer.

[0194] Embodiment 39. The method of embodiment 32, wherein the prostate cancer in the subject is non- metastatic castration-resistant cancer.

[0195] Embodiment 40. The method of embodiment 32, wherein the prostate cancer in the subject is non- metastatic castration-sensitive cancer.

[0196] Embodiment 41. The method of embodiment 32, wherein the prostate cancer in the subject is androgen receptor inhibitor-sensitive cancer.

[0197] Embodiment 42. The method of embodiment 32, wherein the prostate cancer in the subject is androgen receptor inhibitor-resistant cancer.

[0198] Embodiment 43. The method of embodiment 32, wherein the prostate cancer in the subject is androgen receptor-dependent cancer.

[0199] Embodiment 44. The method of embodiment 32, wherein the prostate cancer in the subject is androgen receptor-independent cancer.

[0200] Embodiment 45. The method of embodiment 32, wherein the prostate cancer in the subject is neuroendocrine cancer (NEPC).

[0201] Embodiment 46. The method of embodiment 32, wherein the prostate cancer in the subject is metastatic neuroendocrine cancer (NEPC).

[0202] Embodiment 47. The method of embodiment 32, wherein the prostate cancer in the subject is cancer with small cell features.

[0203] Embodiment 48. The method of embodiment 32, wherein the prostate cancer in the subject is metastatic cancer with small cell features.

[0204] Embodiment 49. The method of embodiment 32, wherein the prostate cancer in the subject is aggressive-variant cancer.

[0205] Embodiment 50. The method of any one of embodiments 1 to 49, wherein the AKT inhibitor is selected from ipatasertib (GDC-0068), capivasertib (AZD5363), MK2206, afuresertib (GSK2110183), uprosertib (GSK2141795), perifosine (KRX- 0401), PHT-427 (CS-0223), and Akti-1 / 2.

[0206] Embodiment 51. The method of embodiment 50, wherein the AKT inhibitor is ipatasertib (GDC- 0068).

[0207] Embodiment 52. The method of embodiment 50, wherein the AKT inhibitor is capivasertib (AZD5363).

[0208] Embodiment 53. The method of embodiment 50, wherein the AKT inhibitor is MK2206.WSGR Ref. 47134-779.601

[0209] Embodiment 54. The method of embodiment 50, wherein the AKT inhibitor is afuresertib (GSK2110183).

[0210] Embodiment 55. The method of embodiment 50, wherein the AKT inhibitor is uprosertib (GSK2141795).

[0211] Embodiment 56. The method of embodiment 50, wherein the AKT inhibitor is perifosine (KRX- 0401).

[0212] Embodiment 57. The method of embodiment 50, wherein the AKT inhibitor is PHT-427 (CS- 0223).

[0213] Embodiment 58. The method of embodiment 50, wherein the AKT inhibitor is Akti-1 / 2.

[0214] Embodiment 59. The method of any one of embodiments 1 to 58, wherein the subject is not administered an androgen receptor inhibitor.

[0215] Embodiment 60. The method of any one of embodiments 1 to 59, wherein the subject is administered one or more additional therapeutic agent.

[0216] Embodiment 61. The method of embodiment 60, wherein the one or more additional therapeutic agents is selected from an androgen receptor degrader, a chemotherapeutic agent, a mitotic inhibitors, an antimetabolites, a platinum -based agents, histone deacetylase (HDAC) inhibitors, CD30 -directed antibody-drug conjugates, famesyl transferase inhibitors, SYK inhibitors, JAK inhibitors, PI3K pathway inhibitors, immunomodulatory agents, radiopharmaceuticals, PARP inhibitors, or combinations thereof.

[0217] Embodiment 62. The method of embodiment 61, wherein the one or more additional therapeutic agents is not an androgen receptor inhibitor.

[0218] Embodiment 63. The method of embodiment 61, wherein the one or more additional therapeutic agents is not apalutamide, darolutamide, enzalutamide, or abiraterone.EXAMPLESExample 1: Preparation of crystalline Form 1 of Compound 1

[0219] 150 pL of methanol was added to 50 mg of the free base of Compound 1 and the resulting slurry was stirred at room temperature for one day. The resulting solids were vacuum fdtered and dried under ambient condition overnight to afford Form 1 of Compound 1.Example 2A: Preparation of crystalline Form 2 of Compound 1

[0220] 400 mg of the free base of Compound 1 was dissolved in 1.5 mb of 2-methyltetrahydrofuran at 50 °C, to which was added 1.5 mb n-heptane at about 47 °C, and the resulting mixture was cooled 10 °C.The resulting solids were vacuum filtered and allowed to air dry overnight under ambient conditions to afford Form 2 of Compound 1.Example 2B: Preparation of crystalline Form 2 of Compound 1

[0221] A quantity of the free base of Compound 1 was dissolved in 2-methyltetrahydrofuran (10 volumes) and then distilled to 3 volumes. The temperature of the solution was adjusted to about 25 °C and the resulting slurry was stirred for greater than 30 minutes. To the slurry was added n-heptane (7 volumes)WSGR Ref. 47134-779.601 over 2 hours and the resulting mixture was stirred for greater than 4 hours. The resulting solid was fdtered, the fdter cake was washed with 30% 2 -methyltetrahydrofuran / heptane (2 volumes) and dried in a vacuum oven to provide Form 2 of Compound 1.Example 3A: X-ray powder diffraction (XRPD) analysis of Form 1 and Form 2 of Compound 1

[0222] XRPD analyses of crystalline polymorphic forms of Compound 1 were performed using Panalytical X’pert3X-ray powder diffractometer. Samples were spread on the middle of a zerobackground Si holder. The 2-theta position was calibrated against a Panalytical Si reference standard disc.The parameters used for the analyses are set forth in Table 1.Table 1

[0223] Polymorphic Form 1 of Compound 1 was analyzed by XRPD as set forth above and exhibited the peaks set forth in Table 2. The error associated with each °2 -theta position was determined to be ± 0.2° theta.Table 2

[0224] Polymorphic Form 2 of Compound 1 was analyzed by XRPD as set forth above and exhibited the peaks set forth in Table 3. The error associated with each °2 -theta position was determined to be ± 0.2 °- theta.Table 3WSGR Ref. 47134-779.601Example 3B: Thermal gravimetric analyses and differential scanning calorimetry analyses of Form 1 and Form 2 of Compound 1

[0225] Thermal gravimetric analysis (TGA) data were collected using TA Discovery TGA 550 TGA from TA Instruments, and differential scanning calorimetry (DSC) analyses were performed using a TA Q2000 DSC from TA Instruments using the parameters set forth in Table 4.Table 4

[0226] A thermal gravimetric analysis (TGA) of a sample of Form 1 of Compound 1 , when conducted under the conditions set forth in Table 4, exhibited a weight loss of about 1% upon heating the sample from room temperature to about the onset of melting (about 207 °C). A differential scanning calorimetry (DSC) analysis of Form 1 of Compound 1, when conducted under the conditions set forth in Table 4, exhibited peaks at between about 170 °C and 172 °C, and between about 207 °C and 208 °C.

[0227] A thermal gravimetric analysis (TGA) of a sample of Form 2 of Compound 1 , when conducted under the conditions set forth in Table 4, exhibited a weight loss of about 2% upon heating the sample from room temperature to about the onset of melting (about 204 °C). A differential scanning calorimetry (DSC) analysis of Form 2 of Compound 1, when conducted under the conditions set forth in Table 4, exhibited a peak between about 203 °C and 204 °C.Example 3C: Preparation of crystalline Form 2 of Compound 1

[0228] A reactor was evacuated and charged with nitrogen to atmospheric pressure. The reactor was then charged with a solution of Compound 1 (approximately 2.41 kg as determined by solution assay using HPLC) in 2 -methyltetrahydrofuran (2-MeTHF, 36 kg, 15 volumes) and the batch was concentrated to a batch volume of about 5 L (about 2 volumes) via distillation under reduced pressure. The resulting solution was adjusted to about 25 °C and then n-heptane (0.4 kg, 0.2 volumes) was added in portions over a period of about 3 hours. The resulting solution was then seeded with Compound 1 Form 2 (9 g, 0.4 wt%), the resulting mixture was stirred for about 1.3 hours, and then additional n-heptane (24 kg, 10WSGR Ref. 47134-779.601 volumes) was added over about 6 hours. The resulting slurry was stirred for about 4.25 hours at 25 °C and then filtered. The reactor was then rinsed with n-heptane (5.8 kg, 2.5 V), and this mixture was rinsed forward to the filter cake, which was deliquored and the solids were dried under reduced pressure at 40 °C and 50 °C for 19 hours to provide 2.48 kg of Form 2 of Compound 1.Example 4: Cell viability assays in C4-2 and 22RV1 cells using a combination of Compound 1 and an AKT inhibitor

[0229] The effect of test articles was measured based on viability of C4-2 and 22RVlcells by CellTiter- Glo® (CTG) assays 14 days post compound treatment.

[0230] Materials: (a) cell lines: C4-2 and 22Rvl were obtained from American Type Culture Collection (ATCC); (b) culture media for both cell lines: RPMI + 10% FBS + 2 mM L-Glut + 0.5 pg / m penicillinstreptomycin; RPMI1640 with phenol red (Coming, Cat#: 15-040-CV); fetal bovine serum (FBS) (Omega Scientific, Cat#: FB-11); L-glutamine (Coming, Cat#: 25-005-CI); penicillin-streptomycin (Gibco, Cat#: 15140122); (c) assay media: Same as culture medium; (d) CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Cat#: G9243); (e) DMSO (Sigma, Cat#: D2660); (f) Compound 1 (ORIC Pharmaceuticals, Inc.; (g) capivasertib (Selleckchem, Cat#: S8019).

[0231] Equipment: (a) Bravo Liquid Handler (Agilent, Cat#: G5523BA); (b) 96-well Non-Sterile Polypropylene V Bottom Plates (Coming, Cat#: 3363); (c) 384-well Sterile White with Flat Clear Bottom Plates (Greiner, Cat#: 781098); (d) TopSeal-A Plus plate seal (PerkinElmer, Cat#: 6050185)

[0232] Cell handling: C4-2 and 22RV1 cells were cultured in complete culture media at exponential growth phase for at least one week prior to assay. The different cell suspensions were counted and seeded into 384-well Sterile White Flat Clear Bottom Plates using a Bravo liquid handler at a concentration of 500 cells in 56 pL of media per well.

[0233] Compound preparation, dilution and dispensing: To prepare the compound dilutions from 10 mM stock solutions, dilution plates were prepared using the Bravo liquid handler by performing 10-point 1 / 3- fold serial dilutions in DMSO (for capivasertib) or 6-point 1 / 3-fold serial dilutions in DMSO (for Compound 1) to a final concentration of 500-fold the initial concentration in the final assay plate. For simultaneous dosing, compound dilutions in media were added to cells by dispensing 2 pL into cells plated the same day in 56 pL of culture media. Plates were placed in a 37 °C, 5% CO2 incubator.Following 7 days of incubation the cell culture media and inhibitors were replenished and incubated for an additional 4 days. For sequential dosing, Compound 1 dilutions in media were added to cells by dispensing 2 pL into cells plated the same day in 58 pL of culture media. Plates were placed in a 37 °C, 5% CO2 incubator. Following 7 days of incubation the cell culture media and inhibitors were replenished with compound dilutions containing both Compound 1 and capivasertib and incubated for an additional 4 days. Prior to performing the viability assay, plates were allowed to equilibrate at room temperature before the addition of 1: 1 ratio of CTG 2.0 using the Bravo liquid handler. The plates were sealed with a PerkinElmer TopSeal -A Plus plate seal and incubated at room temperature for 15 minutes before readingWSGR Ref. 47134-779.601 the luminescence on a plate reader (Clariostar-plus). The luminescence is a direct readout of the presence of ATP in the cells for measure of cell viability.

[0234] Data analysis: Data from the plate reader was transferred to a Microsoft Excel file and entered into GraphPad Prism. Curves were fitted with a four-parameter model and % Inhibition calculated by GraphPad Prism. DMSO wells were used to define the upper limit (0% inhibition). The degree of combination synergy, neutrality, or antagonism, was quantified by comparing the observed drug combination response against the expected response, calculated using a reference model that assumes no interaction between drugs using the SynergyFinder website (lanevski 2022). These models quantify the degree of synergy with different algorithms, either as the excess over the maximum single drug response (HSA), multiplicative effect of single drugs as if they acted independently (Bliss), or expected response corresponding to an additive effect as if the single drugs were the same compound (Loewe).

[0235] Results: The synergy scores measured for the combination of Compound 1 and capivasertib are set forth in Table 5.Table 5

Claims

WSGR Ref. 47134-779.601CLAIMSWHAT IS CLAIMED is:

1. A method of treating cancer in a subject, wherein the cancer in the subject has been determined to comprise one or more PIK3CA / AKTl / PTEN-alterations, comprising administering to the subject(a) Compound(Compound 1), or a pharmaceutically acceptable salt thereof, and (b) an AKT inhibitor.

2. The method of claim 1, wherein the cancer in the subject has been determined to comprise one or more PIK3CA alterations.

3. The method of claim 1, wherein the cancer in the subject has been determined to comprise one or more AKT1 alterations.

4. The method of claim 1, wherein the cancer in the subject has been determined to comprise one or more PTEN alterations.

5. The method of claim 4, wherein the cancer in the subject has been determined to be PTEN-null.

6. The method of any one of claims 1 to 5, wherein the subject has received prior administration of one or more chemotherapeutic agents prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

7. The method of any one of claims 1 to 5, wherein the subject has not been administered a chemotherapeutic agent prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

8. The method of any one of claims 1 to 5, wherein the subject has been administered at least one endocrine -based therapy prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.

9. The method of claim 8, wherein the cancer in the subject has progressed on the at least one endocrine -based therapy prior to the administration to the subject of Compound 1, or a pharmaceutically acceptable salt thereof, and the AKT inhibitor.WSGR Ref. 47134-779.60110. The method of any one of claims 1 to 9, wherein the cancer in the subject is selected from diffused large B cell lymphoma (DLBCL), follicular lymphoma, other lymphomas, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, cancer, breast cancer, bile duct and gallbladder cancers, bladder carcinoma, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal carcinoma, ovarian cancer, pancreatic cancer, renal cell carcinoma, prostate cancer, rectal cancer, thyroid cancers, parathyroid tumors, uterine tumors, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), Kaposi sarcoma, synovial sarcoma, osteosarcoma and Ewing's sarcoma.

11. The method of claim 10, wherein the cancer in the subject is breast cancer.

12. The method of claim 11, wherein the breast cancer in the subject is selected from HR-positive breast carcinoma, HR-negative breast carcinoma, primary breast ductal carcinoma, mammary adenocarcinoma, HR-positive mammary ductal carcinoma, HR- negative mammary ductal carcinoma, HER2 -positive mammary ductal carcinoma, HER2 -positive breast cancer, luminal breast cancer, triple negative breast cancer (TNBC), ER-positive breast cancer, ER-negative breast cancer, PR-positive breast cancer, PR-negative breast cancer, HR-positive / HER-2 negative breast cancer, HR-negative / HER-2 negative breast cancer, HR-positive / HER-2 positive breast cancer, and HR-negative / HER-2 positive breast cancer, and unclassified breast cancer.

13. The method of claim 10, wherein the cancer in the subject is prostate cancer.

14. The method of claim 13, wherein the prostate cancer in the subject is selected from metastatic cancer, non-metastatic cancer, metastatic castration-resistant cancer, metastatic castrationsensitive cancer, localized high risk cancer, recurrent cancer, non-metastatic castration-resistant cancer, non-metastatic castration-sensitive cancer, androgen receptor inhibitor-sensitive cancer, androgen receptor inhibitor-resistant cancer, androgen receptor-dependent cancer, androgen receptor-independent cancer, neuroendocrine cancer (NEPC), metastatic neuroendocrine cancer (NEPC), cancer with small cell features, metastatic cancer with small cell features, and aggressive-variant cancer.

15. The method of any one of claims 1 to 14, wherein the AKT inhibitor is selected from ipatasertib (GDC-0068), capivasertib (AZD5363), MK2206, afuresertib (GSK2110183), uprosertib (GSK2141795), perifosine (KRX- 0401), PHT-427 (CS-0223), and Akti-1 / 2.

Images