Pharmaceutical forms of CD73 inhibitors

Abstract

The present disclosure relates to pharmaceutical forms of a CD73 inhibitor, polymorphic forms of the pharmaceutical forms, pharmaceutical compositions comprising the pharmaceutical forms, and methods of treating a subject having cancer by administering one of the pharmaceutical forms or a pharmaceutical composition comprising one of the disclosed pharmaceutical forms.

Description

[0001] Cross-reference

[0002] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 375,580, filed Sep. 14, 2022, and U.S. Provisional Application Serial No. 63 / 497,323, filed Apr. 20, 2023, which are hereby incorporated by reference in their entireties. BACKGROUND OF THE INVENTION

[0003] CD73 is a glycosylphosphatidylinositol (GPI)-anchored cell surface protein that catalyzes the hydrolysis of AMP to adenosine and acts in concert with CD39, which converts ATP to AMP. The resulting adenosine serves as a signaling molecule that activates P1 receptors expressed on the cell surfaces in many different tissues. Four G protein-coupled P1 or adenosine receptors have been cloned and designated A1, A2A, A2B, and A3. Adenosine affects a wide range of physiological processes, including neural function, vascular perfusion, and immune responses. In this context, this metabolite modulates CNS, cardiovascular, and immune system functions, to name just a few.

[0004] Increasing evidence indicates that the interaction between tumor cells and their microenvironment is essential for tumorigenesis. The purinergic signaling pathway, in which CD73 plays a key role, has emerged as an important player in cancer progression. In recent years, it has become clear that adenosine is one of the most important immunosuppressive regulatory molecules in the tumor microenvironment and contributes to immune escape and tumor progression.

[0005] CD73 is a key protein molecule in cancer development. CD73 has been found to be overexpressed in many cancer cell lines and tumor types, including, for example, breast cancer, colorectal cancer, ovarian cancer, gastric cancer, gallbladder cancer, and cancers associated with poor prognosis.

[0006] The expression of CD73 in tumors is regulated by multiple mechanisms. In breast cancer, CD73 expression is negatively regulated by the estrogen receptor (ER). Thus, CD73 is highly expressed in patients with ER-negative breast cancer. Hypoxia-inducible factor-1α (HIF-1α) has also been shown to regulate CD73 transcription. Additionally, inflammatory cytokines such as IFN-γ affect CD73 levels. CD73 expression is also epigenetically regulated by CpG island methylation in cell lines and clinical tumor samples.

[0007] In addition to being a prognostic biomarker in cancer patients, the overexpression of CD73 has also been found to be functionally associated with therapy resistance. Elevated levels of CD73 were initially associated with resistance to multiple chemotherapeutic agents, including vincristine and doxorubicin.

[0008] CD73 has also been shown to be involved in immunotherapy resistance. This ectonucleotidase participates in the process of tumor immune escape by inhibiting the activation, clonal expansion, and homing of tumor-specific T cells (especially T helper cells and cytotoxic T cells); impairing tumor cell killing achieved by cytolytic effector T lymphocytes; driving the inhibitory capacity of Tregs and Th17 cells by generating adenosine around cells; enhancing the transformation of type 1 macrophages into pro-tumor type 2 macrophages; and promoting the accumulation of MDSCs.

[0009] Small molecule inhibitors and monoclonal antibodies targeting CD73 have shown anti-tumor activity in multiple immunocompetent mouse tumor models but not in immunodeficient mouse tumor models. Collectively, these studies suggest that the activity of anti-CD73 therapy depends on its ability to elicit an immune response in vivo.

[0010] Antibodies blocking PD-1, PD-L1, and CTLA-4 have shown impressive objective responses in cancer patients. New data demonstrate that in several mouse tumor models, anti-CD73 mAb significantly enhances the activity of both anti-CTLA-4 mAb and anti-PD-1 mAb. In addition to checkpoint blockade, CD73-mediated adenosine production may also contribute to resistance to additional immunotherapy modalities including CAR-T cells and cancer vaccines.

[0011] Interfering with CD73 activity represents a strategy to re-sensitize tumors to therapy. Based on the association between CD73 and therapy resistance, combining anti-CD73 treatment with chemotherapy or immunotherapy is an effective way to enhance their activity in cancer patients with high CD73 levels. In some cases, CD73 expression serves as a biomarker to identify patients who may benefit from anti-CD73 combination therapy.

[0012] In some cases, the CD39 / CD73 couple shifts ATP-driven pro-inflammatory cell activity towards an adenosine-mediated anti-inflammatory state. Many studies have shown that the activity of the CD39 / CD73 axis changes during infections induced by various microorganisms. An increase in CD73 expression has also been observed in the brains of mice infected with Toxoplasma gondii, which promotes the parasite life cycle by generating adenosine. Thus, pharmacological blockade of CD73 is a promising therapeutic approach for treating human toxoplasmosis.

[0013] Enhanced expression and activity of CD39 and CD73 have been observed in endothelial cells infected with cytomegalovirus (CMV). Increased local adenosine production associated with the upregulation of ectonucleotidases creates an immunosuppressive and antithrombotic microenvironment that promotes viral entry into target cells.

[0014] In some cases, inhibitors of CD73 are used as antiviral agents by driving down adenosine production. Elevated expression / activity of CD39 and CD73 on lymphocytes of individuals infected with human immunodeficiency virus (HIV) indicates that ectonucleotidases have a role in immune dysfunction associated with this disease. Indeed, an increased proportion of CD39-expressing Tregs and a positive association between CD39 expression on Tregs and disease progression have been observed in different cohorts of HIV-infected patients. It has also been shown that HIV-positive patients have a higher number of CD39+ Tregs and that their Teffs exhibit increased sensitivity to the inhibitory effects of adenosine in vitro, which is associated with elevated expression of the immunosuppressive A2A receptor.

[0015] In the central nervous system, adenosine plays a key role in controlling numerous neurological functions. By activating P1 receptors, adenosine is involved in a variety of physiological and pathological processes, such as the regulation of sleep, general arousal state and activity, local neuronal excitability, and the coupling of cerebral blood flow and energy requirements. In some cases, manipulation of adenosine production by CD73 inhibitors can be used to treat neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease, as well as psychiatric disorders such as schizophrenia and autism.

[0016] Accordingly, there is a need to develop CD73 inhibitors and pharmaceutical compositions comprising such CD73 inhibitors that can be advantageously used to treat cancer and infections in subjects in need thereof. SUMMARY OF THE INVENTION

[0017] There is provided herein a pharmaceutically acceptable form of Compound 1:

[0018]

[0019] The pharmaceutically acceptable forms are selected from the gentisate form and the succinate form. The present disclosure also provides polymorphic forms of the gentisate form and the succinate form of Compound 1. In some embodiments, the gentisate form comprises a gentisate salt. In some embodiments, the gentisate form comprises a gentisate cocrystal. In some embodiments, the gentisate form comprises a salt and a cocrystal. In some embodiments, the succinate form comprises a succinate salt. In some embodiments, the succinate form comprises a succinate cocrystal. In some embodiments, the succinate form comprises a salt and a cocrystal. The present disclosure further provides a pharmaceutical composition comprising a pharmaceutically acceptable form of Compound 1 in an amount, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form. Some embodiments disclosed herein provide a method of inhibiting CD73 activity in a cell, the method comprising contacting CD73 in the cell with an effective amount of a pharmaceutically acceptable form of Compound 1 as disclosed herein. The present disclosure further provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1 as disclosed herein. In some embodiments, the cancer is lung cancer, melanoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, prostate cancer, kidney cancer, multiple myeloma, or lymphoma. In other embodiments, the cancer expresses CD73. In still further embodiments, a method of treating cancer in a subject in need thereof is provided, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1 as disclosed herein, and one or more second therapeutic agents. Detailed Description

[0020] Unless the context clearly dictates otherwise, as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents. Thus, for example, reference to "an agent" includes plural such agents 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, melting point or chemical properties such as chemical formula, all combinations and subcombinations of the ranges and specific embodiments therein are intended to be included. The term "about" when referring to a numerical value or numerical range means that the recited numerical value or numerical range is an approximation within experimental variability (or within statistical experimental error), and thus, in some instances, the numerical value or numerical range will vary between 1% and 15% of the stated numerical value or numerical range. The term "comprising" (and related terms such as "includes" or "having") is not intended to exclude the case of "consisting of the described features" or "consisting essentially of the described features" in other certain embodiments described herein, such as embodiments of any substance composition, composition, method, or process, etc.

[0021] As used in the specification and the appended claims, unless specified to the contrary, the following terms have the meanings indicated below.

[0022] "Administer" when used in connection with a therapeutic means is to deliver a therapeutic agent systemically or locally, such as directly into or onto a target tissue, or to administer a therapeutic agent to a subject, whereby the therapeutic agent actively affects the tissue to which it is targeted. Thus, as used herein, the term "administer" when used in connection with the compositions described herein can include, but is not limited to, delivering the composition into or onto a target tissue; delivering the composition systemically to a subject, such as by oral administration, such that the therapeutic agent reaches the target tissue or target cell. Administration of the "composition" can be accomplished by injection, topical application, and oral administration or by other methods alone or in combination with other known techniques.

[0023] The term "amorphous" as used herein refers to a solid composition that has no measurable long-range order in the positions of its molecules, as measured by analytical techniques known to those of ordinary skill in the art (such as X-ray powder diffraction (XRPD)).

[0024] The terms "crystalline" and "crystallinity" refer to a solid composition that has a degree of long-range order in the positions of its molecules, as measured by analytical techniques known to those of ordinary skill in the art (such as X-ray powder diffraction (XRPD)).

[0025] The term "differential scanning calorimetry" as used herein refers to the thermal analysis method described in USP <891>.

[0026] The term "gentisate" as used herein refers to a salt formed between Compound 1 and gentisic acid. The term "gentisic acid" as used herein refers to the compound having Chemical Abstracts Registry Number 490-79-9, chemical name 2,5-dihydroxybenzoic acid, and the following chemical structure:

[0027]

[0028] The term "gentisate" as used herein refers to the form in which Compound 1 associates with gentisic acid. As contemplated herein, the gentisate can be (a) a salt form comprising Compound 1 and gentisic acid, (b) a co-crystal comprising Compound 1 and gentisic acid, or (c) a mixture of a salt of Compound 1 and gentisic acid and a co-crystal of Compound 1 and gentisic acid.

[0029] As used herein, the term "cocrystal" refers to a crystalline material that contains two or more different molecules (one of which is Compound 1) in a defined stoichiometry that are associated by non-ionic and non-covalent bonds within the same crystal lattice. In one embodiment, a cocrystal comprising Compound 1 and gentisic acid is provided. In another embodiment, a cocrystal comprising Compound 1 and succinic acid is provided.

[0030] The term "pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not harmful to its recipient.

[0031] The term "pharmaceutical composition" shall mean a composition comprising one or more active ingredients, whereby the composition is suitable for conducting studies in mammals (such as but not limited to humans) for a specific beneficial effect. Those of ordinary skill in the art will understand and appreciate the techniques applicable to determining whether an active ingredient has the desired beneficial effect based on the needs of the skilled person.

[0032] As used herein, the term "substantially amorphous" refers to a composition having little or no long-range order in the positions of its molecules. For example, a substantially amorphous material has a crystallinity of less than about 15% (e.g., less than about 10% crystallinity or less than about 5% crystallinity). As contemplated herein, the term substantially amorphous includes compositions containing immeasurable crystalline material, as measured by analytical techniques known to those of ordinary skill in the art (such as X-ray powder diffraction (XRPD)).

[0033] As used herein, the term "succinate" refers to a salt formed between Compound 1 and succinic acid. As used herein, the term "succinic acid" refers to the compound having Chemical Abstracts Registry Number 110-15-6, chemical name 1,2-ethanedicarboxylic acid and 1,4-butanedioic acid and the following chemical structure:

[0034]

[0035] As used herein, the term "succinate" refers to the form in which Compound 1 is associated with succinic acid. As contemplated herein, the succinate can be (a) a salt form comprising Compound 1 and succinic acid, (b) a cocrystal comprising Compound 1 and succinic acid, or (c) a mixture of a salt comprising Compound 1 and succinic acid and a cocrystal comprising Compound 1 and succinic acid.

[0036] As used herein, the term "therapeutic agent" refers to an agent for treating, combating, alleviating, preventing or improving an unwanted condition or disease in an object.

[0037] As used herein, "therapeutically effective amount" or "effective amount" refers to the amount of an active compound or agent that elicits a biological or medical response sought by a researcher, veterinarian, physician, or other clinician in a tissue, system, animal, individual, or human, and includes one or more of the following: (1) preventing a disease; e.g., preventing a disease, condition, or disorder in an individual who may be predisposed to the disease, condition, or disorder but has not yet experienced or exhibited the pathology or symptoms of the disease, (2) inhibiting a disease; e.g., inhibiting a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (i.e., preventing further development of the pathology and / or symptoms), and (3) alleviating a disease; e.g., alleviating a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (i.e., reversing the pathology and / or symptoms).

[0038] As used herein, the term "treat", "treated", "treatment", or "treating" refers to therapeutic treatment in some embodiments and prophylactic or preventive measures in other embodiments, where the goal is to prevent or slow down (mitigate) an undesired physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical outcome. For the purposes described herein, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms; reduction in the degree of a condition, disorder, or disease; stabilization of a condition, disorder, or disease state (i.e., not getting worse); delay in the onset or slowing of the progression of a condition, disorder, or disease; alleviation of a condition, disorder, or disease state; and remission (whether detectable or undetectable, partial or complete) or enhancement or improvement of a condition, disorder, or disease. Treatment includes eliciting a clinically significant response without undue side effects. Treatment also includes prolonging survival as compared to the expected survival if treatment is not received. The prophylactic benefits of treatment include preventing a condition, delaying the progression of a condition, stabilizing a condition, or reducing the likelihood of a condition occurring. As used herein, "treat" in some embodiments includes prevention.

[0039] As used herein, the term "X-ray powder diffraction (XRPD)" refers to a technique for characterizing the crystallinity or partial crystallinity of a solid by using powder X-ray diffraction, as set forth in USP <941>.

[0040] The present disclosure provides pharmaceutically acceptable forms of Compound 1:

[0041]

[0042] wherein the pharmaceutically acceptable forms are selected from the gentisate form and the succinate form.

[0043] In some embodiments, a pharmaceutically acceptable form of Compound 1 is provided, wherein the form is the gentisate form. In some embodiments, the gentisate form comprises a gentisate salt. In some embodiments, the gentisate form comprises a gentisate cocrystal. In some embodiments, the gentisate form comprises a salt and a cocrystal. In further embodiments, a gentisate form of Compound 1 is provided, wherein the molar ratio between Compound 1 and gentisic acid is about 1:1. In further embodiments, a gentisate form of Compound 1 is provided, wherein the molar ratio between Compound 1 and gentisic acid is about 2:1. In further embodiments, a gentisate form of Compound 1 is provided, wherein the molar ratio between Compound 1 and gentisic acid is about 1:2. In other embodiments, a gentisate form of Compound 1 is provided, wherein the gentisate form is a hydrate. In some embodiments, the hydrate of the gentisate form of Compound 1 is selected from the hemihydrate, monohydrate, and dihydrate forms. In some embodiments, the hydrate is the hemihydrate. In some embodiments, the hydrate is the monohydrate. In some embodiments, the hydrate is the dihydrate.

[0044] In further embodiments, a gentisate form of Compound 1 is provided, wherein the form is a solid. In some embodiments, such solid is crystalline. In some embodiments, such crystalline solid exhibits a peak at 9.25 ± 0.2° 2-θ in an X-ray powder diffraction (XRPD) pattern. In still further embodiments, the crystalline solid exhibits further peaks at 6.97 ± 0.2° 2-θ, 20.53 ± 0.2° 2-θ, and 26.08 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In still further embodiments, the crystalline solid exhibits further peaks at 14.61 ± 0.2° 2-θ and 18.89 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0045] In a further embodiment, there is provided a gentisate form of Compound 1, wherein said form is a crystalline solid and exhibits a peak at 9.3 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In yet a further embodiment, the crystalline solid exhibits a further peak at 7.1 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In yet a further embodiment, the crystalline solid exhibits a further peak at 20.1 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In yet a further embodiment, the crystalline solid exhibits a further peak at 19.0 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In yet a further embodiment, the crystalline solid exhibits a further peak at 26.2 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In yet a further embodiment, the crystalline solid exhibits further peaks at 4.8 ± 0.2° 2-θ, 10.5 ± 0.2° 2-θ, and 14.8 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0046] In other embodiments, there is provided a crystalline solid of a gentisate form of Compound 1, wherein said crystalline form comprises a peak in the differential scanning calorimetry (DSC) thermogram from about 150 °C to about 170 °C. In yet a further embodiment, the crystalline solid comprises a peak in the differential scanning calorimetry (DSC) thermogram from about 150 °C to about 165 °C. In yet a further embodiment, the crystalline solid comprises a peak in the differential scanning calorimetry (DSC) thermogram from about 161 °C to about 162 °C.

[0047] In other embodiments, there is provided a crystalline solid of a gentisate form of Compound 1, wherein said crystalline solid exhibits a mass loss between about 1% and about 5% in thermogravimetric analysis when heated from about 31 °C to about 150 °C. In yet a further embodiment, there is provided a crystalline solid of a gentisate form of Compound 1, wherein said crystalline solid exhibits a mass loss between about 3% and about 5% in thermogravimetric analysis when heated from about 31 °C to about 150 °C. In yet a further embodiment, there is provided a crystalline solid of a gentisate form of Compound 1, wherein said crystalline solid exhibits a mass loss of about 5% in thermogravimetric analysis when heated from about 31 °C to about 150 °C.

[0048] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 5 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C. In a further embodiment, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 10 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C. In a further embodiment, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 15 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C. In a further embodiment, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 20 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C. In a further embodiment, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 25 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C.

[0049] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility between about 10 mg / mL and about 30 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility between about 15 mg / mL and about 30 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility between about 20 mg / mL and about 30 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility between about 25 mg / mL and about 30 mg / mL in an aqueous solution having a pH of 1.7 and a temperature of 37 °C.

[0050] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 5 mg / mL in an aqueous solution having a pH of 2.5 and a temperature of 37°C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 7.5 mg / mL in an aqueous solution having a pH of 2.5 and a temperature of 37°C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 10 mg / mL in an aqueous solution having a pH of 2.5 and a temperature of 37°C.

[0051] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 1 mg / mL in an aqueous solution having a pH of 4.4 and a temperature of 37°C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 2.5 mg / mL in an aqueous solution having a pH of 4.4 and a temperature of 37°C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 5 mg / mL in an aqueous solution having a pH of 4.4 and a temperature of 37°C. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid exhibits a solubility of at least 7.5 mg / mL in an aqueous solution having a pH of 4.4 and a temperature of 37°C.

[0052] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 5 °C for at least 7 days, the crystalline solid exhibits less than about 10% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 5 °C for at least 7 days, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 5 °C for at least one month, the crystalline solid exhibits less than about 10% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 5 °C for at least one month, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 5 °C for at least 3 months, the crystalline solid exhibits less than about 10% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 5 °C for at least 3 months, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% degradation of the total amount of the gentisate form of Compound 1.

[0053] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 25 °C and 60% relative humidity for at least 7 days, the crystalline solid exhibits less than about 10% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 25 °C and 60% relative humidity for at least 7 days, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 25 °C and 60% relative humidity for at least one month, the crystalline solid exhibits less than about 10% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 25 °C and 60% relative humidity for at least one month, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 25 °C and 60% relative humidity for at least 3 months, the crystalline solid exhibits less than about 10% degradation of the total amount of the gentisate form of Compound 1. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 25 °C and 60% relative humidity for at least 3 months, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% of the total amount of the gentisate form of Compound 1.

[0054] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 40 °C and 75% relative humidity for at least 7 days, the crystalline solid exhibits less than about 10% degradation. In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 40 °C and 75% relative humidity for at least 7 days, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% of the total amount of the gentisate form of Compound 1.

[0055] In other embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 60 °C for one or more days, the crystalline solid exhibits less than about 10% degradation of the total amount of the gentisate form of Compound 1. In still further embodiments, a crystalline solid of the gentisate form of Compound 1 is provided, wherein when the pharmaceutically acceptable form is stored at 60 °C for one or more days, the crystalline solid exhibits less than about 9%, or less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5% degradation of the total amount of the gentisate form of Compound 1.

[0056] Also provided herein is a crystalline solid of the gentisate form of Compound 1, wherein the crystalline solid exhibits a peak at 8.26 ± 0.2° 2-θ in an X-ray powder diffraction (XRPD) pattern. Further provided herein is a crystalline solid of the gentisate form of Compound 1, wherein the crystalline solid further exhibits a peak at 26.43 ± 0.2° 2-θ in an X-ray powder diffraction (XRPD) pattern. In another embodiment, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid further exhibits peaks at 15.81 ± 0.2° 2-θ and 15.40 ± 0.2° 2-θ in an X-ray powder diffraction (XRPD) pattern. In another embodiment, a crystalline solid of the gentisate form of Compound 1 is provided, wherein the crystalline solid further exhibits peaks at 14.94 ± 0.2° 2-θ and 20.44 ± 0.2° 2-θ in an X-ray powder diffraction (XRPD) pattern.

[0057] The present invention also provides a pharmaceutically acceptable form of Compound 1, wherein the form is the succinate form. In some embodiments, the succinate form comprises succinate. In some embodiments, the succinate form comprises a succinate cocrystal. In a further embodiment, a succinate form of Compound 1 is provided, wherein the molar ratio between Compound 1 and succinic acid is about 1:1. In a further embodiment, a succinate form of Compound 1 is provided, wherein the molar ratio between Compound 1 and succinic acid is about 2:1. In a further embodiment, a succinate form of Compound 1 is provided, wherein the molar ratio between Compound 1 and succinic acid is about 1:2. In other embodiments, such succinate forms of Compound 1 are provided, wherein the succinate form is a hydrate. In some embodiments, the hydrate of the succinate form of Compound 1 is selected from the hemihydrate, monohydrate, and dihydrate forms. In some embodiments, the hydrate is the hemihydrate. In some embodiments, the hydrate is the monohydrate. In some embodiments, the hydrate is the dihydrate.

[0058] The present invention further provides a pharmaceutical composition, which comprises an amount of a pharmaceutically acceptable form of Compound 1 and one or more pharmaceutically acceptable excipients, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein. In some embodiments, the gentisate form comprises gentisate. In some embodiments, the gentisate form comprises a gentisate cocrystal. In some embodiments, the gentisate form comprises a salt and a cocrystal. In some embodiments, the succinate form comprises succinate. In some embodiments, the succinate form comprises a succinate cocrystal. In some embodiments, the succinate form comprises a salt and a cocrystal.

[0059] The present invention also provides a method for inhibiting CD73 activity in cells, the method comprising contacting CD73 in the cells with an effective amount of a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein.

[0060] The present invention further provides a method for treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein. In other embodiments, there is provided a method for treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein. In some embodiments, the cancer is lung cancer, melanoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, prostate cancer, kidney cancer, multiple myeloma or lymphoma. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is melanoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is gallbladder cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the prostate cancer is castration-resistant prostate, metastatic prostate cancer, metastatic castration-resistant prostate cancer or castration-sensitive prostate cancer. In some embodiments, the cancer is kidney cancer. In some embodiments, the cancer is multiple myeloma. In some embodiments, the multiple myeloma is light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma or immunoglobulin E (IgE) myeloma. In some embodiments, the cancer is lymphoma.

[0061] The present disclosure further provides a method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate and succinate forms disclosed herein, and wherein the cancer expresses CD73. In some embodiments, CD73 is upregulated in the cancer to be treated. In some embodiments, the cancer that expresses CD73 or in which CD73 is upregulated is lung cancer, melanoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, prostate cancer, kidney cancer, multiple myeloma, or lymphoma. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is melanoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is gallbladder cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the prostate cancer is castration-resistant prostate, metastatic prostate cancer, metastatic castration-resistant prostate cancer, or castration-sensitive prostate cancer. In some embodiments, the cancer is kidney cancer. In some embodiments, the cancer is multiple myeloma. In some embodiments, the multiple myeloma is light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma, or immunoglobulin E (IgE) myeloma. In some embodiments, the cancer is lymphoma.

[0062] The present disclosure further provides a method of treating cancer in a subject, the method comprising administering to the subject (a) a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate and succinate forms disclosed herein, and (b) one or more second therapeutic agents. In other embodiments, provided is a method of treating cancer in a subject, the method comprising administering to the subject (a) a therapeutically effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate and succinate forms disclosed herein, and (b) one or more second therapeutic agents. In some embodiments, the second therapeutic agent is a chemotherapeutic agent or an immunotherapeutic agent.

[0063] In a further embodiment, the cancer of the subject is multiple myeloma, and the second therapeutic agent is selected from chemotherapy, corticosteroids, immunomodulators, proteasome inhibitors, histone deacetylase (HDAC) inhibitors, monoclonal antibodies against CD38, monoclonal antibodies against SLAMF7, antibody-drug conjugates, and nuclear export inhibitors. In some embodiments, the second therapeutic agent is chemotherapy selected from cyclophosphamide, etoposide (VP-16), doxorubicin, liposomal doxorubicin, melphalan, melphalan flufenamide (melflufen), and bendamustine. In other embodiments, the second therapeutic agent is selected from corticosteroids, including but not limited to dexamethasone and prednisone. In other embodiments, the second therapeutic agent is selected from immunomodulators, including but not limited to thalidomide, lenalidomide, and pomalidomide. In a further embodiment, the second therapeutic agent is selected from proteasome inhibitors, including but not limited to bortezomib, carfilzomib, and ixazomib. In other embodiments, the second therapeutic agent is selected from histone deacetylase (HDAC) inhibitors, including but not limited to Panobinostat. In still further embodiments, the second therapeutic agent is selected from monoclonal antibodies against CD38, including but not limited to daratumumab and isatuximab. In some embodiments, the second therapeutic agent is selected from antibodies against SLAMF7, including but not limited to elotuzumab. In some embodiments, the second therapeutic agent is selected from antibody-drug conjugates, including but not limited to belantamab mafodotin. In some embodiments, the second therapeutic agent is selected from nuclear export inhibitors, including but not limited to selinexor.

[0064] In other embodiments, methods of treating multiple myeloma in a subject are provided, the methods comprising administering to the subject (a) a therapeutically effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein, and (b) one or more second therapeutic agents, wherein the one or more second therapeutic agents include, but are not limited to, combinations of the following agents: lenalidomide (or pomalidomide or thalidomide) and dexamethasone; carfilzomib (or ixazomib or bortezomib), lenalidomide and dexamethasone; bortezomib (or carfilzomib), cyclophosphamide and dexamethasone; elotuzumab (or daratumumab), lenalidomide and dexamethasone; bortezomib, liposomal doxorubicin and dexamethasone; panobinostat, bortezomib and dexamethasone; elotuzumab, bortezomib and dexamethasone; melphalan and prednisone (MP), with or without thalidomide or bortezomib; vincristine, doxorubicin and dexamethasone (referred to as VAD); dexamethasone, cyclophosphamide, etoposide and cisplatin (referred to as DCEP); dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide and etoposide (referred to as DT-PACE), with or without bortezomib; and selinexor, bortezomib and dexamethasone.

[0065] Also provided herein are methods of treating an infection in a subject, the methods comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein. In some embodiments, the infection is a viral infection. In other embodiments, the infection is a parasitic infection.

[0066] Also provided herein are methods of treating a neurodegenerative disease in a subject, the methods comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein. Also provided are methods of treating a neurodegenerative disease in a subject, the methods comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition, wherein the pharmaceutical comprises a pharmaceutically acceptable form of Compound 1, and wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form disclosed herein. In some embodiments, the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, Huntington's disease, schizophrenia or autism.

[0067] The present invention further provides a composition for treating cancer in a subject, the composition comprising a form of Compound 1. In some embodiments, the composition comprises the succinate form of Compound 1. In some embodiments, the composition comprises the gentisate form of Compound 1. In some embodiments, the composition comprises the gentisate form of Compound 1, wherein the form is crystalline. In some embodiments, the composition comprises the gentisate form of Compound 1, wherein the form is a crystalline solid that exhibits a peak at 9.3 ± 0.2° 2-θ in an X-ray powder diffraction (XRPD) pattern. In still further embodiments, the crystalline solid exhibits a further peak at 7.1 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In still further embodiments, the crystalline solid exhibits a further peak at 20.1 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In still further embodiments, the crystalline solid exhibits a further peak at 19.0 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In still further embodiments, the crystalline solid exhibits a further peak at 26.2 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In still further embodiments, the crystalline solid exhibits further peaks at 4.8 ± 0.2° 2-θ, 10.5 ± 0.2° 2-θ, and 14.8 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern. In some embodiments, the cancer is lung cancer, melanoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, prostate cancer, kidney cancer, multiple myeloma, or lymphoma. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is melanoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is gallbladder cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the prostate cancer is castration-resistant prostate, metastatic prostate cancer, metastatic castration-resistant prostate cancer, or castration-sensitive prostate cancer. In some embodiments, the cancer is kidney cancer. In some embodiments, the cancer is multiple myeloma. In some embodiments, the multiple myeloma is light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma, or immunoglobulin E (IgE) myeloma. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer expresses CD73. In some embodiments, CD73 is upregulated in the cancer to be treated.In some embodiments, the cancer in which CD73 is expressed or upregulated is lung cancer, melanoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, prostate cancer, renal cancer, multiple myeloma, or lymphoma. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is melanoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is gallbladder cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the prostate cancer is castration-resistant prostate, metastatic prostate cancer, metastatic castration-resistant prostate cancer, or castration-sensitive prostate cancer. In some embodiments, the cancer is renal cancer. In some embodiments, the cancer is multiple myeloma. In some embodiments, the multiple myeloma is light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma, or immunoglobulin E (IgE) myeloma. In some embodiments, the cancer is lymphoma.

[0068] The present disclosure also provides a pharmaceutical composition comprising a pharmaceutically acceptable form of a compound 1 in an amount and one or more pharmaceutically acceptable excipients, wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form. In some embodiments, the gentisate form comprises gentisate. In some embodiments, the gentisate form includes a gentisate co-crystal. In some embodiments, the gentisate form comprises a salt and a co-crystal. In some embodiments, the succinate form comprises succinate. In some embodiments, the succinate form comprises a succinate co-crystal. In some embodiments, the succinate form comprises a salt and a co-crystal. The present disclosure further provides such a pharmaceutical composition, wherein the one or more pharmaceutically acceptable excipients include one or more diluents, binders, disintegrants, lubricants, anti-adhesives, glidants, colorants, flavorants, sweeteners, coating agents, plasticizers, wetting agents, buffers, or adsorbents.

[0069] One or more diluents that can be used include lactose, mannitol, xylitol, microcrystalline cellulose, calcium hydrogen phosphate, and starch. In some embodiments, the one or more diluents comprise from about 1% to about 80%, or from about 10% to about 80%, or from about 10% to about 70%, or from about 15% to about 80%, or from about 20% to about 80%, or from about 15% to about 75%, or from about 20% to about 75%, or from about 25% to about 75%, or from about 50% to about 80%, or from about 50% to about 75%, or from about 60% to about 80%, or from about 60% to 75% of the total weight of the pharmaceutical composition. In some embodiments, the diluent is lactose. In some embodiments, the diluent is mannitol. In some embodiments, the diluent is xylitol. In some embodiments, the diluent is microcrystalline cellulose. In some embodiments, the diluent is calcium hydrogen phosphate. In some embodiments, the diluent is starch.

[0070] The present disclosure also provides such pharmaceutical compositions, wherein the one or more pharmaceutically acceptable excipients comprise one or more binders, and wherein the one or more binders comprise from about 1% to about 80%, or from about 10% to about 80%, or from about 10% to about 70%, or from about 15% to about 80%, or from about 20% to about 80%, or from about 15% to about 75%, or from about 20% to about 75%, or from about 25% to about 75%, or from about 50% to about 80%, or from about 50% to about 75%, or from about 60% to about 80%, or from about 60% to 75% of the total weight of the pharmaceutical composition. In some embodiments, the one or more binders are selected from methylcellulose, microcrystalline cellulose, starch, and gums (such as guar gum and tragacanth gum) or mixtures thereof.

[0071] The present invention also provides such pharmaceutical compositions, wherein one or more pharmaceutically acceptable excipients comprise one or more disintegrants, and wherein the one or more disintegrants account for about 0.1% to about 10%, or about 0.1% to about 5%, or about 0.1% to about 4%, or about 0.1% to about 3%, or about 0.1% to about 2%, or about 0.1% to about 1%, or about 0.1% to about 0.75%, or about 0.2% to about 1%, or about 0.3% to about 1%, or about 0.4% to about 1%, or about 0.2% to about 0.8%, or about 0.3% to about 0.75%, or about 0.3% to about 0.7%, or about 0.3% to about 0.6% by weight of the total weight of the pharmaceutical composition. In some embodiments, the one or more disintegrants are selected from starch, sodium starch glycolate, sodium alginate, sodium carboxymethylcellulose, methylcellulose, croscarmellose sodium, and crospovidone or mixtures thereof. In some embodiments, the disintegrant is starch. In some embodiments, the disintegrant is sodium starch glycolate. In some embodiments, the disintegrant is sodium alginate. In some embodiments, the disintegrant is sodium carboxymethylcellulose. In some embodiments, the disintegrant is methylcellulose. In some embodiments, the disintegrant is croscarmellose sodium. In some embodiments, the disintegrant is crospovidone.

[0072] The present invention also provides such pharmaceutical compositions, wherein one or more pharmaceutically acceptable excipients comprise one or more lubricants, and wherein the one or more lubricants account for about 0.1% to about 10%, or about 0.1% to about 5%, or about 0.1% to about 4%, or about 0.1% to about 3%, or about 0.1% to about 2%, or about 0.1% to about 1%, or about 0.1% to about 0.75%, or about 0.2% to about 1%, or about 0.3% to about 1%, or about 0.4% to about 1%, or about 0.2% to about 0.8%, or about 0.3% to about 0.75%, or about 0.3% to about 0.7%, or about 0.3% to about 0.6% by weight of the total weight of the pharmaceutical composition. In further embodiments, the one or more lubricants are selected from magnesium stearate, calcium stearate, sodium stearyl fumarate, and stearic acid or mixtures thereof. In some embodiments, the lubricant is magnesium stearate. In some embodiments, the lubricant is calcium stearate. In some embodiments, the lubricant is sodium stearyl fumarate. In some embodiments, the lubricant is stearic acid.

[0073] In some embodiments, the pharmaceutical compositions disclosed herein may comprise additional excipients including, but not limited to, buffers, glidants, preservatives, and colorants. Additional excipients such as fillers, tonicity agents, and chelating agents are also within the scope of the embodiments.

[0074] Non-limiting examples of buffering agents include, but are not limited to, sodium bicarbonate, potassium bicarbonate, magnesium hydroxide, magnesium lactate, magnesium gluconate, aluminum hydroxide, aluminum hydroxide / sodium bicarbonate coprecipitate, mixtures of amino acids and buffering agents, mixtures of glycine aluminum and buffering agents, acid salts of amino acids and buffering agents, and basic salts of amino acids and buffering agents. Additional buffering agents include sodium citrate, sodium tartrate, sodium acetate, sodium carbonate, sodium polyphosphate, potassium polyphosphate, sodium pyrophosphate, potassium pyrophosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, trisodium phosphate, tripotassium phosphate, sodium acetate, potassium metaphosphate, magnesium oxide, magnesium hydroxide, magnesium carbonate, magnesium silicate, calcium acetate, calcium glycerophosphate, calcium chloride, calcium hydroxide, calcium lactate, calcium carbonate, calcium bicarbonate, and other calcium salts.

[0075] In some embodiments, the pharmaceutical compositions disclosed herein may contain glidants. Suitable glidants include, but are not limited to, tricalcium phosphate, calcium silicate, cellulose, colloidal silicon dioxide, magnesium silicate, magnesium trisilicate, silicon dioxide, starch, talc, and the like. In some embodiments, the glidant is tricalcium phosphate. In some embodiments, the glidant is calcium silicate. In some embodiments, the glidant is cellulose. In some embodiments, the glidant is colloidal silicon dioxide. In some embodiments, the glidant is magnesium silicate. In some embodiments, the glidant is magnesium trisilicate. In some embodiments, the glidant is silicon dioxide. In some embodiments, the glidant is starch. In some embodiments, the glidant is talc.

[0076] In some embodiments, the pharmaceutical compositions disclosed herein may contain preservatives. Preservatives include antimicrobial agents, antioxidants, and agents that enhance sterility. Exemplary preservatives include ascorbic acid, ascorbyl palmitate, BHA, BHT, citric acid, isoascorbic acid, fumaric acid, malic acid, propyl gallate, sodium ascorbate, sodium bisulfate, sodium metabisulfite, sodium sulfite, parabens (methyl-, ethyl-, butyl-), benzoic acid, potassium sorbate, vanillin, and the like.

[0077] In some embodiments, the pharmaceutical compositions disclosed herein may contain colorants for identification and / or aesthetic purposes of the resulting liquid form. Suitable colorants illustratively include FD&C Red No. 3, FD&C Red No. 20, FD&C Red No. 40, FD&C Yellow No. 6, FD&C Blue No. 2, D&C Green No. 5, D&C Orange No. 5, caramel, iron oxide, and mixtures thereof.

[0078] Additional excipients are contemplated in the pharmaceutical compositions disclosed herein. These additional excipients are selected based on function and compatibility with the pharmaceutical compositions described herein and can be found, for example, in Remington: The Science and Practice of Pharmacy, 19th Edition (Easton, PA: Mack Publishing Company, 1995), which is incorporated herein by reference in its entirety; Hoover, John E., Remington’s Pharmaceutical Sciences, (Easton, PA: Mack Publishing Co 1975); Liberman, H.A. and Lachman, L., eds., Pharmaceutical Dosage Forms (New York, NY: Marcel Decker 1980); and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition (Lippincott Williams & Wilkins 1999).

[0079] The pharmaceutical compositions disclosed herein can be in a form suitable for oral administration to a subject in need thereof. Suitable oral dosage forms include, for example, tablets, pills, cachets or capsules of hard or soft gelatin, methylcellulose or another suitable material that is readily soluble in the digestive tract. Oral administration of solid dosage forms can be presented, for example, in discrete units such as hard or soft capsules, pills, lozenges, troches or tablets, each containing a predetermined amount of the compound 1 as disclosed herein and one or more pharmaceutically acceptable excipients. In another embodiment, oral administration can be in the form of a powder or granules. In another embodiment, the oral dosage form is a sublingual dosage form, such as, for example, a lozenge. Capsules or tablets can contain controlled release formulations. In the case of capsules, tablets and pills, the dosage form can also contain buffering agents or can be prepared to have an enteric coating. In another embodiment, oral administration can be in a liquid dosage form. Liquid dosage forms for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing inert diluents commonly used in the art (e.g., water). Such compositions can also contain adjuvants such as wetting agents, emulsifying agents, suspending agents, flavoring agents (e.g., sweeteners) and / or fragrances.

[0080] The present invention also provides tablets comprising a pharmaceutically acceptable form of Compound 1, microcrystalline cellulose, colloidal silicon dioxide, sodium stearyl fumarate, crospovidone, and magnesium stearate. In some embodiments, the pharmaceutically acceptable form of Compound 1 is selected from the gentisate form and the succinate form. In some embodiments, the pharmaceutically acceptable form of Compound 1 is the gentisate form. In some embodiments, the gentisate form comprises a salt. In some embodiments, the gentisate form comprises a cocrystal. In some embodiments, the gentisate form comprises a salt and a cocrystal. In some embodiments, the pharmaceutically acceptable form of Compound 1 is the succinate form. In some embodiments, the succinate form comprises a salt. In some embodiments, the succinate form comprises a cocrystal. In some embodiments, the succinate form comprises a salt and a cocrystal. In a further embodiment, the tablets disclosed herein are as shown in Table 1.

[0081] Table 1

[0082] Material Quantity / tablet (mg) Weight percentage of tablet Compound 1 gentisate form 261.438 38.45％ Microcrystalline cellulose PH 377.162 55.46％ Colloidal silica 7.000 1.03％ Sodium stearyl fumarate 7.000 1.03％ Crospovidone 20.400 3.00％ Magnesium stearate 7.000 1.03％ Total 680 100％

[0083] In another embodiment, the pharmaceutical compositions disclosed herein may include parenteral dosage forms. "Parenteral administration" includes, for example, subcutaneous injection, intravenous injection, intraperitoneal injection, intramuscular injection, intracardiac injection, and infusion. Injectable preparations (e.g., sterile injectable aqueous or oily suspensions) may be formulated according to known techniques using suitable dispersing, wetting, and / or suspending agents.

[0084] In another embodiment, the pharmaceutical compositions disclosed herein may include topical dosage forms. "Topical administration" includes, for example, transdermal administration (such as via a transdermal patch or iontophoresis device), intraocular administration, or intranasal or inhaled administration. Compositions for topical administration also include, for example, topical gels, sprays, ointments, and creams. Topical formulations may include compounds that enhance the absorption or penetration of the active ingredient through the skin or other affected area. When the compositions disclosed herein are administered via a transdermal device, the administration will be achieved using patches of the reservoir and porous membrane type or the solid matrix variety. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, powders, dressings, foams, membranes, skin patches, sheets, implants, sponges, fibers, bandages, and microemulsions. Liposomes may also be used. Typical carriers include alcohols, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. Penetration enhancers may be incorporated; see, e.g., J. Pharm. Sci., 88(10), 955-958, by Finnin and Morgan (October 1999).

[0085] For intranasal administration or administration by inhalation, the pharmaceutical compositions disclosed herein are conveniently delivered from a pump spray container as a solution or suspension by patient pressing or pumping, or presented as an aerosol spray from a pressurized container or nebulizer using a suitable propellant. Formulations suitable for intranasal administration are generally administered in the form of a dry powder from a dry powder inhaler (alone; as a mixture, e.g., in a dry blend with lactose; or as mixed component particles, e.g., mixed with a phospholipid such as phosphatidylcholine), or as an aerosol spray from a pressurized container, pump, nebulizer, atomizer (preferably an electrohydrodynamic atomizer to produce a fine mist) or nebulizer, with or without a suitable propellant such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may contain a bioadhesive, such as chitosan or cyclodextrin.

[0086] Other carrier materials and modes of administration known in the pharmaceutical art may also be used. The pharmaceutical compositions disclosed herein can be prepared by any well-known pharmaceutical technique, such as effective formulation and administration procedures. The above considerations regarding effective formulation and administration procedures are well-known in the art and are described in standard textbooks. The formulation of drugs is discussed, for example, in Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1975; Liberman et al., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Kibbe et al., eds., Handbook of Pharmaceutical Excipients (3rd ed.), American Pharmaceutical Association, Washington, 1999.

[0087] The dosage of a composition comprising a form of Compound 1 as described herein may vary depending on the condition of the patient (e.g., a human), i.e., the stage of the disease, general health status, age, and other factors. The pharmaceutical composition is administered in a manner suitable for the disease to be treated (or prevented). The appropriate dosage, as well as the suitable duration and frequency of administration, will be determined by factors such as the condition of the patient, the type and severity of the patient's disease, the specific form of the active ingredient, and the method of administration. Generally, the appropriate dosage and treatment regimen provide one or more compositions in an amount sufficient to provide a therapeutic and / or prophylactic benefit (e.g., improvement in clinical outcomes such as more frequent complete or partial remission, or longer disease-free and / or overall survival, or reduction in symptom severity). The optimal dosage is typically determined using experimental models and / or clinical trials. The optimal dosage depends on the body mass, weight, or blood volume of the patient. Oral dosages are generally in the range of about 1.0 mg to about 1000 mg, one to four or more times per day.

[0088] Compound 1 can be prepared by methods known to those of ordinary skill in the art, including but not limited to the methods described in Example 1.

[0089] Abbreviations used in the examples include: rt (room temperature), min (minute), h (hour), MeCN (acetonitrile), DMF (N,N-dimethylformamide), THF (tetrahydrofuran), MeOH (methanol), sat (saturated), and TsOH (tosylic acid).

[0090] Example

[0091] Example 1. Preparation of ((S)-1-((2H-Tetrazol-5-yl)methoxy)-2-(((2R,3S,4R,5R)-5-(6-Chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-hydroxypropan-2-yl)phosphonic Acid

[0092]

[0093] Step A. (2R,3R,4R,5R)-2-(Acetoxymethyl)-5-(4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)tetrahydrofuran-3,4-diyl diacetate (1a)

[0094]

[0095] At 90 °C, 1,2,3,5 - tetra - O - acetyl - β - D - ribofuranose (5.73 g, 17.99 mmol) was heated for 10 min, and 4,6 - dichloro - 1H - pyrazolo[3,4 - d]pyrimidine (1.5 g, 17.99 mmol) and SnCl4 (60 mg) were added successively. After heating the mixture under reduced pressure at 130 °C for 15 min, it was cooled to room temperature, diluted with water, and extracted with DCM. The combined organic phases were washed with water and brine, dried, and concentrated. The residue was purified by column chromatography (petroleum ether / ethyl acetate, from 10:1 to 5:1) to give the title compound (1a) as a yellow solid (2.4 g, 68%).

[0096] Step B. (2R,3R,4R,5R) - diacetate 2 - (acetoxymethyl) - 5 - (6 - chloro - 4 - (cyclopentylamino) - 1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)tetrahydrofuran - 3,4 - diyl ester (1b)

[0097]

[0098] 1a (5.2 g, 11.63 mmol) was added to a dried flask, followed by ethanol (53.24 mL). Triethylamine (2.43 mL, 17.44 mmol) was added to this solution, followed by cyclopentylamine (1.38 mL, 13.95 mmol). After stirring and heating the mixture at 50 °C for 15 min, it was cooled to room temperature, concentrated, and purified by column chromatography (20 to 45% ethyl acetate / hexane, gradient elution) to afford the title compound (1b) as a white solid (5.02 g, 87%). m / z (ESI, positive ion) = 496.1 [M + H] + 。

[0099] Step C. (2R,3R,4R,5R) - diacetate 2 - (acetoxymethyl) - 5 - (4 - ((tert - butoxycarbonyl)(cyclopentyl)amino) - 6 - chloro - 1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)tetrahydrofuran - 3,4 - diyl ester (1c)

[0100]

[0101] Triethylamine (5.14 g, 50.9 mmol) was added to a solution of 1b (12.6 g, 25.4 mmol) in MeCN (120 mL), followed by di-tert-butyl dicarbonate (44.35 g, 203.6 mmol) and 4-dimethylaminopyridine (0.31 g, 2.54 mmol). After stirring the mixture overnight, it was concentrated and partitioned between EtOAc (50 mL) and saturated NaHCO3. The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether / ethyl acetate: 8:1) to afford the title compound (1c) as a yellow solid (10.56 g, 70% yield). m / z (ESI, positive ion) = 596.72 [M+H] + 。

[0102] Step D. (6-Chloro-1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamic acid tert-butyl ester (1d)

[0103]

[0104] 1c (10.56 g, 17.78 mmol) was added to a dried flask, followed by a methanol solution of ammonia (5.0 M, 140 mL). The mixture was stirred overnight and then concentrated. The crude oil was purified by column chromatography to afford the title compound (1d) as a yellow solid (7.39 g, 89% yield). m / z (ESI, positive ion) = 470.3 [M+H] + 。

[0105] Step E. (6-Chloro-1-((3aR,4R,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamic acid tert-butyl ester (1e)

[0106]

[0107] TsOH was added to a solution of 1d (7.39 g, 15.75 mmol) and 2,2-dimethoxypropane (4.92 g, 47.27 mmol) in DMF (75 mL) .H2O (0.6 g, 3.15 mmol). After stirring the mixture at 70 °C for 1 h, it was cooled and quenched with saturated NaHCO3 (100 mL). The mixture was extracted with EtOAc (50 mL), and the combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated. The crude oil was purified by column chromatography (petroleum ether / ethyl acetate: 8:1) to afford the title compound (1e) as a yellow solid (5.5 g, 68% yield). m / z (ESI, positive ion) = 510.4 [M+H] + .

[0108] Step F. Ethyl 2-(((3aR,4R,6R,6aR)-6-(4-((tert-butoxycarbonyl)(cyclopentyl)amino)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-(diethoxyphosphoryl)acetate (1f)

[0109]

[0110] Under N2, Rh2(OAc)4 (0.96 g, 2.17 mmol) was added to a solution of ethyl 2-diazo-2-(diethoxyphosphoryl)acetate (13.5 g, 54.13 mmol) and 1e (5.5 g, 10.83 mmol) in toluene (80 mL). After stirring the mixture at 95 °C overnight, it was concentrated and purified by column chromatography (petroleum ether / ethyl acetate: 5:1) to afford the title compound (1f) as a yellow oil (6 g, 76% yield). m / z (ESI, positive ion) = 732.2 [M+H] + .

[0111] Step G. Ethyl 2-[[(3aR,4R,6R,6aR)-4-[4-[tert-butoxycarbonyl(cyclopentyl)amino]-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl]-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methoxy]-2-diethoxyphosphoryl-3-(2-trimethylsilylethoxy)propionate (1g)

[0112]

[0113] At -15 °C, sodium bis(trimethylsilyl)amide (1.0 M in THF, 2.13 mL, 2.13 mmol) was added dropwise to a solution of compound 1f (1.2 g, 1.64 mmol) in THF (33 mL). After stirring for 25 min at -15 °C, tetrabutylammonium iodide (303 mg, 0.820 mmol) was added, and immediately followed by dropwise addition of 2-(chloromethoxy)ethyl](trimethyl)silane (0.863 mL, 4.92 mmol) to the solution. The mixture was stirred for 1 h at the same temperature and then quenched with saturated aqueous NH4Cl. The solution was diluted with EtOAc and water and extracted with EtOAc. The combined organic layers were washed (brine), dried (Na2SO4), and concentrated under reduced pressure. Purification of the residue by silica gel column chromatography (5 - 30% acetone / hexane, gradient elution) afforded the title compound as a pale yellow oil (1 g) (1.03 g, 73%). m / z (ESI, positive ion) = 862.3 [M+H] + .

[0114] Step H. tert-Butyl N-[1-[(3aR,4R,6R,6aR)-6-[[1-diethoxyphosphoryl-1-(hydroxymethyl)-2-(2-trimethylsilylethoxy)ethoxy]methyl]-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-4-yl]-6-chloropyrazolo[3,4-d]pyrimidin-4-yl]-N-cyclopentylcarbamate (1h)

[0115]

[0116] At 0 °C, calcium dichloride (596 mg, 5.37 mmol) was added to a stirred solution of ethyl 2-[[(3aR,4R,6R,6aR)-4-[4-[tert-butoxycarbonyl(cyclopentyl)amino]-6-chloro-pyrazolo[3,4-d]pyrimidin-1-yl]-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methoxy]-2-diethoxyphosphoryl-3-(2-trimethylsilylethoxy)propionate (1 g) (1.03 g, 1.19 mmol) in EtOH (18 mL), and then sodium borohydride (203 mg, 5.37 mmol) was added in one portion. The mixture was warmed to room temperature and stirred for 3 h. Then the mixture was cooled back to 0 °C and the mixture was quenched with 1 N aqueous HCl, diluted with EtOAc and water. The solution was extracted (EtOAc), and the combined organic layers were washed (brine), dried (Na2SO4), and concentrated under reduced pressure. Purification of the residue by silica gel column chromatography (1-5% MeOH / DCM, gradient elution) afforded the title compound (1 h) as a white foamy solid (755 mg, 77%). m / z (ESI, positive ion) = 820.3 [M+H]+.

[0117] Step I. (6-Chloro-1-((3aR,4R,6R,6aR)-6-(((2-(diethoxyphosphoryl)-1-(2-(trimethylsilyl)ethoxy)-3-((2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)methoxy)propan-2-yl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamic acid tert-butyl ester (1 i)

[0118]

[0119] At 0 °C, NaH (60% mineral oil, 77.0 mg, 1.93 mmol) was added in one portion to a solution of tert-butyl N-[1-[(3aR,4R,6R,6aR)-6-[[1-diethoxyphosphoryl-1-(hydroxymethyl)-2-(2-trimethylsilylethoxy)ethoxy]methyl]-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-4-yl]-6-chloro-pyrazolo[3,4-d]pyrimidin-4-yl]-N-cyclopentyl-carbamate (1h) (632 mg, 0.770 mmol) and 5-(bromomethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazole (1.13 g, 3.85 mmol) in DMF (5.0 mL). After stirring the mixture at 0 °C for 30 min, the mixture was quenched with saturated aqueous NH4Cl, diluted with EtOAc and water. The solution was extracted (EtOAc), and the combined organic layers were washed (brine), dried (Na2SO4), and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (5 - 30% acetone / hexane, gradient elution) to afford the title compound (1i) as a pale yellow gum (697 mg, 88%).

[0120] Step J. (6-Chloro-1-((3aR,4R,6R,6aR)-6-((((R)-2-(diethoxyphosphoryl)-1-(2-(trimethylsilyl)ethoxy)-3-((2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)methoxy)propan-2-yl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamate (1j)

[0121]

[0122] The diastereomers from Step I were separated by chiral chromatography (CHIRALPAK, AD-H, 21×250 mm, 5 μm, 5% IPA / hexane, isocratic elution, flow rate of 20 mL / min), and the second eluted isomer was identified as the title compound (1j) and collected.

[0123] Step K. Diethyl ((S)-1-((2H-tetrazol-5-yl)methoxy)-2-(((3aR,4R,6R,6aR)-6-(6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-3-hydroxypropan-2-yl)phosphonate (1k)

[0124]

[0125] At 0 °C, boron trifluoride etherate (0.233 mL, 1.89 mmol) was added dropwise to a solution of tert-butyl (6-chloro-1-((3aR,4R,6R,6aR)-6-((((R)-2-(diethoxyphosphoryl)-1-(2-(trimethylsilyl)ethoxy)-3-((2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)methoxy)propan-2-yl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamate (1j) (325 mg, 0.315 mmol) in DCM (16 mL). The reaction was warmed to room temperature. After stirring at room temperature for 3.5 h, the reaction was quenched with triethylamine (3.6 mL) and the resulting mixture was stirred at room temperature for 10 min. Saturated aqueous NaHCO3 (7.2 mL) was added to the mixture and the solution was diluted with DCM and water. The solution was extracted (DCM) and the combined organic layers were washed (brine), dried (Na2SO4), and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (0 - 20% MeOH / DCM, gradient elution) to afford the title compound (1k) as an off-white foamy solid (189 mg, 86%). m / z (ESI, positive ion) = 702.3 [M+H]+.

[0126] Step L. ((S)-1-((2H-tetrazol-5-yl)methoxy)-2-(((2R,3S,4R,5R)-5-(6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-hydroxypropan-2-yl)phosphonic acid (1)

[0127]

[0128] At room temperature, under an argon atmosphere, triethylamine (0.751 mL, 5.38 mmol) was added to a solution of ((S)-1-((2H-tetrazol-5-yl)methoxy)-2-(((3aR,4R,6R,6aR)-6-(6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-3-hydroxypropan-2-yl) diethylphosphonate (1k) (189 mg, 0.269 mmol) in MeCN (13.5 mL), followed by the addition of bromotrimethylsilane (0.528 mL, 4.04 mmol). After stirring the solution at room temperature for 4 h, it was concentrated under reduced pressure. The residue was dissolved in TFA / H2O (1 / 3, 10 mL) and stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase HPLC (15 - 40% ACN / H2O, 0.1% TFA, gradient elution) to afford the title compound (1) as an off-white solid (TFA salt, 107 mg, 55%). 1H NMR (400 MHz, methanol-d4) δ 8.08 (d, J = 0.8 Hz, 1H), 6.25 - 6.20 (m, 1H), 4.96 (s, 2H), 4.72 - 4.69 (m, 1H), 4.57 - 4.47 (m, 2H), 4.19 - 4.16 (m, 1H), 4.08 (dd, J = 10.4, 4.0 Hz, 1H), 4.01 - 3.92 (m, 4H), 3.84 (dd, J = 12.4, 7.6 Hz, 1H), 2.13 - 2.06 (m, 2H), 1.84 - 1.57 (m, 6H); m / z (ESI, positive ion) = 606.1 [M+H] + .

[0129] Alternatively, Example 1 ((S)-1-((2H-tetrazol-5-yl)methoxy)-2-(((2R,3S,4R,5R)-5-(6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-hydroxypropan-2-yl) phosphonic acid was prepared by the following steps M to O.

[0130] Step M. tert-Butyl (6-chloro-1-((3aR,4R,6R,6aR)-6-((((R)-2-(diethoxyphosphoryl)-1-hydroxy-3-(2-(trimethylsilyl)ethoxy)propan-2-yl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamate (1m)

[0131]

[0132] The diastereomers from step H (1 h) were separated by chiral chromatography (CHIRALPAK, AD-H, 21×250 mm, 5 μm, 5% IPA / hexane, isocratic elution, flow rate of 20 mL / min), and the second eluted isomer was identified as the title compound (1 m) and collected.

[0133] Step N. tert-Butyl (6-chloro-1-((3aR,4R,6R,6aR)-6-((((R)-2-(diethoxyphosphoryl)-1-(2-(trimethylsilyl)ethoxy)-3-((2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)methoxy)propan-2-yl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamate (1 n)

[0134]

[0135] At 0 °C, NaH (60% mineral oil, 252 mg, 6.31 mmol) was added in one portion to a solution of tert-butyl (6-chloro-1-((3aR,4R,6R,6aR)-6-((((R)-2-(diethoxyphosphoryl)-1-hydroxy-3-(2-(trimethylsilyl)ethoxy)propan-2-yl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamate (1 m) (2.07 g, 2.52 mmol) and 5-(bromomethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazole (2.96 g, 10.1 mmol) in DMF (12.5 mL). After stirring the mixture at 0 °C for 30 min, the mixture was quenched with saturated aqueous NH4Cl, diluted with EtOAc and water. The solution was extracted (EtOAc), and the combined organic layers were washed (brine), dried (Na2SO4), and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (5 - 30% acetone / hexane, gradient elution) to afford the title compound (1 n) as a pale yellow gum (2.2 g, 84%).

[0136] Step O. ((S)-1-((2H-Tetrazol-5-yl)methoxy)-2-(((2R,3S,4R,5R)-5-(6-Chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-hydroxypropan-2-yl)phosphonic acid (1)

[0137]

[0138] At room temperature, under an argon atmosphere, triethylamine (16.2 mL, 116 mmol) was added to a solution of tert-butyl (6-chloro-1-((3aR,4R,6R,6aR)-6-((((R)-2-(diethoxyphosphoryl)-1-(2-(trimethylsilyl)ethoxy)-3-((2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)methoxy)propan-2-yl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)(cyclopentyl)carbamate (1n) (8.00 g, 7.75 mmol) in MeCN (300 mL), followed by the addition of bromotrimethylsilane (10.1 mL, 77.5 mmol). After stirring the solution for 15 h, it was concentrated under reduced pressure and co-distilled with toluene (2 times). The residue was partitioned between EtOAc and water. The organic layer was collected, and the aqueous layer was extracted with EtOAc twice. The combined organic layers were washed with water again and concentrated under reduced pressure. The crude solid was dissolved in TFA / water (1 / 1, 280 mL) and stirred at room temperature for 24 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase HPLC (15 - 35% ACN / H2O, 0.1% TFA, gradient elution) to afford the title compound (1) as a white solid (TFA salt, 3.1 g, 56%). 1H NMR (400 MHz, methanol-d4) δ 8.08 (d, J = 0.8 Hz, 1H), 6.25 - 6.20 (m, 1H), 4.96 (s, 2H), 4.72 - 4.69 (m, 1H), 4.57 - 4.47 (m, 2H), 4.19 - 4.16 (m, 1H), 4.08 (dd, J = 10.4, 4.0 Hz, 1H), 4.01 - 3.92 (m, 4H), 3.84 (dd, J = 12.4, 7.6 Hz, 1H), 2.13 - 2.06 (m, 2H), 1.84 - 1.57 (m, 6H); m / z (ESI, positive ion) = 606.1 [M+H] + 。

[0139] Example 2: Preparation of the succinate form of Compound 1

[0140] Prepare a slurry containing a certain amount of Compound 1 (20 mg, free base) and an equimolar amount of succinic acid in tetrahydrofuran / water (about 0.2 mL, 9:1, v / v), and stir the resulting slurry at room temperature for 3 days and then at 5 °C for another 3 days to obtain a clear solution. Then add a certain amount of acetonitrile (about 5 mL) to obtain a clear solution, which is transferred to an open container. Evaporate the solvent to provide a solid precipitate, and collect the precipitate to obtain Compound 1 in the form of succinate salt.

[0141] Example 3: Preparation of Form 1 of the gentisate salt form of Compound 1

[0142] Place a certain amount of Compound 1 (20 mg, free base) and an equimolar amount of gentisic acid in THF / H2O (9:1, v / v) into a vial, and stir the resulting mixture at room temperature for three days and then at 5 °C for another 3 days to obtain a clear solution. Then add a certain amount of acetonitrile (about 5 mL) to obtain a clear solution, which is transferred to an open container. Evaporate the solvent to provide a solid precipitate, and collect the precipitate to obtain Form 1 of the gentisate salt form of Compound 1.

[0143] Example 4: Preparation of Form 2 of the gentisate salt form of Compound 1

[0144] Add a certain amount of Compound 1 (50.2 mg, free base) and gentisic acid (12.5 mg) to a 5-mL glass vial, add 1 mL of a tetrahydrofuran / water (9:1, v / v) mixture thereto to obtain a slurry, and stir it at room temperature for 1 day until a clear solution is obtained. Add a certain amount of acetonitrile (3 mL) to the solution to obtain a clear solution. Then evaporate the solvent at room temperature to provide a solid precipitate, and collect the precipitate to obtain Form 2 of the gentisate salt form of Compound 1.

[0145] Example 5: Preparation of Form 2 of the gentisate salt form of Compound 1

[0146] To a solution of the trifluoroacetic acid form of Compound 1 from Example 1 in a water / n-propanol (2:98, 3 volumes) mixture, gentisic acid (3 molar equivalents) was added and the resulting mixture was stirred at 25 °C. A seed crystal of Form 2 of the gentisate form of Compound 1 (prepared according to a method similar to Example 4) (1 mol %) was added to the resulting solution and the resulting mixture was stirred for >1 h. Then, heptane (2 volumes) was added over >1 h and the resulting mixture was stirred for >12 h. Additional heptane (19 volumes) was added to the mixture over >10 h and the mixture was stirred for >8 h. The mixture was filtered, the solid was washed three times with a water / n-propanol / heptane mixture (1:37:100, 2 volumes), deliquored, and dried at 40 °C under vacuum, and the humidity inside the dryer was controlled at a relative humidity of 25% to 45% to obtain Form 2 of the gentisate form of Compound 1.

[0147] Example 6: X-ray powder diffraction (XRPD) analysis of the polymorphic forms of the gentisate form of Compound 1

[0148] XRPD analysis of the polymorphic forms of the gentisate form of Compound 1 was performed using a Panalytical Empyrean and an X’pert3 X-ray powder diffractometer. The sample was spread in the middle of a zero-background Si holder. The parameters used for the analysis are shown in Table 2.

[0149] Table 2

[0150]

[0151] Form 1 of the gentisate form of Compound 1 was analyzed by XRPD as described above and exhibited the peaks shown in Table 3. The error associated with each °2θ position was determined to be ±0.2°θ.

[0152] Table 3

[0153]

[0154] Form 2 of the gentisate form of Compound 1 was analyzed by XRPD as described above and exhibited the peaks shown in Table 4. The error associated with each °2θ position was determined to be ±0.2°θ.

[0155] Table 4

[0156]

[0157] Example 7: Thermogravimetric analysis and differential scanning calorimetry analysis of the gentisate form of Compound 1

[0158] Thermogravimetric analysis (TGA) data were collected using a TA Q5000 and Discovery TGA 5500 TGA from TA Instruments. Differential scanning calorimetry (DSC) analysis was performed using a TA Q2000 DSC from TA Instruments with the parameters shown in Table 5.

[0159] Table 5

[0160] Parameter TGA DSC Method Ramp Ramp Sample pan Aluminum, open Aluminum, open Temperature range Room temperature to about 350 °C 25 °C to about 200 °C Heating rate 10 °C / min 10 °C / min Purge gas Nitrogen Nitrogen

[0161] Example 8: Solubility of the Polymorphic Forms of the Gentisate Salt Form of Compound 1

[0162] As described below, the solubility of the polymorphic forms of the gentisate salt form of Compound 1 was measured after 4 hours and at 37 °C in water, simulated gastric fluid (SGF), fasted-state simulated intestinal fluid (FaSSIF), and fed-state simulated intestinal fluid (FeSSIF).

[0163] The SGF medium was prepared as follows: 49.5 mg of NaCl and 25.4 mg of Triton X-100 were weighed into a 100 mL volumetric flask. A certain volume of purified water was added to the flask and the resulting mixture was sonicated until all solids were dissolved. Then approximately 1.632 mL of HCl solution (1 M) was added and sufficient purified water was added to the target volume and the pH was adjusted to 1.8. The solution was then diluted to a certain volume with purified water, mixed well, and the pH was measured to be 1.83.

[0164] The FaSSIF buffer was prepared as follows: 340.8 mg of NaH2PO4, 43.0 mg of NaOH, and 619.6 mg of NaCl were weighed into a 100-mL volumetric flask. A certain volume of purified water was added to the flask and the resulting mixture was sonicated until the solids were dissolved. A second volume of purified water was added to the flask to adjust the pH to 6.5. The solution was diluted with another volume of purified water, mixed, and the pH was measured to be 6.54. The FaSSIF medium was prepared as follows: 110.4 mg of SIF powder was weighed into a 50-mL volumetric flask and a certain volume of FaSSIF dissolution buffer was added thereto. The resulting mixture was sonicated until the SIF powder was dissolved. The mixture was then diluted to a certain volume with FaSSIF dissolution buffer and mixed well. Before use, the FaSSIF solution was equilibrated at room temperature for 2 hours.

[0165] The FeSSIF dissolution buffer was prepared as follows: 0.82 mL of glacial acetic acid, 404.9 mg of NaOH, and 1188.2 mg of NaCl were weighed into a 100-mL volumetric flask. A certain volume of purified water was added to the flask and the mixture was sonicated until the solids dissolved. A second volume of purified water was added to the target volume to adjust the pH to 5.0. The solution was diluted with a certain volume of purified water, mixed well, and the pH was measured to be 4.96. The FeSSIF medium was prepared as follows: 559.6 mg of SIF powder was weighed into a 50-mL volumetric flask. A certain volume of FeSSIF dissolution buffer was added to the flask, and the resulting mixture was sonicated to dissolve the SIF powder. A second volume of FeSSIF dissolution buffer was added and the resulting mixture was mixed well. Before use, the FeSSIF solution was equilibrated for 2 hours to room temperature.

[0166] Solubility measurements were performed using an Agilent 1260 high-performance liquid chromatography (HPLC) instrument equipped with a DAD detector and a Waters H-Class UPLC with a PDA detector under the conditions shown in Table 6.

[0167] Table 6

[0168]

[0169] Approximately 10 mg of the gentisate form 2 of compound 1 (calculated as the weight of the free base of compound 1) was placed into 3-mL glass vials and 1 mL of the corresponding medium (water, SGF, FaSSIF, and FeSSIF) was added to each glass vial. The lids were placed on each vial and they were rolled (25 rpm) at 37 °C for 4 hours. The suspension was then extracted into centrifuge tubes and then centrifuged (10000 rpm, 37 °C, 5 min) and filtered (0.22-μm PTFE membrane). The resulting supernatant was analyzed by HPLC and the pH was determined. The solubility of the polymorphic forms of the gentisate form 2 of compound 1 measured as described above is shown in Table 7.

[0170] Table 7

[0171]

[0172] Example 9: Tablets Containing the Gentisate Form of Compound 1

[0173] Tablets containing 261 mg of the gentisate form 2 of compound 1 (200 mg of the free base of compound 1) were prepared according to the following method. The amounts of the respective components used are shown in Table 8, and each component except magnesium stearate and sodium stearyl fumarate was passed through a #20 mesh sieve before use. Magnesium stearate and sodium stearyl fumarate were passed through a #35 mesh sieve before use.

[0174] Table 8

[0175]

[0176]

[0177] Put one-third of the sieved microcrystalline cellulose into a 10-liter hopper and blend it at 20 rpm for 5 minutes. Then add another one-third of the microcrystalline cellulose, the gentisate form of Compound 1, and the remaining one-third of the microcrystalline cellulose (for rinsing the bag containing the gentisate form of Compound 1), the first part of the cross-linked polyvinylpyrrolidone Kollidon CL, the first part of the colloidal silicon dioxide Aerosil 200 Pharma, and the first part of the sodium stearyl fumarate to the hopper. Then blend the resulting mixture at 20 rpm for 20 minutes. Pass the blended mixture through a #20 mesh sieve and add the sieved mixture to a 10-liter hopper and blend it further at 20 rpm for 15 minutes. Then add the first part of the magnesium stearate to the center of the blended mixture and blend it further at 20 rpm for 5 minutes. Then put the resulting mixture into a low-density polyethylene bag.

[0178] The roller press is equipped with the following settings: (a) roller width: 40 mm; (b) upper roller surface: knurled; (c) lower roller surface: knurled; (d) coarse RFG sieve: 2.0 mm, wire-wound; (e) fine RFG sieve: 1.0 mm, wire-wound. Then load the blended mixture from above into the feed hopper of the roller press and process the material using the parameters of the roller press shown in Table 9. Collect the granules (1454.25 g) into a low-density polyethylene bag.

[0179] Table 9

[0180] Parameter Setting Roll pressure (MPa) 1.8 Roll gap (mm) 3 Roll speed (rpm) 4 Granulator (RFG) speed (rpm) Fixed Feed screw speed (rpm) 30 Gap control Off Uncompacted fines Circulation Encapsulation density 1.0046 g / mL Solid fraction 0.653

[0181] Put the granules (1454.25 g) produced by the roller press into a 10-liter hopper, add sodium stearyl fumarate (7.725 g), colloidal silicon dioxide Aerosil 200 Pharma (7.78 g), and cross-linked polyvinylpyrrolidone Kollidon CL (22.503 g) to it, and blend the resulting mixture at 20 rpm for 15 minutes. Add magnesium stearate (7.72 g) to the blended mixture and blend the mixture at 20 rpm for 5 minutes to obtain the final blend.

[0182] The tablet press is equipped with the following: (a) upper punch, lower punch, and die head = 19 mm × 8.51 mm; (b) number of punches = 2; and (c) filling cam size = 8 mm to 14 mm. The tablet press is set with the parameters shown in Table 10 to obtain tablets within the target parameters shown in Table 11.

[0183] Table 10

[0184] Compression parameter Setting Rotation speed (rpm) 20 Feed speed (rpm) 27 Main compression (kN) 11.7 Fill depth (mm) 8.7

[0185] Table 11

[0186] Target parameter Target value Single tablet weight (mg) 663 to 714 Tablet thickness 5.4 mm to 5.5 mm Tablet hardness 170 N to 220 N Tablet friability Not exceeding 1% Disintegration time Less than 15 minutes

[0187] Example 10: Preparation of Form 2 of the Gentisate Salt of Compound 1

[0188] At about 25 °C under nitrogen, 2.8 kg of a 2:98 water / n-propanol mixture and 1.73 kg of the trifluoroacetic acid form of Compound 1 were placed into a reactor. The resulting mixture was stirred at a temperature of about 25 °C until the solid dissolved, then 1.31 kg of 2,5-dihydroxybenzoic acid (1.31 kg) was added, followed by an additional portion of the 2:98 water / n-propanol mixture. The resulting mixture was stirred at about 25 °C until the solid dissolved, then a portion of Form 2 of the gentisate salt of Compound 1 (about 0.02 kg) was added to the mixture, and the mixture was stirred at about 25 °C for an additional period of about 30 minutes. n-Heptane (about 22 kg) was added to the resulting mixture and the resulting mixture was stirred at about 25 °C for an additional period of about 16 hours. The resulting solid was filtered, washed and dried in vacuo in an oven with the temperature set at about 40 °C to about 50 °C and the relative humidity set at about 30% to about 40% to afford Form 2 of the gentisate salt of Compound 1 (about 2 kg).

[0189] Example 11: X-ray Powder Diffraction (XRPD) Analysis of Form 2 of the Gentisate Salt of Compound 1

[0190] A sample of Form 2 of the gentisate salt of Compound 1 prepared using a method similar to the method described in Example 10 was analyzed by XRPD and exhibited the peaks shown in Table 12. The error associated with each °2θ position was determined to be ±0.2°θ.

[0191] Table 12

[0192]

[0193] Example 12: Stability of Form 2 of the Gentisate Salt of Compound 1

[0194] To measure the stability of Form 2 of the gentisate salt form of Compound 1 under storage conditions, samples of Form 2 of the gentisate salt form of Compound 1 were placed in two low-density polyethylene bags with a desiccant between the bags, and each bag was placed in a high-density polyethylene drum. One drum was stored at a temperature of 5 °C and material samples were taken at 1 month and 3 months, and the samples were analyzed for the presence of impurities. The other drum was stored at a temperature of 25 °C and a relative humidity (RH) of 60%, material samples were taken at 1 month and 3 months, and the samples were analyzed for the presence of impurities. Using the set of test conditions and solvent gradients shown in Tables 13 and 14, the amount of Form 2 of the gentisate salt form of Compound 1 and the amount of any impurities in each sample were measured by reverse-phase high performance liquid chromatography. The samples were tested to determine the amount of Form 2 remaining at each time point and were measured by X-ray powder diffraction (XRPD) according to USP <941>.

[0195] Table 13

[0196] Table 14

[0197] Time (min) Mobile phase A % Mobile phase B % Initial 90 10 10 80 20 13 80 20 30 65 35 40 0 100 45 0 100 45.10 90 10 50 90 10

[0198] The results of the stability tests of Form 2 of the gentisate salt form of Compound 1 under the two storage conditions are shown in Table 15.

[0199] Table 15

[0200]

[0201] The results indicate that Form 2 of the gentisate salt form of Compound 1 is stable for up to 3 months when stored at 5 °C and is stable for up to 3 months when stored at 25 °C and 60% RH.

[0202] Embodiments

[0203] Embodiment 1: A pharmaceutically acceptable form of Compound 1:

[0204]

[0205] wherein the pharmaceutically acceptable form is selected from the gentisate salt form and the succinate salt form.

[0206] Embodiment 2: The pharmaceutically acceptable form according to Embodiment 1, wherein the pharmaceutically acceptable form is the gentisate salt form.

[0207] Embodiment 3: The pharmaceutically acceptable form according to Embodiment 2, wherein the gentisate salt form comprises gentisate.

[0208] Embodiment 4: A pharmaceutically acceptable form as described in Embodiment 2, wherein the gentisate form comprises a gentisate cocrystal.

[0209] Embodiment 5: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 4, wherein in the gentisate form, the molar ratio between Compound 1 and gentisic acid is about 1:1.

[0210] Embodiment 6: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 5, wherein the gentisate form is a hydrate.

[0211] Embodiment 7: A pharmaceutically acceptable form as described in Embodiment 6, wherein the hydrate is selected from hemihydrate, monohydrate and dihydrate.

[0212] Embodiment 8: A pharmaceutically acceptable form as described in Embodiment 7, wherein the hydrate is hemihydrate.

[0213] Embodiment 9: A pharmaceutically acceptable form as described in Embodiment 7, wherein the hydrate is monohydrate.

[0214] Embodiment 10: A pharmaceutically acceptable form as described in Embodiment 7, wherein the hydrate is dihydrate.

[0215] Embodiment 11: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 10, wherein the pharmaceutically acceptable form is a solid.

[0216] Embodiment 12: A pharmaceutically acceptable form as described in Embodiment 11, wherein the solid is a crystalline solid.

[0217] Embodiment 13: A pharmaceutically acceptable form as described in Embodiment 12, wherein the crystalline solid exhibits a peak at 9.25 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0218] Embodiment 14: A pharmaceutically acceptable form as described in Embodiment 13, wherein the crystalline solid exhibits further peaks at 6.97 ± 0.2° 2-θ, 20.53 ± 0.2° 2-θ and 26.08 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0219] Embodiment 15: A pharmaceutically acceptable form as described in Embodiment 13 or 14, wherein the crystalline solid exhibits further peaks at 14.61 ± 0.2° 2-θ and 18.89 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0220] Embodiment 16: A pharmaceutically acceptable form as described in any one of Embodiments 12 to 15, wherein the crystalline solid further comprises a peak at about 150 °C to about 170 °C in a differential scanning calorimetry profile.

[0221] Embodiment 17: A pharmaceutically acceptable form as described in Embodiment 16, wherein the crystalline solid comprises a peak at about 150 °C to about 165 °C in a differential scanning calorimetry profile.

[0222] Embodiment 18: A pharmaceutically acceptable form as described in Embodiment 16, wherein the crystalline solid comprises a peak at about 161 °C to about 162 °C in a differential scanning calorimetry profile.

[0223] Embodiment 19: A pharmaceutically acceptable form as described in any one of Embodiments 12 to 18, wherein the crystalline solid exhibits a mass loss of between about 1% and about 5% in thermogravimetric analysis when heated from about 31 °C to about 150 °C.

[0224] Embodiment 20: A pharmaceutically acceptable form as described in Embodiment 19, wherein the crystalline solid further exhibits a mass loss of between about 3% and about 5% in thermogravimetric analysis when heated from about 31 °C to about 150 °C.

[0225] Embodiment 21: A pharmaceutically acceptable form as described in Embodiment 19, wherein the crystalline solid further exhibits a mass loss of about 5% in thermogravimetric analysis when heated from about 31 °C to about 150 °C.

[0226] Embodiment 22: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 21, wherein the pharmaceutically acceptable form exhibits a solubility of at least 5 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37 °C.

[0227] Embodiment 23: A pharmaceutically acceptable form as described in Embodiment 22, wherein the pharmaceutically acceptable form exhibits a solubility of at least 10 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37 °C.

[0228] Embodiment 24: A pharmaceutically acceptable form as described in Embodiment 23, wherein the pharmaceutically acceptable form exhibits a solubility of at least 15 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37 °C.

[0229] Embodiment 25: A pharmaceutically acceptable form as described in Embodiment 23, wherein the pharmaceutically acceptable form exhibits a solubility of at least 20 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37 °C.

[0230] Embodiment 26: A pharmaceutically acceptable form as described in Embodiment 23, wherein the pharmaceutically acceptable form exhibits a solubility of at least 25 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37°C.

[0231] Embodiment 27: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 21, wherein the pharmaceutically acceptable form exhibits a solubility between about 10 mg / mL and about 30 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37°C.

[0232] Embodiment 28: A pharmaceutically acceptable form as described in Embodiment 27, wherein the pharmaceutically acceptable form exhibits a solubility between about 15 mg / mL and about 30 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37°C.

[0233] Embodiment 29: A pharmaceutically acceptable form as described in Embodiment 27, wherein the pharmaceutically acceptable form exhibits a solubility between about 20 mg / mL and about 30 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37°C.

[0234] Embodiment 30: A pharmaceutically acceptable form as described in Embodiment 27, wherein the pharmaceutically acceptable form exhibits a solubility between about 25 mg / mL and about 30 mg / mL in an aqueous solution at pH 1.7 and a temperature of 37°C.

[0235] Embodiment 31: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 30, wherein the pharmaceutically acceptable form exhibits a solubility of at least 5 mg / mL in an aqueous solution at pH 2.5 and a temperature of 37°C.

[0236] Embodiment 32: A pharmaceutically acceptable form as described in Embodiment 31, wherein the pharmaceutically acceptable form exhibits a solubility of at least 7.5 mg / mL in an aqueous solution at pH 2.5 and a temperature of 37°C.

[0237] Embodiment 33: A pharmaceutically acceptable form as described in Embodiment 31, wherein the pharmaceutically acceptable form exhibits a solubility of at least 10 mg / mL in an aqueous solution at pH 2.5 and a temperature of 37°C.

[0238] Embodiment 34: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 33, wherein the pharmaceutically acceptable form exhibits a solubility of at least 1 mg / mL in an aqueous solution at pH 4.4 and a temperature of 37°C.

[0239] Embodiment 35: A pharmaceutically acceptable form as described in Embodiment 34, wherein the pharmaceutically acceptable form exhibits a solubility of at least 2.5 mg / mL in an aqueous solution at pH 4.4 and a temperature of 37°C.

[0240] Embodiment 36: A pharmaceutically acceptable form as described in Embodiment 34, wherein the pharmaceutically acceptable form exhibits a solubility of at least 5 mg / mL in an aqueous solution at pH 4.4 and a temperature of 37°C.

[0241] Embodiment 37: A pharmaceutically acceptable form as described in Embodiment 34, wherein the pharmaceutically acceptable form exhibits a solubility of at least 7.5 mg / mL in an aqueous solution at pH 4.4 and a temperature of 37°C.

[0242] Embodiment 38: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 37, wherein when the pharmaceutically acceptable form is stored at 25°C and 60% relative humidity for at least 7 days, the pharmaceutically acceptable form exhibits less than about 10% degradation.

[0243] Embodiment 39: A pharmaceutically acceptable form as described in Embodiment 38, wherein when the pharmaceutically acceptable form is stored at 25°C and 60% relative humidity for at least 7 days, the pharmaceutically acceptable form exhibits less than about 1% degradation.

[0244] Embodiment 40: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 39, wherein when the pharmaceutically acceptable form is stored at 40°C and 75% relative humidity for at least 7 days, the pharmaceutically acceptable form exhibits less than about 10% degradation.

[0245] Embodiment 41: A pharmaceutically acceptable form as described in Embodiment 40, wherein when the pharmaceutically acceptable form is stored at 40°C and 75% relative humidity for at least 7 days, the pharmaceutically acceptable form exhibits less than about 1% degradation.

[0246] Embodiment 42: A pharmaceutically acceptable form as described in any one of Embodiments 1 to 41, wherein when the pharmaceutically acceptable form is stored at 60°C for one or more days, the pharmaceutically acceptable form exhibits less than about 10% degradation.

[0247] Embodiment 43: A pharmaceutically acceptable form as described in Embodiment 42, wherein when the pharmaceutically acceptable form is stored at 60°C for one or more days, the pharmaceutically acceptable form exhibits less than about 1% degradation.

[0248] Embodiment 44: A pharmaceutically acceptable form as described in Embodiment 12, wherein the crystalline solid exhibits a peak at 8.26 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0249] Embodiment 45: A pharmaceutically acceptable form as described in Embodiment 44, wherein the crystalline solid exhibits a further peak at 26.43 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0250] Embodiment 46: A pharmaceutically acceptable form as described in Embodiment 44 or 45, wherein the crystalline solid exhibits further peaks at 15.81 ± 0.2° 2-θ and 15.40 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0251] Embodiment 47: A pharmaceutically acceptable form as described in any one of Embodiments 44 to 46, wherein the crystalline solid exhibits further peaks at 14.94 ± 0.2° 2-θ and 20.44 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

[0252] Embodiment 48: A pharmaceutically acceptable form as described in Embodiment 1, wherein the pharmaceutically acceptable form is the succinate form.

[0253] Embodiment 49: A pharmaceutically acceptable form as described in Embodiment 48, wherein the succinate form contains succinate.

[0254] Embodiment 50: A pharmaceutically acceptable form as described in Embodiment 48, wherein the succinate form contains a succinate cocrystal.

[0255] Embodiment 51: A pharmaceutical composition comprising an amount of a pharmaceutically acceptable form of Compound 1 as described in any one of Embodiments 1 to 50, and one or more pharmaceutically acceptable excipients.

[0256] Embodiment 52: A method of inhibiting CD73 activity in a cell, which comprises contacting CD73 in the cell with an effective amount of a pharmaceutically acceptable form as described in any one of Embodiments 1 to 50.

[0257] Embodiment 53: A method of treating cancer in a subject, which comprises administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form as described in any one of Embodiments 1 to 50.

[0258] Embodiment 54: A method of treating cancer in a subject, which comprises administering to the subject the pharmaceutical composition as described in Embodiment 51.

[0259] Embodiment 55: The method according to embodiment 53 or 54, wherein the cancer is lung cancer, melanoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, prostate cancer, kidney cancer, multiple myeloma or lymphoma.

[0260] Embodiment 56: The method according to any one of embodiments 53 to 55, wherein the cancer expresses CD73.

[0261] Embodiment 57: The method according to any one of embodiments 53 to 56, wherein CD73 is upregulated in the cancer to be treated.

[0262] Embodiment 58: The method according to any one of embodiments 52 to 57, further comprising administering one or more second therapeutic agents.

[0263] Embodiment 59: The method according to embodiment 58, wherein the second therapeutic agent is a chemotherapeutic agent or an immunotherapeutic agent.

[0264] Embodiment 60: A method of treating an infection in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable form according to any one of embodiments 1 to 50.

[0265] Embodiment 61: A method of treating an infection in a subject, comprising administering to the subject a pharmaceutical composition according to embodiment 51.

[0266] Embodiment 62: The method according to embodiment 60 or 61, wherein the infection is a viral infection.

[0267] Embodiment 63: The method according to embodiment 60 or 61, wherein the infection is a parasitic infection.

[0268] Embodiment 64: A method of treating a neurodegenerative disease in a subject, comprising administering to the subject a pharmaceutically acceptable form according to any one of embodiments 1 to 50.

[0269] Embodiment 65: A method of treating a neurodegenerative disease in a subject, comprising administering to the subject a pharmaceutical composition according to embodiment 51.

[0270] Embodiment 66: The method according to embodiment 64 or 65, wherein the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, Huntington's disease, schizophrenia or autism.

[0271] Although the preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Many variations, changes, and substitutions will now occur to those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the present invention. The following claims are intended to define the scope of the present invention and thereby cover methods and structures within the scope of these claims and their equivalents.

Claims

1. A pharmaceutically acceptable form of Compound 1: wherein the pharmaceutically acceptable form is selected from the gentisate form and the succinate form.

2. The pharmaceutically acceptable form according to claim 1, wherein the pharmaceutically acceptable form is the gentisate form.

3. The pharmaceutically acceptable form according to claim 2, wherein in the gentisate form, the molar ratio between Compound 1 and gentisic acid is about 1:

1.

4. The pharmaceutically acceptable form according to claim 3, wherein the gentisate form is a crystalline solid.

5. The pharmaceutically acceptable form according to claim 4, wherein the crystalline solid exhibits a peak at 9.25 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

6. The pharmaceutically acceptable form according to claim 5, wherein the crystalline solid exhibits further peaks at 6.97 ± 0.2° 2-θ, 20.53 ± 0.2° 2-θ and 26.08 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

7. The pharmaceutically acceptable form according to claim 6, wherein the crystalline solid exhibits further peaks at 14.61 ± 0.2° 2-θ and 18.89 ± 0.2° 2-θ in the X-ray powder diffraction (XRPD) pattern.

8. The pharmaceutically acceptable form according to any one of claims 5 to 7, wherein the crystalline solid further comprises a peak at about 150 °C to about 170 °C in the differential scanning calorimetry pattern.

9. The pharmaceutically acceptable form according to any one of claims 1 to 8, wherein when the pharmaceutically acceptable form is stored at 25 °C and 60% relative humidity for at least 7 days, the pharmaceutically acceptable form exhibits less than about 10% degradation.

10. The pharmaceutically acceptable form according to claim 1, wherein the pharmaceutically acceptable form is the succinate form.

11. A pharmaceutical composition comprising an amount of the pharmaceutically acceptable form of Compound 1 according to any one of claims 1 to 10, and one or more pharmaceutically acceptable excipients.

12. A method of inhibiting CD73 activity in a cell, which comprises contacting CD73 in the cell with an effective amount of the pharmaceutically acceptable form according to any one of claims 1 to 10.

13. A method of treating cancer in a subject, which comprises administering to the subject a therapeutically effective amount of the pharmaceutically acceptable form according to any one of claims 1 to 10.

14. A method of treating cancer in a subject, which comprises administering to the subject the pharmaceutical composition according to claim 11.

15. The method according to claim 13 or 14, wherein the cancer is lung cancer, melanoma, breast cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, prostate cancer, kidney cancer, multiple myeloma or lymphoma.

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