Crystalline form of CD73 inhibitors and their use

Abstract

To provide a crystalline form of a compound of formula (I) that modulates the conversion of AMP to adenosine by 5'-nucleotidase, a composition containing the compound, and a method for preparing the crystalline form. [Solution] The use of such crystalline forms and compositions of formula (I) for the treatment and / or prevention of a variety of diseases, disorders, and conditions mediated by 5'-nucleotidase, including cancer and immune-related disorders, is also provided. This disclosure relates to crystalline forms of compounds that modulate the conversion of AMP to adenosine by ecto-5'-nucleotidase (also known as NT5E or 5NT, CD73), and compositions (e.g., pharmaceutical compositions) comprising such compounds.

Description

[Technical Field]

[0001] Cross-reference of related applications This application asserts the interests of U.S. Provisional Patent Application No. 63 / 040,277, filed on 17 June 2020, under 119(e) of the U.S. Patent Act, which is incorporated herein by reference in its entirety for all purposes.

[0002] For example, crystalline forms and compositions of compounds for the inhibition of adenosine by ecto-5'-nucleotidase, also known as CD73, and pharmaceutical compositions containing the same are provided herein. Also provided herein, for example, are methods for treating or preventing diseases, disorders, conditions, or symptoms thereof mediated by the inhibition of adenosine by ecto-5'-nucleotidase. [Background technology]

[0003] Ectonucleotides catalyze the conversion of ATP to adenosine, an endogenous regulator that affects multiple systems, including the immune, cardiovascular, central nervous, and respiratory systems. Adenosine also promotes fibrosis in various tissues. In the first step of adenosine production, ectonucleoside triphosphate diphosphohydrolase 1, also known as CD39 (cluster 39 of differentiation), is involved. ENTPD1 hydrolyzes ATP to ADP, and then hydrolyzes ADP to AMP. In the next step, AMP is converted to adenosine by ecto-5'-nucleotidase (NT5E or 5NT), also known as CD73 (cluster 73 of differentiation).

[0004] The enzymatic activities of CD39 and CD73 play a strategic role in calibrating the duration, scale, and chemical nature of purinergic signals delivered to various cells (e.g., immune cells). Alterations in these enzymatic activities can change the course or determine the prognosis of several pathophysiological events, including cancer, autoimmune diseases, infections, atherosclerosis, and ischemia-reperfusion injury, suggesting that these ectoenzymes represent novel therapeutic targets for managing various disorders.

[0005] Inhibition of CD73 by monoclonal antibodies, siRNA, or small molecules delays tumor growth and metastasis (Stagg, J. (2010) PNAS U.S.A. 107:1547 - 52). For example, anti-CD73 antibody therapy has been shown to inhibit the growth and metastasis of breast tumors in animal models (Stagg, J. (26 Jan 2010) PNAS U.S.A, 107(4):1547 - 52). Furthermore, the use of antibodies that specifically bind to CD73 has been evaluated for the treatment of bleeding disorders (e.g., hemophilia) (U.S. Patent No. 9,090,697). Several attempts have been made to develop therapeutically useful small molecule inhibitors of CD73. However, the development of small molecules has been hampered, for example, by suboptimal physical and metabolic stability. The compound designated as Compound I herein, ((((2R,3S,4R,5R)-5-(6-chloro-4-(((S)-1-(2-fluorophenyl)ethyl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)-phosphoryl)methyl)phosphonic acid, is a potent and selective small molecule inhibitor of CD73. Given the role played by CD73 in cancer, as well as in a variety of other diseases, disorders, and conditions, and the lack of CD73 inhibitors currently available to medical practitioners, there is a need for stable crystalline forms of Compound I, as well as related compositions and methods.

Prior Art Documents

Non-Patent Documents

[0006]

Non - Patent Document 1

Summary of the Invention

Means for Solving the Problems

[0007] The present disclosure relates to crystalline forms of compounds that modulate the conversion of AMP to adenosine by ecto 5'-nucleotidase (NT5E or 5NT, also known as CD73), and to compositions (e.g., pharmaceutical compositions) containing such compounds. Such compounds (crystalline forms), including methods of preparation, methods of use, and compositions, are described in detail below.

[0008] [[ID=严20]]In one aspect, the present disclosure provides a compound having the formula (I):

[0009]

Chemical Formula

[0010] In another aspect, the present disclosure provides a process for preparing a crystalline form of a compound of formula (I), the process comprising a) forming a first mixture comprising a compound of formula (I) and a solvent at a temperature of at least 20°C; b) adding an antisolvent to the first mixture to form a second mixture; or a) forming a first mixture comprising a compound of formula (I) and a solvent at a temperature of at least 20°C; c) cooling and stirring the first or second mixture to form a precipitate. d) isolating the precipitate; e) drying the precipitate to obtain the crystalline form of formula (I), comprising: where the solvent is C 1~4 alkyl alcohol, di-(C 1~4 alkyl) ether, 5-6 membered cyclic ether, acetic acid, or water, and the poor solvent is C 5~7 alkane, C 1~4 alkyl alcohol, di-(C 1~4 alkyl) ether, 5-6 membered cyclic ether, di-(C 1~4 alkyl) ketone, C 1~4 alkyl-C(O)O-C 1~4 alkyl, or aromatic hydrocarbon solvent, provided that the solvent and the poor solvent are each not C alkyl alcohol, di-(C 1~4 alkyl) ether, or 5-6 membered cyclic ether, a process is provided. 1~4

[0011] The present disclosure also relates to the use of such crystalline forms and compositions of such compounds for the treatment and / or prevention of various diseases, disorders, and conditions mediated in whole or in part by CD73. CD73 inhibitors have been associated with the treatment of a variety of disorders, including cancer, fibrosis, neurological and neurodegenerative disorders (e.g., depression and Parkinson's disease), cerebral and cardiac ischemic diseases, immune-related disorders, and disorders having an inflammatory component. [See, e.g., Sorrentino et al (2013) OncoImmunol, 2:e22448, doi:10.4161 / onci.22448; and Regateiro et al. (2012) Clin..Exp.Immunol, 171:1-7]. In certain embodiments, the crystalline forms of the compounds described herein can be formulated in a manner that inhibits the immunosuppressive and / or anti-inflammatory activity of CD73 and are useful as a therapeutic or prophylactic treatment when such inhibition is desired. Unless otherwise indicated, when referring to the compounds (or crystalline forms of the compounds) described herein, it should be understood that such compounds can be in a form suitable for delivery (e.g., a pharmaceutical composition).​

[0012] In some embodiments, the Disclosure envisions a method for treating or preventing cancer in a subject (e.g., a human), comprising administering to the subject a therapeutically effective amount of a crystalline form of the compound of formula (I). The Disclosure includes a method for treating or preventing cancer in a subject by administering to the subject a crystalline form of the compound of formula (I) in an amount effective in reversing, halting, or delaying the progression of CD73-mediated immunosuppression.

[0013] Examples of cancers that can be treated with the crystalline forms and compositions of the compound of formula (I) described herein include, but are not limited to, cancers of the prostate, e.g., metastatic castration-resistant prostate cancer; cancers of the colon, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovaries, testes, head, neck, skin (including melanoma and basal cell carcinoma), mesothelial layer, leukocytes (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell lung cancer and non-small cell lung cancer), adrenal gland, thyroid, kidney, or bone; glioblastoma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma, choriocarcinoma, cutaneous basal cell carcinoma; and testicular seminoma. In some embodiments of this disclosure, the cancer is melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia, brain tumor, lymphoma, sarcoma, ovarian cancer, or Kaposi's sarcoma. The crystalline forms and compositions of the compound of formula (I) of this disclosure are candidate cancers for treatment and are discussed further below.

[0014] This disclosure envisions a method for treating subjects undergoing bone marrow transplantation or peripheral blood stem cell transplantation by administering a therapeutically effective amount of a crystalline form of the compound of formula (I) sufficient to increase delayed-type hypersensitivity responses to tumor antigens, delay the time to recurrence of malignant tumors after transplantation, increase recurrence-free survival after transplantation, and / or increase long-term post-transplant survival.

[0015] In certain embodiments, the Disclosure envisions a method for treating or preventing an infectious disorder (e.g., a viral infection) in a subject (e.g., a human), comprising administering to the subject a therapeutically effective amount of a compound of formula (I) in crystalline form. In some embodiments, the infectious disorder is a viral infection (e.g., a chronic viral infection), a bacterial infection, a fungal infection, or a parasitic infection. In certain embodiments, the viral infection is human immunodeficiency virus or cytomegalovirus.

[0016] In further embodiments, the disclosure envisions methods for treating and / or preventing immune-related diseases, disorders, and conditions, diseases having inflammatory components, and disorders associated therewith, using the crystalline form of the compound of formula (I). Examples of immune-related diseases, disorders, and conditions are described below.

[0017] Other diseases, disorders, and conditions that can be treated or prevented, in whole or in part, by modulation of CD73 activity are candidate indications for the crystalline form of the compound of formula (I) described herein.

[0018] This disclosure further envisions the use of the crystalline form of the compound of formula (I) described herein in combination with one or more additional agents. The one or more additional agents may have some CD73 modulating activity and / or they may function through distinct mechanisms of action. In some embodiments, such agents include radiation (e.g., local or total radiotherapy) and / or other therapeutic modalities of non-pharmacological nature. When combination therapy is utilized, the crystalline form of the compound of formula (I) and one additional agent may be in the form of a single composition or multiple compositions, and the therapeutic modalities may be administered simultaneously, sequentially, or through several other regimens. As an example, this disclosure envisions a therapeutic regimen in which a chemotherapy phase follows an radiation phase. Combination therapy may have additive or synergistic effects. Other advantages of combination therapy are described below. [Brief explanation of the drawing]

[0019] [Figure 1] The X-ray powder diffraction (XRPD) pattern of crystalline form I of the compound of formula (I) is shown. [Figure 2] The differential scanning calorimetry (DSC) plot of crystalline form I of the compound of formula (I) is shown. [Figure 3] The X-ray powder diffraction (XRPD) pattern of crystalline form II of the compound of formula (I) is shown. [Figure 4] The differential scanning calorimetry (DSC) plot of the crystalline form II of the compound of formula (I) is shown. [Figure 5] The X-ray powder diffraction (XRPD) pattern of the compound of formula (I) in crystalline form III is shown. [Figure 6] The differential scanning calorimetry (DSC) plot of crystalline form III of the compound of formula (I) is shown. [Figure 7] The X-ray powder diffraction (XRPD) pattern of the compound of formula (I) in crystalline form IV is shown. [Figure 8] The X-ray powder diffraction (XRPD) pattern of the crystalline form V of the compound of formula (I) is shown. [Figure 9] The differential scanning calorimetry (DSC) plot of the crystalline form V of the compound of formula (I) is shown. [Figure 10] The X-ray powder diffraction (XRPD) pattern of crystalline form VI of the compound of formula (I) is shown. [Figure 11] The differential scanning calorimetry (DSC) plot of crystalline form VI of the compound of formula (I) is shown. [Figure 12a] The X-ray powder diffraction (XRPD) patterns of solids recovered from competitive slurry experiments using various amounts of ethanol and ethyl acetate are shown. The XRPD patterns of the reference material for Form I, the starting material for Form V, the reference material for Form II, and the material recovered by slurring mixtures of Forms I, II, and V in 100% EtOH at time 0 and 1 day, as well as the material recovered by slurring mixtures of Forms I, II, and V in 80% ethanol:20% ¼ at time 0, 1 day, and 2 days. [Figure 12b]The XRPD patterns of solids recovered from competitive slurry experiments using various amounts of ethanol and ethyl acetate are shown. The XRPD patterns of the reference material for Form I, the starting material for Form V, the reference material for Form II, and the material recovered by slurring mixtures of Forms I, II, and V in 67% EtOH:33% ¼ at time 0, 1 day, and 2 days, as well as the material recovered by slurring mixtures of Forms I, II, and V in 50% ethanol:50% ¼ at time 0 and 1 day, are shown. [Figure 12c] The X-ray powder diffraction (XRPD) patterns of solids recovered from competitive slurry experiments using varying amounts of ethanol and ethyl acetate are shown. The XRPD patterns of the reference material for Form I, the starting material for Form V, the reference material for Form II, and the material recovered by slurring mixtures of Forms I, II, and V in 33% EtOH:67% ¼ at time 0, 1 day, and 2 days are also shown. [Figure 12d] The XRPD patterns of solids recovered from competitive slurry experiments using varying amounts of ethanol and ethyl acetate are shown. The XRPD patterns of the reference material for Form I, the starting material for Form V, the reference material for Form II, and the material recovered by slurring mixtures of Forms I, II, and V in 20% EtOH:80% SiO2 at 0 hours and 2 days, as well as the material recovered by slurring mixtures of Forms I, II, and V in 100% SiO2 at 0 hours and 2 days, are also shown. [Figure 13] This shows the solubility profiles of shapes I (rhomboid), II (square), and V (triangular) in the EtOH / SiO solvent system at 20°C. [Figure 14] This shows a polymorphism relationship map based on observations from slurry experiments using an EtOH:SiO solvent system. The conditions shown in the relationship map are illustrative and are not intended to represent all possible pathways that can be used to produce the shown crystalline morphologies. [Figure 15]The solubility profiles for form I (square) at 20°C, form I (circular) at 35°C, a mixture of forms II and V (rhomboid) at 20°C, and a mixture of forms II and V (triangle) at 35°C are shown in the EtOH / heptane solvent system. [Figure 16] The dynamic vapor sorption (DVS) isotherm for form I of the compound of formula (I) is shown. [Figure 17] The dynamic vapor adsorption (DVS) isotherm for form II of the compound of formula (I) is shown. [Modes for carrying out the invention]

[0020] (Detailed description of the invention) Before further description of this disclosure, please understand that this disclosure is not limited to the specific embodiments described herein, and that the terms used herein are for the purpose of describing specific embodiments only and are not intended to limit them.

[0021] Where a range of values ​​is provided, it is understood that the respective intervening values ​​between the upper and lower limits of that range, up to one-tenth of the lower limit unit unless otherwise clearly indicated by the context, and any other mentioned or intervening values ​​within that range, are included in this disclosure. The upper and lower limits of these smaller ranges may independently be included in smaller ranges, are included in this disclosure, and are subject to any specifically excluded limits within the range referred to. If a range referred to includes one or both of the limit values, the range excluding one or both of those included limit values ​​is also included in this disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art in which this disclosure belongs.

[0022] Where used herein, the singular “a,” “an,” and “the” include multiple references unless the context clearly indicates otherwise. It should be further noted that claims may be constructed to exclude any element. Therefore, this statement is intended to function as an antecedent or “negative” restriction for the use of exclusive terms such as “solely” or “only” in relation to the enumeration of claim elements.

[0023] As used herein, the term “crystalline form of the compound of formula (I)” refers to any of the crystalline forms of the compounds mentioned herein. However, the crystalline forms may be formulated, for example, into liquids, gels, or ointments to facilitate administration to a subject. In particular, with respect to a method comprising the administration of the crystalline form of the compound of formula (I), the method means comprising the administration of a liquid formulation prepared using the crystalline form of the compound of formula (I).

[0024] The publications discussed herein are provided solely for disclosure prior to the filing date of this application. Furthermore, the dates of the provided publications may differ from the actual publication dates, which may require independent verification.

[0025] I. Overview The compound (((((2R,3S,4R,5R)-5-(6-chloro-4-(((S)-1-(2-fluorophenyl)ethyl)amino)-1H-pyrazolo[3,4-b]pyridine-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid, represented by formula (I), is a potent inhibitor of CD73.

[0026] [ka]

[0027] This disclosure arises from the remarkable discovery of the crystalline forms of the compound of formula (I), the advantages attributed to the forms described herein, and the processes for producing the crystalline forms. Crystalline materials are generally more physically and chemically stable. Since the shelf life of the product is directly correlated with stability, the superior stability of crystalline materials makes them more suitable for use in the final dosage form.

[0028] The manufacturing process of a compound can also play a role in selecting the desired polymorphic form. For example, compounds exhibiting transient solubility due to certain polymorphic forms tending to precipitate from solution may present challenges in maximizing the efficiency of the manufacturing protocol. In such cases, understanding the solubility profiles of the polymorphic forms of a compound can be crucial for process development. For instance, solubility profiles can provide deterministic information about solvent systems that can be used to solubilize all polymorphic forms of the desired compound, enabling purification techniques such as polishing filtration to minimize the loss of the desired compound due to precipitation on the filter. Following such purification techniques, the solubility profiles of the various polymorphic forms can provide deterministic information about techniques useful for precipitation of the desired compound from solution so that the overall yield is maximized (i.e., by controlling solvent conditions to induce precipitation of the most stable polymorphic form under those conditions). Certain polymorphic forms may also possess physical properties that make them easier to handle during the manufacturing process. For example, crystal morphology (e.g., needle-like, plate-like, or columnar) can affect the ease of filtration and drying protocols. Additional physical properties, such as hygroscopicity, bulk density, and fluidity, can offer certain advantages in the manufacturing process. Furthermore, the crystallization step in the processing of active pharmaceutical ingredients (APIs) also offers an opportunity to enhance the purity of the drug substance by removing impurities (e.g., those in the processing solvent). Finally, certain polymorphic forms may be selected due to their suitability for pharmaceutical applications, for example, having a residual solvent content that is safe for patient administration (e.g., orally or parenterally).

[0029] II. Definition Unless otherwise indicated, the following terms are intended to have the meanings set forth below. Other terms are defined elsewhere throughout this specification.

[0030] "Alkyl" has the indicated number of carbon atoms (i.e., C 1~4 A group can have 1 to 4 carbon atoms, and can be linear or branched, saturated, or aliphatic. An alkyl group can have any number of carbon atoms, for example, C 1~2 , C 1~3 , C 1~4 , C 2~3 , and C 3~4 It may include C. 1~4 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl.

[0031] A "hydrate" refers to a complex formed by combining the compound of formula (I) with water. This term includes both stoichiometric and non-stoichiometric hydrates.

[0032] A "solvate" refers to a complex formed by combining a compound of formula (I) with a solvent. This term includes both stoichiometric and non-stoichiometric solvates. Exemplary solvents that form solvates include, but are not limited to, methanol, ethanol, isopropanol, DMSO, ethyl acetate, acetic acid, acetonitrile, and methyl tert-butyl ether. In some embodiments, the crystalline form of the compound of formula (I) is acetonitrile, ethanol, ethyl acetate, or methyl tert-butyl ether solvate.

[0033] "Desolvated" refers to a form of the compound of formula (I) from which solvent molecules have been partially or completely removed, as described herein. Desolvation techniques for producing the desolvated form include, but are not limited to, exposure of the compound of formula (I) (solvate) to a vacuum, exposure of the solvate to a high temperature, exposure of the solvate to a gas stream such as air or nitrogen, washing or slurring of the solvate in a different solvent having a low binding tendency, or any combination thereof. Therefore, the desolvated form of the compound of formula (I) may not contain any solvent molecules at all, or it may be partially solvated with solvent molecules present in stoichiometric or non-stoichiometric amounts.

[0034] "Alcohol" refers to a solvent containing a hydroxyl group. Typical alcohols may have any suitable number of carbon atoms, such as C1 to C6, and any suitable number of hydroxyl groups, such as 1 to 3. Examples of alcohols include, but are not limited to, methanol, ethanol, n-propanol, and i-propanol.

[0035] "Crude" refers to a mixture containing the desired compound (e.g., the compound of formula (I)) and at least one other type (e.g., a solvent, a reagent, e.g., an acid or a base, a starting material, or a byproduct of a reaction that produces the desired compound).

[0036] The preferred solvents described herein refer to solvents characterized by high solubility of the compound of formula (I) at a concentration of at least about 50 mg / mL at 55–60°C. Poor solvents are generally considered "unsuitable solvents" and refer to solvents characterized by low solubility of the compound of formula (I) at a concentration of less than about 50 mg / mL at 55–60°C. Poor solvents may have poor solubility of the compound, but they may be perfectly suitable for crystallization purposes.

[0037] "Precipitation" refers to the process of combining compounds in a solution to form a solid substance (i.e., a precipitate). All or any fraction of a compound in a solution can be precipitated. The solid form of the substance can be amorphous or crystalline.

[0038] "Crystalline form" refers to the solid form of a compound in which constituent molecules are packed in a repeating pattern of regularly arranged structures. Crystalline forms can include triclinic, monoclinic, orthorhombic, tetragonal, trigonal, hexagonal, and cubic shapes. A crystalline form can include one or more regions, i.e., particles with distinct crystal boundaries. A crystalline solid can contain two or more crystal shapes.

[0039] "Amorphous form" refers to the solid form of a compound that does not have a clear crystalline structure, that is, it lacks a repeating pattern of regularly arranged constituent molecules.

[0040] "Isolation" refers to the process of isolating at least a portion of a first substance (e.g., a precipitate) from a mixture containing that substance and at least one additional substance. In some cases, the isolated substance substantially contains at least one of the additional substances present in the original mixture.

[0041] "Substantially absent" means an amount of another form or impurity of 10% or less, preferably 8%, 5%, 4%, 3%, 2%, 1%, 0.5%, or less. Preferably, substantially absent means a crystalline form of the compound of formula (I) containing less than 5% of another crystalline or amorphous form of the compound of formula (I). Preferably, substantially absent means a crystalline form of the compound of formula (I) containing less than 1% of another crystalline or amorphous form of the compound of formula (I).

[0042] "Approximately" means a range of values ​​that includes the specified value, which a person skilled in the art would reasonably consider to be similar to the specified value. In some embodiments, the term "approximately" means within a standard deviation using generally acceptable measurements in the art. In some embodiments, "approximately" means a range of up to ±10% of the specified value. In some embodiments, "approximately" means a range of up to ±5% of the specified value. In some embodiments, "approximately" means a range of up to ±2% of the specified value. In some embodiments, "approximately" means the specified value.

[0043] The term "pharmaceutically acceptable salt" means that, depending on the specific substituents found in the compounds described herein, salts of the active compound prepared using relatively non-toxic acids or bases may be included. If the compounds of this disclosure contain relatively acidic functional groups, base addition salts may be obtained by contacting the neutral form of such compound with a sufficient amount of the desired base, either undiluted or in a suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganese, manganese, potassium, sodium, and zinc. Examples of pharmaceutically acceptable salts derived from organic bases include substituted amines, cyclic amines, and naturally occurring amines, such as salts of primary, secondary, and tertiary amines including arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydravamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, and tromethamine. If the compounds of this disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either in an undiluted or suitable inert solvent.Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphoric acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids such as alginates, and salts of organic acids such as glucuronic acid or galacturonic acid are also included (e.g., Berge, SM, et al, "Pharmaceutical..."). See "Salts," Journal of Pharmaceutical Science, 1977, 66, 1-19. Certain specific compounds in this disclosure contain both basic and acidic functional groups that enable the conversion of the compound into either a base addition salt or an acid addition salt.

[0044] The neutral form of the compound can be regenerated by contacting the salt with a base or acid and isolating the parent compound by conventional means. The parent form of the compound has certain physical properties, such as solubility in polar solvents, that differ from the various salt forms, but in other respects, the salt is equivalent to the parent form of the compound for the purposes of this disclosure.

[0045] The compounds of this disclosure may also contain non-natural proportions of atomic isotopes in one or more of the atoms constituting such compounds. Non-natural proportions of isotopes can be defined as ranging from amounts found in nature to amounts comprising 100% of the atom in question. For example, a compound may contain, for example, tritium ( 3 H) or carbon-14 ( 14 Radioactive isotopes such as C, or deuterium ( 2 H) or carbon-13 ( 13Non-radioactive isotopes such as C) can be incorporated. Such isotopes of the compounds of this disclosure may find additional utility, including but not limited to diagnostic and / or imaging reagents, or cytotoxic / radiotoxic therapeutic agents. Furthermore, isotopic variants of the compounds of this disclosure may have altered pharmacokinetic and pharmacodynamic characteristics that may contribute to enhanced safety, tolerance, or efficacy during treatment. All isotopic variants of the compounds of this disclosure, whether radioactive or not, are intended to be included within the scope of this disclosure.

[0046] The terms "patient" or "subject" are used interchangeably to refer to human or non-human animals (e.g., mammals).

[0047] The terms "to treat," "to treat," and "treatment" refer to a process of action initiated, either temporarily or permanently, to eliminate, reduce, suppress, alleviate, or improve at least one of the underlying causes of a disease, disorder, or condition, or its symptoms, or at least one of the symptoms associated with the disease, disorder, or condition, after a diagnosis, observation, etc., of the disease, disorder, or condition, or its symptoms. Therefore, treatment includes inhibiting active disease (e.g., stopping the onset or further onset of a disease, disorder, or condition, or its associated clinical symptoms).

[0048] As used herein, the term “requiring treatment” refers to a judgment made by a physician or other caregiver that the subject requires treatment or would benefit from treatment. This judgment is based on various factors within the scope of the physician's or caregiver's expertise.

[0049] The terms “prevent,” “prevention,” and “prevention” generally refer to a process of action initiated (e.g., administering a CD73 inhibitor or a pharmaceutical composition containing one) that temporarily or permanently prevents, suppresses, inhibits, or reduces the risk of a subject developing a particular disease, disorder, or condition, or delays its onset (e.g., before the onset of the disease, disorder, condition, or its symptoms), given that the subject is predisposed to such a condition. In certain cases, these terms may also refer to delaying the progression of a disease, disorder, or condition, or inhibiting its progression to a harmful or otherwise undesirable condition.

[0050] As used herein, the term “requiring prevention” refers to a judgment made by a physician or other caregiver that a person requires or would benefit from preventive measures. This judgment is based on various factors that fall within the scope of the physician's or caregiver's expertise.

[0051] The term "therapeutic dose" refers to the amount of a drug administered to a subject that, when administered to the subject, can have any detectable positive effect on any symptom, aspect, or characteristic of a disease, disorder, or condition, whether alone or as part of a pharmaceutical composition, and whether as a single dose or as part of a series of doses. The therapeutic dose can be determined by measuring the relevant physiological effects, which can be adjusted in relation to the administration regimen and diagnostic analysis of the subject's condition, etc. For example, measuring serum levels of a CD73 inhibitor (or, for example, its metabolites) at a specific time point after administration may indicate whether a therapeutic dose has been used.

[0052] The phrase "enough to produce a change" means that there is a detectable difference between the level of an indicator measured before administration of a particular treatment (e.g., baseline level) and the level measured after administration. The indicator may include any objective parameter (e.g., serum concentration) or subjective parameter (e.g., the subject's perception of health).

[0053] "Substantially pure" means that the component makes up more than approximately 50% of the total composition, typically more than approximately 60% of the total composition. More typically, "substantially pure" refers to a composition in which the component of interest makes up at least 75%, at least 85%, at least 90%, or more of the total composition. In some cases, the component of interest may make up more than approximately 90%, or even more than approximately 95%, of the total composition.

[0054] As used herein, the terms “CD73 inhibitor,” “CD73 blocker,” “adenosine inhibitor by ecto-5'-nucleotidase,” “NT5E inhibitor,” “5NT inhibitor,” and all other terms permitted in the relevant art refer to compounds that can modulate the CD73 receptor, either directly or indirectly, in in vitro assays, in vivo models, and / or other means demonstrating therapeutic efficacy. The term also refers to compounds that demonstrate at least some therapeutic benefit in human subjects. CD73 inhibitors can be competitive, non-competitive, or irreversible CD73 inhibitors. A “competitive CD73 inhibitor” is a compound that reversibly inhibits CD73 enzyme activity at a catalytic site; a “non-competitive CD73 inhibitor” is a compound that reversibly inhibits CD73 enzyme activity at a non-catalytic site; and an “irreversible CD73 inhibitor” is a compound that irreversibly eliminates CD73 enzyme activity by forming a covalent bond with the enzyme (or by other stable means of inhibiting enzyme function).

[0055] III.Crystal form This disclosure provides crystalline forms of (((((2R,3S,4R,5R)-5-(6-chloro-4-(((S)-1-(2-fluorophenyl)ethyl)amino)-1H-pyrazolo[3,4-b]pyridine-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid represented by formula (I), including solvates and hydrates.

[0056] In one embodiment, this disclosure relates to formula (I):

[0057] [ka] A crystalline form of a compound having the following characteristics, wherein the crystalline form is one of crystalline forms I to VI, and each of these is diffracted by X-ray powder diffraction as described herein. It provides a crystalline morphology characterized by diffraction (XRPD) patterns.

[0058] Methods for acquiring XRPD data are known in the art, and any such method can be used to characterize the crystalline form of the compound of formula (I). For example, the X-ray powder diffraction pattern described herein can be generated using Cu Kα1 irradiation.

[0059] In some embodiments, the crystalline morphologies described herein are further characterized by differential scanning calorimetry (DSC) thermograms.

[0060] In some embodiments, the crystal morphology described herein has a nuclear magnetic resonance spectrum, 1 H Further characterization is performed by NMR spectroscopy.

[0061] In some embodiments, the crystalline morphologies described herein are further characterized by dynamic vapor adsorption (DVS) isotherms.

[0062] III-1.Crystal form I In one embodiment, the disclosure provides a crystalline form I of a compound of formula (I), characterized by an X-ray powder diffraction (XRPD) pattern having three or more peaks at 11.1, 11.6, 13.8, 14.7, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ).

[0063] The crystalline form I of the compound of formula (I) can be characterized by an X-ray powder diffraction (XRPD) pattern having one or more peaks at 11.1, 11.6, 13.8, 14.7, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ), for example, two, three, four, five, seven, or more peaks, where the XRPD is CuK α1It is fabricated using irradiation. In some embodiments, crystalline morphology I is characterized by an XRPD pattern containing four or more peaks at 11.1, 11.6, 13.8, 14.7, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline morphology I is characterized by an XRPD pattern containing five or more peaks at 11.1, 11.6, 13.8, 14.78, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline morphology I is characterized by an XRPD pattern containing six or more peaks at 11.1, 11.6, 13.8, 14.78, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline morphology I is characterized by an XRPD pattern containing seven or more peaks at 11.1, 11.6, 13.8, 14.78, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ).

[0064] In some embodiments, crystalline morphology I is characterized by an XRPD pattern containing peaks at 11.1, 13.8, 18.6, 20.1, 23.0, and 24.8 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes one or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes two or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes three or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes four or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes five or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ).

[0065] In some embodiments, crystalline morphology I is characterized by an XRPD pattern containing peaks at 11.1, 11.6, 13.8, 14.7, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline morphology I is characterized by an XRPD pattern containing peaks at 6.3, 8.0, 9.3, 11.1, 11.6, 13.8, 14.7, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, 29.1, 31.0, 31.9, and 33.4 degrees 2θ (±0.2 degrees 2θ).

[0066] In some embodiments, the crystal morphology I is characterized substantially by the XRPD pattern shown in Figure 1.

[0067] In some embodiments, crystalline form I substantially does not include other crystalline or amorphous forms of the compound of formula (I).

[0068] In some embodiments, crystalline morphology I is characterized by a differential scanning calorimetry (DSC) thermogram that includes endothermic activity between approximately 155°C and approximately 167°C. In some embodiments, crystalline morphology I is further characterized by a differential scanning calorimetry (DSC) thermogram that includes an endothermic peak at approximately 163.9°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram further includes exothermic activity between approximately 167°C and approximately 210°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram is characterized by an exothermic peak at approximately 176.1°C. In some embodiments, crystalline morphology I is further characterized by a melting point of approximately 163.9°C when determined by a differential scanning calorimetry (DSC) thermogram. In some embodiments, crystalline morphology I is characterized by a melting point of approximately 155.1°C when determined by a differential scanning calorimetry (DSC) thermogram. It is further characterized by its melting point onset.

[0069] In some embodiments, crystalline morphology I is further characterized by a differential scanning calorimetry (DSC) thermogram substantially as shown in Figure 2.

[0070] In some embodiments, the crystalline morphology I is characterized substantially by an XRPD pattern as shown in Figure 1 and further characterized substantially by a differential scanning calorimetry (DSC) thermogram as shown in Figure 2.

[0071] Crystal morphology I 1The content of one or more residual solvents (e.g., ethanol and / or toluene) can be determined using the 1H NMR spectrum. In some embodiments, the crystalline form I is 1 As determined by the 1H NMR spectrum, it contains 0.04 wt% ethanol. In some embodiments, crystalline form I is 1 When determined by 1H NMR spectroscopy, it is substantially free of toluene.

[0072] In some embodiments, crystalline morphology I is characterized by DVS isotherms characterized by a mass change of 0.5% to 6.5% at 40% RH to 70% RH. In some embodiments, crystalline morphology I is characterized by DVS isotherms characterized by a mass change of 2% to 5% at 50% RH to 60% RH. In some embodiments, crystalline morphology I is characterized substantially by DVS isotherms as shown in Figure 16.

[0073] III-2.Crystal form II In one embodiment, the disclosure provides a crystalline form II of a compound of formula (I), characterized by an X-ray powder diffraction (XRPD) pattern having three or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ).

[0074] The crystalline form II of the compound of formula (I) can be characterized by an X-ray powder diffraction (XRPD) pattern having one or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ), for example, two, three, four, five, seven, or more peaks, where the XRPD is CuK α1It is prepared using irradiation. In some embodiments, the crystalline form II of the compound of formula (I) is characterized by an XRPD pattern containing four or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form II of the compound of formula (I) is characterized by an XRPD pattern containing five or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form II of the compound of formula (I) is characterized by an XRPD pattern containing six or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form II of the compound of formula (I) is characterized by an XRPD pattern containing seven or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ).

[0075] In some embodiments, the crystalline form II of the compound of formula (I) is characterized by an XRPD pattern containing a peak at 16.5 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form II of the compound of formula (I) is characterized by an XRPD pattern containing peaks at 16.5, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes one or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes two or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes three or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes four or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes five or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ).

[0076] In some embodiments, crystalline morphology II is characterized by an XRPD pattern containing peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ).

[0077] In some embodiments, crystalline morphology II is characterized substantially by the XRPD pattern shown in Figure 3.

[0078] In some embodiments, crystalline form II substantially does not include other crystalline or amorphous forms of the compound of formula (I).

[0079] In some embodiments, crystalline morphology II is characterized by a differential scanning calorimetry (DSC) thermogram that includes endothermic activity between approximately 156°C and approximately 171°C. In some embodiments, crystalline morphology II is further characterized by a differential scanning calorimetry (DSC) thermogram that includes an endothermic peak at approximately 166.5°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram further includes exothermic activity between approximately 170°C and approximately 210°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram is characterized by an exothermic peak at approximately 179.3°C. In some embodiments, crystalline morphology II is further characterized by a melting point of approximately 166.5°C when determined by a differential scanning calorimetry (DSC) thermogram. In some embodiments, crystalline morphology II is further characterized by a melting point onset at approximately 157.4°C when determined by a differential scanning calorimetry (DSC) thermogram.

[0080] In some embodiments, crystalline morphology II is further characterized by a differential scanning calorimetry (DSC) thermogram substantially as shown in Figure 4.

[0081] In some embodiments, crystalline morphology II is characterized substantially by an XRPD pattern as shown in Figure 3, and further characterized substantially by a differential scanning calorimetry (DSC) thermogram as shown in Figure 4.

[0082] Crystallographic form II 1 The content of one or more residual solvents (e.g., ethanol) can be determined using the 1H NMR spectrum. In some embodiments, crystalline form II is 1 When determined by 1H NMR spectroscopy, it contains 0.68% by weight of ethanol.

[0083] In some embodiments, crystalline morphology II is characterized by DVS isotherms characterized by a mass change of 1% to 4% at 40% RH to 70% RH. In some embodiments, crystalline morphology II is characterized by DVS isotherms characterized by a mass change of 1.5% to 3.5% at 40% RH to 70% RH. In some embodiments, crystalline morphology II is characterized substantially by the DVS isotherms shown in Figure 17.

[0084] III-3.Crystal form III In one embodiment, the disclosure provides a crystalline form III of a compound of formula (I), characterized by an X-ray powder diffraction (XRPD) pattern having three or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ).

[0085] The crystalline form III of the compound of formula (I) can be characterized by an X-ray powder diffraction (XRPD) pattern having one or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ), for example, two, three, four, five, or more peaks, where the XRPD is CuK α1It is prepared using irradiation. In some embodiments, the crystalline form III of the compound of formula (I) is characterized by an XRPD pattern containing four or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form III of the compound of formula (I) is characterized by an XRPD pattern containing five or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form III of the compound of formula (I) is characterized by an XRPD pattern containing six or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form III of the compound of formula (I) is characterized by an XRPD pattern containing seven or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ).

[0086] In some embodiments, the crystalline form III of the compound of formula (I) is characterized by an XRPD pattern containing peaks at 6.6, 10.9, 14.2, 16.1, 18.4, and 19.3 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes one or more peaks at 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes two or more peaks at 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes three or more peaks at 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes four peaks at 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes peaks at 29.7, 32.0, and 33.5 degrees 2θ (±0.2 degrees 2θ).

[0087] In some embodiments, crystalline morphology III is characterized by an XRPD pattern containing peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline morphology III is characterized by an XRPD pattern containing peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, 28.1, 29.7, 32.0, and 33.5 degrees 2θ (±0.2 degrees 2θ).

[0088] In some embodiments, crystalline morphology III is substantially characterized by the XRPD pattern shown in Figure 5.

[0089] In some embodiments, crystalline form III substantially does not include other crystalline or amorphous forms of the compound of formula (I).

[0090] In some embodiments, crystalline morphology III is characterized by a differential scanning calorimetry (DSC) thermogram that includes endothermic activity between approximately 149°C and approximately 183°C. In some embodiments, crystalline morphology III is further characterized by a differential scanning calorimetry (DSC) thermogram that includes an endothermic peak at approximately 161.8°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram further includes exothermic activity between approximately 183°C and approximately 210°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram is characterized by an exothermic peak at approximately 188.2°C. In some embodiments, crystalline morphology III is further characterized by a melting point of approximately 161.8°C when determined by a differential scanning calorimetry (DSC) thermogram. In some embodiments, crystalline morphology III is further characterized by a melting point onset of approximately 149.6°C when determined by a differential scanning calorimetry (DSC) thermogram.

[0091] In some embodiments, crystalline morphology III is further characterized by a differential scanning calorimetry (DSC) thermogram substantially as shown in Figure 6.

[0092] In some embodiments, crystalline morphology III is characterized substantially by an XRPD pattern as shown in Figure 5, and further characterized substantially by a differential scanning calorimetry (DSC) thermogram as shown in Figure 6.

[0093] Crystallographic form III 1 The content of one or more residual solvents (e.g., ethanol and / or methyl tert-butyl ether) can be determined using the 1H NMR spectrum. In some embodiments, crystalline form III is 1 When determined by 1H NMR spectroscopy, it is substantially free of ethanol. In some embodiments, crystalline form III is 1 When determined by 1H NMR spectroscopy, it contains 0.40 wt% of methyl tert-butyl ether.

[0094] III-4.Crystal form IV In one embodiment, the disclosure provides a crystalline form IV of a compound of formula (I), characterized by an X-ray powder diffraction (XRPD) pattern having three or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ).

[0095] Crystal morphology IV of the compound of formula (I) can be characterized by an X-ray powder diffraction (XRPD) pattern having one or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ), for example, two, three, four, five, or more peaks, where the XRPD is CuK α1It is prepared using irradiation. In some embodiments, crystalline form IV of the compound of formula (I) is characterized by an XRPD pattern containing four or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline form IV of the compound of formula (I) is characterized by an XRPD pattern containing five or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline form IV of the compound of formula (I) is characterized by an XRPD pattern containing six or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline form IV of the compound of formula (I) is characterized by an XRPD pattern containing seven or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ).

[0096] In some embodiments, crystalline form IV of the compound of formula (I) is characterized by an XRPD pattern containing peaks at 14.1, 17.0, 19.5, 23.2, and 25.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes one or more peaks at 6.0, 11.2, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes two or more peaks at 6.0, 11.2, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes three or more peaks at 6.0, 11.2, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes four peaks at 6.0, 11.2, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ).

[0097] In some embodiments, crystalline morphology IV is characterized by an XRPD pattern containing peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ).

[0098] In some embodiments, crystalline morphology IV is substantially characterized by the XRPD pattern shown in Figure 7.

[0099] In some embodiments, crystalline form IV substantially does not include other crystalline or amorphous forms of the compound of formula (I).

[0100] Crystallographic form IV 1 The content of one or more residual solvents (e.g., THF and / or ethyl acetate) can be determined using the 1H NMR spectrum. In some embodiments, crystalline form IV is 1 When determined by 1H NMR spectroscopy, it has a THF content of 0.05%. In some embodiments, crystalline form IV is 1 When determined by 1H NMR spectroscopy, it contains 0.38% by weight of ethyl acetate.

[0101] III-5.Crystal form V In one embodiment, the disclosure provides a crystalline form V of a compound of formula (I), characterized by an X-ray powder diffraction (XRPD) pattern having three or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.5, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ).

[0102] The crystalline form V of the compound of formula (I) can be characterized by an X-ray powder diffraction (XRPD) pattern having one or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.6, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ), for example, two, three, four, five, or more peaks, where the XRPD is CuK α1 It is prepared using irradiation. In some embodiments, the crystalline form V of the compound of formula (I) is characterized by an XRPD pattern containing four or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.6, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form V of the compound of formula (I) is characterized by an XRPD pattern containing five or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.6, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form V of the compound of formula (I) is characterized by an XRPD pattern containing six or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.6, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form V of the compound of formula (I) is characterized by an XRPD pattern containing seven or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.6, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ).

[0103] In some embodiments, the crystalline form V of the compound of formula (I) is characterized by an XRPD pattern containing peaks at 15.8, 16.3, 16.8, 18.5, 19.1, 21.7, 22.1, and 23.0 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes one or more peaks at 10.4, 15.1, 19.7, and 23.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes two or more peaks at 10.4, 15.1, 19.7, and 23.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes three or more peaks at 10.4, 15.1, 19.7, and 23.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes four peaks at 10.4, 15.1, 19.7, and 23.6 degrees 2θ (±0.2 degrees 2θ).

[0104] In some embodiments, the crystal morphology V is characterized by an XRPD pattern containing peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, and 23.6 degrees 2θ (±0.2 degrees 2θ).

[0105] In some embodiments, the crystal morphology V is characterized substantially by the XRPD pattern shown in Figure 8.

[0106] In some embodiments, crystalline form V substantially does not include other crystalline or amorphous forms of the compound of formula (I).

[0107] In some embodiments, crystalline morphology V is characterized by a differential scanning calorimetry (DSC) thermogram that includes endothermic activity between approximately 135°C and approximately 172°C. In some embodiments, crystalline morphology V is further characterized by a differential scanning calorimetry (DSC) thermogram that includes an endothermic peak at approximately 150.4°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram further includes exothermic activity between approximately 171°C and approximately 210°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram is characterized by an exothermic peak at approximately 177.8°C. In some embodiments, crystalline morphology V is further characterized by a melting point of approximately 150.4°C when determined by a differential scanning calorimetry (DSC) thermogram. In some embodiments, crystalline morphology V is further characterized by a melting point onset of approximately 135.8°C when determined by a differential scanning calorimetry (DSC) thermogram.

[0108] In some embodiments, the crystalline morphology V is further characterized by a differential scanning calorimetry (DSC) thermogram substantially as shown in Figure 9.

[0109] In some embodiments, the crystalline morphology V is characterized substantially by the XRPD pattern shown in Figure 8 and further characterized substantially by the differential scanning calorimetry (DSC) thermogram shown in Figure 9.

[0110] Crystal form V 1 The content of one or more residual solvents (e.g., ethanol and / or ethyl acetate) can be determined using the 1H NMR spectrum. In some embodiments, the crystalline form V is 1 As determined by the 1H NMR spectrum, it has an ethanol content of 7.6%. In some embodiments, crystalline form V is ethyl acetate solvate, ethanol solvate, or a combination thereof. In some embodiments, crystalline form V is ethanol solvate.

[0111] III-5.Crystal form VI In one embodiment, the disclosure provides a crystalline form VI of a compound of formula (I), characterized by an X-ray powder diffraction (XRPD) pattern having three or more peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, 24.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ).

[0112] The crystalline form VI of the compound of formula (I) can be characterized by an X-ray powder diffraction (XRPD) pattern having one or more peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, 24.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ), for example, two, three, four, five, or more peaks, where the XRPD is CuK α1 It is prepared using irradiation. In some embodiments, the crystalline form VI of the compound of formula (I) is characterized by an XRPD pattern containing four or more peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, 24.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form VI of the compound of formula (I) is characterized by an XRPD pattern containing five or more peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, 24.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the crystalline form VI of the compound of formula (I) is characterized by an XRPD pattern containing six or more peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, 24.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ).

[0113] In some embodiments, the crystalline form VI of the compound of formula (I) is characterized by an XRPD pattern containing peaks at 19.4, 21.3, 22.4, and 24.4 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes one or more peaks at 5.8, 10.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes two or more peaks at 5.8, 10.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the XRPD pattern further includes three or more peaks at 5.8, 10.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ).

[0114] In some embodiments, the crystal morphology VI is characterized by an XRPD pattern containing peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, and 24.4 degrees 2θ (±0.2 degrees 2θ).

[0115] In some embodiments, the crystal morphology VI is substantially characterized by the XRPD pattern shown in Figure 10.

[0116] In some embodiments, crystalline form VI substantially does not include other crystalline or amorphous forms of the compound of formula (I).

[0117] In some embodiments, crystalline morphology VI is characterized by a differential scanning calorimetry (DSC) thermogram that includes endothermic activity between approximately 116°C and approximately 170°C. In some embodiments, crystalline morphology VI is further characterized by a differential scanning calorimetry (DSC) thermogram that includes an endothermic peak at approximately 142.9°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram further includes exothermic activity between approximately 187°C and approximately 210°C. In some embodiments, the differential scanning calorimetry (DSC) thermogram is characterized by an exothermic peak at approximately 193.9°C. In some embodiments, crystalline morphology VI is further characterized by a melting point of approximately 142.9°C when determined by a differential scanning calorimetry (DSC) thermogram. In some embodiments, crystalline morphology VI is further characterized by a melting point onset at approximately 116.6°C when determined by a differential scanning calorimetry (DSC) thermogram.

[0118] In some embodiments, the crystalline morphology VI is further characterized by a differential scanning calorimetry (DSC) thermogram substantially as shown in Figure 11.

[0119] In some embodiments, the crystalline morphology VI is characterized substantially by the XRPD pattern shown in Figure 10 and further characterized substantially by the differential scanning calorimetry (DSC) thermogram shown in Figure 11.

[0120] IV. Process for preparing crystal morphology In another aspect, the present disclosure provides a process for preparing the crystalline form of a compound of formula (I). This process is a) Form a first mixture containing the compound of formula (I) and a solvent at a temperature of at least 20°C, b) Adding a poor solvent to the first mixture to form a second mixture, or a) Form a first mixture containing the compound of formula (I) and a solvent at a temperature of at least 20°C, c) Cooling and stirring the first or second mixture to form a precipitate, d) Isolating the precipitate, e) drying the precipitate to obtain the crystalline form of formula (I), The solvent is C 1~4 Alkyl alcohol, di-(C 1~4 The solvent is an alkyl ether, a 5-6 membered cyclic ether, acetic acid, or water, and the poor solvent is C 5~7 Alkane, C 1~4 Alkyl alcohol, di-(C 1~4 Alkyl ethers, 5-6 membered cyclic ethers, di-(C) 1~4 Alkyl ketone, C 1~4 Alkyl-C(O)OC 1~4 It is an alkyl or aromatic hydrocarbon solvent, However, if the solvent and poor solvent are C 1~4 Alkyl alcohol, di-(C 1~4 It must not be an alkyl ether or a 5-6 membered cyclic ether.

[0121] Generally, the morphology of the starting material (i.e., the compound of formula (I)) is not important for the successful recovery of the crystalline material, but the dynamics of initial dissolution may be affected, and a larger proportion of solvent may be required. For example, the desired crystalline form can be obtained using either an amorphous material obtained by freeze-drying or an existing crystalline material. In some cases, it may be beneficial to first isolate the compound of formula (I) as a first crystalline form and use the first crystalline form as a starting material to obtain the desired crystalline form. In special cases, the desired form is synthesized by heating a metastable form under vacuum.

[0122] Furthermore, the sodium content of the starting material can affect the success of crystallization. Generally, samples with a sodium content of 0.5% by weight or more are more difficult to crystallize, but using more solvent can help mitigate this problem.

[0123] Single solvents and two-component solvent mixtures By using either a single solvent or a mixture of two solvents, the desired crystalline form can be produced using several solvents. In the case of a single solvent, the starting material can be dissolved by heating in a solvent that can form a reasonably concentrated solution, and then cooled to precipitate the desired crystalline form. Suitable solvents for use alone or in mixtures include, but are not limited to, isopropanol, ethanol, methanol, acetonitrile, acetic acid, tetrahydrofuran, and water. Alternatively, the starting material can be dissolved in a solvent that can form a reasonably concentrated solution at ambient temperature, and then the solvent can be slowly evaporated (for example, under ambient conditions, under a flow of an inert gas (i.e., N2 or Ar), or under reduced pressure) to precipitate the desired crystalline form.

[0124] In the case of a two-component solvent mixture, the material is first dissolved in a solvent capable of forming a reasonably concentrated solution, as outlined above, and then a low-polarity solvent (e.g., a poor solvent) in which the material does not readily dissolve is added to precipitate the desired material. If the solvent is initially heated to dissolve the material, the poor solvent can be added while the solution is hot or after it has cooled to, for example, room temperature. In a selected example, the material is dissolved in ethanol at room temperature, and ethyl acetate is added to precipitate the desired crystalline form. Suitable precipitation solvents (as poor solvents) include, but are not limited to, toluene, ethyl acetate, diethyl ether, acetone, methyl tert-butyl ether, isopropanol, pentane, hexane, heptane, and acetonitrile.

[0125] In some embodiments, the solvent is C 1~4 Alkyl alcohol, di-(C 1~4 The solvent is an alkyl ether, or a 5-6 membered cyclic ether, or a mixture thereof. In some embodiments, the solvent is C 1~4 It is an alkyl alcohol or a 5-6 membered cyclic ether. Preferred C 1~4Examples of alkyl alcohols include methanol, ethanol, or iso-propanol, but are not limited to these. 1~4 Examples of alkyl ethers include, but are not limited to, diethyl ether, methyl ethyl ether, or methyl tert-butyl ether. Preferred 5-6 membered cyclic ethers include, but are not limited to, tetrahydrofuran, methyltetrahydrofuran, or dioxane. In some embodiments, the solvent is ethanol, tetrahydrofuran, or a mixture thereof. In some embodiments, the solvent is ethanol. In some embodiments, the solvent is tetrahydrofuran. In some embodiments, the solvent further includes water.

[0126] In some embodiments, the present disclosure relates to a process for preparing crystalline forms II, V, or VI of a compound of formula (I), wherein the process is: a) To form a first mixture containing the compound of formula (I) and ethanol at a temperature of at least 20°C, c) Cooling and stirring the first mixture to form a precipitate, d) Isolating the precipitate, The present invention provides a process comprising: e) drying the precipitate to obtain crystalline form II or V of formula (I).

[0127] The temperature at which step a) is performed may affect the identity of the resulting crystal morphology. While we do not wish to be bound by theory, lower temperatures are preferable for the formation of crystal morphology VI, moderate temperatures are preferable for the formation of crystal morphology V, and higher temperatures are preferable for the formation of crystal morphology II. In some embodiments, step a) is performed at a temperature of 20°C to 30°C. In some embodiments, step a) is performed at a temperature of 30°C to 40°C. In some embodiments, step a) is performed at a temperature of 40°C to 65°C.

[0128] In some embodiments, the present disclosure is a process for preparing crystalline forms I, II, III, IV, or V of a compound of formula (I), the process being: a) Form a first mixture containing the compound of formula (I) and a solvent at a temperature of at least 20°C, b) Adding a poor solvent to the first mixture to form a second mixture, c) Cool the second mixture and stir to form a precipitate, d) Isolating the precipitate, e) Drying the precipitate to obtain crystalline forms I, II, III, IV, or V of formula (I), respectively, The present invention provides a process in which the solvent is ethanol or tetrahydrofuran, and the poor solvent is ethyl acetate, methyl tert-butyl ether, heptane, or toluene.

[0129] In some embodiments, the present disclosure is a process for preparing crystalline form I of a compound of formula (I), the process is a) To form a first mixture containing the compound of formula (I) and ethanol at a temperature of at least 20°C, b) Adding a poor solvent to the first mixture to form a second mixture, c) Cool the second mixture and stir to form a precipitate, d) Isolating the precipitate, e) drying the precipitate to obtain crystalline form I of formula (I), The present invention provides a process in which the poor solvent is ethyl acetate or toluene.

[0130] In some embodiments, the present disclosure is a process for preparing crystalline form I of a compound of formula (I), the process is a) To form a first mixture containing the compound of formula (I) and ethanol at a temperature of at least 20°C, b) Adding ethyl acetate to the first mixture to form a second mixture, c) Cool the second mixture and stir to form a precipitate, d) Isolating the precipitate, e) A process is provided which includes drying the precipitate to obtain crystalline form I of formula (I).

[0131] In some embodiments, the present disclosure is a process for preparing crystalline form I of a compound of formula (I), the process is a) To form a first mixture containing the compound of formula (I) and ethanol at a temperature of at least 20°C, b) Adding toluene to the first mixture to form a second mixture, c) Cool the second mixture and stir to form a precipitate, d) Isolating the precipitate, e) A process is provided which includes drying the precipitate to obtain crystalline form I of formula (I).

[0132] In some embodiments, the present disclosure relates to a process for preparing crystalline forms II, V, or mixtures of forms II and V of a compound of formula (I), the process being: a) To form a first mixture containing the compound of formula (I) and ethanol at a temperature of at least 20°C, b) Adding heptane to the first mixture to form a second mixture, c) Cool the second mixture and stir to form a precipitate, d) Isolating the precipitate, e) A process is provided which includes drying the precipitate to obtain crystalline form II of formula (I).

[0133] In some embodiments, the present disclosure is a process for preparing crystalline form III of a compound of formula (I), the process is a) To form a first mixture containing the compound of formula (I) and ethanol at a temperature of at least 20°C, b) Adding methyl tert-butyl ether to the first mixture to form a second mixture, c) Cool the second mixture and stir to form a precipitate, d) Isolating the precipitate, e) A process is provided which includes drying the precipitate to obtain crystalline form III of formula (I).

[0134] In some embodiments, the present disclosure is a process for preparing crystalline form IV of a compound of formula (I), the process is: a) To form a first mixture containing the compound of formula (I) and tetrahydrofuran at a temperature of at least 20°C, b) Adding ethyl acetate to the first mixture to form a second mixture, c) Cool the second mixture and stir to form a precipitate, d) Isolating the precipitate, e) A process is provided which includes drying the precipitate to obtain crystalline form IV of formula (I).

[0135] In some embodiments, the first mixture comprises the crude compound of formula (I). In other embodiments, the first mixture comprises the crystalline form of the compound of formula (I), for example, the crystalline form of the compound of formula (I) according to this disclosure, or the crystalline form described in International Publication No. 2020 / 123772 (i.e., form A or form B). In some embodiments, the first mixture comprises crystalline forms A, B, I, II, III, IV, V, or VI. In some embodiments, the first mixture comprises crystalline form A or B. In some embodiments, the first mixture comprises crystalline form A. In some embodiments, the first mixture comprises crystalline form B.

[0136] In some embodiments, the first and / or second mixture is a solution containing the compound of formula (I). In some embodiments, the first and / or second mixture is a solution containing the compound of formula (I) and ethanol. In some embodiments, the first and / or second mixture is a turbid solution containing the compound of formula (I) and ethanol. In some embodiments, the first and / or second mixture is a turbid solution containing the compound of formula (I) and tetrahydrofuran.

[0137] Slow evaporation of a saturated solution of the material in a suitable solvent or mixture is also effective in obtaining crystalline materials. Generally, the sample has a low degree of crystallinity when measured by XRPD. Suitable solvents include, but are not limited to, acetone, tetrahydrofuran, ethanol, methanol, acetonitrile, and water.

[0138] Solvent / Poor Solvent Ratio In the case of a two-component solvent mixture, the formation of crystalline morphology may be sensitive to the ratio of solvent to precipitation solvent. For example, when a material is dissolved in ethanol and ethyl acetate is added as the precipitation solvent, the final ethanol-to-ethyl acetate ratio can vary from 4:1 to 1:4, e.g., 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, or 1:4. In some embodiments, the final ethanol-to-ethyl acetate ratio is 2:1. While we do not wish to be bound by theory, solvent systems characterized by an ethanol content higher than the ethyl acetate content tend to be favorable for the formation of crystalline morphology II. In contrast, solvent systems characterized by an ethanol content lower than the ethyl acetate content tend to be favorable for the formation of crystalline morphology I. In one embodiment, the ethanol content is higher than the ethyl acetate content. In another embodiment, the ethanol content is lower than the ethyl acetate content. In some embodiments, the ethanol-to-ethyl acetate ratio is 4:1, 3:1, 2:1, or 1:1. In alternative embodiments, the ethanol to ethyl acetate content is 1:1, 1:2, 1:3, or 1:4.

[0139] In some embodiments, the ratio of ethanol to ethyl acetate is 1:1 to 1:4. In some embodiments, the ratio of ethanol to ethyl acetate is 1:1 to 1:2. In some embodiments, the ratio of ethanol to ethyl acetate is about 5:8. In some embodiments, the ratio of ethanol to toluene is 1:1 to 1:4. In some embodiments, the ratio of ethanol to toluene is about 1:2. In some embodiments, the ratio of ethanol to methyl tert-butyl ether is 1:1 to 1:4. In some embodiments, the ratio of ethanol to methyl tert-butyl ether is about 1:2. In some embodiments, the ratio of ethanol to methyl tert-butyl ether is about 5:14. In some embodiments, the ratio of tetrahydrofuran to ethyl acetate is 1.5:1 to 1:2. In some embodiments, the ratio of tetrahydrofuran to ethyl acetate is about 5:4. In some embodiments, the ratio of ethanol to heptane is 1:1 to 1:4.

[0140] When obtaining crystalline materials using slow evaporation, a mixture of solvents may be used. In some embodiments, the solvent ratio can vary from 4:1 to 1:1. In another embodiment, crystalline materials can be obtained using the slow evaporation of a single solvent.

[0141] Solvent / compound ratio The ratio or concentration of the compound to the solvent may vary depending on the solvent or solvent mixture used. Typical concentrations can range from 250 mg / mL to 10 mg / mL, with higher concentrations being limited by the solubility of the material or the ease of recovering the material after crystallization. For example, approximately 200 mg of amorphous material can be dissolved in 1 mL of ethanol, followed by the addition of a poor solvent (e.g., ethyl acetate, heptane, or toluene) to obtain a crystalline form.

[0142] Temperature control Generally, the maximum heating temperature used in the above method can range from 20°C to the reflux temperature of the solvent. The most typical temperature range is 20°C to 60°C. After the solution is obtained, a precipitation solvent is added as needed, and the mixture is cooled to room temperature. The cooling rate can affect the size, shape, and quality of the crystals. Decomposition may occur if the solution is heated for a long time above 60°C or if it contains a reactive solvent.

[0143] In some embodiments, step a) is performed at a temperature of 20°C to 100°C. In some embodiments, step a) is performed at a temperature of 40°C to 80°C. In some embodiments, step a) is performed at a temperature of 55°C to 60°C. In some embodiments, step a) is performed at a temperature of 35°C to 55°C. In some embodiments, step a) is performed at a temperature of 35°C to 40°C. In some embodiments, step a) is performed at a temperature of 20°C to 25°C.

[0144] In some embodiments, if step b) is present, step b) is performed at a temperature of 20°C to 100°C. In some embodiments, if step b) is present, step b) is performed at a temperature of 40°C to 80°C. In some embodiments, if step b) is present, step b) is performed at a temperature of 55°C to 60°C. In some embodiments, if step b) is present, step b) is performed at a temperature of 35°C to 55°C. In some embodiments, if step b) is present, step b) is performed at a temperature of 35°C to 40°C. In some embodiments, if step b) is present, step b) is performed at a temperature of 20°C to 25°C.

[0145] In some embodiments, steps a) and b) are performed at temperatures of approximately 55°C to approximately 60°C, respectively. In some embodiments, steps a) and b) are performed at temperatures of approximately 35°C to 55°C, respectively. In some embodiments, steps a) and b) are performed at temperatures of approximately 35°C to 40°C, respectively. In some embodiments, steps a) and b) are performed at temperatures of approximately 20°C to 25°C, respectively. Crystallization rate

[0146] Several factors significantly affect the crystallization rate. These include, but are not limited to, the rate of addition of the precipitation solvent, the rate of cooling of the mixture, and the presence of nucleation sites such as dust, seed crystals, or defects on the glass surface. Variations in these parameters can affect the size, shape, and quality of the crystals.

[0147] In some embodiments, step c) is performed by c-1) cooling the first or second mixture to room temperature over a period of 30 minutes to 3 hours, and c-2) stirring at room temperature for a period of 12 to 72 hours to form a precipitate. In some embodiments, step c) is performed by c-1) cooling the first or second mixture to room temperature over a period of 1 to 2 hours, and c-2) stirring at room temperature for a period of about 18 hours to form a precipitate.

[0148] In some embodiments, crystalline seeds of the compound of formula (I) are added during step c).

[0149] Isolation of crystalline form Several methods can be used to isolate the desired crystalline form from the supernatant, including filtration, decantation, and solvent evaporation. Generally, the crystalline form was obtained by recovering any formed solid by vacuum filtration, followed by removal of any residual solvent by air drying and subsequent exposure to high vacuum.

[0150] In some embodiments, the isolation in step d) is carried out by filtration.

[0151] In some embodiments, the drying in step e) is carried out under vacuum at a temperature of about 55°C to about 60°C. In some embodiments, the drying in step e) is carried out under vacuum at ambient temperature, for example, about 20°C to 25°C.

[0152] Table 1 summarizes the preparation of the crystal morphology of formula (I).

[0153] [Table 1-1]

[0154] [Table 1-2] A The isolation process was not optimized for yield. Some product remained on the flask walls during filtration. B Standard addition: Add the poor solvent to a batch of the compound / solvent of formula (I). C Back addition: Add the compound of formula (I) / solvent to a poor solvent batch. D Standard addition: Add the solvent to a batch of the compound of formula (I) in a poor solvent. E Add everything at once: Add all of the compound / solvent / poor solvent of formula (I) at the start. Attempt to solubilize the compound of formula (I) by adding additional solvent at high temperature. F Form A of the compound of formula (I) is disclosed in international application PCT / US2019 / 065916.

[0155] V. Composition The crystalline form of the compound of formula (I) may be in the form of a composition suitable for administration to a subject, or may be used to prepare such a composition (e.g., further treatment with one or more excipients). Generally, such a composition is a “pharmaceutical composition” comprising the compound of formula (I) and one or more pharmaceutically acceptable or physiologically acceptable diluents, carriers, or excipients. In certain embodiments, the compound of formula (I) is present in a therapeutically acceptable amount. The pharmaceutical composition may be used in the methods of the present disclosure, and therefore, for example, the pharmaceutical composition may be administered to a subject ex vivo or in vivo to carry out the therapeutic and prophylactic methods and uses described herein.

[0156] The pharmaceutical compositions of this disclosure can be formulated to suit the intended method or route of administration, and exemplary routes of administration are described herein. Furthermore, the pharmaceutical compositions may be used in combination with other therapeutically active agents or compounds, such as those described herein, to treat or prevent diseases, disorders, and conditions as envisioned by this disclosure.

[0157] A pharmaceutical composition containing an active ingredient (e.g., a compound of formula (I)) may be in a form suitable for oral use, such as tablets, capsules, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups, solutions, microbeads, or elixirs. A pharmaceutical composition intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such a composition may contain one or more agents, such as sweeteners, flavoring agents, coloring agents, and preservatives, in order to provide a pharmaceutically superior and palatable preparation. Tablets, capsules, etc., may contain the active ingredient in combination with non-toxic, pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients may include diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulators and disintegrants, such as corn starch or alginic acid; binders, such as starch, gelatin, or acacia; and lubricants, such as magnesium stearate, stearic acid, or talc.

[0158] Tablets, capsules, etc., suitable for oral administration may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained effect. For example, time-delaying materials such as glyceryl monostearate or glyceryl distearate may be used. They may also be coated by techniques known in the art to form osmotic therapeutic tablets for controlled release. Additional agents include biodegradable or biocompatible particles or polymeric substances to control the delivery of the administered composition, such as polyesters, polyamine acids, hydrogels, polyvinylpyrrolidone, polyanhydrides, polyglycolic acid, ethylene-vinyl acetate, methylcellulose, carboxymethylcellulose, protamine sulfate, or lactide / glycolide copolymers, polylactide / glycolide copolymers, or ethylene vinyl acetate copolymers. For example, oral formulations can be encapsulated in microcapsules prepared using hydroxymethylcellulose or gelatin microcapsules or poly(methylmethchlorate) microcapsules, respectively, by coacervation technology or interfacial polymerization, or in colloidal drug delivery systems. Examples of colloidal dispersion systems include macromolecular complexes, nanocapsules, microspheres, microbeads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. The methods for preparing the above formulations will be apparent to those skilled in the art.

[0159] Formulations for oral use may also be presented as rigid gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, kaolin, or microcrystalline cellulose, or as soft gelatin capsules in which the active ingredient is mixed with a water or oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0160] The aqueous suspension contains the active material in combination with excipients suitable for its preparation. Such excipients may be suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and axia gum; dispersing or wetting agents, such as naturally occurring phosphatides (e.g., lecithin), or condensation products of alkylene oxides and fatty acids (e.g., polyoxyethylene stearate), or condensation products of ethylene oxides and long-chain aliphatic alcohols (e.g., heptadecanoethyleneoxycetanol), or condensation products of ethylene oxides and partial esters derived from fatty acids and hexitol (e.g., sorbitol polyoxyethylene monooleate), or condensation products of ethylene oxides and partial esters derived from fatty acids and hexitol anhydrides (e.g., sorbitan polyethylene monooleate). The aqueous suspension may also contain one or more preservatives.

[0161] Oily suspensions can be formulated by suspending the active ingredient in a vegetable oil, such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil, such as liquid paraffin. Oily suspensions may contain thickeners, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavorings, such as those mentioned above, can be added to provide an oral preparation with a pleasant mouthfeel.

[0162] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water are mixed with a dispersant or wetting agent, a suspending agent, and one or more preservatives to provide an active ingredient. Suitable dispersants or wetting agents and suspending agents are exemplified herein.

[0163] The pharmaceutical compositions of this disclosure may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil, e.g., olive oil or peanut oil, or a mineral oil, e.g., liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, e.g., acacia gum or tragacanth gum; naturally occurring phosphatides, e.g., esters or partial esters derived from soybeans, lecithin, and fatty acids; hexitol anhydrides, e.g., sorbitan monooleate; and condensation products of partial esters with ethylene oxide, e.g., sorbitan polyoxyethylene monooleate.

[0164] A pharmaceutical composition typically comprises a therapeutically effective amount of the CD73 inhibitor intended by this disclosure (i.e., a compound of formula (I)) and one or more pharmaceutically and physiologically acceptable combinations. Suitable pharmaceutically or physiologically acceptable diluents, carriers, or excipients include, but are not limited to, antioxidants (e.g., ascorbic acid and sodium bisulfate), preservatives (e.g., benzyl alcohol, methylparaben, ethyl or n-propyl, p-hydroxybenzoic acid), emulsifiers, suspending agents, dispersants, solvents, fillers, bulking agents, surfactants, buffers, vehicles, diluents, and / or adjuvants. For example, a suitable vehicle may be physiological saline or citrate-buffered saline, which may optionally be supplemented with other materials common in pharmaceutical compositions for parenteral administration. Neutral buffered saline or physiological saline mixed with serum albumin are further exemplary vehicles. Typical buffers that may be used in the compositions of this disclosure include, but are not limited to, pharmaceutically acceptable weak acids, weak bases, or mixtures thereof. For example, the buffer component may be a water-soluble material, such as phosphoric acid, tartaric acid, lactic acid, succinic acid, citric acid, acetic acid, ascorbic acid, aspartic acid, glutamic acid, and salts thereof. Examples of acceptable buffers include Tris buffer, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), 2-(N-morpholino)ethanesulfonate sodium salt (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), and N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS).

[0165] After a pharmaceutical composition has been formulated, it may be stored in a sterile vial as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored in any of the following forms: ready-to-use, lyophilized, requiring reconstitution before use, liquid, or other acceptable forms. In some embodiments, the pharmaceutical composition is provided in single-use containers (e.g., single-use vials, ampoules, syringes, or auto-injectors (e.g., EpiPen®)), while multi-use containers (e.g., multi-use vials) are provided in other embodiments.

[0166] The formulations may also include controlled-release formulations comprising carriers, such as liposomes, hydrogels, prodrugs, and microencapsulation delivery systems, to protect the composition from rapid degradation or elimination from the body. For example, time-delaying materials such as glyceryl monostearate or glyceryl stearate may be used alone or in combination with wax. The crystalline form of the compound of formula (I) can be delivered using a variety of drug delivery devices, including implants (e.g., indwellable pumps) and catheter systems, slow-injection pumps and devices, all of which are well known to those skilled in the art.

[0167] Depot injections, generally administered subcutaneously or intramuscularly, may also be used to release the crystalline form of the compound of formula (I) disclosed herein over a predetermined period of time. Depot injections are typically based on either a solid or an oil and generally contain at least one of the formulation components described herein. Those skilled in the art will be familiar with the possible formulations and the use of depot injections.

[0168] The pharmaceutical composition may be in the form of a sterile aqueous or oily suspension for injection. This suspension may be formulated according to known techniques using suitable dispersants or wetting agents and suspending agents as described herein. The sterile injection preparation may also be a sterile injection solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, such as a solution in 1,3-butane-diol. Acceptable diluents, solvents, and dispersion media that may be used include water, Ringer's solution, isotonic sodium chloride solution, Cremophor EL® (BASF, Parsippany, NJ), or phosphate-buffered saline (PBS). Examples include ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Furthermore, sterile fixative oils are conventionally used as solvents or suspension media. For this purpose, any solvent-free fixative oil containing synthetic monoglycerides or diglycerides can be used. In addition, fatty acids such as oleic acid are used in the preparation of injectable formulations. Long-term absorption of certain injectable formulations can be achieved by including absorption-delaying agents (e.g., aluminum monostearate or gelatin).

[0169] This disclosure intends to administer the crystalline form of the compound of formula (I) in the form of a suppository for rectal administration. The suppository can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at normal temperature but liquid at rectal temperature and therefore melts in the rectum to release the drug. Examples of such materials include, but are not limited to, cocoa butter and polyethylene glycol.

[0170] The crystalline form of the compound of formula (I) contemplated by this disclosure may be in the form of any other suitable pharmaceutical composition currently known or to be developed in the future (e.g., a spray for nasal or inhalation use).

[0171] VI.How to use This disclosure intends to describe the use of the crystalline form of the compounds of formula (I) described herein in the treatment or prevention of a wide range of diseases, disorders, and / or conditions, and / or symptoms thereof. Specific uses are described in detail below, but it should be understood that this disclosure is not limited thereto. Furthermore, a general classification of specific diseases, disorders, and conditions is described below, but some of those diseases, disorders, and conditions may belong to two or more classifications, or not to any of the classifications disclosed.

[0172] The use of the crystalline form of the compound of formula (I) in the therapeutic methods described herein includes the direct use of the crystalline form to a subject (e.g., administration) as described herein, as well as the use of the crystalline form in the preparation of a medicament for the treatment of the indications described herein. In some embodiments, the crystalline form of the compound described herein is stored in the final dosage form to be administered to the subject. In other embodiments, the crystalline form may undergo physical transformation, for example, to an amorphous form, a different crystalline form, or a solubilized form, before administration to the subject.

[0173] Oncology-related disorders. According to this disclosure, the compound of formula (I) (e.g., the crystalline form described herein) is a cancer, for example, of the uterus, cervix, breast, prostate, testes, gastrointestinal tract (e.g., esophagus, cervix, oropharynx, stomach, small or large intestine, colon, or rectum), kidney, renal cells, bladder, bone, bone marrow, skin, head and neck, liver, gallbladder, heart, lung, pancreas, salivary glands, adrenal glands, thyroid, brain (e.g., glioma), ganglia, central nervous system (CNS) and peripheral nervous system (PNS), as well as cancers of the hematopoietic system and immune system (e.g., spleen or It can be used to treat or prevent proliferative conditions or disorders, including cancer of the thymus. The disclosure also provides methods for treating or preventing other cancer-related diseases, disorders, or conditions, including, for example, immunogenic tumors, non-immunogenic tumors, dormant tumors, virus-induced cancers (e.g., epithelial cell carcinoma, endothelial cell carcinoma, squamous cell carcinoma, and papillomavirus), adenocarcinomas, such as pancreatic adenocarcinoma, lymphoma, carcinoma, melanoma, leukemia, myeloma, sarcoma, teratoma, chemically induced cancer, metastasis, and angiogenesis. This disclosure aims to reduce tolerance to tumor cells or cancer cell antigens, for example, by modulating the activity of regulatory T cells and / or CD8+ T cells (see, e.g., Ramirez-Montagut, et al. (2003) Oncogene 22:3180-87 and Sawaya, et al. (2003) New Engl. J. Med. 349:1501-09). In certain embodiments, tumors or cancers are colon cancer, ovarian cancer, breast cancer, melanoma, lung cancer, glioblastoma, and leukemia. The use of the terms cancer-related diseases, disorders, and conditions means a broad range of conditions directly or indirectly related to cancer, including, for example, angiogenesis and precancerous conditions, e.g., dysplasia.

[0174] In certain embodiments, cancer may be metastatic or at risk of metastasis, or may occur in spreading tissues, including blood or bone marrow cancers (e.g., leukemia). In some further embodiments, T cell tolerance can be overcome using the crystalline form of the compound of formula (I).

[0175] In one embodiment, the cancer is a malignant tumor of the gastrointestinal tract, such as pancreatic cancer. In another embodiment, the cancer is metastatic pancreatic adenocarcinoma. In one embodiment, the patient is treated for pancreatic cancer with a compound of formula (I) and an anti-PD-1 antibody. In yet another embodiment, the patient is treated for first-line metastatic pancreatic cancer with a compound of formula (I) and an anti-PD-1 antibody, as well as standard treatments for pancreatic cancer, such as those described herein. The patient may be previously treated or may be treatment-inexperienced.

[0176] In some embodiments, the Disclosure provides methods for treating proliferative conditions, cancer, tumors, or precancerous conditions using a compound of formula (I) (e.g., in the crystalline form described herein) and at least one additional therapeutic or diagnostic agent, examples of which are described elsewhere herein.

[0177] In some embodiments, the methods described herein may be presented as first-line, second-line, or third-line treatments.

[0178] Immune-related disorders and disorders having inflammatory components. As used herein, terms such as “immune disease,” “immune condition,” “immune disorder,” “inflammatory disease,” “inflammatory condition,” and “inflammatory disorder” broadly encompass any immune-related condition (e.g., autoimmune disease) or disorder having inflammatory components that can be treated with a compound of formula (I) (e.g., in the crystalline form described herein) to obtain some therapeutic benefit. Such conditions are often closely intertwined with other diseases, disorders, and conditions. For example, “immune condition” may refer to proliferative conditions, such as cancer, tumors, and angiogenesis, which include infections (acute and chronic), tumors, and cancer that are resistant to eradication by the immune system.

[0179] Compounds of formula (I) (e.g., the crystalline forms described herein) may be used to increase or enhance the immune response, to improve immunization, including increasing vaccine efficacy, and to increase inflammation. Immunodeficiency diseases, immunosuppressive medical treatments, acute and / or chronic infections, and aging-related immunodeficiency may be treated using the compounds disclosed herein. Compounds of formula (I) (e.g., the crystalline forms described herein) may also be used to stimulate the immune system of patients suffering from iatrogenically induced immunosuppression, including those who have undergone bone marrow transplantation, chemotherapy, or radiotherapy.

[0180] In certain embodiments of this disclosure, the compound of formula (I) (e.g., in the crystalline form described herein) is used to increase or enhance the immune response to an antigen by providing adjuvant activity. In certain embodiments, at least one antigen or vaccine is administered to a subject in combination with the compound of formula (I) (e.g., in the crystalline form described herein) to prolong the immune response to the antigen or vaccine. Also provided are therapeutic compositions comprising at least one antigenic agent or vaccine component, including but not limited to viruses, bacteria, and fungi or parts thereof, proteins, peptides, tumor-specific antigens, and nucleic acid vaccines, in combination with the compound of formula (I) (e.g., in the crystalline form described herein).

[0181] Microbial disorders. By inhibiting the immunosuppressive and anti-inflammatory activity of CD73, the Disclosure intends to use compounds of formula (I) (e.g., the crystalline form described herein) in the treatment and / or prevention of any viral, bacterial, fungal, parasitic, or other infectious disease, disorder, or condition in which treatment with a CD73 inhibitor may be beneficial. Examples of such diseases and disorders include HIV and AIDS, staphylococcal and streptococcal infections (Staphylococcus aureus and Streptococcus sanginis, respectively), leishmania, toxoplasmosis, trichomoniasis, giardia, candida albicans, bacillus anthracis, and pseudomonas erginosa. The compounds of the Disclosure may be used to treat sepsis, to reduce or inhibit bacterial growth, and to reduce or inhibit inflammatory cytokines.

[0182] CNS-related and neurological disorders. CD73 inhibition may also be an important therapeutic strategy for patients with neurological, neuropsychiatric, neurodegenerative, or other diseases, disorders, and conditions that have some connection to the central nervous system, including disorders associated with cognitive and motor function impairments. Examples include Parkinson's disease, extrapyramidal syndrome (EPS), dystonia, akathisia, tardive dyskinesia, restless leg syndrome (RLS), epilepsy, periodic limb movement in sleep (PLMS), attention deficit disorder, depression, anxiety disorders, dementia, Alzheimer's disease, Huntington's disease, multiple sarcomas, cerebral ischemia, hemorrhagic stroke, subarachnoid hemorrhage, and traumatic brain injury.

[0183] Other Disorders. Embodiments of this disclosure intend to administer compounds of formula (I) (e.g., the crystalline forms described herein) to subjects for the treatment or prevention of any other disorders which can be benefited by at least some level of CD73 inhibition. Such diseases, disorders, and conditions include, for example, cardiovascular (e.g., cardiac ischemia), gastrointestinal (e.g., Crohn's disease), metabolic (e.g., diabetes mellitus), hepatic (e.g., hepatic fibrosis, NASH, and NAFLD), pulmonary (e.g., COPD and asthma), ophthalmic (e.g., diabetic retinopathy), and renal (e.g., renal failure) disorders.

[0184] In some embodiments, the compound of formula (I) (e.g., the crystalline form described herein) may be used to inhibit statin-induced adenosine production or to reduce or decrease the statin-induced increase in blood glucose in subjects taking statins (e.g., lovastatin and pravastatin).

[0185] Patient selection. In some cases, the methods described herein may be applied to specific patients based, for example, on a biomarker such as CD73, high microsatellite instability, or high tumor mutational load. In some cases, subjects are identified as having tumor gene-driven or tumor gene-dependent cancers with mutations in at least one gene associated with CD73. Test methods for determining CD73 levels and the presence of CD73-related tumor genes are disclosed in International Publications 2020 / 185859 and 2020 / 205527.

[0186] Route of administration This disclosure intends to provide for the administration of crystalline forms of compounds of formula (I) and compositions thereof in any suitable form. In one or more embodiments, the crystalline forms of compounds of formula (I) are useful in the manufacture of pharmaceuticals suitable for administration to a subject. In some embodiments, the crystalline forms of compounds of formula (I) are preserved in the pharmaceutical to be administered to the subject. In other embodiments, the crystalline forms of compounds of formula (I) undergo physical changes, for example, to an amorphous form or a different crystalline form, during the preparation of the pharmaceutical. Preferred routes of administration include oral, parenteral (e.g., intramuscular, intravenous, subcutaneous (e.g., injection or implant), intraperitoneal, intrasternal, intraarticular, intraperitoneal, intracerebral (intraparum) and lateral ventricle), nasal, vaginal, sublingual, intraocular, rectal, topical (e.g., transdermal), intrabuccal, and inhalation. Depot injections, generally administered subcutaneously or intramuscularly, can also be used to release the crystalline forms of compounds of formula (I) disclosed herein over a predetermined period of time.

[0187] Certain embodiments of this disclosure are intended for oral administration. Other embodiments of this disclosure are intended for parenteral administration.

[0188] Ecto5'-nucleotidase and its inhibition Human CD73 (ecto-5'-nucleotidase, also known as NT5E or 5NT) is a 574-amino acid protein (accession number AAH6593). Eukaryotic CD73 functions as a non-covalent homodimer with two structural domains, the N-terminal and C-terminal domains connected by a hinge region that allows the enzyme to undergo large domain transitions and switch its conformation between open and closed states (Knapp, K. et al. (2012) Structure 20:2161-73).

[0189] CD73 inhibitors can modulate purinergic signaling, a type of extracellular signaling mediated by purine nucleotides and nucleosides, such as ATP and adenosine. Purinergic signaling involves the activation of purinergic receptors in and / or neighboring cells, leading to the regulation of cellular function. The enzymatic activity of CD73 plays a strategic role in calibrating the duration, magnitude, and chemical properties of purinergic signals delivered to various cells (e.g., immune cells). Modifications in these enzymatic activities may alter the course or prognosis of several pathophysiological events, including cancer, autoimmune and inflammatory diseases, infections, atherosclerosis, and ischemia-reperfusion injury, suggesting that these ectoenzymes represent novel therapeutic targets for managing various disorders.

[0190] Studies using tissues overexpressing CD73 and CD73 knockout mice have provided evidence that CD73 inhibitors have potential benefits for melanoma, lung cancer, prostate cancer, and breast cancer (e.g., Sadej R. (2006) Melanoma). (See Res 16:213-22). High levels of CD73 expression are associated with tumor neoangiogenesis, invasiveness, resistance to chemotherapy, and metastasis; therefore, CD73 inhibitors can be used to control tumor progression and metastasis. Other potential uses are discussed elsewhere in this specification.

[0191] Although the compound of formula (I) is thought to exert its activity by inhibiting CD73, a precise understanding of the compound's fundamental mechanism of action is not necessary to carry out this disclosure. For example, the compound can also exert its activity, at least partially, through the regulation (e.g., inhibition) of other components of the purinergic signaling pathway (e.g., CD39). The purinergic signaling pathway consists of transporters, enzymes, and receptors responsible (primarily) for the synthesis, release, action, and extracellular inactivation of ATP and its extracellular degradation product, adenosine (Sperlagh, B. et al. (Dec 2012) Neuropsychopharmacologia Hungarica 14(4):231-38). Several potential opportunities exist for the regulation of signaling processes. However, some of these opportunities are more manageable than others.

[0192] VII. Combination Therapy This disclosure envisions the use of compounds of formula (I) (e.g., in the crystalline form described herein) in combination with one or more active therapeutic agents. The additional active therapeutic agents may be small chemical molecules; macromolecules, e.g., proteins, antibodies, peptide bodies, peptides, DNA, RNA, or fragments of such macromolecules; or cell therapies or gene therapies. In such combination therapies, the various active agents often have different complementary mechanisms of action. Such combination therapies may be advantageous by allowing a reduction in the dose of one or more of the agents, thereby reducing or eliminating adverse effects associated with one or more of the agents. The compounds of formula (I) of this disclosure may also be useful in overcoming adenosine-dependent immunosuppression, leading to enhanced therapeutic efficacy of other agents. Furthermore, such combination therapies may have synergistic therapeutic or prophylactic effects on the underlying disease, disorder, or condition.

[0193] As used herein, “combination” means treatments that can be administered separately, for example, treatments that can be formulated separately for separate administration (for example, as may be provided in a kit), and treatments that can be administered together in a single formulation (i.e., a “co-formulation”).

[0194] In certain embodiments, the compounds of formula (I) (e.g., the crystalline forms described herein) are administered or applied sequentially, for example, in which case one agent is administered before one or more other agents. In other embodiments, the agents are administered simultaneously, for example, in which case two or more agents are administered simultaneously or nearly simultaneously, and the two or more agents may exist in two or more separate formulations or be combined into a single formulation (i.e., a co-formulation). Regardless of whether the two or more agents are administered sequentially or simultaneously, they are considered to be administered in combination for the purposes of this disclosure.

[0195] The compound of formula (I) (e.g., the crystalline form described herein) may be used in combination with at least one other (active) agent in any manner appropriate to the circumstances. In one embodiment, treatment with at least one activator and the crystalline form of the compound of formula (I) is maintained over a period of time. In another embodiment, treatment with at least one activator is reduced or discontinued (e.g., if the subject is stable), while treatment with the crystalline form of the compound of formula (I) is maintained in a constant dosing regimen. In yet another embodiment, treatment with at least one activator is reduced or discontinued (e.g., if the subject is stable), while treatment with the crystalline form of the compound of formula (I) is reduced (e.g., lower dose, less frequent dosing, or shorter treatment regimen). In yet another embodiment, treatment with at least one activator is reduced or discontinued (e.g., if the subject is stable), while treatment with the crystalline form of the compound of formula (I) is increased (e.g., higher dose, more frequent dosing, or longer treatment regimen). In yet another embodiment, treatment with at least one activator is maintained, and treatment with the crystalline form of the compound of formula (I) is reduced or discontinued (e.g., lower dose, less frequent dosing, or shorter treatment regimen). In yet another embodiment, treatment with at least one activator and treatment with the crystalline form of the compound of formula (I) are reduced or discontinued (e.g., lower dose, less frequent dosing, or shorter treatment regimen).

[0196] Oncology-related disorders. The disclosure provides a method for treating and / or preventing a proliferative state, cancer, tumor, or precancerous disease, disorder, or condition using a compound of formula (I) (e.g., the crystalline form according to the disclosure) and at least one additional therapeutic or diagnostic agent.

[0197] In some embodiments, one or more of the additional therapeutic agents are immunomodulators. Preferred immunomodulators that may be used in this disclosure include CD40L, B7, and B7RP1; activated monoclonal antibodies (mAbs) against stimulant receptors, e.g., anti-CD40, anti-CD38, anti-ICOS, and anti-4-IBB ligands; dendritic cell antigen loading (in vitro or in vivo); anti-cancer vaccines, e.g., dendritic cell carcinoma vaccines; cytokines / chemokines, e.g., IL1, IL2, IL12, IL18, ELC / CCL19, SLC / CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFNa / b, M-CSF, IL-3, GM-CSF, IL-13, and anti-IL-10; bacterial lipopolysaccharides (LPS); indoleamine 2,3-dioxygenase 1 (IDO1) inhibitors and immunostimulant oligonucleotides.

[0198] In certain embodiments, the present disclosure relates to a method for tumor suppression of tumor growth, wherein a compound of formula (I) (e.g., as described herein) is used in combination with a signal transduction inhibitor (STI) to achieve additive or synergistic suppression of tumor growth. The present invention provides a method comprising administering (as a crystalline form of) a drug. As used herein, the term “signaling inhibitor” refers to a drug that selectively inhibits one or more steps in a signaling pathway. The signal transduction inhibitors (STIs) intended by this disclosure include: (i) BCR-ABL kinase inhibitors (e.g., GLEEVEC®), (ii) Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs) including small molecule inhibitors (e.g., gefitinib, erlotinib, afatinib, and osimertinib) and anti-EGFR antibodies, (iii) Inhibitors of the human epidermal growth factor (HER) family of transmembrane tyrosine kinases, e.g., HER-2 / neu receptor inhibitors (e.g., HERCEPTIN®) and HER-3 receptor inhibitors, (iv) Vascular endothelial growth factor receptor (VEGFR) inhibitors including small molecule inhibitors (e.g., axitinib, sunitinib, and sorafenib) and anti-VEGF antibodies (e.g., bevacizumab), and (v) AKT family kinases or AK (vi) Inhibitors of the T pathway (e.g., rapamycin), (vii) Inhibitors of serine / threonine protein kinase B-Raf (BRAF), e.g., vemurafenib, dabrafenib, and encorafenib, (vii) Inhibitors of rearrangement during transfection (RET), e.g., serpacatinib and pralcetonib, (viii) Inhibitors of tyrosine protein kinase Met (MET) (e.g., tepotinib, tivantinib, cabozantinib, and crizotinib), (ix) Anaplastic lymphoma kinase (x) inhibitors of kinase (ALK) (e.g., ensartinib, ceritinib, loratinib, crizotinib, and brigatinib), (x) inhibitors of the RAS signaling pathway as described elsewhere herein (e.g., inhibitors of KRAS, HRAS, RAF, MEK, and ERK), (xi) FLT-3 inhibitors (e.g., gilteritinib), (xii) inhibitors of Trop-2, e.g., antibody-drug conjugates such as satituzumab govitecan-hziy, (xiii) inhibitors of the JAK / STAT pathway, e.g., JAK inhibitors including tofacitinib and ruxolitinib, or STAT inhibitors such as napabucasin, (xiv) inhibitors of NF-κB, (xv) cell cycle kinase inhibitors (e.g., flavopyridol), (xvi) phosphatidylinositol kinase Examples include kinase (PI3K) inhibitors and (xix) protein kinase B (AKT) inhibitors (e.g., capivacertib, mirancertib). Immunomodulatory agents may also be used in combination with the crystalline forms described herein for the suppression of tumor growth in cancer patients. In one or more embodiments, additional therapeutic agents include inhibitors of EGFR, VEGFR, HER-2, HER-3, BRAF, RET, MET, ALK, RAS (e.g., KRAS, MEK, ERK), FLT-3, JAK, STAT, NF-κB, PI3K, AKT, or any combination thereof.

[0199] In other embodiments, the Disclosure provides a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a compound of formula (I) (e.g., in the crystalline form described herein) and at least one chemotherapeutic agent. Examples of chemotherapeutic agents include alkylating agents, e.g., thiotepa and cyclophosphamide; alkyl sulfonates, e.g., busulfan, improsulfan, and pigosulfan; aziridines, e.g., benzodopa, carbocon, metredopa, and uredopa; ethyleneimines and methylameramines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimemiloromelamamine; and nitrogen mass. Antibiotics, e.g., chlorambucil, chlornafadin, chlorophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobenbitin, fenestrine, prednimustine, trophosphamide, uracil mustard; nitrosourea, e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics, e.g., acrasinomycin, actinomycin, Ausramycin, Azaserin, Bleomycin, Kactinomycin, Calicheamycin, Carabicin, Caminomycin, Cardinophilin, Chromomycin, Dactinomycin, Daunorubicin, Detrevicin, 6-Diazo-5-Oxo-L-Norleucine, Doxorubicin, Epirubicin, Esolubicin, Idarubicin, Marcelomycin, Mitomycin, Mycophenolic acid, Nogaramycin, Olibomycin, Peplomycin, Pofilomycin, Pew Romycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zolubicin; antimetabolites, e.g., methotrexate and 5-fluorouracil (5-FU) with or without leucovorin; folate analogs, e.g., denopterin, methotrexate, pteropterin, trimethrexate; purine analogs, e.g., fludarabine, 6-mercaptopurine, thiamiprine, thioguanine;Pyrimidine analogs, e.g., ancitabine, azacitidine, 6-azauridine, carmoful, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens, e.g., carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; anti-adrenaline, e.g., aminoglutethimide, mitotane, trilostane; folic acid supplements, e.g., floric acid; acegraton; aldofamide glycoside; aminolev Phosphate; Amsacrin; Bestlabsil; Bisanthren; Edatrexate; Defofamine; Demecolsin; Diadiquan; Elformitin; Erliptinium acetate; Etoglucide; Gallium nitrate; Hydroxyurea; Lentinan; Ronidamin; Mitoguazone; Mitoxantrone; Mopidamol; Nitracrine; Pentostatin; Fenamet; Pirarubicin; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; Lazoxane; Schizophyllan; Spirogermanium; Tenuazonic acid; Triadicone ;2,2',2''-Trichlorotriethylamine; Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitractol; Pipobroman; Gacitosine; Arabinoside (Ara-C); Cyclophosphamide; Thiotepa; Taxoids, e.g., Paclitaxel, Nab-Paclitaxel, and Docetaxel; Chlorambucil; Gemcitabine; 6-Thiogunine; Mercaptopurine; Methotrexate; Platinum and platinum-coordinated complexes, e.g., Cisplatin and Carboplatin Binbra Stine; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeroda; ibandronate; CPT11; topoisomerase inhibitors; difluoromethylornithine (DMFO); retinoic acid; esperamycin; capecitabine; anthracycline; and any pharmaceutically acceptable salts, acids, or derivatives of the above, but not limited to these.

[0200] Furthermore, chemotherapeutic agents include anti-hormone agents that act to modulate or inhibit the hormonal effects on tumors, such as anti-estrogen agents including tamoxifen, raloxifene, the aromatase inhibitor 4(5)-imidazole, 4-hydroxytamoxifen, trioxyfen, quioxyfen, onapristone, and toremifene, as well as anti-androgen agents such as abiraterone, apalutamide, darolutamide, flutamide, nilutamide, bicalutamide, leuprolide, enzalutamide, and goserelin, and pharmaceutically acceptable salts, acids, or derivatives of any of the above. In certain embodiments, combination therapy includes the administration of a hormone or related hormone.

[0201] In some embodiments, additional chemotherapeutic agents are selected from nab-paclitaxel, gemcitabine, and combinations thereof. In some embodiments, additional chemotherapeutic agents are selected from enzalutamide, docetaxel, and combinations thereof.

[0202] In some embodiments of the method for treating cancer, administration of a therapeutically effective dose of the compound of formula (I) (e.g., as a crystalline form as described herein) in combination with at least one chemotherapeutic agent results in a greater cancer survival rate than that observed by administering either agent alone. In further embodiments of the method for treating cancer, administration of a therapeutically effective dose of the compound of formula (I) in combination with at least one chemotherapeutic agent results in a greater reduction in tumor size or delay in tumor growth than that observed by administering either agent alone.

[0203] Combinations of the compound of formula (I) (e.g., in the crystalline form described herein) with poly(ADP-ribose) polymerase (PARP) inhibitors are also intended. Exemplary PARP inhibitors intended by this disclosure include olaparib, niraparib, and lucaparib.

[0204] In one or more embodiments, combinations of the compound of formula (I) (e.g., the crystalline form described herein) with inhibitors of Bcl-2 family proteins, such as BCL-2 inhibitors (e.g., venetoclax and navitocrax), and MCL-1 inhibitors are also intended.

[0205] Combinations of the compound of formula (I) (e.g., the crystalline form described herein) with an inhibitor of the CD47-SIRPα pathway (e.g., an anti-CD47 antibody, maglorimab) are also being considered.

[0206] In one or more embodiments, combinations of the compound of formula (I) (e.g., the crystalline form described herein) with a DNA methyltransferase (DNMT) inhibitor or a hypomethylating agent are also considered. Exemplary DNMT inhibitors include decitabine, zebralin, and azacitadine.

[0207] In one or more embodiments, combinations of the compound of formula (I) (e.g., the crystalline form described herein) with a histone deacetylase (HDAC) inhibitor are also intended. Exemplary HDAC inhibitors include vorinostat, gibinostat, avexinostat, panobinostat, bellinostat, and trichostatin A.

[0208] In some embodiments, the compound of formula (I) (e.g., the crystalline form described herein) is combined with a menin-MLL inhibitor.

[0209] In some embodiments, combinations of the compound of formula (I) (e.g., the crystalline form described herein) with an isocitrate dehydrogenase (IDH) inhibitor, such as IDH-1 or IDH-2, are also intended. An exemplary IDH-1 inhibitor is ivosidenib. An exemplary IDH-2 inhibitor is enasidenib.

[0210] Additional therapeutic modalities that may be used in combination with the compound of formula (I) (e.g., the crystalline form described herein) include radiotherapy, surgical excision, monoclonal antibodies against tumor antigens, monoclonal antibody-toxin complexes, T cell adjuvants, bone marrow transplantation, or antigen-presenting cells (e.g., dendritic cell therapy) containing Toll-like receptor (TLR) agonists used to stimulate such antigen-presenting cells. .

[0211] Immune checkpoint inhibitors. This disclosure intends to use the compound of formula (I) (e.g., in the crystalline form described herein) in combination with an immune checkpoint inhibitor.

[0212] Numerous genetic and epigenetic alterations, characteristic of all cancers, provide the immune system with a diverse set of antigens that it can use to distinguish tumor cells from their normal counterparts. In the case of T cells, the final amplitude (e.g., level of cytokine production or proliferation) and quality (e.g., the type of immune response produced, e.g., pattern of cytokine production) of the response initiated through antigen recognition by the T-cell receptor (TCR) are regulated by a balance between co-stimulatory and inhibitory signals (immune checkpoints). Under normal physiological conditions, immune checkpoints are also important for preventing autoimmunity (i.e., maintaining self-tolerance) and protecting tissues from damage when the immune system is responding to pathogenic infection. The expression of immune checkpoint proteins can be dysregulated by tumors as a key immune resistance mechanism.

[0213] Some immune checkpoints (ligands and receptors) are selectively upregulated in various types of tumor cells and are candidates for blockade. Examples include PD-1 (programmed cell death protein 1); PD-L1 (PD-1 ligand); BTLA (B and T lymphocyte attenuation factor); CTLA-4 (cytotoxic T lymphocyte-associated antigen 4); TIGIT (T cell immune receptor with Ig and ITIM domains); TIM-3 (T cell membrane protein 3); LAG-3 (lymphocyte activation gene 3); A2aR (adenosine A2a receptor A2aR); and, based on their structural characteristics, i) killer cell immunoglobulin-like receptors (KIRs), and ii) type C lectin receptors (type II transmembrane receptors). Killer inhibitory receptors can be classified into two classes (members of the receptor family). Other less well-defined immune checkpoints are described in the literature and include both receptors (e.g., the 2B4 receptor (also known as CD244)) and ligands (e.g., certain B7 family inhibitory ligands, e.g., B7-H3 (also known as CD276) and B7-H4 (also known as B7-S1, B7x, and VCTN1)). See Pardoll, (April 2012) Nature Rev. Cancer 12:252-64.

[0214] This disclosure intends to describe the use of the aforementioned immune checkpoint receptor and ligand inhibitors, as well as compounds of formula (I) in combination with immune checkpoint receptors and ligands not yet described (e.g., in the crystalline form described herein). Certain regulators of immune checkpoints are currently available, while others are in late-stage development. For example, when the fully humanized CTLA-4 monoclonal antibody ipilimumab (YERVOY®, Bristol-Myers Squibb) was approved in 2011 for the treatment of melanoma, it became the first immune checkpoint inhibitor to receive regulatory approval in the United States. Fusion proteins containing CTLA-4 and antibodies (CTLA4-Ig, abatocept (ORENCIA®, Bristol-Myers Squibb)) are used to treat rheumatoid arthritis, and other fusion proteins have been shown to be effective in kidney transplant patients susceptible to Epstein-Barr virus. The following classes of immune checkpoint inhibitors approved by the authorities were against PD-1 and its ligands PD-L1 and PD-L2. Approved anti-PD1 antibodies include nivolumab (OPDIVO, Bristol-Myers Squibb) and pembrolizumab (KEYTRUDA®, Merck) for various cancers, including squamous cell carcinoma, classical Hodgkin lymphoma, and urothelial carcinoma. Approved anti-PDL1 antibodies include avelumab (BAVENCIO®, EMD Serono & Examples include Pfizer, atezolizumab (TECENTRIQ®, Roche / Genentech), and durvalumab (IMFINZI®, AstraZeneca), which target certain cancers, including urothelial carcinoma. There are no approved therapies targeting TIGIT or its ligands CD155 and CD112, but those in development include BMS-986207 (Bristol-Myers Squibb), MTIG7192A / RG6058 (Roche / Genentech), OMP-31M32 (OncoMed), and dombanalimab (AB154).In some combinations provided herein, the immune checkpoint inhibitors include ipilimumab, tremelimumab, BMS-986016, IMP-731, IMP-321, covolimab, MBG453, Sym023, INCAGN2390, LY3321367, BMS, 986258, SHR1702, MEDI-0680, and pizilizumab (CT-01 1) Select from nivolumab, pembrolizumab, avelumab, atezolizumab, buzigalimab, BI-75091, chamelerizumab, cosiberimab, durvalumab, dostallimab, semiprimab, cintilimab, tislerizumab, tripalimab, retifanlimab, sasanlimab, dombanarimab (AB154), and zimbererimab (AB122).

[0215] In one aspect of the present disclosure, a compound of formula (I) (e.g., a crystalline form described herein) is combined with an immuno-oncology agent that is either (i) an agonist of a stimulatory (including co-stimulatory) receptor or (ii) an antagonist of an inhibitory (including co-inhibitory) signal to T cells, both of which result in amplification of an antigen-specific T cell response. Certain stimulatory and inhibitory molecules are members of the immunoglobulin super family (IgSF). One important family of membrane-bound ligands that bind to co-stimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), B7-H6, and B7-H7 (HHLA2). Another family of membrane-bound ligands that bind to co-stimulatory or co-inhibitory receptors is the molecules of the TNF family that bind to members of the homologous TNF receptor family, which includes CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR / Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LT13R, LIGHT, DcR3, HVEM, VEGI / TL1A, TRAMP / DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin a / TNF13, TNFR2, TNFa, LT13R, lymphotoxin a 1132, FAS, FASL, RELT, DR6, TROY, NGFR.

[0216] In another aspect, the immuno-oncology agent is a cytokine that inhibits T cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and other immunosuppressive cytokines) or a cytokine that stimulates T cell activation to stimulate an immune response.

[0217] In one aspect, the T cell response can be stimulated by a combination of an oral formulation comprising a compound of formula (I) and a chelating agent, and one or more of (i) a protein that inhibits T cell activation, such as an antagonist of CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, galectin 9, CEACAM-1, BTLA, CD69, galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4 (e.g., an immune checkpoint inhibitor), and / or (ii) a protein that stimulates T cell activation, such as an agonist of B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD2. Other agents that can be combined with the oral formulation comprising the compound of formula (I) and a chelating agent for the treatment of cancer include an antagonist of an inhibitory receptor on NK cells or an agonist of an activating receptor on NK cells. For example, the compounds herein can be combined with an antagonist of KIR, such as lirilumab.

[0218] Still other agents for combination therapy include agents that inhibit or deplete macrophages or monocytes, including CSF-1R antagonist antibodies, including CSF-1R antagonists such as RG7155 (WO 2011 / 70024, WO 2011 / 107553, WO 2011 / 131407, WO 2013 / 87699, WO 2013 / 119716, WO 2013 / 132044) or FPA-008 (WO 2011 / 140249, WO 2013 / 169264, WO 2014 / 036357), but are not limited thereto.

[0219] In another embodiment, the compound of formula (I) (e.g., the crystalline form described herein) may be used in conjunction with one or more agents that ligate positive costimulatory receptors, blockers that attenuate signaling through inhibitory receptors, antagonists, and agents that systemically increase the frequency of antitumor T cells, agents that overcome distinct immunosuppressive pathways within the tumor microenvironment (e.g., blocking inhibitory receptor binding (e.g., PD-L1 / PD-1 interaction), depleting or inhibiting Tregs (e.g., using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by ex vivo anti-CD25 bead depletion), or reversing / preventing T cell anergy or exhaustion), and agents that trigger innate immune activation and / or inflammation at the tumor site.

[0220] Immunomodulators. This disclosure envisions the use of compounds of formula (I) (e.g., in the crystalline form described herein) in combination with therapeutic agents that modulate the tumor microenvironment or enhance or mediate the immune response. Examples of such agents include indoleamine 2,3-dioxygenase 1 (IDO-1) inhibitors, adenosine receptor antagonists, and arginase inhibitors. IDO-1 degrades tryptophan, which impairs the activation of antitumor T cells. Similarly, arginases have been shown to be involved in tumor immune evasion via ARG-1, by depleting arginine from the tumor microenvironment, leading to T cell dysfunction, e.g., cessation of cytokine proliferation and secretion. Exemplary arginase compounds can be found, for example, in International Publication Nos. 2019 / 173188 and 2020 / 102646. 2A R and A 2B Adenosine signaling via R impairs the maturation and / or activation of T cells, NK cells, and dendritic cells, which in turn impairs the activation of the immune system against cancer cells. In some embodiments, this disclosure intends to be used in combination with adenosine receptor antagonists described in International Publication No. / 2018 / 136700, 2018 / 204661, 2018 / 213377, or 2020 / 023846.

[0221] In certain embodiments, the Disclosure envisions the use of a compound of formula (I) (e.g., in the crystalline form described herein) in combination with other agents that modulate adenosine levels. Such therapeutic agents may act on other ectonucleotides that catalyze the conversion of ATP to adenosine, including ectonucleoside triphosphate diphosphohydrolase 1 (ENTPD1, ​​also known as CD39 or differentiation cluster 39), which hydrolyzes ATP to ADP and ADP to AMP.

[0222] In certain embodiments, the present invention intends to use a compound of formula (I) (e.g., the crystalline form described herein) together with an inhibitor of HIF-2α, which plays a crucial role in the cellular response to low oxygen availability. Under hypoxic conditions, hypoxia-inducible factor (HIF) transcription factors are involved in metabolism, angiogenesis, cell proliferation and survival, and immunity. It can activate the expression of genes that regulate avoidance and inflammatory responses. HIF-2α overexpression is associated with poor clinical outcomes in patients with various cancers, and hypoxia is also common in many acute and chronic inflammatory disorders, such as inflammatory bowel disease and rheumatoid arthritis. Exemplary HIF-2α inhibitors include berzutifan, ARC-HIF2, PT-2385, and those described in international applications PCT / US2020 / 063000 and PCT / US2021 / 022912.

[0223] In certain embodiments, this disclosure intends to utilize compounds of formula (I) (e.g., the crystalline form described herein) in combination with inhibitors of phosphatidylinositol 3-kinase (PI3K), particularly PI3Kγ isoforms. PI3Kγ inhibitors can stimulate an anti-cancer immune response by modulating myeloid cells, for example, by inhibiting suppressive myeloid cells and weakening immunosuppressive tumor-infiltrating macrophages, or by stimulating macrophages and dendritic cells to produce cytokines that contribute to an effective T-cell response, resulting in a reduction of cancer development and spread. In one embodiment, the PI3Kγ inhibitor is IPI-549. In another embodiment, the PI3K inhibitor is selected from those described in international application PCT / US2020 / 035920.

[0224] This disclosure also envisions combinations of a compound of formula (I) (e.g., the crystalline form described herein) with one or more RAS signaling inhibitors. Tumor-genic mutations in RAS family genes, e.g., HRAS, KRAS, and NRAS, are associated with various cancers. For example, mutations in KRAS family genes, particularly G12C, G12D, G12V, G12A, G13D, Q61H, G13C, and G12S, have been observed in multiple tumor types. Direct and indirect inhibitory strategies have been investigated for inhibiting mutant RAS signaling. Indirect inhibitors target non-RAS effectors in the RAS signaling pathway, including, but not limited to, inhibitors of RAF, MEK, ERK, PI3K, PTEN, SOS (e.g., SOS1), mTORC1, SHP2 (PTPN11), and AKT. Non-exclusive examples of indirect inhibitors under development include RMC-4630, RMC-5845, RMC-6291, RMC-6236, JAB-3068, JAB-3312, TNO155, RLY-1971, and BI1701963. Direct inhibitors of RAS variants are also being explored, generally targeting the KRAS-GTP or KRAS-GDP complex. Exemplary direct RAS inhibitors under development include, but are not limited to, sotrasib (AMG510), MRTX849, mRNA-5671, and ARS1620. In some embodiments, one or more RAS signaling inhibitors are selected from the group consisting of RAF inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, PTEN inhibitors, SOS1 inhibitors, mTORC1 inhibitors, SHP2 inhibitors, and AKT inhibitors. In other embodiments, one or more RAS signaling inhibitors directly inhibit RAS variants.

[0225] In some embodiments, the present disclosure relates to a compound of formula (I) (e.g., the crystalline form described herein) and one or more inhibitors of anexelekto (i.e., AXL) The combination of these factors is targeted. The AXL signaling pathway is associated with tumor growth and metastasis and is thought to mediate resistance to various cancer therapies. There are various AXL inhibitors under development that also inhibit other kinases within the TAM family (i.e., TYRO3, MERTK), as well as other receptor tyrosine kinases, including MET, FLT3, RON, and AURORA, among others. Exemplary multi-kinase inhibitors include gilteritinib, merestinib, cabozantinib, BMS777607, and foretinib. AXL-specific inhibitors, such as SGI-7079, TP-0903 (i.e., davermatinib), BGB324 (i.e., vemcentinib), and DP3975, are also under development.

[0226] This disclosure also envisions combinations of a compound of formula (I) (e.g., in the crystalline form described herein) with one or more p21-activated kinase 4 (PAK4) inhibitors. PAK4 overexpression has been shown across various cancer types, particularly those resistant to PD-1 therapy. While there are no approved PAK4 inhibitors, several are under development and exhibit dual PAK4 / NAMPT inhibitor activity, e.g., ATG-019 and KPT-9274. In some embodiments, the compounds according to this disclosure are combined with PAK4-selective inhibitors. In some embodiments, the compounds according to this disclosure are combined with PAK4 / NAMPT dual inhibitors, e.g., ATG-019 or KPT-9274.

[0227] Metabolic and cardiovascular diseases. This disclosure provides methods for treating and / or preventing certain cardiovascular and / or metabolic-related diseases, disorders, and conditions, and associated disorders, with a compound of formula (I) (e.g., in the crystalline form described herein) and at least one additional therapeutic or diagnostic agent.

[0228] Examples of therapeutic agents useful in combination therapy for the treatment of hypercholesterolemia (and atherosclerosis) include statins that inhibit the enzymatic synthesis of cholesterol (e.g., CRESTOR®, LESCOL®, LIPITOR®, MEVACOR®, PRAVACOL®, and ZOCOR®); bile acid resins that block cholesterol and prevent its absorption (e.g., COLESTID, LO-CHOLEST, PREVALITE®, QUESTRAN®, and WELCHOL®); ezetimibe (ZETIA®) that blocks cholesterol absorption; fibrinic acid (e.g., TRICOR®) that can reduce triglycerides and moderately increase HDL; niacin (e.g., NIACOR®) that moderately lowers LDL cholesterol and triglycerides; and / or combinations thereof (e.g., VYTORIN® (ezetimibe and simvastatin)). Potential alternative cholesterol treatments for use in combination with the CD73 inhibitors described herein include a variety of supplements and herbs (e.g., garlic, policosanol, and guggul).

[0229] This disclosure includes any pharmaceutically acceptable salts, acids, or derivatives of any of the above.

[0230] Immune-related disorders and disorders having inflammatory components. The disclosure provides methods for treating and / or preventing immune-related diseases, disorders, and conditions, as well as diseases, disorders, and conditions having inflammatory components, with a compound of formula (I) (e.g., in the crystalline form described herein) and at least one additional therapeutic or diagnostic agent.

[0231] Examples of therapeutic agents useful in combination therapy for immune and inflammation-related diseases, disorders, or conditions include, but are not limited to, non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin, ibuprofen, and other propionic acid derivatives (aluminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, indoprofen, ketoprofen, miloprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid, and thioxaprofen), and acetic acid derivatives (indomethacin, acemetacin, alclofenac, clidanac, diclofenac, fenclofenac, fenclodic acid, fentia). Zac, filofenac, ibufenac, isoxepac, oxypinac, sulindac, thiopinac, tolmetine, didomethacin, and zomepirac), fenamic acid derivatives (flufenamic acid, meclofenamic acid, mefenamic acid, diflumic acid, and tolfenamic acid), biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicam (isoxicam, piroxicam, sudoxicam, and tenoxicam), salicylates (acetylsalicylic acid, sulfasalazine), and pyrazoles (apazon, bezpiperilone, feprazon, mofebutazone, oxyfenbutazone, phenylbutazone). Other combinations include cyclooxygenase-2 (COX-2) inhibitors.

[0232] Other activators for combinations include steroids, such as prednisolone, prednisone, methylprednisolone, betamethasone, dexamethasone, or hydrocortisone. Such combinations may be particularly advantageous because one or more adverse effects of the steroids can be reduced or even eliminated by gradually decreasing the required steroid dose.

[0233] For example, an additional example of an active agent that may be used in combination to treat rheumatoid arthritis is a cytokine-suppressive anti-inflammatory drug. Examples include antibodies or antagonists against other human cytokines or growth factors, such as TNF, LT, IL-10, IL-2, IL-6, IL-7, IL-8, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF, or PDGF.

[0234] Certain combinations of activators can interfere with different aspects of the autoimmune and subsequent inflammatory cascade, including TNF antagonists, e.g., chimeric, humanized, or human TNF antibodies, REMICADE®, HUMIRA®, anti-TNF antibody fragments (e.g., CDP870), and soluble p55 or p75 TNF receptors, their derivatives, p75TNFRIgG (ENBREL® or p55TNFR1gG (LENERCEPT), soluble IL-13 receptor (sIL-13), and TNFα-converting enzyme (TACE) inhibitors). Similarly, IL-1 inhibitors (e.g., interleukin-1-converting enzyme inhibitors) may also be effective. Other combinations include interleukin-11, anti-P7s, and p-selectin glycoprotein ligands (PSGLs). Other examples of drugs useful in combination with the crystalline forms described herein include interferon-131a (AVONEX®); interferon-13lb (BETASERON®); copaxone; hyperbaric oxygen; intravenous immunoglobulin; clabribine; and antibodies or antagonists against other human cytokines or growth factors (e.g., antibodies against CD40 ligand and CD80).

[0235] Microbial diseases. This disclosure provides methods for treating and / or preventing viral, bacterial, fungal, and parasitic diseases, disorders, and conditions, and related disorders, with a compound of formula (I) (e.g., in the crystalline form described herein) and at least one additional therapeutic or diagnostic agent (e.g., one or more other antiviral agents and / or one or more agents not associated with viral therapy).

[0236] Such combination therapies include, but are not limited to, antiviral agents that target various stages of the viral survival cycle and have different mechanisms of action, including: inhibitors of viral uncoating (e.g., amantadine and rimantidine); reverse transcriptase inhibitors (e.g., acyclovir, zidovudine, and lamivudine); integrase-targeting agents; agents that block the binding of transcription factors to viral DNA; agents that affect translation (e.g., antisense molecules) (e.g., homivirsen); agents that modulate translation / ribozyme function; protease inhibitors; viral assembly modifiers (e.g., rifampicin); antiretroviral drugs, e.g., nucleoside analog reverse transcriptase inhibitors (e.g., azidothymidine (AZT), ddl, ddC, 3TC, d4T), etc.; non-nucleoside reverse transcriptase inhibitors (e.g., efavirenz, nevirapine); nucleotide analog reverse transcriptase inhibitors; and agents that prevent the release of viral particles (e.g., zanamivir and oseltamivir). The treatment and / or prevention of certain viral infections (e.g., HIV) often involves the use of a group of antiviral drugs ("cocktails").

[0237] Other antiviral agents intended for use in combination with the compound of formula (I) (e.g., the crystalline form described herein) include, but are not limited to, abacavir, adefovir, amantadine, amprenavir, amprigen, arbidol, atazanavir, atripra, boceprevirertet, and cidofovir. , Combivir, darunavir, delavirdine, didanosine, docosanol, edoxudine, emtricitabine, enfuvirtide, entecavir, famciclovir, fosamprenavir, foscarnet, phosphonete, ganciclovir, ibacitabine, immunovir, idoxuridine, imiquimod, indinavir, inosine, various interferons (e.g., peginterferon alfa-2a), lopinavir, lobridine, maraviroc, moroxydine, methylthiazone, nelfinavir, nexavir, penciclovir, peramivir, pre-conazole, podophyllotoxin, raltegravir, ribavirin, ritonavir, pyrimidine, saquinavir, stavudine, telaprevir, tenofovir, tipranavir, trifluridine, triduvir, tromantadine, torvada, valaciclovir, valganciclovir, vicriviroc, vidarabine, viramidine, and zalcitabine.

[0238] ]]The present disclosure contemplates the use of a compound of formula (I) (e.g., the crystalline forms described herein) in combination with an anti-parasitic agent. Such agents include, but are not limited to, thiabendazole, pyrantel pamoate, mebendazole, praziquantel, niclosamide, bithionol, oxamniquine, metrifonate, ivermectin, albendazole, eflornithine, melarsoprol, pentamidine, benznidazole, nifurtimox, and nitroimidazole. One skilled in the art will recognize other agents that may be useful in the treatment of parasitic disorders.

[0239] Embodiments of this disclosure envision the use of a compound of formula (I) (e.g., the crystalline form described herein) in combination with an agent useful in the treatment or prevention of bacterial damage. Antibacterial agents can be classified in various ways, including based on their mechanism of action, chemical structure, and active range. Examples of antibacterial agents include those that target the bacterial cell wall (e.g., cephalosporins and penicillins) or the cell membrane (e.g., polymyxins), or those that block essential bacterial enzymes (e.g., sulfonamides, rifamycin, and quinolines). Most antibacterial agents that target protein synthesis (e.g., tetracyclines and macrolides) are bacteriostatic, while agents such as aminoglycosides are bactericidal. Another means of classifying antibacterial agents is based on their target specificity; “narrow-spectrum” agents target specific types of bacteria (e.g., Gram-positive bacteria, e.g., Streptococcus), while “broad-spectrum” agents have activity against a wider range of bacteria. Those skilled in the art will recognize the types of antibacterial agents suitable for use in specific bacterial infections.

[0240] Embodiments of this disclosure envision the use of compounds of formula (I) (e.g., in the crystalline form described herein) in combination with agents useful in the treatment or prevention of fungal infections. Examples of antifungal agents include polyenes (e.g., amphotericin, nystatin, and pimaricin); azoles (e.g., fluconazole, itraconazole, and ketoconazole); allylamines (e.g., naphthifine and terbinafine) and morpholines (e.g., amorolfine); and antimetabolites (e.g., 5-fluorocytosine).

[0241] Other therapeutic modalities. In another embodiment, the disclosure intends to use the compound of formula (I) (e.g., the crystalline form described herein) in combination with adoptive cell therapy, a novel and promising form of personalized immunotherapy in which immune cells having antitumor activity are administered to cancer patients. Adoptive cell therapy may include, for example, chimeric antigen receptors (CARs) or Therapies using tumor-infiltrating lymphocytes (TILs) and T cells engineered to express T cell receptors (TCRs) are being explored. Adoptive cell therapy generally involves collecting T cells from an individual, genetically modifying them to target specific antigens or enhance their antitumor effects, amplifying them to a sufficient number, and injecting the genetically modified T cells into cancer patients. T cells can be collected from patients who will later reinject the augmented cells (e.g., autologous) or from donor patients (e.g., allogeneic).

[0242] In certain embodiments, this disclosure envisions the use of a compound of formula (I) (e.g., the crystalline form described herein) in combination with an RNA interference-based therapeutic for silencing gene expression. RNAi is initiated by cleaving a long double-stranded RNA into small interfering RNA (siRNA). One strand of the siRNA is connected to a ribonucleoprotein complex known as the RNA-induced silencing complex (RISC). It is incorporated into the fusion and then used to identify mRNA molecules that are at least partially complementary to the incorporated siRNA chain. RISC can either bind to the mRNA or cleave it, either of which inhibits translation.

[0243] This disclosure includes pharmaceutically acceptable salts, acids, or derivatives of the above-mentioned drugs (and members of the class of drugs).

[0244] dosage The compound of formula (I) (e.g., the crystalline form described herein) may be administered to a subject in an amount appropriate to, for example, the target of administration (e.g., the desired degree of degradation); the age, weight, sex, and health and physical condition of the subject to whom the formulation is administered; the route of administration; and the nature of the disease, disorder, condition, or symptoms thereof. The dosage regimen may also take into account the presence, nature, and extent of any adverse effects associated with the administered drug. Effective dosages and dosage regimens can be readily determined, for example, from safety and dose escalation studies, in vivo studies (e.g., animal models), and other methods known to those skilled in the art.

[0245] Generally, drug administration parameters indicate that the dosage is lower than the maximum tolerated dose (MTD), which is the amount that can be irreversibly toxic to the subject, and not lower than the amount required to produce a measurable effect on the subject. Such a amount is determined, for example, by pharmacokinetic and pharmacodynamic parameters related to ADME, taking into account the route of administration and other factors.

[0246] The effective dose (ED) is the amount of the substance taken by a certain percentage of the population. The median effective dose or ED50 of a drug is the dose or amount of a drug that produces a therapeutic response or the desired effect in 50% of the population to which it is administered. While ED50 is commonly used as a reasonable measure of the expected effect of a drug, it is not necessarily the dose that a clinician can deem appropriate after considering all relevant factors. Therefore, in some situations the effective dose may be greater than the calculated ED50, in other situations the effective dose may be less than the calculated ED50, and in yet other situations the effective dose may be the same as the calculated ED50.

[0247] Furthermore, the effective dose of the compound of formula (I) (e.g., the crystalline form described herein) may be an amount that, when administered to a subject in one or more doses, produces the desired result for a healthy subject. For example, for a subject experiencing a particular disorder, the effective dose may be such that the diagnostic parameters, scales, markers, etc. of the disorder are improved by at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, where 100% is defined as the diagnostic parameters, scales, markers, etc. exhibited by a normal subject.

[0248] In certain embodiments, the compound of formula (I) (e.g., the crystalline form described herein) may be administered once or more times per day at a dosage level of about 0.01 mg / kg to about 50 mg / kg, or about 1 mg / kg to about 25 mg / kg, based on the daily body weight of the subject, in order to obtain the desired therapeutic effect (e.g., orally or parenterally).

[0249] For oral administration, the composition may be provided in the form of tablets, capsules, etc., containing 1 to 1000 milligrams of the active ingredient, particularly 1, 3, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, and 1000 milligrams of the active ingredient. In some embodiments, the composition contains 25 to 350 milligrams of the activator. In some embodiments, the composition contains 50 milligrams. In some embodiments, the composition contains 100 milligrams of the activator. In some embodiments, the composition contains 300 milligrams of the activator.

[0250] For parenteral administration of the compound of formula (I), the compound (e.g., in the crystalline form or lyophilized form described herein) may be provided before its reconstitution in a suitable vehicle. In some embodiments, the compound of formula (I) is provided in amounts of 1 to 1000 milligrams of the active ingredient, particularly 1, 3, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, and 1000 milligrams. In some embodiments, the compound of formula (I) is provided in amounts of about 25 to 350 milligrams. In some embodiments, the compound is provided in amounts of 25 to 120 milligrams. In some embodiments, the compound is provided in amounts of 25 to 110 milligrams. In some embodiments, the compound is provided in amounts of 25 to 100 milligrams.

[0251] In some embodiments, the compound of formula (I) (e.g., the crystalline form described herein) may be administered on a monthly, weekly, or daily basis (e.g., orally or parenterally). In some embodiments, the compound of formula (I) may be administered at least once a month, for example, twice a month, three times a month, four times a month, once a week, or daily. In some embodiments, the compound of formula (I) may be administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, or once every six weeks. In certain embodiments, the compound of formula (I) may be administered once or more times a day (e.g., orally). In some embodiments, the compound of formula (I) may be administered once, twice, or three times a day (e.g., orally). In some embodiments, the compound of formula (I) may be administered once a day (e.g., orally). In some embodiments, the compound of formula (I) may be administered once, twice, three times, or four times per month (e.g., parenterally). In some embodiments, the compound of formula (I) may be administered once a week (e.g., parenterally).

[0252] In certain embodiments, an oral formulation containing the compound of formula (I) (e.g., the crystalline form described herein) is administered so that a dose of 50 mg to 350 mg, for example, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, or 350 mg of the crystalline form of the compound of formula (I) is administered daily. In one embodiment, the oral formulation is administered so that a dose of 100 mg of the compound of formula (I) is administered daily. In another embodiment, the oral formulation is administered so that a dose of 300 mg of the compound of formula (I) is administered daily.

[0253] In certain embodiments, the dosage of the compound of formula (I) (e.g., the crystalline form described herein) is contained in a “unit dosage form.” The term “unit dosage form” refers to a physically distinct unit, each unit containing a predetermined amount of the compound of formula (I) (e.g., the crystalline form described herein) alone or in combination with one or more additional agents, sufficient to produce the desired effect. The predetermined amount of the compound of formula (I) in a unit dosage form may be equal to the desired dosage or fraction thereof. For example, a unit dosage form may contain the desired dose, or 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, or 1 / 8 of the desired dose. In certain such embodiments, a unit dosage form may be administered once, twice, three times, four times, five times, six times, seven times, or eight times, respectively, to achieve the desired dose of the active ingredient. In one or more embodiments, the predetermined amount of the compound of formula (I) in a unit dosage form is equal to or half of the desired dose. In certain such embodiments, the unit dosage form may be administered once or twice to achieve the desired dose of the active ingredient. It will be understood that the parameters of the unit dosage form will depend on the specific drug and the action to be achieved.

[0254] VIII. Kit This disclosure also intends to provide kits comprising compounds of formula (I) (e.g., in the crystalline form described herein) and pharmaceutical compositions thereof. Kits generally take the form of physical structures containing various components, as described below, and may be used, for example, when carrying out the methods described above.

[0255] The kit may contain a compound of formula (I) disclosed herein (e.g., provided in a sterile container), which may be in the form of a pharmaceutical composition suitable for administration to a subject. The crystalline form of the compound of formula (I) may be provided in a ready-to-use form (e.g., a tablet or capsule) or in a form that requires reconstitution or dilution before administration (e.g., a powder). If the crystalline form of the compound of formula (I) is in a form that requires reconstitution or dilution by the user, the kit may also include a diluent (e.g., sterile water), buffers, pharmaceutically acceptable excipients, etc., packaged together with or separately from the crystalline form of the compound of formula (I). If combination therapy is intended, the kit may contain several agents separately or they may already be combined within the kit. Each component of the kit may be sealed in an individual container, and all of the various containers may be in a single package. The kits of this disclosure may be designed for conditions necessary to properly maintain the components contained therein (e.g., refrigeration or freezing).

[0256] The kit may include a label or accompanying information sheet containing information identifying its components and instructions for their use (e.g., dosage parameters, clinical pharmacology of the active ingredient, including mechanism of action, pharmacokinetics and pharmacodynamics, adverse effects, contraindications, etc.). The label or accompanying information sheet may include manufacturer information such as lot number and expiration date. The label or accompanying information sheet may be, for example, incorporated into the physical structure containing the components, contained separately within the physical structure, or attached to the components of the kit (e.g., ampoules, tubes, or vials).

[0257] Labels or accompanying documents may further include or be incorporated into computer-readable media. In some embodiments, the actual instructions are not present in the kit, but means are provided for obtaining the instructions from a remote source, for example, via the internet. [Examples]

[0258] IX. Examples The following examples are provided to give a complete disclosure and explanation of the methods of preparation and use of the present disclosure to those skilled in the art, and are not intended to limit the scope of what the inventors consider to be the present disclosure, nor to indicate that the following experiments have been performed, or that they represent all experiments that may be performed. It should be understood that the exemplary descriptions described herein are not necessarily performed, but rather that by performing the descriptions, data on the properties described therein can be generated. Efforts have been made to ensure accuracy regarding the numbers used (e.g., quantity, temperature, etc.), but some experimental error and deviation should be taken into consideration.

[0259] Unless otherwise specified, parts are by weight, molecular weight is weight-average molecular weight, temperature is degrees Celsius (°C), and pressure is atmospheric pressure or near atmospheric pressure. Standard abbreviations are used, including: min = minute; h or hr = hour; equiv = equivalent; mg = milligram; g = gram; ml or mL = milliliter; l or L = liter; mM = millimoles; M = mole; HPLC = high-performance liquid chromatography; NMR = nuclear magnetic resonance; XRPD = X-ray powder diffraction; DSC = differential scanning calorimetry; DVS = dynamic vapor adsorption; RH = relative humidity; HPT = heptane; Depositphotos = ethyl acetate; EtOH = ethanol; DCM = dichloromethane; MTBE = methyl tert-butyl ether; MEK = methyl ethyl ketone.

[0260] LC: Agilent 1100 series; Mass spectrometer: Agilent G6120BA, single quad LC-MS method: Agilent Zorbax Eclipse Plus C18, 4.6 × 100 mm, 3.5 mM, 35°C, flow rate 1.5 mL / min, gradient from 0% to 100% B over 2.5 mins, washing at 100% B for 0.5 mins; A = 0.1% formic acid / 5% acetonitrile / 94.9% water; B = 0.1% formic acid / 5% water / 94.9% acetonitrile Flash column: ISCO Rf+ Reverse-phase HPLC: ISCO-EZ; Column: Kinetex 5mm EVO C18 100 A; 250×21.2mm (Phenomenex)

[0261] X-ray powder diffraction (XRPD) XRPD analysis was performed using PANalytical X'pert pro, scanning the sample at 3–35°²θ. The material was gently ground to release any weak aggregates, and the sample was supported by loading it into a multiwell plate with a Kapton or Mylar polymer film. The multiwell plate was then placed in a diffractometer and analyzed using Cu K irradiation (α1λ=1.54060Å, α2 / 1.54443Å, β=1.39225Å, α1:α2 ratio=0.5) in transmission mode (step size 0.0130°²θ) with a generator setting of 40kV / 40mA.

[0262] Differential Scanning Calorimetry (DSC) Approximately 5 mg of the material was weighed into an aluminum DSC pan and sealed non-airtightly with a perforated aluminum lid. The sample pan was then loaded into a Mettler Toledo DSC-3 and heated and held at 30°C until a stable heat flow response was obtained. Once a stable heat flow response was achieved, the sample and reference material were heated to 450°C at a scanning rate of 5°C / min, and the resulting heat flow response was monitored. Nitrogen was used as the purge gas, and a 50 cm³ filter was used. 3 It was used at a flow rate of [number] minutes.

[0263] Dynamic Vapor Adsorption (DVS) Approximately 10 mg of the sample was placed in a mesh vapor adsorption balance pan and loaded onto a Surface Measurement Systems DVS-1, DVS Intrinsic, or DVS Advantage dynamic vapor adsorption balance. The sample was subjected to a relative humidity (RH) gradient profile from 40 to 90% in 10% increments, with the sample maintained at 25°C at each step until a stable weight was achieved (dm / dt 0.004%, minimum step length 30 minutes, maximum step length 500 minutes). After the completion of the adsorption cycle, the sample was dried to 0% RH using the same procedure and then returned to 90% RH in a second adsorption cycle. Two cycles were performed. The weight change during the adsorption / desorption cycle was plotted to allow determination of the hygroscopicity of the sample. XRPD analysis was then performed on any held solid.

[0264] Example 1: Preparation of crystalline forms A and B of [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1S)-1-(2-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-b]pyridine-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid

[0265] [ka]

[0266] Step 1: Heterocyclic ammonium compound (25 g, 133 mmol) and ammonium sulfate (175 mg, 1 mol%) were packed into a 1 L round-bottom flask equipped with a magnetic stirring rod. HMDS (133 mL, 1 M) was added, and the mixture was refluxed under an air atmosphere for 4 hours (heating block temperature 155°C). Excess HMDS was evaporated under vacuum at 60°C, and the flask was then placed under high vacuum at 45°C for 30 minutes. This procedure was repeated to ensure that all excess HMDS was removed.

[0267] The pale orange oil residue was dissolved in anhydrous MeCN (266 mL), and sugar (46.55 g, 146.3 mmol) was added. The resulting mixture was stirred until all the sugar was dissolved (typically 5 minutes, resulting in a yellow solution). Then, TMSOTf (4.8 mL, 26.6 mmol) was added dropwise over 20 minutes (slight exothermic reaction). After the addition of TMSOTf was complete, LC-MS analysis showed that all the starting heterocycles had been consumed. The reaction mixture was then stirred for 17–20 hours (resulting in a darker mixture). LC-MS aliquots showed UV purity of over 90%, and the ratio between the desired product and its glycoside epimer was 97:3. Depositphotos (350 mL) and saturated NaHCO3 (300 mL) were added sequentially, at which point the mixture turned a deep blue color. The layers were separated, and the aqueous layer was extracted once with Depositphotos (150 mL). The combined organic layers were dried over Na2S2O3, filtered, and evaporated to dryness. The dark blue oil was dissolved in DCM (300 mL). Silica (50 g) and activated carbon (15 g) were added, and the resulting suspension was vigorously stirred for 1.5 hours. It was then filtered through Celite to obtain a clear pale yellow to colorless solution. The filtrate was evaporated to dryness to obtain the crude material. The clear oil was dissolved in siRNA (1.33 mL / g). The solution was vigorously stirred, and hexane (4.5 mL / g) was added, at which point a turbid mixture was obtained. The mixture was heated under reflux until completely dissolved, cooled to room temperature, and seed crystals were seeded. After 1 hour at room temperature, the mixture was placed in a refrigerator (0°C) for 20 hours. The crystals were then filtered and rinsed with cold MTBE (2 × 80 mL + 1 × 50 mL) to obtain the pure product (46.85 g, 79%). The mother liquor was evaporated to dryness, and the crystallization procedure was repeated. This yielded additional material (4.45g, 7%). The overall yield was 51.3g, 86%.

[0268] Step 2: A 5 L three-necked round-bottom flask was filled with a solution of the product from Step 1 (157 g, 353 mmol) in dimethyl sulfoxide (353 mL, 1 M). To this solution, (1S)-1-(2-fluorophenyl)ethylamine·HCl (93 g, 529 mmol, 1.5 equivalents), followed by triethylamine (170 mL, 1.2 mol, 3.5 equivalents). The reaction mixture was heated to 80 °C and stirred with an overhead mechanical stirrer for 48 hours. The mixture was cooled to room temperature and diluted with methanol (700 mL, 0.5 M). K2CO3 (233 g, 1.2 mol, 3.5 equivalents) was added, and the reaction mixture was stirred at room temperature. After 40 hours, the reaction mixture was filtered through Celite, and the filter cake was washed with methanol (2 × 200 mL). The solution was concentrated under vacuum to remove volatile substances. 3.5 L of water was added to the remaining solution with vigorous stirring. Next, the resulting precipitate was collected and washed with water (3 × 1 L) to obtain the desired product as a yellowish-brown solid (139 g, 92%).

[0269] Step 3: To a solution of the product obtained from Step 2 (45.74 g, 108 mmol) and 2,2-dimethoxypropane (66.3 ml, 541 mmol) in acetone (270 mL), p-TsOH (2.05 g, 10.8 mmol) was added at room temperature. The reaction mixture was stirred for 2 hours and then concentrated under reduced pressure. The amber-colored crude oil was reconstituted in siRNA (1.0 L) and washed with saturated NaHCO3 (500 mL). The organic layer was separated, stirred with activated carbon, and then filtered. The filtrate was filtered with Na2SO4 and concentrated under vacuum to obtain an off-white solid. The solid was suspended in a 1:1 siRNA:hexane (500 mL) and recovered by vacuum filtration. The filtration cake was washed with hexane (100 mL) and then dried under high vacuum to obtain the desired product as a white solid (39.2 g, 78%).

[0270] Step 4: To a suspension of methylenebis(phosphonic acid dichloride) (81.0 g, 324 mmol, 3.0 equivalents) in THF (162 mL, 2.0 M), N,N-diisopropylethylamine (20.7 mL, 119 mmol, 1.1 equivalents) was added at 0°C. To the resulting mixture, a solution of the product obtained from Step 3 (50.0 g, 108 mmol, 1.0 equivalent) in THF (347 mL, 0.31 M) was added dropwise over 1 hour. After the addition, the resulting mixture was stirred at 0°C for a further 15 minutes, and then the solution was transferred via cannula to a pre-cooled (0°C) flask containing 0.2 M HCl (1080 mL). The reaction mixture was heated to 30°C and stirred at 30°C for 16 hours [acetonide deprotection]. After completion, the reaction mixture was washed with 1.5:1 MTBE / THF (5 × 900 mL). The aqueous phase was diluted with brine (960 mL) and extracted with 2:1 2-MeTHF / THF (1 L). The organic phase was collected, and the aqueous phase was washed, then with brine (2 × 500 mL). DOWEX Marathon CH was added to the organic layer. + Form (20 g / L, 20 g) was added and stirred at room temperature for 2 hours. The DOWEX beads were removed by filtration, and the resulting solution was concentrated under reduced pressure to obtain a colorless / off-white foam (crude formula (I)).

[0271] To purify the compound by recrystallization, the solid was dissolved in EtOH (313 mL) with stirring, and then CH3CN (1,175 mL) was added over a period of 5 minutes. The resulting clear solution was stirred at 25°C for 1 hour, during which crystallization occurred. The mixture was allowed to stand at 25°C for 12 hours, and then the white solid was collected by vacuum filtration, rinsed with 6:1 CH3CN / EtOH (150 mL), and dried under reduced pressure at 55°C for 4 days to obtain the product as a white solid (32.6 g, yield 52%, UV purity 98.5%, containing 0.75 wt% CH3CN). The isolated solid was identified as crystalline form B by XRPD (see International Publication No. 2020 / 123772).

[0272] Next, when the sample of crystal morphology B was dried in a vacuum furnace at 60°C for 16 hours and determined by XRPD, it resulted in the isolation of crystal morphology A (see International Publication No. 2020 / 123772).

[0273] Example 2: Preparation of crystalline form I of formula (I) A flask was filled with Form A from Example 1 (1.00 g) and modified EtOH (5 mL, 5 parts). The resulting mixture was heated to approximately 55-60°C to form a first clear solution. Toluene (10.0 mL, 10 parts) was added at a temperature of 55-60°C to form a second clear solution. The second clear solution was cooled to room temperature over approximately 1.5 hours and stirred overnight (approximately 18 hours) at room temperature to form a white suspension. The white solid was recovered by vacuum filtration, rinsed with 1:1 EtOH / toluene (2 parts), and dried under vacuum at 55-60°C for 3 days to obtain Form I as a white solid (0.82 g, yield 82%, containing 0.04 wt% ethanol and approximately 0 wt% toluene).

[0274] Crystal morphology I was characterized by the XRPD pattern shown in Figure 1 and further characterized by the differential scanning calorimetry (DSC) thermogram shown in Figure 2.

[0275] Similar results are observed when using morphology B as the starting material.

[0276] Example 3: Alternative preparation of crystalline form I of formula (I) A flask was packed with 3.5 g of form A or form B and suspended in 17.5 mL of anhydrous ethanol. The reaction mixture was heated to 35–40°C to obtain a clear solution. Next, 35 mL of toluene was added to the solution in small amounts over 30 minutes while maintaining a temperature of 35–40°C. Subsequently, the mixture was cooled to 20–25°C, at which point the solution became a slurry. The slurry was stirred for 18 hours. The mixture was further cooled to 0–5°C and maintained at that temperature for 6 hours. The resulting solid was filtered through a Buchner funnel and dried by suction under N2 for 18 hours. The solid was identified as form I by XRPD.

[0277] Similar results are observed when using morphology B as the starting material.

[0278] Example 4: Preparation of crystalline form II of formula (I) A flask was filled with Form A from Example 1 (1.00 g) and modified EtOH (5 mL, 5 parts). The resulting mixture was heated to approximately 55-60°C for about 15 minutes to form a first clear solution. The first clear solution was cooled to room temperature over approximately 1.5-2 hours and stirred at room temperature for approximately 72 hours to form a white, highly viscous slurry suspension. Additional ethanol (2 parts) was added to the white, highly viscous slurry suspension, and the resulting suspension was reheated to approximately 55-60°C to form a second clear solution. The second clear solution was cooled to room temperature over approximately 1.5 hours and stirred at room temperature for approximately 2 hours to form a white, concentrated slurry suspension. The white solid was recovered by vacuum filtration, rinsed with EtOH (2 parts), and dried under vacuum at 55-60°C for 18 hours to obtain Form II as a white solid (0.37 g, yield 37%, containing 0.68 wt% ethanol).

[0279] Crystal morphology II was characterized by the XRPD pattern shown in Figure 3 and further characterized by the differential scanning calorimetry (DSC) thermogram shown in Figure 4.

[0280] Similar results are observed when using morphology B as the starting material.

[0281] Example 5: Preparation of crystalline form III of formula (I) A flask was filled with Form A from Example 1 (0.5 g) and modified EtOH (2.5 mL, 5 parts). The resulting mixture was heated to approximately 55-60°C to form a first clear solution. Methyl tert-butyl ether (5 mL, 10 parts) was added at a temperature of 55-60°C to form a second clear solution. The second clear solution was cooled to room temperature over approximately 1.5 hours and stirred overnight (approximately 18 hours) at room temperature to form a white suspension. The white solid was recovered by vacuum filtration, rinsed with 2:1 MTBE / EtOH (2 parts), and dried under vacuum at 55-60°C for 18 hours to obtain Form III as a white solid (0.34 g, yield 68%, containing approximately 0 wt% EtOH and 0.04 wt% methyl tert-butyl ether).

[0282] Crystal morphology III was characterized by the XRPD pattern shown in Figure 5 and further characterized by the differential scanning calorimetry (DSC) thermogram shown in Figure 6.

[0283] Similar results are observed when using morphology B as the starting material.

[0284] Example 6: Preparation of crystalline form IV of formula (I) A flask was filled with 3.0 g of Form A from Example 1 and 5 parts of anhydrous THF (15 mL). The resulting mixture was heated to about 55-60°C to form a first turbid solution. 4 parts of 12 mL of ethyl acetate were added at a temperature of 55-60°C to form a second turbid solution. The second turbid solution was cooled to 50-55°C (as disclosed in International Publication No. 2020 / 123772), and about 20 mg of crystals of Form A of compound (I) was sprinkled on it. The resulting mixture was cooled to room temperature for about 1 hour, and about 20 mg of crystals of Form A of compound (I) was sprinkled on it again. The resulting mixture was stirred at room temperature overnight (about 18 hours) to form an off-white suspension. The solid was recovered by vacuum filtration, rinsed with 2:1 THF / siRNA (6 mL, 2 parts), and dried under vacuum at 55-60°C for 18 hours to obtain Form IV as a white solid (2.8 g, yield 93%, containing 0.05 wt% THF and 0.38 wt% siRNA).

[0285] Crystal morphology IV was characterized by the XRPD pattern shown in Figure 7.

[0286] Similar results are observed when using morphology B as the starting material.

[0287] Example 7: Preparation of crystalline form V of formula (I) A flask was filled with Form B of Example 1 (1.00 g) and anhydrous EtOH (5 mL, 5 parts). The resulting slurry was heated to approximately 35°C and stirred for 7 hours to form a clear solution. The mixture was cooled to 20-25°C and stirred for a further 18 hours to obtain a white suspension. The white solid was recovered by vacuum filtration.

[0288] Crystal morphology V was characterized by XRPD as shown in Figure 8 and further characterized by differential scanning calorimetry (DSC) thermogram as shown in Figure 9. 1 1H NMR revealed the presence of 7.6 wt / wt% EtOH, indicating that form V is the EtOH solvate.

[0289] Next, it was found that morphology V slowly converted to morphology II when stored at ambient temperature under an N2 blanket (over 48 hours for complete conversion). When the temperature was raised to 40°C and morphology V was dried under vacuum, the conversion rate to morphology II increased (complete conversion was observed after 48 hours). When morphology II was resuspended overnight in a 1:1 EtOH:SiO mixture with stirring, morphology V could be recovered, demonstrating that the relationship between morphology II and morphology V is reversible.

[0290] Example 8: Preparation of Form VI A 20 mL vial was filled with 0.5 g of morphology B and 1.5–2.0 mL of EtOH. This suspension was vigorously stirred at 20–25°C for 3 days. The solid was recovered by filtration. Crystal morphology VI was characterized by XRPD (Figure 10) and DSC (Figure 11).

[0291] Example 9: Competitive Slurry Experiment Crystal morphology A and morphology I Crystal morphology A (1.5 g) and crystal morphology I (1.5 g) were packed into a flask with 30 mL of a 1:2 mixture of anhydrous ethanol and ethyl acetate. The resulting mixture was stirred at room temperature for 5 hours to obtain a white, high-viscosity slurry. 6 mL of a 1:2 mixture of anhydrous ethanol and ethyl acetate was then added. The suspension was stirred for 5 days. Small samples of the suspension were taken from the mixture at 22, 46, and 118 hours. The samples were filtered, dried in an oven at room temperature for 2-3 hours, and characterized by XRPD. The results indicated that morphology I was the final morphology after 46 hours of stirring, and that morphology I was more stable than morphology A under these conditions.

[0292] Crystal morphology I and morphology V at room temperature A 20 mL vial was filled with 0.2 g of form V and 0.2 g of form I, along with 4 mL of a 1:2 EtOH (anhydrous): Depositphotos mixture. The resulting suspension was vigorously stirred for 3 days. The suspension was filtered, and the solid was dried at room temperature under reduced pressure for 3 hours. The isolated solid was identified as form I by XRPD, and form I was shown to be more stable than form V under these conditions.

[0293] Crystal morphology I and morphology V at 35°C A 15 mL round-bottom flask was fitted with a stirring rod, thermometer, and N2 inlet, and filled with 0.25 g of Form I and 0.25 g of Form V. 6 mL of a 1:2 EtOH (anhydrous):HCl mixture was added to the flask and mixed to obtain a white suspension. The suspension was heated to 35°C and vigorously stirred for 2 days. The suspension was filtered, and the resulting solid was dried at room temperature under reduced pressure for 3 hours. The isolated solid was identified as Form I by XRPD, and Form I was shown to be more stable than Form V under these conditions.

[0294] Crystal morphologies I and II at room temperature A 20 mL vial was filled with 0.2 g of Form I and 0.2 g of Form II. 4 mL of a 1:2 EtOH (anhydrous):alkyl mixture was added to the vial to obtain a homogeneous suspension. The suspension was vigorously stirred at room temperature for 5 days. The suspension was filtered, and the resulting solid was dried at room temperature under reduced pressure for 3 hours. The isolated solid was identified as Form I by XRPD, demonstrating that Form I is more stable than Form II under these conditions.

[0295] Crystal morphology I and II at 35°C A 15 mL round-bottom flask was fitted with a stirring rod, thermometer, and N2 inlet, and packed with 0.2 g of Form I and 0.2 g of Form II. 5 mL of a 1:2 EtOH (anhydrous):HCl mixture was added to the flask and mixed to obtain a white suspension. The suspension was heated to 35 °C and vigorously stirred for 2 days. The suspension was filtered, and the resulting solid was dried at room temperature under reduced pressure for 3 hours. The isolated solid was identified as Form I by XRPD, and Form I was shown to be more stable than Form II under these conditions.

[0296] The results of the competitive slurry experiments are summarized in Table 2 below.

[0297] [Table 2]

[0298] Example 10. Competitive slurries of crystalline forms I, II, and V at room temperature The relative morphological stability between morphologies I, II, and V was investigated by subjecting mixtures of the mixed morphologies to changes over time in EtOH, SiO2, and mixtures thereof. The morphology V starting material contained some morphology II, as determined by its XRPD pattern.

[0299] In two 4 mL vials, sufficient amounts of morphology I and morphology V (containing some morphology II) were mixed with 1 mL of a suitable solvent to produce slurries in each vial. The two slurries were combined and shaken at 600 rpm at 20°C. The solid obtained from the slurry was isolated for XRPD characterization at 0 hours, 1 day, and 2 days (Figure 12a-d). The results are summarized in Table 3 below.

[0300] [Table 3] * Two days later, slow response.

[0301] The XRPD patterns of the final solid recovered after overnight aging are shown in Figures 12a-d. It can be confirmed that both forms I and II were converted to form II in solvents containing 80% or more EtOH, while the mixture of forms I and II was retained in a 67% EtOH system. In 50-33% EtOH solvents, form V was converted to form I, but in EtOH systems of 20% or less, forms I and V were maintained after two 2-day aging cycles. The lack of conversion in EtOH systems of 20% or less was attributed to the low solubility of the crystalline forms.

[0302] Example 11. Solubility profiles of forms I, II, and V in EtOH / SiOT The equilibrium solubility of forms I and V was measured at 20°C in different EtOH / SiO2 ratios. Form V starting material contains some form II, as determined by XRPD.

[0303] In 4 mL vials, 50 mg of a suitable solid was mixed with 1 mL of each solvent. The resulting slurry was shaken overnight at 600 rpm at 20°C. The supernatant obtained from each vial was sampled for HPLC to determine equilibrium solubility, the solid was isolated, and characterized by XRPD. The results are plotted in Figure 13 and summarized in Table 4 below.

[0304] [Table 4]

[0305] The solubility profile indicates that ethanol increases the solubility of forms I and V, while siRNA is a poor solvent. A maximum solubility of approximately 46 mg / mL was observed for form I in pure EtOH. No change in crystal morphology was observed in the vial of form I. In the vial containing a mixture of forms II and V, form II was the final form in the high ethanol (50% by volume or more) system. At low ethanol content (less than 50% by volume), no change in morphology was observed in the vial containing a mixture of forms II and V.

[0306] The data supports the following: a) Form I is more stable than Forms II and V in dimethyl-rich solvent systems, except for excessively high levels of dimethyl (excessively low solubility); and b) Form II is more stable than Forms I and V in solvent systems that are rich in EtOH or dilute in dimethyl.

[0307] Example 12: Polymorphism relationship map based on the EtOH:SiO solvent system A map of the polymorphism relationships between crystal forms I, II, and V was inferred from observations using an EtOH:HCl solvent system (Figure 14). From the EtOH:HCl solvent system, it was found that a) a solvent system rich in HCl is preferable for isolating form I, and b) a solvent system dilute or lacking HCl is preferable for isolating form II.

[0308] Example 13: Solubility profiles of forms I, II, and V in EtOH:heptane The equilibrium solubility of forms I, II, and V was measured in EtOH / heptane (HPT) solutions of different proportions at 20 and 35°C.

[0309] In 4 mL test vials, a suitable solid (Form II containing Form I or Form V) was mixed with approximately 1 mL of the respective solvent to produce a slurry. The resulting slurry was shaken overnight at 600 rpm at 20 and 35 °C. After overnight aging, the supernatant obtained from each vial was sampled for HPLC to determine equilibrium solubility, and the solid was isolated for identification by XRPD.

[0310] Figure 15 plots the solubility profiles of form I, as well as mixtures of forms II and V, in EtOH / HPT at 20 and 35°C. HPT is shown as an effective poor solvent. The solubility of form I is higher than that of the form II / V mixture at all solvent ratios, indicating that form II / V is the most stable form under these conditions. The large solubility difference between form I and the mixtures of forms II and V under EtOH-rich conditions means that isolating form II / V from EtOH / HPT is easier than isolating form I.

[0311] Example 14. DVS Characterization of Morphologies I and II Morphologies I and II were analyzed by dynamic vapor adsorption (DVS) at 25°C (Figures 16 and 17). Morphology II showed water uptake at a much lower relative humidity (RH) of 40%–70% than morphology I. XRPD characterization of the samples after DVS indicated that both morphologies underwent conversion to morphology VI during the experiment.

[0312] While the foregoing disclosures are described in some detail as explanations and examples for clarity of understanding, it will be understood by those skilled in the art that certain changes and modifications may be made within the scope of the attached claims. In addition, each of the references provided herein is incorporated by reference to the same extent as each of the references is incorporated by reference individually. In the event of any conflict between this application and the references provided herein, this application shall prevail. The present invention provides, for example, the following items: (Item 1) Equation (I): [ka] Crystal morphology I of the compound, characterized by an X-ray powder diffraction (XRPD) pattern containing three or more peaks at 11.1, 11.6, 13.8, 14.7, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). (Item 2) Crystal morphology I as described in item 1, characterized by an XRPD pattern containing five or more peaks at 11.1, 11.6, 13.8, 14.7, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). (Item 3) Crystal morphology I as described in item 1, characterized by an XRPD pattern containing seven or more peaks at 11.1, 11.6, 13.8, 14.78, 15.4, 16.6, 17.0, 18.6, 19.3, 20.1, 21.3, 22.1, 23.0, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). (Item 4) Crystal morphology I as described in item 1, characterized by an XRPD pattern containing peaks at 11.1, 13.8, 18.6, 20.1, 23.0, and 24.8 degrees 2θ (±0.2 degrees 2θ). (Item 5) Crystal morphology I as described in item 4, wherein the XRPD pattern further includes one or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). (Item 6) Crystal morphology I as described in item 4, wherein the XRPD pattern further includes three or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). (Item 7) Crystal morphology I as described in item 4, wherein the XRPD pattern further includes five or more peaks at 11.6, 14.7, 15.4, 16.6, 17.0, 19.3, 21.3, 22.1, 24.8, 26.6, 27.3, and 29.1 degrees 2θ (±0.2 degrees 2θ). (Item 8) The aforementioned X-ray powder diffraction pattern is substantially as shown in Figure 1, wherein the crystal morphology I described in item 1. (Item 9) Crystalline form I as described in item 1, substantially free from other crystalline or amorphous forms of the compound of formula (I). (Item 10) Crystal morphology I as described in any one of items 1 to 9, further characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at approximately 163.9°C. (Item 11) Crystal morphology I as described in any one of items 1 to 9, further characterized by a melting point onset of approximately 155.1°C, as determined by differential scanning calorimetry thermogram (DSC). (Item 12) Crystal morphology I as described in item 10 or 11, wherein the DSC thermogram is substantially as shown in Figure 2. (Item 13) Equation (I): [ka] Crystallographic form II of the compound, characterized by an X-ray powder diffraction (XRPD) pattern containing three or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). (Item 14) Crystal morphology II as described in item 13, characterized by an XRPD pattern containing five or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). (Item 15) Crystal morphology II as described in item 13, characterized by an XRPD pattern containing seven or more peaks at 10.1, 10.8, 12.8, 13.7, 16.5, 17.7, 19.0, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). (Item 16) Crystal morphology II as described in item 13, characterized by an XRPD pattern containing peaks at 16.5, 22.8, and 24.6 degrees 2θ (±0.2 degrees 2θ). (Item 17) Crystal morphology II as described in item 16, wherein the XRPD pattern further includes one or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ). (Item 18) Crystal morphology II as described in item 16, wherein the XRPD pattern further includes three or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ). (Item 19) Crystal morphology II as described in item 16, wherein the XRPD pattern further includes five or more peaks at 10.1, 10.8, 12.8, 13.7, 17.7, and 19.0 degrees 2θ (±0.2 degrees 2θ). (Item 20) Crystal morphology II as described in item 13, wherein the X-ray powder diffraction pattern is substantially as shown in Figure 3. (Item 21) Crystalline form II as described in item 13, substantially free from other crystalline or amorphous forms of the compound of formula (I). (Item 22) Crystal morphology II as described in any one of items 13-21, further characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at approximately 166.5°C. (Item 23) Crystal morphology II as described in any one of items 13-22, further characterized by a melting point onset of approximately 157.4°C, as determined by differential scanning calorimetry thermogram (DSC). (Item 24) Crystal morphology II as described in item 22 or 23, wherein the DSC thermogram is substantially as shown in Figure 4. (Item 25) Equation (I): [ka] Crystallographic morphology III of the compound, characterized by an X-ray powder diffraction (XRPD) pattern containing three or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). (Item 26) Crystal morphology III as described in item 25, characterized by an XRPD pattern containing five or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). (Item 27) Crystal morphology III as described in item 25, characterized by an XRPD pattern containing seven or more peaks at 6.6, 10.9, 14.2, 16.1, 18.4, 19.3, 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). (Item 28) Crystal morphology III as described in item 25, characterized by an XRPD pattern containing peaks at 6.6, 10.9, 14.2, 16.1, 18.4, and 19.3 degrees 2θ (±0.2 degrees 2θ). (Item 29) Crystal morphology III as described in item 28, wherein the XRPD pattern further includes one or more peaks at 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). (Item 30) Crystal morphology III as described in item 28, wherein the XRPD pattern further includes three or more peaks at 20.2, 22.0, 24.7, and 28.1 degrees 2θ (±0.2 degrees 2θ). (Item 31) Crystal morphology III as described in item 25, wherein the X-ray powder diffraction pattern is substantially as shown in Figure 5. (Item 32) Crystalline form III as described in item 25, substantially free from other crystalline or amorphous forms of the compound of formula (I). (Item 33) Crystal morphology III as described in any one of items 25-32, further characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at approximately 161.8°C. (Item 34) Crystal morphology III as described in any one of items 25-32, further characterized by a melting point onset of approximately 149.6°C, as determined by differential scanning calorimetry thermogram (DSC). (Item 35) Crystal morphology III as described in item 33 or 34, wherein the DSC thermogram is substantially as shown in Figure 6. (Item 36) Equation (I): [ka] Crystallographic morphology IV of the compound, characterized by an X-ray powder diffraction (XRPD) pattern containing three or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). (Item 37) Crystal morphology IV as described in item 36, characterized by an XRPD pattern containing five or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). (Item 38) Crystal morphology IV as described in item 36, characterized by an XRPD pattern containing seven or more peaks at 6.0, 11.2, 14.1, 17.0, 19.5, 23.2, 25.1, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). (Item 39) Crystal morphology IV as described in item 36, characterized by an XRPD pattern containing peaks at 14.1, 17.0, 19.5, 23.2, and 25.1 degrees 2θ (±0.2 degrees 2θ). (Item 40) Crystal morphology IV as described in item 39, wherein the XRPD pattern further includes one or more peaks at 6.0, 11.2, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). (Item 41) Crystal morphology IV as described in item 39, wherein the XRPD pattern further includes three or more peaks at 6.0, 11.2, 27.1, and 28.9 degrees 2θ (±0.2 degrees 2θ). (Item 42) Crystal morphology IV described in item 36, wherein the X-ray powder diffraction pattern is substantially as shown in Figure 7. (Item 43) Crystalline form IV as described in item 36, substantially free from other crystalline or amorphous forms of the compound of formula (I). (Item 44) Equation (I): [ka] Crystal morphology V of the compound, characterized by an X-ray powder diffraction (XRPD) pattern containing three or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.5, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ). (Item 45) Crystal morphology V as described in item 44, characterized by an XRPD pattern containing five or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.5, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ). (Item 46) Crystal morphology V as described in item 44, characterized by an XRPD pattern containing seven or more peaks at 10.4, 15.1, 15.8, 16.3, 16.8, 18.5, 19.1, 19.7, 21.7, 22.1, 23.0, 23.5, 26.0, 26.5, 28.4, 28.9, and 31.4 degrees 2θ (±0.2 degrees 2θ). (Item 47) Crystal morphology V, as described in item 44, characterized by an XRPD pattern containing peaks at 15.8, 16.3, 16.8, 18.5, 19.1, 21.7, 22.1, and 23.0 degrees 2θ (±0.2 degrees 2θ). (Item 48) Crystal morphology V as described in item 47, wherein the XRPD pattern further includes one or more peaks at 10.4, 15.1, 19.7, and 23.6 degrees 2θ (±0.2 degrees 2θ). (Item 49) Crystal morphology V as described in item 47, wherein the XRPD pattern further includes three or more peaks at 10.4, 15.1, 19.7, and 23.6 degrees 2θ (±0.2 degrees 2θ). (Item 50) The aforementioned X-ray powder diffraction pattern is substantially as shown in Figure 8, for the crystal morphology V described in item 44. (Item 51) Crystalline form V as described in any one of items 44 to 50, substantially free from other crystalline or amorphous forms of the compound of formula (I). (Item 52) Crystal morphology V as described in any one of items 44-51, further characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at approximately 150.4°C. (Item 53) Crystal morphology V as described in any one of items 44-52, further characterized by a melting point onset of approximately 135.8°C, as determined by differential scanning calorimetry thermogram (DSC). (Item 54) Crystal morphology V as described in item 52 or 53, wherein the DSC thermogram is substantially as shown in Figure 9. (Item 55) Equation (I): [ka] Crystallographic form VI of the compound, characterized by an X-ray powder diffraction (XRPD) pattern containing three or more peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, 24.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). (Item 56) Crystal morphology VI as described in item 55, characterized by an XRPD pattern containing five or more peaks at 5.8, 10.4, 16.2, 19.4, 21.3, 22.4, 24.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). (Item 57) Crystal morphology VI as described in item 55, characterized by an XRPD pattern containing peaks at 19.4, 21.3, 22.4, and 24.4 degrees 2θ (±0.2 degrees 2θ). (Item 58) Crystal morphology VI as described in item 57, wherein the XRPD pattern further includes one or more peaks at 5.8, 10.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). (Item 59) Crystal morphology VI as described in item 57, wherein the XRPD pattern further includes three or more peaks at 5.8, 10.4, 27.5, and 31.1 degrees 2θ (±0.2 degrees 2θ). (Item 60) Crystal morphology VI as described in any one of items 55 to 59, wherein the X-ray powder diffraction pattern is substantially as shown in Figure 10. (Item 61) Crystalline form VI as described in any one of items 55 to 60, substantially free from other crystalline or amorphous forms of the compound of formula (I). (Item 62) Crystal morphology VI as described in any one of items 55-61, further characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at approximately 142.9°C. (Item 63) Crystal morphology VI as described in any one of items 55-62, further characterized by a melting point onset of approximately 116.6°C, as determined by differential scanning calorimetry thermogram (DSC). (Item 64) Crystal morphology VI as described in item 62 or 63, wherein the DSC thermogram is substantially as shown in Figure 11. (Item 65) A pharmaceutical composition comprising a crystalline form described in any one of items 1 to 64 and a pharmaceutically acceptable carrier. (Item 66) A method for treating a disease, disorder, or condition at least partially mediated by CD73, wherein the method comprises administering an effective amount of a crystalline form of a compound described in any one of items 1 to 64 to a subject in need. (Item 67) The method according to item 66, wherein the compound is administered in an amount effective to reverse, halt, or delay the progression of CD73-mediated immunosuppression. (Item 68) The method according to item 66, wherein the disease, disorder, or condition is cancer. (Item 69) The method according to item 68, wherein the cancer is cancer of the prostate, colon, rectum, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovaries, testes, head, neck, skin (including melanoma and basal cell carcinoma), mesothelial layer, white blood cells (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell lung cancer and non-small cell lung cancer), adrenal gland, thyroid, kidney, or bone, or is glioblastoma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma (including Kaposi's sarcoma), choriocarcinoma, cutaneous basal cell carcinoma, or testicular seminoma. (Item 70) The method according to item 68, wherein the cancer is selected from the group consisting of melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia, brain tumor, lymphoma, ovarian cancer, and Kaposi's sarcoma. (Item 71) The method according to item 66, wherein the disease, disorder, or condition is an immune-related disease, disorder, or condition selected from the group consisting of rheumatoid arthritis, renal failure, lupus, asthma, psoriasis, colitis, pancreatitis, allergy, fibrosis, anemic fibromyalgia, Alzheimer's disease, congestive heart failure, stroke, aortic stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infection, Crohn's disease, ulcerative colitis, allergic contact dermatitis and other eczema, systemic sclerosis, and multiple sclerosis. (Item 72) A combination comprising a crystalline form of a compound described in any one of items 1 to 64 and at least one additional therapeutic agent. (Item 73) The combination described in item 72, wherein the at least one additional therapeutic agent is a chemotherapeutic agent, an immunomodulatory agent and / or an anti-inflammatory agent, an anti-hypercholesterolemia agent, an anti-infective agent, or radiation therapy. (Item 74) The combination described in item 72, wherein at least one of the additional therapeutic agents is an immune checkpoint inhibitor. (Item 75) The combination described in item 74, wherein the immune checkpoint inhibitor is selected from the group consisting of PD-1, PD-L1, BTLA, LAG-3, B7 family members, TIM-3, TIGIT, and CTLA-4. (Item 76) The combination described in item 73, wherein the at least one additional therapeutic agent is a chemotherapeutic agent selected from the group consisting of gemcitabine, nab-paclitaxel, enzalutamide, doxorubicin, or any combination thereof. (Item 77) The combination described in item 73, wherein at least one additional therapeutic agent is an immunomodulator that modulates the levels of adenosine or arginase. (Item 78) A method for treating cancer in a subject, wherein the method comprises administering to the subject an effective amount of a crystalline form of a compound described in any one of items 1 to 64 and at least one additional therapeutic agent. (Item 79) The method according to item 78, wherein the at least one additional therapeutic agent is a chemotherapeutic agent, an immunomodulatory agent and / or an anti-inflammatory agent, an anti-hypercholesterolemia agent, an anti-infective agent, or radiation therapy. (Item 80) The method according to item 78, wherein the at least one additional therapeutic agent is an immune checkpoint inhibitor. (Item 81) The method according to item 80, wherein the at least one additional therapeutic agent is an immune checkpoint inhibitor selected from the group consisting of PD-1, PD-L1, BTLA, LAG-3, B7 family members, TIM-3, TIGIT, and CTLA-4. (Item 82) The method according to item 79, wherein the at least one additional therapeutic agent is a chemotherapeutic agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, enzalutamide, docetaxel, nab-paclitaxel, gemcitabine, and doxorubicin. (Item 83) The method according to item 79, wherein the at least one additional therapeutic agent is an immunomodulator that modulates the levels of adenosine or arginase. (Item 84) The method according to item 78, wherein the compound and the at least one additional therapeutic agent are administered in combination. (Item 85) The method according to item 78, wherein the compound and the at least one additional therapeutic agent are administered sequentially. (Item 86) The method according to item 85, wherein the compound is administered after the at least one additional therapeutic agent. (Item 87) The method according to item 85, wherein the compound is administered before the at least one additional therapeutic agent. (Item 88) The method according to item 78, wherein the treatment periods of the compound and the at least one additional therapeutic agent overlap. (Item 89) The method according to item 80, wherein the immune checkpoint inhibitor is zimbererimab or dombanarimab.

Claims

[Claim 1] The invention described herein.

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