CD73 inhibitor alkaline salt, preparation method therefor, and use thereof

Abstract

Disclosed are a CD73 inhibitor alkaline salt, a preparation method therefor, and a use thereof. The CD73 inhibitor is a compound having the structure shown in formula (I). An alkaline salt of the compound shown in the formula (I) can greatly improve physical and chemical properties such as the chemical stability, solubility, and hygroscopicity of an amorphous free acid of the compound represented by formula (I), meet industrial production requirements, and satisfy development requirements for clinical medicine preparations. The alkaline salt of the compound represented by formula (I) can be widely applied in the preparation of drugs for treating tumors, immune-related diseases, and metabolic diseases mediated at least in part by CD73.

Description

A CD73 inhibitor basic salt and its preparation method and application Technical Field

[0001] The present invention belongs to the field of drug development, and specifically relates to a basic salt of a CD73 inhibitor, a preparation method, and an application thereof. Background Art

[0002] CD73, also known as extracellular 5'-nucleotidase (Ecto-5'-nucleotidase, eNT), is a 70kDa protein molecule. Under normal circumstances, it is expressed on vascular endothelial cells and some blood cells. It is anchored to the cell membrane surface through glycosylphosphatidylinositol (GPI) and regulates the metabolism of adenosine triphosphate (ATP) together with CD39. Among them, CD39 (also known as extracellular nucleoside diphosphate hydrolase-NTPDase1) can catalyze ATP to generate adenosine monophosphate (AMP), producing only a small amount of adenosine diphosphate (ADP). The main function of CD73 is to catalyze the conversion of extracellular nucleotides (such as 5'AMP) into their corresponding nucleosides (such as adenosine).

[0003] Nucleosides, particularly adenosine, produced by CD73 are considered endogenous regulators of a wide range of physiological functions. Adenosine regulates the cardiovascular system, central nervous system, respiratory system, kidneys, adipocytes, platelets, and the immune system. Within the immune system, extracellular adenosine can act on a wide variety of immune cells and mediate anti-inflammatory responses. In many tissues, adenosine also promotes fibrosis.

[0004] CD73 expression has been found in many tumor cell types, including leukemia, bladder cancer, glioma, glioblastoma, ovarian cancer, melanoma, prostate cancer, thyroid cancer, esophageal cancer, and breast cancer. Furthermore, CD73 expression has been found on the surface of immunosuppressive cells, including regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs). High CD73 expression has also been associated with angiogenesis, invasion, chemotherapy resistance, metastasis, and shorter survival in various tumor types, including breast cancer and melanoma.

[0005] Mechanistic studies have shown that malignant tumor cells release large amounts of ATP under chemotherapy and other stressors, which is rapidly converted into adenosine and accumulates in the tumor microenvironment. The release of extracellular ATP due to cell death or intracellular stress activates the immune response, but the ATP metabolite adenosine has immunosuppressive activity. A key point is that adenosine in the tumor inhibits infiltrating effector T lymphocytes by activating adenosine receptors (such as A2A), thereby promoting tumor development. Therefore, the accumulation of extracellular adenosine in tumor tissue is an important mechanism for tumor immune escape.

[0006] Reducing CD73 expression with interfering RNA or overexpressing CD73 in tumor cells can modulate tumor growth and migration. CD73 knockout mice are less susceptible to organ transplant rejection and spontaneous tumor development. Genetic deletion of the A2A receptor gene can induce T cell-dependent tumor rejection. In mouse models, treatment with an antibody that binds to mouse CD73 inhibits breast tumor growth and migration.

[0007] Therefore, targeting CD73 represents a potential therapeutic strategy that can enhance the efficacy of anti-tumor therapy and provide a new therapeutic strategy for limiting further tumor progression. At the same time, targeting CD73 can also be used to treat other diseases mediated by adenosine, such as enhancing immune responses, enhancing immune effects, enhancing inflammatory responses, and treating neurological disorders, neurodegenerative diseases, and central nervous system diseases, such as depression, Parkinson's disease, sleep disorders, fibrosis, and other immune-inflammatory diseases.

[0008] Shanghai Abbisko Therapeutics Co., Ltd. has discovered a small molecule compound (WO2018113584A1) with CD73 inhibitory effects during its long-term research. Representative compounds are as follows:

[0009] The Chinese name is: (((((2R,3S,4R,5R)-5-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphino)methyl)phosphonic acid (compound of formula (I)). The compound has a strong inhibitory effect on CD73 enzymatic and cytological activity and can be used to treat diseases that are at least partially mediated by CD73. Tumors, immune-related diseases, metabolic diseases, such as prostate cancer, colon cancer, rectal cancer, pancreatic cancer, gastric cancer, endometrial cancer, cervical cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer, neck cancer, skin cancer, mesothelial lining cancer, white blood cell cancer, esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, lung cancer, adrenal cancer, thyroid cancer, kidney cancer, bone cancer, brain tumor, glioblastoma, mesothelioma, renal cell carcinoma, sarcoma, choriocarcinoma, epidermal basal cell carcinoma, testicular seminoma, etc.

[0010] The applicant obtained the free acid lyophilized powder of the compound of formula (I) according to the preparation method disclosed in the specification, which was identified as an amorphous form. During the subsequent pharmaceutical research process, it was found that its physicochemical properties were poor and unstable, and it could not meet the needs of subsequent clinical formulation development and industrial production.

[0011] Therefore, in order to meet the needs of clinical research and marketed pharmaceutical preparations, it is urgent to develop an aggregated form of the compound of formula (I) that is suitable for drug development to overcome the defects of the existing technology. Summary of the Invention

[0012] To address the problems of the prior art, the inventors conducted in-depth research on the different aggregation forms of the compound of formula (I) and developed several basic salts of the compound of formula (I). These salts significantly improved the physicochemical properties of the free form of the compound of formula (I), including chemical stability, solubility, and hygroscopicity. The basic salts of the compound of formula (I) meet the requirements of industrial production and can meet the needs of clinical drug formulation development. Furthermore, during the salt screening process, the inventors unexpectedly discovered several crystalline free acids of the compound of formula (I), which exhibited excellent stability and hygroscopicity and could be used for medium- and long-term storage in industrial production processes or as raw materials for basic salt production. These basic salts and crystalline free acids of the compound of formula (I) possess significant clinical application value and are expected to accelerate their development into a new generation of small molecule CD73 inhibitors.

[0013] The first aspect of the present invention provides a basic salt of a compound of formula (I),

[0014] Wherein, the basic salt is sodium salt, potassium salt, calcium salt, arginine salt, meglumine salt or ammonium salt.

[0015] As a preferred solution, the basic salt is a sodium salt or a calcium salt.

[0016] As a further preferred embodiment, the molar ratio of the compound of formula (I) to the base in each molecule of the basic salt of the compound of formula (I) is 1:1 to 1:5.

[0017] As a further preferred embodiment, the molar ratio of the compound of formula (I) to the base in each molecule of the basic salt of the compound of formula (I) is 1:1, 1:2, 1:3, 1:4 or 1:5.

[0018] As a further preferred embodiment, the basic salt of the compound of formula (I) is the sodium salt of the compound of formula (I), and the molar ratio of the compound of formula (I) to sodium atoms in each molecule of the sodium salt of the compound of formula (I) is 1:1, 1:3 or 1:5.

[0019] As a further preferred embodiment, the molar ratio of the compound of formula (I) to sodium atoms in each molecule of the sodium salt of the compound of formula (I) is 1:3.

[0020] As a preferred embodiment, the sodium salt of the compound of formula (I) is an amorphous or crystalline compound.

[0021] As a preferred embodiment, the basic salt of the compound of formula (I) is the trisodium salt of the amorphous form of the compound (I).

[0022] As a preferred embodiment, the basic salt of the compound of formula (I) is a crystalline trisodium salt of the compound of formula (I).

[0023] As a further preferred embodiment, the crystalline form of the trisodium salt of the compound of formula (I) is Form A', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.22±0.2°, 6.38±0.2°, 9.55±0.2°, 12.72±0.2°, 19.10±0.2°, 20.01±0.2°, 20.53±0.2°, 23.13±0.2°, 25.50±0.2°, 26.27±0.2°, 26.66±0.2°, 28.80±0.2°, 29.43±0.2°, 32.06±0.2° and 35.37±0.2°.

[0024] As the most preferred embodiment, the X-ray powder diffraction pattern of the crystalline form A' includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG27 , and its X-ray powder diffraction data are shown in the following table:

[0025] This crystalline form of the trisodium salt of the compound of formula (I) is designated Form A'.

[0026] As a further preferred embodiment, the crystalline form of the trisodium salt of the compound of formula (I) is form B', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.15±0.2°, 6.27±0.2°, 9.34±0.2°, 12.34±0.2°, 12.59±0.2°, 15.64±0.2°, 17.06±0.2°, 18.90±0.2°, 23.60±0.2° and 25.93±0.2°.

[0027] As the most preferred embodiment, the X-ray powder diffraction pattern of the crystalline form B' includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG40 , and its X-ray powder diffraction data are shown in the following table:

[0028] This crystalline form of the trisodium salt of the compound of formula (I) is designated Form B'.

[0029] As a further preferred embodiment, the crystalline form of the trisodium salt of the compound of formula (I) is Form C', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 11.4±0.2°, 17.09±0.2°, 17.68±0.2°, 19.01±0.2°, 19.59±0.2°, 19.97±0.2°, 20.81±0.2°, 21.65±0.2°, 22.39±0.2°, 22.81±0.2°, 24.80±0.2°, 27.06±0.2°, 27.73±0.2°, 30.53±0.2° and 32.62±0.2°.

[0030] As the most preferred embodiment, the X-ray powder diffraction pattern of the crystalline form C' includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG44 , and its X-ray powder diffraction data are shown in the following table:

[0031] This crystalline form of the trisodium salt of the compound of formula (I) is designated Form C'.

[0032] As a further preferred embodiment, the crystalline form of the trisodium salt of the compound of formula (I) is Form D', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 11.97±0.2°, 17.97±0.2°, 18.92±0.2°, 19.41±0.2°, 20.36±0.2°, 20.97±0.2°, 21.66±0.2°, 22.88±0.2°, 23.24±0.2°, 24.15±0.2°, 24.77±0.2°, 25.11±0.2°, 25.51±0.2°, 26.21±0.2° and 30.10±0.2°.

[0033] As the most preferred embodiment, the X-ray powder diffraction pattern of the crystalline form D' includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG31 , and its X-ray powder diffraction data are shown in the following table:

[0034] This crystalline form of the trisodium salt of the compound of formula (I) is designated Form D'.

[0035] As a further preferred embodiment, the crystalline form of the trisodium salt of the compound of formula (I) is form E', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.15±0.2°, 3.24±0.2°, 9.79±0.2°, 12.35±0.2°, 12.41±0.2°, 14.30±0.2°, 18.61±0.2°, 19.91±0.2°, 20.80±0.2°, 23.94±0.2°, 24.89±0.2°, 31.39±0.2° and 34.67±0.2°.

[0036] As the most preferred embodiment, the X-ray powder diffraction pattern of the crystalline form E' includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG32 , and its X-ray powder diffraction data are shown in the following table:

[0037] This crystalline form of the trisodium salt of the compound of formula (I) is designated Form E'.

[0038] As a further preferred embodiment, the crystalline form of the trisodium salt of the compound of formula (I) is form F', and its X-ray powder diffraction pattern (XRPD) includes peaks at diffraction angles (2θ) of 3.12±0.2°, 5.81±0.2°, 6.27±0.2°, 9.43±0.2°, 18.94±0.2° and 25.58±0.2°.

[0039] As the most preferred embodiment, the X-ray powder diffraction pattern of the crystalline form F' includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG45 , and its X-ray powder diffraction data are shown in the following table:

[0040] This crystalline form of the trisodium salt of the compound of formula (I) is designated Form F'.

[0041] As a further preferred embodiment, the crystalline form of the trisodium salt of the compound of formula (I) is form A1', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.19±0.2°, 9.55±0.2°, 12.71±0.2°, 13.52±0.2°, 14.43±0.2°, 16.37±0.2°, 19.07±0.2°, 19.99±0.2°, 20.48±0.2°, 24.02±0.2°, 25.06±0.2°, 25.43±0.2°, 26.21±0.2°, 26.60±0.2° and 29.40±0.2°.

[0042] As the most preferred embodiment, the X-ray powder diffraction pattern of the crystalline form A1' includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG36 , and its X-ray powder diffraction data are shown in the following table:

[0043] The crystalline form of the trisodium salt of the compound of formula (I) is designated Form A1'.

[0044] As a further preferred embodiment, the basic salt of the compound of formula (I) is a calcium salt of the compound of formula (I), and the molar ratio of the compound of formula (I) to calcium atoms per molecule in the calcium salt of the compound of formula (I) is 1:1.

[0045] The second aspect of the present invention provides a method for preparing a basic salt of a compound of formula (I), comprising the following steps:

[0046] 1) dissolving or dispersing the free acid of the compound of formula (I) in water or an organic solvent, and adding an alkaline solution to the above system to carry out a salt-forming reaction; alternatively, adding the free acid of the compound of formula (I) to an alkaline solution to carry out a salt-forming reaction;

[0047] 2) collecting the solid product basic salt of the compound of formula (I) precipitated during the above-mentioned salt-forming reaction, or obtaining the solid product basic salt of the compound of formula (I) by creating supersaturation in the salt-forming system;

[0048] The free acid of the compound of formula (I) is an anhydrate, a hydrate or a solvate;

[0049] The basic salt of the compound of formula (I) is a sodium salt, potassium salt, calcium salt, arginine salt, meglumine salt or ammonium salt of the compound of formula (I);

[0050] The alkaline solution is selected from an aqueous solution or an organic solvent solution of sodium hydroxide, potassium hydroxide, calcium hydroxide, arginine, meglumine or ammonia.

[0051] The third aspect of the present invention provides a method for preparing a basic salt of a compound of formula (I), comprising the following steps:

[0052] 1) dissolving or dispersing the pentasodium salt of the compound of formula (I) in water or an organic solvent;

[0053] 2) stirring or suspending, and collecting the trisodium salt solid of the compound of formula (I) precipitated in the above process.

[0054] As a further preferred solution, the method for creating supersaturation in the salt-forming system in step 2) of the above preparation method includes one or more of the following: volatilizing the solvent, adding an anti-solvent, or cooling.

[0055] As a further preferred embodiment, the organic solvent in the above preparation method is selected from alcohols, chloroalkanes, ketones, ethers, cyclic ethers, esters, alkanes, cycloalkanes, benzenes, amides or sulfoxides, or mixtures thereof, or aqueous solutions thereof.

[0056] As a further preferred embodiment, the organic solvent is selected from methanol, ethanol, n-propanol, isopropanol, dichloromethane, heptane, acetonitrile, acetone, methyl ethyl ketone, toluene, 1,4-dioxane, tetrahydrofuran, 2-methyltetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, ethyl acetate, isopropyl acetate, methyl tert-butyl ether or 2-methoxyethyl ether, or a mixture thereof, or an aqueous solution thereof.

[0057] The fourth aspect of the present invention provides a crystalline form of the free acid of the compound of formula (I):

[0058] As a preferred embodiment, the crystalline form of the free acid of the compound of formula (I) is Form A, and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 5.56±0.2°, 6.55±0.2°, 7.24±0.2°, 9.83±0.2°, 11.13±0.2°, 13.14±0.2°, 15.81±0.2°, 18.01±0.2°, 19.75±0.2°, 20.73±0.2°, 22.38±0.2°, 23.10±0.2°, 23.37±0.2° and 26.28±0.2°.

[0059] As the most preferred embodiment, the X-ray powder diffraction (XRPD) of the crystalline form A includes substantially the same peaks as those at the diffraction angle (2θ) shown in FIG9 , and the X-ray powder diffraction data thereof are shown in the following table:

[0060] This crystalline form of the free acid compound of formula (I) is designated Form A.

[0061] As a preferred embodiment, the crystalline form of the free acid of the compound of formula (I) is Form B, and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 4.76±0.2°, 7.21±0.2°, 15.17±0.2°, 16.58±0.2°, 17.03±0.2°, 17.90±0.2°, 19.12±0.2°, 21.32±0.2°, 22.05±0.2°, 23.54±0.2°, 24.32±0.2°, 24.58±0.2°, 25.73±0.2°, 26.08±0.2° and 26.70±0.2°.

[0062] As the most preferred embodiment, the X-ray powder diffraction (XRPD) of the crystalline form B includes substantially the same peaks at the diffraction angle (2θ) as shown in FIG8 , and the X-ray powder diffraction data thereof are shown in the following table:

[0063] This crystalline form of the free acid compound of formula (I) is designated Form B.

[0064] The fourth aspect of the present invention provides a pharmaceutical composition comprising a clinically effective amount of a basic salt of a compound of formula (I), or a crystalline free acid of a compound of formula (I); and a pharmaceutically acceptable carrier.

[0065] In a fifth aspect, the present invention provides a basic salt of a compound of formula (I), or a crystalline free acid of a compound of formula (I), or a pharmaceutical composition comprising a clinically effective amount of a basic salt of a compound of formula (I) or a crystalline free acid of a compound of formula (I) in the preparation of a medicament for treating tumors, immune-related diseases and disorders or metabolic diseases mediated at least in part by CD73.

[0066] As a further preferred embodiment, the tumor is selected from prostate cancer, colon cancer, rectal cancer, pancreatic cancer, gastric cancer, endometrial cancer, cervical cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer, neck cancer, skin cancer, mesothelial lining cancer, white blood cell cancer, esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, lung cancer, adrenal cancer, thyroid cancer, kidney cancer, bone cancer, brain tumor, glioblastoma, mesothelioma, renal cell carcinoma, sarcoma, choriocarcinoma, epidermal basal cell carcinoma and testicular seminoma.

[0067] As a further preferred embodiment, the tumor is selected from skin cancer, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia cancer, brain tumor, ovarian cancer and sarcoma.

[0068] As a further preferred embodiment, the skin cancer is melanoma and basal cell carcinoma; the white blood cell cancer is lymphoma and leukemia; the lung cancer is small cell lung cancer and non-small cell carcinoma; and the sarcoma is Kaposi's sarcoma.

[0069] As a further preferred embodiment, the immune-related diseases and disorders are selected from rheumatoid arthritis, renal failure, lupus erythematosus, asthma, psoriasis, ulcerative colitis, pancreatitis, allergies, fibrosis, anemia, fibromyalgia, Alzheimer's disease, congestive heart failure, stroke, aortic valve stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infection, Crohn's disease, ulcerative colitis, allergic contact dermatitis and eczema, systemic sclerosis and multiple sclerosis.

[0070] In a sixth aspect, the present invention provides a basic salt of a compound of formula (I), or a crystalline free acid of a compound of formula (I), or a pharmaceutical composition comprising a clinically effective amount of a basic salt of a compound of formula (I) or a crystalline free acid of a compound of formula (I) for use as a drug for treating tumors, immune diseases and disorders or metabolic diseases mediated at least in part by CD73.

[0071] As a further preferred embodiment, the tumor is selected from prostate cancer, colon cancer, rectal cancer, pancreatic cancer, gastric cancer, endometrial cancer, cervical cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer, neck cancer, skin cancer, mesothelial lining cancer, white blood cell cancer, esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, small cell lung cancer, adrenal cancer, thyroid cancer, kidney cancer, bone cancer, brain tumor, glioblastoma, mesothelioma, renal cell carcinoma, sarcoma, choriocarcinoma, epidermal stromal tumor, ovarian cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, pelvic floor cancer, Basal cell carcinoma and testicular seminoma; the immune-related diseases and disorders are selected from rheumatoid arthritis, renal failure, lupus erythematosus, asthma, psoriasis, ulcerative colitis, pancreatitis, allergies, fibrosis, anemia, fibromyalgia, Alzheimer's disease, congestive heart failure, stroke, aortic valve stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infection, Crohn's disease, ulcerative colitis, allergic contact dermatitis and eczema, systemic sclerosis and multiple sclerosis.

[0072] The present invention also relates to a method for treating tumors, immune diseases and disorders and metabolic diseases mediated at least in part by CD73, comprising administering to a patient in need thereof a therapeutically effective amount of a basic salt of a compound of formula (I), or a crystalline form of a free acid of a compound of formula (I), or a pharmaceutical composition comprising a clinically effective amount of a basic salt of a compound of formula (I) or a crystalline form of a free acid of a compound of formula (I).

[0073] As a further preferred embodiment, the tumor is selected from prostate cancer, colon cancer, rectal cancer, pancreatic cancer, gastric cancer, endometrial cancer, cervical cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer, neck cancer, skin cancer, mesothelial lining cancer, white blood cell cancer, esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, small cell lung cancer, adrenal cancer, thyroid cancer, kidney cancer, bone cancer, brain tumor, glioblastoma, mesothelioma, renal cell carcinoma, sarcoma, choriocarcinoma, epidermal Basal cell carcinoma and testicular seminoma; the immune diseases and disorders are selected from rheumatoid arthritis, renal failure, lupus erythematosus, asthma, psoriasis, ulcerative colitis, pancreatitis, allergies, fibrosis, anemia, 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 eczema, systemic sclerosis and multiple sclerosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1 is an X-ray powder diffraction pattern of the free acid of the compound of formula (I). The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0075] Figure 2 shows the DSC spectrum of the free acid of the compound of formula (I). The abscissa represents temperature (°C) and the ordinate represents heat flow (w / g).

[0076] Figure 3 shows the mDSC spectrum of the free acid of the compound of formula (I). The abscissa represents temperature (°C), the first ordinate from the left represents reversing heat flow (w / g), the second ordinate from the left represents total heat flow (w / g), and the right ordinate represents irreversing heat flow (w / g).

[0077] Figure 4 is a TGA spectrum of the free acid of the compound of formula (I). The abscissa represents temperature (°C), and the ordinate represents weight (%).

[0078] Figure 5 Free acid of compound of formula (I) 1 H-NMR spectrum. The horizontal axis represents the compound displacement (ppm), and the vertical axis represents the signal intensity.

[0079] Figure 6 shows the X-ray powder diffraction pattern of the pentasodium salt of the compound of formula (I). The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0080] Figure 7 Pentasodium salt of compound of formula (I) 1 H-NMR spectrum. The horizontal axis represents the compound displacement (ppm), and the vertical axis represents the signal intensity.

[0081] Figure 8 is an X-ray powder diffraction pattern of the free acid crystalline form B of the compound of formula (I). The abscissa represents the 2θ value (degrees), and the ordinate represents the peak intensity.

[0082] Figure 9 shows an X-ray powder diffraction pattern of the free acid crystalline form A of the compound of formula (I). The abscissa represents the 2θ value (degrees), and the ordinate represents the peak intensity.

[0083] Figure 10 is an X-ray powder diffraction pattern of the monosodium salt of the amorphous form (I) compound. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0084] Figure 11 is an X-ray powder diffraction pattern of the trisodium salt of the amorphous form (I) compound. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0085] Figure 12 shows the X-ray powder diffraction pattern of the monopotassium salt of the amorphous form (I) compound. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0086] Figure 13 shows the X-ray powder diffraction pattern of the monocalcium salt of the amorphous form (I) compound. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0087] Figure 14 shows an X-ray powder diffraction pattern of the arginine salt of the amorphous form (I) compound. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0088] Figure 15 shows an X-ray powder diffraction pattern of the meglumine salt of the amorphous form (I) compound. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0089] Figure 16 shows an X-ray powder diffraction pattern of the diammonium salt of the amorphous form (I) compound. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0090] Figure 17 Amorphous Form (I) of the compound meglumine salt 1 H-NMR spectrum. The horizontal axis represents the compound displacement (ppm), and the vertical axis represents the signal intensity.

[0091] Figure 18 shows the DSC spectrum of the free acid crystalline form B of the compound of formula (I). The abscissa represents temperature (°C), and the ordinate represents heat flow (w / g).

[0092] Figure 19 is a TGA spectrum of the free acid crystalline form B of the compound of formula (I). The abscissa represents temperature (°C), and the ordinate represents weight (%).

[0093] Figure 20 Free acid form B of compound of formula (I) 1 H-NMR spectrum. The horizontal axis represents the compound displacement (ppm), and the vertical axis represents the signal intensity.

[0094] Figure 21 shows the mDSC spectrum of the trisodium salt of the compound of formula (I). The abscissa represents temperature (°C), the left ordinate represents reversing heat flow (w / g), and the right ordinate represents irreversing heat flow (w / g). The curves on the figure, from bottom to top, represent total heat flow (W / g), irreversing heat flow (W / g), and reversing heat flow (W / g), respectively.

[0095] Figure 22 is a TGA spectrum of the trisodium salt of the compound of formula (I). The abscissa represents temperature (°C), and the ordinate represents weight (%).

[0096] Figure 23: Trisodium salt of compound of formula (I) 1 H-NMR spectrum. The horizontal axis represents the compound displacement (ppm), and the vertical axis represents the signal intensity.

[0097] Figure 24 shows the mDSC spectrum of the monocalcium salt of the compound of formula (I). The abscissa represents temperature (°C), and the ordinate represents heat flow (w / g). The first ordinate from the left represents reversing heat flow (w / g), the second ordinate from the left represents total heat flow (w / g), and the right ordinate represents irreversing heat flow (w / g).

[0098] Figure 25 is a TGA spectrum of the monocalcium salt of the compound of formula (I). The abscissa represents temperature (°C), and the ordinate represents weight (%).

[0099] Figure 26: Monocalcium salt of compound of formula (I) 1 H-NMR spectrum. The horizontal axis represents the compound displacement (ppm), and the vertical axis represents the signal intensity.

[0100] FIG27 is an XRPD spectrum of the trisodium salt of the compound of formula (I) in Form A′, wherein the abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0101] FIG28 is a DSC spectrum of the trisodium salt of the compound of formula (I) in form A′, wherein the abscissa represents temperature (° C.) and the ordinate represents heat flow (w / g).

[0102] FIG29 is a TGA spectrum of the trisodium salt of the compound of formula (I) in form A′, wherein the abscissa represents temperature (° C.) and the ordinate represents weight (%).

[0103] Figure 30: Trisodium salt of compound of formula (I) in form A' 1 In the H-NMR spectrum, the horizontal axis represents the compound displacement (ppm) and the vertical axis represents the signal intensity.

[0104] FIG31 is an XRPD spectrum of the trisodium salt of the compound of formula (I) in form D′, wherein the abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0105] FIG32 is an XRPD spectrum of the trisodium salt of the compound of formula (I) in form E′, wherein the abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0106] FIG33 is a DSC spectrum of the trisodium salt of the compound of formula (I) in form E′, wherein the abscissa represents temperature (° C.) and the ordinate represents heat flow (w / g).

[0107] FIG34 is a TGA spectrum of the trisodium salt of the compound of formula (I) in form E′, wherein the abscissa represents temperature (° C.) and the ordinate represents weight (%).

[0108] Figure 35: Trisodium salt of compound of formula (I) in form E' 1 In the H-NMR spectrum, the horizontal axis represents the compound displacement (ppm) and the vertical axis represents the signal intensity.

[0109] FIG36 is an XRPD spectrum of the trisodium salt of the compound of formula (I) in form A1′, wherein the abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0110] FIG37 is a DSC spectrum of the trisodium salt of the compound of formula (I) in form A1′, wherein the abscissa represents temperature (° C.) and the ordinate represents heat flow (w / g).

[0111] FIG38 is a TGA spectrum of the trisodium salt of the compound of formula (I) in form A1′, wherein the abscissa represents temperature (° C.) and the ordinate represents weight (%).

[0112] Figure 39: Trisodium salt of compound of formula (I) in form A1' 1 In the H-NMR spectrum, the horizontal axis represents the compound displacement (ppm) and the vertical axis represents the signal intensity.

[0113] FIG40 is an XRPD spectrum of the trisodium salt of the compound of formula (I) in Form B′, wherein the abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0114] FIG41 is a DSC spectrum of the trisodium salt of the compound of formula (I) in Form B′, wherein the abscissa represents temperature (° C.) and the ordinate represents heat flow (w / g).

[0115] FIG42 is a TGA spectrum of the trisodium salt of the compound of formula (I) in Form B′, wherein the abscissa represents temperature (° C.) and the ordinate represents weight (%).

[0116] Figure 43: Trisodium salt of compound of formula (I) in form B' 1 In the H-NMR spectrum, the horizontal axis represents the compound displacement (ppm) and the vertical axis represents the signal intensity.

[0117] FIG44 is an XRPD spectrum of the trisodium salt of the compound of formula (I) in Form C′, wherein the abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0118] FIG45 is an XRPD spectrum of the trisodium salt of the compound of formula (I) in Form F', wherein the abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0119] FIG46 is a DSC spectrum of the trisodium salt of the compound of formula (I) in Form F', wherein the abscissa represents temperature (° C.) and the ordinate represents heat flow (w / g).

[0120] FIG47 is a TGA spectrum of the trisodium salt of the compound of formula (I) in Form F', wherein the abscissa represents temperature (° C.) and the ordinate represents weight (%).

[0121] FIG48 is a 1H-NMR spectrum of the trisodium salt of the compound of formula (I) in Form F', wherein the abscissa represents the compound displacement (ppm) and the ordinate represents the signal intensity.

[0122] Figure 49 shows the X-ray powder diffraction pattern of the free acid sample of the amorphous form (I) compound after DVS test. The abscissa represents the 2θ value (degrees) and the ordinate represents the peak intensity.

[0123] Figure 50 is an X-ray powder diffraction pattern of a sample of the free acid crystalline form B of the compound of formula (I) after DVS testing. The abscissa represents the 2θ value (degrees), and the ordinate represents the peak intensity.

[0124] Figure 51 shows the X-ray powder diffraction pattern of the trisodium salt of amorphous form (I) after DVS testing. The horizontal axis represents the 2θ value (degrees), and the vertical axis represents the peak intensity.

[0125] Figure 52 shows the X-ray powder diffraction pattern of the DVS-tested sample of the monocalcium salt of the amorphous form (I) compound. The horizontal axis represents the 2θ value (degrees) and the vertical axis represents the peak intensity.

[0126] Figure 53 is an XRPD overlay of the trisodium salt of the compound of formula (I), Form A1', before and after DVS testing. The abscissa represents the 2θ value (degrees), and the ordinate represents the peak intensity.

[0127] Figure 54 is an XRPD overlay of the trisodium salt of the compound of formula (I), Form E', before and after DVS testing. The abscissa represents the 2θ value (degrees), and the ordinate represents the peak intensity. DETAILED DESCRIPTION

[0128] The present invention studies the different aggregation forms of the compound of formula (I) and provides a basic salt and crystalline free acid of the compound of formula (I), which greatly improves the physicochemical properties of the free state of the compound of formula (I), such as chemical stability, solubility, and hygroscopicity. This enables the basic salt and crystalline free acid of the compound of formula (I) to meet the needs of clinical drug preparation development, has very important clinical application value, and can be widely used in the preparation of treatments for tumors, immune-related diseases, and metabolic diseases mediated at least in part by CD73, and is expected to accelerate the development of a new generation of CD73 inhibitor drugs. On this basis, the present invention is completed.

[0129] DETAILED DESCRIPTION: Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

[0130] In the present invention, the compound of formula (I) and the free acid of the compound of formula (I) have the same meaning and have the structure shown in the following formula (I):

[0131] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts, free acids, or prodrugs, along with other chemical components, and other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0132] "Clinically effective dose" refers to the dose or concentration that can cause the minimum pharmacological effect clinically, also known as the minimum effective dose or threshold dose.

[0133] The basic salts and free acids of the compound of formula (I) exhibit polymorphism or monomorphism. For example, the trisodium salt exhibits polymorphism. These "polymorphs" differ in their X-ray powder diffraction patterns, physicochemical and pharmacokinetic properties, and thermodynamic stability.

[0134] As used herein, "salt" refers to a compound prepared by reacting an organic acid or base drug with a pharmaceutically acceptable inorganic or organic acid or base.

[0135] (1) Methods and Materials

[0136] The reagents in the examples of the present invention are known and can be purchased commercially, or can be synthesized using or according to methods known in the art.

[0137] Unless otherwise specified, all reactions of the present invention are carried out under continuous magnetic stirring, with dry solvents and temperatures in degrees Celsius (° C.), with ambient conditions being 20-25° C. and 60-80% RH.

[0138] Unless otherwise specified, the various crystal forms referred to in the present invention may be an anhydrous crystal form or a crystal-containing crystal form. If it is a crystal-containing crystal form, preferably each crystal contains 1, 2, 3, 4 or 5 molecules of crystalline water, and more preferably each crystal contains 1 or 2 molecules of crystalline water.

[0139] (2)HPLC test method (HPLC)

[0140] The stability and solubility of the compound were tested using HPLC. The instrument model used was: Agilent 1260 infinityII. Wavelength: 220 nm.

[0141] Chromatographic column: XBridge BEH Shield RP18 (150 mm × 4.6 mm, 2.5 μm); detector: DAD; column temperature: 40°C; flow rate: 1.0 mL / min; diluent: acetonitrile:water (v:v = 1:1); mobile phase A: 10 mM ammonium acetate aqueous solution; mobile phase B: acetonitrile.

[0142] (3) X-ray powder diffraction test method (XRPD)

[0143] The crystal form of the sample was characterized by XRPD. The instrument model used was Bruker D8 Advance. The specific instrument parameters were as follows: X-ray path: reflection mode; detector: LYNXEYE_XE_T (1D mode); aperture angle: Max; radiation source: Cu / K-Alpha1 Main beam path slit: dual main motorized slit 10.0mm SollerMount soller axis 2.5°; secondary beam path slit: detector OpticsMount soller slit 2.5° dual secondary motorized slit 5.2mm; scanning mode: continuous scanning; scanning type: dual optical path mode; step length: 0.02°; time per step: 0.12s / step; scanning range: 3° to 40°; sample rotation speed: 15rpm; sample disk: single crystal silicon wafer, flat plate.

[0144] (4) Melting point and melting enthalpy test method (DSC)

[0145] The experimental method for characterizing samples using differential scanning calorimetry (DSC) involves taking a small amount of sample powder and placing it in a press-fit aluminum pan that comes with the instrument. After loading the sample, the pan is press-fitted with the pan, and then the pan is inserted into the instrument for testing. The DSC analysis used in this patent uses a TA Discovery 2500 or Q2000 instrument. Scan parameters are set as follows: a nitrogen atmosphere, a heating rate of 10°C / min, and a temperature range of 30 to 250°C.

[0146] (5) Modulated differential scanning calorimetry (mDSC)

[0147] Instrument: TA Discovery 2500; Sample: Tzero pan and Tzero sealed lid with 0.7 mm diameter hole; Pan temperature range: 0°C / 30°C to 250°C; Heating rate: 2°C / min; Nitrogen flow rate: 50 mL / min; Temperature adjustment range: + / -1°C / min; Sample size: approximately 0.5-2 mg.

[0148] (6) Thermogravimetric test method (TGA)

[0149] Thermogravimetric analysis (TGA) characterization involves taking a small amount of sample powder and placing it in an aluminum pan supplied with the instrument. The TGA analysis method used in this patent utilizes a TA Discovery 5500 or Q5000 instrument. Scan parameters are set to a nitrogen atmosphere and a heating rate of 10°C / min. The termination temperature is set at 300°C or when the remaining sample weight is less than 80% of the initial value.

[0150] (7) Residual solvent test method ( 1 H-NMR)

[0151] The residual solvent was determined by 1H-NMR analysis using a Bruker Avance-AV 400M instrument with a 5 mm PABBO BB / 19F-1H / D Z-GRD Z108618 / 0406 probe, 8 scans, a temperature of 297.6 K, and a relaxation delay of 1 second.

[0152] (8) Ion content test method (IC)

[0153] The sodium ion content was tested using ion chromatography (IC) using a Metrohm 940 professional IC instrument.

[0154] Detector: conductivity detector; eluent (anion): 3.2 mmol / L Na2CO3 + 1.0 mmol / L NaHCO3; eluent (cation): 2.5 mmol / L; MSA suppressor supplement: 0.5% H2SO4; chromatographic column: Anion A SUPP 5-150 or Cation Column C4-150; column temperature: 30°C; flow rate: 0.7 mL / min (anion) or 0.9 mL / min (cation); diluent: acetonitrile: water (v:v = 1:1).

[0155] (9) Water content test method (KF)

[0156] The moisture content of the samples was tested using the Karl Fischer (KF) coulometric method, and the instrument model used was Mettler Toledo Coulometric KF Titrator C30.

[0157] (10) Polarized light microscopy (PLM)

[0158] The samples were characterized using a polarizing microscope (PLM) using a BX53LED OLYMPUS instrument with a cross-polarized light source. The samples were dispersed in silicone oil.

[0159] (11) Dynamic moisture sorption (DVS)

[0160] The experimental method for characterizing samples using the dynamic moisture sorption method (DVS) is to take a small amount of sample powder and place it in a precision sample tray that comes with the instrument. After loading the sample, it is sent to the instrument for detection. The instrument model used for the dynamic moisture sorption method in this patent is DVS Intrinsic. The experimental parameters are set to use nitrogen as the carrier gas, set the constant temperature to 25°C, and use a gradient mode. The humidity changes are 40%-95%-0%-95%-40% RH. The humidity change of each gradient in the range of 0% to 90% is 10%, and the humidity change of the gradient in the range of 90% to 95% is 5%. The gradient endpoint is determined by the dm / dt method, and the gradient endpoint is determined by dm / dt being less than 0.002% and maintained for 10 minutes, or the longest maintenance time of each gradient is 240 minutes. After the test is completed, the sample is subjected to XRPD analysis to confirm whether the solid form has changed.

[0161] The present invention is further described in detail and completely by the accompanying drawings and the following examples, which are only used to illustrate specific embodiments of the present invention and should not be interpreted as limiting the scope of the present invention in any way.

[0162] Example 1

[0163] (Preparation of the free acid of the compound of formula (I) and its pentasodium salt)

[0164] (1) Preparation method of free acid of compound of formula (I)

[0165] Step 1: Synthesis of (2R,3R,4R,5R)-2-(acetoxymethyl)-5-(4,6-dichloro-1H-pyrazolo[3,4-b]pyridin-1-yl)tetrahydrofuran-3,4-diyl diacetate

[0166] 4,6-Dichloro-1H-pyrazolo[3,4-b]pyridine (3.1 g, 16.49 mmol) was placed in hexamethyldisilazane (30 mL), and ammonium sulfate (22 mg, 0.16 mmol) was added. The reaction mixture was heated to 140°C and stirred for 3 hours. The hexamethyldisilazane was removed by rotary evaporation under reduced pressure, and the residue was dissolved in acetonitrile (60 mL). β-D-ribofuranose 1,2,3,5-tetraacetate (5.8 g, 18.14 mmol) was added. The reaction mixture was cooled to 0°C in an ice bath. Trimethylsilyl trifluoromethanesulfonate (5.5 g, 24.7 mmol) was added dropwise with stirring and cooling in an ice bath. The reaction mixture was slowly warmed to room temperature and stirred overnight. The acetonitrile was removed by rotary evaporation under reduced pressure below 30°C. The residue was diluted with ethyl acetate and washed with saturated sodium bicarbonate. The layers were separated, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined and evaporated under reduced pressure. The residue was separated by column chromatography (ethyl acetate / petroleum ether = 15 / 85) to obtain (2R,3R,4R,5R)-2-(acetoxymethyl)-5-(4,6-dichloro-1H-pyrazolo[3,4-b]pyridin-1-yl)tetrahydrofuran-3,4-diyl diacetate (5.6 g, yield: 76%). MS m / z (ESI): 446.2, 448.2 [M+H] + .

[0167] Step 2: Synthesis of (2R,3R,4R,5R)-2-(acetoxymethyl)-5-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)tetrahydrofuran-3,4-diyl diacetate

[0168] (2R,3R,4R,5R)-2-(Acetoxymethyl)-5-(4,6-dichloro-1H-pyrazolo[3,4-b]pyridin-1-yl)tetrahydrofuran-3,4-diyl diacetate (0.40 g, 0.90 mmol) was dissolved in N-methylpyrrolidone (2.0 mL), and (R)-6-fluoro-2,3-dihydro-1H-inden-1-amine hydrochloride (0.25 g, 1.35 mmol) and N,N-diisopropylethylamine (0.46 g, 3.60 mmol) were added. The reaction was stirred in a sealed tube at 100 ° C for 44.3 hours. After completion of the reaction, water was added for dilution, and the mixture was extracted with ethyl acetate. The organic phase was washed with water and concentrated. The crude product was separated by column chromatography (EA / PE = 4 / 6) to give (2R,3R,4R,5R)-2-(acetoxymethyl)-5-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)tetrahydrofuran-3,4-diyl diacetate (0.44 g, yield 85%). MS m / z (ESI): 561.3 [M+H] + .

[0169] Step 3: Synthesis of (2R,3R,4S,5R)-2-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-5-(hydroxymethyl)tetrahydrofuran-3,4-diol

[0170] (2R,3R,4R,5R)-2-(Acetoxymethyl)-5-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)tetrahydrofuran-3,4-diyl diacetate (0.44 g, 0.78 mmol) was dissolved in methanol (3.0 mL) and potassium carbonate (0.54 g, 3.90 mmol) was added and the reaction was stirred at room temperature for 1 hour. After the reaction was completed, methanol was concentrated under reduced pressure, and the crude product was separated by column chromatography (DCM / MEOH = 14 / 1) and concentrated to give (2R,3R,4S,5R)-2-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (0.27 g, yield 76%). MS m / z (ESI): 435.2 [M+H] + .

[0171] Step 4: Preparation of (((((2R,3S,4R,5R)-5-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)hydroxyphosphino)methyl)phosphinocarboxylic acid

[0172] ((3aR,4R,6R,6aR)-6-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxazol-4-yl)methanol (0.27 g, 0.62 mmol) was dissolved in anhydrous trimethyl phosphate (3.0 mL), cooled to 0°C, and a solution of methylenebisphosphonic dichloride (0.62 g, 2.50 mmol) in trimethyl phosphate (2.0 mL) was slowly added dropwise. After the addition was complete, the reaction was continued at 0°C for 30 minutes. After completion, the reaction was quenched with water and stirred for 10 minutes. After that, the mixture was directly purified by reverse phase (H2O / ACN=4 / 1) to obtain the target compound (((((2R,3S,4R,5R)-5-(6-chloro-4-(((R)-6-fluoro-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)hydroxyphosphino)methyl)phosphinocarboxylic acid (89.4 mg, yield 24%). MS m / z (ESI): 593.0 [M+H] + .

[0173] 1 H NMR(400MHz,MeOH-d4)δ8.19(s,1H),7.28(dd,J=8.3,5.1Hz,1H),7.00(ddd,J=17.9,8.9,2.4Hz,2H),6.5 0(s,1H),6.33(d,J=3.7Hz,1H),5.32(t,J=7.3Hz,1H),4.71(dd,J=5.2,3.8Hz,1H),4.57(t,J=5.2Hz,1H) ,4.30(ddd,J=11.0,7.3,3.8Hz,1H),4.26–4.08(m,2H),3.05(ddd,J=16.1,8.7,3.9Hz,1H),2.93(dt,J=1 5.9,8.0Hz,1H),2.70(dtd,J=11.9,7.6,4.0Hz,2H),2.42(t,J=21.0Hz,2H),2.07(dq,J=12.9,8.0Hz,1H).

[0174] (2) Preparation method of pentasodium salt of compound of formula (I) (API:Na=1:5)

[0175] Disperse the free acid of the compound of formula (I) in water, add an excess of aqueous sodium hydroxide, and stir at room temperature until the solution becomes clear. Slowly add methanol until solid precipitates. Stir overnight at room temperature and collect the solid. Ion chromatography confirms that the molar ratio of API:Na is 1:5.

[0176] (3) Characterization of the free acid and pentasodium salt (API:Na=1:5) of the compound of formula (I)

[0177] By HPLC, XRPD, DSC, TGA, 1 The relevant properties of the compound of formula (I) and its sodium salt obtained above were measured by H-NMR, IC, and KF as follows. The specific test methods are detailed in the definition:

[0178] Example 2

[0179] (Screening of free acid crystal forms of compound of formula (I))

[0180] (1) 3-week suspension experiment at 25°C

[0181] Approximately 40 mg of the free acid of the compound of formula (I) was weighed, 0.1-0.5 mL of solvent was added, and the mixture was suspended at 25°C with magnetic stirring at 300 rpm for 3 weeks. The resulting suspension was centrifuged at 14,000 rpm using a 0.45 μm nylon membrane centrifuge tube. The resulting solid fraction was characterized by XRPD. The results are shown in the table below.

[0182] (2) 50℃ 3-week suspension experiment

[0183] Approximately 50 mg of the free acid compound of Formula (I) was weighed, and 0.1-0.5 mL of solvent was added. The suspension was stirred magnetically at 300 rpm and 50°C for 3 weeks. The resulting suspension was centrifuged at 14,000 rpm using a 0.45 μm nylon membrane centrifuge tube. The resulting solid fraction was characterized by XRPD. The results are shown in the table below.

[0184] (3) Antisolvent addition crystallization experiment

[0185] Approximately 40 mg of the free acid of the compound of formula (I) was weighed and thoroughly dissolved in 0.1-5 mL of a good solvent at ambient temperature (approximately 22-25°C). The resulting solution was passed through a 0.45 μm nylon membrane syringe filter to obtain a clear solution. One to four volumes of antisolvent were slowly added to the clear solution until a large amount of solid precipitated. The resulting sample was suspended at 25°C for one week. The resulting solid was characterized by polarizing light microscopy (PLM). The results are shown in the table below.

[0186] Example 3

[0187] (Salt type screening test)

[0188] The screening experiment was carried out on the selected counterions (basic cations) and screening solvents using the suspension method. About 50 mg of the free acid of the compound of formula (I) was weighed and placed in a 2 mL glass bottle with 1-3 equivalents of the counterion. An appropriate volume of screening solvent was added and the screening experiment was carried out using the suspension method. The obtained sample was suspended at 50°C for 2 hours, then naturally cooled to 25°C and suspended at 25°C for 1 week. The obtained suspension was centrifuged at 14,000 rpm through a 0.45 μm nylon filter membrane. The experimental results are detailed in the table below. The obtained solid was characterized by XRPD, and the obtained good solid was characterized by nuclear magnetic resonance ( 1 H-NMR) or ion chromatography (IC) to characterize the salt formation ratio.

[0189] (1) Experiment number RC0:

[0190] About 30 mg of pentasodium salt (API:Na=1:5) was weighed and suspended in 0.2 mL of ethanol at 25° C. for 3 days. The resulting solid was an amorphous trisodium salt (API:Na=1:3).

[0191] (2) Screening solvent - ethanol

[0192] (3) Screening solvent - acetone

[0193] (4) Screening solvent - dichloromethane

[0194] (5) Salt type characterization

[0195] The following 7 amorphous salt forms were obtained from the above screening experiment and were respectively 1 The results of H-NMR, IC and HPLC (note) characterization are shown in the table below.

[0196] Example 4

[0197] (Preparation of Free Acid Crystal Form B of the Compound of Formula (I))

[0198] The free acid crystalline form B of the compound of formula (I) was prepared and obtained according to the following steps.

[0199] 1) Weigh approximately 300 mg of the free acid of the compound of formula (I) and suspend it in 1 mL of isopropanol.

[0200] 2) The suspension was suspended at 300 rpm at 25°C for 4 days.

[0201] 3) Add 5 mg of Form B seed crystals (obtained in Example 2, Experiment EQ10) to the above suspension.

[0202] 4) 1 mL of isopropanol was added to the suspension, and the resulting suspension was suspended for another 2 weeks.

[0203] 5) The solid portion was collected by filtration and dried at 22-25°C / 10-20% RH for about 20 hours to obtain 215 mg of the free acid form B of the compound of formula (I) with a purity of 95.2% and a yield of 70%.

[0204] Characterization of the product revealed moderate crystallinity through XRPD patterns (Figure 8). DSC revealed no melting point after dehydration / desolvation at 31°C and 142°C (Figure 18). TGA revealed a weight loss of approximately 4.5% at 100°C and 2.2% between 100°C and 160°C (Figure 19). 1 H-NMR showed that it contained 0.3 equivalents of isopropanol, equivalent to about 2.8%, as shown in Figure 20.

[0205] Example 5

[0206] (Preparation of trisodium salt of amorphous form (I) compound)

[0207] (1) Preparation method 1:

[0208] The trisodium salt of the amorphous form of compound (I) was prepared and obtained according to the following steps:

[0209] 1) Weigh approximately 90 mg of the pentasodium salt of the compound of formula (I) (API:Na=1:5) and suspend in 0.6 mL of ethanol at 25°C for 2 days.

[0210] 2) The solid portion was collected by filtration and dried at 22-25° C. / 40-60% RH for about 2 hours.

[0211] 3) A total of 60 mg of the trisodium salt of amorphous form (I) of the compound was obtained as an off-white powder with a purity of 99.8% and a yield of 72%.

[0212] (2) Preparation method 2:

[0213] The trisodium salt of the amorphous form of compound (I) was prepared and obtained according to the following steps:

[0214] 1) Weigh 3.2 equivalents of sodium hydroxide (about 109 mg), add 3 mL of ethanol, and stir at 50°C for 30 min to obtain a clear solution.

[0215] 2) At 50° C., the free acid solution of the compound of formula (I) was slowly added to the sodium hydroxide solution, and the obtained sample was converted into a suspension.

[0216] 3) Add 5 mg of amorphous trisodium salt seeds (the product of the above method 1) to the above suspension.

[0217] 4) The suspension was stirred at 50°C for 2 hours, cooled to 25°C, and stirred at 25°C for 2 days.

[0218] 5) The solvent of the suspension was removed using a rotary evaporator, and the resulting solid was suspended in 3 mL of ethanol for another 3 days.

[0219] 6) The solid portion was collected by filtration and dried under vacuum at 35° C. for about 2 hours to obtain 502 mg of trisodium salt of amorphous form (I) as an off-white powder with a purity of 97.1% and a yield of 88%.

[0220] Through analysis of the above product, XRPD showed that it was nearly amorphous, as shown in Figure 11, and DSC showed that it had no obvious Tg, as shown in Figure 21. 1 H-NMR or IC showed that the stoichiometric ratio was 1:3, and TGA showed a weight loss of approximately 14.1% at 245°C, as shown in FIG22 . 1 H-NMR showed that it contained 0.7 equivalents of ethanol, equivalent to about 4.2%, as shown in Figure 23.

[0221] Note: The free acid of the compound of formula (I) in the above method 2 can be replaced by the free acid crystalline form B isopropanol-water solvate.

[0222] Example 6

[0223] (Scale-up preparation of amorphous monocalcium salt)

[0224] The amorphous form of compound (I) monocalcium salt was prepared and obtained according to the following steps.

[0225] 1) Weigh 500 mg of the free acid of the compound of formula (I), add 5 mL of ethanol, and stir at 50° C. for 0.5 hour to obtain a nearly clear solution.

[0226] 2) Weigh 1.05 equivalents of calcium hydroxide (about 62 mg) and add it to the above clear solution.

[0227] 3) About 5 mg of amorphous monocalcium salt seeds (RC5A product in Example 3.1) were added to the above suspension.

[0228] 4) The suspension was stirred at 50°C for 2 hours, cooled to 25°C, and stirred at 25°C for 2 days.

[0229] 5) The solid portion was collected by filtration and dried under vacuum at 30° C. for about 1 hour.

[0230] 6) A total of 511 mg of amorphous form (I) monocalcium salt was obtained with a purity of 97% and a yield of 95%.

[0231] The product was characterized by XRPD pattern showing that it was amorphous, as shown in Figure 13. DSC showed that it had no obvious Tg, as shown in Figure 24. 1 H-NMR or IC showed that the stoichiometric ratio was 1:1, and TGA showed a weight loss of approximately 8.6% at 150°C, as shown in FIG25 . 1 H-NMR showed that it contained 0.7 equivalents of ethanol, equivalent to about 4.6%, as shown in Figure 26.

[0232] Example 7

[0233] (Preparation of the trisodium salt crystal form of the compound of formula (I))

[0234] For the trisodium salt of the compound of formula (I), more than one hundred screening experiments were conducted under different screening conditions, and the following seven crystalline forms of the trisodium salt of the compound of formula (I) were obtained.

[0235] 7.1 Preparation of Trisodium Salt Form A' of the Compound of Formula (I)

[0236] (1) Preparation method 1:

[0237] 1) Weigh 50 mg of the trisodium salt of the compound of formula (I) and add 0.2 mL of tetrahydrofuran / water (v:v = 95:5).

[0238] 2) Suspend at 50°C with magnetic stirring at 400 rpm for 1 week.

[0239] 3) The resulting suspension was centrifuged and filtered at 14,000 rpm using a 0.45 μm nylon membrane centrifuge tube to obtain the trisodium salt form A' of the compound of formula (I).

[0240] (2) Preparation method 2:

[0241] 1) Weigh approximately 500 mg of the trisodium salt of amorphous form (I) of the compound and add it to an 8 mL glass bottle.

[0242] 2) Add 5 mL of a mixed solvent of tetrahydrofuran:water (v:v=95:5) and stir at 50° C. to obtain a suspension.

[0243] 3) 5 mg of Form A' seed crystals (product of Method 1) were added to the resulting suspension.

[0244] 4) The resulting suspension was suspended at 50°C with a magnetic stirrer at 400 rpm for 1 week.

[0245] 5) The obtained solid was collected by suction filtration and vacuum dried at 50° C. for about 2 hours to obtain 440 mg of the trisodium salt of the compound of formula (I) in the form A′ as an off-white powder with a yield of 80%.

[0246] The product was characterized by an XRPD pattern as shown in Figure 27 , a DSC pattern at 26.8°C, and no melting point after dehydration / desolvation as shown in Figure 28 . TGA showed a weight loss of approximately 14.5% at 200°C as shown in Figure 29 . 1 H-NMR showed that it contained 0.3 equivalents of tetrahydrofuran, equivalent to about 3.2%, as shown in Figure 30.

[0247] 7.2 Preparation of the trisodium salt of the compound of formula (I) in Form D'

[0248] 30 mg of the trisodium salt of the compound of formula (I) Form A' was weighed and placed at about 22-25°C / 60-80% RH for 12 hours. Form A' was completely transformed into Form D'. The XRPD pattern thereof is shown in FIG31 .

[0249] 7.3 Preparation of the trisodium salt of the compound of formula (I) in Form E'

[0250] 30 mg of the trisodium salt of the compound of Formula (I), Form A', was conditioned at approximately 22-25°C / 60-75% RH for 20 hours, followed by 22-25°C, 20-30% RH for 30 minutes to yield Form E' with a purity of 97.9%. Its XRPD characterization is shown in Figure 32. The DSC analysis revealed no melting point at 22.6°C after dehydration (see Figure 33). The TGA analysis showed a weight loss of approximately 10.6% at 200°C (see Figure 34). 1 H-NMR showed that it contained 0.1 equivalent of tetrahydrofuran, equivalent to about 1.1%, as shown in Figure 35. KF measured its water content to be 9.3%.

[0251] 7.4 Preparation of Trisodium Salt Form A1' of the Compound of Formula (I)

[0252] 200 mg of the trisodium salt of the compound of formula (I), Form A', was weighed and placed in an open container at 22-25°C / 75% RH for 2 days, followed by approximately 30 minutes at 22-25°C / 20-30% RH. This yielded Form A1' with low crystallinity and 97.8% purity. Its XRPD pattern is shown in Figure 36. The DSC analysis revealed no melting point at 23.9°C after dehydration (see Figure 37). The TGA analysis showed a weight loss of approximately 16.1% at 245°C (see Figure 38). 1H-NMR showed that it contained 0.1 equivalent of tetrahydrofuran, equivalent to about 1.1%, as shown in Figure 39. KF measured its water content to be 11.5%.

[0253] 7.5 Preparation of Trisodium Salt Form B' of the Compound of Formula (I)

[0254] (1) Preparation method 1:

[0255] 1) Weigh 50 mg of the trisodium salt of the compound of formula (I) and add 0.2 mL of methylpyrrolidone.

[0256] 2) Suspend at 25°C with magnetic stirring at 400 rpm for 2 weeks.

[0257] 3) The resulting suspension was centrifuged and filtered at 14,000 rpm using a 0.45 μm nylon membrane centrifuge tube to obtain the trisodium salt of the compound of formula (I) in Form B'.

[0258] (2) Preparation method 2:

[0259] 1) Weigh 50 mg of the trisodium salt of the compound of formula (I) and add 0.2 mL of methylpyrrolidone.

[0260] 2) Suspend at 50°C with magnetic stirring at 400 rpm for 1 week.

[0261] 3) The resulting suspension was centrifuged and filtered at 14,000 rpm using a 0.45 μm nylon membrane centrifuge tube to obtain the trisodium salt of the compound of formula (I) in Form B'.

[0262] The product was characterized by an XRPD pattern as shown in Figure 40, and DSC showed no melting point at 19.1°C after dehydration / desolvation, as shown in Figure 41. TGA showed a weight loss of approximately 20.7% at 245°C, as shown in Figure 42. 1 H-NMR showed that it contained 0.55 equivalents of methylpyrrolidone, equivalent to about 8.5%, as shown in Figure 43.

[0263] 7.6 Preparation of the trisodium salt of the compound of formula (I) in Form C'

[0264] 1) Weigh 50 mg of the trisodium salt of the compound of formula (I) and add 0.2 mL of 1,4-dioxane / water (V:V = 95:5).

[0265] 2) Suspend at 25°C with magnetic stirring at 400 rpm for 2 weeks.

[0266] 3) The resulting suspension was centrifuged and filtered at 14,000 rpm using a 0.45 μm nylon membrane centrifuge tube to obtain the trisodium salt of the compound of formula (I), Form C′. XRPD characterization is shown in FIG44 .

[0267] 7.7 Preparation of the trisodium salt of the compound of formula (I) in Form F'

[0268] 200 mg of the trisodium salt of the compound of Formula (I), Form C', was weighed and dried under vacuum at 50°C for 1 hour. Form C' transformed into Form F', as characterized by XRPD (Figure 45). DSC analysis at 30.0°C revealed no melting point after desolvation (Figure 46). TGA analysis showed a weight loss of approximately 15.3% at 230°C (Figure 47). 1 H-NMR showed that it contained 1.5 equivalents of 1,4-dioxane, equivalent to about 16.8%, as shown in Figure 48.

[0269] Example 8

[0270] (Solid physical and chemical stability)

[0271] This experiment examines the solid stability of the free acid of the amorphous form of the compound (I), the free acid crystal form B (isopropyl alcohol-water solvate) of the compound (I), the trisodium salt of the amorphous form of the compound (I), the monocalcium salt of the amorphous form of the compound (I), the trisodium salt crystal form A1' of the compound (I), and the trisodium salt crystal form E' of the compound (I) under the following experimental conditions.

[0272] 1) BS1: solid, 25°C / 92%RH, open conditions, 1 week.

[0273] 2) BS2: solid, 40°C / 75%RH, open conditions, 1 week.

[0274] 3) BS3: solid, 60°C, sealed container, 1 week.

[0275] 4) BS4: solid, light (visible light, 1.2 million lumens per hour).

[0276] 5) BS5: solid, 25°C / 92%RH, open conditions, 8 weeks.

[0277] 6) BS6: solid, 40°C / 75%RH, open conditions, 8 weeks.

[0278] 7) BS7: solid, 60°C, sealed container, 8 weeks.

[0279] The test results are as follows:

[0280] (1) Changes in chemical purity

[0281] (2) Changes in physical state

[0282] (3) Appearance color change

[0283] From the above experimental results, it can be seen that the purity of amorphous free acid and free acid crystal form B decreases significantly under the conditions of 40°C / 75% RH, 60°C and light, and they are chemically unstable.

[0284] The crystalline free acid form B is physically unstable and transforms into an amorphous form under the conditions of 25°C / 92% RH, 40°C / 75% RH, and 60°C and light, which is physically unstable.

[0285] Compared with the free acid crystalline form B, the amorphous monocalcium salt and the amorphous trisodium salt exhibit excellent chemical purity and stability. The chemical properties are stable under 25°C / 92%RH, 40°C / 75%RH, 60°C and light conditions.

[0286] The trisodium salt forms A1' and E' are chemically stable at 25°C / 92%RH, 40°C / 75%RH, and 60°C.

[0287] Compared with the amorphous trisodium salt, the chemical purity of the amorphous monocalcium salt is slightly reduced (about 1%) after light exposure, but the light degradation problem of the monocalcium salt can be effectively avoided under light-shielding conditions.

[0288] Compared with the amorphous trisodium salt, the crystalline trisodium salts A1' and E' transformed into amorphous form at 25°C / 92%RH and 40°C / 75%RH, deliquesced, and were accompanied by a slight color change, indicating physical instability.

[0289] Therefore, compared with the amorphous free acid, free acid crystalline form B and amorphous monocalcium salt, the amorphous trisodium salt has a significant advantage in chemical stability. Compared with the crystalline trisodium salts A1' and E', the amorphous trisodium salt has a significant advantage in physical stability.

[0290] Example 9

[0291] (Solubility test)

[0292] A certain amount of the free acid of the amorphous form of the compound (I); the free acid crystalline form B (isopropanol-water solvate) of the compound of formula (I); and the trisodium salt of the amorphous form of the compound (I) were accurately weighed and placed in 2 mL glass bottles (the masses of the weighed free acid and candidate salt were equivalent to 10 mg of anhydrous free acid, respectively). 1 mL of solvents of different pH values ​​(preparation methods are shown in the table below) were added to obtain clear solutions, and the pH values ​​of the clear solutions were measured using a pH meter.

[0293] Accurately weigh 2.3 mg of the amorphous form of the compound (I) monocalcium salt and place it in a 2 mL glass vial. The mass of the amorphous form of the compound (I) monocalcium salt is equivalent to 2 mg of the free acid anhydrous crystalline form of the compound (I). The resulting suspension was stirred at 400 rpm at 37°C for 24 hours. The sample was centrifuged at 14,000 rpm at 37°C for 5 minutes. The concentration of the supernatant was determined by HPLC, and the pH of the supernatant was determined by a pH meter. The residual solid fraction after the solubility experiment was characterized by XRPD. The test results are shown in the table below.

[0294] To 200 mg of the trisodium salt form A1' of the compound of formula (I) (equivalent to approximately 150 mg of the anhydrous free acid) was added a pH 7.4 phosphate buffer solution (50 mM) at ambient temperature (approximately 22-25°C), and the pH of the resulting solution was adjusted to 7.8. The solution was then stirred at 25°C for 3 days, with no apparent precipitation.

[0295] Amorphous Form (I) free acid, Form B free acid of the compound of Formula (I), and trisodium salt of Amorphous Form (I) all exhibited high solubility (>10 mg / mL) in these five aqueous buffers at different pH values. The extent of pH shift increased with increasing concentration in pH 4.5 acetate buffer, pH 6.8 phosphate buffer, and pH 7.4 phosphate buffer.

[0296] The trisodium salt form A1' of the compound of formula (I) has a very high solubility of about 150 mg / mL in pH 7.4 phosphate buffer, which is much higher than the solubility of the amorphous form of the monocalcium salt of the compound of formula (I).

[0297] The monocalcium salt of amorphous form (I) of the compound has a low solubility (~0.5 mg / mL) in all five aqueous buffers, and the solid obtained after the solubility experiment is still amorphous.

[0298] Example 10

[0299] (Hygroscopicity test)

[0300] The hygroscopicity of the amorphous free acid, free acid Form B, amorphous trisodium salt, and amorphous monocalcium salt was evaluated by dynamic moisture sorption (DVS) testing at 25 °C.

[0301] From the data analysis in the above table, we can see that:

[0302] 1) The free acid of the amorphous form (I) is highly hygroscopic, with a weight gain of approximately 14.7% when absorbing moisture between 0% and 80% RH at 25°C. After DVS testing, the obtained solid remained amorphous, as shown in Figure 49.

[0303] 2) The free acid Form B of the compound of formula (I) is highly hygroscopic, absorbing moisture and gaining approximately 13.1% weight between 0% and 80% relative humidity at 25°C. After DVS testing, the free acid Form B transformed into an amorphous form, as shown in Figure 50 .

[0304] 3) The trisodium salt of the amorphous form (I) is highly hygroscopic, with a weight gain of approximately 29.1% when absorbing moisture between 0% RH and 80% RH at 25°C. After DVS testing, the obtained solid was amorphous, as shown in FIG51 .

[0305] 4) The amorphous form (I) of the monocalcium salt of the compound is highly hygroscopic, with a weight gain of approximately 14.1% when absorbing moisture between 0% and 80% RH at 25°C. After DVS testing, the obtained solid was still amorphous, as shown in Figure 52.

[0306] 5) The trisodium salt of the compound of formula (I), Form A1′, is highly hygroscopic, with a weight gain of approximately 35.9 g at 25° C. between 0% RH and 80% RH. DVS testing revealed that the solid was amorphous, as shown in FIG53 .

[0307] 6) The trisodium salt of the compound of formula (I), Form E', is highly hygroscopic, absorbing approximately 33.0% of its weight between 0% RH and 80% RH at 25° C. DVS testing revealed that the solid was amorphous, as shown in FIG54 .

[0308] All documents mentioned herein are incorporated herein by reference, just as if each document were individually incorporated by reference. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the present invention.

Claims

1. A basic salt of a compound of formula (I), in, The basic salt is sodium salt, potassium salt, calcium salt, arginine salt, meglumine salt or ammonium salt.

2. A basic salt of a compound of formula (I) according to claim 1, characterized in that The basic salt is a sodium salt or a calcium salt.

3. The basic salt of the compound of formula (I) according to claim 1, characterized in that The molar ratio of the compound of formula (I) to the base in each molecule of the basic salt of the compound of formula (I) is 1:1 to 1:5; preferably, the molar ratio of the compound of formula (I) to the base is 1:1, 1:2, 1:3, 1:4 or 1:

5.

4. The basic salt of the compound of formula (I) according to claim 1, characterized in that The basic salt of the compound of formula (I) is the sodium salt of the compound of formula (I), and the molar ratio of the compound of formula (I) to sodium atoms in each molecule of the sodium salt of the compound of formula (I) is 1:1, 1:3 or 1:

5.

5. The basic salt of the compound of formula (I) according to claim 4, characterized in that The molar ratio of the compound of formula (I) to sodium atoms in each molecule of the sodium salt of the compound of formula (I) is 1:

3.

6. The basic salt of the compound of formula (I) according to claim 4, characterized in that The sodium salt of the compound of formula (I) is an amorphous or crystalline compound.

7. The basic salt of the compound of formula (I) according to claim 6, characterized in that The basic salt of the compound of formula (I) is the trisodium salt of the amorphous form of the compound (I).

8. The basic salt of the compound of formula (I) according to claim 6, characterized in that The basic salt of the compound of formula (I) is a crystalline trisodium salt of the compound of formula (I).

9. The basic salt of the compound of formula (I) according to claim 8, characterized in that The crystalline form of the trisodium salt of the compound of formula (I) is Form A', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.22±0.2°, 6.38±0.2°, 9.55±0.2°, 12.72±0.2°, 19.10±0.2°, 20.01±0.2°, 20.53±0.2°, 23.13±0.2°, 25.50±0.2°, 26.27±0.2°, 26.66±0.2°, 28.80±0.2°, 29.43±0.2°, 32.06±0.2° and 35.37±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form A′ comprises peaks at diffraction angles (2θ) substantially the same as those shown in FIG. 27 .

10. The basic salt of the compound of formula (I) according to claim 8, characterized in that The crystalline form of the trisodium salt of the compound of formula (I) is Form B', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.15±0.2°, 6.27±0.2°, 9.34±0.2°, 12.34±0.2°, 12.59±0.2°, 15.64±0.2°, 17.06±0.2°, 18.90±0.2°, 23.60±0.2° and 25.93±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form B' comprises peaks at diffraction angles (2θ) substantially the same as those shown in FIG. 40 .

11. The basic salt of the compound of formula (I) according to claim 8, characterized in that The crystalline form of the trisodium salt of the compound of formula (I) is Form C', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 11.4±0.2°, 17.09±0.2°, 17.68±0.2°, 19.01±0.2°, 19.59±0.2°, 19.97±0.2°, 20.81±0.2°, 21.65±0.2°, 22.39±0.2°, 22.81±0.2°, 24.80±0.2°, 27.06±0.2°, 27.73±0.2°, 30.53±0.2° and 32.62±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form C' comprises peaks at diffraction angles (2θ) substantially the same as those shown in FIG. 44 .

12. The basic salt of the compound of formula (I) according to claim 8, characterized in that The crystalline form of the trisodium salt of the compound of formula (I) is Form D', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 11.97±0.2°, 17.97±0.2°, 18.92±0.2°, 19.41±0.2°, 20.36±0.2°, 20.97±0.2°, 21.66±0.2°, 22.88±0.2°, 23.24±0.2°, 24.15±0.2°, 24.77±0.2°, 25.11±0.2°, 25.51±0.2°, 26.21±0.2° and 30.10±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form D' comprises peaks at diffraction angles (2θ) substantially the same as those shown in FIG. 31 .

13. The basic salt of the compound of formula (I) according to claim 8, characterized in that The crystalline form of the trisodium salt of the compound of formula (I) is form E', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.15±0.2°, 3.24±0.2°, 9.79±0.2°, 12.35±0.2°, 12.41±0.2°, 14.30±0.2°, 18.61±0.2°, 19.91±0.2°, 20.80±0.2°, 23.94±0.2°, 24.89±0.2°, 31.39±0.2° and 34.67±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form E' comprises peaks at diffraction angles (2θ) substantially the same as those shown in FIG. 32 .

14. The basic salt of the compound of formula (I) according to claim 8, characterized in that The crystalline form of the trisodium salt of the compound of formula (I) is Form F', and its X-ray powder diffraction pattern (XRPD) includes peaks at diffraction angles (2θ) of 3.12±0.2°, 5.81±0.2°, 6.27±0.2°, 9.43±0.2°, 18.94±0.2° and 25.58±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form F' comprises peaks at diffraction angles (2θ) substantially the same as those shown in FIG. 45 .

15. The basic salt of the compound of formula (I) according to claim 8, characterized in that The crystalline form of the trisodium salt of the compound of formula (I) is form A1', and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 3.19±0.2°, 9.55±0.2°, 12.71±0.2°, 13.52±0.2°, 14.43±0.2°, 16.37±0.2°, 19.07±0.2°, 19.99±0.2°, 20.48±0.2°, 24.02±0.2°, 25.06±0.2°, 25.43±0.2°, 26.21±0.2°, 26.60±0.2° and 29.40±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form A1' comprises peaks at diffraction angles (2θ) substantially the same as those shown in FIG. 36 .

16. The basic salt of the compound of formula (I) according to claim 1, characterized in that The basic salt of the compound of formula (I) is a calcium salt of the compound of formula (I), and the molar ratio of each molecule of the compound of formula (I) to the calcium atom in the calcium salt of the compound of formula (I) is 1:

1.

17. A method for preparing a basic salt of the compound of formula (I) according to claim 1, characterized in that: The steps include: 1) dissolving or dispersing the free acid of the compound of formula (I) in water or an organic solvent, and adding an alkaline solution to the above system to carry out a salt-forming reaction; or, adding the free acid of the compound of formula (I) to an alkaline solution to carry out a salt-forming reaction; 2) collecting the solid product basic salt of the compound of formula (I) precipitated in the above-mentioned salt-forming reaction process, or obtaining the solid product basic salt of the compound of formula (I) by creating supersaturation in the salt-forming system; The free acid of the compound of formula (I) is an anhydrate, a hydrate or a solvate; The basic salt of the compound of formula (I) is a sodium salt, potassium salt, calcium salt, arginine salt, meglumine salt or ammonium salt of the compound of formula (I); The alkaline solution is selected from an aqueous solution or an organic solvent solution of sodium hydroxide, potassium hydroxide, calcium hydroxide, arginine, meglumine or ammonia.

18. A method for preparing a basic salt of the compound of formula (I) according to claim 6, characterized in that: The steps include: 1) dissolving or dispersing pentasodium salt of the compound of formula (I) in water or an organic solvent; 2) stirring or suspending, and collecting the trisodium salt solid of the compound of formula (I) precipitated in the above process.

19. The preparation method according to claim 17, characterized in that: The method of creating supersaturation in the salt-forming system in step 2) includes one or more of the following: volatilizing the solvent, adding an anti-solvent or lowering the temperature.

20. The preparation method according to claim 17 or 18, characterized in that: The organic solvent is selected from alcohols, chloroalkanes, ketones, ethers, cyclic ethers, esters, alkanes, cycloalkanes, benzenes, amides or sulfoxides, or mixtures thereof, or aqueous solutions thereof; Preferably, the organic solvent is selected from methanol, ethanol, n-propanol, isopropanol, dichloromethane, heptane, acetonitrile, acetone, methyl ethyl ketone, toluene, 1,4-dioxane, tetrahydrofuran, 2-methyltetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, ethyl acetate, isopropyl acetate, methyl tert-butyl ether or 2-methoxyethyl ether, or a mixture thereof, or an aqueous solution thereof.

21. A crystalline form of a compound of formula (I) free acid:

22. The crystalline form of the free acid of the compound of formula (I) according to claim 21, characterized in that The free acid of the crystalline form of the compound of formula (I) is Form A, and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 5.56±0.2°, 6.55±0.2°, 7.24±0.2°, 9.83±0.2°, 11.13±0.2°, 13.14±0.2°, 15.81±0.2°, 18.01±0.2°, 19.75±0.2°, 20.73±0.2°, 22.38±0.2°, 23.10±0.2°, 23.37±0.2° and 26.28±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form A is substantially the same as the peak at the diffraction angle (2θ) shown in FIG. 9 .

23. The crystalline form of the free acid of the compound of formula (I) according to claim 21, characterized in that The free acid of the crystalline form of the compound of formula (I) is Form B, and its X-ray powder diffraction pattern (XRPD) includes five or more peaks at diffraction angles (2θ) of 4.76±0.2°, 7.21±0.2°, 15.17±0.2°, 16.58±0.2°, 17.03±0.2°, 17.90±0.2°, 19.12±0.2°, 21.32±0.2°, 22.05±0.2°, 23.54±0.2°, 24.32±0.2°, 24.58±0.2°, 25.73±0.2°, 26.08±0.2° and 26.70±0.2°; Preferably, the X-ray powder diffraction pattern of the crystalline form B is substantially the same as the peak at the diffraction angle (2θ) shown in FIG8 .

24. A pharmaceutical composition comprising: A clinically effective amount of a basic salt of a compound of formula (I) according to any one of claims 1 to 16, or a crystalline form of a free acid of a compound of formula (I) according to any one of claims 21 to 23; and a pharmaceutically acceptable carrier.

25. Use of a basic salt of a compound of formula (I) as described in any one of claims 1 to 16, or a crystalline free acid of a compound of formula (I) as described in any one of claims 21 to 23, or a pharmaceutical composition as described in claim 24 in the preparation of a medicament for treating tumors, immune-related diseases and disorders or metabolic diseases mediated at least in part by CD73.

26. The use according to claim 25, characterized in that The tumor is selected from the group consisting of prostate cancer, colon cancer, rectal cancer, pancreatic cancer, gastric cancer, endometrial cancer, cervical cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer, neck cancer, skin cancer, mesothelial lining cancer, white blood cell cancer, esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, lung cancer, adrenal cancer, thyroid cancer, kidney cancer, bone cancer, brain tumor, glioblastoma, mesothelioma, renal cell carcinoma, sarcoma, choriocarcinoma, epidermal basal cell carcinoma and testicular seminoma; Preferably, the tumor is selected from the group consisting of skin cancer, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia cancer, brain tumor, ovarian cancer and sarcoma.

27. The use according to claim 25, characterized in that The immune-related diseases and disorders are selected from rheumatoid arthritis, renal failure, lupus erythematosus, asthma, psoriasis, ulcerative colitis, pancreatitis, allergies, fibrosis, anemia 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 eczema, systemic sclerosis and multiple sclerosis.

28. A basic salt of a compound of formula (I) as described in any one of claims 1 to 16, or a crystalline free acid of a compound of formula (I) as described in any one of claims 21 to 23, or a pharmaceutical composition as described in claim 24, for use as a medicament for treating tumors, immune diseases and disorders or metabolic diseases mediated at least in part by CD73.

29. The pharmaceutical composition according to claim 28, wherein the tumor is selected from the group consisting of prostate cancer, colon cancer, rectal cancer, pancreatic cancer, gastric cancer, endometrial cancer, cervical cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer, neck cancer, skin cancer, mesothelial lining cancer, white blood cell cancer, esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, small cell lung cancer, adrenal cancer, thyroid cancer, kidney cancer, bone cancer, brain tumor, glioblastoma, mesothelioma, renal cell carcinoma, sarcoma, choriocarcinoma cancer, epidermal basal cell carcinoma and testicular seminoma; the immune-related diseases and disorders are selected from rheumatoid arthritis, renal failure, lupus erythematosus, asthma, psoriasis, ulcerative colitis, pancreatitis, allergies, fibrosis, anemia, fibromyalgia, Alzheimer's disease, congestive heart failure, stroke, aortic valve stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infection, Crohn's disease, ulcerative colitis, allergic contact dermatitis and eczema, systemic sclerosis and multiple sclerosis.

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