Salts of dioxanequinoline compounds, their crystalline forms, their preparation method, and their uses

Abstract

The present invention provides a salt of a dioxanequinoline compound, its crystalline form, its preparation method, and its use. Specifically, the present invention relates to N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide, its crystalline form, its preparation method, and its use in preparing a medicament as a tyrosine kinase (e.g., VEGFR-2, c-MET, etc.) inhibitor. [C1] TIFF2025541949000023.tif43156

Description

[Technical Field]

[0001] The present disclosure relates to salts of N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide, crystalline forms thereof, processes for their preparation, and their use in the manufacture of medicaments as inhibitors of tyrosine kinases (VEGFR-2, c-MET, c-KIT, PDGF-Ra, RET, AXL, NTRK, etc.). [Background technology]

[0002] Vascular endothelial growth factor receptors (VEGFRs) are members of the receptor tyrosine kinase family. VEGFRs bind to their ligand, vascular endothelial growth factor (VEGF), initiating a series of biochemical and physiological processes that ultimately promote neovascularization. Tumor vascularization and permeability are primarily regulated by VEGF, which acts through at least two distinct receptors (VEGFR-1 and VEGFR-2). Studies by Jakeman, Kolch, Connolly, and others have shown that VEGF is a key stimulator of normal and abnormal angiogenesis and vascular permeability (Jakeman et al., 1993, Endocrinology 133:848-859; Kolch et al., 1995, Breast Cancer Research and Treatment 36:139-155; Connolly et al., 1989, J. Biol. Chem. 264:20017-20024). Vascular endothelial growth factor induces an angiogenic sprouting phenotype by inducing endothelial cell proliferation, protease expression, and migration, followed by the formation of capillary vascular tissue. Therefore, VEGF antagonism, achieved by antibody-mediated VEGF chelation, may lead to tumor growth inhibition (Kim et al., 1993, Nature 362:841-844).

[0003] VEGFR-2 is primarily distributed in vascular endothelial cells and can bind to VEGF-A, VEGF-C, VEGF-D, and VEGF-E. VEGF stimulates endothelial cell proliferation and increases vascular permeability and angiogenesis primarily by binding to and activating VEGFR-2. Inhibition of VEGFR-2 activity can suppress tumor growth and metastasis through both direct and indirect pathways, providing ideal antitumor efficacy. Therefore, discovering small molecule inhibitors with high activity and selectivity for VEGFR-2 has become a promising strategy for tumor treatment.

[0004] The hepatocyte growth factor receptor (c-MET) is a tyrosine kinase receptor, and aberrant activation of c-MET plays an important role in the development and progression of various malignant tumors, including lung cancer. Hepatocyte growth factor (HGF) is a specific ligand for c-MET. Upon binding to HGF, c-MET exerts its biological effects through the HGF / c-MET signaling pathway, which can induce a range of biological effects, including cell proliferation, scattering, migration, organ morphogenesis, and angiogenesis. Abnormal activation of c-MET can manifest as receptor overexpression, gene mutation, amplification, ectopic expression, and rearrangement. These alterations can dysregulate downstream signaling pathways, such as the serine / threonine protein kinase (AKT), extracellular signal kinase (ERK), phosphatidylinositol-3-hydroxykinase, and retinoblastoma inhibitory protein (Rb) pathways, mediating processes such as tumor initiation, invasion, and metastasis, as well as angiogenesis and epithelial-mesenchymal transition. c-MET plays an important role in cell proliferation, metabolism, tumorigenesis, metastasis, and angiogenesis, making it an important target for antitumor therapy. Targeted therapy targeting c-MET has shown promise in the treatment of various malignant tumors, including lung cancer. During antitumor drug therapy, interactions between multiple signaling pathways affect the efficacy of antitumor drugs. For example, interactions between the HFG / c-MET signaling pathway and other pathways can affect the therapeutic efficacy of antitumor drugs and lead to drug resistance. Therefore, multikinase-targeted combination therapy has become a novel antitumor treatment.

[0005] WO2019154133 (Application No. PCT / CN2019 / 073260, Filing Date: January 25, 2019) discloses a compound that can effectively inhibit VEGFR-2 and c-MET tyrosine kinase. The compound can also effectively suppress the proliferation of tumor cells such as MHCC97H. The structure of this compound is shown in Formula (I). [ka]

[0006] However, the compound represented by formula (I) has extremely low solubility in water and insufficient solubility in simulated gastrointestinal fluid. This significantly affects the solubility of the drug in the digestive tract, ultimately affecting the drug's absorption in the human body and, ultimately, its efficacy. Therefore, there is a strong demand for improving the physicochemical properties of the compound represented by formula (I) from various aspects (such as solubility, chemical stability, pharmacokinetic properties, and bioavailability). Summary of the Invention [Means for solving the problem]

[0007] The present disclosure provides pharmaceutically acceptable salts of the compound of formula (I), its crystalline forms, and methods for their preparation and use in the manufacture of medicaments as inhibitors of tyrosine kinases (VEGFR-2, c-MET, c-KIT, PDGFRa, RET, AXL, NTRK, etc.). The various salts and crystalline forms of the present disclosure have good physicochemical properties and biochemical activities.

[0008] The present disclosure provides pharmaceutically acceptable salts of compounds of formula (I). [ka]

[0009] The pharmaceutically acceptable salts are selected from inorganic and organic salts, alternatively p-toluenesulfonates, mucates, and phosphates.

[0010] Specifically, a pharmaceutically acceptable salt of the present disclosure can be a salt of N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide and p-toluenesulfonic acid in a chemical ratio of 1:1 or 1:2.

[0011] The present disclosure provides a process for preparing a pharmaceutically acceptable salt of a compound of formula (I), comprising the step of subjecting N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide to a salt-forming reaction with an inorganic or organic acid, alternatively p-toluenesulfonic acid, mucic acid, or phosphoric acid.

[0012] Specifically, the salt-forming reaction is carried out in a solvent, and the solvent is selected from water, an alcohol solvent, a halogenated hydrocarbon solvent, a ketone solvent, an ether solvent, a nitrile solvent, an ester solvent, an amide solvent, an aliphatic hydrocarbon solvent, an alicyclic hydrocarbon solvent, an aromatic hydrocarbon solvent, a mixed solvent of an alcohol solvent and water, a mixed solvent of a ketone solvent and water, a mixed solvent of an alcohol solvent and an ether solvent, a mixed solvent of a halogenated hydrocarbon solvent and a nitrile solvent, a mixed solvent of an amide solvent and water, or a mixed solvent of a nitrile solvent and water; the ketone solvent is alternatively acetone, the alcohol solvent is alternatively methanol, ethanol, or isopropanol, the ether solvent is alternatively diethyl ether, methyl tert-butyl ether, tetrahydrofuran, 2-methyltetrahydrofuran, or dioxane, the halogenated hydrocarbon solvent is alternatively dichloromethane or chloroform, and the nitrile solvent is alternatively acetone. the ester solvent is alternatively ethyl acetate, isopropyl acetate, or butyl acetate; the amide solvent is alternatively N,N-dimethylformamide or N,N-dimethylacetamide; the aliphatic hydrocarbon solvent is alternatively n-heptane; the alicyclic hydrocarbon solvent is alternatively cyclohexane; the aromatic hydrocarbon solvent is alternatively toluene, xylene, or isopropylbenzene; the mixed solvent of the alcohol solvent and ether solvent is alternatively a mixed solvent of diethyl ether and methanol; the mixed solvent of the halogenated hydrocarbon solvent and nitrile solvent is alternatively a mixed solvent of dichloromethane and acetonitrile; the mixed solvent of the alcohol solvent and water is alternatively a mixed solvent of methanol and water or a mixed solvent of ethanol and water; the mixed solvent of the ketone solvent and water is alternatively a mixed solvent of acetone and water; and the mixed solvent of the amide solvent and water is alternatively a mixed solvent of N,N-dimethylformamide and water.

[0013] The present disclosure provides a method for preparing p-toluenesulfonate salt of compound of formula (I), comprising forming a salt from N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide and p-toluenesulfonic acid in a chemical ratio of 1:1 or 1:2. The present disclosure also provides a method for preparing p-toluenesulfonate salt crystalline Form A-II of compound of formula (I), comprising weighing out specific amounts of compound of formula (I) and p-toluenesulfonic acid in a molar ratio of 1:1, suspending the resulting mixture in a solvent and stirring for a predetermined period of time, and then drying the resulting mixture. More specifically, the solvent may be acetone, dichloromethane, or dioxane.

[0014] The present disclosure further provides a method for preparing a mucic acid salt of the compound of formula (I), comprising weighing a specific amount of the compound of formula (I) and a mucic acid in a molar ratio of 0.5:1, suspending the mixture in a solvent and stirring for a predetermined period of time, and then vacuum drying. More specifically, the solvent can be acetone.

[0015] The present disclosure further provides a method for preparing a phosphate salt of the compound of formula (I), comprising weighing out a specific amount of the compound of formula (I) and phosphoric acid in a molar ratio of 1:1, suspending and stirring in a solvent for a predetermined period of time, and then vacuum drying. More specifically, the solvent may be ethanol / water.

[0016] The present disclosure further provides a method for preparing a crystalline form of a pharmaceutically acceptable salt of the compound of Formula (I), comprising weighing out a specific molar ratio of the compound of Formula (I) and an inorganic or organic acid, alternatively p-toluenesulfonic acid, mucic acid, or phosphoric acid, suspending the resulting mixture in a solvent and stirring for a specific period of time, and then drying the resulting mixture. The solvent is as described above. The method for preparing the crystalline form of the present disclosure may also include methods such as anti-solvent addition, slow evaporation, slow cooling, suspension stirring at room temperature or elevated temperatures, cyclic heating and cooling, gas-solid permeation, gas-liquid diffusion, and polymer derivatization.

[0017] The present disclosure provides a mucilage salt crystalline form AI of compound of formula (I), N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide, having a powder X-ray diffraction pattern with characteristic peaks at 2θ angles of 3.95, 5.54, 7.46, 10.87, and 16.58, with the error margin of error for each characteristic peak being ±0.2;

[0018] Alternatively, characteristic peaks are present at 2θ angles of 3.47, 3.95, 5.54, 7.46, 8.46, 10.48, 10.87, 14.90, 15.28, 16.16, and 16.58, and the error margin of the 2θ angle of each characteristic peak is ±0.2;

[0019] Further alternatively, there are characteristic peaks at 2θ angles of 3.47, 3.95, 5.54, 7.46, 8.46, 10.48, 10.87, 11.72, 14.08, 14.90, 15.28, 16.16, 16.58, 17.90, 20.29, 21.65, 22.35, 23.37, 24.53, 25.45, 27.35, and 32.58, and the error margin for 2θ angle of each characteristic peak is ±0.2.

[0020] The present disclosure provides p-toluenesulfonate crystalline Form A-II of compound of Formula (I), N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide, having an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 5.30, 6.54, 7.97, 8.78, 13.01, 15.86, 19.70, 19.94, and 20.62, with the error margin of 2θ angles for each characteristic peak being ±0.2;

[0021] Alternatively, characteristic peaks are present at 2θ angles of 5.30, 6.54, 7.97, 8.78, 10.52, 11.63, 11.81, 13.01, 15.86, 17.54, 19.34, 19.70, 19.94, 20.62, 22.50, and 25.95, and the error range of the 2θ angle of each characteristic peak is ±0.2;

[0022] Alternatively, the 2θ angles are 5.30, 6.54, 7.97, 8.78, 10.52, 11.63, 11.81, 13.01, 13.54, 14.33, 14.52, 15.06, 15.86, 16.46, 17.54, 18.02, 18.25, 18.75, 19.34, 19.70, 19.94, 20.02, 20.25, 20.34, 20.40, 20.54, 20.60, 20.70, 20.80, 20.9 ... Characteristic peaks are present at 0.62, 21.06, 21.46, 22.50, 23.67, 24.02, 24.35, 25.12, 25.95, 26.45, 27.20, 27.87, 29.20, 30.15, 30.87, and 31.83, and the error range of the 2θ angle of each characteristic peak is ±0.2.

[0023] The present disclosure provides di-p-toluenesulfonate crystalline Form A-IV of compound of Formula (I), N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide, having an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 6.36, 13.60, and 19.20, with the error margin of 2θ angles for each characteristic peak being ±0.2;

[0024] Alternatively, characteristic peaks are present at 2θ angles of 6.36, 9.87, 11.50, 13.60, 16.00, 19.20, 20.26, 20.86, and 21.36, and the error margin of the 2θ angle of each characteristic peak is ±0.2;

[0025] Further alternatively, there are characteristic peaks at 2θ angles of 6.36, 9.45, 9.87, 11.50, 12.88, 13.60, 14.40, 16.00, 19.20, 20.26, 20.86, 21.36, 22.59, 23.76, and 26.73, and the error margin for the 2θ angle of each characteristic peak is ±0.2.

[0026] The present disclosure provides a crystalline form AV of the phosphate salt of the compound of formula (I), N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide, having a powder X-ray diffraction pattern in which characteristic peaks are present at 2θ angles of 5.68, 7.56, and 16.93, with the error margin of 2θ angles for each characteristic peak being ±0.2;

[0027] Alternatively, characteristic peaks are present at 2θ angles of 4.03, 5.68, 6.84, 7.56, 9.36, 11.30, 11.94, 16.93, 19.76, and 23.01, and the error margin of the 2θ angle of each characteristic peak is ±0.2;

[0028] Further alternatively, there are characteristic peaks at 2θ angles of 4.03, 5.68, 6.84, 7.56, 9.36, 11.30, 11.94, 12.56, 13.69, 16.17, 16.93, 19.76, 22.02, 23.01, 23.98, and 25.19, and the error margin for the 2θ angle of each characteristic peak is ±0.2.

[0029] Another aspect of the present application provides pharmaceutical compositions comprising a pharmaceutically acceptable salt of a compound of formula (I) described herein, alternatively the p-toluenesulfonate, mucinate, or phosphate salt, or any one of mucinate salt crystalline form AI, p-toluenesulfonate salt crystalline form A-II, di-p-toluenesulfonate salt crystalline form A-IV, or phosphate salt crystalline form AV, and one or more pharmaceutically acceptable carriers or excipients.

[0030] The pharmaceutical compositions of the present application may also contain one or more additional therapeutic agents.

[0031] The present application also relates to the use of a pharmaceutically acceptable salt, crystalline form, or pharmaceutical composition of a compound of formula (I) in the manufacture of a medicament for treating a disease associated with the tyrosine kinases VEGFR-2, c-MET, c-KIT, PDGFRa, RET, AXL, or NTRK (NTRK includes TRK-A, TRK-B, TRK-C, and other TRK family kinases).

[0032] The disease may alternatively be cancer or an autoimmune disease, in particular ocular fundus diseases, xerophthalmia, psoriasis, vitiligo, dermatitis, alopecia areata, rheumatoid arthritis, colitis, multiple sclerosis, systemic lupus erythematosus, Crohn's disease, atherosclerosis, pulmonary fibrosis, liver fibrosis, myelofibrosis, non-small cell lung cancer, small cell lung cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, ovarian cancer, cervical cancer, colorectal cancer, melanoma, endometrial cancer, prostate cancer, bladder cancer, leukemia, gastric cancer, liver cancer, gastrointestinal stromal tumors, thyroid cancer, chronic granulocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, nasopharyngeal carcinoma, esophageal cancer, brain tumors, B-cell and T-cell lymphomas, lymphoma, multiple myeloma, bile duct carcinosarcoma or cholangiocarcinoma.

[0033] The present disclosure also relates to a method for treating a disease or condition mediated by kinases such as VEGFR-2, c-MET, c-KIT, PDGFRa, RET, AXL, NTRK, etc., comprising administering to a patient (human or other mammal, particularly human) in need thereof a therapeutically effective amount of a pharmaceutically acceptable salt, crystalline form, or pharmaceutical composition of a compound of Formula (I) described herein. The disease or condition mediated by kinases such as VEGFR-2, c-MET, c-KIT, PDGFRa, RET, AXL, NTRK, etc., includes those described above. [Brief explanation of the drawings]

[0034] The above and other objects, features, and advantages of the present disclosure will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings.

[0035] [Figure 1] FIG. 1 is an XRPD diagram of the mucilage salt crystalline form AI of the compound of formula (I).

[0036] [Figure 2] 1 is a TGA graph of the mucilage salt crystalline form AI of the compound of formula (I).

[0037] [Figure 3] 1 is a DSC graph of the mucilage salt crystalline form AI of the compound of formula (I).

[0038] [Figure 4] FIG. 1 is an XRPD diagram of crystalline form A-II of the p-toluenesulfonic acid salt of compound of formula (I).

[0039] [Figure 5] 1 is a TGA graph of crystalline form A-II of the p-toluenesulfonic acid salt of compound of formula (I).

[0040] [Figure 6] 1 is a DSC graph of crystalline form A-II of the p-toluenesulfonate salt of compound of formula (I).

[0041] [Figure 7] FIG. 1 is an XRPD diagram of crystalline form B-III of the p-toluenesulfonic acid salt of compound of formula (I).

[0042] [Figure 8] 1 is a TGA graph of crystalline form B-III of the p-toluenesulfonic acid salt of compound of formula (I).

[0043] [Figure 9] 1 is a DSC graph of crystalline form B-III of the p-toluenesulfonate salt of compound of formula (I).

[0044] [Figure 10] FIG. 1 is an XRPD diagram of crystalline form A-IV of the di-p-toluenesulfonate salt of the compound of formula (I).

[0045] [Figure 11] 1 is a TGA graph of crystalline form A-IV of the di-p-toluenesulfonate salt of compound of formula (I).

[0046] [Figure 12] 1 is a DSC graph of crystalline form A-IV of the di-p-toluenesulfonate salt of the compound of formula (I).

[0047] [Figure 13] FIG. 1 is an XRPD diagram of crystalline form AV of the phosphate salt of the compound of formula (I).

[0048] [Figure 14] 1 is a TGA graph of crystalline form AV of the phosphate salt of the compound of formula (I).

[0049] [Figure 15] 1 is a DSC graph of crystalline form AV of the phosphate salt of the compound of formula (I).

[0050] [Figure 16]1 is a graph showing the changes in tumor volume in the group administered with the compound of formula (I) in free form and in the negative control group.

[0051] [Figure 17] 1 is a graph showing changes in tumor volume in the group administered with the myxate crystalline form AI of the compound of formula (I) in Example 1 and the negative control group.

[0052] [Figure 18] 1 is a graph showing changes in tumor volume in the group administered with crystalline form A-II of p-toluenesulfonate of compound of formula (I) in Example 2 and the negative control group.

[0053] [Figure 19] 1 is a graph showing changes in tumor volume in the group administered with crystalline form AV of the phosphate salt of the compound of formula (I) in Example 5 and the negative control group. DETAILED DESCRIPTION OF THE INVENTION

[0054] Unless otherwise stated, the following terms used in this application (including the specification and claims) have the definitions set forth below. In this application, the use of "or" or "and" means "and / or" unless stated otherwise. In addition, the use of the term "comprises" and other forms such as "containing" and "having" is non-exclusive. Subheadings used herein are for organizational purposes only and should not be construed as limiting the topics described.

[0055] Unless otherwise specified, the term "C1-C6 alkyl" refers to an alkyl group containing 1 to 6 carbon atoms. Similarly, C1-C3 alkyl refers to an alkyl group containing 1 to 3 carbon atoms. For example, C1-C6 alkyl includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, and 2-methylpentyl.

[0056] The term "ether solvent" refers to a linear or cyclic compound containing an ether bond -O- and containing 1 to 10 carbon atoms. Specific examples include, but are not limited to, diethyl ether, dioxane, tetrahydrofuran, 2-methyltetrahydrofuran, propylene glycol methyl ether, or methyl tert-butyl ether.

[0057] The term "alcohol solvent" refers to a group derived by replacing one or more hydrogen atoms on a C1-C6 alkyl group with one or more hydroxyl groups, and examples include, but are not limited to, methanol, ethanol, isopropanol, n-propanol, isopentanol, or trifluoroethanol.

[0058] The term "ester solvent" refers to a conjugate of a lower organic acid containing 1 to 4 carbon atoms and a lower alcohol containing 1 to 6 carbon atoms. Specific examples include, but are not limited to, ethyl acetate, isopropyl acetate, or butyl acetate.

[0059] The term "ketone solvent" refers to a compound in which a carbon group (-CO-) is bonded to two hydrocarbon groups. Ketones are classified into aliphatic ketones, alicyclic ketones, aromatic ketones, saturated ketones, and unsaturated ketones depending on the hydrocarbon groups in the molecule. Specific examples include, but are not limited to, acetone, acetophenone, methyl isobutyl ketone, or methylpyrrolidone.

[0060] The term "nitrile solvent" refers to a group derived by replacing one or more hydrogen atoms on a C1-C6 alkyl group with one or more cyano groups, and specific examples include, but are not limited to, acetonitrile or propionitrile.

[0061] The term "aliphatic hydrocarbon solvent" refers to a chain hydrocarbon (e.g., a saturated aliphatic hydrocarbon) having 1 to 10 carbon atoms, including alkane solvents, specific examples of which include, but are not limited to, n-butane, n-pentane, n-hexane, or n-heptane.

[0062] The term "alicyclic hydrocarbon solvent" refers to a hydrocarbon compound having a cyclic carbon skeleton and 1 to 8 ring atoms, specific examples of which include, but are not limited to, cyclopentane or cyclohexane.

[0063] The term "amide solvent" refers to a compound containing a carbonylamino group (-CONH-) and having 1 to 10 carbon atoms, and specific examples include, but are not limited to, N,N-dimethylformamide or N,N-dimethylacetamide.

[0064] The term "aromatic hydrocarbon solvent" is a general term for carbocyclic compounds and their derivatives that have a closed cyclic conjugated system within the molecule, and the number of π electrons follows Huckel's rule. Specific examples include, but are not limited to, toluene, isopropylbenzene, or xylene.

[0065] The term "halogenated hydrocarbon solvent" refers to a group derived by replacing one or more hydrogen atoms on a C1-C6 alkyl group with one or more halogen atoms, including fluorine, chlorine, bromine, iodine, etc. Specific examples of "halogenated hydrocarbon solvents" include, but are not limited to, methyl chloride, dichloromethane, chloroform, or carbon tetrachloride.

[0066] The term "mixed solvent" refers to a solvent prepared by mixing one or more different organic solvents in a specific ratio, or a solvent prepared by mixing an organic solvent and water in a specific ratio. The mixed solvent may alternatively be one or more of a mixture of alcohol solvents, a mixture of alcohol solvents and ether solvents, a mixture of alcohol solvents and aliphatic hydrocarbon solvents, a mixture of ether solvents and aliphatic hydrocarbon solvents, a mixture of alcohol solvents and water, a mixture of ketone solvents and water, a mixture of halogenated hydrocarbon solvents and nitrile solvents, a mixture of amide solvents and water, or a mixture of ether solvents and water; the alcohol solvents, ether solvents, aliphatic hydrocarbon solvents, halogenated hydrocarbon solvents, amide solvents, and nitrile solvents are as defined above.

[0067] As used herein, the term "pharmaceutically acceptable salt" refers to organic and inorganic salts of compounds of the present disclosure that are suitable for use in humans and lower animals, without undue toxicity, irritation, allergic reaction, etc., and possess a reasonable benefit / risk ratio. Pharmaceutically acceptable salts of amines, carboxylic acids, phosphonates, and other types of compounds are well known in the art. Salts can be formed by reacting a compound of the present disclosure with a suitable free base or acid, including, but not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, perchloric acid, etc., or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, etc. Alternatively, salts can be obtained by methods well known in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, besylate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, glyconate, hemisulfate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, mesylate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, per-3-phenylpropionate, phosphate, picrate, propionate, stearate, sulfate, thiocyanate, p-toluenesulfonate, undecanoate, and the like. Representative alkali or alkaline earth metal salts include salts of sodium, lithium, potassium, calcium, magnesium, etc. Other pharmaceutically acceptable salts include suitable non-toxic salts of ammonium, quaternary ammonium, and amine cations formed from halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0068] The pharmaceutical compositions of the present disclosure comprise a pharmaceutically acceptable salt of a compound of Formula (I) as described herein or a crystalline form thereof, and an additional active agent such as a kinase inhibitor (small molecule, polypeptide, antibody, etc.), an immunosuppressant, an anti-cancer agent, an antiviral agent, an anti-inflammatory agent, an antifungal agent, an antibiotic, or an anti-angiogenic compound; and an optional pharmaceutically acceptable carrier, adjuvant, or excipient.

[0069] A pharmaceutically acceptable salt of a compound of formula (I) of the present disclosure, or a crystalline form thereof, may be used alone or in combination with one or more other drugs. When administered in combination, the therapeutic agents may be formulated for simultaneous administration or sequential administration at different times, or may be administered as a single composition. "Combination therapy" refers to the use of a compound of the present disclosure with another drug, either simultaneously or sequentially, with the goal of achieving optimal efficacy of the drugs. Simultaneous administration includes both simultaneous delivery dosage forms and separate dosage forms for each compound. Thus, the administration of a compound of the present disclosure can be combined with other therapies known in the art, such as radiation therapy, cytostatic agents, cytotoxic agents, or other anti-cancer agents used in the treatment of cancer, to improve cancer symptoms. The present disclosure does not limit the order of administration. A compound of the present disclosure may be administered before, simultaneously with, or after the other anti-cancer or cytotoxic agent.

[0070] To prepare the pharmaceutical composition of the present disclosure, a pharmaceutically acceptable salt of the compound of formula (I) or its crystalline form as an active ingredient can be intimately mixed with a pharmaceutical carrier. This can be carried out according to conventional pharmaceutical formulation techniques. The carrier can be used in a wide variety of forms depending on the formulation designed for various modes of administration (e.g., oral or parenteral administration). Suitable pharmaceutically acceptable carriers are well known in the art. Descriptions of some of these pharmaceutically acceptable carriers can be found in the Handbook of Pharmaceutical Excipients, jointly published by the American Pharmaceutical Association and the British Pharmaceutical Society.

[0071] The pharmaceutical compositions of the present disclosure may be in the form of, for example, tablets, capsules, pills, powders, sustained-release forms, solutions, or suspensions suitable for oral administration; clear solutions, suspensions, emulsions, or other forms for parenteral injection; ointments, creams, or other forms for topical application; or suppositories for rectal administration. The pharmaceutical component may also be provided in a unit dosage form for single administration of a precise dosage. The pharmaceutical component may include a conventional pharmaceutical carrier or excipient and a pharmaceutically acceptable salt or crystalline form thereof of a compound as an active ingredient prepared according to the present disclosure, and may further include other medicinal or pharmaceutical preparations, carriers, adjuvants, and the like.

[0072] Therapeutic compounds can also be administered to mammals other than humans. The dosage of a drug to a mammal depends on the species of the animal and its disease or disorder state. Therapeutic compounds can be administered to animals in the form of a capsule, bolus, tablet, or liquid. Therapeutic compounds can also be introduced into animals by injection or infusion. These dosage forms are prepared by traditional methods that meet veterinary standards of care. Alternatively, pharmaceutical compound drugs can be mixed with animal feed and fed to animals, so that concentrated feed additives or premixes can be prepared by mixing with regular animal feed. [Example]

[0073] The present disclosure will be described in more detail below with specific examples to clarify the objectives, technical solutions, and advantages of the present disclosure. It should be understood that the specific examples described herein are used only to illustrate the present disclosure and are not intended to limit the present invention. If specific techniques or conditions are not specified in the examples, the techniques or conditions described in the technical literature or product specifications shall be followed. If the manufacturer of the reagents or equipment used is not specified, the reagents or equipment are all commercially available conventional products. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items. The following examples are provided to better illustrate the present invention, and all temperatures refer to °C unless otherwise specified. The names of some compounds in the present disclosure have been generated and translated by Chemdraw. The present disclosure also provides methods for preparing corresponding pharmaceutically acceptable salts and crystalline forms. Various methods can be used to prepare the salts and crystalline forms described herein, including the following method. Alternatively, other methods known in the chemical arts can be used. Those skilled in the art will appreciate modifications to these methods. Alternative methods include, but are not limited to, the following:

[0074] Assay equipment and test conditions

[0075] Powder X-ray diffraction (XRPD): Measurement was performed using a PANalytacal X'Pert3 powder X-ray diffraction analyzer. Specific collected information is shown in Table 1 below.

[0076] [Table 1]

[0077] TGA Thermogravimetric Analysis: Tests were performed using a TA Q5000 / 5500 thermogravimetric analyzer, with a heating rate of 10°C / min and a temperature range from room temperature to the set endpoint temperature (see corresponding graph for details).

[0078] DSC Differential Scanning Calorimetry: Tests were performed using a TA Q200 / Q2000 / 2500 differential scanning calorimeter, with a heating rate of 10°C / min and a temperature range from 25°C to the set endpoint temperature (see corresponding graph for details).

[0079] Dynamic moisture sorption (DVS): Curves were collected on a DVS Intrinsic from Surface Measurement Systems (SMS) at 25°C, with a humidity range of 0% RH to 95% RH in 10% steps (with a final step of 5%) and a dm / dt of 0.002% / min.

[0080] High-performance liquid chromatography / ion chromatography (HPLC / IC): HPLC was performed using an Agilent 1260 / 1100 high-performance liquid chromatograph (Agilent Eclipse Plus C18, 100 × 4.6 mm, 3.5 μm, chromatography column). IC was performed using a ThermoFisher ICS-1100 ion chromatograph (IonPac AS18 analytical column, 250 × 4 mm, chromatography column).

[0081] Preparation of biological solvents

[0082] 1. Preparation of simulated gastric fluid (SGF): 0.1 g of NaCl and 0.05 g of Trinaton X-100 were weighed into a 50 mL volumetric flask and completely dissolved in purified water. 67.5 μL of concentrated hydrochloric acid (12 M) was added, and the mixture was adjusted to a pH of 1.8 with 1 M hydrochloric acid or 1 M NaOH aqueous solution. The mixture was then brought to volume with purified water.

[0083] 2. Preparation of fasted-state simulated intestinal fluid (FaSSIF): 0.17 g of anhydrous NaH2PO4, 0.021 g of NaOH, and 0.31 g of NaCl were weighed into a 50 mL volumetric flask. Approximately 48 mL of purified water was added to completely dissolve the mixture, and the solution was adjusted to a pH of 6.5 with 1 M hydrochloric acid or 1 M NaOH aqueous solution. Purified water was added to the volume. 0.11 g of SIF powder was added and completely dissolved.

[0084] 3. Preparation of fed-state simulated intestinal fluid (FeSSIF): 0.41 mL of glacial acetic acid, 0.20 g of NaOH, and 0.59 g of NaCl were weighed into a 50 mL volumetric flask. Approximately 48 mL of purified water was added to completely dissolve the mixture, and the solution was adjusted to pH 5.0 with 1 M hydrochloric acid or 1 M NaOH aqueous solution. Purified water was added to the volume. 0.56 g of SIF powder was added and completely dissolved.

[0085] Synthesis of the free form of formula (I)

[0086] Preparation of N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide [ka]

[0087] Prepared with reference to the document of patent application number WO2019154133.

[0088] Step 1): 10-Chloro-5-methoxy-2,3-dihydro-[1,4]dioxano[2,3-f]quinoline (251 mg, 1 mmol) was dissolved in dichloromethane. 1 M boron tribromide in dichloromethane (3 mL, 3 mmol) was added dropwise, and the mixture was stirred until the reaction was complete. The product was concentrated to give 236 mg of a pale yellow solid (5-hydroxy-10-chloro-2,3-dihydro-[1,4]dioxano[2,3-f]quinoline) in 99% yield. MS: 238 [M+H] + .

[0089] Step 2: The product obtained in Step 1 (236 mg, 1 mmol) was dissolved in N,N-dimethylformamide. 4-(3-Chloropropyl)morpholine (163 mg, 1 mmol) and potassium carbonate (414 mg, 3 mmol) were added, and the mixture was heated and stirred until the reaction was complete. Water and ethyl acetate were added to extract the mixture. The organic phase was concentrated and then purified by column chromatography to obtain 291 mg of an off-white solid (10-chloro-5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]-quinoline) in 80% yield. 1 H NMR (400 MHz, DMSO-d 6) δ 8.50 (d, J = 4.8 Hz, 1H), 7.37 (d, J = 4.8 Hz, 1H), 7.10 (s, 1H), 4.47-4.30 (m, 4H), 4.17 (t, J = 6.4 Hz, 2H), 3.59 (t, J = 4.6 Hz, 4H), 2.45 (t, J = 7.1 Hz, 2H), 2.39 (d, J = 4.5 Hz, 4H), 1.97-1.95 (m, 2H). MS: 365[M+H] + .

[0090] Step 3: A reaction flask was charged with chlorobenzene (5 mL), 10-chloro-5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinoline (291 mg, 0.8 mmol), 2-fluoro-4-nitrophenol (125 mg, 0.8 mmol), and triethylamine (0.3 mL), and the mixture was refluxed for 15–20 h. The reaction mixture was cooled and concentrated, and the resulting solid was washed with aqueous potassium carbonate to give 370 mg of a yellow solid product (10-(2-fluoro-4-nitrophenoxy)-5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinoline) in 96% yield. MS: 486 [M+H] + .

[0091] Step 4: The product from Step 3 (370 mg, 0.76 mmol) was added to Pd / C in methanol (10 mL). After purging with hydrogen, the reaction mixture was stirred at room temperature under a hydrogen atmosphere for 10 hours. The reaction mixture was filtered and concentrated to give 340 mg of the product (3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxane[2,3-f]quinolin-10-yl)oxy)aniline) as a white solid. 1 H NMR (400 MHz, DMSO-d 6) δ 8.37 (d, J = 5.3 Hz, 1H), 7.08-6.88 (m, 2H), 6.60-6.49 (m, 1H), 6.48-6.40 (m, 1H), 6.32 (d, J = 5.2 Hz, 1H), 5.44 (s, 2H), 4.37-4.39 (m, 4H), 4.16 (t, J = 6.4 Hz, 2H), 3.59 (t, J = 4.6 Hz, 4H), 2.46 (d, J = 7.0 Hz, 2H), 2.39 (s, 4H), 1.95-1.97 (m, 2H); MS: 456[M+H] + .

[0092] Step 5: A solution of 1-(4-fluorophenylcarbamoyl)cyclopropane-1-carboxylic acid (170 mg, 0.76 mmol) in thionyl chloride (5 mL) was heated to reflux and reacted. After clarification, the refluxing reaction was continued for 1 hour. The resulting reaction mixture was cooled and concentrated to give 180 mg of a yellow solid product (1-(4-fluorophenylcarbamoyl)cyclopropane-1-carbonyl chloride), which was used directly in the next step.

[0093] Step 6): To a solution of 3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)aniline (340 mg, 0.75 mmol) in NMP (2 mL) was added a solution of 1-(4-fluorophenylcarbamoyl)cyclopropane-1-carbonyl chloride (180 mg, 0.75 mmol) in dichloromethane (1 mL) and a solution of triethylamine (0.2 mL), respectively. The mixture was stirred at room temperature for 5 hours and quenched with water. The mixture was washed with saturated sodium carbonate, extracted with dichloromethane, dried, concentrated, and purified by column chromatography to give 260 mg of the product (N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide). 1 H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 10.00 (s, 1H), 8.41 (d, J = 5.2 Hz, 1H), 7.98 - 7.79 (m, 1H), 7.67 - 7.59 (m, 2H), 7.53 - 7.39 (m, 1H), 7.24 (t, J = 9.0 Hz, 1H), 7.18 - 7.11 (m, 2H), 7.06 (s, 1H), 6.43 - 6.34 (m, 1H), 4.37-4.34 (m, 4H), 4.17 (t, J = 6.4 Hz, 2H), 3.59 (t, J = 4.6 Hz, 4H), 2.46 (t, J = 7.1 Hz, 2H), 2.39 (d, J = 4.6 Hz, 4H), 2.08 - 1.79 (m, 2H), 1.47 (d, J = 2.3 Hz, 4H); 13C NMR (101 MHz, DMSO-d6) δ 168.7, 168.4, 160.9, 151.8, 149.6, 146.6, 138.2, 133.8, 129.8, 127.7, 123.4, 122.9, 115.6, 115.4, 107.9, 102.2, 67.1, 66.7, 64.4, 63.97, 55.2, 53.8, 32.3, 26.2, 15.7; MS: 661[M+H] + .

[0094] Example 1. Preparation of crystalline form AI of the mucilage salt of the compound of formula (I)

[0095] [Table 2]

[0096] 300 mg of the compound of formula (I) and mucic acid (acid-base molar ratio 0.5:1) were suspended in acetone and stirred at room temperature for 4 days, and then 15 mg of the compound of formula (I) was added, and the mixture was stirred at room temperature for another 2 days.The mixture was then vacuum dried at room temperature to obtain the product.

[0097] 1 1 H NMR results confirmed that the acid-base molar ratio in the sample was 0.5:1.

[0098] 1HNMR (400 MHz, DMSO-d6) δ 10.34 (s, 1H), 10.02 (s, 1H), 8.42-8.41 (d, J = 4 Hz, 1H), 7.88-7.84(m, 1H), 7.66-7.62 (m, 2H), 7.47-7.44 (m, 1H), 7.26-7.13 (m, 3H), 7.07 (s, 1H), 6.40-6.39 (d, J = 4 Hz, 1H), 4.35 (s, 4H), 4.21 (s, 1H), 4.19-4.16 (t, 2H), 3.77 (s, 1H), 3.61-3.59(m, 4H), 2.43-2.41(m, 6H), 2.01-1.95 (m, 2H), 1.49-1.44 (m, 4H).

[0099] The crystalline form was identified as crystalline form AI by powder X-ray diffraction detection. The XRPD pattern is shown in Figure 1. The characteristic peak positions are shown in Table 2 above, and the TGA and DSC spectra are shown in Figures 2 and 3. The TGA / DSC results showed that the sample had a weight loss of 1.1% when heated to 150°C and had an endothermic peak at 207.3°C (peak temperature).

[0100] Example 2. Preparation of crystalline form A-II of the p-toluenesulfonate salt of compound of formula (I)

[0101] 300 mg of the compound of formula (I) and p-toluenesulfonic acid (acid-base molar ratio 1:1) were suspended in acetone, stirred at room temperature for 1 day, and then vacuum dried at room temperature to obtain a product.

[0102] 1 1 H NMR results confirmed that the acid-base molar ratio in the sample was 1.0:1.

[0103] 1H NMR (400 MHz, MeOD) δ 8.43-8.42 (d, J = 4 Hz, 1H), 7.86-7.82 (m, 1H), 7.70-7.68 (m, 2H), 7.60-7.56 (m, 2H), 7.43-7.40 (m, 1H), 7.25-7.20 (m, 3H), 7.12-7.07 (m, 3H), 6.57-6.56 (m, 1H), 4.44 (s, 4H), 4.37-4.34 (m, 2H), 3.97 (m, 4H), 3.44-3.38(m, 6H), 2.42-2.39 (m, 2H), 2.36 (s, 3H), 1.66 (s, 4H). (active hydrogen on amide not shown)

[0104] The crystalline form was identified as crystalline form A-II by powder X-ray diffraction detection. The XRPD pattern is shown in Figure 4. The characteristic peak positions are shown in Table 3 below, and the TGA and DSC spectra are shown in Figures 5 and 6. The TGA / DSC results showed that the sample had a weight loss of 1.0% when heated to 150°C and had an endothermic peak at 156.2°C (peak temperature).

[0105] [Table 3]

[0106] Example 3. Preparation of crystalline form B-III of the p-toluenesulfonate salt of compound of formula (I)

[0107] [Table 4]

[0108] Approximately 50 mg of the p-toluenesulfonate crystalline form A-II product was suspended in water and stirred at 50° C. for 1 day, filtered, and dried at room temperature to obtain the product.

[0109] 1 1 H NMR results confirmed that the acid-base molar ratio in the sample was 0.9:1.

[0110] 1 H NMR (400 MHz, MeOD) δ 8.43-8.42 (d, J = 4 Hz, 1H), 7.86-7.82 (m, 1H), 7.70-7.68 (m, 2H), 7.60-7.56 (m, 2H), 7.43-7.40 (m, 1H), 7.25-7.20 (m, 3H), 7.12-7.07 (m, 3H), 6.57-6.56 (m, 1H), 4.44 (s, 4H), 4.37-4.34 (m, 2H), 3.97 (m, 4H), 3.44-3.38(m, 6H), 2.42-2.39 (m, 2H), 2.36 (s, 3H), 1.66 (s, 4H). (active hydrogen on amide not shown)

[0111] The crystalline form was identified as crystalline form B-III by powder X-ray diffraction detection. The XRPD pattern is shown in Figure 7. The characteristic peak positions are shown in Table 4 above, and the TGA and DSC spectra are shown in Figures 8 and 9. The TGA / DSC results showed that the sample had a weight loss of 3.4% when heated to 150°C and had two endothermic peaks at 124.4°C and 139.3°C (peak temperatures).

[0112] Example 4. Preparation of crystalline form A-IV of the di-p-toluenesulfonate salt of the compound of formula (I)

[0113] 300 mg of the compound of formula (I) and p-toluenesulfonic acid (acid-base molar ratio 2:1) were suspended in acetone, stirred at room temperature for 1 day, and then vacuum dried at room temperature to obtain a product.

[0114] 1 1 H NMR results confirmed that the acid-base molar ratio in the sample was 2.0:1.

[0115] 1H NMR (400 MHz, MeOD) δ 8.60-8.58 (d, J = 8 Hz, 1H), 7.97-7.93(m, 1H), 7.66-7.64 (m, 4H), 7.60-7.55 (m, 2H), 7.53-7.50 (m, 1H), 7.42-7.38 (m, 1H), 7.19-7.17 (m, 4H), 7.13-7.01 (m, 3H), 6.86-6.84 (m, 1H), 4.56-4.49 (m, 4H), 4.45-4.42 (m, 2H), 4.10 (m, 2H), 3.91(m, 2H), 3.72 (m, 2H), 3.53-3.49 (m, 2H), 3.22 (m, 2H), 2.46-2.43 (m, 2H), 2.34 (s, 6H), 1.61 (s, 4H). (Active hydrogen on amide not shown.)

[0116] The crystalline form was identified as Form A-IV by powder X-ray diffraction detection. The XRPD pattern is shown in Figure 10. The characteristic peak positions are shown in Table 5 below, and the TGA and DSC spectra are shown in Figures 11 and 12. The TGA / DSC results showed that the sample had a weight loss of 1.3% when heated to 150°C and had an endothermic peak at 228.8°C (peak temperature).

[0117] [Table 5]

[0118] Example 5. Preparation of crystalline form AV of the phosphate salt of the compound of formula (I)

[0119] 300 mg of compound of formula (I) and phosphoric acid (acid-base molar ratio 1:1) were mixed in EtOH / HO (19:1, v / v) and 25 mg of seed crystals were added. (The seed crystals were obtained by stirring 50 mg of free compound of formula (I) and phosphoric acid in an equimolar ratio of EtOH / HO (19:1, v / v) at room temperature for 5 days, filtering, and drying at room temperature.) The mixture was suspended and stirred at room temperature for 2 days, and then vacuum dried at room temperature to obtain the product. HPLC / IC results confirmed that the acid-base molar ratio in the sample was 0.8:1.

[0120] 1 H NMR (400 MHz, MeOD) δ 8.40-8.39 (d, J = 4 Hz, 1H), 7.84-7.83 (d, J = 4 Hz, 1H), 7.60-7.56 (m, 2H), 7.41-7.38 (m, 1H), 7.23-7.18 (t, 1H), 7.12-7.07 (m, 2H), 6.52-6.50 (m, 2H), 4.42 (s, 4H), 4.32-4.29 (t, 2H), 3.89-3.87 (m, 4H), 3.09-3.02 (m, 6H), 2.32-2.25 (m, 2H), 1.66 (s, 4H). (Active hydrogen on amide not shown)

[0121] The crystalline form was identified as crystalline form AV by powder X-ray diffraction detection. The XRPD pattern is shown in Figure 13. The characteristic peak positions are shown in Table 6 below, and the TGA and DSC spectra are shown in Figures 14 and 15. The TGA / DSC results showed that the sample had a weight loss of 7.1% when heated to 150°C, and had two endothermic peaks at 90.5°C and 148.2°C (peak temperatures).

[0122] [Table 6]

[0123] Assay Example 1. Determination of the Solubility of Salts and Crystalline Forms of the Present Application in Water

[0124] The dynamic solubilities of the compound of formula (I) in the free state, mucate salt crystalline form AI, p-toluenesulfonate salt crystalline form A-II, di-p-toluenesulfonate salt crystalline form A-IV, and phosphate salt crystalline form AV were evaluated in water and three biological solvents.

[0125] The solubility of each sample in four solvent systems (water, SGF, FaSSIF, and FeSSIF) was measured at 1, 2, 4, and 24 hours using rotary mixing (25 rpm) at 37 °C at a feed concentration of 10 mg / mL (40 mg of material added to 4 mL of solvent). Samples at each time point were centrifuged and filtered (0.45 μm PTFE filter head), and the HPLC concentration and pH of the filtrate were measured. The results of the solubility tests are summarized in Table 7. The results showed that in HO and FaSSIF, the solubilities of p-toluenesulfonate crystalline form A-II, di-p-toluenesulfonate crystalline form A-IV, and phosphate crystalline form AV were significantly improved compared to the free state. In SGF, the solubilities were relatively similar. In FeSSIF, the solubilities of the other four crystalline forms were significantly improved compared to the free state.

[0126] [Table 7]

[0127] LOQ: 0.60 μg / mL; ND: not detected

[0128] The equilibrium solubility of p-toluenesulfonate salt Form A-II and Form B-III in water was also evaluated. Approximately 10 mg / 8 mg of material was weighed into an HPLC vial, and 1.0 mL / 0.8 mL of water was added. The mixture was suspended and stirred at room temperature for 24 hours. The sample was centrifuged and filtered, and the filtrate was tested by HPLC. The solubility results are shown in Table 8. The results indicated that p-toluenesulfonate salt Form A-II was more soluble than Form B-III.

[0129] [Table 8]

[0130] Assay Example 2. Hygroscopicity Test of Salts and Crystal Forms of the Present Application

[0131] The hygroscopicity of the free compound of Formula (I), the mucilage salt crystalline form AI, the p-toluenesulfonate crystalline forms A-II and B-III, the di-p-toluenesulfonate crystalline form A-IV, and the phosphate crystalline form AV was evaluated using a dynamic moisture sorption apparatus (DVS). Starting from 0% relative humidity (0% RH), the mass change rate of the samples was collected at a constant temperature of 25°C while varying the humidity (0% RH - 95% RH - 0% RH). The moisture absorption rates of the six test samples at 25°C / 80% RH are shown in Table 9 below. The results showed that the free compound of Formula (I) is almost non-hygroscopic, the p-toluenesulfonate crystalline form A-II is slightly hygroscopic, and the p-toluenesulfonate crystalline form B-III, the mucilage salt crystalline form AI, the di-p-toluenesulfonate crystalline form A-IV, and the phosphate crystalline form AV are hygroscopic. The crystalline forms of each sample did not change before and after the DVS test.

[0132] [Table 9]

[0133] Assay Example 3. Stability Testing of Salts and Crystalline Forms of the Present Application

[0134] The following six test samples were left under conditions of 25°C / 60% RH and 40°C / 75% RH for one week, and then the physical and chemical stability of the samples was tested by XRPD and HPLC. The test data are shown in Table 10 below. The results showed that the six samples did not undergo any obvious deterioration after being left under conditions of 25°C / 60% RH and 40°C / 75% RH for one week, and that they had good chemical stability and no change in crystal form.

[0135] At the same time, the physical and chemical stability of the solid p-toluenesulfonate salts, Form A-II and Form B-III, was tested by XRPD and HPLC after being stored at 80°C for 24 hours. The results showed that the p-toluenesulfonate salts, Form A-II and Form B-III, did not undergo any change in crystal form or chemical degradation even at such high temperatures.

[0136] [Table 10]

[0137] Long-term stability testing

[0138] The long-term stability of p-toluenesulfonate form A-II of compound of formula (I) was investigated by storing it at 25°C / 60%RH for 6 months. The test data are shown in Table 11 below. The results showed that p-toluenesulfonate form A-II of compound of formula (I) did not undergo any obvious deterioration, had good physical and chemical stability, and its crystalline form did not change even after storing it at 25°C / 60%RH for 6 months.

[0139] [Table 11]

[0140] Assay Example 4. Kinase Activity Testing of Salts and Crystal Forms of the Present Application

[0141] In the present application, the biological kinase activity of the p-toluenesulfonate crystalline form A-II of the free compound prepared in Example 2 was tested and found to be able to effectively inhibit the activity of tyrosine kinases such as PDGFRa, RET, c-KIT, TRK-A, TRK-B, and AXL in addition to VEGFR-2 and c-MET.

[0142] The assay used a Mobility Shift Assay. The starting concentration of the test substance was 2.5 μM, and three-fold serial dilutions were performed to give a total of 10 concentrations in duplicate.

[0143] [Table 12]

[0144] [Table 13]

[0145] Assay Method

[0146] Preparation of 1.1x kinase buffer and stop solution

[0147] 1.1 1x kinase buffer

[0148] 50 mM HEPES, pH 7.5

[0149] 0.0015% Brij-35

[0150] 1.2 Stop liquid

[0151] 100 mM HEPES, pH 7.5

[0152] 0.015% Brij-35

[0153] 0.2% Coating Reagent #3

[0154] 50mM EDTA

[0155] 2. Compound Preparation

[0156] 1) Compound dilution

[0157] The p-toluenesulfonate salt of the free compound prepared in Example 2 was used as the test compound. The starting concentration for the test was 2.5 μM, and this concentration was adjusted to 50 times, i.e., 125 μM. 158 μl of 100% DMSO was added to the second well of the 96-well plate, followed by 2 μl of 10 mM compound solution to prepare a 125 μM compound solution. 60 μl of 100% DMSO was added to the other wells. 30 μl of compound was removed from the second well and added to the third well, and a 3-fold serial dilution was performed, resulting in a total of 10 dilutions. Dilution device: Automatic microwell pipette (Precision PRC384U)

[0158] 2) Transfer 5x compounds to reaction plate

[0159] (1) 10 μl of each well of the 96-well plate containing 50x compound concentration was taken out and transferred to another 96-well plate, to which 90 μl of kinase buffer was added to make the compound concentration 5x.

[0160] (2) 5 μl of each well of the 96-well plate at 5x compound concentration was removed and transferred to a 384-well reaction plate. For example, the compound in well A1 of the 96-well plate was transferred to wells A1 and A2 of the 384-well plate, the compound in well A2 of the 96-well plate was transferred to wells A3 and A4 of the 384-well plate, and so on.

[0161] 3. Kinase Reaction

[0162] 1) Preparation of 2.5x enzyme solution

[0163] Kinase was added to 1x kinase buffer to make a 2.5x enzyme solution.

[0164] 2) Adding enzyme solution to a 384-well plate

[0165] (1) A 384-well reaction plate is pre-filled with 5 μl of 5× compound dissolved in 10% DMSO.

[0166] 10 μl of 2.5x enzyme solution was added to a 384-well reaction plate, and 10 μl of kinase buffer was added to negative control wells.

[0167] (3) Incubate at room temperature for 10 minutes

[0168] 3) Preparation of 2.5x substrate solution

[0169] FAM-labeled peptide and ATP (see Appendix for ATP concentration) were added to 1x kinase buffer to make a 2.5x substrate solution.

[0170] 4) Adding the substrate solution to a 384-well plate

[0171] 10 μl of 2.5x substrate solution was added to a 384-well reaction plate and centrifuged at 1000 rpm for 1 minute.

[0172] 5) Kinase reaction and termination

[0173] (1) The plate was incubated at 28°C for 60 minutes (Biochemical incubator, Model: SPX-100B-Z).

[0174] (2) 30 μl of stopping solution was added to the 384-well reaction plate to stop the reaction, and the plate was centrifuged at 1000 rpm for 1 minute.

[0175] 4. Reading data with Caliper EZ Reader II

[0176] Conversion data was read on a Caliper EZ Reader II.

[0177] 5. Calculation of Inhibition Rate

[0178] 1) Conversion rate data was copied from Caliper EZ Reader II.

[0179] 2) The conversion rates were converted to inhibition rate data, where "Max" refers to the conversion rate of the DMSO control and "Min" refers to the conversion rate of the control with no enzyme activity.

[0180] Percent inhibition = (max-conversion) / (max-min)*100

[0181] 3) IC using XLFit Excel add-in version 5.4.0.8 50 Value fitting

[0182] Fitting formula: Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)^HillSlope)

[0183] [Table 14]

[0184] From this test, test substances for inhibiting the above-mentioned tyrosine kinases were found to be IC 50 was found to be less than 10.0 nM.

[0185] Assay Example 5. Efficacy Test of Different Salts and the Free Compound of the Present Application in Tumor-Bearing Mice

[0186] In this assay, the free compound of formula (I) of the present application and different salts of the compound prepared in Examples 1, 2, and 5 were orally administered to nude mice bearing a tumor model of human lung cancer cell EBC-1, and the effect on tumor growth was examined.

[0187] [Table 15]

[0188] [Table 16]

[0189] Reagents: RPMI 1640 (ThermoFisher, catalog number C11875500BT); fetal bovine serum (Hyclone, catalog number SV30087.03); 0.25% trypsin-EDTA (ThermoFisher, catalog number 25200072); penicillin-streptomycin (Hyclone, catalog number SV30010); DSMO (Life Science, catalog number 0231-500ML); Solutol (Sigma, 70142-34-6-1kg)

[0190] Preparation of test compounds: Appropriate weights of test compounds (including the free form of the compound of formula (I) and different salts of the compounds prepared in Examples 1, 2, and 5) were weighed out and completely suspended / dissolved in an appropriate amount of 0.1% sodium carboxymethylcellulose, stirred and vortexed to obtain a homogenous solution or suspension.

[0191] Methods: All assays were approved by the Animal Welfare Committee. Logarithmic growth phase EBC-1 cells were inoculated subcutaneously into the right dorsal region of immunodeficient nude mice (BALB / c nude, female, 6-8 weeks old, weighing 18±2 g). The cell inoculation volume was 5 × 10 6 The tumor size was 150-200 mm. 3 After reaching maturity, the animals were randomly assigned to groups of 6 animals per group. The doses of each test substance were 15 mg / kg and 7.5 mg / kg (both based on the effective concentration of the compound of formula (I)), and were administered once daily for approximately 3 weeks. The test compounds were prepared fresh every day before use. During the assay period, the diameter of the tumor was measured twice a week, and the body weight of the mice was also measured. The tumor volume (TV) was calculated using the formula TV = 1 / 2 × a × b 2where a and b represent length and width, respectively. Relative tumor volume (RTV) was calculated based on the measurement results. The calculation formula is RTV = Vt / V0. In this formula, V0 is the tumor volume measured at the time of cage grouping and the start of administration (i.e., d0), and Vt is the tumor volume at each measurement. The evaluation index of antitumor activity is the relative tumor growth rate T / C (%), which is calculated as follows: T / C (%) = (TRTV / CRTV) × 100% (TRTV: RTV of the treatment group, CRTV: RTV of the negative control group). T and C represent the average tumor volume at a specific time point in the drug-administered group and the control group, respectively.

[0192] Results: The assay results are shown in Table 17 and Figures 16-19. The different salts of compound of formula (I) and its free compound all showed a certain antitumor effect at doses of 15 mg / kg and 7.5 mg / kg, with the effect being significantly improved at a dose of 15 mg / kg. At the same time, compared with the free compound, all three different salts had superior antitumor effects, with the p-toluenesulfonate salt of compound of formula (I) showing particularly pronounced effects.

[0193] [Table 17]

[0194] The above are alternative embodiments of the present disclosure. It should be noted that a person skilled in the art can improve and modify the embodiments of the present disclosure without departing from the principles of the present disclosure. These improvements and modifications are also intended to fall within the scope of protection of the present disclosure.

Claims

【Request Item 1】 【Chemistry 1】 a pharmaceutically acceptable salt of the compound N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide represented by the formula: Pharmaceutically acceptable salts selected from inorganic and organic salts, alternatively p-toluenesulfonates, mucates, and phosphates.

2. The pharmaceutically acceptable salt according to claim 1, which is a salt of N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide and p-toluenesulfonic acid in a chemical ratio of 1:1 or 1:

2.

3. 2. A method for preparing a pharmaceutically acceptable salt according to claim 1, comprising the step of subjecting N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide to a salt-forming reaction with an inorganic or organic acid, alternatively p-toluenesulfonic acid, mucic acid, or phosphoric acid.

4. The salt-forming reaction is carried out in a solvent selected from water, an alcohol solvent, a halogenated hydrocarbon solvent, a ketone solvent, an ether solvent, a nitrile solvent, an ester solvent, an amide solvent, an aliphatic hydrocarbon solvent, an alicyclic hydrocarbon solvent, an aromatic hydrocarbon solvent, a mixed solvent of an alcohol solvent and water, a mixed solvent of a ketone solvent and water, a mixed solvent of an alcohol solvent and an ether solvent, a mixed solvent of a halogenated hydrocarbon solvent and a nitrile solvent, a mixed solvent of an amide solvent and water, or a mixed solvent of a nitrile solvent and water; the ketone solvent is alternatively acetone, the alcohol solvent is alternatively methanol, ethanol, or isopropanol, the ether solvent is alternatively diethyl ether, methyl tert-butyl ether, tetrahydrofuran, 2-methyltetrahydrofuran, or dioxane, the halogenated hydrocarbon solvent is alternatively dichloromethane or chloroform, the nitrile solvent is alternatively acetonitrile, the ester solvent is alternatively acetone, the alcohol solvent is alternatively methanol, ethanol, or isopropanol, the ether solvent is alternatively diethyl ether, methyl tert-butyl ether, tetrahydrofuran, 2-methyltetrahydrofuran, or dioxane, the halogenated hydrocarbon solvent is alternatively dichloromethane or chloroform, the nitrile solvent is alternatively acetonitrile, the ester solvent is alternatively acetone, the ketone ... the alicyclic hydrocarbon solvent is alternatively cyclohexane; the aromatic hydrocarbon solvent is alternatively toluene, xylene, or isopropylbenzene; the mixed solvent of the alcohol solvent and the ether solvent is alternatively a mixed solvent of diethyl ether and methanol; the mixed solvent of the halogenated hydrocarbon solvent and the nitrile solvent is alternatively a mixed solvent of dichloromethane and acetonitrile; the mixed solvent of the alcohol solvent and water is alternatively a mixed solvent of methanol and water or a mixed solvent of ethanol and water; the mixed solvent of the ketone solvent and water is alternatively a mixed solvent of acetone and water; and the mixed solvent of the amide solvent and water is alternatively a mixed solvent of N,N-dimethylformamide and water.

5. 4. A method for preparing the pharmaceutically acceptable salt of claim 3, comprising the step of forming a salt from N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide and p-toluenesulfonic acid in a chemical ratio of 1:1 or 1:

2.

6. Crystalline Form A-I of mucilage salt of compound N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide of formula (I), wherein the powder X-ray diffraction pattern thereof has characteristic peaks at 2θ angles of 3.95, 5.54, 7.46, 10.87, and 16.58, and the error range of the 2θ angles of each characteristic peak is ±0.2; Alternatively, there are characteristic peaks at 2θ angles of 3.47, 3.95, 5.54, 7.46, 8.46, 10.48, 10.87, 14.90, 15.28, 16.16, and 16.58, and the error range of the 2θ angle of each characteristic peak is ±0.2; Further alternatively, crystalline form AI has characteristic peaks at 2-theta angles of 3.47, 3.95, 5.54, 7.46, 8.46, 10.48, 10.87, 11.72, 14.08, 14.90, 15.28, 16.16, 16.58, 17.90, 20.29, 21.65, 22.35, 23.37, 24.53, 25.45, 27.35, and 32.58, and the error margin of 2-theta angle for each characteristic peak is ±0.

2.

7. Crystalline Form A-II of p-toluenesulfonate salt of N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide of Formula (I), wherein the powder X-ray diffraction pattern thereof exhibits characteristic peaks at 2θ angles of 5.30, 6.54, 7.97, 8.78, 13.01, 15.86, 19.70, 19.94, and 20.62, with the error margin of 2θ angles of each characteristic peak being ±0.2; Alternatively, there are characteristic peaks at 2θ angles of 5.30, 6.54, 7.97, 8.78, 10.52, 11.63, 11.81, 13.01, 15.86, 17.54, 19.34, 19.70, 19.94, 20.62, 22.50, and 25.95, and the error range of the 2θ angle of each characteristic peak is ±0.2; Further alternatively, the 2θ angles are 5.30, 6.54, 7.97, 8.78, 10.52, 11.63, 11.81, 13.01, 13.54, 14.33, 14.52, 15.06, 15.86, 16.46, 17.54, 18.02, 18.25, 18.75, 19.34, 19.70, 19.94, 20.62 , 21.06, 21.46, 22.50, 23.67, 24.02, 24.35, 25.12, 25.95, 26.45, 27.20, 27.87, 29.20, 30.15, 30.87, and 31.83, and the error range of the 2θ angle of each characteristic peak is ±0.

2.

8. Crystalline Form A-IV of di-p-toluenesulfonate salt of N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide of Formula (I), wherein the powder X-ray diffraction pattern thereof exhibits characteristic peaks at 2θ angles of 6.36, 13.60, and 19.20, and the error range of the 2θ angles of each characteristic peak is ±0.2; Alternatively, there are characteristic peaks at 2θ angles of 6.36, 9.87, 11.50, 13.60, 16.00, 19.20, 20.26, 20.86, and 21.36, and the error range of the 2θ angle of each characteristic peak is ±0.2; Further alternatively, crystalline form A-IV has characteristic peaks at 2-theta angles of 6.36, 9.45, 9.87, 11.50, 12.88, 13.60, 14.40, 16.00, 19.20, 20.26, 20.86, 21.36, 22.59, 23.76, and 26.73, and the error margin of 2-theta angle for each characteristic peak is ±0.

2.

9. Crystalline Forms A-V of the phosphate salt of the compound of formula (I), N-(3-fluoro-4-((5-(3-morpholinopropoxy)-2,3-dihydro-[1,4]dioxano[2,3-f]quinolin-10-yl)oxy)phenyl)-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide, have characteristic peaks at 2θ angles of 5.68, 7.56, and 16.93 in their powder X-ray diffraction pattern, with the error range of the 2θ angles of each characteristic peak being ±0.2; Alternatively, there are characteristic peaks at 2θ angles of 4.03, 5.68, 6.84, 7.56, 9.36, 11.30, 11.94, 16.93, 19.76, and 23.01, and the error range of the 2θ angle of each characteristic peak is ±0.2; Further alternatively, crystalline form AV, wherein characteristic peaks are present at 2θ angles of 4.03, 5.68, 6.84, 7.56, 9.36, 11.30, 11.94, 12.56, 13.69, 16.17, 16.93, 19.76, 22.02, 23.01, 23.98, and 25.19, and the error margin of 2θ angle for each characteristic peak is ±0.

2.

10. A pharmaceutical composition comprising a pharmaceutically acceptable salt according to any one of claims 1 to 2 or a crystalline form according to any one of claims 6 to 9, and one or more pharmaceutically acceptable carriers or excipients.

11. 11. The pharmaceutical composition of claim 10, further comprising one or more additional therapeutic agents.

12. Use of a pharmaceutically acceptable salt according to any one of claims 1 to 2, a crystalline form according to any one of claims 6 to 9, or a pharmaceutical composition according to claims 10 to 11 in the manufacture of a medicament for treating a disease associated with the tyrosine kinase VEGFR-2, c-MET, c-KIT, PDGFRa, RET, AXL, or NTRK.

13. 13. The use according to claim 12, wherein the disease is cancer or an autoimmune disease, in particular ocular fundus diseases, xerophthalmia, psoriasis, vitiligo, dermatitis, alopecia areata, rheumatoid arthritis, colitis, multiple sclerosis, systemic lupus erythematosus, Crohn's disease, atherosclerosis, pulmonary fibrosis, liver fibrosis, myelofibrosis, non-small cell lung cancer, small cell lung cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, ovarian cancer, cervical cancer, colorectal cancer, melanoma, endometrial cancer, prostate cancer, bladder cancer, leukemia, gastric cancer, liver cancer, gastrointestinal stromal tumor, thyroid cancer, chronic granulocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, nasopharyngeal carcinoma, esophageal cancer, brain tumor, B-cell and T-cell lymphoma, lymphoma, multiple myeloma, bile duct carcinosarcoma or cholangiocarcinoma.

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