EP4 antagonist compounds, their salts, crystalline polymorphs and uses

Abstract

The present invention relates to an EP4 antagonist represented by formula (I), specifically including its crystalline form, pharmaceutically acceptable salt or crystalline form thereof, methods for preparing them, compositions containing them, and pharmaceutical uses of related compounds. The structure of formula I is shown below: [Case 1] JPEG2024537564000027.jpg40169

Description

Detailed Description of the Invention

[0001] This application claims priority from a prior application filed by the applicant with the State Intellectual Property Office of the People's Republic of China on September 3, 2021, bearing patent number 202111034159.1 and entitled "EP4 antagonist compounds and salts thereof, crystalline polymorphs and uses thereof," the entire text of which is incorporated herein by reference. [Technical field] The present invention belongs to the pharmaceutical field and relates to an EP4 antagonist compound and its salts, crystalline polymorphs, and methods for preparing and using the same. [Background technology] Prostaglandin E2 (PGE2) is an endogenous biologically active lipid. PGE2 activates prostaglandin receptors, triggering a wide range of upstream and downstream dependent biological responses (Legler, DF et al., J. Biochem. Cell Biol. 2010, 42, pp. 198-201). It is involved in the regulation of numerous physiological and pathological processes, including inflammation, pain, renal function, cardiovascular system, pulmonary function, and cancer. PGE2 has been reported to be highly expressed in various cancer tissues, and its correlation with the onset, growth, and progression of cancer and disease states in patients has been confirmed. PGE2 is associated with cell proliferation and activation of cell death (apoptosis) and is generally believed to play an important role in the processes of cancer cell proliferation, disease progression, and cancer metastasis.

[0002] There are four subtypes of PGE2 receptors, EP1, EP2, EP3, and EP4, which are widely distributed in various tissues. Among these subtypes, PGE2 is involved in inflammatory responses (including immunoinflammatory responses), smooth muscle relaxation, pain, lymphocyte differentiation, mesangial cell hypertrophy or proliferation, gastrointestinal mucus secretion, etc. via the EP4 receptor. Therefore, EP4 receptor antagonists are considered to be promising anti-inflammatory and / or analgesic agents for treating diseases associated with the PGE2-EP4 pathway, such as inflammatory diseases and various painful diseases.

[0003] EP4 is a major receptor involved in joint inflammatory pain in rodent models of rheumatoid arthritis and osteoarthritis (see, for example, J. Pharmacol. Exp. Ther., 325, 425 (2008)). Activation leads to the accumulation of the intracellular signal molecule cAMP. Studies have detected EP4 receptor expression in pain receptor peripheral nerve endings, macrophages, and neutrophils, demonstrating the importance of these cell types in endometriosis. Studies have reported that oral EP4 antagonists can reduce proteinuria and inhibit the progression of diabetic nephropathy in type 2 diabetic mice. Another study has shown that activation of EP4 and increased PGE2 production in the bladder mucosa are important causes of overactive bladder (OAB) in prostatitis, and that intravesical injection of an EP4 antagonist can effectively improve OAB after prostatitis. Therefore, selective EP4 antagonists can be used to treat arthritis pain and arthritis, including endometriosis, diabetic nephropathy, and OAB. Existing arthritis treatments are mainly conventional NSAIDs (nonsteroidal anti-inflammatory drugs) or selective COX-2 inhibitors, which may cause cardiovascular and / or gastrointestinal side effects. Selective EP4 antagonists are less likely to cause cardiovascular side effects.

[0004] PGE2 continuously activates EP receptors (produced in large quantities by tumor cells) in the tumor microenvironment (Ochs et al., J Neurochem. 2016, 136, pp. 1142-1154; Zelenay, S. et al., Cell 2015, 162, pp. 1257-1270), promoting the accumulation and enhancing the activity of multiple immune suppressor cells, including tumor-associated macrophages type 2 (TAMS), Treg cells, and myeloid-derived suppressor cells (MDSCs). One of the key immunosuppressive features of the tumor microenvironment is the abundance of MDSCs and TAMs, which is closely correlated with overall survival in patients with gastric cancer, ovarian cancer, breast cancer, bladder cancer, hepatocellular carcinoma (HCC), head and neck cancer, and other types of cancer. Additionally, PGE2 has been reported to induce immune tolerance by inhibiting the accumulation of antigen-presenting dendritic cells (DCs) in tumors and inhibiting the activation of tumor-infiltrating DCs (Wang et al., Trends in Molecular Medicine 2016, 22, pp. 1-3). All of these PGE2-mediated effects jointly aid tumor cell evasion from immune surveillance. PGE2 plays a crucial role in promoting tumor initiation and progression. Increased expression levels of PGE2 and its related receptors, EP2 and EP4, have been found in various malignant tumors, including colon cancer, lung cancer, breast cancer, and head and neck cancer, and are often closely correlated with poor prognosis (Bhooshan, N. et al., Lung Cancer 101, 88-91). Therefore, selective blockade of the EP2 and EP4 signaling pathways can inhibit tumor initiation and progression by altering the tumor microenvironment and regulating tumor immune cells.

[0005] Current preclinical data indicate that EP2 and EP4 specific antagonists can prevent or inhibit tumor growth to varying degrees in animal models of colon, esophageal, lung, and breast cancer. Among the PGE2 receptor drugs that have entered clinical trials, the EP4 antagonist Grapiprant, developed by Pfizer, has been approved by the FDA for the treatment of canine arthritis and, in 2015, entered a Phase II antitumor clinical study for the treatment of multiple solid tumors, including prostate cancer, non-small cell lung cancer, and breast cancer (De Vito, V. et al., J Pharm Biomed Anal 118, 251-258). Eisai's EP4 antagonist E7046 entered a Phase I clinical study in 2015 and a Phase Ib clinical study in 2017 for the treatment of rectal cancer combined with radiation therapy or chemotherapy. ONO-4578, developed by Ono Pharmaceutical, entered a Phase I clinical study for advanced or metastatic solid tumors in 2017, and a Phase I / II clinical study in 2018 as a single agent or in combination with nivolumab to treat advanced solid tumors.

[0006] Chinese patent application CN202110240347.3 discloses the structure of formula I below:

[0007] [ka]

[0008] The compound of formula I can effectively antagonize EP4 receptor activity and has the potential for wide application in the manufacture of therapeutic agents for EP4-related diseases. Therefore, further investigation of the compound of formula I and its crystalline salts and crystalline forms is of great significance for the development of effective therapeutic agents. [Summary of the Invention] To solve the problems present in the prior art, according to one aspect, the present invention provides a crystalline form of the compound of formula I, the structure of which is shown below.

[0009] [ka]

[0010] In some embodiments, the present invention provides free acid crystalline Form A of the compound of Formula I, wherein the free acid crystalline Form A has an X-ray powder diffraction pattern expressed at diffraction angles 2θ±0.2° having diffraction peaks at 7.88, 11.00, 12.13, 16.10, 19.75, 20.65, 21.04, 22.92, 23.53, and 26.69; or The free acid crystalline form A has diffraction peaks at 7.88, 8.08, 11.00, 12.13, 16.10, 19.75, 20.65, and 23.53. Furthermore, the X-ray powder diffraction pattern of the free acid crystalline form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.88, 8.08, 11.00, 12.13, 13.59, 15.50, 16.10, 19.44, 19.75, 20.65, 21.04, 22.92, 23.53, 25.32, 26.44, and further, the X-ray powder diffraction pattern of the free acid crystalline form A, expressed as a diffraction angle of 2θ±0.2°, has the following positions: 7.88, 8.08, 10.31, 10.49, 11.00, 11.46, 12.13, 12.82, 13.59, 15.29, 15.50, 15.72, 16.10, 18.27, 18.70, 19.04, 19.44, 19.75, 20.65, 21.04, 21.62, 21.97 , 22.27, 22.92, 23.53, 24.22, 25.32, 25.81, 26.21, 26.44, 26.69, 27.67, 28.99, 29.35, 30.17, 31.12, 31.65, 32.18, 33.44, 33.97, 35.78, 36.84, 37.18, and 37.99, and further, the free acid crystalline form A has an XRPD pattern essentially as shown in FIG. 1.

[0011] In some embodiments, the free acid crystalline form A is (1) The TGA curve of the free acid crystalline form A has a weight loss of about 0 to 2%, preferably about 0 to 1% (e.g., 0.84%) at 150.0±3°C. (2) The DSC curve of the free acid crystalline form A has an endothermic peak at 134.0±3°C. (3) The DSC curve of the free acid crystalline form A has one, two, or three characteristics, such as an endothermic peak at 136.8±3°C.

[0012] In some embodiments, the free acid crystalline form A is (1) The TGA curve of the free acid crystalline form A shows a weight loss of about 0.84% ​​at 150.0±3°C. (2) The DSC curve of the free acid crystalline form A has an endothermic peak at 134.0±3°C. (3) The DSC curve of the free acid crystalline form A has one, two, or three characteristics, such as an endothermic peak at 136.8±3°C.

[0013] In some embodiments, the TGA pattern of the free acid crystalline form A is shown in FIG. 2, the DSC pattern of the free acid crystalline form A is shown in FIG. 1 The 1 H NMR spectrum is shown in FIG.

[0014] In some embodiments, the present invention provides free acid crystalline Form B of the compound of Formula I, wherein the free acid crystalline Form B exhibits an X-ray powder diffraction pattern expressed at angles 2θ ± 0.2° having diffraction peaks at 7.38, 8.91, 11.07, 17.85, 18.52, 19.38, 23.05, 26.01, and 26.76; and further, the free acid crystalline Form B exhibits an X-ray powder diffraction pattern expressed at angles 2θ ± 0.2° having diffraction peaks at 7.38, 8.91, 11.07, 12.01, 17.85, 18.52, 19.38, 23.05, 26.01, and 26.76; and further, the free acid crystalline Form B exhibits an X-ray powder diffraction pattern expressed at angles 2θ ± 0.2° having diffraction peaks at 5.56, 6.00, 7.38, 8.91, 11.07, 12.01, 17.85, 18.52, 19.38, 23.05, 26.01, and 26.76. , 11.07, 11.58, 12.01, 13.58, 14.16, 14.78, 17.85, 18.52, 19.38, 22.33, 23.05, 24.64, 26.01, and 26.76. Furthermore, the X-ray powder diffraction pattern of the free acid crystalline form B, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.56, 6.00, 7.3 8, 8.27, 8.91, 11.07, 11.58, 12.01, 13.58, 14.16, 14.78, 15.73, 17.85, 18.52, 19.38, 22.33, 23.05, 24.64, 26.01, 26.76, and further, the free acid crystalline form B has an XRPD pattern essentially as shown in FIG.

[0015] In another aspect, the present invention provides a pharmaceutically acceptable salt of the compound of formula I above, which may be selected from alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts), ammonium salts, or salts formed with an organic base providing a physiologically acceptable cation, such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, zinc hydroxide, betaine, monoethanolamine, caffeine, urea, nicotinamide, isonicotine, dimethylglucosamine, ethylglucosamine, glucosamine, meglumine, lysine, arginine, choline, aqueous ammonia, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, diethanolamine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropylene glycol, tromethamine, diethylamine, imidazole, and the like.

[0016] According to a preferred embodiment of the present invention, the pharmaceutically acceptable salts of the compound of formula I above are the tromethamine salt, the diethylamine salt and the lysine salt of the compound of formula I.

[0017] According to an embodiment of the present invention, those skilled in the art will understand that when the compound of formula I forms a salt with a base, the molar ratio of the compound of formula I to the base may be 5:1 to 1:5, such as 3:1, 2:1, 1:1, 1:1.5, 1:2, 1:2.5, or 1:3. It should be understood that the molar ratio of the compound of formula I to the base is preferably 1:1.

[0018] According to another aspect, the present invention provides a crystalline form of a pharmaceutically acceptable salt of a compound of formula I above.

[0019] In some embodiments, the present invention provides tromethamine salt crystalline Form A of the compound of Formula I, wherein the X-ray powder diffraction pattern of Form A, expressed at diffraction angles of 2θ±0.2°, contains diffraction peaks at 6.03, 9.01, 13.50, 15.06, 18.09, and 24.27; and the X-ray powder diffraction pattern of Form A, expressed at diffraction angles of 2θ±0.2°, contains diffraction peaks at 6.03, 9.01, 13.50, 15.06, 16.03, 18.09, 24.27, 27.28, 30.39, and 36.72; and the XRPD pattern of Form A is essentially as shown in FIG. 6 .

[0020] In some embodiments, the tromethamine salt crystalline form A is (1) The TGA curve of tromethamine salt crystalline form A has a weight loss of about 1.0 to 3.5%, preferably about 2.0 to 3.0% (e.g., 2.41%) at 120.0±3°C. (2) The DSC curve of tromethamine salt crystalline form A has an endothermic peak at 124.5±3°C. (3) Tromethamine salt crystalline form A The DSC curve of has one, two or three features, such as one endothermic peak at 139.2±3°C.

[0021] In some embodiments, the tromethamine salt crystalline form A is (1) The TGA curve of tromethamine salt crystalline form A shows a weight loss of about 2.41% at 120.0±3°C. (2) The DSC curve of tromethamine salt crystalline form A has an endothermic peak at 124.5±3°C. (3) Tromethamine salt crystalline form A The DSC curve of has one, two or three features, such as one endothermic peak at 139.2±3°C.

[0022] In some embodiments, the TGA pattern of the tromethamine salt crystalline form A is shown in FIG. 7, the DSC pattern of the tromethamine salt crystalline form A is shown in FIG. 1The 1 H NMR spectrum is shown in FIG.

[0023] In some embodiments, the present invention provides crystalline Form A of the diethylamine salt of the compound of Formula I, wherein the X-ray powder diffraction pattern of Form A, expressed at 2θ±0.2°, has diffraction peaks at 5.29, 9.79, 10.53, 18.30, 19.61, 19.99, 21.10, 25.33, and 26.45; and further, the X-ray powder diffraction pattern of Form A, expressed at 2θ±0.2°, has diffraction peaks at 5.29, 9.79, 10.53, 18.30, 19.61, 19.99, 21.10, 25.33, and 26.45. and 37.44, and further, the diethylamine salt crystalline form A has an XRPD pattern essentially as shown in FIG. 10.

[0024] In some embodiments, the diethylamine salt crystalline form A is (1) The TGA curve of the diethylamine salt crystalline form A has a weight loss of about 0.50 to 3.00%, preferably about 1.00 to 2.50% (e.g., 1.80%) at 100.0±3°C, and a weight loss of about 2.00 to 5.00%, preferably about 3.00 to 4.50% (e.g., 3.99%) at 210.0±3°C. (2) The DSC curve of crystalline form A of diethylamine salt has one or two characteristics, namely, two endothermic peaks at 104.3±10°C and 121.5±10°C, and in particular, two endothermic peaks at 104.3±5°C and 121.5±5°C.

[0025] In some embodiments, the diethylamine salt crystalline form A is (1) The TGA curve of the diethylamine salt crystalline form A shows a weight loss of about 1.80% at 100.0±3°C and a weight loss of about 3.99% at 210.0±3°C. (2) The DSC curve of crystalline form A of diethylamine salt has one or two characteristics, namely, two endothermic peaks at 104.3±10°C and 121.5±10°C, and in particular, two endothermic peaks at 104.3±5°C and 121.5±5°C.

[0026] In some embodiments, the TGA pattern of the diethylamine salt crystalline form A is shown in FIG. 11, the DSC pattern of the diethylamine salt crystalline form A is shown in FIG. 1 The 1 H NMR spectrum is shown in FIG.

[0027] In some embodiments, the present invention provides crystalline Form A of the lysine salt of the compound of Formula I, wherein the X-ray powder diffraction pattern of Form A, expressed at 2θ±0.2°, has diffraction peaks at 5.12, 10.44, 15.56, 18.07, 19.61, and 21.10; and the X-ray powder diffraction pattern of Form A, expressed at 2θ±0.2°, has diffraction peaks at 5.12, 10.44, 15.56, 18.07, 19.61, 21.10, 24.22, and 32.97; and the XRPD pattern of Form A of the lysine salt is essentially as shown in FIG. 14 .

[0028] According to another aspect, the present invention provides methods for preparing the free acid crystalline form A of the compound of formula I above, including the following methods:

[0029] Method 1: The compound of formula I is added to an organic solvent I, which may be one or more combinations selected from ethyl acetate, dichloromethane, methyl tert-butyl ether, isopropanol, and water, and evaporated at room temperature.

[0030] Method 2: The compound of formula I is crystallized in an organic solvent II, which can be one or a combination of one or more selected from the group consisting of methanol, methyl ethyl ketone, isopropyl acetate, tetrahydrofuran, methyl tert-butyl ether, and dimethylacetamide, by adding an anti-solvent, which can be one or a combination of one or more selected from the group consisting of water, isopropyl ether, toluene, meta-xylene, 4-methylisopropylbenzene, n-pentane, n-heptane, cyclohexane, and methylcyclohexane; if no precipitate is formed during stirring, an anti-solvent is added to the system.

[0031] Method 3: The compound of formula I is suspended in an organic solvent III, which may be one or a combination of two or more selected from n-pentane, toluene, meta-xylene, isopropyl ether, n-hexane, cyclohexane, methylcyclohexane, water, methanol, N,N-dimethylformamide, 2-methyltetrahydrofuran, acetone, methyl acetate, dichloromethane, and acetonitrile, at room temperature, and stirred to crystallize.

[0032] Method 4: The compound of formula I is suspended in an organic solvent IV, which may be one or a combination of two or more selected from methylcyclohexane, isopropylbenzene, water, 1,4-dioxane, dimethylacetamide, tetrahydrofuran, n-hexane, 2-methyltetrahydrofuran, n-pentane, methyl ethyl ketone, isopropyl acetate, toluene, isobutanol, chloroform, and meta-xylene, at 40 to 60°C (for example, 50°C), and stirred to crystallize.

[0033] Method 5: The compound of formula I is crystallized by wet-trituration in an organic solvent V, which can be one or more combinations selected from ethanol, dichloromethane, ethyl acetate, tetrahydrofuran.

[0034] In another aspect, the present invention provides a method for preparing crystalline form B of the free acid compound of formula I, comprising dissolving crystalline form A of the free acid compound of formula I in an organic solvent B1 selected from ketones such as methyl ethyl ketone, methyl isopropyl ketone, acetone, diethyl ketone, dipropyl ketone, diisopropyl ketone, dibutyl ketone, and diisobutyl ketone, followed by gas-liquid diffusion in an organic solvent B2 selected from an alkane organic compound, preferably a C1-C7 alkane organic compound, such as n-pentane, n-heptane, and cyclohexane. In some embodiments, the method provides crystalline form A of the free acid compound of formula I, wherein the free acid form B is obtained by dissolving the free acid form A of the compound of formula I in MIBK (methyl isobutyl ketone) followed by gas-liquid diffusion in an n-pentane atmosphere.

[0035] The present invention further provides a method for preparing a pharmaceutically acceptable salt of a compound of Formula I, comprising mixing the compound of Formula I with a salt-forming agent (e.g., a corresponding base) in a suitable solvent. In some embodiments, the solvent is one or a combination of one or more selected from ethanol, heptane, ethyl acetate, MTBE, acetonitrile, water, and acetone.

[0036] In a further aspect, the present invention provides pharmaceutical compositions comprising one or more of the free acid crystalline forms of the compound of formula I (e.g., free acid crystalline form A, free acid crystalline form B), and pharmaceutically acceptable salts of the compound of formula I (including crystalline forms thereof).

[0037] In a further aspect, the present invention provides use of a free acid crystalline form of the compound of formula I (e.g., free acid crystalline form A, free acid crystalline form B), a pharmaceutically acceptable salt of the compound of formula I (including its crystalline form), or the pharmaceutical composition in the manufacture of a medicament for treating or preventing an EP4-related disease.

[0038] According to an embodiment of the present invention, the EP4-related disease includes at least one selected from inflammatory diseases, pain, cancer, metabolic diseases, and urinary system diseases, the inflammatory disease includes at least one selected from arthritis and rheumatoid arthritis, and the pain includes pain caused by osteoarthritis and pain caused by endometriosis.

[0039] According to an embodiment of the present invention, the free acid crystalline form of the compound of Formula I (e.g., free acid crystalline form A, free acid crystalline form B), a pharmaceutically acceptable salt of the compound of Formula I (including its crystalline form), or the pharmaceutical composition can be used in combination with radiation therapy and / or antibody therapy, wherein the antibody therapy is any one or combination of CTLA4 antibody therapy, PDL1 antibody therapy, and PD1 antibody therapy.

[0040] According to embodiments of the invention, the cancer comprises a solid cancer, hi some embodiments, the cancer comprises breast cancer, cervical cancer, colorectal cancer, endometrial cancer, spongioblastoma, head and neck cancer, kidney cancer, liver cancer, lung cancer, medulloblastoma, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, and urinary tract cancer.

[0041] According to an embodiment of the present invention, the metabolic disease comprises diabetes and the urinary disease comprises overactive bladder.

[0042] According to embodiments of the present invention, the use of a free acid crystalline form of the compound of Formula I described in the present invention (e.g., free acid crystalline form A, free acid crystalline form B), a pharmaceutically acceptable salt of the compound of Formula I (including its crystalline form), or the pharmaceutical composition described above can provide patients in need with better and more effective clinical therapeutic agents or treatment regimens.

[0043] The present invention also relates to a method for treating an EP4-related disease, which comprises administering to a patient a therapeutically effective amount of a free acid crystalline form of the compound of formula I (e.g., free acid crystalline form A, free acid crystalline form B), a pharmaceutically acceptable salt of the compound of formula I (including its crystalline form), or a pharmaceutical formulation comprising the pharmaceutical composition.

[0044] Definitions and Interpretations of Terms Unless otherwise specified, the definitions of groups and terms described in the specification and claims of this application, including their illustrative definitions, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, etc., can be arbitrarily combined or combined with each other. The group definitions and compound structures after such combination and combination should fall within the scope described in the specification of this application.

[0045] Unless otherwise specified, numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, 2 or more means 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. When several numerical ranges are defined or understood as "numbers," it should be understood that the two endpoints of the range, each integer within the range, and each decimal point within the range are described. For example, "numbers from 0 to 10" should be understood to describe not only the integers 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, but also the sums of at least each of these integers with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, and 0.9.

[0046] In this specification and claims, when "about" is used in reference to a certain numerical value, it includes the numerical value itself as well as numerical values ​​within a range around the numerical value that is acceptable in the art, such as numerical values ​​within ±15%, ±10%, and ±5%, etc. For example, "about 10" means that it includes numerical values ​​within the range of 10±1.5, i.e., 8.5 to 11.5, 10±1.0, i.e., 9.0 to 11.0, and 10±0.5, i.e., 9.5 to 10.5.

[0047] The term "patient" refers to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse or primate, and most preferably a human.

[0048] The term "therapeutically effective amount" refers to an amount of an active compound or drug that elicits the biological or medical response that a researcher, veterinarian, physician, or other clinician is looking for in a tissue, system, animal, individual, or human, including one or more of the following: (1) disease prevention: e.g., prevention of a disease, disorder, or condition in an individual who is susceptible to the disease, disorder, or condition but has not yet experienced or developed the pathology or symptoms of the disease; (2) disease inhibition: e.g., inhibition of a disease, disorder, or condition (i.e., prevention of further progression of the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition; (3) disease mitigation: e.g., alleviation of a disease, disorder, or condition (i.e., reversal of the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition.

[0049] The term "pharmaceutically acceptable" means that the formulation or active ingredient does not have an undue adverse effect on the health of the general therapeutic target.

[0050] The term "pharmaceutically acceptable excipient or carrier" refers to one or more compatible solid or liquid fillers or gel substances that must be of sufficient purity and sufficiently low toxicity to be suitable for human use. "Compatibility" here means that the components of the composition can be mixed with the compounds of the present invention and with each other without appreciably reducing the efficacy of the compounds. Examples of pharmacologically acceptable excipient or carrier moieties include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil), polyols (e.g., propylene glycol, glycerol, mannitol, sorbitol), emulsifiers, wetting agents (e.g., sodium lauryl sulfate), colorants, flavor enhancers, stabilizers, antioxidants, preservatives, pyrogen-free water, etc. Beneficial effects 1) The present invention provides crystalline forms of the compound of Formula I that have good pharmaceutical properties, of which the free acid crystalline form A has relatively low hygroscopicity and good physical and chemical stability, which is advantageous for drug storage, quality stability, and further formulation.

[0051] 2) The present invention provides medicinal salts of the compound of formula I through optimization measurement screening, and also provides crystalline salt products, such as tromethamine salt crystalline form A, diethylamine salt crystalline form A, and lysine salt crystalline form A, of which tromethamine salt crystalline form A has relatively low hygroscopicity and good physical and chemical stability, and has higher solubility in biological solvents and better pharmaceutical value. [Brief description of the drawing] [Figure 1] XRPD pattern of the free acid crystalline form A of the compound of formula I. [Figure 2] TGA pattern of the free acid crystalline form A of the compound of formula I. [Figure 3] DSC pattern of the free acid crystalline form A of the compound of formula I. [Figure 4] Free acid crystalline form A of the compound of formula I 1 1 H NMR spectrum. [Figure 5] XRPD pattern of the free acid crystalline form B of the compound of formula I. [Figure 6] XRPD pattern of crystalline form A of the tromethamine salt of the compound of formula I. [Figure 7] TGA pattern of crystalline form A of the tromethamine salt of the compound of formula I. [Figure 8] DSC pattern of crystalline form A of the tromethamine salt of the compound of formula I. [Figure 9] Crystalline Form A of Tromethamine Salt of Compound of Formula I 1 1 H NMR spectrum. [Figure 10] XRPD pattern of crystalline form A of the diethylamine salt of the compound of formula I. [Figure 11] TGA pattern of crystalline form A of the diethylamine salt of the compound of formula I. [Figure 12] DSC pattern of crystalline form A of the diethylamine salt of the compound of formula I. [Figure 13] Crystalline Form A of the Diethylamine Salt of Compound of Formula I 1 1 H NMR spectrum. [Figure 14] XRPD pattern of crystalline form A of the lysine salt of the compound of formula I. [Figure 15] Dynamic solubility curve at 37°C. [Figure 16] XRPD overlay of a solubility sample of free acid crystalline form A in HO.

[0033] FIG. 17 is an XRPD overlay of a solubility sample of free acid crystalline form A in SGF.

[0033] FIG. 18 is an XRPD overlay of a solubility sample of free acid Form A in FaSSIF. [Figure 19] XRPD overlay of a solubility sample of free acid Form A in FeSSIF.

[0033] FIG. 20 is an XRPD overlay of a solubility sample of tromethamine salt crystalline form A in SGF. [Figure 21] XRPD overlay of a solubility sample of tromethamine salt crystalline form A in FeSSIF. [Figure 22] DVS pattern of free acid crystalline form A. [Figure 23] XRPD overlay of free acid crystalline form A before and after DVS measurement. [Figure 24] DVS pattern of tromethamine salt crystalline form A. [Figure 25] XRPD overlay of tromethamine salt crystalline form A before and after DVS measurement. [Figure 26] XRPD overlay of a stability evaluation sample of free acid crystalline form A. [Figure 27] XRPD overlay of a stability evaluation sample of tromethamine salt crystalline form A. [Mode for Carrying Out the Invention] The technical solutions of the present invention will be described in more detail below with reference to specific examples. It should be understood that the following examples are merely for illustrative purposes and should not be construed as limiting the scope of the claims of the present invention. Any technology realized based on the above content of the present invention is included within the scope of the claims of the present invention.

[0052] Unless otherwise specified, all raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0053] The instruments and detection methods used in the present invention are as follows: 1. X-ray powder diffraction (XRPD) The XRPD patterns were collected on a PANalytacal X-ray powder diffraction analyzer, and the scan parameters are shown in Table A-1 below.

[0054] [Table 1]

[0055] 2. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) TGA and DSC patterns were collected on a TA Q5000 / 5500 thermogravimetric analyzer and a TA Q2000 / 2500 differential scanning calorimeter, respectively, and the measurement parameters are listed in Table A-2 below.

[0056] [Table 2]

[0057] 3. Dynamic Water Sorption (DVS) Dynamic moisture sorption (DVS) curves were collected using a Surface Measurement Systems (SMS) DVS IntrInsic. Relative humidity at 25°C was corrected for the deliquescence points of LiCl, Mg(NO3)2, and KCl. DVS measurement parameters are listed in Table A-3 below.

[0058] [Table 3]

[0059] 4. Liquid nuclear magnetism ( 1 H NMR) Liquid-state nuclear magnetic resonance spectra were collected on a Bruker 400M nuclear magnetic resonance spectrometer using CD3OD as the solvent.

[0060] 5. High-Performance Liquid Chromatography (HPLC) The purity, dynamic solubility and stability measurements were carried out using an Agilent 1260 high performance liquid chromatograph, and the analytical conditions are shown in Table A-4 below.

[0061] [Table 4]

[0062] The reagents used in the present invention are shown in Table A-5 below.

[0063] [Table 5]

[0064] Example 1 Preparation of Intermediate A

[0065] [ka]

[0066] The starting material, 5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formic acid (5 g, 23.8 mmol) (synthesized in accordance with patent application WO2011151369A1) was added to DCM (dichloromethane) (200 mL) at room temperature, and (S)-methyl 4-(1-aminoethyl)benzoate (5.1 g, 28.6 mmol), HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate) (10.9 g, 28.6 mmol), and DIPEA (N,N-diisopropylethylamine) (4.6 g, 35.7 mmol) were added. The mixture was stirred at room temperature for 16 h, and water (200 mL) was added. DCM (50 The mixture was extracted with 1 mL of hexane (1 mL x 3), the layers were separated, and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by separation on a silica gel column (petroleum ether:ethyl acetate (V / V) = 3:1) to obtain a white solid, methyl (S)-4-(1-(5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formamido)ethyl)benzoate (intermediate A) (5.6 g, yield 63.3%).

[0067] LCMS (ESI) m / z: 372.5 [M+H] + Example 2 Preparation of Compounds of Formula I

[0068] [ka]

[0069] Step 1: 2-(3-(1,1-difluoroethyl)phenyl) pinacorborate (compound IB)

[0070] [ka]

[0071] 1-Bromo-3-(1,1-difluoroethyl)benzene (800 mg, 3.62 mmol) was added to 1,4-dioxane (30 mL) at room temperature, and pinacol diboric acid ester (17.0 g, 156.3 mmol), cuprous iodide (2.5 g, 13.0 mmol), L-proline (2.76 g, 10.86 mmol), potassium acetate (710 mg, 7.24 mmol), and [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride (295 mg, 0.36 mmol) were added. The mixture was heated to 90 °C under nitrogen gas protection and stirred for 16 h. The mixture was cooled to room temperature, diluted with water (200 mL), extracted with dichloromethane (80 mL × 3), and the layers were separated. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column separation (pure petroleum ether) to obtain a colorless liquid crude product, 2-(3-(1,1-difluoroethyl)phenyl)pinacorborate (Compound IB) (900 mg, yield 92.7%).

[0072] Step 2: 3-(1,1-difluoroethyl)phenol (compound IC)

[0073] [ka]

[0074] 2-(3-(1,1-Difluoroethyl)phenyl)pinacorborate (Compound IB) (900 mg, 3.36 mmol) was added to THF (15 mL) and water (15 mL) at room temperature. Sodium perborate monohydrate (1.01 g, 10.07 mmol) was added and stirred at room temperature for 16 h. The mixture was diluted with water (200 mL) and extracted with DCM (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column separation (petroleum ether:ethyl acetate (V / V) = 8:1) to give 3-(1,1-difluoroethyl)phenol (Compound IC) (280 mg, 52.7% yield) as a colorless liquid.

[0075] Step 3: (S)-4-(1-(5-(3-(1,1-difluoroethyl)benzyl)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formamido)ethyl)methyl benzoate (Compound ID)

[0076] [ka]

[0077] The compound (S)-methyl 4-(1-(5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formamido)ethyl)benzoate (Intermediate A) (650 mg, 1.75 mmol) was added to DMF (12 mL) at room temperature, and 3-(1,1-difluoroethyl)phenol (IC) (360 mg, 2.27 mmol) and potassium hydroxide (147 mg, 2.62 mmol) were added, followed by heating to 120°C and stirring for 2 h. The mixture was cooled to room temperature, diluted with water (200 mL), extracted with ethyl acetate (80 mL × 3), and the layers were separated. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a colorless liquid crude product, methyl (S)-4-(1-(5-(3-(1,1-difluoroethyl)benzyl)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formamido)ethyl)benzoate (Compound ID) (1.2 g, crude product).

[0078] LCMS (ESI) m / z: 494.6 [M+H] + .

[0079] Step 4: (S)-4-(1-(5-(3-(1,1-difluoroethyl)benzyl)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formamido)ethyl))benzoic acid (Compound I)

[0080] [ka]

[0081] The starting material (S)-4-(1-(5-(3-(1,1-difluoroethyl)benzyl)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formamido)ethyl)methyl benzoate (Compound ID) (1.0 g, 2.03 mmol) was added to THF (5 mL), water (4 mL), and lithium hydroxide monohydrate (340 mg, 8.11 mmol) were added, and the mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated to produce a white solid, (S)-4-(1-(5-(3-(1,1-difluoroethyl)benzyl)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-formamido)ethyl)benzoic acid (Compound I) (88 mg, 7.9% yield).

[0082] LCMS (ESI) m / z: 480.5 [M+H] + 1 H NMR (400m Hz, DMSO-d6) δ12.8 (s, 1H), 8.10 (d, 1H), 7.71 (d, 2H), 7.53 (t, 1H), 7.47 (d, 1H), 7.27 (d, 1H), 7.11 (t, 1H), 7.11 (d, 2H), 7.07 (dd, 1H), 4.88 (t, 1H), 3.74 (s, 3H), 1.96 (t, 3H), 1.96 (d, 3H). Example 3 Preparation of the free acid crystalline form A of the compound of formula I The compound of formula I prepared according to the method of Example 2 was dissolved in a mixed solvent of dichloromethane and n-hexane (1:10, v / v), and after concentration, the free acid crystalline form A of the compound of formula I was obtained.

[0083] The XRPD of the obtained free acid crystalline form A of the compound of formula I is shown in Figure 1. The TGA / DSC results (Figures 2-3) showed that when the sample was heated to 150°C, it had a weight loss of 0.84% ​​and had one endothermic peak at 136.8±3°C. 1 The H NMR results are shown in Figure 4. Because the free acid crystalline form A had a relatively low TGA weight loss and only a single melting endotherm signal in the DSC, it was inferred to be an anhydrous crystalline form.

[0084] The XRPD analysis data of the obtained free acid crystalline form A of the compound of formula I is shown in Table 3-1 below.

[0085] [Table 6] JPEG0007819298000015.jpg72169

[0086] Example 4 Preparation of the free acid crystalline form B of the compound of formula I The free acid crystalline form A was dissolved in MIBK (methyl isobutyl ketone) until it became transparent, and then subjected to gas-liquid diffusion in an n-pentane atmosphere (its XRPD pattern is shown in FIG. 5).

[0087] The free acid crystalline form B was dried at room temperature and then converted to the free acid crystalline form A.

[0088] The XRPD analysis data of the obtained free acid crystalline form B of the compound of formula I is shown in Table 4-1 below.

[0089] [Table 7]

[0090] Example 5 Preparation and Screening of Salt Forms of Compounds of Formula I Approximately 20 mg of the starting free acid crystalline Form A sample and an equimolar amount of the salt-forming formulation (i.e., the base that forms a salt with the free acid) were weighed into an HPLC vial, and 0.5 mL of solvent was added and mixed to obtain a suspension. The salt-forming formulation was first diluted with the appropriate solvent and then mixed with the starting sample. After approximately 3 days of suspension and stirring at room temperature, the solid was separated by centrifugation and dried overnight in vacuo at room temperature. For colloid-forming systems at room temperature, crystallization was induced by cycling the temperature from 50°C to 5°C. For clear systems at room temperature, crystallization was induced with stirring at 5°C or -20°C. The XRPD characterization results of the resulting solids indicated that a total of three salt forms were obtained in the salt screening assay (shown in Table 5-1 below).

[0091] [Table 8] JPEG0007819298000018.jpg210169

[0092] Example 6 Preparation of Crystalline Form A of the Tromethamine Salt of the Compound of Formula I The free acid crystalline form A and an equimolar amount of tromethamine were slurried with MTBE at room temperature for 3 days, and the solid was separated by centrifugation and dried under vacuum at room temperature to obtain tromethamine salt crystalline form A of the compound of formula I.

[0093] The XRPD pattern of a sample of tromethamine salt crystalline form A is shown in Figure 6. The TGA / DSC results are listed in Figures 7-8. The TGA result showed that the sample had a weight loss of 2.41% when heated to 120°C, and the DSC result showed that the sample had one endothermic peak at 139.2°C (peak value temperature). 1 H NMR was measured by CD3OD and the results are shown in Figure 9. The results showed that in tromethamine salt crystalline form A, the molar ratio of tromethamine to free acid was 1:1, and no residual MTBE solvent was observed.

[0094] The XRPD analytical data of the obtained crystalline form A of the tromethamine salt of the compound of formula I is shown in Table 6-1 below.

[0095] [Table 9]

[0096] Example 7 Preparation of Crystalline Form A of the Diethylamine Salt of the Compound of Formula I A sample of the free acid crystalline form A and an equimolar amount of diethylamine were slurried in MTBE at room temperature for 3 days, and the solid was separated by centrifugation and dried under vacuum at room temperature to obtain diethylamine salt crystalline form A of the compound of formula I.

[0097] The XRPD pattern of a sample of diethylamine salt crystalline form A is shown in Figure 10. The TGA / DSC results are shown in detail in Figures 11-12. The TGA results showed that the sample had a weight loss of 1.80% when heated to 100.0°C and a weight loss of 3.99% when heated from 100.0°C to 210.0°C, and the DSC results showed that the sample had two endothermic peaks at 104.3°C and 121.5°C (peak values). 1 H NMR was measured by CD3OD and the results are shown in Figure 13. The results showed that in diethylamine salt crystalline form A, the molar ratio of diethylamine to free acid was 1:1, and no residual MTBE solvent was observed.

[0098] The XRPD analysis data of the obtained crystalline form A of the diethylamine salt of the compound of formula I is shown in Table 7-1 below.

[0099] [Table 10]

[0100] Example 8 Preparation of Crystalline Form A of the Lysine Salt of Compound of Formula I A sample of the free acid crystalline form A and an equimolar amount of lysine were slurried in EtOAc at room temperature for 3 days, and the solid was separated by centrifugation and dried under vacuum at room temperature to obtain crystalline form A of the lysine salt of the compound of formula I. The sample was characterized by XRPD (see Figure 14).

[0101] The XRPD analysis data of the obtained crystalline form A of the lysine salt of the compound of formula I is shown in Table 8-1 below.

[0102] [Table 11]

[0103] Example 9 Dynamic Solubility Experiments The solids were mixed at a solid input concentration of 10 mg / mL (40 mg of the solid as the free acid was added to 4 mL of solvent) at 37°C under rotational mixing, and the solubility of each sample was measured at different time points (1, 2, 4, and 24 hours) in four systems: water, SGF, FaSSIF, and FeSSIF. After sampling at each time point, the samples were centrifuged (10,000 rpm) and filtered (using a 0.45 μm PTFE filter head). The HPLC concentration and pH of the filtrate were measured, and XRPD was performed on the solid samples after centrifugation. The solubility measurement results are summarized in Table 9-1, the solubility curve is shown in Figure 15, and the XRPD results of the samples after solubility measurement are shown in Figures 16-21. The results showed that the solubility of tromethamine salt Form A in all solvents was superior to that of the free acid Form A. The crystalline form of the free acid Form A remained unchanged after the solvent solubility measurements, and tromethamine salt Form A converted to the free acid Form A in SGF and FeSSIF after the solubility measurements.

[0104] [Table 12]

[0105] Biological Solvent Preparation Instructions: Preparation of simulated gastric fluid (SGF) 200.3 mg of NaCl and 105.0 mg of Trianaton X-100 were weighed and placed in a 50 mL flask, and purified water was added until the mixture became transparent. 136 μL of concentrated hydrochloric acid (12 M) was added, and the pH was adjusted to 1.8 with 1 M hydrochloric acid or 1 M NaOH solution. The solution was then brought to volume with purified water.

[0106] Preparation of intestinal fluid simulating fasting state (FaSSIF) 0.34 g of anhydrous NaH2PO4, 0.44 g of NaOH, and 0.62 g of NaCl were weighed into a 50 mL flask. Purified water was added until the solution became clear, and the pH was adjusted to 6.5 with 1 M hydrochloric acid or 1 M NaOH solution. Purified water was added to the volume, and 0.11 g of SIF powder was weighed and dissolved until the solution became clear.

[0107] Preparation of fed-state simulated intestinal fluid (FeSSIF) 0.82 mL of glacial acetic acid, 0.40 g of NaOH, and 1.19 g of NaCl were placed in a 50 mL flask. Approximately 48 mL of purified water was added until the mixture became clear, and the pH was adjusted to 5.0 with 1 M hydrochloric acid or 1 M NaOH solution. Purified water was added to the volume, and 0.56 g of SIF powder was weighed and dissolved until the mixture became clear.

[0108] Example 10 Hygroscopicity The hygroscopicity of the free acid crystalline form A and the tromethamine salt crystalline form A was evaluated using a dynamic moisture sorption spectrometer (DVS). Starting from 0% RH, the measurements were conducted at a constant temperature of 25°C, and the percentage mass change of the samples was recorded as the humidity changed from 0% RH to 95% RH to 0% RH. The DVS measurement results and the XRPD results before and after the DVS measurement are shown in Figures 22 to 25. The results showed that the water adsorption of the free acid crystalline form A sample at 25°C / 80% RH was 0.07%, indicating almost no hygroscopicity, and the water adsorption of the tromethamine salt crystalline form A sample at 25°C / 80% RH was 0.6%, indicating slight hygroscopicity. The crystalline forms of the free acid crystalline form A and the tromethamine salt crystalline form A samples did not change after the DVS measurement.

[0109] Example 11 Solid state stability The free acid crystalline form A and the tromethamine salt crystalline form A were stored in an open state for 1 / 4 week at 25°C / 60%RH and 40°C / 75%RH, respectively, and the physical and chemical stability of the samples was detected by XRPD and HPLC. The purity data are listed in Table 11-1, and the XRPD results are listed in Figures 26-27. The results showed that the HPLC purity of the free acid crystalline form A and the tromethamine salt crystalline form A did not change significantly after storing them under the corresponding conditions, and the crystalline forms did not change.

[0110] [Table 13]

[0111] Example 12 Different processes for preparing the free acid crystalline form A of the compound of formula I The compound of formula I can be prepared according to the process method of Example 2, and the free acid crystalline form A (identified by XRPD) can also be prepared according to the following method.

[0112] 12.1 20 mg of compound of formula I was added to a 5 mL vial, and 0.4 to 1.0 mL of a solvent was added. After dissolution, the solution was filtered, the vial was sealed with a sealing film, and two pinholes were pierced. The solution was allowed to evaporate slowly at room temperature, and the resulting solid was collected. The solvent was one selected from ethyl acetate, dichloromethane, methyl tert-butyl ether, and isopropanol / water (3:1).

[0113] 12.2 20 mg of the compound of Formula I was placed in a 20 mL vial, and 0.4 to 1.0 mL of solvent was added to each vial. The anti-solvent (approximately 5 mL) was added dropwise to the clear solution with stirring (1000 rpm) until a solid precipitated, or the anti-solvent was added dropwise to the clear solution with stirring (1000 rpm) until a solid precipitated, or after the total volume of the added anti-solvent reached 5 mL, the sample without precipitated solid was suspended and stirred at 5°C, and if a solid still did not precipitate, the sample was suspended and stirred at -20°C, and the final clear sample was evaporated at room temperature), and the resulting solid was collected. When the dissolving solvent is methanol, the anti-solvent is water; when the dissolving solvent is methyl ethyl ketone, the anti-solvent is 4-isopropyltoluene or n-heptane; when the dissolving solvent is isopropyl acetate, the anti-solvent is cyclohexane or meta-xylene; when the dissolving solvent is selected from tetrahydrofuran, the anti-solvent is water or toluene; when the dissolving solvent is selected from methyl tert-butyl ether, the anti-solvent is methylcyclohexane; and when the dissolving solvent is selected from dimethylacetamide, the anti-solvent is selected from water.

[0114] 12.3 20 mg of the compound of Formula I was completely dissolved in 0.4-0.6 mL of solvent. The clear solution was added dropwise to 5 mL of anti-solvent with stirring (1000 rpm), and the precipitated solid was collected. The dissolving solvent was one of isopropanol, ethyl acetate, or 2-methyltetrahydrofuran; the anti-solvent was n-heptane; the dissolving solvent was one of acetone, tert-butyl acetate, or chloroform; the anti-solvent was toluene; and the dissolving solvent was acetonitrile; the anti-solvent was water.

[0115] Approximately 20–90 mg of compound of Formula I was weighed into an HPLC glass vial and 0.5 mL of solvent was added. The resulting suspension was magnetically stirred (1,000 rpm) at room temperature for approximately 4 days, and then centrifuged (10,000 rpm, 2 min) to collect the solid. The solvent was selected from n-pentane, meta-xylene, isopropyl ether, cyclohexane, water, methanol / water (1:5), N,N-dimethylformamide / water (1:4), 2-methyltetrahydrofuran / n-hexane (1:5), acetone / n-heptane (1:4), anisole / n-heptane (1:4), methyl acetate / cyclohexane (1:4), methanol / 4-isopropyltoluene (1:4), dichloromethane / methyl n-hexane (1:5), and acetonitrile / toluene (1:4).

[0116] Approximately 20-50 mg of compound of Formula I was weighed into an HPLC glass vial and 0.5 mL of solvent was added. The resulting suspension was stirred at 50 °C for approximately 3 days with magnetic stirring (1,000 rpm), and then centrifuged at 10,000 rpm for 2 min to collect the solid. The solvent was selected from the group consisting of methylcyclohexane, isopropylbenzene, water, 1,4-dioxane / water (1:9), dimethylacetamide / water (1:9), tetrahydrofuran / n-hexane (1:9), 2-methyltetrahydrofuran / cyclohexane (1:4), cyclopentyl methyl ether / n-pentane (1:4), methyl ethyl ketone / isopropylbenzene (1:7), isopropyl acetate / toluene (1:9), isobutanol / 4-isopropyltoluene (1:4), and chloroform / metaxylene (1:4).

[0117] 12.6 Approximately 20 mg of compound of Formula I was weighed into an HPLC glass vial, and 0.5 mL of solvent was added. The resulting suspension was magnetically stirred (1000 rpm) under temperature cycling (50°C to 5°C, 0.1°C / min, two cycles), and centrifuged (10,000 rpm, 2 min) to collect the solid. The solvent was selected from toluene, n-heptane, 4-isopropyltoluene, ethanol / water (1:4), ethyl acetate / methylcyclohexane (1:9), cyclopentyl methyl ether / n-hexane (1:3), chloroform / n-heptane (1:7), and methyl ethyl ketone / meta-xylene (1:9).

[0118] 12.7 Approximately 20 mg of the compound of Formula I was weighed into a grinding bowl, and 0.2 mL of a solvent was added. The sample was ground for 2-3 minutes, and the solid was collected. The solvent was one selected from ethanol, dichloromethane, ethyl acetate, and tetrahydrofuran.

[0119] Although the embodiments of the present invention have been described above, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention are intended to be included within the scope of the claims of the present invention. [Brief explanation of the drawings]

[0120] [Figure 1] 1 is an XRPD pattern of the free acid crystalline form A of the compound of formula I. [Figure 2] 1 is a TGA pattern of the free acid crystalline form A of the compound of formula I. [Figure 3] 1 is a DSC pattern of the free acid crystalline form A of the compound of formula I. [Figure 4] 1 is a 1H NMR spectrum of the free acid crystalline form A of the compound of formula I. [Figure 5] 1 is an XRPD pattern of the free acid crystalline form B of the compound of formula I. [Figure 6] 1 is an XRPD pattern of crystalline form A of the tromethamine salt of the compound of formula I. [Figure 7] 1 is a TGA pattern of crystalline form A of the tromethamine salt of the compound of formula I. [Figure 8] 1 is a DSC pattern of crystalline form A of the tromethamine salt of the compound of formula I. [Figure 9] 1 is a 1H NMR spectrum of crystalline form A of the tromethamine salt of the compound of formula I. [Figure 10] 1 is an XRPD pattern of crystalline form A of the diethylamine salt of the compound of formula I. [Figure 11] 1 is a TGA pattern of crystalline form A of the diethylamine salt of the compound of formula I. [Figure 12] 1 is a DSC pattern of crystalline form A of the diethylamine salt of the compound of formula I. [Figure 13] 1 is a 1H NMR spectrum of the diethylamine salt of the compound of formula I, crystalline form A. [Figure 14] 1 is an XRPD pattern of crystalline form A of the lysine salt of the compound of formula I. [Figure 15] 1 is a diagram of a dynamic solubility curve at 37°C. [Figure 16] 1 is an XRPD overlay of a solubility sample of free acid Form A in HO. [Figure 17] 1 is an XRPD overlay of a solubility sample of free acid Form A in SGF. [Figure 18]1 is an XRPD overlay of a solubility sample of free acid Form A in FaSSIF. [Figure 19] 1 is an XRPD overlay of a solubility sample of free acid Form A in FeSSIF. [Figure 20] 1 is an XRPD overlay of a solubility sample of tromethamine salt crystalline form A in SGF. [Figure 21] 1 is an XRPD overlay of a solubility sample of tromethamine salt crystalline form A in FeSSIF. [Figure 22] 1 is a DVS pattern of the free acid crystalline form A. [Figure 23] 1 is an XRPD overlay of the free acid crystalline form A before and after DVS measurement. [Figure 24] 1 is a DVS pattern of tromethamine salt crystalline form A. [Figure 25] 1 is an XRPD overlay of tromethamine salt crystalline form A before and after DVS measurement. [Figure 26] 1 is an XRPD overlay of a stability assessment sample of free acid crystalline form A. [Figure 27] 1 is an XRPD overlay of a stability evaluation sample of tromethamine salt crystalline form A.

Claims

1. A crystal of a crystalline form of the compound of formula I, characterized in that the structure of the compound is as shown below: 【Chemistry 1】 The crystals are crystalline form A of the free acid of the compound of formula I, The X-ray powder diffraction pattern of the crystals of the free acid crystalline form A, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.88, 11.00, 12.13, 16.10, 19.75, 20.65, 21.04, 22.92, 23.53, and 26.

69.

2. The crystal described in claim 1, wherein the X-ray powder diffraction pattern of the crystal of the free acid crystalline form A, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.88, 8.08, 11.00, 12.13, 16.10, 19.75, 20.65, and 23.

53.

3. The crystal of claim 1, wherein the X-ray powder diffraction pattern of the crystal of the free acid crystalline form A, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.88, 8.08, 11.00, 12.13, 13.59, 15.50, 16.10, 19.44, 19.75, 20.65, 21.04, 22.92, 23.53, 25.32, 26.44, 26.69, and 27.

67.

4. The X-ray powder diffraction pattern of the crystals of the free acid crystalline form A, expressed as a diffraction angle of 2θ±0.2°, is 7.88, 8.08, 10.31, 10.49, 11.00, 11.46, 12.13, 12.82, 13.59, 15.29, 15.50, 15.72, 16.10, 18.27, 18.70, 19.04, 19.44, 19.75, 20.65, 21.04, 21.

2. The crystal according to claim 1, having diffraction peaks at the following positions: 21.62, 21.97, 22.27, 22.92, 23.53, 24.22, 25.32, 25.81, 26.21, 26.44, 26.69, 27.67, 28.99, 29.35, 30.17, 31.12, 31.65, 32.18, 33.44, 33.97, 35.78, 36.84, 37.18, and 37.

99.

5. The crystal of claim 1, wherein the free acid crystalline form A has an XRPD pattern as shown in FIG. 1: [Figure 1] 。

6. The crystals of the free acid crystalline form A are The TGA curve of the crystals of the free acid crystalline form A has a weight loss of 0-2% at 150.0±3°C, and / or 2. The crystal of claim 1, wherein the DSC curve of the crystal of the free acid crystalline form A has an onset of one endothermic peak at 134.0±3°C.

7. The crystals of the free acid crystalline form A are the TGA curve for the crystals of the free acid crystalline form A has a weight loss of 0.84% ​​at 150.0±3°C, and / or 2. The crystalline form of the compound of formula I according to claim 1, wherein the DSC curve of the free acid crystalline form A has one endothermic peak at 136.8±3°C.

8. A crystal of a crystalline form of the compound of formula I, characterized in that the structure of the compound is as shown below: 【Chemistry 2】 The crystals are crystalline form B of the free acid of the compound of formula I, The X-ray powder diffraction pattern of the crystals of the free acid crystalline form B, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.38, 8.91, 11.07, 17.85, 18.52, 19.38, 23.05, 26.01, and 26.

76.

9. The crystal described in claim 8, wherein the X-ray powder diffraction pattern of the crystal of the free acid crystalline form B, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.38, 8.91, 11.07, 12.01, 17.85, 18.52, 19.38, 23.05, 26.01, and 26.

76.

10. The crystal of claim 8, wherein the X-ray powder diffraction pattern of the crystal of the free acid crystalline form B, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.56, 6.00, 7.38, 8.91, 11.07, 11.58, 12.01, 13.58, 14.16, 14.78, 17.85, 18.52, 19.38, 22.33, 23.05, 24.64, 26.01, and 26.

76.

11. The crystal of claim 8, wherein the X-ray powder diffraction pattern of the crystal of the free acid crystalline form B, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.56, 6.00, 7.38, 8.27, 8.91, 11.07, 11.58, 12.01, 13.58, 14.16, 14.78, 15.73, 17.85, 18.52, 19.38, 22.33, 23.05, 24.64, 26.01, and 26.

76.

12. The crystal of claim 8, wherein the free acid crystalline form B has an XRPD pattern shown in FIG. 5: [Figure 5] 。

13. A pharmaceutically acceptable salt of a compound of formula I, The pharmaceutically acceptable salt of the compound of formula I is the tromethamine salt or the diethylamine salt of the compound of formula I, the structure of which is shown below: A pharmaceutically acceptable salt of a compound of formula I, characterized in that: 【Transformation 3】 。

14. A crystal of a crystalline form of a pharmaceutically acceptable salt of the compound of formula I described in claim 13, wherein the crystal is a crystal of tromethamine salt crystalline form A of the compound of formula I, and the X-ray powder diffraction pattern of the crystal of tromethamine salt crystalline form A, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 6.03, 9.01, 13.50, 15.06, 18.09, and 24.

27.

15. The crystal described in claim 14, wherein the X-ray powder diffraction pattern of the crystal of tromethamine salt crystalline form A, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 6.03, 9.01, 13.50, 15.06, 16.03, 18.09, 24.27, 27.28, 30.39, and 36.

72.

16. The crystal of claim 14, wherein the tromethamine salt crystalline form A has an XRPD pattern as shown in FIG. 6: [Figure 6] 。

17. The crystals of tromethamine salt crystalline form A are The TGA curve of the crystals of tromethamine salt crystalline form A has a weight loss of 1.0 to 3.5% at 120.0±3°C, and / or 15. A crystalline form of the crystalline tromethamine salt of the compound of formula I according to claim 14, wherein the DSC curve of the crystalline form A of the crystalline form has an endothermic peak at the onset of 124.5±3°C.

18. The crystals of tromethamine salt crystalline form A are The TGA curve for the crystals of tromethamine salt crystalline form A has a weight loss of 2.41% at 120.0±3°C, and / or 18. The crystalline form of the pharmaceutically acceptable salt of the compound of formula I according to claim 17, wherein the DSC curve of the crystalline form A of the tromethamine salt has one endothermic peak at 139.2±3°C.

19. The crystal of claim 17, wherein the TGA pattern of the crystal of tromethamine salt crystalline form A is shown in FIG. 7, the DSC pattern of the crystal of tromethamine salt crystalline form A is shown in FIG. 8, and the 1 H NMR spectrum of the crystal of tromethamine salt crystalline form A is shown in FIG. [Figure 7] [Figure 8] [Figure 9] 。

20. A crystal of a crystalline form of a pharmaceutically acceptable salt of the compound of formula I according to claim 13, wherein the crystal is a crystal of diethylamine salt crystalline form A of the compound of formula I, and the X-ray powder diffraction pattern of the crystal of diethylamine salt crystalline form A, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.29, 9.79, 10.53, 18.30, 19.61, 19.99, 21.10, 25.33 and 26.

45.

21. The crystal described in claim 20, wherein the X-ray powder diffraction pattern of the crystal of diethylamine salt crystalline form A, expressed at a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.29, 9.79, 10.53, 11.20, 12.52, 14.85, 15.20, 16.22, 16.86, 18.30, 19.61, 19.99, 21.10, 22.12, 22.47, 24.57, 24.85, 25.33, 26.45, 27.39, 28.01, 29.61, 32.13, 35.00, and 37.

44.

22. The crystal of claim 20, wherein the diethylamine salt crystalline form A has an XRPD pattern as shown in FIG. 10: [Figure 10] 。

23. The crystals of diethylamine salt crystalline form A are The TGA curve of the crystals of diethylamine salt crystalline form A has a weight loss of 0.50 to 3.00% at 100.0±3°C and a weight loss of 2.00 to 5.00% at 210.0±3°C; and / or 21. The crystalline form of the pharmaceutically acceptable salt of the compound of formula I according to claim 20, wherein the DSC curve of the crystalline form A of the diethylamine salt has two endothermic peaks at 104.3±10°C and 121.5±10°C.

24. The crystals of diethylamine salt crystalline form A are The TGA curve of the crystals of diethylamine salt crystalline form A has a weight loss of 1.80% at 100.0±3°C and a weight loss of 3.99% at 210.0±3°C; and / or 24. The crystalline form of the pharmaceutically acceptable salt of the compound of formula I according to claim 23, wherein the DSC curve of the crystalline form A of the diethylamine salt has two endothermic peaks at 104.3±5°C and 121.5±5°C.

25. The crystal of claim 23, wherein the TGA pattern of the crystal of diethylamine salt crystalline form A is shown in FIG. 11, the DSC pattern of the crystal of diethylamine salt crystalline form A is shown in FIG. 12, and the 1 H NMR spectrum of the crystal of diethylamine salt crystalline form A is shown in FIG. [Figure 11] [Figure 12] [Figure 13] 。 26. A crystalline form A of a lysine salt of the compound of formula I, characterized in that the structure of the compound is as shown below, wherein the X-ray powder diffraction pattern of the crystalline form A of the lysine salt, expressed at diffraction angles of 2θ±0.2°, has diffraction peaks at 5.12, 10.44, 15.56, 18.07, 19.61, and 21.10: 【Chemistry 4】 。

27. ​​The crystal described in claim 26, wherein the X-ray powder diffraction pattern of the crystal of lysine salt crystalline form A expressed at a diffraction angle of 2θ±0.2° has diffraction peaks at 5.12, 10.44, 15.56, 18.07, 19.61, 21.10, 24.22, and 32.

97.

28. The crystal of claim 26, wherein the lysine salt crystalline form A has an XRPD pattern as shown in FIG. 14: [Figure 14] 。

29. A method for preparing the crystalline form B of the free acid of the compound of formula I according to claim 8, comprising the steps of: The present invention is characterized in that crystals of the free acid crystalline form A of the compound of formula I are dissolved in MIBK (methyl isobutyl ketone), and then subjected to gas-liquid diffusion in an n-pentane atmosphere to obtain crystals of the free acid crystalline form B of the compound of formula I; The crystals of the free acid crystalline form A have an X-ray powder diffraction pattern expressed at a diffraction angle of 2θ±0.2°, which has diffraction peaks at 7.88, 11.00, 12.13, 16.10, 19.75, 20.65, 21.04, 22.92, 23.53, and 26.

69.

30. 1. A pharmaceutical composition comprising: A pharmaceutical composition comprising a crystalline form of the compound of formula I according to any one of claims 1 to 12, a pharmaceutically acceptable salt of the compound of formula I according to claim 13, or a crystalline form of the compound of formula I according to any one of claims 14 to 28.

31. Use of a crystalline form of the compound of formula I according to any one of claims 1 to 12, a pharmaceutically acceptable salt of the compound of formula I according to claim 13, or a crystalline form of the compound of formula I according to any one of claims 14 to 28 in the manufacture of a medicament for treating or preventing an EP4-related disease, The EP4-related disease is at least one selected from inflammatory diseases, pain, cancer, metabolic diseases, and urinary system diseases, the inflammatory disease is at least one selected from arthritis and rheumatoid arthritis, and the pain includes pain due to osteoarthritis or pain due to endometriosis.

32. The use according to claim 31, which can be used in combination with radiation therapy and / or antibody therapy, wherein the antibody therapy is any one or a combination of CTLA4 antibody therapy, PDL1 antibody therapy, and PD1 antibody therapy.

33. 31. Use of the pharmaceutical composition of claim 30 in the manufacture of a medicament for treating or preventing an EP4-related disease, comprising: The EP4-related disease is at least one selected from inflammatory diseases, pain, cancer, metabolic diseases, and urinary system diseases, the inflammatory disease is at least one selected from arthritis and rheumatoid arthritis, and the pain includes pain due to osteoarthritis or pain due to endometriosis.

34. The use according to claim 33, which can be used in combination with radiation therapy and / or antibody therapy, wherein the antibody therapy is any one or combination of CTLA4 antibody therapy, PDL1 antibody therapy, and PD1 antibody therapy.

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