Pharmaceutically acceptable salts of a benzothiophene-substituted fused ring compound, crystalline forms thereof, and uses thereof

Abstract

The present disclosure relates to a pharmaceutically acceptable salt of a benzothiophene-substituted fused ring compound, a crystalline form thereof and uses. Specifically, the present disclosure provides a pharmaceutically acceptable salt of 2-amino-7-fluoro-4-((5S,5aS,6S,9R)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalen-2-yl)benzo[b]thiophene-3-carbonitrile, a crystalline form thereof and a preparation method thereof, and the corresponding salt has good stability and can be better used for clinical treatment.

Description

Technical Field

[0001] This disclosure belongs to the field of pharmaceutical technology and relates to a pharmaceutically acceptable salt of a benzothiophene-substituted fused-ring compound, its crystalline form, and its uses. Background Technology

[0002] RAS (Rapid Acid Spectroscopy) genes are among the most frequently mutated oncogenes in tumors, with approximately 30% of human malignancies associated with RAS gene mutations. The RAS family includes KRAS, NRAS, and HRAS, with KRAS mutations being the most common, accounting for about 85%. KRAS mutations are frequently found in solid tumors, exhibiting high frequency in the three leading causes of cancer death in humans—lung cancer (17%), colorectal cancer (33%), and pancreatic cancer (61%). In KRAS gene mutations, 97% involve mutations at amino acid residues 12 or 13, with G12D being a significant mutation. Data analysis of populations in Europe and America shows that G12D mutations account for 36%, 12%, and 4% of patients with pancreatic cancer, colorectal cancer, and non-small cell lung cancer, respectively.

[0003] Once activated, KRAS regulates various functions of cell proliferation, survival, migration, and metabolism through numerous downstream signaling pathways, including RAF-MEK-ERK, PI3K-AKT-mTOR, and TIAM1-RAc. Mutations in the KRAS gene result in a persistently activated protein, leading to continued activation of downstream signaling pathways and promoting tumorigenesis.

[0004] PCT / CN2023 / 109598 provides a KRAS G12D inhibitor with the chemical name 2-amino-7-fluoro-4-((5S,5aS,6S,9R)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylbridged naphtho[1,8-ab]heptan-2-yl)benzo[b]thiophene-3-carboxynitrile, having the structure shown in Formula 1.

[0005]

[0006] Salt formation can improve certain undesirable physicochemical or biological properties of drugs. Developing salts with superior physicochemical or pharmaceutical properties compared to 2-amino-7-fluoro-4-((5S,5aS,6S,9R)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentazaza-6,9-methylbridgednaphtho[1,8-ab]heptan-2-yl)benzo[b]thiophene-3-carboxynitrile is of great significance. Given the importance of the crystal form and stability of solid drugs for clinical treatment, in-depth research on the polymorphs of pharmaceutically acceptable salts of compound 2-amino-7-fluoro-4-((5S,5aS,6S,9R)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentazaza-6,9-methylbridgednaphtho[1,8-ab]heptan-2-yl)benzo[b]thiophene-3-carboxynitrile is of great significance for developing drugs suitable for industrial production and with good biological activity. Summary of the Invention

[0007] This disclosure provides a pharmaceutically acceptable salt of the compound 2-amino-7-fluoro-4-((5S,5aS,6S,9R)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentazaza-6,9-methylbridged naphtho[1,8-ab]heptan-2-yl)benzo[b]thiophene-3-carboxynitrile, wherein the pharmaceutically acceptable salt is selected from hydrochloride, sulfate, phosphate, methanesulfonate, acetate, citrate, succinate, p-toluenesulfonate, maleate, fumarate, L-malate, L-tartrate, glutarate, and benzoate.

[0008]

[0009] This disclosure also provides a method for preparing a pharmaceutically acceptable salt of a compound of formula 1, comprising the step of reacting the compound of formula 1 with an acid selected from hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, acetic acid, citric acid, succinic acid, p-toluenesulfonic acid, maleic acid, fumaric acid, L-malic acid, L-tartaric acid, glutaric acid, and benzoic acid.

[0010] The solvents used in the salt formation of this disclosure are selected from, but are not limited to, acetone, ethyl acetate, 2-methyltetrahydrofuran, methanol, and 10% water / methanol.

[0011] In some embodiments, the solvent used for salt formation in this disclosure is solvent A, which is selected from acetone, ethyl acetate, and 2-methyltetrahydrofuran.

[0012] In some embodiments, the solvent used for salt formation in this disclosure is solvent B, which is selected from acetone and 2-methyltetrahydrofuran.

[0013] In some embodiments, the solvent used for salt formation in this disclosure is solvent C, which is selected from ethyl acetate and acetone.

[0014] In some embodiments, the solvent used for salt formation in this disclosure is solvent D, which is selected from ethyl acetate, methanol, and 2-methyltetrahydrofuran.

[0015] Furthermore, in an optional embodiment, the method for preparing the aforementioned pharmaceutically usable salt also includes steps such as crystallization, filtration, washing, or drying.

[0016] In an optional embodiment, the chemical ratio of compound of formula 1 to acid is 3:1 to 1:3, including but not limited to 3:1, 2:1, 1:1, 1:2, and 1:3.

[0017] In another embodiment, the chemical ratio of compound of formula 1 to acid is 2:1 to 1:2.

[0018] In an optional embodiment, the chemical ratio of the compound of formula 1 to hydrochloric acid is 1:1 to 1:2.

[0019] In an optional embodiment, the chemical ratio of the compound of formula 1 to sulfuric acid is 1:1 to 1:2.

[0020] In an optional embodiment, the chemical ratio of the compound of formula 1 to phosphoric acid is 1:1 to 1:2.

[0021] In an optional embodiment, the chemical ratio of the compound of formula 1 to methanesulfonic acid is 1:1 to 1:2.

[0022] In an optional embodiment, the chemical ratio of the compound of formula 1 to citric acid is 1:1.

[0023] In an optional embodiment, the chemical ratio of the compound of formula 1 to succinic acid is 1:1.

[0024] In an optional embodiment, the chemical ratio of the compound of formula 1 to p-toluenesulfonic acid is 1:1 to 1:2.

[0025] In an optional embodiment, the chemical ratio of compound of formula 1 to maleic acid is 1:1.

[0026] In an optional embodiment, the chemical ratio of the compound of formula 1 to fumaric acid is 1:1.

[0027] In an optional embodiment, the chemical ratio of compound of formula 1 to L-malic acid is 1:1 to 1:2.

[0028] In an optional embodiment, the chemical ratio of the compound of formula 1 to L-tartaric acid is 1:1.

[0029] In an optional embodiment, the chemical ratio of the compound of formula 1 to glutaric acid is 1:1 to 1:2.

[0030] In an optional embodiment, the chemical ratio of the compound of formula 1 to benzoic acid is 1:1.

[0031] In some embodiments, the hydrochloride salt of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0032] This disclosure also provides a method for preparing the amorphous hydrochloride salt of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding hydrochloric acid ethanol solution, and stirring.

[0033] In some embodiments, the sulfate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0034] This disclosure also provides a method for preparing the amorphous sulfate of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding sulfuric acid ethanol solution, and stirring.

[0035] This disclosure also provides a sulfate crystal form I of the compound shown in Formula 1, whose X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 4.924, 9.872, 17.934, and 21.926.

[0036] In some embodiments, the X-ray powder diffraction pattern of the sulfate crystal form I of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 1 As shown.

[0037] This disclosure also provides a method for preparing sulfate crystal form I of the compound shown in Formula 1, comprising the steps of adding the amorphous compound of Formula 1 to 10% water / methanol, then adding sulfuric acid ethanol solution, and stirring.

[0038] In some embodiments, the phosphate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0039] This disclosure also provides a method for preparing the amorphous phosphate of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding a phosphoric acid ethanol solution, and stirring.

[0040] This disclosure also provides a phosphate crystal form I of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 5.572 and 9.488.

[0041] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystal form I of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 2 As shown.

[0042] This disclosure also provides a method for preparing phosphate crystal form I of the compound shown in Formula 1, comprising the steps of adding the amorphous compound of Formula 1 to 10% water / methanol, then adding a phosphoric acid ethanol solution, and stirring.

[0043] In some embodiments, the methanesulfonate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0044] This disclosure also provides a method for preparing the amorphous methanesulfonate of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding a methanesulfonate ethanol solution, and stirring.

[0045] In some embodiments, the acetate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0046] This disclosure also provides a method for preparing the amorphous acetate of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent B, adding an acetic acid-ethanol solution, and stirring.

[0047] In some embodiments, the citrate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0048] This disclosure also provides a method for preparing the amorphous citrate of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding a citric acid ethanol solution, and stirring.

[0049] In some embodiments, the succinate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0050] This disclosure also provides a method for preparing the amorphous succinate of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding succinic acid, and stirring.

[0051] In some embodiments, the p-toluenesulfonate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0052] This disclosure also provides a method for preparing the amorphous p-toluenesulfonate of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding a p-toluenesulfonic acid ethanol solution, and stirring.

[0053] In some embodiments, the maleate salt of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0054] This disclosure also provides a method for preparing the amorphous maleate salt of the compound shown in Formula 1, comprising the steps of dissolving the compound of Formula 1 in solvent A, adding a maleic acid ethanol solution, and stirring.

[0055] The fumarate crystal form I of the compound of Formula 1 provided in this disclosure has characteristic peaks at 9.184, 15.751, and 19.708 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0056] In some embodiments, the X-ray powder diffraction pattern of the fumarate crystal form I of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 3 As shown.

[0057] This disclosure also provides a method for preparing fumarate crystal form I of the compound shown in Formula 1, the method comprising the steps of dissolving the compound of Formula 1 in solvent A, adding solid fumaric acid, and stirring.

[0058] In some embodiments, the L-malate of the compound shown in Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0059] This disclosure also provides a method for preparing the amorphous form of L-malate of the compound shown in Formula 1, the method comprising the steps of dissolving the compound of Formula 1 in solvent A, adding an L-malic acid ethanol solution, and stirring.

[0060] This disclosure also provides an L-malate crystal form I of the compound shown in Formula 1, whose X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 9.680, 14.709, 15.093, 15.515, and 19.891.

[0061] In some embodiments, the X-ray powder diffraction pattern of the L-malate crystal form I of the compound shown in Formula 1, expressed in terms of diffraction angle 2θ, is as follows: Figure 4 As shown.

[0062] This disclosure also provides a method for preparing L-malate crystal form I of the compound shown in Formula 1, the method comprising the steps of dissolving the compound of Formula 1 in solvent C, adding an L-malic acid ethanol solution, and stirring.

[0063] The L-tartrate crystal form I of the compound of Formula 1 provided in this disclosure has characteristic peaks at 4.959, 9.606, 15.022, and 19.631 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0064] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate crystal form I of the compound shown in Formula 1, expressed in terms of diffraction angle 2θ, is as follows: Figure 5 As shown.

[0065] This disclosure also provides a method for preparing L-tartrate crystal form I of the compound shown in Formula 1, the method comprising the steps of dissolving the compound of Formula 1 in solvent A, adding an L-tartrate ethanol solution, and stirring.

[0066] This disclosure also provides a method for preparing tartrate crystal form I of the compound shown in Formula 1, the method comprising the steps of adding the compound of Formula 1 to 10% water / methanol, then adding L- tartaric acid ethanol solution, and stirring.

[0067] This disclosure also provides an L-tartrate crystal form II of the compound shown in Formula 1, whose X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 5.073, 8.094, 10.179, 13.173, 16.244, 19.316, and 20.966.

[0068] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate crystal form II of the compound shown in Formula 1, expressed in terms of diffraction angle 2θ, is as follows: Figure 6 As shown.

[0069] This disclosure also provides a method for preparing L-tartrate crystal form II of the compound shown in Formula 1, the method comprising the steps of adding the compound of Formula 1 to acetone, then adding an L-tartrate ethanol solution, and stirring.

[0070] The glutarate crystal form I of the compound of Formula 1 provided in this disclosure has characteristic peaks at 10.374, 15.175, and 19.746 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0071] In some embodiments, the X-ray powder diffraction pattern of the glutarate crystal form I of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 7 As shown.

[0072] This disclosure also provides a method for preparing glutarate crystal form I of the compound shown in Formula 1, the method comprising dissolving the compound of Formula 1 in solvent A, then adding glutaric acid and stirring.

[0073] This disclosure also provides a glutarate crystal form II of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 8.490, 13.020, 15.362, 18.433, and 21.619.

[0074] In some embodiments, the X-ray powder diffraction pattern of the glutarate crystal form II of the compound shown in Formula 1, expressed in terms of diffraction angle 2θ, is as follows: Figure 8 As shown.

[0075] This disclosure also provides a method for preparing glutarate crystal form II of the compound shown in Formula 1, the method comprising the steps of adding the compound of Formula 1 to ethyl acetate, then adding glutaric acid, and stirring.

[0076] The benzoate crystal form I of the compound of Formula 1 provided in this disclosure has characteristic peaks at 7.340, 7.878, 12.295, and 19.439 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0077] In some embodiments, the X-ray powder diffraction pattern of the benzoate crystal form I of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 9 As shown.

[0078] This disclosure also provides a method for preparing benzoate crystal form I of the compound shown in Formula 1, the method comprising the steps of dissolving the compound of Formula 1 in solvent A, then adding a benzoic acid ethanol solution, and stirring.

[0079] This disclosure also provides a complex formed by the compound of Formula 1 and a ligand selected from arginine and o-benzoylsulfonylimide.

[0080] In some embodiments, the complex formed by the compound of Formula 1 and o-benzoylsulfonylimide is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0081] This disclosure provides a method for preparing an amorphous compound of o-benzoylsulfonylimide, which is a compound of Formula 1. The method includes dissolving the compound of Formula 1 in solvent D, adding o-benzoylsulfonylimide, and stirring.

[0082] This disclosure also provides a crystalline form I of the o-benzoylsulfonylimide complex of the compound shown in Formula 1, whose X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 6.985, 8.161, 11.381, 16.472, 19.186, 23.303, and 29.367.

[0083] In some embodiments, the crystalline form I of the o-benzoylsulfonylimide complex of the compound shown in Formula 1, as expressed in diffraction angle 2θ, has characteristic peaks at 6.985, 8.161, 11.381, 12.871, 14.733, 15.137, 16.472, 18.232, 19.186, 23.303, 25.891, and 29.367.

[0084] In some embodiments, the X-ray powder diffraction pattern of the benzoylsulfonylimide complex I of Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 6.985, 8.161, 8.953, 9.529, 11.381, 12.871, 14.733, 15.137, 16.472, 18.232, 19.186, 21.129, 21.974, 23.303, 23.971, 25.891, 26.938, 27.466, 29.367, and 30.500.

[0085] In some embodiments, the X-ray powder diffraction pattern of the o-benzoylsulfonylimide complex I of Formula 1, expressed in terms of diffraction angle 2θ, is as follows: Figure 10 As shown.

[0086] This disclosure also provides a method for preparing the o-benzoylsulfonylimide complex of Formula 1, crystal form I, the method comprising adding the compound of Formula 1 to acetone or 10% water / methanol, then adding o-benzoylsulfonylimide, and stirring.

[0087] The arginine complex crystal form a of the compound of Formula 1 provided in this disclosure has characteristic peaks at 15.849, 18.709, 19.847, 20.783, 23.275, 26.583, and 32.305 in its X-ray powder diffraction pattern expressed as a diffraction angle of 2θ.

[0088] In some embodiments, the X-ray powder diffraction pattern of the arginine complex a of the compound shown in Formula 1, expressed as a diffraction angle 2θ, is as follows: Figure 11 As shown.

[0089] This disclosure also provides a method for preparing crystal form a of the arginine complex of Formula 1, the method comprising dissolving the compound of Formula 1 in acetone, adding arginine, and stirring.

[0090] This disclosure also provides a pharmaceutical composition comprising, optionally, the aforementioned hydrochloride, sulfate, phosphate, methanesulfonate, acetate, citrate, succinate, p-toluenesulfonate, maleate, fumarate, L-malate, L-tartrate, glutarate, benzoate or their respective crystal forms, or o-benzoylsulfonylimide complex crystal form I or arginine complex crystal form a, and pharmaceutical excipients optionally selected from pharmaceutically acceptable excipients.

[0091] This disclosure also provides a pharmaceutical composition prepared from the aforementioned hydrochloride, sulfate, phosphate, methanesulfonate, acetate, citrate, succinate, p-toluenesulfonate, maleate, fumarate, L-malate, L-tartrate, glutarate, benzoate or their respective crystal forms, or o-benzoylsulfonylimide complex crystal form I or arginine complex crystal form a, and optionally a pharmaceutically acceptable excipient.

[0092] This disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing optionally the aforementioned hydrochloride, sulfate, phosphate, methanesulfonate, acetate, citrate, succinate, p-toluenesulfonate, maleate, fumarate, L-malate, L-tartrate, glutarate, benzoate or their corresponding crystal forms, or o-benzoylsulfonylimide complex crystal form I or arginine complex crystal form a, with a pharmaceutically acceptable excipient.

[0093] This disclosure also provides the use of the aforementioned hydrochloride, sulfate, phosphate, methanesulfonate, acetate, citrate, succinate, p-toluenesulfonate, maleate, fumarate, L-malate, L-tartrate, glutarate, benzoate or their corresponding crystal forms, or o-benzoylsulfonylimide complex crystal form I or arginine complex crystal form a, or the aforementioned compositions in the preparation for the prevention and / or treatment of cancer.

[0094] The uses described in this disclosure, wherein the cancer is selected from breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, gastric cancer, colorectal cancer, lung cancer, kidney cancer, liver cancer, cervical cancer, endometrial cancer, epithelial cancer, esophageal cancer, neuroblastoma, glioma, bone cancer, nasopharyngeal carcinoma, oral cancer, thyroid cancer, skin cancer, and squamous cell carcinoma; preferably, the cancer is selected from breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, gastric cancer, colorectal cancer, and lung cancer.

[0095] The "2θ or 2θ angle" mentioned in this disclosure refers to the diffraction angle, where θ is the Bragg angle, and the unit is ° or degree; the error range of 2θ for each characteristic peak is ±0.20 (including the case where the number has more than one decimal place after rounding), specifically -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

[0096] The numerical values ​​in this disclosure, such as those relating to the content of certain substances, are calculated data and inevitably contain a certain degree of error. Generally, ±10% is within the reasonable error range. The error may vary to some extent depending on the context in which it is used, but this variation shall not exceed ±10%, and may be ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%, preferably ±5%.

[0097] The starting material used in the crystal form preparation method disclosed herein can be any form of compound, including but not limited to: amorphous, arbitrary crystal form, hydrate, solvate, etc.

[0098] The drying temperature described in this disclosure is generally 25℃-100℃, preferably 40℃-70℃, and can be dried under normal pressure or reduced pressure.

[0099] The crystallization methods described in this disclosure include room temperature crystallization, cooling crystallization, solvent evaporation crystallization, and seed crystallization induction. The cooling temperature is selected from below 65°C, preferably from -10°C to 60°C. Stirring can also be performed during the crystallization process.

[0100] The “differential scanning calorimetry or DSC” described in this disclosure refers to measuring the temperature difference and heat flow difference between the sample and the reference material during the sample heating or isothermal process, in order to characterize all physical and chemical changes related to thermal effects and obtain phase transition information of the sample.

[0101] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "Guiding Principles on Hygroscopicity of Drugs" in Part IV of the 2015 edition of the Chinese Pharmacopoeia,

[0102] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0103] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15%;

[0104] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15% but not less than 2%;

[0105] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0106] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0107] The “excipients” described in this disclosure include, but are not limited to, any adjuvants, carriers, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, or emulsifiers that have been approved by the U.S. Food and Drug Administration for use in humans or livestock. Attached Figure Description

[0108] Figure 1 The image shows the XRPD spectrum of compound 1 sulfate crystal form I.

[0109] Figure 2 The image shows the XRPD spectrum of phosphate form I of compound 1.

[0110] Figure 3 The image shows the XRPD spectrum of compound 1 fumarate crystal form I.

[0111] Figure 4 The image shows the XRPD spectrum of compound 1L-malate crystal form I.

[0112] Figure 5 The image shows the XRPD spectrum of compound 1L-tartrate crystal form I.

[0113] Figure 6 The XRPD spectrum of compound 1L-tartrate crystal form II is shown.

[0114] Figure 7 The image shows the XRPD spectrum of compound 1 glutarate crystal form I.

[0115] Figure 8 The image shows the XRPD spectrum of compound 1 glutarate crystal form II.

[0116] Figure 9 The image shows the XRPD spectrum of benzoate form I of compound 1.

[0117] Figure 10 The image shows the XRPD spectrum of compound 1, o-benzoylsulfonylimide complex, crystal form I.

[0118] Figure 11 The image shows the XRPD spectrum of arginine complex a, a crystal form of compound 1. Detailed Implementation

[0119] The present disclosure will be explained in more detail below with reference to embodiments or experimental examples. The embodiments or experimental examples in the present disclosure are only used to illustrate the technical solutions in the present disclosure and are not intended to limit the substance and scope of the present disclosure.

[0120] Test conditions of the instruments used in the experiment:

[0121] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ ohms. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE NEO 500M NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0122] MS measurements were performed using an Agilent 1200 / 1290DAD-6110 / 6120 Quadrupole MS LC-MS system (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD system (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), or a THERMO Ultimate3000-Q Exactive system (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0123] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent HPLC 1200DAD, an Agilent HPLC 1200VWD, and a Waters HPLC e2695-2489 HPLC system.

[0124] High performance liquid chromatography (HPLC) was performed using Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson-281 preparative chromatographs.

[0125] Silica gel column chromatography generally uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0126] XRPD (X-ray Powder Diffraction) was used for analysis: measurements were performed using a BRUKER D8 X-ray diffractometer. Specific data collected included: Cu anode (40 kV, 40 mA), Cu-Kα1 rays. Kα2 rays Kβ rays Scanning mode: θ / 2θ, scanning range (2θ range): 3°~45°.

[0127] DSC stands for Differential Scanning Calorimetry: Measurements were performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10℃ / min. The specific temperature range was referenced from the corresponding spectra (mostly 25-250℃ or 350℃), and the nitrogen purging rate was 50mL / min.

[0128] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA2 thermogravimetric analyzer, with a heating rate of 10℃ / min, and the specific temperature range was referenced from the corresponding spectrum (mostly 30-350℃). The nitrogen purging rate was 50mL / min.

[0129] DVS stands for Dynamic Moisture Adsorption: The detection method is SMSDVS Advantage. At 25℃, the humidity changes from 50% to 95% to 0% to 95% to 50%, with a step of 10% (the last step is 5%). (The specific humidity range is subject to the corresponding spectrum. The methods listed here are the most commonly used methods.) The judgment standard is that dm / dt is not greater than 0.002%.

[0130] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, and Darui Chemicals.

[0131] Unless otherwise specified in the examples, all reactions can be carried out under an argon or nitrogen atmosphere.

[0132] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0133] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system used for column chromatography to purify the compounds, and the developing solvent system for TLC included: A: n-hexane / ethyl acetate system, B: dichloromethane / methanol system. The volume ratio of the solvent was adjusted according to the polarity of the compounds, and small amounts of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0134] Example 1

[0135] 2-Amino-7-fluoro-4-((5S,5aS,6S,9R)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylbridgednaphtho[1,8-ab]heptan-2-yl)benzo

[0136] [b]Thiophene-3-carboxynitrile 1

[0137]

[0138] first step

[0139] 4-(fluoromethyl)piperidine-1-carboxylic acid tert-butyl ester 1b

[0140] 2-((fluoromethyl)sulfonyl)pyridine (4.2 g, 23.97 mmol) was dissolved in tetrahydrofuran (50 mL). A 1 M solution of bis(trimethylsilyl)amino potassium in tetrahydrofuran (30 mL) was added at -78 °C. After reacting for 30 minutes, N-tert-butoxycarbonyl-4-piperidinone 1a (5 g, 25.09 mmol, Shanghai Shaoyuan) was added. The reaction was maintained at 3 hours, then allowed to return to room temperature for 1 hour. Saturated ammonium chloride was added to quench the reaction mixture, followed by the addition of 3N hydrochloric acid (100 mL). After stirring for 1 hour, the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system B to obtain solid compound 1b (2 g, yield: 37%).

[0141] Step 2

[0142] 4-(Fluoromethyl)piperidine hydrochloride 1c

[0143] Compound 1b (1 g, 4.64 mmol) was dissolved in 20 mL of 4 M hydrogen chloride solution of 1,4-dioxane and stirred for 1 hour. The reaction solution was concentrated under reduced pressure to obtain crude solid compound 1c (700 mg). The product was used directly in the next step of the reaction without purification.

[0144] MS m / z(ESI): 116.1 [M+1].

[0145] Step 3

[0146] 2,5,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine-4-phenol 1e

[0147] The crude compound 1d (2 g, 8 mmol) was dissolved in phosphorus oxychloride (25 mL), and N,N-diisopropylethylamine (5.16 g, 40 mmol) was added. The mixture was stirred at 110 °C for 14 hours. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane, and the pH was adjusted to 2-3 by adding 20% ​​potassium carbonate solution. After stirring for 2 hours, the mixture was filtered. The filter cake was washed with water and dried to obtain the crude solid compound 1e (1.5 g). The product was used directly in the next step without purification.

[0148] MS m / z(ESI):267.8[M+1].

[0149] Step 4

[0150] (1S,2S,5R)-2-((S)-1-((2,7-dichloro-8-fluoro-4-hydroxypyridino[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo

[0151] [3.2.1] 1g of tert-butyl octane-8-carboxylate

[0152] (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester 1f (370 mg, 1.44 mmol, prepared by the method disclosed in Intermediate 29 on page 164 of patent application "WO2022173678A1") was dissolved in tetrahydrofuran (10 mL). Sodium hydride (201 mg, 5.2 mmol, 60% purity) was added under ice bath. After reacting for 30 minutes, compound 1e (353 mg, 1.31 mmol) was added. The reaction was stirred for 2 hours. The reaction solution was quenched with water and concentrated under reduced pressure to obtain 1 g (600 mg) of crude solid compound. The product was used directly in the next step without purification.

[0153] MS m / z(ESI):488.2[M+1].

[0154] Step 5

[0155] (5S,5aS,6S,9R)-2,12-dichloro-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentazaze

[0156] -6,9-methyl-naphtho[1,8-ab]heptan-14-carboxylic acid tert-butyl ester 1h

[0157] 1 g (78 mg, 159.7 μmol) of the compound was dissolved in dichloromethane (2 mL). N,N-diisopropylethylamine (61.9 mg, 478.9 μmol) and phosphorus oxychloride (122.4 mg, 798.2 μmol) were added under ice bath conditions. The mixture was stirred for 2 hours. The reaction solution was quenched with saturated sodium bicarbonate solution. The reaction was then combined with dichloromethane (10 mL × 2). The mixture was dried with anhydrous sodium sulfate. After filtration to remove the desiccant, the product was concentrated under reduced pressure to obtain crude solid compound 1 h (75 mg). The product was used directly in the next reaction without purification.

[0158] MS m / z(ESI): 470.2 [M+1].

[0159] Step 6

[0160] (5S,5aS,6S,9R)-12-((1-(((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)methoxy)-2-chloro-1-fluoro-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylbridgednaphtho[1,8-ab]heptan-14-carboxylic acid tert-butyl

[0161] 1j base ester

[0162] Dissolve 1.4 g (6.4 mmol) of (1-(((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)methanol 1i in tetrahydrofuran (15 mL), add 2 M sodium bis(trimethylsilyl)amino in tetrahydrofuran under ice bath, and stir for 30 minutes while maintaining the temperature. Then add 20 mL of tetrahydrofuran solution of crude compound 1h (2.3 g, 4.9 mmol) under ice bath, and stir for 1 hour while maintaining the temperature. Quench the reaction solution with saturated ammonium chloride solution, extract with ethyl acetate (30 mL × 2), combine the organic phases, dry with anhydrous sodium sulfate, filter to remove the desiccant, concentrate under reduced pressure, and purify the residue by silica gel column chromatography with eluent system B to give solid compound 1j (2 g, yield: 62.8%).

[0163] MS m / z(ESI): 650.2 [M+1].

[0164] Step 7

[0165] (5S,5aS,6S,9R)-2-chloro-1-fluoro-12-((1-(hydroxymethyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa

[0166] -3,10a,11,13,14-pentazaza-6,9-methyl-naphtho[1,8-ab]heptan-4-carboxylic acid tert-butyl ester 1k

[0167] Compound 1j (100 mg, 153.8 μmol) was dissolved in tetrahydrofuran (4 mL), and a 1 M tetrabutylammonium fluoride tetrahydrofuran solution (187 μL) was added. The mixture was stirred for 2 hours, and the reaction solution was quenched with saturated ammonium chloride aqueous solution. The mixture was extracted with ethyl acetate (15 mL × 3), and the organic phases were combined. The mixture was washed successively with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain crude solid compound 1k (82 mg). The product was used directly in the next reaction without purification. MS m / z (ESI): 536.2 [M+1].

[0168] Step 8

[0169] (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-((1-((methanesulfonyl)oxy)methyl)cyclopropyl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylbridged naphtho[1,8-ab]heptan-14-carboxylic acid tert-butyl ester

[0170] 1l

[0171] The crude compound 1k (83 mg, 154.9 μmol) and N,N-diisopropylethylamine (60 mg, 464.2 μmol) were dissolved in dichloromethane (3 mL). Methanesulfonyl chloride (25 mg, 218.2 μmol) was added under ice bath conditions. The reaction was allowed to return to room temperature for 30 minutes. The reaction solution was quenched with saturated ammonium chloride aqueous solution. The mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined and washed successively with water and saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude solid compound 1l (95 mg). The product was used directly in the next reaction without purification.

[0172] MS m / z(ESI): 614.2 [M+1].

[0173] Step 9

[0174] (5S,5aS,6S,9R)-2-chloro-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylbridgednaphtho[1,8-ab]heptan-14-carboxylic acid tert-butyl

[0175] Ester 1m

[0176] Crude compound 1l (95 mg, 154.7 μmol) and compound 1c (35.5 mg, 234.5 μmol) were dissolved in acetonitrile (4 mL), and anhydrous potassium carbonate (64 mg, 463 μmol) and sodium iodide (70 mg, 467 μmol) were added. The mixture was stirred at 80 °C for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system A to obtain solid compound 1m (80 mg, yield: 81.6%).

[0177] MS m / z(ESI): 633.2 [M+1].

[0178] Step 10 (5S,5aS,6S,9R)-2-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophene-4-yl)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylbridged naphtho[1,8-ab]heptacyclo-14-carboxylic acid tert-butyl ester 1o

[0179] Compound 1m (20 mg, 31.6 μmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoborhexacyclohex-2-yl)-7-fluorobenzo[b]thiophene-2-yl) tert-butyl carbamate 1n (19 mg, 47 μmol, prepared by the method disclosed in preparation 23 on page 39 of patent application "WO2022261154A1"), tetrakis(triphenylphosphine)palladium (7 mg, 6.1 μmol), and cesium carbonate (30 mg, 92 μmol) were mixed with N,N-dimethylformamide (1 mL), purged with nitrogen, and reacted at 90 °C for 2 hours. After the reaction solution was cooled to room temperature, it was diluted with ethyl acetate, dried with anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure to obtain crude solid compound 1o (15 mg). The product was used directly in the next reaction without purification.

[0180] MS m / z(ESI): 889.2 [M+1].

[0181] Step 11

[0182] 2-Amino-7-fluoro-4-((5S,5aS,6S,9R)-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylbridgednaphtho[1,8-ab]heptan-2-yl)benzo

[0183] [b]Thiophene-3-carboxynitrile 1

[0184] The crude compound 1O (15 mg, 16.9 μmol) was dissolved in acetonitrile (1 mL), and 0.5 mL of 4 M 1,4-dioxane hydrochloric acid solution was added under ice bath conditions. After reacting for 0.5 hours, the reaction was continued at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by preparative high-performance liquid chromatography (Waters-2545, column: Da Cao ODS-BIO C18, 30*150 mm, 5 μm; mobile phase: aqueous phase (10 mmol / L ammonium bicarbonate) and acetonitrile, gradient ratio: acetonitrile 54%-69% for 15 min, flow rate: 30 mL / min, column temperature: room temperature) to obtain solid compound 1 (4 mg, yield: 34.4%).

[0185] MS m / z(ESI): 687.3 [M-1].

[0186] 1 H NMR (500MHz, CD3OD): δ7.39(dd,1H),7.04(t,1H),6.51(d,1H),5.41(dd,1H),4.5–4.46(m,2H),4.40(d,1H),4.10(d,1H),3.7 0(d,1H),3.60(d,1H),3.19(s,1H),2.62–2.31(m,8H),2.09(d,3H),1.92–1.75(m,3H),1.61(d,3H),0.75(s,2H),0.52(s,2H).

[0187] Test Example 1: Biological Evaluation of GP2d and AGS Cell 3D Proliferation Inhibition Experiment

[0188] Experimental methods

[0189] GP2d cells were cultured in complete medium, namely DMEM / high glucose medium (Hyclone, SH30243.01) containing 10% fetal bovine serum (Corning, 35-076-CV). On the first day of the experiment, GP2d cells were seeded at a density of 1000 cells / well in 96-well low-absorption plates (Corning, CLS7007-24EA) using complete medium, with 90 μL of cell suspension per well. After centrifugation at 2000 rpm for 5 minutes at room temperature, the cells were incubated overnight at 37°C in a 5% CO2 cell culture incubator.

[0190] AGS cells were cultured in complete medium, namely RPMI 1640 medium (Hyclone, SH30809.01) containing 10% fetal bovine serum (Corning, 35-076-CV). On the first day of the experiment, AGS cells were seeded at a density of 1000 cells / well in 96-well low-absorption plates (Corning, CLS7007-24EA) using complete medium, with 90 μL of cell suspension per well. After centrifugation at 2000 rpm for 5 minutes at room temperature, the cells were incubated overnight at 37°C in a 5% CO2 cell culture incubator.

[0191] On the second day, 10 μL of serially diluted test compounds prepared with complete culture medium was added to each well. For GP2d cells, the final concentrations were determined by nine 5-fold serial dilutions starting from 1 μM; for AGS cells, the final concentrations were determined by nine 5-fold serial dilutions starting from 10 μM. A blank control containing 0.5% DMSO was included in both wells. The plates were incubated at 37°C in a 5% CO2 incubator for 5 days. On the seventh day, the 96-well cell culture plate was removed, and 50 μL of the test compound was added to each well. The 3D Cell Viability Assay reagent (Promega, G9682) was shaken at room temperature in the dark for 25 minutes, then mixed by pipetting and aspiration. 100 μL of the mixture was taken from each well and transferred to a white, opaque 96-well plate (PerkinElmer, 6005290). The luminescence signal value was read using a multi-functional microplate reader (PerkinElmer, EnVision2105).

[0192] The IC50 of the compound's inhibitory activity was calculated using Graphpad Prism software. 50 The values ​​are shown in Table 1 below.

[0193] Table 1. Data on the inhibitory activity of AGS and GP2d cells on 3D proliferation.

[0194]

[0195] Conclusion: The compound disclosed herein has a good inhibitory effect on the 3D proliferation of AGS and GP2d cells.

[0196] Test Example 3: Biological Evaluation of AsPC-1 Cell 3D Proliferation Inhibition Experiment

[0197] On day 1, AsPC-1 cells that had grown well and reached 70%-80% confluence were digested and resuspended in RPMI 1640 (Hyclone, SH30809.01) medium containing 10% FBS, and the cell density was adjusted to the desired level. 90 μL of cell suspension was added to each well of a U-shaped, low-adsorption 96-well plate (Corning, CLS7007-24EA), resulting in a cell density of 1500 cells / well. The cell plates were centrifuged at 2500 rpm for 5 minutes and then incubated overnight at 37°C in a 5% CO2 incubator. On day 2, the 20 mM test compound dissolved in DMSO was diluted to an initial concentration of 2 mM with DMSO, and then serially diluted 5-fold for a total of 9 concentration points. The control wells contained DMSO. The serially diluted compounds were then further diluted 20-fold with medium. 10 μL of the diluted test compound was added to each well of the cell plate, resulting in a final concentration of 10 μM, representing the 9 concentration points of the 5-fold serial dilutions. Cell culture wells containing 0.5% DMSO were set up as solvent control wells, and wells containing only culture medium and 0.5% DMSO were set as blank control wells. Each compound concentration and control well was replicated, with a final DMSO concentration of 0.5% in each well. The cell culture plates were centrifuged at 2500 rpm for 3 minutes and then incubated at 37°C in a 5% CO2 incubator for 5 days. On the seventh day, the 96-well cell culture plates were removed, and 50 μL of luminescent cell viability assay reagent was added to each well. In the 3D Cell Viability Assay (Promega, G9683), after shaking at room temperature in the dark for 25 minutes, mix thoroughly by pipetting up and down, then transfer 100 μL to each well into a white, opaque OptiPlate. TM -96-well plates (PerkinElmer, 6005290) were used to read the luminescence signal values ​​using a multi-functional microplate reader (PerkinElmer, EnVision2105).

[0198] Calculate the inhibition rate using the following formula: Inhibition rate = (Luminous value) / (Emitting value) 溶媒对照孔 -Luminescence value 受试化合物 ) / (luminous value) 溶媒对照孔 -Luminescence value 空白对照孔 ()×100%. Using GraphPadPrism software, curves were plotted based on the concentrations of the compounds and their corresponding inhibition rates, and the IC50 of the compounds was calculated. 50 value.

[0199] Table 2. Data on the 3D proliferation inhibition activity of AsPC-1 cells

[0200] Example number <![CDATA[AsPC-1 / IC 50 (nM)]]> 1 2.4

[0201] Conclusion: The compound disclosed herein has a good inhibitory effect on the 3D proliferation of AsPC-1 cells.

[0202] Test Example 4: Pharmacokinetic Evaluation

[0203] 1. Abstract

[0204] Using balb / c nude mice as test animals, the drug concentrations in plasma at different times after oral administration (i.g.) of the compound of the example to balb / c nude mice were determined by LC / MS / MS method. The pharmacokinetic behavior of the compound of the present disclosure in balb / c nude mice was studied to evaluate its pharmacokinetic characteristics.

[0205] 2. Test Protocol

[0206] 2.1 Test Drug

[0207] Compound 1.

[0208] 2.2 Test Animals

[0209] Nine balb / c nude mice, female, provided by Vital River Laboratory Animal Technology Co., Ltd., production license SCXK(Zhe)2019-0001.

[0210] 2.3 Drug Preparation

[0211] Weighed a certain amount of the test compound respectively, added 5% DMF + 45% PG + 50% (10% HS15-pH7.4 buffer) + 400mpk SNAC, and prepared a 4mg / mL colorless and clear solution.

[0212] 2.4 Drug Administration

[0213] The administration dose was 40.0mg / kg, and the administration volume was 10mL / kg.

[0214] 3. Operations

[0215] Before drug administration and at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, 24.0 hours after drug administration, 0.1 mL of blood was collected from the orbital cavity, placed in an EDTA-K2 anticoagulant test tube, centrifuged at 10000 rpm for 1 minute (4°C), the plasma was separated within 1 hour, and stored at -20°C for testing. The blood collection to centrifugation process was operated under ice bath conditions.

[0216] Determined the content of the test compound in the plasma of balb / c nude mice after administration of drugs at different concentrations: Took 25 μL of the plasma samples of balb / c nude mice at each time after administration, added 200 μL of acetonitrile containing internal standard (Compound 1: the internal standard was verapamil 100 ng / mL), vortex mixed for 5 minutes, and centrifuged at 3700 rpm for 20 minutes. The supernatant was mixed with water (1:1). Took 0.5 - 2 μL of the supernatant for LC / MS / MS analysis.

[0217] 4. Pharmacokinetic Parameter Results

[0218] Table 3. Pharmacokinetic parameters of the compounds disclosed herein in Balb / c nude mice.

[0219]

[0220] Conclusion: The compound disclosed herein exhibits favorable pharmacokinetic properties in balb / c nude mice.

[0221] Example 2 Preparation of hydrochloride

[0222] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2M hydrochloric acid-ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous hydrochloride salt, and the XRPD spectrum showed no obvious characteristic peaks. Ionic analysis revealed a chloride ion content of 8.29%.

[0223] Example 3 Preparation of hydrochloride

[0224] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of the solvent in Table 4, and 6 μL of 2M hydrochloric acid-ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous hydrochloride salt.

[0225] Table 4

[0226]

[0227]

[0228] Example 4: Preparation of Sulfate

[0229] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2M sulfuric acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous sulfate, with no obvious characteristic peaks in the XRPD spectrum. Ionic results showed that the sulfate content was 12.62%.

[0230] Example 5: Preparation of Sulfate

[0231] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of the solvent in Table 5, and 6 μL of 2M sulfuric acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous sulfate.

[0232] Table 5

[0233] solvent Crystal form acetone Sulfate amorphous 2-Methyltetrahydrofuran Sulfate amorphous

[0234] Example 6 Preparation of Sulfate Crystal Form I

[0235] 7 mg of the amorphous compound shown in Formula 1 was added to 0.15 mL of 10% water / methanol and 10.7 μL of 2M sulfuric acid ethanol solution. The mixture was stirred to precipitate, centrifuged, and the solid was dried under vacuum to obtain the solid product. X-ray powder diffraction analysis identified this product as sulfate crystal form I. The XRPD spectrum is shown below. Figure 1 The positions of its characteristic peaks are shown in Table 6.

[0236] Table 6

[0237]

[0238] Example 7 Preparation of Phosphate

[0239] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2 M phosphate ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous phosphate, with no obvious characteristic peaks in the XRPD spectrum. Ionic analysis revealed a phosphate content of 15.03%.

[0240] Example 8 Preparation of Phosphate

[0241] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of solvent from Table 7, and 6 μL of 2M phosphate ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous phosphate.

[0242] Table 7

[0243] solvent Crystal form acetone Phosphate amorphous 2-Methyltetrahydrofuran Phosphate amorphous

[0244] Example 9 Preparation of Phosphate Crystal Form I

[0245] 7 mg of the amorphous compound shown in Formula 1 was added to 0.15 mL of 10% water / methanol and 10.7 μL of 2M phosphate ethanol solution. The mixture was stirred to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was phosphate crystal form I, and the XRPD spectrum is shown below. Figure 2 The positions of its characteristic peaks are shown in Table 8. The DSC spectrum shows an endothermic peak at 196.2℃. The TGA spectrum shows a weight loss of 2.02% between 33℃ and 143℃. Ion results show a phosphate content of 16.85%.

[0246] Table 8

[0247]

[0248] Example 10 Preparation of Methanesulfonate

[0249] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2M methanesulfonic acid in ethanol was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous methanesulfonate, with no obvious characteristic peaks in the XRPD spectrum. Ionic analysis revealed a methanesulfonic acid content of 20.96%.

[0250] Example 11 Preparation of methanesulfonate

[0251] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of solvent from Table 9, and 6 μL of 2M methanesulfonic acid in ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous methanesulfonate.

[0252] Table 9

[0253] solvent Crystal form acetone amorphous methanesulfonate 2-Methyltetrahydrofuran amorphous methanesulfonate

[0254] Example 12 Preparation of acetate

[0255] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of Table 10 solvent, 6 μL of 2M acetic acid-ethanol solution and 0.1 mL of n-heptane were added, and the mixture was stirred at room temperature to precipitate. After centrifugation, the solid was dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous acetate, and the XRPD spectrum showed no obvious characteristic peaks.

[0256] Table 10

[0257]

[0258]

[0259] Example 13 Preparation of citrate

[0260] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2 M citric acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous citrate salt, and the XRPD spectrum showed no obvious characteristic peaks. NMR results showed that the salt ratio of the compound to citric acid was 1:1.

[0261] Example 14 Preparation of Citrate

[0262] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of solvent from Table 11, and 6 μL of 2M citric acid-ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous citrate salt.

[0263] Table 11

[0264] solvent Crystal form acetone amorphous citrate 2-Methyltetrahydrofuran amorphous citrate

[0265] Example 15 Preparation of Succinate

[0266] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 1.4 mg of succinic acid was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous succinate, and the XRPD spectrum showed no obvious characteristic peaks. Ionic analysis revealed that the succinic acid content was 17.52%.

[0267] Example 16 Preparation of Succinate

[0268] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of solvent from Table 12, and 1.4 mg of succinic acid was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous succinate.

[0269] Table 12

[0270] solvent Crystal form acetone Succinate amorphous 2-Methyltetrahydrofuran Succinate amorphous

[0271] Example 17 Preparation of p-Toluenesulfonate

[0272] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2 M p-toluenesulfonic acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous p-toluenesulfonate, and the XRPD spectrum showed no obvious characteristic peaks. Ionic analysis revealed that the p-toluenesulfonic acid content was 30.49%.

[0273] Example 18 Preparation of p-Toluenesulfonate

[0274] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of solvent from Table 13, and 6 μL of 2 M p-toluenesulfonic acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous p-toluenesulfonate.

[0275] Table 13

[0276] solvent Crystal form acetone p-Toluenesulfonate amorphous 2-Methyltetrahydrofuran p-Toluenesulfonate amorphous

[0277] Example 19 Preparation of maleate

[0278] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2 M maleic acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous maleate salt, and the XRPD spectrum showed no obvious characteristic peaks. Ionic analysis revealed a maleic acid content of 16.94%.

[0279] Example 20 Preparation of maleate

[0280] Dissolve 7 mg of the compound shown in Formula 1 in 0.1 mL of the solvent in Table 14, add 6 μL of 2 M maleic acid ethanol solution, stir at room temperature to precipitate, centrifuge, and dry the solid under vacuum to obtain the solid product.

[0281] Table 14

[0282] solvent Crystal form acetone Maleate amorphous 2-Methyltetrahydrofuran Maleate amorphous

[0283] Example 21 Preparation of fumarate crystal form I

[0284] Dissolve 7 mg of the compound shown in Formula 1 in 0.2 mL of ethyl acetate, add 1.3 mg of fumaric acid, stir at room temperature to precipitate, centrifuge, and dry the solid under vacuum to obtain the solid product.

[0285] X-ray powder diffraction analysis identified the product as fumarate crystal form I, and the XRPD spectrum is shown below. Figure 3 The positions of its characteristic peaks are shown in Table 15. Ion results show that the fumaric acid content is 14.23%.

[0286] Table 15

[0287]

[0288] Example 22 Preparation of fumarate crystal form I

[0289] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of solvent from Table 16, and 1.3 mg of fumaric acid was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was fumarate crystal form I.

[0290] Table 16

[0291] solvent Crystal form acetone Fumarate crystal form I 2-Methyltetrahydrofuran Fumarate crystal form I

[0292] Example 23 Preparation of L-malate

[0293] 7 mg of the compound shown in Formula 1 was dissolved in the solvent in Table 17, and 6 μL of 2M L-malic acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous L-malate, with no obvious characteristic peaks in the XRPD spectrum. Ionic results showed that the L-malic acid content was 28.11%.

[0294] Table 17

[0295] solvent Crystal form 0.2 mL ethyl acetate L-malate amorphous 0.1 mL acetone L-malate amorphous 0.1 mL 2-Methyltetrahydrofuran L-malate amorphous

[0296] Example 24 Preparation of L-malate crystal form I

[0297] 7 mg of the compound shown in Formula 1 was added to 0.1 mL of ethyl acetate and 10.7 μL of 2 M L-malic acid ethanol solution. The mixture was stirred to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as L-malate crystal form I. The XRPD spectrum is shown below. Figure 4 The positions of its characteristic peaks are shown in Table 18. The DSC spectrum shows that the endothermic peak has a peak value of 100.17℃.

[0298] TGA spectra showed a weight loss of 4.74% between 33℃ and 147℃. Ion analysis revealed an L-malic acid content of 20.65%. Table 18

[0299]

[0300] Example 25: Preparation of L-malate crystal form I

[0301] Add 7 mg of the compound shown in Formula 1 to 0.1 mL of acetone and 10.7 μL of 2 M L-malic acid ethanol solution, stir to precipitate, centrifuge, and dry the solid under vacuum to obtain the solid product.

[0302] Example 26 Preparation of L-Tartrate Crystal Form I

[0303] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2 M L-tartaric acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as L-tartarate crystal form I, and the XRPD spectrum is shown below. Figure 5 The positions of its characteristic peaks are shown in Table 19. The DSC spectrum shows endothermic peaks at 102.17℃ and 117.01℃. The TGA spectrum shows a weight loss of 5.88% between 32℃ and 175℃. Ion analysis indicates an L-tartaric acid content of 16.8%.

[0304] Table 19

[0305]

[0306] Example 27 Preparation of L-Tartrate Crystal Form I

[0307] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of Table 20 solvent, and 6 μL of 2 M L-tartaric acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified the product as L-tartaric acid crystal form I.

[0308] Table 20

[0309] solvent Crystal form acetone L-Tartrate Crystal Form I 2-Methyltetrahydrofuran L-Tartrate Crystal Form I

[0310] Example 28 Preparation of L-Tartrate Crystal Form I

[0311] 7 mg of the amorphous compound shown in Formula 1 was added to 0.15 mL of 10% water / methanol and 10.67 μL of 2 M L-tartaric acid ethanol solution. The mixture was stirred to precipitate, centrifuged, and dried under vacuum to obtain the solid product.

[0312] Example 29: Preparation of L-Tartrate Crystal Form II

[0313] 7 mg of the amorphous compound shown in Formula 1 was added to 0.15 mL of acetone and 10.67 μL of 2 M L-tartaric acid ethanol solution. The mixture was stirred to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as L-tartarate crystal form II. The XRPD spectrum is shown below. Figure 6 The positions of its characteristic peaks are shown in Table 21. The DSC spectrum shows endothermic peaks at 105.17℃ and 158.01℃. The TGA spectrum shows a weight loss of 9.11% between 35℃ and 189℃. Ionic results show an L-tartaric acid content of 14.97%.

[0314] Table 21

[0315]

[0316] Example 30 Preparation of glutarate crystal form I

[0317] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 1.5 mg of glutaric acid was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as glutarate crystal form I. The XRPD spectrum is shown below. Figure 7 The positions of its characteristic peaks are shown in Table 22. The DSC spectrum shows an endothermic peak at 99.16℃. The TGA spectrum shows a weight loss of 3.89% from 32℃ to 164℃. Ion results show a glutaric acid content of 16.78%.

[0318] Table 22

[0319]

[0320]

[0321] Example 31 Preparation of glutarate crystal form I

[0322] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of solvent from Table 23, and 1.5 mg of glutaric acid was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified the product as glutarate crystal form I.

[0323] Table 23

[0324] solvent Crystal form acetone Glutarate crystal form I 2-Methyltetrahydrofuran Glutarate crystal form I

[0325] Example 32 Preparation of glutarate crystal form II

[0326] 7 mg of the amorphous compound shown in Formula 1 was added to 0.1 mL of ethyl acetate and 2.8 mg of glutaric acid. The mixture was stirred to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as glutarate crystal form II. The XRPD spectrum is shown below. Figure 8 The positions of its characteristic peaks are shown in Table 24. The DSC spectrum shows an endothermic peak at 111.16℃. The TGA spectrum shows a weight loss of 5.63% from 32℃ to 139℃. Ion results show a glutaric acid content of 26%.

[0327] Table 24

[0328]

[0329] Example 33 Preparation of Benzoate Crystal Form I

[0330] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2M benzoic acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as benzoate crystal form I. The XRPD spectrum is shown below. Figure 9 The positions of its characteristic peaks are shown in Table 25. The DSC spectrum shows an endothermic peak at 169℃. The TGA spectrum shows a weight loss of 1.95% from 32℃ to 150℃. Ion analysis indicates a benzoic acid content of 14.93%.

[0331] Table 25

[0332]

[0333] Example 34 Preparation of Benzoate Crystal Form I

[0334] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of Table 26 solvent, and 6 μL of 2M benzoic acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was benzoate crystal form I.

[0335] Table 26

[0336] solvent Crystal form acetone Benzoate crystal form I 2-Methyltetrahydrofuran Benzoate crystal form I

[0337] Example 35 Preparation of o-benzoylsulfonylimide complex

[0338] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 2.1 mg of o-benzoylsulfonylimide was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified the product as an amorphous o-benzoylsulfonylimide complex, with no obvious characteristic peaks in the XRPD spectrum.

[0339] Example 36 Preparation of o-benzoylsulfonylimide complex

[0340] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of 2-methyltetrahydrofuran, and 2.1 mg of o-benzoylsulfonylimide was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous o-benzoylsulfonylimide complex.

[0341] Example 37 Preparation of o-benzoylsulfonylimide complex crystal form I

[0342] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of acetone, and 2.1 mg of o-benzoylsulfonylimide was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as o-benzoylsulfonylimide complex crystal form I. The XRPD spectrum is shown below. Figure 10 The positions of its characteristic peaks are shown in Table 27. The DSC spectrum shows endothermic peaks at 68.18℃, 106.18℃, and 160.02℃. The TGA spectrum shows a weight loss of 3.92% between 31℃ and 115℃. NMR results show that the compound is in a 1:1 ratio with o-benzoylsulfonylimide.

[0343] Table 27

[0344]

[0345]

[0346] Example 38 Preparation of o-benzoylsulfonylimide complex crystal form I

[0347] 7 mg of the amorphous compound shown in Formula 1 was added to 0.15 mL of 10% water / methanol and 4.2 mg of o-benzoylsulfonylimide. The mixture was stirred at room temperature to precipitate the product. After centrifugation, the solid was dried under vacuum to obtain the solid product.

[0348] Example 39 Preparation of Arginine Complex Crystal Form a

[0349] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of acetone, and 1.9 mg of arginine was added. The mixture was stirred at room temperature to precipitate, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as arginine complex crystal form a, and the XRPD spectrum is shown below. Figure 11 The positions of its characteristic peaks are shown in Table 28. The DSC spectrum shows endothermic peaks at 68.18℃, 106.18℃, and 160.02℃. The TGA spectrum shows a weight loss of 3.92% between 31℃ and 115℃.

[0350] Table 28

[0351]

[0352] Example 40: Factors Affecting Stability

[0353] The aforementioned o-benzoylsulfonylimide complex crystal form I was laid out in an open position, and the stability of the samples was investigated under light (4500 Lux), high temperature (40℃, 60℃), and high humidity (RH 75%, RH 92.5%) conditions. The sampling period was 30 days.

[0354] Conclusion: Experiments on influencing factors show that the physicochemical stability of the crystal form disclosed in this study is good under high temperature (40℃ and 60℃) and high humidity (75% and 92.5%) conditions for 30 days.

[0355] Example 41: Long-term accelerated test stability

[0356] The stability of the aforementioned o-benzoylsulfonylimide complex crystal form I was investigated under conditions of 25℃ / 60%RH and 40℃ / 75%RH.

[0357] Conclusion: Long-term accelerated experiments show that the physicochemical stability of the disclosed crystal form is good under conditions of 25℃ / 60%RH and 40℃ / 75%RH for one month.

Claims

1. A 2-amino-7-fluoro-4-((5-)-represented by Formula 1 S 5 aS 6 S 9 R )-1-fluoro-12-((1-((4-(fluoromethylidene)piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methyl-5 a ,6,7,8,9,10-hexahydro-5 H -4-oxa-3,10 a ,11,13,14-pentazaza-6,9-methylnaphtho[1,8- ab [[heptan-2-yl)benzo[]] b Pharmaceutically acceptable salts of thiophene-3-carboxynitrile, wherein the pharmaceutically acceptable salts are selected from hydrochloride, sulfate, phosphate, methanesulfonate, acetate, citrate, succinate, p-toluenesulfonate, maleate, fumarate, L-malate, L-tartrate, glutarate, and benzoate. 。 2. The sulfate crystal form I of the compound of formula 1 according to claim 1, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 4.924, 9.872, 17.934, and 21.

926.

3. The sulfate crystal form I according to claim 2, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 1.

4. A method for preparing sulfate crystal form I as described in claim 2 or 3, the method comprising adding the compound of formula 1 to 10% water / methanol, then adding sulfuric acid ethanol solution, and stirring.

5. The L-tartrate crystal form II of the compound of Formula 1 according to claim 1, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 5.073, 8.094, 10.179, 19.316, and 20.

966.

6. The L-tartrate crystal form II according to claim 5, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 6.

7. A method for preparing L-tartrate crystal form II as described in claim 5 or 6, the method comprising adding the compound of formula 1 to acetone, then adding an L-tartrate ethanol solution, and stirring.

8. The crystal form according to any one of claims 2-3 and 5-6, wherein the 2 θ The angular error range is ±0.

20.

9. A pharmaceutical composition comprising a pharmaceutically acceptable salt of a compound of formula 1 as claimed in claim 1, or a crystal form as claimed in any one of claims 2-3, 5-6, and optionally a pharmaceutically acceptable excipient.

10. A method for preparing a pharmaceutical composition, comprising the following steps: The step of mixing a pharmaceutically acceptable salt of the compound of formula 1 as claimed in claim 1, or the crystal form as claimed in any one of claims 2-3, 5-6, with a pharmaceutically acceptable excipient.

11. Use of a pharmaceutically acceptable salt of the compound of formula 1 according to claim 1, or the crystal form according to any one of claims 2-3, 5-6, or the pharmaceutical composition according to claim 9 in the preparation of a KRAS inhibitor.

12. Use of a pharmaceutically acceptable salt of the compound of formula 1 as claimed in claim 1, or the crystal form as claimed in any one of claims 2-3, 5-6, or the pharmaceutical composition as claimed in claim 9, as a KRAS inhibitor in the preparation of a medicament for the treatment and / or prevention of cancer.

Images