Medicinal salt of benzothiophene substituted fused ring compound, crystal form and application thereof

Abstract

The invention relates to a pharmaceutically acceptable salt of a benzothiophene substituted fused ring compound, and a crystal form and application thereof. Specifically, the present disclosure provides a pharmaceutically acceptable salt of 2-amino-7-fluoro-4-((5S, 5aS, 6S, 9R)-1-fluoro-12-((1-((4-(fluoromethylsubunit) piperidine-1-yl) methyl) cyclopropyl) methoxyl)-5-methyl-5a, 6, 7, 8, 9, 10-hexahydro-5H-4-oxa-3, 10a, 11, 13, 14-pentaaza-6, 9-methylbridged naphtho [1, 8-ab] heptano-2-yl) benzo [b] thiophene-3-formonitrile, a crystal form and a preparation method thereof, and a preparation method of the pharmaceutically acceptable salt, the crystal form and the preparation method thereof. Corresponding salts have good stability and can be better used for clinical treatment. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to a pharmaceutically acceptable salt of a benzothiophene-substituted condensed ring compound, a crystal form thereof and uses thereof. Background Art

[0002] RAS is one of the oncogenes with the highest mutation rate in tumors, and about 30% of human malignant tumors are related to mutations in the RAS gene. The RAS family includes KRAS, NRAS, and HRAS, among which KRAS mutations are the most common, accounting for about 85%. KRAS mutations are common in solid tumors, and high-frequency mutations exist in the three major fatal human cancers: lung cancer (17%), colorectal cancer (33%), and pancreatic cancer (61%). Among the KRAS gene mutations, 97% are mutations in the 12th or 13th amino acid residues, among which G12D is an important mutation. Data analysis of European and American populations shows that in pancreatic cancer, colorectal cancer, and non-small cell lung cancer, G12D mutations account for 36%, 12%, and 4% of patients, respectively.

[0003] After KRAS is activated, it regulates cell proliferation, survival, migration, metabolism and other functions through numerous downstream signaling pathways represented by RAF-MEK-ERK, PI3K-AKT-mTOR and TIAM1-RAc. After the KRAS gene mutates, the protein remains in an activated state, resulting in the continuous activation of downstream signaling pathways and promoting tumorigenesis.

[0004] PCT / CN2023 / 109598 provides a KRAS G12D inhibitor, whose chemical name is 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-methylnaphtho[1,8-ab]heptylcyclo-2-yl)benzo[b]thiophene-3-carbonitrile, having a structure shown in Formula 1,

[0005]

[0006] Salt formation can improve some undesirable physicochemical or biological properties of drugs. It is of great significance to develop salts with better physicochemical or pharmaceutical properties than 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-methano[1,8-ab]heptylcyclo-2-yl)benzo[b]thiophene-3-carbonitrile. In view of the importance of solid drug crystal forms and their stability in clinical treatment, in-depth research on the polymorphic forms of pharmaceutically acceptable salts 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-pentaaza-6,9-methanaphtho[1,8-ab]heptylcyclo-2-yl)benzo[b]thiophene-3-carbonitrile is also of great significance for the development of drugs suitable for industrial production and with good biological activity. Summary of the invention

[0007] The present disclosure provides a pharmaceutically acceptable salt of a compound of formula 1, 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-methylnaphtho[1,8-ab]heptyl-2-yl)benzo[b]thiophene-3-carbonitrile, 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] The present 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, wherein the acid is 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 solvent used for salt formation in the present invention is selected from but not limited to acetone, ethyl acetate, 2-methyltetrahydrofuran, methanol, and 10% water / methanol.

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

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

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

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

[0015] Furthermore, in an optional embodiment, the method for preparing the aforementioned pharmaceutically acceptable salt further comprises the steps of crystallization, filtering, washing or drying.

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

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

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

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

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

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

[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 1:1-1:2.

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

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

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

[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 1:1-1:2.

[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 represented by Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peak at a diffraction angle 2θ in the range of 3-50°.

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

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

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

[0035] The present disclosure also provides a sulfate crystal form I of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 4.924, 9.872, 17.934, and 21.926.

[0036] In some embodiments, the sulfate crystalline form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 1 shown.

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

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

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

[0040] The present disclosure also provides a phosphate crystal form I of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 5.572 and 9.488.

[0041] In some embodiments, the phosphate crystal form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 2 shown.

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

[0043] In some embodiments, the mesylate salt of the compound represented by Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peak at a diffraction angle 2θ in the range of 3-50°.

[0044] The present disclosure also provides a method for preparing an amorphous methanesulfonate salt of a compound represented by Formula 1, comprising the steps of dissolving the compound represented by Formula 1 in a solvent A, adding a methanesulfonic acid ethanol solution, and stirring.

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

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

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

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

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

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

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

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

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

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

[0055] The fumarate crystalline form I of the compound represented by Formula 1 provided in the present disclosure has characteristic peaks at 9.184, 15.751 and 19.708 in an X-ray powder diffraction pattern represented by a diffraction angle 2θ.

[0056] In some embodiments, the fumarate salt form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 3 shown.

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

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

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

[0060] The present disclosure also provides a L-malate crystalline form I of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 9.680, 14.709, 15.093, 15.515, and 19.891.

[0061] In some embodiments, the L-malate crystalline form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 4 shown.

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

[0063] The L-tartrate crystalline form I of the compound represented by Formula 1 provided in the present disclosure has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, and has characteristic peaks at 4.959, 9.606, 15.022, and 19.631.

[0064] In some embodiments, the L-tartrate salt form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 5 shown.

[0065] The present disclosure also provides a method for preparing L-tartrate crystalline form I of the compound represented by Formula 1, the method comprising the steps of dissolving the compound represented by Formula 1 in solvent A, adding L-tartaric acid ethanol solution, and stirring.

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

[0067] The present disclosure also provides a L-tartrate salt form II of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by 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 L-tartrate salt form II of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 6 shown.

[0069] The present disclosure also provides a method for preparing L-tartrate crystal form II of the compound represented by Formula 1, the method comprising the steps of adding the compound represented by Formula 1 to acetone, then adding L-tartaric acid ethanol solution, and stirring.

[0070] The glutaric acid salt form I of the compound represented by Formula 1 provided in the present disclosure has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, and has characteristic peaks at 10.374, 15.175, and 19.746.

[0071] In some embodiments, the glutaric acid salt form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 7 shown.

[0072] The present disclosure also provides a method for preparing glutaric acid salt crystal form I of the compound represented by Formula 1, the method comprising the steps of dissolving the compound represented by Formula 1 in solvent A, adding glutaric acid, and stirring.

[0073] The present disclosure also provides a glutaric acid salt crystal form II of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 8.490, 13.020, 15.362, 18.433, and 21.619.

[0074] In some embodiments, the glutaric acid salt form II of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 8 shown.

[0075] The present disclosure also provides a method for preparing glutaric acid salt crystal form II of the compound represented by formula 1, the method comprising the steps of adding the compound represented by formula 1 to ethyl acetate, then adding glutaric acid, and stirring.

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

[0077] In some embodiments, the benzoate salt form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Fig. 9 shown.

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

[0079] The present disclosure also provides a complex formed by the compound represented by Formula 1 and a ligand, wherein the ligand is selected from arginine and o-benzoylsulfonyl imide.

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

[0081] The present disclosure provides a method for preparing an amorphous o-benzoylsulfonyl imide complex of a compound represented by Formula 1. The method comprises the steps of dissolving the compound represented by Formula 1 in a solvent D, adding o-benzoylsulfonyl imide, and stirring.

[0082] The present disclosure also provides a crystalline form I of an o-benzoylsulfonyl imide complex of a compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ, having characteristic peaks at 6.985, 8.161, 11.381, 16.472, 19.186, 23.303, and 29.367.

[0083] In some embodiments, the o-benzoylsulfonyl imide complex crystalline form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, and 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 o-benzoylsulfonyl imide complex crystalline form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, and 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 o-benzoylsulfonyl imide complex crystalline form I of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Fig.10 shown.

[0086] The present disclosure also provides a method for preparing the o-benzoylsulfonyl imide complex crystalline form I of the compound of formula 1, the method comprising the steps of adding the compound of formula 1 to acetone or 10% water / methanol, then adding o-benzoylsulfonyl imide, and stirring.

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

[0088] In some embodiments, the arginine complex crystalline form a of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Fig.11 shown.

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

[0090] The present disclosure also provides a pharmaceutical composition, which contains 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-benzoylsulfonyl imide complex crystal form I or arginine complex crystal form a, and pharmaceutical excipients selected from pharmaceutically acceptable excipients.

[0091] The present disclosure also provides a pharmaceutical composition, which is prepared from 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-benzoylsulfonyl imide complex crystal form I or arginine complex crystal form a, and optionally a pharmaceutically acceptable excipient.

[0092] The present disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing 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-benzoylsulfonyl imide complex crystal form I or arginine complex crystal form a with a pharmaceutically acceptable excipient.

[0093] The present disclosure also provides 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-benzoylsulfonyl imide complex crystal form I or arginine complex crystal form a or the aforementioned composition in the preparation of a method for preventing and / or treating cancer.

[0094] The use disclosed herein, 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 cancer, oral cancer, thyroid cancer, skin cancer, 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 the present disclosure refers to the diffraction angle, θ is the Bragg angle, and the unit is ° or degree; the error range of each characteristic peak 2θ is ±0.20 (including the case where the number exceeding 1 decimal place is rounded off), 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 the present disclosure, such as the data of the measured and calculated content of the relevant substances, inevitably have a certain degree of error. Generally speaking, ±10% is within the reasonable error range. There is a certain degree of error variation depending on the context where it is used, and the error variation does not exceed ±10%, and can be ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2% or ±1%, preferably ±5%.

[0097] The starting material used in the crystal preparation method of the present invention can be a compound in any form, and specific forms include but are not limited to: amorphous, any crystal form, hydrate, solvate, etc.

[0098] The drying temperature in the present disclosure is generally 25° C.-100° C., preferably 40° C.-70° C., and the drying can be performed under normal pressure or reduced pressure.

[0099] The crystallization methods described in the present disclosure include room temperature crystallization, cooling crystallization, volatile solvent crystallization, adding seed crystals to induce crystallization, etc. The cooling temperature is selected from below 65°C, preferably -10°C to 60°C, and stirring can also be performed during the crystallization process.

[0100] The "differential scanning calorimetry or DSC" described in the present disclosure refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining a constant temperature of the sample to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

[0101] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "9103 Drug Hygroscopicity Guidelines" in the fourth volume of the 2015 edition of the Chinese Pharmacopoeia,

[0102] Deliquesce: Absorbs enough water to form a liquid;

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

[0104] Hygroscopic: weight gain due to moisture absorption is less than 15% but not less than 2%;

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

[0106] No or almost no hygroscopicity: weight gain due to moisture is less than 0.2%.

[0107] The "excipients" described in the present disclosure include, but are not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent or emulsifier approved by the U.S. Food and Drug Administration for use by humans or livestock animals. BRIEF DESCRIPTION OF THE DRAWINGS

[0108] Figure 1 This is the XRPD spectrum of compound 1 sulfate salt form I.

[0109] Figure 2 This is the XRPD spectrum of Compound 1 phosphate form I.

[0110] Figure 3 This is the XRPD spectrum of compound 1 fumarate salt form I.

[0111] Figure 4 This is the XRPD spectrum of Compound 1 L-malate Form I.

[0112] Figure 5 This is the XRPD spectrum of Compound 1 L-tartrate Form I.

[0113] Figure 6 This is the XRPD spectrum of compound 1L-tartrate salt form II.

[0114] Figure 7 This is the XRPD spectrum of Compound 1 glutaric acid salt Form I.

[0115] Figure 8 This is the XRPD spectrum of Compound 1 glutaric acid salt Form II.

[0116] Fig. 9 This is the XRPD spectrum of Compound 1 benzoate Form I.

[0117] Fig.10 This is the XRPD spectrum of compound 1 o-benzoylsulfonyl imide complex form I.

[0118] Fig.11 This is the XRPD spectrum of compound 1 arginine complex form a. DETAILED DESCRIPTION

[0119] The present disclosure will be explained in more detail below in conjunction with 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 essence and scope of the present disclosure.

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

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

[0122] MS was measured using Agilent 1200 / 1290DAD-6110 / 6120 Quadrupole MS LC / MS (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), Waters ACQuity UPLC-QD / SQD (manufacturer: waters, MS model: waters ACQuity Qda Detector / waters SQ Detector), and THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q Exactive).

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

[0124] High performance liquid chromatography 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 200-300 mesh silica gel as the carrier.

[0126] XRPD is X-ray powder diffraction detection: the measurement is carried out using a BRUKER D8 X-ray diffractometer, and the specific collection information is: Cu anode (40kV, 40mA), Cu-Kα1 ray Kα2 rays Kβ rays Scanning mode: θ / 2θ, scanning range (2θ range): 3°~45°.

[0127] DSC is differential scanning calorimetry: the measurement was performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter, with a heating rate of 10°C / min, the specific temperature range refers to the corresponding spectrum (mostly 25-250°C or 350°C), and a nitrogen purge rate of 50 mL / min.

[0128] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer, with a heating rate of 10°C / min, the specific temperature range refers to the corresponding spectrum (mostly 30-350°C), and a nitrogen purge rate of 50 mL / min.

[0129] DVS stands for dynamic moisture adsorption: SMSDVS Advantage is used for detection. At 25°C, 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 range of humidity is subject to the corresponding graph, and most of the methods listed here are used). The judgment standard is that dm / dt is not greater than 0.002%.

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

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

[0132] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1L.

[0133] The reaction progress in the embodiment is monitored by thin layer chromatography (TLC), the developing solvent used in the reaction, the eluent system of column chromatography used for purifying the compound and the developing solvent system of thin layer chromatography include: A: n-hexane / ethyl acetate system, B: dichloromethane / methanol system, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid can 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-pentaaza-6,9-methano[1,8-ab]heptyl-2-yl)benzo

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

[0137]

[0138] first step

[0139] tert-Butyl 4-(fluoromethylidene)piperidin-1-carboxylate 1b

[0140] 2-((Fluoromethyl)sulfonyl)pyridine (4.2 g, 23.97 mmol) was dissolved in tetrahydrofuran (50 mL), and a 1 M solution of potassium bis(trimethylsilyl)amide in tetrahydrofuran (30 mL) was added at -78°C. The temperature was maintained for reaction for 30 minutes, and then N-tert-butoxycarbonyl-4-piperidone 1a (5 g, 25.09 mmol, Shanghai Shaoyuan) was added. The temperature was maintained for reaction for 3 hours, and then the reaction was restored to room temperature for reaction for 1 hour. Saturated ammonium chloride was added to the reaction solution for quenching, and 3N hydrochloric acid (100 mL) was added. After stirring for 1 hour, it 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 with eluent system B to obtain solid compound 1b (2 g, yield: 37%).

[0141] Step 2

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

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

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

[0145] Step 3

[0146] 2,5,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidin-4-ol 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 reaction was stirred at 110°C for 14 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane, and 20% potassium carbonate solution was added dropwise to adjust the pH to 2-3. After stirring for 2 hours, the mixture was filtered, and the filter cake was washed with water and dried to obtain a 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-hydroxypyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo

[0151] [3.2.1] Octane-8-carboxylic acid tert-butyl ester 1g

[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 the specification of the patent application "WO2022173678A1") was dissolved in tetrahydrofuran (10 mL), and 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, and the reaction was stirred for 2 hours. Water was added to the reaction solution to quench the reaction solution and then concentrated under reduced pressure to obtain a crude solid compound 1g (600 mg). The product was not purified and used directly in the next step.

[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-pentaaza

[0156] -6,9-methylnaphtho[1,8-ab]heptyl-14-carboxylic acid tert-butyl ester 1h

[0157] Compound 1g (78 mg, 159.7 μmol) was dissolved in dichloromethane (2 mL), and N,N-diisopropylethylamine (61.9 mg, 478.9 μmol) and phosphorus oxychloride (122.4 mg, 798.2 μmol) were added under ice bath, and the reaction was stirred for 2 hours. Saturated sodium bicarbonate solution was added to the reaction solution to quench, and dichloromethane (10 mL×2) was used. The organic phases were combined and dried over anhydrous sodium sulfate. The desiccant was removed by filtration and concentrated under reduced pressure to obtain a crude solid compound 1h (75 mg). The product was directly used in the next step without purification.

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

[0159] Step 6

[0160] tert-Butyl (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-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-14-carboxylate

[0161] Ester 1j

[0162] (1-(((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)methanol 1i (1.4 g, 6.4 mmol) was dissolved in tetrahydrofuran (15 mL), and a 2M solution of sodium bis(trimethylsilyl)amide in tetrahydrofuran was added under ice bath. The temperature was maintained and stirred for 30 minutes. Then, a solution of crude compound 1h (2.3 g, 4.9 mmol) in tetrahydrofuran (20 mL) was added under ice bath. The temperature was maintained and stirred for 1 hour. Saturated ammonium chloride solution was added to the reaction solution for quenching, and the solution was extracted with ethyl acetate (30 mL×2). The organic phases were combined and dried over anhydrous sodium sulfate. The desiccant was filtered out and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent system B to obtain 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-oxapropane

[0166] -3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-4-carboxylic acid tert-butyl ester 1k

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

[0168] Step 8

[0169] (5S,5aS,6S,9R)-2-Chloro-1-fluoro-5-methyl-12-((1-((methylsulfonyl)oxy)methyl)cyclopropyl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-14carboxylic 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), and methanesulfonyl chloride (25 mg, 218.2 μmol) was added under ice bath. The reaction was naturally restored to room temperature for 30 minutes. Saturated aqueous ammonium chloride was added to the reaction solution for quenching, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed with water and saturated sodium chloride solution in turn, and dried over anhydrous sodium sulfate. The desiccant was filtered out and the filtrate was concentrated under reduced pressure to obtain a crude solid compound 1l (95 mg), which was used directly in the next step without purification.

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

[0173] Step 9

[0174] tert-Butyl (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-pentaaza-6,9-methanaphtho[1,8-ab]heptyl-14-carboxylate

[0175] Ester 1m

[0176] The 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]thiophen-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-pentaaza-6,9-methano[1,8-ab]heptyl-14-carboxylic acid tert-butyl ester 1o

[0179] Compound 1m (20 mg, 31.6 μmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborohexan-2-yl)-7-fluorobenzo[b]thiophen-2-yl)carbamic acid tert-butyl ester 1n (19 mg, 47 μmol, prepared by the method disclosed in preparation 23 on page 39 of the specification of patent application "WO2022261154A1"), tetrakis(triphenylphosphine)palladium (7 mg, 6.1 μmol), and cesium carbonate (30 mg, 92 μmol) were mixed in N,N-dimethylformamide (1 mL), replaced with nitrogen, and reacted at 90°C for 2 hours. After the reaction solution was cooled to room temperature, ethyl acetate was added to dilute it, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure to obtain a crude solid compound 1o (15 mg), which was directly used in the next step 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-pentaaza-6,9-methano[1,8-ab]heptyl-2-yl)benzo

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

[0184] The crude compound 1o (15 mg, 16.9 μmol) was dissolved in acetonitrile (1 mL), and 4 M hydrochloric acid 1,4-dioxane solution (0.5 mL) was added under ice bath, and the temperature was maintained for 0.5 hours, and then the reaction was carried out at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (Waters-2545, chromatographic column: ODS-BIO C18, 30*150 mm, 5 μm; mobile phase: aqueous phase (10 mmol / L ammonium bicarbonate) and acetonitrile, gradient ratio: acetonitrile 54%-69% 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 with complete medium, i.e., 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 in a 96-well low-adhesion plate (Corning, CLS7007-24EA) at a density of 1000 cells / well using complete medium, with 90 μL of cell suspension per well, centrifuged at 2000 rpm at room temperature for 5 minutes, and then placed in a 37°C, 5% CO2 cell culture incubator for overnight culture.

[0190] AGS cells were cultured with complete medium, i.e., RPMI1640 medium (Hyclone, SH30809.01) containing 10% fetal bovine serum (Corning, 35-076-CV). On the first day of the experiment, AGS cells were seeded in a 96-well low-adhesion plate (Corning, CLS7007-24EA) at a density of 1000 cells / well using complete medium, with 90 μL of cell suspension per well, centrifuged at 2000 rpm for 5 minutes at room temperature, and then placed in a 37°C, 5% CO2 cell culture incubator for overnight culture.

[0191] On the second day, 10 μL of the compound to be tested prepared with complete medium was added to each well. The final concentration of the compound for GP2d cells was 9 concentration points of 5-fold gradient dilution starting from 1 μM, and the final concentration of the compound for AGS cells was 9 concentration points of 5-fold gradient dilution starting from 10 μM. A blank control containing 0.5% DMSO was set. The well plate was placed in a cell culture incubator at 37°C and 5% CO2 for 5 days. On the seventh day, the 96-well cell culture plate was taken out and 50 μL of the compound to be tested was added to each well. 3D Cell Viability Assay reagent (Promega, G9682) was shaken at room temperature in the dark for 25 minutes, then pipetted to mix and 100 μL was taken out from each well and transferred to a white opaque 96-well plate (PerkinElmer, 6005290), and the luminescent signal value was read using a multi-function microplate reader (PerkinElmer, EnVision2105).

[0192] The IC of the inhibitory activity of the compounds was calculated using Graphpad Prism software 50 The results are shown in Table 1 below.

[0193] Table 1 AGS, GP2d cell 3D proliferation inhibitory activity data

[0194]

[0195]

[0196] Conclusion: The disclosed compounds have a good inhibitory effect on the 3D proliferation of AGS and GP2d cells.

[0197] Test Example 3: Biological evaluation of AsPC-1 cell 3D proliferation inhibition experiment

[0198] On the first day of the experiment, AsPC-1 cells that grew 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 required level. 90 μL of cell suspension was added to each well of a U-shaped low-adsorption 96-well plate (Corning, CLS7007-24EA), and the cell density was 1500 cells / well. After the cell plate was centrifuged at 2500 rpm for 5 minutes, it was placed in a 37°C, 5% CO2 incubator and cultured overnight. On the second day, the 20mM test compound dissolved in DMSO was diluted with DMSO to a first concentration of 2mM, and then diluted 5-fold gradients for a total of 9 concentration points, and the control well was DMSO. The gradient diluted compound was then further diluted 20 times with culture medium. 10 μL of the test compound diluted with culture medium was added to each well of the cell plate, and the final concentration of the compound was the first concentration of 10 μM and the 9 concentration points of the 5-fold gradient dilution. The cell wells containing 0.5% DMSO were set as the solvent control wells, and the wells containing only the culture medium and 0.5% DMSO were set as blank control wells. The compound concentrations and control wells were set in duplicate, and the final DMSO concentration in each well was 0.5%. After centrifugation at 2500 rpm for 3 minutes, the cell plate was placed in a 37°C, 5% CO2 incubator for 5 days. On the seventh day, the 96-well cell culture plate was removed and 50 μL of luminescent cell activity detection reagent was added to each well. 3D Cell Viability Assay (Promega, G9683), shake at room temperature in the dark for 25 minutes, pipette up and down to mix, and transfer 100 μL per well to the white opaque OptiPlate TM -96-well plate (PerkinElmer, 6005290), and the luminescent signal value was read using a multi-function microplate reader (PerkinElmer, EnVision2105).

[0199] The inhibition rate was calculated using the following formula: Inhibition rate = (luminescence 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%. GraphPadPrism software was used to draw a curve based on the concentration of each compound and the corresponding inhibition rate, and the IC of the compound was calculated. 50 value.

[0200] Table 2. AsPC-1 cell 3D proliferation inhibition activity data

[0201] Example No. <![CDATA[AsPC-1 / IC 50 (nM)]]> 1 2.4

[0202] Conclusion: The disclosed compounds have a good inhibitory effect on the 3D proliferation of AsPC-1 cells.

[0203] Test Example 4: Pharmacokinetic Evaluation

[0204] 1. Summary

[0205] Balb / c nude mice were used as test animals, and the drug concentration in plasma at different times after gavage (ig) of the example compounds to balb / c nude mice was determined by LC / MS / MS method. The pharmacokinetic behavior of the disclosed compounds in balb / c nude mice was studied, and their pharmacokinetic characteristics were evaluated.

[0206] 2. Experimental plan

[0207] 2.1. Experimental Drugs

[0208] Compound 1.

[0209] 2.2 Experimental Animals

[0210] 9 female BALB / c nude mice were provided by Weitonglihua Laboratory Animal Technology Co., Ltd., with production license SCXK(Zhejiang)2019-0001.

[0211] 2.3 Drug preparation

[0212] A certain amount of the test compound was weighed respectively, and 5% DMF+45% PG+50% (10% HS15-pH7.4 buffer)+400 mpk SNAC was added to prepare a 4 mg / mL colorless clear solution.

[0213] 2.4 Drug administration

[0214] The dosage was 40.0 mg / kg and the administration volume was 10 mL / kg.

[0215] 3. Operation

[0216] Before administration and 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, and 24.0 hours after administration, 0.1 mL of blood was collected from the eye sockets and placed in EDTA-K2 anticoagulant tubes. The blood was centrifuged at 10,000 rpm for 1 minute (4°C), and the plasma was separated within 1 hour and stored at -20°C for testing. The blood collection and centrifugation process was carried out under ice bath conditions.

[0217] Determine the content of the test compound in the plasma of balb / c nude mice after administration of different concentrations of the drug: Take 25 μL of balb / c nude mouse plasma samples at each time after administration, 200 μL of acetonitrile containing internal standard (Compound 1: internal standard is verapamil 100 ng / mL), vortex mix for 5 minutes, and centrifuge at 3700 rpm for 20 minutes, and mix the supernatant with water (1:1). Take 0.5-2 μL of the supernatant for LC / MS / MS analysis.

[0218] 4. Pharmacokinetic parameter results

[0219] Table 3. Pharmacokinetic parameters of the disclosed compounds in balb / c nude mice

[0220]

[0221] Conclusion: The disclosed compounds have good pharmacokinetic advantages in balb / c nude mice.

[0222] Example 2 Preparation of hydrochloride

[0223] 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 vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous hydrochloride salt, and the XRPD spectrum had no obvious characteristic peaks. The ion results showed that the chloride ion content was 8.29%.

[0224] Example 3 Preparation of hydrochloride

[0225] 7 mg of the compound of formula 1 was dissolved in 0.1 mL of the solvent in Table 4, 6 μL of 2M hydrochloric acid ethanol solution was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was amorphous hydrochloride as determined by X-ray powder diffraction.

[0226] Table 4

[0227] Solvents Crystal form acetone Hydrochloride amorphous 2-Methyltetrahydrofuran Hydrochloride amorphous

[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, 6 μL of 2M sulfuric acid ethanol solution was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction detection showed that the product was amorphous sulfate, and the XRPD spectrum had no obvious characteristic peaks. The ion results showed that the sulfate content was 12.62%.

[0230] Example 5 Preparation of sulfate

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

[0232] Table 5

[0233] Solvents 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 of Formula 1 was added with 0.15 mL of 10% water / methanol and 10.7 μL of 2M sulfuric acid ethanol solution, stirred to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as sulfate crystal form I by X-ray powder diffraction detection, and the XRPD spectrum was as follows: 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, 6 μL of 2M phosphoric acid ethanol solution was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction detection showed that the product was amorphous phosphate, and the XRPD spectrum had no obvious characteristic peaks. The ion results showed that the phosphate content was 15.03%.

[0240] Example 8 Preparation of Phosphate

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

[0242] Table 7

[0243]

[0244]

[0245] Example 9 Preparation of Phosphate Crystalline Form I

[0246] 7 mg of the amorphous compound of Formula 1 was added with 0.15 mL of 10% water / methanol and 10.7 μL of 2M phosphoric acid ethanol solution, stirred to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was phosphate crystal form I as detected by X-ray powder diffraction. The XRPD spectrum is shown in Figure 2 The positions of its characteristic peaks are shown in Table 8. The DSC spectrum shows that the endothermic peak value is 196.2°C. The TGA spectrum shows that the weight loss is 2.02% at 33°C-143°C. The ion results show that the phosphate content is 16.85%.

[0247] Table 8

[0248]

[0249] Example 10 Preparation of Methanesulfonate

[0250] 7 mg of the compound shown in formula 1 was dissolved in 0.2 mL of ethyl acetate, 6 μL of 2M methanesulfonic acid ethanol solution was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction detection showed that the product was amorphous methanesulfonate, and the XRPD spectrum had no obvious characteristic peaks. The ion results showed that the methanesulfonic acid content was 20.96%.

[0251] Example 11 Preparation of Methanesulfonate

[0252] 7 mg of the compound of formula 1 was dissolved in 0.1 mL of the solvent in Table 9, 6 μL of 2M methanesulfonic acid ethanol solution was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was amorphous methanesulfonate.

[0253] Table 9

[0254] Solvents Crystal form acetone Methanesulfonate amorphous 2-Methyltetrahydrofuran Methanesulfonate amorphous

[0255] Example 12 Preparation of Acetate

[0256] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of the solvent in Table 10, 6 μL of 2M acetic acid ethanol solution and 0.1 mL of n-heptane were added, stirred at room temperature for precipitation, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous acetate salt, and the XRPD spectrum had no obvious characteristic peaks.

[0257] Table 10

[0258] Solvents Crystal form acetone Acetate amorphous 2-Methyltetrahydrofuran Acetate amorphous

[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, 6 μL of 2M citric acid ethanol solution was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous citrate salt, and the XRPD spectrum had no obvious characteristic peaks. The 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 of formula 1 was dissolved in 0.1 mL of the solvent in Table 11, 6 μL of 2M ethanol solution of citric acid was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was amorphous citrate.

[0263] Table 11

[0264] Solvents Crystal form acetone Citrate amorphous 2-Methyltetrahydrofuran Citrate amorphous

[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, 1.4 mg of succinic acid was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was amorphous succinate, and the XRPD spectrum had no obvious characteristic peaks. The ion results showed that the succinic acid content was 17.52%.

[0267] Example 16 Preparation of succinate

[0268] 7 mg of the compound of formula 1 was dissolved in 0.1 mL of the solvent in Table 12, 1.4 mg of succinic acid was added, and the mixture was stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was an amorphous succinate salt as determined by X-ray powder diffraction.

[0269] Table 12

[0270] Solvents 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 2M p-toluenesulfonic acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was amorphous p-toluenesulfonate, and the XRPD spectrum had no obvious characteristic peaks. The ion results showed that the p-toluenesulfonic acid content was 30.49%.

[0273] Example 18 Preparation of p-toluenesulfonate

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

[0275] Table 13

[0276] Solvents 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, 6 μL of 2M maleic acid ethanol solution was added, stirred at room temperature for precipitation, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was amorphous maleate, and the XRPD spectrum had no obvious characteristic peaks. The ion results showed that the maleic acid content was 16.94%.

[0279] Example 20 Preparation of maleate

[0280] 7 mg of the compound represented by Formula 1 was dissolved in 0.1 mL of the solvent in Table 14, 6 μL of 2M maleic acid ethanol solution was added, and the mixture was stirred at room temperature to precipitate. The mixture was centrifuged and the solid was vacuum dried to obtain a solid product.

[0281] Table 14

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

[0283] Example 21 Preparation of Fumarate Crystal Form I

[0284] 7 mg of the compound of formula 1 was dissolved in 0.2 mL of ethyl acetate, 1.3 mg of fumaric acid was added, and the mixture was stirred at room temperature to precipitate. The mixture was centrifuged and the solid was dried in vacuum to obtain a solid product.

[0285] The product was defined as fumarate crystal form I by X-ray powder diffraction detection, and the XRPD spectrum was as follows: Figure 3 The characteristic peak positions are shown in Table 15. The ion results showed that the fumaric acid content was 14.23%.

[0286] Table 15

[0287]

[0288] Example 22 Preparation of Fumarate Crystal Form I

[0289] 7 mg of the compound of formula 1 was dissolved in 0.1 mL of the solvent in Table 16, 1.3 mg of fumaric acid was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was fumarate crystal form I as determined by X-ray powder diffraction.

[0290] Table 16

[0291] Solvents Crystal form acetone Fumarate Form I 2-Methyltetrahydrofuran Fumarate Form I

[0292] Example 23 Preparation of L-malate

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

[0294] Table 17

[0295] Solvents Crystal form 0.2 mL ethyl acetate L-Malate Amorphous 0.1 mL acetone L-Malate Amorphous 0.1mL 2-Methyltetrahydrofuran L-Malate Amorphous

[0296] Example 24L-Malate Form I Preparation

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

[0298] The TGA spectrum shows that the weight loss is 4.74% at 33℃-147℃. The ion results show that the L-malic acid content is 20.65%. Table 18

[0299]

[0300] Example 25L-Malate Form I Preparation

[0301] 7 mg of the compound of formula 1 was added with 0.1 mL of acetone and 10.7 μL of 2M L-malic acid ethanol solution, stirred to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product.

[0302] Example 26L-Tartrate Form I Preparation

[0303] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, and 6 μL of 2M L-tartaric acid ethanol solution was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as L-tartrate salt form I by X-ray powder diffraction detection. The XRPD spectrum is shown in Figure 5 The positions of its characteristic peaks are shown in Table 19. The DSC spectrum shows that the endothermic peaks are 102.17°C and 117.01°C. The TGA spectrum shows that the weight loss is 5.88% at 32°C-175°C. The ion results show that the L-tartaric acid content is 16.8%.

[0304] Table 19

[0305]

[0306] Example 27L-Tartrate Form I Preparation

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

[0308] Table 20

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

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

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

[0312] Example 29L-Tartrate Form II Preparation

[0313] 7 mg of the amorphous compound of Formula 1 was added with 0.15 mL of acetone and 10.67 μL of 2M L-tartaric acid ethanol solution, stirred to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as L-tartrate salt form II by X-ray powder diffraction detection, and the XRPD spectrum was as follows: Figure 6 The positions of its characteristic peaks are shown in Table 21. The DSC spectrum shows that the endothermic peaks are 105.17°C and 158.01°C. The TGA spectrum shows that the weight loss is 9.11% at 35°C-189°C. The ion results show that the L-tartaric acid content is 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, 1.5 mg of glutaric acid was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as glutaric acid salt form I by X-ray powder diffraction detection, and the XRPD spectrum was as follows: Figure 7 The positions of its characteristic peaks are shown in Table 22. The DSC spectrum shows that the endothermic peak value is 99.16°C. The TGA spectrum shows that the weight loss is 3.89% from 32°C to 164°C. The ion results show that the glutaric acid content is 16.78%.

[0318] Table 22

[0319]

[0320] Example 31 Preparation of Glutarate Form I

[0321] 7 mg of the compound of formula 1 was dissolved in 0.1 mL of the solvent in Table 23, 1.5 mg of glutaric acid was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as glutaric acid salt form I by X-ray powder diffraction detection.

[0322] Table 23

[0323] Solvents Crystal form acetone Glutarate Form I 2-Methyltetrahydrofuran Glutarate Form I

[0324] Example 32 Preparation of Glutarate Crystalline Form II

[0325] 7 mg of the amorphous compound of Formula 1 was added with 0.1 mL of ethyl acetate and 2.8 mg of glutaric acid, stirred to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as glutaric acid salt form II by X-ray powder diffraction detection, and the XRPD spectrum was as follows: Figure 8 The positions of its characteristic peaks are shown in Table 24. The DSC spectrum shows that the peak value of the endothermic peak is 111.16°C. The TGA spectrum shows that the weight loss is 5.63% at 32°C-139°C. The ion results show that the glutaric acid content is 26%.

[0326] Table 24

[0327]

[0328] Example 33 Preparation of Benzoate Crystal Form I

[0329] 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 the solid was vacuum dried to obtain a solid product. The product was defined as benzoate crystal form I by X-ray powder diffraction detection. The XRPD spectrum is shown as follows: Fig. 9 The positions of its characteristic peaks are shown in Table 25. The DSC spectrum shows that the endothermic peak value is 169°C. The TGA spectrum shows that the weight loss is 1.95% at 32°C-150°C. The ion results show that the benzoic acid content is 14.93%.

[0330] Table 25

[0331]

[0332] Example 34 Preparation of Benzoate Crystal Form I

[0333] 7 mg of the compound of formula 1 was dissolved in 0.1 mL of the solvent in Table 26, 6 μL of 2M benzoic acid ethanol solution was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was benzoate crystal form I as determined by X-ray powder diffraction.

[0334] Table 26

[0335] Solvents Crystal form acetone Benzoate salt form I 2-Methyltetrahydrofuran Benzoate salt form I

[0336] Example 35 Preparation of o-benzoylsulfonyl imide complex

[0337] 7 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of ethyl acetate, 2.1 mg of o-benzoylsulfonyl imide was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as an amorphous o-benzoylsulfonyl imide complex by X-ray powder diffraction detection, and the XRPD spectrum had no obvious characteristic peaks.

[0338] Example 36 Preparation of o-benzoylsulfonyl imide complex

[0339] 7 mg of the compound of formula 1 was dissolved in 0.1 mL of 2-methyltetrahydrofuran, 2.1 mg of o-benzoylsulfonyl imide was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. X-ray powder diffraction analysis showed that the product was an amorphous o-benzoylsulfonyl imide complex.

[0340] Example 37 Preparation of o-benzoylsulfonyl imide complex crystal form I

[0341] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of acetone, and 2.1 mg of o-benzoylsulfonyl imide was added. The mixture was stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as o-benzoylsulfonyl imide complex crystal form I by X-ray powder diffraction detection. The XRPD spectrum is shown as follows: Fig.10 The positions of its characteristic peaks are shown in Table 27. The DSC spectrum shows that the endothermic peaks are 68.18°C, 106.18°C and 160.02°C. The TGA spectrum shows that the weight loss is 3.92% at 31°C-115°C. The NMR results show that the ratio of the compound to o-benzoylsulfonyl imide is 1:1.

[0342] Table 27

[0343]

[0344]

[0345] Example 38 Preparation of o-benzoylsulfonyl imide complex crystal form I

[0346] 7 mg of the amorphous compound of Formula 1 was added with 0.15 mL of 10% water / methanol and 4.2 mg of o-benzoylsulfonyl imide, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product.

[0347] Example 39 Preparation of Arginine Complex Crystalline Form a

[0348] 7 mg of the compound shown in Formula 1 was dissolved in 0.1 mL of acetone, 1.9 mg of arginine was added, stirred at room temperature to precipitate, centrifuged, and the solid was vacuum dried to obtain a solid product. The product was defined as arginine complex crystal form a by X-ray powder diffraction detection, and the XRPD spectrum was as follows: Fig.11 The positions of its characteristic peaks are shown in Table 28. The DSC spectrum shows that the endothermic peaks are 68.18°C, 106.18°C and 160.02°C. The TGA spectrum shows that the weight loss is 3.92% at 31°C-115°C.

[0349] Table 28

[0350]

[0351] Example 40 Influencing Factors Stability

[0352] The above-mentioned o-benzoylsulfonyl imide complex crystal form I was laid open and placed flat, and the stability of the sample was investigated under light (4500 Lux), high temperature (40° C., 60° C.), and high humidity (RH 75%, RH 92.5%) conditions, and the sampling period was 30 days.

[0353] Table 28

[0354]

[0355]

[0356] Conclusion: The influencing factor experiment shows that under high temperature 40°C and 60°C and high humidity 75% and 92.5% for 30 days, the physical and chemical stability of the disclosed crystal form is good.

[0357] Example 41: Long-term accelerated stability test

[0358] The above-mentioned o-benzoylsulfonyl imide complex crystal form I was placed at 25°C / 60%RH and 40°C / 75%RH to investigate the stability.

[0359] Table 29

[0360]

[0361] Conclusion: Long-term accelerated experiments show that the physicochemical stability of the disclosed crystal form is good under the conditions of 25°C / 60% RH and 40°C / 75% RH for 1 month.

Claims

1. A complex 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-methylnaphtho[1,8-ab]heptylcyclo-2-yl)benzo[b]thiophene-3-carbonitrile as shown in formula 1, wherein the ligand of the complex is selected from arginine, o-benzoylsulfonimide, 2. The o-benzoylsulfonyl imide complex crystalline form I of the compound of formula 1 according to claim 1, characterized in that: The 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, preferably at 6.985, 8.161, 11.381, 12.871, 14.733, 15.137, 16.472, 18.232, 19.186, 23.303, and 25.891 , and 29.367, and more preferably have 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, and 29.

367.

3. The o-benzoylsulfonyl imide complex crystalline form I according to claim 2, characterized in that: The X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ is shown in FIG10 .

4. A method for preparing the o-benzoylsulfonyl imide complex crystalline form I of the compound of formula 1 as claimed in claim 2 or 3, the method comprising the steps of dissolving the compound of formula 1 in acetone, adding o-benzoylsulfonyl imide, and stirring.

5. The arginine complex crystal form a of the compound of formula 1 according to claim 1, characterized in that: The X-ray powder diffraction pattern expressed by the diffraction angle 2θ has characteristic peaks at 15.849, 18.709, 19.847, 20.783, 23.275, 26.583, and 32.

305.

6. The arginine complex crystal form a according to claim 5, characterized in that: The X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ is shown in FIG11 .

7. A method for preparing the arginine complex crystal form a according to any one of claims 5 to 6, comprising the steps of dissolving the compound of formula 1 in acetone, adding arginine, and stirring.

8. According to the crystal form according to any one of claims 2-3, 5-6, the 2θ angle error range is ±0.

20.

9. A pharmaceutical composition comprising the complex of the compound of formula 1 as claimed in claim 1, or the crystalline 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: A step of mixing the complex of the compound of formula 1 according to claim 1, or the crystalline form according to any one of claims 2-3, 5-6 and a pharmaceutically acceptable excipient.

11. Use of the complex of the compound of formula 1 according to claim 1, or the crystalline 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 the complex 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 medicament for treating and / or preventing cancer.

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