Salt form of pyridazine derivative, crystal form, method for preparing same, and use thereof

Abstract

The present invention relates to the field of pharmaceuticals, and in particular, to a salt form of a pyridazine derivative, a crystal form, a method for preparing same, and use thereof. The present invention provides a salt form of a compound of formula I, and particularly relates to a hydrochloride salt, a hydrobromide salt, a sulfate salt, a phosphate salt, an oxalate salt, a citrate salt, a malate salt, a maleate salt, a fumarate salt, a succinate salt, a malonate salt, a methanesulfonate salt, a benzenesulfonate salt, and a p-toluenesulfonate salt. The present invention further provides an amorphous form and a crystal form of the hydrochloride salt, and particularly relates to crystal form A and crystal form B. The present invention further provides a method for preparing the salt form and the crystal form, and use thereof. The salt form of the compound of formula I and the crystal form of the present invention have significant SOS1 inhibitory activity and a significant anti-tumor cell proliferation effect, have advantageous pharmaceutical properties such as high solubility, good stability, and excellent pharmacokinetics, and are thus suitable for pharmaceutical use.

Description

Salt forms, crystal forms, preparation methods and applications of pyridazine derivatives Technical Field

[0001] This invention relates to the salt forms, crystal forms, preparation methods, and applications of pyridazine derivatives, and belongs to the pharmaceutical field. Background Technology

[0002] Rat sarcoma (RAS) is the most frequently mutated oncogene in human cancers, accounting for approximately 30% of all cancer mutations. RAS acts as a molecular switch, switching between an inactive state (binding to GDP) and an activated state (binding to GTP). This process is regulated by GTPase activating proteins (GAPs) and guanine nucleotide exchange factors (GEFs). SOS (son of sevenless) is a crucial guanine nucleotide exchange factor (GEF) for activating RAS proteins in cells. It consists of two parts, SOS1 and SOS2. Although their structures are highly homologous, current research indicates that SOS1 plays a more significant role than SOS2 in controlling cell proliferation and migration. Therefore, current research focuses primarily on targeting SOS1. The SOS1 protein is 152 kDa in size and contains 1333 amino acid residues. The SOS1 protein domain is divided into three parts: the N-terminal region, the catalytic and allosteric domain, and the C-terminal region. These regions work together to regulate the GEF activity of SOS1. When receptor tyrosine kinases (RTKs), primarily epidermal growth factor receptor kinase (EGFR), are activated on the cell membrane, they dissociate phosphate groups and transfer them to growth factor receptor-binding protein 2 (GRB2), activating it. Activated GRB2 recruits SOS1 to the plasma membrane, activating SOS1. Activated SOS1 binds to RAS-GDP, forming RAS-GTP through nucleotide exchange, thereby activating two key downstream signaling pathways: RAS-RAF-MAPK and RAS-PI3K-AKT-mTOR. This is a crucial hub for the regulation of RTK-RAS and downstream signaling pathways, playing a vital role in tumor development, progression, migration, and prognosis. Besides tumors, studies have shown that some pathogenic skin diseases, such as Noonan syndrome (NS), cardiofacial syndrome (CFC), and hereditary gingival fibromatosis type I, are also closely related to SOS1.

[0003] In recent years, research on KRAS proteins and drugs has attracted great interest. Currently, research on KRAS... G12CSeveral drugs have been launched and achieved positive therapeutic effects for the KRAS mutation type, but there are still no effective drugs for tumors with other KRAS protein mutation types. From the perspective of SOS1's mechanism of action, SOS1 inhibitors can target multiple KRAS mutant subtypes (including KRAS...). G12C KRAS G12D KRAS G12V These drugs exhibit inhibitory activity and belong to the pan-KRAS inhibitor class. Developing novel SOS1 inhibitors holds promise for providing new treatment options for various KRAS-mutated tumors. Currently, the development of SOS1 inhibitors has made some progress in the pharmaceutical industry; for example, Boehringer Ingelheim's BI-1701963 has entered Phase I clinical trials, and Bristol-Myers Squibb's MRTX0902 is in Phase I / II clinical trials. However, these drugs all have issues with safety and poor admixture properties, and there is still an urgent need to develop novel SOS1 inhibitors with high activity, high selectivity, good safety, and favorable pharmacokinetic properties. Summary of the Invention

[0004] This invention aims to solve at least one of the aforementioned technical problems, thereby improving the pharmaceutical properties of the compound. To this end, this invention has the following four objectives. The first objective is to provide a salt form of a pyridazine derivative having SOS1 inhibitory activity. The second objective is to provide a crystalline form (including an amorphous form) of the hydrochloride salt of said pyridazine derivative. The third objective is to provide a method for preparing said salt form and crystalline form. The fourth objective is to provide applications of said salt form and crystalline form.

[0005] This invention provides hydrochloride salts of the compounds shown in Formula I:

[0006] Preferably, the molar ratio of the compound of formula I to hydrochloric acid in the hydrochloride is 1:2.

[0007] The present invention provides hydrobromide salts of the compounds shown in Formula I.

[0008] Preferably, the molar ratio of the compound of formula I to hydrobromic acid in the hydrobromate is 1:1.7.

[0009] The present invention provides sulfates of the compounds shown in Formula I.

[0010] Preferably, the molar ratio of the compound of formula I to sulfuric acid in the sulfate is 1:1.2.

[0011] This invention provides phosphates of the compounds shown in Formula I.

[0012] Preferably, the molar ratio of the compound represented by Formula I to phosphoric acid in the phosphate is 1:2.

[0013] This invention provides oxalate salts of the compounds shown in Formula I.

[0014] Preferably, the molar ratio of the compound of formula I to oxalic acid in the oxalate is 1:1.8.

[0015] The present invention provides citrate salts of the compounds shown in Formula I.

[0016] Preferably, the molar ratio of the compound of formula I to citric acid in the citrate is 1:1.8.

[0017] This invention provides malates of the compounds shown in Formula I.

[0018] Preferably, the molar ratio of the compound of formula I to malic acid in the malate is 1:1.8.

[0019] The present invention provides maleate salts of the compounds shown in Formula I.

[0020] Preferably, the molar ratio of the compound of formula I to maleic acid in the maleate salt is 1:2.

[0021] This invention provides fumarate salts of the compounds shown in Formula I.

[0022] Preferably, the molar ratio of the compound represented by Formula I to fumaric acid in the fumarate is 1:1.3.

[0023] The present invention provides succinate of the compound shown in Formula I.

[0024] Preferably, the molar ratio of the compound of formula I to succinic acid in the succinate is 1:1.5.

[0025] The present invention provides a malonate of the compound shown in Formula I.

[0026] Preferably, the molar ratio of the compound of formula I to malonic acid in the malonate is 1:1.6.

[0027] The present invention provides a methanesulfonate of the compound shown in Formula I.

[0028] Preferably, the molar ratio of the compound of formula I to methanesulfonic acid in the methanesulfonate is 1:2.

[0029] The present invention provides benzenesulfonate salts of the compounds shown in Formula I.

[0030] Preferably, the molar ratio of the compound of formula I to benzenesulfonic acid in the benzenesulfonate is 1:2.

[0031] The present invention provides p-toluenesulfonate salts of the compounds shown in Formula I.

[0032] Preferably, the molar ratio of the compound of formula I to p-toluenesulfonate is 1:2.

[0033] The present invention provides an amorphous form of the hydrochloride salt.

[0034] Furthermore, the X-ray powder diffraction pattern of the amorphous form is shown in Figure 3.

[0035] The present invention provides the crystalline form of the hydrochloride.

[0036] Furthermore, the crystal form is selected from anhydrous crystal form, hydrate crystal form, and solvate crystal form.

[0037] Furthermore, the crystalline form is crystal form A, whose X-ray powder diffraction pattern has the strongest characteristic diffraction peak at the following 2θ angle: 24.24±0.2°.

[0038] Furthermore, the crystalline form is crystal form A, whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.68±0.2°, 11.01±0.2°, 14.64±0.2°, 18.70±0.2° and 24.24±0.2°.

[0039] Furthermore, the X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 11.01±0.2°, 12.70±0.2°, 14.64±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, and 24.24±0.2°.

[0040] Furthermore, the X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 11.01±0.2°, 12.70±0.2°, 14.64±0.2°, 18.25±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, 24.24±0.2°, and 24.65±0.2°.

[0041] Furthermore, the X-ray powder diffraction pattern of crystal form A also exhibits characteristic diffraction peaks at the following 1 to 10 2θ angles: 10.40±0.2°, 12.26±0.2°, 13.24±0.2°, 13.75±0.2°, 15.21±0.2°, 16.44±0.2°, 22.16±0.2°, 25.45±0.2°, 27.85±0.2°, and 29.05±0.2°.

[0042] Furthermore, the X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 10.40±0.2°, 11.01±0.2°, 12.26±0.2°, 12.70±0.2°, 13.24±0.2°, 13.75±0.2°, 14.64±0.2°, 18.25±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, 24.24±0.2°, 24.65±0.2°, 25.45±0.2°, and 29.05±0.2°.

[0043] Furthermore, the X-ray powder diffraction pattern of crystal form A exhibits characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 10.40±0.2°, 11.01±0.2°, 12.26±0.2°, 12.70±0.2°, 13.24±0.2°, 13.75±0.2°, 14.64±0.2°, 15.21°. ±0.2°, 16.44±0.2°, 17.37±0.2°, 18.25±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, 22.16±0.2°, 23.04±0.2°, 24.24±0.2°, 24.65±0.2°, 25.45±0.2°, 27.85±0.2° and 29.05±0.2°.

[0044] Furthermore, the crystalline form is crystal form A, and its X-ray powder diffraction pattern is shown in Figure 1.

[0045] Furthermore, the crystal form is crystal form A, and its differential scanning calorimetry spectrum is shown in Figure 4.

[0046] Furthermore, the crystalline form is crystal form A, and its thermogravimetric analysis spectrum shows a weight loss of 3% when heated to 110±5℃.

[0047] Furthermore, the crystal form is crystal form A, and its thermogravimetric analysis spectrum is shown in Figure 6.

[0048] Furthermore, the crystalline form is crystal form B, and its X-ray powder diffraction pattern has the strongest characteristic diffraction peak at the following 2θ angle: 23.03±0.2°.

[0049] Furthermore, the crystalline form is crystal form B, whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 9.38±0.2°, 14.64±0.2°, 23.03±0.2° and 25.80±0.2°.

[0050] Furthermore, the X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 9.38±0.2°, 14.64±0.2°, 18.34±0.2°, 23.03±0.2° and 25.80±0.2°.

[0051] Furthermore, the X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 9.38±0.2°, 14.64±0.2°, 17.43±0.2°, 18.34±0.2°, 19.73±0.2°, 23.03±0.2°, 24.64±0.2°, 25.80±0.2°, and 29.90±0.2°.

[0052] Furthermore, the X-ray powder diffraction pattern of crystal form B also exhibits characteristic diffraction peaks at the following 1 to 11 2θ angles: 8.29±0.2°, 10.99±0.2°, 11.84±0.2°, 12.25±0.2°, 12.83±0.2°, 15.05±0.2°, 15.45±0.2°, 16.60±0.2°, 22.45±0.2°, 29.20±0.2°, and 30.50±0.2°.

[0053] Furthermore, the X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 8.29±0.2°, 9.38±0.2°, 10.99±0.2°, 12.83±0.2°, 14.64±0.2°, 16.60±0.2°, 17.43±0.2°, 18.34±0.2°, 19.73±0.2°, 20.75±0.2°, 23.03±0.2°, 24.64±0.2°, 25.80±0.2°, 29.20±0.2°, and 29.90±0.2°.

[0054] Furthermore, the X-ray powder diffraction pattern of crystal form B exhibits characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 8.29±0.2°, 9.38±0.2°, 10.99±0.2°, 11.84±0.2°, 12.83±0.2°, 14.64±0.2°, 15.05±0.2°, 15.45±0.2°, 16.60±0.2°, 17.43±0.2°, 17.75 ±0.2°, 18.34±0.2°, 18.64±0.2°, 19.55±0.2°, 19.73±0.2°, 20.75±0.2°, 21.05±0.2°, 21.33±0.2°, 23.03±0.2°, 24.64±0.2°, 25.80±0.2°, 26.45±0.2°, 28.25±0.2°, 29.20±0.2°, 29.90±0.2°.

[0055] Furthermore, the crystal form is crystal type B, and its X-ray powder diffraction pattern is shown in Figure 2.

[0056] Furthermore, the crystal form is crystal form B, and its differential scanning calorimetry spectrum is shown in Figure 5.

[0057] Furthermore, the crystalline form is crystal form B, and its thermogravimetric analysis spectrum shows a weight loss of 4% when heated to 120±5℃.

[0058] Furthermore, the crystal form is crystal form B, and its thermogravimetric analysis spectrum is shown in Figure 7.

[0059] Furthermore, the X-ray powder diffraction pattern as described in any of the preceding items is obtained under Cu Kα radiation conditions.

[0060] The present invention provides a method for preparing the crystalline form described above, wherein the hydrochloride salt of the compound shown in Formula I is recrystallized with ethanol and methyl tert-butyl ether to obtain crystal form A.

[0061] Further, the preparation method includes the following steps: dissolving the hydrochloride salt of the compound shown in Formula I in ethanol, then adding methyl tert-butyl ether, crystallizing, collecting the solid, and obtaining crystal form A.

[0062] Furthermore, the mass ratio of hydrochloride to ethanol of the compound shown in Formula I is 1:(4.7–7.9).

[0063] Preferably, the mass ratio of the hydrochloride salt to ethanol of the compound shown in Formula I is 3.8:24.

[0064] Furthermore, the mass ratio of hydrochloride to methyl tert-butyl ether of the compound shown in Formula I is 1:(5.9–11.8).

[0065] Preferably, the mass ratio of the hydrochloride salt to methyl tert-butyl ether of the compound shown in Formula I is 3.8:22.6.

[0066] Further, the hydrochloride salt of the compound shown in Formula I is dissolved by heating to 70–80 °C.

[0067] Further, methyl tert-butyl ether is added after cooling to 30–50°C.

[0068] Further, crystals are precipitated by cooling the temperature to 15–35°C.

[0069] This invention provides a method for preparing the crystalline form described above, wherein the hydrochloride crystal form A of the compound shown in Formula I is recrystallized with ethanol to obtain crystal form B.

[0070] Furthermore, the preparation method includes the following steps: dissolving the hydrochloride crystal form A of the compound shown in Formula I in ethanol, then crystallizing, collecting the solid, and obtaining crystal form B.

[0071] Furthermore, the mass-to-volume ratio of the hydrochloride crystal form A of the compound shown in Formula I to ethanol is 30:240, g / mL.

[0072] Further, the temperature is raised to reflux to dissolve the hydrochloride crystal form A of the compound shown in Formula I.

[0073] Further, crystals are precipitated by cooling the temperature to 15–35°C.

[0074] The present invention provides a pharmaceutical composition comprising the salt, the amorphous form, or the crystalline form described herein, and pharmaceutically acceptable excipients or auxiliary ingredients.

[0075] The present invention provides the use of the salt, the amorphous form, the crystalline form, or the pharmaceutical composition thereof in the preparation of SOS1 inhibitors.

[0076] The present invention provides the use of the salt, the amorphous form, the crystalline form, or the pharmaceutical composition thereof in the preparation of a medicament for treating SOS1-mediated diseases.

[0077] Furthermore, the disease is selected from at least one of cancer and pathogenic skin diseases.

[0078] Furthermore, the cancer is selected from non-small cell lung cancer, lung cancer, pancreatic cancer, ovarian cancer, bladder cancer, prostate cancer, chronic myeloid leukemia, colorectal cancer, brain cancer, liver cancer, kidney cancer, stomach cancer, and breast cancer;

[0079] The pathogenic skin rashes mentioned are selected from Noonan syndrome, cardiofacial skin syndrome, and type I hereditary gingival fibromatosis.

[0080] The present invention provides the use of the salt, the amorphous form, the crystalline form, or the pharmaceutical composition thereof in the preparation of a medicament for treating diseases caused by overexpression of the SOS1 protein.

[0081] The present invention provides the use of the salt, the amorphous form, the crystalline form, or the pharmaceutical composition thereof in the preparation of a medicament for treating diseases caused by SOS1 protein overexpression.

[0082] This invention provides different salt forms and crystal forms of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one. Experiments have demonstrated that the salt forms and crystal forms provided by this invention exhibit significant SOS1 inhibitory activity and significant anti-tumor cell proliferation effects. Furthermore, they possess favorable pharmaceutical properties such as high solubility, good stability, and excellent pharmacokinetic properties, making them suitable for pharmaceutical applications. Attached Figure Description

[0083] Figure 1 shows the XRPD spectrum of crystal form A in Example 16;

[0084] Figure 2 shows the XRPD spectrum of crystal form B in Example 17;

[0085] Figure 3 shows the XRPD spectrum of the amorphous material in Example 18;

[0086] Figure 4 shows the DSC spectrum of crystal form A in Example 16;

[0087] Figure 5 shows the DSC spectrum of crystal form B in Example 17;

[0088] Figure 6 shows the TGA spectrum of crystal form A in Example 16;

[0089] Figure 7 shows the TGA spectrum of crystal form B in Example 17. Detailed Implementation

[0090] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0091] The starting materials used in the embodiments of the present invention are known and can be purchased on the market or synthesized according to known methods in the art.

[0092] Unless otherwise specified, the solvents used in this invention are those that are commercially available.

[0093] Unless otherwise specified in the examples, the reaction temperature is room temperature, ranging from 15°C to 35°C.

[0094] The X-ray powder diffractometer (XRD) method of this invention is as follows:

[0095] Instrument Model: DX-2700

[0096] Test method: Approximately 110-150 mg of sample is used for XRD detection.

[0097] The detailed XRD parameters are as follows:

[0098] X-ray source: Cu, Kα (Kα1 = 1.54056 Å, Kα2 = 1.54439 Å, Kα2 / Kα1 intensity ratio 100:50)

[0099] Phototube voltage: 40kV

[0100] Phototube current: 30mA

[0101] Diverging slit: Fixed at 1deg

[0102] Receiving slit: 0.3mm

[0103] Anti-scattering slit: 1deg

[0104] Measurement time: 17 min

[0105] Scanning angle range: 3-53 degrees

[0106] Step width angle: 0.05deg

[0107] Step length: 1 second

[0108] Sample disk rotation speed: None rpm

[0109] The differential scanning calorimeter (DSC) and thermogravimetric analysis (TGA) methods of this invention are as follows:

[0110] Instrument Model: TGA / DSC 2 Thermogravimetric Differential Scanning Calorimeter

[0111] Test method: Take a sample (about 1-10 mg) and place it in a TGA / DSC test dish. Under N2 conditions of 50 mL / min, heat the sample from 40 °C to 350 °C at a heating rate of 10 °C / min.

[0112] In this invention, NMR measurements were performed using an AVANCE NEO 400MHz Bruker instrument. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard. The chemical shifts (δ) of the NMR are given in parts per million (ppm). MS measurements were performed using an ISQ-EC Thermo Fisher LC-MS instrument.

[0113] The method for detecting anion content in this invention is as follows:

[0114] Anion content detection instruments: Thermo Fisher ICS-6000 ion chromatograph, 0.0001 g electronic balance.

[0115] Chromatographic conditions 1 for anion content detection:

[0116] Column: Dionex Ionpac™ AS11-HC, 4 x 250 mm

[0117] Flow rate: 1 mL / min

[0118] Column temperature: 30℃

[0119] Equilibrium temperature: 30℃

[0120] Suppressor model: Anion suppressor, Dinnex ASRS 300, 4mm (ERS-4mm)

[0121] Suppressor current: 99mA

[0122] Detector: Conductivity detector

[0123] Injection volume: 25 μl

[0124] Running time: 40 minutes

[0125] Rinse solution: potassium hydroxide solution

[0126] Gradient procedure:

[0127] Solution preparation:

[0128] Solvent: Deionized water

[0129] Blank solution: Take the solvent and analyze it using chromatographic conditions 1.

[0130] Oxalate ion control solution: Take an appropriate amount of oxalic acid, accurately weigh it, dissolve it in solvent and quantitatively dilute it to prepare a solution containing about 0.2 mg per 1 mL, and analyze it using chromatographic conditions 1.

[0131] Sulfate ion control solution: Take an appropriate amount of sulfuric acid, accurately weigh it, dissolve it in solvent and quantitatively dilute it to prepare a solution containing about 0.2 mg per 1 mL, and analyze it using chromatographic conditions 1.

[0132] Phosphate ion control solution: Take an appropriate amount of phosphoric acid, accurately weigh it, dissolve it in solvent and quantitatively dilute it to prepare a solution containing about 0.2 mg per 1 mL, and analyze it using chromatographic conditions 1.

[0133] Hydrobromate ion control solution: Take an appropriate amount of hydrobromic acid, accurately weigh it, dissolve it in solvent and quantitatively dilute it to prepare a solution containing about 0.5 mg per 1 mL, and analyze it using chromatographic conditions 1.

[0134] Test solution: Accurately weigh an appropriate amount of this product, dissolve it in solvent, and quantitatively dilute it to prepare a solution containing approximately 0.06 mg per mL. Inject and analyze using chromatographic conditions 1, and calculate the concentration based on peak area using the external standard method.

[0135] Chromatographic conditions 2 for anion content detection:

[0136] Column: Dionex Ionpac™ AS11-HC, 4 x 250 mm

[0137] Flow rate: 1 mL / min

[0138] Column temperature: 30℃

[0139] Equilibrium temperature: 35℃

[0140] Suppressor model: Anion suppressor, Dinnex ASRS 300, 4mm (ERS-4mm)

[0141] Suppressor current: 33mA

[0142] Detector: Conductivity detector

[0143] Injection volume: 25 μl

[0144] Running time: 15min

[0145] Rinse solution: potassium hydroxide solution

[0146] Gradient procedure:

[0147] Solution preparation:

[0148] Solvent: Deionized water

[0149] Blank solution: Take the solvent and analyze it using chromatographic conditions 2.

[0150] Chloride ion control solution: Take an appropriate amount of chloride standard solution in water, accurately transfer it, dissolve it in solvent and quantitatively dilute it to prepare a solution containing about 10 μg per 1 mL, and analyze it using chromatographic conditions 2.

[0151] Test solution: Accurately weigh an appropriate amount of this product, dissolve it in solvent, and quantitatively dilute it to prepare a solution containing approximately 80 μg per mL. Inject and analyze using chromatographic conditions 2, and calculate the concentration based on peak area using the external standard method.

[0152] Example 1: Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one

[0153] Step a): Preparation of 3-((1-methylcyclopropyl)amino)acrylonitrile

[0154] Methanol (70 kg) and 1-methylcyclopropane-1-amine hydrochloride (30 kg) were added to a 300 L reactor at room temperature. Triethylamine (56.4 kg) was added with stirring, and the mixture was stirred until homogeneous. The temperature was raised to 55 °C, and isoxazole (20.3 kg) was added dropwise. After the addition was complete, the mixture was refluxed for 12 h. After the reaction was completed, the reaction solution was cooled to 40 °C, concentrated under reduced pressure, and the residue was dissolved in water (150 kg). Dichloromethane (200 kg) was added, and the mixture was stirred at room temperature for 10 min. The mixture was separated, and the aqueous phase was extracted with dichloromethane (120 kg). The organic phases were combined, and saturated brine (230 kg) was added to the organic phase. The mixture was stirred for 30 min, allowed to stand, and then separated. Anhydrous sodium sulfate (15 kg) was added to the organic phase, and the mixture was stirred for 2 h. The mixture was filtered under reduced pressure, and the filter cake was washed with dichloromethane (34 kg). The filtrates were combined and concentrated under reduced pressure to obtain 3-((1-methylcyclopropyl)amino)acrylonitrile. No further purification is required; it can be used directly in the next reaction.

[0155] Step b): Preparation of methyl 5-cyano-1-(1-methylcyclopropyl)-2-oxo-1,2-dihydropyridine-4-carboxylate

[0156] Dichloromethane (60 kg) and 3-((1-methylcyclopropyl)amino)acrylonitrile (3.92 kg) were added to a 100 L reactor. Dimethyl butyrynethide (11.4 kg) was added with stirring. The reactor was cooled to -10 °C, and DBU (4.9 kg) was slowly added dropwise. After the addition was complete, the reactor was allowed to naturally warm to room temperature and react for 3 hours. At the end of the reaction, 18 kg of 2.5% hydrogen chloride aqueous solution was added, and the mixture was stirred for 30 minutes. The mixture was then allowed to stand and the layers were separated. The organic phase was washed successively with saturated sodium bicarbonate aqueous solution (20 kg) and saturated brine (23 kg), dried with anhydrous sodium sulfate (4 kg), filtered under reduced pressure, and the filter cake was rinsed with dichloromethane (12 kg). The filtrates were combined and concentrated under reduced pressure. The residue was dissolved by stirring with ethyl acetate (12.5 kg) at 40 °C. Methyl tert-butyl ether (25 kg) was slowly added and stirred for 4 h. After centrifugation, the resulting solid was slurried with 10% methanol aqueous solution (40 kg) for 12 h, centrifuged again, rinsed with 10% methanol aqueous solution (8 kg), and the solid was collected and dried under vacuum for 12 h to obtain methyl 5-cyano-1-(1-methylcyclopropyl)-2-oxo-1,2-dihydropyridine-4-carboxylate. No further purification was required, and it was used directly in the next reaction.

[0157] Step c): Preparation of 3-amino-5-(1-methylcyclopropyl)-1H-pyrrolo[3,4-c]pyridine-1,6(5H)-dione

[0158] 7.45 kg of methyl 5-cyano-1-(1-methylcyclopropyl)-2-oxo-1,2-dihydropyridine-4-carboxylate and 22 kg of methanol were added to a 50 L reactor. Ammonia-methanol solution (11.54 kg) was added with stirring, and the reaction was carried out at 25 °C for 20 h. The mixture was filtered under reduced pressure, and the filter cake was washed with methanol (7.5 kg). The solid was collected and dried under vacuum to give 3-amino-5-(1-methylcyclopropyl)-1H-pyrrolo[3,4-c]pyridine-1,6(5H)-dione. The overall yield of the three steps (ac) was 66.0%.

[0159] Step d): Preparation of (R)-3-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-5-(1-methylcyclopropyl)-1H-pyrrolo[3,4-c]pyridine-1,6(5H)-dione hydrochloride

[0160] Isopropanol (18 kg) was added to a 100 L reactor. Under stirring, 3-amino-5-(1-methylcyclopropyl)-1H-pyrrolo[3,4-c]pyridine-1,6(5H)-dione (4.5 kg) and (R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethane-1-amine hydrochloride (5.2 kg) were added in sequence. After the addition was complete, the temperature was raised to 80 °C and the reaction was maintained for 20 h. After the reaction was completed, the reaction solution was cooled to 45-55°C, and n-heptane (77 kg) was added under stirring. The solution was then cooled to 10-20°C and stirred for 2 hours. After centrifugation, the solution was washed with n-heptane (6.2 kg), the solid was collected, and dried under vacuum to give (R)-3-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-5-(1-methylcyclopropyl)-1H-pyrrolo[3,4-c]pyridine-1,6(5H)-dione hydrochloride, with a yield of 89.0%.

[0161] Step e): Preparation of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-2,6-dihydropyrido[3,4-d]pyridazine-1,7-dione

[0162] Ethanol (160.5 kg) was added to a 300 L reactor. (R)-3-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-5-(1-methylcyclopropyl)-1H-pyrrolo[3,4-c]pyridine-1,6(5H)-dione hydrochloride (4 kg) and DBU (2.9 kg) were added while stirring. After the mixture was dissolved, hydrazine hydrate (0.87 kg) was added. The mixture was stirred and reacted at 25-30 °C for 60 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was diluted with ethanol (3.2 kg). The diluted solution was added to purified water (120 kg) under stirring at 20-30 °C. After the addition was complete, the mixture was centrifuged and washed with purified water (12 L). The solid was collected and dried under vacuum for 24 h to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-2,6-dihydropyrido[3,4-d]pyridazine-1,7-dione, with a yield of 64.2%.

[0163] Step f): Preparation of (R)-1-chloro-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)pyrido[3,4-d]pyridazine-7(6H)-one

[0164] Add 23 kg of 1,4-dioxane to a 100 L reactor, then add (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-2,6-dihydropyrido[3,4-d]pyridazine-1,7-dione (4.5 kg) while stirring, followed by 2.7 kg of phosphorus oxychloride. Heat to 100 °C and stir for 24 h. After the reaction is complete, cool the reaction solution to room temperature, slowly add 60 kg of dichloromethane, then slowly add 10% sodium bicarbonate aqueous solution to quench the reaction, add 8.8 kg of isopropanol and stir to extract, allow to stand and separate, and collect the organic phase. Extracted again with a suitable amount of dichloromethane / isopropanol mixed solvent (v / v = 5:1), the organic phases were combined, washed with saturated salt water, dried with anhydrous sodium sulfate (9 kg), filtered under reduced pressure, the filter cake was washed with dichloromethane (10 kg), the filtrates were combined, concentrated under reduced pressure, the residue was added with ethyl acetate (8.1 kg) and stirred at 40-50 °C to precipitate crystals. After a large amount of solid precipitated, n-heptane (12.3 kg) was added at 30-50 °C. After the addition was complete, the mixture was cooled to 10-20 °C and stirred for 2 h. The mixture was filtered, the filter cake was washed with n-heptane (6 kg), the filter cake was collected, and dried under vacuum to give (R)-1-chloro-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)pyrido[3,4-d]pyridazine-7(6H)-one, with a yield of 92.1%.

[0165] Step g): Preparation of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one

[0166] Isopropanol (25.7 kg) was added to a 50 L reactor. While stirring, (R)-1-chloro-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)pyrido[3,4-d]pyridazine-7(6H)-one (4.11 kg) and N-methylpiperazine (1.2 kg) were added. The mixture was heated to reflux for 48 h. After the reaction was complete, the reaction solution was cooled to 10–40 °C, and 4.1 L of ethyl hydrogen chloride solution was added. After the addition was complete, the mixture was cooled to 10–30 °C. The resulting solution was added to methyl tert-butyl ether (120.5 kg) at 10–30 °C with stirring. The mixture was centrifuged under nitrogen protection, washed with methyl tert-butyl ether (6.2 kg), and the solid was collected. The obtained solid was dissolved in a mixed solvent of dichloromethane / ethanol (v / v = 4 / 1) (31 kg) by stirring. A 10% sodium bicarbonate aqueous solution was slowly added and stirred for 5 min. The mixture was allowed to stand and separated, and the organic phase was collected. The aqueous phase was extracted with a dichloromethane / ethanol (v / v = 4 / 1) mixed solvent (31 kg), the organic phases were combined, washed with saturated brine (44 kg), dried over anhydrous sodium sulfate, filtered under reduced pressure, the filter cake was rinsed with dichloromethane (6.6 kg), the filtrates were combined, concentrated under reduced pressure, the residue was dissolved in a mixed solvent of dichloromethane (5.5 kg) and ethanol (0.62 kg), the resulting solution was added to methyl tert-butyl ether (76 kg) under stirring, then n-heptane (70.5 kg) was added to the system, centrifuged under nitrogen protection, rinsed with n-heptane (5.8 kg), the solid was collected, dried under vacuum, and (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one was obtained, with a yield of 83.3%. 1 H NMR (400MHz, DMSO-d6) δ9.26 (s, 1H), 7.66-7.61 (m, 2H), 7.49 (t, J = 7.2Hz, 1H), 7.38-7.11 (m, 2H), 6.40 (s, 1H), 5.60-5.53 ( m,1H),3.01-2.98(m,4H),2.50-2.46(m,4H),2.23(s,3H),1.56-1.52(m,6H),1.25-1.05(m,4H); ESI-MS(m / z):487.0[M+H] + .

[0167] Example 2 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one hydrochloride

[0168] Ethanol (15.4 kg) was added to a 50 L reactor. Under stirring, (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (3.875 kg) was added. Ethyl hydrochloride solution (4.38 L) was slowly added at 10–40 °C. After the addition was complete, the reaction system was cooled to 20–30 °C. The reaction was stirred, and methyl tert-butyl ether (14.4 kg) was added dropwise to the reaction system. After the addition was complete, the mixture was stirred at 15-25°C for 24 h to crystallize. The mixture was centrifuged under nitrogen protection, washed with methyl tert-butyl ether (9.1 kg), and the solid was collected and dried under vacuum to obtain crude (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one hydrochloride. The crude product was added to ethanol (24 kg), heated to 70-80°C, and stirred until completely dissolved. The mixture was filtered while hot, and the filtrate was collected and cooled to 30-50°C. Methyl tert-butyl ether (22.6 kg) was slowly added. After the addition was complete, the mixture was cooled to 15-35°C, stirred to induce crystallization, centrifuged under nitrogen protection, and washed with methyl tert-butyl ether (19.3 kg). The solid was collected, dried under vacuum, and (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one hydrochloride was obtained, with a yield of 95.0%. 1 H NMR(400MHz,DMSO-d6)δ13.27(s,1H),11.70(s,1H),10.96(s,1H),10.38(s,1H),7 .85(t,J=7.2Hz,1H),7.61(t,J=7.2Hz,1H),7.41-7.13(m,2H),6.69(s,1H),5.61-5 .54(m,1H),3.61(d,J=9.6Hz,2H),3.47-3.41(m,2H),3.34-3.23(m,4H),2.78(s,3H ),1.78(d,J=6.4Hz,3H),1.55(s,3H),1.30-1.06(m,4H); ESI-MS(m / z):487.0[M+H] + Chloride ion content: 12.9%. Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with two molecules of hydrochloric acid.

[0169] Example 3 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one hydrobromide

[0170] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Hydrobromic acid (3.59 g, concentration 48%, 21.30 mmol) was added dropwise under stirring at room temperature. After the addition was complete, The reaction mixture was stirred at room temperature for 2 hours, and methyl tert-butyl ether (48.6 mL) was added dropwise. After the addition was complete, the mixture was stirred at room temperature overnight. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The resulting solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one hydrobromide, with a yield of 84.3%. 1 H NMR (400MHz, DMSO-d6) δ13.23(s,1H),10.05(s,1H),9.81(s,2H),7.77(t,J=7.2Hz,1 H),7.65(t,J=7.2Hz,1H),7.42-7.14(m,2H),6.73(s,1H),5.52-5.47(m,1H),3.65(d, J=5.6Hz,2H),3.53(d,J=12.4Hz,2H),3.39-3.30(m,2H),3.18-3.08(m,2H),2.87(s, 3H),1.74(d,J=4.0Hz,3H),1.57(s,3H),1.22-1.03(m,4H); ESI-MS(m / z):487.0[M+H] + Bromine ion content: 21.6%. Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.7 molecules of hydrobromic acid.

[0171] Example 4: Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one sulfate

[0172] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Sulfuric acid (2.13 g, concentration 98%, 21.28 mmol) was added dropwise under stirring at room temperature. After the addition was complete, the mixture was stirred. After 0.5 h, a large amount of solid precipitated. Methyl tert-butyl ether (48.6 mL) was added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one sulfate, with a yield of 92.0%. 1 H NMR (400MHz, DMSO-d6) δ13.21 (s, 1H), 9.69-9.66 (m, 3H), 7.73 (t, J = 7.6Hz, 1H), 7.6 5(t,J=7.6Hz,1H),7.41-7.14(m,2H),6.74(s,1H),5.49-5.42(m,1H),3.66(d,J=13 .6Hz,2H),3.53(d,J=15.6Hz,2H),3.37-3.32(m,2H),3.13-3.06(m,2H),2.88(s,3H ),1.72(d,J=6.4Hz,3H),1.54(s,3H),1.22-1.09(m,4H); ESI-MS(m / z):487.0[M+H] + Sulfate ion content: 18.3%. Structural identification showed that 1 molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.2 molecules of sulfate.

[0173] Example 5 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one phosphate

[0174] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Phosphoric acid (2.45 g, concentration 85%, 21.25 mmol) was added dropwise under stirring at room temperature. After stirring at room temperature for 2 hours, methyl tert-butyl ether (48.6 mL) was added dropwise, and the mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one phosphate, with a yield of 98.7%. 1 H NMR(400MHz,DMSO-d6)δ9.50(s,4H),9.30(s 1H),7.64(t,J=7.6Hz,1H),7.49(t,J=7.2Hz,1H),7.39-7.11(m,2H),6.47(s,1H),5.60-5.55(m,1H),3.24-3.18(m,4 H),3.14-3.10((m,4H),2.66(s,3H),1.56(d,J=4.8Hz,3H),1.54(s,3H),1.21-1.08(m,4H); ESI-MS(m / z):487.0[M+H] + Phosphate ion content: 27.8%. Structural identification revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one formed a salt with two molecules of phosphate.

[0175] Example 6 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one oxalate

[0176] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Oxalic acid (1.93 g, purity 99.36%, 21.27 mmol) was added under stirring at room temperature. After the addition was complete, the mixture was allowed to cool to room temperature. The mixture was stirred for 2 hours, and methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated out. The mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one oxalate, with a yield of 98.2%. 1 H NMR (400MHz, DMSO-d6) δ10.83(s,1H),9.40(s,1H),7.66(t,J=7.2Hz,1H),7.51(t,J=7.2Hz,1H),7.39-7.12(m,2H),6.52(s,1H),5 .59-5.54(m,1H),3.40-3.32(m,8H),2.80(s,3H),1.57(d,J=6.8Hz,3H),1.55(s,3H),1.20-1.08(m,4H); ESI-MS(m / z):487.0[M+H] + Oxalate ion content: 23.5%. Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.8 molecules of oxalate.

[0177] Example 7 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one citrate

[0178] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Citric acid monohydrate (4.50 g, purity 99.5%, 21.28 mmol) was added under stirring at room temperature. After the addition was complete, The mixture was stirred at room temperature for 2 hours, and methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated out. The mixture was stirred at room temperature overnight. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one citrate, with a yield of 92.3%. 1 H NMR (400MHz, DMSO-d6) δ10.83(s,1H),9.29(s,1H),7.66(t,J=7.2Hz,1H),7.50(t,J=7.2Hz,1H),7.39-7.12(m,2H),6.48(s,1H),5.60 -5.55(m,1H),3.25-3.13(m,8H),2.70-2.56(m,10H),1.56(d,J=4.4Hz,3H),1.54(s,3H),1.20-1.09(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.8 molecules of citric acid.

[0179] Example 8. Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one malate

[0180] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Malic acid (3.00 g, purity 95.0%, 21.27 mmol) was added with stirring at room temperature. After the addition was complete, the mixture was stirred at room temperature... The mixture was stirred at room temperature for 2 hours, and methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated out. The mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one malate, with a yield of 95.2%. 1 H NMR(400MHz,DMSO-d6)δ9.84(br s,1H),9.28(s,1H),7.64(t,J=7.6Hz,1H),7.49(t,J=7.2Hz,1H),7.39-7.1 1(m,2H),6.46(s,1H),5.60-5.54(m,1H),4.14(t,J=7.2Hz,1.8H),3.17-3.0 7(m,4H),2.99-2.94(m,4H),2.61-2.55(m,4.8H),2.43-2.37(m,1.8H),1.56 (t,J=3.6Hz,6H),1.54(s,3H),1.21-1.09(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.8 molecules of malic acid.

[0181] Example 9 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one maleate

[0182] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Maleic acid (2.49 g, purity 99.0%, 21.27 mmol) was added under stirring at room temperature. After the addition was complete, the mixture was allowed to cool to room temperature. The mixture was stirred for 0.5 h, and methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated out. The mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one maleate, with a yield of 91.7%. 1 H NMR(400MHz,DMSO-d6)δ9.45(s,1H),8.61(br s,1H),7.68(t,J=7.2Hz,1H),7.56(t,J=7.2Hz,1H),7.40-7.13(m,2H),6.61(s,1H),6.10(s,4H),5.55-5.50(m,1H ),3.58-3.30(m,8H),2.87(s,3H),1.62(d,J=6.4Hz,3H),1.55(s,3H),1.20-1.10(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with two molecules of maleic acid.

[0183] Example 10 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one fumarate

[0184] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Fumaric acid (2.48 g, purity 99.5%, 21.27 mmol) was added under stirring at room temperature. After the addition was complete, the mixture was stirred at room temperature for 0.5 h, and then methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated, and the mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The mixture was then dried under vacuum. The resulting solid was added to isopropanol (50 mL) at room temperature and stirred for 12 h. The mixture was then filtered under reduced pressure, and the filter cake was washed with isopropanol (10 mL). The resulting solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one fumarate, with a yield of 93.3%. 1 H NMR(400MHz, DMSO-d6)δ9.27(s,1H),7.64(t,J=7.6Hz,1H),7.49(t,J=7.2Hz,1H),7.39-7.11(m,2H),6.59(s,2.6H),6.44(s,1H),5.5 9-5.54(m,1H),3.11-3.07(m,4H),2.84-2.80(m,4H),2.46(s,3H),1.55(t,J=3.6Hz,6H),1.21-1.09(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.3 molecules of fumaric acid.

[0185] Example 11 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one succinate

[0186] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Succinic acid (2.52 g, purity 99.5%, 21.27 mmol) was added under stirring at room temperature. After the addition was complete, the mixture was allowed to cool to room temperature. The mixture was stirred for 2 hours, and methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated out. The mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one succinate, with a yield of 72.1%. 1 H NMR(400MHz, DMSO-d6)δ9.26(s,1H),7.63(t,J=7.2Hz,1H),7.49(t,J=7.2Hz,1H),7.39-7.11(m,2H),6.44(s,1H),5.59-5.54(m,1H) ,3.07-3.02(m,4H),2.62-2.58(m,4H),2.41(s,6H),2.23(s,3H),1.55(t,J=3.6Hz,6H),1.19-1.07(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.5 molecules of succinic acid.

[0187] Example 12 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one malonate

[0188] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Malonic acid (2.24 g, purity 99.0%, 21.27 mmol) was added under stirring at room temperature. After the addition was complete, the mixture was allowed to settle at room temperature. After stirring for 2 hours, methyl tert-butyl ether (48.6 mL) was added dropwise, resulting in the precipitation of a large amount of solid. The mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one malonate, with a yield of 85.3%. 1 H NMR(400MHz, DMSO-d6)δ9.30(s,1H),7.64(t,J=7.2Hz,1H),7.50(t,J=7.2Hz,1H),7.39-7.12(m,2H),6.48(s,1H),5.60-5.5 5(m,1H),3.23-3.16(m,8H),2.99(s,3.2H),2.73(s,3H),1.55(t,J=3.6Hz,6H),1.20-1.09(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with 1.6 molecules of malonic acid.

[0189] Example 13 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one methanesulfonate

[0190] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Methanesulfonic acid (2.07 g, purity 99.0%, 21.37 mmol) was added under stirring at room temperature. After the addition was complete, the mixture was allowed to settle at room temperature. The mixture was stirred for 2 hours, and methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated out. The mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazine-7(6H)-one methanesulfonate, with a yield of 89.8%. 1 H NMR (400MHz, DMSO-d6) δ13.24(s,1H),9.86(s,1H),9.75(d,J=7.6Hz,1H),9.68(s,1H),7.74(t, J=7.6Hz,1H),7.65(t,J=7.2Hz,1H),7.41-7.14(m,2H),6.74(s,1H),5.50-5.43(m,1H),3.66(d, J=5.6Hz,2H),3.52(d,J=12.4Hz,2H),3.37-3.29(m,2H),3.14-3.08(m,2H),2.88(d,J=4.4Hz,3H ),2.36(s,6H),1.72(d,J=6.8Hz,3H),1.57(s,3H),1.20-1.11(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with two molecules of methanesulfonic acid.

[0191] Example 14 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one benzenesulfonate

[0192] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, purity 96.85%, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Benzenesulfonic acid (3.43 g, purity 98.0%, 21.37 mmol) was added under stirring at room temperature. After the addition was complete, the mixture was allowed to settle at room temperature. The mixture was stirred for 2 hours, and methyl tert-butyl ether (48.6 mL) was added dropwise. A large amount of solid precipitated out. The mixture was stirred overnight at room temperature. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one benzenesulfonate, with a yield of 91.8%. 1 H NMR (400MHz, DMSO-d6) δ13.22(s,1H),9.64(s,1H),7.73(t,J=7.2Hz,1H),7.65(t,J=7.2 Hz,1H),7.61-7.58(m,4H),7.41-7.14(m,8H),6.73(s,1H),5.49-5.42(m,1H),3.65(d,J= 13.6Hz,2H),3.53(d,J=12.8Hz,2H),3.37-3.29(m,2H),3.10-3.03(m,2H),2.89(d,J=4.4 Hz,3H),1.71(d,J=6.8Hz,3H),1.55(s,3H),1.18-1.10(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with two molecules of benzenesulfonic acid.

[0193] Example 15 Synthesis of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one p-toluenesulfonate

[0194] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one (4.86 g, 96.85% purity, 9.674 mmol) was placed in a reaction flask, and isopropanol (48.6 mL) was added. Then, p-toluenesulfonic acid (3.78 g, 97.0% purity, 21.37 mmol) was added under stirring at room temperature. After adding l), the mixture was stirred at room temperature for 2 hours, and a large amount of solid precipitated. The mixture was stirred at room temperature overnight. The reaction system was filtered under reduced pressure, and the filter cake was washed with methyl tert-butyl ether (20 mL). The obtained solid was dried under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one p-toluenesulfonate, with a yield of 92.7%. 1 H NMR (400MHz, DMSO-d6) δ13.21(s,1H),9.70(m,2H),9.64(s,1H),7.73(t,J=7.6Hz,1H),7.64(t,J=7.2H z,1H),7.48(d,J=7.6Hz,3H),7.41-7.14(m,2H),7.11(d,J=8.0Hz,3H),7.10(s,2H),6.72(s,1H),5.48- 5.42(m,1H),3.66(d,J=13.6Hz,2H),3.53(d,J=12.0Hz,2H),3.37-3.29(m,2H),3.11-3.03(m,2H),2.8 9(s,3H),2.29(s,6H),1.70(d,J=6.4Hz,3H),1.55(s,3H),1.20-1.10(m,4H); ESI-MS(m / z):487.0[M+H] + Structural analysis revealed that one molecule of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one forms a salt with two molecules of p-toluenesulfonic acid.

[0195] Example 16 Preparation of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride crystal form A

[0196] (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride (3.8 kg) and ethanol (24 kg) were added, the mixture was heated to 70-80°C and stirred to dissolve, and then filtered while hot. The filtrate was cooled to 30-50°C, and methyl tert-butyl ether (22.6 kg) was slowly added. After addition, the mixture was cooled to 15–35 °C, stirred to allow crystals to precipitate for 24 h, centrifuged under nitrogen protection, and washed with methyl tert-butyl ether (19.3 kg). The resulting solid was dried under vacuum to give product A, (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride, with a yield of 95.0%.

[0197] The XRD pattern of crystal form A is shown in Figure 1, the DSC pattern in Figure 4, and the TGA pattern in Figure 6. The XRD pattern analysis data are shown in Table 1. The TGA and DSC patterns show that the sample loses approximately 3.16% of its weight when heated to 110℃ and begins to decompose when heated to around 200℃.

[0198] Table 1. XRPD pattern analysis data for crystal form A

[0199] Example 17 Preparation of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride crystal form B

[0200] Add (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride crystal form A (30 g) and ethanol (240 mL) to a 500 mL reaction flask. Heat to reflux and stir to dissolve. Filter while hot. Cool the filtrate to 15–35 °C and stir to crystallize for 96 h. Filter. Wash the filter cake with ethanol (20 mL). Dry the obtained solid under vacuum to give (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride crystal form B, yield 51.7%.

[0201] The XRPD pattern of crystal form B is shown in Figure 2, the DSC pattern in Figure 5, and the TGA pattern in Figure 7. The XRPD pattern analysis data are shown in Table 2. The TGA and DSC patterns show that the sample loses approximately 5.60% of its weight when heated to 150℃ and begins to decompose when heated to around 220℃.

[0202] Table 2. XRPD pattern analysis data for crystal form B.

[0203] Example 18: Preparation of amorphous (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride

[0204] 15.01 g of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride crystal form A was added to a 500 mL reaction flask, and purified water (300 mL) was added. The mixture was stirred and dissolved, filtered, and the filtrate was freeze-dried to obtain the amorphous product of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one dihydrochloride, with a yield of 100%.

[0205] The amorphous XRPD pattern is shown in Figure 3.

[0206] The following experimental examples demonstrate the beneficial effects of the present invention.

[0207] Experiment 1: Solubility Test of Different Salt Forms

[0208] In this experiment, the solubility of different salts of compound ((R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one in pure water was determined by high performance liquid chromatography with external standard method.

[0209] 1.1 Preparation of external standard samples

[0210] Accurately weigh approximately 20 mg of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one hydrochloride, dissolve and dilute with diluent (acetonitrile / water = 25 / 75) to prepare a solution containing approximately 0.2 mg per mL, which will serve as the external standard reference solution.

[0211] 1.2 Preparation of Solubility Test Samples

[0212] Each salt form was investigated independently: 1 mL of pure water was accurately measured and placed in a 2 mL sample bottle, and (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one free base, hydrochloride, hydrobromide, sulfate, phosphate, oxalate, citrate, malate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, maleate, fumarate, succinate, and malonate sample powder were continuously added to the sample bottle until the solid powder no longer dissolved, thus preparing a supersaturated solution of the compound.

[0213] The sample was shaken at 25℃ and 200 rpm. 100 μl of the sample was taken after 22 h, centrifuged (5000 rpm) for 5 min, and diluted by different factors to prepare a solubility test solution.

[0214] 1.3 Sample Testing

[0215] The samples were analyzed by high-performance liquid chromatography (HPLC). The solubility of the free base and different salt forms of (R)-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-6-(1-methylcyclopropyl)-1-(4-methylpiperazin-1-yl)pyrido[3,4-d]pyridazin-7(6H)-one in pure water was calculated by comparing the integrated peak area with that of a known concentration external standard. The solubility test results are shown in Table 3.

[0216] 1.4 High Performance Liquid Chromatography Method

[0217] Column: Welch Xtimate C18 4.6*150mm, 3μm

[0218] Flow rate: 1.5 mL / min

[0219] Detection wavelength: 261nm

[0220] Column temperature: 30℃

[0221] Injection volume: 10 μl

[0222] Analysis duration: 15 min

[0223] Mobile phase: 50 mmol / L ammonium bicarbonate (adjusted to pH 8.2 with diethylamine) - acetonitrile = 62:38

[0224] Diluent: Acetonitrile-water (25:75)

[0225] Gradient procedure: isotropic method

[0226] Table 3. Solubility test results of different salt types

[0227] As shown in Table 3, the solubility of the free alkali in pure water is only 0.003 mg / mL. After salt formation, the solubility of each salt form is significantly better than that of the free alkali, with all reaching a level much greater than 10 mg / mL in pure water, among which the hydrochloride salt has the highest solubility.

[0228] Experiment 2: Stability Tests of Different Salt Forms

[0229] 2.1 Influencing Factors and Conditions:

[0230] High temperature: Take an appropriate amount of sample, place it in a clean flat weighing bottle, and place it in an open drying oven at 60℃±2℃. Take samples for testing at 0 days, 7 days, and 14 days.

[0231] Light exposure: Take an appropriate amount of sample and place it in a clean, transparent, flat weighing bottle. Place the sample in a high-intensity light stability test chamber (visible light 4500 lux ± 500 lux, ultraviolet light 84 μw / cm²). 2 The samples were placed in an open container and tested at 0, 7 and 14 days.

[0232] Impurity content was calculated using HPLC with peak area normalization. The variations in total impurity content for each salt type are shown in Tables 4 and 5.

[0233] 2.2 High Performance Liquid Chromatography (HPLC) Conditions:

[0234] Column: Waters Xbridge C18 4.6mm*150mm 5μm

[0235] Flow rate: 1.0 mL / min

[0236] Wavelength: 261nm

[0237] Column temperature: 30℃

[0238] Sample injection chamber temperature: 15℃

[0239] Injection volume: 10 μl

[0240] Runtime: 55 min

[0241] Mobile phase A: 10 mmol / L potassium dihydrogen phosphate (pH adjusted to 7.2 with potassium hydroxide solution)

[0242] Mobile phase B: Acetonitrile

[0243] Diluent: Water-acetonitrile (80:20)

[0244] Gradient procedure:

[0245] Table 4. Total Impurity Increase under High Temperature Conditions

[0246] As shown in Table 4, when stored at high temperature for a long time (14 days), the total impurity content of hydrochloride and oxalate increases the least, indicating good stability and promising medicinal prospects.

[0247] Table 5. Increase in total impurities under illumination conditions

[0248] As can be seen from Table 5, when placed under light conditions for a long time, the total impurity content of each salt type changes less than that of the free alkali. Among them, the total impurity content of hydrochloride, hydrobromide, sulfate and citrate increases by only about 1%, showing good stability and promising medicinal prospects.

[0249] Experiment 3: Pharmacokinetic Study of Rats with Different Salt Forms

[0250] Female SD rats, weighing 200-250g, were used and fasted overnight. Three rats were used in each group and administered the test compound suspension or solution (0.5% w / w methylcellulose aqueous solution) orally (PO). The plasma concentration of the test compound in the rats was determined by LC-MS / MS, and the main pharmacokinetic parameters were calculated to evaluate the in vivo pharmacokinetic behavior. Whole blood was collected before PO administration and at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. The plasma was collected in K2-EDTA anticoagulant tubes, centrifuged within 10 min (4℃), and stored at -80℃ for analysis. For sample processing, protein precipitation was performed using an appropriate amount of methanol or acetonitrile containing an internal standard. After vortexing and centrifugation, the supernatant was injected for LC-MS / MS analysis. The main pharmacokinetic parameters were analyzed using a non-compartmental model in WinNonLin 8.3 software. The main pharmacokinetic parameters are shown in Table 6.

[0251] Table 6. Pharmacokinetic parameters of rats

[0252] In vivo pharmacokinetic studies in rats showed that the exposure levels of the free base and its different salt forms varied considerably. As shown in Table 6, compared to the free base, the in vivo drug exposure levels of hydrochloride, oxalate, and p-toluenesulfonate were significantly increased, exhibiting superior pharmacokinetic properties and better drug-likeness.

[0253] Based on the results of Experiments 1 to 3, the hydrochloride salt has a solubility greater than 10.0 mg / mL in pure water, meeting the general requirements for formulation development; it exhibits good stability under high temperature and light conditions, which is beneficial for formulation process development and storage; and its exposure level in rats is relatively high. In comparison, the hydrochloride salt is the salt form with superior overall performance.

[0254] Experimental Example 4: Equilibrium Solubility Test of Different Crystal Forms

[0255] In this experiment, the equilibrium solubility of the compound in pure water was determined by high performance liquid chromatography with external standard method. The results are shown in Table 7.

[0256] 4.1 Preparation of equilibrium solubility samples

[0257] Take appropriate amounts of samples of different crystal forms and place them in 2 mL sample vials respectively. Add an appropriate amount of water to make the solution supersaturated. Place the prepared saturated solution on a constant temperature shaker at 37℃ and 200 rpm. Take 100 μl samples at 0.5 h, 2 h, and 24 h respectively, centrifuge (12000 rpm) for 10 min, take the supernatant and dilute it to different concentrations, and shake well to obtain the final solution.

[0258] Accurately weigh the reference standard, dissolve it in a solvent to prepare a 0.2 mg / mL solution, and determine the content of each sample using the external standard method.

[0259] 4.2 High Performance Liquid Chromatography Method

[0260] Column: Welch Xtimate C18 4.6*150mm, 3μm

[0261] Flow rate: 1.5 mL / min

[0262] Detection wavelength: 261nm

[0263] Column temperature: 30℃

[0264] Injection volume: 10 μl

[0265] Analysis duration: 15 min

[0266] Mobile phase: 50 mmol / L ammonium bicarbonate (adjusted to pH 8.2 with diethylamine): acetonitrile = 62:38

[0267] Diluent: Acetonitrile-water (25:75)

[0268] Gradient procedure: isotropic method

[0269] Table 7. Results of equilibrium solubility tests for different crystal forms

[0270] As shown in Table 7, the solubility of hydrochloride crystal forms A and B in pure water is greater than 500 mg / mL, which is higher than the solubility of amorphous hydrochloride.

[0271] Experimental Example 5: Stability Tests of Different Crystal Forms

[0272] 5.1 Influencing Factors and Conditions:

[0273] High temperature: Take an appropriate amount of sample, place it in a clean flat weighing bottle, and place it in an open drying oven at 60℃±2℃. Take samples for testing at 0 days, 14 days, and 30 days.

[0274] High humidity: Take an appropriate amount of sample and place it in a clean, flat weighing bottle. Place the bottle open in a desiccator containing saturated potassium nitrate solution (75%RH±5%RH). Take samples for testing at 0 days, 14 days, and 30 days.

[0275] Humid heat: Take an appropriate amount of sample, place it in a clean flat weighing bottle, and place it open in a drug stability test chamber (40℃±2℃ / 75%RH±5%RH). Take samples for testing at 0 days, 14 days, and 30 days.

[0276] Impurity content was calculated using HPLC with peak area normalization. The variation of total impurity content for each crystal form is shown in Table 8.

[0277] 5.2 High Performance Liquid Chromatography (HPLC) Conditions:

[0278] Column: Welch Xtimate C18 4.6*150mm, 3μm

[0279] Flow rate: 1.0 mL / min

[0280] Wavelengths: 261nm, 210nm

[0281] Column temperature: 30℃

[0282] Sample injection chamber temperature: 15℃

[0283] Injection volume: 10 μl

[0284] Running time: 60min

[0285] Mobile phase A: 50 mmol / L ammonium bicarbonate (adjusted to pH 8.2 with ammonia)

[0286] Mobile phase B: Acetonitrile

[0287] Diluent: Water-acetonitrile (25:75)

[0288] Gradient procedure:

[0289] Table 8 Results of Total Impurity Increase for Different Crystal Forms

[0290] As shown in Table 8, the total impurity content of crystal forms A and B did not change significantly under high temperature, high humidity and humid heat conditions, indicating that they have good stability and are better than amorphous forms.

[0291] Experimental Example 6: TGA / DSC Testing of Different Crystal Forms

[0292] Instrument Model: TGA / DSC 2 Thermogravimetric Differential Scanning Calorimeter

[0293] Test method: Take a sample (about 1-10 mg) and place it in a TGA / DSC test dish. Under N2 conditions of 50 mL / min, heat the sample from 40 °C to 350 °C at a heating rate of 10 °C / min.

[0294] The DSC / TGA spectra of crystalline forms A, B, and the amorphous form are shown in Figures 4-7. The spectra show that crystalline forms A and B are both hydrates. The TGA spectrum of crystalline form A shows weight loss within the temperature range of 40℃ to 110℃, indicating the presence of adsorbed water; the TGA spectrum of crystalline form B shows weight loss within the temperature range of 40℃ to 150℃, also indicating the presence of adsorbed water. Combined with the DSC / TGA spectra, it can be seen that crystalline form A begins to decompose at around 200℃, and crystalline form B begins to decompose at around 220℃, with increasing ambient temperature. No other obvious endothermic or exothermic peaks were observed before decomposition in either crystalline form, indicating that no crystal transformation occurred, demonstrating the good stability of crystalline forms A and B.

[0295] Experimental Example 7: Hygroscopicity Test of Different Crystal Forms

[0296] Take 1g of each of the different crystal forms, accurately weigh it (m2), and place it in a stoppered glass weighing bottle (accurately weighed (m1)) at (25℃±1℃, relative humidity 80%±2%) for 24 hours. After leaving it open for 24 hours at (25℃±1℃, relative humidity 80%±2%), accurately weigh it (m3) and calculate the percentage of moisture absorption weight gain according to the following formula.

[0297] The test results are as follows:

[0298] Table 9. Hygroscopicity test results for different crystal forms

[0299] As shown in Table 9, the moisture absorption weight gain of crystal forms A and B is significantly lower than that of amorphous forms, indicating that crystal forms A and B have better moisture absorption stability than amorphous forms.

[0300] Experiment Example 8: Bioactivity Experiment

[0301] (1) Assay of SOS1 inhibitory activity

[0302] The effect of SOS1 inhibitors on the interaction between SOS1 and KRAS proteins was assessed using homogeneous time-resolved fluorescence (HTRF) to evaluate their inhibitory level on SOS1. The protein and assay reagents used were the KRAS-G12D / SOS1 binding ASSAY KIT (Cisbio). First, a 2 mM stock solution of the test compound was diluted 20-fold (100 μM) with Diluent reagent, followed by a 5-fold serial dilution with Diluent reagent (5% DMSO) to obtain eight working solutions of the test compound. 4 μL of tag2-KRAS was added to each well of a 384-well plate. G12D Protein (containing 50 μM GTP), 2 μL of the test compound, and 4 μL of tag1-SOS1 protein were added to each well in duplicate and incubated at room temperature for 15 min. Anti-tag1-Tb was then added to each well sequentially. 3+ 5 μL each of the working solution and Anti-tag2-XL665 working solution were incubated at 4 °C for 3 h. The 384-well plate was then placed on a multi-mode microplate reader, with the excitation wavelength set to 337 nm. Readings at 620 nm and 665 nm were recorded. Data are presented as the ratio of the 665 nm signal value to the 620 nm signal value per well, i.e., Ratio = 10. 4 ×665nm signal value / 620nm signal value. The suppression rate is calculated using the ratio value.

[0303] % Inhibition rate = [(Ratio 阴性 -Ratio 化合物 ) / (Ratio 阴性 -Ratio Blank )]×100

[0304] IC 50 The inhibition rate was calculated using GraphPad Prism software. Two replicates were performed for each compound. The test data for the compounds in the embodiments of this invention are as follows:

[0305] Table 10. Inhibitory activity of compounds from the embodiments of the present invention against SOS1.

[0306] As shown in Table 10, the inhibitory activity of different salt forms and crystal forms of this compound against SOS1 is similar to that of the free base.

[0307] (2) Determination of inhibitory activity against NCI-H358 cell proliferation

[0308] via CellTiter-Glo The number of viable cells was measured using reagents to assess the inhibitory effect of the compound on cell proliferation. Non-small cell lung cancer cells (NCI-H358, Nanjing Kebai) in the exponential growth phase were digested with trypsin-EDTA and seeded into 96-well ultra-low adsorption microplates at a density of 2000 cells / well (100 μL per well). The cells were cultured overnight at 37°C and 5% CO2 to form microspheres. The test compound was serially diluted 5-fold with DMSO to obtain eight concentration gradient dilutions. These were then diluted with RPMI-1640 (10% FBS) cell culture medium to obtain the working solution (2×), and 100 μL was added to the cell supernatant per well. The cells were cultured for another 7 days at 37°C and 5% CO2. The plates were then removed, and 100 μL of CellTiter-Glo was added to each well. After lysing the reagents at room temperature for 10-30 minutes, aspirate 70 μL of the lysis buffer and transfer it to a white, opaque 384-well plate. Place the 384-well plate on a multi-plate reader to record the Luminescence value (RLU).

[0309] The inhibition rate is calculated using the following formula: Inhibition rate (%) = {[(RLU)} 阴性对照 -RLU 空白 )-(RLU 受试化合物 -RLU 空白 )] / (RLU 阴性对照 -RLU 空白 )}×100%

[0310] Note: The negative control group was the group without inhibitors, and the blank group was the group without cells.

[0311] IC 50 The inhibition rate was calculated using GraphPad Prism software. The test results of the inhibitory activity of the compounds in this invention against the proliferation of NCI-H358 cells are as follows:

[0312] Table 11 Inhibitory activity of compounds from the embodiments of the present invention against NCI-H358 cell proliferation

[0313] As shown in Table 11, the cell proliferation inhibition activities of different salt forms and crystal forms of this compound on NCI-H358 are similar to those of the free base.

[0314] Experimental Example 9: Determination of Hepatic Microsomal Metabolic Stability

[0315] Metabolic stability affects the clearance rate, half-life, and oral bioavailability of compounds in the body, and is one of the most important ADME (adverse drug reaction) properties of compounds. Liver microsomal stability assays are a commonly used method for studying metabolic stability. This invention uses rat liver microsomal assays to investigate the metabolic stability of compounds.

[0316] The test method is as follows:

[0317] a. Solution preparation

[0318] Preparation of stock solution: Take an appropriate amount of the test compound and testosterone, prepare a 1 mM stock solution with DMSO, and store in a refrigerator at 4°C.

[0319] Preparation of compound working solution: Take 1 mM of the compound stock solution and dilute it with acetonitrile-water (v:v = 1:1) to 100 μM.

[0320] Preparation of NADPH working solution: Weigh an appropriate amount of NADPH and prepare a 4.0 mM NADPH working solution with 16 mM MgCl2.

[0321] Preparation of working solution for rat liver microsomes: Take an appropriate amount of rat liver microsomes and dilute them with PBS to prepare a working solution of 1 mg / ml for liver microsomes.

[0322] b. Sample incubation

[0323] Add 48 μL PBS, 100 μL rat liver microsomal working solution, and 2 μL working solution of the test compound sequentially to the incubation system, and mix thoroughly. After pre-incubation at 37°C for 5 min, add 50 μL NADPH to start the reaction. After the corresponding incubation time points, add an appropriate amount of ice-cold acetonitrile containing internal standard to terminate the reaction, vortex to mix, centrifuge to collect the supernatant, and inject LC-MS / MS to detect the residual amount of the compound.

[0324] c. Data Processing

[0325] Use software such as Excel to process the data according to the following formula:

[0326] CL (liver) =(CL) int(liver) ×f u ×Q h ) / (CL int(liver )×f u +Q h )

[0327] default f u The free fraction in the blood is equal to 1.

[0328] The stability test results of the compounds in some embodiments of the present invention on rat liver microsomes are as follows:

[0329] Table 12 Results of metabolic stability tests on compounds from the embodiments of the present invention

[0330] MRTX0902 is in Phase II clinical trials and is being developed by Bristol-Myers Squibb. As shown in Table 12, the compound of this invention exhibits better metabolic stability and better drug-like properties compared to compound MRTX0902.

[0331] Experimental Example 10: Assay of Hepatic Drug Metabolizing Enzyme Inhibitory Activity

[0332] The liver is the primary site of drug metabolism, and the main component of the mixed-function oxidase system in the liver is the CYP450 enzyme. This enzyme system causes interactions with most drugs in clinical practice, leading to an increased incidence of adverse drug reactions. CYP450 is a superfamily of enzymes composed of many isoenzymes, among which CYP3A4 is the major metabolic enzyme involved in the metabolism of nearly half of the drugs used clinically. Similarly, CYP2C9 and CYP2C19 are also important metabolic enzymes involved in the metabolism of various drugs clinically. Therefore, early testing of the inhibitory activity of compounds on CYP450 enzymes can assess the risk of drug-drug interactions and improve medication safety.

[0333] In this experiment, human liver microsomes were used as the CYPP450 enzyme source. Specific probe substrates for each CYP isoenzyme (CYP3A4 used two substrates, midazolam and testosterone, denoted as CYP3A4-M and CYP3A4-T, respectively) were incubated with a series of concentrations of the test compound in the presence of the cofactor NADPH. The amount of metabolites generated from the probe substrates in the incubation system was determined using LC-MS / MS, and the IC50 values ​​of the test compounds for each CYP450 enzyme isoform were calculated. 50 The inhibitory effect of the probe on various CYP450 enzyme isoforms was evaluated. In the experiment, 49 μL of PBS, 50 μL of probe substrate, and 50 μL of human liver microsomal working solution were added sequentially to the incubation system, followed by 1 μL of working solution of each concentration of the test compound. The mixture was thoroughly mixed. After pre-incubation at 37°C for 5 min, 50 μL of NADPH was added to initiate the reaction. After the appropriate incubation time, an appropriate amount of ice-cold acetonitrile containing the internal standard was added to terminate the reaction. The mixture was vortexed, centrifuged, and the supernatant was collected. The amount of metabolites generated by the probe substrate was detected by LC-MS / MS. The percentage of residual enzyme activity (characterized by the amount of metabolites generated) at 0 concentration was taken as 100%, and the IC50 value was calculated for different concentrations of the test compound. 50 The remaining enzyme activity was calculated using Graphpad Prism software.

[0334] Experimental results show that the inhibitory activity of the compounds of this invention against CYP2C9, CYP2C19, CYP3A4-M, and CYP3A4-T is significantly weaker than that of the reference compound MRTX0902, which is beneficial in reducing the risk of drug-drug interactions. For example, Table 13 below lists the test data of two typical compounds of this invention and the reference compound MRTX0902:

[0335] Table 13 Drug-metabolizing enzyme inhibitory activities of the compounds in the embodiments of the present invention

[0336] It should be noted that the specific features, structures, materials, or characteristics described in this specification may be combined in any suitable manner in one or more embodiments. Furthermore, those skilled in the art can combine and integrate the different embodiments described in this specification and the features of those embodiments without contradiction.

Claims

1. The hydrochloride, hydrobromide, sulfate, phosphate, oxalate, citrate, malate, maleate, fumarate, succinate, malonate, methanesulfonate, benzenesulfonate, and p-toluenesulfonate of the compounds shown in Formula I:

2. The hydrochloride salt of the compound shown in Formula I, wherein the molar ratio of the compound shown in Formula I to hydrochloric acid is 1:

2.

3. The amorphous form of the hydrochloride salt according to claim 1 or 2.

4. The amorphous form as described in claim 3, characterized in that: Its X-ray powder diffraction pattern is shown in Figure 3.

5. The crystalline form of the hydrochloride salt according to claim 1 or 2.

6. The crystalline form as described in claim 5, characterized in that: Selected from anhydrous crystal form, hydrate crystal form, and solvate crystal form.

7. The crystalline form as described in claim 5 or 6, characterized in that: The crystalline form is crystal form A, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.68±0.2°, 11.01±0.2°, 14.64±0.2°, 18.70±0.2° and 24.24±0.2°.

8. The crystalline form as described in claim 7, characterized in that: The X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 11.01±0.2°, 12.70±0.2°, 14.64±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, and 24.24±0.2°.

9. The crystalline form as described in claim 8, characterized in that: The X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 11.01±0.2°, 12.70±0.2°, 14.64±0.2°, 18.25±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, 24.24±0.2°, and 24.65±0.2°.

10. The crystalline form as described in any one of claims 7 to 9, characterized in that: The X-ray powder diffraction pattern of crystal form A also has characteristic diffraction peaks at the following 1 to 10 2θ angles: 10.40±0.2°, 12.26±0.2°, 13.24±0.2°, 13.75±0.2°, 15.21±0.2°, 16.44±0.2°, 22.16±0.2°, 25.45±0.2°, 27.85±0.2°, and 29.05±0.2°.

11. The crystalline form as described in claim 10, characterized in that: The X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 10.40±0.2°, 11.01±0.2°, 12.26±0.2°, 12.70±0.2°, 13.24±0.2°, 13.75±0.2°, 14.64±0.2°, 18.25±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, 24.24±0.2°, 24.65±0.2°, 25.45±0.2°, and 29.05±0.2°.

12. The crystalline form as described in claim 11, characterized in that: The X-ray powder diffraction pattern of crystal form A exhibits characteristic diffraction peaks at the following 2θ angles: 4.05±0.2°, 8.15±0.2°, 8.68±0.2°, 10.40±0.2°, 11.01±0.2°, 12.26±0.2°, 12.70±0.2°, 13.24±0.2°, 13.75±0.2°, 14.64±0.2°, and 15.21±0.2°. 2°, 16.44±0.2°, 17.37±0.2°, 18.25±0.2°, 18.70±0.2°, 19.81±0.2°, 20.95±0.2°, 22.16±0.2°, 23.04±0.2°, 24.24±0.2°, 24.65±0.2°, 25.45±0.2°, 27.85±0.2° and 29.05±0.2°.

13. The crystalline form as described in claim 5 or 6, characterized in that: The crystal form is crystal form A, and its X-ray powder diffraction pattern is shown in Figure 1.

14. The crystalline form as described in claim 5 or 6, characterized in that: The crystalline form is crystal type B, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 9.38±0.2°, 14.64±0.2°, 23.03±0.2° and 25.80±0.2°.

15. The crystalline form as described in claim 14, characterized in that: The X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 9.38±0.2°, 14.64±0.2°, 18.34±0.2°, 23.03±0.2° and 25.80±0.2°.

16. The crystalline form as described in claim 15, characterized in that: The X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 9.38±0.2°, 14.64±0.2°, 17.43±0.2°, 18.34±0.2°, 19.73±0.2°, 23.03±0.2°, 24.64±0.2°, 25.80±0.2°, and 29.90±0.2°.

17. The crystalline form as described in any one of claims 14 to 16, characterized in that: The X-ray powder diffraction pattern of crystal form B also exhibits characteristic diffraction peaks at the following 1 to 11 2θ angles: 8.29±0.2°, 10.99±0.2°, 11.84±0.2°, 12.25±0.2°, 12.83±0.2°, 15.05±0.2°, 15.45±0.2°, 16.60±0.2°, 22.45±0.2°, 29.20±0.2°, and 30.50±0.2°.

18. The crystalline form as described in claim 17, characterized in that: The X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 8.29±0.2°, 9.38±0.2°, 10.99±0.2°, 12.83±0.2°, 14.64±0.2°, 16.60±0.2°, 17.43±0.2°, 18.34±0.2°, 19.73±0.2°, 20.75±0.2°, 23.03±0.2°, 24.64±0.2°, 25.80±0.2°, 29.20±0.2°, and 29.90±0.2°.

19. The crystalline form as described in claim 18, characterized in that: The X-ray powder diffraction pattern of crystal form B exhibits characteristic diffraction peaks at the following 2θ angles: 4.13±0.2°, 8.29±0.2°, 9.38±0.2°, 10.99±0.2°, 11.84±0.2°, 12.83±0.2°, 14.64±0.2°, 15.05±0.2°, 15.45±0.2°, 16.60±0.2°, 17.43±0.2°, and 17.75±0.2°. 2°, 18.34±0.2°, 18.64±0.2°, 19.55±0.2°, 19.73±0.2°, 20.75±0.2°, 21.05±0.2°, 21.33±0.2°, 23.03±0.2°, 24.64±0.2°, 25.80±0.2°, 26.45±0.2°, 28.25±0.2°, 29.20±0.2°, 29.90±0.2°.

20. The crystalline form as described in claim 5 or 6, characterized in that: The crystal form is crystal type B, and its X-ray powder diffraction pattern is shown in Figure 2.

21. The amorphous form as described in claim 4 or the crystalline form as described in any one of claims 7 to 20, characterized in that: The X-ray powder diffraction pattern was obtained under Cu Kα radiation conditions.

22. The method for preparing the crystalline form according to any one of claims 5 to 13, characterized in that: Recrystallization of the hydrochloride salt of the compound shown in Formula I with ethanol and methyl tert-butyl ether yields crystal form A.

23. The method for preparing the crystalline form according to any one of claims 5, 6, 14-20, characterized in that: Recrystallization of the hydrochloride crystal form A of the compound shown in Formula I with ethanol yields crystal form B.

24. A pharmaceutical composition, characterized by: It contains the salt as described in claim 1 or 2, the amorphous form as described in claim 3 or 4, or the crystalline form as described in any one of claims 5 to 21, and pharmaceutically acceptable excipients or auxiliary ingredients.

25. Use of the salt of claim 1 or 2, the amorphous form of claim 3 or 4, the crystalline form of any one of claims 5 to 21, or the pharmaceutical composition of claim 24 in the preparation of an SOS1 inhibitor.

26. Use of the salt of claim 1 or 2, the amorphous form of claim 3 or 4, the crystalline form of any one of claims 5 to 21, or the pharmaceutical composition of claim 24 in the preparation of a medicament for treating SOS1-mediated diseases.

27. The use as described in claim 26, characterized in that: The disease is selected from at least one of cancer and pathogenic skin diseases.

28. The use as described in claim 27, characterized in that: The cancers mentioned are selected from non-small cell lung cancer, lung cancer, pancreatic cancer, ovarian cancer, bladder cancer, prostate cancer, chronic myeloid leukemia, colorectal cancer, brain cancer, liver cancer, kidney cancer, stomach cancer, and breast cancer. The pathogenic skin rashes mentioned are selected from Noonan syndrome, cardiofacial skin syndrome, and type I hereditary gingival fibromatosis.

29. Use of the salt of claim 1 or 2, the amorphous form of claim 3 or 4, the crystalline form of any one of claims 5 to 21, or the pharmaceutical composition of claim 24 in the preparation of a medicament for treating diseases caused by overexpression of the SOS1 protein.

30. Use of the salt of claim 1 or 2, the amorphous form of claim 3 or 4, the crystalline form of any one of claims 5 to 21, or the pharmaceutical composition of claim 24 in the preparation of a medicament for treating diseases caused by SOS1 protein overexpression.

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