TW202428573A - Salt and crystal form of phosphonyl derivative and use thereof in medicine - Google Patents

Abstract

Provided in the present invention are a pharmaceutically acceptable salt of a compound as represented by formula (I) or a stereoisomer thereof, a solvate and/or a crystal form thereof, a preparation method therefor, a pharmaceutical composition thereof, and the use thereof in medicine.

Description

Salts and crystal forms of phosphonyl derivatives and their use in medicine

The present invention relates to the field of medicine, and more specifically, to a salt and a crystal form of a compound represented by formula (I) and a preparation method thereof, as well as the use of the salt and a crystal form thereof in a pharmaceutical composition and in medicine.

Epidermal growth factor receptor (EGFR) is a transmembrane protein tyrosine kinase that acts as a receptor for EGF family members to trigger the EGFR signaling pathway in human epithelial cells, thereby regulating cell proliferation, invasion, metastasis, apoptosis and angiogenesis (Nat. Rev. Cancer, 2007, 7, 169−181; Expert Opin. Ther. Targets, 2012, 16, 15−31.). Overexpression, mutation or amplification of the EGFR gene in the human body leads to abnormal increase in EGFR activity, which can lead to the occurrence of many malignant tumors such as esophageal cancer, glioblastoma, anal cancer, head and neck epithelial cancer, breast cancer, lung cancer, especially non-small cell lung cancer (NSCLC) (Cells, 2019, 8, 350−361.). PROTAC (proteolysis targeting chimera) molecules are a class of bifunctional compounds that can simultaneously bind to target proteins and E3 ubiquitin ligases. Such compounds can be recognized by the proteasome of cells, causing the degradation of target proteins, and can effectively reduce the content of target proteins in cells. By introducing ligands that can bind to different target proteins into PROTAC molecules, PROTAC technology can be applied to the treatment of various diseases. This technology has also received widespread attention in recent years (ACS Chem. Biol. 2017,12, 892−898; Drug Discovery Today Technol. 2019, 31, 15−27.).

PCT/CN2022/090243 describes a compound represented by formula (I), which is a Protacs small molecule with good EGFR inhibition and degradation activity. (I).

The purpose of the present invention is to provide a pharmaceutically acceptable salt of a compound represented by formula (I), a crystal form thereof, and a preparation method thereof, a pharmaceutical composition thereof, and use thereof in preparing drugs for EGFR-related diseases such as cancer diseases.

The advantages of the compound represented by formula (I) or its stereoisomer, the pharmaceutically acceptable salt and the solvate thereof and the crystalline or amorphous form of the compound represented by formula (I) include but are not limited to easy processing and crystallization, convenient handling, easy purification, easy industrialization, good fluidity, easy micronization, high solubility, good pharmacokinetic properties and good stability, and are suitable for the preparation of pharmaceutical preparations.

The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt of its stereoisomer and a solvate thereof. (I)

In some embodiments, the pharmaceutically acceptable salt is selected from maleate, 2-naphthalenesulfonate, 1,5-naphthalene disulfonate, fumarate, hydrohalide (preferably hydrobromide and hydrochloride), sulfate, phosphate, L-tartrate, citrate, L-malate, hippurate, D-glucuronide, glycolate, mucate, succinate, lactate, orotic acid. salt, pamoate, glycine, alanine, arginine, cinnamate, benzoate, benzenesulfonate, p-toluenesulfonate, acetate, propionate, valerate, triphenylacetate, L-proline, ferulate, 2-hydroxyethanesulfonate, mandelate, nitrate, methanesulfonate, malonate, gentianate, salicylate, oxalate, or glutarate;

In some embodiments, the pharmaceutically acceptable salt is selected from benzenesulfonate, L-applesulfate, phosphate, sulfate, p-toluenesulfonate, hydrochloride, maleate, 2-naphthalenesulfonate, hydrobromide, methanesulfonate, citrate, mandelate, lactobionate, succinate, salicylate, 1,5-naphthalene disulfonate, fumarate, nicotinate, hippurate and oxalate;

In some embodiments, the pharmaceutically acceptable salt is selected from methanesulfonate, maleate, 2-naphthalenesulfonate, and oxalate;

In some embodiments, the pharmaceutically acceptable salt is selected from a methanesulfonate salt;

In some embodiments, the molar ratio of the compound represented by formula (I): the pharmaceutically acceptable salt is 1:0.5 to 1:3.5;

In some embodiments, the molar ratio of the compound represented by formula (I): the pharmaceutically acceptable salt is 1:1, 1:2, or 1:3;

In some embodiments, the pharmaceutically acceptable salt is selected from methanesulfonate salts, and the molar ratio of the compound represented by formula (I): methanesulfonic acid is 1:1, 1:2, 1:3;

In some embodiments, the pharmaceutically acceptable salt is selected from methanesulfonate salts, and the molar ratio of the compound represented by formula (I): methanesulfonic acid is 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from maleic acid salts, and the molar ratio of the compound represented by formula (I): maleic acid is 1:2 or 1:1;

In some embodiments, the pharmaceutically acceptable salt is selected from 2-naphthalenesulfonic acid salts, and the molar ratio of the compound represented by formula (I): 2-naphthalenesulfonic acid is 1:1, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from oxalate salts, and the molar ratio of the compound represented by formula (I): oxalic acid is 1:1;

In some embodiments, the pharmaceutically acceptable salt is selected from benzenesulfonate salts, and the molar ratio of the compound represented by formula (I): benzenesulfonic acid is 1:1, 1:2 or 1:3;

In some embodiments, the pharmaceutically acceptable salt is L-malic acid salt, and the molar ratio of the compound represented by formula (I): L-malic acid is 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from phosphates, and the molar ratio of the compound represented by formula (I): phosphoric acid is 1:1, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from sulfates, and the molar ratio of the compound represented by formula (I): sulfuric acid is 1:1, 1:2 or 1:3;

In some embodiments, the pharmaceutically acceptable salt is selected from p-toluenesulfonic acid salts, and the molar ratio of the compound represented by formula (I): p-toluenesulfonic acid is 1:1, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from hydrochloric acid salts, and the molar ratio of the compound represented by formula (I): hydrochloric acid is 1:1, 1:2 or 1:3;

In some embodiments, the pharmaceutically acceptable salt is selected from maleic acid salts, and the molar ratio of the compound represented by formula (I): maleic acid is 1:1, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from hydrobromic acid salts, and the molar ratio of the compound represented by formula (I): hydrobromic acid is 1:1, 1:2 or 1:3;

In some embodiments, the pharmaceutically acceptable salt is selected from methanesulfonate salts, and the molar ratio of the compound represented by formula (I): methanesulfonic acid is 1:1, 1:2 or 1:3;

In some embodiments, the pharmaceutically acceptable salt is selected from citrate salts, and the molar ratio of the compound represented by formula (I): citric acid is 1:1, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from mandelic acid salts, and the molar ratio of the compound represented by formula (I): mandelic acid is 1:1, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from succinate, and the molar ratio of the compound represented by formula (I): succinic acid is 1:1, 1:2 or 1:3;

In some embodiments, the pharmaceutically acceptable salt is selected from fumarate salts, and the molar ratio of the compound represented by formula (I): fumaric acid is 1:0.5, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from hippurate salts, and the molar ratio of the compound represented by formula (I): hippuric acid is 1:1;

In some embodiments, the pharmaceutically acceptable salt is selected from salicylate, and the molar ratio of the compound represented by formula (I): salicylic acid is 1:1;

In some embodiments, the pharmaceutically acceptable salt is selected from 1,5-dinaphthylsulfonic acid salts, and the molar ratio of the compound represented by formula (I): 1,5-dinaphthylsulfonic acid is 1:1, 1:2;

In some embodiments, the pharmaceutically acceptable salt is selected from nicotinic acid salts, and the molar ratio of the compound represented by formula (I): nicotinic acid is 1:3;

In some embodiments, the above-mentioned pharmaceutically acceptable salt is amorphous.

The present invention relates to a maleate salt crystalline form 1 of a compound represented by formula (I); in some embodiments, using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 14.83°±0.2°, 16.53°±0.2°, 20.34°±0.2°, 22.87°±0.2°, 23.92°±0.2°; in some embodiments, using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 4.77°±0.2°, 6.75°±0.2°, 8.80°±0.2°, 14.83°±0.2°, 16.53°±0.2°, 20.34°±0.2°, 22.87°±0.2°, 23.92°±0.2°; 2.87°±0.2°, 23.92°±0.2°; in some embodiments, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 4.77°±0.2°, 6.75°±0.2°, 8.80°±0.2°, 10.97°±0.2°, 14.83°±0.2°, 16.53°±0.2°, 16.97°±0.2°, 18.68°±0.2°, 20.34°±0.2°, 21.08°±0.2°, 22.87°±0.2°, 23.92°±0.2°, 24.61°±0.2°; in some embodiments, Cu-Kα radiation is used, and its X-ray powder diffraction pattern is shown in Figure 1.

The present invention relates to a dimaleate crystal form 1 of a compound represented by formula (I); in some embodiments, using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 4.18°±0.2°, 8.24°±0.2°, 18.40°±0.2°, 20.48°±0.2°, 21.96°±0.2°; in some embodiments In the scheme, Cu-Kα radiation is used, and its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 4.18°±0.2°, 8.24°±0.2°, 18.40°±0.2°, 18.80°±0.2°, 20.48°±0.2°, 21.96°±0.2°, 23.66°±0.2°, 24.34°±0.2°; In some embodiments, Cu-Kα radiation is used, and its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 4.18°±0.2°, 8.24°±0.2°, 12.30±0.2°, 13.21°±0.2°, 16.34°±0.2°, 16.57°±0.2°, 18.40°±0.2°, 18.80°±0.2°, 19.70°±0.2°, 20. .2°, 20.00°±0.2°, 20.48°±0.2°, 20.81°±0.2°, 21.96°±0.2°, 23.66°±0.2°, 24.34°±0.2°, 25.73°±0.2°, 27.98°±0.2°; in some embodiments, Cu—Kα radiation is used, and its X-ray powder diffraction pattern is shown in Figure 6.

The present invention relates to a 2-naphthalenesulfonate salt crystal form 1 of a compound represented by formula (I); in some embodiments, using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 5.82°±0.2°, 17.52°±0.2°, 20.13°±0.2°, 21.16°±0.2°, 26.80°±0.2°; In some embodiments, Cu-Kα radiation is used, and its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 5.82°±0.2°, 12.58°±0.2°, 14.92°±0.2°, 17.52°±0.2°, 20.13°±0.2°, 21.16°±0.2°, 22.95°±0.2°, 26.80 °±0.2°; In some embodiments, Cu-Kα radiation is used, and its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 5.82°±0.2°, 7.10°±0.2°, 8.78°±0.2°, 11.59°±0.2°, 12.58°±0.2°, 14.92°±0.2°, 17.03°±0.2° , 17.52°±0.2°, 18.80°±0.2°, 20.13°±0.2°, 21.16°±0.2°, 21.72°±0.2°, 22.22°±0.2°, 22.95°±0.2°, 26.80°±0.2°; in some embodiments, Cu—Kα radiation is used, and its X-ray powder diffraction pattern is shown in Figure 11.

The present invention relates to an oxalate crystal form 1 of a compound represented by formula (I); in some embodiments, using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 19.69°±0.2°, 20.07°±0.2°, 23.75°±0.2°, 24.45°±0.2°, 26.82°±0.2°; in some embodiments, Using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 13.79°±0.2°, 17.86°±0.2°, 19.69°±0.2°, 20.07°±0.2°, 23.75°±0.2°, 24.45°±0.2°, 24.82°±0.2°, 26.82°±0.2°, 27.07° ±0.2°; in some embodiments, Cu-Kα radiation is used, and its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 6.91°±0.2°, 7.63°±0.2°, 13.52°±0.2°, 13.79°±0.2°, 17.86°±0.2°, 18.87°±0.2°, 19.69°±0.2°, 20.07 °±0.2°, 20.71°±0.2°, 23.75°±0.2°, 24.45°±0.2°, 24.82°±0.2°, 26.82°±0.2°, 27.07°±0.2°, 29.44°±0.2°, 31.28°±0.2°; in some embodiments, Cu—Kα radiation is used, and its X-ray powder diffraction pattern is shown in Figure 16.

The present invention relates to an amorphous dimethanesulfonate of a compound represented by formula (I), whose X-ray powder diffraction pattern is shown in FIG36 .

The present invention relates to a crystalline form 1 of a compound represented by formula (I); in some embodiments, using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 5.03°±0.2°, 15.35°±0.2°, 19.43°±0.2°, 19.88°±0.2°; in some embodiments, using Cu-Kα radiation, its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: The 2θ position has characteristic diffraction peaks: 5.03°±0.2°, 8.03°±0.2°, 13.25°±0.2°, 14.57°±0.2°, 14.88°±0.2°, 15.35°±0.2°, 19.43°±0.2°, 19.88°±0.2°, 23.83±0.2°, 24.71°±0.2°, 26.44°±0.2°, 29.93° ±0.2°; in some embodiments, Cu-Kα radiation is used, and its X-ray powder diffraction spectrum has characteristic diffraction peaks at the following 2θ positions: 5.03°±0.2°, 8.03°±0.2°, 8.99°±0.2°, 13.25°±0.2°, 14.57°±0.2°, 14.88°±0.2°, 15.35°±0.2°, 19.43°±0.2°, 19. 88°±0.2°, 20.17±0.2°, 22.32±0.2°, 22.55±0.2°, 23.83±0.2°, 24.71°±0.2°, 26.17±0.2°, 26.44°±0.2°, 29.49±0.2°, 29.93°±0.2°; in some embodiments, Cu-Kα radiation is used, and its X-ray powder diffraction pattern is shown in Figure 48.

The present invention relates to a method for preparing a pharmaceutically acceptable salt of a compound represented by formula (I), wherein the method comprises: a step of forming a salt with the compound represented by formula (I) and an acid; in some embodiments, the solvent used is selected from one or more of a C _1-6 halogenated alkane solvent, a C _2-6 ester solvent, a C _2-6 ether solvent, a C _1-6 alcohol solvent or water; in some embodiments, the solvent used is selected from one or more of dichloromethane, 1,2-dichloroethane, ethyl acetate, methanol, ethanol, isopropanol, propanol, ether, tetrahydrofuran and water.

The present invention relates to a pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of a crystalline or amorphous form of a pharmaceutically acceptable salt of any one of the compounds represented by formula (I) above, and a pharmaceutically acceptable carrier or excipient.

The present invention relates to the use of a crystalline or amorphous form of a pharmaceutically acceptable salt of any one of the compounds represented by formula (I) or the above-mentioned drug composition in the preparation of drugs for treating and inhibiting or degrading EGFR-related diseases (preferably cancer).

The present invention relates to the use of a crystalline or amorphous form of a pharmaceutically acceptable salt of any one of the compounds represented by formula (I) or the above-mentioned drug composition in the preparation of drugs for treating and inhibiting or degrading EGFR-related diseases (preferably cancer).

In some embodiments, the pharmaceutical composition of the present invention may be in the form of a unit dosage form (the amount of the main drug in a unit dosage form is also referred to as the "dosage strength").

The "effective amount" or "therapeutically effective amount" described in this application refers to the administration of a sufficient amount of the compound disclosed in this application, which will alleviate one or more symptoms of the disease or disorder (e.g., cancer) being treated to some extent. In some embodiments, the result is the reduction and/or alleviation of the signs, symptoms or causes of the disease, or any other desired changes in the biological system. For example, an "effective amount" for therapeutic use is the amount of the compound disclosed in this application required to provide a clinically significant reduction in disease symptoms. Examples of therapeutically effective amounts (calculated as free base) include, but are not limited to, 1-800 mg, 1-700 mg, 1-600 mg, 2-600 mg, 3-600 mg, 4-600 mg, 5-600 mg, 6-600 mg, 10-600 mg, 20-600 mg, 25-600 mg, 30-600 mg, 40-600 mg, 50-600 mg, 60-600 mg, 70-600 mg, 75-600 mg, 80-600 mg, 90-600 mg, 100-600 mg, 200-600 mg, 1-500 mg, 2-500 mg, 3-500 mg, 4-500 mg, 5-500 mg, 6-500 mg, 10-500 mg g, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500mg, 60-500mg, 70-500mg, 75-500mg, 80-500mg, 90-500mg, 100-500mg, 125-500mg, 150-500mg, 200-500mg, 250- 500mg, 300-500mg, 400-500mg, 5-400mg, 10-400mg, 20-400mg, 25-400mg, 30-400mg, 40-400mg, 50-400mg, 60-400mg, 70-400mg, 75-400mg, 80-400mg, 90 -400mg, 100-400mg, 125-400mg, 150-400mg, 200-400mg, 250-400mg, 300-400mg, 1-300mg, 2-300mg, 5-300mg, 10-300mg, 20-300mg, 25-300mg, 30-300mg, 40-300mg, 5 0-300mg, 60-300mg, 70-300mg, 75-300mg, 80-300mg, 90-300mg, 100-300mg, 125-300mg, 150-300mg, 200-300mg, 250-300mg, 1-200mg, 2-200mg, 5-200m g, 10-200mg, 20-200mg, 25-200mg, 30-200mg, 40-200mg, 50-200mg, 60-200mg, 70-200mg, 75-200mg, 80-200mg, 90-200mg, 100-200mg, 125-200mg, 150-200mg, 1-100 mg, 2-100mg, 5-100mg, 10-100mg, 15-100mg, 20-100mg, 25-100mg, 30-100mg, 40-100mg, 50-100mg, 60-100mg, 70-100mg, 75-100mg, 80-100mg, 90-100mg.

In some embodiments, examples of therapeutically effective amounts include, but are not limited to, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 300 mg.

A method for treating a disease in a mammal, the method comprising administering to a subject a therapeutically effective amount of a pharmaceutically acceptable salt or co-crystal of the compound of the present invention, preferably 1-800 mg, wherein the disease is preferably a disease related to the inhibition or degradation of EGFR (preferably cancer).

A method for treating a disease in a mammal, the method comprising administering a pharmaceutically acceptable salt or co-crystal of a compound of the present invention to a subject at a daily dose of 1-1000 mg/day, the daily dose may be a single dose or a divided dose. In some embodiments, the daily dose includes but is not limited to 10-1500 mg/day, 10-1000 mg/day, 10-800 mg/day, 25-800 mg/day, 50-800 mg/day, 100-800 mg/day, 200-800 mg/day, 25-400 mg/day, 50- In some embodiments, the daily dose includes but is not limited to 10 mg/day, 20 mg/day, 25 mg/day, 50 mg/day, 80 mg/day, 100 mg/day, 125 mg/day, 150 mg/day, 160 mg/day, 200 mg/day, 300 mg/day, 320 mg/day, 400 mg/day, 480 mg/day, 600 mg/day, 640 mg/day, 800 mg/day, 1000 mg/day.

The present invention relates to a kit which may include a composition in a single-dose or multi-dose form, wherein the kit contains a pharmaceutically acceptable salt or co-crystal of the compound of the present invention, and the amount of the pharmaceutically acceptable salt or co-crystal of the compound of the present invention is the same as that in the above-mentioned pharmaceutical composition.

The crystal form of the compound represented by formula (I) described in the present invention has excellent physical properties, including but not limited to solubility, dissolution rate, light resistance, low moisture absorption, high temperature resistance, and high humidity resistance. For example, the crystal form described in the present invention can significantly reduce the filtration time in the preparation process, shorten the production cycle, and save costs. The crystal form described in the present invention also has good light stability, thermal stability, and moisture stability, which can ensure the reliability of the crystal form during storage and transportation, thereby ensuring the safety of the preparation, and the crystal form does not need to be specially packaged to prevent the influence of light, temperature, and humidity, thereby reducing costs. The crystal form will not be degraded due to the influence of light, high temperature, and high humidity, thereby improving the safety of the preparation and the effectiveness after long-term storage. Patients taking the crystalline form do not have to worry about photosensitivity reactions to the formulation due to exposure to sunlight.

The crystalline form of the compound represented by formula (I) of the present invention degrades little or very little during storage or transportation at ambient temperature, has good thermal stability, can be stably maintained for a long time, and is suitable for standard preparation production processes.

The crystal form of the compound represented by formula (I) described in the present invention is suitable and convenient for mass preparation. The preparation prepared using the aforementioned crystal form can reduce irritation and improve absorption, so that the problem of metabolic rate can be solved, toxicity can be significantly reduced, safety can be improved, and the quality and efficacy of the preparation can be effectively guaranteed.

It can be understood that the expressions such as “preferably, ..., further having a characteristic diffraction peak at the following 2θ position” or “more preferably, ..., further having a characteristic diffraction peak at the following 2θ position” and the like in the present invention mean that on the basis of having a characteristic diffraction peak at the aforementioned 2θ position, there is further a characteristic diffraction peak at the aforementioned “following 2θ position”.

It is understood that the numerical values described and protected by the present invention are approximate values. Variations in the numerical values may be due to equipment calibration, equipment errors, crystal purity, crystal size, sample size and other factors.

The crystal structure of the present invention can be analyzed using various analytical techniques known to those of ordinary skill in the art, including but not limited to, X-ray powder diffraction (XRD), ion chromatography (IC), differential scanning calorimetry (DSC) and/or thermogravimetric analysis (TGA), also known as thermogravimetry (TG).

It is to be understood that the crystal form of the present invention is not limited to the characteristic spectra exactly the same as those described in the attached drawings disclosed in the present invention, such as XRD, DSC, TGA, and any crystal form having characteristic spectra that are substantially the same or essentially the same as those described in the attached drawings falls within the scope of the present invention.

It is understood that the melting peak height of a DSC curve depends on many factors related to sample preparation and instrument geometry, and the peak position is relatively insensitive to experimental details, as is well known in the art of differential scanning calorimetry (DSC). Therefore, in some embodiments, the crystalline compounds of the present invention are characterized by a DSC pattern with a characteristic peak position, having substantially the same properties as the DSC pattern provided in the accompanying figures of the present invention, with an error tolerance of ±3°C.

Unless otherwise specified, the technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention belongs. If there is a conflict, the definition provided in this application shall prevail. When a certain amount, concentration or other value or parameter is expressed in the form of a range, a preferred range, or a preferred upper numerical limit and a preferred lower numerical limit, it should be understood that it is equivalent to specifically revealing any range by combining any pair of upper range limits or preferred numerical values with any lower range limit or preferred numerical value, without considering whether the range is specifically disclosed. Unless otherwise specified, the numerical ranges listed herein are intended to include the endpoints of the range and all integers and fractions (decimals) within the range.

Unless otherwise stated, the terms used in the specification and claims have the following meanings.

The term "optional" or "optionally" as used herein means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur.

When used in conjunction with a numerical variable, "about" or "approximately" as used herein generally means that the value of the variable and all values of the variable are within the experimental error (e.g., within the 95% confidence interval for the mean) or within ±10% of the specified value, or within a wider range.

Unless otherwise specified, percentages, parts, etc. herein are all by weight.

"Amorphous" as used herein refers to any solid substance that is not ordered in three dimensions. In some cases, an amorphous solid can be characterized by known techniques, including XRPD crystal diffraction analysis, differential scanning calorimetry (DSC), solid-state nuclear magnetic resonance (ssNMR) spectroscopy analysis, or a combination of these techniques. As described below, an amorphous solid produces an XRPD pattern without obvious diffraction characteristic peaks.

As used herein, "crystalline form" or "crystal" refers to any solid material that exhibits a three-dimensional ordering, which, in contrast to an amorphous solid material, produces a characteristic XRPD pattern with well-defined peaks.

The "seed crystals" mentioned in the present invention refer to the formation of crystal nuclei by adding insoluble additives in the crystallization method to accelerate or promote the growth of enantiomer crystals with the same crystal form or stereo configuration.

The "pharmaceutical composition" mentioned in the present invention refers to a mixture of one or more compounds mentioned herein or their physiologically/pharmaceutically acceptable salts and other components, wherein the other components include physiologically/pharmaceutically acceptable carriers and excipients.

The "carrier" mentioned in the present invention refers to a carrier or diluent that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound.

The term "excipient" as used herein refers to an inert substance added to a pharmaceutical composition to further facilitate administration of the compound.

The "IC ₅₀ " mentioned in the present invention refers to the half-maximal inhibition concentration, which refers to the concentration when half of the maximum inhibition effect is achieved.

The "ether solvent" mentioned in the present invention refers to a chain compound or a ring compound containing an ether bond -O- and having 1 to 10 carbon atoms, and specific examples include but are not limited to: tetrahydrofuran, ethyl ether, propylene glycol methyl ether, methyl tert-butyl ether, isopropyl ether or 1,4-dioxane.

The "alcohol solvent" mentioned in the present invention refers to a group derived from one or more "hydroxyl" groups replacing one or more hydrogen atoms on a "C _{1 - 6} alkyl group". The "hydroxyl" and "C _{1 - 6} alkyl group" are as defined above, and specific examples include but are not limited to: methanol, ethanol, isopropanol, n-propanol, isopentanol or trifluoroethanol.

The "ester solvent" mentioned in the present invention refers to a combination of a lower organic acid containing 1 to 4 carbon atoms and a lower alcohol containing 1 to 6 carbon atoms, and specific examples include but are not limited to: ethyl acetate, isopropyl acetate or butyl acetate.

The "ketone solvent" mentioned in the present invention refers to a compound in which a carbonyl group (-C(O)-) is connected to two hydrocarbon groups. Ketones can be divided into aliphatic ketones, alicyclic ketones, aromatic ketones, saturated ketones and unsaturated ketones according to the different hydrocarbon groups in the molecule. Specific examples include but are not limited to: acetone, acetophenone, and 4-methyl-2-pentanone.

The "nitrile solvent" mentioned in the present invention refers to a group derived from one or more "cyano" groups replacing one or more hydrogen atoms on a "C _{1 - 6} alkyl group". The "cyano" and "C _{1 - 6} alkyl group" are as defined above, and specific examples include but are not limited to: acetonitrile or propionitrile.

The "halogenated hydrocarbon solvent" mentioned in the present invention refers to a group derived from one or more "halogen atoms" replacing one or more hydrogen atoms on a "C _{1 - 6} alkyl group". The "halogen atom" and "C _{1 - 6} alkyl group" are as defined above, and specific examples include but are not limited to: dichloromethane, 1,2-dichloroethane, chloroform or carbon tetrachloride.

The "crystal of the present invention", "crystal form of the present invention", "polymorph of the present invention" and the like described in the present invention can be used interchangeably.

The "room temperature" mentioned in the present invention generally refers to 4~30℃, preferably refers to 20±5℃.

The drying temperature of the present invention is generally 20-100°C, preferably 25-70°C, and can be either normal pressure drying or reduced pressure drying (vacuum drying). Preferably, the drying is performed under reduced pressure.

The "X-ray powder diffraction pattern (XRPD pattern)" mentioned in the present invention refers to the experimentally observed diffraction pattern or the parameters, data or values derived therefrom. The XRPD pattern is usually characterized by peak position (abscissa) and/or peak intensity (ordinate).

The "2θ or 2θ angle" described in the present invention refers to the diffraction angle, θ is the Bragg angle, which is based on the peak position expressed in degrees (°) set in the X-ray diffraction experiment, and is usually the horizontal coordinate unit in the diffraction spectrum. If the incident beam forms an angle θ with a certain lattice plane and the reflection is diffracted, the experimental setting needs to record the reflected beam at an angle of 2θ. It should be understood that the specific 2θ value of a specific crystal form mentioned in this article is intended to represent the 2θ value (expressed in degrees) measured using the X-ray diffraction experimental conditions described herein, and the error range of 2θ is ± 0.3, which can be ± 0.3, ± 0.2 or ± 0.1.

"Substantially the same" as used herein means that representative peak position and intensity variations are taken into account. For example, one skilled in the art will understand that peak position (2θ) will show some variation, typically as much as 0.1-0.2 degrees, and that the instrument used to measure diffraction will also cause some variation. In addition, one skilled in the art will understand that relative peak intensity will vary due to differences between instruments, as well as the degree of crystallinity, preferred orientation, prepared sample surface, and other factors known to one skilled in the art, and should be considered as only a qualitative measurement.

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

According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "9103 Guidance Principles for Hygroscopicity of Drugs" in Part IV of the 2020 edition of the Chinese Pharmacopoeia,

Deliquesce: absorb enough water to form a liquid;

Highly hygroscopic: weight gain after absorbing moisture is not less than 15%;

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

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

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

The crystal form disclosed in the present invention can be prepared by the following common methods for preparing crystal forms:

1. The volatility experiment is to allow the clear solution of the sample to evaporate openly at different temperatures until the solvent is dry.

2. The slurry experiment is to stir the supersaturated solution of the sample (with insoluble solids) at a certain temperature in different solvent systems.

3. The antisolvent experiment is to dissolve the sample in a good solvent, add an antisolvent, stir the precipitated solid for a short time and then filter it immediately.

4. The cooling crystallization experiment is to dissolve a certain amount of sample into the corresponding solvent at high temperature, and then stir and crystallize directly at room temperature or low temperature.

5. The polymer sample experiment is to add different types of polymer materials to the sample clear solution and leave it open at room temperature to evaporate until the solvent is dry.

6. The thermal method experiment is to treat the sample under certain thermal method crystallization conditions and cool it to room temperature.

7. The water vapor diffusion experiment is to place the sample in a certain humidity environment at room temperature.

The structures of the compounds were determined by nuclear magnetic resonance (NMR) or (and) mass spectrometry (MS). NMR shifts (δ) are given in units of 10 ^-6 (ppm). NMR measurements were performed using (Bruker Avance III 400 and Bruker Avance 300) NMR spectrometers, with deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD) as the solvent, and tetramethylsilane (TMS) as the internal standard.

MS was used for determination (Agilent 6120B (ESI) and Agilent 6120B (APCI)).

HPLC analysis was performed using an Agilent 1260DAD high pressure liquid chromatograph (Eclipse Plus C18, 150×4.6 mm).

The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from companies such as Titan Technology, Anage Chemical, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, and Bailingwei Technology.

The following detailed description of the implementation process and beneficial effects of the present invention is provided through specific embodiments, which is intended to help readers better understand the essence and features of the present invention, and is not intended to limit the scope of implementation of the present invention.

Example 1: Preparation of Compound 1

Step 1: Preparation of 1C

1A (16 g, 56.45 mmol) and 1B (14.8g, 59.27mmol) were dissolved in DMSO (200mL), potassium carbonate (23.5g, 0.17 mol) was added, and the mixture was stirred at 120℃ for 4 hours. The reaction solution was cooled to room temperature, and 300mL of water was added. A yellow solid was precipitated, which was filtered and washed with water three times. The filter cake was re-dissolved in dichloromethane, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (mobile phase: dichloromethane/methanol (V/V) = 50/1-15/1) to obtain 1C (22 g, yield: 76%).

Step 4: Preparation of 1E

1C (400 g, 0.78 mol) and 1D (158 g, 1.24 mol) were dissolved in a mixed solvent of 1,4-dioxane (1.6 L) and water (0.4 L). Potassium carbonate (216 g, 1.56 mol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (III) complex (38 g, 0.047 mol) were added. ), replaced with nitrogen 3 times, stirred at 90°C for overnight reaction, after the reaction was complete, reduced pressure and concentrated, then 500 mL of ethyl acetate was added to dilute the reaction solution, washed 3 times with water, washed once with saturated sodium chloride, the organic phase was dried over anhydrous sodium sulfate, reduced pressure and concentrated to obtain a crude 1E product, PE/EA=3/1 mixed solution (1600 mL) was added and slurried for 30 minutes, filtered, and the filter cake was reduced pressure and concentrated to obtain 1E (377 g, yield: 94%).

LCMS m/z =515.3 [M+H] ⁺

Step 5: Preparation of 1F

Dissolve 1E (377 g, 0.73 mol) in methanol (600 mL), add hydrogen chloride-dioxane solution (4 mol/L, 2.5 L), stir and react at room temperature for 60 min, reduce pressure and concentrate to obtain a crude product, add 2.5 L of water and 500 mL of concentrated hydrochloric acid to the crude product, extract with ethyl acetate (1.5 L*2), adjust the pH of the aqueous phase to 13 with sodium hydroxide solution, extract with dichloromethane (1.5 L*2), combine the organic phases and dry with anhydrous sodium sulfate, filter, reduce pressure and concentrate the filtrate, add PE/MTBE mixed solution (v/v=1/1, 1.2 L) to slurry, filter, reduce pressure and concentrate the filter cake to obtain 1F (256 g, yield: 85%).

Step 6: Preparation of 1G

1F (120 g, 0.29 mol) was dissolved in DMSO (600 mL), and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (80 g, 0.29 mmol) and diisopropylethylamine (112 g, 0.87 mmol) were added in sequence, and the mixture was reacted at 100 °C overnight. 2.0 L of water was added, and the solid was precipitated and filtered. The filter cake was dissolved and extracted with 200 mL of dichloromethane. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (mobile phase: dichloromethane/methanol (V/V) = 100/1-10/1) to obtain 1G (186 g, yield: 96%).

Step 7: 1H Preparation

Dissolve 1G (130 g, 0.19 mmol) in a mixture of THF (1.3 L) and water (400 mL), add ammonium chloride (51 g, 0.95 mmol) and zinc powder (62 g, 0.95 mmol), slowly raise the temperature to 40-60°C and react for 1-2 hours. Cool the reaction solution to room temperature, filter, wash the filter cake with 1L of DCM, combine the organic phases, wash the organic phases with 0.5L ammonia water and 0.5L saturated brine in sequence, dry over anhydrous sodium sulfate and concentrate under reduced pressure to obtain 1H as a yellow solid.

LCMS m/z = 641.3 [M+H] ⁺

Step 8: Preparation of Compound 1

1H (80 g, 126 mmol) and 1M (51 g, 126 mmol) were dissolved in DMF (500 mL), p-toluenesulfonic acid monohydrate (48 g, 252 mmol) was added, and the mixture was reacted at 100 °C overnight. After cooling to room temperature, 800 mL of saturated sodium bicarbonate aqueous solution was slowly added, and the mixture was filtered. The filter cake was dissolved and extracted with dichloromethane. The organic phase was dried with anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol (V/V) = 100/1-100/4). Column chromatography gave 64 g of compound 1 (64 g, yield: 50%).

LCMS m/z =502.7 [(M+2H)/2] ⁺ .

¹ H NMR (400 MHz, D ₂ O/CF ₃ COOD(v/v=1:1)) δ 8.23 (s, 1H), 7.96 – 7.75 (m, 5H), 7.46 (s, 1H), 7.35 ( s, 1H), 7.31 – 7.15 (m, 2H), 6.91 (d, 1H), 5.12 (dd, 1H), 4.22 – 4.07 (m, 5H), 4.03 (s, 3H), 3.93 – 3.70 (m, 6H), 3.62-3.48 (m, 2H), 3.39 – 3.18 (m, 4H), 2.95 – 2.85 (m, 2H), 2.82 – 2.67 (m, 1H), 2.63 – 2.44 (m, 1H), 2.37 – 2.16 (m, 3H), 2.12 – 1.88 (m, 9H), 1.19 – 1.08 (m, 2H), 0.73 – 0.65 (m, 2H).

Example 2: Preparation of Maleate Form 1 of Compound 1

Take about 100 mg of compound 1, add 2.0 ml of tetrahydrofuran, and add 0.2 ml of tetrahydrofuran solution mixed with 13 mg of maleic acid. Stir overnight at room temperature, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain maleate salt form 1 of compound 1, whose XRD, DSC, TGA, isothermal adsorption curve and DVS are shown in Figures 1-5 respectively.

Example 3: Preparation of dimaleic acid form 1 of compound 1

Take about 100 mg of compound 1, add 2.0 ml of tetrahydrofuran, add 0.4 ml of tetrahydrofuran solution mixed with 26 mg of maleic acid, and precipitate. Stir at room temperature for 2 days, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the dimaleate crystal form 1 of compound 1 , whose XRD, DSC, TGA, isothermal adsorption curve and DVS are shown in Figures 6-10 respectively.

Through 1H NMR (400 MHz, DMSO-d ₆ ) peak shift analysis, the chemical shift 5.12 (dd, J = 12.9, 5.4 Hz, 1H) of compound 1 is the -CH peak at position 38, and the peak at 6.15 (s, 4H) is the -CH peak of maleic acid. The ratio is 1:4, so it can be analyzed that the ratio of compound 1 to maleic acid is 1:2.

Example 4: Preparation of 2-naphthalenesulfonate salt of compound 1, Form 1

Take about 100 mg of compound 1, add 2.0 ml of chloroform, and add 0.2 ml of ethanol solution mixed with 23 mg of 2-naphthalenesulfonic acid. Stir at room temperature for 2 days, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain 2- naphthalenesulfonic acid salt form 1 of compound 1. Its XRD, DSC, TGA, isothermal adsorption curve and DVS are shown in Figures 11-15 respectively.

Through 1H NMR (400 MHz, DMSO-d ₆ ) peak shift analysis, the chemical shift 5.12 (dd, J = 12.9, 5.4 Hz, 1H) of compound 1 is the -CH peak at position 38, and the peak at 8.31 – 8.14 (m, 4H) is the peak of 2-naphthalenesulfonic acid. The ratio is 1:4, so it can be analyzed that the ratio of compound 1 to 2-naphthalenesulfonic acid is 1:1.

Example 5: Preparation of Oxalate Form 1 of Compound 1

Take about 50 mg of compound 1, add 2.0 ml of dichloromethane, and add 0.2 ml of ethanol solution mixed with 15 mg of oxalic acid. After mixing at room temperature, stir at 4°C overnight, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain oxalate salt form 1 of compound 1 , whose XRD, DSC, and TGA are shown in Figures 16-18 respectively.

Ion chromatography (IC) detection results showed that the salt ratio was 1:1. Peak area (μS*min) Oxalate ion concentration (mg/mL) Weight of oxalate ions in salt (mg) Oxalate ion content in salt (%) Salt ratio 2.427 0.0868445 0.87 8.51 1.06

Example 6: Preparation of amorphous benzenesulfonate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.1 ml of methanol solution mixed with 10 mg of benzenesulfonic acid. Stir overnight at room temperature, add 2.0 mL of n-heptane and 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the solid at room temperature overnight to obtain the amorphous benzenesulfonate salt of compound 1. Its XRD is shown in Figure 19.

Example 7: Preparation of amorphous tribenzenesulfonate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of tetrahydrofuran, and add 0.3 ml of tetrahydrofuran solution mixed with 29 mg of benzenesulfonic acid. Stir overnight at room temperature to precipitate solid, centrifuge, and vacuum dry the solid at room temperature overnight to obtain the tribenzenesulfonate salt of compound 1 , whose XRD is shown in Figure 20.

Example 8: Preparation of amorphous di-L-apple acid salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.2 ml of methanol solution mixed with 15 mg of L-apple acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous di-L-apple acid salt of compound 1. Its XRD is shown in Figure 21.

Example 9: Preparation of amorphous phosphate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.1 ml of ethanol solution mixed with 6 mg of phosphoric acid, and precipitate solid. Stir overnight at room temperature, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous phosphate of compound 1, whose XRD is shown in Figure 22.

Example 10: Preparation of amorphous diphosphate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 13 mg of phosphoric acid, and precipitate solid. Stir overnight at room temperature, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous diphosphate of compound 1, whose XRD is shown in Figure 23.

Example 11: Preparation of amorphous sulfate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.1 ml of methanol solution mixed with 5.5 mg of sulfuric acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the solid at room temperature overnight to obtain the amorphous sulfate salt of compound 1. Its XRD is shown in Figure 24.

Example 12: Preparation of the amorphous disulfate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.2 ml of methanol solution mixed with 11 mg of sulfuric acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the disulfate salt of compound 1 in amorphous form, whose XRD is shown in Figure 25.

Example 13: Preparation of amorphous trisulfate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of tetrahydrofuran, and add 0.3 ml of tetrahydrofuran solution mixed with 16.5 mg of sulfuric acid. Stir overnight at room temperature to precipitate solid, centrifuge, and vacuum dry the solid at room temperature overnight to obtain the trisulfate salt of compound 1 in amorphous form, whose XRD is shown in Figure 26.

Example 14: Preparation of amorphous di-p-toluenesulfonate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, and add 0.2 ml of methanol solution mixed with 19 mg of p-toluenesulfonic acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, precipitate solid, stir for 6 hours, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous di-p-toluenesulfonate salt of compound 1, whose XRD is shown in Figure 27.

Example 15: Preparation of amorphous hydrochloride of compound 1

Take about 50 mg of the compound represented by formula (I), add 1.0 ml of dichloromethane, and add 0.1 ml of a methanol solution mixed with 6 mg of hydrochloric acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, precipitate solid, stir for 6 hours, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous hydrochloride of compound 1, whose XRD is shown in Figure 28.

Example 16: Preparation of the amorphous dihydrochloride salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.2 ml of methanol solution mixed with 11 mg of hydrochloric acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the dihydrochloride amorphous form of compound 1, whose XRD is shown in Figure 29.

Example 17: Preparation of the amorphous trihydrochloride salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of tetrahydrofuran, add 0.3 ml of tetrahydrofuran solution mixed with 17 mg of hydrochloric acid, and precipitate solid. Stir overnight at room temperature, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the trihydrochloride amorphous form of compound 1, whose XRD is shown in Figure 30.

Example 18: Preparation of amorphous di-2-naphthalenesulfonate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, and add 0.2 ml of ethanol solution mixed with 23 mg of 2-naphthalenesulfonic acid. Stir overnight at room temperature, add 2.0 mL of n-heptane, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous di -2- naphthalenesulfonic acid salt of compound 1 , whose XRD is shown in Figure 31.

Example 19: Preparation of the amorphous hydrobromide salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.1 ml of methanol solution mixed with 11 mg of hydrobromic acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous hydrobromide of compound 1 , whose XRD is shown in Figure 32.

Example 20: Preparation of amorphous dihydrobromide salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.2 ml of methanol solution mixed with 22 mg of hydrobromic acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous dihydrobromide salt of compound 1 , whose XRD is shown in Figure 33.

Example 21: Preparation of amorphous trihydrobromide salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of tetrahydrofuran, add 0.3 ml of tetrahydrofuran solution mixed with 33 mg of hydrobromic acid, and precipitate solid. Stir overnight at room temperature, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous trihydrobromide salt of compound 1 , whose XRD is shown in Figure 34.

Example 22: Preparation of amorphous methanesulfonate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, and add 0.1 ml of methanol solution mixed with 5 mg of methanesulfonic acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous methanesulfonate of compound 1 , whose XRD is shown in Figure 35.

Example 23: Preparation of the amorphous dimethanesulfonate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of dichloromethane, add 0.2 ml of methanol solution mixed with 11 mg of methanesulfonic acid. Stir overnight at room temperature, add 2.0 mL of methyl tert-butyl ether, stir for 6 hours, precipitate solid, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous dimethanesulfonate of compound 1 , whose XRD is shown in Figure 36.

Through 1H NMR (400 MHz, DMSO-d6) peak shift analysis, the chemical shift 5.12 (dd, J = 12.9, 5.4 Hz, 1H) of compound 1 is the -CH peak at position 38, and the peak at 2.70 (s, 6H) is the peak of methanesulfonic acid. The ratio is 1:6, so it can be analyzed that the ratio of compound 1 to methanesulfonic acid is 1:2.

Example 24: Preparation of the amorphous tris(methysulfonate) salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of tetrahydrofuran, add 0.3 ml of tetrahydrofuran solution mixed with 16 mg of methanesulfonic acid, and precipitate solid. Stir overnight at room temperature, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous tris-methanesulfonate of compound 1 , whose XRD is shown in Figure 37.

Example 25: Preparation of amorphous mandelate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 9 mg of mandelic acid, stir the solution at 4°C overnight, no solid precipitates, add about 4 mL of n-heptane to obtain a suspension, stir at 4°C overnight. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous mandelic acid salt of compound 1 , whose XRD is shown in Figure 38.

Example 26: Preparation of the amorphous dimandelate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 18 mg of mandelic acid, stir the solution at 4°C overnight, no solid precipitates, add about 4 mL of n-heptane to obtain a suspension, stir at 4°C overnight. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous dimandelate salt of compound 1 , whose XRD is shown in Figure 39.

Example 27: Preparation of amorphous succinate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 7 mg of succinic acid, stir the solution at 4°C overnight, no solid precipitates, add about 4 mL of n-heptane to obtain a suspension, stir at 4°C overnight. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous succinate salt of compound 1 , whose XRD is shown in Figure 40.

Example 28: Preparation of amorphous salicylate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 8 mg of salicylic acid, stir the solution at 4°C overnight, no solid precipitates, add about 4 mL of n-heptane to obtain a suspension, stir at 4°C overnight. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous succinate salt of compound 1 , whose XRD and DSC are shown in Figure 41.

Example 29: Preparation of amorphous 1,5-naphthalene disulfonic acid salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 22 mg of 1,5-naphthalene disulfonic acid, stir the solution at 4°C overnight, and precipitate solid. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous 1,5- naphthalene disulfonic acid salt of compound 1 , whose XRD is shown in Figure 42.

Example 30: Preparation of amorphous di-1,5-naphthalene disulfonic acid salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 44 mg of 1,5-naphthalene disulfonic acid, stir the solution at 4°C overnight, and precipitate solid. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous di -1,5- naphthalene disulfonic acid salt of compound 1 , whose XRD is shown in Figure 43.

Example 31: Preparation of amorphous hemifumarate of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 7 mg of fumaric acid, stir the solution at 4°C overnight, and precipitate solid. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous hemifumarate of compound 1 , whose XRD is shown in Figure 44.

Example 32: Preparation of amorphous difumarate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 14 mg of fumaric acid, stir the solution at 4°C overnight, precipitate solid, add about 4 mL of n-heptane to obtain a suspension, stir at 4°C overnight. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous difumarate salt of compound 1 , whose XRD is shown in Figure 45.

Example 33: Preparation of amorphous trinicotinate salt of compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 8 mg of nicotinic acid, stir the solution at 4°C overnight, no solid is precipitated, add about 4 mL of n-heptane to obtain a suspension, stir at 4°C overnight. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous trinicotinic acid salt of compound 1 , whose XRD is shown in Figure 46.

Example 34: Preparation of the Amorphous Hippurate Salt of Compound 1

Take about 50 mg of compound 1, add 1.0 ml of chloroform, add 0.2 ml of ethanol solution mixed with 11 mg of hippuric acid, stir the solution at 4°C overnight, no solid precipitates, add about 4 mL of n-heptane to obtain a suspension, stir at 4°C overnight. Centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain the amorphous hippurate salt of compound 1 , whose XRD is shown in Figure 47.

Example 35: Preparation of Form 1 of Compound 1

Take about 50 mg of compound 1, add 1.0 ml of tetrahydrofuran, stir the solution at room temperature overnight, centrifuge, and vacuum dry the obtained solid at room temperature overnight to obtain Form 1 of compound 1 , whose XRD, TGA, and DSC are shown in Figures 48-50.

X- X-ray powder diffraction instrument (XRD)/DSC/TGA/DVS/IC Test

XRD/DSC/TGA/DVS test parameters are shown in Table 1.

Table 1 XRD/DSC/TGA/DVS test instruments and parameters X- ray Powder Diffraction (XRPD) and Hot Stage XRPD Instruments Model Bruker D8 Advance Diffractometer No. LY-01-034 Technical indicators Copper target with 1.54Å Kα radiation (40 KV, 40 mA), θ-2θ goniometer, nickel filter, Lynxeye detector Collection software Diffrac Plus XRD Commander Calibration material Corundum (Al ₂ O ₃ ) Analysis software MDI Jade appendix Non-reflective sample plate Specifications 24.6 mm diameter x1.0 mm thickness Manufacturer MTI Corporation Variable temperature hot plate Manufacturer Shanghai Micro-Image Instrument Technology Development Co., Ltd. Sample sheet quality Copper Plate Parameters Detection Angle 3°-40° 2θ/3°-30° 2θ (hot stage XRPD) Step length 0.02° 2θ speed 0.2 s.step ^-1 Sample volume ＞2 mg Remarks Unless otherwise specified, samples were not ground before testing. Differential Scanner (DSC) Instruments Model TA Instruments Q200 DSC No. LY-01-002 Control software Thermal Advantage Analysis software Universal Analysis Sample Plate Aluminum crucible (with lid but no holes) Parameters Sample volume 0.5 mg-5 mg Protective gas Nitrogen Gas flow rate 50 mL/min Testing methods Equilibrate at 0℃; Ramp 10℃/min to 200℃~280℃ Thermogravimetric Analyzer (TGA) Instruments Model TA Instruments Q500 TGA No. LY-01-003 Control software Thermal Advantage Analysis software Universal Analysis Sample Plate Platinum Crucible Parameters Sample quantity 1 mg-10 mg Protective gas Nitrogen Gas flow rate 40 mL/min Testing methods Hi-Res sensitivity 3.0; Ramp 10.00℃/min, res 5.0 to 120.00℃/150.00℃; Ramp 10.00℃/min to 350℃ Dynamic VIS (Dynamic Visual Sensor ) Instruments Model Intrinsic PLUS No. LY-01-165 Control software DVS Control Analysis software Microsoft Excel Sample Plate Stainless steel plate Parameters Sample quantity 1 mg-10 mg Protective gas Nitrogen Gas flow rate 200sccm Test method 1 Equilibrium temperature: 25℃; Initial humidity: 60%; Equilibrium time: 90 min; dm/dt (%): < 0.0100 for 15.00 min; Humidity gradient: 10% every 90 min to 80%; dm/dt (%): < 0.0100 for 15.00 min; Humidity gradient: 10% every 90 min to 0%; dm/dt (%): < 0.0100 for 15.00 min; Humidity gradient: 10% every 90 min to 60% Detection method 2 Equilibrium temperature: 25℃; Initial humidity: 0%; Equilibrium time: 90 min; dm/dt (%): ＜ 0.0100 for 15.00 min; Humidity gradient: 10% every 90 min to 80%; dm/dt (%): (%) ＜ 0.0100 for 15.00 min; Humidity gradient: 10% every 90 min to 0%; Judgment criteria No or almost no hygroscopicity ＜ 0.2% Slightly hygroscopic ≥ 0.2%, < 2% Hygroscopic ≥ 2%, < 15% Very hygroscopic ≥ 15% deliquescence Aqueous solution

Ion Chromatography (IC) Instrument Information and Method Parameters Device Name Ion Chromatography (Anion) Equipment Model Dionex ICS-900 IC Equipment Number LY-01-012 Chromatographic column Dionex, IonPac ^TM AS11, 4X250 mm Protective column Dionex, IonPac ^TM AG11, 4X50 mm Suppressor Dionex, ASRS®300 4-mm Method parameters Suppressor current 50 mA Flow rate 1.0 mL/min (isocratic) Injection volume 10 μL Background conductivity 2.520 μS Background Pressure 1547 psi Detection time 10 min Eluent 20mM Sodium Hydroxide Eluent 3. Testing standards: General rules of the four parts of the 2015 edition of the Chinese Pharmacopoeia 4. Testing Process 1) Preparation of eluent: Weigh 1.6 g of solid sodium hydroxide, dilute to 2 L with purified water, sonicate for 10 min, and remove bubbles. 2) Preparation of standard solution: L1: Take 50 mg of solid oxalic acid dihydrate, dissolve it in purified water to 50 mL. L2: Use a pipette to take 10 mL of "L1" solution, dissolve it in purified water to 20 mL. L3: Use a pipette to take 5 mL of "L1" solution, dissolve it in purified water to 20 mL. L4: Use a pipette to take 2 mL of "L1" solution, dissolve it in purified water to 20 mL. L5: Use a pipette to take 1 mL of "L1" solution, dissolve it in purified water to 20 mL. 3) Preparation of sample solution: Weigh about 10 mg of sample, add 20 mM sodium hydroxide to make up to 10 mL. If the sample is insoluble, ultrasonicate at room temperature for about 30 minutes, filter, and the filtrate is the sample solution to be tested after filtering through a 0.45 μm aqueous filter membrane and then injecting.

Table 2: XRD peak list of maleate salt of compound 1, form 1 2-Theta d Height I% Area I% 4.769 18.514 136 14.2 1641 10.2 6.749 13.0865 129 13.4 1677 10.4 8.795 10.0462 136 14.2 1505 9.3 10.168 8.6926 52 5.4 402 2.5 10.971 8.0579 80 8.3 1070 6.6 11.505 7.6848 50 5.2 1027 6.4 11.803 7.4919 65 6.8 1022 6.3 13.061 6.7728 64 6.7 1110 6.9 14.83 5.9685 167 17.4 3233 20.1 16.528 5.3591 295 30.7 5433 33.7 16.969 5.2206 120 12.5 1969 12.2 17.842 4.9672 62 6.5 459 2.8 18.683 4.7454 120 12.5 2166 13.4 20.341 4.3622 960 100 16120 100 21.078 4.2113 99 10.3 1164 7.2 21.795 4.0743 54 5.6 314 1.9 22.871 3.885 146 15.2 2829 17.5 23.922 3.7168 201 20.9 6454 40 24.608 3.6147 84 8.8 638 4 25.756 3.4561 61 6.4 753 4.7 27.216 3.2739 51 5.3 1699 10.5 27.697 3.2182 63 6.6 1489 9.2 28.667 3.1114 51 5.3 944 5.9 29.043 3.072 46 4.8 1258 7.8 30.167 2.96 42 4.4 831 5.2 32.347 2.7654 40 4.2 591 3.7 33.161 2.6993 42 4.4 303 1.9

Table 3: XRD peak list of dimaleate salt of compound 1, form 1 2-Theta d Height I% Area I% 4.182 21.1097 1285 51.3 12211 49.7 6.838 12.9168 114 4.6 1297 5.3 8.239 10.7225 720 28.8 7825 31.8 8.753 10.0945 87 3.5 1356 5.5 9.725 9.0877 116 4.6 949 3.9 10.429 8.4752 124 5 996 4.1 11.228 7.8738 64 2.6 784 3.2 12.300 7.1898 303 12.1 2945 12 12.916 6.8487 95 3.8 1310 5.3 13.213 6.6951 216 8.6 3784 15.4 13.581 6.5146 98 3.9 1745 7.1 15.076 5.8718 180 7.2 2273 9.2 16.378 5.408 348 13.9 4320 17.6 16.569 5.3458 367 14.7 5034 20.5 17.403 5.0914 195 7.8 1355 5.5 18.399 4.818 2504 100 24582 100 18.801 4.7159 503 20.1 9937 40.4 19.998 4.4362 197 7.9 1210 4.9 20.475 4.3339 762 30.4 16689 67.9 20.814 4.2642 278 11.1 7362 29.9 21.032 4.2206 97 3.9 1428 5.8 21.960 4.0442 702 28 6957 28.3 23.102 3.8468 153 6.1 1394 5.7 23.659 3.7575 675 27 6909 28.1 24.343 3.6534 563 22.5 10533 42.8 24.605 3.6151 152 6.1 3272 13.3 24.954 3.5653 94 3.8 653 2.7 25.733 3.4592 306 12.2 3876 15.8 27.503 3.2404 122 4.9 1698 6.9 27.979 3.1863 285 11.4 3619 14.7 28.767 3.1008 97 3.9 2149 8.7 29.011 3.0753 138 5.5 3171 12.9 30.231 2.9539 118 4.7 1114 4.5 30.797 2.9009 86 3.4 959 3.9 30.999 2.8825 54 2.2 323 1.3 31.963 2.7977 144 5.8 1815 7.4 33.72 2.6558 48 1.9 1107 4.5 33.99 2.6354 41 1.6 1097 4.5 34.208 2.6191 46 1.8 1272 5.2 35.037 2.559 42 1.7 177 0.7 36.231 2.4773 67 2.7 399 1.6 37.338 2.4064 47 1.9 568 2.3

Table 4: XRD peak list of 2-naphthalenesulfonate salt of compound 1, form 1 2-Theta d Height I% Area I% 5.823 15.1651 330 43.3 4789 38.3 7.096 12.4478 84 11 809 6.5 8.775 10.0689 79 10.4 1324 10.6 10.310 8.5732 47 6.2 814 6.5 11.591 7.6283 108 14.2 1870 14.9 12.583 7.029 165 21.6 2897 23.2 14.337 6.1726 55 7.2 634 5.1 14.924 5.9311 166 21.8 3299 26.4 15.886 5.5741 52 6.8 812 6.5 17.026 5.2035 91 11.9 2477 19.8 17.520 5.0577 254 33.3 5187 41.5 18.797 4.7171 86 11.3 1842 14.7 20.130 4.4075 186 24.4 2149 17.2 21.159 4.1954 763 100 12512 100 21.719 4.0886 86 11.3 2767 22.1 22.220 3.9974 121 15.9 1076 8.6 22.951 3.8718 154 20.2 3505 28 25.355 3.5099 47 6.2 1256 10 25.671 3.4674 49 6.4 993 7.9 26.798 3.324 169 22.1 4650 37.2 29.030 3.0733 53 6.9 963 7.7 29.868 2.989 65 8.5 1229 9.8 32.590 2.7453 40 5.2 874 7 35.063 2.5571 38 5 564 4.5

Table 5: XRD peak list of oxalate salt form 1 of compound 1 2-Theta d Height I% Area I% 5.325 16.5810 51 14.5 999 11.2 6.907 12.7868 168 47.7 2877 32.4 7.634 11.5712 82 23.3 1236 13.9 11.800 7.4938 38 10.8 469 5.3 13.517 6.5453 129 36.6 2224 25.0 13.786 6.4182 185 52.6 3378 38.0 14.642 6.0449 38 10.8 463 5.2 15.885 5.5745 52 14.8 547 6.2 17.862 4.9617 268 76.1 3772 42.5 18.874 4.6979 109 31.0 1063 12.0 19.694 4.5040 276 78.4 4990 56.2 20.072 4.4202 352 100.0 5965 67.1 20.707 4.2861 143 40.6 1768 19.9 22.688 3.9160 67 19.0 684 7.7 23.751 3.7431 352 100.0 4042 45.5 24.454 3.6371 311 88.4 7781 87.6 24.819 3.5844 211 59.9 4313 48.5 26.174 3.4018 69 19.6 719 8.1 26.821 3.3212 351 99.7 8885 100.0 27.065 3.2918 196 55.7 3541 39.9 27.620 3.2270 34 9.7 209 2.4 29.441 3.0314 91 25.9 1945 21.9 30.478 2.9306 45 12.8 415 4.7 31.277 2.8575 120 34.1 2560 28.8 32.894 2.7206 35 9.9 245 2.8 33.753 2.6533 63 17.9 1057 11.9

Table 6: XRD peak list of Form 1 of Compound 1 2-Theta d Height I% Area I% 5.03 17.5397 863 38.5 6177 25.2 7.28 12.1363 68 3.0 609 2.5 8.03 11.0008 514 22.9 4264 17.4 8.70 10.1592 55 2.5 484 2.0 8.99 9.8236 265 11.8 1978 8.1 10.54 8.3854 55 2.5 262 1.1 10.90 8.1137 87 3.9 695 2.8 11.21 7.8871 83 3.7 1254 5.1 11.63 7.6024 86 3.8 1435 5.9 12.07 7.3241 65 2.9 783 3.2 12.98 6.8146 259 11.5 1756 7.2 13.25 6.6746 244 10.9 2665 10.9 14.57 6.0763 428 19.1 3261 13.3 14.88 5.9501 318 14.2 2747 11.2 15.35 5.7680 851 37.9 7059 28.8 15.94 5.5552 92 4.1 775 3.2 16.55 5.3509 106 4.7 966 3.9 16.88 5.2486 53 2.4 677 2.8 17.92 4.9451 275 12.3 1591 6.5 18.26 4.8540 117 5.2 946 3.9 18.81 4.7134 153 6.8 1535 6.3 19.06 4.6522 233 10.4 1584 6.5 19.43 4.5656 2244 100.0 24515 100.0 19.88 4.4618 974 43.4 12475 50.9 20.17 4.3999 126 5.6 2046 8.3 20.63 4.3009 93 4.1 1057 4.3 20.84 4.2591 146 6.5 1483 6.0 21.64 4.1031 51 2.3 453 1.8 22.32 3.9792 122 5.4 2313 9.4 22.55 3.9392 103 4.6 2278 9.3 22.80 3.8974 81 3.6 916 3.7 23.09 3.8492 57 2.5 461 1.9 23.41 3.7976 170 7.6 1516 6.2 23.83 3.7306 923 41.1 8093 33.0 24.42 3.6419 102 4.5 1926 7.9 24.71 3.6005 229 10.2 2524 10.3 25.29 3.5186 106 4.7 1432 5.8 26.17 3.4022 133 5.9 2100 8.6 26.44 3.3683 142 6.3 2943 12.0 27.01 3.2990 41 1.8 250 1.0 27.60 3.2290 64 2.9 899 3.7 27.96 3.1880 47 2.1 484 2.0 28.48 3.1319 46 2.0 229 0.9 28.82 3.0953 63 2.8 1018 4.2 29.25 3.0505 141 6.3 1978 8.1 29.49 3.0266 144 6.4 2263 9.2 29.93 2.9833 215 9.6 2976 12.1 30.17 2.9600 120 5.3 1871 7.6 30.86 2.8950 56 2.5 1426 5.8 31.30 2.8558 55 2.5 1423 5.8 32.04 2.7915 46 2.0 771 3.1 32.93 2.7176 41 1.8 696 2.8 33.16 2.6990 38 1.7 700 2.9 33.72 2.6555 34 1.5 912 3.7 34.06 2.6299 46 2.0 900 3.7 34.52 2.5958 36 1.6 290 1.2 34.74 2.5798 38 1.7 212 0.9 37.96 2.3685 40 1.8 656 2.7 38.95 2.3103 35 1.6 598 2.4 39.34 2.2886 49 2.2 1097 4.5

Table 7. Solubility of the pharmaceutically acceptable salt of compound 1 in amorphous water (mg/mL) Salt type Solubility in water (mg/mL) Diphosphate amorphous 7.66 Dihydrochloride amorphous 6.04 Succinate amorphous 6.73 Dimesylate amorphous 9.66

Table 8. Hygroscopicity of the crystalline form of the pharmaceutically acceptable salt of Compound 1 Salt type Characteristic crystal form Hygroscopicity ( 0%RH to 80%RH ) Maleate Crystal form 1 1.8% (w/w), slightly hygroscopic, crystal form unchanged Dimaleate Crystal form 1 1.9% (w/w), slightly hygroscopic, crystal form unchanged 2-Naphthalenesulfonate Crystal form 1 1.8% (w/w), slightly hygroscopic, crystal form unchanged

Biological activity test

Test Example 1: Proliferation Inhibitory Activity of NCI-H1975 (EGFR-L858R-T790M) and A431 (EGFR-WT) Cells

NCI-H1975 (EGFR-L858R-T790M) and A431 (EGFR-WT) cells were purchased from ATCC and cultured in RPMI1640+10% FBS and DMEM+10% FBS, respectively, at 37 ºC in a 5% CO ₂ incubator. On the first day, NCI-H1975 (EGFR-L858R-T790M) and A431 (EGFR-WT) cells in the exponential growth phase were collected and live cells were counted using an automatic cell analyzer (countstar). The cell suspension was adjusted with culture medium and plated on 96-well cell culture plates. 1000 NCI-H1975 (EGFR-L858R-T790M) cells and 3000 A431 cells were plated on each well. On the second day, the culture medium was removed and 90 µL of fresh culture medium and 10 µL of different concentrations of compounds were added to each well. The final DMSO concentration in each well was 0.1%. The plates were cultured in a 37 ºC, 5% CO ₂ incubator for 72 hours. After 72 hours of drug treatment, 50 μL of pre-melted and room temperature CTG solution (Promega, G7572) was added to each well and mixed with a microplate shaker for 2 minutes. After standing at room temperature for 10 minutes, the fluorescence signal value was measured with an enzyme labeler (PHERAstar FSX).

Cell survival rate was calculated using the formula V _sample /V _{vehicle control} x100%, where V _sample is the reading of the drug-treated group and V _{vehicle control} is the average value of the solvent control group. Origin9.2 software was used to draw the S-shaped dose-survival rate curve using a nonlinear regression model and calculate the IC ₅₀ value.

Table 9 Proliferation inhibition results of NCI-H1975 (EGFR-L858R-T790M) and A431 (EGFR-WT) cells Compound No. IC50(μM) NCI-H1975(EGFR-L858R-T790M) IC50(μM) A431(EGFR-WT) Compound 1 0.054 ≥10

Conclusion: The compound has good proliferation inhibitory activity against NCI-H1975 (EGFR-L858R-T790M) cells, but poor proliferation inhibitory activity against A431 (EGFR-WT) cells, and has good selectivity.

Test Example 2: Proliferation Inhibitory Activity on Cells NCI-H1975 EGFR-L858R-T790M-C797S

Cells NCI-H1975 EGFR-L858R-T790M-C797S were cultured in a 37 °C, 5% CO ₂ incubator, and the culture medium was RPMI1640+10%FBS+100 µg/mL hygromycin. Cells in the exponential growth phase were collected, and the cell suspension was adjusted to an appropriate concentration with a culture medium without hygromycin and then plated on a 96-well plate, with a plate density of 1500 cells/well and a volume of 90 µL. 10 µL of compounds of different concentrations were added, and a solvent control group of cells plus DMSO was set up, and the concentration of DMSO was 0.1%. The cell culture plate was placed in a 37 °C, 5% CO ₂ incubator for 72 hours. After the incubation, according to the instructions of the CellTiter-Glo kit (Promega, G7572), 50 μL of pre-melted and equilibrated CTG solution to room temperature was added to each well, mixed with a microplate shaker for 2 minutes, and placed at room temperature for 10 minutes before measuring the fluorescence signal value with an enzyme marker (Envision2104). The cell survival rate (Surviving cells%) data were processed using formula (2), and GraphPad Prism 5.0 software was used to draw an S-shaped dose-survival rate curve using a nonlinear regression model and calculate the IC ₅₀ value. Where V _sample is the reading of the drug-treated group, and V _{vehicle control} is the reading of the control group. Surviving cells% = V _sample / V _{vehicle control} x100% (Formula 2)

Table 10 Proliferation inhibitory activity against NCI-H1975 EGFR-L858R-T790M-C797S cells Compound No. IC ₅₀ (μM) Compound 1 0.112

Conclusion: The compound has good proliferation inhibitory activity against NCI-H1975 EGFR-L858R-T790M-C797S cells.

Test Example 3: Pharmacokinetics test in rats

1.1 Experimental animals: Male SD rats, about 220 g, 6-8 weeks old, 3 rats/compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

1.2 Experimental design: On the day of the experiment, SD rats were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours one day before drug administration and were fed 4 hours after drug administration.

Medication Information Group quantity Medication Information male Test compound Dosage (mg/kg) Dosage concentration (mg/mL) Dosage volume (mL/kg) Collect samples How to give medicine G1 3 Dibenzenesulfonate of compound 1 (amorphous) 10(calculated as free alkali) 1 10 Plasma Oral gavage G2 3 Diphosphate of compound 1 (amorphous form) 10(calculated as free alkali) 1 10 Plasma Oral gavage G3 3 Compound 1 dihydrochloride (amorphous) 10(calculated as free alkali) 1 10 Plasma Oral gavage G4 3 Compound 1, di-methanesulfonate (amorphous form) 10(calculated as free alkali) 1 10 Plasma Oral gavage

Note: Intragastric administration solvent: 0.5% MC (MC: methylcellulose)

Before and after drug administration, 0.15 ml of blood was collected from the eye sockets under isoflurane anesthesia, placed in an EDTAK2 centrifuge tube, and centrifuged at 5000rpm, 4 ^o C for 10 minutes to collect plasma. The blood sampling time points for the intravenous group and the gavage group were: 0, 15, 30min, 1, 2, 4, 6, 8, 24 h. Before analysis and detection, all samples were stored at -80 ^o C and quantitatively analyzed by LC-MS/MS.

Table 11 Pharmacokinetic parameters in rats Test compound How to give medicine AUC _0-t (hr*ng/mL) Dibenzenesulfonate of compound 1 (amorphous) ig (10 mg/kg) 7083 Diphosphate of compound 1 (amorphous form) ig (10 mg/kg) 8324 Compound 1 dihydrochloride (amorphous) ig (10 mg/kg) 8564 Compound 1, di-methanesulfonate (amorphous form) ig (10 mg/kg) 5857

As shown in Table 11, different salts of compound 1 have good oral properties.

Test Example 4: Beagle Dog Pharmacokinetics Test

Experimental animals: Male beagle dogs, about 8-11 kg, 3 per compound, purchased from Beijing Mars Biotechnology Co., Ltd.

Test method: On the day of the test, beagles were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours one day before drug administration, and were fed 4 hours after drug administration. Group quantity Medication Information male Test compound Dosage (mg/kg) Dosage concentration (mg/mL) Dosage volume (mL/kg) Collect samples How to give medicine G1 3 Dibenzenesulfonate of compound 1 (amorphous) 10 2 5 Plasma Oral gavage G2 3 Diphosphate of compound 1 (amorphous form) 10 2 5 Plasma Oral gavage G4 3 Compound 1 dihydrochloride (amorphous) 10 2 5 Plasma Oral gavage G5 3 Compound 1, di-methanesulfonate (amorphous form) 10 2 5 Plasma Oral gavage

Before and after drug administration, 1 ml of blood was collected from the cervical vein or limb vein and placed in an EDTAK2 centrifuge tube. Centrifuge at 5000rpm, 4℃ for 10min, and collect plasma. The blood sampling time points for the gavage group were: 0, 15, 30min, 1, 2, 4, 6, 8, 10, 12, 24, 48 h. Before analysis and testing, all samples were stored at -80℃ and quantitatively analyzed by LC-MS/MS.

Table 12 Pharmacokinetic parameters in dogs Test compound How to give medicine AUC _0-t (hr*ng/mL) Dibenzenesulfonate of compound 1 (amorphous) ig (10 mg/kg) 2611 Diphosphate of compound 1 (amorphous form) ig (10 mg/kg) 2090 Compound 1 dihydrochloride (amorphous) ig (10 mg/kg) 2380 Compound 1, di-methanesulfonate (amorphous form) ig (10 mg/kg) 6090

As shown in Table 12, different salts of compound 1 have good oral properties.

without

FIG1 is an X-ray powder diffraction pattern of maleate salt form 1 of the compound represented by formula (I).

FIG2 is a thermogravimetric analysis spectrum of the maleate salt form 1 of the compound represented by formula (I).

FIG3 is a differential scanning calorimetry curve of the maleate salt form 1 of the compound represented by formula (I).

FIG4 is an isothermal adsorption curve of the maleate salt form 1 of the compound represented by formula (I).

FIG5 is a DVS spectrum of the maleate salt form 1 of the compound represented by formula (I).

FIG6 is an X-ray powder diffraction pattern of the dimaleate crystal form 1 of the compound represented by formula (I).

FIG7 is a thermogravimetric analysis spectrum of the dimaleate crystal form 1 of the compound represented by formula (I).

FIG8 is a differential scanning calorimetry curve of the dimaleate salt form 1 of the compound represented by formula (I).

FIG9 is an isothermal adsorption curve of the dimaleate crystal form 1 of the compound represented by formula (I).

FIG10 is a DVS spectrum of the dimaleate crystal form 1 of the compound represented by formula (I).

FIG11 is an X-ray powder diffraction pattern of 2-naphthalenesulfonic acid form 1 of the compound represented by formula (I).

FIG12 is a thermogravimetric analysis spectrum of 2-naphthalenesulfonic acid form 1 of the compound represented by formula (I).

FIG13 is a differential scanning calorimetry curve of 2-naphthalenesulfonic acid form 1 of the compound represented by formula (I).

FIG14 is an isothermal adsorption curve of 2-naphthalenesulfonic acid form 1 of the compound represented by formula (I).

FIG15 is a DVS spectrum of 2-naphthalenesulfonic acid form 1 of the compound represented by formula (I).

FIG16 is an X-ray powder diffraction pattern of the oxalate salt form 1 of the compound represented by formula (I).

FIG17 is a thermogravimetric analysis spectrum of the oxalate salt form 1 of the compound represented by formula (I).

FIG18 is a differential scanning calorimetry curve of the oxalate salt form 1 of the compound represented by formula (I).

FIG19 is an X-ray powder diffraction pattern of the amorphous benzenesulfonate salt of the compound represented by formula (I).

FIG20 is an X-ray powder diffraction pattern of the amorphous tribenzenesulfonate salt of the compound represented by formula (I).

FIG21 is an X-ray powder diffraction pattern of the amorphous di-L-apple acid salt of the compound represented by formula (I).

FIG22 is an X-ray powder diffraction pattern of the amorphous phosphate of the compound represented by formula (I).

FIG23 is an X-ray powder diffraction pattern of the amorphous diphosphate of the compound represented by formula (I).

FIG24 is an X-ray powder diffraction pattern of the amorphous sulfate of the compound represented by formula (I).

FIG25 is an X-ray powder diffraction pattern of the amorphous disulfate salt of the compound represented by formula (I).

FIG26 is an X-ray powder diffraction pattern of the amorphous trisulfate salt of the compound represented by formula (I).

FIG27 is an X-ray powder diffraction spectrum of the amorphous di-p-toluenesulfonate salt of the compound represented by formula (I).

FIG28 is an X-ray powder diffraction pattern of the amorphous hydrochloride of the compound represented by formula (I).

FIG29 is an X-ray powder diffraction pattern of the amorphous dihydrochloride salt of the compound represented by formula (I).

FIG30 is an X-ray powder diffraction pattern of the amorphous trihydrochloride salt of the compound represented by formula (I).

FIG31 is an X-ray powder diffraction pattern of the amorphous di-2-naphthalenesulfonate of the compound represented by formula (I).

FIG32 is an X-ray powder diffraction pattern of the amorphous hydrobromide salt of the compound represented by formula (I).

FIG33 is an X-ray powder diffraction pattern of the amorphous dihydrobromide salt of the compound represented by formula (I).

FIG34 is an X-ray powder diffraction pattern of the amorphous trihydrobromide salt of the compound represented by formula (I).

FIG35 is an X-ray powder diffraction pattern of the amorphous methanesulfonate salt of the compound represented by formula (I).

FIG36 is an X-ray powder diffraction pattern of the amorphous dimethanesulfonate salt of the compound represented by formula (I).

FIG37 is an X-ray powder diffraction pattern of the amorphous trimesylate salt of the compound represented by formula (I).

FIG38 is an X-ray powder diffraction pattern of the amorphous mandelate salt of the compound represented by formula (I).

FIG39 is an X-ray powder diffraction pattern of the amorphous bis-mandelate salt of the compound represented by formula (I).

FIG40 is an X-ray powder diffraction pattern of the amorphous succinate salt of the compound represented by formula (I).

FIG41 is an X-ray powder diffraction pattern of the amorphous salicylate salt of the compound represented by formula (I).

FIG42 is an X-ray powder diffraction pattern of the amorphous 1,5-naphthalene disulfonic acid salt of the compound represented by formula (I).

FIG43 is an X-ray powder diffraction pattern of the amorphous di-1,5-naphthalene disulfonic acid salt of the compound represented by formula (I).

FIG44 is an X-ray powder diffraction pattern of the amorphous hemifumarate of the compound represented by formula (I).

FIG45 is an X-ray powder diffraction pattern of the amorphous difumarate salt of the compound represented by formula (I).

FIG46 is an X-ray powder diffraction pattern of the amorphous trinicotinate salt of the compound represented by formula (I).

Figure 47 is an X-ray powder diffraction spectrum of the amorphous hippurate salt of the compound represented by formula (I).

Figure 48 is an X-ray powder diffraction pattern of Form 1 of the compound represented by formula (I).

Figure 49 is a thermogravimetric analysis spectrum of Form 1 of the compound represented by formula (I).

Figure 50 is a differential scanning calorimetry curve of Form 1 of the compound represented by formula (I).

without

Claims (18)

A pharmaceutically acceptable salt of a compound represented by formula (I) or a stereoisomer thereof and a solvate thereof, (I) The pharmaceutically acceptable salt is selected from maleate, 2-naphthalenesulfonic acid, 1,5-naphthalene disulfonic acid, fumarate, hydrohalide (preferably hydrobromide and hydrochloride), sulfate, phosphate, L-tartrate, citrate, L-apple acid, hippurate, D-glucuronide, glycolate, mucate, succinate, lactate, orotate, pamoate, glycine, alanine, arginine, cinnamate, benzoate, benzenesulfonate, p-toluenesulfonate, acetate, propionate, valerate, triphenylacetate, L-proline salt, ferulate salt, 2-hydroxyethanesulfonate salt, mandelate salt, nitrate salt, methanesulfonate salt, malonate salt, gentianate salt, salicylate salt, oxalate salt or glutarate salt; preferably selected from benzenesulfonate salt, L-apple acid salt, phosphate salt, sulfate salt, p-toluenesulfonate salt , hydrochloride, maleate, 2-naphthalenesulfonate, hydrobromide, methanesulfonate, citrate, mandelate, lactobionate, succinate, salicylate, 1,5-naphthalenedisulfonate, fumarate, nicotinate, hippurate and oxalate; more preferably methanesulfonate. As described in claim 1, the molar ratio of the compound represented by formula (I): the pharmaceutically acceptable salt is 1:0.5~1:3.5; Preferably, the molar ratio of the compound represented by formula (I): the pharmaceutically acceptable salt is 1:1, 1:2, 1:3. A maleate salt of a compound represented by formula (I) in the form 1, using Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ positions in its X-ray powder diffraction spectrum: 14.83°±0.2°, 16.53°±0.2°, 20.34°±0.2°, 22.87°±0.2°, 23.92°±0.2°; preferably, further at the following 2 The θ position has characteristic diffraction peaks: 4.77°±0.2°, 6.75°±0.2°, 8.80°±0.2°; more preferably, further at the following 2θ positions have characteristic diffraction peaks: 10.97°±0.2°, 16.97°±0.2°, 18.68°±0.2°, 21.08°±0.2°, 24.61°±0.2°. The X-ray powder diffraction pattern of Form 1 as described in claim 3 using Cu-Kα radiation is shown in Figure 1. A dimaleate crystalline form 1 of a compound represented by formula (I) has characteristic diffraction peaks at the following 2θ positions in its X-ray powder diffraction pattern using Cu-Kα radiation: 4.18°±0.2°, 8.24°±0.2°, 18.40°±0.2°, 20.48°±0.2°, 21.96°±0.2°; preferably, further has characteristic diffraction peaks at the following 2θ positions: 18.80°±0. 2°, 23.66°±0.2°, 24.34°±0.2°; more preferably, further having characteristic diffraction peaks at the following 2θ positions: 12.30±0.2°, 13.21°±0.2°, 16.34°±0.2°, 16.57°±0.2°, 20.00°±0.2°, 20.81°±0.2°, 25.73°±0.2°, 27.98°±0.2°. The X-ray powder diffraction pattern of Form 1 as described in claim 5 using Cu-Kα radiation is shown in Figure 6. A 2-naphthalenesulfonate salt of a compound represented by formula (I) in the form 1 has characteristic diffraction peaks at the following 2θ positions using Cu-Kα radiation in its X-ray powder diffraction pattern: 5.82°±0.2°, 17.52°±0.2°, 20.13°±0.2°, 21.16°±0.2°, 26.80°±0.2°; preferably, further has characteristic diffraction peaks at the following 2θ positions: 1 More preferably, the invention further has characteristic diffraction peaks at the following 2θ positions: 7.10°±0.2°, 8.78°±0.2°, 11.59°±0.2°, 17.03°±0.2°, 18.80°±0.2°, 21.72°±0.2°, 22.22°±0.2°. The X-ray powder diffraction pattern of Form 1 as described in claim 7 using Cu-Kα radiation is shown in Figure 11. An oxalate salt of a compound represented by formula (I) in the form 1 has characteristic diffraction peaks at the following 2θ positions using Cu-Kα radiation in its X-ray powder diffraction pattern: 19.69°±0.2°, 20.07°±0.2°, 23.75°±0.2°, 24.45°±0.2°, 26.82°±0.2°; more preferably, further has characteristic diffraction peaks at the following 2θ positions: 13.79 °±0.2°, 17.86°±0.2°, 24.82°±0.2°, 27.07°±0.2°; further, characteristic diffraction peaks are present at the following 2θ positions: 6.91°±0.2°, 7.63°±0.2°, 13.52°±0.2°, 18.87°±0.2°, 20.71°±0.2°, 29.44°±0.2°, 31.28°±0.2°. The X-ray powder diffraction pattern of Form 1 as described in claim 9 using Cu-Kα radiation is shown in Figure 16. The pharmaceutically acceptable salt of the compound represented by formula (I) as described in claim 1 is in the form of an amorphous solid. An amorphous dimethanesulfonate of a compound represented by formula (I). A crystalline form 1 of a compound represented by formula (I), using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ positions: 5.03°±0.2°, 15.35°±0.2°, 19.43°±0.2°, 19.88°±0.2°; preferably, further having characteristic diffraction peaks at the following 2θ positions: 8.03°±0.2°, 13.25°±0.2°, 14.57°±0 .2°, 14.88°±0.2°, 23.83±0.2°, 24.71°±0.2°, 26.44°±0.2°, 29.93°±0.2°; more preferably, further having characteristic diffraction peaks at the following 2θ positions: 8.99°±0.2°, 20.17±0.2°, 22.32±0.2°, 22.55±0.2°, 26.17±0.2°, 29.49±0.2°. The X-ray powder diffraction pattern of Form 1 as described in claim 13 using Cu-Kα radiation is shown in Figure 48. A method for preparing a pharmaceutically acceptable salt of a compound represented by formula (I), wherein the method comprises: a step of forming a salt with the compound represented by formula (I) and an acid. A preparation method as described in claim 15, wherein the solvent used is selected from one or more of C1-6 halogenated alkane solvents, C2-6 ester solvents, C2-6 ether solvents, C1-6 alcohol solvents or water, and preferably the solvent used is selected from one or more of dichloromethane, 1,2-dichloroethane, ethyl acetate, methanol, ethanol, isopropanol, propanol, diethyl ether, tetrahydrofuran and water. A pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of the crystalline form or amorphous form according to any one of claims 1 to 15, and a pharmaceutically acceptable carrier or excipient. A use of a pharmaceutically acceptable salt and a solvate thereof as described in claim 1 or 2, a crystalline form or amorphous form as described in any one of claims 3 to 14, or a pharmaceutical composition as described in claim 17 in the preparation of a drug for treating and inhibiting or degrading EGFR-related diseases, preferably the disease is selected from cancer.

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