P2X3 inhibitor compounds and salts, polymorphs, and uses thereof

Abstract

The present application relates to a kind of P2X3 inhibitor compounds and its salt, polymorph and purpose.The present application provides the crystal form of compound of formula I, with good medicinal property.It is also obtained that the pharmaceutically acceptable salt of compound of formula I, and further obtains the crystal form product of salt, such as hydrochloride salt crystal form A, maleate salt crystal form A, p-toluenesulfonic acid salt crystal form A, benzenesulfonic acid salt crystal form A, malonate salt crystal form A.It is of great significance for the development of effective therapeutic drugs.

Description

[0001] This application claims priority to the following prior applications: patent application No. 202210326113.5, filed with the China National Intellectual Property Administration on March 29, 2022, entitled "A P2X3 Inhibitor Compound and Its Salt, Polymorphs and Uses"; and patent application No. 202310281376.3, filed with the China National Intellectual Property Administration on March 21, 2023, entitled "A P2X3 Inhibitor Compound and Its Salt, Polymorphs and Uses". The entire contents of the aforementioned prior applications are incorporated herein by reference. Technical Field

[0002] This invention belongs to the pharmaceutical field and relates to a P2X3 inhibitor compound, its salts, polymorphs, preparation methods, and applications thereof. Background Technology

[0003] P2X receptors are non-selective ATP-gated ion channel receptors, i.e., purinergic receptors, that bind to extracellular ATP, primarily derived from damaged or inflamed tissues. These receptors are widely expressed in the nervous, immune, cardiovascular, skeletal, gastrointestinal, respiratory, and endocrine systems, and participate in various physiological processes, including regulation of heart rhythm and contractility, regulation of vascular tone, nociception (especially chronic pain), vas deferens contraction during ejaculation, bladder contraction during urination, platelet aggregation, macrophage activation, apoptosis, and neuron-glial interactions. The aforementioned P2X receptors include seven homologous receptors: P2X1, P2X2, P2X3, P2X4, P2X5, P2X6, and P2X7, and three heterologous receptors: P2X2 / 3, P2X4 / 6, and P2X1 / 5.

[0004] P2X3 is a subtype of the P2X receptor family that is selectively expressed in dorsal root ganglia of nerve endings, spinal cord, and brain neurons, specifically in small to medium diameter primary sensory neurons.

[0005] Numerous studies have shown that activation of P2X3 and P2X2 / 3 expressed in primary sensory neurons plays a crucial role in acute injury, hyperalgesia, and hypersensitivity responses in rodents. Many studies have demonstrated that upregulation of P2X3 receptor expression can lead to hyperalgesia and participate in pain signaling. P2X3 knockout mice exhibit reduced pain responses, and P2X3 receptor antagonists have shown a role in reducing nociception in pain and inflammatory pain models.

[0006] P2X3 is distributed in primary afferent nerves surrounding the airways and can regulate cough. Studies have shown that ATP released from damaged or inflamed airway tissue acts on P2X3 receptors in primary neurons, triggering depolarization and action potentials. These potentials transmit cough impulses, initiating coughing. P2X3 receptors play an important role in the cough reflex hypersensitivity response; by antagonizing P2X3 receptor binding, the cough reflex hypersensitivity response can be inhibited, thereby suppressing excessive coughing in patients with chronic cough. Furthermore, research has shown that P2X3 antagonists can treat chronic obstructive pulmonary disease, pulmonary fibrosis, pulmonary hypertension, or asthma; therefore, P2X3 antagonists also hold promise as new drugs for treating these diseases.

[0007] P2X3 has been reported to be involved in the afferent pathway controlling the bladder capacity reflex; P2X3 knockout mice exhibit significantly reduced urination frequency and significantly increased bladder capacity. Therefore, inhibiting the binding of P2X3 receptor antagonists to the P2X3 receptor could be effective in treating conditions involving urinary storage and voiding disorders, such as overactive bladder. Thus, P2X3 antagonists may be potential drugs for treating overactive bladder and related diseases.

[0008] P2X3 antagonists show great promise. Currently, commonly used cough medications such as gabapentin, morphine, and amitriptyline, or treatments using speech pathology, can improve coughs in many patients, but they are not suitable for all patients. Moreover, centrally acting drugs such as gabapentin may produce adverse side effects and are not suitable for long-term use. There is an urgent clinical need to develop long-term medications for chronic refractory coughs to provide doctors with treatment options. Therefore, the development of P2X3 antagonists is of great clinical significance.

[0009] Chinese patent application CN202111165441.3 discloses the structure of compound I:

[0010]

[0011] Compound I can effectively antagonize P2X3 receptor activity and has broad application prospects in the preparation of drugs for treating P2X3-related diseases. Therefore, further research on compound I and its salt form and crystal form is of great significance for the development of effective therapeutic drugs. Summary of the Invention

[0012] To address the problems existing in the prior art, the present invention provides a crystal form of a compound of formula I or a pharmaceutically acceptable salt thereof, wherein the structure of the compound of formula I is as follows:

[0013]

[0014] In some embodiments, the present invention provides a free basal crystal form A of the compound of Formula I, wherein the X-ray powder diffraction pattern of the free basal crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.44°, 14.87°, 15.77°, 17.81°, and 18.61°; further, the X-ray powder diffraction pattern of the free basal crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.44°, 11.14°, 11.36°, 14.87°, 15.77°, 16.97°, 17.81°, and 18.61°. A diffraction peak is observed at 1°; furthermore, the X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 3.75°, 7.44°, 11.14°, 11.36°, 11.98°, 12.25°, 14.87°, 15.77°, 16.97°, 17.81°, 18.61°, and 22.36°; even further, the X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 3.75°, 5.99°, 7.44°, 9.99°, 11.36 ... Diffraction peaks are observed at 0.01°, 9.93°, 11.14°, 11.36°, 11.98°, 12.25°, 13.88°, 14.20°, 14.87°, 15.77°, 16.97°, 17.81°, 18.61°, 22.36°, and 24.07°. Furthermore, the X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows peaks at 3.75°, 5.99°, 7.44°, 9.01°, 9.93°, 11.14°, 11.36°, 1... Diffraction peaks are observed at 1.98°, 12.25°, 13.88°, 14.20°, 14.87°, 15.77°, 16.97°, 17.81°, 18.61°, 19.39°, 20.26°, 21.14°, 22.36°, 23.34°, 24.07°, 26.33°, 26.78°, 27.18°, 28.17°, 30.20°, 33.88°, 34.35°, 37.23°, and 37.70°; furthermore, the free alkali crystal form A has essentially the following characteristics... Figure 1-1 The XRPD spectrum shown.

[0015] In some embodiments, the free alkali crystal form A has one, two, or three of the following characteristics:

[0016] (1) The TGA curve of free alkali crystal form A shows a weight loss of approximately 1.28±1% at 150.0±3℃;

[0017] (2) The DSC curve of free alkali crystal form A has an endothermic peak at 175.6±3℃.

[0018] (3) The DSC curve of free alkali crystal form A has an endothermic peak at 176.4±3℃.

[0019] In some embodiments, the TGA / DSC chart of the free alkali crystal form A is as follows: Figure 1-2 As shown; the free alkali crystal form A 1 H NMR image as follows Figure 1-3 As shown.

[0020] According to an embodiment of the present invention, the free alkali crystal form A is an amorphous form.

[0021] In some embodiments, the free alkali crystal form A is in the form of aggregated needle-like crystals.

[0022] In some embodiments, the present invention provides a free basal crystal form B of the compound of Formula I, wherein the X-ray powder diffraction pattern of the free basal crystal form B, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.21°, 12.48°, 13.17°, 14.41°, 19.09°, 19.56°, 22.09°, and 26.49°; further, the X-ray powder diffraction pattern of the free basal crystal form B, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.21°, 12.48°, 13.17°, 14.41°, 19.09°, 19.56°, 22.09°, and 26.49°. Diffraction peaks are observed at 0.48°, 13.17°, 14.41°, 16.72°, 19.09°, 19.56°, 20.90°, 22.09°, and 26.49°. Furthermore, the X-ray powder diffraction pattern of the free alkali crystal form B, expressed as a diffraction angle of 2θ±0.2°, shows peaks at 7.21°, 8.35°, 12.48°, 13.17°, 14.41°, 15.05°, 16.72°, 17.80°, 18.39°, and 19.49°. Diffraction peaks are observed at 09°, 19.56°, 20.90°, 21.67°, 22.09°, 22.97°, 25.16°, 26.49°, and 27.49°. Furthermore, the X-ray powder diffraction pattern of the free alkali crystal form B, expressed as a diffraction angle of 2θ ± 0.2°, shows peaks at 7.21°, 8.35°, 12.48°, 13.17°, 14.41°, 15.05°, 16.72°, 17.80°, 18.39°, and 19.49°. Diffraction peaks are observed at 09°, 19.56°, 20.90°, 21.67°, 22.09°, 22.97°, 25.16°, 25.45°, 26.49°, 27.49°, 28.66°, 29.10°, 29.35°, 31.71°, 32.00°, 32.85°, 33.70°, 34.23°, 36.78°, 38.26°, and 38.70°; furthermore, the free alkali crystal form B has essentially the following characteristics... Figure 2-1 The XRPD spectrum shown.

[0023] In some embodiments, the free alkali crystal form B has one, two, or three of the following characteristics:

[0024] (1) The TGA curve of free alkali crystal form B shows a weight loss of approximately 2.36±1% at 150.0±3℃;

[0025] (2) The DSC curve of free alkali crystal form B has an endothermic peak at the starting point of 177.0±3℃;

[0026] (3) The DSC curve of free alkali crystal form B has an endothermic peak at 179.4±3℃.

[0027] In some embodiments, the TGA / DSC plot of the free alkali crystal form B is as follows: Figure 2-2 As shown; the free alkali crystal form B 1 H NMR image as follows Figure 2-3 As shown.

[0028] According to an embodiment of the present invention, the free alkali crystal form B is an amorphous form.

[0029] On the other hand, the present invention provides pharmaceutically acceptable salts of the compound of Formula I, which may be selected from salts formed by the compound of Formula I with inorganic or organic acids, such as: hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, hydrobromic acid; for example, the organic acids include: maleic acid, L-aspartic acid, fumaric acid, L-tartaric acid, citric acid, D-glucuronic acid, L-malic acid, hippuric acid, D-gluconic acid, DL-lactic acid, succinic acid, L-ascorbic acid, adipic acid, acetic acid, p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, oxalic acid, 2-hydroxyethanesulfonic acid, malonic acid, gentian acid, benzoic acid;

[0030] According to a preferred embodiment of the present invention, the pharmaceutically acceptable salt of the compound of formula I is the hydrochloride, maleate, p-toluenesulfonate, benzenesulfonate or malonate of the compound of formula I.

[0031] According to embodiments of the present invention, those skilled in the art will understand that when the compound of formula I forms a salt with an acid, the molar ratio of the compound of formula I to the acid can be 5:1 to 1:5, for example 3:1, 2:1, 1:1, 1:1.5, 1:2, 1:2.5, or 1:3. Preferably, the molar ratio of the compound of formula I to the acid is 1:1 or 2:1.

[0032] On the other hand, the present invention provides a crystal form of a pharmaceutically acceptable salt of the compound of formula I.

[0033] In some embodiments, the present invention provides a hydrochloride crystal form A of the compound of Formula I, wherein the X-ray powder diffraction pattern of the hydrochloride crystal form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.77°, 9.01°, 10.10°, 15.54°, 17.51°, 19.24°, and 24.49°; further, the X-ray powder diffraction pattern of the hydrochloride crystal form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.77°, 9.01°, 10.10°, 15.54°, 17.51°, 18.01°, 19.24°, 20.05°, 21.28°, and 23. Diffraction peaks are observed at 38°, 23.79°, 24.49°, 26.07°, and 28.33°. Further, the X-ray powder diffraction pattern of the hydrochloride crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.77°, 9.01°, 10.10°, 15.54°, 17.51°, 18.01°, 19.24°, 20.05°, 21.28°, 21.59°, 22.67°, 23.38°, 23.79°, 24.49°, 26.07°, 27.17°, and 28.33°. Even further, the hydrochloride crystal form A has essentially the following characteristics... Figure 3-1 The XRPD spectrum shown.

[0034] In some embodiments, the molar ratio of the hydrochloride crystal form A, compound of formula I, to hydrochloric acid is 2:1.

[0035] According to an embodiment of the present invention, the hydrochloride crystal form A has a substantially as follows Figure 9-4 The VT-XRPD spectrum is shown.

[0036] According to an embodiment of the present invention, the hydrochloride crystal form A is amorphous.

[0037] In some embodiments, the hydrochloride crystal form A has one, two, three, four, five or six of the following characteristics: (1) the TGA curve of the hydrochloride crystal form A shows a weight loss of about 2.19±1% ​​at 100.0±3℃;

[0038] (2) The TGA curve of hydrochloride crystal form A shows a weight loss of approximately 3.90±1% in the temperature range of 100.0±3℃ to 160.0±3℃;

[0039] (3) The DSC curve of hydrochloride crystal form A has an endothermic peak at the starting point of 143.6±3℃;

[0040] (4) The DSC curve of hydrochloride crystal form A has an endothermic peak at 157.4±3℃;

[0041] (5) The DSC curve of hydrochloride crystal form A has an endothermic peak at 176.0±3℃;

[0042] (6) The DSC curve of hydrochloride crystal form A has an endothermic peak at 179.0±3℃.

[0043] In some embodiments, the TGA / DSC plot of the hydrochloride crystal form A is as follows: Figure 9-2 As shown; the hydrochloride crystal form A 1 H NMR image as follows Figure 3-2 As shown.

[0044] In some embodiments, no solvent residue is present in the hydrochloride crystal form A.

[0045] In some embodiments, the hydrochloride crystal form A is in the form of irregular granules.

[0046] In some embodiments, the present invention provides a maleate crystal form A of the compound of Formula I, wherein the X-ray powder diffraction pattern of the maleate crystal form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 6.73°, 10.84°, 14.68°, 16.26°, 18.23°, and 18.44°; further, the X-ray powder diffraction pattern of the maleate crystal form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.43°, 6.73°, 10.84°, 14.68°, 16.26°, 16.82°, 18.23°, and 18.44°. A diffraction peak is observed at 8.44°; furthermore, the X-ray powder diffraction pattern of the maleate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.43°, 6.73°, 9.95°, 10.84°, 11.75°, 13.50°, 14.68°, 16.26°, 16.82°, 18.23°, 18.44°, 20.17°, 22.79°, 23.22°, 24.00°, 26.07°, 27.72°, and 28.86°; even further, the maleate crystal form A has essentially the following characteristics... Figure 4-1 The XRPD spectrum shown.

[0047] In some embodiments, the molar ratio of the maleate crystal form A (Form I compound) to maleic acid is 2:1.

[0048] According to an embodiment of the present invention, the maleate crystal form A has essentially the following properties: Figure 10-4 The VT-XRPD spectrum is shown.

[0049] According to an embodiment of the present invention, the maleate crystal form A is an amorphous form.

[0050] In some embodiments, the maleate crystal form A has one, two, three or more of the following characteristics:

[0051] (1) The TGA curve of maleate crystal form A shows a weight loss of approximately 1.65±1% at 110.0±3℃;

[0052] (2) The TGA curve of maleate crystal form A shows a weight loss of approximately 11.88±1% in the temperature range of 110.0±3℃ to 220.0±3℃;

[0053] (3) The DSC curve of maleate crystal form A has an endothermic peak at 107.8±3℃;

[0054] (4) The DSC curve of maleate crystal form A has an endothermic peak starting point at 143.4±3℃;

[0055] (5) The DSC curve of maleate crystal form A has an endothermic peak at 144.1±3℃;

[0056] (6) The DSC curve of maleate crystal form A has an endothermic peak at 160.2±3℃.

[0057] In some embodiments, the TGA / DSC plot of the maleate crystal form A is as follows: Figure 10-2 As shown; the maleate crystal form A 1 H NMR image as follows Figure 4-2 As shown.

[0058] In some embodiments, the present invention provides a p-toluenesulfonate crystal form A of the compound of Formula I, wherein the X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 4.99°, 7.26°, 8.70°, 8.87°, 15.40°, 17.73°, 21.01°, and 24.13°; further, the X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 4.99°, 7.26°, 8.70°, 8.87°, 15.40°, 17.73°, 21.01°, and 24.13°. Diffraction peaks were observed at 6°, 8.70°, 8.87°, 15.20°, 15.40°, 16.68°, 17.73°, 19.71°, 21.01°, and 24.13°. Furthermore, the X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ ± 0.2°, showed peaks at 4.99°, 7.26°, 8.70°, 8.87°, 14.45°, 14.88°, 15.20°, 15.40°, 16.41°, and 16.68°. Diffraction peaks were observed at 17.45°, 17.73°, 19.16°, 19.71°, 20.66°, 21.01°, 21.76°, 22.41°, 24.13°, 25.76°, 26.18°, and 27.25°. Furthermore, the X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ ± 0.2°, showed peaks at 4.99°, 7.26°, 8.70°, 8.87°, 14.45°, 14.88°, and 15.20°. Diffraction peaks are observed at 15.40°, 16.41°, 16.68°, 17.45°, 17.73°, 19.16°, 19.71°, 20.66°, 21.01°, 21.76°, 22.41°, 24.13°, 25.76°, 26.18°, 27.25°, 27.95°, 29.23°, 30.69°, 31.00°, 31.78°, and 38.41°; furthermore, the p-toluenesulfonate crystal form A has essentially the following characteristics... Figure 5-1 The XRPD spectrum shown.

[0059] In some embodiments, the molar ratio of the p-toluenesulfonate crystal form A (formula I) to p-toluenesulfonic acid is 1:1.

[0060] According to an embodiment of the present invention, the p-toluenesulfonate crystal form A is an amorphous form.

[0061] In some embodiments, the p-toluenesulfonate crystal form A has one, two, or three of the following characteristics:

[0062] (1) The TGA curve of p-toluenesulfonate crystal form A shows a weight loss of approximately 0.73±1% at 150.0±3℃;

[0063] (2) The DSC curve of p-toluenesulfonate crystal form A has an endothermic peak starting point at 157.1±3℃;

[0064] (3) The DSC curve of p-toluenesulfonate crystal form A has an endothermic peak at 159.2±3℃.

[0065] In some embodiments, the TGA / DSC plot of the p-toluenesulfonate crystal form A is as follows: Figure 11-2 As shown; the p-toluenesulfonate crystal form A 1 H NMR image as follows Figure 5-2 As shown.

[0066] In some embodiments, the present invention provides a benzenesulfonate crystal form A of the compound of formula I, wherein the X-ray powder diffraction pattern of the benzenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.36°, 7.28°, 8.34°, 9.64°, 16.20°, 18.55°, and 21.49°; further, the X-ray powder diffraction pattern of the benzenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 5.36°, 7.28°, 8.34°, 9.64°, 16.20°, 18.55°, 19.28°, 21.49°, 21.81°, 23.21°, 25.05°, and 25. A diffraction peak is observed at 74°; furthermore, the X-ray powder diffraction pattern of the benzenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.36°, 7.28°, 8.34°, 9.64°, 10.66°, 14.55°, 15.00°, 16.20°, 16.93°, 17.85°, 18.55°, 19.28°, 19.74°, 20.80°, 21.49°, 21.81°, 23.21°, 23.68°, 23.98°, 25.05°, 25.74°, 26.65°, and 27.82°; even further, the benzenesulfonate crystal form A has essentially the following characteristics... Figure 6-1 The XRPD spectrum shown.

[0067] In some embodiments, the molar ratio of the benzenesulfonate crystal form A, compound of formula I, to benzenesulfonic acid is 1:1.

[0068] According to an embodiment of the present invention, the benzenesulfonate crystal form A is an amorphous form.

[0069] In some embodiments, the benzenesulfonate crystal form A has one, two, or three of the following characteristics:

[0070] (1) The TGA curve of benzenesulfonate crystal form A shows a weight loss of approximately 1.35±1% at 120.0±3℃;

[0071] (2) The DSC curve of benzenesulfonate crystal form A has an endothermic peak at the starting point of 159.6±3℃;

[0072] (3) The DSC curve of benzenesulfonate crystal form A has an endothermic peak at 160.9±3℃.

[0073] In some embodiments, the TGA / DSC plot of the benzenesulfonate crystal form A is as follows: Figure 6-2 As shown; the benzenesulfonate crystal form A 1 H NMR image as follows Figure 6-3 As shown.

[0074] In some embodiments, the present invention provides a malonate crystal form A of the compound of Formula I, wherein the X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 6.75°, 9.96°, 10.67°, 14.48°, 16.04°, 16.88°, 18.04°, and 18.29°; further, the X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 6.75°, 9.96°, 10.67°, 14.48°, 16.04°, 16.88°, 18.04°, and 18.29°. The diffraction pattern shows diffraction peaks at 5.34°, 6.75°, 9.96°, 10.67°, 11.83°, 13.49°, 14.48°, 16.04°, 16.88°, 17.04°, 18.04°, 18.29°, and 27.38°. Furthermore, the X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows peaks at 5.34°, 6.75°, 9.96°, 10.67°, 11.83°, 13.49°, 14.48°, 16.04°, 16.88°, 17.04°, 18.04°, 18.29°, and 27.38°. Diffraction peaks were observed at 67°, 11.83°, 13.49°, 14.48°, 16.04°, 16.88°, 17.04°, 18.04°, 18.29°, 20.27°, 22.57°, 22.95°, 27.38°, and 28.83°. Furthermore, the X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, showed peaks at 5.34°, 6.75°, 9.96°, and 28.83°. Diffraction peaks are observed at 10.67°, 11.83°, 13.49°, 14.48°, 16.04°, 16.88°, 17.04°, 18.04°, 18.29°, 18.63°, 20.27°, 21.57°, 22.57°, 22.95°, 24.11°, 24.83°, 26.02°, 27.38°, and 28.83°; furthermore, the malonate crystal form A has essentially the following characteristics... Figure 7-1 The XRPD spectrum shown.

[0075] In some embodiments, the molar ratio of the malonate crystal form A (form I compound) to malonic acid is 2:1.

[0076] According to an embodiment of the present invention, the malonate crystal form A has essentially the following properties: Figure 7-4 The VT-XRPD spectrum is shown.

[0077] According to an embodiment of the present invention, the malonate crystal form A is amorphous.

[0078] In some embodiments, the malonate crystal form A has one, two, three, four, or five of the following characteristics:

[0079] (1) The TGA curve of malonate crystal form A shows a weight loss of approximately 2.99±1% at 120.0±3℃;

[0080] (2) The TGA curve of malonate crystal form A shows a weight loss of approximately 11.02±1% in the temperature range of 120.0±3℃ to 200.0±3℃;

[0081] (3) The DSC curve of malonate crystal form A has an endothermic peak at 155.6±3℃.

[0082] (4) The DSC curve of malonate crystal form A has an endothermic peak at 156.4±3℃;

[0083] (5) The DSC curve of malonate crystal form A has an endothermic peak at 172.4±3℃.

[0084] In some embodiments, the TGA / DSC plot of the malonate crystal form A is as follows: Figure 7-2 As shown; the malonate crystal form A 1 H NMR image as follows Figure 7-3 As shown.

[0085] On the other hand, the present invention provides a method for preparing the free base crystal form A of the compound of formula I, which includes the following methods:

[0086] Method 1: Add compound I to organic solvent I, dissolve, filter, and volatilize at room temperature.

[0087] Preferably, the organic solvent I is selected from one or more of acetone, tetrahydrofuran, dichloromethane, acetonitrile, and ethyl acetate;

[0088] Method 2: Dissolve the compound of formula I completely in organic solvent II, and add the antisolvent dropwise to the clear solution under stirring until a solid precipitates; if no solid precipitates, use suspension stirring; if no solid still precipitates, cool down and suspend stirring, then allow the clear solution to evaporate at room temperature.

[0089] The organic solvent II may be selected from one or more of methanol, acetone, ethyl acetate, tetrahydrofuran, chloroform, N,N-dimethylacetamide, and N-methylpyrrolidone;

[0090] The antisolvent may be selected from one or more of water, m-xylene, n-hexane, cumene, toluene, cyclohexane, n-heptane, n-pentane, and p-isopropyltoluene, for example, selected from water, mixtures of water and m-xylene, mixtures of water and n-hexane, mixtures of water and cumene, mixtures of water and toluene, and mixtures of cyclohexane and p-isopropyltoluene.

[0091] Method 3: Place the first sample vial containing compound I openly into a second sample vial containing solvent, seal the second sample vial, and let it stand at room temperature; the solvent should not cover the mouth of the first sample vial.

[0092] The solvent is selected from one or more of ethanol, dichloromethane, acetonitrile, acetone, toluene, N,N-dimethylacetamide, and n-hexane;

[0093] Preferably, the settling time is 1 to 8 days.

[0094] Method 4: Place the first sample vial containing the solution of compound I openly into the second sample vial containing the antisolvent, seal the second sample vial, and let it stand at room temperature; the antisolvent does not submerge the mouth of the first sample vial.

[0095] The solvent in the compound solution of Formula I is selected from one or more of isopropanol, methyl isobutyl ketone, 1,4-dioxane and dimethyl sulfoxide, preferably dimethyl sulfoxide;

[0096] The antisolvent is selected from one or more of n-pentane, methyl butyl ether, water and m-xylene, preferably a mixture of n-pentane and methyl butyl ether, or a mixture of water and m-xylene.

[0097] Method 5: Add the polymer to the solution of compound I and allow it to evaporate at room temperature;

[0098] The solvent in the compound solution of Formula I is selected from one or more of methanol, 2-butanone, methyl acetate, isopropyl acetate, ethanol, dichloromethane, and 2-methyltetrahydrofuran;

[0099] The polymer is selected from one or more of polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose, methylcellulose, polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate, and hydroxyethyl cellulose.

[0100] Preferably, the polymer is selected from mixed polymer A or mixed polymer B; mixed polymer A is composed of polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose and methylcellulose, and mixed polymer B is composed of polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate and hydroxyethyl cellulose;

[0101] Preferably, the mass ratio of the polymer to the compound of formula I is 1:(5-20).

[0102] Method 6: Place the first sample vial containing compound I openly into a second sample vial containing a saturated salt solution or water, seal the second sample vial, and let it stand at room temperature; the solvent should not cover the mouth of the first sample vial.

[0103] Preferably, the saturated salt solution is an inorganic salt saturated solution, such as a saturated potassium acetate solution, a saturated potassium carbonate solution, a saturated sodium bromide solution, or a saturated potassium bromide solution.

[0104] Preferably, the humidity of the system is 15–100% RH.

[0105] Method 7: The suspension of compound I is stirred and centrifuged under temperature cycling, and the solid is collected;

[0106] The solvent in the suspension is selected from one or more of n-heptane, methyl butyl ether, anisole, dicyclohexylamine, acetone, ethanol, ethyl acetate, methylcyclohexane, chloroform, 2-butanone, m-xylene, and water, for example, a mixture of n-heptane, methyl butyl ether, anisole, butanone, and water, a mixture of ethanol and water, a mixture of ethyl acetate and methylcyclohexane, a mixture of chloroform and n-heptane, or a mixture of 2-butanone and m-xylene;

[0107] Preferably, the temperature cycling conditions include: 50℃~5℃, 0.1~0.5℃ / min, and at least 2 cycles.

[0108] Method 8: Dissolve the compound of formula I completely in the positive solvent, and add the antisolvent dropwise to the clear solution under stirring until a solid precipitates; if no solid precipitates, use suspension stirring; if no solid still precipitates, cool down and suspend stirring, then allow the clear solution to evaporate at room temperature;

[0109] The solvent may be selected from one or more of ethanol, ethyl acetate, 2-methyltetrahydrofuran, 2-butanone, acetonitrile, dichloromethane, and 1,4-dioxane;

[0110] The antisolvent is selected from one or more of n-heptane, tetrahydrofuran, and water.

[0111] Method 9: Place the turbid liquid of compound I at room temperature, stir magnetically, centrifuge, and collect the solid;

[0112] The solvent of the turbid liquid is selected from one or more of isobutanol, methyl tert-butyl ether, cyclohexane, toluene, isopropyl acetate, water, methylcyclohexane, tetrahydrofuran, n-pentane, acetone, isopropanol, cyclopentyl methyl ether, methanol, p-isopropyltoluene, dichloromethane, n-heptane, acetonitrile, 1,4-dioxane, and N-methylpyrrolidone; for example, the solvent of the turbid liquid is selected from a mixture of isobutanol, methyl tert-butyl ether, cyclohexane, toluene, isopropyl acetate, and toluene, water, a mixture of methylcyclohexane, tetrahydrofuran, and n-pentane, a mixture of acetone and isopropanol, a mixture of isopropanol and cyclopentyl methyl ether, a mixture of dichloromethane and n-heptane, a mixture of acetonitrile and water, a mixture of 1,4-dioxane and water, or a mixture of N-methylpyrrolidone and water;

[0113] Preferably, the magnetic stirring speed is 700-1200 rpm.

[0114] Method 10: Weigh the compound of Formula I into an HPLC bottle, add solvent to the HPLC bottle, heat and stir to equilibrate, then filter to obtain the supernatant; place the supernatant in a biological incubator, cool it from 50°C to 5°C at a rate of 0.05°C / min, and then maintain the temperature at 5°C; transfer the clear solution to -20°C for constant temperature, collect the precipitated solid, and transfer the sample without precipitated solid to room temperature for volatilization;

[0115] The solvent is selected from one or more of isopropanol, anisole, isopropyl acetate, tetrahydrofuran, and water;

[0116] The heating temperature is 45-55℃, preferably 50℃.

[0117] Method 11: Stir the suspension of compound I, centrifuge, and collect the solid;

[0118] The solvent in the suspension is selected from one or more of the following: n-butanol (n-BuOH), toluene, diisopropyl ether, methylcyclohexane, cumene, anisole, water, petroleum ether, dicyclohexylamine, 2-methyltetrahydrofuran, n-hexane, 2-butanone, isopropyl acetate, chloroform, m-xylene, tetrahydrofuran, methyl isobutyl ketone, cyclopentyl methyl ether, and benzyl alcohol; for example, a mixture of n-butanol and toluene, diisopropyl ether, methylcyclohexane, cumene, anisole, water, PET, dicyclohexylamine, a mixture of 2-butanone and cumene, a mixture of isopropyl acetate and toluene, a mixture of chloroform and m-xylene, a mixture of isopropyl acetate and water, a mixture of tetrahydrofuran and water, a mixture of methyl isobutyl ketone and cyclopentyl methyl ether, or a mixture of benzyl alcohol and toluene;

[0119] The suspension is stirred at 45-55°C, preferably at 50°C;

[0120] Preferably, the stirring is magnetic stirring, for example, the magnetic stirring speed is 700-1200 rpm.

[0121] On the other hand, the present invention provides a method for preparing the free base crystal form B of the compound of formula I, which includes the following steps:

[0122] The free acid crystal form A of the compound of formula I was dissolved in 1,4-dioxane and then subjected to gas-liquid diffusion in a hexane atmosphere to obtain the free base crystal form B of the compound of formula I.

[0123] The present invention also provides a method for preparing a pharmaceutically acceptable salt of the compound of formula I, comprising the steps of: mixing the compound of formula I or the free base crystal form A of the compound of formula I with a salt-forming agent (e.g., the corresponding acid) in a suitable solvent to obtain a mixture.

[0124] In some embodiments, the preparation method further includes the steps of: stirring or pulping the mixture, separating the solids, and vacuum drying to obtain a pharmaceutically acceptable salt of the compound of formula I; preferably, the stirring, pulping, and vacuum drying are carried out at room temperature.

[0125] In some embodiments, the preparation method further includes the step of increasing the supersaturation of the mixture (e.g., by adding an antisolvent).

[0126] In some embodiments, the solvent is selected from one or a mixture of several of ethanol, heptane, ethyl acetate, MTBE, acetonitrile, water, and acetone.

[0127] In another aspect, the present invention provides a pharmaceutical composition comprising one or more of the following: a free base crystal form of the compound of formula I (e.g., free base crystal form A, free base crystal form B) and a pharmaceutically acceptable salt of the compound of formula I (including its crystal form).

[0128] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient or carrier.

[0129] In another aspect, the present invention provides the use of the free base crystal form of the compound of formula I (e.g., free base crystal form A, free base crystal form B), a pharmaceutically acceptable salt of the compound of formula I (including its crystal form), or the pharmaceutical composition thereof in the preparation of a medicament for the treatment and / or prevention of P2X3-related diseases.

[0130] According to embodiments of the present invention, using the free base crystal form of the compound of formula I (e.g., free base crystal form A, free base crystal form B), a pharmaceutically acceptable salt of the compound of formula I (including its crystal form), or the pharmaceutical composition thereof, better and more effective clinical treatment drugs or regimens can be provided to patients in need.

[0131] The present invention also provides a method for treating and / or preventing diseases related to P2X3, the method comprising administering to a patient a therapeutically effective dose of a crystal form comprising a compound of formula I as described above, a pharmaceutically acceptable salt of a compound of formula I as described above, a crystal form of a salt as described above, or a pharmaceutical composition as described above; preferably, a pharmaceutical formulation comprising a free base crystal form of a compound of formula I as described above (e.g., free base crystal form A, free acid crystal form B), a pharmaceutically acceptable salt of a compound of formula I (including its crystal form), or the pharmaceutical composition described above.

[0132] In some preferred embodiments, the pharmaceutically acceptable salt of the Formula I compound comprises a salt formed by the Formula I compound and an acid selected from the following: hydrochloric acid, sulfuric acid, maleic acid, L-aspartic acid, phosphoric acid, fumaric acid, L-tartaric acid, citric acid, D-glucuronic acid, L-malic acid, hippuric acid, D-gluconic acid, DL-lactic acid, succinic acid, L-ascorbic acid, adipic acid, acetic acid, p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, oxalic acid, 2-hydroxyethanesulfonic acid, malonic acid, gentian acid, and benzoic acid.

[0133] Preferably, the pharmaceutically acceptable salt of the compound of Formula I includes the hydrochloride crystal form A, maleate crystal form A, p-toluenesulfonate crystal form A, benzenesulfonate crystal form A, malonate crystal form A, or any combination of multiple salts thereof.

[0134] According to an embodiment of the present invention, the P2X3-related diseases include: pain, reproductive and urinary system diseases, or respiratory system diseases.

[0135] Preferably, the pain includes: inflammatory pain, surgical pain, visceral pain, toothache, premenstrual pain, central pain, burn-related pain, migraine, or cluster headache; preferably, the reproductive and urinary system diseases include: urinary incontinence, overactive bladder, dysuria, cystitis, endometriosis, and endometriosis-related pain; preferably, the respiratory system diseases include: cough, idiopathic pulmonary fibrosis (IPF), and chronic obstructive pulmonary disease (COPD); preferably, the cough includes subacute or chronic cough, treatment-resistant cough, idiopathic chronic cough, post-viral cough, iatrogenic cough, and cough associated with respiratory diseases (e.g., COPD, asthma, and bronchospasm).

[0136] According to an embodiment of the present invention, the P2X3-related diseases include at least one of the following diseases: chronic cough, especially refractory chronic cough (RCC) and chronic cough of unknown cause (UCC).

[0137] The present invention also provides a method for quality determination of the crystal form of a compound of formula I or a pharmaceutically acceptable salt thereof, comprising the following steps: determining the content of the crystal form using high performance liquid chromatography; wherein the mobile phase used in the high performance liquid chromatography includes mobile phase A and mobile phase B;

[0138] The mobile phase A is an aqueous solution of formic acid (FA) and acetonitrile (ACN); the mobile phase B is acetonitrile.

[0139] Preferably, the mobile phase A is an aqueous solution of 0.05-0.15% formic acid and 2-7% acetonitrile, and exemplarily an aqueous solution of 0.1% formic acid and 5% acetonitrile.

[0140] Preferably, the quality testing method includes: a purity testing method, a solubility testing method, and a stability testing method.

[0141] Preferably, the high-performance liquid chromatography method employs gradient elution.

[0142] Preferably, the flow rate of the mobile phase is 1 ± 0.2 mL / min; the gradient elution time is 5-60 min; more preferably, it is 10-30 min.

[0143] Preferably, the volume ratio of mobile phase A to mobile phase B in the gradient elution is 1:9-9:1.

[0144] Terminology Definitions and Explanations

[0145] The various terms and phrases used in this invention have their general meanings known to those skilled in the art. Nevertheless, this invention still intends to provide a more detailed description and explanation of these terms and phrases. In the event of any inconsistency between the terms and phrases mentioned and their known meanings, the meanings expressed in this invention shall prevail.

[0146] Unless otherwise stated, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, two or more represent 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. When certain numerical ranges are defined or understood as "numbers," they should be understood to describe the two endpoints of the range, each integer within the range, and each decimal within the range. For example, "numbers from 0 to 10" should be understood to describe not only each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, but also at least the sum of each of these integers with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, and 0.9.

[0147] When this specification and claims refer to a certain value, it includes the value itself, as well as values ​​within a range acceptable in the art before and after that value, such as values ​​within ±15%, ±10%, ±5%, etc. For example, about 10 represents values ​​within: 10 ± 1.5 (i.e., 8.5 to 11.5); 10 ± 1.0 (i.e., 9.0 to 11.0); and 10 ± 0.5 (i.e., 9.5 to 10.5).

[0148] The salts and polymorphs of the compounds of formula I of the present invention can be used in combination with other active ingredients, as long as they do not produce other adverse effects, such as allergic reactions.

[0149] As used in this invention, the term "composition" means a product comprising specified amounts of each specified ingredient, and any product derived directly or indirectly from a combination of specified amounts of each specified ingredient.

[0150] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0151] The term “therapeutic effective dose” refers to the amount of an active compound or drug that researchers, veterinarians, physicians or other clinicians are looking for in a tissue, system, animal, individual or human to elicit a biological or medical response. It includes one or more of the following: (1) prevention of disease: for example, prevention of disease, disorder or condition in an individual who is susceptible to disease, disorder or condition but has not yet experienced or developed the pathology or symptoms of the disease; (2) suppression of disease: for example, suppression of disease, disorder or condition in an individual who is experiencing or developing the pathology or symptoms of the disease (i.e., preventing the further development of the pathology and / or symptoms); (3) relief of disease: for example, relief of disease, disorder or condition in an individual who is experiencing or developing the pathology or symptoms of the disease (i.e., reversal of the pathology and / or symptoms).

[0152] The term "pharmaceutically acceptable" means that a prescription component or active ingredient does not have an excessively harmful effect on health for general therapeutic purposes.

[0153] The term "pharmaceutically acceptable excipient or carrier" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here refers to the ability of the components in the composition to interact with and be mixed with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmacologically acceptable excipients or carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers, wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc. The pharmaceutical compositions may be specifically formulated for oral, parenteral, or rectal administration in solid or liquid form. The pharmaceutical composition can be formulated into various dosage forms to facilitate administration, such as oral formulations (e.g., tablets, capsules, solutions or suspensions) and injectable formulations (e.g., injectable solutions or suspensions, or injectable dry powders that can be used immediately after being added to a drug solvent before injection).

[0154] When used for the above-described therapeutic and / or preventative purposes, the total daily dosage of the salts, polymorphs, and pharmaceutical compositions of Formula I compounds of the present invention must be determined by the attending physician within the bounds of reliable medical judgment. For any given patient, the specific effective therapeutic dose level must be determined based on a number of factors, including the disorder being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors known in the medical field. For example, it is practiced in the art to start the dosage of the compound below the level required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0155] The term "API" refers to a free base, that is, a compound of formula I.

[0156] Beneficial effects

[0157] 1) This invention provides a crystal form of the compound of formula I, which has good pharmaceutical properties. Among them, the free base crystal form A has low hygroscopicity, good solubility and physicochemical stability. The free base crystal form A is thermodynamically stable at room temperature and 50°C, which is beneficial to the storage, quality stability and further drug development of the drug.

[0158] 2) This invention obtained pharmaceutically acceptable salts of compounds of formula I through optimized experimental screening, and further obtained crystal form products of the salts, such as hydrochloride crystal form A, maleate crystal form A, p-toluenesulfonate crystal form A, benzenesulfonate crystal form A, and malonate crystal form A. Among them, p-toluenesulfonate crystal form A has almost no hygroscopicity and good physicochemical stability, higher solubility in biological solvents, and better pharmaceutical value. Attached Figure Description

[0159] Figure 1-1 The XRPD pattern of free alkali crystal form A of compound I;

[0160] Figure 1-2 TGA / DSC plot of free alkali crystal form A of compound I;

[0161] Figure 1-3 For the free alkali crystal form A of compound of formula I 1 H NMR spectrum;

[0162] Figure 1-4 PLM diagram of free alkali crystal form A of compound I;

[0163] Figure 2-1 The XRPD pattern of free alkali crystal form B of compound I is shown.

[0164] Figure 2-2 TGA / DSC plot of free alkali crystal form B of compound I;

[0165] Figure 2-3 For the free alkali crystal form B of compound of formula I 1 H NMR spectrum;

[0166] Figure 2-4 XRPD overlay of free alkali crystal A / B suspension competition test samples;

[0167] Figure 3-1 The XRPD spectrum of the hydrochloride crystal form A of compound I is shown.

[0168] Figure 3-2 For the hydrochloride crystal form A of compound I 1 H NMR spectrum;

[0169] Figure 4-1 The XRPD pattern of maleate crystal form A of compound I is shown.

[0170] Figure 4-2 For maleate crystal form A of compound I 1 H NMR spectrum;

[0171] Figure 5-1 The XRPD pattern of p-toluenesulfonate crystal form A of compound I is shown.

[0172] Figure 5-2 For compound I, p-toluenesulfonate crystal form A 1 H NMR spectrum;

[0173] Figure 6-1 The XRPD spectrum of benzenesulfonate crystal form A of compound I is shown.

[0174] Figure 6-2 TGA / DSC plot of benzenesulfonate crystal form A of compound I;

[0175] Figure 6-3 For the benzenesulfonate crystal form A of compound of formula I 1 H NMR spectrum;

[0176] Figure 7-1 The XRPD pattern of malonate crystal form A of compound I is shown.

[0177] Figure 7-2 TGA / DSC plot of malonate crystal form A of compound I;

[0178] Figure 7-3 For the malonate crystal form A of compound of formula I 1 H NMR spectrum;

[0179] Figure 7-4 The VT-XRPD spectrum of malonate crystal form A of compound I;

[0180] Figure 8-1 XRPD overlay images of samples 1 and 2;

[0181] Figure 8-2 XRPD overlay images of samples 3, 4, 5, and 6;

[0182] Figure 9-1 XRPD pattern of crystal form A of hydrochloride of compound I prepared repeatedly;

[0183] Figure 9-2 TGA / DSC image of crystal form A of hydrochloride of compound I prepared repeatedly;

[0184] Figure 9-3 For the repeated preparation of the hydrochloride crystal form A of compound I. 1 H NMR spectrum;

[0185] Figure 9-4 VT-XRPD pattern of crystal form A of hydrochloride of compound I prepared repeatedly;

[0186] Figure 9-5 PLM diagram of crystal form A of the hydrochloride salt of compound I prepared repeatedly;

[0187] Figure 10-1XRPD pattern of maleate crystal form A of compound I prepared repeatedly;

[0188] Figure 10-2 TGA / DSC plot of maleate crystal form A of compound I prepared repeatedly;

[0189] Figure 10-3 For the repeated preparation of maleate crystal form A of compound I. 1 H NMR spectrum;

[0190] Figure 10-4 VT-XRPD pattern of maleate crystal form A of compound I prepared repeatedly;

[0191] Figure 10-5 PLM diagram of maleate crystal form A of compound I prepared repeatedly;

[0192] Figure 11-1 XRPD pattern of p-toluenesulfonate crystal form A of compound I prepared repeatedly; Figure 11-2 TGA / DSC plot of p-toluenesulfonate crystal form A of compound I prepared repeatedly;

[0193] Figure 11-3 For the repeated preparation of compound I p-toluenesulfonate crystal form A 1 H NMR spectrum;

[0194] Figure 11-4 PLM diagram of p-toluenesulfonate crystal form A of compound I prepared repeatedly;

[0195] Figure 12 This is a dynamic solubility curve at 37℃;

[0196] Figure 13 XRPD overlay image of the solubility of free alkali crystal form A in H2O;

[0197] Figure 14 XRPD overlay image of the solubility of free alkali crystal form A in SGF;

[0198] Figure 15 XRPD overlay of the sample showing the solubility of free alkali crystal form A in FaSSIF;

[0199] Figure 16 XRPD overlay of the sample showing the solubility of free alkali crystal form A in FeSSIF;

[0200] Figure 17 XRPD overlay image of the solubility of hydrochloride crystal form A in H2O;

[0201] Figure 18 XRPD overlay of the sample showing the solubility of hydrochloride crystal form A in SGF;

[0202] Figure 19 XRPD overlay of the sample for the solubility of hydrochloride crystal form A in FaSSIF;

[0203] Figure 20 XRPD overlay of the sample showing the solubility of hydrochloride crystal form A in FeSSIF;

[0204] Figure 21 XRPD overlay image of the solubility of maleate crystal form A in H2O;

[0205] Figure 22 XRPD overlay of the sample showing the solubility of maleate crystal form A in SGF;

[0206] Figure 23 XRPD overlay of the sample for the solubility of maleate crystal form A in FaSSIF;

[0207] Figure 24 XRPD overlay of the sample showing the solubility of maleate crystal form A in FeSSIF;

[0208] Figure 25 XRPD overlay image of the solubility of p-toluenesulfonate crystal form A in H2O;

[0209] Figure 26 XRPD overlay of the sample showing the solubility of p-toluenesulfonate crystal form A in SGF;

[0210] Figure 27 XRPD overlay of the sample for the solubility of p-toluenesulfonate crystal form A in FaSSIF;

[0211] Figure 28 XRPD overlay of the sample for the solubility of p-toluenesulfonate crystal form A in FeSSIF;

[0212] Figure 29 The DVS diagram of free alkali crystal form A;

[0213] Figure 30 XRPD overlay images of free alkali crystal form A before and after DVS testing;

[0214] Figure 31 The DVS diagram of hydrochloride crystal form A;

[0215] Figure 32 XRPD overlay images of hydrochloride crystal form A before and after DVS testing;

[0216] Figure 33 The DVS diagram for maleate crystal form A;

[0217] Figure 34XRPD overlay images of maleate crystal form A before and after DVS test;

[0218] Figure 35 This is the DVS diagram of p-toluenesulfonate crystal form A;

[0219] Figure 36 XRPD overlay images before and after DVS testing of p-toluenesulfonate crystal form A;

[0220] Figure 37 XRPD overlay image of the sample for stability assessment of free alkali crystal form A;

[0221] Figure 38 XRPD overlay image of the sample for evaluating the stability of hydrochloride crystal form A;

[0222] Figure 39 XRPD overlay image of the sample for evaluating the stability of maleate crystal form A;

[0223] Figure 40 XRPD overlay image of the sample for stability assessment of toluenesulfonate crystal form A;

[0224] Figure 41 Results of experiments on the water / water quinine uptake ratio in animals treated with the corresponding compound;

[0225] Figure 42 Results of an experiment on the number of coughs induced by histamine / citric acid stimulation in guinea pigs after administration of the corresponding compounds;

[0226] Figure 43 Results of an experiment on the number of coughs induced by ATP / citric acid stimulation in guinea pigs after administration of the corresponding compound. Detailed Implementation

[0227] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0228] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0229] The instruments and detection methods used in this invention are as follows:

[0230] I. X-ray Powder Diffraction (XRPD)

[0231] XRPD images were acquired using an X-ray powder diffractometer manufactured by PANalytacal, and the scanning parameters are shown in Table 1 below.

[0232] Table 1 XRPD Test Parameters

[0233]

[0234] II. Thermogravimetric analysis (TGA) and Differential scanning calorimetry (DSC)

[0235] The TGA and DSC plots were acquired using a TA 5500 thermogravimetric analyzer and a TA 2500 differential scanning calorimeter, respectively. The test parameters are listed in Table 2 below.

[0236] Table 2 DSC and TGA test parameters

[0237]

[0238] III. Dynamic Moisture Adsorption (DVS)

[0239] Dynamic moisture adsorption (DVS) curves were acquired using the DVSIntrInsic on an SMS (Surface Measurement Systems) device. Relative humidity at 25°C was corrected for the deliquescence points of LiCl, Mg(NO3)2, and KCl. The DVS test parameters are listed in Table 3 below.

[0240] Table 3 DVS Test Parameters

[0241]

[0242] IV. Polarizing Microscope (PLM)

[0243] The polarizing microscope images were taken at room temperature using a Zeiss Axio Scope A1 microscope.

[0244] V. Liquid NMR ( 1 H NMR)

[0245] Liquid NMR spectra were acquired using a Bruker 400M NMR spectrometer with DMSO-d6 as the solvent.

[0246] VI. High Performance Liquid Chromatography (HPLC)

[0247] In the experiment, purity, dynamic solubility, and stability were tested using an Agilent 1260 high-performance liquid chromatograph, and the ion salt molar ratio was tested using ion chromatography. The analytical conditions are shown in Tables 4 and 5 below.

[0248] Table 4. High Performance Liquid Chromatography Test Conditions

[0249]

[0250]

[0251] Table 5 Ion Chromatography Test Conditions

[0252]

[0253] The reagents used in this invention are shown in Table 6 below.

[0254] Table 6. Comparison of Chinese and English names of solvents used in the experiment.

[0255]

[0256] Example 1 Synthesis of Intermediate A-1

[0257]

[0258] Step 1: Synthesis of 3-bromo-2-fluoro-5-iodobenzoic acid

[0259] 3-Bromo-2-fluorobenzoic acid (10 g, 45.7 mmol) was dissolved in concentrated sulfuric acid (40 mL), and NIS (10.27 g, 45.7 mmol) was added in portions at 0 °C. The mixture was stirred at room temperature for three hours. The solution was quenched with ice water (200 mL), filtered, and the filter cake was washed five times with water (200 mL) and dried under vacuum to obtain 3-bromo-2-fluoro-5-iodobenzoic acid (10.9 g, white solid, yield 69.2%).

[0260] Step 2: Synthesis of 3-bromo-2-fluoro-5-hydroxybenzoic acid

[0261] Cuprous oxide (0.656 g, 4.74 mmol) was added to a solution of 3-bromo-2-fluoro-5-iodobenzoic acid (10.9 g, 31.6 mmol) and sodium hydroxide (6.32 g, 158 mmol) in water (100 mL), and the mixture was reacted overnight at 100 °C. After cooling to room temperature, the mixture was filtered, and the pH of the filtrate was adjusted to 1 with 2 M hydrochloric acid solution. The filtrate was extracted with ethyl acetate (60 mL × 3), and the organic phase was concentrated to dryness to give 3-bromo-2-fluoro-5-hydroxybenzoic acid (7.2 g, yellow solid, yield 96.8%).

[0262] Step 3: Synthesis of methyl 3-bromo-2-fluoro-5-hydroxybenzoate

[0263] A methanol (120 mL) solution of 3-bromo-2-fluoro-5-hydroxybenzoic acid (7.2 g, 30.6 mmol) was added with thionyl chloride (10.9 g, 91.8 mmol), and the mixture was stirred at 55 °C for 16 hours. The solvent was then removed under reduced pressure, and the solution was concentrated to give a solid compound, methyl 3-bromo-2-fluoro-5-hydroxybenzoate (3.1 g, yield 40.8%), which was used in the next step without further purification.

[0264] Step 4: Synthesis of methyl 2-fluoro-5-hydroxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate

[0265] Methyl 3-bromo-2-fluoro-5-hydroxybenzoate (3.1 g, 12.45 mmol), pinacol diboronate (3.48 g, 13.69 mmol), and potassium acetate (3.67 g, 37.3 mmol) were dissolved in 1,4-dioxane (50 mL), and the solution was degassed with a stream of nitrogen for 2 minutes. Pd(dppf)Cl2 (0.455 g, 0.622 mmol) was added, and the resulting solution was degassed with a stream of nitrogen for another 2 minutes. The reaction mixture was then stirred at 100 °C for 16 hours. The reaction mixture was filtered and concentrated under vacuum. The residue was purified by silica gel column chromatography to give methyl 2-fluoro-5-hydroxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate (3.4 g, white solid, 92% yield).

[0266] Step 5: Synthesis of methyl 2-fluoro-5-hydroxy-3-(5-methylthiazolyl-2-yl)benzoate (intermediate A-1)

[0267] At room temperature, Pd(dppf)Cl2 (1.260 g, 1.722 mmol) was added to a mixed solution of methyl 2-fluoro-5-hydroxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate (3.4 g, 11.48 mmol), 2-bromo-5-methylthiazole (2.453 g, 13.78 mmol), potassium carbonate (3.81 g, 27.6 mmol) in THF (30 mL) and water (10 mL). After three purgings under vacuum with nitrogen, the reaction was carried out at 90 °C for 16 h. Dilute with water (30 mL), extract with ethyl acetate (40 mL × 3), concentrate the organic phase to dryness, and separate and purify the residue by silica gel column chromatography to obtain methyl 2-fluoro-5-hydroxy-3-(5-methylthiazolyl-2-yl)benzoate (intermediate A-1, 1.41 g, yellow solid, yield 45.9%).

[0268] LC-MS, M / Z: 268.2 [M+H] + .

[0269] Example 2 Preparation of Compound I

[0270]

[0271] Step 1: Synthesis of (4R,5R)-4,5-dimethyl-1,3,2-dioxathiacyclopentane-2-oxide

[0272] (2R,3R)-(-)-2,3-butanediol (2 g, 22.19 mmol) and pyridine (3.86 g, 48.8 mmol) were dissolved in dry tetrahydrofuran (20 mL). The reaction temperature was adjusted to 0-5 °C, and thionyl chloride (2.9 g, 24.41 mmol) was slowly added. The mixture was heated to room temperature and stirred for 16 h. The reaction was quenched with water (30 mL), and 20 mL of ethyl acetate was added. The mixture was separated, and the organic phase was washed with saturated ammonium chloride (20 mL) and saturated sodium chloride aqueous solution (20 mL). The solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain (4R,5R)-4,5-dimethyl-1,3,2-dioxathiacyclopentane-2-oxide (2.4 g, colorless liquid, yield 79%).

[0273] 1 H NMR (400MHz, Chloroform-d) δ4.63 (dq, J=9.0, 6.1Hz, 1H), 4.07 (dq, J=9.0, 6.1Hz, 1H), 1.52 (d, J=6.2Hz, 3H), 1.43 (d, J=6.1Hz, 3H).

[0274] Step 2: Synthesis of methyl 2-fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazolyl-2-yl)benzoate

[0275] Under nitrogen protection, cesium carbonate (1.22 g, 3.74 mmol) was added to a solution of methyl 2-fluoro-5-hydroxy-3-(5-methylthiazolyl-2-yl)benzoate (intermediate A-1, 500 mg, 1.871 mmol) in N,N-dimethylformamide (5 mL), and the mixture was stirred at room temperature for 30 min. Then, (4R,5R)-4,5-dimethyl-1,3,2-dioxathiacyclopentane-2-oxide (382 mg, 2.81 mmol) was added, and the mixture was heated to 80 °C and reacted for 16 h, followed by cooling to room temperature. The reaction solution was concentrated to dryness under reduced pressure, and chloroform (20 mL) and 4M sulfuric acid solution (20 mL) were added. The mixture was stirred at 70 °C for 5 h, separated, and the pH of the aqueous phase was adjusted to 7-8 with sodium bicarbonate. It was extracted with dichloromethane (20 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 2:1) to give methyl 2-fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazolyl-2-yl)benzoate (380 mg, white solid, yield 60%).

[0276] LC-MS, M / Z: 340.1 [M+H] + .

[0277] Step 3: Synthesis of 2-fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazol-2-yl)benzoic acid

[0278] Methyl 2-fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazol-2-yl)benzoate (200 mg, 0.589 mmol) was dissolved in methanol (4 mL), followed by the addition of lithium hydroxide monohydrate (70.7 mg, 1.765 mmol) and water (0.4 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction solution was concentrated to dryness, and water (5 mL) was added. The pH was adjusted to 2-3 with 1 M hydrochloric acid solution. The aqueous phase was extracted with dichloromethane (5 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain 2-fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazol-2-yl)benzoic acid (190 mg, white solid, 99% yield).

[0279] LC-MS, M / Z: 326.1 [M+H] + .

[0280] Step 4: 2-Fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazolyl-2-yl)-N-((R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide

[0281]

[0282] Under nitrogen protection, 2-fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazolyl-2-yl)benzoic acid (190 mg, 0.584 mmol), (R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl-1-amine hydrochloride (160 mg, 0.701 mmol), N,N-diisopropylethylamine (226 mg, 1.752 mmol), and N,N-dimethylformamide (5 mL) were added sequentially to the reaction flask. The mixture was cooled to approximately 0°C, and a 50% solution of 1-propylphosphonic anhydride in N,N-dimethylformamide (557 mg, 0.876 mL) was added dropwise. After adding ol), the mixture was allowed to return to room temperature for 16 hours. The reaction was then quenched with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL × 3), and the organic phases were combined. The organic phases were washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, concentrated, and the residue was separated and purified by silica gel (petroleum ether: ethyl acetate (V / V) = 1:1) to give a white solid 2-fluoro-5-(((2S,3R)-3-hydroxybut-2-yl)oxy)-3-(5-methylthiazolyl-2-yl)-N-((R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (compound of formula I, 110 mg, yield 37.8%).

[0283] 1 H NMR (400MHz, CDCl3) δ8.94 (s, 2H), 7.86 (dd, J = 5.9, 3.3Hz, 1H), 7.63–7.58 (m, 1H), 7.50 (dd,J=5.8,3.4Hz,1H),7.10(dd,J=12.3,6.5Hz,1H),5.38(ddd,J=7.7,4.4,1.5Hz,1H), 4.40(qd,J=6.3,3.3Hz,1H),4.01(ddd,J=6.5,4.8,3.3Hz,1H),2.56(d,J=1.2Hz,3H),2. 07(d,J=4.9Hz,1H),1.72(d,J=7.1Hz,3H),1.26(d,J=6.3Hz,3H),1.23(d,J=6.5Hz,3H).

[0284] LC-MS, M / Z: 499.1 [M+H] + .

[0285] Example 3: Preparation of free base crystal form A of compound I

[0286] Weigh approximately 20 mg of compound I into an HPLC vial, add 0.5 mL of toluene, and stir the resulting suspension magnetically (1000 rpm) at room temperature for approximately 4 days. Then, centrifuge (10000 rpm, 2 min) to collect the solid and perform XRPD analysis. Figure 1-1 As shown, based on the test results, the solid product is designated as free alkali crystal form A. TGA / DSC results ( Figure 1-2 The data shows a 1.28% weight loss when the sample is heated to 150℃, and an endothermic peak at 175.6℃ (initial temperature). Free alkali crystal form A... 1 The H NMR results are as follows Figure 1-3 As shown. PLM( Figure 1-4 The sample appears as aggregated needle-like crystals.

[0287] The XRPD analytical data of the free basal crystal form A of the obtained compound of formula I are shown in Table 7 below:

[0288] Table 7 XRPD diffraction peak data of free alkali crystal form A

[0289]

[0290]

[0291] Example 4: Preparation of free acid crystal form B of compound I

[0292] The free alkali crystal form A of compound I was dissolved in 1,4-dioxane and then subjected to gas-liquid diffusion in an n-Hexane atmosphere. The solid was then dried at room temperature to obtain the free alkali crystal form B.

[0293] XRPD of free alkali crystal form B, such as Figure 2-1 As shown. TGA / DSC results ( Figure 2-2 The data shows a 2.36% weight loss when the sample is heated to 150℃, and an endothermic peak at 177.0℃ (initial temperature). Free alkali crystal form B... 1 The H NMR results are as follows Figure 2-3 As shown.

[0294] The XRPD analytical data of the free basal crystal form B of the obtained compound of formula I are shown in Table 8 below:

[0295] Table 8 XRPD diffraction peak data of free alkali crystal form B

[0296]

[0297]

[0298] To confirm the thermodynamic transformation relationship between free alkali crystal forms A and B under different temperature conditions, a suspension competition experiment was conducted in n-heptane and toluene at room temperature and 50°C. The specific steps are as follows: 1) First, prepare saturated solutions of the initial sample, free alkali crystal form A, at the corresponding temperature and solvent; 2) Weigh an appropriate amount of the mixed crystal sample and add it to the filtered saturated solution to form a suspension; 3) Suspend and stir the sample at the corresponding temperature conditions. The results are summarized in Table 9, and the XRPD results are summarized in [Table data missing]. Figure 2-4 The results showed that after suspension competition at 50℃ in the n-heptane system and at room temperature / 50℃ in the toluene system, both alkali crystals converted to free alkali form A. In the n-heptane system, after 42 days of suspension competition at room temperature, free alkali crystal form A was also converted. The experiment indicates that free alkali crystal form A is thermodynamically stable at both room temperature and 50℃.

[0299] Table 9 Summary of Hybrid Suspension Competition Test

[0300]

[0301] Example 5: Preparation and Screening of Salt Forms of Compound I

[0302] Approximately 20 mg of compound I and equimolar amounts of different salt-forming agents (i.e., acids that form salts with free bases) were weighed into an HPLC vial. 0.5 mL of solvent was added to mix and obtain a suspension. The salt-forming agents were first diluted with their respective solvents before being mixed with the starting sample. After suspending and stirring at room temperature for approximately 5 days, the solid was centrifuged and vacuum-dried overnight at room temperature. For the room-temperature clear system, 0.5–1.0 mL of the antisolvent n-heptane was added to increase the supersaturation of the solution and accelerate crystallization. XRPD characterization of the obtained solids showed that five salt types were obtained in the salt type screening test (Table 10).

[0303] Table 10 Summary of Salt Type Screening Test Results

[0304]

[0305]

[0306] # The acid / alkali molar ratio is 1:1.

[0307] *: After stirring at room temperature for 3 hours to obtain a clear solution, 0.5-1.0 mL of n-heptane was added and solid precipitation was observed.

[0308] Example 6: Preparation of the hydrochloride crystal form A of compound I

[0309] After mixing the compound sample of Formula I with an equimolar amount of hydrochloric acid in MTBE at room temperature for 5 days, the solid was separated by centrifugation and then vacuum dried at room temperature to obtain hydrochloric acid crystal form A.

[0310] The XRPD pattern of sample A of hydrochloride crystal form is shown below. Figure 3-1 As shown. 1 H NMR was measured in DMSO-d6, and the results are as follows: Figure 3-2 The results showed that no MTBE solvent residue was observed in the sample. HPLC / IC results indicated that the molar ratio of hydrochloric acid to API (i.e., free base) was 0.5:1.

[0311] The XRPD analytical data of the hydrochloride crystal form A of the obtained compound of formula I are shown in Table 11 below:

[0312] Table 11 XRPD diffraction peak data of hydrochloride crystal form A

[0313]

[0314] Example 7 Preparation of maleate crystal form A of compound I

[0315] The sample of compound I and an equimolar amount of maleic acid were stirred in EtOAc at room temperature for 3 hours. Then, 0.5 mL of n-heptane was added to the resulting clear solution and stirred at room temperature for 5 days. The solid was then separated by centrifugation and vacuum dried at room temperature to obtain the maleate crystal form A of compound I.

[0316] The XRPD image of sample A, maleate crystal form, is shown below. Figure 4-1 As shown. 1 H NMR was measured in DMSO-d6, and the results are shown below. Figure 4-2 The results showed that the molar ratio of maleic acid to API (i.e., free base) was 0.5:1, and no solvent residues of EtOAc and n-heptane were observed.

[0317] The XRPD analytical data of the maleate crystal form A of the obtained compound of formula I are shown in Table 12 below:

[0318] Table 12 XRPD diffraction peak data of maleate crystal form A

[0319]

[0320] Example 8: Preparation of p-toluenesulfonate crystal form A of compound I

[0321] The sample of compound I and an equimolar amount of p-toluenesulfonic acid were stirred in EtOAc at room temperature for 5 days. After centrifugation, the solid was separated and dried under vacuum at room temperature to obtain the p-toluenesulfonate crystal form A of compound I.

[0322] The XRPD pattern of sample A of p-toluenesulfonate crystal form is shown below. Figure 5-1 As shown. 1 H NMR was measured in DMSO-d6, and the results are shown below. Figure 5-2 The results showed that the molar ratio of p-toluenesulfonic acid to API (i.e., free base) was 1.0:1, and no EtOAc solvent residue was observed.

[0323] The XRPD analysis data of the p-toluenesulfonate crystal form A of the obtained compound of formula I are shown in Table 13 below:

[0324] Table 13 XRPD diffraction peak data of p-toluenesulfonate crystal form A

[0325]

[0326] Example 9 Preparation of benzenesulfonate crystal form A of compound I

[0327] After stirring the sample of compound I and an equimolar amount of benzenesulfonic acid in MTBE at room temperature for 5 days, the solid was separated by centrifugation and dried under vacuum at room temperature to obtain the benzenesulfonate crystal form A of compound I.

[0328] The XRPD diagram of benzenesulfonate crystal form A is shown below. Figure 6-1 As shown. See details of the TGA / DSC results. Figure 6-2 TGA results showed that the sample lost 1.35% of its weight when heated to 120℃; DSC results showed that the sample had one endothermic peak, with an initial temperature of 159.6℃ and a peak temperature of 160.9℃. 1 H NMR was measured in DMSO-d6, and the results are shown below. Figure 6-3 The results showed that the molar ratio of benzenesulfonic acid to API (i.e., free base) was 0.9:1, and no solvent residue of MTBE was observed.

[0329] The XRPD analysis data of the benzenesulfonate crystal form A of the obtained Formula I compound are shown in Table 14 below:

[0330] Table 14 XRPD diffraction peak data of benzenesulfonate crystal form A

[0331]

[0332]

[0333] Example 10: Preparation of malonate crystal form A of compound I

[0334] Compound of Formula I and an equimolar amount of malonic acid were stirred in EtOAc at room temperature for ~3 hours. Then, 0.5 mL of n-heptane was added to the resulting clear liquid and stirred at room temperature for 5 days. After centrifugation, the solid was separated and dried under vacuum at room temperature to obtain the malonate crystal form A of compound of Formula I.

[0335] The XRPD pattern of malonate crystal form A is shown below. Figure 7-1 As shown. See details of the TGA / DSC results. Figure 7-2 TGA results showed that the sample lost 2.99% of its weight when heated to 120℃ and 11.02% of its weight when heated from 120℃ to 200℃; DSC results showed that the sample had an endothermic peak at 156.4℃ (peak temperature). 1 H NMR was measured in DMSO-d6, and the results are shown below. Figure 7-3 The results showed that the molar ratio of malonic acid to API (i.e., free base) was 0.7:1, and no solvent residue of EtOAc was observed.

[0336] via VT-XRPD ( Figure 7-4 The crystal form of malonate A was identified. The results showed that no crystal form change was observed when the sample was purged under N2 for 20 min, heated to 120℃ and cooled to room temperature.

[0337] The XRPD analytical data of the malonate crystal form A of the obtained compound of formula I are shown in Table 15 below:

[0338] Table 15 XRPD diffraction peak data of malonate crystal form A

[0339]

[0340]

[0341] Example 11 Repeated preparation of polymorphs of Formula I compound (I) Repeated preparation of free base crystal form B

[0342] Table 16 Summary of Repeated Preparation Results of Free Alkali Crystal Form B

[0343]

[0344] As shown in Table 16, free alkali crystal form A was not converted into free alkali crystal form B in TFE; it remained free alkali crystal form A (XRPD patterns of samples 1 and 2 are shown in Table 16). Figure 8-1 (As shown); mixed crystals of free alkali crystal form A+B were obtained in 1,4-dioxane (XRPD patterns of samples 3, 4, 5, and 6 are shown in the figure). Figure 8-2 (As shown).

[0345] (II) Repeated preparation of salt forms

[0346] Three salt forms, namely hydrochloride form A, maleate form A, and p-toluenesulfonate form A, were prepared in 300 mg repeated batches. The results showed that all three salt forms were successfully prepared repeatedly. The repeated preparation steps for the salt forms are summarized in Table 17.

[0347] Table 17 Summary of Salt Form Repeated Preparation Steps

[0348]

[0349]

[0350] 1. Hydrochloride crystal form A

[0351] The XRPD pattern of the repeatedly prepared hydrochloride crystal form A sample is shown below. Figure 9-1 As shown. The TGA / DSC results are shown below. Figure 9-2 TGA results showed that the sample lost 2.2% of its weight when heated to 100℃ and 3.9% of its weight when heated from 100℃ to 160℃. DSC results showed that the sample had endothermic peaks at 157.4℃ and 179.0℃ (peak temperature). 1 H NMR was measured in DMSO-d6, and the results are as follows: Figure 9-3 As shown, no MTBE solvent residue was observed. HPLC / IC results indicate that the molar ratio of hydrochloride crystal form A sample is 0.5:1 (hydrochloric acid:API). PLM( Figure 9-5 The results showed that hydrochloride crystal form A consisted of irregular small particles. This was confirmed by VT-XRPD (…). Figure 9-4 The crystal form of hydrochloride A was identified. The results showed that no crystal form change was observed when the sample was purged under N2 for 20 min, heated to 100℃ and cooled to room temperature.

[0352] 2. Maleate crystal form A

[0353] The XRPD pattern of the repeatedly prepared maleate crystal form A sample is shown below. Figure 10-1 As shown. The TGA / DSC results are shown below. Figure 10-2 TGA results showed that the sample lost 1.65% of its weight when heated to 110℃, and 11.88% of its weight when heated from 110℃ to 220℃; DSC results showed that the sample had an endothermic peak at 144.1℃ (peak temperature). 1 H NMR was measured in DMSO-d6, and the results are shown below. Figure 10-3 The results showed that the molar ratio of maleic acid to API in the sample was 0.5:1, and 0.3 wt% of EtOAc solvent residue was observed. PLM( Figure 10-5 The results showed that maleate crystal form A consisted of irregular particles with some agglomeration. This was confirmed by VT-XRPD (…). Figure 10-4The crystal form of maleate A was identified. The results showed that no crystal form change was observed when the sample was purged under N2 for 20 min, heated to 120℃ and cooled to room temperature.

[0354] 3. p-Toluenesulfonate crystal form A

[0355] The XRPD pattern of the repeatedly prepared p-toluenesulfonate crystal form A sample is shown below. Figure 11-1 As shown. The TGA / DSC results are shown below. Figure 11-2 TGA results showed that the sample lost 0.73% of its weight when heated to 150℃; DSC results showed that the sample had one endothermic peak, with an initial temperature of 157.1℃ and a peak temperature of 159.2℃. 1 H NMR was measured in DMSO-d6, and the results are shown below. Figure 11-3 The results showed that the molar ratio of p-toluenesulfonic acid to API in the sample was 1.0:1, and no EtOAc solvent residue was observed. PLM( Figure 11-4 The results show that p-toluenesulfonate crystal form A is in the form of irregularly shaped particles.

[0356] Example 12 Dynamic Solubility Experiment

[0357] Solid feed concentration of 10 mg / mL (based on free alkali) was used for rotary mixing at 37 °C, and the concentrations of each sample in water, SGF, FaSSIF, and FeSSIF were determined at different time points (1, 4, and 24 hours). 1 Solubility in four systems. Samples were taken at each time point, centrifuged (10000 rpm), and filtered (using a 0.45 μm PTFE filter). The HPLC concentration and pH of the filtrate were measured. The solubility test results are summarized in Table 12-1, and the solubility curves are shown below. Figure 12 The results showed that maleate crystal form A had the highest solubility in H2O, while p-toluenesulfonate crystal form A had the highest solubility in three biological solvents other than H2O within 1 hour. Figure 13-28 XRPD overlay images of the solubility samples of free alkali crystal form A, hydrochloride crystal form A, maleate crystal form A, and p-toluenesulfonate crystal form A in H2O, SGF, FaSSIF, and FeSSIF, respectively. The crystal form changes are shown in Table 18. The crystal form of free alkali crystal form A did not change during the dynamic solubility test, showing high stability.

[0358] Table 18 Summary of Dynamic Solubility Test Results at 37℃

[0359]

[0360]

[0361] S: Solubility (mg / mL); a: Transforms into free alkali crystal form A; b : Transforms into free alkali crystal form B;

[0362] a ':Transforms into a free alkali crystal form A+ additional diffraction peak; b ':Transforms into an additional diffraction peak of free alkali crystal B+;

[0363] a+b ':Transformed into free alkali crystal form A+ free alkali crystal form B+ additional diffraction peaks.

[0364] Instructions for preparing biological solvents:

[0365] Preparation of simulated gastric juice (SGF):

[0366] Weigh 100.2 mg NaCl and 50.3 mg Trinaton X-100 into a 50 mL volumetric flask and dissolve in purified water. Add 816 μL of 1M hydrochloric acid and adjust the pH to 1.8 with 1M hydrochloric acid or 1M NaOH solution. Make up to volume with purified water.

[0367] Preparation of intestinal fluid simulating fasting (FaSSIF):

[0368] Weigh 170.5 mg of anhydrous NaH₂PO₄, 22.2 mg of NaOH, and 310.9 mg of NaCl into a 50-mL volumetric flask. Dissolve in purified water and adjust the pH to 6.5 with 1M hydrochloric acid or 1M NaOH solution. Make up to volume with purified water. Then weigh 55.1 mg of SIF powder into a 25-mL volumetric flask, dissolve in the above solution, and make up to volume.

[0369] Preparation of intestinal fluid under simulated feeding conditions (FeSSIF):

[0370] Take 0.21 mL of glacial acetic acid, 101.6 mg of NaOH, and 295.4 mg of NaCl into a 25 mL volumetric flask. Add approximately 20 mL of purified water to dissolve, and adjust the pH to 5.0 with 1 M hydrochloric acid or 1 M NaOH solution. Make up to volume with purified water, and weigh in 280.8 mg of SIF powder to dissolve.

[0371] Example 13 Hygroscopicity

[0372] The hygroscopicity of the free alkali crystal form A prepared in Example 3, and the hydrochloride crystal form A, maleate crystal form A, and p-toluenesulfonate crystal form A prepared repeatedly in Example 11, was evaluated using a dynamic moisture adsorption (DVS) instrument. Starting at 0% RH, the percentage change in sample mass was collected under a constant temperature of 25°C as humidity changed (0% RH → 95% RH → 0% RH). The DVS test results and the XRPD results of the samples before and after the DVS test are shown below. Figures 29-36As shown in the figure. The results show that the free alkali crystal form A has a moisture adsorption of ~0.91% at 25℃ / 80%RH, indicating that it is slightly hygroscopic; the hydrochloride crystal form A has a moisture adsorption of ~0.49% at 25℃ / 80%RH, indicating that it is slightly hygroscopic; the maleate crystal form A has a moisture adsorption of ~0.41% at 25℃ / 80%RH, indicating that it is slightly hygroscopic, and rapid moisture absorption was observed in maleate crystal form A when the humidity is >80%RH; the p-toluenesulfonate crystal form A sample has a moisture adsorption of ~0.12% at 25℃ / 80%RH, indicating that it is almost non-hygroscopic. The crystal forms of the free alkali crystal form A, hydrochloride crystal form A, maleate crystal form A, and p-toluenesulfonate crystal form A samples remained unchanged after DVS testing.

[0373] Example 14 Solid-state stability

[0374] The free alkali crystal form A prepared in Example 3, and the hydrochloride crystal form A, maleate crystal form A, and p-toluenesulfonate crystal form A prepared repeatedly in Example 11 were placed in a closed container at 60°C for 1 day, in an open container at 25°C / 60%RH for 1 week, and in an open container at 40°C / 75%RH for 1 month (hydrochloride crystal form A and maleate crystal form A were only placed for 1 week). The physical and chemical stability of the samples was determined by XRPD and HPLC. Purity data are listed in Table 19, and XRPD results are listed in... Figures 37-40 The results showed that the HPLC purity of free alkali crystal form A, hydrochloride crystal form A, maleate crystal form A, and p-toluenesulfonate crystal form A did not change significantly after being placed under the corresponding conditions.

[0375] Table 19 Summary of Solid State Stability Assessment

[0376]

[0377]

[0378] Example 15: Different preparation processes of the free alkali crystal form of compound I

[0379] Compound of Formula I was prepared according to the process method of Example 2, and free alkali crystal form A or free alkali crystal form B (identified by XRPD) can be prepared according to the following method.

[0380] Method 1: Slow evaporation

[0381] Six slow evaporation tests were conducted using different solvent systems. Approximately 20 mg of each Formula I compound sample was weighed into a 5 mL vial, and 0.4–0.6 mL of the solvents listed in Table 15-1 was added. After dissolution, the vials were filtered, sealed with sealing film, and two pinholes were punched in the film. The vials were then left to evaporate slowly at room temperature. The resulting solids were collected and subjected to XRPD testing. The experimental results are shown in Table 20, yielding free alkali crystal forms A / B.

[0382] Table 20 Summary of Slow Evaporation Test

[0383]

[0384] Method 2: Gas-solid permeation

[0385] Eight gas-solid permeation tests were conducted using different solvents. Approximately 20 mg of each Formula I compound sample was weighed into a 3 mL vial, and approximately 3 mL of solvent was added to a 20 mL vial. The 3 mL vial was placed open inside the 20 mL vial, which was then sealed. After standing at room temperature for 1–8 days, the solids were collected and subjected to XRPD testing. The results are shown in Table 21, yielding free alkali crystalline form A and free alkali crystalline form A+B.

[0386] Table 21 Summary of Gas-Solid Diffusion Experiments

[0387]

[0388]

[0389] *: Obtained by evaporation at room temperature after clarification.

[0390] Method 3: Gas-liquid osmosis

[0391] Eight gas-liquid permeation tests were conducted using different solvents. Approximately 20 mg of each Formula I compound sample was weighed into a 3 mL vial, and dissolved in 0.4–2.2 mL of solvent (filtered using a 0.45 μm PTFE filter). Separately, approximately 3 mL of the antisolvent was added to a 20 mL vial. The 3 mL vial containing the supernatant was placed open over the 20 mL vial, and the 20 mL vial was sealed and allowed to stand at room temperature. The resulting solids were collected and subjected to XRPD testing. The results are shown in Table 22, revealing free alkali crystal forms A / B and low crystallinity.

[0392] Table 22 Summary of Gas-Liquid Diffusion Experiments

[0393]

[0394] *: Obtained by volatilization at room temperature.

[0395] Method 4: Polymer Induction

[0396] Eight polymer-induced experiments were conducted using two mixed polymers in different solvents. Approximately 20 mg of each Formula I compound sample was weighed into a 3 mL vial, dissolved in 0.4–1.0 mL of solvent, filtered, and then approximately 2 mg of the mixed polymer was added. The vial was sealed with sealing film, with two small holes punched in the film, and left to evaporate slowly at room temperature. The experimental results are shown in Table 23, yielding free alkali crystalline form A and free alkali crystalline form A+B.

[0397] Table 23 Summary of Polymer Induction Tests

[0398]

[0399] Mixed polymer A: Polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose, and methylcellulose (mixed in equal masses)

[0400] Mixed polymer B: polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate, and hydroxyethyl cellulose (mixed in equal masses).

[0401] Method 5: Humidity Induction

[0402] Five humidity-induced experiments were conducted using different solvents. Approximately 20 mg of each sample of compound I was weighed into a 3 mL vial. A saturated salt solution at room temperature was prepared in a separate 20 mL vial. The 3 mL vial was placed open inside the 20 mL vial, which was then sealed. After standing at room temperature for 8 days, the solid was collected and subjected to XRPD testing. The experimental results are shown in Table 24, yielding the free alkali crystalline form A.

[0403] Table 24 Summary of Humidity Induction Test

[0404]

[0405] Method 6: Circulating heating and cooling

[0406] Nine cycles of heating and cooling were conducted using different solvents. Approximately 20 mg of each Formula I compound sample was weighed into an HPLC glass vial, and 0.5 mL of each of the solvents listed in Table 15-6 was added. The resulting suspension was subjected to temperature cycling (50℃~5℃, 0.1℃ / min, 2 cycles) with magnetic stirring (1000 rpm), centrifuged (10000 rpm, 2 min), and the solid was collected for XRPD analysis. The experimental results are shown in Table 25, yielding free alkali crystal form A.

[0407] Table 25 Summary of Cyclic Heating and Cooling Tests

[0408]

[0409] Method 7: Adding antisolvent

[0410] Twelve antisolvent addition experiments were conducted using different solvents. Approximately 20 mg of each Formula I compound sample was weighed and added to a 20 mL vial. The solid was completely dissolved using 0.4–0.6 mL of solvent (see Table 15-7). The antisolvent from Table 15-7 was added dropwise to the clear solution while stirring (1000 rpm) until solid precipitated, or until the total volume of antisolvent reached 5 mL. Samples without solid precipitation were then suspended and stirred at 5°C. If no solid precipitate was observed, the samples were suspended and stirred at -20°C. The final clear sample was allowed to evaporate at room temperature. The precipitated solid was separated and XRPD was performed. The results are shown in Table 26. The antisolvent addition experiments yielded free alkali crystal form A and a clear solution.

[0411] Table 26 Summary of Antisolvent Addition Experiments

[0412]

[0413] *: After adding the antisolvent, a clear liquid was obtained, which was then stirred at 5°C.

[0414] Method 8: Addition of anti-antisolvents

[0415] Eight anti-antisolvent addition experiments were conducted using different solvents. Approximately 20 mg of each Formula I compound sample was weighed and added to a 20 mL vial. The solid was completely dissolved in 0.4–0.6 mL of solvent (see Table 15-8). This clear solution was then added dropwise to 5 mL of antisolvent (as shown in Table 15-8) while stirring (1000 rpm). Samples without solid precipitation were suspended and stirred at 5 °C. If no solid still precipitated, the samples were suspended and stirred at -20 °C. The final clear sample was allowed to evaporate at room temperature. The precipitated solid was separated and XRPD was performed. The results are shown in Table 27. The antisolvent addition experiment yielded free alkali crystal form A.

[0416] Table 27 Summary of Anti-Antisolvent Addition Experiments

[0417]

[0418] *: The sample was obtained by stirring at 5°C; # The sample was obtained by stirring at -20℃.

[0419] Method 9: Room temperature suspension stirring

[0420] Fifteen room-temperature suspension-stirring experiments were conducted using different solvents. Approximately 20–40 mg of each Formula I compound sample was weighed into HPLC glass vials, and 0.5 mL of each solvent listed in Table 15-9 was added. The resulting turbid solution was magnetically stirred at room temperature (1000 rpm) for approximately 4 days, then centrifuged (10000 rpm, 2 min) to collect the solid and perform XRPD analysis. The experimental results are shown in Table 28, yielding free alkali crystal form A.

[0421] Table 28 Summary of Room Temperature Suspension Stirring Test

[0422]

[0423]

[0424] Method 10: Slowly lower the temperature

[0425] Five slow cooling experiments were conducted using different solvent systems. Approximately 20 mg of each Formula I compound sample was weighed into an HPLC vial, and 1.0 mL of the solvent listed in Table 15-10 was added. After stirring and equilibration at 50°C for approximately 2 hours, the sample was filtered (using a 0.45 μm PTFE filter) to collect the supernatant. The supernatant was placed in a biological incubator and cooled from 50°C to 5°C at a rate of 0.05°C / min, then maintained at 5°C. The clear solution was then transferred to -20°C for isothermal control. The precipitated solid was collected and subjected to XRPD testing. Samples without precipitated solids were transferred to room temperature for evaporation. The experimental results are shown in Table 29. The slow cooling experiments yielded multiple peaks for free basal crystal form A and free basal crystal form B+.

[0426] Table 29 Summary of the Slow Cooling Experiment

[0427]

[0428] *: The sample was obtained by volatilization at room temperature.

[0429] Method 11: Suspension stirring at 50℃

[0430] Sixteen suspension-stirring experiments were conducted at 50°C using different solvents. Approximately 20–40 mg of each Formula I compound sample was weighed into HPLC glass vials, and 0.5 mL of each solvent listed in Table 15-11 was added. The resulting suspensions were magnetically stirred at 50°C (1000 rpm) for approximately 3 days, then centrifuged (10000 rpm, 2 min) to collect the solids and perform XRPD analysis. The experimental results are shown in Table 30, revealing the free alkali crystal form A.

[0431] Table 30 Summary of the 50℃ Suspension Stirring Test

[0432]

[0433]

[0434] *: The sample was clarified at 50°C and then stirred at room temperature.

[0435] Example 16

[0436] Control compound 1 and control compound 2 were synthesized with reference to patent application WO 2016 / 091776A1.

[0437]

[0438] I. Determination of the antagonistic activity of hP2X3 antagonists against hP2X3 by FLIPR method

[0439] The antagonistic activity of human P2X3 receptor (hP2X3) antagonists against hP2X3 was evaluated using the FLIPR Calcium4Assay Kit (Molecular Devices, R8141) and the FLIPR TETRA instrument (MolecularDevices, 0296) to detect calcium flow signals. Twenty-four hours before the experiment, human cells stably transfected with the hP2X3 receptor were cultured at a rate of 2 × 10⁻⁶ cells / year. 5 Cells were seeded at a density of 50 μL / mL in 384-well plates and incubated at 37°C for 16–24 h in a 5% CO2 incubator. 500 nmol of the desired concentration of the test compound was prepared using DMSO stock solution (20–50 mM DMSO), and added to each well of the 384-well plate. 30 μL of FLIPR Assay buffer (containing 1.26 mM Ca) was then added to the top of each well. 2+ Mix 1×HBSS + 2mM CaCl2 + 20mM HEPES, shake for 20-40 min to mix. Prepare 3 times the required concentration of agonist (α,β-meATP) using FLIPR Assay buffer (final concentration required 400 nM), and add 45 μL of agonist to each well of another 384-well plate. Take the cell culture plate from the previous day, aspirate the cell supernatant, and add 30 μL of Dye to each well. The cells were incubated for 1 hour using a Calcium 4 Assay Kit (diluted with FLIPR buffer). 15 μL of the compound was added to each well (using a FLIPR instrument). After 15 minutes, 22.5 μL of the agonist was added to each well, and the fluorescence signal was detected (excitation wavelength 470 nm-495 nm, emission wavelength 515 nm-575 nm). The difference between the peak and trough values ​​was used as the baseline data. The highest concentration of the positive control was taken as the 100% inhibition rate, and DMSO data as the 0% inhibition rate. The inhibition effect curve of the compound was fitted using GraphpadPrism 6 software, and the IC50 was calculated. 50 value.

[0440] Table 31 shows the antagonistic activity of the tested compounds against hP2X3.

[0441] Test compounds <![CDATA[hP2X3 IC 50 (nM)]]> Reference compound 1 241 Formula I compound 67

[0442] II. Mouse Pharmacokinetic Study

[0443] Pharmacokinetic studies were conducted in mice using male ICR mice (20-25g, fasted overnight). Three mice were administered 10 mg / kg orally via gavage. Blood samples were collected before administration and at 15, 30 minutes, and 1, 2, 4, 8, and 24 hours after administration. 6800g of blood samples were centrifuged at 2-8℃ for 6 minutes, and plasma was collected and stored at -80℃. Plasma samples from each time point were mixed with 3-5 times the volume of acetonitrile solution containing an internal standard, vortexed for 1 minute, centrifuged at 13000 rpm at 4℃ for 10 minutes, and the supernatant was mixed with 3 times the volume of water. An appropriate amount of the mixture was analyzed by LC-MS / MS. Key pharmacokinetic parameters were analyzed using a non-compartmental model with WinNonlin 7.0 software.

[0444] The experimental results show that, in mouse models, the pharmacokinetic properties of the compound of formula I of this invention are improved compared with those of control compounds 1 and 2.

[0445] Table 32. Pharmacokinetics of the tested compounds in mice.

[0446]

[0447] III. Taste Test in Rats

[0448] SD rats were trained to fast from water overnight for 3 days before being administered the drug. Half an hour after drug administration, each animal was given one bottle of water and one bottle of 0.3 mM quinine solution, respectively. The water bottles were removed 15 minutes after water administration, and the rats' water intake and 0.3 mM quinine intake were measured. The difference between water intake and quinine intake was then analyzed. Figure 41 The effects of the compound on taste in SD rats were evaluated.

[0449] Table 33 shows the water / water quinine uptake multiples in animals treated with the corresponding compounds.

[0450] compound Water / Quinine Water solvent 34.7 Control compound 1 30 mg / kg ip 11.4 Compound of Formula I 30 mg / kg ip 29.4

[0451] IV. Guinea pig histamine / citric acid cough efficacy test

[0452] Before being introduced into the group, the animals were acclimatized for 3-7 days. Once they reached the target weight (300-400g), they were numbered and randomly grouped.

[0453] The compound or excipient was administered to guinea pigs via nasal drops 0.25–24 hours prior to the start of cough assessment. The dosage range of the test substance was 0.17 mg / kg–1.5 mg / kg. During cough assessment, after the animals were acclimatized in a whole-body volumetric scanning chamber, they underwent histamine nebulization, followed by citric acid nebulization. The number of coughs and the cough latency were recorded for 22 minutes from the start of histamine nebulization until the end of the observation period.

[0454] The experimental data were analyzed and compared using one-way ANOVA. A p-value < 0.05 was considered statistically significant. Pairwise comparisons were performed using the t-test method.

[0455] Table 34 Number of coughs in guinea pigs after administration of the corresponding compounds, stimulated by histamine / citric acid.

[0456] compound Average number of coughs within 15 minutes Cough suppression rate vs. solvent (%) solvent 26.6 -- Reference compound 1 11.4 57.1 Formula I compound 8.9 66.5

[0457] Data showed that, compared with control compound 1, the compound of the present invention significantly reduced the number of coughs and prolonged the cough latency in a citric acid / histamine-induced guinea pig cough model, demonstrating a good antitussive effect. Figure 42 ).

[0458] V. Efficacy test of ATP / citric acid cough medicine in guinea pigs

[0459] Before enrollment, animals were acclimatized for 3-7 days until they reached the target weight (300-400g), at which point they were numbered and randomly grouped. The compound or excipient was administered to guinea pigs via nasal drops 0.25–24 hours before the cough assessment. The dosage range of the test substance was 0.17 mg / kg–1.5 mg / kg. During the cough assessment, after acclimatization in a whole-body volume scanning chamber, animals underwent ATP nebulization, followed by citric acid nebulization after a few minutes. The number of coughs and the cough latency were recorded within 15 minutes of the start of citric acid nebulization.

[0460] One-way ANOVA was used to analyze and compare the data of each group. A p-value < 0.05 was considered statistically significant. The t-test was used to compare differences between pairs of data.

[0461] Table 35 Number of coughs in guinea pigs after administration of the corresponding compounds, stimulated by ATP / citric acid.

[0462] compound Average number of coughs within 15 minutes Cough suppression rate vs. solvent (%) solvent 19.8 -- Reference compound 1 8.1 59.1 Formula I compound 6.3 68.2

[0463] Data showed that, compared with control compound 1, the compound of the present invention significantly reduced the number of coughs and prolonged the cough latency in a citric acid / ATP-stimulated guinea pig cough model, demonstrating a good antitussive effect. Figure 43 ).

[0464] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The crystal form of the compound of formula I, wherein, The structure of the compound of formula I is shown below: Formula I; The crystal form is the free alkali crystal form A of the compound of formula I. The X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.44°, 14.87°, 15.77°, 17.81°, and 18.61°.

2. The crystal form according to claim 1, wherein, The X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.44°, 11.14°, 11.36°, 14.87°, 15.77°, 16.97°, 17.81°, and 18.61°.

3. The crystal form according to claim 1, wherein, The X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 3.75°, 7.44°, 11.14°, 11.36°, 11.98°, 12.25°, 14.87°, 15.77°, 16.97°, 17.81°, 18.61°, and 22.36°.

4. The crystal form according to claim 1, wherein, The X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 3.75°, 5.99°, 7.44°, 9.01°, 9.93°, 11.14°, 11.36°, 11.98°, 12.25°, 13.88°, 14.20°, 14.87°, 15.77°, 16.97°, 17.81°, 18.61°, 22.36°, and 24.07°.

5. The crystal form according to claim 1, wherein, The X-ray powder diffraction pattern of the free alkali crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 3.75°, 5.99°, 7.44°, 9.01°, 9.93°, 11.14°, 11.36°, 11.98°, 12.25°, 13.88°, 14.20°, 14.87°, 15.77°, 16.97°, 17.81°, 18.61°, 19.39°, 20.26°, 21.14°, 22.36°, 23.34°, 24.07°, 26.33°, 26.78°, 27.18°, 28.17°, 30.20°, 33.88°, 34.35°, 37.23°, and 37.70°.

6. The crystal form according to claim 1, wherein, The free alkali crystal form A has a basic XRPD spectrum as shown in Figure 1-1; And / or, the free alkali crystal form A has one, two, or three of the following characteristics: (1) The TGA curve of free alkali crystal form A showed a weight loss of 1.28±1% at 150.0±3℃; (2) The DSC curve of free alkali crystal form A has an endothermic peak starting point at 175.6±3℃; (3) The DSC curve of free alkali crystal form A has an endothermic peak at 176.4±3℃.

7. The crystal form of the compound of formula I, wherein, The structure of the compound of formula I is shown below: Formula I; The crystal form is the free base crystal form B of the compound of formula I, wherein the X-ray powder diffraction pattern of the free base crystal form B, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 7.21°, 12.48°, 13.17°, 14.41°, 19.09°, 19.56°, 22.09°, and 26.49°.

8. The crystal form according to claim 7, wherein, The X-ray powder diffraction pattern of the free alkali crystal form B, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.21°, 12.48°, 13.17°, 14.41°, 16.72°, 19.09°, 19.56°, 20.90°, 22.09°, and 26.49°.

9. The crystal form according to claim 7, wherein, The X-ray powder diffraction pattern of the free alkali crystal form B, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.21°, 8.35°, 12.48°, 13.17°, 14.41°, 15.05°, 16.72°, 17.80°, 18.39°, 19.09°, 19.56°, 20.90°, 21.67°, 22.09°, 22.97°, 25.16°, 26.49°, and 27.49°.

10. The crystal form according to claim 7, wherein, The X-ray powder diffraction pattern of the free alkali crystal form B, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.21°, 8.35°, 12.48°, 13.17°, 14.41°, 15.05°, 16.72°, 17.80°, 18.39°, 19.09°, 19.56°, 20.90°, 21.67°, 22.09°, 22.97°, 25.16°, 25.45°, 26.49°, 27.49°, 28.66°, 29.10°, 29.35°, 31.71°, 32.00°, 32.85°, 33.70°, 34.23°, 36.78°, 38.26°, and 38.70°.

11. The crystal form according to claim 7, wherein, The free alkali crystal form B has an XRPD spectrum as shown in Figure 2-1; And / or, the free alkali crystal form B has one, two, or three of the following characteristics: (1) The TGA curve of free alkali crystal form B showed a weight loss of 2.36±1% at 150.0±3℃; (2) The DSC curve of free alkali crystal form B has an endothermic peak starting point at 177.0±3℃; (3) The DSC curve of free alkali crystal B has an endothermic peak at 179.4±3℃.

12. A pharmaceutically acceptable salt of compound I, wherein, The structure of the compound of formula I is shown below: Formula I; Pharmaceutically acceptable salts of the compound of formula I include salts formed by the compound of formula I with inorganic or organic acids; The inorganic acids include: hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, and hydrobromic acid; The organic acids include: maleic acid, L-aspartic acid, fumaric acid, L-tartaric acid, citric acid, D-glucuronic acid, L-malic acid, hippuric acid, D-gluconic acid, DL-lactic acid, succinic acid, L-ascorbic acid, adipic acid, acetic acid, p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, oxalic acid, 2-hydroxyethanesulfonic acid, malonic acid, gentian acid, and benzoic acid.

13. A pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, Pharmaceutically acceptable salts of the compound of formula I are hydrochloride, maleate, p-toluenesulfonate, benzenesulfonate, and malonate of the compound of formula I.

14. A pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, The molar ratio of the compound of Formula I to the acid is 5:1 to 1:

5.

15. A pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, The molar ratio of the compound of Formula I to the acid is 3:1, 2:1, 1:1, 1:1.5, 1:2, 1:2.5, or 1:

3.

16. The crystal form of a pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, The pharmaceutically acceptable salt of the compound of formula I is crystal form A of the hydrochloride salt of the compound of formula I. The X-ray powder diffraction pattern of the hydrochloride salt crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.77°, 9.01°, 10.10°, 15.54°, 17.51°, 19.24°, and 24.49°. The molar ratio of compound I in hydrochloric acid to hydrochloric acid in the hydrochloric acid crystal form A is 2:

1.

17. The crystal form according to claim 16, wherein, The X-ray powder diffraction pattern of the hydrochloride crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.77°, 9.01°, 10.10°, 15.54°, 17.51°, 18.01°, 19.24°, 20.05°, 21.28°, 23.38°, 23.79°, 24.49°, 26.07°, and 28.33°.

18. The crystal form according to claim 16, wherein, The X-ray powder diffraction pattern of the hydrochloride crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 7.77°, 9.01°, 10.10°, 15.54°, 17.51°, 18.01°, 19.24°, 20.05°, 21.28°, 21.59°, 22.67°, 23.38°, 23.79°, 24.49°, 26.07°, 27.17°, and 28.33°.

19. The crystal form according to any one of claims 16-18, wherein, The hydrochloride crystal form A has a basic XRPD spectrum as shown in Figure 3-1; And / or, the hydrochloride crystal form A has a VT-XRPD pattern as shown in Figure 9-4; And / or, the hydrochloride crystal form A has one, two, three, four, five, or six of the following characteristics: (1) The TGA curve of hydrochloride crystal form A shows a weight loss of 2.19±1% ​​at 100.0±3℃; (2) The TGA curve of hydrochloride crystal form A shows a weight loss of 3.90±1% in the temperature range of 100.0±3℃ to 160.0±3℃; (3) The DSC curve of hydrochloride crystal form A has an endothermic peak starting point at 143.6±3℃; (4) The DSC curve of hydrochloride crystal form A has an endothermic peak at 157.4±3℃; (5) The DSC curve of hydrochloride crystal form A has an endothermic peak at 176.0±3℃; (6) The DSC curve of hydrochloride crystal form A has an endothermic peak at 179.0±3℃.

20. The crystal form of a pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, The pharmaceutically acceptable salt of the compound of Formula I is maleate crystal form A, and the X-ray powder diffraction pattern of maleate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 6.73°, 10.84°, 14.68°, 16.26°, 18.23°, and 18.44°. The molar ratio of maleate crystal form A, compound of formula I, to maleic acid is 2:

1.

21. The crystal form according to claim 20, wherein, The X-ray powder diffraction pattern of the maleate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.43°, 6.73°, 10.84°, 14.68°, 16.26°, 16.82°, 18.23°, and 18.44°.

22. The crystal form according to claim 20, wherein the X-ray powder diffraction pattern of the maleate crystal form A, expressed as a diffraction angle of 2θ ± 0.2°, has diffraction peaks at 5.43°, 6.73°, 9.95°, 10.84°, 11.75°, 13.50°, 14.68°, 16.26°, 16.82°, 18.23°, 18.44°, 20.17°, 22.79°, 23.22°, 24.00°, 26.07°, 27.72°, and 28.86°.

23. The crystal form according to any one of claims 20-22, wherein, The maleate crystal form A has a basic XRPD spectrum as shown in Figure 4-1; And / or, the maleate crystal form A has a VT-XRPD pattern as shown in Figure 10-4; And / or, the maleate crystal form A has one, two, three or more of the following characteristics: (1) The TGA curve of maleate crystal form A shows a weight loss of 1.65±1% at 110.0±3℃; (2) The TGA curve of maleate crystal form A shows a weight loss of 11.88±1% in the temperature range of 110.0±3℃ to 220.0±3℃; (3) The DSC curve of maleate crystal form A has an endothermic peak at 107.8±3℃; (4) The DSC curve of maleate crystal form A has an endothermic peak starting point at 143.4±3℃; (5) The DSC curve of maleate crystal form A has an endothermic peak at 144.1±3℃; (6) The DSC curve of maleate crystal form A has an endothermic peak at 160.2±3℃.

24. The crystal form of a pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, The pharmaceutically acceptable salt of the compound of Formula I is p-toluenesulfonate crystal form A, and the X-ray powder diffraction pattern of p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 4.99°, 7.26°, 8.70°, 8.87°, 15.40°, 17.73°, 21.01°, and 24.13°. The molar ratio of compound I in p-toluenesulfonate crystal form A to p-benzenesulfonic acid is 1:

1.

25. The crystal form according to claim 24, wherein, The X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 4.99°, 7.26°, 8.70°, 8.87°, 15.20°, 15.40°, 16.68°, 17.73°, 19.71°, 21.01°, and 24.13°.

26. The crystal form according to claim 24, wherein, The X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 4.99°, 7.26°, 8.70°, 8.87°, 14.45°, 14.88°, 15.20°, 15.40°, 16.41°, 16.68°, 17.45°, 17.73°, 19.16°, 19.71°, 20.66°, 21.01°, 21.76°, 22.41°, 24.13°, 25.76°, 26.18°, and 27.25°.

27. The crystal form according to claim 24, wherein, The X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 4.99°, 7.26°, 8.70°, 8.87°, 14.45°, 14.88°, 15.20°, 15.40°, 16.41°, 16.68°, 17.45°, 17.73°, 19.16°, 19.71°, 20.66°, 21.01°, 21.76°, 22.41°, 24.13°, 25.76°, 26.18°, 27.25°, 27.95°, 29.23°, 30.69°, 31.00°, 31.78°, and 38.41°.

28. The crystal form according to any one of claims 24-27, wherein, The p-toluenesulfonate crystal form A has a basic XRPD spectrum as shown in Figure 5-1; And / or, the p-toluenesulfonate crystal form A has one, two, or three of the following characteristics: (1) The TGA curve of p-toluenesulfonate crystal form A showed a weight loss of 0.73±1% at 150.0±3℃; (2) The DSC curve of p-toluenesulfonate crystal form A has an endothermic peak starting point at 157.1±3℃; (3) The DSC curve of p-toluenesulfonate crystal form A has an endothermic peak at 159.2±3℃.

29. The crystal form of a pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, The pharmaceutically acceptable salt of the compound of Formula I is a benzenesulfonate crystal form A, and the X-ray powder diffraction pattern of the benzenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.36°, 7.28°, 8.34°, 9.64°, 16.20°, 18.55°, and 21.49°. The molar ratio of compound of formula I in benzenesulfonate crystal form A to benzenesulfonic acid is 1:

1.

30. The crystal form according to claim 29, wherein, The X-ray powder diffraction pattern of the benzenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.36°, 7.28°, 8.34°, 9.64°, 16.20°, 18.55°, 19.28°, 21.49°, 21.81°, 23.21°, 25.05°, and 25.74°.

31. The crystal form according to claim 29, wherein, The X-ray powder diffraction pattern of the benzenesulfonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.36°, 7.28°, 8.34°, 9.64°, 10.66°, 14.55°, 15.00°, 16.20°, 16.93°, 17.85°, 18.55°, 19.28°, 19.74°, 20.80°, 21.49°, 21.81°, 23.21°, 23.68°, 23.98°, 25.05°, 25.74°, 26.65°, and 27.82°.

32. The crystal form according to any one of claims 29-31, wherein, The benzenesulfonate crystal form A has a basic XRPD spectrum as shown in Figure 6-1; And / or, the benzenesulfonate crystal form A has one, two, or three of the following characteristics: (1) The TGA curve of benzenesulfonate crystal form A showed a weight loss of 1.35±1% at 120.0±3℃; (2) The DSC curve of benzenesulfonate crystal form A has an endothermic peak starting point at 159.6±3℃; (3) The DSC curve of benzenesulfonate crystal form A has an endothermic peak at 160.9±3℃.

33. The crystal form of a pharmaceutically acceptable salt of the compound of formula I according to claim 12, wherein, The pharmaceutically acceptable salt of the compound of Formula I is a malonate crystal form A, and the X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 6.75°, 9.96°, 10.67°, 14.48°, 16.04°, 16.88°, 18.04°, and 18.29°.

34. The crystal form according to claim 33, wherein, The X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.34°, 6.75°, 9.96°, 10.67°, 11.83°, 13.49°, 14.48°, 16.04°, 16.88°, 17.04°, 18.04°, 18.29°, and 27.38°.

35. The crystal form according to claim 33, wherein, The X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.34°, 6.75°, 9.96°, 10.67°, 11.83°, 13.49°, 14.48°, 16.04°, 16.88°, 17.04°, 18.04°, 18.29°, 20.27°, 22.57°, 22.95°, 27.38°, and 28.83°.

36. The crystal form according to claim 33, wherein, The X-ray powder diffraction pattern of the malonate crystal form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 5.34°, 6.75°, 9.96°, 10.67°, 11.83°, 13.49°, 14.48°, 16.04°, 16.88°, 17.04°, 18.04°, 18.29°, 18.63°, 20.27°, 21.57°, 22.57°, 22.95°, 24.11°, 24.83°, 26.02°, 27.38°, and 28.83°.

37. The crystal form according to any one of claims 33-36, wherein, The malonate crystal form A has a basic XRPD spectrum as shown in Figure 7-1; And / or, the malonate crystal form A has a basic VT-XRPD pattern as shown in Figure 7-4; And / or, the malonate crystal form A has one, two, three, or four of the following characteristics: (1) The TGA curve of malonate crystal form A shows a weight loss of 2.99±1% at 120.0±3℃; (2) The TGA curve of malonate crystal form A showed a weight loss of 11.02±1% in the temperature range of 120.0±3℃ to 200.0±3℃; (3) The DSC curve of malonate crystal form A has an endothermic peak starting point at 155.6±3℃; (4) The DSC curve of malonate crystal form A has an endothermic peak at 156.4±3℃; (5) The DSC curve of malonate crystal form A has an endothermic peak at 172.4±3℃.

38. A method for preparing the free alkali crystal form A of the compound of formula I according to any one of claims 1-6, wherein, Choose any one of the following methods: Method 1: Add compound I to organic solvent I, dissolve, filter, and volatilize at room temperature. The organic solvent I is selected from one or more of acetone, tetrahydrofuran, dichloromethane, acetonitrile, and ethyl acetate; Method 2: Dissolve the compound of formula I completely in organic solvent II, and add the antisolvent dropwise to the clear solution under stirring until a solid precipitates; if no solid precipitates, use suspension stirring; if no solid still precipitates, cool down and suspend stirring, then allow the clear solution to evaporate at room temperature. The organic solvent II is selected from one or more of methanol, acetone, ethyl acetate, tetrahydrofuran, chloroform, N,N-dimethylacetamide, and N-methylpyrrolidone; The antisolvent is selected from one or more of water, m-xylene, n-hexane, cumene, toluene, cyclohexane, n-heptane, n-pentane, and p-isopropyltoluene; Method 3: Place the first sample vial containing compound I openly into a second sample vial containing solvent, seal the second sample vial, and let it stand at room temperature; the solvent should not cover the mouth of the first sample vial. The solvent is selected from one or more of ethanol, dichloromethane, acetonitrile, acetone, toluene, N,N-dimethylacetamide, and n-hexane; Method 4: Place the first sample vial containing the solution of compound I openly into the second sample vial containing the antisolvent, seal the second sample vial, and let it stand at room temperature; the antisolvent does not submerge the mouth of the first sample vial. The solvent in the compound solution of Formula I is selected from one or more of isopropanol, methyl isobutyl ketone, 1,4-dioxane, and dimethyl sulfoxide. The antisolvent is selected from one or more of n-pentane, methyl butyl ether, water, and m-xylene; Method 5: Add the polymer to the solution of compound I and allow it to evaporate at room temperature; The solvent in the solution of the compound of Formula I is selected from one or more of methanol, 2-butanone, methyl acetate, isopropyl acetate, ethanol, dichloromethane, and 2-methyltetrahydrofuran; The polymer is selected from one or more of polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose, methylcellulose, polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate, and hydroxyethyl cellulose. Method Six: Place the first sample vial containing the compound of Formula I open into a second sample vial containing a saturated salt solution or water, seal the second sample vial, and let it stand at room temperature; the solvent should not cover the mouth of the first sample vial. The saturated salt solution is a saturated inorganic salt solution; The inorganic salt saturated solution is a saturated potassium acetate solution, a saturated potassium carbonate solution, a saturated sodium bromide solution, or a saturated potassium bromide solution. The system's humidity is 15~100% RH; Method 7: The suspension of compound I is stirred and centrifuged under temperature cycling, and the solid is collected; The solvent in the suspension is selected from one or more of the following: n-heptane, methyl butyl ether, anisole, dicyclohexylamine, acetone, ethanol, ethyl acetate, methylcyclohexane, chloroform, 2-butanone, m-xylene, and water. The temperature cycling conditions include: 50 ºC~5 ºC, 0.1~0.5 ºC / min, and at least 2 cycles; Method 8: Dissolve the compound of formula I completely in the positive solvent, and add the antisolvent dropwise to the clear solution under stirring until a solid precipitates; if no solid precipitates, use suspension stirring; if no solid still precipitates, cool down and suspend stirring, then allow the clear solution to evaporate at room temperature; The positive solvent is selected from one or more of ethanol, ethyl acetate, 2-methyltetrahydrofuran, 2-butanone, acetonitrile, dichloromethane, and 1,4-dioxane; The antisolvent is selected from one or more of n-heptane, tetrahydrofuran, and water; Method 9: Place the turbid liquid of compound I at room temperature, stir magnetically, centrifuge, and collect the solid; The solvent of the turbid liquid is selected from one or more of the following: isobutanol, methyl tert-butyl ether, cyclohexane, toluene, isopropyl acetate, water, methylcyclohexane, tetrahydrofuran, n-pentane, acetone, isopropanol, cyclopentyl methyl ether, methanol, p-isopropyltoluene, dichloromethane, n-heptane, acetonitrile, 1,4-dioxane, and N-methylpyrrolidone. Method 10: Weigh the compound of Formula I into an HPLC bottle, add solvent to the HPLC bottle, heat and stir to equilibrate, then filter to obtain the supernatant; place the supernatant in a biological incubator, cool it from 50 ºC to 5 ºC at a rate of 0.05 ºC / min, and then maintain the temperature at 5 ºC; transfer the clear solution to -20 ºC and maintain the temperature thereafter, collect the precipitated solid, and transfer the sample without precipitated solid to room temperature for volatilization; The solvent is selected from one or more of isopropanol, anisole, isopropyl acetate, tetrahydrofuran, and water; The temperature to be heated is 45-55 ºC; Method 11: Stir the suspension of compound I, centrifuge, and collect the solid; The solvent in the suspension is selected from one or more of the following: n-butanol, toluene, diisopropyl ether, methylcyclohexane, cumene, anisole, water, petroleum ether, dicyclohexylamine, 2-methyltetrahydrofuran, n-hexane, 2-butanone, isopropyl acetate, chloroform, m-xylene, tetrahydrofuran, methyl isobutyl ketone, cyclopentyl methyl ether, and benzyl alcohol. The suspension was stirred at 45-55ºC.

39. A method for preparing the free alkali crystal form B of the compound of formula I according to any one of claims 7-11, wherein, Includes the following steps: The free acid crystal form A of the compound of formula I according to any one of claims 1-6 is dissolved in 1,4-dioxane and then subjected to gas-liquid diffusion in a hexane atmosphere to obtain the free base crystal form B of the compound of formula I.

40. A method for preparing a pharmaceutically acceptable salt of the compound of formula I according to any one of claims 12-15, comprising the steps of: mixing the compound of formula I or the free base crystal form A of the compound of formula I according to any one of claims 1-6 with a salt-forming reagent in a suitable solvent to obtain a mixture; wherein the salt-forming reagent is an inorganic acid or an organic acid.

41. A pharmaceutical composition, wherein, The pharmaceutical composition comprises one or more of the crystal forms of the compound of formula I according to any one of claims 1-11, pharmaceutically acceptable salts of the compound of formula I according to any one of claims 12-15, or pharmaceutically acceptable salts of the compound of formula I according to any one of claims 16-37.

42. The pharmaceutical composition according to claim 41, wherein, The pharmaceutically acceptable salts of the compound of formula I are hydrochloride, maleate, p-toluenesulfonate, benzenesulfonate, and malonate of the compound of formula I.

43. Use of the crystal form of the compound of formula I according to any one of claims 1-11, a pharmaceutically acceptable salt of the compound of formula I according to any one of claims 12-15, a crystal form of a pharmaceutically acceptable salt of the compound of formula I according to any one of claims 16-37, or the use of the pharmaceutical composition according to any one of claims 41-42 in the preparation of a medicament for treating and / or preventing P2X3-related diseases.

44. The use according to claim 43, wherein, The P2X3-related diseases are selected from: pain, reproductive and urinary system diseases, or respiratory system diseases.

45. The use according to claim 44, wherein, The pain is selected from: inflammatory pain, surgical pain, visceral pain, toothache, premenstrual pain, central pain, pain caused by burns, migraine or cluster headache; The reproductive and urinary system diseases mentioned are selected from: urinary incontinence, overactive bladder, dysuria, cystitis, endometriosis, and endometriosis-related pain; The respiratory diseases mentioned are selected from: cough, idiopathic pulmonary fibrosis, and chronic obstructive pulmonary disease.

46. ​​The use according to claim 45, wherein the cough is selected from subacute or chronic cough.

47. The use according to claim 45, wherein the cough is selected from the treatment of resistant cough.

48. The use according to claim 45, wherein the cough is selected from idiopathic chronic cough.

49. The use according to claim 45, wherein the cough is selected from post-viral infection cough or iatrogenic cough.

50. The use according to claim 45, wherein the cough is selected from coughs related to respiratory diseases.

51. A method for quality control of the crystal form of a compound of formula I according to any one of claims 1-11 or the crystal form of a pharmaceutically acceptable salt of a compound of formula I according to any one of claims 16-37, comprising detecting the content of the crystal form using high performance liquid chromatography; wherein the mobile phase used in the high performance liquid chromatography includes mobile phase A and mobile phase B; The mobile phase A is an aqueous solution of formic acid and acetonitrile; the mobile phase B is acetonitrile.

52. The quality inspection method according to claim 51, wherein, The mobile phase A is an aqueous solution of 0.05-0.15% formic acid and 2-7% acetonitrile; And / or, the high performance liquid chromatography method employs gradient elution; And / or, the flow rate of the mobile phase is 1 ± 0.2 mL / min; the gradient elution time is 5-60 min; And / or, in the gradient elution, the volume ratio of mobile phase A to mobile phase B is 1:9-9:

1.

53. The quality inspection method according to claim 51 or 52, wherein, The quality testing methods include: purity testing methods, solubility testing methods, and stability testing methods.

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