7,8-Dihydroxyflavone crystalline polymorphs and methods for their preparation

Abstract

This invention relates to crystalline polymorphs of 7,8-dihydroxyflavone and methods for their preparation. [Means] The 7,8-dihydroxyflavone crystalline polymorph has significantly improved physicochemical properties, thereby possessing higher pharmaceutical value.

Description

Technical Field

[0001] The present disclosure relates to crystal polymorphs of 7,8-dihydroxyflavone and a method for preparing the same.

Background Art

[0002] As a specific agonist of tyrosine receptor kinase B (TrkB), 7,8-dihydroxyflavone can activate TrkB. According to many in vitro studies, 7,8-dihydroxyflavone has important biological effects, and mainly has good therapeutic effects in neurodegenerative diseases including diseases such as Parkinson's disease, Alzheimer's disease, depression, psychotic disorders, post-traumatic stress disorder, autism spectrum disorder, stroke, Rett syndrome, etc.

[0003] The crystal polymorph phenomenon means that a substance has two or more different crystal structures, which is a phenomenon commonly seen in solid substances. In the pharmaceutical field, research on the crystal polymorph phenomenon of drugs has already become an important forefront issue. When different crystal structures exist in the same solid substance, they often exhibit different physicochemical properties. For drugs, different crystal forms mainly show differences in aspects such as stability, solubility, and dissolution rate, which affect the drug efficacy of the drug and ultimately change the toxic and side effects of the drug.

[0004] Therefore, the development of a crystal form of 7,8-dihydroxyflavone with excellent performance has very important significance.

Summary of the Invention

Problems to be Solved by the Invention

[0005] The present disclosure discloses crystal polymorphs of 7,8-dihydroxyflavone and a method for preparing the same. The crystal polymorphs of 7,8-dihydroxyflavone according to the present disclosure have more selectable pharmaceutical economic values.

Means for Solving the Problems

[0006] In one aspect, the present disclosure The X-ray powder diffraction pattern has characteristic peaks at approximately 8.3±0.2°, 25.0±0.2°, and 25.9±0.2° at a 2θ angle. The crystalline form P of 7,8-dihydroxyflavone is provided.

[0007] In one embodiment, the X-ray powder diffraction pattern further includes characteristic peaks at approximately 27.4 ± 0.2° and / or 31.4 ± 0.2°.

[0008] In one further embodiment, the X-ray powder diffraction pattern further includes characteristic peaks at approximately 12.5±0.2°, 15.8±0.2°, 18.4±0.2°, 18.7±0.2°, 19.4±0.2° and / or 33.7±0.2°.

[0009] In one further embodiment, the crystal form P has an X-ray powder diffraction pattern essentially as shown in Figure 1.

[0010] In one further embodiment, the thermogravimetric spectrum of the crystalline form P shows no apparent weight loss.

[0011] In one further embodiment, the differential scanning calorimetry spectrum of the crystalline form P does not have an endothermic signal corresponding to weight loss.

[0012] In one further embodiment, the dynamic moisture adsorption graph of the crystalline form P shows a weight increase of less than 0.1% in the range of relative humidity from 0% to 95%, where "%" is the percentage of the increased mass of the crystalline form P of 7,8-dihydroxyflavone relative to the initial mass.

[0013] In one further embodiment, the crystalline form P of the 7,8-dihydroxyflavone essentially has a thermogravimetric spectrum and / or differential scanning calorimetry spectrum as shown in Figure 4.

[0014] In one further embodiment, the crystalline form P of the 7,8-dihydroxyflavone basically has a dynamic water adsorption graph as shown in Figure 5.

[0015] In one further embodiment, the crystalline form P of the 7,8-dihydroxyflavone is an anhydrous form.

[0016] In another aspect, this disclosure is: The X-ray powder diffraction pattern has characteristic peaks at approximately 11.1±0.2°, 15.6±0.2°, 27.0±0.2°, and 28.3±0.2° at 2θ angles. This provides the crystalline form C of 7,8-dihydroxyflavone.

[0017] In one embodiment, the X-ray powder diffraction pattern further includes characteristic peaks at approximately 5.6±0.2°, 9.5±0.2°, 15.5±0.2°, 21.0±0.2°, 24.3±0.2°, and / or 25.5±0.2°.

[0018] In one further embodiment, the X-ray powder diffraction pattern further includes characteristic peaks at 12.7±0.2°, 13.7±0.2°, 18.3±0.2°, 20.4±0.2°, 21.9±0.2°, 23.2±0.2°, 25.2±0.2° and / or 29.7±0.2°.

[0019] In one further embodiment, the crystal form C has an X-ray powder diffraction pattern essentially as shown in Figure 2.

[0020] In one further embodiment, in the thermogravimetric spectrum of the crystalline form C, the mass of the decrease between 110°C and 150°C accounts for approximately 3.7% of the mass before the decrease, where "%" is a mass percentage.

[0021] In one further embodiment, the differential scanning calorimetry spectrum of the crystalline form C has an endothermic signal corresponding to weight loss at approximately 100°C to 160°C.

[0022] In a further embodiment, in the dynamic moisture sorption graph of the crystalline form C, the weight increase is less than 0.1% within the range of relative humidity from 0% to 95%, and the “%” is the percentage of the increased mass of the crystalline form C of the 7,8-dihydroxyflavone occupying the initial mass.

[0023] In a further embodiment, the crystalline form C of the 7,8-dihydroxyflavone basically has a thermogravimetric analysis spectrum and / or a differential scanning calorimetry spectrum as shown in FIG. 6.

[0024] In a further embodiment, the crystalline form C of the 7,8-dihydroxyflavone basically has a dynamic moisture sorption graph as shown in FIG. 7.

[0025] In a further embodiment, the crystalline form C of the 7,8-dihydroxyflavone is a hydrate, for example, a monohydrate.

[0026] In another aspect, the present disclosure has an X-ray powder diffraction pattern including characteristic peaks shown at about 9.3 ± 0.2°, 23.8 ± 0.2°, 24.6 ± 0.2° and 26.6 ± 0.2° in 2θ angle, and provides a crystalline form N of 7,8-dihydroxyflavone.

[0027] In one embodiment, the X-ray powder diffraction pattern further includes characteristic peaks at about 12.3 ± 0.2° and / or 21.1 ± 0.2°.

[0028] In a further embodiment, the X-ray powder diffraction pattern further includes characteristic peaks at about 5.7 ± 0.2°, 11.9 ± 0.2°, 13.1 ± 0.2°, 14.2 ± 0.2°, 15.9 ± 0.2°, 16.8 ± 0.2°, 18.9 ± 0.2°, 20.7 ± 0.2° and / or 30.3 ± 0.2°.

[0029] In a further embodiment, the crystalline form N of the 7,8-dihydroxyflavone basically has an X-ray powder diffraction pattern as shown in FIG. 3.

[0030] In one further embodiment, in the thermogravimetric spectrum of the crystalline form N, the mass of the decrease between 40°C and 80°C accounts for 6.3% of the mass before the decrease, where "%" is a mass percentage.

[0031] In one further embodiment, the differential scanning calorimetry spectrum of the crystalline form N has an endothermic peak corresponding to weight loss at about 100°C and heat dissipation signals at about 136°C and 169°C, respectively.

[0032] In one further embodiment, the dynamic moisture adsorption graph of the crystalline form N shows a weight increase of less than 0.1% in the range of relative humidity from 0% to 95%, where "%" is the percentage of the increased mass of the crystalline form N of 7,8-dihydroxyflavone relative to the initial mass.

[0033] In one further embodiment, the crystalline form N of the 7,8-dihydroxyflavone has a thermogravimetric spectrum and / or differential scanning calorimetry spectrum, essentially as shown in Figure 8.

[0034] In one further embodiment, the crystalline form N of the 7,8-dihydroxyflavone basically has a dynamic water adsorption graph as shown in Figure 9.

[0035] In one further embodiment, the crystalline form N of the 7,8-dihydroxyflavone is a hydrate, for example, a dihydrate.

[0036] In another embodiment, the present disclosure provides a method for preparing a crystalline form of the 7,8-dihydroxyflavone of the present disclosure, which is one of Methods 1 to 5.

[0037] Method 1: This method comprises the steps of dissolving 7,8-dihydroxyflavone in a single solvent and volatilizing the solvent to obtain a solid.

[0038] Method 2: This method involves suspending 7,8-dihydroxyflavone in a single solvent or a two-component solvent, and then centrifuging it to obtain a solid.

[0039] Method 3: This method includes the steps of dissolving 7,8-dihydroxyflavone in a good solvent, adding the resulting solution dropwise to an antisolvent to precipitate a solid, and then separating the solid.

[0040] Method 4: This method includes the steps of dissolving 7,8-dihydroxyflavone in a good solvent, adding an antisolvent dropwise to the resulting solution to precipitate a solid, and then separating the solid.

[0041] Method 5 comprises the steps of suspending 7,8-dihydroxyflavone in an antisolvent, then adding a good solvent dropwise until the solid is dissolved, cooling to precipitate the solid, and then separating the solid.

[0042] In one embodiment, In Method 2, the single solvent is n-heptane, and the two-component solvent is a combination of one of 4-methyl-2-pentanone, n-propyl alcohol, N,N-dimethylformamide, ethylene glycol dimethyl ether, ethanol, and butyl formate, and one of isopropyl acetate, n-heptane, toluene, isopropyl alcohol, ethyl acetate, diethyl ether, and cyclohexane. and / or, in method 2, the volume ratio of the two-component solvent is 1:5 to 1:10. This invention provides a method for preparing crystalline form P of 7,8-dihydroxyflavone, characterized by the following features.

[0043] In one further embodiment, In Method 2, the temperature of the suspension is 20-60°C. This invention provides a method for preparing crystalline form P of 7,8-dihydroxyflavone, characterized by the following features.

[0044] In one embodiment, a 7,8-dihydroxyflavone having at least one feature of crystalline form P of the Disclosure, prepared by the method of the Disclosure, is provided.

[0045] In one embodiment, In Method 1, the single solvent is one of isopropyl acetate and N,N-dimethylformamide. and / or, in method 1, the volume mass ratio of the single solvent to 7,8-dihydroxyflavone is 5 to 500 mL / g, and / or, in method 1, the solid is precipitated by solvent evaporation, and / or, in method 2, the single solvent is water or diethyl ether, and the two-component solvent is selected from a combination of an organic solvent and water, a combination of butyl formate and n-heptane, a combination of ethylene glycol methyl ether and chloroform, and a combination of acetonitrile and toluene. and / or, in method 2, the organic solvent is one or more selected from tetrahydrofuran, isopropyl alcohol, acetonitrile, methanol, ethanol, dimethyl sulfoxide, N,N-dimethylformamide, and ethylene glycol dimethyl ether. and / or, in method 2, the two-component solvent has a volume ratio of organic solvent to water of 1:5 to 1:10, a volume ratio of butyl formate to n-heptane of 1:1, a volume ratio of ethylene glycol methyl ether to chloroform of 1:5, and / or a volume ratio of acetonitrile to toluene of 1:1. and / or, in method 3, the good solvent is one or more selected from N,N-dimethylformamide, tetrahydrofuran, and ethylene glycol dimethyl ether, and / or the antisolvent is one or more selected from water, toluene, methylene chloride, and diethyl ether. and / or, in method 3, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10, and / or, in method 4, the good solvent is one or more of N,N-dimethylformamide and isopropyl alcohol, and / or the antisolvent is one or more of chloroform, toluene and water. and / or, in method 4, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10. and / or, in method 5, the antisolvent is water, and the good solvent is one or more of N,N-dimethylformamide, tetrahydrofuran, ethanol, and ethylene glycol methyl ether. and / or, in method 5, the volume ratio of the antisolvent to the good solvent is 1:0.8 to 1:1. The present invention provides a method for preparing crystalline form C of 7,8-dihydroxyflavone, characterized by the above.

[0046] In one further embodiment, In Method 1, the single solvent is one of isopropyl acetate and N,N-dimethylformamide. and / or, in method 1, the temperature of solid precipitation is room temperature. and / or, in method 2, the suspension temperature is 20-60°C. And / or, in method 3, the dropping temperature is 15-50°C. And / or, in method 4, the dropping temperature is 15-50°C. And / or, in method 5, the cooling temperature is -15°C to 4°C. The present invention provides a method for preparing crystalline form C of 7,8-dihydroxyflavone, characterized by the above.

[0047] In one embodiment, a 7,8-dihydroxyflavone having at least one feature of crystalline form C of the Disclosure, prepared by the method of the Disclosure, is provided.

[0048] In one embodiment, In Method 1, the single solvent is methanol. and / or, in method 1, the volume mass ratio of the single solvent to 7,8-dihydroxyflavone is 5 to 500 mL / g, and / or, in method 1, the solid is precipitated by solvent evaporation, and / or, in method 2, the single solvent is methanol, and the two-component solvent is a combination of methanol and water. and / or, in method 2, the two-component solvent has a volume ratio of methanol to water of 1:5 to 1:10. and / or, in method 3, the good solvent is ethanol, and the antisolvent is one or more of methyl-t-butyl ether and toluene. and / or, in method 3, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10, and / or, in method 4, the good solvent is ethanol, and the antisolvent is one or more of cyclohexane, n-heptane, and water. and / or, in method 4, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10. and / or, in method 5, the antisolvent is one or more of toluene, isopropyl acetate, 4-methyl-2-pentanone, water, and acetonitrile, and the good solvent is one or more of ethanol and methanol. and / or, in method 5, the volume ratio of the antisolvent to the good solvent is 1:0.8 to 1:1. This invention provides a method for preparing crystalline form N of 7,8-dihydroxyflavone, characterized by the following features.

[0049] In one further embodiment, In Method 1, the single solvent is one of isopropyl acetate and N,N-dimethylformamide. and / or, in method 1, the temperature at which the solid is precipitated is room temperature. and / or, in method 2, the temperature of the suspension is 20-60°C. and / or, in method 3, the temperature of the drop is 15-50°C, and / or, in method 4, the temperature of the drop is 15-50°C, and / or, in method 5, the cooling temperature is -15 to 4°C. This invention provides a method for preparing crystalline form N of 7,8-dihydroxyflavone, characterized by the following features.

[0050] In one embodiment, a 7,8-dihydroxyflavone having at least one feature of crystalline form N of the present disclosure, prepared by the method of the present disclosure, is provided.

[0051] In another aspect, this disclosure is: (1) crystalline form P of 7,8-dihydroxyflavone, crystalline form C of 7,8-dihydroxyflavone and / or crystalline form N of 7,8-dihydroxyflavone according to the present disclosure, and (2) comprising at least one pharmaceutically acceptable excipient, To provide a pharmaceutical composition.

[0052] In another aspect, this disclosure is: A method for treating diseases associated with abnormalities in the TrkB signaling pathway, such as central nervous system injury, peripheral nerve injury, neurodegenerative lesions, psychiatric disorders, hereditary neurological disorders, ophthalmic diseases, metabolic diseases, pain, cardiovascular diseases, tumors, or other related diseases, (1) administering to a subject crystalline form P of 7,8-dihydroxyflavone, crystalline form C of 7,8-dihydroxyflavone, and / or crystalline form N of 7,8-dihydroxyflavone according to the present disclosure, and (2) at least one pharmaceutically acceptable excipient, Provide a method. [Brief explanation of the drawing]

[0053] [Figure 1] This is the X-ray powder diffraction pattern of the crystalline form P of 7,8-dihydroxyflavone obtained in Example 1. [Figure 2] This is the X-ray powder diffraction pattern of the crystalline form C of 7,8-dihydroxyflavone obtained in Example 2. [Figure 3]This is the X-ray powder diffraction pattern of crystalline form N of 7,8-dihydroxyflavone obtained in Example 3. [Figure 4] These are the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of the crystalline form P of 7,8-dihydroxyflavone obtained in Example 1. [Figure 5] This is a dynamic water adsorption graph of the crystalline form P of the 7,8-dihydroxyflavone obtained in Example 1. [Figure 6] These are the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of the crystalline form C of 7,8-dihydroxyflavone obtained in Example 2. [Figure 7] This is a dynamic water adsorption graph of crystalline form C of the 7,8-dihydroxyflavone obtained in Example 2. [Figure 8] These are the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of the crystalline form N of 7,8-dihydroxyflavone obtained in Example 3. [Figure 9] This is a dynamic water adsorption graph of the crystalline form N of 7,8-dihydroxyflavone obtained in Example 3. [Figure 10] These are the X-ray powder diffraction patterns of the crystalline form P of 7,8-dihydroxyflavone obtained in Example 1, before and after the dynamic moisture adsorption experiment. [Figure 11] This is a polarized light microscope image of the crystalline form P of the 7,8-dihydroxyflavone obtained in Example 1. [Figure 12] This is the nuclear magnetic hydrogen spectrum of the crystalline form P of the 7,8-dihydroxyflavone obtained in Example 1. [Figure 13] These are the X-ray powder diffraction patterns of the crystalline form C of 7,8-dihydroxyflavone obtained in Example 2, before and after the dynamic moisture adsorption experiment. [Figure 14] This is a polarized light microscope image of the crystalline form C of 7,8-dihydroxyflavone obtained in Example 2. [Figure 15] This is the nuclear magnetic hydrogen spectrum of the crystalline form C of the 7,8-dihydroxyflavone obtained in Example 2. [Figure 16]These are the X-ray powder diffraction patterns of the crystalline form N of 7,8-dihydroxyflavone obtained in Example 3, before and after the dynamic moisture adsorption experiment. [Figure 17] This is a polarized light microscope analysis image of crystalline form N of the 7,8-dihydroxyflavone obtained in Example 3. [Figure 18] This is the nuclear magnetic hydrogen spectrum of the crystalline form N of the 7,8-dihydroxyflavone obtained in Example 3. [Figure 19] This is the X-ray powder diffraction pattern of crystalline form A of 7,8-dihydroxyflavone obtained in Comparative Example 1. [Figure 20] This is the X-ray powder diffraction pattern of crystalline form B of 7,8-dihydroxyflavone obtained in Comparative Example 2. [Figure 21] This is the X-ray powder diffraction pattern of crystalline form E of 7,8-dihydroxyflavone obtained in Comparative Example 3. [Figure 22] This is the X-ray powder diffraction pattern of the crystalline form F of 7,8-dihydroxyflavone obtained in Comparative Example 4. [Figure 23] This is the X-ray powder diffraction pattern of crystalline form G of 7,8-dihydroxyflavone obtained in Comparative Example 5. [Figure 24] This is the X-ray powder diffraction pattern of the crystalline form H of 7,8-dihydroxyflavone obtained in Comparative Example 6. [Figure 25] This is the X-ray powder diffraction pattern of crystalline form I of 7,8-dihydroxyflavone obtained in Comparative Example 7. [Figure 26] This is the X-ray powder diffraction pattern of the crystalline form J of 7,8-dihydroxyflavone obtained in Comparative Example 8. [Figure 27] This is the X-ray powder diffraction pattern of the crystalline form K of 7,8-dihydroxyflavone obtained in Comparative Example 9. [Figure 28] This is the X-ray powder diffraction pattern of the crystalline form L of 7,8-dihydroxyflavone obtained in Comparative Example 10. [Figure 29] This is the X-ray powder diffraction pattern of the crystalline form M of 7,8-dihydroxyflavone obtained in Comparative Example 11. [Figure 30] This is the X-ray powder diffraction pattern of the crystalline form O of 7,8-dihydroxyflavone obtained in Comparative Example 12. [Figure 31] This is the X-ray powder diffraction pattern of the crystalline form Q of 7,8-dihydroxyflavone obtained in Comparative Example 13. [Figure 32] This is the X-ray powder diffraction pattern of the crystalline form R of 7,8-dihydroxyflavone obtained in Comparative Example 14. [Figure 33] This is the X-ray powder diffraction pattern of the crystalline form S of 7,8-dihydroxyflavone obtained in Comparative Example 15. [Figure 34] This is the X-ray powder diffraction pattern of the crystalline form T of 7,8-dihydroxyflavone obtained in Comparative Example 16. [Figure 35] This is the X-ray powder diffraction pattern of the crystalline form P of 7,8-dihydroxyflavone obtained in Effect Example 1. [Figure 36] This is the X-ray powder diffraction pattern of crystalline form C of 7,8-dihydroxyflavone obtained in Effect Example 1. [Figure 37] This is the X-ray powder diffraction pattern of crystalline form N of 7,8-dihydroxyflavone obtained in Effect Example 1. [Figure 38] This is the X-ray powder diffraction pattern of the crystalline form P of 7,8-dihydroxyflavone obtained in Effect Example 2. [Figure 39] This is the X-ray powder diffraction pattern of the crystalline form C of 7,8-dihydroxyflavone obtained in Effect Example 2. [Figure 40] This is the X-ray powder diffraction pattern of crystalline form N of 7,8-dihydroxyflavone obtained in Effect Example 2. [Modes for carrying out the invention]

[0054] In this disclosure, the structural formula of the 7,8-dihydroxyflavone is as follows: [ka]

[0055] Unless otherwise explicitly stated, the term 7,8-dihydroxyflavone as used in this disclosure is not required to have any particular physical state and may be amorphous or in any crystalline form.

[0056] In one embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form P, characterized by its X-ray diffraction pattern having X-ray diffraction peaks at 2θ values ​​of approximately 8.3±0.2°, 25.0±0.2°, and 25.9±0.2°. In one further embodiment, the X-ray diffraction pattern of crystalline form P also has X-ray diffraction peaks at 2θ values ​​of approximately 27.4±0.2° and / or 31.4±0.2°. In one further embodiment, crystalline form P is anhydrous. In one further embodiment, the X-ray diffraction pattern of crystalline form P is as shown in Figure 1. In one further embodiment, the locations of characteristic X-ray diffraction peaks of crystalline form P are shown in Table 1 along with the locations of characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone.

[0057] In one embodiment, the present disclosure relates to a crystalline form P of 7,8-dihydroxyflavone that is stable and does not undergo crystallization.

[0058] In one embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form C, characterized by its X-ray diffraction pattern having X-ray diffraction peaks at 2θ values ​​of approximately 5.6±0.2°, 9.5±0.2°, 11.1±0.2°, 15.5±0.2°, 15.6±0.2°, 21.0±0.2°, 24.3±0.2°, 25.5±0.2°, 27.0±0.2°, and 28.3±0.2°. In one further embodiment, crystalline form C is a hydrate, e.g., a monohydrate. In one further embodiment, the X-ray diffraction pattern of crystalline form C is shown in Figure 2. In one further embodiment, the positions of characteristic X-ray diffraction peaks of crystalline form C are shown in Table 1 along with the positions of characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone.

[0059] In one effective embodiment, the present disclosure relates to a crystalline form C of 7,8-dihydroxyflavone that does not undergo a stable crystal transition.

[0060] In one embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form N, characterized by its X-ray diffraction pattern having X-ray diffraction peaks at 2θ values ​​of approximately 9.3±0.2°, 23.8±0.2°, 24.6±0.2°, and 26.6±0.2°. In one further embodiment, the X-ray diffraction pattern of crystalline form N also has X-ray diffraction peaks at 2θ values ​​of approximately 12.3±0.2° and / or 21.1±0.2°. In one further embodiment, crystalline form N is a hydrate, e.g., a dihydrate. In one further embodiment, the X-ray diffraction pattern of crystalline form N is shown in Figure 3. In one further embodiment, the locations of characteristic X-ray diffraction peaks of crystalline form N are shown in Table 1 along with the locations of characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone.

[0061] In one embodiment, the present disclosure relates to a crystalline form N of 7,8-dihydroxyflavone that is stable and does not undergo crystallization transition.

[0062] This disclosure relates to a method for preparing crystalline forms of 7,8-dihydroxyflavone, including Methods 1 to 5.

[0063] Method 1: This method involves dissolving 7,8-dihydroxyflavone in a single solvent and obtaining a solid until the solvent has completely evaporated.

[0064] Method 2: This method involves suspending 7,8-dihydroxyflavone in a single solvent or a two-component solvent, and then centrifuging it to obtain a solid.

[0065] Method 3: This method includes the steps of dissolving 7,8-dihydroxyflavone in a good solvent, adding it dropwise to an antisolvent, precipitating the solid, and then centrifuging to obtain the solid.

[0066] Method 4: This method involves dissolving 7,8-dihydroxyflavone in a good solvent, adding an antisolvent dropwise thereto, precipitating a solid, and then centrifuging to obtain the solid.

[0067] Method 5 comprises the steps of suspending 7,8-dihydroxyflavone in an antisolvent, adding a good solvent dropwise until the solid is dissolved, cooling to precipitate the solid, and then centrifuging to obtain the solid.

[0068] In one embodiment, the present disclosure relates to a method for preparing 7,8-dihydroxyflavone having at least one crystalline form P, in particular a pure crystalline form P. The method may be, for example, as follows:

[0069] In Method 2, the single solvent is n-heptane, and the two-component solvent is a combination of one of 4-methyl-2-pentanone, n-propyl alcohol, N,N-dimethylformamide, ethylene glycol dimethyl ether, ethanol, and butyl formate, and one of isopropyl acetate, n-heptane, toluene, isopropyl alcohol, ethyl acetate, diethyl ether, and cyclohexane. More preferably, the single solvent is n-heptane, and the two-component solvent is a combination of ethanol and ethyl acetate. and / or, in method 2, the volume ratio of the two-component solvent is 1:5 to 1:10, more preferably 1:10. In Method 2, the temperature of the suspension is 20 to 60°C, and more preferably, the temperature of the suspension is 50°C.

[0070] In one embodiment, the present disclosure relates to a method for preparing 7,8-dihydroxyflavone having at least one crystalline form C, in particular pure crystalline form C. The method may be, for example, as follows:

[0071] In Method 1, the single solvent is one of isopropyl acetate and N,N-dimethylformamide, and more preferably the single solvent is N,N-dimethylformamide. and / or, in method 1, the volume mass ratio of the single solvent to 7,8-dihydroxyflavone is 5 to 500 mL / g, more preferably the volume mass ratio of the single solvent to 7,8-dihydroxyflavone is 5 mL / g, and / or, in method 1, the method for precipitating the solid employs a solvent evaporation method to precipitate the solid. and / or, in method 2, the single solvent is water, diethyl ether, more preferably the single solvent is water, and the two-component solvent is a combination of an organic solvent and water, a combination of butyl formate and n-heptane, a combination of ethylene glycol methyl ether and chloroform, a combination of acetonitrile and toluene, more preferably the two-component solvent is a combination of an organic solvent and water. and / or, in method 2, the organic solvent is one of tetrahydrofuran, isopropyl alcohol, acetonitrile, methanol, ethanol, dimethyl sulfoxide, N,N-dimethylformamide, and ethylene glycol dimethyl ether, and more preferably the organic solvent is tetrahydrofuran. and / or, in method 2, the two-component solvent has a volume ratio of organic solvent to water of 1:5 to 1:10, a volume ratio of butyl formate to n-heptane of 1:1, a volume ratio of ethylene glycol methyl ether to chloroform of 1:5, a volume ratio of acetonitrile to toluene of 1:1, and more preferably, the two-component solvent has a volume ratio of organic solvent to water of 1:5, and / or, in method 3, the good solvent is one of N,N-dimethylformamide, tetrahydrofuran, and ethylene glycol dimethyl ether, and the antisolvent is one of water, toluene, methylene chloride, and diethyl ether, more preferably the good solvent is N,N-dimethylformamide and the antisolvent is water. and / or, in method 3, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10, and more preferably, the volume ratio of the good solvent to the antisolvent is 1:1. and / or, in method 4, the good solvent is one of N,N-dimethylformamide and isopropyl alcohol, and the antisolvent is one of chloroform, toluene and water, more preferably the good solvent is N,N-dimethylformamide and the antisolvent is chloroform. and / or, in method 4, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10, and more preferably, the volume ratio of the good solvent to the antisolvent is 1:1. and / or, in method 5, the antisolvent is water, and the good solvent is one of N,N-dimethylformamide, tetrahydrofuran, ethanol, and ethylene glycol methyl ether, more preferably the antisolvent is water, and the good solvent is N,N-dimethylformamide.

[0072] and / or, in method 5, the volume ratio of the antisolvent to the good solvent is 1:0.8 to 1:1, more preferably 1:0.8. In Method 1, the single solvent is one of isopropyl acetate and N,N-dimethylformamide, and more preferably the single solvent is N,N-dimethylformamide. and / or, in method 1, the temperature at which the solid is precipitated is room temperature. and / or, in method 2, the temperature of the suspension is 20 to 60°C, more preferably 25°C. and / or, in method 3, the temperature of the dropping is 15 to 50°C, more preferably 25°C. and / or, in method 4, the temperature of the dropping is 15 to 50°C, more preferably 25°C. And / or, in method 5, the cooling temperature is -15 to 4°C.

[0073] This disclosure relates to a method for preparing 7,8-dihydroxyflavone having the characteristics of at least one crystalline form N, in particular pure crystalline form N. The method may be, for example, as follows:

[0074] In Method 1, the single solvent is methanol. and / or, in method 1, the volume mass ratio of the single solvent to 7,8-dihydroxyflavone is 5 to 500 mL / g, more preferably 5 mL / g. and / or, in method 1, the method for precipitating the solid employs a solvent evaporation method to precipitate the solid. and / or, in method 2, the single solvent is methanol, and the two-component solvent is a combination of methanol and water. and / or, in method 2, the two-component solvent has a volume ratio of methanol to water of 1:5 to 1:10, and more preferably, the two-component solvent has a volume ratio of methanol to water of 1:5. and / or, in method 3, the good solvent is ethanol, and the antisolvent is one of methyl-t-butyl ether and toluene, more preferably the antisolvent is methyl-t-butyl ether. and / or, in method 3, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10, and more preferably, the volume ratio of the good solvent to the antisolvent is 1:1. and / or, in method 4, the good solvent is ethanol, and the antisolvent is one of cyclohexane, n-heptane, and water, more preferably the antisolvent is cyclohexane. and / or, in method 4, the volume ratio of the good solvent to the antisolvent is 1:1 to 1:10, and more preferably, the volume ratio of the good solvent to the antisolvent is 1:1. and / or, in method 5, the antisolvent is one of toluene, isopropyl acetate, 4-methyl-2-pentanone, water, and acetonitrile, and the good solvent is one of ethanol and methanol, more preferably the antisolvent is toluene and the good solvent is ethanol. and / or, in method 5, the volume ratio of the antisolvent to the good solvent is 1:0.8 to 1:1, more preferably 1:0.8. and / or, in method 1, the single solvent is methanol, and / or, in method 1, the temperature at which the solid is precipitated is room temperature. and / or, in method 2, the suspension temperature is 20 to 60°C, more preferably 50°C. and / or, in method 3, the temperature of the dropping is 15 to 50°C, more preferably 25°C. and / or, in method 4, the temperature of the dropping is 15 to 50°C, more preferably 25°C. and / or, in method 5, the cooling temperature is -15 to 4°C, and more preferably, the cooling temperature is 0°C.

[0075] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form A, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 9.1±0.2°, 17.2±0.2°, 20.5±0.2°, and 27.3±0.2°. Crystalline form A is a hydrate. The X-ray diffraction pattern of crystalline form A is shown in Figure 19. The positions of the characteristic X-ray diffraction peaks of crystalline form A, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0076] On the one hand, when crystal form A is sufficiently dried, it transforms into crystal form N and cannot exist for a long time under dry conditions. On the other hand, when crystal form A is heated to 130°C, it transforms into a mixed crystal having crystal form F, and when heated to 160°C, it transforms into a mixed crystal having crystal form F and crystal form J.

[0077] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form B, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 6.4±0.2°, 9.4±0.2°, 10.9±0.2°, 12.8±0.2°, 13.9±0.2°, 17.2±0.2°, 18.8±0.2°, 20.4±0.2°, and 20.7±0.2°. Crystalline form B is an anhydrous form containing a small amount of adsorbent solvent. The X-ray diffraction pattern of crystalline form B is shown in Figure 20. The positions of the characteristic X-ray diffraction peaks of crystalline form B, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0078] On the one hand, crystal form B cannot exist at room temperature for a long time because it is converted to crystal form C at room temperature and humidity. On the other hand, when crystal form B is further heated to 110°C, a characteristic peak of a small amount of crystal form J appears, and at 180°C it is converted to a mixed crystal having crystal forms F and J.

[0079] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form E, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 14.2±0.2°, 16.3±0.2°, 20.3±0.2°, 23.3±0.2°, 24.2±0.2°, and 27.3±0.2°. Crystalline form E is either a hydrate or a dimethyl sulfoxide solvate. The X-ray diffraction pattern of crystalline form E is shown in Figure 21. The positions of the characteristic X-ray diffraction peaks of crystalline form E, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0080] Crystal form E is converted to crystal form N at room temperature.

[0081] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form F, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 6.0±0.2°, 15.9±0.2°, 17.2±0.2°, 19.2±0.2°, and 26.9±0.2°. Crystalline form F is an anhydrous substance containing a small amount of adsorbent solvent. The X-ray diffraction pattern of crystalline form F is shown in Figure 22. The positions of the characteristic X-ray diffraction peaks of crystalline form F, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0082] On the one hand, crystal form F cannot exist for a long period of time at room temperature because it is converted to crystal form B at room temperature. On the other hand, when crystal form F is heated to 220°C, it is converted to a mixed crystal of crystal forms J and P.

[0083] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form G, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 11.6±0.2°, 18.5±0.2°, and 35.4±0.2°. Crystalline form G is a hydrate. The X-ray diffraction pattern of crystalline form G is shown in Figure 23. The positions of the characteristic X-ray diffraction peaks of crystalline form G, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0084] When crystalline form G is heated to 140°C, it transforms into crystalline form T, which contains a small amount of crystalline form J.

[0085] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form H, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 8.3±0.2°, 11.2±0.2°, 11.6±0.2°, 16.7±0.2°, and 21.5±0.2°. The X-ray diffraction pattern of crystalline form H is shown in Figure 24. The positions of the characteristic X-ray diffraction peaks of crystalline form H, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0086] Crystalline form H can transform into other crystalline forms at room temperature and cannot exist at room temperature for extended periods.

[0087] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form I, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 10.5±0.2°, 17.0±0.2°, and 20.5±0.2°. The X-ray diffraction pattern of crystalline form I is shown in Figure 25. The positions of the characteristic X-ray diffraction peaks of crystalline form I, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0088] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form J, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 4.3±0.2°, 5.7±0.2°, 6.8±0.2°, 8.3±0.2°, 12.4±0.2°, 12.8±0.2°, 17.2±0.2°, and 18.0±0.2°. Crystalline form J is anhydrous. The X-ray diffraction pattern of crystalline form J is shown in Figure 26. The positions of the characteristic X-ray diffraction peaks of crystalline form J, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0089] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form K, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 10.8±0.2°, 19.5±0.2°, 22.1±0.2°, 22.4±0.2°, 24.7±0.2°, and 28.2±0.2°. Crystalline form K is a 1:1 dioxane solvate. The X-ray diffraction pattern of crystalline form K is shown in Figure 27. The positions of the characteristic X-ray diffraction peaks of crystalline form K, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0090] When crystal form K is heated to 150°C and then cooled to room temperature, it transforms into a mixed crystal of crystal forms F and J.

[0091] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form L, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 10.6±0.2°, 11.3±0.2°, 12.0±0.2°, 15.5±0.2°, 17.7±0.2°, 19.5±0.2°, and 22.6±0.2°. The X-ray diffraction pattern of crystalline form L is shown in Figure 28. The positions of the characteristic X-ray diffraction peaks of crystalline form L, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0092] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form M, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 8.5±0.2°, 14.8±0.2°, 16.5±0.2°, and 19.0±0.2°. Crystalline form M is an acetone solvate. The X-ray diffraction pattern of crystalline form M is shown in Figure 29. The positions of the characteristic X-ray diffraction peaks of crystalline form M, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0093] Crystalline form M transforms into crystalline form C at room temperature and cannot exist at room temperature for extended periods.

[0094] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form O, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 3.4±0.2°, 5.9±0.2°, 8.9±0.2°, 10.9±0.2°, 11.7±0.2°, 13.6±0.2°, 14.1±0.2°, 15.2±0.2°, 20.4±0.2°, and 20.7±0.2°. Crystalline form O is an anhydrous or chloroform solvate with a small amount of solvent residue. The X-ray diffraction pattern of crystalline form O is shown in Figure 30. The positions of the characteristic X-ray diffraction peaks of crystalline form O, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0095] Crystalline form O converts to crystalline form C at room temperature and cannot exist at room temperature for extended periods.

[0096] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form Q, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 3.6±0.2°, 7.7±0.2°, 12.1±0.2°, 13.0±0.2°, 16.6±0.2°, and 20.2±0.2°. The X-ray diffraction pattern of crystalline form Q is shown in Figure 31. The positions of the characteristic X-ray diffraction peaks of crystalline form Q, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0097] Crystalline form Q transforms into crystalline form C at room temperature and cannot exist at room temperature for extended periods.

[0098] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form R, characterized by its X-ray diffraction pattern having an X-ray diffraction peak at approximately 2θ. The X-ray diffraction pattern of crystalline form R is shown in Figure 32. The position of the characteristic X-ray diffraction peak of crystalline form R, along with the position of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, is shown in Table 1.

[0099] Crystal form R can only be obtained by heating crystal form N to 100°C, and it cannot exist at room temperature for extended periods.

[0100] In one comparative embodiment, the disclosure provides a 7,8-dihydroxyflavone having at least one crystalline form S, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 8.6±0.2°, 17.3±0.2°, and 22.6±0.2°. The X-ray diffraction pattern of crystalline form S is shown in Figure 33. The positions of the characteristic X-ray diffraction peaks of crystalline form S, along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone, are shown in Table 1.

[0101] Crystalline form S converts to crystalline form C at room temperature and cannot exist at room temperature for extended periods.

[0102] In one comparative embodiment, the disclosure further provides a 7,8-dihydroxyflavone having at least one crystalline form T, characterized by its X-ray diffraction pattern with X-ray diffraction peaks at 2θ values ​​of approximately 9.3±0.2°, 13.0±0.2°, 15.4±0.2°, and 23.8±0.2°. The X-ray diffraction pattern of crystalline form T is shown in Figure 34. The positions of the characteristic X-ray diffraction peaks of crystalline form T are shown in Table 1 along with the positions of the characteristic X-ray diffraction peaks of other crystalline forms of 7,8-dihydroxyflavone.

[0103] Crystalline form T can only be obtained by heating crystalline form G to 140°C, and it cannot exist at room temperature for extended periods.

[0104] [Table 1]

[0105] In one embodiment, the disclosure provides a pharmaceutical composition comprising a crystalline form P of 7,8-dihydroxyflavone and at least one pharmaceutically acceptable excipient. Preferably, the pharmaceutical composition is in the form of an oral solid dosage form.

[0106] In one embodiment, tablets are prepared using a standard formulation method, and it is found that the crystalline form P within the tablets can remain stable for 6 months under harsh conditions such as light irradiation, high temperature, and high humidity. The stability of the crystalline form P of 7,8-dihydroxyflavone within the tablets is not limited by the specific dosage form.

[0107] In one embodiment, tablets are prepared using a standard formulation method, and it is discovered that the crystalline form P within the tablets has better dissolution.

[0108] In one embodiment, the disclosure further provides a pharmaceutical composition comprising crystalline form C of 7,8-dihydroxyflavone and at least one pharmaceutically acceptable excipient. Preferably, the pharmaceutical composition is in the form of an oral solid dosage form.

[0109] In one embodiment, tablets are prepared using a standard formulation method, and it is found that crystalline form C within the tablets can remain stable for 6 months under harsh conditions such as light irradiation, high temperature, and high humidity. The stability of crystalline form C of 7,8-dihydroxyflavone within the tablets is not limited by the specific dosage form.

[0110] In one embodiment, the present disclosure further provides a pharmaceutical composition comprising crystalline form N of 7,8-dihydroxyflavone and at least one pharmaceutically acceptable excipient. Preferably, the pharmaceutical composition is in the form of an oral solid dosage form.

[0111] In one embodiment, the pharmaceutical composition according to the Disclosure comprises one or more crystalline polymorphs of 7,8-dihydroxyflavone described in the Invention. In addition to the active ingredient, the pharmaceutical composition according to the Disclosure may have one or more excipients. Different purposes can be achieved by adding various excipients to the composition.

[0112] Fillers can increase the volume of a solid medicinal composition, making it easier for patients and caregivers to use the medicinal dosage form containing the composition. Fillers used in solid compositions include, for example, microcrystalline cellulose, ultrafine cellulose, lactose, starch, pregelatinized starch, calcium carbonate, calcium sulfate, sugars, glucose binders, dextrin, glucose, calcium hydrogen phosphate dihydrate, calcium phosphate, kaolin, magnesium carbonate, magnesium oxide, maltodextrin, mannitol, polymethacrylate, potassium chloride, powdered cellulose, sodium chloride, sorbitol, and talc powder.

[0113] Solid pharmaceutical compositions pressed into dosage forms such as tablets may contain excipients, the function of which includes assisting the binding of the active ingredient with other excipients after pressing. Binders for solid pharmaceutical compositions include povidone, gum arabic, alginic acid, carbomer, sodium carboxymethylcellulose, dextrin, ethylcellulose, gelatin, guar gum, hydrogenated vegetable oil, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, liquid glucose, magnesium aluminum silicate, maltodextrin, methylcellulose, polymethacrylate, polyvinylpyrrolidone, pregelatinized starch, sodium alginate, and starch.

[0114] By adding disintegrants, the dissolution rate of pressed solid medicinal compositions in the patient's stomach can be increased. Disintegrants include sodium carboxymethyl starch, alginic acid, calcium carboxymethylcellulose, sodium carboxymethylcellulose, colloidal silica, cross-linked sodium carboxymethylcellulose, cross-linked polyvinylpyrrolidone, guar gum, magnesium aluminum silicate, methylcellulose, microcrystalline cellulose, potassium polariphosphate, powdered cellulose, pregelatinized starch, sodium alginate, sodium starch glycolate, and starch.

[0115] When a powdered composition is pressed into a tablet-like dosage form, the composition is subjected to pressure from a die and a punch. Certain excipients and active ingredients tend to adsorb to the surfaces of the punch and die, and such tendency can result in indentations and other surface irregularities in the product. By adding a lubricant to the composition, adsorption can be reduced and the release of the product from the die can be facilitated. Lubricants include magnesium stearate, calcium stearate, glycerin monostearate, glyceryl palmitostearate, hydrogenated castor oil, hydrogenated vegetable oil, mineral oil, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, talc powder, and zinc stearate.

[0116] The solid dosage form of the pharmaceutical composition relating to this disclosure may be, for example, a tablet, powder, capsule, suppository, pouch, tablet, liquid syrup, suspending agent, etc. In one embodiment, the dosage form of this disclosure is a tablet.

[0117] In one embodiment, the Disclosure further provides a method for treating diseases associated with abnormalities in the tyrosine kinase receptor B (TrkB) signaling pathway, such as central nervous system injury, peripheral nerve injury, neurodegenerative lesions, psychiatric disorders, hereditary neurological disorders, ophthalmic diseases, metabolic diseases, pain, cardiovascular diseases, tumors, or other related diseases, the method comprising administering to a subject the crystalline form of 7,8-dihydroxyflavone of the Disclosure and at least one pharmaceutically acceptable excipient. In one embodiment, the disease associated with abnormalities in the TrkB signaling pathway is a disease resulting from low expression or insufficient activation of TrkB. In the Art, it is known that 7,8-dihydroxyflavone can activate the TrkB downstream signaling pathway as a specific small molecule agonist of TrkB.

[0118] In one embodiment, the present disclosure provides a method for treating or preventing cognitive decline associated with traumatic brain injury or aging, the method comprising administering the crystalline form of the 7,8-dihydroxyflavone to a subject.

[0119] In one embodiment, the disclosure provides crystalline forms P, C, or N of 7,8-dihydroxyflavone for treating or preventing cognitive decline associated with traumatic brain injury or aging.

[0120] In one embodiment, the present disclosure provides the use of crystalline forms P, C, and / or N of 7,8-dihydroxyflavone in the preparation of agents for treating or preventing cognitive decline associated with traumatic brain injury or aging. Those skilled in the art know that the peak intensity and / or peak characteristics of X-ray powder diffraction may vary depending on experimental conditions. In addition, due to the different accuracies of instruments, the measured 2θ values ​​may have an error of about ±0.2 degrees. On the other hand, the relative intensity value of the peak may exhibit a deviation of about ±20% in the measured peak intensity because it depends more on certain properties of the measured sample than on the position of the peak, such as the size and purity of the crystal. Despite test errors, instrument errors, and orientation priorities, those skilled in the art can still obtain sufficient information from the X-ray powder diffraction data relating to the present disclosure to authenticate each crystal form.

[0121] In this disclosure, the expression "having at least one crystalline form "#"" (where "#" is an alphabet letter) means that 7,8-dihydroxyflavone has a characteristic X-ray peak of at least crystalline form "#".

[0122] In this disclosure, "solvent volatilization method" refers to a commonly used crystallization method in the field of crystalline forms, in which the solution is continuously volatilized to change from an unsaturated to a supersaturated state, thereby precipitating crystals.

[0123] The above preferred conditions can be combined in any way, provided that they do not contradict common sense in this field.

[0124] Unless otherwise specified, all reagents and raw materials used in this disclosure are commercially available. [Examples]

[0125] The present invention will be further described below in accordance with the methods of the examples, but the present invention is not limited to the scope of the above examples. Experimental methods in the following examples that do not specify concrete conditions are selected according to conventional methods and conditions or according to the product specification.

[0126] Test method Nuclear magnetic analysis (1H NMR) A small milligram of solid sample was dissolved in dimethyl sulfoxide-d6 solvent and subjected to nuclear magnetic analysis using a Bruker AVANCE NEO 400 (Bruker, GER).

[0127] X-ray powder diffraction (XRPD) Solid samples obtained from the experiment were analyzed using a Bruker D8 Advance X-ray powder diffractometer (Bruker, GER). The 2θ scanning angle ranged from 3° to 45°, the scanning step was 0.02°, and the exposure time was 0.08 seconds. When testing the samples, the tube voltage and current were 40kV and 40mA, respectively, and a zero-background sample plate was used.

[0128] Thermogravimetric analysis (TGA) The thermogravimetric analyzer was a TA Discovery 55 (TA, US). 2-5 mg samples were placed on a balanced, open aluminum sample plate and automatically weighed in a TGA heating furnace. The samples were heated to the final temperature at a rate of 10°C / min, with a nitrogen purge rate of 60 mL / min on the sample and 40 mL / min on the balance.

[0129] Differential Scanning Calorimetry (DSC) The differential scanning calorimetry analyzer was a TA Discovery 2500 (TA, US). After accurately weighing 1-2 mg of sample, it was placed on a perforated DSC Tzero sample plate and heated to the final temperature at a rate of 10°C / min, with a nitrogen purge rate of 50 mL / min in the furnace.

[0130] Dynamic water adsorption analysis (DVS) Dynamic moisture adsorption analysis was performed using DVS Intrinsic (SMS, UK). The test employed a gradient mode, with humidity changes of 50%-95%-0%-50%. Within the range of 0% to 90%, the humidity change for each gradient was 10%, and the gradient endpoint was determined using the dm / dt method, where dm / dt remained below 0.002% for 10 minutes. After the test, XRPD analysis was performed on the sample to confirm whether or not the solid morphology had changed. The criteria for judgment are shown in Table 2.

[0131] [Table 2]

[0132] Polarized light microscopy (PLM) The polarizing microscope model was Nikon Ci-POL (Nikon, JPN). A small sample was placed on a glass slide, and an appropriate lens was selected to observe the sample's morphology.

[0133] Example 1: Preparation of crystalline form P of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 1.0 mL of ethanol and 1.0 mL of ethyl acetate until a suspension was formed, and the suspension was stirred at 50°C for 7 days. After that, the suspension was centrifuged, and the solid was vacuum-dried at room temperature.

[0134] XRPD detection revealed that the X-ray powder diffraction, expressed in 2θ angles, was 4.1±0.2°, 6.2±0.2°, 7.0±0.2°, 7.3±0.2°, 7.9±0.2°, 8.3±0.2°, 11.0±0.2°, 12.5±0.2°, 13.6±0.2°, 14.0±0.2°, 15.8±0.2°, 16.6±0.2°, 17.2±0.2°, 18.4±0.2°, 1 Diffraction peaks were observed at 8.7±0.2°, 19.4±0.2°, 20.8±0.2°, 21.4±0.2°, 22.8±0.2°, 24.1±0.2°, 25.0±0.2°, 25.9±0.2°, 27.0±0.2°, 27.4±0.2°, 31.2±0.2°, 31.4±0.2°, and 33.7±0.2°, and their XRPD spectra are shown in Figure 1.

[0135] ¹H NMR detection revealed no clear solvent residue peaks, indicating that the crystalline form P of 7,8-dihydroxyflavone is not a solvate. Its ¹H NMR spectrum is shown in Figure 12.

[0136] TGA detection revealed no significant weight loss in the crystalline form P of 7,8-dihydroxyflavone, and its TGA spectrum is shown in Figure 4.

[0137] DSC detection revealed that crystalline form P of 7,8-dihydroxyflavone showed no endothermic signal corresponding to weight loss, and its DSC spectrum is shown in Figure 4.

[0138] DVS detection revealed that the dynamic water adsorption graph of crystalline form P of 7,8-dihydroxyflavone showed a weight increase of less than 0.1% within the relative humidity range of 0% to 95%, and its DVS spectrum is shown in Figure 5.

[0139] PLM detection revealed that the crystalline form P of 7,8-dihydroxyflavone was fine particles, with a particle size generally less than 20 μm, and its PLM spectrum is shown in Figure 11.

[0140] Example 2 Preparation of crystalline form C of 7,8-dihydroxyflavone Approximately 20 mg of the raw material was weighed and placed in an EP tube. A fixed amount of solvent was added sequentially at room temperature (approximately 25°C), and the solution was stirred and sonicated until the solid was completely dissolved. The resulting clear solution was left to stand open at room temperature until the solvent had completely evaporated or most of it had evaporated and a solid was obtained.

[0141] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 5.6±0.2°, 9.5±0.2°, 11.1±0.2°, 12.7±0.2°, 13.5±0.2°, 13.7±0.2°, 15.5±0.2°, 16.0±0.2°, 16.3±0.2°, 16.6±0.2°, 16.9±0.2°, and 17.4±0.2°. , 18.3±0.2°, 18.9±0.2°, 19.2±0.2°, 19.4±0.2°, 20.4±0.2°, 20.8±0.2°, 21.0±0.2°, 21.9±0.2°, 22.2±0.2°, 23.2±0.2°, 23.4±0.2°, 24.1±0.2°, 24.3±0.2°, 25.0±0.2°, 25.2 ±0.2°, 25.5±0.2°, 25.8±0.2°, 27.0±0.2°, 27.3±0.2°, 27.6±0.2°, 27.9±0.2°, 28.3±0.2°, 28.6±0.2°, 28.9±0.2°, 29.7±0.2°, 30.4±0.2°, 30.7±0.2°, 31.7±0.2°, 33.2±0.2° Diffraction peaks were observed at 33.6±0.2°, 34.3±0.2°, 34.5±0.2°, 34.6±0.2°, 35.1±0.2°, 35.7±0.2°, 36.3±0.2°, 37.7±0.2°, 39.8±0.2°, 42.6±0.2°, and 44.5±0.2°, and their XRPD spectra are shown in Figure 2.

[0142] ¹H NMR detection revealed no clear solvent residue peaks, indicating that the crystalline form C of 7,8-dihydroxyflavone is not a solvate. Its ¹H NMR spectrum is shown in Figure 15.

[0143] TGA detection revealed that the mass loss of crystalline form C of 7,8-dihydroxyflavone at 150°C was 3.7% of the mass before the loss. This weight loss is due to the weight of water within the hydrate crystal, and its TGA spectrum is shown in Figure 6.

[0144] DSC detection revealed that crystalline form C of 7,8-dihydroxyflavone showed an endothermic signal corresponding to weight loss between 100°C and 160°C, and a melting endothermic peak at 247°C. Its DSC spectrum is shown in Figure 6.

[0145] DVS detection revealed that the dynamic water adsorption graph of crystalline form C of 7,8-dihydroxyflavone showed a weight increase of less than 0.1% within the relative humidity range of 0% to 95%, and its DVS spectrum is shown in Figure 7.

[0146] PLM detection revealed that crystalline form C of 7,8-dihydroxyflavone was fine particles, with a particle size generally less than 5 μm, and its PLM spectrum is shown in Figure 14.

[0147] Example 3 Preparation of crystalline form N of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 1.0 mL of methanol until a suspension was formed, and the suspension was stirred at 50°C for 7 days. After that, the suspension was centrifuged, and the solid was vacuum-dried at room temperature.

[0148] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 5.7±0.2°, 9.3±0.2°, 11.1±0.2°, 11.4±0.2°, 11.9±0.2°, 12.3±0.2°, 13.1±0.2°, 14.2±0.2°, 14.9±0.2°, 15.9±0.2°, 16.8±0.2°, 17.1±0.2°, 17.7 ±0.2°, 18.7±0.2°, 18.9±0.2°, 19.2±0.2°, 20.2±0.2°, 20.7±0.2°, 21.1±0.2°, 22.1±0.2°, 22.4±0.2°, 23.1±0.2°, 23.2±0.2°, 23.4±0.2°, 23.8±0.2°, 24.6±0.2°, 25.2±0.2°, 25.7 ±0.2°, 26.3±0.2°, 26.6±0.2°, 26.9±0.2°, 27.3±0.2°, 28.9±0.2°, 28.0±0.2°, 29.0±0.2°, 29.4±0.2°, 29.6±0.2°, 30.4±0.2°, 31.3±0.2°, 31.9±0.2°, 32.1±0.2°, 32.4±0.2°, 35.0 Diffraction peaks were observed at ±0.2°, 35.4±0.2°, 36.2±0.2°, 36.8±0.2°, 38.2±0.2°, 38.4±0.2°, 38.6±0.2°, 38.9±0.2°, 41.0±0.2°, 41.4±0.2°, 42.0±0.2°, and 43.9±0.2°, and their XRPD spectra are shown in Figure 3.

[0149] ¹H NMR detection revealed no clear solvent residue peaks, indicating that the crystalline form N of 7,8-dihydroxyflavone is not a solvate. Its ¹H NMR spectrum is shown in Figure 18.

[0150] TGA detection revealed that the mass loss of crystalline form N of 7,8-dihydroxyflavone between 40°C and 80°C was 6.3% of the mass before the loss. This weight loss is due to the weight of water within the hydrate crystal, and its TGA spectrum is shown in Figure 8.

[0151] DSC detection revealed that crystalline form N of 7,8-dihydroxyflavone showed an endothermic peak corresponding to weight loss at 100°C, and heat dissipation signals at 136°C and 169°C, respectively. Its DSC spectrum is shown in Figure 8.

[0152] DVS detection revealed that the dynamic water adsorption graph of crystalline form N of 7,8-dihydroxyflavone showed a weight increase of less than 0.1% within the relative humidity range of 0% to 95%, and its DVS spectrum is shown in Figure 9.

[0153] PLM detection revealed that the crystalline form N of 7,8-dihydroxyflavone was fine particles, with a particle size generally less than 20 μm, and its PLM spectrum is shown in Figure 17.

[0154] Comparative Example 1: Preparation of crystalline form A of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 2.0 mL of ethanol until a suspension was formed, and the suspension was stirred at room temperature for 7 days. The suspension was then centrifuged, and the solid was vacuum-dried at room temperature.

[0155] XRPD detection revealed diffraction peaks in the X-ray powder diffraction, expressed as a 2θ angle, at 5.6±0.2°, 9.1±0.2°, 11.7±0.2°, 12.0±0.2°, 12.2±0.2°, 16.4±0.2°, 17.2±0.2°, 18.2±0.2°, 18.5±0.2°, 20.4±0.2°, 20.5±0.2°, 21.0±0.2°, 23.5±0.2°, 26.1±0.2°, 27.3±0.2°, 27.8±0.2°, 35.3±0.2°, and 41.1±0.2°. The XRPD spectra are shown in Figure 19.

[0156] Comparative Example 2: Preparation of crystalline form B of 7,8-dihydroxyflavone 40.0 mg of 7,8-dihydroxyflavone was added to 9.0 mL of ethyl formate until a suspension was formed, and the suspension was stirred at room temperature for 7 days. The suspension was then centrifuged, and the solid was vacuum-dried at room temperature.

[0157] XRPD detection revealed the following X-ray powder diffraction values, expressed in 2θ angles: 3.1±0.2°, 3.4±0.2°, 3.7±0.2°, 5.5±0.2°, 5.9±0.2°, 6.4±0.2°, 7.3±0.2°, 8.2±0.2°, 8.5±0.2°, 9.4±0.2°, 9.8±0.2°, and 10.3±0. 2°, 10.9±0.2°, 11.2±0.2°, 11.8±0.2°, 12.8±0.2°, 13.4±0.2°, 13.9±0.2°, 14.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.7±0.2°, 16.3±0.2°, 16.7±0.2°, 17.2±0.2°, 17.9±0.2°, 18.8±0.2°, 19.3±0.2°, 19.8±0.2°, 20.4±0.2°, 20.7±0.2°, 22.1±0.2°, 22.2±0.2°, 22.5±0.2°, 23.7±0.2°, 24.2±0.2°, 24.9±0.2°, 25.3±0.2°, 26. Diffraction peaks were observed at 0±0.2°, 26.7±0.2°, 27.8±0.2°, 28.1±0.2°, 28.3±0.2°, 29.0±0.2°, 30.2±0.2°, 32.5±0.2°, 34.7±0.2°, and 41.46±0.2°, and their XRPD spectra are shown in Figure 20.

[0158] Comparative Example 3: Preparation of crystalline form E of 7,8-dihydroxyflavone Approximately 20 mg of the raw material was weighed and placed in an EP tube. 2 mL of ethylene glycol dimethyl ether was added at room temperature (approximately 25°C), and the solution was stirred and sonicated until the solid was completely dissolved. The resulting clear solution was left to stand open at room temperature until the solvent had completely or mostly evaporated, yielding a solid.

[0159] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 5.6±0.2°, 8.9±0.2°, 11.1±0.2°, 11.6±0.2°, 11.9±0.2°, 12.1±0.2°, 13.5±0.2°, 14.0±0.2°, 14.2±0.2°, 15.8±0.2°, 16.3±0.2°, 16.6±0.2°, 17 0.0±0.2°, 17.3±0.2°, 17.9±0.2°, 18.3±0.2°, 18.6±0.2°, 18.8±0.2°, 19.0±0.2°, 19.3±0.2°, 20.3±0.2°, 21.2±0.2°, 21.7±0.2°, 22.0±0.2°, 22.3±0.2°, 22.6±0.2°, 23.3±0.2°, 23.8±0.2°, 24.2±0.2°, 25.0±0.2°, 25.4±0.2°, 25.8±0.2°, 26.5±0.2°, 26.9±0.2°, 27.3±0.2°, 27.8±0.2°, 28.1±0.2°, 28.6±0.2°, 29.2±0.2°, 29.7±0.2°, 30.7±0.2°, 31.8±0. Diffraction peaks were observed at 2°, 31.9±0.2°, 34.0±0.2°, 34.8±0.2°, 34.9±0.2°, 35.5±0.2°, 36.2±0.2°, 37.6±0.2°, 39.5±0.2°, 40.6±0.2°, 42.6±0.2°, and 43.2±0.2°, and their XRPD spectra are shown in Figure 21.

[0160] Comparative Example 4: Preparation of crystalline form F of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 10.0 mL of acetonitrile until a suspension was formed, and the suspension was stirred at room temperature for 7 days. After that, the suspension was centrifuged, and the solid was vacuum-dried at room temperature.

[0161] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 6.0±0.2°, 10.3±0.2°, 12.0±0.2°, 12.3±0.2°, 13.7±0.2°, 15.0±0.2°, 15.9±0.2°, 17.2±0.2°, 18.2±0.2°, 19.2±0.2°, 20.9±0.2°, 21.9±0.2°, 22.7±0.2°, 24.2±0.2°, 24.7±0.2°, and 25.5±0.2°. Diffraction peaks were observed at 26.2±0.2°, 26.9±0.2°, 27.6±0.2°, 28.2±0.2°, 29.5±0.2°, 30.1±0.2°, 31.0±0.2°, 32.0±0.2°, 33.3±0.2°, 33.7±0.2°, 35.5±0.2°, 36.4±0.2°, 36.9±0.2°, 38.0±0.2°, and 38.5±0.2°, and their XRPD spectra are shown in Figure 22.

[0162] Comparative Example 5: Preparation of crystalline form G of 7,8-dihydroxyflavone 160 mg of 7,8-dihydroxyflavone was weighed, and 11 mL of ethylene glycol dimethyl ether was added dropwise at room temperature to completely dissolve the sample. 1.3 mL of the solution was taken and added dropwise to 6.5 mL of water. After stirring for 15 min to 1 hour, the precipitated solid was centrifuged, and then the solid was vacuum-dried at room temperature.

[0163] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 11.6±0.2°, 12.1±0.2°, 13.1±0.2°, 13.5±0.2°, 16.8±0.2°, 18.5±0.2°, 20.3±0.2°, 21.3±0.2°, 21.7±0.2°, 22.9±0.2°, 23.4±0.2°, 24.2°. Diffraction peaks were observed at ±0.2°, 24.7±0.2°, 26.5±0.2°, 26.8±0.2°, 28.2±0.2°, 30.0±0.2°, 31.9±0.2°, 34.2±0.2°, 35.4±0.2°, 38.9±0.2°, and 42.0±0.2°, and their XRPD spectra are shown in Figure 23.

[0164] Comparative Example 6: Preparation of crystalline form H of 7,8-dihydroxyflavone 160 mg of 7,8-dihydroxyflavone was weighed, and 11 mL of ethylene glycol dimethyl ether was added dropwise at room temperature to completely dissolve the sample. 1.3 mL of the solution was taken, and 1.0 mL of n-heptane was added dropwise. After stirring for 15 min to 1 hour at room temperature, the precipitated solid was centrifuged, and then the solid was vacuum-dried at room temperature.

[0165] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 7.2±0.2°, 8.3±0.2°, 8.8±0.2°, 9.5±0.2°, 10.2±0.2°, 11.2±0.2°, 11.6±0.2°, 12.2±0.2°, 12.4±0.2°, 13.4±0.2°, 13.7±0.2°, 14.5±0.2°, 14.9±0.2°, 15.5±0.2°, 16.2±0.2°, 16.7±0.2°, 18.4±0.2°. Diffraction peaks were observed at 19.1±0.2°, 19.2±0.2°, 19.9±0.2°, 20.3±0.2°, 20.9±0.2°, 21.5±0.2°, 21.9±0.2°, 23.1±0.2°, 23.9±0.2°, 25.5±0.2°, 25.8±0.2°, 27.6±0.2°, 27.7±0.2°, 28.0±0.2°, 28.9±0.2°, and 31.0±0.2°, and their XRPD spectra are shown in Figure 24.

[0166] Comparative Example 7: Preparation of crystalline form I of 7,8-dihydroxyflavone 160 mg of 7,8-dihydroxyflavone was weighed out, and 11 mL of ethylene glycol dimethyl ether was added dropwise at room temperature to completely dissolve the sample. 1.3 mL of the solution was taken, and 5.0 mL of water was added dropwise. After stirring at room temperature for 15 min to 1 hour, the precipitated solid was centrifuged, and then the solid was vacuum-dried at room temperature.

[0167] XRPD detection revealed diffraction peaks in the X-ray powder diffraction spectrum, expressed as a 2θ angle, at 7.0±0.2°, 8.3±0.2°, 10.5±0.2°, 12.3±0.2°, 13.0±0.2°, 14.0±0.2°, 14.8±0.2°, 16.7±0.2°, 17.0±0.2°, 20.5±0.2°, 21.8±0.2°, 22.0±0.2°, 22.1±0.2°, 23.8±0.2°, 25.9±0.2°, 27.1±0.2°, 27.6±0.2°, and 33.7±0.2°. The XRPD spectra are shown in Figure 25.

[0168] Comparative Example 8: Preparation of crystalline form J of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 10.0 mL of toluene until a suspension was formed, and the suspension was stirred at room temperature for 7 days. After that, the suspension was centrifuged, and the solid was vacuum-dried at room temperature.

[0169] XRPD detection revealed that the X-ray powder diffraction, expressed in 2θ angles, was 4.3±0.2°, 5.7±0.2°, 6.8±0.2°, 8.3±0.2°, 10.6±0.2°, 10.8±0.2°, 11.4±0.2°, 12.4±0.2°, 12.8±0.2°, 13.9±0.2°, 15.1±0.2°, 16.0±0.2°, 16.7±0.2°, 17.1±0.2°, 18.0±0.2°, 1 Diffraction peaks were observed at 9.2±0.2°, 20.1±0.2°, 20.5±0.2°, 21.2±0.2°, 21.7±0.2°, 22.9±0.2°, 24.0±0.2°, 24.2±0.2°, 25.1±0.2°, 26.3±0.2°, 26.9±0.2°, 28.0±0.2°, 28.6±0.2°, and 32.1±0.2°, and their XRPD spectra are shown in Figure 26.

[0170] Comparative Example 9: Preparation of crystalline form K of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 2.0 mL of dioxane until a suspension was formed, and the suspension was stirred at room temperature for 7 days. The suspension was then centrifuged, and the solid was vacuum-dried at room temperature.

[0171] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 9.9±0.2°, 10.8±0.2°, 12.0±0.2°, 12.4±0.2°, 13.4±0.2°, 14.3±0.2°, 14.7±0.2°, 15.7±0.2°, 17.5±0.2°, 17.9±0.2°, 19.5±0.2°, 20.4±0.2°, 20.7±0.2°, 20.9±0.2°, 22.1±0.2°, 22.4±0.2°, 23.4±0.2°, 23.9±0.2°, 24.7±0.2°, 25.8±0.2°, 26.3±0.2°, 27.3±0.2°, 27.4 Diffraction peaks were observed at ±0.2°, 28.1±0.2°, 28.7±0.2°, 29.6±0.2°, 30.4±0.2°, 31.0±0.2°, 31.4±0.2°, 31.9±0.2°, 32.4±0.2°, 33.1±0.2°, 33.7±0.2°, 34.2±0.2°, 34.8±0.2°, 35.4±0.2°, 36.2±0.2°, 37.0±0.2°, 37.8±0.2°, 38.8±0.2°, 39.6±0.2°, 40.9±0.2°, 43.8±0.2°, and 44.8±0.2°, and their XRPD spectra are shown in Figure 27.

[0172] Comparative Example 10: Preparation of crystalline form L of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 10.0 mL of methyl formate until a suspension was formed, and the suspension was stirred at room temperature for 7 days. After that, the suspension was centrifuged, and the solid was vacuum-dried at room temperature.

[0173] XRPD detection revealed the following X-ray powder diffraction values, expressed in 2θ angles: 7.7±0.2°, 10.6±0.2°, 11.3±0.2°, 12.0±0.2°, 12.4±0.2°, 15.5±0.2°, 16.9±0.2°, 17.7±0.2°, 18.9±0.2°, 19.5±0.2°, 20.0±0.2°, 21.0±0.2°, 21.3±0.2°, 22.0±0.2°, 22.2±0.2°, 22.6±0.2°, and 23.3±0.2°. Diffraction peaks were observed at 23.7±0.2°, 25.5±0.2°, 25.6±0.2°, 26.3±0.2°, 27.3±0.2°, 27.8±0.2°, 28.9±0.2°, 29.3±0.2°, 30.4±0.2°, 31.6±0.2°, 32.1±0.2°, 32.3±0.2°, 32.4±0.2°, 34.2±0.2°, 34.9±0.2°, and 37.9±0.2°, and their XRPD spectra are shown in Figure 28.

[0174] Comparative Example 11: Preparation of crystalline form M of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 2.0 mL of acetone until a suspension was formed, and the suspension was stirred at room temperature for 7 days. The suspension was then centrifuged, and the solid was vacuum-dried at room temperature.

[0175] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 8.5±0.2°, 11.1±0.2°, 14.5±0.2°, 14.8±0.2°, 16.5±0.2°, 18.1±0.2°, 18.3±0.2°, 19.0±0.2°, 20.8±0.2°, 22.0±0.2°, 22.3±0.2°, 22.8±0.2°, and 14.2±0.2°. Diffraction peaks were observed at 24.4±0.2°, 25.4±0.2°, 25.9±0.2°, 26.4±0.2°, 26.6±0.2°, 27.7±0.2°, 28.3±0.2°, 29.0±0.2°, 30.5±0.2°, 30.9±0.2°, 32.5±0.2°, and 33.3±0.2°, and their XRPD spectra are shown in Figure 29.

[0176] Comparative Example 12: Preparation of crystalline form O of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 1.0 mL of chloroform until a suspension was formed, and the suspension was stirred at 50°C for 7 days. After that, the suspension was centrifuged, and the solid was vacuum-dried at room temperature.

[0177] XRPD detection revealed that the X-ray powder diffraction, expressed in 2θ angles, was 3.4±0.2°, 5.9±0.2°, 7.0±0.2°, 8.5±0.2°, 8.9±0.2°, 10.3±0.2°, 10.9±0.2°, 11.7±0.2°, 12.8±0.2°, 13.6±0.2°, 14.1±0.2°, 14.7±0.2°, 15.2±0.2°, 16.0±0.2°, 17.0±0.2°, 17.3±0.2°, 17.9±0.2°, 18.6±0.2°, 19.3±0.2°, 19.8±0.2°, 20.4±0.2°, 2 Diffraction peaks were observed at 0.7±0.2°, 22.1±0.2°, 22.6±0.2°, 23.9±0.2°, 25.1±0.2°, 25.5±0.2°, 25.7±0.2°, 27.1±0.2°, 27.3±0.2°, 27.5±0.2°, 28.3±0.2°, 28.9±0.2°, 29.5±0.2°, 29.7±0.2°, 29.8±0.2°, 32.6±0.2°, 32.7±0.2°, 36.2±0.2°, 41.3±0.2°, and 41.4±0.2°, and their XRPD spectra are shown in Figure 30.

[0178] Comparative Example 13: Preparation of crystalline form Q of 7,8-dihydroxyflavone 20.0 mg of 7,8-dihydroxyflavone was added to 0.2 mL of dioxane and 1.0 mL of chloroform until a suspension was formed, and the suspension was stirred at 50°C for 7 days. After that, the suspension was centrifuged, and the solid was vacuum-dried at room temperature.

[0179] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 3.2±0.2°, 3.6±0.2°, 4.5±0.2°, 5.6±0.2°, 6.1±0.2°, 7.3±0.2°, 7.7±0.2°, 8.8±0.2°, 9.4±0.2°, 9.7±0.2°, 10.9±0.2°, 11.2±0.2°, 12.1±0.2°, 13.0±0.2°, 13.9±0.2°, 14.5±0.2°, 15.3±0.2°, 15.5±0.2°, 16.2±0.2°, 16.6±0.2°, 17.3±0.2°, 17.7 Diffraction peaks were observed at ±0.2°, 18.1±0.2°, 18.5±0.2°, 19.2±0.2°, 19.6±0.2°, 19.9±0.2°, 20.2±0.2°, 20.6±0.2°, 21.2±0.2°, 21.9±0.2°, 22.7±0.2°, 23.9±0.2°, 24.2±0.2°, 25.9±0.2°, 26.9±0.2°, 27.6±0.2°, 29.6±0.2°, 33.2±0.2°, 35.2±0.2°, and 40.8±0.2°, and their XRPD spectra are shown in Figure 31.

[0180] Comparative Example 14: Preparation of crystalline form R of 7,8-dihydroxyflavone Crystal form N was obtained by heating to 100°C using an in-situ thermochromic XRPD.

[0181] XRPD detection revealed the following X-ray powder diffraction values ​​expressed in 2θ angles: 9.7±0.2°, 10.8±0.2°, 11.1±0.2°, 11.7±0.2°, 12.4±0.2°, 12.6±0.2°, 12.8±0.2°, 13.4±0.2°, 13.9±0.2°, 14.1±0.2°, 14.3±0.2°, 14.5±0.2°, 15.6±0.2°, 16.1±0.2°, 16.4±0.2°, 17.5±0.2°, 19.2±0.2°, 19.7±0.2°, and 21.4±0.2°. Diffraction peaks were observed at 22.2±0.2°, 23.0±0.2°, 23.2±0.2°, 23.4±0.2°, 24.2±0.2°, 24.4±0.2°, 24.7±0.2°, 25.1±0.2°, 25.4±0.2°, 25.9±0.2°, 26.5±0.2°, 27.9±0.2°, 28.5±0.2°, 28.7±0.2°, 29.9±0.2°, 31.5±0.2°, 32.2±0.2°, and 44.3±0.2°, and their XRPD spectra are shown in Figure 32.

[0182] Comparative Example 15: Preparation of crystalline form S of 7,8-dihydroxyflavone 160 mg of 7,8-dihydroxyflavone was weighed out, and 11 mL of ethylene glycol dimethyl ether was added dropwise at room temperature to completely dissolve the sample. 1.3 mL of the solution was taken, and 8.0 mL of butyl formate was added dropwise. After stirring at room temperature for 15 min to 1 hour, the precipitated solid was centrifuged, and then the solid was vacuum-dried at room temperature.

[0183] XRPD detection revealed diffraction peaks in the X-ray powder diffraction, expressed as a 2θ angle, at 8.6±0.2°, 12.6±0.2°, 17.3±0.2°, 18.4±0.2°, 22.6±0.2°, 26.9±0.2°, 29.1±0.2°, 31.3±0.2°, 33.2±0.2°, 35.1±0.2°, 35.7±0.2°, 41.8±0.2°, and 44.8±0.2°. The XRPD spectra are shown in Figure 33.

[0184] Comparative Example 16: Preparation of crystalline form T of 7,8-dihydroxyflavone Crystal form G was obtained by heating to 140°C using an in-situ variable temperature XRPD.

[0185] XRPD detection revealed that the X-ray powder diffraction, expressed in 2θ angles, was 8.3±0.2°, 9.3±0.2°, 10.4±0.2°, 10.9±0.2°, 11.4±0.2°, 12.4±0.2°, 13.0±0.2°, 14.0±0.2°, 15.4±0.2°, 16.2±0.2°, 16.7±0.2°, 17.5±0.2°, 18.5±0.2°, 19.7±0.2°, 20.5±0.2°, 22.0±0.2°, 22.9±0.2°, 23.8±0.2°, 25.2±0.2°, 26.0±0.2°, 26.5±0.2°, Diffraction peaks were observed at 27.5±0.2°, 28.0±0.2°, 29.5±0.2°, 30.0±0.2°, 31.1±0.2°, 31.3±0.2°, 31.5±0.2°, 31.7±0.2°, 32.7±0.2°, 34.0±0.2°, 34.7±0.2°, 34.9±0.2°, 35.8±0.2°, 36.1±0.2°, 38.0±0.2°, 38.2±0.2°, 42.1±0.2°, 43.5±0.2°, 43.7±0.2°, and 44.1±0.2°, and their XRPD spectra are shown in Figure 34.

[0186] Tablet Preparation Example 1: Preparation of 7,8-dihydroxyflavone in tablet form 80g of the raw material (crystalline form of dihydroxyflavone prepared in this disclosure), 80g of lactose, 13g of microcrystalline cellulose, 12g of povidone, and 7g of sodium carboxymethyl starch were mixed and placed in a 1L granulation pan. The mixture was stirred for 6 minutes, 40g to 50g of pure water was added, and wet granulation was carried out, stirring for 8 to 10 minutes. After drying and granulation, 7g of sodium carboxymethyl starch and 1g of magnesium stearate were added, the materials were mixed, and then the tablets were compressed and demolded to obtain the target tablets.

[0187] Effect Example 1: Stability of different crystalline forms of 7,8-dihydroxyflavone Stability studies were conducted on different crystalline forms of 7,8-dihydroxyflavone under high temperature (60°C), high humidity (25°C / 92.5%RH), light irradiation (25°C / 4500Lux), and accelerated aging (40°C / 75%RH). The results of XRPD analysis performed on samples taken on days 7 and 15 are shown in Table 3, and the XPRD detection results are shown in Figures 35-37.

[0188] [Table 3]

[0189] Example 2: Dynamic solubility of different crystalline forms of 7,8-dihydroxyflavone in three biological media (FaSSIF, FeSSIF, and FaSSGF) and water. 20 mg of each crystalline form of 7,8-dihydroxyflavone was placed in 4.0 mL of biological medium or water, and the mixture was shaken at 37°C for 24 hours. Samples were taken at 0.5 h, 2 h, and 24 h, and the sampled solutions were filtered through a 0.22 μm aqueous filtration membrane. The signal peak area of ​​the solution was measured by HPLC, and finally, the concentration of the compound in the solution was calculated based on the peak area, the HPLC calibration curve of the raw materials, and the dilution factor. The supernatant at 24 h was also taken and its pH was tested, and the remaining solid was subjected to XRPD testing. The composition of the biological medium is shown in Table 4. The experimental results are shown in Table 5, and the XPRD detection results are shown in Figures 38-40.

[0190] [Table 4]

[0191] [Table 5]

[0192] Example of effect 3: Stability study of tablets containing crystalline form P of 7,8-dihydroxyflavone Stability studies were conducted on tablets of 7,8-dihydroxyflavone in crystalline form P under high temperature (60°C), high humidity (25°C / 92.5%RH), light irradiation (25°C / 4500 Lux), and accelerated aging (40°C / 75%RH). Samples were taken at 3 months and 6 months, respectively, and XRPD analysis was performed. The results are shown in Table 6.

[0193] [Table 6]

[0194] Example of effect 4: Stability study of tablets containing crystalline form C of 7,8-dihydroxyflavone Stability studies were conducted on tablets of 7,8-dihydroxyflavone in crystalline form C under high temperature (60°C), high humidity (25°C / 92.5%RH), light irradiation (25°C / 4500 Lux), and accelerated aging (40°C / 75%RH). Samples were taken at 3 months and 6 months, and XRPD analysis was performed. The results are shown in Table 7.

[0195] [Table 7]

[0196] Example of effect 5: Dissolution rate study of tablets containing crystalline form P of 7,8-dihydroxyflavone Dissolution studies were conducted on tablets of 7,8-dihydroxyflavone in crystalline form P under different pH conditions, and the results are shown in Table 8.

[0197] [Table 8]

[0198] Example 6: Dissolution rate study of tablets containing crystalline form C of 7,8-dihydroxyflavone Dissolution studies were conducted on tablets of crystalline form C of 7,8-dihydroxyflavone under different pH conditions, and the results are shown in Table 9.

[0199] [Table 9]

[0200] Example of effect 7: Dissolution rate study of tablets containing crystalline form A of 7,8-dihydroxyflavone Dissolution studies were conducted on tablets of crystalline form A of 7,8-dihydroxyflavone under different pH conditions, and the results are shown in Table 10.

[0201] [Table 10]

[0202] In summary, this disclosure provides crystalline polymorphs of 7,8-dihydroxyflavone and methods for preparing them. The crystalline polymorphs of 7,8-dihydroxyflavone according to this disclosure have a greater range of selectable pharmacoeconomic values.

Claims

1. The X-ray powder diffraction pattern has characteristic peaks at approximately 8.3 ± 0.2°, 25.0 ± 0.2°, and 25.9 ± 0.2° at a 2θ angle. Crystalline form P of 7,8-dihydroxyflavone.

2. The X-ray powder diffraction pattern further includes characteristic peaks at approximately 27.4 ± 0.2° and / or 31.4 ± 0.2°. Crystalline form P of 7,8-dihydroxyflavone as described in claim 1.

3. The X-ray powder diffraction pattern has characteristic peaks at angles of approximately 11.1±0.2°, 15.6±0.2°, 27.0±0.2°, and 28.3±0.2° at 2θ angles. Crystalline form C of 7,8-dihydroxyflavone.

4. The X-ray powder diffraction pattern further includes characteristic peaks at approximately 5.6±0.2°, 9.5±0.2°, 15.5±0.2°, 21.0±0.2°, 24.3±0.2° and / or 25.5±0.2°. Crystalline form C of 7,8-dihydroxyflavone as described in claim 3.

5. It is a hydrate, for example, a monohydrate. The crystalline form C of 7,8-dihydroxyflavone according to any one of claims 3 to 4.

6. The X-ray powder diffraction pattern has characteristic peaks at angles of approximately 9.3±0.2°, 23.8±0.2°, 24.6±0.2°, and 26.6±0.2° at 2θ angles. Crystalline form N of 7,8-dihydroxyflavone.

7. The X-ray powder diffraction pattern further includes characteristic peaks at approximately 12.3 ± 0.2° and / or 21.1 ± 0.2°. Crystalline form N of 7,8-dihydroxyflavone according to claim 6.

8. It is a hydrate, for example, a dihydrate. Crystalline form N of 7,8-dihydroxyflavone according to any one of claims 6 to 7.

9. (1) a crystalline form P of 7,8-dihydroxyflavone according to any one of claims 1 to 2, a crystalline form C of 7,8-dihydroxyflavone according to any one of claims 3 to 5, and / or a crystalline form N of 7,8-dihydroxyflavone according to any one of claims 6 to 8, and (2) at least one pharmaceutically acceptable excipient, Pharmaceutical composition.

10. A method for treating diseases associated with abnormalities in the TrkB signaling pathway, such as central nervous system injury, peripheral nerve injury, neurodegenerative lesions, mental disorders, hereditary neurological disorders, ophthalmic diseases, metabolic diseases, pain, cardiovascular diseases, tumors, or other related diseases, (1) administering to a subject crystalline form P of 7,8-dihydroxyflavone according to any one of claims 1 to 2, crystalline form C of 7,8-dihydroxyflavone according to any one of claims 3 to 5, and / or crystalline form N of 7,8-dihydroxyflavone according to any one of claims 6 to 8, and (2) at least one pharmaceutically acceptable excipient. method.

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