MGLUR5 Modulating Compounds, Compositions, and Methods of Use

Abstract

The present disclosure provides compositions of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridin-3-yl)oxazolidin-2-one (Compound 1). Crystal and solvate forms of the compound are also provided, as well as formulations comprising the compound. Methods of using the compounds and administering formulations to a subject in need thereof to treat or prevent CNS disorders, such as Alzheimer's disease, are provided.

Description

[Technical field]

[0001] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with Government support under grant U01 AG058608 awarded by the National Institutes of Health. The Government has certain rights in this invention. [Background technology]

[0002] Glutamate is the main excitatory neurotransmitter in the brain and is involved in various psychiatric and medical conditions. Glutamate regulates the function of the central nervous system through the operation of ionotropic and metabotropic receptors. Three groups of metabotropic (mGlu) receptors regulate neuronal activity through G protein-coupled signaling. In particular, the mGluR5 receptor, a group I receptor, plays an important role in mental health and various central nervous system (CNS) disorders. Several mGluR5 antagonists have been designed, including 3-[[2-methyl-1,3-thiazol-4-yl]ethynyl]pyridine (MTEP), acamprosate, memantine, AFQ056, and fenobam. Allosteric modulators of mGluR5 can be classified into positive allosteric modulators (PAMs), negative allosteric modulators (NAMs), and silent allosteric modulators (SAMs).

[0003] There remains a need for compounds and compositions that modulate mGluR5 to improve mental health and ameliorate CNS disorders. Summary of the Invention

[0004] In one aspect of this disclosure, a pharmaceutical composition is provided that includes (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridin-3-yl)oxazolidin-2-one (Compound 1), of the following formula: In some embodiments, the pharmaceutical composition includes Compound 1, lactose monohydrate, croscarmellose sodium, and / or magnesium stearate. [ka]

[0005] In another aspect of this disclosure, a method for treating a CNS disorder is provided by administering a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof, to a subject in need thereof. In some embodiments, the CNS disorder is Alzheimer's disease. In some embodiments, a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof, is used to prevent convulsions, restore synaptic loss, or reduce tau accumulation in a subject in need thereof.

[0006] The present disclosure further provides crystalline forms, such as anhydrous crystalline forms, of Compound 1. The present disclosure further provides solvates, such as the isopropyl solvate, of Compound 1.

[0007] In one aspect, the invention features an anhydrous (AH) crystalline form of Compound 1 (Form A) having an XRPD pattern including peaks at 2θ angles of 9.02, 11.65, and 11.86 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern further includes peaks at 2θ angles of 12.15, 14.99, 28.99, and 44.03 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern further includes peaks at three 2θ angles of 21.09, 21.49, and 21.88 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern comprises peaks at 2θ angles of 9.02, 11.65, 11.86, 12.15, 14.99, 21.09, 21.49, 21.88, 28.99, and 44.03 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern is substantially as shown in FIG.

[0008] In some embodiments, Form AH has a melting endotherm peak in a differential scanning calorimetry (DSC) thermogram at about 134° C.-138° C. In some embodiments, Form AH has a DSC thermogram substantially the same as the DSC graph shown in FIG. 4. In some embodiments, Form AH has a thermogravimetric analysis (TGA) weight loss of about 0.02% (w / w) between ambient temperature and about 250° C. In some embodiments, Form AH has a TGA substantially the same as the TGA graph shown in FIG. 4. In some embodiments, Form AH is substantially purified.

[0009] In one aspect, the invention features a method of preparing an anhydrous (AH) crystalline form (Form A) of Compound 1, the method includes precipitating the anhydrous crystalline form from a solution including Compound 1 and a solvent selected from the group consisting of ethyl acetate (EtOAc), heptane, and mixtures thereof. In some embodiments, the solvent is a mixture of EtOAc and heptane. In some embodiments, the ratio of EtOAc to heptane is 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, or 10:90. In some embodiments, the method further includes cooling the solution. In some embodiments, the method further includes isolating the AH crystalline form by filtration.

[0010] In one aspect, the invention features a solvate form of Compound 1 (Form B) having an XRPD pattern including at least three of the following peaks at 2θ angles, 16.07, 16.27, 16.58, and 16.77 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern further includes three or more of the following peaks at 2θ angles, 13.18, 13.4, 13.54, 13.54, and 13.70 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern further includes peaks at 2θ angles, 37.31, and 39.92 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern comprises peaks at 2θ angles of 13.18, 13.4, 13.54, 13.54, 13.70, 16.07, 16.27, 16.58, 16.77, 37.31, and 39.92 degrees 2θ, each ±0.2 degrees 2θ, when measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, solvate Form B has an XRPD pattern substantially as shown in FIG.

[0011] In some embodiments, solvate form B has a melting endotherm peak at about 134° C.-138° C. in a differential scanning calorimetry (DSC) thermogram. In some embodiments, solvate form B has a DSC thermogram substantially the same as the DSC graph shown in FIG. 6. In some embodiments, solvate form B has a thermogravimetric analysis (TGA) weight loss of about 6.7% (w / w) between ambient and about 200° C. In some embodiments, solvate form B has a TGA substantially the same as the TGA graph shown in FIG. 6. In some embodiments, solvate form B is an isopropyl alcohol solvate. In some embodiments, solvate form B is substantially purified.

[0012] In one aspect, the invention features a method of preparing solvate form B of Compound 1, the method includes evaporating a mixture of Compound 1 and isopropyl alcohol under an inert atmosphere in a dry box. In some embodiments, the evaporation occurs over a period of 1-2 weeks. In some embodiments, the method further includes drying solvate form B under vacuum.

[0013] In one aspect, the invention features a solvate form of Compound 1 (Form C) having an XRPD pattern that includes only two peaks between 13.32 and 13.82 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern includes only three peaks between 32.22 and 32.96 degrees 2θ, each ±0.2 degrees 2θ, or includes peaks at 3.4, 8.33, 10.94, 16.32, and 16.66 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, the XRPD pattern comprises peaks at 2θ angles of 3.4, 8.33, 10.94, 13.32, 13.82, 16.32, 16.66, 32.22, 32.59, 32.96 degrees 2θ, each ±0.2 degrees 2θ, as measured by X-ray diffraction measurement with Cu Kα X-ray radiation. In some embodiments, solvate form C has an XRPD pattern substantially as shown in Figure 7. In some embodiments, solvate form C has an XRPD pattern substantially as shown in Figure 9.

[0014] In some embodiments, solvate form C has a melting endotherm peak at about 129-133° C. in a differential scanning calorimetry (DSC) thermogram. In some embodiments, solvate form C has a DSC thermogram substantially the DSC graph shown in FIG. 8. In some embodiments, solvate form C has a thermogravimetric analysis (TGA) weight loss of about 3.4% (w / w) between ambient and about 200° C. In some embodiments, solvate form C has a TGA substantially the TGA graph shown in FIG. 8. In some embodiments, form C is an isopropyl alcohol solvate. In some embodiments, form C is substantially purified.

[0015] In one aspect, the invention features a method of preparing solvate form C of Compound 1 using extended slurry. In some embodiments, the method further comprises precipitating the solvate form from a solution comprising Compound 1 and isopropyl alcohol. In some embodiments, the method further comprises stirring the solution at 20° C. for 2 weeks. In some embodiments, the method further comprises isolating solvate form C by filtration.

[0016] In some embodiments, solvate form C has endothermic peaks at about 89.4° C. and about 134.2° C. in a differential scanning calorimetry (DSC) thermogram. In some embodiments, solvate form C has a DSC thermogram substantially the same as the DSC graph shown in FIG. 10. In some embodiments, solvate form C has a thermogravimetric analysis (TGA) weight loss of about 3.2% (w / w) between ambient temperature and about 200° C. In some embodiments, solvate form C has a TGA substantially the same as the TGA graph shown in FIG.

[0017] In one aspect, the invention features a method of preparing solvate Form C of Compound 1 using vapor diffusion. In some embodiments, the method further comprises: (i) dissolving compound 1 in isopropyl alcohol to form a saturated solution; (ii) filtering the mixture of step (i) into a vial that is placed within an outer vial containing the anti-solvent; (iii) agitating the mixture in the inner vial of step (ii) to allow the solids to settle; and (iv) filtering the solid from step (iii) and drying the solid under vacuum at room temperature. Includes.

[0018] In some embodiments, the anti-solvent is pentane.

[0019] In some embodiments, the pharmaceutical composition of the present invention comprises a therapeutically effective amount of the crystalline or solvated form of the present invention. In some embodiments, the pharmaceutical composition comprises between about 5% (w / w) and about 30% (w / w) of the crystalline or solvated form of the present invention. In some embodiments, the pharmaceutical composition comprises about 5% (w / w), 10% (w / w), 24% (w / w), or about 25% (w / w) of the crystalline or solvated form of the present invention. In some embodiments, the pharmaceutical composition of the present invention is a nanosuspension or a spray-dried nanosuspension. In some embodiments, the pharmaceutical composition of the present invention further comprises a pharma- ceutically acceptable plasticizer, binder, filler, carrier, excipient, lubricant, disintegrant, and / or surfactant. In some embodiments, the pharmaceutical composition further comprises at least one of hypromellose, sodium lauryl sulfate, and lactose monohydrate. In some embodiments, the pharmaceutical composition further comprises at least one of croscarmellose and magnesium stearate. In some embodiments, the pharmaceutical composition is in capsule form. In some embodiments, the capsule is selected from a hard hydroxypropyl methylcellulose capsule, a hard gelatin capsule, or a soft gelatin capsule. In some embodiments, the pharmaceutical composition of the present invention comprises the ingredients shown in the table below. JPEG2025509988000002.jpg5068, provided that the amounts of binder, surfactant, carrier, disintegrant, and lubricant are not all 0%. In some embodiments, the binder is hypromellose, the surfactant is sodium lauryl sulfate, the carrier and binder is lactose monohydrate, the disintegrant is croscarmellose, and the lubricant is magnesium stearate.

[0020] In some embodiments, the pharmaceutical composition comprises a nanosuspension of Compound 1 (e.g., Form A, Form B, or Form C) having a x90 of <400 μm, e.g., a x90 of about 300, a x50 of about 135, and / or a x10 of 70, as measured by PSD. In some embodiments, the pharmaceutical composition comprises a carrier (e.g., lactose) onto which the nanosuspension of Compound 1 (e.g., Form A, Form B, or Form C) has been sprayed, e.g., in combination with a bulking agent (e.g., lactose), to form granules.

[0021] In one aspect, the invention features a method for treating Alzheimer's disease, preventing seizures, reversing synaptic loss, or reducing tau accumulation in a subject in need thereof, comprising treating the subject with a therapeutically effective amount of Form A, Form B, or Form D of Compound 1. In some embodiments, about 10 mg, 40 mg, 70 mg, 100 mg, 150 mg, or 200 mg of Compound 1 is administered. In some embodiments, Compound 1 is administered orally. [Brief description of the drawings]

[0022] The foregoing and other objects, features, and advantages will become apparent from the following description of specific embodiments of the present disclosure and are illustrated in the accompanying drawings, in which like reference characters refer to like parts in the different drawings. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of various embodiments of the present disclosure.

[0023] [Figure 1] FIG. 1 shows the dose response of compound 1 displacement of [18F]FPEB. [Diagram 2] FIG. 2 shows that Compound 1 treatment prevents PAM-induced convulsions. [Diagram 3] FIG. 3 shows the XRPD data of Compound 1, Form AH. [Figure 4] Figure 4 shows the TGA / DSC data for AH Form A of Compound 1. The TGA showed a decomposition event above 250°C. The DSC trace showed a single endothermic event with an onset at 135.1°C and a peak apex at 136.5°C due to melting of the crystalline material. [Diagram 5] FIG. 5 shows the XRPD data for Form B of Compound 1. [Figure 6] 6 is the TGA / DSC data for Form B of Compound 1. The TGA showed a desolvation event occurring up to about 200° C. The DSC trace showed an endothermic event with a first onset at about 83.4° C. and peak apex at 90.3° C. due to desolvation, and a second onset at 133.8° C. and peak apex at 136.0° C. due to melting of the crystalline material. [Figure 7] FIG. 7 shows XRPD data for Form C of Compound 1 prepared from an extended slurry using isopropyl alcohol. [Figure 8] 8 shows the TGA / DSC data for Form C of Compound 1 prepared from an extended slurry using isopropyl alcohol. The TGA showed a desolvation event that occurred up to about 200° C. The DSC trace showed a first endothermic event due to desolvation with an onset at 86.1° C. and apex at a peak at 89.1° C., and a second endothermic event due to melting of the crystalline material with an onset at 129.1° C. and apex at a peak at 131.7° C. [Figure 9] FIG. 9 shows the XRPD data of Form C of Compound 1 prepared by vapor diffusion using isopropyl alcohol. [Figure 10]10 shows the TGA / DSC data for Form C of Compound 1 prepared by vapor diffusion using isopropyl alcohol. The TGA showed a desolvation event that occurred at approximately 200° C. The DSC trace showed a first endothermic event with an onset at 78.7.1° C. and a peak apex at 89.4° C., and a second endothermic event with an onset at 132.7° C. and a peak apex at 134.2° C. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] The present disclosure provides compounds and compositions for modulating mGluR5 receptors, as well as methods of using these compounds and compositions. mGluR5 receptors are widely expressed throughout the CNS, with predominantly postsynaptic localization, but also presynaptic expression. It is a Gαq-coupled receptor that activates phospholipase C, elevates intracellular calcium levels, and activates downstream signaling molecules. Many studies have demonstrated the role of this receptor in regulating N-methyl-D-aspartate (NMDA) receptor activity and synaptic plasticity, suggesting that this receptor plays an important role in glutamate signaling.

[0025] The compounds of the present disclosure bind to mGluR5 and are allosteric modulators of mGluR5.In some embodiments, the compounds are mGluR5 silent allosteric modulators.Thus, the compounds and compositions described herein can be administered to regulate mGluR5 and treat an individual subject with mGluR5-related diseases, disorders and / or conditions, including but not limited to CNS disorders.

[0026] I. Compounds of the Disclosure The compound of the present disclosure is (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridin-3-yl)oxazolidin-2-one (Compound 1), of the formula below, or a pharma- ceutically acceptable salt thereof, which is described in U.S. Pat. No. 8,691,821 and is incorporated herein by reference. [ka]

[0027] The compounds of the present disclosure may be prepared using any convenient method well known to those skilled in the art, for example, the methods disclosed in US Pat. No. 8,691,821.

[0028] Solid forms of compound 1 Solids exist in either amorphous or crystalline form. Polymorphism refers to the different crystalline forms of a chemical substance. These solid forms and crystalline forms are characterized by different structures and physical properties such as XRPD spectra, IR spectra, melting point, etc. Certain solid forms or polymorphs may be superior to other forms and may be more suitable for the manufacture and use of a drug substance.

[0029] An anhydrous (AH) form (Form A), two solvate forms (solvates S1 and S2, interchangeably referred to as Forms B and C, respectively), and multiple amorphous forms of Compound 1 have been discovered. The amorphous forms gel after evaporation in a variety of solvent systems. Anhydrous Form A is the most stable form, as determined, for example, by DSC and TGA analysis.

[0030] Form AH can be identified by single crystal X-ray powder diffraction (XRPD) using Cu Kα X-ray radiation. The major peaks of Form A were identified and their relative intensities are shown in Table 1. As one of ordinary skill in the art would understand, the relative intensities of the peaks in Table 1 may vary depending on various factors, such as the purity of the material analyzed, the effect of crystal orientation in the X-ray beam, and the degree of crystallization of the sample. Peak positions may shift due to variations in sample height, but the peak positions will remain substantially as defined in Table 1. One of ordinary skill in the art would also understand that measurements using different wavelengths will result in different shifts according to the Bragg equation (nλ=2d sin θ). Such additional XRPD patterns generated using alternative wavelengths are alternative representations of the XRPD pattern of a crystalline material.

[0031] Table 1. XRPD peak list for Form A [Table 1] JPEG2025509988000005.jpg11

[0032] Form A may be characterized by any of the peaks in Table 1. For example, Form A may be characterized by any of the following peaks, among others: 6.75, 6.88, 9.02, 11.65, 11.86, 12.15, 13.55, 14.99, 15.59, 17.55, 17.97, 18.45, 18.93, 19.46, 20.34, 21.09, 21.49, 21.88, 22.29, 22.75, 23.59, 24.24, 24.54, 25.08, 25.63, 26.14, 26.44, 26.81, 27.12, 27.53, 27.97, 28.41, 28. 0.99, 29.45, 29.83, 30.28, 30.62, 30.87, 31.15, 31.65, 32.25, 32.82, 33.13, 33.29, 34.23, 34.93, 35.76, 36.66, 37.86, 38.64, 39.02, 40.43, 40.99, 41.68, 42.32, or 44.03 degrees 2θ, each ±0.2 degrees 2θ. Form A may be characterized, among others, by any of the following d-spacings: 13.08, 12.84, 9.80, 7.59, 7.46, 7.28, 6.53, 5.90, 5.68, 5.05, 4.93, 4.81, 4.69, 4.56, 4.36, 4.21, 4.13, 4.06, 3.99, 3.91, 3.77, 3.67, 3.63, 3.55, 3.47, 3.41, 3.37, 3.3 2, 3.29, 3.24, 3.19, 3.14, 3.08, 3.03, 2.99, 2.95, 2.92, 2.89, 2.87, 2.82, 2.77, 2.73, 2.70, 2.69, 2.62, 2.57, 2.51, 2.45, 2.37, 2.33, 2.31, 2.23, 2.20, 2.17, 2.13, 2.05 Å, each ±0.2 Å. Form A may also be characterized by an XRPD pattern substantially as shown in FIG.

[0033] In some embodiments, the anhydrous crystalline form of Compound 1 is Form A and is 6.75, 6.88, 9.02, 11.65, 11.86, 12.15, 13.55, 14.99, 15.59, 17.55, 17.97, 18.45, 18.93, 19.46, 20.34, 21.09, 21.49, 21.88, 22.29, 22.75, 23.59, 24.24, 24.54, 25.08, 25.63, 26.14, 26.44, 26.81, 27.12, 27.53, 35.76, 36.66, 37.86, 38.64, 39.02, 40.43, 40.99, 41.68, 42.32, or 44.03 degrees 2θ, each ±0.2 degrees 2θ.

[0034] In some embodiments, the anhydrous crystalline form of Compound 1 is Form A, and ... 0.53, 27.97, 28.41, 28.99, 29.45, 29.83, 30.28, 30.62, 30.87, 31.15, 31.65, 32.25, 32.82, 33.13, 33.29, 34.23, 34.93, 35.76, 36.66, 37.86, 38.64, 39.02, 40.43, 40.99, 41.68, 42.32, and 44.03 degrees 2θ, each ±0.2 degrees 2θ.

[0035] In some embodiments, Form A, as analyzed by differential scanning calorimetry (DSC) thermogram, exhibits a melting endotherm peak at about 134° C. to 138° C., as shown in FIG.

[0036] In some embodiments, Form A analyzed by thermogravimetric analysis (TGA) exhibits dehydration from ambient temperature to about 250° C. with a weight loss of about 0.02% (w / w), as shown in FIG.

[0037] In some embodiments, a method is provided for preparing an anhydrous (AH) crystalline form of Compound 1, comprising precipitating the anhydrous crystalline form from a solution comprising Compound 1 and a solvent selected from the group of ethyl acetate (EtOAc), heptane, and mixtures thereof. In some embodiments, the solvent can be a mixture of EtOAc and heptane. The ratio of EtOAc to heptane is 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, or 10:90. The method can further include cooling the solution and / or isolating the AH crystalline form by filtration.

[0038] Form B can be identified by single crystal X-ray powder diffraction (XRPD) with Cu Kα X-ray radiation. The major peaks of Form B were identified and their relative intensities are shown in Table 2. As one of ordinary skill in the art would understand, the relative intensities of the peaks in Table 2 may vary depending on various factors, such as the purity of the material analyzed, the effect of crystal orientation in the X-ray beam, and the degree of crystallization of the sample. Peak positions may shift due to variations in sample height, but the peak positions will remain substantially as defined in Table 2. One of ordinary skill in the art would also understand that measurements using different wavelengths will result in different shifts according to the Bragg equation (nλ=2d sin θ). Such additional XRPD patterns generated using alternative wavelengths are alternative representations of the XRPD pattern of a crystalline material.

[0039] Table 2. XRPD peak list for Form B [Table 2] JPEG2025509988000007.jpg11682

[0040] Form B may be characterized by any of the peaks in Table 2. For example, Form B may be characterized by any of the following peaks, among others: 3.36, 6.75, 6.85, 8.19, 10.14, 10.84, 13.18, 13.4, 13.54, 13.7, 15.65, 16.07, 16.27, 16.58, 16.77, 17.15, 17.74, 18.63, 18.73, 19.16, 19.44, 19.63, 19.95, 20.21, 20.36, 20.6, 21.56, 21.78, 22.42, 22.88, 23.06, 23.22, 23.43, 23.75, 23.99, 24.53, 24.88, 25.23, 25.43, 26.03, 26.63, 26.89, 27.27, 27.7, 28.22, 29.44, 29.95, 30.24, 30.51, 30.71, 31.14, 31.72, 32.01, 32.27, 32.47, 32.85, 33.48, 33.62, 34.15, 34.63, 35.23, 35.91, 36.16, 36.71, 37.31, 37.79, 38.2, 38.83, 39.92, 40.26, 40.58, 41.37, 42.56, 43.38, or 44.67 degrees 2θ, each ±0.2 degrees 2θ. Form B may be characterized, among others, by any of the following d-spacings: 26.31, 13.09, 12.9, 10.79, 8.72, 8.15, 6.71, 6.6, 6.54, 6.46, 5.66, 5.51, 5.45, 5.34, 5.28, 5.17, 4.99, 4.76, 4.73, 4.63, 4.56, 4.52, 4.45, 4.39, 4.36, 4.31, 4.12, 4.08, 3.96, 3.88, 3.85, 3.83, 3.79, 3.74, 3.71, 3.63, 3.58, 3.53 , 3.5, 3.42, 3.34, 3.31, 3.27, 3.22, 3.16, 3.03, 2.98, 2.95, 2.93, 2.91, 2.87, 2.82, 2.79, 2.77, 2.75, 2.72, 2.67, 2.66, 2.62, 2.59, 2.55, 2.5, 2.48, 2.45, 2.41, 2.38, 2.35, 2.32, 2.26, 2.24, 2.22, 2.18, 2.12, 2.08, or 2.03, each ±0.2 Å. Form B may also be characterized by an XRPD pattern substantially as shown in FIG.

[0041] In some embodiments, the solvate form of Compound 1 is Form B, and includes the following: 3.36, 6.75, 6.85, 8.19, 10.14, 10.84, 13.18, 13.4, 13.54, 13.7, 15.65, 16.07, 16.27, 16.58, 16.77, 17.15, 17.74, 18. 63, 18.73, 19.16, 19.44, 19.63, 19.95, 20.21, 20.36, 20.6, 21.56, 21.78, 22.42, 22.88, 23.06, 23.22, 23.43, 23.75, 23.99, 24.53, 24.88, 25.23, 25.43, 26.0 3, 26.63, 26.89, 27.27, 27.7, 28.22, 29.44, 29.95, 30.24, 30.51, 30.71, 31.14, 31.72, 32.01, 32.27, 32.47, 32.85, 33.48, 33.62, 34.15, 34.63, 35.23, 35.91, 36.16, 36.71, 37.31, 37.79, 38.2, 38.83, 39.92, 40.26, 40.58, 41.37, 42.56, 43.38, or 44.67 degrees 2θ, each ±0.2 degrees 2θ.

[0042] In some embodiments, the solvate form of Compound 1 is Form B and is: 3.36, 6.75, 6.85, 8.19, 10.14, 10.84, 13.18, 13.4, 13.54, 13.7, 15.65, 16.07, 16.27, 16.58, 16.77, 17.15, 17.74, 18.63, 18.73, 19.16, 19.44, 19.63, 19.95, 20.21, 20.36, 20.6, 21.56, 21.78, 22.42, 22.88, 23.06, 23.22, 23.43, 23.75, 23.99, 24.53, 24.88, 25.23, 25.43, It has an X-ray powder diffraction (XRPD) pattern which includes all peaks at 26.03, 26.63, 26.89, 27.27, 27.7, 28.22, 29.44, 29.95, 30.24, 30.51, 30.71, 31.14, 31.72, 32.01, 32.27, 32.47, 32.85, 33.48, 33.62, 34.15, 34.63, 35.23, 35.91, 36.16, 36.71, 37.31, 37.79, 38.2, 38.83, 39.92, 40.26, 40.58, 41.37, 42.56, 43.38, and 44.67 degrees 2θ, each ±0.2 degrees 2θ.

[0043] In some embodiments, Form B, as analyzed by differential scanning calorimetry (DSC) thermogram, exhibits a melting endotherm peak at about 134° C.-138° C., as shown in FIG.

[0044] In some embodiments, Form B analyzed by thermogravimetric analysis (TGA) exhibits desolvation with a weight loss of about 6.7% (w / w) from ambient temperature to about 200° C., as shown in FIG.

[0045] In some implementations, a method for preparing Form B of Compound 1 is provided that includes evaporating a mixture of Compound 1 and isopropyl alcohol under an inert atmosphere, such as nitrogen gas, in a dry box. In some embodiments, the evaporation may occur over a period of 1 to 2 weeks. In some embodiments, the method may further include drying crystalline form B under vacuum.

[0046] Form C prepared from an extended slurry in isopropyl alcohol can be characterized by single crystal XRPD with Cu Kα X-ray irradiation. The major peaks of Form C were identified and their relative intensities are shown in Table 3. As one of ordinary skill in the art would understand, the relative intensities of the peaks in Table 3 may vary depending on various factors, such as the purity of the material analyzed, the effect of crystal orientation in the X-ray beam, and the degree of crystallization of the sample. Peak positions may shift due to variations in sample height, but the peak positions will remain substantially as defined in Table 3. One of ordinary skill in the art would also understand that measurements using different wavelengths will result in different shifts according to the Bragg equation (nλ=2d sin θ). Such additional XRPD patterns generated using alternative wavelengths are alternative representations of the XRPD pattern of a crystalline material.

[0047] Table 3. XRPD peak list for Form C prepared from extended slurry with isopropyl alcohol. [Table 3]

[0048] Form C prepared from extended slurry with isopropyl alcohol may be characterized by any of the peaks in Table 3. For example, Form C may be characterized by any of the following peaks, among others: 3.40, 6.83, 8.33, 10.94, 13.32, 13.82, 16.32, 16.66, 16.95, 17.24, 18.76, 19.27, 19.6, 20.13, 20.31, 20.81, 21.86, 22.72, 23.15, 23.38, 23.57, 23.86, 24.30, 24.89. , 25.61, 26.24, 26.92, 27.28, 28.53, 29.51, 30.63, 31.64, 32.22, 32.59, 32.96, 33.72, 34.34, 34.8, 35.36, 36.25, 36.91, 38.24, 42.75, 43.52, or 44.99 degrees 2θ, each ±0.2 degrees 2θ. Form C prepared from an extended slurry in isopropyl alcohol may be characterized, among other things, by any of the following d-spacings: 25.93, 12.92, 10.61, 8.08, 6.64, 6.4, 5.43, 5.32, 5.23, 5.14, 4.73, 4.6, 4.53, 4.41, 4.37, 4.26, 4.06, 3.91, 3.84, 3.8, 3.77, 3.73, 3.66, 3.57, 3.48, 3.39, 3.31, 3.27, 3.13, 3.02, 2.92, 2.83, 2.78, 2.75, 2.72, 2.66, 2.61, 2.58, 2.54, 2.48, 2.43, 2.35, 2.11, 2.08, or 2.01, each ±0.2 Å. Form C prepared from an extended slurry using isopropyl alcohol may also be characterized by an XRPD pattern substantially as shown in FIG.

[0049] In some embodiments, the solvate form of Compound 1 is Form C prepared from extended slurry with isopropyl alcohol and has the following concentrations: 3.40, 6.83, 8.33, 10.94, 13.32, 13.82, 16.32, 16.66, 16.95, 17.24, 18.76, 19.27, 19.6, 20.13, 20.31, 20.81, 21.86, 22.72, 23.15, 23.38, 23.57, 23.8 6, 24.30, 24.89, 25.61, 26.24, 26.92, 27.28, 28.53, 29.51, 30.63, 31.64, 32.22, 32.59, 32.96, 33.72, 34.34, 34.8, 35.36, 36.25, 36.91, 38.24, 42.75, 43.52, or 44.99 degrees 2θ, each ±0.2 degrees 2θ.

[0050] In some embodiments, the solvate form of Compound 1 is Form C prepared from extended slurry with isopropyl alcohol and has the following concentrations: 3.40, 6.83, 8.33, 10.94, 13.32, 13.82, 16.32, 16.66, 16.95, 17.24, 18.76, 19.27, 19.6, 20.13, 20.31, 20.81, 21.86, 22.72, 23.15, 23.38, 23.57, 23.06, 23.04, 23.08, 23.06, 23.08, 23.09, 24.06, 24.08, 24.09, 25.09, 26.06, 27.09, 28.09, 29.09, 30.09, 31.09, 32.09, 33.09, 34.09, 35.09, 36.09, 37.09, 38.09, 39.09, 40.09, 41.09, 42.09, 43.09, 44.09, 45.09, 46.09, 47.09, 48.09, 49.09, 50.09, 51.09, 52.09, 53.09, 54.09, 55.09, 56.09, 57.09, 58.09, 59.09, 60.09, 61.09, 62.09, 63.09, 30.63, 31.64, 32.22, 32.59, 32.96, 33.72, 34.34, 34.8, 35.36, 36.25, 36.91, 38.24, 42.75, 43.52, and 44.99 degrees 2θ, each ±0.2 degrees 2θ.

[0051] In some embodiments, Form C prepared from extended slurry with isopropyl alcohol, as analyzed by differential scanning calorimetry (DSC) thermogram, exhibits a melting endotherm peak at about 129-133° C., as shown in FIG. 8.

[0052] In some embodiments, Form C prepared from extended slurry with isopropyl alcohol, analyzed by thermogravimetric analysis (TGA), exhibits dehydration with a weight loss of about 3.4% (w / w) from ambient temperature to about 200° C., as shown in FIG.

[0053] In some implementations, a process is provided for preparing Form C of Compound 1, comprising precipitating the solvate form from a solution comprising Compound 1 and isopropyl alcohol. In some embodiments, the solution may be stirred at 10-30° C., e.g., 20° C., for 2 weeks. The process may further comprise isolating the solvate by filtration.

[0054] Form C prepared by vapor diffusion using isopropyl alcohol can be characterized by single crystal XRPD with Cu Kα X-ray irradiation. The major peaks of Form C were identified and their relative intensities are shown in Table 4. As one of ordinary skill in the art would understand, the relative intensities of the peaks in Table 4 may vary depending on various factors, such as the purity of the material analyzed, the effect of crystal orientation in the X-ray beam, and the degree of crystallization of the sample. Peak positions may shift due to variations in sample height, but the peak positions will remain substantially as defined in Table 4. One of ordinary skill in the art would also understand that measurements using different wavelengths will result in different shifts according to the Bragg equation (nλ=2d sin θ). Such additional XRPD patterns generated using alternative wavelengths are alternative representations of the XRPD pattern of a crystalline material.

[0055] Table 4. XRPD peak list for Form C prepared by vapor diffusion using isopropyl alcohol [Table 4]

[0056] Form C prepared by vapor diffusion using isopropyl alcohol may be characterized by any of the peaks in Table 4. For example, Form C may be characterized by any of the following peaks, among others: 3.38, 6.85, 8.35, 10.95, 13.31, 13.84, 16.31, 16.65, 16.99, 17.23, 18.76, 19.33, 19.6, 20.13, 20.30, 20.80, 21.05, 21.86, 22.68, 23.13, 23.6, 23.88, 24.3 , 24.9, 25.64, 26.24, 26.88, 27.26, 28, 28.53, 29.53, 30.66, 31.64, 32.59, 32.95, 33.74, 34.78, 36.21, 36.94, 38.24, 40.17, 40.80, or 43.55 degrees 2θ, each ±0.2 degrees 2θ. Form C prepared by vapor diffusion using isopropyl alcohol may be characterized, among other things, by any of the following d-spacings: 26.12, 12.9, 10.58, 8.07, 6.65, 6.39, 5.43, 5.32, 5.21, 5.14, 4.73, 4.59, 4.53, 4.41, 4.37, 4.27, 4.22, 4.06, 3.92, 3.84, 3.77, 3.72, 3.66, 3.57, 3.47, 3.39, 3.31, 3.27, 3.18, 3.13, 3.02, 2.91, 2.83, 2.75, 2.72, 2.65, 2.58, 2.48, 2.43, 2.35, 2.24, 2.21, or 2.08, each ±0.2 Å. Form C prepared by vapor diffusion using isopropyl alcohol may also be characterized by an XRPD pattern substantially as shown in FIG.

[0057] In some embodiments, the solvate form of Compound 1 is Form C prepared by vapor diffusion using isopropyl alcohol and has the following concentrations: 3.38, 6.85, 8.35, 10.95, 13.31, 13.84, 16.31, 16.65, 16.99, 17.23, 18.76, 19.33, 19.6, 20.13, 20.30, 20.80, 21.05, 21.86, 22.68, 23.13, 23.06, 23.04, 23.08, 23.09, 24.06, 24.08, 25.09, 26.07, 26.10, 27.09, 27.20, 27.21, 27.22, 27.23, 27.24, 27.25, 27.26, 27.27, 27.28, 27.29, 28.36, 28.35, 28.35, 28.45, 28.46, 28.47, 28.49, 29.26, 30.06, 30.05, 30.04, 30.02, 30.06, 30.08, 30.09, 30.10, 30.11, 30.12, 30.13, 30.14, 30.15, 30.16, 30.17, 30.18, 30.19, 30.20, 30.21, 30.22, 30.23, 0.6, 23.88, 24.3, 24.9, 25.64, 26.24, 26.88, 27.26, 28, 28.53, 29.53, 30.66, 31.64, 32.59, 32.95, 33.74, 34.78, 36.21, 36.94, 38.24, 40.17, 40.80, or 43.55 degrees 2θ, each ±0.2 degrees 2θ.

[0058] In some embodiments, the solvate form of Compound 1 is Form C prepared by vapor diffusion using isopropyl alcohol and has the following concentrations: 3.38, 6.85, 8.35, 10.95, 13.31, 13.84, 16.31, 16.65, 16.99, 17.23, 18.76, 19.33, 19.6, 20.13, 20.30, 20.80, 21.05, 21.86, 22.68, 23.13, 24.06, 25.05, 26.09, 27.04, 28.08, 29.09, 30.06, 31.07, 32.09, 33.10, 34.04, 35.06, 37.08, 38.09, 39.09, 40.06, 41.08, 42.09, 43.09, 44.09, 45.09, 46.09, 47.09, 48.09, 49.09, 50.09, 51.09, 52.09, 53.09, 54.09, 55.09, 56.09, 57.09, 58.09, 59.09, 60.09, 61.09, 62.09, 63.09, 64.09, 65.09, 66.09, 67.09, 68.09, 69.09, 70.09, 71.09, 72.09, 73.09, It has an X-ray powder diffraction (XRPD) pattern which includes all peaks at 23.6, 23.88, 24.3, 24.9, 25.64, 26.24, 26.88, 27.26, 28, 28.53, 29.53, 30.66, 31.64, 32.59, 32.95, 33.74, 34.78, 36.21, 36.94, 38.24, 40.17, 40.80, and 43.55 degrees 2θ, each ±0.2 degrees 2θ.

[0059] In some embodiments, Form C prepared by vapor diffusion using isopropyl alcohol, as analyzed by differential scanning calorimetry (DSC) thermogram, exhibits an endothermic peak at about 89.4° C. and a small endothermic peak at 134.2° C., as shown in FIG. 10.

[0060] In some embodiments, Form C prepared by vapor diffusion using isopropyl alcohol as analyzed by thermogravimetric analysis (TGA) exhibits dehydration with a weight loss of about 3.2% (w / w) from ambient temperature to about 200° C., as shown in FIG. 10.

[0061] Some embodiments provide a process for preparing Form C of Compound 1, comprising: (i) dissolving Compound 1 in isopropyl alcohol to form a saturated solution; (ii) filtering the mixture of step (i) into a vial that is placed within an outer vial containing an anti-solvent; (iii) stirring the mixture of the inner vial of step (ii) to precipitate a solid; and (iv) filtering the solid of step (iii) and drying the solid under vacuum at room temperature.

[0062] II. Pharmaceutical Compositions In some embodiments, the compounds and compositions of the present disclosure are administered to humans, human patients, or subjects.For the purposes of this disclosure, the phrase "active ingredient" typically refers to the compounds described herein.One aspect of the present disclosure provides a pharmaceutical composition comprising at least one pharma- ceutically acceptable carrier and the compounds of the present disclosure.

[0063] Although the description of pharmaceutical compositions provided herein is directed primarily to pharmaceutical compositions suitable for administration to humans, those skilled in the art will understand that such compositions are typically also suitable for administration to other animals, e.g., non-human animals, e.g., non-human mammals. Modifications to pharmaceutical compositions suitable for administration to humans to make them suitable for administration to a variety of animals are well understood, and typically, a skilled veterinary pharmacologist can design and / or implement such modifications, if necessary, with only routine experimentation. Subjects to which the pharmaceutical compositions are contemplated include, but are not limited to, humans and / or other primates, mammals, including commercially relevant mammals such as cows, pigs, horses, sheep, cats, dogs, mice, and / or rats, and / or birds, including commercially relevant birds such as poultry, chickens, ducks, geese, and / or turkeys.

[0064] The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. Typically, such preparation methods may include the steps of combining the active ingredient with an excipient and / or one or more other accessory ingredients and, if necessary and / or desirable, dividing, shaping and / or packaging the product into desired single or multiple dose units.

[0065] Pharmaceutical compositions according to the present disclosure may be prepared, packaged, and / or sold in bulk, as single dose units, and / or as a plurality of single dose units. As used herein, a "unit dose" refers to a discrete portion of a pharmaceutical composition comprising a predetermined amount of an active ingredient. The amount of the active ingredient is typically equal to the dose of the active ingredient that would be administered to a subject, and / or a convenient fraction of such a dose, such as one-half or one-third of such a dose.

[0066] The relative amounts of active ingredient, pharma- ceutically acceptable excipient, and / or any additional ingredients in a pharmaceutical composition according to the present disclosure will vary according to the identity, size, and / or condition of the subject being treated, and also according to the route by which the composition is administered. For example, the composition may comprise between 0.1% and 100%, e.g., between 0.5% and 50%, between 1% and 30%, between 5% and 80%, at least 80% (w / w) active ingredient.

[0067] The compounds of the present disclosure can be formulated with one or more excipients to (1) enhance stability, (2) enable sustained or extended release, (3) alter biodistribution, or (4) alter the release profile of the compound in vivo. Non-limiting examples of excipients include any solvent, dispersion medium, diluent, or other liquid vehicle, dispersion or suspension aid, surfactant, tonicity agent, thickening or emulsifying agent, and preservative. Thus, the formulations of the present disclosure may include one or more excipients in an amount that, together, enhances the stability of the compound.

[0068] Excipients Pharmaceutical formulations may include pharma- ceutically acceptable excipients, herein any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like, appropriate for the particular dosage form desired. Remington's The Science and Practice of Pharmacy, 21st Edition, AR Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006, incorporated herein by reference in its entirety), discloses various excipients used in formulating pharmaceutical compositions and known techniques for their preparation. Unless a conventional excipient vehicle is compatible with the substance or its derivatives, e.g., produces undesirable biological effects or adversely interacts with other components of the pharmaceutical composition, its use is considered within the scope of the present disclosure.

[0069] In some embodiments, a pharma- ceutically acceptable excipient is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, the excipient is approved for human and veterinary use. In some embodiments, the excipient is approved by the U.S. Food and Drug Administration (FDA). In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeia.

[0070] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents, disintegrating agents, binders, preservatives, buffers, lubricants, and / or oils, etc. Such excipients may optionally be included in the pharmaceutical composition.

[0071] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and / or combinations thereof.

[0072] Representative granulating and / or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponge, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinylpyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked sodium carboxymethylcellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (VEEGUM®), sodium lauryl sulfate, quaternary ammonium compounds, and / or combinations thereof.

[0073] Representative surfactants and / or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and VEEGUM® [magnesium aluminum silicate]), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, monostearate triacetate, ethylene glycol distearate, glyceryl monostearate, propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters, ... esters (e.g., polyoxyethylene sorbitan monolaurate [TWEEN® 20], polyoxyethylene sorbitan [TWEEN® 60], polyoxyethylene sorbitan monooleate [TWEEN® 80], sorbitan monopalmitate [SPAN® 40], sorbitan monostearate [SPAN® 60], sorbitan tristearate [SPAN® 65], glyceryl monooleate, sorbitan monooleate [SPAN® 80]), polyoxyethylene ... monolaurate [TWEEN® 60], polyoxyethylene sorbitan monooleate [TWEEN® 80], polyoxyethylene sorbitan monopalmitate [SPAN® 40], sorbitan monostearate [SPAN® 60], sorbitan tristearate [SPAN® 65], glyceryl monooleate, sorbitan monooleate [SPAN® 80]), polyoxyethylene esters (e.g., polyoxyethylene sorbitan monolaurate [TWEEN® 20], polyoxyethylene sorbitan monolaurate [TWEEN® 60], polyoxyethylene sorbitan monolaurate [TWEEN® 80]), polyoxyethylene esters (e.g., polyoxyethylene monostearate [MYRJ® 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Kolliphor® (SOLUTOL®), fatty acid esters, polyethylene glycol fatty acid esters (e.g., CREMOPHOR®), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether [BRIJ® 30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate,These include, but are not limited to, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, PLUORINC® F 68, POLOXAMER® 188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, Kolliphor SLS, and / or combinations thereof.

[0074] Exemplary binders include, but are not limited to, starches (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husk, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum®), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and / or combinations thereof.

[0075] Representative preservatives may include, but are not limited to, antioxidants, chelating agents, antibacterial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and / or other preservatives. Representative antioxidants include, but are not limited to, alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and / or sodium sulfate. Representative chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and / or trisodium edetate. Representative antibacterial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetide, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and / or thimerosal. Representative antifungal preservatives include, but are not limited to, butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and / or sorbic acid. Representative alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoates, and / or phenylethyl alcohol. Exemplary acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dihydroacetic acid, ascorbic acid, sorbic acid, and / or phytic acid.Other preservatives include tocopherol, tocopheryl acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfate, potassium metabisulfite, GLYDANT PLUS®, PHENONIP®, methylparaben, GERMALL® 115, GERMABEN® II, and NEOLONE. TM , KATHON TM , and / or EUXYL®.

[0076] Buffering agents include, but are not limited to, citrate buffer, acetate buffer, phosphate buffer, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, dibasic calcium phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixture, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixture, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixture, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and / or combinations thereof.

[0077] Representative lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and combinations thereof.

[0078] Representative oils include almond, apricot kernel, avocado, babassu, bergamot, blackcurrant seed, borage, cade, chamomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver oil, coffee, corn, cottonseed, emu, eucalyptus, evening primrose, fish, linseed, geraniol, gourd, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, Oils that may be used include, but are not limited to, lemon, lily of the valley, macadamia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasuna, savory, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, camellia, vetiver, walnut, and wheat germ oil. Representative oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and / or combinations thereof.

[0079] Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and / or perfuming agents can be present in the formulation, at the discretion of the formulator.

[0080] In some embodiments, the composition of the present disclosure comprises an AH crystal form of Compound 1 (e.g., Form A), a solvate form of Compound 1 (e.g., Form B and Form C), or a mixture thereof, optionally at least one carrier (e.g., but not limited to, lactose monohydrate), optionally at least one disintegrant (e.g., but not limited to, croscarmellose sodium), and / or optionally at least one lubricant (e.g., but not limited to, magnesium stearate). In some embodiments, the composition comprising Compound 1 is a nanomilled suspension or a spray-dried nanosuspension. The composition optionally comprises hypromellose (HPMC, e.g., HPMC 63), sodium lauryl sulfate (SLS, e.g., Kolliphor SLS), lactose monohydrate, croscarmellose, magnesium stearate, or a mixture thereof. The amount of Compound 1 in these compositions is between about 5% (w / w) and about 30% (w / w), such as about 5% (w / w), about 10% (w / w), about 15% (w / w), about 20% (w / w), about 25% (w / w), or about 30% (w / w). Non-limiting examples of nanosuspension and active granule compositions are shown below in Table 2. Non-limiting examples of active capsule compositions are shown below in Table 3. DL = drug loading.

[0081] Table 2. Nanosuspension and active granule composition [Table 5]

[0082] Table 3. Active Capsule Composition [Table 6]

[0083] Some embodiments of the present application provide pharmaceutical compositions comprising an effective amount of a compound, such as an AH crystal form of Compound 1 (e.g., Form A), a solvate form of Compound 1 (e.g., Form B and Form C), or a mixture thereof. In some embodiments, about 5% (w / w) to about 30% (w / w) of Compound 1 (e.g., Form A, Form B, or Form C) may be present in the pharmaceutical composition. In some embodiments, about 5% (w / w), 10% (w / w), 24% (w / w), or about 25% (w / w) of Compound 1 (e.g., Form A, Form B, or Form C) may be present in the pharmaceutical composition. The pharmaceutical composition may further comprise at least one of hypromellose (e.g., HPMC 603), sodium lauryl sulfate (e.g., Kolliphor SLS), or lactose monohydrate (e.g., 11SD), and, optionally, at least one of croscarmellose (e.g., Ac-di-Sol) or magnesium stearate (e.g., Ligamed MF-2-V). Examples of weight percentages of ingredients are shown in Table 4. In some embodiments, the pharmaceutical composition may be in the form of a capsule, e.g., a capsule containing 5 mg, 50 mg, or 100 mg of Compound 1 (e.g., Form A, Form B, or Form C).

[0084] Table 4. Weight percent of ingredients [Table 7]

[0085] III.How to use The mGluR5 receptor has emerged as a potential target for therapeutic benefit in many disease states. Based on the expression pattern and functional role of mGluR5, this receptor is an important target for drug discovery in many therapeutic indications. Evaluation of genetically engineered mice lacking mGluR5 and evaluation of compounds that modulate receptor function suggest that ligands that modulate mGluR5 receptor function may have therapeutic benefit in CNS and peripheral disease states. These include, but are not limited to, schizophrenia, cognitive impairment, Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia of Parkinson's disease, addiction, anxiety, depression, psychosis, epilepsy, fragile X syndrome, gastroesophageal reflux disease, migraine, pain, infectious or genetic prion diseases (such as, but not limited to, Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome (GSS) and fatal familial insomnia), tauopathies (such as frontotemporal dementia, corticobasal syndrome, Richardson's syndrome, parkinsonism, pure akinesia with gait immobility, rarely motor neuron symptoms or cerebellar ataxia), rare neurodegenerative diseases that resemble Parkinson's disease, and the like.

[0086] In some embodiments, a therapeutically effective amount of a compound or composition of the present disclosure can be used to treat CNS disorders and neurological or psychiatric disorders, such as schizophrenia, cognitive impairment, Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia in Parkinson's disease, addiction, anxiety, depression, psychosis, epilepsy, fragile X syndrome, gastroesophageal reflux disease, migraine headaches, pain, borderline personality disorder, bipolar disorder, or other neurological and / or psychiatric disorders associated with glutamate dysfunction.

[0087] In one embodiment, a method for treating Alzheimer's disease is provided, comprising administering a therapeutically effective amount of a compound or composition of the present disclosure to a patient in need thereof. The compound may be Compound 1. In some embodiments, a therapeutically effective amount of a compound or composition of the present disclosure is used to reverse synaptic density loss or synaptic loss in a subject in need thereof, such as loss of synaptic density in the cerebral cortex and hippocampus. In some embodiments, a therapeutically effective amount of a compound or composition of the present disclosure is used to reduce tau accumulation in a subject in need thereof.

[0088] In some embodiments, a method for preventing convulsions is provided, comprising administering to a patient in need thereof a therapeutically effective amount of a compound or composition of the present disclosure. The compound may be Compound 1.

[0089] In some embodiments, the compounds and compositions of the disclosure are used in the manufacture of a medicament for the treatment of CNS disorders and neurological or psychiatric disorders, such as schizophrenia, cognitive impairment, Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia in Parkinson's disease, addiction, anxiety, depression, psychosis, epilepsy, fragile X syndrome, gastroesophageal reflux disease, migraine headaches, pain, borderline personality disorder, bipolar disorder, or other neurological and / or psychiatric disorders associated with glutamate dysfunction.

[0090] In one embodiment, a compound or composition of the disclosure is used in the manufacture of a medicament for the treatment of Alzheimer's disease. The compound may be Compound 1.

[0091] Alzheimer's disease Despite millions of people suffering from it, there is still no disease-modifying treatment for Alzheimer's disease (AD). Without being bound by theory, the diagnosis of AD requires evidence of Aβ peptide accumulation. The main concomitant symptoms are the accumulation, phosphorylation and diffusion of misfolded tau (MAPT), and microglial and astroglial reactivity. Genetic evidence from dominant early-onset Alzheimer's disease mutations in APP and presenilin, Down's syndrome, and protective APP alleles all support Aβ as a causative agent. Biomarker studies have documented Aβ accumulation 20 years before symptoms appear, supporting an Aβ trigger. However, the temporal dissociation between Aβ accumulation and symptoms, as well as the clinical failure of multiple Aβ-lowering therapies, has widened the focus of AD pathophysiology. Genetic studies have implicated microglia in AD risk and progression, and the role of brain innate immunity in symptomatic AD is of increasing interest. For example, TREM2 signaling has multiple effects on Aβ-induced pathology.Furthermore, components of the classical complement system are directly involved in synapse elimination by phagocytosis in AD, although the basis for synapse selectivity remains unclear.

[0092] Synapse loss is important in the clinical progression of Alzheimer's disease, from the pre-symptomatic stage through mild cognitive impairment (MCI) to dementia. Synapse loss was initially documented ultrastructurally at autopsy, but can now be tracked indirectly with FDG-PET or directly with SV2A PET. PET tracers targeting synaptic vesicle glycoprotein 2A (SV2A), which is widely present in synaptic vesicles, allow for the quantification of synapse density by non-invasive scans, facilitating tracking of the progression of neurodegenerative diseases and drug development. To date, [ 11PET imaging with [C]UCB-J demonstrated that hippocampal synaptic density was significantly reduced in patients with Alzheimer's disease-related MCI compared to healthy controls, and also detected drug-induced reversal of synaptic loss in a mouse AD model (APPswe / PS1ΔE9, hereafter referred to as APP / PS1). 18 F]SynVesT-1 has a long half-life and high resolution, making it more likely to be applied clinically, and was able to distinguish differences in synaptic density between APP / PS1 mice and control littermates.

[0093] Regarding the biochemical events at impaired AD synapses, cellular prion protein (PrP C ) was identified as a high affinity receptor for Aβ oligomers (Aβo) through the only reported unbiased genome-wide expression cloning screen. C shows specific selectivity for oligomeric Aβ. In AD models, PrP C Binding of Aβo to PrP has been implicated in impaired synaptic plasticity, learning and memory deficits, and synapse loss. C The search for postsynaptic transmembrane proteins linking mGluR5 to intraneuronal signaling has revealed that the neuronal metabotropic glutamate receptor 5 (mGluR5) is a key link to Fyn and Pyk2 (PTK2B). Importantly, these kinases link to tau and are implicated in the risk of Alzheimer's disease. Genetic deletion and pharmacological inhibition studies have shown that reducing mGluR5 activity alleviates synaptic and memory impairments in multiple AD models. However, the molecular and cellular basis of the role of mGluR5 in AD synapse loss, including neuronal-glial interactions and complement system-synaptic phagocytosis, remains unclear.

[0094] Allosteric mGluR5 modulators have been classified as positive (PAMs), negative (NAMs), or silent (SAMs). PAMs mediate glutamate-induced, G protein-mediated Ca 2+NAMs enhance recruitment, inhibit and / or alter the potency of glutamate. Several NAMs mediate physiological glutamate signaling and Aβo-PrP signaling. C As a dose-limiting side effect, blockade of glutamate at mGluR5 by NAM impairs learning and memory independent of AD. Surprisingly, Compound 1 does not alter basal or glutamate signaling, but reduces PrP C Compound 1 inhibits mGluR5-mGluR5 interactions to prevent pathological Aβo signaling. Compound 1 significantly expands the therapeutic potential of using mGluR5 as a disease-modifying AD target.

[0095] In some embodiments, Compound 1 is used to modulate neuronal and glial gene expression, regulate neuroimmune interactions in AD synapse loss, restore synaptic density, and prevent synaptic localization of C1q without altering total C1q levels or overall gliosis.

[0096] In some embodiments, compound 1 increases synaptic density, slows loss of synaptic density, reduces loss of synaptic density, increases the levels of pre- and post-synaptic markers (SV2A and PSD-95), reverses loss of synaptic markers (SV2A and PSD-95), prevents seizures, and inhibits human transporters P-glycoprotein (P-gp), organic ion transporting polypeptide (OATP) 1B1, and OATP1B3. In some embodiments, compound 1 is used to modulate cerebrospinal fluid (CSF) or plasma biomarkers, such as, but not limited to, total tau protein levels, phosphorylated tau protein levels, SNAP-24, neurofilament light chain, GAP-43, synaptotagmin-1, α-synuclein (including phosphorylated forms), and neurogranin. In some embodiments, compound 1 is used to modulate complement activation biomarkers.

[0097] In some embodiments, patients administered Compound 1 exhibit changes in fluorodeoxyglucose (FDG) positron emission tomography (PET) imaging, synaptic vesicle glycoprotein 2A (SV2A) PET imaging, metabotropic glutamate receptor subtype 5 (mGluR5)-PET imaging, tau PET imaging, amyloid beta PET imaging, electroencephalogram (EEG) activity, functional magnetic resonance imaging, and / or volumetric magnetic resonance imaging.

[0098] In some embodiments, a patient is diagnosed with either amnesic mild cognitive impairment (aMCI) or mild dementia due to AD, based, for example, on the following: JPEG2025509988000013.jpg77169

[0099] Combination therapy In some embodiments, the present invention provides a method for treating the disease or disorder described herein, comprising administering the compound or composition of the present disclosure in combination with one or more additional active agents or therapies.Suitable pharmaceutical compounds or therapies that may be used in combination with the compound of the present disclosure include, but are not limited to, anti-amyloid immunotherapy, anti-tau immunotherapy, active agents aimed at reducing all forms of amyloid beta levels, active agents aimed at inhibiting the toxicity of amyloid beta oligomers, targeted treatment of microglial inflammation, acetylcholinesterase inhibitors, and NMDA receptor antagonists (e.g., but not limited to, donepezil and memantine).

[0100] The disclosed compounds or compositions and the additional active agent(s) may be administered simultaneously, sequentially, or in any order. The disclosed compounds or compositions and the additional active agent(s) may be administered in different doses, at different administration frequencies, or by different routes, whichever is appropriate.

[0101] IV. Dosage and Administration The present disclosure encompasses delivery of the compounds or compositions of the present disclosure for any therapeutic, prophylactic, pharmaceutical, diagnostic, or imaging use by any suitable route, taking into account possible advances in drug delivery science.

[0102] The compounds or compositions of the present disclosure may be administered by any route that produces a therapeutically effective result, including, but not limited to, oral, intravenous (intravenous), intrathecal (administration into the spinal or subarachnoid space to reach the cerebrospinal fluid), intraparenchymal (administration into the brain parenchyma), ear drops, nasal aerosol, and inhalation. In certain embodiments, the compounds or compositions may be administered in a manner that allows them to cross the blood-brain barrier, vascular barrier, or other epithelial barrier.

[0103] In some embodiments, the compounds or compositions of the present disclosure may be orally administered in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions and solutions. For tablets for oral administration, commonly used carriers include lactose and cornstarch. Lubricants, such as magnesium stearate, are also typically added. For oral administration in capsule form, useful diluents may include lactose and dried cornstarch. For oral administration of aqueous suspensions, the active ingredient may be mixed with emulsifying and suspending agents. If necessary, certain sweeteners and / or flavors and / or colorants may be added.

[0104] In some embodiments, the compounds or compositions of the present disclosure may be formulated for administration to the CNS by routes well known in the art, including, but not limited to, direct intraparenchymal administration, intrathecal administration, and intraventricular injection.

[0105] Dosage form The pharmaceutical compositions described herein can be formulated into dosage forms described herein, such as capsules, tablets, aqueous suspensions or solutions, topical, nasal, intratracheal, or injectable (e.g., intravenous, intraocular, intravitreal, intramuscular, intraventricular, intraperitoneal, subcutaneous). It will be understood that the total daily usage of the compositions of the present disclosure may be determined by the attending physician within the scope of medical judgment. The therapeutically effective, prophylactically effective, or suitable imaging dose level for a particular patient will depend on a variety of factors, including the disorder being treated and the severity of the disorder, the activity of the specific compound used, the specific composition used, the age, weight, general health, sex, and diet of the patient, the time of administration, the route of administration, the rate of excretion of the specific compound used, the duration of treatment, drugs used in combination or with the specific compound used, and similar factors well known in the medical art.

[0106] In some embodiments, compositions according to the present disclosure can be administered at a dose of from about 0.0001 mg / kg to about 100 mg / kg, from about 0.001 mg / kg to about 0.05 mg / kg, from about 0.005 mg / kg to about 0.05 mg / kg, from about 0.001 mg / kg to about 0.005 mg / kg, from about 0.05 mg / kg to about 0.5 mg / kg, from about 0.01 mg / kg to about 50 mg / kg, from about 0.1 mg / kg to about 40 mg / kg, from about 0.5 mg / kg to about 30 mg / kg, from about 0.01 mg / kg to about 10 mg / kg, from about 0.1 mg / kg to about 10 mg / kg, or from about 1 mg / kg to about 25 mg / kg, from about 25 mg / kg to about 50 mg / kg, from about 50 mg / kg to about 100 mg / kg of a subject's body weight for a therapeutic, diagnostic, prophylactic, or imaging effect. The desired dose may be administered once or multiple times daily at a dosage level sufficient to administer up to about 100 mg / kg, about 125 mg / kg, about 125 mg / kg to about 150 mg / kg, about 150 mg / kg to about 175 mg / kg, about 175 mg / kg to about 200 mg / kg, or about 200 mg / kg to about 250 mg / kg. The desired dose may be administered three times a day, twice a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks. In some embodiments, the desired dose may be administered using multiple administrations (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or more administrations). When multiple administrations are employed, there may be a split dose regimen, for example as described herein. In some embodiments, a compound or composition of the disclosure is administered by continuous infusion.

[0107] As used herein, "split dose" refers to the division of a single dose or total daily dose into two or more doses, e.g., two or more administrations of a single dose. As used herein, "single dose" refers to any therapeutic dose administered in one administration / one time / single route / single contact, i.e., a single administration event. As used herein, "total daily dose" refers to the amount administered or prescribed within a 24-hour period. It may be administered as a single dose.

[0108] The administration of the compounds or compositions of the present disclosure can be used as chronic or acute therapy.The amount of drug that may be combined with carriers to produce a single dosage form varies depending on the host treated and the particular mode of administration.A typical preparation contains about 5% to about 95% active compound (w / w).

[0109] If the patient's condition improves, a maintenance dose of the disclosed compound, composition, or combination may be administered as needed. Thereafter, if symptoms are alleviated to a desired level, the dosage and / or frequency of administration should be reduced symptomatically to a level sufficient to maintain the improved condition, and treatment should be terminated. However, patients may require intermittent treatment on a long-term basis in the event of a recurrence of disease symptoms.

[0110] As the skilled artisan will appreciate, there may be cases where lower or higher doses than those set forth above are required. The specific dosage and treatment regimen for a particular patient will depend on a variety of factors, including the activity of the particular compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, severity and course of infection, predisposition of the patient to infection, and the judgment of the treating physician.

[0111] In some embodiments, the subject is treated with about 10 mg, about 40 mg, about 70 mg, about 100 mg, about 150 mg, or about 200 mg of Compound 1. In some embodiments, the subject is treated once or twice daily. In a non-limiting example, the subject is treated with about 75 mg to about 125 mg of Compound 1 twice daily. In another non-limiting example, the subject is treated with at least 200 mg of Compound 1 once daily.

[0112] In some embodiments, the compound or composition of the present disclosure is administered by oral route in a capsule. In some embodiments, the capsule contains about 5 mg, about 25 mg, about 50 mg, about 75 mg, or about 100 mg of the compound. In some embodiments, the capsule further comprises lactose monohydrate, croscarmellose sodium, and / or magnesium stearate. The lactose monohydrate may be spray-dried. The croscarmellose sodium and magnesium stearate are in the extragranular form. In some embodiments, the concentration of the compound in the capsule is between about 1% (w / w) and about 50% (w / w), for example, between about 1% (w / w) and about 5% (w / w), between about 6% (w / w) and about 10% (w / w), between about 11% (w / w) and about 20% (w / w), between about 21% (w / w) and about 30% (w / w), between about 31% (w / w) and about 40% (w / w), or between about 41% (w / w) and about 50% (w / w). In one embodiment, the capsule contains about 5 mg of compound 1, and compound 1 has a concentration of about 5% (w / w) in the capsule. In other embodiments, the capsule contains about 50 mg or 100 mg of compound 1, and compound 1 has a concentration of about 25% (w / w) in the capsule.

[0113] In some embodiments, the capsules are stored at 25° C. / 60% relative humidity (RH). The compounds within the capsules are stable for at least 30 months under accelerated and long-term stability conditions, with the capsules being stable for at least 1 month.

[0114] V. Kits and Equipment The present disclosure also provides various kits and devices for conveniently and / or effectively carrying out the methods of the present disclosure. Typically, the kits include components in amounts and / or numbers sufficient to allow a user to administer multiple treatments to a subject and / or perform multiple experiments.

[0115] In one embodiment, the disclosure provides a kit for treating a CNS disorder comprising a compound or composition of the disclosure, optionally in combination with any other active agent.

[0116] The kit may further include packaging and instructions for preparing the formulation composition, and / or a delivery agent. The delivery agent may include saline, a buffer, or any of the delivery agents disclosed herein. The amount of each component may be varied to allow for consistent and reproducible high concentration saline or simple buffer formulations. Components may also be varied to increase the stability of the compound in the buffer over time and / or under various conditions.

[0117] The present disclosure provides devices that may incorporate the compounds or compositions of the present disclosure. These devices contain a stable formulation that can be immediately administered to a subject in need thereof, such as a human patient. In some embodiments, the subject has a CNS disorder.

[0118] Non-limiting examples include pumps, catheters, needles, transdermal patches, pressurized olfactory delivery devices, iontophoresis devices, and multi-layer microfluidic devices. These devices may be used to deliver the compounds or compositions of the present disclosure according to single dose, multiple dose, or split dose regimens. These devices may be used to deliver the compounds or compositions of the present disclosure to biological tissue, intradermally, subcutaneously, or intramuscularly.

[0119] VI.Definitions For convenience, the meaning of certain terms and phrases used in the specification, examples, and appended claims are provided below. If there is an apparent discrepancy between the usage of a term in other parts of the specification and the definition provided in this section, the definition in this section shall control.

[0120] The abbreviations used herein have their usual meaning in the scientific and technical fields. Chemical elements are identified according to the Periodic Table of the Elements, CAS edition, Handbook of Chemistry and Physics, 75th edition. In addition, the general principles of organic chemistry are described in M. Loudon, Organic Chemistry, 5th edition, Roberts and Company, Greenwood Village, Colo.: 2009, and M.B. Smith, March's Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 7th edition, John Wiley & Sons, Hoboken: 2013, the entire contents of which are incorporated herein by reference.

[0121] As used herein, the term "about" means ±10% of the stated value.

[0122] As used herein, the term "compound" is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted.

[0123] The term "substantially purified" refers to a state free from other dissimilar compounds and impurities normally associated with the crystalline forms of the present invention in their natural state, and thus a "substantially purified" crystalline form is at least 95% by mass by weight of a given sample.

[0124] As used herein, the term "subject" or "patient" refers to any living organism to which a particle may be administered, e.g., for experimental, therapeutic, diagnostic, and / or prophylactic purposes. Exemplary subjects include animals (e.g., mammals such as mice, rats, rabbits, guinea pigs, cows, pigs, sheep, horses, dogs, cats, hamsters, llamas, non-human primates, and humans).

[0125] As used herein, the terms "treatment" or "prevention" include preventing the onset of a disease, disorder, and / or condition in an animal that is predisposed to the disease, disorder, and / or condition but has not yet been diagnosed with the disease, disorder, or condition, inhibiting the disease, disorder, or condition, e.g., preventing its progression, and relieving the disease, disorder, or condition, e.g., causing regression of the disease, disorder, and / or condition. Treating a disease, disorder, or condition can also include improving at least one symptom of a particular disease, disorder, or condition even if the underlying pathophysiology is not affected, e.g., treating a subject for pain by administering a painkiller, but the painkiller does not treat the cause of the pain.

[0126] As used herein, the terms "management" or "maintenance" can refer to reducing the symptoms of a disease, reducing the severity of the symptoms of a disease, or preventing the symptoms of a disease from worsening.

[0127] The term "therapeutic effect" is scientifically accepted and refers to a local or systemic effect produced in animals, particularly mammals, and more particularly humans, by a pharmacologically active substance. The term therefore refers to any substance intended for use in the diagnosis, cure, mitigation, treatment or prevention of a disease, disorder or condition in an animal, such as a human, in enhancing a desired physical or mental development and condition.

[0128] The term "modulation" is scientifically accepted and refers to the upregulation (i.e., activation or stimulation), downregulation (i.e., inhibition or suppression) of a response, or a combination or dissociation of the two. Modulation is typically compared to a baseline or reference value, which may be internal or external to the treated entity.

[0129] The terms "sufficient" and "effective", as used interchangeably herein, refer to an amount (e.g., mass, volume, dose, concentration, and / or duration) necessary to achieve one or more desired results. A "therapeutically effective amount" is the minimum concentration required to cause a measurable improvement or prevention of at least one symptom or a particular condition or disorder, or to cause a measurable life-prolonging effect, or typically to improve the quality of life of a patient. Thus, a therapeutically effective amount depends on the particular biologically active molecule and the particular condition or disorder to be treated. Therapeutically effective amounts of many therapeutic agents, such as antibodies, are well known in the art. Therapeutically effective amounts of the compounds and compositions described herein, for example for the treatment of a particular disorder, may be determined by techniques within the skill of a skilled artisan, e.g., a physician.

[0130] The terms "bioactive agent" and "active agent," as used interchangeably herein, include, but are not limited to, physiologically or pharmacologically active agents that act locally or systemically in the body. A bioactive agent refers to an agent used for therapy (e.g., therapeutic agents), prevention (e.g., prophylactic agents), diagnosis (e.g., diagnostic agents), treatment or mitigation of disease or illness, an agent that affects the structure or function of the body, or a prodrug that becomes biologically active or more active after being placed in a defined physiological environment.

[0131] As used herein, "pharmacologically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, within the scope of sound medical judgment, in accordance with guidelines from agencies such as the U.S. Food and Drug Administration (FDA), and are commensurate with a reasonable benefit / risk ratio. As used herein, "pharmacologically acceptable carriers" refers to all components of a pharmaceutical formulation that facilitate delivery of the composition in vivo. Pharmaceutically acceptable carriers include, but are not limited to, diluents, preservatives, binders, lubricants, disintegrants, swelling agents, fillers, stabilizers, and combinations thereof.

[0132] The term "pharmaceutically acceptable salt" refers to salts of acidic or basic groups that may be present in compounds used in the present compositions.

[0133] The term "substantially purified" refers to a purity of at least 90%, such as about 90%, about 95%, or about 99%, relative to other forms of the compound.

[0134] The details of one or more embodiments of the present disclosure are described in the accompanying description below. Although materials and methods similar or equivalent to those described herein can be used to practice or test the present disclosure, the preferred materials and methods are described herein. Other features, objects, and advantages of the present disclosure will become apparent from the description. In the description, the singular form includes the plural form unless the context clearly indicates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present disclosure belongs. In case of conflict, the present specification will control.

[0135] The present disclosure is further illustrated by the following non-limiting examples.

[0136] Working Example Example 1. Polymorph screening and characterization Crystallization Development The polymorphism testing protocol used a selection of solvents that were first used to evaluate the solubility of Compound 1. Four types of tests were then performed: temperature cycling, evaporation, long-term slurry, and vapor diffusion. The number of solvents, amount of solvent, and amount of sample used in each test were determined based on the solubility values ​​and the type of test.

[0137] The approximate solubility (visual) of compound 1 was evaluated in a list of solvents selected based on class diversity, including solvent / water mixtures (Table 5). Compound 1 (5.0±0.2 mg) was weighed into a vial and a known amount of solvent was added slowly at 20° C. and stirred until a clear solution was visually observed. The final solution was stirred overnight at room temperature to evaluate the recrystallization phenomenon.

[0138] Table 5. Estimated solubility of compound 1 [Table 8]

[0139] Typically, this compound exhibited moderate / high solubility in most of the solvents screened. Only heptane and water acted as antisolvents. Due to the high solubility, the solvents for the following experiments were selected based on the solvent properties (e.g., boiling point, miscibility) and solubility.

[0140] A total of 38 samples were prepared. Experiments included in the protocol were temperature cycling, evaporation, long-term slurry, and vapor diffusion experiments.

[0141] Temperature Cycling Slurries of compound 1 were prepared in eight solvent systems with solubilities <30 mg / mL. The slurries were subjected to six temperature cycles under magnetic stirring (650 rpm) in an Integrity workstation. Each cycle included a heating / cooling ramp at a rate of 2 °C / min and hold times at 20 °C and 40 °C for 110 min. The presence of solids under high temperature conditions was confirmed by visual inspection. Solid samples were isolated by filtration through a 20 μm PTFE filter syringe and dried under vacuum at 20 °C for approximately 2 h before being subjected to XRPD analysis.

[0142] Evaporation experiment Twelve organic solvents with boiling points lower than 110 °C and solubilities greater than 15 mg / mL were selected for evaporation experiments.

[0143] Near-saturated solutions of compound 1 were prepared in the selected solvent system and stirred for approximately 1 h at room temperature to maximize solubility equilibrium. The mixture was filtered through a 0.45 μm PTFE filter into a new vial. The resulting clear solution was evaporated in a dry box under inert atmosphere (e.g., nitrogen gas) (vials were open and no stirring was performed).

[0144] Long-term slurry Slurries of compound 1 were prepared in eight solvent systems selected for solubility < 30 mg / mL. The slurries were placed in a thermostatic bath at 20°C and stirred for 2 weeks.

[0145] Solvate form B (i.e., solvate S1) was prepared using an evaporation experiment. A nearly saturated solution of compound 1 was prepared in isopropyl alcohol and stirred at room temperature for approximately 1 hour to maximize the solubility equilibrium. The mixture was filtered through a 0.45 pm PTFE filter into a new vial (the vial was open and no stirring was performed). Within one week, the isopropyl alcohol had almost completely evaporated. The residue was dried in a vacuum oven at 20 °C for 1 hour.

[0146] Solvate form C (i.e., solvate S2) was prepared using a long-term slurry in isopropyl alcohol. The slurry was placed in a thermostatic bath at 20° C. and stirred for 2 weeks. The solid sample was isolated by filtration through a 20 pm PTFE filter syringe and dried under vacuum for approximately 2 hours.

[0147] Solvate form C (i.e., solvate S2) was also prepared by vapor diffusion method using isopropyl alcohol and pentane as anti-solvents. A concentrated solution of compound 1 was prepared in isopropyl alcohol and stirred at room temperature for approximately 1 hour to maximize the solubility equilibrium. The mixture was filtered through a 0.45 pm PTFE filter into a new vial and placed uncapped into the outer vial containing pentane. The system was sealed, stored at room temperature, stirred, and periodically checked for solid formation. Form C solids formed only when the mixture was stirred. The solids were isolated by removing the supernatant liquid and dried in a vacuum oven at room temperature for at least 30 minutes.

[0148] The results are summarized in the table below.

[0149] Table 6. Results of various solvents in different processes [Table 9]

[0150] Further testing showed that Form A was anhydrous (AH) and could be isolated using a mixture of EtOAc and heptane to give crystalline material in high yield and purity.

[0151] Solubility data were generated applying the following protocol: Compound 1 slurries were prepared and equilibrated at 20 °C with stirring for approximately 18 hours. To measure the concentration in the supernatant, approximately 0.1 mL of the slurry was taken with a syringe, filtered, weighed, diluted with MeCN, and injected into the HPLC. The slurry was heated at 45 °C for approximately 3 hours, after which sampling was repeated to obtain solubility data under elevated temperature conditions. The samples were cooled to 20 °C, filtered through a 20-μm PTFE filter to separate the solid residue, dried under vacuum at 40 °C, and subjected to X-ray diffraction analysis. The peak area of ​​compound 1 for each sample was recorded, and the solubility data was calculated using the known response factor (245 nm, 1460 mAU*sec / mg / mL, Table 7). The data was integrated and the solubility was visualized on a chart as a function of heptane volume percent. Compound 1 was highly soluble in pure EtOAc and in the presence of relatively high percentages of heptane (10-20). The solubility slowly decreased with increasing amounts of heptane. All solid residues exhibited a physical morphology consistent with Form A.

[0152] Table 7. Solubility data for compound 1 [Table 10]

[0153] Based on the solubility data, the metastable zone width of EtOAc / heptane (80:20, 70:30, 60:40) was examined to identify suitable operating conditions / seeding points.

[0154] An Electrothermal Integrity instrument coupled with an IR turbidity probe was used to generate the metastable zone width. Samples containing different amounts of material were weighed in vials and diluted with 1 mL of the appropriate solvent mixture. Screening concentrations, estimated to correspond to approximately 1 volume, taking into account the contribution of solids, ranged from 115 to 215 mg / mL. The step / plateau temperature program was set with a heating rate of 0.1 °C / min, a step of 1 °C, and a plateau time of 1 min. Step-up heating was set to 78 °C (the highest temperature allowed by the lowest boiling solvent component) and step-down cooling was applied with the same parameters. The vials were monitored with a turbidity probe to measure the elution temperature (overheat slope) and autocrystallization temperature (overcool slope) at each concentration.

[0155] All solvent compositions showed a regular dissolution profile that matched previously collected solubility data. Selected samples were isolated by filtration and analyzed by XRPD, which confirmed the consistency with Form A. Overall, the 80:20 EtOAc / heptane ratio seemed to be suitable for dissolution, while the composition 70:30 was preferred as a seeding point. Indeed, the latter provided a practical concentration range where the initial solution could be easily driven towards the supersaturated region. On the other hand, the MSZW at the 60:40 solvent ratio was judged to be too narrow and risky from a working point of view.

[0156] Characterization XRPD XRPD spectra were collected in transmission mode on a Panalytical X'Pert Pro or Empyrean instrument equipped with an X'Celerator detector using standard methods. Samples were irradiated with Cu Kα X-rays. Data were evaluated using Panalytical Data Viewer software. Details of the standard screening data collection method are as follows: A representative XRPD graph of AH Form A is shown in Figure 3. A representative XRPD graph of Form B is shown in Figure 5. Representative XRPD graphs of Form C are shown in Figures 7 and 9. Start position [°2θ] 2.0104 End position [°2θ] 44.9864 Step size [°2θ] 0.0170 Scan step time [s] 59.6900 Scan Type Continuous PSD mode scan PSD length [°2θ] 2.12 Offset [°2θ] 0.0000 Divergence slit type: fixed Divergence slit size [°] 0.4354 Sample length [mm] 10.00 Measurement temperature [℃] 25.00 Anode material Cu Intended Wavelength Type K-Alpha K-Alpha1[Å] 1.54060 K-Alpha2[Å] 1.54443 K-Beta[Å] 1.39225 K-A2 / K-A1 ratio 0.50000 Generator settings: 40mA, 40kV Diffractometer type 0000000011016732 Diffractometer Number 0 Angle meter radius [mm] 240.00 Distance Focal-Divergence Slit [mm] 100.00 Incident beam monochromator No Rotation Yes

[0157] TGA TGA analysis was performed on a TA Q5000 instrument. Details of the TGA method are listed below. A representative graph of AH Form A is shown in Figure 4. A representative graph of Form B is shown in Figure 6. Representative graphs of Form C are shown in Figures 8 and 10. JPEG2025509988000017.jpg50169

[0158] DSC DSC analysis was performed on a TA Q5000 instrument. Details of the DSC method are listed below. A representative graph for AH Form A is shown in Figure 4. A representative graph for Form B is shown in Figure 6. Representative graphs for Form C are shown in Figures 8 and 10. JPEG2025509988000018.jpg50169

[0159] Example 2. Crossover pharmacokinetic study of intravenous and oral administration in male CD rats This study assessed the PK of Compound 1 AH Form A in male Sprague Dawley rats (3 / group) following a single IV dose of 1 mg / kg in 2.5% N-methyl-2-pyrrolidone (NMP) + 1% Solutol HS15 or oral (PO) dosing of 3 formulations at 5 and 50 mg / kg to the same animal in a crossover fashion with a 7-day recovery period. For PO dosing, the compound was formulated as a nanomilled suspension (Formulation A) or two different solutions (Formulations B or C).

[0160] Serial plasma samples were collected at intervals up to 24 hours after dose administration, and the concentration of Compound 1 in each plasma sample was measured by LC-MS / MS. Pharmacokinetic parameters were measured and are shown in Table 8.

[0161] Table 8. Plasma pharmacokinetic parameters of Compound 1 after single intravenous and oral administration to male rats [Table 11] NA-Not applicable. 1 -AUC reported for IV dose and used to determine F% inf . A - Nanomilled suspension (10% (w / w) Compound 1) in 2% (w / w) HPMC 603 and 0.5% (w / w) SLS in MilliQ water. B - Water, 40% PEG 400, 20% Transcutol, 10% Vitamin E TPGS, 0.5% Poloxamer 407 C-in water, 5% DMSO, 0.5% HPMC

[0162] After each PO dose, an initial plasma peak occurred 0.5 to 2 hours after dosing, levels then declined, and a second, higher peak occurred 6 to 8 hours after dosing. inf and AUC of PO last The F% calculated from was high for all formulations.

[0163] Formulation A provided PO exposure (AUC 0.6-fold and 1.3-fold higher than that of reference formulation C at 5 and 50 mg / kg, respectively. last ) was shown.

[0164] Formulation B provided PO exposure (AUC last ) was shown.

[0165] At 50 mg / kg, Compound 1 (AUC last , 168000 ng.h / mL) and had the highest C max (8847 ng / mL) was observed in the nanosuspension formulation (Form A), which was used in a 28-day toxicity study in rats and monkeys.

[0166] After IV administration, T 1 / 2 is 4.49 hours, V dss was 2.23 L / kg, and CL was 5.98 mL / min / kg.

[0167] Example 3. Methods for formulating compounds 1. Nanosuspension composition This study was conducted to select a suitable nanosuspension composition for the AH Form A of Compound 1. Three qualitatively different vehicles were prepared at a 200 g scale each to verify the feasibility and stability of nanomilled suspensions in these media.

[0168] The vehicle was prepared in advance by solubilizing the appropriate amount of polymer and surfactant in purified water (MilliQ®). Complete solubilization of the material was achieved by magnetic stirring. Compound 1 Form A was then added to the prepared vehicle and suspended (a uniform suspension was obtained within about 15 minutes for all compositions).

[0169] The list of materials is given in Table 9. The qualitative and quantitative formulation compositions are given in Table 10.

[0170] Table 9. List of substances [Table 12]

[0171] Table 10. Qualitative composition investigated [Table 13]

[0172] Aiming to investigate the feasibility of the nanogrinding process, a small-scale Dynomill (Dynomill RL) was used at this stage. After assembly, the milling chamber was filled to 65% of the total volume (50 mL) with grinding media beads (equivalent to 32.5 mL of 0.4 mm YTZ grinding beads, approximately 120 g). The produced nanosuspensions were characterized by particle size distribution (PSD) by a Malvern Mastersizer (LLS). Particle size distribution by Malvern Zeta sizer (Z-sizer), Zeta potential, X-ray diffraction (XRPD), Assay / Impurities by HPLC, and Homogeneity by HPLC.

[0173] Based on the physical and chemical characterization of the nanomilled composition, Compositions 2 and 3 reached the target size after 60 min of milling and showed similar PSD characteristics (unimodal distribution), whereas composition 1 showed a bimodal distribution due to the presence of particles larger than 1 μm. - Compositions 2 and 3 showed better Z potential values ​​consistent with typical stable nanosuspension products (values ​​below -25 mV). -XRPD data showed that the nanomilling process did not affect the crystal form in all three compositions. It can be concluded that:

[0174] Stability studies (under refrigerated conditions only) were performed on all three nanosuspension formulations. The results of the 7-day stability study focused on the appearance of the formulation, mixing method and sampling points.

[0175] Unofficial Stability: 7 days The three nanosuspensions produced were stored in a refrigerator for 7 days and then characterized. Particle size distribution by laser light scattering using a Malvern Mastersizer; Z potential measurement using Malvern Zetasizer X-ray pattern diffraction, and Homogeneity and impurities by HPLC.

[0176] Based on the physical and chemical characterization results, Composition 1 was determined to be less stable and less homogeneous.

[0177] Unofficial Stability: 14 days The stability of compositions 2 and 3 was evaluated after 14 days of refrigerated storage. The following tests were performed: Particle size distribution measurement by laser light scattering using a Malvern Mastersizer Zeta potential measurement by Malvern Zetasizer, X-ray pattern diffraction, and homogeneity and impurities by HPLC.

[0178] Based on the physical and chemical characterization results, both compositions 2 and 3 can be considered stable for 14 days under refrigerated conditions. However, composition 3 showed more favorable Z potential values.

[0179] Based on this consideration, the following activities were carried out using the following compositions:

[0180] Table 11. Composition to be implemented [Table 14]

[0181] 2. Oral formulation development

[0182] Table 12. List of substances [Table 15]

[0183] The prepared nanosuspension was divided into sublots (four portions) and used to study top spray granulation. Two types of water-soluble fillers / carriers were tested. -Lactose monohydrate, Supertab 11 SD (spray-dried grade) -Mannitol, Pearlitol 200 SD (spray-dried grade)

[0184] In order to inhibit crystal growth by the compound-carrier matrix, small amounts of the aforementioned substances were dissolved in a portion of the nanomilled suspension to be sprayed (1:1 ratio with Compound 1). Four top spray granulations (two carriers, two drug loadings) were carried out as follows: - 330g of nanomilled material (+ lactose 11 SD or mannitol 200 SD as bulking agent) was sprayed onto 550g of the selected carrier to give approximately 5% granule DL, a drug loading suitable for preparing a 5mg capsule strength. - 550g of nanomilled material (+ lactose 11 SD or mannitol 200 SD as bulking agent) was sprayed onto 100g of the selected carrier to give approximately 20% granule DL, a drug loading suitable for preparing 50mg and 100mg capsule strengths.

[0185] Table 13 below shows the composition of the nanosuspension diluted with bulking agent.

[0186] Table 13. Composition of diluted nanosuspensions [Table 16]

[0187] After the bulking agent was added and dissolved in a portion of the nanosuspension, the carrier was added to the granulator bowl and warmed before starting the top spray granulation. Tables 14 and 15 below show the composition of the four granules prepared.

[0188] Table 14. Theoretical granule composition using mannitol [Table 17]

[0189] Table 15. Theoretical granule composition with lactose [Table 18]

[0190] Four top spray granulations were performed to investigate the process feasibility and to produce sufficient material for subsequent capsule preparation. The resulting granulations (5% and 20% DL) appeared visually different. At 20% DL, coarse agglomerates were observed in both lactose and mannitol-based granulations, whereas at 5% DL, very fine agglomerates were observed.

[0191] As a general observation for the following manufacturing batches, these feasibility studies were performed at suboptimal batch sizes, especially for the 20% DL (due to the low amount of carrier), but slight adhesion of fines to the granulator walls was observed at both drug loadings. Adjustments to batch size and feed rate were then made to the next batch.

[0192] The four types of granules prepared were characterized for loss on drying (LOD) (IPC), particle size distribution (PSD), X-ray pattern diffraction (XRPD), particle size as reconstituted, XRPD as reconstituted, bulk and tapped density, granule uniformity and impurity profile.

[0193] From a processability point of view, the lactose composition has superior properties compared to mannitol (powder mixing motion in the granulator chamber). Furthermore, granules of 21% DL lactose were the only composition that could be reconstituted into the input nanosuspension PSD.

[0194] Taking these considerations into account, lactose has advantages as a carrier.

[0195] Exogenous granule excipients were added to the four granule batches prior to manual capsule filling. At this stage, 4% (w / w) disintegrants (croscarmellose, Ac-di-Sol) and 1% (w / w) lubricant (magnesium stearate) were added to each granule batch.

[0196] The equipment used to mix the granules and extragranular material was a low shear mixer (Pharmatec) with appropriate bowl size (1 L bowl for 20% (w / w) granules and 2 L bowl for 5% (w / w) granules).

[0197] The disintegrant and lubricant were each sieved through a 500 micron screen and added directly to the bowl containing the granules and mixed for 10 minutes at 17 rpm and 3 minutes at 17 rpm.

[0198] For each carrier used, two types of drug loading were prepared. Specifically: -4.62% of the final blend was used to prepare 5 mg capsules. -20.00% of the final blend was used to prepare 50 mg and 100 mg capsules.

[0199] The final formulations are listed in Table 16 in % (w / w).

[0200] Table 16. Final formulation composition [Table 19]

[0201] Consideration of bulking agents 1,800 g of nano-milled material (without bulking agent) was sprayed onto 500 g of lactose 11 SD (carrier) to obtain about 24% granule DL. Table 17 below shows the composition of the prepared granules.

[0202] Table 17. Granule composition without filler [Table 20]

[0203] From a processability standpoint, a larger amount of carrier (500 g) suited the mill bowl capacity and allowed for proper movement of the carrier during heating and granulation. After preheating the mill and carrier, spraying was started at a very low feed rate (approximately 2 g / min) and then increased to approximately 12 g / min during the process while maintaining the product temperature at 28-35°C. The granules produced were fine in appearance but visually granular, with a final yield of 92.14% (669 g granules).

[0204] Particle size distribution (by LLS and sieve analysis) Concerning PSD, the batch shows a tight monodisperse mode particle size distribution similar to the previous example, but a smaller x90 is observed (340 microns versus 500 microns in the feasibility test). The values ​​detected by laser light scattering are in agreement with the results of the sieve analysis.

[0205] A representative PSD of the nanosuspension is shown in Table 18.

[0206] Table 18. PSD characteristics of granule compositions. Values ​​shown are the average of three analyses. [Table 21]

[0207] X-ray diffraction pattern (XRPD) XRPD of 20% DL granules (light green trace) shows the presence of compound 1.

[0208] Particle size distribution upon reconstitution The produced granules were tested for reconstitution to the nano-level. The reconstitution rate is lower than the lactose compositions produced in the previous examples. The higher rate previously observed is due to the presence of a bulking agent acting as a "stabilizer" and "steric hindrance" between the compound-compound particles, preventing the formation of agglomerates. Despite this, a portion of the compound particles was freely reconstituted into the same nano-range input nanosuspension.

[0209] Bulk and tapped density upon reconstitution. This composition exhibited improved flow properties compared to the previous examples, as confirmed by the "Hausner Ratio" index.

[0210] Granule uniformity The uniformity of the granules was checked. Six samples were taken from the bulk. As shown in the table below, a good potency of an average of 100.76% of the theoretical drug loading was obtained for the granules, in contrast to the low average value (87% (w / w)) found in the feasibility study, confirming the impact of inadequate batch size at this stage. A slightly higher RSD was observed, but within the acceptable range (RSD < 5%).

[0211] No impurities were detected (less than 0.05%).

[0212] Capsule filling for dissolution evaluation To understand whether the absence of a bulking agent affects the release of the compound, dissolution studies were performed on six manually filled capsules prepared with 100 mg strength granules. The dissolution studies confirmed that the absence of a bulking agent improved the release of the compound. Although there was still a fairly high variability between the individual capsules (as previously observed), all samples achieved >90% release after 60 minutes.

[0213] Example 4: In vitro testing In vitro inhibition of human transporters P-gp, OATP1B1, OATP1B3 and BSEP Compound 1, Form A, was evaluated as a potential inhibitor of the human transporters P-glycoprotein (P-gp), organic ion transporting polypeptide (OATP) 1B1, and OATP1B3 in various in vitro cellular test systems, and also in BSEP in membrane vesicles.

[0214] The ability of the in vitro test system to identify transporter inhibitors was confirmed by a reduction in the transporter-mediated activity of the probe substrate in the presence of a positive control inhibitor, determined from parallel incubations with the test compound.

[0215] Under the current assay conditions, compound 1 form A was an inhibitor of probe substrate transport via P-gp, OATP1B1, OATP1B3 and BSEP with IC50 values ​​of 27.7, 6.74, 32.0 and 13.9 μM, respectively.

[0216] Supersomes TM In vitro CYP450 reaction phenotyping assay with overexpressed human CYP enzymes The CYPP450 enzymes involved in the phase I metabolism of Form A of Compound 1 were investigated in vitro. Form A of Compound 1 was metabolized by CYP1A2, CYP2D6, and CYP3A4 with T values ​​of 64, 14, and 176 minutes, and CLint of 0.261, 0.984, and 0.0786 μL / min / pmol CYP, respectively. No evidence of metabolism of Compound 1 by CYP2B6, CYP2C8, CYP2C9, and CYP2C19 was observed.

[0217] The predicted contribution of each CYP to the metabolism of compound 1 form A was assessed using the factor relative activity method. These calculations indicated that CYP1A2, CYP3A4, and CYP2D6 were responsible for 43.1%, 38.9%, and 18% of the total metabolism, respectively.

[0218] In vitro CYP inhibition of CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 in human liver microsomes In this study, the potential inhibitory effects of Form A of Compound 1 on CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 were evaluated. Known inhibitors of each CYP were used as positive controls.

[0219] The results showed that compound 1 form A inhibited several CYP enzymes with IC50s of 1.95, 6.17, 12.4, 52.1, and 55.7 μM against CYP1A2, CYP2C19, CYP2C9, CYP2C8, and CYP3A4 (atorvastatin alone), respectively.

[0220] No inhibition of CYP2D6 or CYP3A4 was observed when midazolam or nifedipine was used as substrates.

[0221] In vitro metabolite-dependent inhibition studies of CYP2D6 and CYP3A4 in human liver microsomes. In this study, we evaluated the metabolically-dependent inhibitory potential of Compound 1, Form A, against CYP3A4 and CYP2D6 in human liver microsomes. Troleandomycin and paroxetine, known metabolically-dependent inhibitors of CYP3A4 and CYP2D6, respectively, were used as positive controls.

[0222] The overall results indicated that Compound 1 Form A is not a metabolism-dependent inhibitor of CYP3A4 and CYP2D6.

[0223] Activation of PXR and AhR receptors by Form A of Compound 1 The objective of this study was to determine the ability of Compound 1, Form A, to activate the human and rat pregnane X receptor (PXR) and the human and rat aryl hydrocarbon receptor (AhR). Activation of the PXR is a marker of induction of CYP3A4 / 5 and CYP2C6, and activation of the AhR is a marker of induction of CYP1A.

[0224] Compound 1 activated the PXR receptor in humans with a mean maximal activation fold of 4.58 (29.6% of the positive control) and a calculated F2 value of 5.05 μM. The mean activity in rats was 4.70-fold at 30 μM.

[0225] At 30 μM, compound 1 activated the AhR receptor with a mean fold maximum activation of 1.78 in humans (1.11% of the positive control) and 1.42 in rats (0.946% of the positive control), suggesting that compound 1 is a potent inducer of CYP3A4 and CYP2C6.

[0226] Example 5. In vivo testing Compound 1 (e.g., Form A) inhibited the abnormal synaptic signaling that causes Alzheimer's disease while preserving physiological Glu activity. Studies have shown that orally administered Compound 1 inhibited the synaptic signaling that causes Alzheimer's disease at doses 250-fold lower than the side effect level in rodents and non-human primates. 18 F]FPEB effectively occupies mGluR5 sites in the brain as visualized by PET. In aged transgenic and double knock-in mouse models of Alzheimer's disease, [ 18SV2A PET imaging with [F]SynVesT-1 revealed a decrease in synaptic density in the cortex and hippocampus, which was fully reversed by treatment with compound 1. This disease-modifying effect persisted after drug administration was discontinued. Tau accumulation in double knock-in mice was also reduced by compound 1 treatment. Single-nucleus transcriptomics showed that compound 1 treatment normalized expression patterns to a much greater extent in neurons than in glial cells. Although production of the microglial mediator C1q was not altered by compound 1, synaptic localization and uptake of C1q, which is dependent on Alzheimer's disease-related genes, was inhibited. Thus, selective modulation of mGluR5 reversed changes in neuronal gene expression to protect synapses from damage by microglial mediators.

[0227] [ 18 F]FPEB substitution test mGluR5 PET ligand [ 18 Receptor occupancy was assessed in mice using [F]FPEB. Mice were orally dosed twice daily with different doses of compound 1. Available mGluR5 sites in the brain were assessed in anesthetized mice both at peak 1 h after a single dose and at trough after 7 days of twice daily (bid) oral dosing. A dose of 3.75 mg / kg, bid, maintained >90% receptor occupancy throughout the dosing period. This was consistent with a free trough drug level of 15 nM and [F]FPEB from mGluR5. 3 This corresponds to an inhibition constant (Ki) of 0.6 nM for compound 1 against [H]-MPEPy displacement. A dose of 3.75 mg / kg bid maintains free drug at 25-fold the Ki. As shown in Figure 1, compound 1 inhibited [H]-MPEPy displacement in mouse brain. 18 F]FPEB uptake was inhibited. Displacement studies were performed at expected trough levels and were performed at three dose levels (0.42, 1.25, and 3.75 mg / kg). The 3.75 mg / kg dose was able to achieve >90% receptor occupancy at trough.

[0228] PET imaging and immunohistochemistry study in an APP / PS1 mouse model with a drug-withdrawn period The objective of this study was to assess synaptic density in control and APP / PS1 mice before and after treatment with Compound 1, as well as to assess the reversibility of the observed effects after a drug withdrawal period.

[0229] Vehicle (95% polyethylene glycol (PEG) 400 / 5% solutol) or Compound 1 was administered orally at 7.5 mg / kg / day (3.75 mg / kg q12) for at least 28 days to control and APP / PS1 mice. A portion of the mice discontinued treatment after 4 weeks, followed by a 4-week washout period.

[0230] To further maximize translational relevance, other studies aged mice to a point where synapse loss was detectable using SV2A PET. At 12 months of age, hippocampal / brainstem [ 18 Synaptic density, as measured by [F]SDM-8 SUVR-1, is reduced in APP / PS1 mice compared to wild-type. Importantly, re-examination of the same mice after a one-month course of treatment with compound 1 showed that synaptic density had increased significantly to levels consistent with wild-type mice.

[0231] The decrease in synaptic density in APP / PS1 samples compared to controls was confirmed by immunohistochemical staining of pre- and postsynaptic markers (SV2A and PSD-95) in hippocampal and cortical sections. Significant increases in both markers and in both regions were observed in APP / PS1 samples treated with Compound 1 (compared to vehicle-treated samples). The observed recovery in synaptic density was maintained after a one-month washout period.

[0232] SV2A PET imaging and APP by immunohistochemistry NLGF Testing the / MAPT Mouse Model The purpose of this study was to determine the effect of Compound 1 on the expression of control and NL-GF mutant amyloid precursor protein / microtubule-associated protein tau (APP) before and after treatment. NLGFThe aim of this study was to evaluate synaptic density in control and dKI (dKI-double knock-in) mice. Vehicle (95% PEG-400 / 5% Solutol) or Compound 1 was administered orally at 7.5 mg / kg / day (3.75 mg / kg q12) to control and dKI mice for at least 28 days.

[0233] To further maximize translational relevance, other studies aged mice to a point where synapse loss was detectable using SV2A PET. At 12 months of age, hippocampal / brainstem [ 18 Synaptic density, as measured by [F]SDM-8 SUVR-1, was reduced in dKI mice compared with wild-type mice. Importantly, re-examination of the same mice after a 1-month course of treatment with compound 1 showed a highly significant increase in synaptic density.

[0234] Immunohistochemical staining of pre- and postsynaptic markers (SV2A and PSD-95) in hippocampal and cortical sections confirmed a decrease in synaptic density in dKI samples versus controls. Compound 1-treated dKI samples showed a significant increase in SV2A in the hippocampus and both markers in the cortex (compared to vehicle-treated samples).

[0235] Prevention of Positive Allosteric Regulator-Induced Seizures The purpose of this study was to evaluate the enantiomeric selectivity of compound 1 for the mGluR5 receptor as a function of seizure prevention. PAMs of mGluR5 have previously been reported to induce seizures in C57 / Bl6J mice. Also, a single 7.5 mg / kg dose of compound 1 has previously been shown to prevent seizures induced by (4R,5R)-5-(4-fluorophenyl)-4-(5-((5-fluoropyridin-3-yl)ethynyl)pyridin-3-yl)oxazolidin-2-one (PAM).

[0236] Compound 1 (0.12, 0.24, 0.47, 0.94, 1.88, 3.75 or 7.5 mg / kg), enantiomers of Compound 1 (7.5, 15 or 30 mg / kg), or vehicle were administered by oral gavage to 3-11 month old C57 / Bl6J mice. Two hours later, PAM compound was administered by intraperitoneal injection at 20 mg / kg. Animals were then placed in 10 inch diameter acrylic cages for observation and video recording for 2 hours post-administration, and seizures were scored using the Racine seizure scoring criteria and standardized to a 0-1 scale.

[0237] Compound 1 dose-dependently prevented mGluR5 PAM-induced seizure activity in C57 / Bl6J mice. Compound 1 (SAM in Figure 2) inhibited PAM-induced seizures with an IC50 of 1.09 mg / kg (Figure 2). Pretreatment with a high dose of the enantiomer of compound 1 (eSAM in Figure 2) did not prevent seizure activity.

[0238] The anticonvulsant effect of compound 1 demonstrated dose-dependent occupancy of mGluR5 receptors in the brains of C57 / Bl6J mice. High doses of the enantiomers of compound 1 did not reduce convulsive activity, suggesting that receptor binding is stereoselective.

[0239] Example 6. Phase I Clinical Trial Study Design Overview This was an open-label, single ascending dose (SAD) study to evaluate the safety of a single oral dose of Compound 1 in healthy men and women aged 50 to 80 years (inclusive) with no history of cognitive impairment.

[0240] Six cohorts, each with six patients, were assigned to receive a single oral dose of Compound 1 in the fasted state at the following dose levels: 10, 40, 70, 100, 150, and 200 mg, followed by a 7-day follow-up period. The starting dose of 10 mg was <10% of the rat NOAEL human equivalent dose (HED) of 140 mg in a GLP-compliant 28-day 15 mg / kg rat study.

[0241] The study medication was a capsule containing 5 mg, 50 mg, or 100 mg of nanomilled active pharmaceutical ingredient, and each cohort's dose was achieved by using a combination of these capsules. Doses were administered in an inpatient setting, and all participants were closely monitored before discharge on day 3, with follow-up visits on days 4 and 7, and a phone call on day 5 to inquire about their general health.

[0242] Dosing of Compound 1 was carefully monitored within and between cohorts to ensure patient safety, and the decision to open the next dose cohort was made after all participants in a given cohort had completed the study and all available clinical and safety data had been reviewed by the principal investigator, medical monitor, DSMB, and IND sponsor.

[0243] After study drug administration, vital signs were monitored hourly for the first 8 hours and every 3 hours until discharge on day 3, and ECGs were performed approximately 6, 24, and 48 hours after dosing. To monitor changes in consciousness, Glasgow Coma Scale (GCS) was performed before dosing and every 2 hours for the first 8 hours, then every 3 hours until discharge on day 3. To monitor cognitive or psychiatric side effects, MOCA, GDS, and NPIQ were performed before oral dosing, approximately 6 hours after dosing (estimated peak concentration), and approximately 24 hours after dosing. Adverse events were monitored continuously. Blood was collected for safety laboratory tests before drug dose administration and on days 2 and 3.

[0244] Participant disposition, demographics and baseline characteristics A total of 36 patients (6 in each dose cohort) enrolled and completed the study. Across the cohort, mean age and body mass index (BMI) at baseline ranged from 68.3 to 72.8 years and 24.8 to 27.8 years, respectively.

[0245] The majority of participants were white (97%) and female (58.3%), and mean scores on cognition and depression measures administered at baseline did not indicate signs of cognitive impairment or clinically significant depression (Table 20).

[0246] Table 20. Participant demographics and baseline characteristics [Table 22] Abbreviations: BMI = body mass index, CDR = Clinical Dementia Rating Scale, GCS = Glasgow Coma Scale, GDS = Geriatric Depression Rating Scale, MMSE = Mini-Mental State Examination, MoCA = Montreal Cognitive Assessment, SD = standard deviation. 1 A score above 0 indicates progressive decline due to cognitive impairment. 2 Scores range from 1 to 19. Higher scores indicate better delayed speech recall. 3 Scores range from 1 to 30. Higher scores indicate better cognitive function. 4 Scores range from 3 to 15. Lower scores indicate greater impairment of consciousness. 5 Scores range from 0 to 15. Higher scores indicate a higher likelihood of depression. 6 Scores range from 0 to 30. Higher scores indicate better cognitive function.

[0247] safety results A single oral dose of compound 1 was well tolerated with no deaths, serious SAEs, or serious TEAEs observed in all participants. There was also no evidence of clinically significant changes in consciousness as measured by GCS, or cognitive or psychiatric side effects as measured by MOCA, GDS, or NPIQ in all participants. All TEAEs were mild or moderate in severity, and eight TEAEs were considered possibly related to treatment; all others were rated as unlikely related. These TEAEs consisted of three brief cases of mouth sensation (abnormal taste, tingling tongue, and mouth pain), one brief case of dizziness, two cases of transient headache (one of which was treated with a single dose of acetaminophen 500 mg), one case of transient hypertension, and one case of elevated triglyceride laboratory values ​​on day 7 in a participant with a history of hypertriglyceridemia.

[0248] Equivalence and Scope Those skilled in the art will be able to recognize or ascertain, given the disclosure set forth herein, without using undue routine experimentation, many equivalents to the specific embodiments. The scope of the present disclosure is not intended to be limited to the above description, but rather is as defined in the appended claims.

[0249] In the claims, articles such as "a," "an," and "the" may mean one or more, unless indicated to the contrary or otherwise clear from the context. A claim or description containing "or" between one or more elements of a group is deemed to be satisfied if one, more than one, or all of the elements of the group are present in, used in, or otherwise relevant to a given product or process, unless indicated to the contrary or otherwise clear from the context. The disclosure includes embodiments in which exactly one of the elements of the group is present in, used in, or otherwise relevant to a given product or process. The disclosure includes embodiments in which more than one of the elements of the group, or all of the elements of the group are present in, used in, or otherwise relevant to a given product or process.

[0250] Additionally, the term "comprising" is intended to be inclusive and permits, but does not require, the inclusion of additional elements or steps. When the term "comprising" is used herein, the term "consisting of" is also included and disclosed.

[0251] When ranges are given, the endpoints are inclusive. Furthermore, unless otherwise indicated or otherwise clear from the context and the understanding of one of ordinary skill in the art, values ​​given as ranges should be understood to be able to assume, in different embodiments of this disclosure, any specific value or subrange within the given range, to one-tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0252] Furthermore, it should be understood that if a particular embodiment of the present disclosure falls within the prior art, that embodiment may be expressly excluded from one or more of the claims. Such embodiments may be excluded even if the exclusion is not expressly set forth herein, since they are deemed to be well known to those of skill in the art. Particular embodiments of the compositions of the present disclosure (e.g., any antibiotic, therapeutic agent, or active ingredient, any method of manufacture, any method of use, etc.) may be excluded from one or more of the claims for any reason, whether related to the existence of prior art or not.

[0253] The words which have been used are to be understood as words of description rather than of limitation, and changes may be made within the purview of the appended claims without departing from the true scope and spirit of the disclosure in its broader aspects.

[0254] While this disclosure has been described at some length and with some specificity with respect to certain depicted embodiments, it is not intended that the disclosure be limited to such particulars or embodiments, or to any particular embodiment, but rather, it is necessary to interpret the appended claims in order to accord such claims the broadest possible interpretation in view of the prior art, so as to effectively encompass the intended scope of the disclosure.

Claims

1. When measured by X-ray diffraction using Cu Kα X-ray irradiation, the anhydrous (AH) crystalline form (Morphology A) of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridine-3-yl)oxazolidine-2-one (compound 1) has an XRPD pattern containing peaks at 2θ angles of 9.02, 11.65, and 11.86 degrees 2θ, each ±0.2 degrees 2θ. 【Chemistry 1】

2. The AH crystal form according to claim 1, wherein, when measured by X-ray diffraction measurement induced by Cu Kα X-ray irradiation, the XRPD pattern further includes peaks at 2θ angles of 12.15, 14.99, 28.99, and 44.03 degrees 2θ, each by ±0.2 degrees 2θ.

3. The AH crystal form according to claim 1, wherein, when measured by X-ray diffraction measurement induced by Cu Kα X-ray irradiation, the XRPD pattern further includes three peaks at 21.09, 21.49, and 21.88 degrees 2θ, and each at ±0.2 degrees 2θ.

4. The AH crystal form according to claim 1, wherein, when measured by X-ray diffraction measurement induced by Cu Kα X-ray irradiation, the XRPD pattern includes peaks at 9.02, 11.65, 11.86, 12.15, 14.99, 21.09, 21.49, 21.88, 28.99, and 44.03 degrees 2θ.

5. The AH crystal form according to claim 1, having substantially the XRPD pattern shown in Figure 3.

6. The AH crystal form according to claim 1, having a melting endothermic peak at approximately 134°C to 138°C in a differential scanning calorimetry (DSC) thermogram.

7. The AH crystal form according to claim 1, having a DSC thermogram which is substantially the DSC graph shown in Figure 4.

8. The AH crystal form according to claim 1, wherein the weight loss by thermogravimetric analysis (TGA) is approximately 0.02% (w / w) between ambient temperature and approximately 250°C.

9. The AH crystal form according to claim 1, having a TGA which is substantially the TGA graph shown in Figure 4.

10. A method for preparing the anhydrous (AH) crystalline form of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridine-3-yl)oxazolidine-2-one (compound 1), comprising precipitating the anhydrous crystalline form from a solution containing compound 1 and a solvent selected from the group consisting of ethyl acetate (Â), heptane, and mixtures thereof.

11. When measured by X-ray diffraction by Cu Kα X-ray irradiation, the solvate form (Form B) of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridine-3-yl)oxazolidine-2-one (Compound 1) has an XRPD pattern containing at least three peaks at the following 2θ angles: 16.07, 16.27, 16.58, and 16.77 degrees 2θ, each ±0.2 degrees 2θ.

12. A method for preparing solvate form B of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridine-3-yl)oxazolidine-2-one (compound 1), comprising evaporation of a mixture of compound 1 and isopropyl alcohol in an inert atmosphere in a drying box.

13. When measured by X-ray diffraction using Cu Kα X-ray irradiation, the solvate form (Form C) of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridine-3-yl)oxazolidine-2-one (Compound 1) has an XRPD pattern containing only two peaks between 13.32 and 13.82 degrees 2θ, each at ±0.2 degrees 2θ.

14. A method for preparing the solvate form C of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridine-3-yl)oxazolidine-2-one (compound 1), comprising a precipitate of the solvate form from a solution containing compound 1 and isopropyl alcohol.

15. A method for preparing the solvate form C of (4R,5R)-5-(2-chlorophenyl)-4-(5-(phenylethynyl)pyridine-3-yl)oxazolidine-2-one (compound 1), (i) A step of dissolving compound 1 in isopropyl alcohol to form a saturated solution, (ii) Filtering the mixture from step (i) into a new vial placed in the outer vial containing the reverse solvent, (iii) a step of stirring the mixture in the vial inside step (ii) to precipitate the solid, and (iv) A step of filtering the solid from step (iii) and vacuum drying the solid at room temperature. A method that includes this.

16. A pharmaceutical composition comprising an effective amount of the AH crystalline form described in claim 1.

17. The pharmaceutical composition according to claim 16, comprising about 5% (w / w) to about 30% (w / w) of AH crystalline form.

18. The pharmaceutical composition according to claim 16, further comprising a pharmaceutically acceptable plasticizer, binder, filler, carrier, excipient, lubricant, disintegrant, and / or surfactant.

19. The pharmaceutical composition according to claim 16, comprising the components listed in the following table. However, the amounts of binders, surfactants, carriers, disintegrants, and lubricants are not all 0%.

20. The pharmaceutical composition according to claim 19, wherein the binder is hypromellose, the surfactant is sodium lauryl sulfate, the carrier and binder is lactose monohydrate, the disintegrant is croscarmellose, and the lubricant is magnesium stearate.

21. A pharmaceutical composition comprising a therapeutically effective amount of the crystalline or solvated form described in claim 1 for the treatment of Alzheimer's disease, prevention of seizures, restoration of synaptic loss, or reduction of tau accumulation in a subject requiring treatment.

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