Crystalline forms of trifinetide

Abstract

The present application relates to crystalline forms of trifinetide. The present disclosure provides: crystalline forms of trafennetide and trafennetide hydrate; a pharmaceutical composition comprising a crystalline form of trafenotide and a trafenotide hydrate; a process for the preparation of a crystalline form of trifinetide or trifinetide hydrate; and a method of treating a disease, disorder, or condition in a subject comprising administering to the subject a composition comprising a crystalline form of trifinetide or trifinetide hydrate.

Description

[0001] This application is a divisional application of the application filed on July 12, 2022, with application number 202280041351.X and invention title "Crystal Form of Trifenexy". Technical Field

[0002] This disclosure provides: crystalline forms of trafeneptide and trafeneptide hydrate; pharmaceutical compositions comprising crystalline forms of trafeneptide and trafeneptide hydrate; methods for preparing crystalline forms of trafeneptide or trafeneptide hydrate; and methods for treating a subject with a disease, condition, or symptom, comprising administering to the subject a composition comprising crystalline forms of trafeneptide or trafeneptide hydrate. Background Technology

[0003] Glycyl-L-2-methylprolyl-L-glutamate (also known as trifineptide) is a synthetic analogue of glycine-proline-glutamate (also known as glypromate or GPE). GPE is naturally found in the brain. It is the N-terminal tripeptide of the insulin-like growth factor 1 (IGF-1) protein.

[0004] US 7,041,314 discloses trafenexide, methods for preparing trafenexide, and methods for using trafenexide to treat diseases, conditions, or symptoms (e.g., neurodegeneration caused by hypoxic-ischemic or toxic injury). US 7,605,177 discloses methods for using trafenexide to treat diseases such as neurodegenerative and chronic neurodegenerative conditions, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, diabetic neuropathy caused by type I or type II diabetes, autoimmune brain diseases, or multiple sclerosis. US 7,714,020 discloses methods for using trafenexide to treat diseases, conditions, or symptoms such as brain injury caused by traumatic brain injury, stroke, hypoxic / ischemic, and toxic injury. US 7,863,304 discloses methods for using trafenexide to treat diseases, conditions, or symptoms such as chronic neurodegenerative conditions, such as Parkinson's disease. US 8,637,567 discloses a method of treating a disease, condition, or symptom using trafeneptide, such as cognitive impairment or memory impairment. US 7,887,839, US 8,178,125, and US 8,496,963 disclose oral formulations of trafeneptide for treating a variety of diseases, conditions, or symptoms. US 9,708,366 and US 9,212,204 disclose methods of treating a disease, condition, or symptom using trafeneptide, such as autism spectrum disorders, such as autism, autism disorder Asperger's syndrome, childhood disintegration disorder, pervasive developmental disorder (PDD-NOS) not otherwise specified, fragile X syndrome, or Rett syndrome. Summary of the Invention

[0005] The chemically stable trofenexyte solid form is required for use in treating subjects with Rett syndrome, Fragile X syndrome, traumatic brain injury, and other diseases, conditions, and symptoms.

[0006] On the one hand, this disclosure provides a crystalline polymorph of trefeneptide or trefeneptide hydrate.

[0007] On the other hand, this disclosure provides a method for preparing the crystalline polymorphic form of trafenesin.

[0008] On the other hand, this disclosure provides a composition comprising a crystalline polymorph of trafenesin or trafenesin hydrate and one or more excipients.

[0009] On the other hand, this disclosure provides a method for preparing a composition comprising a crystalline polymorph of trafenesin or trafenesin hydrate and one or more excipients.

[0010] On the other hand, this disclosure provides a method for treating a subject’s disease, condition or symptom using trafenexide or a crystalline polymorph of trafenexide hydrate, such as traumatic brain injury or neurodevelopmental disorder.

[0011] On the other hand, this disclosure provides a kit comprising a crystalline polymorph of trafenestrate or trafenestrate hydrate. Attached Figure Description

[0012] Figure 1 The XRPD diffraction pattern is for form A.

[0013] Figure 2 The Raman spectrum is of form A.

[0014] Figure 3 The LF-Raman spectrum is for form A.

[0015] Figure 4 The ssNMR spectrum is of form A.

[0016] Figure 5 This is the DSC thermogram of form A.

[0017] Figure 6 Here is the IR spectrum of form A.

[0018] Figure 7 Here is the NIR spectrum of form A.

[0019] Figure 8 This represents the single-crystal X-ray diffraction asymmetric unit of form A. For clarity, hydrogen atoms have been omitted.

[0020] Figure 9 A line graph showing the dynamic vapor adsorption / desorption data for Form A.

[0021] Figure 10 Three XRPD diffraction pattern series for form A containing different amounts of water.

[0022] Figure 11 Five XRPD diffraction pattern series for form A stored under different humidity conditions.

[0023] Figure 12 The TGA / DCS thermogram is for form A. Detailed Implementation

[0024] I. Crystalline trefeneptide or trefeneptide hydrate In one embodiment, this disclosure provides a crystalline polymorph of triphenide or a crystalline polymorph of triphenide hydrate, collectively referred to as "triphenide polymorph".

[0025] In another embodiment, the trafenexylidene polymorph is a crystalline trafenexylidene hydrate represented by the formula: trafenexylidene·xH2O, where x is from about 2 to about 4. In another embodiment, the trafenexylidene polymorph is trafenexylidene·xH2O, where x is from about 2.5 to about 3.5. In another embodiment, the trafenexylidene polymorph is trafenexylidene·xH2O, where x is about 2. In another embodiment, the trafenexylidene polymorph is trafenexylidene·xH2O, where x is about 2.5. In another embodiment, the trafenexylidene polymorph is trafenexylidene·xH2O, where x is about 3. In another embodiment, the trafenexylidene polymorph is trafenexylidene·xH2O, where x is about 3.5. In another embodiment, the trafenexylidene polymorph is trafenexylidene·xH2O, where x is about 4. These crystalline trefeneptide xH2O polymorphs are collectively referred to as "Form A".

[0026] In another embodiment, form A is characterized by having a PXRD pattern with peaks in the range of 6.6–6.8 degrees 2Θ, 11.3–11.6 degrees 2Θ, 12.5–12.7 degrees 2Θ, and 13.6–13.8 degrees 2Θ (where the 2Θ value is ±0.2 degrees 2Θ) when irradiated with Cu Kα, and optionally, peaks at 16.8, 22.3, 23.6, 25.3, and / or 28.1 degrees 2Θ ±0.2 degrees 2Θ when irradiated with Cu Kα.

[0027] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, with peaks at 6.7 or 6.8, 11.4 or 11.5, 12.6 and 13.7 or 13.8 degrees 2Θ and optionally, peaks at 16.8, 22.3, 23.6, 25.3 and / or 28.1 degrees 2Θ ± 0.2 degrees 2Θ.

[0028] In another embodiment, form A is characterized by having a PXRD pattern with peaks at 6.7, 11.4, 12.6, 13.7, 22.3, 23.6, 25.3 and 28.1 degrees 2Θ using Cu Kα radiation, where the 2Θ value is ±0.2 degrees 2Θ.

[0029] In another embodiment, form A is characterized by having a PXRD pattern with peaks at 2Θ degrees 6.7, 11.4, 12.6, 13.7, 22.3, 23.6, 25.3 and 28.1 degrees using Cu Kα radiation.

[0030] In another embodiment, form A is characterized by having a PXRD pattern with peaks at 6.8, 11.5, 12.6, 13.8 and 16.8 degrees 2Θ using Cu Kα radiation, where the 2Θ value is ±0.2 degrees 2Θ.

[0031] In another embodiment, form A is characterized by having a PXRD pattern with peaks at 11.5, 12.6, and 13.8 degrees 2Θ using Cu Kα radiation, where the 2Θ value is ±0.2 degrees 2Θ.

[0032] In another embodiment, form A is characterized by having a PXRD pattern with peaks at 6.8, 11.5, and 12.6 degrees 2Θ using Cu Kα radiation, where the 2Θ value is ±0.2 degrees 2Θ.

[0033] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least three peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0034] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least four peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0035] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least five peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0036] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least six peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0037] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least seven peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0038] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least eight peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0039] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least nine peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0040] In another embodiment, form A is characterized by having a PXRD pattern using Cu Kα radiation, having at least ten peaks at 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.7, 16.3, 16.8, 18.2, 18.7, 19.0, 19.2, 19.8, 20.4, 20.8, 21.2, 21.5, 22.3, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4 and / or 39.1 degrees 2Θ, where the 2Θ value is ±0.2 degrees 2Θ.

[0041] In another embodiment, form A is characterized by having a PXRD pattern with peaks at 2Θ of 6.8, 9.9, 11.5, 12.6, 13.5, 13.8, 13.9, 15.4, 15.8, 16.3, 16.8, 18.2, 18.7, 19.0, 19.270, 19.8, 20.4, 20.8, 21.2, 21.5, 22.380, 22.8, 23.3, 23.6, 25.4, 26.4, 28.1, 30.2, 32.9, 33.7, 35.4, and 39.120 degrees, where the 2Θ value is ±0.2 degrees 2Θ.

[0042] In another embodiment, form A is characterized by having PXRD plots of d-spacing at 13.1, 7.7, 7.0, 6.4 and 5.3 Å using Cu Kα radiation.

[0043] In another embodiment, form A is characterized by having PXRD plots with d-spacing at 13.1, 7.7, and 7.0 Å using Cu Kα radiation.

[0044] In another embodiment, form A is characterized by having PXRD plots with d-spacing at 7.7, 7.0, and 6.4 Å using Cu Kα radiation.

[0045] In another embodiment, form A is characterized by having PXRD patterns with at least three d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5, and / or 2.3 Å, using Cu Kα radiation.

[0046] In another embodiment, form A is characterized by having PXRD patterns with at least four d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5, and / or 2.3 Å, using Cu Kα radiation.

[0047] In another embodiment, form A is characterized by having PXRD patterns with at least five d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5 and / or 2.3 Å, using Cu Kα radiation.

[0048] In another embodiment, form A is characterized by having PXRD patterns with at least six d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5, and / or 2.3 Å, using Cu Kα radiation.

[0049] In another embodiment, form A is characterized by having PXRD patterns with at least seven d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5, and / or 2.3 Å, using Cu Kα radiation.

[0050] In another embodiment, form A is characterized by having PXRD patterns with at least eight d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5, and / or 2.3 Å, using Cu Kα radiation.

[0051] In another embodiment, form A is characterized by having PXRD patterns with at least nine d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5, and / or 2.3 Å, using Cu Kα radiation.

[0052] In another embodiment, form A is characterized by having PXRD patterns with at least ten d-spacings at 13.1, 8.9, 7.7, 7.0, 6.6, 6.43, 6.36, 5.8, 5.6, 5.4, 5.3, 4.9, 4.8, 4.7, 4.6, 4.5, 4.34, 4.27, 4.2, 4.1, 4.0, 3.9, 3.81, 3.76, 3.5, 3.4, 3.2, 3.0, 2.72, 2.66, 2.5 and / or 2.3 Å, using Cu Kα radiation.

[0053] In another implementation, form A is characterized by having the same Figure 1 The PXRD diffraction pattern depicted in the image is essentially the same as the PXRD diffraction pattern in the image.

[0054] In another embodiment, form A is characterized by having 2989, 2934, 2883, 1685, 1637, 1459 and 930 cm -1 The FT-Raman spectrum with peaks at cm -1 The value is ±4 cm -1 .

[0055] In another embodiment, form A is characterized by having 2989, 2960, 2934, 2883, 1685, 1637, 1459, 1417, 1346, 1272, 1199, 1058, 1023, 967, 930, 782, 552, 496, 425 and 342 cm -1 The FT-Raman spectrum with peaks at cm -1 The value is ±4 cm -1 .

[0056] In another implementation, form A is characterized by having the same Figure 2 The FT-Raman spectra depicted in the figure are essentially the same as the FT-Raman spectra.

[0057] In another embodiment, form A is characterized by having 13, 24, 67, and 77 cm. -1 The low-frequency (LF) Raman spectrum with a peak at cm -1 The value is ±4 cm -1 .

[0058] In another embodiment, form A is characterized by having 13, 24, 34, 67, 77, 208, 283, 348, 422, 495, and 552 cm. -1 The LF-Raman spectrum with peaks at cm -1 The value is ±4 cm -1 .

[0059] In another implementation, form A is characterized by having the same Figure 3 The LF-Raman spectra depicted are essentially the same as the LF-Raman spectra.

[0060] In another embodiment, form A is characterized by having peaks at 179.7, 177.9, 177.5, 177.2, 177.0, 165.3, 164.9, 164.8, 67.8, 67.4, 58.6, 58.2, 46.8, 40.3, 33.3, 25.3, 23.5, and 21.1 ppm. 13 Solid-state nuclear magnetic resonance (ssNMR) spectrum of C, with ppm values ​​of ±3 ppm.

[0061] In another embodiment, form A is characterized by having an ssNMR spectrum with 18 peaks, wherein: (i) the Δ from the lowest field peak to the second lowest field peak is 1.8 ppm; (ii) the Δ from the lowest field peak to the third lowest field peak is 2.2 ppm; (iii) the Δ from the lowest field peak to the fourth lowest field peak is 2.5 ppm; (iv) the Δ from the lowest field peak to the fifth lowest field peak is 2.7 ppm; (v) the Δ from the lowest field peak to the sixth lowest field peak is 14.4 ppm; (vi) the Δ from the lowest field peak to the seventh lowest field peak is 14.8 ppm; (vii) the Δ from the lowest field peak to the eighth lowest field peak is 14.9 ppm; (viii) the Δ from the lowest field peak to the ninth lowest field peak is 111.9 ppm; (ix) the Δ from the lowest field peak to the tenth lowest field peak is 112.3 ppm; and (x) the Δ from the lowest field peak to the eleventh lowest field peak is 121.1 ppm. ppm; (xi) Δ from the lowest field peak to the twelfth lowest field peak is 121.5 ppm; (xii) Δ from the lowest field peak to the thirteenth lowest field peak is 133.1 ppm; (xiii) Δ from the lowest field peak to the fourteenth lowest field peak is 139.4 ppm; (xiv) Δ from the lowest field peak to the fifteenth lowest field peak is 146.3 ppm; (xv) Δ from the lowest field peak to the sixteenth lowest field peak is 154.4 ppm; (xvi) Δ from the lowest field peak to the seventeenth lowest field peak is 156.2 ppm; and / or (xvii) Δ from the lowest field peak to the highest field peak is 158.6 ppm, or any combination thereof. See Tables 5 and 6 for example.

[0062] In another implementation, form A is characterized by having the same Figure 4 The ssNMR spectrum depicted in the image is essentially the same as the ssNMR spectrum.

[0063] In another embodiment, form A is characterized by having a melting point based on differential scanning calorimetry (DSC), with an initial temperature of 71.71°C and a peak temperature of 72.06°C.

[0064] In another implementation, form A is characterized by having the same Figure 5 The DSC thermograms depicted are essentially the same as those in the original DSC thermograms.

[0065] In another embodiment, form A is characterized by having 1678, 1636, 1589, 1525, 1214 and 1196 cm. -1 The infrared (IR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 .

[0066] In another embodiment, form A is characterized by having 3560, 3400, 3343, 3296, 2881-3012, 1678, 1636, 1589, 1525, 1458, 1435, 1413, 1376, 1352, 1292, 1255, 1214, 1196, 1142, 1120, 1015, 964, 924, 898, 827, 843, 777, 649, 599, 576, 551, 502 and 426 cm -1 The IR spectrum with a peak at cm -1 The value is ±4 cm -1 .

[0067] In another implementation, form A is characterized by having the same Figure 6 The IR spectra depicted in the image are essentially the same as the IR spectra.

[0068] In another embodiment, form A is characterized by having 5145, 4630, and 4423 cm. -1 The near-infrared (NIR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 .

[0069] In another embodiment, form A is characterized by having 5908, 5796, 5145, 4875, 4630, 4423 and 4298 cm. -1 NIR spectrum with peak at cm -1 The value is ±4 cm -1 .

[0070] In another implementation, form A is characterized by having the same Figure 7 The NIR spectra depicted in the text are essentially the same as the NIR spectra.

[0071] In another embodiment, the trafeneptide polymorph (e.g., form A) is characterized by containing about 1% to about 10%, for example about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, or about 1% by weight of any other physical form, such as crystalline or amorphous form, of trafeneptide or trafeneptide hydrate.

[0072] In another embodiment, the trafeneptide polymorph (e.g., form A) is characterized by comprising about 0.1% to about 1%, for example about 1%, about 0.9%, about 0.8%, about 0.7%, about 0.6%, about 0.5%, about 0.4%, about 0.3%, about 0.2% or about 0.1% by weight of any other physical form of trafeneptide or trafeneptide hydrate.

[0073] In another embodiment, the trafenexide polymorph is characterized by not containing any other physical form of trafenexide or trafenexide hydrate in a PXRD-detectable amount. In another embodiment, the trafenexide polymorph is form A.

[0074] In another embodiment, the average particle size distribution of the trafenexide polymorph is from about 10 µm to about 500 µm, for example, about 500 µm, about 400 µm, about 300 µm, about 200 µm, about 100 µm, about 90 µm, about 80 µm, about 70 µm, about 60 µm, about 50 µm, about 40 µm, about 30 µm, about 20 µm, or about 10 µm. In another embodiment, the trafenexide polymorph is form A.

[0075] In another embodiment, the average particle size distribution of the trafenexyte polymorph is from about 1 µm to about 10 µm, for example, about 10 µm, about 9 µm, about 8 µm, about 7 µm, about 6 µm, or about 5 µm, about 4 µm, about 3 µm, about 2 µm, or about 1 µm. In another embodiment, the trafenexyte polymorph is form A.

[0076] In another embodiment, the average particle size distribution of the trafenexylidene polymorph is about 1 µm or less, for example, about 0.9 µm, about 0.8 µm, about 0.7 µm, about 0.6 µm, about 0.5 µm, about 0.4 µm, about 0.3 µm, about 0.2 µm, about 0.1 µm, about 0.09 µm, about 0.08 µm, about 0.07 µm, about 0.06 µm, about 0.05 µm, about 0.04 µm, about 0.03 µm, about 0.02 µm, or about 0.01 µm or less. In another embodiment, the trafenexylidene polymorph is form A.

[0077] In another embodiment, the trifenexylidene polymorph is chemically stable after storage for 3 months at a temperature of about 25°C and a relative humidity of about 60%. In another embodiment, the trifenexylidene polymorph is in form A.

[0078] In another embodiment, the trifenexylidene polymorph is chemically stable after storage for 6 months at a temperature of about 25°C and a relative humidity of about 60%. In another embodiment, the trifenexylidene polymorph is in form A.

[0079] In another embodiment, the trifenexylidene polymorph is chemically stable when stored at a temperature of about 25°C and a relative humidity of about 60% for 12 months or longer. In another embodiment, the trifenexylidene polymorph is in form A.

[0080] II. Pharmaceutical compositions and formulations In another embodiment, this disclosure provides a pharmaceutical composition comprising a trafenexylidene polymorph and one or more pharmaceutically acceptable excipients.

[0081] In another embodiment, this disclosure provides a pharmaceutical formulation comprising a trafenetide polymorph in particulate form, wherein the particulates optionally comprise one or more pharmaceutically acceptable binders or fillers, or combinations thereof. The binder may be used in a total amount of about 1% to about 30% of the particulates by weight, for example, in a total amount of about 5% to about 15% of the particulates by weight, for example, in a total amount of about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% of the particulates by weight. The filler may be used in a total amount of about 5% to about 80% of the particles by weight, for example, in a total amount of about 10% to about 60% of the particles by weight, for example, in a total amount of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75% or about 80% of the particles by weight.

[0082] In another embodiment, the adhesive is gum arabic, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, polyethylene glycol (PEG), povidone (polyvinylpyrrolidone, PVP), sucrose, or starch, or a combination thereof.

[0083] In another embodiment, the filler is microcrystalline cellulose.

[0084] In another embodiment, this disclosure provides an aqueous pharmaceutical formulation comprising a trifenestrated peptide polymorph dissolved in water and optionally one or more other excipients.

[0085] In another implementation, the water is purified water.

[0086] In another embodiment, approximately 1 gram of trefenapyline polymorph is dissolved in every 5 mL of water.

[0087] In another embodiment, the trefeneptide polymorph is form A.

[0088] In another embodiment, the pharmaceutical formulation contains 1% to 99% by weight of the trifenetide polymorph, such as about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. The amount in any particular formulation will depend on the effective dose of trifenetide, i.e., the dose required to induce the desired level of therapeutic activity. In one embodiment, the pharmaceutical formulation comprises form A dissolved in water.

[0089] III. Methods for preparing pharmaceutical formulations In another embodiment, this disclosure provides a method for preparing an aqueous pharmaceutical formulation comprising trafeneptide, the method comprising dissolving a polymorph of trafeneptide in water.

[0090] In another implementation, the water is purified water.

[0091] In another embodiment, approximately 1 gram of trefeneptide polymorph is dissolved in 5 mL of water.

[0092] In another embodiment, the trefeneptide polymorph is form A.

[0093] IV. Medicine Box In another embodiment, this disclosure provides a medicine box comprising a tripfenide polymorph, said tripfenide polymorph being packaged in a manner that facilitates its use in practicing the methods of this disclosure.

[0094] In another embodiment, the kit includes a trifenexylidene polymorph packaged in a container (such as a sealed bottle or dish), with a label affixed to the container or an insert included in the kit describing the use of trifenexylidene in practicing the methods of this disclosure for the treatment of a subject's disease, condition, or symptom. In another embodiment, the trifenexylidene polymorph is packaged in unit dosage form.

[0095] In another embodiment, the kit also includes instructions for dissolving the trifenestrated peptide polymorph in water to provide an aqueous pharmaceutical formulation.

[0096] In another embodiment, the kit may also include inserts, such as instructions for administering the trifenestrate polymorph or aqueous pharmaceutical preparation to a subject suffering from a disease, condition, or symptom. In another embodiment, the disease, condition, or symptom is traumatic brain injury. In another embodiment, the disease, condition, or symptom is a neurodevelopmental disorder. In another embodiment, the neurodevelopmental disorder is Rett syndrome, Fragile X syndrome, or autism spectrum disorder.

[0097] In another embodiment, the trefeneptide polymorph is form A.

[0098] V. Methods for treating diseases, symptoms, or illnesses In another embodiment, this disclosure provides a method for treating a disease, condition, or symptom in a subject of need, the method comprising administering to the subject an aqueous pharmaceutical preparation comprising a trifenetide polymorph dissolved in water. In another embodiment, the disease, condition, or symptom is traumatic brain injury. In another embodiment, the disease, condition, or symptom is a neurodevelopmental disorder. In another embodiment, the neurodevelopmental disorder is Rett syndrome, Fragile X syndrome, or autism spectrum disorder.

[0099] In another embodiment, the aqueous pharmaceutical preparation is a solution for oral administration.

[0100] In another implementation, the water is purified water.

[0101] In another embodiment, approximately 1 gram of trefeneptide polymorph is dissolved in 5 mL of water.

[0102] In another embodiment, the trefeneptide polymorph is form A.

[0103] VI. Method for preparing crystalline trefeneptide or trefeneptide hydrate In another embodiment, this disclosure provides a method for preparing trefeneptide polymorphs.

[0104] In another embodiment, this disclosure provides a method for preparing form A.

[0105] In another embodiment, this disclosure provides a method for preparing form A, the method comprising i) adding ethanol to an aqueous solution of trafenexylidene at about 25°C; and ii) cooling the solution to about 0°C. In another embodiment, the water:ethanol ratio is about 3:7 w / w. In another embodiment, the solution concentration of trafenexylidene is about 15% w / w.

[0106] In another embodiment, the method for preparing form A further includes adding form A to seed crystals in the solution to obtain a slurry.

[0107] In another embodiment, the method for preparing form A further includes, for example, separating the obtained solid by filtration to obtain a wet filter cake containing form A.

[0108] In another embodiment, the method for preparing form A further includes washing the wet filter cake containing form A with pre-cooled ethanol at about 0°C.

[0109] In another embodiment, the method for preparing form A further includes drying the wet filter cake containing form A under vacuum.

[0110] VII. Definition As used herein, the term "trifenexy" refers to glycyl-L-2-methylprolyl-L-glutamic acid of formula I: I, Each of the three-dimensional centers is presented S Configuration. The IUPAC name for trafinone is (2S)-2-amino]glutaric acid. Trafinone is also known as "G-2-MePE", "H-Gly-MePro-Glu-OH" or "Gly-MePro-Glu-OH".

[0111] Trifenex can form a crystalline solid by incorporating a solubilizer, such as water or methanol, into the crystal lattice without any chemical change in the trafenex molecule. The term "trafenex hydrate" is represented by the formula: trafenex·xH₂O, where x is the molar ratio of H₂O per mole of trafenex. The trafenex hydrate does not necessarily contain stoichiometric amounts of water; for example, x can be about 2.5. In one embodiment, x is about 1 to about 5. In another embodiment, x is about 2 to about 4. In another embodiment, x is about 2.5 to about 3.5. In another embodiment, x is about 2. In another embodiment, x is about 2.5. In another embodiment, x is about 3. In another embodiment, x is about 3.5. In another embodiment, x is about 4. In another embodiment, x is about 4.5.

[0112] As used herein, the term "substantially pure" in relation to trafeneptide polymorphs means that the crystalline material contains about 10% or less by weight, for example about 1% to about 10%, such as about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, or about 1% of any other crystalline or amorphous form of trafeneptide or trafeneptide hydrate. In another embodiment, the trafeneptide polymorph is substantially pure form A.

[0113] As used herein, the term "pure" in relation to trafenexide polymorphs means that the crystalline material contains about 1% or less by weight, for example about 0.1% to about 1%, such as about 1%, about 0.9%, about 0.8%, about 0.7%, about 0.6%, about 0.5%, about 0.4%, about 0.3%, about 0.2%, or about 0.1% or less of any other crystalline or amorphous form of trafenexide. In one embodiment, the crystalline trafenexide polymorph (e.g., form A) does not contain any other crystalline or amorphous form of trafenexide or trafenexide hydrate in a PXRD-detectable amount. In another embodiment, the trafenexide polymorph is pure form A.

[0114] As used in this article, the term "amorphous" refers to a solid form of tricfenestrate or tricfenestrate hydrate that lacks the long-range ordering characteristics of crystals, i.e., the solid is non-crystalline.

[0115] As used herein, the term "substantially identical" regarding PXRD peak positions and / or relative intensities means that peak position and / or intensity variability is taken into account when comparing PXRD diffraction patterns. Similarly, the term "substantially identical" regarding Raman or IR peak positions means that peak position variability is taken into account when comparing Raman or IR spectra. For example, PXRD peak positions can exhibit, for instance, inter-instrument variability, such as up to 0.2°2Θ, or ±0.2 degrees 2Θ; Raman and IR peak positions can exhibit, for instance, inter-instrument variability, such as up to 4 cm. -1 That is, ± 4 cm -1 For example, in PXRD diffraction patterns, relative peak intensities can also show inter-device variability due to crystallinity, orientation, the surface of the prepared sample, and other factors known to those skilled in the art; therefore, they should only be considered qualitative measures.

[0116] As used in this article, the term “micronization” refers to the process or method of reducing the size of a population of particles, typically to the micrometer level.

[0117] As used in this article, the term "micron" or "μm" refers to "micrometer," which is 1 x 10⁻⁶. -6 rice.

[0118] As used herein, the term "therapeutic effective amount" refers to an amount of trafenexylide sufficient to treat one or more symptoms of a disease, condition, injury or symptom, or to prevent the progression of a disease, condition, injury or symptom, or to cause the resolution of a disease, condition, injury or symptom.

[0119] As used herein, the terms "chemically stable" and similar terms for trafeneptide polymorphs mean that after storage at a temperature of about 25°C and a relative humidity of about 60% for at least 3 months, the trafeneptide crystalline solid shows less than 0.5% chemical degradation, for example, less than 0.4%, less than 0.3%, less than 0.2%, less than 0.1%, or less than 0.05% chemical degradation. In determining the amount of degradation, methods known in the art, such as HPLC, can be used to measure the presence of one or more chemical impurities and / or to measure the disappearance of trafeneptide.

[0120] The terms “a” and “a kind” refer to one (kind) or more than one (kind).

[0121] As used in this article, the term “about” includes ±10% of the enumerated number. Therefore, “about 10” means 9 to 11.

[0122] As used in this article, the term "average particle size distribution" or "D" refers to... 50 "The diameter is the diameter of 50% by mass particles with a larger equivalent diameter, as determined by laser diffraction in a Malvern Master Sizer Microplus device or its equivalent device, where the other 50% by mass particles have a smaller equivalent diameter."

[0123] As used herein, the term "excipient" means any component, other than trafeneptide polymorph, that is in or added to a pharmaceutical formulation (e.g., an orally administered solution) suitable for administration to a subject. Excipients are generally inert substances, such as water, added to a composition to facilitate the processing, handling, dissolution, administration, etc., of trafeneptide polymorph. Useful excipients include, but are not limited to, adjuvants, anti-adhesion agents, binders, carriers, disintegrants, fillers, flavoring agents, colorants, diluents, lubricants, flow aids, preservatives, adsorbents, solvents, surfactants, and sweeteners.

[0124] Conventional pharmaceutical excipients are well known to those skilled in the art. A wide variety of pharmaceutical excipients, including water, can be used in mixtures containing trafenexylidene polymorphs. Handbook of Pharmaceutical Excipients Pharmaceutical Press, 4th edition (2003) and Remington: The Science and Practice of PharmacyOther substances listed in Lippincott Williams & Wilkins, 21st edition (2005). In one embodiment, the composition comprises form A dissolved in water.

[0125] As used herein, the term "subject" refers to an animal, such as a human or a veterinary animal, such as a cow, sheep, pig, horse, dog, or cat. In one implementation, the subject is a human.

[0126] As used herein, the term “container” means any storage vessel and closure that is therefore suitable for storing, transporting, dispensing and / or disposing of pharmaceutical products or excipients.

[0127] The term "insert" refers to information accompanying a pharmaceutical product that provides a description of how to administer the product, as well as safety and efficacy data necessary to allow doctors, pharmacists, and patients to make informed decisions about its use. Packaging inserts are often considered the "label" of a pharmaceutical product.

[0128] As used herein, the term “and / or” should be considered as a specific disclosure of each of the two specified features or components in the presence or absence of the other. Thus, the term “and / or” as used herein in phrases such as “A and / or B” is intended to include “A and B”, “A or B”, “A” (alone), and “B” (alone). Similarly, the term “and / or” as used in phrases such as “A, B, and / or C” is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0129] VII. Specific Implementation Plan This disclosure provides the following specific implementation plan.

[0130] Implementation Scheme 1. Crystalline tricfenexide·xH2O, wherein x is about 2 to about 4, characterized by having: (i) Powder X-ray diffraction patterns with peaks at 2Θ degrees 6.8, 11.5, 12.6, 13.8, and 16.8 degrees using Cu Kα radiation, where the 2Θ values ​​are ±0.2 degrees 2Θ; or (ii) Powder X-ray diffraction patterns with d-spacings at 13.1, 7.7, 7.0, 6.4, and 5.3 Å using Cu Kα radiation; or (iii) at 2989, 2934, 2883, 1685, 1637, 1459 and 930 cm -1 The FT-Raman spectrum with peaks at cm -1 The value is ±4 cm -1 ;or (iv) at 13, 24, 67 and 77 cm -1 The low-frequency (LF) Raman spectrum with a peak at cm -1 The value is ±4 cm -1 ;or (v) Peaks at 179.7, 177.9, 177.5, 177.2, 177.0, 165.3, 164.9, 164.8, 67.8, 67.4, 58.6, 58.2, 46.8, 40.3, 33.3, 25.3, 23.5, and 21.1 ppm 13 C solid-state nuclear magnetic resonance spectrum, with ppm values ​​of ±3 pm; or (vi) Having 18 peaks 13 C solid-state NMR spectra, where: (i) the Δ from the farthest low-field peak to the second farthest low-field peak is 1.8 ppm; (ii) the Δ from the farthest low-field peak to the third farthest low-field peak is 2.2 ppm; (iii) the Δ from the farthest low-field peak to the fourth farthest low-field peak is 2.5 ppm; (iv) the Δ from the farthest low-field peak to the fifth farthest low-field peak is 2.7 ppm; (v) the Δ from the farthest low-field peak to the sixth farthest low-field peak is 14.4 ppm; (vi) the Δ from the farthest low-field peak to the seventh farthest low-field peak is 14.8 ppm; (vii) the Δ from the farthest low-field peak to the eighth farthest low-field peak is 14.9 ppm; (viii) the Δ from the farthest low-field peak to the ninth farthest low-field peak is 111.9 ppm; (ix) the Δ from the farthest low-field peak to the tenth farthest low-field peak is 112.3 ppm. ppm; (x) Δ from the farthest low-field peak to the eleventh farthest low-field peak is 121.1 ppm; (xi) Δ from the farthest low-field peak to the twelfth farthest low-field peak is 121.5 ppm; (xii) Δ from the farthest low-field peak to the thirteenth farthest low-field peak is 133.1 ppm; (xiii) Δ from the farthest low-field peak to the fourteenth farthest low-field peak is 139.4 ppm; (xiv) Δ from the farthest low-field peak to the fifteenth farthest low-field peak is 146.3 ppm; (xv) Δ from the farthest low-field peak to the sixteenth farthest low-field peak is 154.6 ppm; (xvi) Δ from the farthest low-field peak to the seventeenth farthest low-field peak is 156.2 ppm; and / or (xvii) Δ from the farthest low-field peak to the farthest high-field peak is 158.6 ppm, or any combination thereof; or (vii) The melting point, based on differential scanning calorimetry, has an initial temperature of 71.71℃ and a peak temperature of 72.06℃; or (viii) at 1678, 1636, 1589, 1525, 1214 and 1196 cm -1 The infrared (IR) spectrum with a peak at a certain point, where cm -1The value is ±4 cm -1 ;or (ix) at 5145, 4630 and 4423 cm -1 The near-infrared (NIR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 ;or Its combination.

[0131] Implementation Scheme 2. Crystalline tricfenexide·xH2O, wherein x is about 2 to about 4, characterized by having: (i) Powder X-ray diffraction patterns with peaks at 6.8, 11.5, 12.6, 13.8, and 16.8 degrees 2Θ using Cu Kα radiation, where the 2Θ values ​​are ±0.2 degrees 2Θ; and / or (ii) Powder X-ray diffraction patterns with d-spacings at 13.1, 7.7, 7.0, 6.4, and 5.3 Å using Cu Kα radiation; and / or (iii) at 2989, 2934, 2883, 1685, 1637, 1459 and 930 cm -1 The FT-Raman spectrum with peaks at cm -1 The value is ±4 cm -1 ; and / or (iv) at 13, 24, 67 and 77 cm -1 The low-frequency (LF) Raman spectrum with a peak at cm -1 The value is ±4 cm -1 ; and / or (v) Peaks at 179.7, 177.9, 177.5, 177.2, 177.0, 165.3, 164.9, 164.8, 67.8, 67.4, 58.6, 58.2, 46.8, 40.3, 33.3, 25.3, 23.5, and 21.1 ppm 13 C solid-state nuclear magnetic resonance spectrum, with ppm values ​​of ±3 pm; and / or (vi) Having 18 peaks 13C solid-state NMR spectra, where: (i) the Δ from the farthest low-field peak to the second farthest low-field peak is 1.8 ppm; (ii) the Δ from the farthest low-field peak to the third farthest low-field peak is 2.2 ppm; (iii) the Δ from the farthest low-field peak to the fourth farthest low-field peak is 2.5 ppm; (iv) the Δ from the farthest low-field peak to the fifth farthest low-field peak is 2.7 ppm; (v) the Δ from the farthest low-field peak to the sixth farthest low-field peak is 14.4 ppm; (vi) the Δ from the farthest low-field peak to the seventh farthest low-field peak is 14.8 ppm; (vii) the Δ from the farthest low-field peak to the eighth farthest low-field peak is 14.9 ppm; (viii) the Δ from the farthest low-field peak to the ninth farthest low-field peak is 111.9 ppm; (ix) the Δ from the farthest low-field peak to the tenth farthest low-field peak is 112.3 ppm. ppm; (x) Δ from the farthest low-field peak to the eleventh farthest low-field peak is 121.1 ppm; (xi) Δ from the farthest low-field peak to the twelfth farthest low-field peak is 121.5 ppm; (xii) Δ from the farthest low-field peak to the thirteenth farthest low-field peak is 133.1 ppm; (xiii) Δ from the farthest low-field peak to the fourteenth farthest low-field peak is 139.4 ppm; (xiv) Δ from the farthest low-field peak to the fifteenth farthest low-field peak is 146.3 ppm; (xv) Δ from the farthest low-field peak to the sixteenth farthest low-field peak is 154.6 ppm; (xvi) Δ from the farthest low-field peak to the seventeenth farthest low-field peak is 156.2 ppm; and / or (xvii) Δ from the farthest low-field peak to the farthest high-field peak is 158.6 ppm, or any combination thereof; and / or (vii) Melting point with an initial temperature of 71.71 °C and a peak temperature of 72.06 °C, based on differential scanning calorimetry; and / or (viii) at 1678, 1636, 1589, 1525, 1214 and 1196 cm -1 The infrared (IR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 ; and / or (ix) at 5145, 4630 and 4423 cm -1 The near-infrared (NIR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 ; and / or Its combination.

[0132] Implementation Scheme 3. Crystalline trefeneptide·xH2O as described in Implementation Scheme 1 or 2, characterized in that it has a powder X-ray diffraction pattern with peaks at 6.8, 11.5, 12.6, 13.8 and 16.8 degrees 2Θ using Cu Kα radiation, wherein the 2Θ value is ±0.2 degrees 2Θ.

[0133] Implementation Scheme 4. The crystalline trefeneptide·xH2O as described in Implementation Scheme 3, characterized by having a powder X-ray diffraction pattern with peaks at 22.3, 23.6, 25.3 and / or 28.1 degrees 2Θ using CuKα radiation, wherein the 2Θ value is ±0.2 degrees 2Θ.

[0134] Implementation Scheme 5. Crystalline trefeneptide·xH2O as described in Implementation Scheme 1 or 2, characterized in that it has a powder X-ray diffraction pattern with peaks at 6.7 or 6.8, 11.4 or 11.5, 12.6 and 13.7 or 13.8 degrees 2Θ using Cu Kα radiation, wherein the 2Θ value is ±0.2 degrees 2Θ.

[0135] Implementation Scheme 6. Crystalline trefeneptide·xH2O as described in Implementation Scheme 1 or 2, characterized by having a powder X-ray diffraction pattern with peaks at 2Θ degrees 6.7 or 6.8, 11.4 or 11.5, 12.6 and 13.7 or 13.8 using Cu Kα radiation.

[0136] Implementation Scheme 7. Crystalline trefeneptide·xH2O as described in Implementation Scheme 5 or 6, characterized in that it has a powder X-ray diffraction pattern with peaks at 22.3, 23.6, 25.3 and / or 28.1 degrees 2Θ ± 0.2 degrees 2Θ using Cu Kα radiation.

[0137] Implementation Scheme 8. Crystalline trefeneptide·xH2O as described in Implementation Scheme 7, characterized by having a powder X-ray diffraction pattern with peaks at 6.7, 11.4, 12.6, 13.7, 22.3, 23.6, 25.3 and 28.1 degrees 2Θ using CuKα radiation, wherein the 2Θ value is ±0.2 degrees 2Θ.

[0138] Implementation Scheme 9. Crystalline trefeneptide·xH2O as described in any one of Implementation Schemes 1-3, characterized in that it has powder X-ray diffraction patterns with d-spacing at 13.1, 7.7, 7.0, 6.4 and 5.3 Å using Cu Kα radiation.

[0139] Implementation Scheme 10. Crystalline trefeneptide·xH2O as described in any one of Implementation Schemes 1-9, characterized by having: [values ​​at 2989, 2934, 2883, 1685, 1637, 1459, and 930 cm⁻¹] -1 The FT-Raman spectrum with peaks at cm -1 The value is ±4 cm -1 .

[0140] Implementation Scheme 11. Crystalline trefeneptide·xH2O as described in any one of Implementation Schemes 1-10, characterized by having: at 13, 24, 67 and 77 cm⁻¹ -1 The low-frequency (LF) Raman spectrum with a peak at cm -1 The value is ±4 cm -1 .

[0141] Scheme 12. Crystalline trefeneptide·xH2O as described in any one of Schemes 1-11, characterized in that it has peaks at 179.7, 177.9, 177.5, 177.2, 177.0, 165.3, 164.9, 164.8, 67.8, 67.4, 58.6, 58.2, 46.8, 40.3, 33.3, 25.3, 23.5 and 21.1 ppm. 13 C solid-state nuclear magnetic resonance spectrum, with ppm values ​​of ±3 pm.

[0142] Implementation Scheme 13. Crystalline tricfenexylidene·xH2O as described in any one of Implementation Schemes 1-12, characterized in that it has: having 18 peaks 13 The C solid-state NMR spectrum shows that: (i) the Δ from the farthest low-field peak to the second farthest low-field peak is 1.8 ppm; (ii) the Δ from the farthest low-field peak to the third farthest low-field peak is 2.2 ppm; (iii) the Δ from the farthest low-field peak to the fourth farthest low-field peak is 2.5 ppm; (iv) the Δ from the farthest low-field peak to the fifth farthest low-field peak is 2.7 ppm; (v) the Δ from the farthest low-field peak to the sixth farthest low-field peak is 14.4 ppm; (vi) the Δ from the farthest low-field peak to the seventh farthest low-field peak is 14.8 ppm; (vii) the Δ from the farthest low-field peak to the eighth farthest low-field peak is 14.9 ppm; and (viii) the Δ from the farthest low-field peak to the ninth farthest low-field peak is 111.9 ppm. ppm; (ix) Δ from the farthest low-field peak to the tenth farthest low-field peak is 112.3 ppm; (x) Δ from the farthest low-field peak to the eleventh farthest low-field peak is 121.1 ppm; (xi) Δ from the farthest low-field peak to the twelfth farthest low-field peak is 121.5 ppm; (xii) Δ from the farthest low-field peak to the thirteenth farthest low-field peak is 133.1 ppm; (xiii) Δ from the farthest low-field peak to the fourteenth farthest low-field peak is 139.4 ppm; (xiv) Δ from the farthest low-field peak to the fifteenth farthest low-field peak is 146.3 ppm; (xv) Δ from the farthest low-field peak to the sixteenth farthest low-field peak is 154.6 ppm; (xvi) Δ from the farthest low-field peak to the seventeenth farthest low-field peak is 156.2 ppm. ppm; and / or (xvii) Δ from the furthest low-field peak to the furthest high-field peak is 158.6 ppm, or any combination thereof.

[0143] Implementation Scheme 14. Crystalline tricfenexylidene·xH2O as described in any one of Implementation Schemes 1-13, characterized in that it has a melting point with an initial temperature of 71.71°C and a peak temperature of 72.06°C based on differential scanning calorimetry.

[0144] Scheme 15. Crystalline trefeneptide·xH2O as described in any one of Schemes 1-14, characterized by a concentration of 1678, 1636, 1589, 1525, 1214, and 1196 cm⁻¹. -1 The infrared (IR) spectrum with a peak at a certain point, where cm -1 The value is ±4cm -1 .

[0145] Implementation Scheme 16. Crystalline trefeneptide·xH2O as described in any one of Implementation Schemes 1-15, characterized in that it is prepared at 5145, 4630, and 4423 cm⁻¹ -1 The near-infrared (NIR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 .

[0146] Implementation Scheme 17. Crystalline tricfenexide·xH2O as described in any one of Implementation Schemes 1-16, having an average particle size distribution of about 10 μm to about 500 μm.

[0147] Implementation Scheme 18. Crystalline tricfenexylidene·xH2O as described in any one of Implementation Schemes 1-17, wherein x is about 2.5 to about 3.5.

[0148] Implementation Scheme 19 Crystallized trefeneptide·xH2O as described in any one of Implementation Schemes 1-17, wherein x is about 2.

[0149] Implementation Scheme 20. Crystalline tricfenexylidene·xH2O as described in any one of Implementation Schemes 1-17, wherein x is about 2.5.

[0150] Implementation Scheme 21. Crystalline tricfenexylidene·xH2O as described in any one of Implementation Schemes 1-17, wherein x is about 3.

[0151] Implementation Scheme 22. Crystalline tricfenexylidene·xH2O as described in any one of Implementation Schemes 1-17, wherein x is about 3.5.

[0152] Implementation Scheme 23. Crystalline tricfenexylidene·xH2O as described in any one of Implementation Schemes 1-17, wherein x is about 4.

[0153] Implementation Scheme 24. A pharmaceutical composition comprising crystalline trefeneptide xH2O as described in any one of Implementation Schemes 1-23 and a pharmaceutically acceptable excipient.

[0154] Implementation Scheme 25. The pharmaceutical composition as described in Implementation Scheme 24 is in particulate form.

[0155] Implementation Scheme 26. An aqueous pharmaceutical preparation comprising crystalline trefeneptide·xH2O as described in any one of Implementation Schemes 1-23 dissolved in water.

[0156] Implementation Scheme 27. An aqueous pharmaceutical preparation as described in Implementation Scheme 26, wherein approximately 1 gram of crystalline trofenapyline·xH2O is dissolved in every 5 mL of water.

[0157] Implementation Scheme 28. A method for preparing an aqueous pharmaceutical formulation as described in Implementation Scheme 26 or 27, the method comprising mixing the crystalline trefeneptide·xH2O with water.

[0158] Implementation Scheme 29. A medicine box comprising crystalline trafeneptide·xH2O as described in any one of Implementation Schemes 1-23 and instructions for dissolving the crystalline trafeneptide·xH2O in water to provide an aqueous pharmaceutical preparation.

[0159] Implementation Scheme 30. The medicine box as described in Implementation Scheme 29, further comprising instructions for administering the aqueous pharmaceutical preparation to a subject suffering from a disease, condition, or symptom.

[0160] Implementation Scheme 31. The medicine box as described in Implementation Scheme 30, wherein the disease, symptom or condition is traumatic brain injury.

[0161] Implementation Scheme 32. The medicine box as described in Implementation Scheme 30, wherein the disease, symptom or condition is a neurodevelopmental disorder.

[0162] Implementation Scheme 33. The medicine box as described in Implementation Scheme 32, wherein the neurodevelopmental disorder is Rett syndrome, fragile X syndrome or autism spectrum disorder.

[0163] Implementation Scheme 34. A method for treating a disease, condition, or symptom in a subject in need, the method comprising administering to the subject a pharmaceutical composition as described in Implementation Scheme 24 or 25 or an aqueous pharmaceutical preparation as described in Implementation Scheme 26 or 27.

[0164] Implementation Scheme 35. The method as described in Implementation Scheme 34, wherein the disease, symptom, or condition is traumatic brain injury.

[0165] Implementation Scheme 36. The method as described in Implementation Scheme 34, wherein the disease, symptom, or condition is a neurodevelopmental disorder.

[0166] Implementation Scheme 37. The method as described in Implementation Scheme 36, wherein the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0167] Implementation Scheme 38. The method as described in Implementation Scheme 34, wherein the disease, symptom, or condition is Rett syndrome.

[0168] Implementation Scheme 39. A pharmaceutical composition as described in Implementation Scheme 24 or 25 or an aqueous pharmaceutical preparation as described in Implementation Scheme 26 or 27, for use in treating a disease, condition or symptom in a subject in need.

[0169] Implementation Scheme 40. The composition or formulation as described in Implementation Scheme 39, wherein the disease, condition or symptom is traumatic brain injury.

[0170] Implementation Scheme 41. The composition or formulation as described in Implementation Scheme 39, wherein the disease, condition or symptom is a neurodevelopmental disorder.

[0171] Implementation Scheme 42. The composition or formulation as described in Implementation Scheme 41, wherein the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0172] Implementation Scheme 43. The composition or formulation as described in Implementation Scheme 39, wherein the disease, condition or symptom is Rett syndrome.

[0173] Implementation Scheme 44. Use of trafenexide as described in any one of Implementation Schemes 1-23, or the pharmaceutical composition as described in Implementation Scheme 24 or 25, or the aqueous pharmaceutical formulation as described in Implementation Scheme 26 or 27 in the manufacture of a medicament for a disease, condition, or symptom of a subject in need.

[0174] Implementation Scheme 45. The use as described in Implementation Scheme 44, wherein the disease, symptom, or condition is traumatic brain injury.

[0175] Implementation Scheme 46. The use as described in Implementation Scheme 44, wherein the disease, symptom, or condition is a neurodevelopmental disorder.

[0176] Implementation Scheme 47. Use as described in Implementation Scheme 46, wherein the neurodevelopmental disorder is Rett syndrome, Fragile X syndrome, or autism spectrum disorder.

[0177] Implementation Scheme 48. The use as described in Implementation Scheme 44, wherein the disease, symptom, or condition is Rett syndrome.

[0178] Implementation Scheme 49. A method for preparing crystalline trafeneptide·xH2O as described in any one of Implementation Schemes 1-23, the method comprising i) adding ethanol to an aqueous solution of trafeneptide at about 25°C; ii) cooling the solution to about 0°C; and iii) separating the solid thereby obtained to obtain crystalline trafeneptide·xH2O.

[0179] Implementation Scheme 50. The method as described in Implementation Scheme 49, wherein the ratio of water to ethanol is approximately 3:7 w / w.

[0180] Example instrument Powder X-ray diffraction (PXRD or XRPD) PXRD and XRPD are synonymous terms. The Rigaku Smart-Lab X-ray diffraction system is configured to reflect a Bragg-Brentano geometry using a line-source X-ray beam. The X-ray source is a Cu Long Fine Focus tube (λ = 1.54 Å) operating at 40 kV and 44 mA. The source provides an incident beam profile on the sample ranging from a narrow line at high angles to a wide rectangle at low angles. A beam-adjusting slit is used on the line X-ray source to ensure that the maximum beam size along and perpendicular to the line is less than 10 mm. The Bragg-Brentano geometry is a para-focusing geometry controlled by passive divergence and receiving slits, with the sample itself acting as the focusing element of the optics. The inherent resolution of the Bragg-Brentano geometry depends in part on the diffractometer radius and the width of the receiving slit used. Typically, the Rigaku Smart-Lab is operated to provide a peak width of 0.1°2Θ or less. The axial divergence of the X-ray beam is controlled by a 5.0-degree Soller slit in both the incident and diffracted beam paths.

[0181] Powder samples were prepared in a low-background Si holder using slight manual compaction to keep the sample surface flat and flush with the reference surface of the sample holder. Each sample was analyzed using continuous scans at 6°2Θ per minute, from 2 to 40°2Θ, with an effective step size of 0.02°2Θ.

[0182] Differential scanning calorimetry (DSC) DSC analysis was performed using a TA Instruments Q2500 Discovery series instrument. Instrument temperature calibration was performed using indium. During the analysis, the DSC cell was purged with nitrogen at approximately 50 mL per minute. The sample was placed in a standard rolled aluminum tray and heated from approximately 25°C to 350°C at a rate of 10°C per minute.

[0183] Dynamic vapor adsorption (DVS) analysis DVS analysis was performed using a TA Instruments Q5000 dynamic vapor adsorption analyzer. This instrument was calibrated using standard weights and a sodium bromide humidity standard. Samples were analyzed in 10% relative humidity (RH) steps from 5% to 95% RH (adsorption cycle) and from 95% to 5% RH (desorption cycle) at 25°C (maximum equilibration time 60 minutes).

[0184] Infrared (IR) spectroscopy Infrared spectra were obtained on a Nicolet 6700 FT-IR system using a Nicolet SMART iTR attenuated total reflection device.

[0185] Near-infrared (NIR) spectroscopy Near-infrared spectra were obtained on a Nicolet iS50 IR system. Approximately 1% w / w of trefeneptide A in dry KBr was placed in a DRIFT (diffuse reflectance Fourier transform infrared spectroscopy) cuvette and analyzed in the spectral range of 8000 and 400 wavenumbers.

[0186] FT-Raman spectroscopy Fourier transform (FT) Raman spectra were acquired on a Nicolet 6700 spectrometer connected to the Nexus Raman Attachment Module. This instrument was configured with an Nd:YAG laser operating at 1024 nm, a CaF2 beam splitter, and an indium gallium arsenide detector. OMNIC 8.1 software was used to control data acquisition and spectral processing. Samples were placed in 3-inch glass NMR tubes for analysis.

[0187] Low-frequency (LF) Raman spectroscopy Low-frequency Raman spectra were obtained using a Renishaw inVia Raman microscope equipped with an Ondax THz-Raman system (TR-PROBE; excitation laser 853.1 nm, notch filter). Sample powder was analyzed in the open air using a probe tip attachment. The microscope was operated at a 36 cm⁻¹ aperture. -1 Static scan centered on the spectral range of -575 to 575 cm⁻¹ -1 Within the range, spectra were acquired at 100% power, a one-second exposure time, and 32 cumulative exposures. Wavelength calibration was verified using a sulfur reference standard. Data acquisition was performed using WiRE 3.4 software.

[0188] Karl Fischer (KF) analysis Karl Fischer analysis was performed using a Mettler-Toledo C20 Coulometric KF titrator. The instrument was calibrated using a Hydranal water standard containing 1% water. The titrant was a Hydranal methanol solution.

[0189] 13 solid-state nuclear magnetic resonance (NMR) spectroscopy Solid-state experiments were conducted on a Bruker Avance II 400 spectrometer equipped with a Doty probe (DSI-1630) 1H (19F) / X dual resonance. 13 Cross-polarization magic angle rotation (CPMAS) experiment. The sample (109 mg) was loaded into a 4-mm 4mm silicon nitride rotator sealed with a Kel-F end cap for subsequent data acquisition. Adamantane was set to have a methylene signal of 38.48 ppm on the TMS scale and was used as an external standard. The acquisition and processing parameters used are shown in the table below. It is possible to use different magnetic fields (such as 9.4 Tesla). 13 C is 100 MHz. 1 (H is 400 MHz or higher), performed on an NMR spectrometer. 13 C CPMAS analysis. Parameters such as acquisition time, dwell time, cycle delay, rotation speed, and number of scans can be modified and optimized according to the NMR spectrometer.

[0190] Single crystal structure determined X-ray diffraction analysis was performed at 150 K using Cu Kα radiation (λ = 1.54 Å). Equation C 13 H 21 The monoclinic cell parameters and calculated volume of N3O6·3(H2O) are a = 18.8946 (8) Å, b = 7.2849 (3) Å, c = 27.8601 (12) Å, β = 109.8540 (16) Å. And V = 3606.9 (3) Å 3 .for Z =8 and formula weight 369.37, the calculated density is 1.360 g / cm³. 3 .

[0191] Example 1 Synthesis and characterization of form A Form A is prepared according to the following method.

[0192] Method 1 19.9 mg of amorphous tricfenexylidene was loaded into a mechanical grinding vessel containing a metal ball. 7.6 mg of L-asparagine and 10 μL of water were added to the grinding vessel. The vessel was sealed and ground on a Retsch Mill at 100% power for approximately 20 minutes. The resulting solid was removed and dried overnight in a vacuum desiccator. XRPD analysis revealed that the material was a mixture of form A and crystalline L-aspartic acid.

[0193] Method 2 18.6 mg of amorphous tricfenex was loaded into a mechanical grinding vessel with a metal ball. 8.0 mg of L-aspartic acid and 10 μL of water were added to the grinding vessel. The vessel was sealed and ground on a Retsch Mill for approximately 20 minutes. The resulting solid was allowed to dry in an open container, after which it was transferred. XRPD analysis revealed that the material was a mixture of form A and crystalline L-aspartic acid. The same experiment was repeated using 20 mg of amorphous tricfenex and 1 mg of L-aspartic acid. According to XRPD, the resulting material was predominantly form A with trace amounts of crystalline L-aspartic acid. This material was used as seed crystals (3 mg) in a repeat experiment using only amorphous tricfenex (19.7 mg). According to XRPD, the resulting solid was form A.

[0194] Method 3 At ambient temperature, a trafenexyline solution (300 mL, 32% w / w trafenexyline amorphous form in water) was added to anhydrous ethanol (1200 mL). The resulting solution was cooled to 2°C with stirring (300 rpm). The solution was reheated to 25°C (12°C / hr, held for 3 hours) and cooled to 2°C (6°C / hr). The precipitated solid was filtered, washed with cold ethanol (2°C), and dried under nitrogen at 70% RH / ambient temperature for 16 hours to remove residual ethanol. According to XRPD, the resulting solid was form A.

[0195] Method 4 Form A is prepared using the following steps according to method 4.

[0196] At room temperature, 205.7 kg of amorphous tricfenexylidene (KF 5.0%, 195.4 kg dry basis) was dissolved in 617 kg of water in a reactor.

[0197] At room temperature, 2808 kg of ethanol was added to the solution in the reactor under vigorous stirring.

[0198] Cool the solution to 0-2℃.

[0199] 1.0 kg of trofenapyline seed crystals were added to the reactor, and this batch was aged NLT for 6 hours with slow stirring. Crystallization will occur.

[0200] The slurry in the filter reactor is then separated in a filter dryer.

[0201] Wash the wet filter cake twice with 325 kg of ethanol pre-cooled to 0-2℃.

[0202] The wet filter cake was vacuum dried until most of the residual solvent was evaporated, and then the jacket temperature of the filter dryer was raised to room temperature to complete the drying. 170.0 kg of crystalline tricfenex·xH2O (KF 13.5%, 147.0 kg dry basis) was separated (75% yield).

[0203] Method 5 Form A is prepared using the following steps according to method 5.

[0204] At room temperature, ethanol (4479 kg) was added to a trafeneptide aqueous solution (total weight 1272 kg, 23.2 w / w% trafeneptide, 294.5 kg trafeneptide, 977.5 kg water) under vigorous mixing.

[0205] Cool the solution to 0-2℃.

[0206] Add 2.5 kg of crystalline trefeneptide·xH2O seed crystals to the reactor and age this batch of NLT for 6 hours with slow stirring. Crystallization will occur.

[0207] The slurry in the filter reactor is then separated in a filter dryer.

[0208] Wash the wet filter cake twice with 502 kg of ethanol pre-cooled to 0-2℃.

[0209] The wet filter cake was vacuum dried until most of the residual solvent was evaporated, and then the jacket temperature of the filter dryer was raised to room temperature to complete the drying. 292.0 kg of crystalline tricfenex·xH2O (KF 14.3%, 250.2.0 kg dry basis) was separated (85% yield).

[0210] Method 6 Form A is prepared using the following steps according to method 6.

[0211] At room temperature, 58.1 g of crystalline tricfenexylide·xH2O (KF 14.0%, 50.0 g dry basis) was dissolved in 100 g of water in reactor 1.

[0212] At room temperature, with vigorous stirring, 233 g (296 mL) of ethanol was added to the solution in reactor 1. (The water / ethanol ratio was approximately 3 / 7 w / w, and the target solution concentration was approximately 15% w / w trafenexide).

[0213] 128 g of solution in reactor 1 was transferred to reactor 2 (approximately 1 / 3 of the solution).

[0214] Cool reactor 2 to 0-2℃.

[0215] A slurry was prepared by mixing 0.5 g of trefenapeptide seed material in 5 g ethanol / water (95 / 5 w / w).

[0216] Trifenapyline seed slurry was added to reactor 2 and aged for 2 hours with slow stirring. Nucleation will occur to form a seed bed.

[0217] After two hours of NLT, the remaining solution was transferred from reactor 1 to reactor 2, while reactor 2 was maintained at 0-2°C with good mixing.

[0218] 334 g (423 mL) of ethanol was loaded into reactor 1 and cooled to 0-2℃.

[0219] Ethanol was transferred from reactor 1 to reactor 2 over a NLT period of 2 hours, while reactor 2 was maintained at 0-2°C with good mixing (the final water / ethanol ratio was approximately 15 / 85 w / w).

[0220] Under good mixing conditions and at 0-2℃, the slurry in reactor 2 shall be aged for no less than 2 hours.

[0221] A filter cake washing solution was prepared by mixing 25 g of water and 475 g of EtOH and cooling to 0-2 °C.

[0222] The slurry in reactor 2 is filtered, and then the wet filter cake on the filter is washed at 0-2°C.

[0223] The wet filter cake was vacuum dried at 0–2 °C until most of the residual solvent was evaporated, and then the batch was brought to room temperature to complete the drying. 53.2 g of crystalline tricfenexylide·xH2O (KF 13.5%, 46.0 g dry basis) was isolated (92% yield).

[0224] The structure of form A was resolved by single-crystal X-ray diffraction. The structure shows that each trofenesin molecule contains three water molecules. The asymmetric unit of form A is shown in... Figure 8In the diagram, hydrogen atoms have been omitted for clarity. The structure of form A shows that each of the three water molecules forms a hydrogen bond with either the oxygen or nitrogen atom of the trafinone peptide. Each of the three water molecules also forms a hydrogen bond with its adjacent water molecule.

[0225] The diffraction pattern of form A X-ray powder diffraction (XRPD) is shown in the figure. Figure 1 The XRPD peak list (±0.2 degrees 2Θ) is provided in Table 1.

[0226] Table 1 The Raman spectrum of form A is shown in Figure 2 List of Raman peaks (±4 cm) -1 (This information is provided in Table 2.)

[0227] Table 2 The low-frequency (LF) Raman spectrum of form A is shown in Figure 3 List of LF Raman peaks (±4 cm) -1 (This information is provided in Table 3.)

[0228] Table 3 Form A 13 The solid-state nuclear magnetic resonance (ssNMR) spectrum of 1200 C24 is shown in... Figure 4 The list of ssNMR peaks is provided in Table 4. Selected peaks (with Δppm) are provided in Tables 5 and 6.

[0229] Table 4 Table 5 Table 6 Based on DSC analysis, form A melts at an initial temperature of 71.71 °C and a peak temperature of 72.06 °C. See also... Figure 5 .

[0230] The infrared (IR) spectrum of form A is shown in Figure 6 IR peak list (±4 cm) -1 (This information is provided in Table 7.)

[0231] Table 7 The near-infrared (NIR) spectrum of form A is shown in Figure 7 List of NIR peaks (±4 cm)-1 (This information is provided in Table 8.)

[0232] Table 8 Form A is non-hygroscopic. Dynamic vapor adsorption-desorption (DVS) analysis of form A shows that moisture absorption is minimal when the material is exposed to 5% RH to 95% RH; and moisture loss is also minimal when the material is exposed to 95% RH to 5% RH. Figure 9 According to XRPD, the material obtained after DVS is still in the form of trafinone peptide A (…). Figure 10 ).

[0233] Form A exhibits a 12-14% weight loss between approximately 50 and 120°C. This is likely due to water loss and matches the 12-14% moisture content measured by Karl Fisher analysis. An example TGA scan is presented... Figure 12 In the middle. The DSC data for Form A consistently show a sharp endothermic reaction at approximately 70-72°C (peak temperature), which corresponds to melting.

[0234] Based on the single-crystal X-ray structure showing that each trofenapyline molecule contains three water molecules, form A is a trihydrate ( Figure 8 However, the water content of the generated form A varies between about 12% and about 14%. This indicates that while some water is lattice-essential, at least one water molecule may be loosely bound and can be removed without altering the lattice. For this purpose, form A is designated as trefeneptide·xH2O, where x is about 2 to about 4.

[0235] Example 2 Stability of form A Form A is stable under a wide range of humidity conditions. Form A exposed for two days at 33% RH, 59% RH, 75% RH, and 97% RH showed a molar equivalent of water between 2 and 4. Under extremely dry conditions (open container, exposed for two days at 0% RH), Form A lost water and became disordered. The Form A signal was still visible in the XRPD plot, but the crystallization signal was broad, and the XRPD plot showed some amorphous halos in the baseline, indicating the formation of disordered and amorphous material. Figure 11 ).

[0236] The long-term (6 months) chemical stability of form A and amorphous tricfenex was tested under the same conditions: 25 ± 2 °C / 60 ± 5% relative humidity (RH). Under these conditions, form A was unexpectedly more stable than amorphous tricfenex (Table 9).

[0237] Table 9 PLOQ = Pooled Limit of Quantification; NT = Not Detected; RH = Relative Humidity The analytical methods used in the determination of impurities are provided in Table 10.

[0238] Table 10 It should be understood that the embodiments and examples described above are not intended to limit the scope of this disclosure in any way, and the claims set forth in this disclosure are intended to cover all embodiments and examples, whether or not they are expressly shown herein.

[0239] All patents and publications cited in this article are incorporated herein by reference in their entirety.

Claims

1. Crystalline quaternary peptide·xH2O, wherein x is from about 2 to about 4, characterized in that... have: (i) Powder X-ray diffraction pattern using Cu Kα radiation, showing peaks in the range of 6.6–6.8 degrees 2Θ, 11.3–11.6 degrees 2Θ, 12.5–12.7 degrees 2Θ, and 13.6–13.8 degrees 2Θ, where the 2Θ values ​​are ±0.2 degrees 2Θ; or (ii) Powder X-ray diffraction patterns with d-spacings at 13.1, 7.7, 7.0, 6.4, and 5.3 Å using Cu Kα radiation; or (iii) at 2989, 2934, 2883, 1685, 1637, 1459 and 930 cm -1 The FT-Raman spectrum with peaks at cm -1 The value is ±4 cm -1 ;or (iv) at 13, 24, 67 and 77 cm -1 The low-frequency (LF) Raman spectrum with a peak at cm -1 The value is ±4 cm -1 ;or (v) Peaks at 179.7, 177.9, 177.5, 177.2, 177.0, 165.3, 164.9, 164.8, 67.8, 67.4, 58.6, 58.2, 46.8, 40.3, 33.3, 25.3, 23.5, and 21.1 ppm 13 C solid-state nuclear magnetic resonance spectrum, where the ppm value is ±3 pm; or (vi) Having 18 peaks 13 The C solid-state NMR spectrum shows that: (i) the Δ from the farthest low-field peak to the second farthest low-field peak is 1.8 ppm; (ii) the Δ from the farthest low-field peak to the third farthest low-field peak is 2.2 ppm; (iii) the Δ from the farthest low-field peak to the fourth farthest low-field peak is 2.5 ppm; (iv) the Δ from the farthest low-field peak to the fifth farthest low-field peak is 2.7 ppm; (v) the Δ from the farthest low-field peak to the sixth farthest low-field peak is 14.4 ppm; (vi) the Δ from the farthest low-field peak to the seventh farthest low-field peak is 14.8 ppm; (vii) the Δ from the farthest low-field peak to the eighth farthest low-field peak is 14.9 ppm; and (viii) the Δ from the farthest low-field peak to the ninth farthest low-field peak is 111.9 ppm. ppm; (ix) Δ from the farthest low-field peak to the tenth farthest low-field peak is 112.3 ppm; (x) Δ from the farthest low-field peak to the eleventh farthest low-field peak is 121.1 ppm; (xi) Δ from the farthest low-field peak to the twelfth farthest low-field peak is 121.5 ppm; (xii) Δ from the farthest low-field peak to the thirteenth farthest low-field peak is 133.1 ppm; (xiii) Δ from the farthest low-field peak to the fourteenth farthest low-field peak is 139.4 ppm; (xiv) Δ from the farthest low-field peak to the fifteenth farthest low-field peak is 146.3 ppm; (xv) Δ from the farthest low-field peak to the sixteenth farthest low-field peak is 154.6 ppm; (xvi) Δ from the farthest low-field peak to the seventeenth farthest low-field peak is 156.2 ppm. ppm; and / or (xvii) A is 158.6 ppm from the furthest low-field peak to the furthest high-field peak, or any combination thereof; or (vii) The melting point, based on differential scanning calorimetry, has an initial temperature of 71.71℃ and a peak temperature of 72.06℃; or (viii) at 1678, 1636, 1589, 1525, 1214 and 1196 cm -1 The infrared (IR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 ;or (ix) at 5145, 4630 and 4423 cm -1 The near-infrared (NIR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 ;or Its combination.

2. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... It has a powder X-ray diffraction pattern with peaks at 2Θ degrees of 6.7 or 6.8, 11.4 or 11.5, 12.6 and 13.7 or 13.8 degrees using Cu Kα radiation, where the 2Θ value is ±0.2 degrees of 2Θ.

3. The crystalline trefeneptide·xH2O as described in claim 2, characterized in that... It has powder X-ray diffraction patterns with peaks at 2Θ degrees of 6.7, 11.4, 12.6, 13.7, 22.3, 23.6, 25.3 and 28.1 degrees, using Cu Kα radiation, where the 2Θ values ​​are ±0.2 degrees of 2Θ.

4. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... It has powder X-ray diffraction patterns with d-spacing at 13.1, 7.7, 7.0, 6.4 and 5.3 Å using Cu Kα radiation.

5. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... Featuring: 2989, 2934, 2883, 1685, 1637, 1459 and 930 cm -1 FT-Raman spectra with peaks at, wherein cm -1 The value is ±4 cm -1 .

6. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... Available in sizes of 13, 24, 67, and 77 cm. -1 The low-frequency (LF) Raman spectrum with a peak at cm -1 The value is ±4 cm -1 .

7. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... It has peaks at 179.7, 177.9, 177.5, 177.2, 177.0, 165.3, 164.9, 164.8, 67.8, 67.4, 58.6, 58.2, 46.8, 40.3, 33.3, 25.3, 23.5, and 21.1 ppm. 13 C solid-state nuclear magnetic resonance spectrum, wherein the ppm value is ±3 pm.

8. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... It has a melting point based on differential scanning calorimetry, with an initial temperature of 71.71℃ and a peak temperature of 72.06℃.

9. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... It has the following dimensions: 1678, 1636, 1589, 1525, 1214, and 1196 cm. -1 The infrared (IR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 .

10. The crystalline trefeneptide·xH2O as described in claim 1, characterized in that... Features: in 5145, 4630 and 4423cm -1 The near-infrared (NIR) spectrum with a peak at a certain point, where cm -1 The value is ±4 cm -1 .

11. The crystalline trefeneptide·xH2O as described in claim 1, wherein x is about 2.5 to about 3.

5.

12. The crystalline trefeneptide·xH2O as described in claim 1, wherein x is about 3.

13. A pharmaceutical composition comprising crystalline trefeneptide·xH2O as described in claim 1 and a pharmaceutically acceptable excipient.

14. An aqueous pharmaceutical preparation comprising crystalline trefeneptide·xH2O as described in claim 1 dissolved in water.

15. A method for preparing an aqueous pharmaceutical formulation as claimed in claim 1, the method comprising mixing the crystalline tricfenexide·xH2O with water.

16. A medicine box comprising crystalline trafeneptide·xH2O as claimed in claim 1 and instructions for dissolving crystalline trafeneptide·xH2O in water to provide an aqueous pharmaceutical preparation.

17. A method of treating a disease, condition, or symptom in a subject in need, the method comprising administering to the subject the pharmaceutical composition of claim 13, wherein the disease, condition, or symptom is traumatic brain injury, neurodevelopmental disorder, Rett syndrome, fragile X syndrome, or autism spectrum disorder.

18. The method of claim 17, wherein the disease, symptom, or condition is Rett syndrome.

19. A method for preparing crystalline trafeneptide·xH2O as claimed in claim 1, the method comprising i) adding ethanol to an aqueous solution of trafeneptide at about 25°C; ii) cooling the solution to about 0°C; and iii) separating the solid thereby obtained to obtain crystalline trafeneptide·xH2O.

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