Crystalline forms of trophinetide

Abstract

The present disclosure provides crystalline forms of trofinetide and trophinetide hydrate, pharmaceutical compositions comprising crystalline forms of trofinetide and trophinetide hydrate, methods of making crystalline forms of trofinetide or trophinetide hydrate, and methods of treating a disease, condition, or disorder in a subject comprising administering to the subject a composition comprising a crystalline form of trophinetide or trophinetide hydrate.

Description

[Technical field]

[0001] The present disclosure provides crystalline forms of trofinetide and trophinetide hydrate, pharmaceutical compositions comprising crystalline forms of trofinetide and trophinetide hydrate, methods of making crystalline forms of trofinetide or trophinetide hydrate, and methods of treating a disease, condition, or disorder in a subject comprising administering to the subject a composition comprising a crystalline form of trophinetide or trophinetide hydrate. [Background technology]

[0002] Glycyl-L-2-methylprolyl-L-glutamic acid (also known as trophinetide) is a synthetic analogue of glycine-proline-glutamate (also known as glypromate or GPE). GPE occurs naturally in the brain. It is the N-terminal tripeptide of the insulin-like growth factor 1 (IGF-1) protein. US 7,041,314 discloses trophinetide, a method for making trophinetide, and a method for using trophinetide to treat diseases, disorders, or conditions, such as neurodegeneration caused by hypoxia-ischemia or toxic injury. US 7,605,177 discloses a method for using trophinetide to treat diseases, such as neurodegeneration and chronic neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, diabetic neuropathy caused by type I or type II diabetes, autoimmune diseases of the brain, or multiple sclerosis. US 7,714,020 discloses a method for using trophinetide to treat diseases, disorders, or conditions, such as brain injury caused by traumatic brain injury, stroke, hypoxia / ischemia, and toxic injury. US 7,863,304 discloses a method of using trophinetide to treat a disease, disorder, or condition, such as a chronic neurodegenerative disorder, such as Parkinson's disease. US 8,637,567 discloses a method of using trophinetide to treat a disease, disorder, or condition, such as a cognitive or memory disorder. US 7,887,839, US 8,178,125, and US 8,496,963 disclose oral formulations of trophinetide for treating various diseases, disorders, or conditions. US 9,708,366 and US 9,212,204 disclose a method of using trophinetide to treat a disease, disorder, or condition, such as an autism spectrum disorder, such as autism, autistic disorder Asperger syndrome, childhood disintegrative disorder, pervasive developmental disorder not otherwise specified (PDD-NOS), fragile X syndrome, or Rett syndrome. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] U.S. Patent No. 7,041,314 [Patent Document 2] U.S. Patent No. 7,605,177 [Patent Document 3] U.S. Patent No. 7,714,020 [Patent Document 4] U.S. Patent No. 7,863,304 [Patent Document 5] U.S. Pat. No. 8,637,567 [Patent Document 6] U.S. Pat. No. 7,887,839 [Patent Document 7] U.S. Patent No. 8,178,125 [Patent Document 8] U.S. Pat. No. 8,496,963 [Patent Document 9] U.S. Pat. No. 9,708,366 [Patent Document 10] U.S. Pat. No. 9,212,204 Summary of the Invention

[0004] There is a need for chemically stable solid forms of trophinetide for use in treating Rett Syndrome, Fragile X Syndrome, traumatic brain injury, and other diseases, disorders, and conditions in subjects.

[0005] In one aspect, the present disclosure provides crystalline polymorphic forms of trofinetide or trofinetide hydrate.

[0006] In another aspect, the present disclosure provides methods of making crystalline polymorphic forms of trophinetide.

[0007] In another aspect, the present disclosure provides compositions comprising a crystalline polymorphic form of trofinetide or trofinetide hydrate and one or more excipients.

[0008] In another aspect, the disclosure provides a method of making a composition comprising a crystalline polymorphic form of trofinetide or trophinetide hydrate and one or more excipients.

[0009] In another aspect, the disclosure provides methods of treating a disease, disorder, or condition in a subject, such as traumatic brain injury or a neurodevelopmental disorder, using crystalline polymorphic forms of trophinetide or trophinetide hydrate.

[0010] In another aspect, the present disclosure provides kits comprising crystalline polymorphic forms of trofinetide or trophinetide hydrate. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 is an XRPD diffractogram of Form A. [Diagram 2] 1 is a Raman spectrum of Form A. [Diagram 3] LF-Raman spectrum of Form A. [Figure 4] 1 is a ssNMR spectrum of Form A. [Diagram 5] 1 is a DSC thermogram of Form A. [Figure 6] 1 is an IR spectrum of Form A. [Figure 7] 1 is a NIR spectrum of Form A. [Figure 8] Single crystal X-ray diffraction asymmetric unit of Form A. Hydrogen atoms have been omitted for clarity. [Figure 9] 1 is a line graph showing dynamic vapor sorption / desorption data for Form A. [Figure 10] FIG. 1 is a series of three XRPD diffractograms of Form A containing different amounts of water. [Figure 11] FIG. 1 is a series of five XRPD diffractograms of Form A stored at different humidity conditions. [Figure 12] 1 is a TGA / DCS thermogram of Form A. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] I. Crystalline Trofinetide or Trofinetide Hydrate In one embodiment, the present disclosure provides crystalline polymorphic forms of trofinetide or crystalline polymorphic forms of trofinetide hydrate, collectively referred to as "trophinetide polymorphs."

[0013] In another embodiment, the trophinetide polymorph has the formula: trophinetide·xH2 O, where x is from about 2 to about 4. In another embodiment, the trophinetide polymorph is trophinetide·xH 2 O, where x is from about 2.5 to about 3.5. In another embodiment, the trophinetide polymorph is trophinetide·xH 2 O, where x is about 2. In another embodiment, the trophinetide polymorph is trophinetide·xH 2 O, where x is about 2.5. In another embodiment, the trophinetide polymorph is trophinetide·xH 2 O, where x is about 3. In another embodiment, the trophinetide polymorph is trophinetide·xH 2 O, where x is about 3.5. In another embodiment, the trophinetide polymorph is trophinetide·xH 2 O, where x is about 4. These crystalline trophinetide xH 2 The O polymorphs are collectively referred to as "Form A."

[0014] In another embodiment, Form A is characterized by having a PXRD pattern, i.e., an X-ray powder diffraction pattern, using Cu Kα radiation with peaks in the range of 6.6-6.8 degrees 2θ, a peak in the range of 11.3-11.6 degrees 2θ, a peak in the range of 12.5-12.7 degrees 2θ, and a peak in the range of 13.6-13.8 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ, and optionally with peaks at 16.8, 22.3, 23.6, 25.3, and / or 28.1 degrees 2θ ±0.2 degrees 2θ using Cu Kα radiation.

[0015] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation having 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.

[0016] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with peaks at 6.7, 11.4, 12.6, 13.7, 22.3, 23.6, 25.3, and 28.1 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ.

[0017] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with peaks at 6.7, 11.4, 12.6, 13.7, 22.3, 23.6, 25.3, and 28.1 degrees 2θ.

[0018] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with peaks at 6.8, 11.5, 12.6, 13.8, and 16.8 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ.

[0019] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with peaks at 11.5, 12.6, and 13.8 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ.

[0020] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with peaks at 6.8, 11.5, and 12.6 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ.

[0021] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with 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, with 2θ values ​​of ±0.2 degrees 2θ.

[0022] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with 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, with 2θ values ​​of ±0.2 degrees 2θ.

[0023] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with 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 with 2θ values ​​of ±0.2 degrees 2θ.

[0024] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with 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 with 2θ values ​​of ±0.2 degrees 2θ.

[0025] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with 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 with 2θ values ​​of ±0.2 degrees 2θ.

[0026] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with 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 with 2θ values ​​of ±0.2 degrees 2θ.

[0027] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with 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, with 2θ values ​​of ±0.2 degrees 2θ.

[0028] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least 10 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, with 2θ values ​​of ±0.2 degrees 2θ.

[0029] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with peaks at 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 2θ, with 2θ values ​​of ±0.2 degrees 2θ.

[0030] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with d-spacings of 13.1, 7.7, 7.0, 6.4, and 5.3 Å.

[0031] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with d-spacings of 13.1, 7.7, and 7.0 Å.

[0032] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with d-spacings of 7.7, 7.0, and 6.4 Å.

[0033] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least three d-spacings of 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.

[0034] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least four d-spacings of 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.

[0035] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least five d-spacings of 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.

[0036] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least six d-spacings of 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.

[0037] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least seven d-spacings of 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.

[0038] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least eight d-spacings of 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.

[0039] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least nine d-spacings of 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.

[0040] In another embodiment, Form A is characterized by having a PXRD pattern using Cu Kα radiation with at least ten d-spacings of 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.

[0041] In another embodiment, Form A is characterized by having a PXRD diffractogram essentially the same as that shown in FIG.

[0042] In another embodiment, Form A has the molecular weights of 2989, 2934, 2883, 1685, 1637, 1459, and 930 cm -1 FT-Raman spectrum having a peak at cm -1 Value is ±4cm -1 It is characterized by having an FT-Raman spectrum:

[0043] In another embodiment, Form A has the following molecular weights: 2989, 2960, 2934, 2883, 1685, 1637, 1459, 1417, 1346, 1272, 1199, 1058, 1023, 967, 930, 782, 552, 496, 425, and 342 cm -1 FT-Raman spectrum having a peak at cm -1 Value is ±4cm -1 It is characterized by having an FT-Raman spectrum:

[0044] In another embodiment, Form A is characterized by having an FT-Raman spectrum essentially the same as that shown in FIG.

[0045] In another embodiment, Form A has 13, 24, 67, and 77 cm -1 A low frequency (LF) Raman spectrum with a peak at cm -1 It is characterized by having a low frequency (LF) Raman spectrum with the following values ​​±: 4cm -1 .

[0046] In another embodiment, Form A has the following molecular weights: 13, 24, 34, 67, 77, 208, 283, 348, 422, 495, and 552 cm -1 LF Raman spectrum having a peak at cm -1 Value is ±4cm -1 It is characterized by having an LF Raman spectrum in which

[0047] In another embodiment, Form A is characterized by having an LF-Raman spectrum essentially the same as that shown in FIG.

[0048] In another embodiment, Form A 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 The compound is characterized by having a C solid-state nuclear magnetic resonance (ssNMR) spectrum, the ssNMR spectrum having ppm values ​​of ±3 ppm.

[0049] In another embodiment, Form A has a Δ from the most downfield peak of (i) the second most downfield peak of 1.8 ppm, (ii) the third most downfield peak of 2.2 ppm, (iii) the fourth most downfield peak of 2.5 ppm, (iv) the fifth most downfield peak of 2.7 ppm, (v) the sixth most downfield peak of 14.4 ppm, (vi) the seventh most downfield peak of 14.8 ppm, (vii) the eighth most downfield peak of 14.9 ppm, (viii) the ninth most downfield peak of 111.9 ppm, (ix) the tenth most downfield peak of 112.3 ppm, (x) the eleventh most downfield peak of 113.2 ppm, (y) the tenth most downfield peak of 114.2 ppm, (z) the tenth most downfield peak of 115.2 ppm, (z) the tenth most downfield peak of 116.2 ppm, (z) the tenth most downfield peak of 117.2 ppm, (z) the tenth most downfield peak of 118.2 ppm, (z) the tenth most downfield peak of 119.2 ppm, (z) the tenth most downfield peak of 120.2 ppm, (z) the tenth most downfield peak of 122.2 ppm, (z) the tenth most downfield peak of 123.2 ppm, (z) the tenth most downfield peak of 124.2 ppm, (z) the tenth most downfield peak of 125.2 ppm, (z) the tenth most downfield peak of 126.2 ppm, (z) the tenth most downfield peak of 127.2 ppm, (z) the tenth most downfield peak of 128.2 ppm, (z) the tenth most downfield peak of 129.2 ppm, (z) the tenth most downfield peak of 130.2 ppm, (z) the tenth most downfield peak of (xi) a Δ to the 12th most downfield peak is 121.5 ppm, (xii) a Δ to the 13th most downfield peak is 133.1 ppm, (xiii) a Δ to the 14th most downfield peak is 139.4 ppm, (xiv) a Δ to the 15th most downfield peak is 146.3 ppm, (xv) a Δ to the 16th most downfield peak is 154.4 ppm, (xvi) a Δ to the 17th most downfield peak is 156.2 ppm, and / or (xvii) a Δ from the most downfield peak to the most upfield peak is 158.6 ppm, or any combination thereof. See, e.g., Tables 5 and 6.

[0050] In another embodiment, Form A is characterized by having a ssNMR spectrum essentially the same as that shown in FIG.

[0051] In another embodiment, Form A is characterized by having a melting point with an onset temperature of 71.71° C. and a peak temperature of 72.06° C. based on Differential Scanning Calorimetry (DSC).

[0052] In another embodiment, Form A is characterized by having a DSC thermogram essentially the same as that shown in FIG.

[0053] In another embodiment, Form A has peaks at 1678, 1636, 1589, 1525, 1214, and 1196 cm -1 An infrared (IR) spectrum having a peak at cm -1 Value is ±4cm -1 It is characterized by having an IR spectrum in which

[0054] In another embodiment, Form A has the following molecular weights: 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 An IR spectrum having a peak at cm -1 Value is ±4cm -1 It is characterized by having an IR spectrum in which

[0055] In another embodiment, Form A is characterized by having an IR spectrum essentially the same as that shown in FIG.

[0056] In another embodiment, Form A has peaks of 5145, 4630, and 4423 cm -1 A near infrared (NIR) spectrum having a peak at cm -1 Value is ±4cm -1 It is characterized by having an NIR spectrum in which

[0057] In another embodiment, Form A has the molecular weights of 5908, 5796, 5145, 4875, 4630, 4423, and 4298 cm -1 A NIR spectrum having a peak at cm -1 Value is ±4cm -1It is characterized by having an NIR spectrum in which

[0058] In another embodiment, Form A is characterized by having a NIR spectrum essentially the same as that shown in FIG.

[0059] In another embodiment, the trophinetide polymorph, e.g., Form A, is characterized in that it contains about 1% to about 10% by weight, e.g., about 10% by weight, about 9% by weight, about 8% by weight, about 7% by weight, about 6% by weight, about 5% by weight, about 4% by weight, about 3% by weight, about 2% by weight, or about 1% by weight of any other physical form, e.g., crystalline or amorphous form of trophinetide or trophinetide hydrate.

[0060] In another embodiment, the trophinetide polymorph, e.g., Form A, is characterized in that it contains about 0.1% to about 1% by weight, e.g., 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 trophinetide or trophinetide hydrate.

[0061] In another embodiment, the trophinetide polymorph is characterized in that it does not contain a PXRD detectable amount of any other physical form of trophinetide or trophinetide hydrate. In another embodiment, the trophinetide polymorph is Form A.

[0062] In another embodiment, the trophinetide polymorph has an average particle size distribution of about 10 μm to about 500 μm, e.g., 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 trophinetide polymorph is Form A.

[0063] In another embodiment, the trophinetide polymorph has an average particle size distribution of about 1 μm to about 10 μm, e.g., 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 trophinetide polymorph is Form A.

[0064] In another embodiment, the trophinetide polymorph has an average particle size distribution that is about 1 μm or less, e.g., 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 trophinetide polymorph is Form A.

[0065] In another embodiment, the trophinetide polymorph is chemically stable during storage for 3 months at a temperature of about 25° C. and a relative humidity of about 60%. In another embodiment, the trophinetide polymorph is Form A.

[0066] In another embodiment, the trophinetide polymorph is chemically stable during storage for six months at a temperature of about 25° C. and a relative humidity of about 60%. In another embodiment, the trophinetide polymorph is Form A.

[0067] In another embodiment, the trophinetide polymorph is chemically stable during storage for greater than 12 months at a temperature of about 25° C. and a relative humidity of about 60%.

[0068] II. Pharmaceutical Compositions and Formulations In another embodiment, the present disclosure provides a pharmaceutical composition comprising a trophinetide polymorph and one or more pharma- ceutically acceptable excipients.

[0069] In another embodiment, the present disclosure provides a pharmaceutical formulation comprising the trophinetide polymorph in granular form, the granules optionally comprising one or more pharma- ceutically acceptable binders or fillers, or combinations thereof. The binders may be used in a total amount of about 1% to about 30% by weight of the granule, for example, in a total amount of about 5% to about 15% by weight of the granule, for example, 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% by weight of the granule. The fillers may be used in a total amount of about 5% to about 80% by weight of the granule, for example, about 10% to about 60% by weight of the granule, for example, 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% by weight of the granule.

[0070] In another embodiment, the binder is acacia, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, polyethylene glycol (PEG), povidone (polyvinylpyrrolidone, PVP), sucrose, or starch, or a combination thereof.

[0071] In another embodiment, the filler is microcrystalline cellulose.

[0072] In another embodiment, the present disclosure provides an aqueous pharmaceutical formulation comprising a trophinetide polymorph dissolved in water, and optionally one or more additional excipients.

[0073] In another embodiment, the water is purified water.

[0074] In another embodiment, about 1 g of trophinetide polymorph is dissolved in each 5 mL of water.

[0075] In another embodiment, the trophinetide polymorph is Form A.

[0076] In another embodiment, the pharmaceutical formulation contains 1% to 99% by weight of the trophinetide polymorph, for example, 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 depends on the effective dose of trophinetide, i.e., the dose required to elicit the desired level of therapeutic activity. In one embodiment, the pharmaceutical formulation comprises Form A dissolved in water.

[0077] III. Methods for making pharmaceutical formulations In another embodiment, the present disclosure provides a method for making an aqueous pharmaceutical formulation comprising trophinetide, comprising dissolving a trophinetide polymorph in water.

[0078] In another embodiment, the water is purified water.

[0079] In another embodiment, about 1 g of trophinetide polymorph is dissolved in each 5 mL of water.

[0080] In another embodiment, the trophinetide polymorph is Form A.

[0081] IV. Kit In another embodiment, the present disclosure provides a kit comprising a trophinetide polymorph packaged to facilitate its use for practicing the methods of the present disclosure.

[0082] In another embodiment, the kit comprises a trophinetide polymorph packaged in a container, such as a sealed bottle or vessel, with a label or package insert affixed to the container describing use of the trophinetide polymorph to practice the disclosed methods for treating a disease, disorder, or condition in a subject. In another embodiment, the trophinetide polymorph is packaged in a unit dosage form.

[0083] In another embodiment, the kit further comprises instructions for dissolving the trophinetide polymorph in water to provide an aqueous pharmaceutical formulation.

[0084] In another embodiment, the kit further comprises a package insert, e.g., instructions for administering the trophinetide polymorph or the aqueous pharmaceutical formulation to a subject having a disease, disorder, or condition. In another embodiment, the disease, disorder, or condition is traumatic brain injury. In another embodiment, the disease, disorder, or condition is a neurodevelopmental disorder. In another embodiment, the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0085] In another embodiment, the trophinetide polymorph is Form A.

[0086] V. METHODS OF TREATING DISEASES, DISORDERS, OR CONDITIONS In another embodiment, the present disclosure provides a method of treating a disease, disorder, or condition in a subject in need thereof, comprising administering to the subject an aqueous pharmaceutical formulation comprising a trophinetide polymorph dissolved in water. In another embodiment, the disease, disorder, or condition is traumatic brain injury. In another embodiment, the disease, disorder, or condition is a neurodevelopmental disorder. In another embodiment, the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0087] In another embodiment, the aqueous pharmaceutical formulation is a solution for oral administration.

[0088] In another embodiment, the water is purified water.

[0089] In another embodiment, about 1 g of trophinetide polymorph is dissolved in each 5 mL of water.

[0090] In another embodiment, the trophinetide polymorph is Form A.

[0091] VI. Methods for Making Crystalline Trofinetide or Trofinetide Hydrate In another embodiment, the present disclosure provides a method of making a trophinetide polymorph.

[0092] In another embodiment, the present disclosure provides a method of making Form A.

[0093] In another embodiment, the present disclosure provides a method of making Form A, comprising: i) adding ethanol to an aqueous solution of trophinetide 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 trophinetide is about 15% w / w.

[0094] In another embodiment, the method of making Form A further comprises adding Form A to seed the solution to obtain a slurry.

[0095] In another embodiment, the method of making Form A further comprises isolating the solid thus obtained, for example by filtration, to obtain a wet cake comprising Form A.

[0096] In another embodiment, the method of making Form A further comprises washing the wet cake comprising Form A with ethanol pre-chilled at about 0°C.

[0097] In another embodiment, the method of making Form A further comprises drying the wet cake comprising Form A under vacuum.

[0098] VII.Definition As used herein, the term "trophinetide" refers to a compound of formula I: [ka] Glycyl-L-2-methylprolyl-L-glutamic acid, with each stereocenter in the S configuration. The IUPAC name of trophinetide is (2S)-2-[[(2S)-1-(2-aminoacetyl)-2-methylpyrrolidine-2-carbonyl]amino]pentanedioic acid. Trofinetide is also referred to as "G-2-MePE", "H-Gly-MePro-Glu-OH", or "Gly-MePro-Glu-OH".

[0099] Trofinetide may form solvates, e.g., crystalline solids that incorporate water or methanol into the crystal lattice without chemical alteration of the trophinetide molecule. The term "trophinetide hydrate" refers to a compound of the formula: trophinetide·xH 2 O, where x is the number of H per mole of trophinetide. 2 0 molar ratio. Trofinetide hydrate need not contain a stoichiometric amount 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.

[0100] As used herein, the term "substantially pure" with respect to a trophinetide polymorph means that the crystalline material contains about 10% or less by weight, e.g., about 1% to about 10% by weight, e.g., about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, or about 1% by weight of any other crystalline or amorphous form of trophinetide or trophinetide hydrate. In another embodiment, the trophinetide polymorph is substantially pure Form A.

[0101] As used herein, the term "pure" with respect to a trophinetide polymorph means that the crystalline material contains about 1% or less by weight, e.g., about 0.1% to about 1% by weight, e.g., 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 by weight of any other crystalline or amorphous forms of trophinetide. In one embodiment, the crystalline trophinetide polymorph, e.g., Form A, does not contain a PXRD detectable amount of any other crystalline or amorphous forms of trophinetide or trophinetide hydrate. In another embodiment, the trophinetide polymorph is pure Form A.

[0102] As used herein, the term "amorphous" refers to a solid form of trophinetide or trophinetide hydrate that lacks the long range order characteristic of a crystal, ie, the solid is non-crystalline.

[0103] As used herein, the term "essentially the same" with respect to PXRD peak positions and / or relative intensities means that peak position and / or intensity variations are taken into account when comparing PXRD diffractograms.Similarly, the term "essentially the same" with respect to Raman or IR peak positions means that peak position variations are taken into account when comparing Raman or IR spectra.For example, PXRD peak positions may, for example, exhibit instrumental variations, for example, up to 0.2° 2θ, i.e. ±0.2° 2θ, and Raman and IR peak positions may, for example, exhibit instrumental variations, for example, up to 4 cm -1 , i.e. ±4cm -1 For example, in a PXRD diffractogram, the relative peak intensities may also show instrument-to-instrument variations due to the degree of crystallinity, orientation, sample surface preparation, and other factors known to those skilled in the art, and should be considered only as a qualitative measure.

[0104] As used herein, the term "atomization" refers to a process or method whereby a population of particles is reduced in size, typically to the micron scale.

[0105] As used herein, the term "micron" or "μm" means 1×10 -6 It refers to "micrometer", which is a unit of metre.

[0106] As used herein, the term "therapeutically effective amount" refers to an amount of trophinetide sufficient to treat one or more symptoms of a disease, condition, injury or disorder, or to prevent the progression of a disease, condition, injury or disorder, or to cause regression of a disease, condition, injury or disorder.

[0107] As used herein, terms such as "chemically stable" with respect to a trophinetide polymorph means that the trophinetide crystalline solid exhibits less than 0.5% chemical decomposition, e.g., less than 0.4%, less than 0.3%, less than 0.2%, less than 0.1%, or less than 0.05% chemical decomposition after storage for at least 3 months at a temperature of about 25° C. and a relative humidity of about 60%. In determining the amount of decomposition, methods known in the art, e.g., HPLC, can be used to measure the appearance of one or more chemical impurities and / or the disappearance of trophinetide.

[0108] The terms "a" and "an" refer to one or to more than one.

[0109] As used herein, the term "about" includes the stated numerical value plus or minus 10%. Thus, "about 10" means 9 to 11.

[0110] As used herein, "average particle size distribution" or "D 50 " is the diameter at which 50% by weight of the particles have a larger equivalent diameter and the other 50% by weight have a smaller equivalent diameter, as determined by laser diffraction, for example in a Malvern Master Sizer Microplus instrument or equivalent.

[0111] As used herein, the term "excipient" refers to any ingredient, other than the trophinetide polymorph, in or added to produce a pharmaceutical formulation suitable for administration to a subject, e.g., a solution for oral administration. An excipient is typically an inert substance, e.g., water, added to a composition to facilitate processing, handling, dissolution, administration, etc. of the trophinetide polymorph. Useful excipients include, but are not limited to, adjuvants, anti-adherents, binders, carriers, disintegrants, fillers, flavors, dyes, diluents, lubricants, glidants, preservatives, sorbents, solvents, surfactants, and sweeteners.

[0112] Conventional pharmaceutical excipients are well known to those skilled in the art. A wide variety of pharmaceutical excipients can be used in combination with the trophinetide polymorph, including water and others listed in Handbook of Pharmaceutical Excipients, Pharmaceutical Press 4th Ed. (2003), and Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st Ed. (2005). In one embodiment, the composition comprises Form A dissolved in water.

[0113] As used herein, the term "subject" refers to an animal, e.g., a human or a veterinary animal, e.g., a cow, sheep, pig, horse, dog, or cat. In one embodiment, the subject is a human.

[0114] As used herein, the term "container" therefore refers to any container and closure suitable for storing, shipping, distributing, and / or handling a pharmaceutical agent or excipient.

[0115] The term "package insert" means the information that accompanies a drug product that provides instructions on how to administer the product, along with safety and effectiveness data necessary to enable physicians, pharmacists, and patients to make informed decisions regarding the product's use. Package inserts are commonly considered the "label" of a drug product.

[0116] As used herein, the term "and / or" should be interpreted as a specific disclosure of each of the two specified features or components, with or without the other. Thus, the term "and / or" used in phrases such as "A and / or B" is intended to include "A and B," "A or B," "A" (single), and "B" (single). Similarly, the term "and / or" used in phrases such as "A, B, and / or C" is intended to encompass each of the following aspects: 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 (single); B (single); and C (single).

[0117] VII. Specific Embodiments The present disclosure provides the following specific embodiments.

[0118] Embodiment 1. Crystalline trophinetide xH 2 O, wherein x is from about 2 to about 4; (i) an X-ray powder diffraction pattern using Cu Kα radiation having peaks at 6.8, 11.5, 12.6, 13.8, and 16.8 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ; or (ii) a powder X-ray diffraction pattern using Cu Kα radiation with d spacings of 13.1, 7.7, 7.0, 6.4, and 5.3 Å; or (iii) 2989, 2934, 2883, 1685, 1637, 1459, and 930 cm -1 FT-Raman spectrum having a peak at cm -1 Value is ±4cm -1 FT-Raman spectrum, or (iv) 13, 24, 67, and 77 cm -1 A low frequency (LF) Raman spectrum with a peak at cm -1 Value is ±4cm -1 or the low-frequency (LF) Raman spectrum, (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; 13 C solid-state nuclear magnetic resonance spectrum, or (vi) from the furthest downfield peak, the delta to (i) the second furthest downfield peak is 1.8 ppm, (ii) the third furthest downfield peak is 2.2 ppm, (iii) the fourth furthest downfield peak is 2.5 ppm, (iv) the fifth furthest downfield peak is 2.7 ppm, (v) the sixth furthest downfield peak is 14.4 ppm, (vi) the seventh furthest downfield peak is 14.8 ppm, (vii) the eighth furthest downfield peak is 14.9 ppm, (viii) the ninth furthest downfield peak is 111.9 ppm, (ix) the tenth furthest downfield peak is 112.3 ppm, (x) the eleventh furthest downfield peak is 113.1 ppm, (y) the tenth furthest downfield peak is 114.1 ppm, (z) the tenth furthest downfield peak is 115.1 ppm, (z) the tenth furthest downfield peak is 116.1 ppm, (z) the tenth furthest downfield peak is 117.1 ppm, (z) the tenth furthest downfield peak is 118.1 ppm, (z) the tenth furthest downfield peak is 119.1 ppm, (z) the tenth furthest downfield peak is 120.1 ppm, (z) the tenth furthest downfield peak is 122.1 ppm, (z) the tenth furthest downfield peak is 124.1 ppm, (z) the tenth furthest downfield peak is 125.1 ppm, (z) the tenth furthest downfield peak is 126.1 ppm, (z) the tenth furthest downfield peak is 127.1 ppm, (z) the tenth furthest downfield peak is 128.1 ppm (xi) a delta to the twelfth furthest downfield peak is 121.1 ppm; (xi) a delta to the twelfth furthest downfield peak is 121.5 ppm; (xii) a delta to the thirteenth furthest downfield peak is 133.1 ppm; (xiii) a delta to the fourteenth furthest downfield peak is 139.4 ppm; (xiv) a delta to the fifteenth furthest downfield peak is 146.3 ppm; (xvi) a delta to the sixteenth furthest downfield peak is 154.6 ppm; (xvi) a delta to the seventeenth furthest downfield peak is 156.2 ppm; and / or (xvii) a delta from the furthest downfield peak to the furthest upfield peak is 158.6 ppm, or any combination thereof. 13 C solid-state nuclear magnetic resonance spectrum, or (vii) a melting point having an onset temperature of 71.71° C. and a peak temperature of 72.06° C. based on differential scanning calorimetry; or (viii) 1678, 1636, 1589, 1525, 1214, and 1196 cm -1 An infrared (IR) spectrum having a peak at cm -1 Value is ±4cm -1 IR spectrum, or (ix) 5145, 4630, and 4423 cm -1 A near infrared (NIR) spectrum having a peak at cm -1 Value is ±4cm -1 , or The crystalline trophinetide xH is characterized by having a combination thereof. 2 O.

[0119] Embodiment 2. Crystalline trophinetide xH 2 O, wherein x is from about 2 to about 4; (i) an X-ray powder diffraction pattern using Cu Kα radiation having peaks at 6.8, 11.5, 12.6, 13.8, and 16.8 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ; and / or (ii) a powder X-ray diffraction pattern using Cu Kα radiation with d spacings of 13.1, 7.7, 7.0, 6.4, and 5.3 Å; and / or (iii) 2989, 2934, 2883, 1685, 1637, 1459, and 930 cm -1 FT-Raman spectrum having a peak at cm -1 Value is ±4cm -1 and / or (iv) 13, 24, 67, and 77 cm -1 A low frequency (LF) Raman spectrum with a peak at cm -1 Value is ±4cm -1 and / or a low frequency (LF) Raman spectrum, (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; 13 C solid-state nuclear magnetic resonance spectroscopy, and / or (vi) from the furthest downfield peak, the delta to (i) the second furthest downfield peak is 1.8 ppm, (ii) the third furthest downfield peak is 2.2 ppm, (iii) the fourth furthest downfield peak is 2.5 ppm, (iv) the fifth furthest downfield peak is 2.7 ppm, (v) the sixth furthest downfield peak is 14.4 ppm, (vi) the seventh furthest downfield peak is 14.8 ppm, (vii) the eighth furthest downfield peak is 14.9 ppm, (viii) the ninth furthest downfield peak is 111.9 ppm, (ix) the tenth furthest downfield peak is 112.3 ppm, (x) the eleventh furthest downfield peak is 113.1 ppm, (y) the tenth furthest downfield peak is 114.1 ppm, (z) the tenth furthest downfield peak is 115.1 ppm, (z) the tenth furthest downfield peak is 116.1 ppm, (z) the tenth furthest downfield peak is 117.1 ppm, (z) the tenth furthest downfield peak is 118.1 ppm, (z) the tenth furthest downfield peak is 119.1 ppm, (z) the tenth furthest downfield peak is 120.1 ppm, (z) the tenth furthest downfield peak is 122.1 ppm, (z) the tenth furthest downfield peak is 124.1 ppm, (z) the tenth furthest downfield peak is 125.1 ppm, (z) the tenth furthest downfield peak is 126.1 ppm, (z) the tenth furthest downfield peak is 127.1 ppm, (z) the tenth furthest downfield peak is 128.1 ppm (xi) a delta to the twelfth furthest downfield peak is 121.1 ppm; (xi) a delta to the twelfth furthest downfield peak is 121.5 ppm; (xii) a delta to the thirteenth furthest downfield peak is 133.1 ppm; (xiii) a delta to the fourteenth furthest downfield peak is 139.4 ppm; (xiv) a delta to the fifteenth furthest downfield peak is 146.3 ppm; (xv) a delta to the sixteenth furthest downfield peak is 154.6 ppm; (xvi) a delta to the seventeenth furthest downfield peak is 156.2 ppm; and / or (xvii) a delta from the furthest downfield peak to the furthest upfield peak is 158.6 ppm, or any combination thereof. 13 C solid-state nuclear magnetic resonance spectroscopy, and / or (vii) a melting point based on differential scanning calorimetry having an onset temperature of 71.71° C. and a peak temperature of 72.06° C.; and / or (viii) 1678, 1636, 1589, 1525, 1214, and 1196 cm -1 An infrared (IR) spectrum having a peak at cm -1 Value is ±4cm -1 and / or (ix) 5145, 4630, and 4423 cm -1 A near infrared (NIR) spectrum having a peak at cm -1 Value is ±4cm -1 and / or The crystalline trophinetide xH is characterized by having a combination thereof. 2 O.

[0120] Embodiment 3. The crystalline trophinetide·xH according to embodiment 1 or 2, characterized in that it has an X-ray powder diffraction pattern using Cu Kα radiation with peaks at 6.8, 11.5, 12.6, 13.8, and 16.8 degrees 2θ, with 2θ values ​​of ±0.2 degrees 2θ. 2 O.

[0121] Embodiment 4. The crystalline trophinetide·xH according to any one of embodiments 1 to 3, characterized in that it has a powder X-ray diffraction pattern using Cu Kα radiation with d-spacings of 13.1, 7.7, 7.0, 6.4, and 5.3 Å. 2 O.

[0122] Embodiment 5. 2989, 2934, 2883, 1685, 1637, 1459, and 930 cm -1 FT-Raman spectrum having a peak at cm -1 Value is ±4cm -1 The crystalline trophinetide xH according to any one of embodiments 1 to 4, characterized in that it has an FT-Raman spectrum of 2 O.

[0123] 6.13, 24, 67, and 77 cm -1 A low frequency (LF) Raman spectrum with a peak at cm -1 Value is ±4cm -1 The crystalline trophinetide xH according to any one of embodiments 1 to 5, characterized in that it has a low frequency (LF) Raman spectrum of 2 O.

[0124] Embodiment 7. 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; 13 The crystalline trophinetide xH according to any one of embodiments 1 to 6, characterized by having a C solid-state nuclear magnetic resonance spectrum. 2 O.

[0125] Embodiment 8. From the furthest downfield peak, (i) the Δ to the seventh furthest downfield peak is 1.8 ppm, (ii) the Δ to the third furthest downfield peak is 2.2 ppm, (iii) the Δ to the fourth furthest downfield peak is 2.5 ppm, (iv) the Δ to the fifth furthest downfield peak is 2.7 ppm, (v) the Δ to the sixth furthest downfield peak is 14.4 ppm, (vi) the Δ to the seventh furthest downfield peak is 14.8 ppm, (vii) the Δ to the eighth furthest downfield peak is 14.9 ppm, (viii) the Δ to the ninth furthest downfield peak is 111.9 ppm, (ix) the Δ to the tenth furthest downfield peak is 112.3 ppm, (x) the 1st furthest downfield peak is 113.7 ppm, (x) the 2nd furthest downfield peak is 114.8 ppm, (x) the 3rd furthest downfield peak is 115.7 ppm, (x) the 4th furthest downfield peak is 116.7 ppm, (x) the 5th furthest downfield peak is 117.7 ppm, (x) the 6th furthest downfield peak is 118.7 ppm, (x) the 7th furthest downfield peak is 119.7 ppm, (x) the 8th furthest downfield peak is 120.7 ppm, (x) the 9th furthest downfield peak is 121.7 ppm, (x) the 10th furthest downfield peak is 122.7 ppm, (x) the 11th furthest downfield peak is 123.7 ppm, (x) the 12th furthest downfield peak is 124 (xi) a delta to the 1st furthest downfield peak is 121.1 ppm; (xi) a delta to the 12th furthest downfield peak is 121.5 ppm; (xii) a delta to the 13th furthest downfield peak is 133.1 ppm; (xiii) a delta to the 14th furthest downfield peak is 139.4 ppm; (xiv) a delta to the 15th furthest downfield peak is 146.3 ppm; (xvi) a delta to the 16th furthest downfield peak is 154.6 ppm; (xvi) a delta to the 17th furthest downfield peak is 156.2 ppm; and / or (xvii) a delta from the furthest downfield peak to the furthest upfield peak is 158.6 ppm, or any combination thereof. 13 The crystalline trophinetide xH according to any one of embodiments 1 to 7, characterized by having a C solid-state nuclear magnetic resonance spectrum. 2 O.

[0126] Embodiment 9. The crystalline trophinetide xH according to any one of embodiments 1 to 8, characterized in that it has a melting point with an onset temperature of 71.71°C and a peak temperature of 72.06°C based on differential scanning calorimetry. 2 O.

[0127] Embodiments 10.1678, 1636, 1589, 1525, 1214, and 1196 cm -1 An infrared (IR) spectrum having a peak at cm -1 Value is ±4cm -1 The crystalline trophinetide xH according to any one of embodiments 1 to 8, characterized in that it has an IR spectrum of 2 O.

[0128] Embodiments 11.5145, 4630, and 4423 cm -1 A near infrared (NIR) spectrum having a peak at cm -1 Value is ±4cm -1 The crystalline trophinetide xH according to any one of embodiments 1 to 10, characterized in that it has an NIR spectrum of 2 O.

[0129] Embodiment 12. The crystalline trophinetide xH according to any one of embodiments 1 to 11, having a mean particle size distribution of about 10 μm to about 500 μm. 2 O.

[0130] Embodiment 13. The crystalline trophinetide xH according to any one of embodiments 1 to 12, wherein x is about 2.5 to about 3.5. 2 O.

[0131] Embodiment 14. The crystalline trophinetide xH according to any one of embodiments 1 to 12, wherein x is about 2. 2 O.

[0132] Embodiment 15. The crystalline trophinetide xH according to any one of embodiments 1 to 12, wherein x is about 2.5. 2 O.

[0133] Embodiment 16. The crystalline trophinetide xH according to any one of embodiments 1 to 12, wherein x is about 3. 2 O.

[0134] Embodiment 17. The crystalline trophinetide xH according to any one of embodiments 1 to 12, wherein x is about 3.5. 2 O.

[0135] Embodiment 18. The crystalline trophinetide xH according to any one of embodiments 1 to 12, wherein x is about 4. 2 O.

[0136] Embodiment 19. Crystalline trophinetide xH according to any one of embodiments 1 to 18. 2 A pharmaceutical composition comprising O and a pharma- ceutically acceptable excipient.

[0137] Embodiment 20. The pharmaceutical composition according to embodiment 19 in granular form.

[0138] Embodiment 21. Crystalline trophinetide xH according to any one of embodiments 1 to 18 dissolved in water. 2 1. An aqueous pharmaceutical formulation comprising O.

[0139] About 1 gram of crystalline trophinetide xH 2 The aqueous pharmaceutical formulation according to embodiment 15, wherein O is dissolved in 5 mL of water each.

[0140] Embodiment 23. Crystalline trophinetide xH 2 23. A method of making the aqueous pharmaceutical formulation of embodiment 21 or 22, comprising mixing O with water.

[0141] Embodiment 24. Crystalline trophinetide xH according to any one of embodiments 1 to 18. 2 O and crystalline trophinetide xH in water 2 and instructions for dissolving O to provide an aqueous pharmaceutical formulation.

[0142] Embodiment 25. The kit of embodiment 24, further comprising instructions for administering the aqueous pharmaceutical formulation to a subject having a disease, disorder, or condition.

[0143] Embodiment 26 The kit of embodiment 25, wherein the disease, disorder or condition is traumatic brain injury.

[0144] Embodiment 27. The kit of embodiment 25, wherein the disease, disorder or condition is a neurodevelopmental disorder.

[0145] Embodiment 28 The kit of embodiment 27, wherein the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0146] Embodiment 29. A method for treating a disease, disorder or condition in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of embodiment 19 or 20, or the aqueous pharmaceutical formulation of embodiment 21 or 22.

[0147] Embodiment 30. The method of embodiment 29, wherein the disease, disorder or condition is traumatic brain injury.

[0148] Embodiment 31 The method of embodiment 29, wherein the disease, disorder or condition is a neurodevelopmental disorder.

[0149] Embodiment 32. The method of embodiment 31, wherein the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0150] Embodiment 33 The method of embodiment 29, wherein the disease, disorder or condition is Rett Syndrome.

[0151] Embodiment 34. The pharmaceutical composition of embodiment 19 or 20, or the aqueous pharmaceutical formulation of embodiment 21 or 22, for use in treating a disease, disorder or condition in a subject in need thereof.

[0152] Embodiment 35. The composition or formulation of embodiment 34, wherein the disease, disorder or condition is traumatic brain injury.

[0153] Embodiment 36. The composition or formulation of embodiment 34, wherein the disease, disorder or condition is a neurodevelopmental disorder.

[0154] Embodiment 37. The composition or formulation of embodiment 36, wherein the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0155] Embodiment 38. The composition or formulation of embodiment 34, wherein the disease, disorder or condition is Rett Syndrome.

[0156] Embodiment 39. Use of trophinetide of any one of embodiments 1 to 18, or the pharmaceutical composition of embodiment 19 or 20, or the aqueous pharmaceutical formulation of embodiment 21 or 22, in the manufacture of a medicament for a disease, disorder or condition in a subject in need of treatment thereof.

[0157] Embodiment 40. The use according to embodiment 39, wherein the disease, disorder or condition is traumatic brain injury.

[0158] Embodiment 41. The use according to embodiment 39, wherein the disease, disorder or condition is a neurodevelopmental disorder.

[0159] Embodiment 42. The use according to embodiment 41, wherein the neurodevelopmental disorder is Rett syndrome, fragile X syndrome, or autism spectrum disorder.

[0160] Embodiment 43. The use according to embodiment 39, wherein the disease, disorder or condition is Rett Syndrome.

[0161] Embodiment 44. Crystalline trophinetide xH according to any one of embodiments 1 to 18. 2 A method for preparing trophinetide·xHO comprising: i) adding ethanol to an aqueous solution of trophinetide at about 25° C.; ii) cooling the solution to about 0° C.; and iii) treating the solid thus obtained with crystalline trophinetide·xHO. 2 and isolating as O.

[0162] Embodiment 45. The method of embodiment 44, wherein the water:ethanol ratio is about 3:7 w / w. EXAMPLES

[0163] device Powder X-ray Diffraction (PXRD or XRPD) PXRD and XRPD are synonymous terms. The Rigaku Smart-Lab X-ray diffraction system was configured for reflection Bragg-Brentano geometry using a linear source X-ray beam. The X-ray source is a Cu Long Fine Focus tube (λ=1.54) operated at 40 kV and 44 mA. The source provides an incident beam profile at the sample that varies from a narrow line at high angles to a wide rectangle at low angles. Beam-tuning slits are used in the linear X-ray source to ensure that the maximum beam size is less than 10 mm both along the line and perpendicular to the line. The Bragg-Brentano geometry is a parafocusing geometry controlled by passive divergence and acceptance slits, with the sample itself acting as the focusing component of the optical system. The intrinsic resolution of the Bragg-Brentano geometry depends in part on the radius of the diffractometer used and the width of the acceptance slit. Typically, the Rigaku Smart-Lab is operated to give peak widths of 0.1° 2θ or less. The axial divergence of the X-ray beam is controlled by 5.0 degree Soller slits in both the incident and diffracted beam paths.

[0164] Powder samples were prepared in a low background Si holder using light manual pressure to keep the sample surface flat and level with the reference surface of the sample holder. Each sample was analyzed from 2 to 40°2θ using a continuous scan of 6°2θ per minute with an effective step size of 0.02°2θ.

[0165] Differential Scanning Calorimetry (DSC) DSC analysis was performed using a TA Instruments Q2500 Discovery Series instrument. Temperature calibration of the instrument was performed using indium. The DSC cell was maintained under a nitrogen purge of approximately 50 mL per minute during the analysis. Samples were placed in standard crimped aluminum pans and heated from approximately 25°C to 350°C at a rate of 10°C per minute.

[0166] Dynamic Vapor Sorption (DVS) Analysis DVS analysis was performed on a TA Instruments Q5000 Dynamic Vapor Sorption analyzer. The instrument was calibrated with standard weight and with sodium bromide standards for humidity. Samples were analyzed at 25°C with 10% relative humidity (RH) steps from 5 to 95% RH (adsorption cycle) and 95 to 5% RH (desorption cycle) with a maximum equilibration time of 60 minutes.

[0167] Infrared (IR) Spectroscopy Infrared spectra were obtained on a Nicolet 6700 FT-IR system using a Nicolet SMART iTR attenuated total reflectance device.

[0168] Near-infrared (NIR) spectroscopy Near infrared spectra were obtained on a Nicolet iS50 IR system. Trofinetide form A at approximately 1% w / w in dry KBr was placed in a DRIFT (Diffuse Reflectance Infrared Fourier Transform Spectroscopy) macrocup and analyzed in the 8000 and 400 wavenumber spectral ranges.

[0169] FT-Raman spectroscopy Fourier transform (FT) Raman spectra were obtained on a Nicolet model 6700 spectrometer interfaced to a Nexus Raman accessory module. The instrument is equipped with a Nd:YAG laser operating at 1024 nm, CaF 2 The NMR spectrometer consisted of a 3-inch NMR spectrometer, a 3-mm square NMR spectrometer, a 10 ...

[0170] 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 powders were analyzed in open air using a probe tip attachment. Spectra were obtained using a 36 cm -1 It was acquired using a static scan centered on and in the spectral range -575 to 575 cm -1 The data was collected over a 100% power, 1 second exposure time, and 32 accumulations. Wavelength calibration was verified using a sulfur reference standard. Data acquisition was performed using WiRE 3.4 software.

[0171] Karl Fischer (KF) analysis Karl Fischer analysis was performed using a Mettler-Toledo C20 Coulometric KF titrator. The instrument was calibrated using Hydranal water standards containing 1% water. The titrant was a solution of Hydranal in methanol.

[0172] 13 C solid-state nuclear magnetic resonance (NMR) spectroscopy Solid State 13 C cross-polarization magic angle spinning (CPMAS) experiments were performed on a Bruker Avance II 400 spectrometer equipped with a Doty probe (DSI-1630) 1H(19F) / X double resonance. Samples (109 mg) were loaded into a 4-mm 4 mm silicon nitride rotor closed with a Kel-F end cap for subsequent data acquisition. Adamantane was used as an external standard, set to the methylene signal of adamantane at 38.48 ppm on the TMS scale. The acquisition and processing parameters used are shown in the table below. [Table A] 9.4 Tesla ( 13 100MHz at C, 1At different magnetic fields, such as 400 MHz for H, the NMR spectrometer 13 It is possible to perform C CPMAS analysis. Parameters such as acquisition time, dwell time, recycle delay, spin speed, number of scans, etc. can be varied and optimized depending on the NMR spectrometer.

[0173] Single crystal structure determination X-ray diffraction analysis was carried out at a temperature of 150 K using Cu Kα radiation (λ=1.54 Å). 13 H 21 N 3 O 6 3(H 2 The monoclinic parameters and calculated volume of the α-acetylene complex (α-acetylene complex) are a = 18.8946(8) Å, b = 7.2849(3) Å, c = 27.8601(12) Å, β = 109.8540(16)°, and V = 3606.9(3) Å. 3 For Z=8 and formula weight 369.37, the calculated density is 1.360 g / cm 3 It is.

[0174] Example 1 Synthesis and characterization of Form A Form A was prepared according to the following method.

[0175] Method 1 19.9 mg of amorphous trophinetide was loaded into a mechanical grinding vessel with one metal ball. 7.6 mg of L-asparagine and 10 microliters of water were added to the grinding vessel. The vessel was sealed and milled in a Retsch mill at 100% power level for approximately 20 minutes. The resulting solid was removed and placed in a vacuum dryer to dry overnight. The resulting material was a mixture of Form A and crystalline L-aspartic acid by XRPD.

[0176] Method 2 18.6 mg of amorphous trophinetide was loaded into a mechanical grinding vessel with one metal ball. 8.0 mg of L-aspartic acid and 10 microliters of water were added to the grinding vessel. The vessel was sealed and ground in a Retsch mill for approximately 20 minutes. The resulting solid was allowed to dry in the open vessel before transferring the solid. The resulting material was a mixture of Form A and crystalline L-aspartic acid by XRPD. The same experiment was repeated using 20 mg of trophinetide amorphous form and 1 mg of L-aspartic acid. The resulting material was mostly Form A and traces of crystalline L-aspartic acid by XRPD. This material was used as a seed (3 mg) for another repeat experiment using only amorphous trophinetide (19.7 mg). The resulting solid was Form A by XRPD.

[0177] Method 3 A solution of trophinetide (300 mL, trophinetide amorphous form 32% w / w in water) was charged into absolute ethanol (1200 mL) at ambient temperature. 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. The resulting solid was Form A by XRPD.

[0178] Method 4 Form A was prepared according to Method 4 using the following steps.

[0179] At room temperature, 205.7 Kg of amorphous trophinetide (KF 5.0%, 195.4 Kg on a dry basis) is dissolved in 617 Kg of water in a reactor.

[0180] 2808 Kg of ethanol is added to the solution in the reactor at room temperature with vigorous mixing.

[0181] Cool the solution to 0-2 °C.

[0182] 1.0 Kg of trophinetide seeds are added to the reactor and the batch is allowed to rest for NLT 6 hours under slow stirring. Crystallization occurs.

[0183] The slurry in the reactor is filtered and separated using a filter dryer.

[0184] The wet cake is washed twice with 325 Kg of ethanol pre-chilled to 0-2°C.

[0185] The wet cake is vacuum dried until the bulk of the residual solvent has evaporated, then the jacket temperature of the filter dryer is raised to room temperature to complete the drying. 2 O (KF 13.5%, 147.0 Kg on a dry basis) was isolated (75% yield).

[0186] Method 5 Form A was prepared according to Method 5 using the following steps.

[0187] Ethanol (4479 Kg) is added to an aqueous solution of trophinetide (total weight 1272 Kg, 23.2 w / w% trophinetide, 294.5 Kg trophinetide, 977.5 Kg water) at room temperature with vigorous mixing.

[0188] Cool the solution to 0-2 °C.

[0189] 2.5Kg of crystalline trophinetide·xH 2 O seeds are added to the reactor and the batch is allowed to age for NLT 6 hours with slow stirring. Crystallization occurs.

[0190] The slurry in the reactor is filtered and separated using a filter dryer.

[0191] The wet cake is washed twice with 502 Kg of ethanol pre-chilled to 0-2°C.

[0192] The wet cake is vacuum dried until the bulk of the residual solvent has evaporated, then the jacket temperature of the filter dryer is raised to room temperature to complete the drying. 292.0 Kg of crystalline trophinetide xH 2 O (KF 14.3%, 250.2.0 Kg on a dry basis) was isolated (85% yield).

[0193] Method 6 Form A was prepared according to Method 6 using the following steps.

[0194] 58.1 g of crystalline trophinetide·xH 2 0 (KF 14.0%, 50.0 g on a dry basis) is dissolved in 100 g of water in reactor 1 at room temperature.

[0195] 233 g (296 mL) of ethanol is added to the solution in reactor 1 at room temperature with vigorous mixing. (Water / ethanol ratio is about 3 / 7 w / w, target solution concentration is about 15% w / w trophinetide).

[0196] 128 g of the solution in reactor 1 is transferred to reactor 2 (approximately 1 / 3 of the solution).

[0197] Cool reactor 2 to 0-2 °C.

[0198] 0.5 g of trophinetide seed material is slurried in 5 g of ethanol / water (95 / 5 w / w).

[0199] The trophinetide seed slurry is added to reactor 2 and the batch is allowed to age for NLT 2 hours with slow agitation. Nucleation occurs to produce a seed bed.

[0200] Transfer the remaining solution from reactor 1 to reactor 2 over NLT 2 h, maintaining reactor 2 at 0-2 °C and mixing well.

[0201] Charge 334 g (423 mL) of ethanol into reactor 1 and cool to 0-2 °C.

[0202] Transfer ethanol from reactor 1 to reactor 2 over NLT 2 h, maintaining reactor 2 at 0-2 °C and mixing well (final water / ethanol ratio is approximately 15 / 85 w / w).

[0203] The slurry in reactor 2 is aged at 0 to 2°C for at least 2 hours while being thoroughly mixed.

[0204] Prepare cake wash solution by mixing 25 g water and 475 g EtOH and cooling to 0-2 °C.

[0205] The slurry in reactor 2 is filtered, and the wet cake on the filter is washed at 0 to 2°C.

[0206] Vacuum dry the wet cake at 0-2 °C until the bulk of the residual solvent has evaporated, then warm the batch to room temperature to complete drying. 2 O (KF 13.5%, 46.0 g dry basis) was isolated (92% yield).

[0207] The structure of Form A was solved by single crystal X-ray diffraction. The structure showed three water molecules per trophinetide molecule. The asymmetric unit of Form A is shown in FIG. 8. Hydrogen atoms have been omitted for clarity. The structure of Form A shows that each of the three water molecules is hydrogen bonded to an oxygen or nitrogen atom of trophinetide. Each of the three water molecules is also hydrogen bonded to an adjacent water molecule.

[0208] The X-ray powder diffraction (XRPD) diffractogram of Form A is shown in Figure 1. The XRPD peak list (±0.2° 2θ) is set forth in Table 1. [Table 1]

[0209] The Raman spectrum of Form A is shown in Figure 2. Raman peak list (±4 cm -1 ) are listed in Table 2. [Table 2]

[0210] The low frequency (LF) Raman spectrum of Form A is shown in Figure 3. LF Raman peak list (±4 cm m-1 ) are listed in Table 3. [Table 3]

[0211] Form A 13 The C solid state nuclear magnetic resonance (ssNMR) spectrum is shown in Figure 4. The ssNMR peak list is given in Table 4. Selected peaks (Δppm) are given in Tables 5 and 6. [Table 4] [Table 5] [Table 6]

[0212] Form A melts with an onset temperature of 71.71° C. and a peak temperature of 72.06° C. based on DSC analysis. See FIG. 5.

[0213] The infrared (IR) spectrum of Form A is shown in Figure 6. IR Peak List (±4cm -1 ) are listed in Table 7. [Table 7]

[0214] The near infrared (NIR) spectrum of Form A is shown in Figure 7. NIR peak list (±4 cm -1 ) are listed in Table 8. [Table 8]

[0215] Form A is non-hygroscopic. Dynamic vapor sorption-desorption (DVS) analysis of Form A showed little water uptake when the material was exposed from 5% RH to 95% RH and little water loss when the material was exposed from 95% RH to 5% RH (Figure 9). The resulting material after DVS remained as trophinetide Form A by XRPD (Figure 10).

[0216] Form A exhibits a weight loss of 12-14% between approximately 50-120 °C. This is likely due to water loss and is consistent with the 12-14% water content measured by Karl Fischer analysis. An exemplary TGA scan is shown in Figure 12. DSC data for Form A consistently shows a sharp endotherm at approximately 70-72 °C (peak temperature) corresponding to melting.

[0217] Form A is a trihydrate based on the single crystal X-ray structure which showed three water molecules per molecule of trophinetide (Figure 8). However, form A has been produced with a water content of about 12% to about 14%. This suggests that while some of the water is essential to the crystal lattice, at least one of the water molecules can be loosely bound and removed without altering the crystal lattice. For this reason, form A is a trihydrate of trophinetide·xH 2 O, where x is from about 2 to about 4.

[0218] Example 2 Stability of Form A Form A is stable under a wide range of humidity conditions. Form A exposed to 33%, 59%, 75%, and 97% RH for 2 days showed 2-4 molar equivalents of water. Under extremely dry conditions (open container exposed to 0% RH for 2 days), Form A lost water and became disordered. The Form A signal was still visible in the XRPD pattern, but the crystalline signal was widespread and the XRPD pattern showed some amorphous halos at the baseline, indicating the formation of disorder and amorphous material (Figure 11).

[0219] The long-term (6 months) chemical stability of Form A and amorphous trophinetide was tested under the same conditions of 25±2° C. / 60±5% relative humidity (RH). Form A is surprisingly more stable than amorphous trophinetide under these conditions (Table 9). [Table 9]

[0220] The analytical methods used for the impurity assays are described in Table 10. [Table 10]

[0221] It is to be understood that the foregoing described embodiments and examples are not intended to limit the scope of the present disclosure in any way, and that the claims presented herein are intended to encompass all embodiments and examples, whether or not expressly presented herein.

[0222] All patents and publications cited herein are incorporated by reference in their entirety.

Claims

Claim 1 Crystalline trophinetide·xH ２ O (wherein x is from about 2 to about 4), and (i) A powder X-ray diffraction pattern having peaks in the range of 6.6 to 6.8 degrees 2θ, in the range of 11.3 to 11.6 degrees 2θ, in the range of 12.5 to 12.7 degrees 2θ, and in the range of 13.6 to 13.8 degrees 2θ, using Cu Kα radiation, wherein the 2θ value is ±0.2 degrees 2θ, or (ii) A powder X-ray diffraction pattern having d-spacings of 13.1, 7.7, 7.0, 6.4, and 5.3 Å, using Cu Kα radiation, or (iii) An FT-Raman spectrum having peaks at 2989, 2934, 2883, 1685, 1637, 1459, and 930 cm-1, wherein the cm-1 value is ±4 cm-1, or (iv) A low-frequency (LF) Raman spectrum having peaks at 13, 24, 67, and 77 cm-1, wherein the cm-1 value is ±4 cm-1, or (v) A 13C solid nuclear magnetic resonance spectrum 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, wherein the ppm value is ±3 ppm, or (vi) The Δ from the farthest downfield peak to (i) the second farthest downfield peak is 1.8 ppm, (ii) the third farthest downfield peak is 2.2 ppm, (iii) the fourth farthest downfield peak is 2.5 ppm, (iv) the fifth farthest downfield peak is 2.7 ppm, (v) the sixth farthest downfield peak is 14.4 ppm, (vi) the seventh farthest downfield peak is 14.8 ppm, (vii) the eighth farthest downfield peak is 14.9 ppm, (viii) the ninth farthest downfield peak is 111.9 ppm, (ix) the tenth farthest downfield peak is 112.3 ppm, (x) the eleventh farthest downfield peak is 121.1 ppm, (xi) the twelfth farthest downfield peak is 121.5 ppm, (xii) the thirteenth farthest downfield peak is 133.1 ppm, (xiii) the fourteenth farthest downfield peak is 139.4 ppm, (xiv) the fifteenth farthest downfield peak is 146.3 ppm, (xv) the sixteenth farthest downfield peak is 154.6 ppm, (xvi) the seventeenth farthest downfield peak is 156.2 ppm, and / or (xvii) the Δ from the farthest downfield peak to the farthest upfield peak is 158.6 ppm, or any combination thereof, a 13C solid nuclear magnetic resonance spectrum having 18 peaks, or (vii) A melting point having an onset temperature of 71.71 °C and a peak temperature of 72.06 °C based on differential scanning calorimetry, or (viii) An infrared (IR) spectrum having peaks at 1678, 1636, 1589, 1525, 1214, and 1196 cm-1, wherein the cm-1 values are ±4 cm-1, an IR spectrum, or (ix) A near-infrared (NIR) spectrum having peaks at 5145, 4630, and 4423 cm-1, wherein the cm-1 values are ±4 cm-1, a NIR spectrum, or Crystalline trophynetide xH, characterized by having those combinations ２ O.

2. A powder X-ray diffraction pattern having peaks at 2θ of 6.7 or 6.8, 11.4 or 11.5, 12.6, and 13.7 or 13.8 degrees using Cu Kα radiation, wherein the 2θ value is ±0.2 degrees 2θ, the crystalline trophinetide.xH according to claim 1, characterized by having such a powder X-ray diffraction pattern ２ O.

3. A powder X-ray diffraction pattern having peaks at 2θ of 6.7, 11.4, 12.6, 13.7, 22.3, 23.6, 25.3 and 28.1 degrees 2θ using Cu Kα radiation, wherein the 2θ value is ±0.2 degrees 2θ, the crystalline trophinetide·xH according to claim 2, characterized in that it has a powder X-ray diffraction pattern ２ O.

4. The crystalline trophinetide.xH according to claim 1, characterized by having a powder X-ray diffraction pattern with d-spacings of 13.1, 7.7, 7.0, 6.4, and 5.3 Å using Cu Kα radiation ２ O.

5. FT-Raman spectra having peaks at 2989, 2934, 2883, 1685, 1637, 1459, and 930 cm －１ wherein the cm －１ value is ±4 cm －１ characterized by having an FT-Raman spectrum, of the crystalline trophinetide·xH ２ O according to claim 1. Claim 6 13, 24, 67, and 77 cm －１ having a peak in a low-frequency (LF) Raman spectrum, said cm －１ value being ±4 cm －１ characterized by having a low-frequency (LF) Raman spectrum, the crystalline trophinetide·xH ２ O according to claim 1. Claim 7 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 A 13C solid nuclear magnetic resonance spectrum, wherein the ppm value is ±3 ppm １３ The crystalline trophinetide·xH2O according to claim 1, characterized by having a 13C solid nuclear magnetic resonance spectrum ２ O. Claim 8 The crystalline trophinetide xH ２ O having a melting point based on differential scanning calorimetry and having an onset temperature of 71.71 °C and a peak temperature of 72.06 °C, as claimed in claim 1. Claim 9 An infrared (IR) spectrum having peaks at 1678, 1636, 1589, 1525, 1214, and 1196 cm －１ wherein the cm －１ value is ±4 cm －１ characterized by having an IR spectrum, of the crystalline trophinetide·xH ２ O according to claim 1. Claim 10 5145, 4630, and 4423 cm －１ with peaks in the near-infrared (NIR) spectrum, wherein the cm －１ value is ±4 cm －１ characterized by having an NIR spectrum, the crystalline trophinetide xH ２ O according to claim 1. Claim 11 Crystalline trophinetide xH₂O according to claim 1, wherein x is from about 2 to about 3.5 ２ O. Claim 12 Crystalline trophinetide·xH₂O according to claim 1, wherein x is about 3 ２ O. Claim 13 Crystalline trofinetide·xH ２ O and a pharmaceutically acceptable excipient, a pharmaceutical composition. Claim 14 An aqueous pharmaceutical formulation comprising the crystalline trophinetide·xH ２ O dissolved in water as claimed in claim 1. Claim 15 The crystalline trophinetide·xH ２ A method for producing the aqueous pharmaceutical preparation according to claim 14, which comprises mixing the O and water. Claim 16 Crystalline trophinetide·xH described in claim 1 ２ O and instructions for providing an aqueous pharmaceutical formulation by dissolving crystalline trophinetide·xH ２ O in water, a kit comprising the same. Claim 17 A pharmaceutical composition according to claim 13 for treating a disease, disorder or medical condition in a subject in need thereof, wherein the disease, disorder or medical condition is traumatic brain injury, neurodevelopmental disorder, Rett syndrome, fragile X syndrome, or autism spectrum disorder. Claim 18 The pharmaceutical composition according to claim 17, wherein the disease, disorder or medical condition is Rett syndrome. Claim 19 A method for preparing crystalline trophinetide·xH ２ O, comprising: i) adding ethanol to an aqueous solution of trophinetide at about 25°C; ii) cooling the solution to about 0°C; and iii) isolating the solid thus obtained to obtain crystalline trophinetide·xH ２ O, the method comprising the above steps.

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