Polymorphs of trofinetide

Abstract

The present invention discloses polymorphic forms of Trofinetide and preparation thereof. The invention also relates to pharmaceutical composition comprising novel polymorphs of Trofinetide and their use in treatment of neurological disorders.

Description

[0001] “POLYMORPHS OF TROFINETIDE”

[0002] Technical field of the Invention

[0003] The present invention relates to polymorphs of Trofinetide and processes for preparation thereof.

[0004] Background of Invention

[0005] Trofinetide is chemically known as (2S)-2-{[(2S)-l-(2-aminoacetyl)-2-methylpyrrolidine-2-carbonyl] amino}pentanedioic acid and represented as shown below in Structure-I.

[0006]

[0007] Structure-I

[0008] Trofinetide is a novel synthetic analog of glypromate, also known as glycine-proline-glutamate (GPE), a naturally occurring protein in the brain and the N-terminal tripeptide of insulin-like growth factor I. GPE exhibits neuroprotective properties. It protects neurons from glutamate-mediated excitotoxicity and oxidative stress at nanomolar concentrations. Trofinetide is a drug used to treat Rett syndrome in children and adults which is an X-linked neurodevelopmental disorder characterized by a range of cognitive, motor, and autonomic symptoms.

[0009] US7041314 discloses Trofinetide, methods of making Trofinetide, and methods of using Trofinetide to treat a disease, disorder, or condition, e.g., neural degeneration caused by hypoxia-ischemia or toxic injury. US 11827600 discloses crystalline Form A of Trofinetide hydrate, pharmaceutical compositions comprising crystalline Form A of Trofinetide hydrate, methods of making crystalline Form A of Trofinetide hydrate, and methods of treating a disease, condition, or disorder in a subject comprising administering a composition comprising crystalline Form A of Trofinetide hydrate.

[0010] US9212204 relates to a method for treating a human suffering from a symptom of Rett Syndrome.

[0011] OBJECT OF THE INVENTION

[0012] It is an object of the present invention to provide novel polymorphs of Trofinetide.

[0013] It is another object of the present invention to provide processes for the preparation of polymorphs of Trofinetide.

[0014] It is further object of the invention to provide a pharmaceutical composition comprising novel polymorphs of Trofinetide along with pharmaceutically acceptable carrier, diluent or excipients.

[0015] It is another object of the invention to use pharmaceutical composition as defined above for the treatment of various neurological disorders.

[0016] SUMMARY OF THE INVENTION

[0017] In an aspect, the present invention provides Trofinetide Form Cl.

[0018] In another aspect, the present invention provides Trofinetide Form Cl characterized by X-ray powder diffraction pattern (XRPD) having peaks as disclosed in Fig 1; Differential scanning calorimetry (DSC) with endothermic peaks as disclosed in Fig 2; Infrared (IR) spectroscopy having peaks as disclosed in Fig 3; and Fourier transform (FT)-Raman spectroscopy having peaks as disclosed in Fig 4. In another aspect, the present invention provides Trofinetide Form C2.

[0019] In another aspect, the present invention provides Trofinetide Form C2 characterized by X-ray powder diffraction pattern (XRPD) having peaks as disclosed in Fig 5.

[0020] In another aspect, the present invention provides processes for preparation of Trofinetide Form Cl and Form C2.

[0021] In an aspect, the present invention provides a process for preparing Trofinetide Form Cl comprising;

[0022] a. Dissolving Trofinetide base in a suitable solvent or mixture of solvents and stirring;

[0023] b. Optionally adding Trofinetide Form Cl seed dissolved in a solvent to the solution of step (a) and stirring;

[0024] c. Filtering and drying at a temperature ranging between 20°C to 50°C to obtain the desired product.

[0025] In another aspect, the present invention provides a process for preparing Trofinetide Form C2 comprising;

[0026] a. Charging a suitable solvent or solvent mixture in a reaction vessel; b. Seeding with Trofinetide Form Cl;

[0027] c. Adding Trofinetide to the above reaction mixture and stirring;

[0028] d. Heating the mixture at a temperature in the range of 40-45°C and maintaining at said temperature for a suitable period of time;

[0029] e. Cooling, filtering and drying to obtain the product.

[0030] In further aspect of the invention, there is provided a pharmaceutical composition comprising novel polymorphs of Trofinetide Form Cl and Form C2 and pharmaceutically acceptable carrier, diluent or excipients.

[0031] In yet another aspect of the invention there is provided the use of pharmaceutical composition comprising Trofinetide Form Cl and Form C2 for the treatment of neurodevelopmental disorders. DESCRIPTION OF FIGURES

[0032] Figure 1: illustrates X-ray powder diffraction spectrum (XRPD) of Trofinetide Form Cl.

[0033] Figure 2: illustrates Differential scanning calorimetry (DSC) of Trofinetide Form Cl

[0034] Figure 3: illustrates Infrared (IR) spectrum of Trofinetide Form Cl

[0035] Figure 4: illustrates Fourier transform (FT)-Raman spectrum of Trofinetide Form Cl

[0036] Figure 5: illustrates X-ray powder diffraction spectrum (XRPD) of Trofinetide Form C2.

[0037] DETAILED DESCRIPTION OF THE INVENTION

[0038] The invention will now be described in detail in its preferred and optional embodiments however should not be construed to limit the scope of the invention. Polymorphs are crystalline modifications or crystalline forms of a substance that can exist in more than one crystalline structure Therefore, the word "polymorph" comes from the Greek words poly (many) and morphe (form). A single molecule may have different polymorphs having distinct crystal structures and distinct physical properties such as X-ray powder diffraction, Thermogravimetric analysis (TGA), Differential scanning calorimetry (DSC), Infrared absorption (IR) etc. As used herein, the term " XRPD" refers to X-ray powder diffraction, “DSC” refers to Differential scanning calorimetry, “IR” refers to Infrared spectrum and FT-Raman refers to Fourier transform- Raman spectrum.

[0039] In an embodiment, the present invention relates to Trofinetide Form Cl characterized by having a XRPD pattern as shown in Figure 1.

[0040] In an embodiment, the present invention relates to o Trofinetide Form Cl characterized by XRPD pattern having diffraction peaks at 4.82, 6.82, 10.80, 12.35, 20.00 ± 0.2°20.

[0041] In another embodiment, Trofinetide Form Cl is further characterized having diffraction peaks at 9.68, 15.32, 18.47, 19.43, 24.33 ± 0.2°20. In another embodiment, Trofinetide Form Cl of the present invention is characterized by Differential Scanning Calorimetry (DSC) as having a melting point with a peak temperature of 108.15° C and 157.38° C and as shown in Fig 2.

[0042] In yet another embodiment, the present invention relates to Trofinetide Form Cl characterized by having a IR spectrum as shown in Figure 3.

[0043] Accordingly, the present invention relates to Trofinetide Form Cl characterized by IR spectrum as having peaks at 3293, 2938, 2054, 2017, 1986, 1633, 1527, 1454, 1432, 1410, 1347, 1248, 1216, 1194, 1147, 1022, 962, 921, 895, 838, 812, 770, 641, 614, 550 and 519 cm'1.

[0044] In another embodiment, the present invention relates to Trofinetide Form Cl characterized by having a FT-Raman spectrum as shown in Figure 4. Accordingly, Trofinetide Form Cl is characterized by FT-Raman spectrum having peaks at 2982, 2939, 2883, 1661, 1454, 1413, 1024, 932, 768, 709, 556 and 80 cm'1.

[0045] In another embodiment, the present invention relates to Trofinetide Form C2 characterized by having a XRPD pattern as shown in Figure 5.

[0046] In an embodiment, the present invention discloses Trofinetide Form C2 having XRPD pattern with characteristics diffraction peaks at 4.79, 6.84, 10.79, 18.18, 18.94 ± 0.2°20.

[0047] In an embodiment, the XRPD pattern of Trofinetide Form C2 having further diffraction peaks at 9.79, 12.45, 15.78, 20.12 ± 0.2°20.

[0048] In an embodiment, the present invention relates to a process for preparation of Trofinetide Form Cl and Trofinetide Form C2.

[0049] In another embodiment, the present invention relates to a process for preparation of Trofinetide Form Cl comprising; a. Dissolving Trofinetide base in a suitable solvent or mixture of solvents and stirring;

[0050] b. Optionally adding Trofinetide Form Cl seed dissolved in a solvent to the solution of step (a) and stirring;

[0051] c. Filtering and drying to obtain the desired product.

[0052] Accordingly, to the solvent or solvent mixture comprising of C1-C4 alcohols, water was added to Trofinetide base and the mixture was stirred, filtered and sucked dried under nitrogen gas. The resultant mixture was further dried under vacuum to obtain Trofinetide Form Cl.

[0053] In the optional embodiment, the process for preparing Trofinetide Form Cl comprises seeding Trofinetide dissolved in a solvent with the solution of Trofinetide Form Cl seed and stirring the mixture and optionally heating. Adding suitable solvent to the reaction mass, cooling, filtering and drying to obtain Trofinetide Form Cl.

[0054] In an embodiment, the suitable solvent used to dissolve Trofinetide may be selected from, but not limited to C1-C4 alcohol, ketone, acid, ester, nitrile, water or mixtures thereof. Preferably, the C1-C4 alcohol is selected from methanol, ethanol, isopropanol, n-propanol, isobutanol, n-butanol, and t-butanol. Most preferably C1-C4 alcohol is selected from methanol, ethanol, n-propanol and isopropanol.

[0055] In an embodiment, the process is carried out at a temperature in the range of 15°C to 35°C, preferably in the range of 20° C to 30° C, for a period of 25 hours to 50 hours, preferably for 30 hours to 45 hours.

[0056] In an embodiment, Trofinetide polymorph obtained after filtration may be dried under vacuum at a temperature in the range of about 20° C to 50°C, more preferably about 30°C to 40°C for a period of 1 hour to 10 hours, preferably, for 5 to 6 hours.

[0057] In another embodiment, the present invention relates to a process for preparation of Trofinetide polymorph C2 comprising;

[0058] a. Charging a suitable solvent or solvent mixture in a reaction vessel; b. Seeding with Trofinetide Form Cl;; c. Adding Trofinetide to the above reaction mixture and stirring;

[0059] d. Heating the mixture at a temperature in the range of 40-45°C and maintaining at said temperature for a suitable period of time;

[0060] e. Cooling filtering and drying to obtain the product.

[0061] In an aspect, the solvent used for preparation of Form C2 is selected from C1-C4 alcohol, acid, ketone, ester, nitrile, water or mixtures thereof.

[0062] According to the present process for preparation of Trofinetide Form C2, to the mixture of the solvents selected from C1-C4 alcohol and water was added to Trofinetide Form Cl seeds and the mixture was stirred for about 5-10 minutes. This was followed by addition of Trofinetide base and stirred for a period of 5-10 hours at ambient temperature. The mixture was then heated to a temperature in the range of 35-45 hours and maintained at said temperature for 15-30 hours. The contents were cooled to ambient temperature and the mixture was stirred for 12 to 15 hours. The reaction mass was filtered and dried to obtain the product.

[0063] In an embodiment, Trofinetide polymorphs Cl and C2 may be anhydrous or in hydrated form.

[0064] In an embodiment, the present invention relates to a pharmaceutical composition comprising polymorphic Trofinetide Form Cl or Trofinetide Form Cl prepared by the present process together with pharmaceutically acceptable excipients.

[0065] In yet another embodiment, the present invention relates to use of the pharmaceutical composition comprising polymorphic Trofinetide Form Cl or Trofinetide Form C2 in treating or preventing neurological disorder or treatment of stroke or heart related disease.

[0066] The invention will now be further described by the following examples, which are illustrative rather than limiting.

[0067] Examples:

[0068] Example 1: Process for preparation of Trofinetide Form Cl

[0069] A mixture of 4.8ml Ethanol and 0.2 ml water was charged in a clean round bottom flask (RBF). 0.5 g of Trofinetide base was added into this RBF and stirred for 35 to 40 hours. The resulted reaction mass was filtered through pressure nutsche filter (PNF) and sucked dried under nitrogen gas for 1 hour. The resultant material was dried under vacuum for 5 to 6 hours at about 35 °C to obtain Trofinetide Form Cl.

[0070] Yield: 0.25 g

[0071] Example 2: Process for preparation of Trofinetide Form Cl

[0072] 0.5 g of Trofinetide Form Cl seed and 20 ml of ethanol were charged in a clean round bottom flask (RBF). This was followed by addition of 5.0 g of Trofinetide and 30 ml of Ethanol were added into this RBF and the contents were stirred for 15 to 20 hours. 2 ml of water was added into the same reaction mass and maintained for about 6 to 8 hours. The reaction mass was then filtered through pressure nutsche filter (PNF) and sucked dried under nitrogen for about 1 hour. The resultant mixture was dried for 5 to 6 hours at about 35°C to obtain Trofinetide Form Cl.

[0073] Yield: 4 g

[0074] Example 3: Process for preparation of Trofinetide Form Cl

[0075] 300 g of Trofinetide was added in a lot wise manner into a mixture of 2490 ml ethanol and 120 ml water at room temperature and seeded with 15 g of Trofinetide Form Cl seeds and maintained at said temperature for 10 hrs.. Heated the reaction mass to 25-35°C for 1 hr, then added 4 V of ethanol and maintained the reaction mixture at 25-35°C for 3 hrs. Cooled the reaction mass to room temperature and maintained for 3 hrs. Filtered the reaction mass under nitrogen atmosphere in centrifuge followed by drying at 25-30°C in vacuum tray dryer for 24 hours. Comilled the reaction mass which was suspended in 8V Ethanol and 0.2 V of water at room temperature. Further heated to 25-35°C and maintained at said temperature for 1 hour. 2 V ethanol was further added at 25-35°C and the reaction mixture was maintained for 2 hours. Cooled the reaction mass to 25°C and stirred for 3 hours followed by filtration under nitrogen atmosphere in centrifuge and drying at 25-30°C in vacuum tray dryer for 24 hours. Co-milled the material and dried at 25-35°C for 48 hours in air tray dryer to obtain Trofinetide Form Cl. Yield: 198 g Example 4: Process for preparation of Trofinetide Form C2

[0076] A mixture of 7 ml of Isopropyl alcohol and 0.1 ml of water was charged in a clean round bottom flask (RBF). 50 mg of Form Cl seed was added into the above RBF and stirred for 5 min. 0.5 g of Trofinetide material was added into the above RBF and stirred for 6 to 8 hrs at 25°C. The contents were heated to 40°C and maintained the reaction mass for 20 hrs at 40°C. Cooled contents to 25°C and stirred for 12 to 15 hrs. The reaction mass was filtered through pressure nutsche filter (PNF) and sucked dried under nitrogen gas for 1 hr. The resultant material was dried under vacuum for 5 to 6 hrs at about 35°C to obtain Trofinetide Form C2.

[0077] Yield: 0.25g

[0078] Although the invention has been described in detail in the foregoing for the purpose of illustration, it is to be understood that such detail is solely for that purpose and that variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention except as it may be limited by the claims.

Claims

We claim:

1. Trofinetide Form Cl characterized by an X-ray powder diffraction pattern comprising diffraction peaks at 4.82±0.2°, 6.82±0.2°, 10.80±0.2°, 12.35±0.2° and 20.00 ±0.2° 20.

2. The Trofinetide Form Cl as claimed in Claim 1, characterized by an X-ray powder diffraction pattern comprising diffraction peaks at 9.68, 15.32, 18.47, 19.43, 24.33 ± 0.2°20.

3. The Trofinetide Form Cl as claimed in claim 1 or 2, characterized by a differential scanning calorimetry (DSC) curve comprising a melting point with an peak temperature of 108.15°±5°C and 157.38° ±5°C.

4. The Trofinetide Form Cl as claimed in claim 1 or 2, characterized by IR spectrum comprising peaks at 3293, 2938, 2054, 2017, 1986, 1633, 1527, 1454, 1432, 1410, 1347, 1248, 1216, 1194, 1147, 1022, 962, 921, 895, 838, 812, 770, 641, 614, 550 and 519 cm'1.

5. The Trofinetide Form Cl as claimed in claim 1 or 2, characterized by FT- Raman spectrum comprising peaks at 2982, 2939, 2883, 1661, 1454, 1413, 1024, 932, 768, 709, 556 and 80 cm'1.

6. Trofinetide Form C2, characterized by an X-ray powder diffraction pattern comprising diffraction peaks at 4.79±0.2°, 6.84±0.2°, 9.79±0.2°, 10.79±0.2°, 12.45±0.2°, 15.78±0.2°, 18.18±0.2°, 18.94±0.2° and 20.12 ±0.2° 20.

7. A process for preparation of Trofinetide Form Cl as claimed in claim 1 or 2 comprising;a. Dissolving Trofinetide base in a solvent or mixture of solvents and stirring;b. Optionally adding Trofinetide Form Cl seed dissolved in a solvent to the solution of step (a) and stirring;c. Filtering and drying to obtain the desired product.

8. The process as claimed in claim 7, wherein the solvent is selected from C1-C4 alcohol, ketone, acid, ester, nitrile, water or mixtures thereof.

9. The process as claimed in claim 7, wherein the process is carried out at a temperature in the range of 15°C to 35°C, preferably in the range of 20° C to 30° C.

10. The process for preparation of Trofinetide polymorph C2 as claimed in claim 6, comprising;a. Charging a solvent or solvent mixture in a reaction vessel;b. Seeding with Trofinetide Form Cl;;c. Adding Trofinetide to the above reaction mixture and stirring;d. Heating the mixture at a temperature in the range of 40-45°C and maintaining at said temperature for a suitable period of time;e. Cooling filtering and drying to obtain the product.

11. A pharmaceutical composition comprising Trofinetide Form Cl or Form C2 as claimed in any one of the preceding claims 1 to 10 together with acceptable excipients.

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