Polymorphic form of sepiapterin
Disclosed is a crystalline form of sepiapterin, a method of preparing the crystalline form, pharmaceutical compositions containing the crystalline form, and a method for treating patients with a disease associated with low intracellular BH4 levels or with dysfunction of various BH4 dependent metabolic pathways, which involves administering to the patient an effective amount of the crystalline form.
1. A method of preparing crystalline Form A of sepiapterin free base having peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 12.4°±0.5, 26.2°±0.5, and 28.9°±0.5 as measured by X-ray diffractometry by irradiation with Cu Kα X-rays, comprising:
(i) combining sepiapterin free base and hydrochloric acid;
(ii) isolating the hydrochloride salt of sepiapterin formed in step (i); and
(iii) neutralizing the hydrochloride salt of sepiapterin obtained in step (ii) with a base to obtain crystalline Form A of sepiapterin free base.
2. A method of preparing crystalline Form A of sepiapterin free base having peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 12.4°±0.5, 26.2°±0.5, and 28.9°±0.5 as measured by X-ray diffractometry by irradiation with Cu Kα X-rays, comprising dissolving sepiapterin free base in dimethyl acetamide, adding to the solution acetone, ethyl acetate, or THF, and isolating the solids to obtain the crystalline Form A of sepiapterin free base.
3. A method of preparing crystalline Form A of sepiapterin free base having peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 12.4°±0.5, 26.2°±0.5, and 28.9°±0.5 as measured by X-ray diffractometry by irradiation with Cu Kα X-rays, comprising dissolving sepiapterin free base in dimethyl sulfoxide, adding to the solution isopropyl alcohol, and cooling the solution to obtain the crystalline Form A of sepiapterin free base.
4. A method of preparing crystalline Form A of sepiapterin free base having peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 12.4°±0.5, 26.2°±0.5, and 28.9°±0.5 as measured by X-ray diffractometry by irradiation with Cu Kα X-rays, comprising preparing a slurry of sepiapterin free base in methyl tert-butyl ether, n-heptane, toluene, a mixture of chloroform and n-heptane, or a mixture of acetone and methyl tert-butyl ether, stirring the resulting suspension, and isolating the solids to obtain the crystalline Form A of sepiapterin free base.
5. A method of preparing crystalline Form A of sepiapterin free base having peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 12.4°±0.5, 26.2°±0.5, and 28.9°±0.5 as measured by X-ray diffractometry by irradiation with Cu Kα X-rays, comprising exposing sepiapterin free base to a vapor of water, methyl t-butyl ether, n-heptane, or toluene, and obtaining the crystalline Form A of sepiapterin free base.
6. A method of preparing crystalline Form A of sepiapterin free base having peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 12.4°±0.5, 26.2°±0.5, and 28.9°±0.5 as measured by X-ray diffractometry by irradiation with Cu Kα X-rays, comprising dissolving sepiapterin free base in dimethyl acetamide and exposing the solution to a vapor of dichloromethane and obtaining the crystalline Form A of sepiapterin free base.
7. The method of claim 1 , wherein the Form A of sepiapterin free base has peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 9.5°±0.5, 11.3°±0.5, 12.4°±0.5, 15.6°±0.5, 16.4°±0.5, 26.2°±0.5, 27.2°±0.5, and 28.9°±0.5, as measured by X-ray diffractometry by irradiation with Cu Kα X-rays.
8. The method of claim 1 , wherein Form A of sepiapterin free base has one or more of the following: the X-ray powder diffraction spectrum essentially as shown in FIG. 1 , a loss of weight from 30° C. to 150° C. of less than 15% as measured by thermal gravimetric analysis, an endothermic onset at about 84° C. or about 180° C. in differential scanning calorimetry (DSC) profile, or an endothermic onset at about 84° C. and about 180° C. in differential scanning calorimetry (DSC) profile.
9. The method of claim 2 , wherein the Form A of sepiapterin free base has peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 9.5°±0.5, 11.3°±0.5, 12.4°±0.5, 15.6°±0.5, 16.4°±0.5, 26.2°±0.5, 27.2°±0.5, and 28.9°±0.5, as measured by X-ray diffractometry by irradiation with Cu Kα X-rays.
10. The method of claim 2 , wherein Form A of sepiapterin free base has one or more of the following: the X-ray powder diffraction spectrum essentially as shown in FIG. 1 , a loss of weight from 30° C. to 150° C. of less than 15% as measured by thermal gravimetric analysis, an endothermic onset at about 84° C. or about 180° C. in differential scanning calorimetry (DSC) profile, or an endothermic onset at about 84° C. and about 180° C. in differential scanning calorimetry (DSC) profile.
11. The method of claim 3 , wherein the Form A of sepiapterin free base has peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 9.5°±0.5, 11.3°±0.5, 12.4°±0.5, 15.6°±0.5, 16.4°±0.5, 26.2°±0.5, 27.2°±0.5, and 28.9°±0.5, as measured by X-ray diffractometry by irradiation with Cu Kα X-rays.
12. The method of claim 3 , wherein Form A of sepiapterin free base has one or more of the following: the X-ray powder diffraction spectrum essentially as shown in FIG. 1 , a loss of weight from 30° C. to 150° C. of less than 15% as measured by thermal gravimetric analysis, an endothermic onset at about 84° C. or about 180° C. in differential scanning calorimetry (DSC) profile, or an endothermic onset at about 84° C. and about 180° C. in differential scanning calorimetry (DSC) profile.
13. The method of claim 4 , wherein the Form A of sepiapterin free base has peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 9.5°±0.5, 11.3°±0.5, 12.4°±0.5, 15.6°±0.5, 16.4°±0.5, 26.2°±0.5, 27.2°±0.5, and 28.9°±0.5, as measured by X-ray diffractometry by irradiation with Cu Kα X-rays.
14. The method of claim 4 , wherein the Form A of sepiapterin free base has one or more of the following: the X-ray powder diffraction spectrum essentially as shown in FIG. 1 , a loss of weight from 30° C. to 150° C. of less than 15% as measured by thermal gravimetric analysis, an endothermic onset at about 84° C. or about 180° C. in differential scanning calorimetry (DSC) profile, an endothermic onset at about 84° C. and about 180° C. in differential scanning calorimetry (DSC) profile.
15. The method of claim 5 , wherein the Form A of sepiapterin free base has peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 9.5°±0.5, 11.3°±0.5, 12.4°±0.5, 15.6°±0.5, 16.4°±0.5, 26.2°±0.5, 27.2°±0.5, and 28.9°±0.5, as measured by X-ray diffractometry by irradiation with Cu Kα X-rays.
16. The method of claim 5 , wherein the Form A of sepiapterin free base has one or more of the following: the X-ray powder diffraction spectrum essentially as shown in FIG. 1 , a loss of weight from 30° C. to 150° C. of less than 15% as measured by thermal gravimetric analysis, an endothermic onset at about 84° C. or about 180° C. in differential scanning calorimetry (DSC) profile, or an endothermic onset at about 84° C. and about 180° C. in differential scanning calorimetry (DSC) profile.
17. The method of claim 6 , wherein the Form A of sepiapterin free base has peaks at diffraction angle 2θ (°) of 4.7°±0.5, 7.4°±0.5, 9.5°±0.5, 11.3°±0.5, 12.4°±0.5, 15.6°±0.5, 16.4°±0.5, 26.2°±0.5, 27.2°±0.5, and 28.9°±0.5, as measured by X-ray diffractometry by irradiation with Cu Kα X-rays.
18. The method of claim 6 , wherein the Form A of sepiapterin free base has one or more of the following: the X-ray powder diffraction spectrum essentially as shown in FIG. 1 , a loss of weight from 30° C. to 150° C. of less than 15% as measured by thermal gravimetric analysis, an endothermic onset at about 84° C. or about 180° C. in differential scanning calorimetry (DSC) profile, or an endothermic onset at about 84° C. and about 180° C. in differential scanning calorimetry (DSC) profile.