Pharmaceutical compositions comprising sepiapterin and uses thereof

Abstract

To provide a pharmaceutical composition for treating tetrahydrobiopterin (BH4) deficiency.SOLUTION: A pharmaceutical composition comprises sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, and is stable at room temperature for at least 6 months. The pharmaceutical composition is a solid composition and further comprises an antioxidant, the antioxidant comprising ascorbic acid, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, butylated hydroxytoluene, or butylated hydroxyanisole.SELECTED DRAWING: None

Description

[Background technology]

[0001] Primary tetrahydrobiopterin deficiency (PBD) is caused by deficiencies of GTP cyclohydrolase I (GTP-CH), 6-pyruvoyltetrahydropterin synthase (PTPS), or sepiapterin reductase (SR), which impair tetrahydrobiopterin (BH4) biosynthesis, or by defects in BH4 recycling (pterin-4a-carbinolamine dehydratase (PCD) or dihydropteridine reductase (DHPR) deficiency). PBD accounts for 1-3% of all cases of hyperphenylalaninemia (HPA), with almost all remaining cases being due to phenylalanine hydroxylase deficiency.

[0002] BH4 is an essential cofactor for phenylalanine hydroxylase, tyrosine hydroxylase, tryptophan hydroxylase, fatty acid glyceryl ether oxygenase, and nitric oxide (NO) synthase.

[0003] In PBD, impaired hydroxylation of phenylalanine (Phe) to tyrosine (Tyr) leads to HPA. Decreased Tyr synthesis and decreased activity of Tyr and tryptophan hydroxylase result in decreased formation of neurotransmitters, resulting in neuromotor disorders.

[0004] Phenotypically, BH4 deficiency manifests as deficiency of the HPA and the neurotransmitter precursors L-dopa and 5-hydroxytryptophan, and is detected by screening programs that measure Phe to detect phenylalanine hydroxylase deficiency (the exception being individuals with SR deficiency, who have normal Phe levels).

[0005] Current treatment for BH4 deficiency involves lowering blood Phe levels by oral administration of BH4 (in cases of GTP-CH and PTPS deficiency) and / or a low-Phe diet (primarily in cases of DHPR deficiency), along with administration of the neurotransmitter precursors L-dopa and 5-hydroxytryptophan (5HTP).

[0006] There are few reported long-term follow-up studies of patients with BH4 deficiency. Treatments vary between physicians and clinics, are far from clinically evidence-based, and include pharmacological agents, none of which have been approved for sale for this indication. Thus, there is a need for new treatments for BH4 deficiency. Summary of the Invention

[0007] The present invention features pharmaceutical compositions containing sepiapterin, or pharmaceutically acceptable salts and / or cocrystals thereof, and methods for treating tetrahydrobiopterin-associated disorders using such compositions.

[0008] Accordingly, in one aspect, the invention features a pharmaceutical composition comprising sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, wherein the pharmaceutical composition is stable for at least six months at room temperature (e.g., 25°C, or 25°C and 60% relative humidity). For example, when the composition is stored at room temperature for six months, the level of sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, in the composition decreases by less than 5% (e.g., less than 4%, less than 3%, less than 2%, less than 1%), and / or the level of lactoylpterin in the composition increases by less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%).

[0009] In some embodiments, the pharmaceutical composition is a solid composition (e.g., a powder, capsule, or tablet). In some embodiments, the pharmaceutical composition is formulated for use in a suspension.

[0010] In some embodiments, the pharmaceutical composition further comprises an antioxidant, hi some embodiments, the antioxidant is present in an amount sufficient to stabilize the sepiapterin, or pharmaceutically acceptable salt and / or co-crystal thereof, in the composition at room temperature (e.g., 25°C, or 25°C and 60% relative humidity) for at least 6 months.

[0011] In one aspect, the invention features a solid pharmaceutical composition that includes sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, and an antioxidant. In some embodiments, the antioxidant is present in an amount sufficient to stabilize the sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, in the composition at room temperature (e.g., 25° C., or 25° C. and 60% relative humidity) for at least 6 months.

[0012] In one aspect, the invention features a pharmaceutical composition that includes sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, and an antioxidant (e.g., ascorbic acid), wherein the ratio of sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, to antioxidant is greater than 4:1 wt / wt (e.g., greater than 5:1, greater than 6:1, greater than 7:1, greater than 8:1, greater than 9:1, greater than 10:1, greater than 15:1, or greater than 20:1).

[0013] In one aspect, the invention features a pharmaceutical composition including sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, and an antioxidant. The pharmaceutical composition includes more sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, than the antioxidant, by weight. For example, in some embodiments, the sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, and the antioxidant (e.g., ascorbic acid) are present in a ratio of at least 1:1 (e.g., 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1) wt / wt.

[0014] In some embodiments, the composition is a solid pharmaceutical composition comprising sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, and an antioxidant. In some embodiments, the antioxidant is present in an amount sufficient to stabilize the sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, in the composition for at least 6 months at room temperature (e.g., 25°C, or 25°C and 60% relative humidity). In some embodiments, the ratio of sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, to antioxidant is greater than 4:1 wt / wt (e.g., greater than 5:1, greater than 6:1, greater than 7:1, greater than 8:1, greater than 9:1, greater than 10:1, greater than 15:1, or greater than 20:1).

[0015] In one aspect, the invention features a pharmaceutical composition that includes sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, and less than 10% (e.g., less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%) of an antioxidant (e.g., ascorbic acid) by total weight. In some embodiments, the pharmaceutical composition is substantially free of antioxidants.

[0016] In one aspect, the invention features a pharmaceutical composition comprising sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, and lactoylpterin at less than 50% by weight (e.g., less than 40%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, or less than 0.2%) of the total amount of sepiapterin, or a salt and / or cocrystal thereof, and lactoylpterin in the composition. In some embodiments, the pharmaceutical composition comprises less than 10% lactoylpterin. In some embodiments, the pharmaceutical composition comprises less than 1.3% lactoylpterin. In some embodiments, the pharmaceutical composition further comprises an antioxidant (e.g., ascorbic acid). In other embodiments, the pharmaceutical composition does not include an antioxidant.

[0017] In one aspect, the invention features a particulate pharmaceutical composition comprising sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, and an antioxidant (e.g., ascorbic acid), e.g., for use in a suspension.

[0018] In one aspect, the invention features a pharmaceutical composition formulated as a suspension in a dosing vehicle, comprising a bulking or anti-caking agent, sepiapterin or a pharmaceutically acceptable salt thereof, and an antioxidant, wherein at least 50% (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%) of the sepiapterin, or a pharmaceutically acceptable salt thereof, and / or co-crystal is dissolved in the dosing vehicle.

[0019] In one aspect, the invention features a pharmaceutical composition formulated as a suspension in a dosing vehicle that includes a bulking or anti-caking agent, sepiapterin or a pharmaceutically acceptable salt and / or co-crystal thereof, and an antioxidant, wherein the concentration of sepiapterin or a pharmaceutically acceptable salt and / or co-crystal thereof in the dosing vehicle is between 1 mg / ml and 5 mg / ml (e.g., at least 1 mg / ml, at least 1.1 mg / ml, at least 1.2 mg / ml, at least 1.3 mg / ml, at least 1.4 mg / ml, at least 1.5 mg / ml, at least 1.6 mg / ml, at least 1.7 mg / ml, at least 1.8 mg / ml, at least 1.9 mg / ml, at least 2.0 mg / ml, or at least 2.1 mg / ml).

[0020] In one aspect, the invention features a pharmaceutical composition formulated as a suspension in an administration vehicle that includes a bulking or anti-caking agent, sepiapterin or a pharmaceutically acceptable salt and / or co-crystal thereof, and an antioxidant, wherein at least 50% (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%) of the sepiapterin or a pharmaceutically acceptable salt and / or co-crystal thereof is adsorbed to the bulking or anti-caking agent.

[0021] In some embodiments of any of the above pharmaceutical compositions, the antioxidant is 4-chloro-2,6-di-tert-butylphenol, tocopherol, α-tocopherol, alkylated diphenylamines, ascorbic acid, ascorbyl myristate, ascorbyl palmitate, ascorbyl stearate, β-carotene, butylated hydroxyanisole, butylated hydroxytoluene, citric acid, cysteine, D-α-tocopheryl polyethylene glycol 1000 succinate, deferoxamine methanesulfonate, dodecyl gallate, ethylparaben, folic acid, fumaric acid, gallic acid, glutathione, lecithin, malic acid, The antioxidant is methylparaben, monothioglycerol, N-acetylcysteine, nordihydroguaiaretic acid, octyl gallate, p-phenylenediamine, potassium ascorbate, potassium metabisulfite, potassium sorbate, propionic acid, propyl gallate, retinol, sorbic acid, sodium ascorbate, sodium bisulfite, sodium hydrosulfite, sodium isoascorbate, sodium metabisulfite, sodium sulfite, sodium thiosulfate, tartaric acid, tert-butylhydroquinone, tocopheryl acetate, vitamin A, vitamin B6, vitamin B12, or vitamin E, or a combination thereof. In some embodiments of any of the above pharmaceutical compositions, the antioxidant is ascorbic acid, tocopherol, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, butylated hydroxytoluene, and / or butylated hydroxyanisole.

[0022] In some embodiments of any of the above compositions, the pharmaceutical composition comprises about 20 to 95% (e.g., about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 20 to 30%, about 25 to 45%, about 40 to 60%, about 50 to 75%, about 65 to 75%, about 70 to 90%, or about 85 to 95%) sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, by total weight.

[0023] In some embodiments of any of the above compositions, the pharmaceutical composition further comprises a dispersing agent (e.g., carboxymethylcellulose or a pharmaceutically acceptable salt and / or co-crystal thereof, such as croscarmellose sodium). In some embodiments of any of the above compositions, the pharmaceutical composition comprises 0.1 to 1.5% (e.g., 0.1 to 0.3%, 0.2 to 0.4%, 0.3 to 0.5%, 0.4 to 0.6%, 0.5 to 0.7%, 0.6 to 0.8%, 0.7 to 0.9%, 0.8 to 1%, 0.9 to 1.1%, 0.9 to 1.1%, 1 to 1.2%, 1.1 to 1.3%, 1.2 to 1.4%, or 1.3 to 1.5%) of the dispersing agent (e.g., croscarmellose sodium) by total weight.

[0024] In some embodiments of any of the above compositions, the pharmaceutical composition comprises at least one anti-caking agent or bulking agent (e.g., a bulking agent and an anti-caking agent). In some embodiments, the at least one anti-caking agent or bulking agent is colloidal silicon dioxide or microcrystalline cellulose. In some embodiments, the pharmaceutical composition comprises 60-80%, e.g., 65-75% (e.g., about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 78%, about 79%, or about 80%) of the anti-caking agent and / or bulking agent by total weight. In some embodiments of any of the above compositions, the pharmaceutical composition comprises both colloidal silicon dioxide and microcrystalline cellulose. In some embodiments of any of the foregoing compositions, the pharmaceutical composition comprises 60-65% (e.g., about 60%, about 61%, about 62%, about 63%, about 64%, or about 65%) microcrystalline cellulose by total weight and 2-15% (e.g., about 5-7%, 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%, or about 15%) colloidal silicon dioxide by total weight.

[0025] In some embodiments of any of the above compositions, the sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, is in a crystalline form. In some embodiments, the sepiapterin crystalline form has at least one peak at a diffraction angle 2θ (°) of about 9.7°±0.5, about 10.2°±0.5, and / or about 11.3°±0.5, as measured by X-ray diffraction using CuKα X-ray irradiation or calculated from X-ray diffraction.

[0026] In some embodiments, the sepiapterin crystalline form is characterized by peaks at refraction angles 2θ of at least about 9.7°, about 10.2°, about 11.3°, about 14.0°, about 14.6°, about 19.9°, about 22.2°, about 25.3°, and about 32.4°. In some embodiments, the sepiapterin crystalline form is characterized by peaks at refraction angles 2θ set forth in Table 1.

[0027] [Table 1]

[0028] In other embodiments, the sepiapterin crystalline form has at least one peak at a diffraction angle 2θ (°) of about 8.4°±0.5, about 16.9°±0.5, and / or about 25.4°±0.5, as measured by X-ray diffraction using CuKα X-ray irradiation or calculated from X-ray diffraction.

[0029] In some embodiments, the sepiapterin crystalline form is characterized by refraction at refraction angles 2θ of at least about 8.4°, about 14.9°, about 16.9°, about 25.4°, and about 34.1°. In some embodiments, the sepiapterin crystalline form is characterized by refraction at refraction angles 2θ set forth in Table 2.

[0030] [Table 2]

[0031] In further embodiments, the sepiapterin crystalline form has at least one peak at a diffraction angle 2θ (°) of about 5.7°±0.5, about 7.8°±0.5, and / or about 25.4°±0.5, as measured by X-ray diffraction using CuKα X-ray irradiation or calculated from X-ray diffraction.

[0032] In some embodiments, the sepiapterin crystalline form is characterized by refraction at refraction angles 2θ of at least about 5.7°, about 7.8°, about 9.1°, about 11.5°, about 15.3°, about 16.0°, about 20.1°, about 25.4°, and about 26.6°. In some embodiments, the sepiapterin crystalline form is characterized by refraction at refraction angles 2θ set forth in Table 3.

[0033] [Table 3]

[0034] In yet other embodiments, the sepiapterin crystalline form has at least one peak at a diffraction angle 2θ (°) of about 8.9°±0.5, about 10.3°±0.5, and / or about 26.0°±0.5, as measured by X-ray diffraction using CuKα X-ray irradiation or calculated from X-ray diffraction.

[0035] In some embodiments, the sepiapterin crystalline form is characterized by refraction at refraction angles 2θ of at least about 8.9°, about 10.3°, about 10.9°, about 17.8°, about 24.9°, about 26.0°, about 26.7°, about 26.8°, and about 28.3°. In some embodiments, the sepiapterin crystalline form is characterized by refraction at refraction angles 2θ set forth in Table 4.

[0036] [Table 4]

[0037] In yet other embodiments, the sepiapterin crystalline form has at least one peak at diffraction angles 2θ of 10.0°±0.5, 10.6°±0.5, and 25.7°±0.5.

[0038] In some embodiments, the sepiapterin crystalline form is characterized by refractive indices at refractive index 2θ angles of at least 10.0°±0.5, 10.6°±0.5, 11.2°±0.5, 15.3°±0.5, 15.9°±0.5, 22.8°±0.5, 24.4°±0.5, 25.0°±0.5, 25.7°±0.5, and 26.6°±0.5. In some embodiments, the sepiapterin crystalline form is characterized by refractive indices at refractive index angles 2θ set forth in Table 5.

[0039] [Table 5]

[0040] In some embodiments of any of the foregoing compositions, the sepiapterin is a pharmaceutically acceptable salt and / or co-crystal of sepiapterin. In some embodiments, the salt and / or co-crystal of sepiapterin is a methanesulfonate salt and / or co-crystal, a nicotine salt and / or co-crystal, a p-toluenesulfonate salt and / or co-crystal, a benzenesulfonate salt and / or co-crystal, a phosphate salt and / or co-crystal (e.g., a 1:1 phosphate salt and / or co-crystal, i.e., one molecule of sepiapterin for one molecule of phosphoric acid), a malonate salt and / or co-crystal (e.g., a 1:1 malonate salt and / or co-crystal, i.e., one molecule of sepiapterin for one molecule of malonic acid), a tartrate salt and / or co-crystal (e.g., a 1:1 tartrate salt and / or co-crystal, i.e., one molecule of sepiapterin for one molecule of tartaric acid). pterin), gentisate salts and / or co-crystals (e.g., 2:1 gentisate salts and / or co-crystals, i.e., 1 molecule of gentisate to 2 molecules of sepiapterin), fumarate salts and / or co-crystals (e.g., 2:1 fumarate salts and / or co-crystals, i.e., 2 molecules of sepiapterin to 1 molecule of fumaric acid), glycolate salts and / or co-crystals (e.g., 3:1 glycolate salts and / or co-crystals, i.e., 3 molecules of sepiapterin to 1 molecule of glycolic acid), acetate salts and / or co-crystals, or sulfate salts and / or co-crystals (e.g., 2:1 sulfate salts and / or co-crystals, i.e., 1 molecule of sulfate to 2 molecules of sepiapterin). In some embodiments, the salt and / or co-crystal is a salt with an acid. In some embodiments, the salt and / or co-crystal is a co-crystal with an acid. In some embodiments, the salts and / or co-crystals of sepiapterin have improved properties, for example, improved stability, purity, exposure, and / or bioavailability.

[0041] In some embodiments, the salt and / or co-crystal of sepiapterin is a methanesulfonate salt and / or co-crystal.

[0042] In some embodiments, the salt and / or co-crystal of sepiapterin is a nicotinic acid salt and / or co-crystal.

[0043] In some embodiments, the salt and / or co-crystal of sepiapterin is a p-toluenesulfonate salt and / or co-crystal.

[0044] In some embodiments, the salt and / or co-crystal of sepiapterin is a benzenesulfonate salt and / or co-crystal.

[0045] In some embodiments, the salts and / or co-crystals of sepiapterin are phosphate salts and / or co-crystals, and the phosphate counterion is H2PO4 - In some embodiments, the salt and / or co-crystal of sepiapterin is a phosphate salt and / or co-crystal, wherein the phosphate counterion is HPO 2- In some embodiments, the salt and / or co-crystal of sepiapterin is a phosphate salt and / or co-crystal, and the phosphate counterion is PO4 3- In some embodiments, the salt and / or co-crystal of sepiapterin is a 1:1 phosphate and / or co-crystal, i.e., one molecule of sepiapterin for one molecule of phosphate. In some embodiments, the salt and / or co-crystal of sepiapterin is a 2:1 phosphate and / or co-crystal, i.e., two molecules of sepiapterin for one molecule of phosphate.

[0046] In some embodiments, the salts and / or co-crystals of sepiapterin are 1:1 malonate salts and / or co-crystals, i.e., one molecule of sepiapterin for every molecule of malonic acid, hi some embodiments, the salts and / or co-crystals of sepiapterin are 2:1 malonate salts and / or co-crystals, i.e., two molecules of sepiapterin for every molecule of malonic acid.

[0047] In some embodiments, the salt and / or co-crystal of sepiapterin is a 1:1 tartrate salt and / or co-crystal, i.e., one molecule of sepiapterin for every molecule of tartaric acid, hi some embodiments, the salt and / or co-crystal of sepiapterin is a 2:1 tartrate salt and / or co-crystal, i.e., two molecules of sepiapterin for every molecule of tartaric acid.

[0048] In some embodiments, the sepiapterin salt and / or co-crystal is a 1:1 gentisate salt and / or co-crystal, i.e., one molecule of gentisate to one molecule of sepiapterin, hi some embodiments, the sepiapterin salt and / or co-crystal is a 2:1 gentisate salt and / or co-crystal, i.e., two molecules of sepiapterin to one molecule of gentisate.

[0049] In some embodiments, the salt and / or co-crystal of sepiapterin is a 1:1 fumarate salt and / or co-crystal, i.e., one molecule of sepiapterin for every molecule of fumaric acid, hi some embodiments, the salt and / or co-crystal of sepiapterin is a 2:1 fumarate salt and / or co-crystal, i.e., two molecules of sepiapterin for every molecule of fumaric acid.

[0050] In some embodiments, the salt and / or co-crystal of sepiapterin is a 1:1 glycolate and / or co-crystal, i.e., one molecule of sepiapterin for one molecule of glycolic acid. In some embodiments, the salt and / or co-crystal of sepiapterin is a 2:1 glycolate and / or co-crystal, i.e., two molecules of sepiapterin for one molecule of glycolic acid. In some embodiments, the salt and / or co-crystal of sepiapterin is a 3:1 glycolate and / or co-crystal, i.e., three molecules of sepiapterin for one molecule of glycolic acid.

[0051] In some embodiments, the salt and / or co-crystal of sepiapterin is a sulfate salt and / or co-crystal, and the sulfate counterion is HSO - In some embodiments, the salt and / or co-crystal of sepiapterin is a sulfate salt and / or co-crystal, and the sulfate counterion is SO 2- In some embodiments, the salt and / or co-crystal of sepiapterin is a 1:1 sulfate salt and / or co-crystal, i.e., one molecule of sepiapterin for every molecule of sulfate. In some embodiments, the salt and / or co-crystal of sepiapterin is a 2:1 sulfate salt and / or co-crystal, i.e., two molecules of sepiapterin for every molecule of sulfate.

[0052] In some embodiments, the salt and / or co-crystal of sepiapterin is an acetate salt and / or co-crystal.

[0053] In some embodiments, the pharmaceutically acceptable salt and / or co-crystal is crystalline. In some embodiments, the pharmaceutically acceptable salt and / or co-crystal contains less than 40% by weight of amorphous compound (e.g., less than 30%, less than 20%, less than 10%, less than 5%, less than 1%, or between 30-40%, 25-35%, 20-30%, 15-25%, 10-20%, 5-15%, or 1-10%). In some embodiments, the pharmaceutically acceptable salt and / or co-crystal is substantially free of amorphous compound. In some embodiments, the pharmaceutically acceptable salt and / or co-crystal is substantially free of other salts and / or co-crystals or crystalline forms of sepiapterin.

[0054] In some embodiments, the pharmaceutically acceptable salt and / or co-crystal of sepiapterin is a hydrochloride salt and / or co-crystal. In some embodiments, the hydrochloride salt and / or co-crystal has an endothermic onset temperature of about 218°C (e.g., 216°C to 220°C, e.g., 217°C to 219°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the hydrochloride salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0055] In some embodiments, the hydrochloride salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 7.8±0.5, as measured by powder X-ray diffraction. In some embodiments, the hydrochloride salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 7.8±0.5, 12.9±0.5, and / or 26.2±0.5, as measured by powder X-ray diffraction. In some embodiments, the hydrochloride salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 6, as measured by powder X-ray diffraction. In some embodiments, the hydrochloride salt and / or co-crystal has all of the peaks listed in Table 6, as measured by powder X-ray diffraction.

[0056] [Table 6]

[0057] In some embodiments, the pharmaceutically acceptable salt and / or co-crystal of sepiapterin is a methanesulfonate salt and / or co-crystal. In some embodiments, the methanesulfonate salt and / or co-crystal has an endothermic onset temperature of about 182°C (e.g., 180°C to 184°C, e.g., 181°C to 183°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the methanesulfonate salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0058] In some embodiments, the methanesulfonate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 23.5±0.5, as measured by powder X-ray diffraction. In some embodiments, the methanesulfonate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 7.9±0.5, 23.5±0.5, and / or 29.0±0.5, as measured by powder X-ray diffraction. In some embodiments, the methanesulfonate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 7, as measured by powder X-ray diffraction. In some embodiments, the methanesulfonate salt and / or co-crystal has all of the peaks listed in Table 7, as measured by powder X-ray diffraction.

[0059] [Table 7]

[0060] In some embodiments, the methanesulfonate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 7.9±0.5, as measured by powder X-ray diffraction. In some embodiments, the methanesulfonate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 7.9±0.5, 23.4±0.5, and / or 28.9±0.5, as measured by powder X-ray diffraction. In some embodiments, the methanesulfonate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 8, as measured by powder X-ray diffraction. In some embodiments, the methanesulfonate salt and / or co-crystal has all of the peaks listed in Table 8, as measured by powder X-ray diffraction.

[0061] [Table 8]

[0062] In some embodiments, the pharmaceutically acceptable salt and / or co-crystal of sepiapterin is a nicotinate salt and / or co-crystal. In some embodiments, the nicotinate salt and / or co-crystal has an endothermic onset temperature of about 220°C (e.g., 218°C to 222°C, e.g., 219°C to 221°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the nicotinate salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0063] In some embodiments, the nicotinate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 24.5±0.5, as measured by powder X-ray diffraction. In some embodiments, the nicotinate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 9.9±0.5, 23.2±0.5, and / or 24.5±0.5, as measured by powder X-ray diffraction. In some embodiments, the nicotinate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 9, as measured by powder X-ray diffraction. In some embodiments, the nicotinate salt and / or co-crystal has all of the peaks listed in Table 9, as measured by powder X-ray diffraction.

[0064] [Table 9]

[0065] In some embodiments, the sepiapterin salt and / or co-crystal is a toluenesulfonate salt and / or co-crystal. In some embodiments, the toluenesulfonate salt and / or co-crystal has an endothermic onset temperature of about 190°C (e.g., 188°C to 192°C, e.g., 189°C to 191°C) and / or 263°C (e.g., 261°C to 265°C, 262°C to 264°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the toluenesulfonate salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0066] In some embodiments, the toluenesulfonate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 6.5±0.5, as measured by powder X-ray diffraction. In some embodiments, the toluenesulfonate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 6.5±0.5, 15.1±0.5, and / or 23.4±0.5, as measured by powder X-ray diffraction. In some embodiments, the toluenesulfonate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 10, as measured by powder X-ray diffraction. In some embodiments, the toluenesulfonate salt and / or co-crystal has all of the peaks listed in Table 10, as measured by powder X-ray diffraction.

[0067] [Table 10]

[0068] In some embodiments, the salt and / or co-crystal of sepiapterin is a benzenesulfonate salt and / or co-crystal. In some embodiments, the benzenesulfonate salt and / or co-crystal has an endothermic onset temperature of about 193°C (e.g., 191°C to 195°C, e.g., 192°C to 194°C) and / or 206°C (e.g., 204°C to 208°C, 205°C to 207°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the benzenesulfonate salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0069] In some embodiments, the benzenesulfonate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 6.5±0.5 as measured by powder X-ray diffraction. In some embodiments, the benzenesulfonate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 6.5±0.5, 14.8±0.5, and / or 19.6±0.5 as measured by powder X-ray diffraction. In some embodiments, the benzenesulfonate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 11 as measured by powder X-ray diffraction. In some embodiments, the benzenesulfonate salt and / or co-crystal has all of the peaks listed in Table 11 as measured by powder X-ray diffraction.

[0070] [Table 11]

[0071] In some embodiments, the salt and / or co-crystal of sepiapterin is a sulfate salt. In some embodiments, the sulfate salt and / or co-crystal has an endothermic onset temperature of about 196°C (e.g., 194°C to 198°C, e.g., 195°C to 197°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the sulfate salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0072] In some embodiments, the sulfate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 5.1±0.5, as measured by powder X-ray diffraction. In some embodiments, the sulfate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 5.1±0.5, 7.8±0.5, and / or 23.0±0.5, as measured by powder X-ray diffraction. In some embodiments, the sulfate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 12, as measured by powder X-ray diffraction. In some embodiments, the sulfate salt and / or co-crystal has all of the peaks listed in Table 12, as measured by powder X-ray diffraction.

[0073] [Table 12]

[0074] In some embodiments, the pharmaceutically acceptable salt and / or co-crystal of sepiapterin is a phosphate and / or co-crystal. In some embodiments, the phosphate and / or co-crystal has an endothermic onset temperature of about 144°C (e.g., 142°C to 146°C, e.g., 143°C to 145°C) and / or 207°C (e.g., 205°C to 209°C, 206°C to 208°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the phosphate and / or co-crystal exhibits a weight loss of less than 12% (e.g., less than 10%, less than 5%, less than 2%, or less than 1%) from 31°C to 150°C, as measured by thermogravimetric analysis.

[0075] In some embodiments, the phosphate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 25.6±0.5, as measured by powder X-ray diffraction. In some embodiments, the phosphate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 16.6±0.5, 22.2±0.5, and / or 25.6±0.5, as measured by powder X-ray diffraction. In some embodiments, the phosphate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 13, as measured by powder X-ray diffraction. In some embodiments, the phosphate salt and / or co-crystal has all of the peaks listed in Table 13, as measured by powder X-ray diffraction.

[0076] [Table 13]

[0077] In some embodiments, the salt and / or co-crystal of sepiapterin is a malonate and / or co-crystal. In some embodiments, the malonate and / or co-crystal has an endothermic onset temperature of about 175°C (e.g., 173°C to 177°C, e.g., 174°C to 176°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the malonate and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0078] In some embodiments, the malonate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 6.9±0.5 as measured by powder X-ray diffraction. In some embodiments, the malonate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 6.9±0.5, 23.8±0.5, and / or 25.5±0.5 as measured by powder X-ray diffraction. In some embodiments, the malonate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 14 as measured by powder X-ray diffraction. In some embodiments, the malonate salt and / or co-crystal has all of the peaks listed in Table 14 as measured by powder X-ray diffraction.

[0079] [Table 14]

[0080] In some embodiments, the salt and / or co-crystal of sepiapterin is a tartrate salt and / or co-crystal (e.g., an L-tartrate salt and / or co-crystal). In some embodiments, the tartrate salt and / or co-crystal has an endothermic onset temperature of about 156°C (e.g., 154°C to 158°C, e.g., 155°C to 157°C) and / or 175°C (e.g., 173°C to 177°C, e.g., 174°C to 176°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the tartrate salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C as measured by thermogravimetric analysis.

[0081] In some embodiments, the tartrate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 7.4±0.5, as measured by powder X-ray diffraction. In some embodiments, the tartrate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 7.4±0.5, 21.8±0.5, and / or 23.9±0.5, as measured by powder X-ray diffraction. In some embodiments, the tartrate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 15, as measured by powder X-ray diffraction. In some embodiments, the tartrate salt and / or co-crystal has all of the peaks listed in Table 15, as measured by powder X-ray diffraction.

[0082] [Table 15]

[0083] In some embodiments, the sepiapterin salt and / or co-crystal is a fumarate salt and / or co-crystal. In some embodiments, the fumarate salt and / or co-crystal has an endothermic onset temperature of about 77°C (e.g., 75°C to 79°C, e.g., 76°C to 78°C), 133°C (e.g., 131°C to 135°C, e.g., 132°C to 134°C), and / or 190°C (e.g., 188°C to 192°C, e.g., 189°C to 191°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the fumarate salt and / or co-crystal exhibits a weight loss of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C, as measured by thermogravimetric analysis.

[0084] In some embodiments, the fumarate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 24.0±0.5, as measured by powder X-ray diffraction. In some embodiments, the fumarate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 11.4±0.5, 11.9±0.5, and / or 24.0±0.5, as measured by powder X-ray diffraction. In some embodiments, the fumarate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 16, as measured by powder X-ray diffraction. In some embodiments, the fumarate salt and / or co-crystal has all of the peaks listed in Table 16, as measured by powder X-ray diffraction.

[0085] [Table 16]

[0086] In some embodiments, the salt of sepiapterin is a gentisate salt and / or co-crystal. In some embodiments, the gentisate salt and / or co-crystal has an endothermic onset temperature of about 83°C (e.g., 81°C to 85°C, e.g., 82°C to 84°C), 134°C (e.g., 132°C to 136°C, e.g., 133°C to 135°C), and / or 149°C (e.g., 147°C to 151°C, e.g., 148°C to 150°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the gentisate salt and / or co-crystal exhibits a weight loss of less than 7% (e.g., less than 5%, less than 3%, less than 2%, or less than 1%) from 31°C to 150°C, as measured by thermogravimetric analysis.

[0087] In some embodiments, the gentisate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 7.1±0.5 as measured by powder X-ray diffraction. In some embodiments, the gentisate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 7.1±0.5, 8.7±0.5, and / or 26.7±0.5 as measured by powder X-ray diffraction. In some embodiments, the gentisate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 17 as measured by powder X-ray diffraction. In some embodiments, the gentisate salt and / or co-crystal has all of the peaks listed in Table 17 as measured by powder X-ray diffraction.

[0088] [Table 17]

[0089] In some embodiments, the salt of sepiapterin is a glycolate salt and / or a co-crystal. In some embodiments, the glycolate salt and / or co-crystal has an endothermic onset temperature of about 79°C (e.g., 77°C to 81°C, e.g., 78°C to 80°C), 90°C (e.g., 88°C to 92°C, e.g., 89°C to 91°C), 132°C (e.g., 130°C to 134°C, e.g., 131°C to 133°C), and / or 152°C (e.g., 130°C to 134°C, e.g., 131°C to 133°C) in a differential scanning calorimetry (DSC) profile. In some embodiments, the glycolate salt and / or co-crystal exhibits a weight loss of less than 21% (e.g., less than 15%, less than 10%, less than 5%, or less than 1%) from 31°C to 150°C, as measured by thermogravimetric analysis.

[0090] In some embodiments, the glycolate salt and / or co-crystal has at least one peak at a diffraction angle 2θ (°) of 7.6±0.5, as measured by powder X-ray diffraction. In some embodiments, the glycolate salt and / or co-crystal further has at least one peak at a diffraction angle 2θ (°) of 7.6±0.5, 10.7±0.5, and / or 24.0±0.5, as measured by powder X-ray diffraction. In some embodiments, the glycolate salt and / or co-crystal has one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more) peaks listed in Table 18, as measured by powder X-ray diffraction. In some embodiments, the glycolate salt and / or co-crystal has all of the peaks listed in Table 18, as measured by powder X-ray diffraction.

[0091] [Table 18]

[0092] In some embodiments of any of the foregoing compositions, the sepiapterin or its salts and / or cocrystals are formulated in particles less than 200 μm in size (e.g., less than 180 μm, less than 160 μm, less than 140 μm, less than 120 μm, less than 100 μm, or less than 80 μm).

[0093] In some embodiments of any of the foregoing compositions, the pharmaceutical composition is formulated as particles (e.g., particles for use in a suspension). In some embodiments, the particles are less than 200 μm in size (e.g., less than 180 μm, less than 160 μm, less than 140 μm, less than 120 μm, less than 100 μm, or less than 80 μm).

[0094] In some embodiments of any of the foregoing compositions, the pharmaceutical composition comprises less than 50% by weight of lactoylpterin (e.g., less than 40%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.3%, or less than 0.2%) of the combined amount of sepiapterin or its salt(s) and / or cocrystal(s) and lactoylpterin in the composition. In some embodiments, the pharmaceutical composition comprises less than 1.3% lactoylpterin.

[0095] In some embodiments of any of the foregoing compositions, the pharmaceutical composition further comprises a dosage vehicle (eg, a dosage vehicle having a viscosity of about 50 to 1750 centipoise).

[0096] In some embodiments of any of the foregoing compositions, the pharmaceutical composition, upon administration to a subject, results in greater (e.g., at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 2-fold, at least 3-fold, or at least 4-fold) plasma or hepatocyte levels of tetrahydrobiopterin compared to that resulting from administration of a pharmaceutical composition containing the same amount of tetrahydrobiopterin (e.g., as measured by Tmax, Cmax, AUC, or plasma concentration 15 minutes after administration).

[0097] In some embodiments of any of the foregoing compositions, the pharmaceutical composition, upon administration to a subject, results in a plasma or hepatocyte level of tetrahydrobiopterin that is greater (e.g., at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 2-fold, at least 3-fold, or at least 4-fold) than that resulting from administration of a pharmaceutical composition comprising the same amount of sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof and 10% or more antioxidant (e.g., as measured by Tmax, Cmax, AUC, or plasma concentration 15 minutes after administration).

[0098] In one aspect, the invention features a method for producing a pharmaceutical composition comprising sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof. The method includes: a) mixing microcrystalline cellulose and / or colloidal silicon dioxide; b) adding sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, a dispersing agent, and / or an antioxidant to the mixture of step a; and c) mixing the microcrystalline cellulose, colloidal silicon dioxide, sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, a dispersing agent, and / or an antioxidant, thereby producing a pharmaceutical composition comprising sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof. In some embodiments, steps a), b), and c) may be performed in any order.

[0099] In some embodiments, the mixture of microcrystalline cellulose and colloidal silicon dioxide is passed through a filter having pores of less than 200 μm (e.g., less than 180 μm, less than 160 μm, less than 140 μm, less than 120 μm, less than 100 μm, or less than 80 μm) prior to step b.

[0100] In some embodiments, the mixture of microcrystalline cellulose, colloidal silicon dioxide, sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, dispersing agent, and antioxidant is passed through a filter having pores of less than 200 μm (e.g., less than 180 μm, less than 160 μm, less than 140 μm, less than 120 μm, less than 100 μm, or less than 80 μm).

[0101] In some embodiments of any of the foregoing compositions, the weight percentages are measured for the dry composition (e.g., before suspension in a liquid such as water). In some embodiments, if sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, is present in the composition as a pharmaceutically acceptable salt and / or cocrystal, the weight is the weight of the sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, not including the counterion.

[0102] In some embodiments of any of the above methods, the antioxidant is ascorbic acid. In some embodiments of any of the above methods, the dispersing agent is croscarmellose sodium.

[0103] In one aspect, the invention features a method for treating a tetrahydrobiopterin-associated disorder (e.g., phenylketonuria or tetrahydrobiopterin deficiency) in a subject in need thereof, the method including administering an effective amount of any of the pharmaceutical compositions described above.

[0104] In one aspect, the invention features a method of increasing tetrahydrobiopterin levels in a subject in need thereof, the method including administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0105] In one aspect, the invention features a method of reducing phenylalanine levels in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0106] In one aspect, the invention features a method for increasing the activity of phenylalanine hydroxylase in a subject, the method including administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0107] In one aspect, the invention features a method of treating phenylketonuria in a subject in need thereof, the method including administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0108] In one aspect, the invention features a method of treating gastroparesis in a subject in need thereof, the method including administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0109] In one aspect, the invention features a method of increasing serotonin levels in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0110] In one aspect, the invention features a method of increasing tryptophan hydroxylase activity in a subject, the method including administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0111] In one aspect, the invention features a method of increasing dopamine levels in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0112] In one aspect, the invention features a method for increasing tyrosine hydroxylase activity in a subject, the method including administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0113] In one aspect, the invention features a method of increasing nitric oxide synthase activity in a subject, the method including administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0114] In one aspect, the invention features a method for increasing alkylglycerol monooxygenase activity in a subject, comprising administering to the subject an effective amount of any of the pharmaceutical compositions described above.

[0115] In some embodiments of any of the methods, the effective amount of any of the pharmaceutical compositions comprises an amount sufficient to increase the level of tetrahydrobiopterin in the subject's plasma one hour after administration by at least 2-fold (e.g., at least 3, 4, 5, 6, 7, 9, or 10-fold) compared to the level of tetrahydrobiopterin before administration.

[0116] In some embodiments of any of the methods, the effective amount of any of the pharmaceutical compositions comprises an amount sufficient to increase the level of tetrahydrobiopterin in the CSF and / or brain of the subject by at least 2-fold (e.g., at least 3, 4, 5, 6, 7, 9, or 10-fold) one hour after administration compared to the level of tetrahydrobiopterin before administration.

[0117] In some embodiments of any of the above methods, the subject is a human. In some embodiments of any of the above methods, the method comprises combining any of the above pharmaceutical compositions with an administration vehicle prior to administration.

[0118] definition In this application, unless otherwise clear from the context, (i) the term "a" may be understood to mean "at least one," (ii) the term "or" may be understood to mean "and / or," (iii) the terms "comprise" and "including" may be understood to encompass the listed elements or steps, whether presented by themselves or with one or more additional elements or steps, and (iv) the terms "about" and "approximately" may be understood to allow for standard variations as understood by one of ordinary skill in the art, and (v) when ranges are provided, the endpoints are included.

[0119] As used herein, the term "administration" refers to the administration of a composition to a subject. Administration to an animal subject (e.g., administration to a human) can be by any suitable route. For example, in one embodiment, administration includes bronchial (including bronchial instillation), buccal, enteral, interdermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, intratracheal (including intratracheal instillation), transdermal, intravaginal, or intravitreal.

[0120] The term "anti-caking agent" refers to an additive added to powdered or granular pharmaceutical formulations to prevent the formation of lumps. Exemplary anti-caking agents include colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, talcum powder, sodium aluminosilicate, potassium aluminum silicate, calcium aluminosilicate, bentonite, aluminum silicate, stearic acid, and polydimethylsiloxane.

[0121] The term "antioxidant" refers to an active pharmaceutical ingredient that can minimize the oxidative degradation of an active pharmaceutical ingredient. Examples of antioxidants include ascorbic acid, tocopherol, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, ethylenediaminetetraacetic acid, sodium bisulfite, sodium metabisulfite, thiourea, butylated hydroxytoluene, butylated hydroxyanisole, vitamin E, 4-chloro-2,6-di-tert-butylphenol, alkylated diphenylamines, ascorbyl myristate, ascorbyl stearate, beta-carotene, citric acid, cysteine, D-alpha-tocopheryl polyethylene glycol 1000 succinate, and deferoxamine methanesulfonate. Ingredients include: methylparaben, dodecyl gallate, ethylparaben, folic acid, fumaric acid, gallic acid, lecithin, malic acid, methylparaben, monothioglycerol, nordihydroguaiaretic acid, octyl gallate, p-phenylenediamine, potassium ascorbate, potassium metabisulfite, potassium sorbate, propionic acid, propyl gallate, sorbic acid, sodium ascorbate, sodium hydrosulfite, sodium isoascorbate, sodium sulfite, sodium thiosulfate, tartaric acid, tert-butylhydroquinone, tocopheryl acetate, vitamin A, vitamin B6, and vitamin B12.

[0122] As used herein, the term "BH4-related disease" or "tetrahydrobiopterin-associated disorder" refers to any disease or disorder that may derive therapeutic benefit from modulating (e.g., increasing) levels of BH4, e.g., phenylketonuria.

[0123] "Measuring the level of a compound" means detecting the compound directly or indirectly by methods known in the art. "Directly measuring" means performing a process to obtain a physical entity or value (e.g., performing an assay or test on a sample, or "analyzing a sample" as that term is defined herein). "Indirectly measuring" means receiving a physical entity or value from another party or source (e.g., a third-party testing laboratory that directly obtains the physical entity or value). Methods for measuring compound levels generally include, but are not limited to, liquid chromatography (LC)-mass spectrometry.

[0124] The term "dispersing agent" refers to an agent used in pharmaceutical formulations to separate particles in the formulation, e.g., release its active ingredient upon contact with moisture. Examples include cross-linked polyvinylpyrrolidone, carboxymethylcellulose (e.g., croscarmellose salts, e.g., croscarmellose sodium), starch (e.g., sodium starch glycolate), or alginic acid.

[0125] An "effective amount" of a compound may vary depending on factors such as the individual's medical condition, age, sex, weight, and the ability of the compound to elicit a desired response. A therapeutically effective amount includes an amount in which any toxic or detrimental effects of the compound are outweighed by the therapeutically beneficial effects. A therapeutically effective amount also includes an amount sufficient to provide a benefit, e.g., a clinical benefit.

[0126] "Increasing the activity" of an enzyme means increasing the level of an activity associated with the enzyme, e.g., phenylalanine hydroxylase, or an associated downstream effect. A non-limiting example of increasing the activity of an enzyme includes increasing the activity of phenylalanine hydroxylase, which results in a decrease in the level of phenylalanine. The activity level of an enzyme can be measured using any method known in the art.

[0127] "Level" refers to the level of a compound as compared to a reference. The reference can be any useful reference, as defined herein. A "decreased level" or "increased level" of a compound refers to a decrease or increase in the level of a compound as compared to a reference (e.g., 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%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more decrease or increase, as compared to a reference. and a decrease or increase of greater than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, about a 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, or less, or about a 1.2-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold, or more. The level of the compound may be expressed as mass / volume (e.g., g / dL, mg / ml, μg / ml, ng / ml) or as a relative percentage of the total compounds in the sample.

[0128] As used herein, the term "pharmaceutical composition" refers to a composition comprising a compound described herein formulated with a pharmaceutically acceptable excipient. A pharmaceutical composition may be manufactured or sold with the approval of a government regulatory agency as part of a therapeutic regimen for the treatment of a disease in a mammal. A pharmaceutical composition can be formulated, for example, for oral administration in a unit dosage form (e.g., a tablet, capsule, caplet, gelcap, suspension, solution, or syrup), for topical administration (e.g., a cream, gel, lotion, or ointment), for intravenous administration (e.g., a sterile solution in a solvent system suitable for intravenous use, free of particulate matter), or any other pharmaceutically acceptable formulation.

[0129] The term "pharmaceutically acceptable salt" as used herein refers to any pharmaceutically acceptable salt of sepiapterin. Pharmaceutically acceptable salts include ion pairs of sepiapterin in the solid state and / or in solution. Pharmaceutically acceptable cocrystals include free base sepiapterin and an acid in the solid state. A mixture of salt forms and cocrystal forms may be present in the same composition. For example, pharmaceutically acceptable salts of sepiapterin include those that are within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without undue toxicity, irritation, or allergic reaction, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Remington: The Science and Practice of Pharmacy, (22nd ed.) ed. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA). The salts can be prepared in situ during the final isolation and purification of the compounds described herein, or can be prepared separately by reacting the free base group with a suitable organic acid.

[0130] Sepiapterin can be prepared as pharmaceutically acceptable salts and / or co-crystals. These salts may be acid addition salts, including inorganic or organic acids. Suitable pharmaceutically acceptable acids and methods for preparing suitable salts are well known in the art.

[0131] Representative acid addition salts include 4-acetamidobenzoate, acetate, adipate, alginate, 4-aminosalicylate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, carbonate, cinnimate, citrate, cyclopentanepropionate, cyclamate, decanoate, 2,2,2-dichloroacetate, digluconate, dodecyl sulfate, ethane-1,2-disulfonate, ethanesulfonate, formate, fumarate, galactarate, gentisate, glucoheptonate, gluconate, glucoronate, glutamate, glutarate, glycerophosphate, glycolate, hemisulfate, heptonate, hexanoate, hippurate, hydrobromide, and the like. The following salts include acetone, hydrochloride, hydrochloride, 1-hydroxy-2-naphthoate, 2-hydroxyethanesulfonate, isobutyrate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, mandolate, methanesulfonate, naphthalene-1,5-disulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, octanoate, oleate, oxaate, 2-oxoglutarate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyroglutamate, salicylate, sebacate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate salts.

[0132] "Reference" refers to a useful reference used to compare the level of a compound. A reference can be any sample, standard, standard curve, or level used for comparison purposes. A reference can be a normal reference sample or reference standard or level. A "reference sample" can be, for example, a control, e.g., a predetermined negative control value such as a "normal control," or a previous sample taken from the same subject; a sample taken from a normal healthy subject, such as normal cells or normal tissue; a sample (e.g., cell or tissue) from a subject without a disease; a sample from a subject diagnosed with a disease but not yet treated with a compound of the invention; a sample from a subject treated with a compound of the invention; or a sample of a purified compound (e.g., any of those described herein) at a known normal concentration. A "reference standard or level" refers to a value or numerical value obtained from a reference sample. A "normal control value" is a predetermined value indicative of a non-disease state, e.g., a value expected in a healthy control subject. Typically, a normal control value is expressed as a range ("between X and Y"), a high threshold ("not higher than X"), or a low threshold ("not lower than X"). Subjects with measurements within the normal control value for a particular biomarker are typically referred to as "within the normal range" for that biomarker. Normal reference values ​​or levels can be values ​​or values ​​derived from normal subjects who do not have a disease or disorder (e.g., cancer) and who have been treated with a compound of the invention. In preferred embodiments, the reference sample, standard, or level is matched to the subject sample in at least one of the following criteria: age, weight, sex, disease stage, and overall health. A standard curve of levels of a purified compound (e.g., any of those described herein) within the normal reference range can also be used as a reference.

[0133] As used herein, the term "subject" or "patient" refers to any organism to which a composition of the invention can be administered, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may be a human or animal seeking or in need of treatment, undergoing treatment, or potentially undergoing treatment, or receiving care by a trained professional for a particular disease or condition.

[0134] As used herein, the terms "treat," "treated," or "treating" refer to both therapeutic and prophylactic or preventative treatments, where the objective is to prevent or slow (alleviate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical result. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in the extent of a condition, disorder, or disease, stabilization (i.e., not worsening) of a condition, disorder, or disease, delay in the onset or slowing of the progression of a condition, disorder, or disease, improvement or reversal (whether partial or total) of the state of a condition, disorder, or disease (whether detectable or undetectable), improvement in at least one measurable physical parameter (not necessarily discernible by the patient), or enhancement or amelioration of a condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival compared to expected survival if not receiving treatment.

[0135] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present disclosure; however, other suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0136] The details of one or more embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will become apparent from the specification and claims. [Brief explanation of the drawings]

[0137] [Figure 1] FIG. 1 is a table showing the protocol for pharmacokinetic testing of the pharmaceutical composition of the present invention. [Figure 2] FIG. 2 is a graph showing the plasma concentration of sepiapterin after administration of the pharmaceutical composition of the present invention. [Figure 3] FIG. 3 is a table showing the concentrations of tetrahydrobiopterin in the kidney, liver, and urine upon treatment with tetrahydrobiopterin or sepiapterin. [Figure 4] FIG. 4 is an IR spectrum of the free base of sepiapterin. [Figure 5] FIG. 5 shows IR spectra of sepiapterin hydrochloride and / or co-crystals. [Figure 6] FIG. 6 shows IR spectra of the methanesulfonate salt and / or co-crystal of sepiapterin. [Figure 7] FIG. 7 shows IR spectra of nicotinic acid salts and / or co-crystals of sepiapterin. [Figure 8] FIG. 8 shows IR spectra of the toluenesulfonate salt and / or co-crystal of sepiapterin. [Figure 9] FIG. 9 is an IR spectrum of the co-crystal of benzenesulfonate and / or sepiapterin. [Figure 10] FIG. 10 shows IR spectra of sulfate salts and / or co-crystals of sepiapterin. [Figure 11] FIG. 11 shows IR spectra of phosphate salts and / or co-crystals of sepiapterin. [Figure 12] FIG. 12 shows IR spectra of the L-tartrate salt and / or co-crystal of sepiapterin. [Figure 13] FIG. 13 shows IR spectra of glycolate salts and / or co-crystals of sepiapterin. [Figure 14] FIG. 14 shows IR spectra of malonate salts and / or co-crystals of sepiapterin. [Figure 15] FIG. 15 is an IR spectrum of the gentisate salt and / or co-crystal of sepiapterin. [Figure 16] FIG. 16 shows IR spectra of sepiapterin fumarate salt and / or co-crystals. DETAILED DESCRIPTION OF THE INVENTION

[0138] The present invention features pharmaceutical compositions containing sepiapterin or its pharmaceutically acceptable salts and / or cocrystals, and methods for treating tetrahydrobiopterin-associated disorders using such compositions.

[0139] compound Sepiapterin The pharmaceutical compositions of the present invention comprise sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof. Sepiapterin has the following structure: JPEG2023116556000019.jpg3753

[0140] In some embodiments, sepiapterin or a pharmaceutically acceptable salt thereof and / or co-crystal described herein is present in a crystalline form in the pharmaceutical compositions of the present invention.

[0141] In some embodiments of any of the preceding compositions, the sepiapterin is in a crystalline form. In some embodiments, the sepiapterin crystalline form has at least one peak at a diffraction angle 2θ (°) of about 9.7°±0.5, about 10.2°±0.5, and / or about 11.3°±0.5, as measured by X-ray diffraction using CuKα X-ray irradiation or calculated from X-ray diffraction.

[0142] In some embodiments, the sepiapterin crystalline form is characterized by refractions at refraction angles 2θ of at least about 9.7°, about 10.2°, about 11.3°, about 14.0°, about 14.6°, about 19.9°, about 22.2°, about 25.3°, and about 32.4°. In some embodiments, the sepiapterin crystalline form is characterized by refractions at refraction angles 2θ set forth in Table 1. Alternatively, the sepiapterin, or a pharmaceutically acceptable salt and / or co-crystal thereof, exists in an amorphous form, or a combination of crystalline forms, or at least one combination of a crystalline form and an amorphous form.

[0143] In some embodiments, the pharmaceutical composition of the present invention comprises 20-30% (e.g., 20%, 22%, 25%, 27%, or 30%) sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof by total weight, or more than 20% (e.g., more than 25%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, or more than 90%) sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof by total weight.

[0144] Tetrahydrobiopterin When administered to a subject, sepiapterin is converted to tetrahydrobiopterin, which has the following structure: JPEG2023116556000020.jpg3752

[0145] Lactoylpterin An impurity that may be present in sepiapterin preparations is lactoylpterin, which may result from oxidation of sepiapterin. Lactoylpterin has the following structure: JPEG2023116556000021.jpg3755

[0146] excipients antioxidants Sepiapterin tends to oxidize rapidly when exposed to air. Therefore, the pharmaceutical composition of the present invention may contain an antioxidant. Antioxidants can minimize oxidative degradation of sepiapterin. Examples of antioxidants include 4-chloro-2,6-di-tert-butylphenol, tocopherol, α-tocopherol, alkylated diphenylamines, ascorbic acid, ascorbyl myristate, ascorbyl palmitate, ascorbyl stearate, beta-carotene, butylated hydroxyanisole, butylated hydroxytoluene, citric acid, cysteine, D-α-tocopheryl polyethylene glycol 1000 succinate, deferoxamine methanesulfonate, dodecyl gallate, ethylparaben, folic acid, fumaric acid, gallic acid, glutathione, lecithin, malic acid, methylparaben, and monothioglycol. The antioxidants include glycerol, N-acetylcysteine, nordihydroguaiaretic acid, octyl gallate, p-phenylenediamine, potassium ascorbate, potassium metabisulfite, potassium sorbate, propionic acid, propyl gallate, retinol, sorbic acid, sodium ascorbate, sodium bisulfite, sodium hydrosulfite, sodium isoascorbate, sodium metabisulfite, sodium sulfite, sodium thiosulfate, tartaric acid, tert-butylhydroquinone, tocopheryl acetate, vitamin A, vitamin B6, vitamin B12, or vitamin E. In some embodiments, the pharmaceutical compositions of the present invention include ascorbic acid, tocopherol, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, butylated hydroxytoluene, and / or butylated hydroxyanisole as antioxidants. In some embodiments, the pharmaceutical compositions of the present invention comprise ascorbic acid, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, butylated hydroxytoluene, and / or butylated hydroxyanisole as antioxidants.

[0147] In some embodiments, the pharmaceutical composition comprises less than 10% (e.g., less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%) antioxidant by total weight. In some embodiments, the pharmaceutical composition comprises 2-9% (e.g., 2-4%, 3-5%, 4-6%, 5-7%, 6-8%, or 7-9%) antioxidant by total weight. In some embodiments, the pharmaceutical composition comprises 5-100% of the USP maximum daily dose of the antioxidant, e.g., in some embodiments, the pharmaceutical composition comprises 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the USP maximum daily dose of the antioxidant. In some embodiments, the ratio of sepiapterin or its pharmaceutically acceptable salt and / or cocrystal to antioxidant is at least 1:1, e.g., 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1 wt / wt. In some embodiments, the ratio of sepiapterin or its pharmaceutically acceptable salt and / or cocrystal to antioxidant is greater than 4:1 wt / wt (e.g., greater than 5:1, greater than 6:1, greater than 7:1, greater than 8:1, greater than 9:1, greater than 10:1, greater than 15:1, or greater than 20:1). Because previous formulations of sepiapterin have contained 50% or more antioxidants (e.g., ascorbic acid), it is surprising that compositions containing less than 10% antioxidant, or even no antioxidant, are effective in stabilizing sepiapterin.

[0148] Dispersants In some embodiments, the pharmaceutical compositions of the present invention contain at least one dispersing agent. The dispersing agent may separate particles in the formulation, e.g., release its active ingredient upon contact with moisture. Examples of dispersing agents include, but are not limited to, cross-linked polyvinylpyrrolidone, carboxymethylcellulose (e.g., a croscarmellose salt, e.g., croscarmellose sodium), starch (e.g., sodium starch glycolate), or alginic acid. In some embodiments, the dispersing agent in the pharmaceutical composition is carboxymethylcellulose, such as a pharmaceutically acceptable salt of croscarmellose. In some embodiments, the pharmaceutical composition may contain 0.1 to 1.5% (e.g., 0.1%, 0.5%, 1%, or 1.5%) of dispersing agent by total weight. In some embodiments, the pharmaceutical composition contains less than 1.5% (e.g., less than 1%, less than 0.5%, or less than 0.1%) of dispersing agent.

[0149] Anti-caking agent Sepiapterin has been found to clump when added to an aqueous solution. Anti-caking agents are often added to pharmaceutical compositions, for example, to prevent the formation of lumps in the solution. Thus, in some embodiments, the pharmaceutical compositions of the present invention comprise at least one anti-caking agent. In some embodiments, the pharmaceutical compositions of the present invention comprise at least two anti-caking agents. Exemplary anti-caking agents include colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, talcum powder, sodium aluminosilicate, potassium aluminum silicate, calcium aluminosilicate, bentonite, aluminum silicate, stearic acid, and polydimethylsiloxane. In some embodiments, the at least one anti-caking agent is colloidal silicon dioxide or microcrystalline cellulose. In some embodiments, the pharmaceutical composition may comprise 65-75% (e.g., 65%, 67%, 70%, 73%, or 75%) of the anti-caking agent by total weight. In some embodiments, the pharmaceutical composition comprises both colloidal silicon dioxide and microcrystalline cellulose. In some embodiments, the pharmaceutical composition comprises 60-65% microcrystalline cellulose by total weight and 5-7% colloidal silicon dioxide by total weight.

[0150] Dosing Vehicle In some embodiments, a pharmaceutical composition of the present invention is combined with a dosing vehicle prior to administration. In some embodiments of any of the foregoing compositions, the composition may be administered in a dosing vehicle having a viscosity of, for example, about 50 to 1750 centipoise (cP) to aid in suspension and administration of the pharmaceutical composition. One type of suspending agent that can be used is a combination of glycerin and sucrose in water (e.g., MEDISCA, which contains 2.5% glycerin and 27% sucrose in water). (R)An appropriate amount of the composition can be added to the administration vehicle mixture and stirred to suspend the composition immediately prior to administration.

[0151] Other suspending agents may also be used as administration vehicles. Exemplary suspending agents include agar, alginic acid, sodium carboxymethylcellulose, carrageenan, dextrin, gelatin, guar gum, hydroxyethyl cellulose, hydroxypropyl cellulose, hypromellose, methylcellulose, polyethylene glycol, povidone, tragacanth, xanthan gum, or other suspending agents known in the art.

[0152] formulation In some embodiments, the invention features a pharmaceutical composition comprising sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof and less than 10% (e.g., 9%, 7%, 5%, 3%, 1%, 0.5%, 0.25%, or 0.1%) of an antioxidant by total weight. The antioxidant can be ascorbic acid. In some embodiments, the ratio of sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof to antioxidant is 1:1 (e.g., 2:1, 5:1, 7:1, or 10:1) wt / wt. In some embodiments, the ratio of sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof to antioxidant is greater than 4:1 wt / wt (e.g., greater than 5:1, greater than 6:1, greater than 7:1, greater than 8:1, greater than 9:1, greater than 10:1, greater than 15:1, or greater than 20:1). The pharmaceutical composition may contain 20 to 30% (e.g., 20%, 22%, 25%, 27%, or 30%) of sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof by total weight. The pharmaceutical composition may further contain a dispersing agent, such as croscarmellose sodium. The pharmaceutical composition may contain 0.1 to 1.5% (e.g., 0.1%, 0.5%, 1%, or 1.5%) of the dispersing agent by total weight. In some embodiments, the pharmaceutical composition contains at least one anti-caking agent, such as colloidal silicon dioxide or microcrystalline cellulose. The pharmaceutical composition may contain 65 to 75% (e.g., 65%, 67%, 70%, 73%, or 75%) of the anti-caking agent by total weight. In some embodiments, the pharmaceutical composition contains both colloidal silicon dioxide and microcrystalline cellulose. In some embodiments, the pharmaceutical composition comprises 60-65% by total weight of microcrystalline cellulose and 5-7% by total weight of colloidal silicon dioxide. In some embodiments, the sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof is formulated as particles of less than 140 μm (e.g., 120 μm, 110 μm, 100 μm, 90 μm, 80 μm, 70 μm, 60 μm, 50 μm, 40 μm, 30 μm, 20 μm, 10 μm, or 5 μm).In some embodiments, the pharmaceutical composition contains less than 50% (e.g., less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.3%, or less than 1%) of an impurity, e.g., lactoylpterin, e.g., the composition contains less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, or less than 0.2%.

[0153] Sepiapterin, or its pharmaceutically acceptable salts and / or cocrystals, is useful in the treatment of diseases associated with low intracellular BH4 levels, or in the treatment of diseases such as primary tetrahydrobiopterin deficiency, GTPCH deficiency, 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency, DHPR deficiency, sepiapterin reductase deficiency, dopamine-responsive dystonia, Segawa syndrome, tyrosine hydroxylase deficiency, phenylketonuria, DNAJC12 deficiency, Parkinson's disease, depression due to Parkinson's disease, impulsivity in Parkinson's disease patients, major depression, and autism. Sepiapterin in various forms, or a pharmaceutically acceptable salt thereof, of the present invention may serve as a useful treatment for diseases associated with dysfunction of various BH4-dependent metabolic pathways, including, but not limited to, cerebrovascular disease, ADHD, schizophrenia, bipolar disorder, cerebral ischemia, restless legs syndrome, obsessive-compulsive disorder, anxiety, aggression in Alzheimer's disease, cerebrovascular accident, convulsions after subarachnoid hemorrhage, myocarditis, coronary artery vasospasm, cardiac hypertrophy, arteriosclerosis, hypertension, thrombosis, infection, endotoxin shock, liver cirrhosis, hypertrophic pyloric stenosis, gastric mucosal injury, pulmonary hypertension, renal dysfunction, impotence, and hypoglycemia. Therefore, various forms of sepiapterin of the present invention, or a pharmaceutically acceptable salt thereof, can be administered to a patient in an effective amount to treat or improve a disease, disorder, or condition.

[0154] In some embodiments, the sepiapterin is a salt and / or co-crystal of sepiapterin, wherein the salt and / or co-crystal of sepiapterin is a salt and / or co-crystal with sulfuric acid, p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, malonic acid, tartaric acid (e.g., L-tartaric acid), phosphoric acid, gentisic acid, fumaric acid, glycolic acid, acetic acid, or nicotinic acid.

[0155] In some embodiments, the pharmaceutical composition comprises crystalline sepiapterin free base or a pharmaceutically acceptable salt and / or cocrystal thereof. The crystalline form of the crystalline sepiapterin free base or sepiapterin salt and / or cocrystal can occur as an anhydrate (e.g., without any bound water or solvent or hydration or solvation), or as a hydrate, partial hydrate (e.g., hemihydrate, sesquihydrate), dihydrate, or trihydrate, where the crystalline form binds water of hydration or solvent molecules associated with the crystalline form of sepiapterin or its salt. In one embodiment, the crystalline sepiapterin occurs as a monohydrate or hemihydrate.

[0156] The present invention provides pharmaceutical compositions comprising a pharmaceutically acceptable carrier and an effective amount (e.g., a therapeutically effective amount, including a prophylactically effective amount) of sepiapterin, or a salt and / or cocrystal thereof.

[0157] The pharmaceutically acceptable carrier may be any of those conventionally used, limited only by chemical and physical considerations, such as solubility and lack of reactivity with the compound, and the route of administration. Those skilled in the art will appreciate that, in addition to the pharmaceutical compositions described below, sepiapterin or its pharmaceutically acceptable salts and / or cocrystals can be formulated as a cyclodextrin inclusion complex or liposomes.

[0158] The pharmaceutically acceptable carriers described herein, for example, vehicles, adjuvants, excipients, or diluents, are well known to those skilled in the art and are readily available. Preferably, the pharmaceutically acceptable carrier is chemically inert to the active compounds and has no detrimental side effects or toxicity under the conditions of use.

[0159] dose Sepiapterin or its pharmaceutically acceptable salts and / or cocrystals can be used in any suitable dose. Appropriate doses and dosage regimens are within the skill of the art. Generally, treatment is initiated with a dose that is less than optimal. The dosage is then increased in small increments until the optimum effect under the circumstances is achieved. For convenience, the total daily dose can be divided and administered throughout the day, if desired. With appropriate doses and appropriate administration of a particular compound, the present invention provides a wide range of responses. Typically, the dosage ranges from about 2.5 to about 150 mg / kg body weight / day of the patient being treated. For example, in embodiments, sepiapterin, or a pharmaceutically acceptable salt and / or cocrystal thereof, can be administered at about 20 mg / kg to about 200 mg / kg, about 40 mg / kg to about 150 mg / kg, about 60 mg / kg to about 120 mg / kg, about 80 mg / kg to about 100 mg / kg, about 40 mg / kg to about 60 mg / kg, about 2.5 mg / kg to about 20 mg / kg, about 2.5 mg / kg to about 10 mg / kg, or about 2.5 mg / kg to about 5 mg / kg of subject body weight per day, once or more times per day, to achieve the desired therapeutic effect.

[0160] In some embodiments, the dose is sufficient to produce a level of BH4 in the CNS, e.g., as measured in the CSF, and / or is sufficient to produce a therapeutic result, e.g., an increase in the level of serotonin or dopamine in the CNS. In some embodiments, the increase in BH4 in the CNS (e.g., brain) is measured by determining the level of a metabolite of a monoamine, e.g., serotonin and / or dopamine (e.g., homovanillic acid or 5-hydroxyindoleacetic acid (5-HIAA)), in the CSF, where an increase in the metabolite in the CSF indicates an increase in BH4 levels in the CNS (e.g., brain). In some embodiments, the administered amount is sufficient to increase the level of BH4 measured in the plasma or an organ of the subject, e.g., the liver of the subject, by at least two-fold the pre-administration level of BH4.

[0161] In some embodiments, sepiapterin or its pharmaceutically acceptable salts and / or cocrystals can be formulated into unit solid oral dosage forms, such as particles. In these embodiments, each unit solid oral dosage form can contain any suitable amount of sepiapterin or its pharmaceutically acceptable salts and / or cocrystals. For example, each unit solid oral dosage form can contain about 2.5 mg, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, or about 500 mg.

[0162] Route of administration The choice of carrier can be determined in part by the particular active agent, as well as the particular method used to administer the composition. Accordingly, suitable formulations of the pharmaceutical compositions of the present invention vary widely. The following formulations for oral, aerosol, parenteral, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, intrathecal, rectal, and vaginal administration are merely exemplary and are not intended to be limiting in any way.

[0163] The pharmaceutical composition may be a liquid preparation, such as a solution, suspension, or emulsion. Formulations suitable for oral administration may consist of (a) solids or granules, such as capsules, tablets, lozenges, and troches, each containing a predetermined amount of the active ingredient; (b) powders; (c) liquids, such as an effective amount of the compound dissolved in a diluent such as water, saline, or orange juice; (d) suspensions in a suitable liquid; and (e) suitable emulsions. Solid oral dosage forms, such as capsules, tablets, and powders, are preferred. Capsule forms may be of the ordinary hard- or soft-shelled gelatin type, containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and cornstarch. Tablet forms may contain one or more of lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, coloring agents, diluents, buffers, disintegrants, wetting agents, preservatives, flavoring agents, and pharmacologically compatible carriers. Lozenge forms may contain the active ingredient in addition to a flavoring (usually sucrose and acacia or tragacanth), and similarly, pastilles may contain the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, etc., and such carriers are known in the art.

[0164] Formulations suitable for oral and / or parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions, solubilizing agents, thickeners, stabilizers, and preservatives. The compounds of the present invention may be administered in a physiologically acceptable diluent in a pharmaceutical carrier such as water, saline, aqueous dextrose and related sugar solutions, alcohols such as ethanol, benzyl alcohol, or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol and polyethylene alcohol, glycerol ketals such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers such as poly(ethylene glycol) 400, oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides with or without the addition of pharmaceutically acceptable surfactants such as soaps or detergents, suspending agents such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

[0165] Oils that can be used in parenteral formulations include petroleum, animal, vegetable, and synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, petrolatum, and mineral. Fatty acids suitable for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters. Suitable soaps for use in parenteral formulations include fatty acid alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (A) cationic detergents (e.g., dimethyldialkylammonium halides, alkylpyridinium halides, etc.), (b) anionic detergents (e.g., alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, etc.), (c) nonionic detergents (e.g., fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene-polypropylene copolymers, etc.), (d) amphoteric detergents (e.g., alkyl-β-aminopropionates, and 2-alkyl-imidazoline quaternary ammonium salts, etc.), and (3) mixtures thereof.

[0166] Parenteral formulations typically contain about 20% to about 30% by weight of sepiapterin or its pharmaceutically acceptable salts and / or cocrystals in solution. Suitable preservatives and buffers can be used in such formulations. To minimize or eliminate irritation at the injection site, such compositions may contain one or more nonionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in such compositions ranges from about 5% to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate, and high molecular weight adducts of hydrophobic bases with ethylene oxide formed by the condensation of propylene oxide with propylene glycol. Parenteral formulations can be placed in unit-dose or multi-dose hermetically sealed containers, such as ampoules and vials, and can be stored in a lyophilized (lyophilized) condition requiring only the addition of a sterile liquid carrier for injection, e.g., water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

[0167] Pharmaceutical compositions can be injectable formulations.The requirements of effective pharmaceutical carriers for injectable compositions are well known to those skilled in the art.See Remington (The Science and Practice of Pharmacy, (22nd ed.) ed. LV Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA).

[0168] Topical formulations, including those useful for transdermal drug delivery, are well known to those skilled in the art and are suitable for application to the skin in the context of the present invention. Topically applied compositions are typically in the form of liquids, creams, pastes, lotions, and gels. Topical administration includes application to oral mucosa, including the oral cavity, oral epithelium, palate, gums, and nasal mucosa. In some embodiments, the composition comprises sepiapterin or a pharmaceutically acceptable salt thereof and a suitable vehicle or carrier. It may also contain other ingredients, such as an anti-irritant. The carrier may be a liquid, solid, or semi-solid. In embodiments, the composition is an aqueous solution. Alternatively, the composition may be a dispersion of various components, an emulsion, a gel, a lotion, or a cream vehicle. In one embodiment, the primary vehicle is water or a substantially neutral or substantially neutralized biocompatible solvent. The liquid vehicle may contain other materials, such as buffers, alcohol, glycerin, mineral oil, and various emulsifiers or dispersants, as known in the art, to achieve the desired pH, consistency, and viscosity. The compositions can be prepared as solids, such as powders or granules, which can be applied directly or dissolved in water or a biocompatible solvent to form a substantially neutral or substantially neutralized solution that can be applied to the target area prior to use. In embodiments of the present invention, vehicles for topical application to the skin can include water, buffer solutions, various alcohols, glycols such as glycerin, lipid materials such as fatty acids, mineral oils, phosphoglycerides, collagen, gelatin, and silicone-based materials.

[0169] The pharmaceutical compositions may be in the form of aerosol formulations administered via inhalation. Such aerosol formulations may be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, and nitrogen. They may also be formulated as pharmaceuticals for non-pressurized preparations, such as in nebulizers or atomizers.

[0170] Further, the pharmaceutical composition may be a suppository. Formulations suitable for vaginal administration may be presented in addition to the active ingredient as pessaries, tampons, creams, gels, pastes, foams, or spray formulations as such carriers are known in the art to be appropriate.

[0171] Oral solid dosage forms Formulations for oral use include particles containing the active ingredient in a mixture with non-toxic pharmaceutically acceptable excipients, and such formulations are known to those skilled in the art (e.g., U.S. Patent Nos.: 5,817,307, 5,824,300, 5,830,456, 5,846,526, 5,882,640, 5,910,304, 6,036,949, 6,036,949, 6,372,218, which are incorporated herein by reference). Examples of excipients include inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starch including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate), granulating and disintegrating agents (e.g., cellulose derivatives including microcrystalline cellulose, starch including potato starch, croscarmellose sodium, alginates, or alginic acid), binders (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, etc.), and the like. Other pharmaceutically acceptable excipients may include colorants, flavorings, plasticizers, humectants, and buffering agents. In some embodiments, the excipients (e.g., colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, talcum powder, sodium aluminosilicate, potassium aluminum silicate, calcium aluminosilicate, bentonite, aluminum silicate, stearic acid, polydimethylsiloxane) may be present. Other pharmaceutically acceptable excipients may include colorants, flavorings, plasticizers, humectants, and buffering agents. In some embodiments, the excipients (e.g., flavorings) are packaged with the composition.In some embodiments, excipients (eg, flavorings) are packaged separately from the composition (eg, combined with the composition prior to administration).

[0172] The solid compositions of the present invention may include a coating adapted to protect the composition from undesired chemical changes (e.g., chemical degradation prior to release of the active substance). The coating may be applied to the solid dosage form in a manner similar to that described in Remington (The Science and Practice of Pharmacy, (22nd ed.) ed. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA).

[0173] Powders and granules may be prepared from the above ingredients by a conventional method, for example, using a mixer, a fluidized bed apparatus, a melt-congealing apparatus, a rotor granulator, an extrusion / spheronizing apparatus, or a spray-drying apparatus.

[0174] Treatment method The present invention is characterized by, for example, a pharmaceutical composition in an orally acceptable formulation, comprising a therapeutically effective amount of sepiapterin or a pharmaceutically acceptable salt and / or cocrystal thereof, for example, and less than 10% antioxidant. In some embodiments, the pharmaceutical composition is a granular formulation dispersed in a pharmaceutically acceptable carrier; for example, the composition can be mixed with water or another administration vehicle and ingested by a patient (e.g., within 5 to 10 minutes). Suitable formulations for use in the present invention are described in Remington (The Science and Practice of Pharmacy, (22nd ed.) ed. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA). The use of any conventional carrier in pharmaceutical compositions is contemplated unless incompatible with the active ingredient. Furthermore, for animal (e.g., human) administration, it is understood that preparations should meet sterility, thermophilicity, general safety, and purity standards as required by regulatory agencies.

[0175] The actual dosage of the compositions of the present invention administered to a patient may be determined by physical and physiological factors such as body weight, severity of the condition, type of disease being treated, previous or concurrent therapeutic interventions, the patient's idiopathies, and the route of administration. Depending on the dosage and route of administration, the preferred dosage and / or frequency of administration of an effective amount will vary depending on the subject's response. The person responsible for administration will, in any event, be able to determine the concentration of active ingredient in the composition and the appropriate dosage for the individual subject.

[0176] In some embodiments, patients receive 2.5 mg / kg / day, 5 mg / kg / day, 10 mg / kg / day, 20 mg / kg / day, 40 mg / kg / day, 60 mg / kg / day, or 80 mg / kg / day of sepiapterin, or its salts and / or cocrystals. Patients may receive a pharmaceutical composition containing sepiapterin, or its pharmaceutically acceptable salts and / or cocrystals, once daily, twice daily, or three times daily during treatment. In some embodiments, patients continue other current medications for BH4-related disorders (e.g., prescribed L-dopa / carbidopa, 5HTP, melatonin, MAO inhibitors, and dopamine receptor agonists) except for BH4 supplements (if taking BH4). Patients may not be permitted to take drugs known to inhibit folate synthesis (e.g., methotrexate, pemetrexed, trimetrexate, etc.).

[0177] In some embodiments, patients receiving BH4 therapy prior to study initiation will undergo a "washout" period during screening before receiving the pharmaceutical compositions of the present invention. Patients may be instructed to maintain a consistent diet with respect to protein and phenylalanine (Phe) intake. Dietary records may be reviewed by a qualified dietitian. Three-day total Phe levels may be calculated and recorded by the dietitian.

[0178] In some embodiments, patients taking BH4 discontinue administration of BH4 (i.e., BH4 washout). Blood samples for Phe concentration may be obtained during the BH4 washout period 7 days, 5 days, 3 days, and 1 day before treatment with the pharmaceutical compositions of the present invention, or at any time during BH4 washout until the blood Phe concentration is >360 μmol / L. In some embodiments, pre-administration blood samples are tested for sepiapteprin, Phe, BH4, and tyrosine (Tyr).

[0179] Production method of the formulation In some embodiments, pharmaceutical compositions of the present invention can be produced by mixing sepiapterin or a pharmaceutically acceptable salt and / or co-crystal thereof and an antioxidant with one or more excipients, such as a dispersing agent, and one or more anti-caking agents. In some embodiments, the components of the composition are passed through a size exclusion filter (e.g., a filter having pores of 200 μm or less) before mixing. In some embodiments, the anti-caking agent is mixed together before adding the components of the composition (e.g., sepiapterin or a pharmaceutically acceptable salt and / or co-crystal thereof, dispersing agent, and antioxidant).

[0180] In some embodiments, the pharmaceutical composition is prepared by the steps of: (a) passing at least one anti-caking agent through a size exclusion filter (e.g., a filter having pores less than 200 μm), (b) combining (e.g., by mixing in a blender) sepiacteprin, or a pharmaceutically acceptable salt and / or co-crystal thereof, an antioxidant, and optionally a dispersing agent with the at least one anti-caking agent, and (c) passing the combination from step b through a size exclusion filter (e.g., a filter having pores less than 150 μm).

[0181] In some embodiments, the at least one anti-caking agent in step a comprises two or more anti-caking agents (e.g., two anti-caking agents) mixed together before passing through the size exclusion filter. [Example]

[0182] While certain features of the invention have been illustrated and described herein, those skilled in the art will recognize many modifications, substitutions, changes, and equivalents. It is therefore to be understood that the appended claims are intended to cover all such modifications and changes that fall within the true spirit of the invention. As such, the following examples are provided to teach various aspects of the invention. These examples represent particular embodiments of aspects of the invention, and those skilled in the art will recognize that additional examples could be generated to equally teach aspects of the invention.

[0183] Example 1: Preparation of a pharmaceutical composition containing sepiapterin and an antioxidant The process for preparing a pharmaceutical composition containing sepiapterin and less than 10% antioxidant by total weight is as follows: 1 Sepiapterin (0.34 kg), ascorbic acid (0.071 kg), microcrystalline cellulose (0.85 kg), croscarmellose sodium (0.14 kg), and colloidal silicon dioxide (0.085 kg) were individually sieved through a 140 mesh screen. 2 Colloidal silicon dioxide was mixed with microcrystalline cellulose and the mixed material was passed through an 80 mesh screen. 3 The screened material from step 2 was charged into a V-blender. 4 Filtered croscarmellose, ascorbic acid, and sepiapterin were added to a V-blender. The contents of the 5 V blender were mixed for at least 10 minutes. 6 The mixture was sieved through a 140 mesh screen. 7 The mixture was stored at -20°C.

[0184] Example 2: Preparation of a pharmaceutical composition containing sepiapterin and an antioxidant The composition prepared in Example 1 (180 mg / kg sepiapterin) was added to Medisca in an amount sufficient to give a sepiapterin concentration of 58.3 mg / ml. (R) It was added to an oral mix (glycerol 2.5% (w / w), sucrose 27% (w / w) in water).

[0185] Example 3: In vivo pharmacokinetic analysis of the pharmaceutical composition of the present invention A total of 18 CD-1 mice were divided into two groups, as summarized below. Each test article was evaluated at one dose level (180 mg / kg). All animals in each group received a single oral (PO) dose of either the test article or the control product. The dose (mg / kg) was calculated based on the animal's body weight. Each group contained nine male mice. Group 1 received sepiapterin in a citrate buffer solution containing 750 mg sodium metabisulfite, 639 mg sodium citrate, 2.69 g anhydrous citric acid, and 750 mg ascorbic acid in 300 mL of sterile water. 1.5 g of carboxymethylcellulose was then added and stirred until dissolved. Sepiapterin (180 mg / kg) was added to the buffer to a concentration of 27 mg / mL. Mice in Group 1 were administered a dose of 6.67 mL / kg. Group 2 received the formulation prepared in Example 2. Mice in group 2 received a dose of 3.086 mL / kg.

[0186] All animals were weighed prior to dosing. A single oral (PO) dose was administered by oral gavage on day 1. Doses were administered on a mg / kg basis. Animals were not fasted prior to dosing.

[0187] Blood samples were collected in K2EDTA tubes and stored on wet ice protected from light. Plasma was obtained by centrifugation (35,000 rpm at 5°C) within 30 minutes of collection. Plasma samples were placed in tubes with 10% ascorbic acid in water. For every 0.1 mL of plasma, 11.1 μL of 10% ascorbic acid was added. Plasma tubes were pre-filled with 33.3 μL of 10% ascorbic acid, assuming 0.3 mL of plasma was obtained from each sample. If the plasma sample was less than 0.3 mL, the amount added was adjusted and recorded (i.e., 1 g of ascorbic acid was added for every 10 mL of water). All samples were transferred to separate 96-well plates (matrix tubes) and stored at -80°C until shipped for analysis.

[0188] Sepiapterin in mouse tissues was analyzed by reversed-phase HPLC coupled with fluorescence and electrochemical detection. The analyte in the samples was stabilized by adding ascorbic acid, dithioerythritol (DTE), and diethylenetriaminepentaacetic acid (DETAPAC). Sample preparation involved precipitation of proteins in perchloric acid containing the stabilizers, followed by collection of the clear supernatant after centrifugation.

[0189] Sepiapterin Chromatography: Sepiapterin separation was performed using a Phenomenex SphereClone 5 μm ODS(1) 250 x 4.6 mm LC column maintained at 35°C throughout the analysis. Sepiapterin was isocratically separated using 25 mM potassium phosphate buffer, pH 4.0, 17.5% methanol at a flow rate of 1.0 mL / min. Sepiapterin was detected using its native fluorescence (Ex 425 nm; Em 530 nm). The detection limit was 150 fmol on the column, with linearity maintained at at least 20 pmol.

[0190] Each sample was split into two and analyzed on consecutive days. Adjustments were made by dilution. Individual data, both raw and adjusted, and mean plasma concentrations (nM) are reported for each group and time point in Figure 1. As shown in Figure 2, the concentration of sepiapterin in plasma was greater for the formulation of Example 2 than for the sepiapterin formulation in citrate buffer.

[0191] Example 4: Measurement of BH4 levels produced by pharmaceutical compositions of the present invention Animals were administered the test articles sepiapterin or BH4 at dose levels of 20, 60, or 180 mg / kg. Blood, liver, kidney, and urine samples (when available) were collected from three animals / timepoints / groups at 0 hours (pre-dose, control only), 0.5 hours, 1 hour, 2 hours, 4 hours, and 8 hours on the day of dosing. Samples were assayed for sepiapterin or BH4 (not quantified in plasma) and subjected to noncompartmental pharmacokinetic analysis. A summary of selected pharmacokinetic parameters in each matrix is ​​shown in Figure 3.

[0192] Pharmacokinetic evaluation revealed that the maximum plasma concentration of sepiapterin was observed 30 minutes or 2 hours after administration, with a secondary absorption peak observed 8 hours after administration for the low and medium doses. Sepiapterin was detected in the liver and kidneys only 30 minutes after administration, and only in the liver for the medium and high doses. In urine, sepiapterin Cmax was observed 30 minutes after administration for the medium and high dose groups, and at the last time point for the low dose group, and was observed until the last time point for which at least one sample was obtained. Sepiapterin exposure was highest in urine among the matrices analyzed, and increased non-dose-proportionally in urine, liver, and kidney across the entire dose range. Circulating sepiapterin exposure in plasma exhibited nonlinear kinetics and was significantly greater than dose-proportional across the entire dose range, suggesting saturable plasma kinetics with increasing dose, low tissue distribution of sepiapterin, and potentially high levels of excretion via urine.

[0193] Cmax of BH4 in the liver and kidney was typically observed between 30 minutes and 2 hours after administration, and in urine between 2 and 8 hours, and was not related to dose or treatment. At certain dose levels, Cmax of BH4 in the liver was approximately 2-4 times higher with sepiapterin than with BH4 alone, and in the kidney was approximately 2-fold higher with the high dose of sepiapterin, but similar results were obtained with both treatments. Due to high individual variability and limited urine sampling, no definitive conclusions could be drawn regarding urinary BH4 exposure after both treatments. After sepiapterin or BH4 administration, BH4 exposure in the kidney, liver, and urine decreased with increasing dose, suggesting nonlinear BH4 kinetics in these matrices. The apparent elimination half-life of BH4 was 2.5 hours in the kidney at mid-dose levels after both treatments.

[0194] Example 5: In vivo evaluation of the pharmaceutical composition of the present invention in humans A Phase 1 / 2, open-label, randomized, parallel-arm, intrapatient dose-titration study can be used to evaluate the safety, pharmacokinetics, and preliminary efficacy of the formulation of Example 2 in adult and adolescent primary tetrahydrobiopterin-deficient individuals with hyperphenylalaninemia.

[0195] After consenting to the study, subjects undergo a screening test that includes medical / surgical history, demographics, vital signs, ECG, physical examination, and laboratory tests (chemistry, hematology, and urinalysis). Blood Phe levels are measured at screening and compared with the three most recent Phe levels to demonstrate that the Phe levels obtained at screening reflect the values ​​from the previous three tests. Subjects who qualify based on the screening test proceed to a BH4 washout period.

[0196] BH4 [Kuvan (R)Eligible subjects taking sapropterin dihydrochloride (sapropterin dihydrochloride) will discontinue the medication during the BH4 washout period and remain off the medication for the duration of the study. Subjects will be instructed to maintain a consistent diet (with respect to protein and Phe intake), and a 3-day food record will be collected during the BH4 washout period and throughout the study. Blood will be collected to measure Phe concentrations on days -7, -5, -3, and -1 during the BH4 washout period. Subjects will be eligible for randomization into the study if their Phe concentration is 360 μmol / L or greater at any time during the 7-day BH4 washout. Subjects whose Phe concentration does not exceed 360 μmol / L during the 7-day BH4 washout will not be eligible to participate in the study.

[0197] Subjects will be treated with sepiapterin for a total of 14 days (i.e., two 7-day treatment periods separated by a 3-4 day washout period). Subjects will be randomized into one of two cohorts, each evaluating two dose levels of the formulation of Example 2 via within-subject titration.

[0198] In Cohort 1, 2.5 mg / kg / day will be administered for 7 days in the first period, followed by a 3-4 day washout period, and then 10 mg / kg / day will be administered for 7 days in the second period (total administration days: 14 days).

[0199] In Cohort 2, 5 mg / kg / day will be administered for 7 days in the first period, followed by a 3-4 day washout period, and then 20 mg / kg / day will be administered for 7 days in the second period (total administration days: 14 days).

[0200] Subjects may receive dose escalation during Phase 2 only if they meet the criteria for intra-subject dose escalation.

[0201] During the study period, subjects will continue their other current medications for PBD (including prescribed L-dopa / carbidopa, 5HTP, melatonin, MAO inhibitors, and dopamine receptor agonists), except for BH4 supplements (if taking BH4), and will be clinically monitored according to standard of care for PBD to optimize treatment.

[0202] Safety and tolerability will be assessed primarily by adverse events (AEs), vital signs, and laboratory parameters including chemistry, hematology, urinalysis, physical examination, and 12-needle electrocardiogram (ECG). Preliminary efficacy will be assessed by reduction in plasma Phe levels. Other secondary endpoints include whole blood serotonin, serum prolactin and BH4, and urinary sepiapterin, BH4, and neotherin.

[0203] Blood samples will be collected to characterize the pharmacokinetics of sepiapterin and its effects on serum BH4, Phe, and Tyr at the following time points for each dose level: pre-dose on Day 1 (within 30 minutes of dosing), +0.5 hours (±3 minutes), +1 hour (±5 minutes), +2 hours (±6 minutes), +4 hours (±20 minutes), +6 hours (±30 minutes), +12 hours (±60 minutes, before the evening dose on Day 1), and +24 hours after the first dose of study drug (±2 hours, before the morning dose on Day 2), and on Day 7 (before the last dose of study drug).

[0204] Subjects will receive treatment with the formulation of Example 2 for a total of 14 days (i.e., two 7-day treatment periods separated by a 3-4 day washout), unless they meet criteria for stopping treatment with the formulation of Example 2.

[0205] Example 6: Preparation of salts and / or co-crystals of sepiapterin Salts and / or cocrystals of sepiapterin with hydrochloric acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, nicotinic acid, sulfuric acid, phosphoric acid, malonic acid, L-tartaric acid, fumaric acid, gentisic acid, and glycolic acid were prepared by slurrying the free base of sepiapterin and the acid in acetone / water (9 / 1, v / v) or methanol for 2 to 17 days.

[0206] The resulting salts and / or co-crystals were analyzed by DSC, TGA, HPLC, IR, and XRPD. The results are summarized in Table 19 below. The IR spectra are shown in Figures 4-16.

[0207] [Table 19]

[0208] Example 7: Stability analysis of salts and / or co-crystals of sepiapterin The stability of the prepared salts and / or co-crystals was analyzed after one week at 25°C and 60% relative humidity and at 40°C and 75% relative humidity. The results are summarized in Table 20 below. Surprisingly, of all the salts and / or co-crystals tested, the phosphate salts and / or co-crystals, the tartrate salts and / or co-crystals, and the nicotinate salts and / or co-crystals were significantly more stable than the other salts and / or co-crystals. None of the phosphate salts, tartrate salts, or nicotinate salts and / or co-crystals underwent morphological changes during the stability test, and each retained 97% or more of its purity over the two-week period tested. In fact, both the tartrate salts and nicotinate salts retained 99% or more of their purity.

[0209] [Table 20] JPEG2023116556000024.jpg86155

[0210] Example 8: Solubility and disproportionation of various sepiapterin salts and / or cocrystals The dynamic solubility of sepiapterin nicotinate, phosphate, L-tartrate, fumarate, and / or cocrystals was evaluated in water and Medisca Oral Mix. X-ray powder diffraction (XRPD) was performed on the residual solids to confirm morphological changes / disproportionation. The solids were suspended in media at a target concentration of ~7 mg / mL (calculated on a free base). The suspensions were agitated at 25 rpm in a rolling incubator for 1, 4, and 24 hours. At each time point, 1 mL of the suspension was pipetted, centrifuged at 10,000 rpm (2 minutes), and filtered through a 0.45 μm membrane to obtain the supernatant for HPLC solubility and pH testing. The residual solids were analyzed by XRPD. The solubility results are summarized in Tables 21-24.

[0211] [Table 21]

[0212] [Table 22]

[0213] [Table 23]

[0214] [Table 24]

[0215] Results: In the nicotinic acid, phosphoric acid, and L-tartaric acid samples, the residual solids converted to the free base after 1 hour in water and in Medisca Oral Mix. In the fumaric acid sample, the residual solids did not change form in water, and the crystallinity of the residual solids decreased after 1 hour in Medisca Oral Mix. Surprisingly, of the 12 different salt and / or co-crystal forms investigated, the fumarate salt and / or co-crystal was the only salt and / or co-crystal found to have high stability in the solid state stability study of Example 2 and to show no evidence of disproportionation in the disproportionation study.

[0216] Other forms While the present disclosure has been described in connection with a detailed description, it will be understood that the description is intended to be illustrative and not limiting of the scope of the present disclosure, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. A pharmaceutical composition comprising an effective amount of sepiapterin or a pharmaceutically acceptable salt thereof, and an antioxidant in an amount of less than 10% of the total weight, wherein, if the antioxidant is present, the antioxidant is ascorbic acid, an ester of ascorbic acid, or a salt of ascorbic acid.

2. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition contains lactoylpterin in an amount less than 1.3% by weight of the total amount of sepiapterin and lactoylpterin in the composition.

3. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is stable at room temperature for at least 6 months.

4. The pharmaceutical composition according to claim 1, which is formulated as a powder.

5. The pharmaceutical composition according to claim 2, which is formulated as a powder.

6. The pharmaceutical composition according to claim 3, which is formulated as a powder.

7. The pharmaceutical composition according to claim 1, wherein the composition further comprises a dispersant selected from cross-linked polyvinylpyrrolidone, carboxymethylcellulose, croscarmellose sodium, starch, or alginic acid, and the pharmaceutical composition contains 0.5 to 1.5% of the dispersant by total weight.

8. The pharmaceutical composition according to claim 7, wherein the dispersant is croscarmellose sodium, and the composition comprises 1% by weight of croscarmellose sodium.

9. The pharmaceutical composition according to claim 1, wherein the composition further comprises one or more anticaking agents selected from colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, talcum powder, sodium aluminosilicate, potassium aluminum silicate, calcium aluminosilicate, bentonite, aluminum silicate, stearic acid, and polydimethylsiloxane, and the composition comprises about 65 to 75% anticaking agent by weight.

10. The pharmaceutical composition according to claim 9, wherein the one or more anticaking agents are microcrystalline cellulose and silicon dioxide, and the composition comprises 60 to 65% by weight of microcrystalline cellulose and 5 to 7% by weight of colloidal silicon dioxide.

11. The pharmaceutical composition according to claim 1, wherein the antioxidant is selected from ascorbic acid, ascorbyl palmitate, ascorbyl myristate, ascorbyl stearate, or sodium ascorbate.

12. The pharmaceutical composition according to claim 11, wherein the antioxidant is ascorbic acid.

13. The pharmaceutical composition according to claim 1, wherein the ratio of sepiapterin to antioxidant is greater than 20:

1.

14. The pharmaceutical composition according to claim 1, wherein the composition substantially does not contain an antioxidant.

15. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition contains about 25% sepiapterin by weight.

16. The pharmaceutical composition according to claim 1, wherein the composition comprises about 25% sepiapterin, about 63% microcrystalline cellulose, about 6% colloidal silicon dioxide, about 1% croscarmellose sodium, and about 5% ascorbic acid.

17. A pharmaceutical composition according to any one of claims 1 to 16, for use in the treatment of phenylketonuria in subjects requiring it.

18. A pharmaceutical composition according to any one of claims 1 to 16, for use in reducing phenylalanine levels in subjects requiring such reduction.

19. A pharmaceutical composition according to any one of claims 1 to 16, for use in the treatment of tetrahydrobiopterin deficiency in subjects requiring it.

20. The pharmaceutical composition according to claim 19, wherein the tetrahydrobiopterin deficiency is primary tetrahydrobiopterin deficiency.

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