EA042976B1 - PHARMACEUTICALLY ACCEPTABLE SALTS OF SEPIAPTERIN - Google Patents

Description

State of the art of the present invention

Primary tetrahydrobiopterin deficiency (PBD) is caused by deficiency of GTP cyclohydrolase I (GTP-CH), 6-pyruvoyl-tetrahydropterin synthase (PTPS), or sepiapterin reductase (SR), which interfere with the biosynthesis of tetrahydrobiopterin (BH4), or by defects in BH4 recycling (pterin-4-acarbinolamine dehydratase deficiency (PCD) ) or dihydropteridine reductase (DHPR)). PBD accounts for 1 to 3% of all cases of hyperphenylalaninemia (HPA), with virtually all of the remaining cases associated with phenylalanine hydroxylase deficiency.

BH4 is an important cofactor for phenylalanine hydroxylase, tyrosine hydroxylase, tryptophan hydroxylase, glycerol fatty acid ester oxygenase, and nitric oxide synthase (NO).

In PBD, disruption of the hydroxylation of phenylalanine (Phe) to tyrosine (Tyr) results in HPA. Decreased Tyr synthesis and disruption of Tyr and tryptophan hydroxylase activity leads to decreased production of neurotransmitters and, consequently, to neuromotor deficit.

Phenotypically, BH4 deficiency appears as HPA and deficiencies in the neurotransmitter precursors, L-dopa and 5-hydroxytryptophan, and thus can be detected by screening programs that measure Phe for phenylalanine hydroxylase deficiency (with the exception of patients with SR deficiency and normal Phe concentrations).

Current treatment for BH4 deficiency consists of lowering blood Phe concentrations with oral BH4 (in GTP-CH and PTPS deficiency) and/or a low Phe diet (mainly in DHPR deficiency) and administration of neurotransmitter precursors, L-dopa and 5hydroxytryptophan. (5NTR).

There are few reports of long-term follow-up of patients with BH4 deficiency. Therapeutic strategies vary by physician and clinic, are far from clinical evidence, and include pharmacological agents, none of which have been approved in the market for this indication. Therefore, there is a need for new treatments for BH4 deficiency.

The essence of the present invention

The present invention relates to novel pharmaceutical salts of sepiapterin which exhibit improved physical properties. In particular, the present invention relates to salts of sepiapterin with improved stability, such as nicotinate, phosphate, tartrate, fumarate or hydrochloride. The present invention also relates to pharmaceutical compositions containing a pharmaceutically effective amount of one or more salts of sepiapterin, as well as methods of treating disorders associated with tetrahydrobiopterin, comprising the introduction of a sepiapterin salt of the present invention to a subject in need.

In one aspect, the present invention relates to a 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, nicotinate salt and/or co-crystal, p-toluenesulfonate salt and/or co-crystal, benzenesulfonate salt and/or co-crystal, phosphate salt and/or co-crystal ( e.g. 1:1 phosphate salt and/or co-crystal, i.e. one molecule of sepiapterin to one molecule of phosphate), malonate salt and/or co-crystal (e.g. 1:1 malonate salt and/or co-crystal, i.e. one molecule sepiapterin to one malonate molecule), tartrate salt and/or co-crystal (e.g. 1:1 tartrate salt and/or co-crystal, i.e. one sepiapterin molecule to one tartrate molecule), gentisate salt and/or co-crystal (e.g. 2: 1 gentisate salt and/or co-crystal, i.e. two molecules of sepiapterin to one molecule of gentisate), fumarate salt and/or co-crystal (e.g. 2:1 fumarate salt and/or co-crystal, i.e. two molecules of sepiapterin to one molecule fumarate), glycolate salt and/or co-crystal (e.g. 3:1 glycolate salt and/or co-crystal, i. three molecules of sepiapterin to one molecule of glycolate), acetate salt and/or co-crystal, or sulfate salt and/or co-crystal (for example, 2:1 sulfate salt and/or co-crystal, i.e. two molecules of sepiapterin to one molecule of sulfate). In some embodiments, the salt is a co-crystal with an acid. In some embodiments, the salt and/or co-crystal of sepiapterin have improved properties, such as improved stability, purity, plasma levels, and/or bioavailability.

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

In some embodiments, the salt and/or co-crystal of sepiapterin is a nicotinate salt and/or co-crystal.

In some embodiments, the salt and/or co-crystal of sepiapterin is a ptoluenesulfonate salt and/or co-crystal.

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

In some embodiments, the salt and/or co-crystal of sepiapterin is a phosphate salt and/or co-crystal, where the phosphate counterion is H ₂ PO ₄ . In some embodiments, the salt and/or co-crystal of sepiapterin is a phosphate salt and/or co-crystal, where the phosphate counterion is HPO ₄ ^2- . In some embodiments,

- 1 042976, the salt and/or co-crystal of sepiapterin is a phosphate salt and/or co-crystal, where the phosphate counterion is PO ₄ ^3- . In some embodiments, the salt and/or co-crystal of sepiapterin is a 1:1 phosphate salt and/or co-crystal, i. one molecule of sepiapterin to one molecule of phosphate. In some embodiments, the salt and/or co-crystal of sepiapterin is a 2:1 phosphate salt and/or co-crystal, i. e. two molecules of sepiapterin to one molecule of phosphate.

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

In some embodiments, the salt and/or co-crystal of sepiapterin is a 1:1 tartrate salt and/or co-crystal, i. one molecule of sepiapterin to one molecule of tartrate. In some embodiments, the salt and/or co-crystal of sepiapterin is a 2:1 tartrate salt and/or co-crystal, i. two molecules of sepiapterin to one molecule of tartrate.

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

In some embodiments, the salt and/or co-crystal of sepiapterin is a 1:1 fumarate salt and/or co-crystal, i. one molecule of sepiapterin to one molecule of fumarate. In some embodiments, the salt and/or co-crystal of sepiapterin is a 2:1 fumarate salt and/or co-crystal, i. two molecules of sepiapterin to one molecule of fumarate.

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

In some embodiments, the salt and/or co-crystal of sepiapterin is a sulfate salt and/or co-crystal, where the sulfate counterion is HSO ₄ ^- . In some embodiments, the salt and/or co-crystal of sepiapterin is a sulfate salt and/or co-crystal, where the sulfate counterion is SO ₄ ^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 to one 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. two molecules of sepiapterin to one molecule of sulfate.

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

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 (e.g., less than 30%, less than 20%, less than 10%, less than 5%, less than 1%, or 30-40 %, 25-35%, 20-30%, 15-25%, 10-20%, 5-15% or 110%) of an amorphous compound. In some embodiments, the pharmaceutically acceptable salt and/or co-crystal is substantially free of the amorphous compound. In some embodiments, the pharmaceutically acceptable salt and/or co-crystal is substantially free of any other salt or crystalline form of sepiapterin.

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal is stable. For example, in some embodiments, when exposed to 25°C and 60% relative humidity and/or 40°C and 75% relative humidity for two weeks, the pharmaceutically acceptable salt and/or co-crystal is at least 90% pure ( for example, at least 95% pure, at least 96% pure, at least 97% pure, at least 98% pure, at least 99% pure) and/or does not undergo shape change ( for example, the salt and/or co-crystal remains substantially free of free base and/or there is no change in the polymorphic form of the salt and/or co-crystal and/or free base).

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal results in increased (e.g., increased at least 5%, at least 10%, at least 15%, at least 25%, at least 50% , at least 75%, at least 95%, at least 99%, at least 100%) of a subject's plasma content (e.g., elevated plasma levels as measured by plasma levels of BH4 and/or sepiapterin, spinal fluid and/or brain of the subject) compared to other salts and/or co-crystals of sepiapterin and/or sepiapterin free base.

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal results in an increased (e.g., increased at least 5%, at least 10%, at least 15%, at least 25%, at least 50%, at least 75%, at least 95%, at least 99%, at least 100%) bioavailability in a subject compared to other salts and/or co-crystals of sepiapterin and/or sepiapterin free base .

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal has increased crystallinity compared to other salts and/or co-crystals of sepiapterin and/or sepiapterin free base.

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is the hydrochloride salt and/or co-crystal. In some embodiments, the hydrochloride salt and/or co-crystal has an endothermic transition at about 218°C (eg, 216-220°C, such as 217-219°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the hydrochloride salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), such as measured by thermogravimetric analysis.

In some embodiments, the hydrochloride salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 7.8±0.5, as measured by X-ray powder diffraction. In some embodiments, the hydrochloride salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 7.8±0.5, 12.9±0.5, and/or 26.2±0 .5 as measured by X-ray powder diffractometry. 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. 1 as measured by X-ray powder diffractometry. In some embodiments, the hydrochloride salt and/or co-crystal has all of the peaks listed in Table 1. 1 as measured by X-ray powder diffractometry.

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 transition at about 182°C (eg, 180-184°C, such as 181-183°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the methanesulfonate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the methanesulfonate salt and/or co-crystal has at least one peak at a 2θ (°) diffraction angle of 23.5±0.5 as measured by X-ray powder diffraction. In some embodiments, the methanesulfonate salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 7.9±0.5, 23.5±0.5, and/or 29.0±0 .5 as measured by X-ray powder diffractometry. In some variations

The methanesulfonate salt and/or co-crystal has one or more (for example, 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) of the peaks listed in Table. 2 as measured by X-ray powder diffractometry. In some embodiments, the methanesulfonate salt and/or co-crystal has all of the peaks listed in Table 1. 2 as measured by X-ray powder diffractometry.

Table 2. List of RPD peaks for methanesulfonate

In some embodiments, the methanesulfonate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 7.9±0.5 as measured by X-ray powder diffraction. In some embodiments, the methanesulfonate salt and/or co-crystal further has at least one peak at a 2Θ (°) diffraction angle of 7.9±0.5, 23.4±0.5, and/or 28.9±0.5 , as measured by X-ray powder diffractometry. 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. 3 as measured by X-ray powder diffractometry. In some embodiments, the methanesulfonate salt and/or co-crystal has all of the peaks listed in Table 1. 3 as measured by X-ray powder diffractometry.

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Table 3. List of RPD peaks for methanesulfonate salt

and/or co-crystal form 2 of sepiapterin 2Θ (°) Intensity 7.9 100.00 and about 21.32 12.1 22.02 13.5 79.87 15.7 11.87 17.8 9.81 19.7 10.93 21.3 26.79 23.4 96.13 24.1 24.88 24.3 22.10 25.5 9.45 26.0 11.27 27.6 7.63 28.9 95.64 31.2 4.39 36.1 6.65

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 transition at about 220°C (eg, 218-222°C, such as 219-221°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the nicotinate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the nicotinate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 24.5±0.5, as measured by X-ray powder diffraction. In some embodiments, the nicotinate salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 9.9±0.5, 23.2±0.5, and/or 24.5±0.5 , as measured by X-ray powder diffractometry. 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. 4 as measured by X-ray powder diffractometry. In some embodiments, the implementation, nicotinate salt and/or co-crystal has all the peaks listed in table. 4 as measured by X-ray powder diffractometry.

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In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is the toluenesulfonate salt. In some embodiments, the toluenesulfonate salt and/or co-crystal has an endothermic transition onset at about 190°C (e.g., 188-192°C, such as 189-191°C) and/or 263°C (e.g., 261-265° C, such as 262264°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the toluenesulfonate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the toluenesulfonate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 6.5±0.5 as measured by X-ray powder diffraction. In some embodiments, the toluenesulfonate salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 6.5±0.5, 15.1±0.5, and/or 23.4±0.5 , as measured by X-ray powder diffractometry. 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. 5 as measured by X-ray powder diffractometry. In some embodiments, the toluenesulfonate salt and/or co-crystal has all of the peaks listed in Table 1. 5 as measured by X-ray powder diffractometry.

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In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is a benzene sulfonate salt and/or co-crystal. In some embodiments, the benzenesulfonate salt and/or co-crystal has an endothermic transition onset at about 193°C (e.g., 191-195°C, such as 192-194°C) and/or 206°C (e.g., 204-208° C, such as 205-207°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the benzenesulfonate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the benzenesulfonate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 6.5±0.5 as measured by X-ray powder diffraction. In some embodiments, the benzenesulfonate salt and/or co-crystal further has at least one peak at a 2Θ (°) diffraction angle of 6.5±0.5, 14.8±0.5, and/or 19.6±0.5 , as measured by X-ray powder diffractometry. 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. 6 as measured by X-ray powder diffractometry. In some embodiments, the benzenesulfonate salt and/or co-crystal has all of the peaks listed in Table 1. 6 as measured by X-ray powder diffractometry.

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Table 6. List of RPD peaks for benzenesulfonate

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is the sulfate salt. In some embodiments, the sulfate salt and/or co-crystal has an endothermic transition at about 196°C (eg, 194198°C, such as 195-197°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the sulfate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the sulfate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 5.1±0.5 as measured by X-ray powder diffraction. In some embodiments, the sulfate salt and/or co-crystal further has at least one peak at a 2Θ (°) diffraction angle of 5.1±0.5, 7.8±0.5, and/or 23.0±0.5 , as measured by X-ray powder diffractometry. 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. 7 as measured by X-ray powder diffractometry. In some embodiments, the implementation, sulfate salt and/or co-crystal has all the peaks listed in table. 7 as measured by X-ray powder diffractometry.

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

In some embodiments, the phosphate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 25.6±0.5 as measured by X-ray powder diffraction. In some embodiments, the phosphate salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 16.6±0.5, 22.2±0.5, and/or 25.6±0.5 , as measured by X-ray powder diffractometry. 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. 8 as measured by X-ray powder diffractometry. In some embodiments, the phosphate salt and/or co-crystal has all of the peaks listed in Table 1. 8 as measured by X-ray powder diffractometry.

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is a malonate salt and/or co-crystal. In some embodiments, the malonate salt and/or co-crystal has an endothermic transition at about 175°C (eg, 173-177°C, such as 174-176°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the malonate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the malonate salt and/or co-crystal has at least one peak at a 2θ (°) diffraction angle of 6.9±0.5 as measured by X-ray powder diffraction. In some embodiments, the malonate salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 6.9±0.5, 23.8±0.5, and/or 25.5±0.5 , as measured by X-ray powder diffractometry. 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. 9 as measured by X-ray powder diffractometry. In some embodiments, the malonate salt and/or co-crystal has all of the peaks listed in Table 1. 9 as measured by X-ray powder diffractometry.

Table 9. List of RPD peaks for malonate

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is a tartrate salt (eg, L-tartrate salt). In some embodiments, the tartrate salt and/or co-crystal has an endothermic transition onset at about 156°C (e.g., 154-158°C, such as 155-157°C) and/or 175°C (e.g., 173-177° C, such as 174-176°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the tartrate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the tartrate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 7.4±0.5 as measured by X-ray powder diffraction. In some embodiments, the tartrate salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 7.4±0.5, 21.8±0.5, and/or 23.9±0.5 , as measured by X-ray powder diffractometry. 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. 10 as measured by X-ray powder diffractometry. In some embodiments, the implementation, tartrate salt and/or co-crystal has all the peaks listed in table. 10 as measured by X-ray powder diffractometry.

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Table 10. List of RPD peaks for L-tartrate

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is a fumarate salt and/or co-crystal. In some embodiments, the fumarate salt and/or co-crystal has an endothermic transition onset at about 77°C (e.g., 75-79°C, such as 76-78°C), 133°C (e.g., 131-135°C, such as 132-134°C) and/or 190°C (eg 188-192°C, such as 189-191°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the fumarate salt and/or co-crystal has a weight loss at 31-150°C of less than 5% (e.g., less than 4%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the fumarate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 24.0±0.5, as measured by X-ray powder diffraction. In some embodiments, the fumarate salt and/or co-crystal further has at least one peak at a 2θ diffraction angle (°) of 11.4±0.5, 11.9±0.5, and/or 24.0±0.5 , as measured by X-ray powder diffractometry. 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. 11 as measured by X-ray powder diffractometry. In some embodiments, the fumarate salt and/or co-crystal has all of the peaks listed in Table 1. 11 as measured by X-ray powder diffractometry.

Table 11. List of RPD peaks for fumarate

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In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is a gentisate salt and/or co-crystal. In some embodiments, the gentisate salt and/or co-crystal has an endothermic transition onset at about 83°C (e.g., 81-85°C, such as 82-84°C), 134°C (e.g., 132-136°C, such as 133-135°C) and/or 149°C (eg 147-151°C such as 148-150°C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the gentisate salt and/or co-crystal has a weight loss at 31-150°C of less than 7% (e.g., less than 5%, less than 3%, less than 2%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the gentisate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 7.1±0.5 as measured by X-ray powder diffraction. In some embodiments, the gentisate salt and/or co-crystal further has at least one peak at a 2θ (°) diffraction angle of 7.1±0.5, 8.7±0.5, and/or 26.7±0.5 , as measured by X-ray powder diffractometry. 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. 12 as measured by X-ray powder diffractometry. In some embodiments, the gentisate salt and/or co-crystal has all of the peaks listed in Table 1. 12 as measured by X-ray powder diffractometry.

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In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is a glycolate salt and/or co-crystal. In some embodiments, the glycolate salt and/or co-crystal has an endothermic transition onset at about 79°C (e.g., 77-81°C, such as 78-80°C), 90°C (e.g., 88-92°C, such as 89-91°C), 132°C (for example, 130-134°C, such as 131-133°C) and/or 152°C (for example, 150-154°C, such as 151-153° C) on a differential scanning calorimetry (DSC) graph. In some embodiments, the glycolate salt and/or co-crystal has a weight loss at 31-150°C of less than 21% (e.g., less than 15%, less than 10%, less than 5%, or less than 1%), as measured by thermogravimetric analysis.

In some embodiments, the glycolate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 7.6±0.5 as measured by X-ray powder diffraction. In some embodiments, the glycolate salt and/or co-crystal further has at least one peak at a 2Θ (°) diffraction angle of 7.6±0.5, 10.7±0.5, and/or 24.0±0.5 , as measured by X-ray powder diffractometry. 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. 13 as measured by X-ray powder diffractometry. In some embodiments, the implementation, glycolate salt and/or co-crystal has all the peaks listed in table. 13 as measured by X-ray powder diffractometry.

- 13 042976

Table 13. List of RPD peaks for glycolate

In some embodiments, the acetate salt and/or co-crystal has at least one peak at a 2Θ (°) diffraction angle of 6.2±0.5 as measured by X-ray powder diffraction. In some embodiments, the acetate salt and/or co-crystal further has at least one peak at a 2Θ (°) diffraction angle of 6.2±0.5, 12.0±0.5, and/or 18.1±0.5 , as measured by X-ray powder diffractometry. In some embodiments, the acetate 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 X-ray powder diffractometry. In some embodiments, the acetate salt and/or co-crystal has all of the peaks listed in Table 1. 14 as measured by X-ray powder diffractometry.

In another aspect, the present invention relates to a composition (eg, solid composition) containing any of the above pharmaceutically acceptable salts and/or co-crystals and a pharmaceutically acceptable excipient. In some embodiments, the composition is stable at room temperature for at least 6 months. For example, the level of sepiapterin or a salt and/or co-crystal thereof in the composition decreases by less than 5% (e.g., less than 4%, less than 3%, less than 2%, less than 1%) when the composition is stored at room temperature for 6 months 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%) when the composition is stored at room temperature for 6 months. In some embodiments, the pharmaceutically acceptable salt and/or co-crystal contains less than 10% by weight (eg, less than 5% or less than 1%) of the amorphous compound. In some embodiments, the pharmaceutically acceptable salt and/or co-crystal is substantially free of the amorphous compound.

In some embodiments of any of the above compositions, the composition comprises an antioxidant (e.g., ascorbic acid) wherein the ratio of pharmaceutically acceptable salt and/or co-crystal of sepiapterin to antioxidant is greater than 4:1 (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) by weight (e.g., weight of salt and/or co-crystal to an antioxidant).

In some embodiments of any of the above compositions, the composition contains an antioxidant, where the pharmaceutical composition contains more pharmaceutically acceptable salt of sepiapterin than the antioxidant by weight. For example, in some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin 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) by weight.

In some embodiments of any of the above compositions, the composition comprises an antioxidant, wherein the composition contains 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 (eg, ascorbic acid) relative to the total weight. In some embodiments, the pharmaceutical composition is substantially free of an antioxidant.

In some embodiments of any of the above compositions, the composition contains less than 10% (e.g., less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less 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%) lactoylpterin relative to the total weight. In some embodiments, the pharmaceutical composition contains less than 1.3% lactoylpterin. In some embodiments, the pharmaceutical composition further comprises an antioxidant (eg, ascorbic acid). In other embodiments, the pharmaceutical composition does not contain an antioxidant.

In some embodiments of any of the above compositions, the composition is formulated for use as a suspension.

In some embodiments of any of the above pharmaceutical compositions, the antioxidant is 4-chloro-2,6-di-tert-butylphenol, tocopherol, alpha-tocopherol, alkylated diphenylamines, ascorbic acid, ascorbyl myristate, ascorbyl palmitate, ascorbyl stearate, beta-carotene, butylated hydroxyanisole, butylated hydroxytoluene, citric acid, cysteine, D-alpha-tocopheryl polyethylene glycol 1000 succinate, deferoxamine mesylate, dodecyl gallate, ethylparaben, folic acid, fumaric acid, gallic acid, glutathione, lecithin, malic acid, methylparaben, monothioglycerin, 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-butyl hydroquinone, tocopheryl acetate, vitamin A, vitamin B6, vitamin B12 or vitamin E, or a combination thereof. In some embodiments, any of the above pharmaceutical compositions, the antioxidant is ascorbic acid, tocopherol, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, butylated hydroxytoluene, and/or butylated hydroxyanisole.

In some embodiments of any of the above compositions, the pharmaceutical composition contains about 20-95% (e.g., about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, approximately 28%, approximately 29%, approximately 30%, approximately 40%, approximately 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, approximately 95% or approximately 20-30%, approximately 25-45%, about 40-60%, about 50-75%, about 70-90%, about 85-95%) of sepiapterin or a pharmaceutically acceptable salt and/or co-crystal thereof, based on total weight.

In some embodiments of any of the above compositions, the pharmaceutical composition further comprises a dispersant (eg, carboxymethyl cellulose 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 contains 0.1-1.5% (e.g., 0.1-0.3%, 0.2-0.4%, 0.3-0.5% , 0.4-0.6%, 0.5-0.7%, 0.6-0.8%, 0.7-0.9%, 0.8-1%,

- 15 042976

0.9-1.1%, 1-1.2%, 1.1-1.3%, 1.2-1.4%, or 1.3-1.5%) dispersant (e.g. croscarmellose sodium ) relative to the total weight.

In some embodiments of any of the above compositions, the pharmaceutical composition contains at least one anti-caking agent or bulking agent (eg, bulking agent and anti-caking agent). In some embodiments, at least one anti-caking agent or bulking agent is colloidal silicon dioxide or microcrystalline cellulose. In some embodiments, the pharmaceutical composition contains 60-80% (e.g., about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68% , approximately 69%, approximately 70%, approximately 71%, approximately 72%, approximately 73%, approximately 74%, approximately 75%, approximately 76%, approximately 77%, approximately 78%, approximately 79%, or approximately 80%) anti-caking agent and/or bulking agent relative to the total weight. In some embodiments of any of the above compositions, the pharmaceutical composition contains both colloidal silicon dioxide and microcrystalline cellulose. In some embodiments of any of the above compositions, the pharmaceutical composition contains 60-65% (about 60%, about 61%, about 62%, about 63%, about 64%, or about 65%) microcrystalline cellulose based on total weight and 2 -15% (e.g., 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 silica, based on total weight.

In some embodiments of any of the above compositions, sepiapterin is formulated as particles with a particle size 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).

In some embodiments of any of the above compositions, the pharmaceutical composition is formulated into particles (eg, particles for use in suspension). In some embodiments, the particles are less than 200 microns in size (eg, less than 180 microns, less than 160 microns, less than 140 microns, less than 120 microns, less than 100 microns, or less than 80 microns).

In some embodiments of any of the above compositions, the pharmaceutical composition contains less than 50% (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 %) lactoylpterin by weight of the combined amount of sepiapterin or salt and lactoylpterin. In some embodiments, the pharmaceutical composition contains less than 1.3% lactoylpterin.

In some embodiments of any of the above compositions, the pharmaceutical composition further comprises a medium (eg, a medium with a viscosity of approximately 50-1750 centipoise).

In some embodiments of any of the above compositions, the pharmaceutical composition, upon administration to a subject, results in high levels in plasma or liver cells (e.g., at least 1.1 fold, at least 1.2 fold, at least 1.3 multiples, at least 1.4 folds, at least 1.5 folds, at least 2 folds, at least 3 folds, or at least 4 folds) of tetrahydrobiopterin (e.g., as measured by T _max , C max , AUC, or concentration in plasma 15 minutes after administration) compared to what is obtained by administering a pharmaceutical composition containing an equivalent dose of tetrahydrobiopterin.

In some embodiments of any of the above compositions, the pharmaceutical composition, upon administration to a subject, results in high levels in plasma or liver cells (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) tetrahydrobiopterin (e.g., as measured by T _max , C _max , AUC or plasma concentration 15 minutes after administration) compared to that obtained by administering a pharmaceutical composition containing an equivalent dose of sepiapterin or a pharmaceutically acceptable salt thereof and more than 10% of an antioxidant.

In one aspect, the present invention relates to a method for preparing a pharmaceutical composition containing a pharmaceutically acceptable salt and/or co-crystal of sepiapterin. This method includes: a) mixing microcrystalline cellulose and colloidal silicon dioxide; b) adding a pharmaceutically acceptable salt and/or co-crystal of sepiapterin, a dispersant and an antioxidant to the mixture of step a; and c) mixing microcrystalline cellulose, colloidal silicon dioxide,

- 16 042976 pharmaceutically acceptable salt and/or co-crystal of sepiapterin, dispersant and antioxidant, thereby obtaining a pharmaceutical composition containing a pharmaceutically acceptable salt and/or co-crystal of sepiapterin.

In some embodiments, a mixture of microcrystalline cellulose and colloidal silica is passed through a filter with pores 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) before step b.

In some embodiments, a mixture of microcrystalline cellulose, colloidal silica, sepiapterin, dispersant, and antioxidant is passed through a filter with pores smaller 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).

In some embodiments of any of the above compositions, the percentage by weight is measured on a dry composition (eg, prior to being suspended in a liquid such as water).

In some embodiments of any of the above methods, the antioxidant is ascorbic acid. In some embodiments of the above methods, the dispersant is croscarmellose sodium.

In one aspect, the present invention relates to a method of treating a tetrahydrobiopterin-associated disease (e.g., phenylketonuria or tetrahydrobiopterin deficiency) in a subject in need thereof, the method comprising administering an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention provides a method for increasing tetrahydrobiopterin levels in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method for lowering phenylalanine levels in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention provides a method for increasing phenylalanine hydroxylase activity in a subject, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method of treating phenylketonuria in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method of treating gastroporesis in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method for increasing serotonin levels in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention provides a method for increasing tryptophan hydroxylase activity in a subject, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method for increasing dopamine levels in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method for increasing tyrosine hydroxylase activity in a subject, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method for increasing nitric oxide synthase activity in a subject, the method comprising administering to the subject an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method for increasing the activity of an alkylglycerol monooxygenase in a subject, the method comprising administering to the subject an effective amount

- 17 042976 any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions.

In one aspect, the present invention relates to a method for increasing the level of homovanylic acid and/or 5-hydroxyindoleacetic acid in a subject, the method comprising administering an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions. In one embodiment, the pharmaceutically acceptable salt and/or co-crystal is methanesulfonate salt and/or co-crystal, nicotinate salt and/or co-crystal, toluenesulfonate salt and/or co-crystal, benzene sulfonate salt and/or co-crystal, sulfate salt and/or co-crystal, phosphate salt and/or co-crystal, malonate salt and/or co-crystal, tartrate salt and/or co-crystal, fumarate salt and/or co-crystal, gentisate salt and/or co-crystal, or glycolate salt and/or co-crystal. In some embodiments, the level of homovanilic acid and/or 5-hydroxyindoleacetic acid in the cerebrospinal fluid (CSF) of a subject is increased. In some embodiments, the level of homovanylic acid and/or 5-hydroxyindoleacetic acid in the subject (e.g., in the CSF of the subject) is increased by at least 5% compared to the pre-administration concentration (e.g., the level is increased by at least 10%, according to at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least at least 90%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300% compared to pre-administration levels).

In some embodiments, prior to administration of a pharmaceutically acceptable salt and/or co-crystal or pharmaceutical composition, the subject has levels of homovanylic acid and/or 5hydroxyindoleacetic acid that are less than 50% (e.g., less than 40%, less than 30%) of the levels an average subject (eg, the subject has CSF levels of homovanylic acid less than 15 ng/ml and/or has CSF levels of 5-hydroxyindoleacetic acid less than 5 ng/ml). In some embodiments, the subject has not been diagnosed with a BH4-associated disorder. In some embodiments, the subject has no symptoms of a disorder associated with BH4. In some embodiments, following administration of a pharmaceutically acceptable salt and/or co-crystal or pharmaceutical composition, the levels of homovanylic acid and/or 5hydroxyindoleacetic acid in the subject are greater than 50% of the levels of the average subject (e.g., the subject has levels of homovanylic acid in the CSF greater than 15 ng/mL and/or has CSF levels of 5-hydroxyindoleacetic acid greater than 5 ng/mL).

In some embodiments of the above methods, an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions comprises an amount sufficient to increase the plasma level of tetrahydrobiopterin in a subject 1 hour after administration by at least two times (for example, at least 3, 4, 5, 6, 7, 9 or 10 times) compared with the level of tetrahydrobiopterin before administration.

In some embodiments of the above methods, an effective amount of any of the above pharmaceutically acceptable salts and/or co-crystals or pharmaceutical compositions comprises an amount sufficient to increase the level of tetrahydrobiopterin in the CSF and/or brain of the subject 1 hour after administration, by at least two times (for example, at least 3, 4, 5, 6, 7, 9 or 10 times) compared with the level of tetrahydrobiopterin before administration.

In some embodiments of the above methods, the subject is a human. In some embodiments of the above methods, the method includes mixing any of the above pharmaceutically acceptable salts or pharmaceutical compositions with a medium prior to administration.

Definitions.

In this application, unless otherwise clear from the context, (i) the term a should be understood to mean at least one; (ii) the term or can be understood to mean and/or; (iii) the terms comprising and comprising are to be understood as including detailed components or steps, whether presented alone or together with one or more additional components or steps; and (iv) the term approximately is to be understood as allowing for standard variations, as will be understood by those skilled in the art; and (v) where ranges are specified, endpoints are included.

As used in the present invention, the term introduction refers to the introduction of the composition to the subject. Administration to an animal (eg, human) subject may be by any suitable route. For example, in some embodiments, administration may be bronchial (including bronchial instillation), buccal, enteral, intradermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal , subcutaneous, sublingual, topical, tracheal (including intratracheal instillation), transdermal, vaginal or vitreal.

- 18 042976

The term anti-caking agent refers to an additive added to powdered or granular pharmaceutical compositions to prevent clumping. 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, powdered talc, sodium aluminosilicate, potassium aluminosilicate, calcium aluminosilicate, bentonite, aluminosilicate, stearic acid and polydimethylsiloxane.

The term antioxidant refers to agents that minimize 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-alphatocopheryl polyethylene glycol 1000 succinate, deferoxamine methanesulfonate, dodecyl gallate, ethyl paraben, folic acid, fumaric acid, gallic acid, lecithin, malic acid, methyl paraben, monothioglycerin, nordihydroguaiaretin 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-butyl hydroquinone, tocopheryl acetate, vitamin A, vitamin B6 and vitamin B12.

As used herein, the term BH4-associated disorder or tetrahydrobiopterin-associated disorder refers to any disease or disorder that can benefit from modulation (eg, elevation) of BH4 levels, such as phenylketonuria.

By determining the connection level is meant the detection of a connection by methods known in the art, either directly or indirectly. Direct definition refers to performing a process (eg, performing an analysis or test on a sample, or analyzing a sample as the term is defined herein) to obtain a physical object or value. An indirect definition refers to the receipt of a physical object or value from another party or source (for example, a third-party laboratory that directly receives a physical object or value). Methods for measuring the amount of a compound typically include, but are not limited to, liquid chromatography (LC)-mass spectrometry.

The term dispersant refers to an agent used in pharmaceutical formulations that causes the particles in the formulation to separate, for example, releasing their drugs upon contact with moisture. Examples include cross-linked polyvinylpyrrolidone, carboxymethylcellulose (eg a salt of croscarmellose, eg croscarmellose sodium), starch (eg sodium starch glycolate) or alginic acid.

The effective amount of a compound may vary depending on factors such as disease state, age, sex, weight of the individual, and the ability of the compound to elicit the desired response. A therapeutically effective amount includes an amount in which any toxic or deleterious effects of the compound are outweighed by the therapeutic benefits. A therapeutically effective amount also includes an amount sufficient to provide a benefit, such as a clinical benefit.

By increasing the activity of an enzyme is meant an increase in the level of activity associated with the enzyme, for example, phenylalanine hydroxylase, or an associated subsequent effect. A non-limiting example of an increase in enzyme activity includes an increase in phenylalanine hydroxylase activity, resulting in a decrease in phenylalanine levels. The level of enzyme activity can be measured using any method known in the art.

By level is meant the connection level compared to the reference. The reference may be any suitable reference as defined in the present invention. Under reduced or increased levels of compounds refers to a decrease or increase in the level of compounds compared to the reference (for example, decrease or increase of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35 %, approximately 40%, approximately 45%, approximately 50%, approximately 55%, approximately 60%, approximately 65%, approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500% or more; decrease or increase by more than about 10%, about 15%, about 20%, about 50%, about 75 %, about 100%, or about 200%, compared to the reference; decrease or increase less than about 0.01 times, about 0.02 times, about 0.1 times, about 0.3 times, about 0.5 times , about 0.8 times or less; or more than about 1.2 times, about 1.4 times, about 1.5 times, about 1.8 times, about 2.0 times, about 3.0 times, about 3.5 times , about 4.5 times, about 5.0 times, about 10 times, about 15 times, about 20 times, about 30 times, about 40 times, about 50 times, about 100 times, about 1000 times or more). The level of a compound may be expressed in mass/volume (eg, g/dL, mg/mL, μg/mL, ng/mL) or as a percentage of the total compound in the sample.

The term pharmaceutical composition, as used in the present invention, is a composition containing a compound described in the present invention, formulated with a pharmaceutically acceptable excipient. The pharmaceutical composition may be a pharmaceutical composition prepared or marketed with government regulatory approval as part of a therapeutic regimen for the treatment of a disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (eg tablets, capsules, caplet, gelatin capsule, suspension, solution or syrup); for topical administration (for example, in the form of a cream, gel, lotion or ointment); for intravenous administration (for example, in the form of a sterile solution, free from particles of emboli and in a solvent system suitable for intravenous use); or in the form of any other pharmaceutically acceptable composition.

As used in the present invention, the term pharmaceutically acceptable salt means any pharmaceutically acceptable salt of sepiapterin. Pharmaceutically acceptable salts include sepiapterin pair ions in the solid state and/or in solution. A pharmaceutically acceptable co-crystal contains sepiapterin in the form of a free base and an acid in the solid state. A mixture of a salt form and a co-crystal form may be present in one composition. For example, pharmaceutically acceptable salts of sepiapterin include salts which, under reasonable medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic response, and a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Remington (Remington: The Science and Practice of Pharmacy, (22nd ed.) ed. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA). Salts can be obtained in situ during the final isolation and purification of the compounds described in the present invention, or separately by the reaction of the free base group with a suitable organic acid.

Sepiapterin can be obtained in the form of 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 the corresponding salts are well known in the art.

By benchmark is meant any suitable benchmark used to compare connection levels. The reference can be any sample, standard, standard curve or level used for comparison. The reference can be a reference standard or an analytical standard or level. The control sample can be, for example, a control, for example, a predetermined value of a negative control, such as a normal control or a previous sample taken from the same subject; a sample from a normal healthy subject, such as a normal cell or normal tissue; a sample (eg, cell or tissue) from a subject not suffering from a disease; a sample from a subject who has been diagnosed with a disease but has not yet been treated with a compound of the present invention; a sample from a subject who has been treated with a compound of the present invention; or a sample of a purified compound (eg, any of those described herein) at a known normal level. By analytical standard or level is meant the value or number obtained from a reference sample. The normal reference value is a predetermined value indicating the absence of disease, such as the value expected in a healthy control subject. Normally, a normal reference value is expressed as a range (between X and Y), an upper threshold (not higher than X), or a lower threshold (not lower than X). A subject having a measured value within the normal reference range for a particular biomarker is generally referred to as within normal range for that biomarker. A normal analytical standard or level may be a value or number obtained from a normal subject not suffering from a disease or disorder (eg, cancer); a subject treated with a compound of the present invention. In preferred embodiments, the reference sample, standard, or level is compared to the test sample on at least one of the following criteria: age, weight, sex, disease stage, and general health. A standard curve of levels of a purified compound, such as any compound described herein, within the normal reference range may also be used as a reference.

As used in the present invention, the term subject or patient refers to any organism into which a composition according to the present invention may 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,

- 20 042976 non-human primates and humans). The subject may be seeking or in need of treatment, require treatment, receive treatment, receive treatment in the future, or be a human or animal under the care of a qualified specialist for a particular disease or condition.

As used in the present invention, the terms treat or treatment refer to both therapeutic treatment and prophylactic or preventive measures, the purpose of which is to prevent or slow down (reduce) an undesirable physiological condition, disorder or disease, or to obtain favorable or desired clinical results. Positive or desired clinical results include, but are not limited to, symptomatic relief; reducing the severity of the condition, disorder or disease; a stabilized (ie not worsening) condition, disorder or disease; delaying the onset or slowing the progression of the condition, disorder or disease; improvement in the condition, disorder or disease state or remission (partial or complete), detectable or undetectable; improvement in at least one measurable physical parameter, not necessarily perceptible by the patient; or an improvement or improvement in a condition, disorder, or disease. Treatment involves obtaining a clinically significant response without undue side effects. Treatment also includes prolonging survival compared to expected survival without treatment.

Unless otherwise indicated, all technical and scientific terms used in the present invention have the same meaning as generally understood by a person skilled in the art to which the present invention belongs. The present invention describes methods and materials for use in the present invention; other suitable methods and materials known in the art may 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 in the present invention are incorporated by reference in their entirety. In the event of a conflict, the present invention, including definitions, will govern.

The details of one or more embodiments of the present invention are set forth in the description below. Other features, objects and advantages of the present invention will be apparent from the description and claims.

Brief description of the drawings

Fig. 1 is the IR spectrum of the free base of sepiapterin.

Fig. 2 is the IR spectrum of the hydrochloride salt and/or co-crystal of sepiapterin.

Fig. 3 is the IR spectrum of the methanesulfonate salt and/or co-crystal of sepiapterin.

Fig. 4 is an IR spectrum of a nicotinate salt and/or a co-crystal of sepiapterin.

Fig. 5 is an IR spectrum of a toluenesulfonate salt and/or a co-crystal of sepiapterin.

Fig. 6 is an IR spectrum of a benzenesulfonate salt and/or a co-crystal of sepiapterin.

Fig. 7 is the IR spectrum of the sulfate salt and/or co-crystal of sepiapterin.

Fig. 8 is an IR spectrum of a phosphate salt and/or a co-crystal of sepiapterin.

Fig. 9 is an IR spectrum of an L-tartrate salt and/or a co-crystal of sepiapterin.

Fig. 10 is an IR spectrum of a glycolate salt and/or a co-crystal of sepiapterin.

Fig. 11 is an IR spectrum of a malonate salt and/or a co-crystal of sepiapterin.

Fig. 12 is an IR spectrum of a gentisate salt and/or a co-crystal of sepiapterin.

Fig. 13 is an IR spectrum of a fumarate salt and/or a co-crystal of sepiapterin.

Detailed description

The present invention relates to pharmaceutically acceptable salts and/or co-crystals of sepiapterin, their compositions and methods of treatment of disorders associated with tetrahydrobiopterin, these compositions. The inventors of the present invention have unexpectedly found that certain salts and/or co-crystals of sepiapterin have improved properties, such as increased stability, purity, plasma concentration and/or bioavailability.

Compound.

Sepiapterin.

The pharmaceutical compositions of the present invention contain sepiapterin or a pharmaceutically acceptable salt and/or co-crystal thereof. Sepiapterin has the structure

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is present in the pharmaceutical composition of the present invention in crystalline form as described herein.

In some embodiments, the pharmaceutical composition of the present invention contains 20-30% of a pharmaceutically acceptable salt and/or co-crystal of sepiapterin based on total weight, such as 20%, 22%, 25%, 27%, or 30%. In some embodiments, the pharmaceutical composition contains more than 20% pharmaceutically acceptable salt and/or co-crystal of sepiapterin based on total weight, 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%.

T tetrahydrobiopterin.

Sepiapterin, when administered to a subject, is converted to tetrahydrobiopterin. Tetrahydrobiopterin has the structure

Lactoilpterin.

An impurity that is present in sepiapterin preparations is lactoylpterin, which may result from the oxidation of sepiapterin. Lactoylpterin has the structure

Excipients.

Antioxidants.

Sepiapterin is prone to rapid oxidation in air. Accordingly, the pharmaceutical composition of the present invention may contain an antioxidant.

The antioxidant can minimize the oxidative degradation of sepiapterin. Examples of antioxidants include, but are not limited to, 4-chloro-2,6-di-tert-butylphenol, tocopherol, alpha-tocopherol, alkylated diphenylamines, ascorbic acid, ascorbyl myristate, ascorbyl palmitate, ascorbyl stearate, beta-carotene, butylated hydroxyanisole, butylated hydroxytoluene, citric acid , cysteine, D-alpha-tocopheryl polyethylene glycol 1000 succinate, deferoxamine methanesulfonate, dodecylgallate, ethylparaben, folic acid, fumaric acid, gallic acid, glutathione, lecithin, malic acid, methylparaben, monothioglycerin, N-acetylcysteine, nordihydroguaiaretic acid, ok tilgallate, p- phenylenediamine, potassium ascorbate, potassium metabisulfite, potassium sorbate, propynic acid, propyl gallate, retinol, sorbic acid, sodium ascorbate, sodium bisulfite, sodium hydrosulfite, sodium isoascorbate, sodium metabisulfite, sodium sulfite, sodium thiosulfate, tartaric acid, tert-butyl hydroquinone, tocophenylacetate , vitamin A, vitamin B6, vitamin B12, or vitamin E. In some embodiments, the pharmaceutical composition of the present invention contains ascorbic acid, tocopherol, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, butylated hydroxytoluene, and/or butylated hydroxyanisole as an antioxidant.

In some embodiments, the pharmaceutical composition contains less than 10% antioxidant by weight, such as 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%. In some embodiments, the pharmaceutical composition contains 2-9% antioxidant based on total weight, such as 2-4%, 3-5%, 4-6%, 5-7%, 6-8%, or 7-9%. In some embodiments, the pharmaceutical composition contains 5-100% of the maximum daily dose of USP antioxidant, for example, in some embodiments, the pharmaceutical composition contains 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90 or 100% of the maximum daily dose of USP antioxidant. In some embodiments, the ratio of sepiapterin or a pharmaceutically acceptable salt and/or co-crystal thereof to antioxidant is at least 1:1, such as 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1 w/w. In some embodiments of any of the above compositions, the composition comprises an antioxidant (e.g., ascorbic acid) wherein the ratio of a pharmaceutically acceptable salt and/or co-crystal of sepiapterin to the antioxidant is greater than 4:1 (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) by weight (eg, weight of salt to antioxidant).

Because previous formulations of sepiapterin have contained up to 50% antioxidant (eg, ascorbic acid) or more, it is surprising that compositions containing less than 10% antioxidant or even no antioxidant were effective in stabilizing pharmaceutically acceptable salts and/or co-crystals of sepiapterin.

Dispersants.

In some embodiments, the pharmaceutical composition of the present invention contains at least one dispersant. The dispersant may cause the particles in the formulation to separate, for example, releasing its medicinal substances upon contact with moisture. Examples of a dispersant include, but are not limited to, cross-linked polyvinylpyrrolidone, carboxymethyl cellulose (e.g.,

- 22 042976 croscarmellose salt, eg croscarmellose sodium), starch (eg sodium starch glycolate) or alginic acid. In some embodiments, the dispersant in the pharmaceutical composition is a carboxymethyl cellulose, such as a pharmaceutically acceptable salt of croscarmellose. In some embodiments, the pharmaceutical composition may contain 0.1-1.5% dispersant based on total weight, such as 0.1%, 0.5%, 1%, or 1.5%. In some embodiments, the pharmaceutical composition contains less than 1.5% dispersant, such as less than 1%, less than 0.5%, or less than 0.1%.

Anti-clotting agents.

Sepiapterin has been found to form lumps when added to aqueous solutions. Anti-caking agents are often added to pharmaceutical compositions to prevent clumping, for example in solutions. Accordingly, in some embodiments, the pharmaceutical compositions of the present invention contain at least one anti-caking agent. In some embodiments, the pharmaceutical compositions contain 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, talc powder, sodium aluminum silicate, potassium aluminum silicate, calcium aluminum silicate, bentonite, aluminum silicate, stearic acid and polydimethylsiloxane. In some embodiments, at least one anti-caking agent is colloidal silicon dioxide or microcrystalline cellulose. In some embodiments, the pharmaceutical composition may contain 65-75% anti-caking agent based on total weight, such as 65%, 67%, 70%, 73%, or 75%. In some embodiments, the pharmaceutical composition contains both colloidal silicon dioxide and microcrystalline cellulose. In some embodiments, the pharmaceutical composition contains 60-65% microcrystalline cellulose based on total weight and 5-7% colloidal silicon dioxide based on total weight.

Carrier.

In some embodiments, the pharmaceutical composition of the present invention is mixed with a carrier prior to administration. In some embodiments of any of the above compositions, the composition may be administered in a vehicle with a viscosity of about 50-1750 centipoise (cP), for example, to aid in suspending and dosing the pharmaceutical composition. One type of suspending agent that can be used is a combination of glycerol and sucrose in water (eg MEDISCA® oral mixture with 2.5% glycerol and 27% sucrose in water). An appropriate amount of the composition may be added to the carrier mixture and shaken to suspend the composition immediately prior to administration.

Other suspending agents may also be used as a carrier. Examples of suspending agents include agar, alginic acid, sodium carboxymethylcellulose, carrageenan, dextrin, gelatin, guar gum, hydroxyethylcellulose, hydroxypropylcellulose, hypromellose, methylcellulose, polyethylene glycol, povidone, tragacanth, xanthan gum, and other suspending agents known in the art.

Compositions.

In some embodiments, the present invention relates to a pharmaceutical composition containing a pharmaceutically acceptable salt and/or co-crystal of sepiapterin, for example, and less than 10% relative to the total weight of the antioxidant, for example, 9, 7, 5, 3, 1, 0.5, 0.25 or 0.1%. The antioxidant may be ascorbic acid. In some embodiments, the ratio of pharmaceutically acceptable salt and/or co-crystal of sepiapterin to antioxidant is 1:1, such as 2:1, 5:1, 7:1, or 10:1 w/w. In some embodiments of any of the above compositions, the composition comprises an antioxidant (e.g., ascorbic acid) wherein the ratio of a pharmaceutically acceptable salt and/or co-crystal of sepiapterin to the antioxidant is greater than 4:1 (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) by weight (e.g. weight of salt to antioxidant) . The pharmaceutical composition may contain 20-30% of a pharmaceutically acceptable salt and/or co-crystal of sepiapterin based on total weight, for example 20%, 22%, 25%, 27% or 30%. The pharmaceutical composition may further contain a dispersant, such as croscarmellose sodium. The pharmaceutical composition may contain 0.1-1.5% dispersant relative to the total weight, for example 0.1, 0.5, 1 or 1.5%. In some embodiments, the pharmaceutical formulation contains at least one anti-caking agent, such as colloidal silicon dioxide or microcrystalline cellulose. The pharmaceutical composition may contain 6575% anti-caking agent relative to the total weight, for example 65%, 67%, 70%, 73% or 75%. In some embodiments, the pharmaceutical composition contains both colloidal silicon dioxide and microcrystalline cellulose. In some embodiments, the pharmaceutical composition contains 60-65% microcrystalline cellulose based on total weight and 5-7% collo

- 23 042976 id silicon dioxide relative to the total weight. In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin is formulated as a particle size less than 140 microns, such as 120, 110, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, or 5 µm. In some embodiments, the pharmaceutical composition contains less than 1.3%, for example, less than 1%, of an impurity such as lactoylpterin, for example, 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%.

A pharmaceutically acceptable salt and/or co-crystal of sepiapterin may serve as a useful therapeutic agent for diseases associated with low intracellular BH4 levels or dysfunction of various BH4 dependent metabolic pathways, including, but not limited to, primary tetrahydrobiopterin deficiency, GTPCH deficiency, 6-pyruvoyltetrahydropterin synthase (PTPS) deficiency ), DHPR deficiency, sepiapterin reductase deficiency, dopamine-dependent dystonia, Segawa syndrome, tyrosine hydroxylase deficiency, phenylketonuria, DNAJC12 deficiency, Parkinson's disease, depression caused by Parkinson's disease, impulsivity in patients with parkinsonism, major depression, autism spectrum, ADHD, schizophrenia, bipolar disorder, cerebral ischemia, restless legs syndrome, obsessive-compulsive disorder, anxiety, aggression in Alzheimer's disease, cerebrovascular disorders, spasm after subarachnoid hemorrhage, myocarditis, coronary vasospasm, cardiac hypertrophy, arteriosclerosis, hypertension, thrombosis, infections, endotoxin shock, liver cirrhosis, hypertrophic pyloromonofunctional stenosis, damage to the gastric mucosa, pulmonary hypertension, impaired renal function, impotence and hypoglycemia. Thus, various forms of sepiapterin in accordance with the present invention can be administered to a patient in an effective amount to provide treatment or alleviation of a disease, disorder or condition.

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

In some embodiments, the pharmaceutical composition contains a crystalline salt and/or co-crystal of sepiapterin. The crystalline salt and/or co-crystal of sepiapterin may be present as an anhydride (eg without any bound water or solvent or hydration or solvation) or as a hydrate, partial hydrate (eg hemihydrate, sesquihydrate), dihydrate, trihydrate, where the crystalline form binds water of hydration or a solvent molecule associated with the crystalline form of sepiapterin or its salt and/or co-crystal. In one embodiment, the crystalline salt and/or co-crystal of sepiapterin is present as a monohydrate or as a hemihydrate.

The present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount, for example a therapeutically effective amount, including a prophylactically effective amount, of a pharmaceutically acceptable salt and/or co-crystal of sepiapterin.

The pharmaceutically acceptable carrier may be any of the commonly used carriers and is limited only by chemical-physical considerations such as solubility and non-reactivity with the compound, and the route of administration. A person skilled in the art will appreciate that, in addition to the following pharmaceutical compositions described, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin may be formulated as inclusion complexes, such as cyclodextrin inclusion complexes, or liposomes.

The pharmaceutically acceptable carriers described in the present invention, such as carriers, adjuvants, excipients or diluents, are well known to those skilled in the art and readily available to the public. Preferably, the pharmaceutically acceptable carrier is chemically inert with respect to the active compounds and the carrier does not have harmful side effects or toxicity under the conditions of use.

Dosage.

The pharmaceutically acceptable salt of sepiapterin may be used at any suitable dosage. Suitable doses and dosing regimens can be determined by conventional ranging methods. Typically, treatment is started with smaller doses that are less than the optimal dose. Thereafter, the dosage is increased in small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dose may be divided and, if desired, administered in portions throughout the day. At the right doses, and with the appropriate administration of certain compounds, the present invention provides a wide range of responses. Typically, a dosage range of about 2.5 to about 150 mg/kg of the patient's body weight is administered per day. For example, in embodiments, a pharmaceutically acceptable salt of sepiapterin may 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, from about 80 mg/kg to about 100 mg/kg, from about 40 mg/kg to about 60 mg/kg, from about 2.5 mg/kg to about 20 mg/kg, from about 2, 5 mg/kg to about 10 mg/kg, from about 2.5 mg/kg to about 5 mg/kg, of the subject's body weight per day, one or more times per day, to obtain the desired therapeutic effect.

In some embodiments, the dose is an amount sufficient to generate CNS levels of BH4, such as as measured in cerebrospinal fluid and/or brain and/or sufficient to produce therapeutic results, such as an increase in CNS serotonin or dopamine levels. In some embodiments, the dose is an amount sufficient to increase BH4 levels to at least twice the pre-administration BH4 levels, as measured in the subject's plasma or organ, e.g., the subject's liver.

In some embodiments, the pharmaceutically acceptable salt and/or co-crystal of sepiapterin may be formulated into a solid dosage form such as particles. In these embodiments, each unit solid dosage form may contain any suitable amount of a pharmaceutically acceptable salt and/or co-crystal of sepiapterin. For example, each solid dosage form may 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.

Ways of introduction.

The choice of carrier will be determined in part by the particular active agent as well as the particular route used to administer the composition. Accordingly, a wide variety of suitable formulations of the pharmaceutical composition of the present invention are provided. The following formulations for oral, aerosol, parenteral, subcutaneous, intravenous, intra-arterial, intramuscular, intraperitoneal, intrathecal, rectal, and vaginal administration are merely illustrative and are in no way limiting.

The pharmaceutical composition may be a liquid formulation such as a solution, suspension or emulsion. Formulations suitable for oral administration may consist of (a) capsules, sachets, tablets, lozenges and troches, each containing a predetermined amount of the active ingredient in the form of solid particles or granules; (b) powders; (c) liquid solutions, such as an effective amount of the compound dissolved in diluents such as water, saline, or orange juice; (e) suspensions in an appropriate liquid; and (e) suitable emulsions. Solid oral dosage forms such as capsules, tablets and powders are preferred. Capsules may be of the conventional type, hard or soft shell gelatin containing, for example, surfactants, lubricants, and excipients such as lactose, sucrose, calcium phosphate, and corn starch. Tablets may contain one or more of: lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, gum, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate , stearic acid and other excipients, coloring agents, diluents, buffering agents, disintegrating agents, wetting agents, preservatives, flavoring agents and pharmacologically compatible carriers. Lozenges may contain the active ingredient in a flavoring agent, usually sucrose and gum arabic or tragacanth, and lozenges may contain the active ingredient in an inert base such as gelatin and glycerin, or sucrose and gum arabic, emulsions, gels and the like, containing, in addition to the active ingredient , media that are known in the art.

Formulations suitable for oral and/or parenteral administration include aqueous and non-aqueous, isotonic sterile injectable solutions, which may contain antioxidants, buffers, bacteriostatics, and solutes that render the formulation isotonic with respect to the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions, which may contain suspending agents, solubilizers, thickeners, stabilizers and preservatives. The compound may be administered in a physiologically acceptable diluent in a pharmaceutical carrier such as a sterile liquid or mixture of liquids including water, saline, aqueous dextrose and related sugar solutions, alcohol such as ethanol, benzyl alcohol or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol and other polyethylene alcohols, ketals of glycerol such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers such as poly(ethylene glycol) 400, oil, fatty acid, ester or fatty acid glyceride or an acetylated fatty acid glyceride with or without the addition of a pharmaceutically acceptable surfactant such as soap or detergent, a suspending agent such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose or carboxymethylcellulose, or emulsifiers and other pharmaceutical adjuvants.

- 25 042976

Oils that can be used in parenteral formulations include petroleum, animal, vegetable or synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, vaseline, and mineral oils. Suitable fatty acids 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 alkali metal, ammonium and triethanolamine fatty salts, and suitable detergents include (a) cationic detergents such as, for example, dimethyldialkyl ammonium halides and alkyl pyridinium halides, (b) anionic detergents, such as, for example, alkyl , aryl, olefinsulfonates, alkyl, olefin, ether and monoglyceride sulfates, and sulfosuccinates, (c) non-ionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides and polyoxyethylene-polypropylene copolymers, (e) amphoteric detergents, such as, for example , alkyl-beta-aminopropionates, and 2-alkyl-imidazolium quaternary ammonium salts, and (3) mixtures thereof.

Parenteral formulations will typically contain from about 20% to about 30% by weight of sepiapterin or a pharmaceutically acceptable salt and/or co-crystal thereof in solution. Suitable preservatives and buffers may be used in these formulations. To minimize or eliminate irritation at the injection site, such compositions may contain one or more non-ionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in a given formulation varies from about 5 to approximately 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters such as sorbitan monooleate, and high molecular weight adducts of ethylene oxide with a hydrophobic base formed by the condensation of propylene oxide with propylene glycol. Parenteral formulations may be presented in single-dose or multi-dose sealed containers such as ampoules and vials and may be stored under lyophilized conditions requiring only the addition of a sterile liquid carrier, eg water, for injection immediately prior to use. Solutions for injections and suspensions for immediate administration can be prepared from sterile powders, granules and tablets of the types previously described.

The pharmaceutical composition may be an injectable formulation. Requirements for 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. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA.

Topical formulations, including formulations that are suitable for transdermal drug release, are well known to those skilled in the art and are suitable in the context of the present invention for application to the skin. Topically applied compositions are usually used in the form of liquids, creams, pastes, lotions and gels. Topical administration includes application to the oral mucosa, which includes the oral cavity, oral epithelium, palate, gums, and nasal mucosa. In some embodiments, the composition comprises sepiapterin and a suitable medium or carrier. It may also contain other ingredients such as an anti-inflammatory agent. The carrier may be liquid, solid or semi-solid. In embodiments, the composition is an aqueous solution. Alternatively, the composition may be a dispersion, emulsion, gel, lotion or cream carrier for the various components. In one embodiment, the base medium is water or a biocompatible solvent that is essentially neutral, or that has been found to be essentially neutral. The liquid medium may contain other materials such as buffers, alcohols, glycerin, and mineral oils with various emulsifiers or dispersing agents, as is known in the art, to obtain the desired pH, consistency, and viscosity. It is possible that the compositions may be prepared as solids such as powders or granules. Solids can be applied directly or dissolved in water or a biocompatible solvent prior to use to provide a solution that is essentially neutral or that is recognized as being essentially neutral and which can then be applied to the target site. In embodiments of the present invention, the topical skin application medium may include water, buffer solutions, various alcohols, glycols such as glycerol, lipid materials such as fatty acids, mineral oils, phosphoglycerides, collagen, gelatin, and silicone-based materials.

The pharmaceutical composition may be an aerosol formulation for administration by inhalation. These aerosol formulations can be placed in pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen, and the like. They can also be formulated as non-pressure pharmaceuticals such as an inhaler or nebulizer.

In addition, the pharmaceutical composition may be a suppository. Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or sprays containing, in addition to the active ingredient, such carriers as are known in the art to be suitable.

- 26 042976

Solid dosage form for oral administration.

Formulations for oral use contain particles containing the active ingredient(s) in admixture with non-toxic pharmaceutically acceptable excipients and these formulations are known to those skilled in the art (e.g., US Pat. 6036949, 6036949, 6372218, included in the present invention by reference). Excipients may be, for example, inert diluents or fillers (eg, sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starches, including corn starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulating agents and disintegrants (eg, cellulose derivatives, including microcrystalline cellulose, starches, including corn starch, croscarmellose sodium, alginates, or alginic acid); binders (eg sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricating agents, glidants, release agents (eg, magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils, or talc), and anti-caking agents (eg, colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, stearate magnesium, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, powdered talc, sodium aluminum silicate, potassium aluminum silicate, calcium aluminum silicate, bentonite, aluminum silicate, stearic acid, polydimethylsiloxane ). Other pharmaceutically acceptable excipients may be colorants, flavoring agents, plasticizers, humectants and buffering agents. In some embodiments, excipients (eg, flavoring agents) are packaged with the composition. In some embodiments, excipients (eg, flavors) are packaged separately from the composition (eg, combined with the composition prior to administration).

The solid compositions of the present invention may include a coating adapted to protect the composition from undesirable chemical changes (eg, chemical degradation prior to release of the active substances). The coating can be applied to the solid dosage form in the same manner as described in Remington: The Science and Practice of Pharmacy, (22nd ed.) ed. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA.

Powders and granulates can be prepared by using the ingredients mentioned above in a conventional manner using, for example, a mixer, fluid bed apparatus, melt solidifier, rotary granulator, extruder/spheronizer or spray drying equipment.

Methods of treatment.

The present invention relates to pharmaceutical compositions, for example, in an orally tolerated form, which contains a therapeutically effective amount of a pharmaceutically acceptable salt and/or co-crystal of sepiapterin, for example, and less than 10% of an antioxidant. In some embodiments, the pharmaceutical composition is a granular composition that is dispersed in a pharmaceutically acceptable carrier, eg, the composition can be mixed in water and swallowed by a patient (eg, within 5-10 minutes). Suitable formulations for use in the present invention can be found in Remington: The Science and Practice of Pharmacy, (22nd ed.) ed. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA. Except where any conventional carrier is incompatible with the active ingredient, its use in pharmaceutical compositions is contemplated. Moreover, for administration to animals (eg, humans), it should be understood that formulations must meet standards of sterility, pyrogenicity, general safety, and purity as required by regulatory authorities.

The actual dose of the composition 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 idiopathy, and route of administration. Depending on the dosage and route of administration, the number of administrations of the preferred dosage and/or effective amount may vary depending on the response of the subject. The physician responsible for administration will in any event determine the concentration of the active ingredient(s) in the composition and the appropriate dose(s) for the individual subject.

In some embodiments, patients receive 2.5, 5, 10, 20, 40, 60, or 80 mg/kg/day of a pharmaceutically acceptable salt and/or sepiapterin co-crystal. Patients may receive a pharmaceutical composition containing sepiapterin once a day, twice a day, or three times a day during treatment. In some embodiments, patients continue to take other current drugs for a BH4 disorder (e.g., L-dopa/carbidopa, 5HTP, melatonin, MAO inhibitors, and dopamine receptor agonists as prescribed), except for BH4 supplements (if they take BH4). Patients may be prohibited

- 27 042976 take any drugs that inhibit the synthesis of folic acid (for example, methotrexate, pemetrexed or trimetrexate).

In some embodiments, patients receiving BH4 therapy prior to study entry undergo a washout period during screening prior to administration of the pharmaceutical composition of the present invention. Patients may be advised to follow a consistent diet for protein and phenylalanine (Phe) intake. Diet records may be reviewed by a qualified dietitian. Total Phe concentrations over a 3-day period can be calculated by a nutritionist and recorded.

In some embodiments, patients taking BH4 stop taking BH4 (i.e. washout of BH4). Blood samples for determination of Phe concentrations may be obtained during the BH4 washout period 7, 5, 3 and 1 days prior to treatment with the pharmaceutical composition of the present invention or until blood Phe levels are >360 µmol/L at any time in washout period of BH4. In some embodiments, a blood sample is tested for sepiapterin, Phe, BH4, and tyrosine (Tyr) prior to dosing.

Methods for obtaining compositions.

In some embodiments, the pharmaceutical composition of the present invention can be prepared by mixing sepiapterin or a pharmaceutically acceptable salt and/or co-crystal thereof and an antioxidant with one or more excipients, such as a dispersant, and one or more anti-caking agents. In some embodiments, each of the components of the composition is passed through a size exclusion filter (eg, a filter having pores of 200 microns or less) prior to mixing. In some embodiments, the anti-caking agents are mixed together before the components (eg, sepiapterin, dispersant, and antioxidant) are added.

In some embodiments, the pharmaceutical composition is obtained:

(a) passing at least one anti-caking agent through a size exclusion filter (eg, a filter with a pore size of less than 200 microns);

(b) mixing the sepiapterin, the antioxidant, and optionally the dispersant with at least one anti-caking agent (eg, by mixing in a blender); and (c) passing the combination of step b through a size exclusion filter (eg, a filter with a pore size of less than 150 microns).

In some embodiments, the at least one step a anti-caking agent includes more than one anti-caking agent (eg, two anti-caking agents) that are mixed together before being passed through the size exclusion filter.

Examples

Although some features of the present invention have been illustrated and described in the present invention, many modifications, substitutions, changes and equivalents will be apparent to those skilled in the art. Therefore, it is to be understood that the appended claims are intended to cover all such modifications and changes that fall within the true spirit of the present invention. As such, the following examples are presented to disclose various aspects of the present invention. These examples represent individual embodiments of aspects of the present invention, and one skilled in the art will appreciate that additional examples can be obtained to equally disclose aspects of the present invention.

Example 1 Preparation of sepiapterin salts.

Salts and/or co-crystals of sepiapterin and 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 suspending sepiapterin as free bases and acids in acetone/water (9/1, v/v) or methanol for 2-17 days.

Salts were analyzed by DSC, TGA, HPLC, IR and RPD. The results are shown in table. 15 below. The IR spectra are shown in Fig. 1-13.

Table 15. Summary of analyzes of the salt and/or co-crystal of sepiapterin

Salt form weight (mg) TGA a loss weight (%) dec endothera (°C, start) purity (%) Molar ratio (FB: acid) Residual solvent HC1 salt 190.9 3.6 218.3 93.94 1:1.3 Minor amount Acetone Methansul 188.0 4.23 182.3 91.14 1:1.0 negligible

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phonet th quantity Meon Nicotinate 246.0 1.27 220.4 97.16 1:0.9 No Toluolsul fonate 256.4 0.5 190.3 262.9 96.84 1:1.0 No Benzenesulfonate 173.3 1.54 192.7, 206.2 90.20 1:1.0 Minor quantity Meon Sulfate 227.5 2.6 196.5 97.33 1:0.6 No Phosphate 235.8 AND 2 144.0, 206.8 96.75 1:1.1 No Malonat 95.8 3.83 175.1 99.45 needs clarification No L-Tartrate 232.2 1.14 156.5, 174.6 99.75 1:1.0 Minor amount Acetone Fumarate 217.3 4.81 77.3, 132.8, 190.1 99.46 1:0.6 Minor amount Acetone Gentizat 98.1 6.92 83.2 133.8, 149.0 94.35 1:0.5 No glycolate 135.4 20.15 79.3, 90.0, 132.3, 151.6 99.19 1:0.3 No

PB=free base.

Example 2 Stability analysis of salts and/or co-crystals of sepiapterin.

The stability of the obtained salts and/or co-crystals was analyzed after 1 and 2 weeks at 25°C and 60% relative humidity and 40°C and 75% relative humidity. The results are shown in table. 16 below. Surprisingly, of all the salts and/or co-crystals tested, the phosphate salt and/or co-crystal, tartrate salt and/or co-crystal, and nicotinate salt and/or co-crystal were markedly more stable than the others. None of the phosphate, tartrate, nicotinate salt and/or co-crystals underwent a shape change during stability testing, and each maintained greater than 97% purity during the two weeks of the study. In fact, tartrate and nicotinate retained over 99% purity.

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Table 16. Summary of Stability Study Results

Salt Moment time conditions Shape change purity (square%) Purity relative to original. (%) Phosphate Initially NA ΝΑ 96.75 ΝΑ 1 Week 25°C/60%RH No 95.35 98.6 40°C/75%RH No 95.91 99.1 2 weeks 25°C/60%RH No 95.87 99.1 40°C/75%RH No 94.50 97.7 L-Tartrate Initially ΝΑ ΝΑ 99.75 ΝΑ 1 Week 25°C/60%RH No 98.61 99.9 40°C/75%RH No 99.06 99.3 2 weeks 25°C/60%RH No 99.39 99.6 40°C/75%RH No 99.00 99.3 glycolate Initially NA ΝΑ 99.19 ΝΑ 1 Week 25°C/60%RH Glycolate and free base 98.93 99.7 40°C/75%RH Glycolate and free base 98.54 99.3 2 weeks 25°C/60%RH Glycolate and free base 98.86 99.7

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Salt Moment time conditions Change forms purity (square%) Purity relative to original. (%) 40°C/75%RH Glycolate and free base 98.52 99.3 Fumarate Initially ΝΑ NA 99.46 NA 1 Week 25°C/60%RH No 99.39 99.9 40°C/75%RH No 99.15 99.7 2 weeks 25°C/60%RH Fumarate and free base 99.25 99.7 40°C/75%RH Fumarate and free base 98.98 99.5 Gentizat Initially NA NA 94.35 NA 1 Week 25°C/60%RH Gentisate and free base 97.66 103.5 40°C/75%RH Gentisate and free base 96.89 102.7 2 weeks 25°C/60%RH Gentisate and free base 97.00 102.8 40°C/75%RH Gentisate and free base 93.37 102.1 Malonat Initially NA NA 99.45 NA

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Salt Moment time conditions Change forms purity (square%) Purity relative to original.(%) 1 Week 25°C/60%RH Malonate and free base 99.39 99.9 40°C/75%RH Malonate and free base 99.14 99.7 2 weeks 25°C/60%RH Malonate and free base 99.23 99.8 40°C/75%RH Malonate and free base 97.81 98.3 HC1 Initially ΝΑ NA 93.94 NA 1 Week 25°C/60%RH No 97.19 103.5 40°C/75%RH No 89.25 95.0 2 weeks 25°C/60%RH No 91.84 97.8 40°C/75%RH No 84.16 89.6 Methansul fonat Initially NA NA 91.14 NA 1 Week 25°C/60%RH No 95.26 104.5 40°C/75%RH No 88.68 97.3 2 weeks 25°C/60%RH No 91.95 100.9 40°C/75%RH No 85.97 94.3 Nicotine t Initially NA NA 97.16 NA 1 Week 25°C/60%RH No 97.43 100.3 40°C/75%RH No 97.30 100.2 2 weeks 25°C/60%RH No 97.45 100.3

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Salt Moment time conditions Shape change purity (square%) Purity relative to original.(%) 40°C/75%RH No 97.41 100.3 Toluene fonate Initially ΝΑ ΝΑ 96.84 ΝΑ 1 Week 25°C/60%RH No 94.19 97.3 40°C/75%RH No 89.11 92.0 2 weeks 25°C/60%RH No 91.40 94.4 40°C/75%RH No 88.12 91.0 Benzene su lfonate Initially NA ΝΑ 90.20 ΝΑ 1 Week 25°C/60%RH No 90.68 100.5 40°C/75%RH No 82.63 91.6 2 weeks 25°C/60%RH No 86.37 95.8 40°C/75%RH No 82.65 91.6 Sulfate Initially NA ΝΑ 97.33 ΝΑ 1 Week 25°C/60%RH No 95.22 97.8 40°C/75%RH No 89.44 91.9 2 weeks 25°C/60%RH No 93.46 96.0 40°C/75%RH No 88.20 90.6

NA=Not provided, RH=relative humidity.

Example 3 Solubility and disproportionation of various salts and/or co-crystals of sepiapterin.

Kinetic solubility was evaluated for the nicotinate, phosphate, L-tartrate and fumarate salts and/or sepiapterin co-crystals in water and Medisca oral formula. Powder X-ray diffraction (XRD) was performed on the residual solids to detect shape change/disproportionation. The solids were suspended in medium at a target concentration of ~7 mg/mL (calculated as free base). The suspensions were stirred in a rotating incubator at 25 rpm for 1, 4 and 24 hours. At each time point, 1 ml of the suspension was pipetted for centrifugation at 10,000 rpm (2 min) and filtration through a 0.45 μm membrane to obtain a supernatant for HPLC and pH dissolution tests, residual solids were analyzed by RPD. The solubility results are presented in table. 17-20.

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Table 17 Solubility Summary of Nicotinate Salt and/or Co-Crystal

Wednesday Moment of time u(h) Pace •(°C) Solubility (mg/ml)* Purity (area%) ^# Observation Change forms pH water 1 RT 1.5 98.97 Muti th Yes 2.1 4 2.3 99.04 Muti th Yes 2.1 24 1.8 96.54 Muti th Yes 2.1 Medisca Oral Mixture 1 2.6 99.76 Muti th Yes 3.1 4 3.1 99.60 Muti th Yes 3.1 24 3.5 97.00 Muti th Yes 3.1

*calculated using free base, ^# excess salt sample was dosed for solubility measurement, which may result in enrichment of impurities in the supernatant, so purity data is for reference only.

Table 18. Solubility summary of phosphate salt and/or co-crystal

Wednesday Moment of time u(h) Temp (°C) Solubility (mg/ml)* Purity (area %) ^# observation Shape change pH water 1 RT 1.7 89.92 Muddy Yes 2D 4 2.0 89.52 Muddy Yes 2.1 24 1.9 82.64 Muddy Yes 2.1 Mixture 1 2.5 99.23 Mutny Yes** 3.1 oral Medisca th 4 3.2 98.95 Muddy Yes** 3.1 24 2.1 87.63 Muddy Yes** 3.1

*calculated using free base, ** low crystallinity, ^# excess salt sample was dosed for solubility measurement, which may result in enrichment of impurities in the supernatant, so purity data is for guidance only.

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Table 19 Solubility Summary of L-Tartrate Salt and/or Co-Crystal

Wednesday Point in time (h) Pace. (°C) Solubility (mg/ml) * Purity (area %) ^# observed not Shape change pH water 1 RT 1.6 99.34 Turbid Yes 2.5 4 1.8 99.07 Turbid Yes 2.5 24 1.8 95.61 Turbid Yes 2.5 Medisca Oral Mixture 1 2.0 99.68 Turbid Yes 3.3 4 2.5 99.54 Turbid Yes 3.3 24 3.2 95.67 Turbid Yes 3.3

*calculated using the free base, #

an excess amount of the salt sample was dosed to measure solubility, which may lead to enrichment of impurities in the supernatant, so the purity data is for reference only.

Table 20. Solubility summary of fumarate salt and/or co-crystal

Wednesday Moment of time (h) Pace. (°C) Solubility (mg/ml)* Purity (area %)* Observation Shape change pH water 1 RT 1.2 98.39 Turbid No 3.1 4 1.4 98.19 Turbid No 3.1 24 1.5 95.43 Turbid No 3.1 Medisca Oral Mixture 1 2.2 99.74 Turbid No** 4.0 4 3.1 99.60 Turbid No** 4.0 24 2.9 96.54 Turbid No** 4.0

*calculated using free base, ** low crystallinity, # excess salt sample was dosed for solubility measurement, which may result in enrichment of impurities in the supernatant, so purity data is for guidance only.

Results: For nicotinate, phosphate and L-tartrate salts and/or co-crystals, residual solids converted to free base in water and Medisca oral formula after 1 hour. water, while the crystallinity of the residual solids decreased after 1 hour in the Medisca oral mixture. Surprisingly, of the twelve different salt forms and/or co-crystals studied, the fumarate salt and/or co-crystal was the only salt and/or co-crystal found to be highly stable in the solid form stability study of Example 2, and no evidence of disproportionation was found in research on disproportionation.

Other embodiments

Although the present invention has been described in connection with its specific embodiments, it should be understood that it is open to further modifications, and this application is intended to cover any variations, uses or adaptations of the present invention, following generally the principles of the present invention and including such deviations from the present invention. inventions that are known or common practice in the technical field to which the present invention relates, and can be applied to the essential features set forth in the present invention above.

Claims (10)

1. A pharmaceutically acceptable salt of sepiapterin in amorphous or crystalline form, wherein the pharmaceutically acceptable salt is a nicotinate salt or a tartrate salt. 2. The pharmaceutically acceptable salt of claim 1, wherein the pharmaceutically acceptable salt is the nicotinate salt. 3. The pharmaceutically acceptable salt of claim 1, wherein the pharmaceutically acceptable salt is the tartrate salt. 4. A pharmaceutically acceptable salt according to any one of claims 1 to 3, wherein the salt is crystalline. 5. Pharmaceutical composition containing a pharmaceutically acceptable salt according to any one of paragraphs. 1-4 and a pharmaceutically acceptable excipient. 6. A method of treating a disorder associated with tetrahydrobiopterin in a subject in need thereof; and/or increasing levels of tetrahydrobiopterin in a subject in need thereof; and/or reducing levels of phenylalanine in a subject in need thereof; and/or increasing phenylalanine hydroxylase activity in a subject; and/or treatment of phenylketonuria in a subject in need thereof; and/or increasing serotonin levels in a subject in need thereof; and/or increasing tryptophan hydroxylase activity in a subject; and/or increasing dopamine levels in a subject in need thereof; and/or increasing tyrosine hydroxylase activity in a subject; and/or increasing nitric oxide synthase activity in a subject; and/or increasing alkylglycerol monooxygenase activity in a subject; and/or increasing the level of homovanic acid and/or 5-hydroxyindoleacetic acid in the subject, wherein the method comprises administering to the subject an effective amount of a pharmaceutically acceptable salt according to any one of claims 1 to 4 or a pharmaceutical composition according to claim 5. 7. The method of claim 6 wherein the tetrahydrobiopterin related disorder is phenylketonuria or tetrahydrobiopterin deficiency. 8. The method according to claim 6 or 7, wherein an effective amount of a pharmaceutically acceptable salt according to any one of claims 1 to 4 or a pharmaceutical composition according to claim 5 comprises an amount sufficient to increase the level of tetrahydrobiopterin in plasma, cerebrospinal fluid (CSF) and/or brain of the subject at least twice as compared with the level of tetrahydrobiopterin before administration. 9. The method of claim 8, wherein the level of homovanilic acid and/or 5-hydroxyindoleacetic acid in the CSF of the subject is increased. 10. The method according to any one of claims 6-9, wherein the subject is a human.