Compositions and methods for increasing tetrahydrobiopterin plasma exposure

Abstract

To provide compositions including sepiapterin, or a pharmaceutically acceptable salt thereof, and methods for the treatment of BH4-related disorders.SOLUTION: The present invention provides a method of treating a BH4-related disorder in a subject in need thereof, the method comprising the step for administering to the subject an effective amount of sepiapterin or a pharmaceutically acceptable salt thereof, together with food. In some embodiments, these compositions and methods result in an increase in plasma exposure of BH4.SELECTED DRAWING: None

Description

[Background technology]

[0001] Sepiapterin is a naturally occurring precursor of tetrahydrobiopterin (BH4), a naturally occurring essential cofactor for important intracellular enzymes (including, but not limited to, phenylalanine hydroxylase (PAH) (Kaufman, 1958), tyrosine hydroxylase (TH) (Nagatsu et al, 1964), tryptophan hydroxylase (TPH) (Ichiyama et al, 1970), nitrile oxide synthase (NOS) (Kwon et al, 1989), (Mayer et al, 1991), and alkylglycerol monooxygenase (AGMO) (Tietz et al, 1964). The rapid conversion of sepiapterin to BH4 occurs via a two-step reduction in the salvage pathway for BH4 synthesis (Sawabe, 2008). Synthetic forms of BH4 (e.g., sapropterin dihydrochloride) are used as a therapy for hyperplasmic phenylalanine-related disorders such as phenylketonuria (PKU). PKU is a congenital metabolic disorder primarily caused by mutations in the PAH gene. BH4 has also been tested as a treatment for various central nervous system symptoms and other disorders associated with PKU, but its efficacy has been limited, possibly because BH4 cannot effectively cross the blood-brain barrier (Klaiman et al, 2013, Grant et al, 2015).

[0002] Recent studies suggest that, compared to BH4, peripherally administered sepiapterin has higher membrane permeability, resulting in easier access to liver, kidney, and brain cells. Sepiapterin has been reported to be rapidly converted to BH4 intracellularly, thereby increasing BH4 levels in the liver, kidney, and brain (Sawabe, 2008). As a result, sepiapterin may serve as a useful therapeutic agent for diseases associated with low intracellular BH4 levels or dysfunction of various BH4-dependent metabolic pathways. [Overview of the project]

[0003] This invention aims to discover that when sepiapterin is administered to subjects with food, BH4 production unexpectedly increases, and further, the subjects become exposed to BH4 plasma, CSF, and / or brain. While not bound by theory, increased BH4 plasma exposure may lead to delayed absorption of sepiapterin due to increased gastric residence time, or prolonged absorption rate of sepiapterin due to increased foregut residence time, or a maximal rate of enzyme activity (V) of sepiapterin reductase or dihydrofolate reductase. max Extended intracellular concentrations of sepiapterin below, equal to, or slightly above ) may result from a combination of the above, leading to a greater proportion of administered sepiapterin being converted to BH4 before passive or active transport into the circulation for elimination and / or removal. The present invention features compositions comprising sepiapterin or salts thereof, and methods for treating BH4-related diseases. In some embodiments, the compositions and methods result in increased plasma exposure to BH4.

[0004] As is known in the art, food can affect the absorption of compounds. Absorption may be delayed but not reduced, or the total amount of drug absorbed may be reduced. The effect of food may be due to slowing gastric retention time, slowing foregut retention time, reducing access of the compound to the absorption site, altering the dissolution rate of the compound, or altering the pH of the stomach. Because of this effect, it is important to establish a specific administration schedule for drugs that should be administered separately from or with food.

[0005] In one embodiment, the present invention is characterized by a method for treating BH4-related diseases in subjects requiring such treatment, by administering an effective amount of sepiapteprine or a pharmaceutically acceptable salt thereof together with food.

[0006] In another embodiment, the present invention is characterized by a method for increasing BH4 plasma exposure in a subject receiving sepiapterin therapy by administering an effective amount of sepiapterin or a pharmaceutically acceptable salt thereof to the subject together with food.

[0007] In a further embodiment, the present invention is characterized by a method for reducing the absorption rate of an oral formulation of sepiapterin, as measured by the BH4 concentration that reaches the plasma over time in a subject requiring its therapeutic effect. The method comprises administering an effective amount of sepiapterin or a pharmaceutically acceptable salt thereof to the subject together with food.

[0008] In some embodiments of any of the methods described above, the effective dose is a dose sufficient to produce a BH4 concentration of at least 50 ng / mL (e.g., at least 60 ng / mL, at least 100 ng / mL, at least 200 ng / mL, at least 400 ng / mL, at least 600 ng / mL, at least 1000 ng / mL, or at least 2000 ng / mL) in the target plasma within 10 hours after administration with food (e.g., 2.5 mg / kg to 100 mg / kg per administration). The effective dose may include a dose that is at least 5% (at least 10%, at least 20%, at least 50%, at least 70%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%) less than the dose sufficient to produce a maximum BH4 plasma concentration (Cmax) of at least 50 ng / ml (e.g., at least 60 ng / mL, at least 100 ng / mL, at least 200 ng / mL, at least 2000 ng / mL) in the subject's plasma within 10 hours after administration of sepiapterin without food.

[0009] In some embodiments of any of the methods described above, administration to the subject is performed less than 30 minutes before food intake or after food intake, for example, from immediately before food intake to 1 hour after intake. In some embodiments, administration to the subject is performed substantially simultaneously with food. In some embodiments of any of the methods described above, the food is a high-protein food. In some embodiments of any of the methods described above, the food is a high-fat food (for example, at least 25, 30, 40, or 50% of the calories come from fat). In some embodiments of any of the methods described above, the food is a high-protein and high-fat food. In some embodiments, the food is a high-calorie food (for example, the food contains at least 100 calories, for example, at least 200 calories, at least 300 calories, at least 400 calories, at least 500 calories, for example, 500-1500 or 800-1000 calories). In some embodiments of any of the methods described above, the food is a meal, for example, breakfast, lunch, or dinner.

[0010] In some embodiments, administration with food (e.g., less than 30 minutes before food intake or after food intake, e.g., immediately before food intake to 1 hour after intake) results in an increase in BH4 Cmax (e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%) compared to administration without food (e.g., more than 2 hours after food intake but up to 30 minutes before further food intake).

[0011] In some embodiments, administration with food (e.g., less than 30 minutes before food intake or after food intake, e.g., immediately before food intake to 1 hour after intake) is more effective than administration without food (e.g., less than 30 minutes before food intake or after food intake, e.g., immediately before food intake to 1 hour after intake) in terms of the degree of BH4 production and the resulting plasma exposure (AUC). 0-lastThis results in an increase (for example, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%).

[0012] In some embodiments of any of the above embodiments, the sepiapterin is provided in a composition separate from the food to be ingested (for example, the sepiapterin is not incorporated into the food product). In some embodiments of any of the above embodiments, the food is ingested before the administration of the sepiapterin (for example, the food is ingested within one hour immediately before the administration of the sepiapterin). In some embodiments, the food is ingested after the administration of the sepiapterin (for example, the food is ingested between immediately after administration and 30 minutes after administration).

[0013] In a further embodiment, the present invention features pharmaceutical compositions of sepiapterin or a pharmaceutically acceptable salt thereof, which are formulated to mimic the effects of administration with food, for example, to increase gastric retention time (e.g., formulations described in Radhakrishnan et al. Drug Delivery Letters, 2017, 7, 190-200, which are incorporated by reference).

[0014] In some embodiments, the composition is formulated as a bioadhesive formulation, a high-density formulation, a swelling formulation, a superporous hydrogel formulation, or a floating formulation (for example, a composition comprising an ion exchange resin, a raft system, an inflatable chamber, a foaming mixture, a swelling hydrocolloid, or a multi-particle system).

[0015] In another embodiment, the present invention provides a method for treating BH4-related diseases in subjects requiring such treatment, characterized by administering an effective amount of any of the above-mentioned pharmaceutical compositions to the subject.

[0016] In yet another embodiment, the present invention is characterized by a method of increasing BH4 plasma exposure in a subject receiving sepiapterin therapy by administering an effective amount of any of the above-described pharmaceutical compositions to the subject.

[0017] In a further embodiment, the present invention is characterized by a method of administering an effective amount of any of the above-mentioned pharmaceutical compositions to a subject to delay or reduce (for example, reduce by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%) the absorption rate of an oral formulation of sepiapterin, as measured by the BH4 concentration reached in the plasma over time in a subject requiring its therapeutic effect.

[0018] In one embodiment, the present invention is characterized by a method for increasing the levels of homovanillic acid and / or 5-hydroxyindoleacetic acid in a subject, the method comprising the step of administering an effective amount of sepiapterin or a pharmaceutically acceptable salt thereof together with a food. In some embodiments, the levels of homovanillic acid and / or 5-hydroxyindoleacetic acid in the cerebrospinal fluid (CSF) of the subject are increased. In some embodiments, the levels of homovanillic acid and / or 5-hydroxyindoleacetic acid in the subject (e.g., in the CSF of the subject) are increased by at least 5% compared to the level before administration (e.g., an increase of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, at least 200%, at least 250%, or at least 300% compared to the level before administration).

[0019] In some embodiments, prior to administration of sepiapterin or a pharmaceutically acceptable salt thereof, the subject has levels of homovanillic acid and / or 5-hydroxyindoleacetic acid that are less than 50% of the average subject level (e.g., less than 40%, less than 30%) (e.g., the subject has a CSF level of homovanillic acid less than 15 ng / ml and / or a CSF level of 5-hydroxyindoleacetic acid less than 5 ng / ml). In some embodiments, the subject has not been diagnosed with BH4-related disease. In some embodiments, the subject does not have symptoms of BH4-related disease. In some embodiments, after administration of sepiapterin or a pharmaceutically acceptable salt thereof, the levels of homovanillic acid and / or 5-hydroxyindoleacetic acid in the subject are greater than 50% of the average subject level (e.g., the subject has a CSF level of homovanillic acid greater than 15 ng / ml and / or a CSF level of 5-hydroxyindoleacetic acid greater than 5 ng / ml).

[0020] In some embodiments of any of the above-described aspects, the subject has and / or has been diagnosed with a BH4-related disease.

[0021] In some embodiments of any of the aspects described above, BH4-related disorders include primary BH4 deficiency, GTPCH deficiency, 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency, DHPR deficiency, sepiapterin reductase deficiency, dopamine-responsive dystonia, Segawa syndrome, tyrosine hydroxylase deficiency, phenylketonuria, DNAJC12 deficiency, Parkinson's disease, depression due to Parkinson's disease, and in patients with Parkinson's disease. These include impulsivity, major depression, autism spectrum disorder, ADHD, schizophrenia, bipolar disorder, cerebral ischemia, restless legs syndrome, obsessive-compulsive disorder, anxiety disorder, aggression in Alzheimer's disease, cerebrovascular disease, convulsions after subarachnoid hemorrhage, myocarditis, coronary spasm, cardiac hypertrophy, arteriosclerosis, hypertension, thrombosis, infection, endotoxin shock, cirrhosis, hypertrophic pyloric stenosis, gastric mucosal injury, pulmonary hypertension, renal dysfunction, impotence, or hypoglycemia. In some embodiments of any of the above-described aspects, BH4-related disorders include phenylketonuria, BH4 deficiency (e.g., primary BH4 deficiency), CNS disorders (e.g., Segawa syndrome, depression, schizophrenia, autism, or Parkinson's disease), or gastrointestinal motility disorders (e.g., gastroparesis and primary and secondary esophageal motility disorders). In some embodiments, BH4-related diseases are BH4 deficiency or phenylketonuria.

[0022] definition In this application, unless otherwise evident from the context, (i) the term “a” may be understood to mean “at least one,” (ii) the term “or” may be understood to mean “and / or,” (iii) the terms “include” and “inclusive” may be understood to include the listed components or processes, whether presented by themselves or together with one or more additional components or processes, and (iv) the terms “about” and “approximately” may be understood to allow for standard variations as understood by those skilled in the art, and (v) if a range is provided, the endpoint is included.

[0023] As used herein, the term "administering" refers to the administration of a composition to a subject. Administration to an animal subject (e.g., a human) may be by any suitable route. For example, in one embodiment, administration is by 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 tracheal instillation), transdermal, vaginal, or intravitreal.

[0024] The "effective amount" of a compound may vary depending on factors such as the individual condition, age, sex, and body weight, as well as the ability of the compound to induce the desired response. A therapeutically effective amount encompasses an amount in which the therapeutically beneficial effects exceed any toxic or detrimental effects of the compound. The effective amount also encompasses an amount sufficient to provide a benefit, such as a clinical benefit.

[0025] As used herein, the term "food" refers to solid food having sufficient bulk and fat content such that it is not rapidly dissolved and absorbed in the stomach. For example, a meal such as breakfast, lunch, or dinner. As used herein, the term "with food" refers to administering the composition between about 30 minutes before eating (e.g., consuming a meal) and about 2 hours after eating. The terms "without food," "fasting," or "on an empty stomach" refer to a state in which no solid food has been consumed for at least about 2 hours, up to about 30 minutes before further solid food is ingested.

[0026] As used herein, the term "pharmaceutical composition" refers to a composition comprising a compound described herein formulated with a pharmaceutically acceptable excipient. The pharmaceutical composition can be formulated, for example, for oral administration in unit dosage form (e.g., tablets, capsules, caplets, gelcaps, suspensions, solutions, or syrups), for topical administration (e.g., as creams, gels, lotions, or ointments), for intravenous administration (e.g., as a sterile solution in a solvent system suitable for intravenous use, free of particulate embolisms), or in any other pharmaceutically acceptable formulation.

[0027] As used herein, the term “pharmaceutically acceptable salt” means any pharmaceutically acceptable salt of sepiapterin. For example, pharmaceutically acceptable salts of sepiapterin include those that fall within reasonable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, or allergic reactions, and have a reasonable benefit / risk ratio. Pharmacologically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977, and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. PH Stahl and CG Wermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds described herein, or separately by reacting the free base group with a suitable organic acid.

[0028] In many cases, compounds are prepared or used as pharmaceutically acceptable salts, which are prepared as addition products of pharmaceutically acceptable acids. Suitable pharmaceutically acceptable acids and methods for preparing suitable salts are well known in the art. Salts may be prepared from pharmaceutically acceptable, non-toxic acids, including inorganic and organic acids.

[0029] Typical acid addition salts include acetate, adipate, alginate, ascorbate, aspatate, benzenesulfonate, benzoate, besylate, bisulfate, borate, butylate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, gentisate, glucoheptate, glycerophosphate, glycolate, hemisulfate, heptonate, hexanate, hydrobromide, hydrochloride, hydroiodide, and 2 This includes hydroxyethanesulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectinates, parsulfates, 3-phenylpropionates, phosphates, picates, pivalates, propionates, stearates, succinates, sulfates, tarates, thiocyanates, toluenesulfonates, undecanoates, and valverate salts.

[0030] As used herein, the terms “subject” or “patient” mean any organism to which a compound or composition according to the present invention may be administered, for example, for experimental, diagnostic, preventive, and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may be a human or animal that requires treatment, is receiving treatment, may receive treatment in the future, or is being cared for by a specialist trained for a particular disease or condition.

[0031] As used herein, the term “BH4-related disease” refers to any disease or disorder for which therapeutic benefits can be obtained from the regulation of BH4 levels or activity. BH4-related disorders include, but are not limited to, primary BH4 deficiency, GTPCH deficiency, 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency, DHPR deficiency, sepiapterin reductase deficiency, dopamine-responsive dystonia, Segawa syndrome, tyrosine hydroxylase deficiency, phenylketonuria, DNAJC12 deficiency, Parkinson's disease, depression due to Parkinson's disease, impulsivity in patients with Parkinson's disease, major depressive disorder, autism spectrum disorder, ADHD, schizophrenia, bipolar disorder, cerebral ischemia, restless legs syndrome, obsessive-compulsive disorder, anxiety disorders, aggression in Alzheimer's disease, cerebrovascular disorders, convulsions after subarachnoid hemorrhage, myocarditis, coronary spasms, cardiac hypertrophy, arteriosclerosis, hypertension, thrombosis, infections, endotoxic shock, cirrhosis, hypertrophic pyloric stenosis, gastric mucosal injury, pulmonary hypertension, renal dysfunction, impotence, or hypoglycemia. In some embodiments of any of the above-described aspects, the BH4-related disorder is phenylketonuria, BH4 deficiency (e.g., primary BH4 deficiency), CNS disorder (e.g., Segawa syndrome, depression, schizophrenia, autism, or Parkinson's disease), or gastrointestinal motility disorder (e.g., gastroparesis and primary and secondary esophageal motility disorders). In some embodiments, the BH4-related disorder is BH4 deficiency or phenylketonuria.

[0032] As used herein, the terms “to treat,” “treated,” or “to treat” mean both therapeutic actions and preventive or inhibitory measures, the purpose of which is to suppress or slow (reduce) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desirable clinical outcome. Beneficial or desirable clinical outcomes include, but are not limited to, relief of symptoms, reduction in the severity of a condition, disorder, or disease, stabilization (i.e., non-worsening) of a condition, disorder, or disease, onset or delay of progression of a condition, disorder, or disease, improvement (partial or overall) of a condition, disorder, or disease, whether detectable or undetectable, improvement of at least one measurable physical parameter, which does not necessarily have to be identifiable by the patient, or improvement or enhancement of a condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes extending survival compared to the expected survival without treatment. [Brief explanation of the drawing]

[0033] [Figure 1] Figure 1 is a graph showing the mean plasma concentration of BH4 over time for subjects undergoing feeding and fasting. [Figure 2] Figure 2 is a graph showing the mean plasma concentrations of sepiapterin over time in feeding and fasting subjects. [Modes for carrying out the invention]

[0034] The inventors discovered that administering sepiapterin or a pharmaceutically acceptable salt thereof with food unexpectedly increased BH4 exposure in the subject's plasma. Therefore, the present invention features a composition comprising sepiapterin or a pharmaceutically acceptable salt thereof, and a method for treating BH4-related diseases. This composition and method can result in increased plasma exposure to BH4.

[0035] Sepiaapterin Sepiapterin passes into the cell and is converted to 7,8-dihydrobiopterin by sepiapterin reductase. 7,8-dihydrobiopterin is then converted to BH4 by reduction by dihydrofolate reductase.

[0036] While not bound by theory, administering sepiapterin with food may lead to increased plasma exposure to BH4, for example, by reducing the absorption rate of sepiapterin. When administered sepiapterin is rapidly absorbed (for example, by being administered on an empty stomach), intracellular sepiapterin reductase and / or dihydrofolate reductase are activated. max The reaction rate exceeds the V threshold of the substrate saturation of sepiapterin reductase and / or dihydrofolate reductase, resulting in saturation. As a result, at least a portion of the administered sepiapterin may leave the cell without being reduced to 7,8-dihydrobiopterin and subsequently to BH4. This excess sepiapterin may be excreted without being converted to BH4, resulting in a decrease or prolongation of the absorption rate of sepiapterin, and the reaction rate may be affected by the V threshold of the substrate saturation of sepiapterin reductase and / or dihydrofolate reductase. max Plasma BH4 levels are lower compared to administration with food, resulting in results below, the same as, or slightly above the specified threshold. When sepiapterin is administered with food, unexpectedly, the maximum plasma concentration (Cmax) and time from zero to final concentration (AUC) of BH4 are higher compared to administration without food. 0-last The degree of exposure increases as measured by the area under the concentration-time curve up to ).

[0037] Sepiapterin has the following structure. JPEG0007855033000001.jpg3044

[0038] Sepiapterin or a pharmaceutically acceptable salt thereof may be incorporated into the pharmaceutical composition. In some embodiments, the pharmaceutical composition of the present invention contains 20-30% of sepiapterin or a salt thereof by total weight, for example, 20%, 22%, 25%, 27%, or 30%. In some embodiments, the pharmaceutical composition contains more than 20% of sepiapterin by total weight, for example, 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%. In some embodiments, the pharmaceutical composition contains less than 20% of sepiapterin by total weight, for example, less than 20%, less than 15%, less than 10%, or less than 5%.

[0039] In some embodiments, the present invention features a pharmaceutical composition comprising sepiapterin or a salt thereof and an antioxidant in an amount less than 10% (e.g., 9%, 7%, 5%, 3%, 1%, 0.5%, 0.25%, or 0.1%) of the total weight. The antioxidant may be ascorbic acid. In some embodiments, the ratio of sepiapterin or a pharmaceutically acceptable salt thereof to the antioxidant is 1:1, for example, 2:1, 5:1, 7:1, or 10:1. The pharmaceutical composition may contain sepiapterin or a pharmaceutically acceptable salt thereof in an amount of 20-30% of the total weight, for example, 20%, 22%, 25%, 27%, or 30%. The pharmaceutical composition may further contain a dispersant, for example, croscamerose sodium. The pharmaceutical composition may contain a dispersant in an amount of 0.1-1.5% of the total weight, for example, 0.1%, 0.5%, 1%, or 1.5%. In some embodiments, the pharmaceutical composition comprises at least one anticaking agent, such as colloidal silicon dioxide or microcrystalline cellulose. The pharmaceutical composition may contain the anticaking agent in an amount of 65-75% of the total weight, for example, 65%, 67%, 70%, 73%, or 75%. In some embodiments, the pharmaceutical composition comprises both colloidal silicon dioxide and microcrystalline cellulose. In some embodiments, the pharmaceutical composition comprises 60-65% microcrystalline cellulose and 5-7% colloidal silicon dioxide by weight. In some embodiments, the crystalline form of sepiapterin is formulated as particles less than 140 μm (for example, 120 μm, 110 μm, 100 μm, 90 μm, 80 μm, 70 μm, 60 μm, 50 μm, 40 μm, 30 μm, 20 μm, 10 μm, or 5 μm). In some embodiments, the pharmaceutical composition contains less than 1% of an impurity such as lactoylpterin, for example, 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%.

[0040] In some embodiments, sepiapterin is a salt of sepiapterin, for example, a salt 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.

[0041] In some embodiments, sepiapterin or its pharmaceutically acceptable salts are in crystalline form. The crystalline form of crystalline sepiapterin-free base or salts of sepiapterin may occur as anhydrous (e.g., without any bound water or solvent or hydration or solvation), or as hydrates, partial hydrates (e.g., hemihydrate, sesquihydrate, etc.), dihydrates, trihydrates, etc., where the crystalline form binds the hydration water or solvent molecules associated with the crystalline form of sepiapterin or its salts. In one embodiment, crystalline sepiapterin occurs as a monohydrate or hemihydrate.

[0042] In some embodiments, sepiapterin exists in a crystalline form. In some embodiments, the crystalline sepiapterin is characterized by a powder X-ray diffraction pattern obtained by irradiation with CuKα X-rays having peaks represented by 2θ at at least about 9.7°, about 10.2°, and about 11.3°. In other embodiments, the crystalline sepiapterin is characterized by a powder X-ray diffraction pattern obtained by irradiation with CuKα X-rays having peaks represented by 2θ at at least about 9.7°, about 10.2°, about 11.3°, about 14.0°, about 14.6°, about 19.9°, about 22.2°, about 25.3°, and about 32.4°.

[0043] The present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient and an effective amount of sepiapterin or a pharmaceutically acceptable salt thereof.

[0044] Pharmaceutically acceptable excipients may be any of those conventionally used, and are limited only by physicochemical considerations such as solubility and route of administration. Those skilled in the art will understand that, in addition to the pharmaceutical compositions described below, sepiapterin can be formulated as inclusion complexes such as cyclodextrin inclusion complexes, or as liposomes.

[0045] The pharmaceutically acceptable excipients described herein, such as vehicles, adjuvants, excipients, or diluents, are well known to those skilled in the art and readily available. Preferably, the pharmaceutically acceptable excipients are chemically inert to sepiapterin and do not have adverse side effects or toxicity under the conditions of use.

[0046] Preparations that increase gastric and / or foregut retention time Gastric-retaining drug delivery is an approach designed to keep drug formulations in the stomach longer until drug release is complete.

[0047] Bioadhesive formulations utilize polymers that can adhere to surfaces and provide controlled release of the drug. The bioadhesive polymer may be anionic (e.g., carboxymethylcellulose, alginic acid, polyacrylic acid, pectin, carrageenan, polycarbophil, or carbomer), cationic (e.g., chitosan, polylysine, or polyblen), or nonionic (e.g., polyethylene glycol, polyvinylpyrrolidone, dextran, or hydroxypropyl methylcellulose).

[0048] High-density formulations are designed to be present in the stomach at a level lower than the pyloric sphincter, thus avoiding emptying the stomach. Suitable excipients for high-density formulations include iron powder, barium sulfate, zinc oxide, and titanium dioxide.

[0049] Bulk-forming formulations are designed to expand in the stomach to become larger than the pyloric sphincter, thus helping to avoid hunger. For example, formulations containing a drug core, a swellable hydrocolloid, and an outer semipermeable polymer are suitable for bulk-forming formulations.

[0050] Hyperporous hydrogel formulations, like bulk-forming formulations, are designed to expand beyond the pyloric sphincter in the stomach. Hyperporous hydrogel formulations may contain polymers such as croscarmellose sodium.

[0051] Floating formulations are designed to have a density lower than that of gastric juice. Floating formulations may include compositions comprising ion exchange resins, raft systems, inflatable chambers, effervescent mixtures, swellable hydrocolloids, or multi-particle systems.

[0052] Antioxidants Sepiapterin tends to oxidize rapidly when exposed to air. Therefore, the pharmaceutical composition of the present invention may contain antioxidants. The antioxidants may minimize the oxidative degradation of sepiapterin. Examples of antioxidants, but not limited to, include ascorbic acid, tocopherol, retinol, ascorbyl palmitate, N-acetylcysteine, glutathione, ethylenediaminetetraacetic acid, sodium bisulfite, sodium metabisulfite, thiourea, butylated hydroxytoluene, butylated hydroxyanisole, and 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 antioxidants.

[0053] In some embodiments, the pharmaceutical composition contains less than 10% by weight of the antioxidant, for example, less than 9% by weight, less than 8% by weight, less than 7% by weight, less than 6% by weight, less than 5% by weight, less than 4% by weight, less than 3% by weight, less than 2% by weight, less than 1% by weight, or substantially no antioxidant. In some embodiments, the pharmaceutical composition contains 2 to 9% of the antioxidant by weight, for example, 2 to 4%, 3 to 5%, 4 to 6%, 5 to 7%, 6 to 8%, or 7 to 9%. In some embodiments, the pharmaceutical composition contains 5 to 100% of the USP maximum daily dose of the 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 USP maximum daily dose of the antioxidant. In some embodiments, the ratio of sepiapterin to antioxidant is at least 1:1, for example, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1.

[0054] Dispersant In some embodiments, the pharmaceutical composition of the present invention comprises at least one dispersant. The dispersant may separate particles in the formulation, for example, by releasing its active ingredient upon contact with water. Examples of dispersants include, but are not limited to, cross-linked polyvinylpyrrolidone, carboxymethylcellulose (e.g., croscarmellose salt, e.g., croscarmellose sodium), starch (e.g., sodium starch glycolate), or alginic acid. In some embodiments, the dispersant in the pharmaceutical composition is carboxymethylcellulose, such as a pharmaceutically acceptable salt of croscarmellose. In some embodiments, the pharmaceutical composition may contain 0.1 to 1.5% (e.g., 0.1%, 0.5%, 1%, or 1.5%) of the dispersant based on the total weight. In some embodiments, the pharmaceutical composition contains less than 1.5% (e.g., less than 1%, less than 0.5%, or less than 0.1%) of the dispersant.

[0055] Anti-caking agent In some embodiments, the pharmaceutical composition of the present invention comprises at least one anticaking agent. In some embodiments, the pharmaceutical composition of the present invention comprises at least two anticaking agents. Exemplary anticaking agents include colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, talcum powder, sodium aluminosilicate, potassium aluminum silicate, calcium aluminosilicate, bentonite, aluminum silicate, stearic acid, and polydimethylsiloxane. In some embodiments, at least one anticaking agent is colloidal silicon dioxide or microcrystalline cellulose. In some embodiments, the pharmaceutical composition may contain 65-75% (e.g., 65%, 67%, 70%, 73%, or 75%) of the anticaking agent based on the total weight. 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 and 5-7% colloidal silicon dioxide by weight.

[0056] Administration vehicle In some embodiments, the pharmaceutical composition of the present invention is combined with an administration vehicle before administration (e.g., an administration vehicle having a viscosity of about 50 to 1750 centipoise (cP)). One type of suspension that can be used is a combination of glycerin and sucrose in water (e.g., MEDISCA containing 2.5% glycerin and 27% sucrose in water). (R) (Oral mix). An appropriate amount of the composition can be added to the administration vehicle mixture and stirred to suspend the composition immediately before administration.

[0057] Other suspensions may also be used as administration vehicles. Exemplary suspensions include agar, alginic acid, sodium carboxymethylcellulose, carrageenan, dextrin, gelatin, guar gum, hydroxyethylcellulose, hydroxypropylcellulose, hypermellose, methylcellulose, polyethylene glycol, povidone, tragacanth, xanthan gum, or other suspensions known in the art.

[0058] dose Sepiapterin or its pharmaceutically acceptable salts can be used in any appropriate dose. Appropriate doses and dose regimens can be determined within the scope of the prior art. Generally, treatment is initiated with a dose lower than the optimal dose. The dose is then gradually increased until the optimal effect is achieved under the circumstances. For convenience, the total daily dose can be divided and administered throughout the day if desired. With appropriate doses and appropriate administration of specific compounds, the present invention provides a wide range of responses. Typically, doses range from approximately 2.5 to approximately 150 mg / kg body weight / day of the patient being treated. For example, in the embodiment, sepiapterin, or a pharmaceutically acceptable salt thereof, can be administered once or more times a day at doses of approximately 20 mg / kg to approximately 150 mg / kg, approximately 40 mg / kg to approximately 100 mg / kg, approximately 100 mg / kg to approximately 150 mg / kg, approximately 60 mg / kg to approximately 120 mg / kg, approximately 80 mg / kg to approximately 100 mg / kg, approximately 40 mg / kg to approximately 60 mg / kg, approximately 2.5 mg / kg to approximately 20 mg / kg, approximately 2.5 mg / kg to approximately 10 mg / kg, or approximately 2.5 mg / kg to approximately 5 mg / kg of target body weight / day, in order to obtain the desired therapeutic effect.

[0059] In some embodiments, the dose is sufficient to generate levels of BH4 in the CNS (e.g., the brain), as measured, for example, in the CSF, and / or sufficient to produce a therapeutic outcome and / or response, such as an increase in serotonin or dopamine in the CNS. In some embodiments, the increase in BH4 in the CNS is measured by determining the levels of metabolites of monoamines, such as serotonin and / or dopamine (e.g., homovanillic acid or 5-hydroxyindoleacetic acid (5-HIAA)), in the CSF, where an increase in the metabolite in the CSF indicates an increase in BH4 levels in the CNS (e.g., the brain). In some embodiments, the dose is sufficient to increase the level of HB4, as measured in plasma or the organ of interest (e.g., the liver of the interest), to at least twice the level of BH4 before administration (e.g., at least 10 times, at least 20 times, at least 50 times, at least 100 times, or at least 150 times).

[0060] In some embodiments, sepiapterin or a pharmaceutically acceptable salt thereof can be formulated as a unit solid oral dosage form, such as particles. In these embodiments, each unit solid oral dosage form may contain any appropriate amount of sepiapterin or a pharmaceutically acceptable salt thereof. For example, each unit solid oral 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.

[0061] Sepiapterin or a pharmaceutically acceptable salt thereof can be used to prepare liquid formulations such as solutions, suspensions, or emulsions. Formulations suitable for oral administration may consist of (a) solid or granular forms, each containing a predetermined amount of the active ingredient, such as capsules, sachets, tablets, lozenges, and troches; (b) powders; (c) liquid formulations such as an effective amount of the compound dissolved in a diluent such as water, saline, or orange juice; (d) suspensions in a suitable liquid; and (e) suitable emulsions. Preferably, solid oral formulations such as capsules, tablets, and powders. Capsules may be of the usual hard or soft shell-like gelatin type, containing, for example, a surfactant, lubricant, and inert filler, such as lactose, sucrose, calcium phosphate, and corn starch. The tablet form may contain lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and one or more other excipients, colorants, diluents, buffers, disintegrants, wetting agents, preservatives, flavorings, and pharmacologically suitable excipients. The lozenge form may contain the active ingredient in addition to a flavor (usually sucrose and acacia or tragacanth), and similarly, pastilles may contain the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia, emulsion, or gel, and such excipients are known in the art.

[0062] Formulations suitable for oral and / or parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, bacteriostags, and solutes and suspensions that make the formulation isotonic with the recipient's blood, as well as aqueous and non-aqueous sterile suspensions, solubilizers, thickeners, stabilizers, and preservatives. The compounds of the present invention can be administered in a physiologically acceptable diluent in a sterile liquid or mixture of liquids containing pharmaceutical excipients, such as water, physiological saline, glucose aqueous solution and related sugar solutions, alcohols such as ethanol, benzyl alcohol, or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol and polyethylene alcohol, glycerol ketals such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers such as poly(ethylene glycol) 400, oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides with or without the addition of pharmaceutically acceptable surfactants such as soap or detergent, suspending agents such as pectin, carbohydrates, methylcellulose, hydroxypropyl methylcellulose, or carboxymethylcellulose, or emulsifiers, and other pharmaceutical adjuvants.

[0063] The present invention features pharmaceutical compositions of orally acceptable formulations comprising a therapeutically effective amount of sepiapterin and less than 10% of an antioxidant. In some embodiments, the pharmaceutical composition is a granular formulation dispersed in pharmaceutically acceptable excipients, for example, the composition can be mixed with water and ingested by a patient (e.g., within 5 to 10 minutes). Preferred formulations for use in the present invention are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA 22nd ed., 2010. Unless any conventional excipient is incompatible with the active ingredient, its use in the pharmaceutical compositions of the present invention is intended. Furthermore, for animal (e.g., human) administration, it will be understood that the formulation should meet sterility, pyrogenicity, general safety, and purity standards as required by the FDA Office's biological standards.

[0064] Oils that can be used in parenteral formulations include petroleum, animal, plant, and synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, petrolatum, and minerals. Fatty acids suitable for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters. Suitable soaps for use in parenteral formulations include fatty acid alkali metals, ammonium, and triethanolamine salts; suitable detergents include (A) cationic detergents (e.g., dimethyldialkylammonium halides and alkylpyridinium halides); (B) anionic detergents (e.g., alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates); (C) nonionic detergents (e.g., fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene-polypropylene copolymers); (D) amphoteric detergents (e.g., alkyl-β-aminopropionates and 2-alkyl-imidazopeake quaternary ammonium salts); and mixtures thereof.

[0065] Parenteral formulations may typically contain about 20% to about 30% by weight of sepiapterin or a pharmaceutically acceptable salt thereof in solution. Suitable preservatives and buffers may be used in such formulations. To minimize or eliminate irritation at the injection site, such compositions may contain one or more nonionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in such compositions is in the range of about 5% to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters such as sorbitan monooleate, and high molecular weight adducts of hydrophobic bases and ethylene oxide formed by the condensation of propylene oxide and propylene glycol. Parenteral formulations can be placed in unit-dose or multi-dose sealed containers such as ampoules and vials and can be stored in a lyophilized state requiring only the addition of a sterile liquid excipient for injection, such as water, immediately before use. Immediate injection solutions and suspensions can be prepared from the types of sterile powders, granules, and tablets described above.

[0066] The sepiapterin or a pharmaceutically acceptable salt thereof of the present invention can be prepared as an injectable formulation. The requirements for effective pharmaceutically acceptable excipients for injectable compositions are well known to those skilled in the art. Remington: The Science and Practice of Pharmacy, 22 nd See Edition, Lippincott Williams & Wilkins, (2012), and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JC Boylan, 2006, Marcel Dekker, New York (these are incorporated herein by reference).

[0067] Topical formulations, including those useful for transdermal drug release, are well known to those skilled in the art and are suitable for application to the skin in the context of the present invention. Compositions applied topically are typically in the form of liquids, creams, pastes, lotions, and gels. Topical administration includes application to oral mucosa, including the oral cavity, oral epithelium, palate, gingiva, and nasal mucosa. In some embodiments, the composition comprises sepiapterin, or a pharmaceutically acceptable salt thereof, and a suitable vehicle or excipient. It may also contain other components, such as an anti-irritant. The excipient may be a 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 vehicle of various components. In one embodiment, the primary vehicle is water or a substantially neutral or substantially neutralized biocompatible solvent. The liquid vehicle may contain buffers, alcohols, glycerin, mineral oils, and other materials, along with various emulsifiers or dispersants known in the art, to obtain the desired pH, invariance, and viscosity. The composition can be manufactured as a solid, such as a powder or granules. This solid can be applied directly before use, or dissolved in water or a biocompatible solvent to form a substantially neutral or substantially neutralized solution, which can then be applied to the target site. In embodiments of the present invention, the vehicle for topical application to the skin may include water, a buffer solution, various alcohols, glycols such as glycerin, lipid materials such as fatty acids, mineral oil, phosphoglycerides, collagen, gelatin, and silicone-based materials.

[0068] Sepiapterin or its salts can be prepared alone or in combination with other suitable ingredients as aerosol formulations for administration by inhalation. These aerosol formulations can be mounted on pressurized, acceptable propellants such as dichlorodifluoromethane, propane, or nitrogen. They may also be formulated as pharmaceuticals for non-pressurized formulations, such as nebulizers or atomizers.

[0069] Furthermore, the sepiapterin of the present invention or a pharmaceutically acceptable salt thereof may be mixed with various bases, such as emulsifying bases or water-soluble bases, to produce suppositories. Formulations suitable for vaginal administration may exist as pessaries, tampons, creams, gels, pastes, foams, or spray formulations, containing the active ingredient in addition to excipients known to be appropriate in the art.

[0070] Oral solid dosage form Formulations for oral use comprise particles containing the active ingredient in a mixture with non-toxic, pharmaceutically acceptable excipients, and such formulations are known to those skilled in the art (e.g., US Patent Nos.: 5,817,307, 5,824,300, 5,830,456, 5,846,526, 5,882,640, 5,910,304, 6,036,949, 6,036,949, 6,372,218, which are incorporated herein by reference). Excipients include, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starch containing potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate), granulating and disintegrating agents (e.g., cellulose derivatives containing microcrystalline cellulose, starch containing potato starch, croscarmellose sodium, alginates, or alginic acid), binders (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pre-gelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinyl pyropropylcellulose) The excipients may be lydone (or polyethylene glycol), lubricants, anti-adhesion agents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc), and anticaking agents (e.g., colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, talcum powder, sodium aluminosilicate, potassium aluminum silicate, calcium aluminosilicate, bentonite, aluminum silicate, stearic acid, or polydimethylsiloxane). Other pharmaceutically acceptable excipients may be colorants, fragrances, plasticizers, humectants, and buffers. In some embodiments, the excipients (e.g., fragrances) are packaged together with the composition.In some embodiments, excipients (e.g., fragrances) are packaged separately from the composition (e.g., combined with the composition before administration).

[0071] The solid composition of the present invention may include a coating adapted to protect the composition from undesirable chemical changes (e.g., chemical decomposition before the release of the active substance). The coating may be applied to the solid dosage form in a manner similar to that described in the Encyclopedia of Pharmaceutical Technology above.

[0072] Using the above components, powders and granules may be prepared by conventional methods, for example, using a mixer, fluidized bed apparatus, melt-solidification apparatus, rotor granulator, extrusion / spheroidization apparatus, or spray drying apparatus.

[0073] Treatment method Sepiapterin is used to treat disorders associated with decreased intracellular BH4 levels, including primary BH4 deficiency, GTPCH deficiency, 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency, DHPR deficiency, sepiapterin reductase deficiency, dopamine-responsive dystonia, Segawa syndrome, tyrosine hydroxylase deficiency, phenylketonuria, DNAJC12 deficiency, Parkinson's disease, depression associated with Parkinson's disease, impulsivity in Parkinson's disease patients, major depressive disorder, autism spectrum disorder, and ADHD. It may be useful in treating a variety of diseases associated with dysfunction of BH4-dependent metabolic pathways, including but not limited to schizophrenia, bipolar disorder, cerebral ischemia, restless legs syndrome, obsessive-compulsive disorder, anxiety disorders, aggression in Alzheimer's disease, cerebrovascular disease, convulsions after subarachnoid hemorrhage, myocarditis, coronary spasms, cardiac hypertrophy, arteriosclerosis, hypertension, thrombosis, infections, endotoxin shock, cirrhosis, hypertrophic pyloric stenosis, gastric mucosal injury, pulmonary hypertension, renal dysfunction, impotence, and hypoglycemia. Therefore, various forms of sepiapterin, or salts thereof, of the present invention can be administered to a patient in an effective amount to obtain treatment or improvement of a disease, disorder, or condition.

[0074] In some embodiments of any of the above methods, the food is a high-protein food. In some embodiments of any of the above methods, the food is a high-fat food (e.g., at least 25, 30, 40, or 50% of the calories come from fat). In some embodiments of any of the above methods, the food is a high-protein and high-fat food. In some embodiments, the food is a high-calorie food (e.g., the food contains at least 100 calories, e.g., at least 200 calories, at least 300 calories, at least 400 calories, at least 500 calories, e.g., 500-1500 or 800-1000 calories). In some embodiments of any of the above methods, the food is a meal, e.g., breakfast, lunch, or dinner.

[0075] 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 condition, type of disease being treated, previous or concurrent therapeutic interventions, patient idiopathy, and route of administration. Depending on the dose and route of administration, the preferred dose and / or number of effective doses may vary depending on the subject's response. In any case, the person responsible for administration may determine the concentration of the active ingredient in the composition and the appropriate dose for each individual subject.

[0076] In some embodiments, the patient receives 2.5 mg / kg / day, 5 mg / kg / day, 10 mg / kg / day, 20 mg / kg / day, 40 mg / kg / day, 60 mg / kg / day, or 80 mg / kg / day. The patient may receive the pharmaceutical composition containing sepiapterin once, twice, or three times daily during treatment. In some embodiments, the patient continues other current medications for BH4-related disorders (e.g., prescribed L-dopa / carbidopa, 5-HTP, melatonin, MAO inhibitors, and dopamine receptor agonists), except for BH4 supplements (if taking BH4). The patient may be permitted to take medications known to inhibit folate synthesis (e.g., methotrexate, pemetrexed, trimethrexate, etc.).

[0077] In some embodiments, patients taking BH4 discontinue BH4 administration (i.e., BH4 washout). Blood samples for Phe concentration may be obtained during the BH4 washout period 7, 5, 3, and 1 day prior to treatment with the pharmaceutical composition of the present invention, or at any point during the BH4 washout until the blood Phe concentration reaches >360 μmol / L. In some embodiments, pre-administration blood samples are tested for sepiapteprine, Phe, BH4, and tyrosine (Tyr).

[0078] Equivalents and scope Those skilled in the art will recognize many equivalents to the specific embodiments of the present invention described herein, or can verify them by routine experimentation alone. The scope of the present invention is not intended to be limited to the foregoing description, but rather as set forth in the appended claims.

[0079] Furthermore, it is understood that any particular embodiment of the present invention that constitutes prior art may be expressly excluded from one or more of the claims. Such embodiments are considered to be known to those skilled in the art and may be excluded even if the exclusion is not expressly provided herein. Any particular embodiment of the composition of the present invention (e.g., any compound, any production method, any use) may be excluded from one or more claims for any reason, whether or not it relates to the existence of prior art. [Examples]

[0080] Example 1. Evaluation of the effects of food on sepiapterin administration Methods: Subjects were orally administered sepiapterin (10 mg / kg) twice a week, once in a fasted state and once in a fed state. On day 8, starting 30 minutes before the second oral administration of sepiapterin, subjects were given a standard high-fat (approximately 50 percent of the total calories in the meal) and high-calorie (approximately 800-1000 calories) meal.

[0081] Sampling for PK analysis was performed before dosing on Day 1 and Day 8 (within 30 minutes before dosing), and at 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours after dosing on Day 1 and Day 8.

[0082] The blood concentrations of sepiapterin and BH4 were analyzed in the MNG laboratory.

[0083] The cerebrospinal fluid (CSF) of the selected subjects was collected by lumbar puncture approximately 30 minutes after the time point of the maximum observed plasma BH4 concentration (Tmax) determined by blood analysis on Day 1 (before dosing) and Day 7 (i.e., after daily dosing for 7 days).

[0084] The cerebrospinal fluid (CSF) was analyzed in the MNG laboratory. Descriptive statistics are provided to characterize the changes in neurotransmitter metabolism between the sample results on Day 1 and Day 7.

[0085] Results: As shown in Tables 1 and 2 and Figure 1 below, surprisingly, the Cmax of BH4 in plasma was much higher in subjects who had eaten before dosing compared to subjects who had fasted before dosing. Furthermore, when sepiapterin was administered in the fed state versus the fasted state, the plasma sepiapterin concentration decreased while the BH4 concentration increased (Figure 1). The geometric mean ratio of plasma sepiapterin (fasting / fed, 90% CI) was 1.29 (0.84 - 2.00) for AUC last and 1.57 (1.21 - 2.0) for C max . The corresponding ratio of plasma BH4 (90% CI) was 0.58 (0.47 - 0.71) for AUC 0-inf and 0.55 (0.45 - 0.68) for C max . The overall exposure of BH4 measured by AUC0- inf and AUC last increased 1.7-fold when sepiapterin was administered in the fed state compared to the fasted state.

Table 1

Table 2

[0086] Furthermore, as shown in Tables 3 and 4 and Figure 2 below, surprisingly, the Cmax of sepiapterin in plasma was significantly lower in subjects who had eaten before administration compared to subjects who had fasted before administration. [Table 3] [Table 4]

[0087] Example 2. Comparison of adverse events in feeding and fasting subjects. Methods: Twelve subjects were administered a single dose of sepiapterin (10 mg / kg) under fasting conditions, followed by administration under feeding conditions 7 days later. Standard definitions of adverse events (AEs) were used. All-factor AEs occurred at any time, and treatment-related adverse events (TEAEs) occurred during or after administration of the study treatment. TEAEs related to the study drug were based on the investigator's opinion. Serious AEs were defined as life-threatening or resulting in death, hospitalization, or extension of existing hospitalization, or resulting in persistent or significant impairment / inability to perform normal daily living activities, or substantial collapse, or congenital / birth defects.

[0088] Results: Surprisingly, as shown in Table 5, when sepiapterin was administered to feeding subjects, the incidence of adverse events was reduced compared to fasting subjects. [Table 5]

[0089] Example 3. Determination of CSF levels of neurotransmitters after administration of sepiapterin. Methods: CSF samples from subjects administered sepiapterin 60 mg / kg or placebo were analyzed. The analytes measured were sepiapterin, BH4, BH2, homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (HIAA). T1 and 7 T2 levels were measured for these analytes. maxDescriptive statistics (n, mean, SD, median, minimum, maximum) for + / - 30 minutes, and the change from baseline on day 7 were determined for each analyte.

[0090] Results: Sepiapterin was administered to BH4 T on day 1 or day 7. max No BH4 was detected in CSF at + / - 30 minutes. BH4 CSF concentration increased by 4.102 ng / ml in the sepiapterin group, but remained unchanged in the placebo group (change from baseline: 0.010 ng / ml). Similarly, BH2 CSF concentration increased by 1.368 ng / ml in the sepiapterin group, but remained unchanged in the placebo group (change from baseline: 0.020 ng / ml).

[0091] Changes in neurotransmitter concentrations from day 1 also differed between the sepiapterin-treated group and the placebo-treated group. HVA concentration increased by 1.378 ng / ml in the sepiapterin-treated group, while it decreased by 0.630 ng / ml in the placebo-treated group. 5-HIAA concentration decreased in both groups, but only slightly less in the sepiapterin-treated group than in the placebo-treated group (-1.142 ng / mL vs. -2.440 ng / mL).

[0092] Notably, the pre-administration HVA (10.02 ng / ml) and 5-HIAA (3.69 ng / ml) concentrations in one subject were approximately one-third of those observed in other subjects (HVA: 22.45 ng / ml to 44.72 ng / ml, 5-HIAA: 9.49 ng / ml to 19.70 ng / ml). However, by day 7, the HVA (29.93 ng / ml) and 5-HIAA (9.53 ng / ml) concentrations in this subject approached the range observed in other sepiapterin-treated groups (HVA: 30.64 ng / ml to 43.19 ng / ml, 5-HIAA: 10.10 ng / ml to 21.59 ng / ml). Both the HVA and 5-HIAA concentrations in this subject on day 7 were higher than the average concentrations in the placebo-treated subjects on day 7. The data are summarized in Table 6 below. [Table 6]

[0093] Example 4. Pharmacokinetic analysis of multiple doses of sepiapterin Methods: Eight feeding subjects were randomized into three cohorts and administered either sepiapterin or placebo once daily for 7 days in a 6:2 ratio. The highest dose of sepiapterin was administered using a sentinel dosing strategy.

[0094] Results: Plasma time-dependent concentrations of sepiapterin and BH4 were similar at 1 day and 7 days after sepiapterin administration, and no drug accumulation was observed. Pharmacokinetic data are shown in Table 7 below. [Table 7]

[0095] Other embodiments This specification is provided in conjunction with a detailed description, but it will be understood that the above description is intended to be illustrative and not to limit the scope of the disclosure herein as defined by the appended claims. Other embodiments, advantages, and modifications are within the scope of the following claims.

Claims

1. A composition comprising sepiapterin or a pharmaceutically acceptable salt thereof for use in the treatment of a tetrahydrobiopterin (BH4)-related disorder selected from BH4 deficiency or phenylketonuria in a subject requiring such treatment, wherein the treatment comprises orally administering an effective amount of the composition to the subject once daily with food, the administration to the subject being performed less than 30 minutes before or at the same time as food intake.

2. The composition for use according to claim 1, wherein the effective amount is sufficient to produce a BH4 concentration of at least 50 ng / mL in the target plasma within 10 hours of administration.

3. The composition for use according to claim 1 or 2, wherein the effective amount is 2.5 mg / kg to 100 mg / kg.

4. The composition for use according to claim 3, wherein the effective amount is 2.5 mg / kg to 20 mg / kg.

5. The composition for use according to claim 3, wherein the effective amount is 40 mg / kg to 60 mg / kg.

6. The composition for use according to claim 3, wherein the effective amount is 60 mg / kg.

7. A composition for use according to any one of claims 1 to 6, wherein administration to the subject is carried out simultaneously with food.

8. The composition for use according to any one of claims 1 to 7, wherein the effective amount results in an increase in the maximum plasma concentration (Cmax) of BH4 compared to administration without food.

9. The composition for use according to any one of claims 1 to 8, wherein the effective amount results in an increase in the area under the concentration-time curve from time zero to final concentration (AUC0-last) of BH4 compared to administration without food.

10. The composition for use according to any one of claims 1 to 9, wherein the food contains at least 300 calories.

11. The composition for use according to any one of claims 1 to 10, wherein the food is a high-fat food and at least 25% of the calories come from fat.

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