Treatment of Hyperammonemia

Abstract

The present invention relates to a compound of formula (I): TIFF2024516615000012.tif52161 or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

Description

[Technical field]

[0001] FIELD OF THEINVENTION The present application relates to pharmaceutical compounds and compositions useful for the prevention or treatment of hyperammonemia, particularly drug-induced hyperammonemia.

[0002] Technology background Hyperammonemia is a metabolic disorder characterized by high levels of the nitrogen-containing compound ammonia in the blood. Ammonia is an important source of nitrogen and is necessary for amino acid synthesis. Ammonia is also required for normal acid-base balance. However, when present in high concentrations, ammonia is toxic.

[0003] Ammonia is normally produced in the large and small intestine, from where it is transported to the liver and converted to the water-soluble compound urea via the urea cycle, and then excreted by the kidneys.

[0004] The consequences of hyperammonemia are varied and can be severe. Ammonia is a potent neurotoxin and, once in the brain, causes neurological damage that can lead to convulsions, ataxia, stroke-like lesions, coma, psychosis, loss of vision, acute encephalopathy and cerebral edema. Other symptoms are also observed in individuals with hyperammonemia, such as vomiting, respiratory alkalosis, hypothermia and liver damage leading to death.

[0005] Hyperammonemia must be recognized early and treated immediately to avoid potentially fatal complications such as cerebral edema or cerebral herniation.

[0006] Currently, management of hyperammonemia involves a combination of measures to lower the ammonia concentration in the blood. Typically, patients are placed on a low protein diet in combination with medications aimed at promoting nitrogen excretion, such as sodium phenylbutyrate, sodium benzoate, and glycerol phenylbutyrate (Enns et al., (2007) New Engl J Med., 356:2282-2292).

[0007] However, these drugs must be administered several times a day and are associated with many side effects, in particular nausea, vomiting, irritability and anorexia. Furthermore, the need to calculate the dose makes overdosing prone to occur, which can lead to severe metabolic side effects and death.

[0008] Thus, there remains a need for effective alternative treatments and prevention of hyperammonemia.

[0009] Summary of the Invention The present invention stems from the inventors' unexpected discovery that stiripentol attenuates methionine sulfoximine (MSO)-induced hyperammonemia and methionine sulfoximine (MSO)-induced hyperammonemia-associated seizures.

[0010] Thus, the present invention provides a compound of formula (I): [ka] [In the formula, n represents 1 or 2; A1, A2 and A3 may be the same or different and represent a hydrogen atom, a halogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; R1, R2, and R3 each independently represent a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; -Y represents -OH, =O or -SH. or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

[0011] The invention also relates to a compound of formula (I) or a pharma- ceutically acceptable salt, hydrate or solvate thereof for the use as defined above, in combination with at least one additional compound for the prevention or treatment of hyperammonemia or of one or more symptoms associated with hyperammonemia.

[0012] The present invention also relates to a pharmaceutical composition comprising as an active substance at least one compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, optionally in association with a pharma- ceutically acceptable carrier, for use in the prevention or treatment of hyperammonemia.

[0013] The present invention also relates to a pharmaceutical composition comprising as active substance at least one compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, optionally in association with a pharma- ceutically acceptable carrier, and optionally at least one additional compound for the prevention or treatment of hyperammonemia or of one or more symptoms associated with hyperammonemia.

[0014] The present invention also relates to a pharmaceutical composition comprising as an active substance at least one compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, optionally in association with a pharma- ceutically acceptable carrier, and at least one additional compound selected from the group consisting of neuroprotectants, anti-inflammatory agents, sodium benzoate, sodium phenylbutyrate, glycerol phenylbutyrate, N-carbamyl glutamate, sodium benzoate, sodium phenylacetate, lactulose, arginine hydrochloride, L-arginine, carglumic acid, L-ornithine, L-aspartic acid, citrulline and mixtures thereof, for the prevention or treatment of hyperammonemia or of one or more symptoms associated with hyperammonemia.

[0015] The present invention also relates to a method for the prevention or treatment of hyperammonemia in an individual, in particular as a combination product for simultaneous, separate or sequential use: at least one compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, - at least one additional compound for the prevention or treatment of hyperammonemia or one or more symptoms associated with hyperammonemia; This relates to products containing

[0016] The present invention also relates to a method for preventing or treating hyperammonemia in an individual, comprising administering to said individual an effective amount of a compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

[0017] In one embodiment of the invention, the above defined method also comprises the administration of at least one additional compound for the prevention or treatment of hyperammonemia or of one or more symptoms associated with hyperammonemia.

[0018] The present invention also relates to the use of a compound of formula (I) or a pharma- ceutically acceptable salt, hydrate or solvate thereof as defined above, for the manufacture of a medicament for the prevention or treatment of hyperammonemia in an individual.

[0019] In one embodiment of the invention, the medicament as defined above comprises at least one additional compound for the prevention or treatment of hyperammonemia or for the prevention or treatment of one or more symptoms associated with hyperammonemia.

[0020] Detailed Description of the Invention As intended herein, the term "comprising" is synonymous with "including," "containing," or "encompassing," i.e., when an object "comprises" one or more features, features other than the mentioned features may also be included in the object. Conversely, the phrase "consisting of" means "constituted by," i.e., when an object "consists of" one or more features, the object cannot include features other than the mentioned features.

[0021] Compounds of formula (I) Preferably, the compound of formula (I) as defined above has the following formula (II): [ka] [In the formula, n, A1, A2, A3 and R1 are as defined above.] It is expressed as:

[0022] More preferably, the compound of formula (I) or (II) as defined above has the following formula (III): [ka] It is expressed as:

[0023] Preferred alkyl groups according to the present invention include methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl and t-butyl.

[0024] Cl, I, Br or F atoms are preferred halogen atoms according to the present invention.

[0025] French patent FR 2173691, incorporated herein by reference, describes the synthesis of stiripentol, in particular from methylenedioxy-3,4-phenyl-1-dimethyl-4,4-penten-1-one-3. From this teaching, it is within the ordinary skill of the art to synthesize other compounds of formula (I).

[0026] As will be apparent to those skilled in the art, formulas (I), (II) and (III) as defined above represent any of the various stereoisomers encompassed by these formulas or mixtures thereof, in particular racemic mixtures thereof.

[0027] Thus, the compound of formula (III) can be a compound of formula (IIIa), a compound of formula (IIIb), or a mixture of a compound of formula (IIIa) and a compound of formula (IIIb), in particular a racemic mixture thereof. [ka] [ka]

[0028] Prevention or treatment "Hyperammonemia" refers to an increase in the concentration of ammonia in the body. Thus, hyperammonemia according to the present invention may be caused, for example, by an increase in ammonia production or a decrease in the ammonia detoxification process in the body.

[0029] Ammonia (NH3) is converted to ammonium cation (NH4 + ), and is a product of the catabolism of proteins and other nitrogenous compounds. Ammonia is converted by enzymes of the urea cycle to the less toxic substance urea, which is then excreted in the urine by the kidneys.

[0030] The urea cycle also serves as a source for the production of certain amino acids in the body (arginine, citrulline and ornithine) and includes six enzymes: carbamoylphosphate synthase I (CPS), ornithine transcarbamylase (OTC), argininosuccinate synthetase (ASS), argininosuccinate lyase (ASL), arginase and N-acetylglutamate synthase (NAGS) and two transport proteins: ornithine translocase (ORNT1) and citrine. Abnormalities in one or more of these eight proteins can result in hyperammonemia, also known as primary hyperammonemia.

[0031] Examples of urea cycle disorders that lead to primary hyperammonemia according to the present invention are: N-acetylglutamate synthase deficiency Carbamoyl phosphate synthase 1 deficiency Ornithine transcarbamylase deficiency Argininosuccinate synthase deficiency Argininosuccinate lyase deficiency Arginase 1 deficiency Hyperornithinemic-hyperammonemic-homocitrullinuria syndrome Citrulline deficiency (type 2 citrullinemia) Includes.

[0032] The hyperammonemia according to the present invention may result from inhibition of the urea cycle as a result of a disorder of intermediary metabolism characterized by reduced activity of an enzyme or protein that is not part of the urea cycle, also known as secondary hyperammonemia.

[0033] Examples of disorders that cause inhibition of the urea cycle leading to secondary hyperammonemia according to the present invention include the following: Propionic acidemia leading to glutamate deficiency Methylmalonic acidemia leading to the production of methylcitric acid ·Isovaleric acidemia 3-Hydroxy-3-methylglutaryl-CoA-lyase deficiency ATP deficiency leading to secondary CPS1 deficiency The following can be mentioned.

[0034] Further examples of secondary hyperammonemia according to the present invention are those caused by substrate deficiency, in particular: Defective fatty acid oxidation leading to CoA deficiency Disorders of the carnitine cycle leading to CoA deficiency Disorders of the pyruvate dehydrogenase complex leading to CoA deficiency Acute or chronic liver failure leading to CoA deficiency Lysinuric protein intolerance leading to citrulline, arginine and ornithine deficiencies Hyperinsulinemia-hyperammonemia syndrome leading to glutamate deficiency Deficiency of pyruvate carboxylase leading to aspartate deficiency Deficiency of pyrroline-5-carboxylate synthase leading to deficiencies of citrulline, arginine and ornithine Hyperammonemia is caused by

[0035] The hyperammonemia according to the present invention may also be associated with or caused by other pathologies, in particular liver damage, such as a pathology selected from the group consisting of acute or chronic liver failure, viral hepatitis, cirrhosis, liver dysfunction and hepatic vascular bypass resulting in reduced hepatic blood filtration, also known as acquired hyperammonemia.

[0036] The hyperammonemia according to the present invention may also be due to the ingestion of drugs or chemotherapy. This is called drug-induced hyperammonemia. Examples of drugs that are likely to cause hyperammonemia include sodium valproate, barbiturates such as primidone, glycine gel, L-asparaginase, 5-fluorouracil, sunitinib, asparaginase chemotherapy and regorafenib.

[0037] The hyperammonemia according to the present invention may result from complications of solid organ transplantation, for example lung transplantation, which is referred to as post-operative hyperammonemia.

[0038] The hyperammonemia prevented or treated according to the present invention may also be transient hyperammonemia of the newborn.

[0039] In one embodiment, the present invention relates to the prevention or treatment of disorders associated with hyperammonemia, preferably selected from the group consisting of hyperammonemia-associated convulsions and hyperammonemia-associated seizures.

[0040] individual The individual according to the present invention is preferably a human being.

[0041] Preferably, the individual according to the invention suffers from hyperammonemia, in particular drug-induced hyperammonemia.

[0042] In one embodiment of the invention, the individual is a term infant with a plasma ammonia concentration of at least 40 μM / L, at least 45 μM / L, at least 50 μM / L, at least 80 μM / L or at least 90 μM / L.

[0043] In one embodiment of the invention, the individual is a premature infant with a plasma ammonia concentration of at least 70 μM / L, at least 80 μM / L or at least 90 μM / L.

[0044] In one embodiment of the invention, the individual is a child over 1 month of age with a plasma ammonia concentration of at least 40 μM / L, at least 45 μM / L, at least 50 μM / L, at least 80 μM / L or at least 90 μM / L.

[0045] In one embodiment of the invention, the individual is an adult with a plasma ammonia concentration of at least 30 μM / L, at least 40 μM / L, at least 50 μM / L, at least 80 μM / L, at least 150 μM / L, at least 180 μM / L, at least 200 μM / L, at least 300 μM / L or at least 400 μM / L.

[0046] In one embodiment of the invention, the individual according to the invention has a urea cycle disorder.

[0047] In one embodiment of the present invention, the individual of the present invention exhibits cerebral function disorder.Preferably, the individual of the present invention exhibits at least one disorder selected from the group consisting of convulsions, convulsive seizures, confusion, agitation, impaired consciousness, lethargy, ataxia, stroke-like lesions, coma, psychosis, loss of vision, acute encephalopathy, cerebral edema and cerebral herniation.More preferably, the individual of the present invention has convulsions or convulsive seizures.

[0048] Administration Preferably, the compound of formula (I) or its pharma- ceutically acceptable salt, hydrate or solvate as defined above is administered or can be administered in a unit dose of about 50 mg to about 1500 mg, in particular about 150 mg to 300 mg, or is packaged in this unit dose. Also preferably, the compound of formula (I) or its pharma- ceutically acceptable salt, hydrate or solvate as defined above is administered or can be administered in a dosage regimen of about 5 g / day to about 45 g / day. Also preferably, the compound of formula (I) or its pharma- ceutically acceptable salt, hydrate or solvate as defined above is administered or can be administered in a dosage regimen of 50 mg / kg / day to 500 mg / kg / day, preferably 250 mg / kg / day to 350 mg / kg / d, more preferably about 300 mg / kg / d.

[0049] Preferably, the compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, the pharmaceutical composition as defined above or the medicament as defined above is in a form acceptable for oral or rectal administration.Preferably, the compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, the pharmaceutical composition as defined above or the medicament as defined above is administered or can be administered in the form of powders, tablets, capsules, sachets or suppositories.

[0050] In one embodiment, the compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, the pharmaceutical composition as defined above or the medicament as defined above is not administered in combination with another anticonvulsant or antiepileptic agent, such as, for example, sodium valproate and clobazam.

[0051] In one embodiment, the compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, the pharmaceutical composition as defined above or the medicament as defined above is not administered in conjunction with carnitine treatment.

[0052] Preferably, the additional compounds for the prevention or treatment of hyperammonemia or one or more symptoms associated with hyperammonemia according to the present invention are intended to prevent or treat hyperammonemia, in particular by reducing the production of ammonia in the blood or by improving the mechanisms of removal of ammonia from the blood.

[0053] Also preferably, the additional compound for the prevention or treatment of hyperammonemia, or the prevention or treatment of one or more symptoms associated with hyperammonemia, may be any compound intended to alleviate or treat one or more symptoms associated with hyperammonemia, such as a neuroprotectant or an anti-inflammatory agent.

[0054] Examples of symptoms associated with hyperammonemia include vomiting, anorexia, respiratory alkalosis, hypothermia, liver damage, and neurological disorders such as convulsions, seizures, confusion, agitation, impaired consciousness, lethargy, ataxia, stroke-like lesions, coma, psychosis, loss of vision, acute encephalopathy, cerebral edema, and cerebral herniation.

[0055] Preferably, the additional compound useful for the prevention or treatment of hyperammonemia is selected from the group consisting of neuroprotectants, such as N-acetylcysteine, indomethacin, propofol, minocycline and mannitol, anti-inflammatory agents, sodium benzoate, sodium phenylbutyrate, glycerol phenylbutyrate, N-carbamylglutamic acid, sodium benzoate / sodium phenylacetate, lactulose, arginine hydrochloride, L-arginine, carglumic acid, a mixture of L-ornithine and L-aspartic acid, citrulline and mixtures thereof.

[0056] The compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, the pharmaceutical composition as defined above or the medicament as defined above may also be accompanied by a therapy intended to alleviate or relieve one or more symptoms associated with hyperammonemia or to prevent or treat hyperammonemia, such as hemofiltration, intermittent hemodialysis, peritoneal dialysis, hypothermia therapy, renal replacement therapy.

[0057] The compound of formula (I) as defined above or a pharma- ceutically acceptable salt, hydrate or solvate thereof, the pharmaceutical composition as defined above or the medicament as defined above may also be combined with a low-protein diet and / or a diet in which ingested calories are replaced by sugars and lipids.

[0058] As used herein, the expressions "combined" or "in combination" or "combination product" mean that the compound of formula (I) as defined above or its pharma- ceutically acceptable salt, hydrate or solvate, in particular stiripentol, and the additional compound as defined above for the prevention or treatment of hyperammonemia or for the prevention or treatment of one or more symptoms associated with hyperammonemia may be combined in the same pharmaceutical composition or medicament and thus administered together, or may be administered separately, i.e. by separate administration routes and / or separate dosing schedules. However, when administered separately, the period during which the compound of formula (I) as defined above or its pharma- ceutically acceptable salt, hydrate or solvate exerts its pharmacological effect on an individual overlaps in whole or in part with the period during which the additional compound as defined above exerts its pharmacological effect on an individual.

[0059] The invention will now be further illustrated by the following non-limiting examples and figures. [Brief description of the drawings]

[0060] [Figure 1] Figure 1 shows the effect of intraperitoneally administered stiripentol (STP) on ammonia (expressed in μM, ordinate) after treatment of mice with 50 mg / kg methionine sulfoximine (MSO). *p<0.05: ANOVA statistical test of treated vs. control. #p<0.05: ANOVA statistical test of MSO and stiripentol vs. MSO only.

[0061] Working Example The present inventors investigated the efficacy of stiripentol against hyperammonemia and convulsions associated with hyperammonemia in a methionine sulfoximine (MSO)-induced convulsion model according to Hevor et al. (1985) Neuropathol. Appl. Neurobiol. 11: 129-139, as described below.

[0062] A. Materials and Methods 1.Animals Male CBA mice (Janvier) weighing 23-27 g were used after acclimation for at least 4 days in an animal house (t° = 22 ± 2°C; rearing conditions as follows: food: SAFE "A04", drinking water: tap water, day-night cycle: 12h / 12h (light: 7h / 19h - dark: 19h / 7h)).

[0063] Upon receipt, mice are grouped in batches of 8-10 in 1500U cages (floor area 1500 cm2) enriched with nesting material and sizzle dry (SDS), in accordance with the ethical recommendations of Directive 2010 / 63 / EU of the European Parliament (22 September 2010).

[0064] 2. Protocol On the day of the study, the mice are weighed, marked and placed in individual cages. At t=0 min, MSO (0.1 mL / 10 g) is administered intraperitoneally and the mice are closely observed for 8 h. The animals are normal for the first 4 h. Tonic and clonic seizures appear for several hours and then gradually disappear.

[0065] The parameters evaluated are the general condition of the animals according to the Irwin grid (Irwin 1968), the number of convulsions, their onset time and mortality. Animals are euthanized as soon as the end point is reached or at the end of the 8-h observation period according to the ANI07 protocol approved by the ethical committee.

[0066] In some studies, samples of blood and whole brain, including cerebellum, shall be taken for measurement of ammonia levels at the time of sacrifice, or at the time of death for animals that do not complete the study.

[0067] Blood is collected by cardiac puncture under isoflurane anesthesia (0.5-1.0 mL per mouse) to measure ammonia levels. For brain and blood collection, mice are quickly killed by decapitation. Brains are removed and frozen in liquid nitrogen. Livers are then removed and flash frozen in liquid nitrogen. Samples are stored at -80°C.

[0068] Stiripentol (0.1 mL / 10 g) is administered intraperitoneally to MSO at various times.

[0069] 3. Measurement of ammonia using the ABCAM kit Blood is collected after decapitation or by cardiac puncture under isoflurane anesthesia (flow rate 1.5 l / min, evaporator 4.3% for induction and maintenance) using an orange 25G needle (Terumo, NN2516R) and 1.3 ml S-Monovette tubes containing EDTA (purple cap, ref 41.1395.105, Sarstedt). The tubes are centrifuged at 2000 g for 10 min in a Beckman Allegra centrifuge cooled to 4° C. Plasma is collected in 1 ml microtubes (Brand, ref 780500). The microtubes are placed in a −80° C. freezer before analysis, otherwise they are kept on crushed ice.

[0070] Ammonia is measured colorimetrically using a microplate reader (Dynex, model MRX) and the ABCAM kit (ref AB83360, Ammonia Assay Kit), which contains several reagents. This kit allows the quantitative enzymatic measurement of ammonium present in a sample. During the assay, ammonium is converted to a product that reacts with the OxiRed probe for quantification at 570 nm using a microplate reader. The test is capable of detecting 1 nmol (#20 μM) of ammonium. The kit is stored at -20°C and contains the following components: - Ammonia Assay Buffer (25mL): ready to use, bring to room temperature before use; - OxiRed probe (200 μL): ready to use, warm to 37 °C to melt DMSO; - Developer solution (1 vial): Reconstitute with 220 μL Ammonia Assay Buffer; - Enzyme Mixture (1 vial): Reconstitute with 220 μL Ammonia Assay Buffer; -Converting Enzyme (1 vial): Reconstitute with 220 μL Ammonia Assay Buffer; - 10mM NH4Cl standard solution (100μL): ready to use, keep on ice during use

[0071] Calibration curve using NH4Cl standard solution -After each use -Prepare 100 μL of 1 mM NH4Cl standard solution -10mM NH4Cl standard solution 10μL+ED 90μL - Use 1 mM NH4Cl standard solution and prepare dilutions according to Table 1 below.

[0072] [Table 1]

[0073] The standard solution is 1 mM, i.e. 1 mmol in 1 L or 1 nmol in 1 μL, so 6 nmol in 6 μL. If you take 50 μL from 150 μL (6 + 144), you get 2 nmol / well.

[0074] Sample preparation -Plasma or serum can be tested directly -1 / 2, 1 / 5 and 1 / 10 dilutions - Plasma / serum volume of 5-15 μL per well

[0075] Ammonia measurement procedure - Equilibrate all standards / samples / materials to room temperature -Each measurement is performed twice -Well injection Standard solution: 50μL of diluted standard solution Sample: 2-50μL of sample, then adjust to 50μL with assay buffer - Prepare the reaction mixture according to Table 2 below

[0076] [Table 2]

[0077] -Add 50 μL of reaction mixture to standards and samples - Mix and incubate for 60 minutes at 37°C, protected from light. Measure at -570nm

[0078] Ammonia concentration calculation -Values ​​that are too high (outside the normal range) should be remeasured after dilution. - Average the two values -Subtract the "blank" value to obtain the corrected OD -Create a curve using standard solutions and obtain the equation (slope) -Calculate the amount of ammonium (nmol / well) (Sa) Sa = (corrected OD-y) / slope -Sample concentration (Sa / Sv) *Calculate D, where Sv is the sample volume, D is the dilution factor, and NH4 + Molar mass = 18.04 g / mol

[0079] 4.Product Methionine sulfoximine (Sigma, ref M5379, batches SLBN0115V, SLBP0971V) was dissolved in 0.9% NaCl.

[0080] Stiripentol (batch 176) was suspended in 5% (v / v) Tween 80 and 0.9% NaCl.

[0081] The ABCAM assay kit (ref AB83360, ammonia assay) and NH4Cl (ammonium chloride, Sigma, ref 254134, molar mass 53.49 g) are used for the determination of ammoniamia.

[0082] 5.Statistical analysis Results are expressed as mean ± MSE. Statistical tests used were as follows: - For seizure rates and mortality, Fisher's exact test will be used to determine significance between groups at a threshold of 5%. - A one-way analysis of variance is performed on the assay to determine whether the observed pharmacological effects are significant at the 5% level (SigmaPlot 3.1 software).

[0083] B. Results 1. Effect of administration of methionine sulfoximine Methionine sulfoximine (MSO) was administered intraperitoneally at various concentrations. Mice were observed for behavior and death up to 8 hours after administration. At -40 mg / kg, MSO produced moderate behavioral effects. From the 4th hour onwards, hypotonia, hypokinesia, accelerated respiration, ptosis and mydriasis were observed. At the end of the observation period (7-8 hours), 2 of 10 mice had convulsions and 1 had died. When started at -50 mg / kg, signs of toxicity appeared as early as 3 hours. Convulsions and death began 4 hours after MSO injection. 80% of mice had convulsions and mortality was 90%. At -60 mg / kg, methionine sulfoximine caused convulsions in 100% of mice and mortality in 90%. At -80 mg / kg, methionine sulfoximine caused convulsions in 100% of mice and 100% mortality.

[0084] Based on these results, a dose of 50 mg / kg methionine sulfoximine was selected to study the protective effect of stiripentol against MSO-induced injury.

[0085] 2. Specific effects of stiripentol on ammonia Stiripentol was administered intraperitoneally at 300 mg / kg without methionine sulfoximine.

[0086] It was observed that stiripentol did not alter ammonia (176±14 μM) compared to the control group (179±11 μM).

[0087] 3. Effect of stiripentol after administration of 50mg / kg MSO 2.1. Stiripentol administered immediately before MSO Stiripentol is administered immediately before methionine sulfoximine according to Hevor et al. (1985) Neuropathol. Appl. Neurobiol. 11:129-139.

[0088] Administration of 50 mg / kg methionine sulfoximine produced behavioral signs of toxicity, a 90% rate of seizures, and a 75% mortality rate.

[0089] Administration of 300 mg / kg stiripentol immediately before MSO significantly reduced the convulsion rate to 50% and the mortality rate to 25%, suggesting a protective effect.

[0090] In this study, the determination of ammonia gave the following results: In the control group not receiving methionine sulfoximine or any treatment, basal ammonia is measured at 174±18 μM. Administration of -50 mg / kg methionine sulfoximine significantly increased ammonia to 609 ± 51 μM. Administration of 300 mg / kg stiripentol immediately prior to -MSO reduced this hyperammonemia by 32%, to 417±51 μM.

[0091] 2.2. Stiripentol administered 30 minutes before MSO Stiripentol is administered intraperitoneally 30 min before methionine sulfoximine according to Cloix (2010) Epilepsia, 51:118-128.

[0092] Administration of 50 mg / kg methionine sulfoximine produced behavioral signs of toxicity, a 100% rate of seizures, and an 80% mortality rate.

[0093] Administration of 300 mg / kg stiripentol 30 min before MSO significantly reduced the convulsion rate to 45% and the mortality rate to 25%, suggesting a protective effect.

[0094] In this study, the following results were given based on the assessment of ammonia (Figure 1). In the control group not receiving -methionine sulfoximine or any treatment, ammonia was measured at 184±12 μM. Administration of -50 mg / kg methionine sulfoximine significantly increased ammonia to 763 ± 71 μM. Administration of 300 mg / kg stiripentol 30 min before -MSO significantly reduced this hyperammonemia by 41% to 448 ± 65 μM compared with the MSO-only group.

[0095] C. Conclusion In a methionine sulfoximine (MSO)-induced convulsion model, the present inventors showed that a single intraperitoneal administration of 50 mg / kg MSO caused hyperammonemia, convulsions, and death in 80-100% of mice.

[0096] We showed that stiripentol administered intraperitoneally at 300 mg / kg significantly reduced methionine sulfoximine-induced convulsions and mortality, suggesting a protective effect.

[0097] The primary mechanism of action of MSO is the irreversible inhibition of glutamine synthase, an enzyme involved in the detoxification of ammonia, especially in the brain, which leads to hyperammonemia (Ratnakumari, et al. (1985) J. Neurosci. Res. 14: 449-59). In astrocytes, glutamine synthase uses ammonium and glutamate to produce glutamine, so inhibition of its activity by MSO leads to an increase in ammonia.

[0098] Regarding ammonia, we showed that intraperitoneal administration of stiripentol at 300 mg / kg significantly reduced MSO-induced hyperammonemia.

Claims

1. For use in the prevention or treatment of hyperammonemia in an individual, a compound of formula (I): 【Chemistry 6】 [In the formula, n represents 1 or 2; A １ , A ２ and A ３ may be the same or different and represent a hydrogen atom, a halogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms, R １ , R ２ and R ３ independently represents a hydrogen atom or a linear or branched alkyl group having 1 to 4 carbon atoms; -Y represents -OH, ═O or -SH. or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

2. The compound represented by formula (I) may be represented by the following formula (II): 【Chemistry 7】 [Wherein, n, A1, A2, A3 and R １ is as defined in claim 1. The pharmaceutical composition according to claim 1, wherein

3. The compound represented by formula (I) is represented by the following formula (III): 【Chemistry 8】 The pharmaceutical composition according to claim 1, wherein

4. The pharmaceutical composition according to any one of claims 1 to 3, for preventing or treating convulsions associated with hyperammonemia.

5. The pharmaceutical composition according to any one of claims 1 to 3 for the prevention or treatment of drug-induced hyperammonemia.

6. The pharmaceutical composition of any one of claims 1 to 3, packaged in a unit dose of 50 mg to 1500 mg.

7. The pharmaceutical composition according to any one of claims 1 to 3, which is suitable for oral administration.

8. The pharmaceutical composition according to any one of claims 1 to 3, in the form of a powder, tablet, capsule, sachet or suppository.

9. 4. The pharmaceutical composition according to any one of claims 1 to 3, for use in combination with at least one additional compound for the prevention or treatment of hyperammonemia or one or more symptoms associated with hyperammonemia.

10. 4. The pharmaceutical composition of any one of claims 1 to 3, for use in combination with at least one additional compound selected from the group consisting of neuroprotectants, anti-inflammatory agents, sodium benzoate, sodium phenylbutyrate, glycerol phenylbutyrate, N-carbamyl glutamate, sodium benzoate, sodium phenylacetate, lactulose, arginine hydrochloride, L-arginine, carglumic acid, L-ornithine, L-aspartic acid, citrulline and mixtures thereof, for the prevention and treatment of hyperammonemia or the prevention or treatment of one or more symptoms associated with hyperammonemia.

11. A pharmaceutical composition comprising as active substance at least one compound of formula (I) as defined in any one of claims 1 to 3 or a salt, hydrate or solvate thereof, optionally with a pharma- ceutically acceptable carrier, and optionally at least one additional compound for the prevention or treatment of hyperammonemia or one or more symptoms associated with hyperammonemia.

12. A pharmaceutical composition comprising as an active substance at least one compound of formula (I) as defined in any one of claims 1 to 3 or a salt, hydrate or solvate thereof, optionally together with a pharma- ceutically acceptable carrier, and at least one additional compound selected from the group consisting of neuroprotectants, anti-inflammatory agents, sodium benzoate, sodium phenylbutyrate, glycerol phenylbutyrate, N-carbamyl glutamate, sodium benzoate, sodium phenylacetate, lactulose, arginine hydrochloride, L-arginine, carglumic acid, L-ornithine, L-aspartic acid, citrulline and mixtures thereof, for the prevention or treatment of hyperammonemia or the prevention or treatment of one or more symptoms associated with hyperammonemia.

13. For use in the prevention or treatment of hyperammonemia in an individual, in particular as a combination product for simultaneous, separate or sequential use; at least one compound of formula (I) as defined in any one of claims 1 to 3 or a pharma- ceutically acceptable salt, hydrate or solvate thereof; at least one additional compound for the prevention or treatment of hyperammonemia or one or more symptoms associated with hyperammonemia; Products containing:

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