Use of Alverine or its derivatives for the treatment of mitochondrial diseases or dysfunctions associated with mitochondrial complex I deficiency

Abstract

The present invention relates to the use of Alverine or one of its derivatives for treating diseases associated with mitochondrial dysfunction, in particular diseases associated with mitochondrial complex I deficiency.

Description

[Technical Field]

[0001] The present invention provides new pharmacological tools for treating mitochondrial diseases or dysfunctions, particularly those associated with mitochondrial complex I deficiency. [Background technology]

[0002] Mitochondrial diseases are chronic, long-term, mostly genetic, and often inherited conditions that occur when mitochondria are unable to produce enough energy for the body to function normally. Mitochondrial diseases can be present at birth but can occur at any age. It is estimated that 1 in 4,300 people has a mitochondrial disease (Gorman et al., 2015).

[0003] Mitochondrial diseases can affect almost any part of the body, including cells of the brain, nerves, muscles, kidneys, heart, liver, eyes, ears, or pancreas. The symptoms of mitochondrial diseases depend on which cells of the body are affected. Patient symptoms can range from mild to severe, affect one or more organs, and can occur at any age. Symptoms of mitochondrial diseases can include: ·Poor growth Muscle weakness, muscle pain, hypotonia, exercise intolerance Vision and / or hearing problems Learning disabilities, developmental delays, mental retardation Autism and autistic-like traits Heart, cardiac dysfunction, arrhythmia or conduction defects Liver or kidney disease Gastrointestinal disorders, difficulty swallowing, diarrhea or constipation, unexplained vomiting, cramps, reflux ·Diabetes Increased risk of infection Neurological problems, seizures, migraines, strokes Movement disorders Thyroid and / or adrenal dysfunction Breathing problems Lactic acidosis (accumulation of lactic acid) Dementia.

[0004] Mitochondrial dysfunction can also occur when mitochondria do not function normally, possibly due to another disease or condition. Many conditions can cause secondary mitochondrial dysfunction, which can affect other diseases such as Alzheimer's disease or Parkinson's disease, muscular dystrophy, Lou Gehrig's disease, diabetes, and cancer. Individuals with secondary mitochondrial dysfunction do not have a primary genetic mitochondrial disease but have similar symptoms. In addition, some medications can damage mitochondria.

[0005] Mitochondrial complex I deficiency is the most common defect, accounting for over 30% of mitochondrial diseases. Among these, the two most frequent clinical phenotypes associated with complex I deficiency are the life-threatening Leigh syndrome and milder phenotypes such as Leber's hereditary optic neuropathy (LHON). MELAS syndrome is also considered a common disorder resulting from mutations in the mitochondrial genome and is associated with severely reduced mitochondrial complex I activity.

[0006] Complex I is composed of at least 44 subunits, seven of which, i.e., ND1-ND6 and ND4L, are encoded by mitochondrial genes, while the others are nuclear-encoded. As a result, the clinical and molecular features associated with hereditary complex I deficiency are highly variable. Among these complex I subunits, mutations targeting the NDUFV1 gene have been shown to contribute to a severe neurological phenotype (Schuelke et al., 1999). Mutations affecting the NDUFS8 subunit have been associated with Leigh syndrome (Procaccio et al., 2004), and mutations targeting the mitochondrial DNA-encoded ND3 subunit have been reported in Leigh syndrome or LHON (Sarzi et al., 2007; Wang et al., 2009). Furthermore, mutations affecting the mitochondrial DNA-encoded ND6 subunit have been reported in LHON (John et al., 1992).

[0007] In addition, complex I deficiency has been identified in secondary mitochondrial dysfunction associated with age-related diseases such as Parkinson's disease.

[0008] Even though the majority of mitochondrial diseases are genetic in origin, gene therapy appears difficult to implement due to the diversity and complexity of the diseases.

[0009] The current goal of treatment is to improve symptoms and slow the progression of the disease or dysfunction, with the following recommendations, for example: Use of vitamin therapy Energy saving ·Pace activities Maintaining ambient temperature Avoid exposure to illness · Ensure adequate nutrition and hydration.

[0010] However, there remains a need to find new therapeutic pharmacological approaches to treat these types of dysfunctions or diseases. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] "Remington's Pharmaceutical Sciences" by E.W. Martin [Non-patent document 2] "Molecular Cloning: A Laboratory Manual", 4th edition (Sambrook, 2012) [Non-patent document 3] "Oligonucleotide Synthesis" (Gait, 1984) [Non-patent document 4] “Culture of Animal Cells” (Freshney, 2010) [Non-Patent Document 5] "Methods in Enzymology", "Handbook of Experimental Immunology" (Weir, 1997) [Non-patent document 6] "Gene Transfer Vectors for Mammalian Cells" (Miller and Calos, 1987) [Non-Patent Document 7] "Short Protocols in Molecular Biology" (Ausubel, 2002) [Non-patent document 8] “Polymerase Chain Reaction / Principles, Applications and Troubleshooting” (Babar, 2011) [Non-Patent Document 9] "Current Protocols in Immunology" (Coligan, 2002) Summary of the Invention [Means for solving the problem]

[0012] The present inventors have shown that alverine (ALV), a pharmacological compound primarily known as a smooth muscle relaxant used in functional gastrointestinal disorders, is a promising candidate for treating mitochondrial dysfunction or disease, particularly defects associated with complex I deficiency. This application demonstrates that it is effective in a wide range of mitochondrial diseases while exhibiting low toxicity.

[0013] definition The following definitions represent the meanings commonly used in the context of the present invention and should be considered unless another definition is explicitly stated.

[0014] In the framework of the present invention, the articles "a" and "an" are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means at least one element, i.e., one or more elements.

[0015] When used in reference to a measurable value such as amount, duration, etc., the terms "nearly," "about," or "approximately" should be understood to encompass a variation of ±20% or ±10%, preferably ±5%, more preferably ±1%, and even more preferably ±0.1% from the specified value.

[0016] Intervals / Ranges: Throughout this disclosure, various aspects of the invention may be presented in the form of interval values ​​(range format). It should be understood that the description of values ​​in interval form is merely for convenience and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values ​​within that range. For example, description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numerical values ​​within the range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0017] "Isolated" means altered or removed from its natural environment or condition. For example, an isolated nucleic acid or peptide is one that has been extracted from the natural environment in which it is normally found, e.g., in a plant or living animal. For example, a nucleic acid or peptide that occurs naturally in a living animal is not an isolated nucleic acid or peptide within the meaning of the present invention, whereas the same nucleic acid or peptide, partially or completely separated from other components present in its natural environment, is itself "isolated" within the meaning of the present invention. An isolated nucleic acid or protein can exist in a substantially purified form, or can exist in a non-native environment, such as, for example, a host cell.

[0018] The term "abnormal," when used in the context of organisms, tissues, cells, or components thereof, refers to those organisms, tissues, cells, or components thereof that differ in at least one observable or detectable characteristic (e.g., age, treatment, time of day, etc.) from those organisms, tissues, cells, or components thereof that exhibit "normal" (expected) respective characteristics. A characteristic may be normal or expected for one cell or tissue type and abnormal for a different cell or tissue type.

[0019] The terms "patient," "subject," "individual," and the like are used interchangeably herein and refer to any animal, or cells thereof, whether in vitro or in situ, following the methods described herein. In certain non-limiting embodiments, the patient, subject, or individual is an animal, preferably a mammal, more preferably a human. It may also be a mouse, rat, pig, dog, or non-human primate (NHP), such as a macaque monkey.

[0020] In the sense of the present invention, a "disease" or "pathology" is a state of health of an animal in which its homeostasis is adversely affected and which will continue to deteriorate if the disease is not treated. Conversely, in the sense of the present invention, a "disorder" or "dysfunction" is a state of health in which the animal is able to maintain homeostasis, but in which the animal's state of health is less favorable than it would be in the absence of the disorder. If left untreated, an injury does not necessarily lead to a deterioration in the animal's state of health over time.

[0021] A disease or disorder is "alleviated" ("reduced") or "improved" ("ameliorated") if the severity of the symptoms of the disease or disorder, the frequency with which such symptoms are experienced by the subject, or both, is reduced. This also includes the elimination of disease progression, i.e., the halting of progression of the disease or disorder. A disease or disorder is "cured" ("recovered") if the severity of the symptoms of the disease or disorder, the frequency with which such symptoms are experienced by the patient, or both, are eliminated.

[0022] In the context of the present invention, a "therapeutic" treatment is a treatment administered to a subject who exhibits symptoms (signs) of pathology with the purpose of reducing or eliminating these symptoms. As used herein, "treatment of a disease or disorder" means reducing the frequency or severity of at least one sign or symptom of the disease or disorder experienced by the subject. A treatment is said to be prophylactic when administered to prevent the onset, spread, or worsening of a disease, particularly when the subject does not have or does not yet have symptoms of the disease and / or the disease has not been diagnosed.

[0023] As used herein, "treating a disease or disorder" means reducing the frequency or severity of at least one sign or symptom of the disease or disorder experienced by a subject. Disease and disorder are used interchangeably herein in the context of treatment.

[0024] In the sense of the present invention, an "effective amount" or "effective dose" of a compound is an amount of the compound that is sufficient to provide a beneficial effect to the subject to which the compound is administered. The phrase "therapeutically effective amount" or "therapeutically effective amount" refers to an amount that is sufficient or effective to prevent or treat (in other words, delay or prevent the onset, prevent, inhibit, reduce or reverse the progression) a disease or disorder, including alleviating the symptoms of said disease or disorder. DETAILED DESCRIPTION OF THE INVENTION

[0025] The present invention relates to the use of alverine (ALV) or one of its derivatives, such as 4-hydroxyalverine, the first metabolite in the alverine degradation pathway, for treating diseases associated with mitochondrial dysfunction.

[0026] More particularly, according to a first aspect, the present invention therefore relates to a pharmaceutical composition comprising at least alverine (ALV) or one of its derivatives, such as 4-hydroxyalverine, for use in the treatment of diseases associated with mitochondrial dysfunction.

[0027] In other words, a composition comprising alverine (ALV) or one of its derivatives, such as 4-hydroxyalverine, is used to prepare a medicament intended for the treatment of diseases associated with mitochondrial dysfunction.

[0028] Thus, the present invention relates to a method for treating a disease associated with mitochondrial dysfunction, comprising administering to a subject in need thereof an effective dosage of a composition comprising alverine (ALV) or one of its derivatives, such as 4-hydroxyalverine.

[0029] Alverine (known as ALV), also known as N-ethyl-3-phenyl-N-(3-phenylpropyl)propan-1-amine or spasmaverine or dipropyline or cestrone or fenpropamine or alverina or N-ethyl-3,3'-diphenyldipropylamine or alverinum or fenopropamine or prophenyl, is a tertiary amine with one ethyl group and two 3-phenylprop-1-yl groups attached to the nitrogen. Its CAS number is 150-59-4 and it has the following formula:

[0030] [ka]

[0031] It is generally in the form of a white powder that has high solubility in, for example, alcohol or chloroform.

[0032] Alverine is a drug used as a smooth muscle relaxant to help treat functional gastrointestinal motility disorders.For example, it is sold under the trade name DOLOSPASMYL or METEOSPASMYL, which correspond to capsules containing 60 mg.Alverine is in the form of citrate and formulated with simethicone.In this context, the recommended daily dose is 2 or 3 capsules, i.e., 120 or 180 mg.

[0033] Also included in the present invention are derivatives of alverine that have the same biological activity, for example in mitochondrial complex I activity or mitochondrial respiration, especially as reported in the examples.

[0034] The term "alverine derivatives" encompasses derivatives and metabolites, as well as esters and pharmaceutically acceptable salts for preparing pharmaceutical compositions. Derivatives are compounds obtained from another compound (typically a precursor with a similar chemical structure) after transformation of the latter. Derivatives can differ in one or more atoms or functional groups. Metabolites are intermediate stable compounds or compounds resulting from the biochemical transformation of the original molecule by metabolism.

[0035] According to the invention, "alverine derivatives" means in particular mono- or polyhydroxylated derivatives on the phenyl nucleus and mono- or polyhydroxylated or mono- or polycarboxylated rings on the aliphatic chain.

[0036] The term "pharmaceutically acceptable salts" refers to addition salts of Alverine, which can be obtained by reacting this compound with inorganic or organic acids according to methods known per se. Acids that can be used for this purpose include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, 4-toluenesulfonic acid, methanesulfonic acid, cyclohexylsulfamic acid, oxalic acid, succinic acid, formic acid, fumaric acid, maleic acid, citric acid, aspartic acid, cinnamic acid, lactic acid, and glutamic acid. N-acetyl-aspartic acid, N-acetyl-glutamic acid, ascorbic acid, malic acid, benzoic acid, nicotinic acid, and acetic acid. Citric acid and Alverine tartrate are widely used in pharmaceutical antispasmodics.

[0037] Esters on the hydroxy functional group can include carboxylic acid esters having 1 to 6 carbon atoms.

[0038] Examples of such derivatives include:

[0039] Alverine citrate (CAS number: 5560-59-8) of the following formula:

[0040] [ka]

[0041] Alverine-d5 citrate (CAS number: 1215327-00-6) of the following formula:

[0042] [ka]

[0043] 4-Hydroxyalverine (or parahydroxyalverine; CAS number: 142047-94-7) or 4-hydroxyalverine HCl (hydrochloride) of the following formula:

[0044] [ka]

[0045] 4-hydroxyalverine-d5 (CAS number: 1216415-67-6) of the following formula:

[0046] [ka]

[0047] 4-hydroxyalverine glucuronide of the following formula:

[0048] [ka]

[0049] N-Desethylalverine HCl (CAS number: 93948-20-0) of the following formula:

[0050] [ka]

[0051] Of particular interest are alverine citrate and 4-hydroxyalverine.

[0052] The above compounds, including alverine, can be further modified to increase their stability, their bioavailability, and / or their ability to reach target tissues, particularly mitochondria.

[0053] As known to those skilled in the art, the compounds, in particular Alverine, can be present in the composition in naked form (free) or contained in delivery systems that enhance stability, targeting and / or biodisponibility, such as liposomes, or incorporated into carriers such as hydrogels, cyclodextrins, biodegradable nanocapsules, bioadhesive microspheres, vectors, or combined with cationic peptides.

[0054] The present invention also relates to a pharmaceutical composition containing at least a compound as defined above as an active ingredient, as well as to the use of this compound or composition as a pharmaceutical product or medicament.

[0055] According to certain embodiments, the pharmaceutical compositions of the present invention may comprise simethicone, particularly in connection with alverine citrate. According to other embodiments, such compositions are free of simethicone.

[0056] The present invention next provides pharmaceutical compositions comprising a compound according to the present invention. Advantageously, such compositions comprise a therapeutically effective amount of the compound and a pharmaceutically acceptable carrier. In specific embodiments, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency for use in animals and humans, or listed in the United States or European Pharmacopoeia or other generally recognized pharmacopeia. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a therapeutic agent is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like.

[0057] If desired, the compositions may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions may take the form of a solution, suspension, emulsion, sustained-release formulation, etc. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin. Such compositions contain a therapeutically effective amount of the therapeutic agent, preferably in purified form, together with a suitable amount of carrier to provide a form for proper administration to a subject.

[0058] In a preferred embodiment, the composition is formulated according to conventional procedures as a pharmaceutical composition adapted for, for example, oral administration to humans.Typically, compositions for oral administration are in the form of capsules or tablets, possibly in the form of scored tablets or effervescent tablets, and further contain excipients suitable for solid dosage forms and administration in humans.As an example, the commercially available form of Alverine is a capsule of Alverine citrate, which further contains simethicone, gelatin, glycerol and titanium dioxide.

[0059] Alternatively, the composition may be in liquid form, advantageously an aqueous composition. Any other suitable solvent may be used.

[0060] The amount of the therapeutic agent of the present invention, i.e., the compound disclosed above, effective for treating a disease can be determined by standard clinical techniques.In addition, in vivo and / or in vitro assays can be used as needed to help predict the optimal dosage range.The exact dosage to be used in the formulation also depends on the route of administration, body weight and severity of the disease, and should be determined according to the judgment of the physician and the individual circumstances of each patient.

[0061] According to certain embodiments, the compositions of the invention are in solid form, advantageously capsules or tablets, and more advantageously contain 60 mg or less of the active compound. Preferably, the compositions contain an amount of 50 mg or less, 40 mg or less, 30 mg or less, 20 mg or less, or 10 mg or less, or 5 mg or less.

[0062] According to another embodiment, the composition of the invention is in liquid form and advantageously contains less than 10 μM of active compound, or less than 3 μM, advantageously less than or equal to 1 μM, more advantageously between 100 nM and 300 nM.

[0063] Appropriate administration should allow for the delivery of a therapeutically effective amount of the therapeutic product to the target tissue, depending on the disease.

[0064] Alverine or its derivatives can be administered in a pharmaceutically acceptable form by one of the various routes known for this type of active principle.

[0065] Available routes of administration are topical (local), rectal, enteral (system-wide acting but delivered via the gastrointestinal (GI) tract), intranasal, or parenteral (systemic acting but delivered by a route other than the GI tract). In the particular case of mitochondrial disorders, the preferred route of administration of the compositions disclosed herein is enteral administration, which generally includes oral, sublingual, or buccal administration, preferably oral administration.

[0066] According to other embodiments, it may be parenterally administered, in particular intramuscularly (i.e., into the muscle) or systemically (i.e., into the circulatory system). In this context, the term "injection" (or "perfusion" or "infusion") includes intravascular, in particular intravenous (IV), intraperitoneal (IP) and intramuscular (IM) administration. Injections are usually performed using a syringe or a catheter.

[0067] According to one embodiment, the composition is administered orally, intramuscularly, intraperitoneally, subcutaneously, topically, locally or intravenously.

[0068] The pharmaceutical compositions of the present invention may be in any of the usual oral dosage forms including tablets, capsules and liquid preparations such as elixirs and suspensions containing various masking substances for color, flavor and stabilization.

[0069] According to a preferred embodiment, the composition is for oral administration. Advantageously, the composition is administered orally, that is, via the mouth.

[0070] Preferably, the compositions according to the invention are administered orally, especially in the form of capsules or tablets.

[0071] To prepare oral dosage forms, particularly capsules, according to the present invention, the active substance can be mixed with a variety of conventional materials, such as starch, calcium carbonate, lactose, sucrose, and dicalcium phosphate, to facilitate the encapsulation process. Magnesium stearate as an additive provides a useful lubricating function, if needed.

[0072] In certain cases it is advantageous to provide a form that has a controlled release, particularly a sustained release according to known galenical forms.

[0073] Likewise, the composition according to the invention is for preparing a pharmaceutical composition that can be administered by the injectable route.

[0074] For intravenous administration, the pharmaceutical compositions according to the present invention can be dissolved or suspended in a pharmaceutically acceptable sterile injectable liquid, such as sterile water, sterile organic solvent, or a mixture of these two liquids.

[0075] Other routes of administration may include, but are not limited to, subcutaneous implants, as well as oral, sublingual, transdermal, topical, intranasal, or rectal administration. Biodegradable and non-biodegradable delivery systems may also be used.

[0076] As already mentioned, the compositions according to the invention are preferably in a solid dosage form suitable for oral administration, advantageously in the form of one or more capsules or tablets, and can therefore be taken with a small amount of water before or during a main meal.

[0077] According to a preferred embodiment, the compositions according to the invention are administered daily, for example, once, twice, or three times daily, and the treatment can continue for weeks, months, years, or even lifelong.

[0078] Generally, the dosage of the therapeutic agent, i.e., Alverine or one of its derivatives, will vary depending on factors such as the subject's age, weight, height, sex, general medical condition and past medical history, etc. Typically, it is desirable to provide the patient with an individual dose of the therapeutic agent that is non-toxic and effective.

[0079] According to certain embodiments of the invention, the dosage of the composition, advantageously a daily dosage for oral intake by a human, is less than or equal to 10 mg / kg or 9, 8, 7, 6, 5, 4, 3 mg / kg, or less than or equal to 2.5, 2, 1.5 or 1 mg / kg, or less than or equal to 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3 or 0.1 mg / kg.

[0080] As already mentioned, the patient is preferably a human, in particular a neonate, infant, child, adolescent or adult, however, the therapeutic tool according to the invention may also be applicable and useful in the treatment of other animals, in particular mice, pigs, dogs or macaques.

[0081] As already mentioned, the present invention relates to the treatment of mitochondrial diseases in general, i.e. diseases associated with or caused by mitochondrial dysfunction. In the framework of this application, the term "diseases associated with mitochondrial dysfunction" is used to encompass all these conditions.

[0082] Diseases of particular interest, relating to examples showing the positive effect of Alverine or one of its derivatives on the mitochondrial respiratory chain, are mitochondrial respiratory chain disorders.

[0083] Several mitochondrial diseases are well documented in the prior art.

[0084] NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome is caused by various mutations in the mitochondrially encoded ATP6 gene, which encodes the α subunit of ATPase (OXPHOS complex V). Mutations are often heteroplasmic (coexistence of both mutant and wild-type mitochondrial DNA, mtDNA) within the same cell. Depending on both the type of mutation and the percentage of mutant mtDNA (degree of heteroplasmy), the clinical outcome is more or less severe. The ATP6m.8993T>C / G mutation is the most frequent in NARP patients and results in a severe form of NARP syndrome. FMC1 is a nuclear gene encoding a protein required for assembly of the F1 sector of ATP synthase at high temperatures (35–37°C), thereby mimicking the heteroplasmy observed in NARP patients. Indeed, when grown at the restrictive temperature (35–37°C), mitochondria from fmc1Δ mutants have far fewer assembled ATP synthase complexes than wild-type (WT) strains, although the assembled mitochondria are fully functional. This heterogeneity is also found in patients with reduced ATP synthase levels due to heteroplasmic ATP6 mutations. Thus, the fmc1Δ mutant constitutes a suitable model for these disorders, particularly the equivalent of the m.8993T>G(MR14,NARP) mutant (Schon, EA et al., 2001).

[0085] The TAZ gene encodes tafazzin, a mitochondrial transacylase that catalyzes the remodeling of immature cardiolipin to its mature composition containing a predominance of tetralinoleoyl moieties. TAZ mutations result in Barth syndrome, an X-linked disorder typically characterized by dilated cardiomyopathy (CMD) with endocardial fibroelastosis (EFE), primarily proximal skeletal myopathy, growth retardation, neutropenia, and organic aciduria, particularly excess 3-methylglutaconic acid (Barth, PG et al., 1996).

[0086] SHY1 is the yeast homolog of the human SURF1 gene. SURF1 encodes an assembly factor for mitochondrial complex IV. Mutations in SURF1 are associated with Leigh syndrome, a progressive, severe neurodegenerative disorder that begins within the first few months or years of life and can lead to early death. Affected individuals typically present with global developmental delay or regression, hypotonia, ataxia, dystonia, nystagmus, or ophthalmologic abnormalities such as optic atrophy.

[0087] SYM1 is the yeast homolog of the human MPV17 gene. MPV17 encodes an inner mitochondrial membrane protein of unknown function. MPV17 mutations cause mitochondrial DNA depletion syndrome, an autosomal recessive disorder characterized by progressive liver failure in infancy, often resulting in death within the first year of life. Surviving patients develop progressive neurological impairment, including ataxia, hypotonia, dystonia, and psychomotor regression (Spinazzola, A. et al., 2006).

[0088] According to a particular embodiment, the disease to be treated in the framework of the present invention is associated with or caused by a genetic defect in at least one of the following genes: MTTL1, ATP6, TAZ, SURF1, POLG, MPV17, OPA1, COA6, ND6 and BCS1L.

[0089] Of particular interest is the treatment of diseases selected from the group consisting of MELAS syndrome, maternally inherited myopathies and cardiomyopathies, NARP syndrome, Leigh syndrome, Barth syndrome, mitochondrial DNA depletion syndrome 4A (Alpers type), mitochondrial DNA depletion syndrome 4B (MNGIE type), mitochondrial recessive ataxia syndrome, sensory ataxic neuropathy and ophthalmoplegia, spinocerebellar ataxia with epilepsy, progressive external ophthalmoplegia, mitochondrial DNA depletion syndrome-6, Navajo neuropathy, Behr syndrome, mitochondrial DNA depletion syndrome 14, infantile cardioencephalomyopathy due to cytochrome c oxidase deficiency (COA6 mutation), mitochondrial complex III deficiency nuclear type 1, GRACILE syndrome, and Bjornstad syndrome.

[0090] Of particular interest is the treatment of diseases associated with mitochondrial complex I deficiency / multiple deficiencies. Some diseases are associated solely with dysfunction of complex I, while other diseases are associated with multiple defects, e.g., several mitochondrial complexes.

[0091] As is known in the art, the mitochondrial respiratory chain is the basis of oxidative phosphorylation, a critical cellular process that uses oxygen and simple sugars to generate adenosine triphosphate (ATP), the cell's primary energy source. Five protein complexes, each consisting of several proteins, are involved in this process. The complexes are designated Complex I, Complex II, Complex III, Complex IV, and Complex V. The first enzyme in the respiratory chain, Complex I (CI, or NADH dehydrogenase, NADH-coenzyme Q reductase), is a very large protein complex (approximately 1000 kDa) composed of at least 44 subunits, seven of which are encoded by mitochondrial DNA (ND1-ND6 and ND4L).

[0092] According to certain embodiments, Alverine or one of its derivatives can be used to treat so-called "primary" mitochondrial diseases, i.e., those due to identified genetic abnormalities in at least one subunit of Complex I linked to either pathogenic mitochondrial or nuclear DNA mutation(s). These pathologies are associated with neurological, myocardial, or ophthalmological symptoms and are the tissues or organs most affected in such mitochondrial diseases, even though other organs or tissues may also be affected.

[0093] According to another particular embodiment, Alverine or one of its derivatives can be used to treat so-called "secondary" mitochondrial diseases. In such cases, the genetic abnormality does not directly affect complex I, but the pathology may affect mitochondrial function, particularly reducing the enzymatic activity of this complex I. Such diseases may also be due to non-genetic causes, such as environmental factors or aging. This is particularly true in Parkinson's disease or other age-related neurodegenerative diseases.

[0094] According to one embodiment, the mitochondrial deficiency or dysfunction is due to a genetic disease.

[0095] Genetic diseases, by definition, are diseases that result from one or more genetic defects (or mutations) in one or more genes. Genetic defects can affect mitochondrial DNA and / or nuclear genes. Genetic defects that cause mitochondrial diseases are point mutations, resulting in codon changes. However, diseases can also be associated with the deletion or insertion of one or more bases or codons.

[0096] According to certain embodiments, the disease results from one or more genetic defects (or mutations) in one or more genes involved in complex I functionality.

[0097] A non-limiting list of such genes is as follows: - Complex I structural genes, in particular MTND1 (or ND1), MTND2 (or ND2), MTND3 (or ND3), MTND4 (or ND4), MTND5 (or ND5), MTND6 (or ND6), MTND4L (or ND4L), NDUFA1, NDUFA2, NDUFA3, NDUFA4, NDUFA5, NDUFA6, NDUFA7, NDUFA8, NDUFA9, NDUFA10, NDUFA11, NDUFA12, NDUFA13, NDUFAB1, NDUFB1, NDUFB2, NDUFB3, NDUFB4, NDUFB5, NDUFB6, NDUFB7, NDUFB8, NDUFB9, NDUFB10, NDUFB11, NDUFC1, NDUFC2, NDUFS1, NDUFS2, NDUFS3, NDUFS4, NDUFS5, NDUFS6, NDUFS7, NDUFS8, NDUFV1, NDUFV2, NDUFV3; - Complex I assembly genes, in particular NDUFAF1, NDUFAF2, NDUFAF3, NDUFAF4, NDUFAF5, NDUFAF6, NDUFAF7, NDUFAF8, NUBPL, ACAD9, TMEM126B, FOXRED1, ECSIT, AIF, TIMMDC1 Includes.

[0098] As an example, Alverine or one of its derivatives can be used to treat genetic diseases in which the genetic defect is related to ND3, ND6, NDUFV1 or NDUSF8.

[0099] Exemplary mutations are the NDUFV1 mutations c.1162+2A>C and c.1156C>T resulting in a p.Arg386Cys amino-changing substitution, or the NDUFV1 mutation resulting in a p.Ala341val amino-changing substitution, which primarily cause neuropathy or Leigh syndrome.

[0100] Several mitochondrial diseases of genetic origin, particularly those associated with mitochondrial complex I deficiency, are well documented in the prior art.

[0101] LHON syndrome, or Leber's hereditary optic neuropathy, usually occurs in young adults. Onset is abrupt, with a rapid loss of central vision, corresponding to a decline in central vision. In most cases, peripheral vision persists, almost like a halo of vision surrounding the blind field. The disease is caused by a homoplasmic mutation in the gene encoding the respiratory chain complex I subunit. In fact, the following mitochondrial DNA mutations, m.11778G>A, m.3460G>A, and m.14484T>C, represent approximately 95% of LHON mutations.

[0102] Leigh syndrome (or LS) is a progressive, severe neurodegenerative disorder. Affected individuals typically exhibit global developmental delay or regression, hypotonia, ataxia, dystonia, nystagmus, or ophthalmologic abnormalities such as optic atrophy. Leigh syndrome can also have adverse multi-organ system effects on cardiac, hepatic, gastrointestinal, and renal tissues. Biochemical studies in individuals with Leigh syndrome tend to show elevated lactate and abnormalities in mitochondrial oxidative phosphorylation. Leigh syndrome may be associated with mutations in genes encoding complex I subunits, such as the NDUFV1 mutation or the MTND5 m.13513G>A mutation.

[0103] MELAS syndrome, which includes mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes, is a genetically heterogeneous mitochondrial disorder with a diverse clinical phenotype. The disorder is characterized by central nervous system involvement, including seizures, hemiplegia, hemianopsia, cortical blindness, and transient vomiting. The syndrome is caused by the m.3243A>G mutation in mitochondrial DNA, i.e., tRNA Leu (UUR) MELAS syndrome has been initially linked to the mitochondrial DNA mutation m.3260A>G, which alters the translation of complex I mRNA into protein, thus reducing the amount of structural proteins of complex I, such as ND6. Leu (UUR) The m.3260A>G mutation may also result in other clinical phenotypes, including maternally inherited myopathy and cardiomyopathy.

[0104] Of particular interest is the treatment of genetic disorders that have been shown to be associated with complex I deficiency, including MELAS syndrome, Leigh syndrome, and Leber's hereditary optic neuropathy. Note that such disorders may be associated with other symptoms, such as cardiac, myopathic, or neurological clinical phenotypes.

[0105] More generally, Alverine or one of its derivatives can be used to treat mitochondrial dysfunction, in particular mitochondrial dysfunction associated with complex I deficiency, which is characterized by a loss of efficiency in the electron transport chain and a decrease in the synthesis of high-energy molecules such as adenosine-5'-triphosphate (ATP), is characteristic of aging, and is a hallmark of essentially all chronic diseases.

[0106] These diseases include: - Neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (Lou Gehrig's disease), and Friedreich's ataxia; - cardiovascular disease, e.g., atherosclerosis, and other heart and vascular conditions; - diabetes and metabolic syndrome; - autoimmune diseases, such as multiple sclerosis, systemic lupus erythematosus, and type 1 diabetes; - Neurobehavioral and psychiatric disorders, such as autism spectrum disorders, schizophrenia, bipolar disorder and mood disorders; - Gastrointestinal disorders; - Fatiguing illnesses, such as chronic fatigue syndrome and Gulf War illness; - Musculoskeletal disorders, e.g., fibromyalgia and skeletal muscle hypertrophy / atrophy; - muscular dystrophy; - Cancer; and - Chronic infection.

[0107] According to a specific embodiment, functional gastrointestinal motility disorders are outside the definition of the diseases to be treated in the framework of the present invention.

[0108] According to one embodiment, the composition according to the invention is associated with at least another compound for the treatment of the same disease. The composition according to the invention and said compound can be administered simultaneously or separately over time, taking into account their specific properties, in particular their bioavailability.

[0109] According to a particular embodiment, the present invention relates to a composition, preferably a pharmaceutical composition or a pharmaceutical product, containing the above-mentioned compound and potentially other active molecules (other gene therapy proteins, chemical groups, peptides or proteins, etc.) for the treatment of the same or different diseases, preferably the same diseases.

[0110] Preferably, the pharmaceutical composition and at least one compound according to the present invention for the treatment of the same or different diseases are administered simultaneously, separately or chronologically spread to treat the same or different diseases.

[0111] More generally, with respect to mitochondrial diseases, the additional compound capable of improving mitochondrial function can be administered at the same time or at different times. In the case of simultaneous administration, the two compounds can be associated in the same composition.

[0112] Examples of such additional compounds are natural supplements such as L-carnitine, alpha-lipoic acid (α-lipoic acid [1,2-dithiolane-3-pentanoic acid]), coenzyme Q10 (CoQ10 or ubiquinone), riboflavin (a B2 vitamin) reduced nicotinamide adenine dinucleotide (NADH), L-arginine, and the like, possibly in combination.

[0113] For example, examples of compounds used in the case of MELAS syndrome are nitric oxide (NO) precursors such as L-arginine and citrulline.

[0114] Subjects who may benefit from the compositions of the present invention include all patients who have a disease associated with mitochondrial dysfunction, particularly mitochondrial dysfunction associated with Complex I deficiency, and who are diagnosed with or at risk of developing such a disease.

[0115] Subjects to be treated with the compositions according to the invention can be selected based on various criteria. In relation to mitochondrial dysfunction, particularly complex I deficiency, several tests can be performed, for example: - at the biochemical level: O2 consumption and / or mitochondrial complex I activity (disclosed in Examples 8-9 below) can be measured on the basis of biopsies, in particular muscle or skin biopsies of the subject. The activity of other complexes of the respiratory chain can be further assessed to determine whether mitochondrial dysfunction is due solely to complex I deficiency; - At the genetic level: sequencing DNA extracted from blood, cells or biopsy samples, e.g., skin, makes it possible to identify one or more molecular abnormalities, in particular mutations or deletions / insertions in the preferred genes listed above. Alternatively, the corresponding protein expression or activity can be assessed by any method known to those skilled in the art (e.g., Western blotting).

[0116] The target of the present invention is to provide a safe (non-toxic) treatment. A further objective is to provide an effective treatment that makes it possible to postpone, delay or prevent the onset of the disease and to improve the patient's phenotype, which can be easily monitored at a clinical level, as disclosed below.

[0117] In the subject, the composition according to the present invention comprises: - To improve mitochondrial function, especially mitochondrial respiration; - To improve growth; - To improve muscle function; - To improve vision and / or hearing; - To improve respiratory function; - To improve heart, liver or kidney function; - To improve brain function; - To improve digestion, and / or - It can be used to prolong survival and, more generally, improve quality of life and life expectancy.

[0118] According to one aspect, the present invention relates to a method for improving mitochondrial function, particularly Complex I activity, advantageously without adverse effects, comprising administering to a subject in need thereof a therapeutic amount of the composition disclosed above.

[0119] Advantageously, the improvement is observed for up to 1 month, or 3 months, or 6 months, or 9 months after treatment begins, and more advantageously for up to 1 year, 2 years, 5 years, 10 years, or over the entire lifetime of the subject after treatment begins.

[0120] In one embodiment, the improvement results in a decrease in severity and / or frequency of symptoms and / or a delay in their onset.

[0121] The enhancement can be achieved, for example, in the case of MELAS, by methods known in the art: - Assessment of lactate levels, particularly ventricular lactate, as measured, for example, by magnetic resonance spectroscopy (MRS); - Assessment of quality of life and / or life expectancy by clinical scales, e.g., NMDAS (Newcastle Mitochondrial Disease Scale for Adults) score or SF-36 (Short Form Health Survey) score; - Assessment of brain changes, for example, using magnetic resonance imaging (MRI); - Assessment of changes in muscle activity using physical examinations such as the 6-minute walk test; - Assessment of changes in intravenous lactate and GDF 15 concentrations; - Evaluation of changes in mtDNA heteroplasmy in urine and blood can be evaluated based on the following.

[0122] The appropriate parameters for a particular case can be adapted depending on the disease.

[0123] The claimed treatment therefore makes it possible to improve the clinical condition and the various parameters disclosed above compared to untreated subjects.

[0124] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology that are well within the skill of those in the art. Such techniques are fully explained in references such as "Molecular Cloning: A Laboratory Manual," 4th Edition (Sambrook, 2012); "Oligonucleotide Synthesis" (Gait, 1984); "Culture of Animal Cells" (Freshney, 2010); "Methods in Enzymology" and "Handbook of Experimental Immunology" (Weir, 1997); "Gene Transfer Vectors for Mammalian Cells" (Miller and Calos, 1987); "Short Protocols in Molecular Biology" (Ausubel, 2002); "Polymerase Chain Reaction / Principles, Applications and Troubleshooting" (Babar, 2011); and "Current Protocols in Immunology" (Coligan, 2002). These techniques are applicable to the production of the polynucleotides and polypeptides of the invention and, as such, may be considered in making and practicing the invention. The following sections describe particularly useful techniques for particular embodiments.

[0125] Without further description, it is believed that one of ordinary skill in the art can, using the foregoing description and the following illustrative examples, make and utilize the present compounds and practice the claimed methods. [Example]

[0126] The present invention and its advantages will be better understood from the examples presented below, which are supported by the accompanying drawings. In particular, the present invention is illustrated with respect to the effects of alverine citrate (ALV) in various model organisms for mitochondrial diseases, as well as in patient cell lines.

[0127] ALV has been shown to be active against several complex I subunits from different organisms carrying mutations in the following subunits: NDUFV1 (Schuelke et al., 1999), NDUFS8 (Procaccio et al., 2004), ND2 or ND3 (Sarzi et al., 2007), and ND6 (John et al., 1992) subunits.

[0128] In addition, 4-hydroxyalverine (4-hydroxyALV), a metabolite of ALV, was shown to be active against Podospora anserina nuo-51 gene carrying the mutation A357V.

[0129] Furthermore, the efficacy of Alverine has been demonstrated in other mitochondrial yeast mutants, however, these examples are by no means limiting. [Brief explanation of the drawings]

[0130] [Figure 1] FIG. 1 shows the effect on growth rate of the heat-sensitive mutation A357V in the Podospora anserina nuo-51 gene, which mimics the pathogenic NDUFV1 A341V mutant in humans. [Figure 2]This figure shows the effect of the thermosensitive mutation A357V in the Podospora anserina nuo-51 gene, which mimics the human pathogenic NDUFV1 A341V mutant, on respiration (O2) and complex I activity (CI) at 31.5 °C. CS: citrate synthase activity. Asterisks (** or ***) indicate statistically significant differences compared to the wild-type strain (WT). [Figure 3] This figure shows the effects of ALV (A) and 4-hydroxyALV (B) compared with DMSO (C) on the thermosensitive growth at 33°C of the nuo-51A357V mutant of Podospora anserina, which mimics the human pathogenic mutation NDUFV1A341V. [Figure 4] Figure 1 shows the effect of various concentrations of ALV (0.001-10 μM; VEH=0) on the growth rate of nuo-51A357V (NDUFV1) at 31.5° C., Δnuo-19 (NDUFS7) and nd2-nd3 (ND) mutant strains at 28° C. Asterisks (* or **) indicate statistically significant differences relative to untreated cultures (VEH=vehicle). [Figure 5] Figure 1 shows the effect of ALV 1 μM on the O2 consumption rate of nuo-51A357V (NDUFV1) at 31.5° C. Asterisks (**) indicate statistically significant differences relative to untreated cultures (VEH = vehicle). [Figure 6] Figure 1 shows the effect of 1 μM ALV on complex I activity (CI) of nuo-51A357V (NDUFV1) at 31.5°C. Asterisks (***) indicate statistically significant differences relative to untreated cultures (VEH = vehicle). CS: citrate synthase activity. [Figure 7] Figure 1 shows the effect of 1 μM ALV on C. elegans nuo-1A352V(NDUFV1) mutant progeny. Asterisks (**) indicate statistically significant differences relative to untreated cultures. [Figure 8] Figure 1 shows the effect of 1 μM ALV on the progeny of wild-type worms exposed to NDUFV1 or NDUFS8 RNAi. Asterisks (**) indicate statistically significant differences relative to untreated cultures. [Figure 9]Figure 1 shows the determination of the maximum ALV concentration that is non-toxic to the growth of NDUFV1 mutant cells carrying the compound heterozygous mutations c.1162+2A>C and c.1156C>T (p.Arg386Cys). ALV concentration range of 30 nM to 30 μM. [Figure 10] Figure 1 shows the determination of active ALV concentration in citrate synthase activity of NDUFV1 mutant cells harboring the compound heterozygous mutations c.1162+2A>C and c.1156C>T (p.Arg386Cys). An asterisk (*) indicates a statistically significant difference compared to untreated cells (vehicle). [Figure 11] Figure 1 shows the determination of active ALV concentration in complex I enzyme activity of NDUFV1 mutant cells harboring the compound heterozygous mutations c.1162+2A>C and c.1156C>T (p.Arg386Cys). Asterisks (* or ** or ***) indicate statistically significant differences compared to untreated cells (vehicle). [Figure 12] Figure 1 shows the determination of active ALV concentration in complex I enzyme activity (CI) normalized with respect to citrate synthase activity (CS) in NDUFV1 mutant cells harboring the compound heterozygous mutations c.1162+2A>C and c.1156C>T (p.Arg386Cys). Asterisks (* or **) indicate statistically significant differences relative to untreated cells (vehicle). [Figure 13] Figure 1 shows the determination of the maximum ALV concentration that is non-toxic to the growth of ND6 mutant cells harboring the m.14484T>C (p.Met64Val) mitochondrial DNA mutation. ALV concentrations range from 30 nM to 10 μM. Veh: vehicle. n=4 + / - SEM. *P<0.05. [Figure 14] Measurement of complex I enzyme activity of ND6 mutants in the presence of Alv. (A): Normalized to citrate synthase activity of ND6 cell line after 48 h treatment with different concentrations of Alv. (B): Enzymatic measurement of citrate synthase activity of ND6 mutant cell line. n=4+ / -SEM. *P<0.05. [Figure 15]Figure 1 shows the determination of lactate production in control fibroblasts, NDUFV1 and ND6 mutant fibroblast cell lines after 48 hours of exposure to Alv. (A) Control cells; NDUFV1 (B) and ND6 (C). (A) and (B): n=3. (C): n=4 + / - SEM. *P<0.05. [Figure 16] FIG. 1 shows the effect of alverine on the growth of different mutant yeast strains on solid respiratory medium as detected by the halo test. [Figure 17] FIG. 1 shows the determination of the active range of ALV concentrations that result in suppression of the respiratory growth defect of taz1 mutant yeast strains.

[0131] Examples 1 to 5 Podospora anserina model The filamentous fungus Podospora anserina strain used in Examples 1-5 contains a specific mutation modeling a human mutation in the NDUFV1 subunit of complex I that results in mitochondrial disease. This strain exhibits a growth defect at non-permissive temperatures above 31.5 °C.

[0132] Podospora anserina mutant strain: 〇nuo-51 A357V → Nourseothricin resistance cassette (Nat R ) related to the human NDUFV1 strain, the A357V mutation was introduced into the nuo-51 gene of the wild-type strain S. A341V To fully mimic the disease, the NDI-1 and AOX genes were inactivated (El-Khoury et al., 2008). Strain genotype: S, mat-, nuo-51 A357V , Δndi-1, Δaox, Nat R , Hygro R The nd2-nd3 → Wa32-LL strain harbors a mitochondrial plasmid integrated into the 5′-UTR of the mitochondrial cotranscription unit nd2 / nd3, which downregulates the levels of the corresponding subunits ND2 and ND3 ( Maas et al., 2007 ). o Δnuo-19 → The gene encoding the homologue of the human PSST (NDUFS7) subunit was inactivated in the S wild-type strain ( Maas et al., 2010 ).

[0133] Example 1 Human pathogenic mutations in NDUFV1 A341V Effects of the thermosensitive mutation A357V in the nuo-51 gene of Podospora anserina mimicking the growth rate (1), respiration, and complex I activity (2) Materials and Methods Mycelia are grown on Petri dishes containing solid synthetic M2 medium at 28° C., starting from ascospores. Cultures are grown on solid synthetic medium (M2, http: / / podospora.i2bc.paris-saclay.fr / ) and growth rate is measured in centimeters of mycelial growth per day. In Figure 1, growth rate is expressed relative to the wild type.

[0134] For oxygen consumption, a 40 mg sample of mycelium grown at 31.5 °C is introduced into an Oxytherm device (Hansatech electrode). After 2 minutes of registration, the sample is withdrawn and the protein content is estimated by Bradford protein assay (Biorad). Respiration is registered as nmol oxygen consumed / min / μg protein and then expressed relative to the wild type. At least three samples of each strain were examined per experiment. Experiments were repeated at least five times.

[0135] Complex I activity (CI) was performed on crude mitochondria obtained after mixing mycelium with glass beads in 0.4 M sucrose buffer and differential centrifugation. NADH consumption rates in the presence and absence of rotenone were recorded spectrophotometrically (340-380 nm) using 25 µg of mitochondria starting with NADH and quinone addition. Citrate synthase activity (CS) was determined in an equivalent sample of mitochondria to express Complex I activity as the ratio of NADH dehydrogenase-rotenone-sensitive activity to citrate synthase activity (CI / CS).

[0136] result The results are shown in Figures 1 and 2. nuo-51 A357V The strain is thermosensitive compared to the wild-type strain, with a 30% reduction in growth rate at 31.5°C and no growth at 33°C (Fig. 1). nuo-51 A357V The mutant strain exhibits a 40% reduction in oxygen consumption and a 70% reduction in complex I activity (Fig. 2 ).

[0137] Example 2 NDUFV1 A341V Podospora anserina nuo-51 mimics human pathogenic mutations in A357V Effect of ALV and 4-hydroxy ALV on the heat-sensitive growth of mutants Materials and Methods The molecules (ALV, 4-hydroxyALV, and DMSO) were tested in a drug drop test to determine the activity of nuo-51 A357V Mutants were tested for their ability to restore the heat-sensitive growth defect. Mutant mycelia scraped from 2-day cultures on M2 plates at 28°C were mixed in a FastPrep apparatus and spread onto fresh square plates. A 6 mm cellulose disk was then dropped and spotted with various compounds at 10 mM in DMSO, and the plates were incubated at 33°C. Within 4-6 days, a presumptive resumption of growth (i.e., a halo) around the filter was observed.

[0138] result The results are shown in Figure 3. Alverine citrate (A: upper left corner) and 4-hydroxyalverine, the first metabolite in the degradation pathway of alverine (B: upper right corner), were metabolized by nuo-51 without showing any toxicity. A357V The fact that the same effect was observed with both compounds indicates that the compound, alverine citrate, is the active alverine.

[0139] Example 3 nuo-51 A357V Effect of ALV on the growth rate of (NDUFV1), Δnuo-19 (NDUFS7), and nd2-nd3 (ND) mutant strains Materials and Methods The mutant strain was the thermosensitive nuo-51 strain grown in the presence of various concentrations of ALV at 31.5°C (as determined in Example 1) for 3 to 6 days. A357V Except for the strains, they were grown at 28° C. Growth rates were calculated as described in Example 1.

[0140] Experiments were performed at least in triplicate. Differences between treated versus untreated cultures (VEH) were assessed according to standard deviations (error bars).

[0141] result The data are shown in Figure 4. nuo-51 A357V The nd2-nd3 and nd3 mutant strains show increased growth rates in the presence of ALV. Conversely, ALV does not increase the growth rate of the Δnuo-19 strain (complete absence of functional complex I). The optimal ALV dose-response determined from five independent experiments was nuo-51. A357V 1 μM for the strain and used for further experiments. Toxicity cannot be detected below 10 μM.

[0142] Example 4 nuo-51 A357V Effect of ALV on O2 consumption rate of (NDUFV1) Materials and Methods nuo-51 A357V The mutant strains were grown at 31.5°C in the absence (VEH) or presence of 1 μM ALV (optimal concentration determined in Example 3), and 40 mg mycelium was introduced into an Oxytherm apparatus (Hansatech electrode) as described in Example 1.

[0143] Experiments were performed at least five times and error bars represent standard deviation. Differences between treated vs. untreated cultures (VEH) were assessed using Pearson's chi-square test, with a significant p-value of <0.05.

[0144] result The results are shown in Figure 5. nuo-51 A357V The respiration rate of the mutant is significantly increased in the presence of 1 μM ALV.

[0145] Example 5 nuo-51 A357V Effect of ALV on Complex I activity of (NDUFV1) Materials and Methods nuo-51 A357V The mutant strains were grown at 31.5°C in the presence or absence (VEH) of ALV (1 μM), and rotenone-sensitive NADH dehydrogenase activity was determined in crude mitochondrial extracts as described in Example 1.

[0146] Experiments were performed at least five times, and error bars represent standard deviations. Differences between treated versus untreated cultures (VEH) were assessed using Pearson's chi-square test, with a significant p-value of <0.05.

[0147] result The results are shown in Figure 6. nuo-51 A357V The complex I activity of the mutant is significantly increased in the presence of 1 μM ALV.

[0148] Examples 6 and 7 Caenorhabditis elegans model The C. elegans nematodes used in Examples 6 and 7 are the wild-type BRISTOL N2 strain from the Caenorhabditis Genetics Center (CGC) Consortium. A341V Imitating nuo-1 A352V The mutant strain LB25 carrying the mutation (Δnuo-1, ex:nuo-1(A352V)) was constructed by B. Lemire (Canada; Grad and Lemire, 2004 ).

[0149] Example 6 C. elegans nuo-1 A352V (NDUFV1) Effects of ALV in offspring Materials and Methods Individual L4 animals (F0) developed from the L1 stage with or without 1 μM ALV were transferred to separate wells in a microplate, monitored daily during egg laying, and transferred to fresh wells to separate them from their offspring. Two to three days after hatching, adult offspring (F1-adult) were counted.

[0150] More than 30 F0-adult offspring were recorded from three independent experiments, and error bars represent standard deviations. Differences between treated versus untreated cells were assessed using Student's t-test, with significant p values ​​<0.05.

[0151] result The results are shown in Figure 7. In the presence of 1 μM ALV in the medium, nuo-1 A352V There is a significant increase in the number of adult worm progeny, i.e. F1 adults obtained by F0 adults (p=0.02).

[0152] Example 7 Effects of ALV in the progeny of wild-type worms exposed to NDUFV1 or NDUFS8 RNAi Materials and Methods Downregulation of NDUFV1 or NDUFS8 in wild-type worms (N2) was achieved by RNAi via feeding (Kamath RS and Ahringer J., 2003). Wild-type worms were synchronously developed from L1 to L4 stages under RNAi conditions with or without 1 μM ALV. Individual L4 animals were then transferred to separate wells in microplates (same medium and RNAi conditions) and monitored within the next 3–7 days.

[0153] RNAi of NDUFV1 (C09H10.3) and NDUFS8 (T20H4.5) resulted in few F0 adults capable of producing larval offspring. Therefore, the number of F0 adults capable of producing larval offspring was monitored. More than 30 F0 adult offspring were recorded in three independent experiments (t-test, p = 0.02). The efficiency of RNAi was determined by real-time qRT-PCR (20% residual mRNA of NDUFV1 or NDUFS8 in F0 adults under RNAi conditions).

[0154] result The results are shown in Figure 8. Under RNAi conditions that reduce the expression of NDUFV1 (part of the N module) or NDUFS8 (part of the Q module), 1 μM ALV can increase the progeny.

[0155] (Examples 8 to 12) Human cell model Since Alverine (ALV) was found to have a positive effect based on P. anserina mutant strains and helminth mutants, it was next tested in human mutant cells derived from patients: skin fibroblasts carrying a complex I mutation (NDUFV1 mutation of c.1162+2A>C, c.1156C>T; (p.Arg386Cys)) were used in Examples 8 and 9.

[0156] Example 8 Effect of ALV (concentrations from 30nM to 30.MU.M) on cell proliferation of NDUFV1 fibroblast mutant cells carrying the compound heterozygous mutations c.1162+2A>C and c.1156C>T (p.Arg386Cys) Materials and Methods NDUFVI mutant fibroblasts were cultured in 24-well plates in low-glucose medium (0.5 g / L) DMEM-F12 supplemented with 10% fetal bovine serum and 1% glutamine, as described by Leman et al. (2015). Growth and confluence were monitored in real time in wells seeded at the same cell density in an automated manner at 37°C in the presence of 5% CO2 for 96 hours. Mutant cells were treated with different concentrations of ALV (30 nM to 30 μM) relative to vehicle. A growth time course revealed a concentration-dependent treatment effect according to cell confluence.

[0157] result The results are shown in Figure 9. Cell proliferation of NDUFV1 mutant fibroblasts harboring the compound heterozygous mutations c.1162+2A>C and c.1156C>T (p.Arg386Cys) exposed to different ALV drug concentrations (30 nM to 30 μM) was reduced at 10 μM and 30 μM concentrations of ALV compared to vehicle.

[0158] Example 9 Determination of active ALV concentration on complex I enzyme activity on NDUFV1 fibroblast mutant cells carrying the compound heterozygous mutations c1162+2A>C and c1156C>T (p.Arg386Cys) Materials and Methods Complex I mutant cells carrying compound heterozygous NDUFV1 mutations causing complex I deficiency were cultured in standard DMEM high-glucose medium (4.5 g / L) or low-glucose medium (0.5 g / L) supplemented with 10% fetal bovine serum, 1% glutamine, and 50 μg / ml uridine at 37°C in the presence of 5% CO2, as described elsewhere (Leman et al., 2015). To optimize the effect of drug concentrations, cells were shifted to low-glucose medium (0.5 g / L) supplemented with various concentrations of alverine (10 nM to 10 μM) or vehicle (DMSO) for 48 hours (to force cells to rely on OXPHOS rather than glycolysis).

[0159] Complex I enzyme activity was measured at 37°C on a UVmc2 spectrophotometer (SAFAS) as described (Leman et al., 2015; Desquiret-Dumas et al., 2012). For complex I enzyme activity, 500,000 cells were sonicated (6 cycles of 5 seconds) and then incubated at 37°C in reaction medium (100 mM KH2PO4, pH 7.4, 1 mM KCN, 2 mM NaN3, 1 mg / ml BSA, 0.1 mM ubiquinone-1, and 0.075 mM DCPIP). The reaction was initiated by the addition of 0.15 mM NADH, and the rate of disappearance of DCPIP was measured at 600 nm for 2 minutes. Nonspecific activity was determined in the presence of rotenone (5 μM).

[0160] Complex I enzyme activity was normalized with respect to citrate synthase (CS) activity, which is considered a marker of mitochondrial mass. Citrate synthase activity was measured by adding 100,000 cells to a prewarmed reaction mixture consisting of 0.15 mM 5,5'-dithiobis-2-nitrobenzoic acid (DTNB), 0.5 mM oxaloacetate, 0.3 mM acetyl-coA, and 0.1% Triton X100, and the rate of appearance of CoA-SH at 412 nm was assessed after DTNB reduction.

[0161] Experiments were performed at least in triplicate, and error bars represent standard deviation. Differences between treated versus untreated (DMSO) cells were assessed using Student's t-test, with a significant p-value of <0.05.

[0162] result The results are shown in Figures 10 to 12. Asterisks (*, **, or ***) indicate statistically significant differences compared to untreated cells (vehicle).

[0163] The data show that ALV has a beneficial effect on complex I enzyme activity in NDUFV1 mutant cells at concentrations of 100 nM and 300 nM, and ALV concentrations from 300 nM up to 10 μM do not have a detrimental effect on complex I activity in mutant cells.

[0164] Example 10 The effect of ALV (concentrations from 30nM to 10μM) on cell proliferation of ND6 fibroblast mutant cell lines carrying m.14484T>C (p.Met64Val) in the MT-ND6 gene Complex I is encoded by both the nuclear and mitochondrial genomes. Having shown that Alv has a positive effect in fibroblasts harboring mutations in the nuclear-encoded subunit, NDUFV1, we next tested it in fibroblast complex I-deficient cells harboring the homoplasmic m.14484T>C (p.Met64Val) mitochondrial DNA mutation, which has been shown to cause LHON disease.

[0165] Materials and Methods ND6 mutant fibroblasts were cultured in 24-well plates in low-glucose medium (0.5 g / L) DMEM-F12 supplemented with 10% fetal bovine serum and 1% glutamine, as described by Leman et al. (2015). Real-time growth and confluence were monitored in an automated manner using a CCD camera on an IncuCyte® live-cell analysis system device in wells seeded at the same cell density for 96 hours at 37°C in the presence of 5% CO2. Mutant cells were treated with different concentrations of ALV (30 nM to 10 μM) versus vehicle. A growth time course revealed a concentration-dependent effect of treatment according to cell confluence.

[0166] result The results are shown in Figure 13. Cell proliferation of ND6 mutant fibroblasts harboring the homoplasmic m.14484T>C (p.Met64Val) mitochondrial DNA mutation exposed to different ALV drug concentrations (30 nM to 10 μM) was unchanged at concentrations of 30, 100, and 300 nM. However, a significant increase in doubling time (i.e., a decrease in cell proliferation) was observed at ALV concentrations of 1 μM or higher compared to vehicle.

[0167] Example 11 Measurement of complex I enzyme activity for ND6 mutant cell lines harboring the homoplasmic m.14484T>C (p.Met64Val) mitochondrial DNA mutation after Alv treatment Materials and Methods Complex I mutant cells carrying the homoplasmic m.14484T>C (p.Met64Val) mitochondrial DNA mutation, which causes Complex I deficiency, were cultured in standard DMEM high-glucose medium (4.5 g / L) or low-glucose medium (0.5 g / L) supplemented with 10% fetal bovine serum, 1% glutamine, and 50 μg / ml uridine at 37°C in the presence of 5% CO2, as described elsewhere (Leman et al., 2015). To optimize the effect of drug concentrations, cells were shifted to low-glucose medium (0.5 g / L) supplemented with various concentrations of alverine (30 nM to 10 μM) or vehicle (DMSO) for 48 hours (to force cells to rely on OXPHOS rather than glycolysis).

[0168] Complex I enzyme activity was measured at 37°C on a UVmc2 spectrophotometer (SAFAS) as described (Leman et al., 2015; Desquiret-Dumas et al., 2012). For complex I enzyme activity, 500,000 cells were sonicated (6 cycles of 5 seconds) and then incubated at 37°C in reaction medium (100 mM KH2PO4, pH 7.4, 1 mM KCN, 2 mM NaN3, 1 mg / ml BSA, 0.1 mM ubiquinone-1, and 0.075 mM DCPIP). The reaction was initiated by the addition of 0.15 mM NADH, and the rate of disappearance of DCPIP was measured at 600 nm for 2 minutes. Nonspecific activity was determined in the presence of rotenone (5 μM).

[0169] Complex I enzyme activity was normalized with respect to citrate synthase (CS) activity, which is considered a marker of mitochondrial mass. Citrate synthase activity was measured by adding 100,000 cells to a prewarmed reaction mixture consisting of 0.15 mM 5,5'-dithiobis-2-nitrobenzoic acid (DTNB), 0.5 mM oxaloacetate, 0.3 mM acetyl-coA, and 0.1% Triton X100, and the rate of appearance of CoA-SH at 412 nm was assessed after DTNB reduction.

[0170] Experiments were performed at least in triplicate, and error bars represent standard deviation. Differences between treated versus untreated (DMSO) cells were assessed using Student's t-test, with a significant p-value of <0.05.

[0171] result The results are shown in Figure 14. An asterisk (*) indicates a statistically significant difference relative to untreated cells (vehicle).

[0172] The data reveal that ALV at concentrations of 30 nM and 300 nM has a beneficial effect on complex I enzyme activity in ND6 mutant cells normalized to citrate synthase activity.

[0173] Example 12 Determination of lactate production in NDUFV1 and ND6 mutant cell lines exposed to different concentrations of Alv from 30nM to 10.MU.M The mutant cell line has a preferential anaerobic glycolytic metabolism compared to control cells. Lactate production was assessed after 48 h of exposure in the presence of different concentrations of Alv, ranging from 30 nM to 10 µM.

[0174] Materials and Methods A total of 15,000 cells / well were seeded into 96-well plates and grown at 37°C in the presence of 5% CO2. After 24 h, cells were treated with different concentrations of Alv, ranging from 30 nM to 10 μM. After 48 h, supernatants were collected and assayed for lactate using a lactate determination kit (Abcam kit ab65330) according to the manufacturer. Lactate concentrations were measured using a microplate spectrophotometer (CLARIOstar instrument, BMG Labtech).

[0175] result The results are shown in Figure 15. Alv did not alter lactate production in the control fibroblast cell line (Fig. 15A) and the NDUFV1 cell line (Fig. 15B). Lactate production in the ND6 cell line (Fig. 15C) was significantly increased at the 1 μM and 3 μM concentrations compared to vehicle.

[0176] (Examples 13 to 14) Efficacy of Alverine in other mitochondrial yeast mutants Example 13 Effect of alverine on the growth of various mutant yeast strains. Materials and Methods Yeast mutant strains: A taz1Δ yeast strain was constructed by replacing the TAZ1 open reading frame with that of TRP1 in the W303-1A strain (MATa ade 2-1 ura3-1 his311,15 trp1-1 leu2-3,112 can1-100) (de Taffin de Tilques, M. et al., 2018). A sym1Δ yeast strain was constructed by replacing the SYM1 open reading frame with that of kanMX6 in the W303-1A strain (MATa ade 2-1 ura3-1 his311,15 trp1-1 leu2-3,112 can1-100). fmc1 MC6 MATa ade2-1 his3-11,15 trp1-1 leu2-3,112 ura3-1 fmc1::HIS3 [Δi ER OR] (Schwimmer, C. et al., 2005). NARP MR14 MATa ade2-1 his3-11,15 trp1-1 leu2-3,112 ura3-1 CAN1 arg8::HIS3 ρ+ atp6-L183R (Rak, M. et al., 2007). The shy1Δ yeast strain was constructed by replacing the SHY1 open reading frame with that of HIS in the W303-1A strain (MATa ade 2-1 ura3-1 his311,15 trp1-1 leu2-3,112 can1-100) (Barrientos, A. et al., 2002).

[0177] 0.5 OD in liquid YPD rich fermentation medium (1% yeast extract, 0.5% bactopeptone, 2% glucose) 600 200 µL of various yeast mutant strains grown in PBS were spread onto an agar-based solid respiratory medium: either YPG (1% yeast extract, 0.5% bactopeptone, 2% glycerol) for fmc1, shy1, and NARP mutants, or YPE (1% yeast extract, 0.5% bactopeptone, 2% ethanol) for taz1 and sym1 mutants. Next, a small sterile filter was placed on the agar surface, and 100 nmol of alverine (ALV) was added to the filter. The plates were then incubated at 28 °C for shy1 mutants or at 36 °C for fmc1, taz1, sym1, and NARP mutants for 4-5 days and then scanned. DMSO, the complex vehicle, was used as a negative control.

[0178] result As shown in Figure 16, a halo of enhanced growth was observed around the ALV spotted filters, indicating that ALV is effective in all tested yeast models of mitochondrial disease.

[0179] Example 14 Determination of the active range of ALV concentrations that results in suppression of the respiratory growth defect of taz1 mutant yeast strains Materials and Methods Exponentially growing cells were inoculated into fresh, non-fermentable YPG medium, supplemented or not, with increasing concentrations of ALV. These assays were performed in 96-well plates in a BioScreen device. Cell density was tracked over time to determine both the optimal concentration of ALV for its ability to suppress the respiratory growth defect and the concentration at which ALV became toxic. taz1 cells were grown for 70 hours in liquid medium containing 2% glycerol as a carbon source and increasing concentrations of ALV (100 pM to 100 μM).

[0180] result As shown in Figure 17, increased respiratory growth was observed from 100 pM to 10 μM, indicating a very large therapeutic window, with toxicity observed from 100 μM.

[0181] (References) Barrientos A, Korr D, Tzagoloff A. Shy1p is necessary for full expression of mitochondrial COX1 in the yeast model of Leigh's syndrome. EMBO J. 2002 Jan 15;21(1-2):43-52. Barth PG, Van den Bogert C, Bolhuis PA, Scholte HR, van Gennip AH, Schutgens RB, Ketel AG. Desquiret-Dumas V, Gueguen N, Barth M, Chevrollier A, Hancock S, Wallace DC, Amati-Bonneau P, Henrion D, Bonneau D, Reynier P, Procaccio V. Metabolically induced heteroplasmy shifting and L-arginine treatment reduce the energetic defect in a neuronal-like model of MELAS (2012) Biochimica et Biophysica Acta Molecular Basis of Disease. 1822(6):1019-2 De Taffin de Tilques M, Lasserre JP, Godard F, Sardin E, Bouhier M, Le Guedard M, Kucharczyk R, Petit PX, Testet E, di Rago JP, Tribouillard-Tanvier D. Decreasing cytosolic translation is beneficial to yeast and human Tafazzin-deficient cells. Microb Cell. 2018 Feb 18;5(5):220-232. El-Khoury R, ​​Sellem CH, Coppin E, Boivin A, Maas MF, Debuchy R, Sainsard-Chanet A. Gene deletion and allelic replacement in the filamentous fungus Podospora anserina. ( 2008 ) Curr Genet . 53:249-5 Gorman GS, Schaefer AM, Ng Y, Gomez N, Blakely EL, Alston CL, Feeney C, Horvath R, Yu-Wai-Man P, Chinnery PF, Taylor RW, Turnbull DM, McFarland R. Prevalence of nuclear and mitochondrial DNA mutations related to adult mitochondrial disease. Ann Neurol. 2015 May; 77(5):753-9. Grad LI and Lemire BD. (2004) Mitochondrial complex I mutations in Caenorhabditis elegans produce cytochrome c oxidase deficiency, oxidative stress and vitamin-responsive lactic acidosis. Hum. Mol. Genet. 13(3):303-14. Johns DR, Neufeld MJ, Park RD. An ND-6 mitochondrial DNA mutation associated with Leber hereditary optic neuropathy. Biochem Biophys Res Commun. 1992 Sep 30;187(3):1551-7. Kamath RS and Ahringer J. (2003) Genome-wide RNAi screening in Caenorhabditis elegans. Methods 30(4) :313-21. Leman G, Gueguen N, Desquiret-Dumas V, Kane MS, Wettervald C, Chupin S, Chevrollier A, Lebre AS, Bonnefont JP, Barth M, Amati-Bonneau P, Verny C, Henrion D, Bonneau D, Reynier P, Procaccio V. Assembly defects induce oxidative stress in inherited mitochondrial complex I deficiency. Int J Biochem Cell Biol. 2015 May 27; 65:91-103. Maas MF, Sellem CH, Hoekstra RF, Debets AJ, Sainsard-Chanet A. (2007) Integration of a pAL2-1 homologous mitochondrial plasmid associated with life span extension in Podospora anserina. Fungal Genet Biol.; 44(7):659-71. Maas MF, Sellem CH, Krause F, Dencher NA, Sainsard-Chanet A. (2010) Molecular gene therapy: overexpression of the alternative NADH dehydrogenase NDI1 restores overall physiology in a fungal model of respiratory complex I deficiency. J Mol Biol.; 399(1):31-40. Procaccio V, Wallace D.C. Late-onset Leigh syndrome in a patient with mitochondrial complex I NDUFS8 mutations. Neurology (2004) 62:1899. Rak M, Tetaud E, Duvezin-Caubet S, Ezkurdia N, Bietenhader M, Rytka J, di Rago JP. A yeast model of the neurogenic ataxia retinitis pigmentosa (NARP) T8993G mutation in the mitochondrial ATP synthase-6 gene. J Biol Chem. 2007 Nov 23;282(47):34039-47. Sarzi E, Brown M, Lebon S, Chretien D, Munnich A, Rotig A, Procaccio V. A novel recurrent mtDNA mutation in ND3 gene causing Leigh syndrome and dystonia. American Journal of Medical Genetics (2007) 143: 33-41. Schon EA, Santra S, Pallotti F, Girvin ME. Pathogenesis of primary defects in mitochondrial ATP synthesis. Semin Cell Dev Biol. 2001 Dec;12(6):441-8. Schuelke M, et al. Mutant NDUFV1 subunit of mitochondrial complex I causes leukodystrophy and myoclonic epilepsy. Night Genet. 1999; 21(3):260-61. Schwimmer C, Lefebvre-Legendre L, Rak M, Devin A, Slonimski PP, di Rago JP, Rigoulet M. Increasing mitochondrial substrate-level phosphorylation can rescue respiratory growth of an ATP synthase-deficient yeast. J Biol Chem. 2005 Sep 2;280(35):30751-9. Spinazzola A, Viscomi C, Fernandez-Vizarra E, Carrara F, D'Adamo P, Calvo S, Marsano RM, Donnini C, Weiher H, Strisciuglio P, Parini R, Sarzi E, Chan A, DiMauro S, Rotig A, Gasparini P, Ferrero I, Mootha VK, Tiranti V, Zeviani M. MPV17 encodes an inner mitochondrial membrane protein and is mutated in infantile hepatic mitochondrial DNA depletion. Night Genet. 2006 May;38(5):570-5. Wang, K., Takahashi, Y., Gao, Z.-L., Wang, G.-X., Chen, X.-W., Goto, J., Lou, J.-N., Tsuji, S. Mitochondrial ND3 as the novel causative gene for Leber hereditary optic neuropathy and dystonia. Neurogenetics. 10: 337-345, 2009.

Claims

1. A pharmaceutical composition for use in the treatment of mitochondrial diseases of genetic origin, comprising alberin or one of its derivatives, The pharmaceutical composition includes derivatives of alberine, alberine citrate, alberine-d5 citrate, 4-hydroxyalberine (HCl), 4-hydroxyalberine-d5, and 4-hydroxyalberine glucronide.

2. The composition according to claim 1, wherein the composition comprises alberine citrate or 4-hydroxyalberine.

3. The composition according to claim 1 or 2, wherein the disease is a mitochondrial respiratory chain disorder.

4. The composition according to claim 3, wherein the disease is mitochondrial complex I deficiency.

5. The genetic disorder affects the following genes: MTND1 (or ND1), MTND2 (or ND2), MTND3 (or ND3), MTND4 (or ND4), MTND5 (or ND5), MTND6 (or ND6), MTND4L (or ND4L), NDUFA1, NDUFA2, NDUFA3, NDUFA4, NDUFA5, NDUFA6, NDUFA7, NDUFA8, NDUFA9, NDUFA10, NDUFA11, NDUFA12, NDUFA13, NDUFAB1, NDUFB1, NDUFB2, NDUFB3, NDUFB4, NDUFB5, NDUFB6, NDUFB7, NDUFB8, NDUFB9, NDUFB10, NDUFB11, NDUFC1, NDUFC2, NDUFS1, NDUFS2, NDUFS3, NDUFS4, NDUFS5, NDUFS6, The composition according to any one of claims 1 to 4, comprising at least one gene defect in at least one of NDUFS7, NDUFS8, NDUFV1, NDUFV2, NDUFV3, NDUFAF1, NDUFAF2, NDUFAF3, NDUFAF4, NDUFAF5, NDUFAF6, NDUFAF7, NDUFAF8, NUBPL, ACAD9, TMEM126B, FOXRED1, ECSIT, AIF, TIMMDC1, MTTL1, ATP6, TAZ, SURF1, POLG, MPV17, OPA1, COA6, and BCS1L.

6. The composition according to claim 5, wherein the genetic disorder comprises at least one gene defect in ND3, ND6, NDUFV1, NDUFS8, ATP6, TAZ, SURF1, or MPV17.

7. The composition according to any one of claims 1 to 6, wherein the disease is selected from the group consisting of MELAS syndrome, maternal myopathy and cardiac myopathy, NARP syndrome, Ree syndrome, Barth syndrome, mitochondrial DNA depletion syndrome, mitochondrial DNA depletion syndrome 4A (Alpers type), mitochondrial DNA depletion syndrome 4B (MNGIE type), mitochondrial recessive ataxia syndrome, sensory ataxic neuropathy, dysarthria and ophthalmoplegia, spinocerebellar ataxia with epilepsy, progressive extraocular palsy, mitochondrial DNA depletion syndrome-6, Navajo neuropathy, Baer syndrome, mitochondrial DNA depletion syndrome 14, infant cardiac-cerebral myopathy due to cytochrome c oxidase deficiency (COA6 mutation), mitochondrial complex III deficiency nucleus type 1, GRACILE syndrome, Leber's hereditary optic neuropathy, and Bjornstad syndrome.

8. The composition according to claim 7, wherein the disease is selected from the group consisting of NARP syndrome, Barth syndrome, mitochondrial DNA depletion syndrome, Leigh syndrome, Leber's hereditary optic neuropathy, and MELAS syndrome.

9. The composition according to any one of claims 1 to 8, wherein the composition comprises another compound for treating the same disease.

10. The composition according to any one of claims 1 to 9, wherein the composition is administered orally.

11. The composition according to any one of claims 1 to 10, wherein the composition is administered daily.

12. The composition according to any one of claims 1 to 11, wherein the composition is in solid form.

13. The composition according to claim 12, wherein the composition is in the form of a tablet.

14. The composition according to any one of claims 1 to 13, wherein the composition comprises 60 mg of alberine or one of its derivatives.

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