Use of ebselen or one of the derivatives thereof to treat mitochondrial pathologies or dysfunctions

Abstract

The invention relates to the use of ebselen or one of the derivatives thereof to treat mitochondrial pathologies or dysfunctions, in particular associated with deficiencies in complex I.

Description

[0001] USE OF EBSELEN OR ONE OF ITS DERIVATIVES TO TREAT PATHOLOGIES OR DYSFUNCTIONS OF THE MITOCHONDRIA

[0002] TECHNICAL FIELD

[0003] The present invention aims to provide new pharmacological tools for treating mitochondrial diseases or dysfunctions, in particular those associated with mitochondrial complex I deficiencies.

[0004] PRIOR STATE OF THE ART

[0005] Mitochondrial diseases are common metabolic disorders characterized by high clinical and genetic heterogeneity, resulting in mitochondria failing to produce enough energy to allow the body to function properly. Mitochondrial diseases can be present at birth but can also appear at any age. Genetically, mitochondria have the particularity of possessing their own DNA (or mitochondrial DNA), but the majority of mitochondrial proteins are dependent on the nuclear genome. Thus, both genetic abnormalities carried by the mitochondrial and nuclear genomes are responsible for mitochondrial diseases, reflecting the very high clinical and genetic heterogeneity. It is estimated that one in 4,300 people is affected by a mitochondrial disease (Gorman et al., 2015, Ann Neurol. 77(5):753-9).

[0006] Mitochondrial diseases can affect almost any part of the body, including cells in the brain, nerves, muscles, kidneys, heart, liver, eyes, ears, or pancreas. Symptoms of mitochondrial diseases depend on the organs affected, preferentially affecting energy-intensive tissues such as muscles, brain, and heart. Patients' symptoms can be moderate to severe and involve one or more organs.

[0007] Symptoms of mitochondrial diseases may include:

[0008] - poor growth

[0009] - muscle weakness, muscle pain, low muscle tone, exercise intolerance

[0010] - vision and / or hearing problems - learning difficulties, developmental delays, mental retardation

[0011] - autism, or characteristics of autism

[0012] - cardiac dysfunction, cardiac arrhythmia or cardiac conduction abnormalities

[0013] - liver or kidney diseases

[0014] - gastrointestinal disturbances, difficulty swallowing, diarrhea or constipation, unexplained vomiting, cramps, gastrointestinal reflux

[0015] - diabetes

[0016] - an increased risk of infections

[0017] - neurological problems, epileptic seizures, migraines, strokes

[0018] - mobility disorders

[0019] - thyroid and / or adrenal gland dysfunction

[0020] - respiratory problems

[0021] - lactic acidosis, i.e. an accumulation of lactate in the blood or urine

[0022] - dementia.

[0023] Mitochondrial dysfunction can also occur when mitochondria do not function properly, possibly due to another disease or condition. Many conditions can lead to secondary mitochondrial dysfunction, including Alzheimer's or Parkinson's disease, muscular dystrophies, Lou Gehrig's disease, diabetes, and cancer. People with secondary mitochondrial dysfunction do not have a primary mitochondrial disease of genetic origin but experience similar symptoms. In addition, certain medications can target and damage mitochondria.

[0024] Mitochondrial complex I deficiency is the most common defect observed in more than 30% of mitochondrial diseases. Among these, the two most frequent clinical phenotypes related to complex I deficiencies are Leigh syndrome, which is often fatal, or milder phenotypes such as Leber hereditary optic neuropathy (LHON). MELAS syndrome has also been considered a common mitochondrial disease due to mutations in the mitochondrial genome and associated with a severe reduction in mitochondrial complex I activity. Complex I is composed of at least 44 subunits, seven of which, namely ND1 to 6 and ND4L, are encoded by mitochondrial genes, while the others are encoded by nuclear genes. Therefore, the clinical and molecular features associated with inherited complex I deficiency are highly variable.Among these complex I subunits, mutations targeting the NDUFV1 gene have been shown to cause severe neurological phenotypes (Schuelke et al., 1999, Nat Genet. 21(3):260-61). Mutations affecting the NDUFS8 subunit have been associated with Leigh syndrome (Procaccio et al., 2004, Neurology 62:1899), and mutations targeting the mitochondrial DNA-encoded ND3 subunit have been reported in Leigh syndrome and LHON (Sarzi et al., 2007 American Journal of Medical Genetics 143:33-41; Wang et al., 2009, Neurogenetics. 10:337-345). Furthermore, mutations affecting the mitochondrial DNA-encoded ND6 subunit have been reported in NHOL (Johns et al., 1992, Biochem Biophys Res Commun. 187(3): 1551-7).

[0025] Furthermore, complex I deficiency has been identified in secondary mitochondrial dysfunction associated with age-related neurodegenerative diseases, such as Parkinson's disease.

[0026] Although most mitochondrial diseases are genetic in origin, gene therapy appears difficult to implement due to the genetic and clinical diversity and complexity of these diseases.

[0027] The goal of current treatments is to improve symptoms and slow the progression of the disease or dysfunction with, for example, the following recommendations:

[0028] - use of vitamin therapy

[0029] - energy conservation

[0030] - slowdown in activities

[0031] - maintaining the environment at room temperature

[0032] - protection against exposure to intercurrent diseases (infections, stress)

[0033] - maintaining adequate nutrition and hydration.

[0034] It should be noted that documents WO2020 / 254632 and W02022 / 018297 have recently reported the management of these pathologies or deficiencies using disulfiram and alverine, respectively. However, there remains a need to find new therapeutic and pharmacological approaches to treat this type of dysfunction or disease, based on causal and not symptomatic treatment.

[0035] Document WO2014 / 150688 describes the treatment of cancer using molecules capable of inhibiting enzymes of the mitochondrial pathway (1-C) such as ebselen, in particular the enzyme MTHFD2.

[0036] JIA ZHI-QIANG et al. (Neurosc. Letters, 2018, vol. 678, pp. 110-17) report the beneficial effect of ebselen in the management of acute spinal cord injury, with a neuroprotective effect and improvement of mitochondrial function.

[0037] The paper ARAKAWA MOTOKI et al. (Cerebellum, 2007, 6(4), pp 308-14) deals with neurodegenerative diseases such as Alzheimer's or Parkinson's disease, described as associated with mitochondrial alterations. This paper focuses on the positive effect of NAC (N-acetylcysteine) against 4-hydroxynonenal (HNE)-induced neuron toxicity, and mentions a neuroprotective effect of ebselen and its possible use in combination with NAC.

[0038] The paper AZAD GAJENDRA et al. (Molecular Biology Reports, 2014, 41(8), pp 4865-79) is a review dedicated to ebselen, described as a promising antioxidant drug. Table 2 lists all the pathologies that can be improved by the administration of ebselen.

[0039] The paper by CAPPER MICHAEL et al. (Nature Communications, 2018, 9(1)) focuses on the effect of ebselen on the enzyme superoxide dismutase 1 (SOD1), mutations in which can lead to conditions such as amyotrophic lateral sclerosis (ALS).

[0040] DETAILED DESCRIPTION OF THE INVENTION

[0041] The inventors have shown that ebselen, a drug currently used in clinical trials for the treatment of hearing disorders such as Ménière's disease, but also its derivatives, such as ethasalen, were active in the context of the treatment of diseases associated with mitochondrial dysfunction, in particular diseases of the mitochondrial respiratory chain, advantageously associated with a deficiency of complex I. Thus, this work opens the way to the use of a new family of compounds in this context.

[0042] Definitions

[0043] The definitions below give the meaning generally used within the scope of the invention and must be taken into account unless another definition is explicitly indicated.

[0044] The terms "about", "roughly", "in the order of" or "approximately" used herein to denote a measurable value such as a quantity, a time and the like, should be understood to encompass variations of ± 20% or ± 10%, preferably ± 5%, more preferably ± 1%, and even more preferably ± 0.1% from the specified value.

[0045] Intervals / Ranges: Throughout this disclosure, various aspects of the invention may be presented in the form of an interval of values ​​(range format). It should be understood that the description of values ​​in the form of an interval is for convenience and brevity only and should not be construed as limiting the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all possible subranges as well as the individual numerical values ​​within that range. For example, the description of a range such as “from 1 to 6” should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc. as well as individual numbers within that range, e.g. 1, 2, 2, 7, 3, 4, 5, 5, 3 and 6. This applies regardless of the extent of the range.

[0046] “Isolated” means extracted or removed from its natural environment or state. For example, an isolated nucleic acid or peptide is a nucleic acid or peptide that has been extracted from the natural environment in which it is usually found, whether in a living plant or animal, for example. A nucleic acid or peptide, for example, that is naturally present in a living animal is not an isolated nucleic acid or peptide for the purposes of the invention, whereas the same nucleic acid or peptide partially or completely separated from other components present in its natural environment is itself “isolated” for the purposes of the invention. An isolated nucleic acid or protein may exist in a substantially purified form, or may exist in a non-native environment such as, for example, a host cell.The term “abnormal,” when used in the context of organisms, tissues, cells, or components thereof, means 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 organisms, tissues, cells, or components thereof that exhibit the respective “normal” (expected) characteristic. Characteristics that are normal or expected for one type of cell or tissue may be abnormal for another type of cell or tissue.

[0047] The terms "patient," "subject," "individual," and the like are used interchangeably herein and refer to any animal or cell thereof, in vitro or in situ, that can be subjected to the methods described herein. In certain non-limiting embodiments, the patient, subject, or individual is an animal, preferably a mammal, more preferably a human, either female or male. It may also be a mouse, rat, pig, dog, or non-human primate (NHP), such as a macaque monkey.

[0048] For the purposes of the invention, a “disease” or “pathology” is a state of health of an animal in which its homeostasis is negatively affected and which, if the disease is not treated, continues to deteriorate. Conversely, for the purposes of the invention, a “disorder” or “dysfunction” is a state of health in which the animal is able to maintain its homeostasis but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. In the absence of treatment, a disorder does not necessarily lead to a deterioration in the animal’s state of health over time.

[0049] A disease or disorder is “mitigated” (“reduced”) or “improved” if the severity of a symptom of the disease or disorder, the frequency with which that symptom is experienced by the subject, or both, are reduced. This also includes the cessation of disease progression, i.e., the cessation of progression of the disease or disorder. A disease or disorder is “cured” (“recovered”) if the severity of a symptom of the disease or disorder, the frequency with which such a symptom is experienced by the patient, or both, are eliminated. In the context of the invention, a “therapeutic” treatment is a treatment administered to a subject who exhibits the symptoms (signs) of a pathology, with the aim of reducing or eliminating those symptoms. In the context of the invention, “treatment of a disease or disorder” means reducing the frequency or severity of at least one sign or symptom of a disease or disorder experienced by the subject.A treatment is said to be prophylactic when it is administered to prevent the development, spread or worsening of a disease, particularly if the subject does not or does not yet present the symptoms of the disease and / or for which the disease has not been diagnosed.

[0050] As used herein, “treating a disease or disorder” means reducing the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject. Disease and disorder are used interchangeably in the context of the treatment of the invention.

[0051] For the purposes of the invention, an “effective amount” or “effective amount” of a compound is that amount of compound that is sufficient to provide a beneficial effect to the subject to whom the compound is administered. The term “therapeutically effective amount” refers to an amount that is sufficient or effective to prevent or treat (in other words, delay or prevent the development of, prevent the progression of, inhibit, reduce, diminish, or reverse) a disease or disorder, including the alleviation of the symptoms of that disease or disorder.

[0052] The present invention therefore relates to the use of a compound having a group of formula: or a compound of formula: , , , , SeO (Se=O) each of the phenyl rings A and B is optionally substituted by one or more substituents, wherein each substituent is independently selected from:

[0053] - a halogen, which is preferably selected from F, Cl and Br

[0054] - an alcohol

[0055] - an amine

[0056] - a nitro

[0057] - a C1-C4 alkyl, such as a C1-C2 alkyl or a Cl alkyl, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0058] - a C1-C4 alkoxy, such as a C1-C2 alkoxy or a Cl alkoxy, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br or a pharmaceutical composition containing it, for the treatment of a disease associated with mitochondrial dysfunction, or for the preparation of a medicament for the treatment of a disease associated with mitochondrial dysfunction. In other words, the invention relates to a compound having a group of formula: in which

[0059] E is O, S, SO (S=O), SO2(O=S=O), Se or SeO (Se=O) each of the phenyl rings A and B is optionally substituted by one or more substituents, wherein each substituent is independently selected from:

[0060] - a halogen, which is preferably selected from F, Cl and Br

[0061] - an alcohol

[0062] - an amine

[0063] - a nitro

[0064] - a C1-C4 alkyl, such as a C1-C2 alkyl or a Cl alkyl, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0065] - a C1-C4 alkoxy, such as a C1-C2 alkoxy or a Cl alkoxy, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br or a pharmaceutical composition containing it, for use in the treatment of a disease associated with mitochondrial dysfunction. According to another aspect, the invention relates to a method of treating a disease associated with mitochondrial dysfunction comprising administering a compound having a group of formula: or formula in which

[0066] E is O, S, SO (S=O), SO2(O=S=O), Se or SeO (Se=O) each of the phenyl rings A and B is optionally substituted by one or more substituents, wherein each substituent is independently selected from:

[0067] - a halogen, which is preferably selected from F, Cl and Br

[0068] - an alcohol

[0069] - an amine

[0070] - a nitro

[0071] - a C1-C4 alkyl, such as a C1-C2 alkyl or a Cl alkyl, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0072] - a C1-C4 alkoxy, such as a C1-C2 alkoxy or a Cl alkoxy, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br or a pharmaceutical composition comprising such a compound, in a subject.

[0073] A compound used in the context of the invention may exhibit inositol monophosphatase (IMPase) inhibitory activity, said activity being able to be tested as described by Singh et al. (Nat Commun. 2013;4:1332. doi:10.1038 / ncomms2320). According to a particular embodiment, such a compound is not valproic acid or carbamazepine.

[0074] A compound used in the context of the invention may exhibit antioxidant activity, said activity being able to be tested as described by Noguchi et al. (Biochem Pharmacol 1992; 44: 39-44) and Nakamura et al. (J Biol Chem., 2002; 277(4):2687-94).

[0075] According to a particular embodiment, such a compound is not ascorbic acid.

[0076] According to a first particular embodiment, a compound used within the framework of the invention having a group of formula: is ethaselen or one of its derivatives or analogues, as defined below.

[0077] Ethaselen or BBSKE (CAS No.: 217798-39-5) has the following formula:

[0078] A derivative or analogue of ethaselen is, for example, the chlorinated derivative MAD423 of formula:

[0079] Thus, a compound of interest within the framework of the invention has the following formula:

[0080] E is O, S, SO, SO2, Se or SeO each of the phenyl rings A and B is optionally substituted by one or more substituents, wherein each substituent is independently selected from:

[0081] - a halogen, which is preferably selected from F, Cl and Br

[0082] - an alcohol

[0083] - an amine

[0084] - a nitro

[0085] - a C1-C4 alkyl, such as a C1-C2 alkyl or a Cl alkyl, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0086] - a C1-C4 alkoxy, such as a C1-C2 alkoxy or a Cl alkoxy, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br.

[0087] According to another embodiment, a compound used in the context of the invention is as described in document WO2012 / 107735, namely a compound of formula I, or a pharmaceutically acceptable salt thereof, in which formula I is: in which

[0088] E is O, S, SO, SO2, Se or SeO each of the phenyl rings A and B is optionally substituted by one or more substituents, wherein each substituent is independently selected from:

[0089] - a halogen, which is preferably selected from F, Cl and Br

[0090] - an alcohol

[0091] - an amine

[0092] - a nitro

[0093] - a C1-C4 alkyl, such as a C1-C2 alkyl or a Cl alkyl, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0094] - a C1-C4 alkoxy, such as a C1-C2 alkoxy or a Cl alkoxy, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br.

[0095] According to a preferred embodiment, such a compound is:

[0096] - 2-phenyl-1,2-benzisoselenazol-3(2H)-one or ebselen of formula:

[0097] - 2-phenyl-l,2-benzisothiazol-3(2H)-one or ebselen sulfur (2-phenyl-2,3-dihydro-l,2-benzothiazol-3-one) of formula:

[0098] - l-oxyde-2-phenyl-l,2-benzisoselenazol-3(2H)-one or ebselen oxide (ebselen oxide) or ebselen selenoxide of formula:

[0099] - ebselen -OH ortho of formula: - MAD309 (6-hydroxy-2-phenylbenzo[d][l,2]selenazol-3(2H)-one 1-oxide) of

[0100] - MAD331 (6-chloro-2-phenylbenzo[d][l,2]selenazol-3(2H)-one) of formula:

[0101] - MAD349 (2-(2,6-dichlorophenylbenzo[d][l,2]selenazol-3(2H)-one) of formula:

[0102] - MAD383 (6-chloro-2-(3-trifluoromethyl)phenyl)benzo[d][l,2]selenazol-3(2H)- one) of formula:

[0103] - MAD385 (2-(3-difluoromethoxy)phenyl)benzo[d][l,2]selenazol-3(2H)-one) of formula: - MAD413 (6-chloro-2-phenylbenzo[d][l,2]selenazol-3(2H)-one 1-oxide) of formula:

[0104] Preferred compounds according to the invention are ebselen and ebselen oxide, advantageously ebselen, and their derivatives or analogues as defined below.

[0105] Ebselen (2-phenyl-1,2-benzoselenazol-3(2H)-one; CAS number: 60940-34-3), also known as PZ 51, DR3305, and SPI-1005, is a synthetic molecule capable of mimicking glutathione peroxidase. Therapeutic activity of this molecule has been described in relation to ischemic lesions, cardiovascular events, hearing loss, and bipolar disorders. It may also be effective in treating Clostridioides difficile infections and exhibits antifungal activity against Aspergillus fumigatus.

[0106] In practice, this molecule is the subject of phase III clinical trials for the treatment of Menière's disease, a hearing disorder. Ebselen is administered orally at different doses from 200 mg to 600 mg (in the form of 200 mg capsules) per day for at least 21 days.

[0107] Also covered by the present invention are derivatives or analogues of these compounds, in particular ebselen, having the same biological activity, namely that reported in the examples, in particular the restoration of respiratory growth, cellular respiration or mitochondrial ATP level, both in yeast models, in particular Saccharomyces cerevisiae, filamentous fungus, in particular Podospora anserina, and on human cells, in particular fibroblasts or myoblasts of patients suffering from these pathologies. The term "derivatives" (or "analogues") includes derivatives and metabolites, as well as pharmaceutically acceptable salts. A derivative is a compound originating from another (the precursor, with a typically similar chemical structure) after transformation of the latter. The derivative may differ by one or more atoms or functional groups.A metabolite is a stable intermediate compound or a compound resulting from the biochemical transformation of an initial molecule by metabolism.

[0108] By "pharmaceutically acceptable salts" is meant the addition salts of the compound, which can be obtained by reacting this compound with a mineral or organic acid according to a method known per se. Among the acids conventionally used for this purpose, mention may be made of hydrochloric, hydrobromic, sulfuric, phosphoric, 4-toluene sulfonic, methane sulfonic, cyclohexyl sulfamic, oxalic, succinic, formic, fumaric, maleic, citric, aspartic, cinnamic, lactic, glutamic, N-acetyl-aspartic, N-acetyl-glutamic, ascorbic, malic, benzoic, nicotinic and acetic acids. According to a preferred embodiment, the compound is not in the form of a salt.

[0109] Such compounds, including ebselen, may be modified to increase their stability, bioavailability and / or ability to reach target tissues, including mitochondria.

[0110] As known to those skilled in the art, said compounds, in particular ebselen, may be present in the composition in a naked (free) form or contained in delivery systems which increase stability, targeting and / or bioavailability, such as liposomes, or incorporated into supports such as hydrogels, cyclodextrins, biodegradable nanocapsules, bioadhesive microspheres, vectors or in combination with a cationic peptide.

[0111] The present invention also relates to pharmaceutical compositions containing as active ingredient at least one compound as defined above, as well as the use of this compound or this composition as a medicament or medicinal product. Thus, the present invention relates to pharmaceutical compositions comprising a compound according to the invention for the intended use. Advantageously, these compositions comprise a therapeutically effective amount of said compound, and a pharmaceutically acceptable carrier. In a particular embodiment, the term "pharmaceutically acceptable" means approved by a regulatory body of the federal or state government or listed in the American or European Pharmacopoeia or in another pharmacopoeia generally recognized for use in animals and humans.The term "carrier" means a diluent, adjuvant, excipient, or vehicle with which the therapeutic product is administered. These pharmaceutical carriers may 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 others. Saline solutions and aqueous solutions of dextrose and glycerol may also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerol, propylene glycol, water, ethanol, and others.

[0112] The composition, if any, may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions may take the form of solutions, suspensions, emulsions, sustained-release formulations, and the like. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin. These compositions contain a therapeutically effective amount of the therapeutic agent, preferably in purified form, together with an appropriate amount of carrier so as to provide the form for adequate administration to the subject.

[0113] In a preferred embodiment, the composition is formulated in accordance with routine procedures as a pharmaceutical composition suitable, for example, for oral administration to humans. Typically, compositions for oral administration are in the form of tablets, optionally scored tablets or effervescent tablets, further containing excipients suitable for the solid dosage form and for administration to humans. For example, ebselen may be in powder form packaged in tablets which may contain 200 mg of the active ingredient. These tablets may be crushed and mixed with liquids. Alternatively, the composition may be in a liquid form, advantageously an aqueous composition. Any other suitable solvent may be used.

[0114] The amount of the therapeutic agent of the invention, i.e., a compound as described above, that will be effective in the treatment of a disease can be determined by standard clinical techniques. In addition, in vivo and / or in vitro assays, such as those described in the examples below, may optionally be used to help predict optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, the weight and severity of the disease, and should be decided according to the judgment of the practitioner and the conditions of the individual patient.

[0115] According to a particular embodiment, the composition of the invention is in a solid form, advantageously a tablet, which can contain 200 mg of the active compound, in particular ebselen.

[0116] According to one embodiment, the composition of the invention is in a liquid form and advantageously comprises less than 30 pM, 20 pM or even 15 pM of the active compound, in particular ebselen. According to another embodiment, the composition of the invention is in a liquid form and advantageously comprises 1 nM or more, or even 10 nM or more, 30 nM or more, or even 100 nM or more, with for example an optimal concentration of 1 pM of the active compound, in particular ebselen.

[0117] An appropriate route of administration should allow delivery of a therapeutically effective amount of the therapeutic product to the target tissues, depending on the disease.

[0118] Ebselen and its derivatives may be administered in a pharmaceutically acceptable form by one of the routes of administration known for this type of active ingredient. The available routes of administration are: topical (local), enteral (systemic action, but delivered through the gastrointestinal (GI) tract), or parenteral (systemic action, but delivered through routes other than the GI tract). In the specific case of mitochondrial diseases, the preferred route of administration of the compositions disclosed herein is generally enteral, which includes oral, sublingual, buccal, and preferably oral administration. In other embodiments, it may be parenteral administration, including intramuscular (i.e., into the muscle) or systemic (i.e., into the circulating system) administration.In this context, the term "injection" (or "infusion" or "infusion") encompasses intravascular administration, particularly intravenous (IV), and intramuscular (IM). Injections are usually given using syringes or catheters.

[0119] According to one embodiment, the composition is administered orally, intramuscularly, intraperitoneally, subcutaneously, topically, locally or intravascularly, advantageously orally. According to a preferred embodiment, the composition is intended for oral administration.

[0120] A composition according to the invention is preferably presented in a solid galenic form suitable for oral administration, advantageously in the form of one or more gelatin capsules, capsules or tablets.

[0121] Alternatively, they may be liquid preparations such as elixirs and suspensions containing various masking substances for coloring, flavoring, and stabilization.

[0122] According to a preferred embodiment, the composition is intended for oral administration. Advantageously, the composition is administered per os, that is to say by mouth.

[0123] Preferably, a composition according to the invention is administered orally, in particular in the form of capsules, capsules or tablets.

[0124] To produce the oral dosage forms according to the invention, in particular capsules, the active substance can be mixed with various 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 necessary.

[0125] It may, in certain cases, be advantageous to provide controlled-release forms, in particular prolonged-release forms using known galenic forms. Similarly, a composition according to the invention may be intended for the preparation of a pharmaceutical composition which can be administered by injection.

[0126] The pharmaceutical composition according to the invention can be dissolved or suspended in a sterile, pharmaceutically acceptable injectable liquid, such as sterile water, a sterile organic solvent or a mixture of these two liquids for intravenous administration.

[0127] 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.

[0128] As already mentioned, a composition according to the invention is preferably in a solid galenic form suitable for oral administration, advantageously in the form of one or more gelatin capsules, capsules or tablets.

[0129] These can be taken with a little water before or during the main meal. According to a preferred embodiment, the composition according to the invention is administered daily, for example once a day, or even two or more times a day. The treatment can last several weeks, several months, several years or even a lifetime.

[0130] In general, the dosage of the therapeutic agent, i.e., ebselen or one of its derivatives, varies depending on factors such as the subject's age, weight, height, sex, general health, and medical history. Typically, it is desirable to provide the patient with an individual dose of the therapeutic agent that is effective but not toxic.

[0131] According to a particular embodiment of the invention, the dose of the composition, advantageously the daily dose to be taken orally by a human, is less than or equal to 10 mg / kg or 9, 8, 7, 6, 5, 4, 3 mg / kg, or even less than or equal to 2.5, 2, 1.5 or 1 mg / kg, or even less than or equal to 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3 or 0.1 mg / kg. As already indicated, the patient is advantageously a human, in particular a newborn, a young child, a child, an adolescent or an adult, regardless of their sex. The therapeutic tool according to the invention may however be adapted and useful for the treatment of other animals, in particular pigs, mice, dogs or macaque monkeys.

[0132] Generally, the present invention relates to the treatment of mitochondrial diseases in general, i.e. diseases related to or caused by mitochondrial dysfunction. In the context of the present application, the expression "disease associated with mitochondrial dysfunction" is used to encompass all of these situations.

[0133] Related to the examples below showing a positive effect of ebselen or one of its derivatives on the mitochondrial respiratory chain, diseases of particular interest are mitochondrial respiratory chain diseases.

[0134] Several mitochondrial diseases have been documented in the prior art:

[0135] NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome induces a variable combination of developmental delay, retinitis pigmentosa, dementia, seizures, ataxia, proximal neurogenic muscle weakness, and sensory neuropathy. This NARP syndrome is caused by various mutations in the mitochondrial ATP6 gene, which encodes the α-subunit of ATPase (complex V of the OXPHOS system). Mutations are often heteroplasmic (coexistence of mutated and wild-type (WT) mitochondrial DNA (mtDNA) in the same cell). Depending on the type of mutation and the percentage of mutated mtDNA (degree of heteroplasmy), the clinical consequences are more or less severe. ATP6 m.8993T>C / G mutations are among the most frequent in NARP patients and result in severe forms of NARP syndrome.FMC1 is a nuclear gene that encodes a protein required at high temperature (35-37°C) for the assembly of the F1 sector of ATP synthase, thus mimicking the heteroplasmy observed in NARP patients. Indeed, when cultured at a restrictive temperature (35-37°C), mitochondria of the fmclA mutant contain significantly fewer assembled ATP synthase complexes than a wild-type (WT) strain, but those that assemble are fully functional. This heterogeneity is also found in patients with reduced ATP synthase levels due to heteroplasmic mutations in the ATP6 gene. Therefore, the fmclA mutant constitutes a suitable model for this disease, particularly the equivalent of the m.8993T mutant <G (MR14, NARP) (Schon EA et al. (2001) Cell Dev Biol. 12(6):441-8).

[0136] The TAZ gene encodes tafazzin, a mitochondrial transacylase that catalyzes the remodeling of immature cardiolipin into its mature composition containing a predominance of tetralinoleoyl moieties. Mutations in TAZ result in Barth syndrome, an X-linked disorder classically characterized by dilated cardiomyopathy (DCM) with endocardial fibroelastosis (EFE), predominantly proximal skeletal myopathy, growth retardation, neutropenia, and organic aciduria, including excess 3-methylglutaconic acid (Barth, PG et al. (1996) J Inherit Metab Dis, 19, 157-160).

[0137] The COX2 and SURF1 genes encode a subunit and an assembly factor of mitochondrial complex IV, respectively. Mutations are associated with Leigh syndrome, a severe, progressive neurodegenerative disease that manifests in the first months or years of life and is responsible for early death. Affected individuals typically present with global developmental delay or developmental regression, hypotonia, ataxia, dystonia, and ophthalmologic abnormalities, such as nystagmus or optic atrophy (Barrientos, A. et al. (2002) EMBO J, 21, 43-52).

[0138] MPV17 encodes a mitochondrial inner membrane protein of unknown function. MPV17 Q mutations result in:

[0139] - Mitochondrial DNA depletion syndrome-6, an autosomal recessive disease characterized by infantile onset of progressive liver failure, often leading to death within the first year of life. Those who survive develop progressive neurological impairment, including ataxia, hypotonia, dystonia, and psychomotor regression (Spinazzola, A. et al. (2006) Nat Genet, 38, 570-575).

[0140] - Navajo-type neuropathy: Manifestations include severe anesthesia leading to corneal ulceration, painless fractures and acral mutilation, muscle weakness, absent or markedly diminished deep tendon reflexes, without intellectual deficit (Karadimas, CL et al. (2006) Am J Hum Genet, 79, 544-548).Of particular interest is the treatment of a disease selected from the group consisting of: MELAS syndrome, maternally inherited myopathy and cardiomyopathy, NARP or MILS syndrome, Leigh syndrome, Leber hereditary optic neuropathy (LHON), Barth syndrome, mitochondrial DNA depletion syndrome including 4A (Alpers type) and 4B (MNGIE type), mitochondrial recessive ataxia syndrome, ataxic sensory neuropathy, dysarthria and ophthalmoplegia, spinocerebellar ataxia with epilepsy, progressive external ophthalmoplegia, mitochondrial DNA depletion syndrome-6, Navajo-type neuropathy, Behr syndrome, mitochondrial DNA depletion syndrome-14, infantile cardioencephalomyopathy due to cytochrome c oxidase deficiency (COA6 mutations), nuclear deficiency of the mitochondrial complex III type 1, GRACILE syndrome and Bjomstad syndrome.

[0141] Of particular interest is the treatment of diseases associated with mitochondrial complex I deficiency(ies). Some diseases are solely related to complex I dysfunction, while other diseases are associated with multiple deficiencies, e.g., affecting several mitochondrial complexes.

[0142] The respiratory chain in mitochondria is known to be involved in oxidative phosphorylation, an important cellular process that uses oxygen and simple sugars to create adenosine triphosphate (ATP), the cell's primary energy source. Five protein complexes, the OXPHOS system, each composed of several proteins, are involved in this process. These complexes are named complex I, complex II, complex III, complex IV, and complex V, respectively. Complex I (CI or NADH dehydrogenase or NADH coenzyme Q reductase), the first enzyme in the respiratory chain, is a very large protein complex (approximately 1000 kDa) composed of at least 44 subunits, 7 of which are encoded by mitochondrial DNA (ND1 to ND6 and ND4L).According to a particular embodiment, a compound according to the invention, in particular ebselen, can be used to treat a so-called "primary" mitochondrial disease, i.e. due to a genetic anomaly identified in at least one subunit of the OXPHOS complexes (in particular complex I) linked to one or more mutations of the mitochondrial or nuclear DNA. These pathologies are associated with neurological, cardio-muscular or ophthalmological symptoms, linked to the tissues or organs most affected in these mitochondrial diseases, even if other organs or tissues are possibly affected. According to another particular embodiment, a compound according to the invention, in particular ebselen, can be used to treat a so-called "secondary" mitochondrial disease.In this case, the genetic abnormality does not directly involve the OXPHOS complexes, but the pathology will affect mitochondrial functions and, in particular, may lead to a reduction in the enzymatic activity of this process. Such a disease can also be due to non-genetic causes such as exposure to environmental factors or aging. This is particularly the case with Parkinson's disease or other age-related neurodegenerative disorders.

[0143] According to one embodiment, the mitochondrial deficiency or dysfunction results from a genetic disease.

[0144] Genetic diseases are, by definition, diseases resulting from one or more genetic defects (abnormalities) (or mutations) in one or more genes. Genetic defects can affect mitochondrial DNA and / or nuclear genes. The genetic defects responsible for mitochondrial diseases can be point mutations, resulting in a codon change. However, the diseases can be related to the deletion or insertion of one or more bases or codons.

[0145] According to a specific embodiment, the disease results from one or more genetic deficiencies (or mutations) in one or more genes involved in the functionality of OXPHOS complexes, in particular complex I.

[0146] A non-exhaustive list of such genes includes:

[0147] - the structural genes of complex I, in particular MTND1 (or ND / ), MTND2 (or ND2 MTND3 (or ND 3), MTND4 (or ND4), MTND5 (or ND5), MTND6 (or ND6), MTND4L (or ND4E), 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

[0148] - the assembly genes of complex I, in particular NDUFAF1, NDUFAF2, NDUFAF3, NDUFAF4, NDUFAF5, NDUFAF6, NDUFAF7, NDUFAF8, NUBPL, ACAD9, TMEM70, TMEM126B, FOXRED1, ECSIT, AIF, TIA4MDC1. According to a particular embodiment, the diseases to be treated within the scope of the invention are linked to or due to at least one genetic defect or mutations in at least one of the following genes: MTTL1, ATP6, FMC1, TAZ, COX2, SURF1, POLG, MPV17, OPA1, COA6, ND6 and BCS1L, advantageously ATP6, FMC1, TAZ, COX2, SURF1 or MPV17.

[0149] Several mitochondrial diseases of genetic origin, particularly related to mitochondrial complex I deficiencies, have been documented in the prior art: LHON syndrome or Leber hereditary optic neuropathy generally appears in young adults. The onset is abrupt with a rapid decline in vision in the center of the eye, corresponding to a decrease in central visual acuity. Most often, a peripheral visual field persists, like a halo of vision around a blind area. This disease is due to commonly homoplasmic mutations in the genes coding for the subunits of complex I of the respiratory chain. In practice, the mitochondrial DNA mutations m.H778G>A, m.3460G>A and m,14484T>C represent approximately 95% of LHON mutations.

[0150] Leigh syndrome (or LS) is a severe, progressive neurodegenerative disorder. Affected individuals typically present with global developmental delay or regression, hypotonia, ataxia, dystonia, and ophthalmologic abnormalities, such as nystagmus or optic atrophy. Leigh syndrome can also have adverse multisystemic effects on cardiac, hepatic, gastrointestinal, and renal organs. Biochemical studies in patients with Leigh syndrome tend to show increased lactate and abnormalities in mitochondrial oxidative phosphorylation. Leigh syndrome may be associated with mutations in genes encoding complex I subunits, such as NDUFV1 mutations or the MTND5 m.13513 G>A mutation.

[0151] MELAS syndrome, comprising mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes, is a genetically heterogeneous mitochondrial disease with a variable clinical phenotype. This disorder presents with features of central nervous system involvement, including seizures, hemiparesis, hemianopia, cortical blindness, and episodic vomiting. This syndrome was first associated with the m.3243A>G mutation in mitochondrial DNA, i.e., the tRNA gene. Leu (UUR) (MTTLl), which induces impaired translation of complex I mRNA into proteins and thus a reduction in the amount of complex I structural proteins such as the mitochondrial subunit ND6. MELAS syndrome can also be associated with other mitochondrial DNA mutations such as the m.3260A>G mutation, which also affects tRNA Leu (UUR)'This m.3260A>G mutation can also result in other clinical phenotypes, including maternally derived myopathy and cardiomyopathy.

[0152] Thus, the treatment of genetic diseases shown to be associated with complex I deficiency, such as MELAS syndrome, Leigh syndrome, and Leber hereditary optic neuropathy (LHON), is of particular interest. It should be noted that these diseases may be associated with other symptoms such as cardiac, myopathic, or neurological clinical phenotypes.

[0153] More generally, a compound according to the invention, in particular ebselen, can be used to treat mitochondrial dysfunction, in particular mitochondrial dysfunction associated with complex I deficiencies. Mitochondrial dysfunction, characterized by a loss of efficiency of the electron transport chain and reductions in the synthesis of high-energy molecules such as adenosine-5'-triphosphate (ATP), is a characteristic of aging and generally of all chronic diseases.

[0154] These diseases include:

[0155] - neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (Lou Gehrig's disease) and Friedreich's ataxia;

[0156] - cardiovascular diseases, such as atherosclerosis and other heart and vascular conditions;

[0157] - diabetes and metabolic syndrome;

[0158] - autoimmune diseases, such as multiple sclerosis, systemic lupus erythematosus and type 1 diabetes;

[0159] - neurobehavioral and psychiatric illnesses, such as autism spectrum disorders, schizophrenia, bipolar and mood disorders;

[0160] - gastrointestinal disorders;

[0161] - fatigue diseases, such as chronic fatigue syndrome and Gulf War illness; musculoskeletal diseases, such as fibromyalgia and skeletal muscle hypertrophy / atrophy; - muscular dystrophies,

[0162] - cancer; and

[0163] - chronic infections.

[0164] The treatment recommended by the present invention, based on ebselen or one of its derivatives, can also be combined with other treatments, intended to treat the same pathology or the same condition, or another disease.

[0165] According to a particular embodiment, this other treatment is based on the administration of another compound of interest.

[0166] Thus and according to this aspect, the composition according to the invention is combined with at least one other 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 to take into account their particularities and in particular their bioavailability.

[0167] According to a particular embodiment, the present invention relates to a composition, advantageously a pharmaceutical composition or a medicament containing a compound as described above and potentially other active molecules (other gene therapy molecules, chemical groups, peptides or proteins, etc.) for the treatment of the same disease or a different disease, advantageously the same disease.

[0168] Preferably, the pharmaceutical composition according to the present invention and at least one compound for the treatment of the same disease or a different disease are administered simultaneously, separately or in a time-spaced manner to treat the same disease or a different disease.

[0169] More generally, with regard to mitochondrial diseases, another compound capable of improving mitochondrial function can be administered simultaneously or at different times. In the case of simultaneous administration, the two compounds can be combined in the same composition.

[0170] Examples of such additional compounds are natural supplements, such as L-carnitine, alpha-lipoic acid (α-lipoic acid [l,2-dithiolane-3-pentanoic acid]), coenzyme Q10 (CoQ10 or ubiquinone), riboflavin (vitamin B2) reduced to nicotinamide adenine dinucleotide (NADH), L-arginine, possibly in combination.

[0171] Examples of compounds used, for example, in MELAS syndrome are nitric oxide (NO) precursors such as L-arginine and citrulline.

[0172] According to a particular embodiment and in view of the examples demonstrating a potential synergistic effect, a compound according to the invention, in particular ebselen, is combined with one of the compounds described in documents WO2020 / 254632 and WO2022 / 018297, advantageously chosen from the group consisting of: disulfiram, sodium diethyldithocarbamate, alverine and alverine citrate.

[0173] Subjects who may benefit from the compositions of the invention include all patients having a disease associated with mitochondrial dysfunction, in particular mitochondrial dysfunction associated with complex I deficiencies, diagnosed as having such a disease or at risk of developing such a disease.

[0174] A subject to be treated with a composition according to the invention may be selected on the basis of various criteria. With regard to mitochondrial dysfunction, in particular complex I deficiencies, several tests may be carried out, for example:

[0175] - At the biochemical level: from a biopsy, particularly muscular or cutaneous of the subject, we can measure the consumption of CL and / or the activity of mitochondrial complex I. The activity of the other complexes of the respiratory chain can also be evaluated to determine if the mitochondrial dysfunction is solely due to deficiencies of complex I;

[0176] - At the genetic level: sequencing of mitochondrial or nuclear DNA extracted from blood, cells or a biopsy sample, for example from the skin, makes it possible to identify one or more molecular abnormalities, in particular mutations or deletions / insertions in the genes listed above. Alternatively, the expression or activity of the corresponding proteins can be evaluated by any method known to those skilled in the art (for example by western blot).

[0177] One objective of the invention is to provide a safe (non-toxic) treatment. Another objective is to provide an effective treatment that can delay, slow down or prevent the development of the disease, and possibly improve the patient's phenotype which can be monitored at the clinical level as indicated below.

[0178] In a subject, the composition according to the invention can be used:

[0179] - to improve mitochondrial function, particularly mitochondrial respiration;

[0180] - to improve growth;

[0181] - to improve muscle function;

[0182] - to improve vision and / or hearing;

[0183] - to improve respiratory function;

[0184] - to improve heart, liver or kidney functions;

[0185] - to improve brain functions;

[0186] - to improve digestive function; and / or

[0187] - to prolong survival, more generally to improve the quality and expectancy of life.

[0188] In one aspect, the invention provides 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 a composition as described above.

[0189] Advantageously, said improvements are observed up to 1 month after the start of treatment, or 3 months or 6 months or 9 months, more advantageously up to 1 year after the start of treatment, 2 years, 5 years, 10 years, or even throughout the subject's life.

[0190] In one embodiment, said improvements result in a reduction in the severity and / or frequency of symptoms and / or a delayed onset.

[0191] An improvement can be assessed based on methods known in the art, for example in the case of MELAS:

[0192] - assessment of lactate levels, particularly cerebral ventricular lactate, measured for example by magnetic resonance spectroscopy (MRS);

[0193] - assessment of quality and / or life expectancy using clinical scales, for example the NMDAS (Newcastle Mitochondrial Disease Scale for Adults) score or the SF-36 (Short Form Health Survey) score;

[0194] - assessment of brain changes, for example using magnetic resonance imaging (MRI);

[0195] - assessment of changes in muscle activity using physical tests such as the six-minute walk test; - assessment of changes in venous lactate and GDF 15 concentration;

[0196] - assessment of changes in mtDNA heteroplasmy in urine and blood.

[0197] The appropriate parameters for a given case can be adapted depending on the disease.

[0198] Thus, the claimed treatment makes it possible to improve the clinical condition and the various parameters disclosed above compared to an untreated subject.

[0199] The practice of the present invention employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, known to those skilled in the art. These techniques are explained in detail in the literature, in particular in “Molecular Cloning: A Laboratory Manual”, fourth edition (Sambrook, 2012); “Oligonucleotide Synthesis” (Gait, 1984); “Culture of Animal Cells” (Freshney, 2010); “Methods in Enzymology” “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); “Current Protocols in Immunology” (Coligan, 2002).Techniques particularly useful for particular embodiments will be discussed in the following sections.

[0200] The disclosures of each patent, patent application and publication cited herein are incorporated by reference in their entirety.

[0201] Without further description, it is believed that one skilled in the art can, using the following description and illustrative examples, make and use the compounds of the present invention and practice the claimed methods.

[0202] EXAMPLES OF ACHIEVEMENT

[0203] The invention and the advantages resulting therefrom will become more apparent from the following exemplary embodiments, supported by the appended figures. However, these are not intended to be limiting. FIGURE KEYS

[0204] Figure 1:

[0205] A / Effect of ebselen (EBS) on respiratory growth of several yeast models of mitochondrial diseases on non-fermentable solid medium, detected by the halo test

[0206] C / Effect (detected by the halo test) of ebselen oxide on the respiratory growth of the mutant yeast strain tazl on non-fermentable solid medium, in comparison with ebselen (B) and DMSO (D)

[0207] E / Effect of different analogues of ebselen (EBS) and ethaselen on respiratory growth of yeast strains tazl, find, cox2 and A29G.

[0208] Figure 2: Effect of ebselen (EBS) on the mutant yeast strain tazl

[0209] A / Respiratory growth on liquid medium

[0210] B / Accumulation of Cox2 protein

[0211] Figure 3: Effect of ebselen (EBS) on the syml mutant yeast strain

[0212] A / Respiratory growth on liquid medium

[0213] B / Cellular respiration

[0214] C / Accumulation of Cox2 protein

[0215] Figure 4: Effect of EBS on the thermosensitive growth of a Podospora anserina model carrying a mutation in mitochondrial complex I (Pa nuo-51 A357V ) A / Effect on the growth halo

[0216] B / Dose-dependent effect on growth

[0217] Figure 5: Effect of EBS on proliferation and viability of TAZ patient cells A / growth of TAZ patient fibroblasts in the presence or absence of EBS (12.15 nM)

[0218] B / 96-hour respiratory growth of TAZ patient fibroblasts as a function of EBS concentration (from 150 pM to 234.9 pM), compared to the DMSO negative control. C / 96-hour respiratory growth of TAZ patient fibroblasts as a function of ethaselen concentration (from 150 pM to 26.1 pM), compared to the DMSO negative control.

[0219] D / Mitochondrial ATP level in fibroblasts from TAZ patients, incubated in the presence of Antimycin A (100 pM), Rotenone (0.5 pM) or oligomycin A (3 pM), and EBS, at a concentration of 12.15 nM.

[0220] E / respiratory growth at 120 hours of myoblasts from a TAZ patient as a function of EBS concentration (from 50 pM to 234.9 pM).

[0221] Figure 6: Effect of EBS on the proliferation and viability of a HeLa cell line, in which the FMC1 gene has been deleted, in Ig / L glucose (A) or galactose (B) medium. OD (AU): optical density in arbitrary units. Figure 7: Effect of EBS on mitochondrial respiration of cells from patients deficient for complex I (CI) of the mitochondrial respiratory chain, carrying mutations affecting the nuclear gene NDUFV1 of the complex

[0222] A / Oxygen consumption rate (OCR)

[0223] B / basal breathing (routine)

[0224] C / F-linked respiration ATP synthase (RO)

[0225] D / maximum breathing

[0226] Figure 8: Synergistic effects between ebselen (EBS), disulfiram (DSF) and alverine (ALV)

[0227] A / in a taz mutant yeast model

[0228] B / in a Pa nuo-51 mutant A357V : growth rate in the presence of EBS and ALV alone or in combination, compared to the negative control DMSO (VEH)

[0229] Figure 9: Effect of EBS intake on the weight of KO mice lacking the TAZ gene (mouse model of Barth syndrome)

[0230] EXAMPLES 1 TO 3: SACCHAROMYCES CEREVISIAE MODEL

[0231] Each of the Saccharomyces cerevisiae yeast strains used in these examples contains different specific mutations, modeling human mutations that cause mitochondrial diseases. To varying degrees, all of these mutant yeast strains exhibit a growth defect when grown on a respiratory medium such as ethanol or glycerol at 28°C or 36°C (depending on the strain).

[0232] Mutated yeast strains:

[0233] - fmcl: MC6 MATa ade2- 1 his3 11.15 trpl 1 leu2- -3.112 ura3 1 fmcl::HIS3

[0234] [Ai ER OR] (Schwimmer, C. et al. (2005) J Biol Chem, 280, 30751-30759) for the first screening

[0235] - !MBA MATa ade2-l his3-ll,15 trpl-1 leu2-3,112 ura3-l CAN1 arg8::HIS3 p+ atp6- L173R or RKY20 MATa ade2-l his3-ll,15 trpl-1 leu2-3,112 ura3-l CAN1 arg8::HIS3 p+ atp6-L173P (Rak, M. et al. (2007) J Biol Chem, 282, 34039-34047) for the second screening

[0236] - tazlA: this strain was constructed by replacing the open reading frame of TAZ1 with that of TRP1 in strain W303-1A (MATa ade2-l ura3-l hisSll, 15 trpl-1 leu2-3,112 canl-100) (from Taffin de Tilques, M. et al. (2017) Dis Model Mech, 10, 439-450)

[0237] - shyl-G137R-, this mutant was constructed in the CW252 strain containing the W303 nuclear background and an intronless mitochondrial genome. This gene is a homologue of the human SURF1 gene

[0238] - symlA: this yeast strain was constructed by replacing the open reading frame of SYM1 with that of kanMXô in the W303-1 A strain MATa ade2-l ura3-l his311, 15 trpl-1 leu2-3,112 canl-100). This gene is a homologue of the human MPV17 gene

[0239] - cox2: MATa Iys2 leu-2-3,112 ura.3-52 his3AHindIII arg8::hisG cox2-22 (Bonnefoy, N. et al. (2001) Mol Cell Biol, 21, 2359-2372.

[0240] - A29G: The yeast strain used in these experiments includes tRNA-Leu A29G with MA genetic background Ta, his3 11, ade2 1, leu2 3, 112, ura3 1, trpl-D2, canl- 100, syn .

[0241] Associated mitochondrial diseases:

[0242] NARP syndrome

[0243] NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome induces a variable combination of developmental delay, retinitis pigmentosa, dementia, seizures, ataxia, proximal neurogenic muscle weakness, and sensory neuropathy. NARP is caused by various mutations in the mitochondrial ATP6 gene, which encodes the α-subunit of ATPase (complex V of the OXPHOS system). Mutations are often heteroplasmic (coexistence of mutated and wild-type (WT) mitochondrial DNA (mtDNA) in the same cells). Depending on the type of mutation and the percentage of mutated mtDNA (degree of heteroplasmy), the clinical consequences are more or less severe. ATP6 m.8993T>C / G mutations are among the most common in NARP patients and result in severe forms of NARP syndrome.FMC1 is a nuclear gene that encodes a protein required at high temperature (35-37°C) for the assembly of the F1 sector of ATP synthase, thus mimicking the heteroplasmy observed in NARP patients. Indeed, when cultured at a restrictive temperature (35-37°C), mitochondria of the fmclA mutant contain significantly fewer assembled ATP synthase complexes than a wild-type (WT) strain, but those that assemble are fully functional. This heterogeneity is also found in patients with reduced levels of ATP synthase due to heteroplasmic ATP6 mutations. Therefore, the fmclA mutant constitutes a suitable model of these disorders (Lefebvre-Legendre, L. et al. (2001) J Biol Chem, 276, 6789-6796). Various yeast NARP strains, carrying homoplasmic mutations equivalent to the T8993G and T8993C mutations (Rak, M. et al., J Biol Chem. 2007 282(47):34039-47), were also tested.

[0244] TAZ

[0245] The TAZ gene encodes tafazzin, a mitochondrial transacylase that catalyzes the remodeling of immature cardiolipin into its mature composition containing a predominance of tetralinoleoyl moieties. Mutations in TAZ result in Barth syndrome, an X-linked disorder classically characterized by dilated cardiomyopathy (DCM) with endocardial fibroelastosis (EFE), predominantly proximal skeletal myopathy, growth retardation, neutropenia, and organic aciduria, including excess 3-methylglutaconic acid (Barth, PG et al. (1996) J Inherit Metab Dis, 19, 157-160).

[0246] COX2 AND SURF1

[0247] The COX2 and SURF1 genes (SHY1 gene in yeast) encode a subunit and an assembly factor of mitochondrial complex IV, respectively. Mutations are associated with Leigh syndrome, a severe, progressive neurodegenerative disorder that manifests in the first months or years of life and can lead to early death. Affected individuals typically present with global developmental delay or developmental regression, hypotonia, ataxia, dystonia, and ophthalmologic abnormalities, such as nystagmus or optic atrophy (Barrientos, A. et al. (2002) EMBO J, 21, 43-52).

[0248] MPV17

[0249] MPV17 (SYM1 gene in yeast) encodes a mitochondrial inner membrane protein of unknown function. QMPV17 mutations result in:

[0250] - Mitochondrial DNA depletion syndrome-6, an autosomal recessive disease characterized by infantile onset of progressive liver failure, often leading to death within the first year of life. Those who survive develop progressive neurological impairment, including ataxia, hypotonia, dystonia, and psychomotor regression (Spinazzola, A. et al. (2006) Nat Genet, 38, 570-575).

[0251] - Navajo-type neuropathy: Manifestations include mainly severe sensorimotor peripheral neuropathy with hepatic and cerebral involvement such as leukoencephalopathy and corneal ulcerations. (Karadimas, CL et al. (2006) Am J Hum Genet, 79, 544-548).

[0252] - MELAS syndrome: The A30(29)G mutation in yeast A29G mimics the human m.3260A>G mutation of the tRNALeu (UUR) gene responsible for MELAS syndrome.

[0253] EXAMPLE 1: Effect of ebselen (EBS) and its derivatives on mutant yeast strains grown on a non-fermentable (respiratory) medium

[0254] MATERIALS AND METHODS

[0255] As previously described (WO2020 / 254632), the different mutant yeast strains were plated on solid agar-based respiratory media (containing glycerol or ethanol as the sole carbon source) and exposed to filters on which the test compound, namely ebselen (EBS), was deposited. The plates were then incubated at the indicated temperature (which can be 28°C or 36°C depending on the strain used).

[0256] Specifically, 0.125 OD of exponentially growing mutant cells were homogeneously plated with sterile glass beads on a square Petri dish (12 cm x 12 cm) containing solid respiratory medium (YPA Ethanol, 1% Yeast Extract, 0.5% Bacto Peptone, 40 mg / L Adenine, 2% Ethanol for syml and tazl or YPA Glycerol 2% for cox2, find, atp6, shyl and A29G). Small sterile filters were then placed on the agar surface and increasing concentrations of the test compounds were added to the filters at the indicated amounts (30 and 90 nmol). On each dish, DMSO (buffer) is used as a negative control (upper left filter). The dishes were then incubated at 28°C (for shyl or 36°C (for cox2, syml, find, atp6 tazl and A29G) for 4 to 5 days, then photographed.The improvement in growth was assessed after several days of incubation by the appearance of a halo of improved growth around the filter where the active ingredient was deposited.

[0257] RESULTS

[0258] As shown in Figure 1A, EBS is active on all mutant yeast strains tested, in a dose-dependent manner.

[0259] For the tazl mutant, Figure 1C shows that ebselen oxide (100 nmol) has comparable activity to ebselen (100 nmol) (Figure 1B), whereas no halo is observed in the presence of DMSO (Figure 1D).

[0260] More generally, different EBS and ethaselen analogues were evaluated on different mutants (tazl, find, cox2 and A29G1). As revealed in Figure 1E, all tested analogues are active on the tested strains.

[0261] EXAMPLE 2: Effect of ebselen (EBS) on the mutant yeast strain tazl

[0262] MATERIALS AND METHODS

[0263] Respiratory growth in liquid culture:

[0264] To determine the optimal EBS concentration leading to the suppression of the respiratory growth defect of the mutant yeast strain, exponentially growing cells were inoculated into fresh non-fermentable YPA Ethanol media supplemented, or not, with increasing concentrations of EBS. Cell growth in the liquid respiratory medium was monitored with the bioscreen system over a period of 80 h during which cell densities (OD600nm) were taken every 20 minutes. Cellular respiration:

[0265] Respiratory intensity corresponds to the amount of oxygen consumed in relation to time and cell number. It reflects the mitochondrial oxidative metabolism of cells. Oxygen consumption is measured using an OROBOROS oxygraphic system. Cells are cultured for 7-8 generations at 28°C for 24 or 48 h in YPA Ethanol medium supplemented with DMSO or EBS (IpM). 10 7 cells are introduced into the oxygraph. CL consumption is recorded with or without CCCP (maximal oxygen consumption rate). The CL consumption rate is calculated based on the linear part of O2 consumption.

[0266] Accumulation of Cox2 protein:

[0267] Total protein extracts from the mutant yeast strain were analyzed by SDS-PAGE (50 pg per well) using antibodies against the indicated proteins. The gels shown are representative of at least 3 experiments. Protein levels were quantified using ImageJ software. Cox2 levels are normalized to Adel3p and expressed relative to the wild-type (WT) strain.

[0268] RESULTS

[0269] As shown in Figure 2 A, EBS is toxic from 27 pM and active between 1 nM and 3 pM, with an optimal concentration of 1 pM on the tazl mutant strain.

[0270] Furthermore, and as shown in Figure 2B, EBS IpM increases the accumulation of Cox2 protein, a subunit of complex IV of the mitochondrial respiratory chain, in the tazl mutant strain.

[0271] EXAMPLE 3: Effect of ebselen (EBS) on the mutant yeast strain syml

[0272] The experiments described in Example 2 were carried out on the mutant yeast strain syml.

[0273] The results are shown in Figure 3.

[0274] Data are the means ± SEM of at least three independent experiments. The significance of variations between samples and controls was estimated using the multifactorial Anova: Tukey test.

[0275] As with the tazl mutant strain, at the optimal EBS concentration (1 pM), cellular respiration and Cox2 levels are increased compared to untreated cells. EXAMPLE 4: PODOSPORA ANSERINA model

[0276] The yeast Saccharomyces cerevisiae, used in Examples 1 to 3, does not possess complex I of the mitochondrial respiratory chain. EBS was therefore tested on the strictly aerobic filamentous fungus Podospora anserina.

[0277] The strain used in the following examples contains a specific mutation modeling a human mutation in the NDUF V 1 subunit of complex I resulting in mitochondrial disease. This strain exhibits a growth defect at a non-permissive temperature above 31.5°C.

[0278] Mutated P. anserina strain: nuo-51 A357V : The A357V mutation was introduced into the nuo-51 gene of the wild-type S strain in association with a nourseothricin resistance cassette (NatR). In order to best mimic the human disease NDUFV1 A341 V, the NDI-1 and AOX genes were inactivated (El-Khoury et al., (2008) Curr Genet. 53:249-58)

[0279] Strain genotype: S, mat~, nuo-51 A35TV t \ndi-l, \aox, Naf, Hygrc

[0280] MATERIALS AND METHODS

[0281] The temperature-sensitive mutant Pa nuo-51 A357Vwas spread evenly with sterile glass beads on a square Petri dish containing solid minimal medium (M2). Small sterile filters were then placed on the agar surface and 20 nmol of EBS was added. DMSO (the vehicle of the compound) was used as a negative control. The plate was then incubated at the non-permissive temperature of 33°C for 4-5 days and then photographed.

[0282] The mutant Pa nuo-51 A357V was then cultured at 32°C where its growth was reduced compared to the wild-type strain (WT). The effect of EBS on the growth rate of the mutant was determined on minimal medium supplemented or not with increasing concentrations of EBS and estimated in centimeters of growth per day (cm / day).

[0283] RESULTS

[0284] Figure 4 A shows a positive effect of EBS on the growth of the mutant. Specifically, between 0.003 and 0.03 pM, EBS shows a significant improvement in the growth of the Pa nuo-51 mutant. A357V (Figure 4B). EXAMPLE 5: Human cell model

[0285] MATERIALS AND METHODS

[0286] Statistical analyses were performed using the two-way mixed ANOVA statistical test and applying the Bonferroni multiple testing correction, or with the graphpad prism 9.5.1 software.

[0287] The number of stars indicates the p-value < 0.05 (*), < 0.001 (**), < 0.0001 (***), < 0.0001 (****).

[0288] Fibroblasts and myoblasts from TAZ patients:

[0289] EBS was tested for its effect on respiratory growth of TAZ patient fibroblasts, namely fibroblasts from eight different patients with mutations in the TAZ gene (with Barth syndrome). Data represent the percentage growth relative to T0.

[0290] The proliferation rate was measured in low glucose medium (Ig / L; to force cells to use the OXPHOS system rather than glycolysis) in 384-well microtiter plates (1200 cells / well at T0) using the Sulforhodamine B (SRB) staining protocol at 24, 48, 72, or 96 hours after cell placement with or without 12.15 nM EBS.

[0291] The proliferation rate was then measured under the same experimental conditions but with varying EBS concentrations: Cells were analyzed 96 hours after treatment with increasing doses of EBS (from 150 pM to 234.9 pM). Data are the average of 8 patients or 3 controls in triplicate, in 4 independent experiments.

[0292] The proliferation rate was also measured under the same experimental conditions but with varying ethaselen concentrations: Cells were analyzed 96 hours after treatment with increasing doses of ethaselen (from 150 pM to 26.1 pM). Bars represent the mean of 4 replicates from 2 patients in 3 independent experiments.

[0293] Mitochondrial ATP levels in TAZ patient fibroblasts were assessed using the Promega CellTiter-Glo kit. Cells in 96-well plates (5000 / well) were treated or not with 12.15 nM EBS for 96 hours and then incubated in the presence of Antimycin A (100 pM), Rotenone (0.5 pM), or Oligomycin A (3 pM).

[0294] EBS was also tested for its ability to restore cell growth in a patient myoblast line carrying mutations in the TAZ gene. The proliferation rate was measured in low-glucose medium in 384-well microtiter plates (1200 cells / well at T0) using the Sulforhodamine B (SRB) staining protocol at 48, 72, 96, and 120 hours after cell plating. The graph represents the growth rate at 120 hours, corresponding to the average of 3 independent experiments.

[0295] FMC1 KO (“knockout”) cell line:

[0296] The HeLa KO FMC1 cell line, mimicking human diseases related to ATP synthase deficiency (NARP syndromes, MILS), was generated by the CRISPR-Cas9 method. The HeLa KO FMC1 cell line is cultured in DMEM medium containing 4.5 g / L glucose and 110 mg / L pyruvate and 50 mg / L uridine.

[0297] The proliferation rate of this HeLa cell line in which the FMC1 gene was deleted was measured in a medium containing either low glucose (1 g / L), or galactose in 384-well microplates (4500 cells / cm 2 at T0) using the Sulforhodamine B (SRB) staining protocol 96 hours after cell seeding and treatment. Bars represent the mean of 8 replicates in 3 independent experiments. Data represent the percentage growth relative to T0.

[0298] CI (complex I) KO (“knockout”) cell line:

[0299] EBS was also tested for its ability to restore mitochondrial respiration in fibroblast cell lines from patients carrying mutations in the nuclear mitochondrial complex I gene NDUFV1. Human NDUFV1 cell lines mimicking human diseases related to mitochondrial complex I deficiency (Leigh syndrome) carry genetic mutations affecting the NDUFV1 gene. These cell lines are cultured in DMEM culture medium containing 1.0 g / L glucose and 1 / 1M sodium pyruvate and 50 pg / mL uridine (Sigma Aldrich, Lyon, France).

[0300] Respiration was measured using a Seahorse XF96 extracellular flux metabolic analyzer in 96-well plates according to the manufacturer's protocol (Agilent Technologies). Deficient cells were exposed for 48 h to different concentrations with a dose-response curve combining a concentration range from 25 nM to 10 pM. In (A) is the oxygen consumption rate, in (B) basal respiration, in (C) ATP synthase-mediated respiration, and in (D) maximal respiration. Stars represent the average of 4 independent replicates.

[0301] Primary fibroblasts in the exponential growth phase were detached by the addition of 2 ml of 0.05% trypsin. These fibroblasts were used to seed XF96 plates (30 x 10 3cells / well) kept in culture for 4 hours. Cells were then incubated in bicarbonate-free DMEM (Sigma-Aldrich) supplemented with 5.5 mM glucose, 2 mM L-glutamine, in a CO2-free incubator for 1 h. Oxygen consumption rate (OCR) was recorded under basal conditions to assess mitochondrial respiratory activity and then cells were treated sequentially with 4 μg / mL oligomycin and 1.5 μM carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) (Sigma-Aldrich). Basal OCR, ATP-dependent respiration, maximal respiratory capacity and proton leak respiration were determined. Non-mitochondrial respiration (OCR after treatment with 2 μg / mL antimycin A) was subtracted from all OCR measurements.ATP-related respiration (OR) was estimated from the difference between basal and oligomycin-inhibited respiration rates, and proton leak respiration was obtained by subtracting nonmitochondrial respiration from the OCR measured after oligomycin treatment. Maximal respiratory capacity was determined as the respiration rate in the presence of the uncoupler FCCP. Results were normalized to cell number.

[0302] RESULTS

[0303] Figure 5A shows that EBS at 12.15 nM enhances respiratory growth of fibroblasts from TAZ patients. At this concentration, EBS is effective from 24 hours and the improvement in cell proliferation is more pronounced at the other three sampling times (48, 72 and 96 hours).

[0304] Figure 5B shows that at 96 hours, EBS enhances respiratory growth of TAZ patient fibroblasts across a wide range of concentrations: EBS has beneficial effects from 1.35 nM to 8.7 pM with toxicity beyond 234.9 pM, compared to the negative control DMSO.

[0305] Figure 5C shows that at 96 hours, ethasalen also enhances proliferation of cells from patients with Barth syndrome across a wide concentration range from approximately 1 nM to 3 pM.

[0306] Figure 5D shows that EBS, at a concentration of 12.15 nM, increases mitochondrial ATP production in fibroblasts from TAZ patients.

[0307] The effect of EBS on restoring growth of a patient myoblast line carrying mutations in the TAZ gene is illustrated in Figure 5E: it shows that EBS from 450 pM to 8.7 pM has beneficial effects compared to the negative control DMSO, with toxicity above 78.3 pM.

[0308] Figure 6 shows that ebselen enhances proliferation of model cells of ATP synthase deficiency (NARP, MILS syndromes) in a dose-dependent manner and across a wide concentration range (pM to pM). Figure 7 reveals that ebselen enhances cellular respiration of model cells of complex I-related mitochondrial pathologies from 25 nM to 100 nM and becomes toxic above 10 pM.

[0309] EXAMPLE 6: Synergies between different compounds

[0310] Disulfiram (DSF) and alverine (ALV) were reported in WO2020 / 254632 and W02022 / 018297, respectively, as useful in the management of pathologies associated with mitochondrial dysfunction.

[0311] Therefore, a possible synergistic effect between ebselen (EBS) and these compounds was tested in different models:

[0312] - in a taz mutant yeast model

[0313] - in a complex I mutant of Podospora anserina (Pa nuo-51 S5 V )

[0314] - in a taz mutant mammalian cell model.

[0315] MATERIALS AND METHODS

[0316] Taz yeast:

[0317] As in Figure 1, the mutant yeast strain tazl was plated on a solid respiratory medium containing ethanol as the carbon source. The strain was then exposed to filters on which were deposited either disulfiram (DSF 5 nmol), ebselen (EBS 30 nmol) or alverine (ALV 30 nmol), alone or in combination of two or three. The plates were then incubated at 36°C and scanned after 5 days of incubation. DMSO (solvent) is used as a negative control (upper left filter).

[0318] Pa nuo-51 A357V :

[0319] Growth rates at 32°C, estimated in centimeters of growth per day, were determined in minimal medium supplemented with EBS (0.003 pM, 0.01 pM or 0.03 pM), ALV (1 pM) or a combination of concentrations of both, compared with the same concentration of DMSO (VEH) per plate.

[0320] Mammalian cell taz:

[0321] EBS, ALV, and DSF were tested for their synergistic effect on the respiratory growth of myoblasts from TAZ patients carrying mutations in the TAZ gene. The proliferation rate was measured in low-glucose medium (to encourage cells to use the OXPHOS system rather than glycolysis) in 384-well microtiter plates (1200 cells / well at T0) using the Sulforhodamine B (SRB) staining protocol at 120 hours after seeding the cells with or without the different compounds alone or in combination at the indicated concentrations. RESULTS

[0322] Figure 8A shows that the combination of the 3 molecules EBS, ALV and DSF increases the growth halo around the filter where they were deposited in association, suggesting a synergistic effect of these 3 drug candidates on the respiratory growth of the tafazzin-deficient yeast model.

[0323] Figure 8B shows that the combination 1 pM ALV + 0.003 pM EBS significantly increases the growth of the Pa nuo-51 mutant A357V , compared to IpM ALV alone, suggesting a synergistic effect on the growth of this complex I mutant.

[0324] EXAMPLE 7: Mouse model of Barth syndrome

[0325] MATERIALS AND METHODS

[0326] Mice lacking the TAZ gene (KO, Barth syndrome model) or wild-type (WT) were gavaged twice daily with 10 mg / kg of Ebselen from the age of 51 days. Weight was monitored regularly during 1 month of treatment.

[0327] RESULTS

[0328] As shown in Figure 9, EBS-treated KO mice appeared to gain more weight than solvent-treated mice.

Claims

CLAIMS 1. Pharmaceutical composition comprising a compound having a group of formula: for use in the treatment of a disease associated with mitochondrial dysfunction.

2. Composition for use according to claim 1, wherein the compound is of formula: in which E is O, S, SO, SO2, Se or SeO each of the phenyl rings A and B is optionally substituted by one or more substituents, wherein each substituent is independently selected from: - a halogen, which is preferably selected from F, Cl and Br - an alcohol - an amine - a nitro - a C1-C4 alkyl, such as a C1-C2 alkyl or a Cl alkyl, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and - a C1-C4 alkoxy, such as a C1-C2 alkoxy or a Cl alkoxy, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br.

3. Composition for use according to claim 2, wherein the compound is ethaselen or one of its derivatives, for example MAD423.

4. Composition for use according to claim 1, wherein the compound is of formula: in which E is O, S, SO, SO2, Se or SeO each of the phenyl rings A and B is optionally substituted by one or more substituents, wherein each substituent is independently selected from: - a halogen, which is preferably selected from F, Cl and Br - an alcohol - an amine - a nitro - a C1-C4 alkyl, such as a C1-C2 alkyl or a Cl alkyl, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and - a C1-C4 alkoxy, such as a C1-C2 alkoxy or a Cl alkoxy, optionally substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br.

5. Composition for use according to claim 4, wherein the compound is: - 2-phenyl-1,2-benzisoselenazol-3(2H)-one or ebselen of formula: OR - l-oxyde-2-phenyl-l,2-benzisoselenazol-3(2H)-one or ebselen oxide or ebselen selenoxide of formula:

6. Composition for use according to any one of claims 1 to 5, according to which the disease is a disease of the mitochondrial respiratory chain, advantageously associated with a deficiency of complex I.

7. Composition for use according to any one of claims 1 to 6, wherein the disease is a genetic disease.

8. Composition for its use according to claim 7, according to which the genetic disease comprises at least one mutation in at least one of the following genes: MTND1 (or ND1 MTND2 (or ND2 MTND3 (or ND3 MTND4 (or ND4), MTND5 (or ND5), MTND6 (or ND6), MTND4L (or ND4E), 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, NDUFAF1, NDUFAF2, NDUFAF3, NDUFAF4, NDUFAF5, NDUFAF6, NDUFAF7, NDUFAF8, NUBPL, ACAD9, TMEM70, TMEM126B, FOXRED1, ECSIT, AIF, TIMMDC1.

9. Composition for use according to claim 7, wherein the genetic disease comprises at least one mutation in at least one of the following genes: MTTL1, ATP6, FMC1, 1AZ, C0X2, SURF1, POI.G, MPV17, OPA1, COA6, ND6 and BCS1L, advantageously A TP6, FMC1, PAZ, C0X2, SURF1 or MPV17.

10. Composition for use according to any one of claims 1 to 9, according to which the disease is chosen from the group consisting of: MELAS syndrome, maternally inherited myopathy and cardiomyopathy, NARP or MILS syndrome, Leigh syndrome, Leber hereditary optic neuropathy (LHON), Barth syndrome, mitochondrial DNA depletion syndrome including 4A (Alpers type) and 4B (MNGIE type), recessive mitochondrial ataxia syndrome, ataxic sensory neuropathy, dysarthria and ophthalmoplegia, spinocerebellar ataxia with epilepsy, progressive external ophthalmoplegia, mitochondrial DNA depletion syndrome-6, Navajo-like neuropathy, Behr syndrome, mitochondrial DNA depletion syndrome-14, infantile cardioencephalomyopathy due to cytochrome c oxidase deficiency (COA6 mutations), mitochondrial complex III nuclear deficiency type 1, GRACILE syndrome, and Bjomstad syndrome.

11. Composition for use according to claim 10, wherein the disease is selected from the group consisting of: Leigh syndrome, Leber hereditary optic neuropathy (LHON), MELAS syndrome, NARP or MILS syndrome, Barth syndrome, mitochondrial DNA depletion syndrome and Navajo type neuropathy.

12. Composition for use according to any one of claims 1 to 7, wherein the disease is chosen from the group consisting of: - 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 diseases, such as 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 illnesses, such as autism spectrum disorders, schizophrenia, bipolar and mood disorders; - gastrointestinal disorders; - fatigue diseases, such as chronic fatigue syndrome and Gulf War illness; musculoskeletal diseases, such as fibromyalgia and skeletal muscle hypertrophy / atrophy; - muscular dystrophies, - cancer; and - chronic infections.

13. Composition for use according to any one of the preceding claims, wherein the composition comprises another compound for treating the same disease, advantageously alverine and / or disulfiram.

14. Composition for use according to any one of the preceding claims, wherein the composition is administered orally.

15. Composition for use according to any one of the preceding claims, wherein the composition is in a solid form such as a tablet, even more advantageously comprising 200 mg of ebselen.

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