Use of ebseselenine or one of its derivatives for treatment of mitochondrial conditions or dysfunction

Abstract

The present invention relates to the use of ebseoline or one of its derivatives for the treatment of a mitochondrial condition or dysfunction, in particular a mitochondrial condition or dysfunction associated with genetic complex I deficiency.

Description

Technical Field

[0001] The present invention aims to provide new pharmacological tools for the treatment of mitochondrial diseases or dysfunctions, in particular mitochondrial diseases or dysfunctions associated with mitochondrial complex I defects. Background Art

[0002] Mitochondrial diseases are common metabolic conditions characterized by high clinical and genetic heterogeneity, which manifests itself in the fact that mitochondria cannot successfully produce enough energy to allow the organism to function correctly. Mitochondrial diseases may be present at birth but may also occur at any age. At the genetic level, mitochondria have the particularity that they have their own DNA (or mitochondrial DNA), but most mitochondrial proteins depend on the nuclear genome. Therefore, genetic abnormalities carried by the mitochondrial genome as well as the nuclear genome cause mitochondrial diseases, which reflects very high clinical and genetic heterogeneity. It is estimated that one in 4300 people is affected by mitochondrial diseases (Gorman et al., 2015, Ann Neurol. 77 (5): 753-9).

[0003] Mitochondrial diseases can affect almost every part of the body, especially cells of the brain, nerves, muscles, kidneys, heart, liver, eyes, ears, or pancreas. Mitochondrial disease symptoms depend on the organ involved, preferentially affecting high-energy-consuming tissues like muscles, brain, and heart. A patient's symptoms may be moderate to severe for one or more organs.

[0004] Symptoms of mitochondrial disease may include:

[0005] - Poor growth

[0006] - Muscle weakness, myalgia, poor muscle tone, inability to exert force

[0007] - Visual and / or hearing impairment

[0008] -Learning difficulties, developmental delay, intellectual disability

[0009] - Autism or autistic traits

[0010] - Heart dysfunction, arrhythmia, or cardiac conduction abnormalities

[0011] - Liver or kidney disease

[0012] - Gastrointestinal disturbances, difficulty swallowing, diarrhea or constipation, unexplained vomiting, cramps, gastrointestinal reflux

[0013] -diabetes

[0014] - Increased risk of infection

[0015] - Neurological disorders, epileptic seizures, migraines, cerebrovascular accidents (strokes)

[0016] - Movement disorders

[0017] - Thyroid and / or adrenal dysfunction

[0018] - Breathing disorders

[0019] - Lactic acidosis, which is the accumulation of lactic acid in the blood or urine

[0020] -dementia.

[0021] Mitochondrial dysfunction can also occur when mitochondria are not functioning properly, which may be due to another disease or another illness. Many conditions can cause secondary mitochondrial dysfunction, especially Alzheimer's or Parkinson's disease, muscular dystrophy, Lou Gehrig's disease, diabetes, and cancer. People with secondary mitochondrial dysfunction do not have the primary mitochondrial disease of genetic origin, but have similar symptoms. In addition, certain drugs can target or damage mitochondria.

[0022] Mitochondrial complex I deficiency is the most common defect observed in more than 30% of mitochondrial diseases. Among them, the two most common clinical phenotypes associated with complex I deficiency are the often fatal Leigh syndrome or milder phenotypes such as Leber hereditary optic neuropathy (LHOL). MELAS syndrome is also considered a common mitochondrial disease due to mutations in the mitochondrial genome and is associated with a severe reduction in mitochondrial complex I activity.

[0023] Complex I consists of at least 44 subunits, of which 7, 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 very variable. Among the subunits of complex I, mutations targeting the gene NDUFV1 have been shown to be the cause of severe neurological phenotypes (Schuelke et al., 1999, Nat Genet. 21 (3): 260-61). Mutations affecting subunit NDUFS8 have been associated with Leigh syndrome (Procaccio et al., 2004, Neurology 62: 1899), and mutations targeting the subunit ND3 encoded by mitochondrial DNA have been reported in Leigh syndrome and in LHOL (Sarzi et al., 2007 American Journal of Medical Genetics 143: 33-41; Wang et al., 2009, Neurogenetics. 10: 337-345). Furthermore, mutations affecting the subunit ND6 encoded by mitochondrial DNA have been reported in LHOL (Johns et al., 1992, Biochem Biophys Res Commun. 187(3): 1551-7).

[0024] Furthermore, complex I defects have been identified in secondary mitochondrial dysfunction associated with age-related neurodegenerative diseases such as Parkinson's disease.

[0025] Even though most mitochondrial diseases have a genetic origin, genetic therapy appears difficult to implement due to the genetic and clinical diversity and complexity of these diseases.

[0026] The goal of current treatment is to relieve symptoms and slow the progression of disease or disability using suggestions such as the following:

[0027] -Use vitamin therapy

[0028] -Save energy

[0029] - Slow down activities

[0030] - Keep the environment at room temperature

[0031] -Prevent exposure to complications (infection, stress)

[0032] -Maintain nutrition and proper hydration.

[0033] It should be noted that documents WO2020 / 254632 and WO2022 / 018297 recently reported the use of disulfiram and alverine, respectively, for the treatment of these diseases or defects.

[0034] However, there is still a need to find new therapeutic and pharmacological approaches to treat these types of dysfunctions or diseases based on etiological and non-symptomatic treatments.

[0035] Document WO 2014 / 150688 describes the use of molecules such as ebselen that are able to inhibit enzymes from the mitochondrial chain (1-C), in particular the enzyme MTHFD2, for the treatment of cancer.

[0036] JIA ZHI-QIANG et al. (Neurosc. Letters, 2018, vol. 678, pp 110-17) reported the beneficial effects of ebselen in the treatment of acute spinal cord injury, with neuroprotective effects and improved mitochondrial function.

[0037] The document ARAKAWA MOTOKI et al. (Cerebellum, 2007, 6 (4), pp 308-14) relates to neurodegenerative diseases such as Alzheimer's disease or Parkinson's disease, which are described as being associated with mitochondrial changes. The document focuses on the positive effects of NAC (N-acetylcysteine) on neurons against toxicity induced by 4-hydroxynonenal (HNE) and mentions the neuroprotective effect of ebselen and its possible use in combination with NAC.

[0038] The literature AZAD GAJENDRA et al. (Molecular Biology Reports, 2014, 41(8), pp4865-79) is a review focused on ebselen, which is described as a promising antioxidant drug. Table 2 lists all the conditions that can be improved by administering ebselen.

[0039] The document CAPPER MICHAEL et al. (Nature Communications, 2018, 9(1)) concerns the effect of ebselen on the enzyme superoxide dismutase 1 (SOD1), mutations of which may lead to conditions such as amyotrophic lateral sclerosis (ALS). Summary of the invention

[0040] The inventors have shown that ebselen, a drug currently in clinical trials for the treatment of hearing disorders such as Meniere's disease, and its derivatives such as ethasalen, are effective in the context of the treatment of diseases associated with mitochondrial dysfunction, in particular diseases of the mitochondrial respiratory chain, advantageously associated with complex I defects. This work therefore opens the way to the use of a new family of compounds in this context.

[0041] definition

[0042] The following definitions give the general meanings used within the framework of the present invention and have to be taken into account unless another definition is explicitly indicated.

[0043] The terms "about," "approximately," "in the order of," and "approximately" used in this document to specify measurable values ​​(such as amounts, times, etc.) should be understood to encompass variations of ±20% or ±10%, preferably ±5%, more preferably ±1%, and still more advantageously ±0.1% relative to the specified value.

[0044] Intervals / ranges: Throughout this disclosure, various aspects of the invention may be presented in the form of intervals of values ​​(ranges). It must be understood that describing values ​​in interval form is only for convenience and brevity, and should not be interpreted as limiting the scope of the invention. Therefore, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values ​​within this range. For example, a 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 numbers within the range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the extent of the range.

[0045] "Isolated" means extracted or taken from its environment or natural state. For example, an isolated nucleic acid or peptide is a nucleic acid or peptide extracted from the natural environment in which the nucleic acid or peptide is normally found, whether, for example, in a plant or a living animal. For example, a nucleic acid or peptide naturally present in a living animal is not an isolated nucleic acid or peptide within the meaning of the present invention, but the same nucleic acid or peptide itself, partially or completely separated from other components present in its natural environment, is "isolated" within the meaning of the present invention. An isolated nucleic acid or peptide may be present in a substantially purified form, or may be present in a non-natural environment (e.g., a host cell).

[0046] The term "abnormal," when used in the context of an organism, tissue, cell, or component thereof, refers to an organism, tissue, cell, or component thereof that differs from an organism, tissue, cell, or component thereof that exhibits a corresponding "normal" (expected) property by at least one observable or detectable property (e.g., age, treatment, day, etc.). A property that is 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", etc. are used interchangeably in this document and refer to any animal or cell thereof that can be subjected to the methods described in this document in vitro or in situ. In certain non-limiting embodiments, the patient, subject or individual is an animal, preferably a mammal, more advantageously a human, whether male or female. The animal may also be a mouse, rat, pig, dog or non-human primate (NHP), such as a macaque.

[0048] A "disease" or "condition" within the meaning of the present invention is a health state of an animal in which the animal's homeostasis is negatively affected and continues to deteriorate if the disease is not treated. On the other hand, a "disorder" or "malfunction" within the meaning of the present invention refers to a health state in which the animal is able to maintain its homeostasis, but in which the animal's health is not as good as 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 health over time.

[0049] A disease or "disorder" is "attenuated" ("reduced") or "improved" if the severity of the symptoms of the disease or disorder, the frequency with which the subject feels the symptoms, or both, is reduced. The definition also includes the disappearance 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 the symptoms of the disease or disorder, the frequency with which the subject feels the symptoms, or both, are eliminated.

[0050] In the context of the present invention, a "therapeutic" treatment is a treatment given to a subject with symptoms (signs) of a pathology with the aim of reducing or eliminating said symptoms. In the framework of the present invention, "treatment of a disease or disorder" refers to a reduction in the frequency or severity of at least one sign or symptom of the disease or disorder experienced by the subject. When a treatment is given to prevent the development, spread or aggravation of a disease, particularly if the subject does not present or has not yet presented symptoms of the disease and / or the disease has not yet been diagnosed for him, the treatment is said to be preventive.

[0051] As used herein, "treating a disease or disorder" refers to reducing the frequency or severity of at least one sign or symptom of the disease or disorder experienced by a subject. Disease or disorder may be used interchangeably in the context of treatment according to the present invention.

[0052] Within the meaning of the present invention, a "useful amount" or "effective amount" of a compound is an amount of the compound sufficient to provide a beneficial effect to a subject to which the compound is administered. The expression "therapeutically effective amount" refers to an amount that is sufficient or effective to prevent or treat (among other aspects, delay or prevent development, prevent progression, inhibit, reduce, diminish or reverse) a disease or disorder (including alleviating the symptoms of the disease or disorder).

[0053] The present invention relates to the use of the following for treating diseases associated with mitochondrial dysfunction or for preparing a medicament intended to treat diseases associated with mitochondrial dysfunction:

[0054] A compound having a moiety of the formula:

[0055]

[0056] Or a compound of the formula:

[0057]

[0058] or a compound of the formula

[0059]

[0060] in,

[0061] E is O, S, SO (S=O), SO2 (O=S=O), Se or SeO (Se=O),

[0062] The benzene rings A and B may each be substituted by one or more substituents.

[0063] wherein each substituent is independently selected from:

[0064] - halogen, which is preferably selected from F, Cl and Br,

[0065] -alcohol,

[0066] -amine,

[0067] -Nitro,

[0068] - C1-C4-alkyl, such as C1-C2-alkyl or C1-alkyl, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0069] - C1-C4-alkoxy, such as C1-C2-alkoxy or C1-alkoxy, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br,

[0070] Or a pharmaceutical composition containing the halogen.

[0071] In other aspects, the present invention relates to the following for use in treating diseases associated with mitochondrial dysfunction:

[0072] A compound having a moiety of the formula:

[0073]

[0074] Or a compound of the formula:

[0075]

[0076] or a compound of the formula

[0077]

[0078] in,

[0079] E is O, S, SO (S=O), SO2 (O=S=O), Se or SeO (Se=O),

[0080] The benzene rings A and B may each be substituted by one or more substituents.

[0081] wherein each substituent is independently selected from:

[0082] - halogen, which is preferably selected from F, Cl and Br,

[0083] -alcohol,

[0084] -amine,

[0085] -Nitro,

[0086] - C1-C4-alkyl, such as C1-C2-alkyl or C1-alkyl, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0087] - C1-C4-alkoxy, such as C1-C2-alkoxy or C1-alkoxy, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br,

[0088] Or a pharmaceutical composition containing the halogen.

[0089] According to another aspect, the present invention relates to a method for treating a disease associated with mitochondrial dysfunction, the method comprising administering to a subject the following for use in treating a disease associated with mitochondrial dysfunction:

[0090] A compound having a moiety of the formula:

[0091]

[0092] Or a compound of the formula:

[0093]

[0094] or a compound of the formula

[0095]

[0096] in,

[0097] E is O, S, SO (S=O), SO2 (O=S=O), Se or SeO (Se=O),

[0098] The benzene rings A and B may each be substituted by one or more substituents.

[0099] wherein each substituent is independently selected from:

[0100] - halogen, which is preferably selected from F, Cl and Br,

[0101] -alcohol,

[0102] -amine,

[0103] -Nitro,

[0104] - C1-C4-alkyl, such as C1-C2-alkyl or C1-alkyl, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0105] - C1-C4-alkoxy, such as C1-C2-alkoxy or C1-alkoxy, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br,

[0106] Or a pharmaceutical composition comprising said halogen.

[0107] The compounds implemented within the framework of the present invention may have an activity of inhibiting inositol monophosphatase (IMP enzyme), which activity can be tested as described by Singh et al. (Nat Commun. 2013; 4: 1332. doi: 10.1038 / ncomms2320). According to a specific embodiment, such a compound is not valproic acid or carbamazepine.

[0108] The compounds implemented within the framework of the present invention may have antioxidant activity which can 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).

[0109] According to a specific embodiment, such compound is not ascorbic acid.

[0110] According to a specific embodiment, the compound implemented within the framework of the present invention having a group of the formula is ethaselenol or one of its derivatives or analogs as defined below:

[0111]

[0112] Ethaneselenol or BBSKE (CAS No. 217798-39-5) has the following formula:

[0113]

[0114] Derivatives or analogs of ethaselenine are, for example, chlorinated derivatives MAD423 of the following formula:

[0115]

[0116] Therefore, compounds of interest within the framework of the present invention have the following formula:

[0117]

[0118] in,

[0119] E is O, S, SO, SO2, Se or SeO,

[0120] The benzene rings A and B may each be substituted by one or more substituents.

[0121] wherein each substituent is independently selected from:

[0122] - halogen, which is preferably selected from F, Cl and Br,

[0123] -alcohol,

[0124] -amine,

[0125] -Nitro,

[0126] - C1-C4-alkyl, such as C1-C2-alkyl or C1-alkyl, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0127] - C1-C4-alkoxy, such as C1-C2-alkoxy or C1-alkoxy, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br.

[0128] According to another embodiment, the compound implemented within the framework of the present invention is as described in document WO2012 / 107735, ie a compound of formula I or a pharmaceutically acceptable salt thereof, wherein formula I is:

[0129]

[0130] in,

[0131] E is O, S, SO, SO2, Se or SeO,

[0132] The benzene rings A and B may each be substituted by one or more substituents.

[0133] wherein each substituent is independently selected from:

[0134] - halogen, which is preferably selected from F, Cl and Br,

[0135] -alcohol,

[0136] -amine,

[0137] -Nitro,

[0138] - C1-C4-alkyl, such as C1-C2-alkyl or C1-alkyl, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br; and

[0139] - C1-C4-alkoxy, such as C1-C2-alkoxy or C1-alkoxy, which may be substituted by one or more halogen atoms, each of which is preferably selected from F, Cl and Br.

[0140] According to a preferred embodiment, such a compound is:

[0141] - 2-phenyl-1,2-benzisoselenazol-3(2H)-one or ebselen of the following formula:

[0142]

[0143] - 2-phenyl-1,2-benzisothiazol-3(2H)-one or ebselenthiolate (2-phenyl-2,3-dihydro-1,2-benzothiazol-3-one) of the following formula:

[0144]

[0145] - 1-oxide-2-phenyl-1,2-benzisoselenazol-3(2H)-one or ebselenoxide (ebselenoxide) or ebselenoselenoxide of the following formula:

[0146]

[0147] - Ebselen -OH para-1 of the following formula:

[0148]

[0149] - The ortho position of the ebselen-OH of the following formula:

[0150]

[0151] - Ebselen -OH para-2 of the following formula:

[0152]

[0153] -MAD281 (6-methoxy-2-phenylbenzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0154]

[0155] -MAD309 (6-hydroxy-2-phenylbenzo[d][1,2]selenazol-3(2H)-one 1-oxide) of the following formula:

[0156]

[0157] -MAD331 (6-chloro-2-phenylbenzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0158]

[0159] -MAD332 (6-fluoro-2-phenylbenzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0160]

[0161] -MAD349 (2-(2,6-dichlorophenylbenzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0162]

[0163] -MAD380 (2-(3-hydroxyphenylbenzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0164]

[0165] -MAD383 (6-chloro-2-(3-trifluoromethyl)phenyl)benzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0166]

[0167] -MAD384 (2-(3-nitrophenyl)benzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0168]

[0169] -MAD385 (2-(3-difluoromethoxy)phenyl)benzo[d][1,2]selenazol-3(2H)-one) of the following formula:

[0170]

[0171] -MAD413 (6-chloro-2-phenylbenzo[d][1,2]selenazol-3(2H)-one 1-oxide) of the following formula:

[0172]

[0173] -MAD463 (6-fluoro-2-phenylbenzo[d][1,2]selenazol-3(2H)-one 1-oxide) of the following formula:

[0174]

[0175] Preferred compounds according to the invention are ebselen and ebselen oxide, advantageously ebselen, and their derivatives or analogs, as defined below.

[0176] Ebselen (2-phenyl-1,2-benzoselenazol-3(2H)-one; CAS No. 60940-34-3), also known as PZ 51, DR3305 and SPI-1005, is a synthetic molecule that mimics glutathione peroxidase. Therapeutic activity of this molecule has been described with regard to ischemic lesions, stroke, hearing loss and bipolar disorder. It is also effective in treating Clostridioides difficile infections and provides fungicidal activity against Aspergillus fumigatus.

[0177] In practice, this molecule is already the subject of a Phase III clinical trial for the treatment of Meniere's disease (hearing impairment).Ebselen is administered orally in varying doses ranging from 200 mg to 600 mg (in the form of 200 mg gel capsules) per day for at least 21 days.

[0178] The present invention is also directed to derivatives or analogs of these compounds, in particular derivatives or analogs of ebselen, which have the same biological activities, i.e. those reported in the examples, in particular restoration of respiratory growth, restoration of cellular respiration or levels of mitochondrial ATP, whether in a yeast model (in particular Saccharomyces cerevisiae) of a filamentous mold (in particular the filamentous fungus Podospora anserina), or on human cells (in particular fibroblasts or myoblasts from a patient suffering from a disease condition).

[0179] The term "derivative" (or "analog") encompasses derivatives and metabolites and pharmaceutically acceptable salts. A derivative is a compound obtained from another compound (a precursor having a typically similar chemical structure) after transformation thereof. Derivatives may differ by one or more atoms or functional groups. Metabolites are stable intermediate compounds or compounds produced by biochemical transformation of an initial molecule through metabolism.

[0180] "Pharmaceutically acceptable salts" refer to addition salts of compounds that can be obtained by reaction of the compound with an inorganic or organic acid according to methods known per se. Among the acids traditionally used for this, mention may be made of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, 4-toluenesulfonic acid, methanesulfonic acid, cyclohexylaminosulfonic acid, oxalic acid, succinic acid, formic acid, fumaric acid, maleic acid, citric acid, aspartic acid, cinnamic acid, lactic acid, glutamic acid and N-acetylaspartic acid, N-acetylglutamic acid, ascorbic acid, malic acid, benzoic acid, nicotinic acid and acetic acid. According to a preferred embodiment, the compound is not in the form of a salt.

[0181] The compounds, in particular ebselen, may be modified to increase their stability, their bioavailability and / or their ability to reach target tissues, in particular mitochondria.

[0182] In a manner known to those skilled in the art, the compound, in particular ebselen, can be present in the composition in naked form (free), or contained in a delivery system that increases stability, targeting and / or bioavailability, such as liposomes, or incorporated into a carrier, such as a hydrogel, cyclodextrin, biodegradable nanocapsule, bioadhesive microspheres, carrier or in combination with a cationic peptide.

[0183] The invention also relates to pharmaceutical compositions comprising as active ingredient at least one compound as defined above, and to the use of said compounds or said compositions as a medicament or a pharmaceutical product.

[0184] Therefore, the present invention is directed to a pharmaceutical composition comprising a compound according to the present invention for targeted use. Advantageously, the composition comprises a therapeutically effective amount of the compound and a pharmaceutically acceptable carrier. In a specific embodiment, the term "pharmaceutically acceptable" refers to being approved by a regulatory agency from a federal or state government or registered in the U.S. or European Pharmacopoeia generally considered to be used for animals and humans. The term "carrier" refers to a diluent, adjuvant, excipient or vehicle administered together with a therapeutic product. The drug carrier can be a sterile liquid such as water and oil, including water and oil from petroleum sources, animals, plants or synthesis, such as peanut oil, soybean oil, sesame oil, etc. Saline solutions and aqueous solutions of glucose and glycerol can also be used as liquid carriers, particularly for injection solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, water, ethanol, etc.

[0185] If necessary, the composition may also contain a small amount of a wetting agent or emulsifier or pH buffer. The composition may take the form of a solution, suspension, emulsion, extended release formulation, etc. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin. The composition comprises a therapeutically effective amount of a therapeutic agent, preferably in a purified form, and an appropriate amount of a carrier to provide a form for adequate administration to a subject.

[0186] In a preferred embodiment, the composition is formulated according to conventional procedures (e.g., a suitable pharmaceutical composition, e.g., administered orally to humans). Typically, the composition for oral administration is in the form of a tablet, possibly a divisible tablet, or an effervescent tablet, and contains excipients suitable for solid dosage forms and for administration to humans. For example, ebselen can be in the form of a powder packaged in a tablet that can contain 200 mg of the active ingredient. The tablet can be crushed and mixed with a liquid.

[0187] Alternatively, the composition may be in liquid form, advantageously an aqueous composition. Any other suitable solvent may be used.

[0188] The amount of the therapeutic agent of the present invention (i.e., compound as described above) that is effective in the treatment of a disease can be determined by standard clinical techniques. In addition, in vivo and / or in vitro tests (such as those described in the following examples) may be used to help predict the range of the optimal dose. The exact dose used in the formulation will also depend on the route of administration, body weight, and severity of the disease, and must be determined according to the judgment of the practitioner and the condition of each patient.

[0189] According to a particular embodiment, the composition of the invention is in solid form, advantageously a tablet, which may contain 200 mg of the active compound, in particular ebselen.

[0190] According to one embodiment, the composition of the invention is in liquid form and advantageously comprises 30 μM, 20 μM or even 15 μM of the active compound, in particular ebselen. According to one embodiment, the composition of the invention is in liquid form and advantageously comprises 1 nM or more, even 10 nM or more, 30 nM or more, even 100 nM or more of the active compound, in particular ebselen, wherein for example the optimal concentration of the active compound, in particular ebselen, is 1 μM.

[0191] The appropriate route of administration must be such that a therapeutically effective amount of the therapeutic product is delivered to the target tissue, depending on the disease.

[0192] Ebselen and its derivatives can be administered in a pharmaceutically acceptable form by one of the administration routes known for active ingredients of this type.

[0193] The available routes of administration are as follows: topical (regional), enteral (systemic effect, but delivered via the gastrointestinal (GI) tract), or parenteral (systemic effect, but delivered via 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 administration, preferably oral administration.

[0194] According to other embodiments, administration can be parenteral, in particular by the intramuscular route (i.e. in the muscle) or systemic (i.e. in the circulatory system). In this context, the term "injection" (or "infusion" or "infusion") encompasses intravascular administration, in particular intravenous (IV) and intramuscular (IM) administration. Injection is usually performed using a syringe or catheter.

[0195] According to one embodiment, the composition is administered via the oral, intramuscular, intraperitoneal, subcutaneous, topical, regional or intravascular route, advantageously via the oral route.According to a preferred embodiment, the composition is intended for oral administration.

[0196] The 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 or tablets.

[0197] Alternatively, the container may be in the form of a liquid medicament such as an elixir or suspension containing various masking substances for color, flavor and stability.

[0198] According to a preferred embodiment, the composition is intended for oral administration. Advantageously, the composition is administered orally, ie via the mouth.

[0199] Preferably, the composition according to the invention is administered orally, in particular in the form of a gel capsule, a capsule or a tablet.

[0200] In order to produce oral galenic dosage forms according to the invention, in particular gel capsules, 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. If necessary, magnesium stearate as an additive provides a useful lubricating function.

[0201] In certain circumstances it may be advantageous to provide a controlled release form, particularly extended release via the known galenic dosage form.

[0202] Furthermore, the composition according to the invention may be intended for the preparation of a pharmaceutical composition which can be administered by injection.

[0203] The pharmaceutical composition according to the present invention can be dissolved or suspended in a pharmaceutically acceptable sterile injection liquid (such as sterile water, a sterile organic solvent or a mixture of the two liquids) for intravenous administration.

[0204] 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 drug delivery systems may also be used.

[0205] As already mentioned, the 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 or tablets.

[0206] The gel capsule or tablet can be taken with a small amount of water before or during the main meal. According to a preferred embodiment, the composition according to the present invention is administered daily, for example once a day, or even twice a day or more. Treatment can last for many weeks, months, years or even lifelong.

[0207] Typically, the dosage of the therapeutic agent (i.e., ebselen or one of its derivatives) varies according to factors such as the subject's age, weight, height, sex, general health, and medical preexisting conditions. Typically, it is desirable to provide a patient with a single dose of a therapeutic agent that is effective and non-toxic.

[0208] According to a specific embodiment of the invention, the dosage of the composition (advantageously a daily dose for oral administration to humans) is less than or equal to 10 mg 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.

[0209] As already indicated, the patient is advantageously a human being, in particular a neonate, infant, child, adolescent or adult, regardless of sex. However, the treatment tool according to the invention can be adapted and used to treat other animals, in particular pigs, mice, dogs or macaques.

[0210] In general, the present invention generally relates to the treatment of mitochondrial diseases, ie diseases associated with or caused by mitochondrial dysfunction. In the framework of the present application, the expression "diseases associated with mitochondrial dysfunction" is used to cover all such situations.

[0211] With regard to the following examples demonstrating the positive influence of ebselen or one of its derivatives on the mitochondrial respiratory chain, the diseases that present a particular interest are diseases of the mitochondrial respiratory chain.

[0212] Several mitochondrial diseases have been documented in the prior art:

[0213] NARP syndrome (neuropathy, ataxia and retinitis pigmentosa) induces a variable combination of developmental delay, retinitis pigmentosa, dementia, seizures, ataxia, proximal neurogenic myasthenia and sensory neuropathy. The NARP syndrome is caused by multiple mutations in the mitochondrial gene ATP6, which encodes the subunit α of the ATPase (complex V of the OXPHOS system). Mutations usually show heteroplasmy (coexistence of mutant and wild-type (WT) mitochondrial DNA (mtDNA) in the same cell). Depending on the type of mutation and the percentage of mutated mtDNA (the degree of heteroplasmy), clinical outcomes have variable severity. ATP6: m.8993T>C / G mutations are the most common in NARP patients with severe forms of NARP syndrome. FMC1 is a nuclear gene that encodes a protein required for assembling segment F1 of ATP synthase at high temperatures (35-37°C), thereby simulating the heteroplasmy observed in patients with NARP. Indeed, when cultivated at restrictive temperature (35-37°C), the mitochondria of the mutant fmc1Δ contained much fewer assembled ATP synthase complexes than the wild-type (WT) strain, but those that were assembled were fully functional. This heteroplasmy is also found in patients with reduced ATP synthase levels due to heteroplasmic mutations in the ATP6 gene. Therefore, the mutant fmc1Δ constitutes an appropriate model for this disease, in particular equivalent to the mutant m.8993T <G(MR14,NARP)(Schon EAet al.(2001)Cell DevBiol.12(6):441-8)。

[0214] The TAZ gene encodes tafazzin, a mitochondrial transcylase that catalyzes the remodeling of immature cardiolipin to its mature composition, which mainly contains tetralinoleyl fragments. <3980> TAZ<!--3980--> Mutations cause Barth syndrome, an X-linked disorder typically characterized by dilated cardiomyopathy (CMD) with endocardial fibroelastosis (EFE), proximal-predominant skeletal myopathy, growth failure, neutropenia, and organic aciduria, particularly excess 3-methylglutaconate (Barth, PGet al. (1996) J Inherit Metab Dis, 19, 157-160).

[0215] The genes COX2 and SURF1 encode a subunit and an assembly factor of mitochondrial complex IV, respectively. Mutations are associated with Leigh syndrome, a severe progressive neurodegenerative disease that appears in the first month and year of life and causes early death. Affected individuals typically have global developmental delay or regression, hypotonia, ataxia, dystonia, and ophthalmological abnormalities, such as nystagmus or optic atrophy (Barrientos, A. et al. (2002) EMBO J, 21, 43-52).

[0216] MPV17 encodes a protein of the inner mitochondrial membrane whose function is unknown. Mutations in MPV17 cause:

[0217] - Mitochondrial-6 DNA depletion syndrome, a recessive autosomal disorder characterized by progressive hepatic insufficiency in infants, often leading to death in the first year of life. Those infants who survive develop progressive neurological impairment, particularly ataxia, hypotonia, dystonia, and psychomotor regression (Spinazzola, A. et al. (2006) Nat Genet, 38, 570-575).

[0218] -Navajo type neuropathy: manifestations include corneal ulcers caused by severe anesthesia, non-painful fractures, extremity mutilations, muscle weakness, and absent or significantly reduced deep tendon reflexes in the absence of intellectual disability (Karadimas, CL et al. (2006) Am J Hum Genet, 79, 544-548).

[0219] Of particular interest is the treatment of diseases selected from the group consisting of MELAS syndrome, myopathy or cardiomyopathy inherited from the mother, NARP or MILS syndrome, Leigh syndrome, Leber hereditary optic neuropathy (LHON), Barthel syndrome, mitochondrial DNA depletion syndromes, particularly 4A (Alpers type) and 4B (MNGIE type), mitochondrial recessive ataxia syndromes, sensory ataxic neuropathy, dysarthria and ophthalmoplegia, spinocerebellar ataxia with epilepsy, progressive external ophthalmoplegia, mitochondrial DNA-6 depletion syndrome, Navajo type neuropathy, Behr syndrome, mitochondrial DNA 14 depletion syndrome, infantile cardioencephalomyopathy due to lack of cytochrome C oxidase (COA6 mutation), mitochondrial type 1 complex III nuclear defect, GRACILE syndrome and Bjornstad syndrome.

[0220] Of particular interest are the treatments of diseases associated with defects in one or more mitochondrial complex I. Certain diseases are uniquely associated with dysfunction of complex I, while other diseases are associated with multiple defects, for example affecting multiple mitochondrial complexes.

[0221] In a known manner, the respiratory chain in mitochondria involves oxidative phosphorylation, an important cellular process that uses oxygen and monosaccharides to produce adenosine triphosphate (ATP), the main energy source of the cell. Five protein complexes that constitute the OXPHOS system are involved in this process, each of which is composed of a variety of proteins. The complexes are respectively called Complex I, Complex II, Complex III, Complex IV and Complex V. Complex I (CI or NADH dehydrogenase or NADH coenzyme Q reductase), the first enzyme of the respiratory chain, is a very large protein complex (about 1000 kDa) consisting of at least 44 subunits including 7 subunits encoded by mitochondrial DNA (ND1 to ND6 and ND4L).

[0222] According to a specific embodiment, the compounds according to the invention, in particular ebselen, can be used to treat so-called "primary" mitochondrial diseases, i.e. diseases due to identified genetic abnormalities associated with one or more mitochondrial or nuclear DNA mutations identified in at least one subunit of the OXPHOS complex, in particular Complex I. These conditions are associated with neurological, myocardial or ophthalmological symptoms, the symptoms being related to the tissue or organ most affected in these mitochondrial diseases, even though other organs or tissues may be affected.

[0223] According to a specific embodiment, the compounds according to the invention, in particular ebselen, can be used to treat so-called "secondary" mitochondrial diseases. In this case, the genetic abnormality does not directly involve the OXPHOS complex, but the condition will affect mitochondrial function and in particular can lead to a reduction in the activity of the enzymes of this process. Such diseases can also be due to 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.

[0224] According to a preferred embodiment, the defect or mitochondrial dysfunction is caused by a genetic disease.

[0225] By definition, a genetic disease is a disease caused by one or more genetic defects (abnormalities) (or mutations) in one or more genes. The genetic defects can affect mitochondrial DNA and / or nuclear genes. The genetic defects that cause mitochondrial diseases can be isolated mutations that lead to codon changes. However, the disease may be associated with a deletion or insertion of one or more bases or codons.

[0226] According to a specific embodiment, the disease is caused by one or more genetic defects (or mutations) in one or more genes involved in the function of the OXPHOS complex, in particular Complex I.

[0227] A non-limiting list of such genes includes:

[0228] - 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, NDUFB1 1. NDUFC1, NDUFC2, NDUFS1, NDUFS2, NDUFS3, NDUFS4, NDUFS5, NDUFS6, NDUFS7, NDUFS8, NDUFV1, NDUFV2, NDUFV3;

[0229] - Complex I assembly genes, in particular NDUFAF1, NDUFAF2, NDUFAF3, NDUFAF4, NDUFAF5, NDUFAF6, NDUFAF7, NDUFAF8, NUBPL, ACAD9, TMEM70, TMEM126B, FOXRED1, ECSIT, AIF, TIMMDC1.

[0230] According to a specific embodiment, the disease to be treated in the framework of the present invention is associated with or is caused by at least one genetic defect or mutation in at least one of the following genes: MTTL1, ATP6, FMC1, TAZ, COX2, SURF1, POLG, MPV17, OPA1, COA6, ND6 and BCS1L, advantageously ATP6, TAZ, COX2, SURF1.

[0231] Several mitochondrial diseases of genetic origin have been documented in the prior art, particularly those associated with defects in mitochondrial complex I:

[0232] LHON syndrome or Leber hereditary optic neuropathy usually presents in young people. The onset is sudden, with a rapid decrease in vision in the center of the eye, which corresponds to a decrease in central visual acuity. Most often, peripheral vision is preserved as a visual halo around the blind area. The disease is due to common homoplasmic mutations in the genes encoding subunits of complex I of the respiratory chain. In practice, the mitochondrial DNA mutations m.11778G>A, m.3460G>A and m.14484T>C account for about 95% of LHON mutations.

[0233] Leigh syndrome (or LS) is a serious neurodegenerative disease. Affected people usually have overall developmental delay or developmental regression, hypotonia, ataxia, dystonia and ophthalmological abnormalities, such as nystagmus or optic atrophy. Leigh syndrome may also have harmful multi-system effects on the heart, liver, gastrointestinal and kidney organs. Biochemical studies of patients with Leigh syndrome have a trend of increased lactate and abnormal mitochondrial oxidative phosphorylation. Leigh syndrome can be associated with mutations in genes encoding subunits of complex I, such as mutation NDUFV1 or mutation MTND5 m.13513G>A.

[0234] MELAS syndrome, which includes mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-type episodes, is a genetically heterogeneous mitochondrial disorder with a variable clinical phenotype. The disorder is accompanied by features of central nervous system damage, particularly seizures, hemiparesis, hemianopia, cortical blindness, and occasional vomiting. The syndrome was first associated with mitochondrial DNA, namely the gene tRNA Leu(UUR) The MELAS syndrome is associated with a mutation in mitochondrial DNA (MTTL1), m.3243A>G, which induces changes in the translation of mRNAs for complex I and thus induces a reduction in the amount of structural proteins of complex I, such as the mitochondrial subunit ND6. MELAS syndrome can also be associated with other mitochondrial DNA mutations, such as the mutation m.3260A>G, which also affects tRNA Leu(UUR) The mutation m.3260A>G may also lead to other clinical phenotypes, especially myopathy and cardiomyopathy of maternal origin.

[0235] Therefore, the treatment of genetic diseases that have been shown to be associated with complex I defects, such as MELAS syndrome, Leigh's syndrome and Leber's hereditary optic neuropathy (LHON), is of particular interest. It should be noted that such diseases may be associated with other symptoms such as cardiac, myopathic or neurological clinical phenotypes.

[0236] More generally, the compounds according to the invention, in particular ebselen, can be used to treat mitochondrial dysfunction, in particular mitochondrial dysfunction associated with defects in Complex I. Mitochondrial dysfunction, characterized by a loss of efficiency of the electron transport chain and a reduced synthesis of high-energy molecules such as adenosine-5'-triphosphate (ATP), is a characteristic of aging and generally of all chronic diseases.

[0237] Such diseases include:

[0238] - Neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (Lou Gehrig's disease), and Friedreich's ataxia;

[0239] - Cardiovascular diseases, such as atherosclerosis and other heart and blood vessel diseases;

[0240] - Diabetes and metabolic syndrome;

[0241] -Autoimmune diseases such as multiple sclerosis, systemic lupus erythematosus, and type 1 diabetes;

[0242] - Neurobehavioral and psychiatric disorders, such as autism spectrum disorder, schizophrenia, bipolar and mood disorders;

[0243] - Gastrointestinal disorders;

[0244] - Fatigue disorders such as chronic fatigue syndrome and Gulf War illness;

[0245] -Musculoskeletal disorders such as fibromyalgia and hypertrophy / atrophy of skeletal muscles;

[0246] - Muscular dystrophy,

[0247] - cancer; and

[0248] - Chronic infection.

[0249] The treatment proposed by the present invention (based on ebselen or one of its derivatives) can also be combined with other treatments aimed at treating the same condition or the same disorder or another disease.

[0250] According to a particular embodiment, said additional treatment is based on the administration of another compound of interest.

[0251] 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 properties and in particular their bioavailability.

[0252] According to a specific application mode, the present invention relates to a composition, advantageously a pharmaceutical composition or a medicament, comprising a compound as described above and other active molecules (other genetic therapy molecules, chemical moieties, peptides or proteins) potentially useful for treating the same disease or a different disease (advantageously the same disease).

[0253] Preferably, the pharmaceutical composition according to the present invention and at least one compound for treating the same disease or different diseases are administered simultaneously, separately or distributed over time for treating the same disease or different diseases.

[0254] More generally, with respect to mitochondrial diseases, another compound capable of improving mitochondrial function may be administered simultaneously or at different times. In the case of simultaneous administration, the two compounds may be combined in the same composition.

[0255] Examples of such another compound 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 (vitamin B2) reduced in nicotinamide adenine dinucleotide (NADH), L-arginine, possibly in combination.

[0256] Examples of compounds used, for example in the case of MELAS syndrome, are nitric oxide (NO) precursors such as L-arginine and citrulline.

[0257] According to a specific embodiment and in view of the examples demonstrating a potential synergistic effect, the 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 diethyldithiocarbamate, alverine and alverine citrate.

[0258] Subjects who may benefit from the composition according to the present invention include all patients suffering from a disease associated with mitochondrial dysfunction, in particular a disease associated with Complex I deficiency, who are diagnosed as carriers of such a disease or are at risk of developing such a disease.

[0259] The subject to be treated with the composition according to the invention can be selected based on different criteria. Regarding mitochondrial dysfunction, in particular Complex I deficiency, various tests can be performed, such as:

[0260] - At the biochemical level, O2 consumption and / or mitochondrial complex I activity can be measured from a biopsy from the subject, particularly muscle or skin. The activity of other complexes of the respiratory chain can also be assessed to determine if mitochondrial dysfunction is due solely to a complex I defect:

[0261] - At the genetic level: Sequencing of mitochondrial or nuclear DNA extracted from blood, from cells or from a biopsy sample (e.g. from the skin) will allow the identification of one or more molecular abnormalities, in particular mutations or deletions / insertions in genes, in particular those listed below. Alternatively, the expression or activity of the corresponding protein may be assessed by any method known to those skilled in the art (e.g. by Western blotting).

[0262] It is an object of the present invention to provide a safe (non-toxic) treatment.Another object is to provide an effective treatment that will allow delaying, slowing or preventing the progression of the disease and possibly improving the patient's phenotype, which can be monitored at the clinical level as shown below.

[0263] For a subject, the composition according to the present invention can be used for:

[0264] - Improve mitochondrial function, especially mitochondrial respiration;

[0265] - Improved growth;

[0266] - Improve muscle function;

[0267] - Improved vision and / or hearing;

[0268] - Improve respiratory function;

[0269] - Improve heart, liver or kidney function;

[0270] - Improve brain function;

[0271] - Improve digestive function; and / or

[0272] -Prolonging survival, more generally improving quality of life and life expectancy.

[0273] According to one aspect, the present invention relates to a method for improving mitochondrial function, in particular Complex I activity, advantageously without undesired effects, comprising administering to a subject in need thereof a therapeutic amount of a composition as described above.

[0274] Advantageously, the improvements are observed up to 1 month, or 3 months, or 6 months, or 9 months after initiation of treatment, more advantageously up to 1 year, 2 years, 5 years, 10 years, or even throughout the subject's lifetime after initiation of treatment.

[0275] In one embodiment, the improvement is manifested as a decrease in severity and / or frequency and / or a delay in the onset of symptoms.

[0276] Improvement can be assessed based on methods known in the art, for example in the case of MELAS:

[0277] - assessment of the amount of lactate, in particular cerebral ventricular lactate, for example measured by magnetic resonance spectroscopy (MRS);

[0278] - Assessment of quality of life and / or expectations by clinical rating scales, such as the NMDAS (Newcastle Mitochondrial Disease Scale for Adults) score or the SF-36 (Short Form Health Survey) score;

[0279] - Assessing brain changes, for example using magnetic resonance imaging (MRI);

[0280] - Assessment of changes in muscle activity through physical testing (e.g., six-minute walk test);

[0281] -Assess changes in venous lactate and GDF15 concentrations;

[0282] -Evaluation of mtDNA heteroplasmic modifications in urine and blood.

[0283] Appropriate parameters for a given case can be adjusted depending on the disease.

[0284] Therefore, the claimed treatment allows to improve the clinical conditions and different parameters disclosed above relative to untreated subjects.

[0285] Unless otherwise indicated, embodiments of the present invention employ conventional techniques of microbiology, cell biology, biochemistry, and immunology and molecular biology (including recombinant techniques), which are known to those skilled in the art. These techniques are explained in detail in the literature, especially 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). Particularly useful techniques for specific embodiments will be discussed in the following sections.

[0286] The disclosures of the patents, patent applications, and publications cited in this application are hereby incorporated by reference in their entirety.

[0287] Without further description, it is believed that one skilled in the art can make and use the compounds of the invention and practice the claimed methods by using the description and following exemplary embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0288] The invention and the advantages resulting from it will become clearer through the following examples of implementation supported by the related drawings. These, however, do not have any limiting effect.

[0289] Figure 1:

[0290] A / Effects of ebselen (EBS) on respiratory growth in various yeast models of mitochondrial disease on non-fermentable solid media as measured by the halo test

[0291] C / Effect of ebselen oxide on respiratory growth of mutant yeast strain taz1 on non-fermentable solid medium (detected by halo test) compared with ebselen (B) and DMSO (D).

[0292] Effects of E / ebselen (EBS) and different analogs of ethaselen on the respiratory growth of yeast strains taz1, fmc1, cox2, and A29G.

[0293] Figure 2 :Effects of ebselen (EBS) on mutant yeast strain taz1

[0294] A / Respiratory growth in liquid culture

[0295] Accumulation of B / Cox2 protein

[0296] Figure 3 :Effects of ebselen (EBS) on mutant yeast strain sym1

[0297] A / Respiratory growth in liquid culture

[0298] B / Cellular respiration

[0299] Accumulation of C / Cox2 protein

[0300] Figure 4 : Model for EBS in cells carrying mutations in mitochondrial complex I (Pa nuo-51 A357V )'s thermosensitive growth

[0301] A / Effects on Growth Halo

[0302] B / Dose-dependent effects on growth

[0303] Figure 5: Effects of EBS on proliferation and viability of TAZ patient cells

[0304] A / Growth of fibroblasts from TAZ patients in the presence or absence of EBS (12.15 nM)

[0305] B / Changes in respiratory growth of fibroblasts from TAZ patients at 96 hours as a function of EBS concentration (from 150 pM to 234.9 μM) compared to the negative control DMSO.

[0306] C / Changes in respiratory growth of fibroblasts from TAZ patients at 96 hours as a function of ethaselenin concentrations (from 150 pM to 26.1 μM) compared to the negative control DMSO.

[0307] D / Levels of mitochondrial ATP in fibroblasts from TAZ patients incubated in the presence of antimycin A (100 μM), rotenone (0.5 μM) or oligomycin A (3 μM) and EBS at a concentration of 12.15 nM.

[0308] Respiratory growth of E / TAZ patient myoblasts at 120 hours as a function of EBS concentration (from 50 pM to 234.9 μM).

[0309] FIG. 6 : Effect of EBS on the proliferation and viability of HeLa cell lines in which the FMC1 gene was deleted in 1 g / L glucose (A) or galactose (B) medium

[0310] DO(AU): Optical density and arbitrary units

[0311] Figure 7 :Effect of EBS on mitochondrial respiration in cells from patients with complex I (CI) deficiency of the mitochondrial respiratory chain (carrying mutations in the nuclear gene NDUFV1 that affects the complex)

[0312] A / Oxygen Consumption Rate (OCR)

[0313] B / Basic breathing (normal)

[0314] C / Respiration associated with ATP synthase (RO)

[0315] D / maximum breathing

[0316] Figure 8 :Synergistic effects among ebselen (EBS), disulfiram (DSF) and alverine (ALV)

[0317] A / In the taz yeast mutant model

[0318] B / In the mutant Pa nuo-51 A357V Middle: Growth rate in the presence of EBS and ALV alone or in combination compared to the negative control DMSO (VEH)

[0319] Fig. 9 : Effect of EBS administration on the weight of KO mice lacking the TAZ gene (a mouse model for Barthel syndrome). DETAILED DESCRIPTION

[0320] Examples 1 to 3: Saccharomyces cerevisiae model

[0321] The strains of Saccharomyces cerevisiae used in these examples each contain different specific mutations that model human mutations that cause mitochondrial disease. To varying degrees, all of these mutant yeast strains have growth defects when cultured on respiratory media such as ethanol or glycerol at 28°C or 36°C (depending on the strain).

[0322] Mutated yeast strains:

[0323] -fmc1:MC6 MATa ade2–1his3–11,15trp1–1leu2–3,112ura3–1fmc1::HIS3

[0324] [Δi ER OR] (Schwimmer, C. et al. (2005) J Biol Chem, 280, 30751-30759) was used for the first screening

[0325] -MR14 MAT a ade2-1 his3-11,15trp1-1 leu2-3,112ura3-1 CAN1 arg8::HIS3ρ+atp6-L173R or RKY20 MATa ade2-1 his3-11,15trp1-1 leu2-3,112ura3-1CAN1 arg8::HIS3ρ+atp6-L173P (Rak, M. et al. (2007) J Biol Chem, 282, 34039-34047) was used for the second screening

[0326] -taz1Δ: This strain was prepared by replacing the open reading frame of TAZ1 with the open reading frame of TRP1 in strain W303-1A (MATa ade2-1 ura3-1 his311, 15trp1-1 leu2-3, 112can1-100) (from Taffin de Tilques, M. et al. (2017) Dis Model Mech, 10, 439-450)

[0327] -shy1-G137R: This mutant was produced in strain CW252, which contains the W303 nucleobase and a mitochondrial gene lacking introns. This gene is a homolog of the human gene SURF1

[0328] -sym1Δ: This yeast strain was produced by replacing the open reading frame of SYM1 with the open reading frame of kanMX6 in strain W303-1A (MATa ade2-1 ura3-1 his311,15trp1-1leu2-3,112can1-100). This gene is a homolog of the human gene MPV17

[0329] -cox2: MATa lys2 leu-2-3,112ura3-52 his3ΔHindIII arg8::hisG cox2-22 (Bonnefoy, N. et al. (2001) Mol Cell Biol, 21, 2359-2372.

[0330] -A29G: The yeast strain used in these experiments included tRNA-Leu A29G with the genetic bases MATα, his3–11, ade2–1, leu2–3, 112, ura3–1, trp1-D2, can1–100, syn.

[0331] Related mitochondrial diseases:

[0332] NARP syndrome: NARP syndrome (neuropathy, ataxia, and retinitis pigmentosa) causes a variable combination of developmental delay, retinitis pigmentosa, dementia, seizures, ataxia, proximal neurogenic weakness, and sensory neuropathy. NARP is caused by various mutations in the mitochondrial gene ATP6, which encodes the subunit α of the ATPase (complex V of the OXPHOS system). Mutations often show heteroplasmy (coexistence of mutant and wild-type (WT) mitochondrial DNA (mtDNA) in the same cell). Depending on the type of mutation and the percentage of mutated mtDNA (the degree of heteroplasmy), the clinical outcome is of variable severity. The ATP6:m.8993T>C / G mutation is the most common in patients with NARP, which causes the severe form of NARP syndrome. FMC1 is a nuclear gene that encodes a protein required for the assembly of segment F1 of the ATP synthase at high temperatures (35-37°C), thereby mimicking the heteroplasmy observed in patients with NARP. Indeed, when cultured at restrictive temperature (35-37°C), the mitochondria of the mutant fmc1Δ contained much fewer assembled ATP synthase complexes than the wild-type (WT) strain, but those that were assembled were fully functional. This heterogeneity has also been found in patients with reduced ATP synthase levels due to heteroplasmic mutations in the ATP6 gene. Therefore, the mutant fmc1Δ constitutes a suitable model for the disorder (Lefebvre-Legendre, L. et al. (2001) J Biol Chem, 276, 6789-6796). Various NARP yeast strains carrying homoplasmic mutations equivalent to mutations T8993G and T8993C (Rak, M. et al., J Biol Chem. 2007 282 (47): 34039-47) were also tested.

[0333] TAZ

[0334] The TAZ gene encodes tafazzin, a mitochondrial translocase that catalyzes the remodeling of immature cardiolipin to its mature composition, which mainly contains tetralinoleyl fragments. <6920> TAZ<!--6920--> Mutations cause Barth syndrome, an X-linked disorder typically characterized by dilated cardiomyopathy (CMD) with endocardial fibroelastosis (EFE), proximal-predominant skeletal myopathy, growth failure, neutropenia, and organic aciduria, particularly excess 3-methylglutaconate (Barth, PGet al. (1996) J Inherit Metab Dis, 19, 157-160).

[0335] COX2 and SURF1

[0336] The genes COX2 and SURF1 (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 disease that appears in the first month and year of life and can lead to early death. Affected people often have global developmental delay or regression, hypotonia, ataxia, dystonia, and ophthalmological abnormalities, such as nystagmus or optic atrophy (Barrientos, A. et al. (2002) EMBO J, 21, 43-52).

[0337] MPV17: MPV17 (SYM1 gene in yeast) encodes a protein of the inner mitochondrial membrane whose function is unknown. Mutations in MPV17 cause:

[0338] - Mitochondrial-6 DNA depletion syndrome, a recessive autosomal disorder characterized by progressive hepatic insufficiency in infants, often leading to death in the first year of life. Those infants who survive develop progressive neurological impairment, particularly ataxia, hypotonia, dystonia, and psychomotor regression (Spinazzola, A. et al. (2006) Nat Genet, 38, 570-575).

[0339] - Navajo type neuropathy: manifestations include severe sensorimotor peripheral neuropathy with liver and brain damage, such as leukoencephalopathy and corneal ulcers. (Karadimas, CL et al. (2006) Am J Hum Genet, 79, 544-548).

[0340] - MELAS syndrome: The mutation A30(29)G in yeast A29G mimics the human mutation m.3260A>G in the gene tRNALeu(UUR) that causes MELAS syndrome.

[0341] Example 1: Effects of Ebselen (EBS) and its derivatives on mutant yeast strains cultured in a non-fermentable (respiratory) environment

[0342] Materials and methods

[0343] As previously described (WO2020 / 254632), different mutant yeast strains were spread on solid respiring agar-agar based media (containing glycerol or ethanol as the sole carbon source) and exposed to filters on which the tested compounds (particularly ebselen (EBS)) were deposited. These plates were then incubated at the indicated temperature (either 28°C or 36°C, depending on the strain used).

[0344] More specifically, exponentially growing mutant cells at 0.125 OD were evenly spread with sterile glass beads in square culture dishes (12 cm x 12 cm) containing solid respiration medium (YPA ethanol, 1% yeast extract, 0.5% Bacto Peptone, 40 mg / L adenine, 2% ethanol for sym1 and taz1; or 2% YPA glycerol for cox2, fmc1, atp6, shy1 et A29G). Small sterile filters were then placed on an agar-agar surface and the compounds tested at the growth concentrations were added to the filters in the amounts shown (30 and 90 nmol). On each culture dish, DMSO (buffer) was used as a negative control (upper left filter). The culture dishes were then incubated at 28°C (for shy1) or 36°C (for cox2, sym1, fmc1, atp6, taz1 and A29G) for 4 to 5 days and then photographed. After multiple days of incubation, the improvement in growth was assessed by the appearance of a halo of improved growth around the filter where the active ingredient was deposited.

[0345] result

[0346] like Figure 1A As can be seen, EBS is active in a dose-dependent manner against all yeast mutant strains tested.

[0347] For mutant taz1, Figure 1C shows that ebselen oxide (100 nmol) has comparable activity to ebselen (100 nmol) (Figure 1B), while no halo was observed in the presence of DMSO (Figure 1D).

[0348] More generally, different analogs of EBS and ethaselen were evaluated for different mutants (taz1, fmc1, cox2, and A29G). Figure 1E As shown, all analogs tested were active against the strains tested.

[0349] Example 2: Effect of Ebselen (EBS) on mutant yeast strain taz1

[0350] Materials and methods

[0351] Respiratory growth in liquid culture:

[0352] To determine the optimal concentration of EBS that results in suppression of the respiratory growth defect of mutant yeast strains, cells undergoing exponential growth were inoculated in fresh ethanol YPA non-fermentable medium supplemented with or without increasing concentrations of EBS. Cell growth in liquid respiratory medium was monitored over a period of 80 h using a bioscreen system, during which (OD600nm) cell density was sampled every 20 minutes.

[0353] Cellular Respiration:

[0354] Respiration intensity corresponds to the amount of oxygen consumed relative to time and cell number. It reflects the mitochondrial oxidative metabolism of the cells. Oxygen consumption was measured using the OROBOROS oxygen graph system. Cells were cultured for 7-8 generations at 28°C in ethanolic YPA medium supplemented with DMSO or EBS (1 μM) for 24 to 48 h. 7 Oxygen profiles were introduced into each cell. O2 consumption was recorded with or without CCCP (maximum rate of oxygen consumption). The rate of O2 consumption was calculated based on the linear portion of O2 consumption.

[0355] Accumulation of Cox2 protein:

[0356] Total protein extracts from mutant yeast strains were analyzed by SDS-PAGE (50 μg / 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 were normalized to Ade13p and expressed relative to wild-type (WT) strains.

[0357] result

[0358] like Figure 2 As shown in A, on the mutant strain taz1, EBS was toxic starting at 27 μM and was active between 1 nM and 3 μM, with the optimal concentration being 1 μM.

[0359] In addition and if Figure 2 As shown in B, 1 μM EBS increased the accumulation of the protein Cox2, a subunit of complex IV of the mitochondrial respiratory chain, in the mutant strain taz1.

[0360] Example 3: Effect of Ebselen (EBS) on mutant yeast strain sym1

[0361] The experiments described in Example 2 were performed on the mutant yeast strain sym1.

[0362] The results are shown in Figure 3 middle.

[0363] Data are means ± SEM of at least three independent experiments. The significance of changes between samples and controls was assessed using multifactorial Anova: Tukey test.

[0364] For the mutant strain taz1, at the optimal concentration of EBS (1 μM), cellular respiration as well as the level of Cox2 increased compared to untreated cells.

[0365] Example 4: PODOSPORAANSERINA Model

[0366] The Saccharomyces cerevisiae used in Examples 1 to 3 does not possess complex I of the mitochondrial respiratory chain. Therefore, the EBS was tested on the strict filamentous mold Podospora.

[0367] The strain used in the following examples contains specific mutations that model human mutations in NDUFV1, a subunit of complex I that causes mitochondrial disease. This strain has a growth defect at permissive temperatures greater than 31.5°C.

[0368] Mutant strains of filamentous fungi Podospora:

[0369] nuo-51 A357V : The mutation A357V was introduced into the gene nuo-51 of the wild-type strain S together with the resistance cassette to nourseothricin (NatR). To simulate the human disease NDUFV1A341V, the genes NDI-1 and AOX were inactivated (El-Khoury et al., (2008) Curr Genet. 53: 249-58)

[0370] Genotype of strain: S, mat - ,nuo-51 A357V ,Δndi-1,Δaox,Nat R ,Hygro R

[0371] Materials and methods

[0372] The thermosensitive mutant Pa nuo-51 A357V Evenly spread on sterile glass beads on a square petri dish containing solid minimal medium (M2). Then a small sterile filter is placed on the agar-agar surface and 20nmol EBS is added. DMSO (the vehicle of the compound) is used as a negative control. The plate is then incubated at a non-permissive temperature of 33°C for 4-5 days and then photographed.

[0373] Then the mutant Pa nuo-51 was cultured at 32°C A357V , where their growth was reduced relative to the wild-type (WT) strain. The effect of EBS on the growth rate of the mutants was determined on supplemented minimal medium or in the absence of increasing EBS concentrations and estimated in centimeters of growth per day (cm / day).

[0374] result

[0375] Figure 4 A shows the positive effect of EBS on the growth of the mutant. More specifically, between 0.003 and 0.03 μM, EBS demonstrated that the mutant Pa nuo-51 A357V Significant improvement in growth ( Figure 4 B).

[0376] Example 5: Human cell model

[0377] Materials and methods

[0378] Statistical analyses were performed using the statistical test ANOVA with two mixed factors applying the Bonferroni multiple testing correction or using the GraphPad software Prism 9.5.1.

[0379] Stars indicate p values ​​< 0.05 (*), < 0.001 (**), < 0.0001 (***), < 0.0001 (****).

[0380] Fibroblasts and myoblasts from TAZ patients:

[0381] The effect of EBS on the respiratory growth of fibroblasts from TAZ patients was tested in comparison with fibroblasts from 8 different patients, carriers of TAZ gene mutations (with Barthel syndrome). Data show the percentage of growth compared to T0.

[0382] Proliferation rates were measured in 384-well microtiter plates (1200 cells / well at T0) in medium with a weak glucose concentration (1 g / L; to force cells to use the OXPHOS system instead of glycolysis) using a sulforhodamine B (SRB) staining protocol 24, 48, 72, or 96 hours after placing cells in the presence or absence of 12.15 nM EBS.

[0383] The proliferation rate was then measured under the same experimental conditions but using different concentrations of EBS: cells were analyzed 96 hours after treatment with increasing doses of EBS (from 150 pM to 234.9 μM).Data are the mean of 8 patients or 3 controls in triplicate in 4 independent experiments.

[0384] The proliferation rate was then measured under the same experimental conditions but with different concentrations of ethaselen: cells were analyzed 96 hours after treatment with increasing doses of ethaselen (from 150 pM to 26.1 μM). The bars represent the mean of 4 replicates from 2 patients in 3 independent experiments.

[0385] The Promega CellTiter-Glo kit was used to assess the level of mitochondrial ATP in fibroblasts from TAZ patients. Cells in 96-well plates (5000 / well) were treated or untreated with 12.15 nM EBS for 96 h and then incubated in the presence of antimycin A (100 μM), rotenone (0.5 μM), or oligomycin A (3 μM).

[0386] Also tested the ability of EBS to restore cell growth from myoblasts of patients (carriers of TAZ gene mutations). 48, 72, 96 or 120 hours after placing cells, using sulforhodamine B (SRB) staining scheme, in 384-well microtiter plates (at T0, 1200 cells / well) in a medium with a weak glucose concentration to measure proliferation rate. The figure shows the growth rate corresponding to the average value of 3 independent experiments at 120 hours.

[0387] KO ("knockout") cell line FMC1 :

[0388] The cell line HeLa KO FMC1, which mimics human diseases associated with ATP synthase deficiency (NARP, MILS syndrome), was generated by the CRISPR-Cas9 approach. The cell line HeLa KO FMC1 was cultured in DMEM medium containing 4.5 g / L glucose, 110 mg / l pyruvate and 50 mg / l uridine.

[0389] 96 h after seeding and treating cells, sulforhodamine B (SRB) staining protocol was used in 384-well microtiter plates (4500 cells / cm at T0). 2 The proliferation rate of this HeLa cell line in which the gene FMC1 was deleted was measured in a medium containing low concentrations of glucose (1 g / L) or galactose. The bars represent the average of 8 replicates from 3 independent experiments. The data show the growth percentage compared to T0.

[0390] KO ("knockout") cell line CI (Complex I) :

[0391] The ability of EBS to restore mitochondrial respiration in fibroblast cell lines from patients carrying mutations in the nuclear gene NDUFV1 of mitochondrial complex I was also tested. The human line NDUFV1, which mimics a human disease associated with mitochondrial defects in complex I (Leigh's syndrome), is a carrier of a genetic mutation affecting the gene NDUFV1. These cell lines were cultured in DMEM medium containing 1.0 g / L glucose and 1 mM sodium pyruvate and 50 μg / ml uridine (Sigma Aldrich, Lyon, France).

[0392] Respiration was measured using a Seahorse XF96 extracellular flow metabolic analyzer in 96-well plates according to the manufacturer's protocol (Agilent Technologies). Defective cells were exposed to different concentrations for 48 hours, with dose-response curves associated with a concentration range from 25 nM to 10 μM. (A) shows oxygen consumption rate, (B) basal respiration, (C) respiration associated with ATP synthase, and (D) maximum respiration. Bars represent the average of 4 independent replicates.

[0393] Primary fibroblasts in the exponential growth phase were detached by adding 2 ml of 0.05% trypsin. The fibroblasts were used to inoculate XF96 plates (30×10 3 The cells were then incubated in DMEM without bicarbonate (Sigma-Aldrich) supplemented with 5.5 mM glucose and 2 mM L-glutamine in a CO2-free incubator for 1 hour.

[0394] The measurement of oxygen consumption rate (OCR) is recorded under basal conditions to assess mitochondrial respiratory activity, and then successively treated cells with 4 μ g / mL oligomycin and 1.5 μM carbonyl cyanide to (trifluoromethoxy) phenylhydrazone (FCCP) (Sigma-Aldrich).Determine basal OCR, the respiration relevant to ATP, maximum respiration capacity and the respiration by proton leak.From all OCR measured values, subtract non-mitochondrial respiration (OCR after treatment with 2 μ g / ml antimycin A).From the difference between the basal respiration rate, assess the respiration (RO) relevant to ATP production, and inhibit by oligomycin, and obtain the respiration by proton leak by subtracting non-mitochondrial respiration from the ROC measured after oligomycin treatment.Maximum respiration capacity is determined as the respiration rate in the presence of uncoupling FCCP.Results are standardized according to cell number.

[0395] result

[0396] Figure 5A EBS at 12.15 nM was shown to improve respiratory growth of fibroblasts from TAZ patients. At this concentration, EBS was effective after 24 hours, and the improvement in cell proliferation was more pronounced at three other sampling times (48, 72 and 96 hours).

[0397] Figure 5B It was shown that EBS improved the respiratory growth of fibroblasts from TAZ patients over a wide concentration range at 96 hours: from 1.35 nM to 8.7 μM EBS had a beneficial effect relative to the negative control DMSO, with toxicity above 234.9 μM.

[0398] Figure 5C It was shown that ethaselen improved proliferation of cells from patients with Barthel syndrome over a wide range of concentrations from about 1 nM to 3 μM at 96 hours.

[0399] Figure 5D showed that EBS at a concentration of 12.15 nM improved mitochondrial ATP production in fibroblasts from TAZ patients.

[0400] exist Figure 5EThe effect of EBS on the restoration of growth of a myoblast cell line from a patient carrying a TAZ gene mutation is shown in : it shows that EBS from 450 pM to 8.7 μM has a beneficial effect compared to the negative control DMSO, where it is toxic above 78.3 μM.

[0401] Figure 6 shows that ebselen improves the proliferation of model cells of ATP synthase deficiency (NARP, MILS syndrome) in a dose-dependent manner and over a wide concentration range (pM to μM).

[0402] Figure 7 Ebselen was shown to improve respiration in model cells of a mitochondrial condition associated with Complex I from 25 nM to 100 nM and to be toxic above 10 μM.

[0403] Example 6: Synergistic effects between different compounds

[0404] Disulfiram (DSF) and alverine (ALV) are reported in documents WO2020 / 254632 and WO2022 / 018297, respectively, as being useful in treating conditions associated with mitochondrial dysfunction.

[0405] Therefore, possible synergistic effects between EBS and said compounds were tested in different models:

[0406] - In the taz mutant yeast model

[0407] - Mutants of complex I of the filamentous fungus Podospora (Pa nuo-51 A357V )middle

[0408] - in taz mutant mammalian models.

[0409] Materials and methods

[0410] Taz Yeast:

[0411] As in Figure 1, taz1 mutant yeast strains were spread on respiratory solid medium containing ethanol as a carbon source. The strains were then exposed to filters on which disulfiram (DSF 5nmol), ebselen (EBS 30nmol) or alverine (ALV30nmol) were deposited (alone or a combination of two or three). The plates were then incubated at a temperature of 36°C and then scanned after 5 days of incubation. DMSO (solvent) was used as a negative control (upper left filter).

[0412] Pa nuo-51 A357V :

[0413] Growth rate assessed in cm / day at 32°C was determined in basal medium supplemented with EBS (0.003 μM, 0.01 μM, or 0.03 μM), ALV (1 μM), or a combination of both concentrations, compared to the same concentration of DMSO (VEH) per dish.

[0414] Taz mammalian cells:

[0415] The synergistic effect of EBS, ALV and DSF on the respiratory growth of myoblasts from TAZ patients (carriers of TAZ gene mutations) was tested. Proliferation rates were measured 120 hours after seeding cells with or without individual compounds or in combination at the indicated concentrations in 384-well microtiter plates (at T0, 1200 cells / well) in medium with weak glucose concentrations (to force cells to use the OXPHOS system instead of glycolysis) using a sulforhodamine B (SRB) staining protocol.

[0416] result

[0417] Figure 8 A shows that the combination of the three molecules EBS, ALV and DSF increases the growth halo around the filter where they deposited in a joint manner, indicating a synergistic effect of the three drug candidates on respiratory growth in the tafazzin-deficient yeast model.

[0418] Figure 8 B shows that the combination of 1 μM ALV + 0.003 μM EBS significantly increased the expression of mutant Panuo-51 compared with 1 μM ALV alone. A357V This indicates a synergistic effect on the growth of the complex I mutants.

[0419] Example 7: Mouse Model of Barthel Syndrome

[0420] Materials and methods

[0421] Mice lacking the TAZ gene (KO model of Barthel syndrome) or the wild-type (WT) gene were gavaged twice daily with 10 mg / kg of ebselen starting at day 51. Body weight was monitored regularly during the 1-month treatment period.

[0422] result

[0423] like Fig. 9 As shown in , KO mice treated with EBS appeared to have greater weight compared to mice treated with vehicle.

Claims

1. A pharmaceutical composition comprising a compound having a moiety of the formula: For use in the treatment of diseases associated with mitochondrial dysfunction.

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

3. The composition for use thereof according to claim 2, wherein: The compound is hexaneselenol or one of its derivatives, such as MAD423.

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

5. The composition for use thereof according to claim 4, wherein The compound is: - 2-phenyl-1,2-benzisoselenazol-3(2H)-one or ebselen of the following formula: or - 1-oxide-2-phenyl-1,2-benzisoselenazol-3(2H)-one or ebselenoxide or ebselenoselenoxide of the following formula:

6. The composition for use according to any one of claims 1 to 5, wherein The disease is a disease of the mitochondrial respiratory chain, advantageously a disease of the mitochondrial respiratory chain associated with a Complex I defect.

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

8. The composition for use thereof according to claim 7, wherein 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 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, NDUFV 3. NDUFAF1, NDUFAF2, NDUFAF3, NDUFAF4, NDUFAF5, NDUFAF6, NDUFAF7, NDUFAF8, NUBPL, ACAD9, TMEM70, TMEM126B, FOXRED1, ECSIT, AIF, TIMMDC1.

9. The composition for use thereof according to claim 7, wherein The genetic disease comprises at least one mutation 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.

10. The composition for use according to any one of claims 1 to 9, wherein The disease is selected from the group consisting of: MELAS syndrome, myopathy or cardiomyopathy inherited from the mother, NARP or MILS syndrome, Leigh syndrome, Leber hereditary optic neuropathy (LHON), Barthel syndrome, mitochondrial DNA depletion syndrome, especially 4A (Alpers type) and 4B (MNGIE type), mitochondrial recessive ataxia syndrome, sensory ataxia neuropathy, dysarthria and ophthalmoplegia, spinocerebellar ataxia with epilepsy, progressive external ophthalmoplegia, mitochondrial DNA-6 depletion syndrome, Navajo type neuropathy, Behr syndrome, mitochondrial DNA 14 depletion syndrome, infantile cardioencephalomyopathy due to lack of cytochrome c oxidase (COA6 mutation), mitochondrial type 1 complex III nuclear defect, GRACILE syndrome and Bjornstad syndrome.

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

12. The composition for use thereof according to any one of claims 1 to 7, wherein The disease is selected 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 blood vessel diseases; - 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 disorder, schizophrenia, bipolar and mood disorders; - Gastrointestinal disorders; - Fatigue disorders such as chronic fatigue syndrome and Gulf War illness; -Musculoskeletal disorders such as fibromyalgia and hypertrophy / atrophy of skeletal muscles; - Muscular dystrophy, - cancer; and - Chronic infection.

13. A composition for use thereof 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. A composition for use thereof according to any one of the preceding claims, wherein The composition is administered orally.

15. A composition for use thereof according to any one of the preceding claims, wherein The composition is in solid form such as a tablet, and more advantageously comprises 200 mg of ebselen.

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