Sterol-derived compounds for treatment of diseases associated with mitochondrial deficiency

Abstract

The present invention relates to a compound derived from a sterol of formula (I) for its use in the prevention, amelioration and / or treatment of diseases associated with mitochondrial defects, the disease is selected from the group consisting of autism, spinal cord lesions, multiple sclerosis, epilepsy, migraine, mental disorders associated with drinking, ataxia, neuropathy, brain and lung disorders associated with smoking, MERRF syndrome, and NARP syndrome. The invention also relates to a pharmaceutical composition comprising at least one compound of formula (I).

Description

Technical Field

[0001] The present invention relates to the field of mitochondrial defect-related conditions. More specifically, the present invention relates to a sterol compound of formula (I) and a pharmaceutical composition comprising a sterol compound of formula (I) or a pharmaceutically acceptable salt of such a compound for use in preventing, improving and / or treating mitochondrial defect-related conditions, wherein the mitochondrial defect-related conditions are selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychiatric conditions, ataxia, neuropathy, smoking-related brain and lung diseases, MERRF syndrome and NARP syndrome. Background Art

[0002] Mitochondria are essential organelles present in most eukaryotic cells. Their main role is to provide cells with the energy they need to ensure their survival and the functions they are expected to perform by converting oxygen and food into energy sources such as adenosine triphosphate (ATP). Given this fundamental role played by mitochondria within cells, many pathologies are associated with mitochondrial dysfunction, mainly affecting systems that require a lot of energy, such as muscles, brain or liver.

[0003] Defective or damaged mitochondria will also produce energy in a defective way. There are so-called primary (inherited) and secondary (acquired) mitochondrial diseases. In both cases, the mitochondria no longer function properly and will give rise to various pathologies. These pathologies are related to mitochondrial activity, mitochondrial dynamics (biogenesis, fusion, fission) and axonal transport. Mitochondrial diseases or mitochondropathies combine a range of different diseases related to disorders of the mitochondrial respiratory chain.

[0004] There are a variety of pathologies in which defects in mitochondrial activity and / or mitochondrial dynamics and / or axonal transport are observed, notably autism, spinal cord injury, multiple sclerosis, epilepsy and even migraine (Cleveland Clinic, Mitochondrial diseases, https: / / my.clevelandclinic.org / health / diseases / 15612-mitochondrial-diseases).

[0005] In addition to natural methods such as exercise or adopting a healthy lifestyle (avoiding alcohol, lack of sleep, drastic dieting, etc.), there are currently few molecules that can generally improve mitochondrial function. Coenzyme Q10, vitamin B (thiamine, riboflavin), alpha-lipoic acid, L-carnitine, creatine, L-arginine, idebenone, KH176, Elamipretide, Bezafibrate, resveratrol, omaveloxolone, rapamycin (patent US10792 287) and nicotinamide adenine dinucleotide (NAD+) precursors are often mentioned as potential therapeutic agents for mitochondrial diseases, but none of them has completed all clinical stages. Currently, only vitamin B12 can be administered (Treatment for mitochondrial diseases, Tongling Liufu et al., September 9, 2020, https: / / doi.org / 10.1515 / revneuro-2020-0034; MolecularGenetics and Metabolism,vol.131,September-October 2020,pages 1-13,Clinicaltrials in mitochondrial disorders,an update,Mohammed Almannai et al .,doi: 10.1016 / j.ymgme.2020.10.002).

[0006] Therefore, there are currently no FDA-approved therapies for the treatment of mitochondrial dysfunction (Trends in Molecular Medicine, Special issue: Mitochondria–from diagnosis to treatment review, vol. 26, issue 1, pages 40-57, January 01, 2020 , https: / / doi.org / 10.1016 / j.molmed.2019.09.002 ).

[0007] Mitochondrial diseases are primarily the result of mutations or deletions in genes encoding mitochondrial proteins. Thus, mitochondrial function can be impaired due to inherited mutations or acquired or iatrogenic mitochondrial damage. Improving or restoring mitochondrial function (when it is defective) is therefore a fundamental challenge to combat all of these pathologies or at least some of the consequences identified as mitochondrial defects.

[0008] Autism spectrum disorder is a neurodevelopmental disease that causes social behavior deficits and cognitive impairments. Currently, mitochondrial dysfunction is considered a possible cause of this disorder, especially defects in mitochondrial dynamics (mitochondrial fission), mitochondrial biogenesis, or axonal transport of these mitochondria (Seminar in Pediatric Neurology, vol. 25, October 2020, Richard E. Frye et al., Mitochondrial dysfunction in autism spectrum disorder: unique abnormalities and targeted treatments, https: / / doi.org / 10.1016 / j.spen.2020.100829).

[0009] During spinal cord injury, axons degenerate. Increasing evidence suggests that mitochondria play a fundamental role in the degeneration / regeneration of these axons, both in terms of mitochondrial activity, dynamics, and mitochondrial axonal transport (Frontiers in aging neuroscience, March 8, 2021, Biyao Wang et al., Mitochondrial behavior in axon degeneration and regeneration, https: / / doi.org / 10.3389 / fnagi.2021.650038).

[0010] Multiple sclerosis is a disease in which demyelination of nerves leads to axonal damage and various neurological symptoms. Defects in mitochondrial activity and transport have also been observed in this condition (Immunology and Inflammation, Neuroscience, February 10, 2021, Sina C Rosenkranz et al., Enhancing mitochondrial activity in neurons protects against neurodegeneration in a mouse model of multiple sclerosis, https: / / elifesciences.org / articles / 61798).

[0011] Mitochondrial dysfunction is also considered to be a potential cause of epilepsy (European Journal of Pediatric Neurology, vol. 24, pages 47-52, January 01, 2020, Albert Lim et al., The mitochondrial epilepsies, https: / / www.ejpn-journal.com / article / S1090-3798(19)30441-6 / fulltext). Antioxidants targeting mitochondrial oxidative stress have also been shown to be promising neuroprotectants for epilepsy (Oxidative Medicine and Cellular Longevity, vol. 2020, Article ID 6687185, Nan Yang et al., Antioxidants targeting mitochondrial oxidative stress: promising neuroprotectants for epilpesy, https: / / doi.org / 10.1155 / 2020 / 6687185). Recurrent migraine attacks are often associated with metabolic changes in specific brain regions. These metabolic changes arise from defects in mitochondrial function in these areas (Headache: The Journal of Head and Face Pain, 2018, vol. 58: pages 45-52, Kraya T. et al., Prevalence of headache in patients with mitochondrial disease: a cross-sectional study, https: / / americanheadachesociety.org / news / journal-headache-mitochondrial-disease / ).

[0012] Mitochondrial dysfunction has been described in alcohol-related psychotic symptoms. Various psychotic symptoms may be associated with drinking, such as independent major depression, bipolar disorder, anxiety or personality disorder (Shivani, R., Goldsmith, RJ, & Anthenelli, RM (2002). Alcoholism and Psychiatric Disorders: Diagnostic Challenges. Alcohol Research & Health, 26 (2), 90–98). The mechanism of ethanol toxicity in brain neurons has been described, in which mitochondria are the main mediators, and mitochondrial changes are associated with the severity of drinking. Therefore, improving the health of brain cell mitochondria is being considered as a potential therapeutic target for treating ethanol-related pathologies (Ethanol Consumption Affects Neuronal Function: Role of the Mitochondria, Cheril Tapia-Rojas et al., December 20, 2017, DOI: 10.5772 / intechopen.71611). It has also been shown that adolescent drinking has a significant impact on mitochondrial bioenergetics in adulthood, and this effect is not a temporary change before drinking stops, as previously thought (Neuroscience, vol. 406, May 15, 2019, pages 356-368, Cheril Tapia-Rojas et al., Adolescence binge alcohol consumption induces hippocampal mitochondrial impairment that persists during the adulthood).

[0013] Ataxia is a neuromuscular disorder consisting of a deficit in the fine coordination of voluntary movements. It is associated with damage to the nervous system. Ataxias associated with mitochondrial defects include Friedreich's syndrome (Molecular and Cellular Neuroscience, vol. 102, January (2020), Anna Stepanova et al., Mitochondrial dysfunction in neurons in Friedreich's ataxia, https: / / www.sciencedirect.com / science / article / abs / pii / S1044743119301964?via%3Dihub), telangiectasias (Scientific Report 9, 4782 (2019), Blignaut, M. et al., Ataxia-Telangiectasia Mutated is located in cardiac mitochondria and impacts oxidative phosphorylation, https: / / www.nature.com / articles / s41598-019-41108-1); spinocerebellar ataxia (Cell Reports, vol. 26, issue 5, pages 1189-1202, January 29, 2019, Metabolic and organelle morphology defects in mice and human patients define spinocerebellar ataxia type 7 as a mitochondrial disease, https: / / www.cell.com / cell-reports / fulltext / S22111247(19)300373? _returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS22111247193

[0014] 00373%3Fshowall%3Dtrue#relatedArticles).

[0015] Neuropathy and myopathy have also been shown to be associated with mitochondrial dysfunction (Mitochondrion, 56 (2021), 52-61, Jian-Qiang Lu et al., Mitochondrial neuropathy and neurogenic features inmitochondrial myopath., https: / / doi.org / 10.1016 / j.mito.2020.11.005).

[0016] Many publications have confirmed the brain and lung diseases related to smoking, in which nicotine or tobacco smoke has an effect on various mitochondrial activities. Mitochondria have been presented as possible targets for nicotine in the brain (Journal of Bioenergetics and Biomembranes, 51, 259-276 (2019), Dominika Malinska et al., Mitochondria as a possible target for nicotine action), and the disorder of mitochondrial function and structure of lung epithelial cells has been shown to be the effect of cigarette smoke (American Journal of Physiology, vol. 318, No. 1, January 7, 2020, Mathyar Aghapour et al., Mitochondria: at the crossroads of regulating lung epithelial cell function in chronic obstructive pulmonary disease, https: / / doi.org / 10.1152 / ajplung.00329.2019).

[0017] Finally, MERRF (myoclonic epilepsy with ragged red fibers) syndrome and NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome are both associated with mitochondrial defects (Elsevier, Biochimica et Biphysica Acta (BBA) Molecular Basis of Disease, vol. 1866, issue 6, June 1, 2020, Marina Vilanueva-Paz et al., Parkin-mediated mitophagy and autophagy flux disruption in cellular models of MERRF syndrome; Elsevier, The International Journal of Biochemistry & Cell Biology, vol. 45, issue A, January 2013, pages 141-150, Magdanela Lebiedzinska et al., Disrupted ATP synthase activity and mitochondrial hyperpolarization-dependent oxidative stress is associated with p66Shcphosphorylation in fibroblasts of NAR patients).

[0018] US 10 792 287 describes the use of pharmaceutical compounds (zolpidene with the addition of rapamycin or idebedone) for their beneficial effects on mitochondrial ATP production, inhibition of inflammation and for the treatment of various mitochondrial pathologies such as vision loss and Leber's hereditary optic neuropathy.

[0019] US 10 272 056 also describes the use of butyrate-type compounds for their beneficial effects on mitochondrial biogenesis, increasing mitochondrial mass, increasing ATP production, and their therapeutic efficacy in various pathologies such as neurodegenerative diseases, cardiovascular, neurometabolic, muscle, renal and metabolic diseases, as well as certain specific syndromes such as MELAS (mitochondrial encephalopathy with lactic acidosis and stroke-like episodes) and MERRF syndrome.

[0020] The applicant surprisingly found that the sterol compound of formula (I) or a pharmaceutically acceptable salt of such a compound can be used to prevent, improve and / or treat mitochondrial defect-related conditions caused by defects in mitochondrial dynamics and / or transport and / or activity. More specifically, the applicant found that the sterol compound of formula (I) or a pharmaceutically acceptable salt of such a compound can be used to prevent, improve and / or treat mitochondrial defect-related conditions selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychotic symptoms, ataxia, neuropathy, smoking-related brain and lung diseases, MERRF syndrome and NARP syndrome. Summary of the invention

[0021] The object of the present invention is to provide sterol-based compounds and pharmaceutical compositions comprising the same for use in preventing, ameliorating and / or treating diseases associated with mitochondrial defects, wherein the diseases associated with mitochondrial defects are selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychotic symptoms, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome and NARP syndrome.

[0022] To this end, the present invention provides a compound of formula (I) for use in treating conditions associated with mitochondrial defects:

[0023]

[0024] in

[0025] R1=OH、F、OC n H 2n+1 , OC(O)R, OC(O)OR, OC(O)NHR or OP(O)(OR)2,

[0026] Where R=H or C n H 2n+1 , where 1≤n≤8;

[0027] R2=H or OH;

[0028] R3=–NR5R6,

[0029] R5 is H or –(CH2)3NH2, and

[0030] R6 is included in the group formed by: –(CH2)3NR7(CH2)4NHR7, –(CH2)3NHR7, –(CH2)4NHR7, –(CH2)4NR7(CH2)3NHR7, –(CH2)3NR7(CH2)4NR7(CH2)3NHR7, –(CH2)2-imidazol-4-yl, –(CH2)2-indol-3-yl, wherein R7=H, C(O)OCH3 or C(O)OC(CH3)3; and

[0031] R4 = H or OH, at position 20, 22, 24, 25, 26 or 27, positioned to produce an asymmetric center of R or S configuration;

[0032] Z1 and Z2 each represent the number of double bonds (0 or 1) between carbon atoms C7 and C8 and C22 and C23, respectively; T1, T2 and T3 are independently of each other = H or CH3; T4 = H, CH3, C2H5, positioned to obtain an asymmetric center of R or S configuration at position 24;

[0033] The mitochondrial defect-related condition is selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychiatric conditions, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome, and NARP syndrome.

[0034] The mention of (O) in the definition of radicals according to formula (I) means that the oxygen is bound via two bonds.

[0035] The compound of formula (I) is defined as follows: Z1=Z2=0; R1=R2=OH; R4=H; R5=H; R6=-(CH2)3-NC(O)OC(CH3)3-(CH2)4-NHC(O)OC(CH3) 3； T1=T2=T3=T4=H, referred to as DX243BOC or DXboc, and is shown in Table 2.

[0036] The substituent or group C(O)OC(CH3)3 is also known as tert-butoxycarbonyl or Boc functionality.

[0037] The compound of formula (I) belongs to the steroid group. Therefore, the numbering of the carbon atoms of the compound of formula (I) follows the nomenclature defined by IUPAC in Pure & Appl. Chem., Vol. 61, No. 10, pages 1783-1822, 1989. The numbering of the carbon atoms of the compound belonging to the steroid group according to IUPAC is as follows:

[0038]

[0039] Processes for the preparation of compounds of formula (I) have been described previously, in particular in de Medina, P. et al. Synthesis of New, the Treatment of Cancer and Neurodegenerative Diseases. Journal of Medicinal Chemistry, 52(23), 2009, pp. 7765-7777.

[0040] In addition, the compound may have one or more of the following characteristics considered alone or in combination. According to one embodiment, the compound of formula (I) is defined as follows: R1 = OH, F, OC n H 2n+1 , OC(O)R, OC(O)OR, OC(O)NHR or OP(O)(OR)2, where R=H or C n H 2n+1 , where 1≤n≤8;

[0041] R2=OH;

[0042] R3=–NR5R6;

[0043] R5=H;

[0044] R6=–(CH2)3NR7(CH2)4NHR7, –(CH2)3NHR7, –(CH2)4NHR7, –(CH2)4N

[0045] R7(CH2)3NHR7, –(CH2)3N R7(CH2)4N R7(CH2)3NHR7, –(CH2)2-imidazol-4-yl, –(CH2)2-indol-3-yl, wherein R7=H, C(O)OCH3 or C(O)OC(CH3)3;

[0046] Z1=0 or 1;

[0047] Z2=0;

[0048] R4=H.

[0049] According to one embodiment, the compound of formula (I) is more precisely defined as follows: Z1=0, Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H, wherein the other groups R3, R6 and R7 are as defined above.

[0050] According to one embodiment, the compound of formula (I) is more precisely defined as follows: Z1=0; Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H, wherein R6=-(CH2)4NH(CH2)3NHR7 and R7=C(O)CH3. This compound named DX249 is 5α-hydroxy-6β-[3-(4-aminobutylacetamide)propylamino]cholestan-3β-ol.

[0051] According to one embodiment, the compound of formula (I) is more precisely defined as follows: Z1=0; Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H, wherein R6=-(CH2)2-imidazol-4-yl. This compound named DX101 is 5α-hydroxy-6β-[2-(1H-imidazol-4-yl)ethylamino]cholestan-3β-ol.

[0052] According to one embodiment, the compound of formula (I) is more precisely defined as follows: Z1=0; Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H, wherein R6=-(CH2)3NR7(CH2)4NHR7, -(CH2)4NR7(CH2)3NHR7, -(CH2)3NR7(CH2)4NH(CH2)3NHR7; or -(CH2)4NHR7; and R7=H. These compounds, respectively named DX243, DX245, DX301 and DX401, are:

[0053] -5α-Hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholestan-3β-ol (DX243)

[0054] -5α-Hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholestan-3β-ol (DX245)

[0055] -5α-Hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholestane-3β-ol (DX301)

[0056] -5α-Hydroxy-6β-(4-aminobutylamino)cholestane-3β-ol (DX401).

[0057] According to one embodiment, the compound of formula (I) is more precisely defined as follows: Z1=0, Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H, wherein R6=-(CH2)3NR7(CH2)4NHR7 and R7=C(O)OC(CH3)3. This compound named DX243BOC is 5α-hydroxy-6β-[3-(4-tert-butoxycarbonylaminobutyl-tert-butoxycarbonylamino)propylamino]cholestan-3β-ol.

[0058] According to one embodiment, the compound of formula (I) is more precisely defined as follows: Z1=1, Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H, wherein the other groups R3, R6 and R7 are as defined above.

[0059] According to one embodiment, the compound of formula (I) is more precisely defined as follows: Z1=1; Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H, wherein R6=-(CH2)3NH(CH2)4NHR7;-(CH2)4NH(CH2)3NHR7; or-(CH2)3NH(CH2)4NH(CH2)3NHR7; and R7=H. These compounds, respectively named DX242, DX244 and DX302, are:

[0060] -5α-Hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholester-7-en-3β-ol (DX242)

[0061] -5α-Hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholester-7-en-3β-ol (DX244)

[0062] -5α-Hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholester-7-en-3β-ol (DX302).

[0063] According to one embodiment, the pathology associated with mitochondrial defects is due to defects in mitochondrial dynamics and / or trafficking and / or activity. Table 1 illustrates the specificity of the defects according to the pathology.

[0064] Table 1

[0065]

[0066]

[0067] For the compounds of formula (I) according to the invention, the pathologies associated with mitochondrial defects are due to defects in mitochondrial dynamics and / or mitochondrial transport and / or mitochondrial activity.

[0068] In one embodiment, the condition associated with defects in mitochondrial dynamics and mitochondrial trafficking and activity is a neurological disorder, spinal cord injury, autism, ataxia, and alcohol-related psychotic conditions.

[0069] In another embodiment, the condition associated with defects in mitochondrial dynamics and mitochondrial activity is migraine, smoking-related brain and lung disorders, and MERRF syndrome.

[0070] In another embodiment, the condition associated with defects in mitochondrial trafficking and activity is multiple sclerosis.

[0071] In yet another embodiment, the condition associated with defects in mitochondrial activity is NARP syndrome.

[0072] A second subject matter according to the present invention is a pharmaceutical composition comprising at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt of at least one compound of formula (I) for use in the treatment of mitochondrial defect-related conditions selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychotic conditions, ataxia, neuropathies, smoking-related brain and lung diseases, MERRF syndrome and NARP syndrome.

[0073] The composition for use according to the invention can be administered in various forms suitable for the condition to be treated. Thus, various administration forms include oral, topical, systemic, intravenous, subcutaneous, intraperitoneal, intramuscular, transdermal or transmucosal administration.

[0074] The composition according to the invention may be in any presentation form normally used, depending on whether the composition is to be ingested, injected or applied to the skin or mucous membranes.

[0075] The composition according to the invention may include ingredients commonly used in preparations of this type, such as binders, flavoring agents, preservatives or coloring agents, and in the case of food supplements or medicaments, may be in the form of tablets, granules or gel capsules. The preparation according to the invention may also be in the form of a food, such as a beverage, or in the form of a suspension or syrup.

[0076] According to a first variant, the various formulations are suitable for topical administration and include creams, oil-in-water and water-in-oil emulsions, milks, ointments, lotions, oils, balms, aqueous or aqueous-alcoholic or ethanolic solutions, slurries, powders, patches, sprays or any other product for external application, such as a medical device or an aerosol product also comprising a pressurized propellant.

[0077] According to a second variant, the various compositions are suitable for injection; the compositions may be in the form of an aqueous or oily agent, or in the form of a serum.

[0078] According to a third variant, the various compositions are suitable for ingestion; the composition may be in the form of capsules, syrups, granules or tablets.

[0079] According to preferred embodiments, compositions according to the present invention are more particularly intended for topical administration. Therefore, these compositions must comprise dermatologically acceptable media, i.e. media compatible with skin and mucous membranes, and encompass all dermatological forms. These compositions can be in the following form in particular: emulsifiable paste, oil-in-water or water-in-oil emulsion or composite emulsion, serosity, solution, suspension, gel, milk, lotion, stick or powder, and can be applicable to being applied to skin and mucous membranes. These compositions include the necessary excipients for their preparations, such as solvents, emollients, thickeners, diluents, surfactants, antioxidants, bioactivators, dyes, preservatives and spices.

[0080] The compositions according to the invention also comprise any additives usually used in the envisaged field of application and also the adjuvants required for their formulation, such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens, self-tanning agents, pigments, fillers, preservatives, fragrances, odor absorbers, dermatologically or pharmaceutically active agents, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0081] In any case, the skilled person will take care to ensure that these adjuvants and their proportions are chosen so that the desired beneficial properties of the composition according to the invention are not adversely affected.

[0082] According to one embodiment, the composition for use according to the present invention is in the form of an aqueous solution and has a concentration of 1 pmol.L -1 To 1mmol.L -1 , preferably 10pmol.L -1 To 0.1mmol.L -1 , more preferably 0.1nmol.L -1 To 1μmol.L -1 The concentration of the compound of formula (I) is BRIEF DESCRIPTION OF THE DRAWINGS

[0083] The invention will be better understood and other objects, details, features and advantages thereof will appear more clearly from the following description of several specific embodiments of the invention with reference to the accompanying drawings, which description is given as a non-limiting illustration only.

[0084] [ Figure 1 ] Figure 1 The results of a test of the activity of compounds of formula (I) on cortical neurons by measuring the optical density of the tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) are illustrated.

[0085] NT means no treatment (untreated). The results allow identification of the relative metabolic activity of cortical neurons with or without the compound of formula (I).

[0086] [ Figure 2A ] Figure 2A The results of the test of the activity of Neuro-2a cells under normal conditions (without deprivation of oxygen and glucose) using compounds of formula (I) are described by measuring the optical density of the tetrazolium salt MTT. CTL means no treatment (control). The results allow the identification of the relative metabolic activity of Neuro-2a cells with or without compounds of formula (I).

[0087] [ Figure 2B ] Figure 2B The beneficial effects of compounds according to formula (I) on the metabolic activity of Neuro-2a cells after ethanol (EtOH) intoxication are described and demonstrated. Figure 2C ] Figure 2C The beneficial effects of compounds according to formula (I) on mitochondrial activity of Neuro-2a cells after ethanol (EtOH) intoxication are described and demonstrated.

[0088] [ Figure 3 ] Figure 3 The results of tests demonstrating the effect of compounds of formula (I) on mitochondrial dynamics in cell lines are described.

[0089] [ Figure 4 ] Figure 4 The results of tests demonstrating the inhibition of mitochondrial fission with compounds according to formula (I) are illustrated. qRT-PCR analysis of cochlear explants treated with compound DX243 enabled the observation of a reduction in drp1 mRNA after 24h of treatment with 100 nM DX243 relative to non-treated (NT) cochleae.

[0090] [ Figure 5 ] Figure 5 Results of tests assessing mitochondrial dynamics and mitochondrial stress in dopaminergic neurons are described.

[0091] [ Figure 6 ] Figure 6 Described are the results of tests assessing mitochondrial stress in cultures of dopaminergic neurons.

[0092] [ Figure 7 ] Figure 7 Results of tests assessing mitochondrial stress in cultures of hippocampal neurons are illustrated.

[0093] exist Figure 2A , 2C , 3, 5 and 6, the asterisks indicate the statistical power of the results. One asterisk indicates 95% confidence that the result is not due to chance. The presence of two asterisks indicates 99% confidence that the result is not due to chance, the presence of three asterisks indicates 99.9% confidence that the result is not due to chance, and the presence of four asterisks indicates 99.99% confidence that the result is not due to chance. DETAILED DESCRIPTION

[0094] In this description, unless otherwise stated, it is understood that when a range is given, it includes the upper and lower limits of the range.

[0095] In the present invention, the following definitions apply:

[0096] "Preventing a condition" means avoiding the occurrence of disease or injury, or maintaining and improving health; it may also be referred to as preventive treatment.

[0097] "Ameliorating a condition" means alleviating the symptoms of the condition; it may also be called palliative treatment.

[0098] "Treating a condition" means curing the disease; it may also be referred to as curative treatment.

[0099] "Mitochondrial defect" means a reduction in mitochondrial activity and / or mitochondrial dynamics and / or mitochondrial axonal transport, leading to loss of energy production and accumulation of substances that are harmful to the body.

[0100] "Mitochondrial activity" refers to the ability of mitochondria to regulate cellular metabolism and produce energy (ATP) through their respiratory chain.

[0101] "Mitochondrial dynamics" refers to the ability of mitochondria to fuse or fission in order to maintain their morphology and size and / or allow for an increase in the amount (mass) of mitochondria in the cell.

[0102] "Mitochondrial axonal transport" refers to the ability of mitochondria to move to meet necessary energy demands at precise points in the cell.

[0103] "Mitochondrial fission" means the splitting of a mitochondria into two distinct mitochondria.

[0104] In order to prove that the compound of formula (I) has an impact on the mitochondrial function of cells, various experiments have been carried out, demonstrating that the compound generally has a beneficial effect on mitochondrial function by its effect on the number, dynamics, stress, transport or activity of mitochondria. Several experimental schemes are described below, which demonstrate the beneficial effects of the compound of formula (I) (particularly those shown in Table 2 below) under different conditions and experimental models.

[0105] Table 2: List of compounds of formula I used in the experimental examples.

[0106]

[0107] The concentration or molarity of a compound is expressed in moles per liter, and its symbol is mol.L -1 or M.

[0108] Example 1: Testing of relative metabolic activity of cortical neurons

[0109] A protocol was developed for obtaining primary cultures of cortical neurons from cells collected from the brains of wild-type mouse embryos. Step 1 involves collecting the embryonic cortex to place the cortical neurons under appropriate culture conditions. Step 2 involves culturing the cortical neurons in Neurobasal TM The collected cells were cultured in culture medium (reference number 21103049 ThermoFisher Scientific) to which L-glutamine and B27 supplement 50X (reference number 17504044 ThermoFisher Scientific) were added. The neurons of the primary culture were then isolated and purified. The above culture conditions provide purified neuronal cultures from the dissociation of embryonic cortices.

[0110] A test based on the metabolic activity of neurons was performed to assess the effect of compounds of formula (I) on this metabolic activity. The test is based on the use of the tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). Tetrazolium is reduced to a purple precipitate formazan by mitochondrial succinate dehydrogenase in active living cells. The amount of precipitate formed is proportional to the metabolic activity present in the culture. Therefore, the spectroscopic determination of the optical density at 550nm makes it possible to determine the relative amount of metabolic activity and therefore mitochondrial activity. The results obtained are shown in Figure 1 This test on cortical neuron cultures was performed with the different compounds mentioned in Table 2 (DX101, DX243, DX244, DX245, DX249, DX301, DX302 and DX401) used at the same concentration (100 nM).

[0111] refer to Figure 1, which test makes it possible to highlight the positive effects of the test compounds of formula (I) on mitochondrial activity relative to untreated neurons.

[0112] Example 2: Testing of relative metabolic activity of Neuro-2a neurons

[0113] The same tests described in Example 1 were carried out on Neuro-2a cells (mouse neuroblastoma cell line). Once cultured and at a confluence of about 50%, Neuro-2a cells were placed in contact with two concentrations of compounds DX243 and DX245 for 24 h under physiological conditions. The MTT assay carried out after these 24 h treatments made it possible to demonstrate that compound DX243, but especially compound DX245, allowed to increase the metabolic activity of Neuro-2a cells, such as Figure 2A What is presented.

[0114] Example 3: Testing of metabolic activity after ethanol (EtOH) intoxication

[0115] The same test as described in Example 1 was performed on Neuro-2a cells (mouse neuroblastoma cell line). It was shown that intoxication with high concentration (500 mM) of ethanol (EtOH) for 24 h in this cell line reduced the metabolic activity of these cells (χ=P value <0.0001, relative to control). This metabolic activity was partially restored by treatment with 1 nM compound DX243 for 24 h.

[0116] Interestingly, after intoxication with EtOH (200 mM and 500 mM), treatment with 100 nM of compound DX243 or compound DX245 for 24 h resulted in significantly greater metabolic activity compared to untreated cells, as shown in Figure 2 Figure 2B shown.

[0117] Figure 2C The effect of the compound DX243 on mitochondria in Neuro-2a cells after ethanol intoxication is shown. The figure highlights the increase in mitochondrial surface area in the presence of the compound DX243, which was studied with the help of MitoTracker after 24h of ethanol intoxication (500 mm).

[0118] - Example 4: Testing of mitochondrial dynamics in SHSY5Y and C2C12 cell lines

[0119] 5. 5. 4. 5. 2. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. TM Human neuroblastoma cell line (SHSY5y) and myoblast cell line (C2C12) were cultured in culture medium (reference number 21068028 ThermoFisher Scientific). Once in culture, both cell lines were treated with different concentrations of compound DX243 for 24 h. A series of images were acquired and the expression of the compound was detected using Mitotracker TM(reference number M7512, ThermoFisherScientific), these images highlight and visualize the mitochondria present in these cells and enable quantification of their amount. Quantification of mitochondrial mass revealed by Mitotracker labeling highlights the effect of compound DX243 on mitochondrial dynamics (increase in mitochondrial amount (mass)), as Figure 3 shown.

[0120] Example 5: Assays to assess inhibition of mitochondrial fission in cochlear explants

[0121] A cochlear explant model was developed. In order to obtain the cultured auditory organ cochlea, the cochlea of ​​3-day-old mice was microdissected. These cochleae were then placed in DMEM medium supplemented with glucose (0.5%), N1 (0.5%), insulin (0.25%) and penicillin (0.1%) for 24 hours. Then 100nM compound DX243 was added to the medium for 24 hours.

[0122] The cochlea was then recovered and RNA was extracted from it, allowing analysis of the regulation of several genes after treatment with the compound DX243. Thus, it was observed that treatment of cochlear explants with the compound DX243 (100 nM) induced an inhibition of the expression of the mitochondrial fission gene drp1, as Figure 4 This test demonstrated the beneficial effects of compound DX243 on mitochondria, specifically by inhibiting mitochondrial fission.

[0123] Example 6: Assays to assess mitochondrial dynamics and mitochondrial stress in dopaminergic neurons

[0124] The mitochondrial effects of compound DX243 were tested in a model of dopaminergic neurons treated with MPP+ (1-methyl-4-phenylpyridinium), a mitochondrial toxin that increases mitochondrial stress and reduces mitochondrial number. The deleterious effects of MPP+ on mitochondria are well known (increased mitochondrial stress measured by the amount of cytochrome C, reduced mitochondrial number measured by Mitotracker assay). Dopaminergic neurons were isolated from mice treated with Neurobasal TMObtained from microdissection of the midbrain of rat embryos cultured in medium (reference number 21103049 ThermoFisher Scientific) supplemented with B27 supplement 50X (reference number 17504044 ThermoFisher Scientific), penicillin / streptomycin, L-glutamine and also BDNF (brain-derived neurotrophic factor) and GDNF (glial cell-derived neurotrophic factor). The neurons were cultured for 5 days and then treated with MPP+ for 48h. The neurons were also exposed to different concentrations of compound DX243 during the 48h of MPP+ treatment and also 48h after MPP+ removal.

[0125] The results demonstrated that the compound DX243 was able to increase and restore mitochondrial mass in these dopaminergic neurons that had lost some of this mass after MPP+ treatment, e.g. Figure 5 shown.

[0126] In addition, 10 nM of compound DX243 also reduced mitochondrial stress caused by MPP+ treatment, such as Figure 6 shown.

[0127] Example 7: Assays to assess mitochondrial dynamics and mitochondrial stress in hippocampal neurons

[0128] The mitochondrial effects of compound DX243 were tested in a model of hippocampal neurons treated with the peptide Aβ1-42, which induces mitochondrial stress (represented by a decrease in mitochondrial surface area). Figure 7 As shown, it was shown that compound DX243 (1 nM) was able to restore this reduction in mitochondrial surface area.

[0129] Use of the verb "to contain", "to comprise" or "to include" and its conjugations does not exclude the presence of elements or steps other than those stated in a claim.

[0130] In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim.

Claims

1. A compound of formula (I) for use in treating conditions associated with mitochondrial defects: in R1 = OH, F, OC n H 2n+1 , OC(O)R, OC(O)OR, OC(O)NHR or OP(O)(OR)2, Where R=H or C n H 2n+1 , where 1≤n≤8; R2=H or OH; R3=–NR5R6; R5 is H or –(CH2)3NH2; and R6 is included in the group formed by: –(CH2)3NR7(CH2)4NHR7, –(CH2)3NHR7, –(CH2)4NHR7, –(CH2)4NR7(CH2)3NHR7, –(CH2)3NR7(CH2)4N R7(CH2)3NHR7, –(CH2)2-imidazol-4-yl, –(CH2)2-indol-3-yl, wherein R7=H, C(O)OCH3 or C(O)OC(CH3)3; and R4 = H or OH, at position 20, 22, 24, 25, 26 or 27, positioned to produce an asymmetric center of R or S configuration; Z1 and Z2 each represent the number of double bonds (0 or 1) between carbon atoms C7 and C8 and C22 and C23, respectively; T1, T2 and T3 are independently of each other = H or CH3; T4 = H, CH3, C2H5, positioned to obtain an asymmetric center of R or S configuration at position 24; The mitochondrial defect-related condition is selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychiatric conditions, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome, and NARP syndrome.

2. The compound for use according to claim 1, wherein The compound of formula (I) is defined as follows: R1 = OH, F, OC n H 2n+1 , OC(O)R, OC(O)OR, OC(O)NHR or OP(O)(OR)2, Where R=H or C n H 2n+1 , where 1≤n≤8; R2=OH; R3=–NR5R6; R5=H; R6=–(CH2)3NR7(CH2)4NHR7, –(CH2)3NHR7, –(CH2)4NHR7, –(CH2)4N R7(CH2)3NHR7, –(CH2)3N R7(CH2)4N R7(CH2)3NHR7, –(CH2)2-imidazol-4-yl, –(CH2)2-indol-3-yl, wherein R7=H, C(O)OCH3 or C(O)OC(CH3)3; Z1=0 or 1; Z2＝0； R4=H.

3. The compound for use according to claim 1 or 2, wherein The compound of formula (I) is defined as follows: Z1=0, Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H.

4. The compound for use according to claim 3, wherein The compound of formula (I) is defined as follows: R6 = -(CH2)4NH(CH2)3NHR7, wherein R7 = C(O)CH3, and is the compound 5α-hydroxy-6β-[3-(4-aminobutylacetamide)propylamino]cholestan-3β-ol.

5. The compound for use according to claim 3, wherein The compound of formula (I) is defined as follows: R6 = -(CH2)2NHR7, wherein R7 = imidazol-4-yl, and is 5α-hydroxy-6β-[2-(1H-imidazol-4-yl)ethylamino]cholestan-3β-ol.

6. The compound for use according to claim 3, wherein The compound of formula (I) is defined as follows: R6=-(CH2)3NR7(CH2)4NHR7, -(CH2)4NR7(CH2)3NHR7, -(CH2)3NR7(CH2)4NR7(CH2)3NHR7; or -(CH2)4NHR7; and R7=H, and are 5α-hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholestane-3β-ol, 5α-hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholestane-3β-ol, 5α-hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholestane-3β-ol, 5α-hydroxy-6β-(4-aminobutylamino)cholestane-3β-ol, respectively.

7. The compound for use according to claim 3, wherein The compound of formula (I) is defined as follows: R6 = -(CH2)3NR7(CH2)4NHR7 and R7 = C(O)OC(CH3)3, and is 5α-hydroxy-6β-[3-(4-tert-butoxycarbonylaminobutyl-tert-butoxycarbonylamino)propylamino]cholestan-3β-ol.

8. The compound for use according to claim 2, wherein The compound of formula (I) is defined as follows: Z1=1 and Z2=0; R1=R2=OH; R4=H; R5=H; T1=T2=T3=T4=H.

9. The compound for use according to claim 8, wherein The compound of formula (I) is defined as follows: R6=-(CH2)3NR7(CH2)4NHR7; -(CH2)4NR7(CH2)3NHR7 or -(CH2)3NR7(CH2)4NR7(CH2)3NHR7; and R7=H, and are 5α-hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholester-7-en-3β-ol, 5α-hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholester-7-en-3β-ol, 5α-hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholester-7-en-3β-ol, respectively.

10. The compound for use according to any one of claims 1 to 9, wherein Said mitochondrial defect-related pathologies are due to defects in mitochondrial dynamics and / or mitochondrial transport and / or mitochondrial activity.

11. A pharmaceutical composition comprising at least one compound of formula (I) according to any one of claims 1 to 9 and / or at least one pharmaceutically acceptable salt of at least one compound of formula (I) for use in the treatment of a condition associated with mitochondrial defects selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychotic conditions, ataxia, neuropathies, smoking-related brain and lung diseases, MERRF syndrome and NARP syndrome.

12. The composition according to claim 11, characterized in that It is in the form of an aqueous solution and has a concentration of 1 pmol.L -1 To 1mmol.L -1 , preferably 10pmol.L -1 To 0.1mmol.L -1 , more preferably 0.1nmol.L -1 To 1μmol.L -1 The concentration of the compound of formula (I) is

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