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

Abstract

A sterol-based compound of formula (I) for use in the prevention, amelioration, and / or treatment of mitochondrial deficiency-related medical conditions selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychosis, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome, and NARP syndrome. A pharmaceutical composition comprising at least one compound of formula (I).

Description

Technical Field

[0001] The present invention relates to the field of mitochondrial deficiency-related pathologies. More precisely, the present invention relates to a sterol compound of formula (I) for use in the prevention, amelioration and / or treatment of mitochondrial deficiency-related pathologies 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, and a pharmaceutical composition comprising a sterol compound of formula (I) or a pharmaceutically acceptable salt of such a compound.

Background Art

[0002] Mitochondria are basic organelles present in most eukaryotic cells. Its main role is to provide the cell with the energy necessary for cell survival and expected functions by converting oxygen and food into energy sources such as adenosine triphosphate (ATP). Considering this basic role played by mitochondria within the cell, there are numerous pathologies associated with mitochondrial dysfunction, mainly affecting systems that require large amounts of energy such as muscle, brain, and liver.

[0003] Defective or damaged mitochondria also become incomplete in the way they generate energy. There are what are called primary (hereditary) and secondary (acquired) mitochondrial diseases. In both cases, the mitochondria cease to function properly, causing various pathologies. These pathologies are related to mitochondrial activity, mitochondrial dynamics (biosynthesis, fusion, fission), and axonal transport. Mitochondrial diseases, or mitochondropathies, are a combination of various diseases associated with disorders of the mitochondrial respiratory chain.

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

[0005] Except for natural means such as exercise and adopting a healthy lifestyle (avoiding alcohol, lack of sleep, extreme diets, etc.), there are currently very few molecules that can globally 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 US 10 792 287), and nicotinamide adenine dinucleotide (NAD+) precursors are frequently mentioned as potential therapeutic agents for mitochondrial diseases, but not a single molecule has passed all clinical stages. At present, 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 ; Molecular Genetics and Metabolism, vol. 131, September-October 2020, pages 1-13, Clinical trials in mitochondrial disorders, an update, Mohammed Almannai et al ., doi: 10.1016 / j.ymgme.2020.10.002 )。

[0006] Therefore, currently, there is no FDA-approved treatment targeting 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 mainly the result of mutations or deletions in genes encoding mitochondrial proteins. Therefore, mitochondrial function can be impaired by genetic mutations, or by acquired or iatrogenic mitochondrial damage. Therefore, improving or restoring mitochondrial function when it is reduced is a fundamental challenge in combating all of these pathologies, or at least a part of the pathologies identified as being the result of mitochondrial deficiency.

[0008] The autism spectrum is a neurodevelopmental disorder leading to defects in social behavior and cognitive impairment. Mitochondrial dysfunction is currently being considered as a cause of this disorder, particularly 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] In spinal cord injury, axons degenerate. There is increasing evidence that mitochondria play a fundamental role in the degeneration / regeneration process of these axons, from the perspectives 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 axons are damaged due to demyelination of nerves, causing various neurological symptoms. Defects in mitochondrial activity and transport have also been observed in this pathology (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 has also been identified as 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 been shown to be promising as neuroprotective agents 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 regularly associated with metabolic changes in specific brain regions. These metabolic changes are caused by defects in mitochondrial function present in these regions (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 psychopathology. Some psychopathologies, such as independent major depressive disorder, bipolar disorder, anxiety disorder, or personality disorder, may be associated with alcohol consumption (Shivani, R., Goldsmith, R. J., & Anthenelli, R. M. (2002). Alcoholism and Psychiatric Disorders: Diagnostic Challenges. Alcohol Research & Health, 26(2), 90-98). The mechanism of ethanol toxicity in brain neurons has been explained, with mitochondria being the main mediator, and mitochondrial changes correlating with the severity of ethanol consumption. Therefore, improving the health of mitochondria in brain cells is being considered as a potential therapeutic target in the treatment of ethanol-related conditions (Ethanol Consumption Affects Neuronal Function: Role of the Mitochondria, Cheril Tapia-Rojas et al., December 20, 2017, DOI: 10.5772 / intechopen.71611). Also, alcohol consumption during adolescence has been shown to have a significant impact on adult mitochondrial bioenergy, indicating that this is not a temporary change until consumption ceases 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 disease lacking fine adjustment of voluntary movement. It is associated with damage to the nervous system. Mitochondrial deficiency-related ataxias include Friedreich's ataxia (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), ataxia-telangiectasia (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 00373%3Fshowall%3Dtrue#relatedArticles), etc.

[0014] 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 in mitochondrial myopath., https: / / doi.org / 10.1016 / j.mito.2020.11.005).

[0015] Brain and lung disorders associated with smoking have been demonstrated in many publications, and nicotine and tobacco smoke affect various mitochondrial activities. Mitochondria have been proposed as a possible target for the action of 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 it has been demonstrated that the action of tobacco smoke disrupts mitochondrial function and the structure of lung epithelial cells (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).

[0016] Finally, both MERRF (Myoclonic Epilepsy with Ragged Red Fibers) syndrome and NARP (Neuropathy, Ataxia, and Retinitis Pigmentosa) syndrome are associated with mitochondrial deficiency (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 p66Shc phosphorylation in fibroblasts of NAR patients).

[0017] US10792287 describes the use of pharmaceutical compounds (zolpidem supplemented with rapamycin or idebenone) for the beneficial effects on mitochondrial ATP production, inflammation suppression, and the treatment of various mitochondrial lesions such as vision loss and Leber's hereditary optic neuropathy.

[0018] In US10272056, it is also described that butyric acid-type compounds have beneficial effects on mitochondrial biosynthesis, increase in mitochondrial mass, and increase in ATP production, and are effective for several medical conditions such as neurodegenerative diseases, cardiovascular diseases, neurometabolic diseases, muscle diseases, kidney diseases, metabolic diseases, etc., and specific syndromes such as MELAS (mitochondrial encephalopathy with lactic acidosis and stroke-like episodes) and MERRF syndrome.

[0019] The applicant has surprisingly discovered that a sterol-based compound of formula (I), or a pharmaceutically acceptable salt of such a compound, is useful for the prevention, improvement and / or treatment of mitochondrial deficiency-related medical conditions caused by defects in mitochondrial dynamics and / or transport and / or activity. More precisely, the applicant has discovered that a sterol-based compound of formula (I) or a pharmaceutically acceptable salt of such a compound is useful for the prevention, improvement and / or treatment of mitochondrial deficiency-related pathologies selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychopathology, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome and NARP syndrome.

Summary of the Invention

[0020] An object of the present invention is to provide sterol-based compounds and pharmaceutical compositions containing them for use in the prevention, improvement and / or treatment of mitochondrial deficiency-related diseases selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychosis, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome, and NARP syndrome.

[0021] To achieve this, the present invention provides a compound of formula (I).

[0022]

Chemical formula

[0023] Here, R1 = OH, F, OC n H 2n+1 , OC(O)R, OC(O)OR, OC(O)NHR or OP(O)(OR)2, R = H or C n H 2n+1 , 1 ≤ n ≤ 8, and R2 = H or OH, and R3 = -NR5R6, and R5 is H or -(CH2)3NH2, and R6 is 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, and R7 = H, C(O)OCH3 or C(O)OC(CH3)3, and R4 = H or OH, at positions 20, 22, 24, 25, 26 or 27, and is arranged to form an asymmetric center of R or S configuration, Z1 and Z2 each represent the number (0 or 1) of double bonds between carbon atoms C7 and C8 and C22 and C23. T1, T2 and T3 are each independently H or CH3, and T4 is H, CH3, C2H5 arranged so as to obtain an asymmetric center of R or S configuration at the 24th position, It is used for the treatment of mitochondrial deficiency-related pathologies selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychopathology, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome and NARP syndrome.

[0024] The reference to (O) in the definition of the radical according to formula (I) means that oxygen is bonded by two bonds.

[0025] The compound of formula (I) defined by 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 is called DX243BOC or DXboc and is shown in Table 2.

[0026] The substituent or radical C(O)OC(CH3)3 is also known as the tert-butoxycarbonyl or Boc functional group.

[0027] The compounds of formula (I) belong to the steroid group. Therefore, the numbering of the carbon atoms of the compounds 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 compounds belonging to the steroid group according to IUPAC is shown below.

[0028]

Chemical formula

[0029] The method for preparing the compounds of formula (I) has already been described 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.

[0030] Furthermore, the compounds may have one or more of the following characteristics, alone or in combination. According to one embodiment, the compound of formula (I) is such that R1 = OH, F, OC n H 2n+1 , OC(O)R, OC(O)OR, OC(O)NHR or OP(O)(OR)2 (R = H or C n H 2n+1 , 1 ≤ n ≤ 8), and R2 = OH, R3 is -NR5R6, R5 is H, R6 is -(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, and R7 is H, C(O)OCH3 or C(O)OC(CH3)3, Z1 is 0 or 1, Z2 is 0, R4 is H.

[0031] According to one embodiment, the compound of formula (I) is more precisely Z1 = 0, Z2 = 0, R1 = R2 = OH, R4 = H, R5 = H, T1 = T2 = T3 = T4 = H, and the other radicals R3, R6 and R7 are as defined above.

[0032] According to one embodiment, the compound of formula (I) is more precisely Z1 = 0, Z2 = 0, R1 = R2 = OH, R4 = H, R5 = H, T1 = T2 = T3 = T4 = H, R6 = -(CH2)4NH(CH2)3NHR7 and R7 = C(O)CH3. This compound, named DX249, is 5α-hydroxy-6β-[3-(4-aminobutylacetamido)propylamino]cholestan-3β-ol.

[0033] According to one embodiment, the compound of formula (I) is more precisely Z1 = 0, Z2 = 0, R1 = R2 = OH, R4 = H, R5 = H, T1 = T2 = T3 = T4 = H, R6 = -(CH2)2-imidazol-4-yl. This compound, named DX101, is 5α-hydroxy-6β-[2-(1H-imidazol-4-yl)ethylamino]cholestan-3β-ol.

[0034] According to one embodiment, the compound of formula (I) is more precisely Z1 = 0, Z2 = 0, R1 = R2 = OH, R4 = H, R5 = H, T1 = T2 = T3 = T4 = H, R6 = -(CH2)3NR7(CH2)4NHR7, -(CH2)4NR7(CH2)3NHR7, -(CH2)3NR7(CH2)4NH(CH2)3NHR7, or -(CH2)4NHR7, and R7 = H. These compounds, named DX243, DX245, DX301, and DX401 respectively, are as follows. -5α-Hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholestan-3β-ol (DX243) -5α-Hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholestan-3β-ol (DX245) -5α-Hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholestan-3β-ol (DX301) -5α-Hydroxy-6β-(4-aminobutylamino)cholestan-3β-ol (DX401)

[0035] According to one embodiment, the compound of formula (I) is more precisely Z1 = 0, Z2 = 0, R1 = R2 = OH, R4 = H, R5 = H, T1 = T2 = T3 = T4 = H, R6 = -(CH2)3NR7(CH2)4NHR7, R7 = C(O)OC(CH3)3. This compound, named DX243BOC, is 5α-hydroxy-6β-[3-(4-tert-butyloxycarbonylaminobutyl-tert-butyloxycarbonylamino)propylamino]cholestan-3β-ol.

[0036] According to one embodiment, the compound of formula (I) is more precisely Z1 = 1, Z2 = 0, R1 = R2 = OH, R4 = H, R5 = H, T1 = T2 = T3 = T4 = H, and the other radicals R3, R6, and R7 are defined as described above.

[0037] According to one embodiment, the compound of formula (I) is, more precisely, Z1 = 1, Z2 = 0, R1 = R2 = OH, R4 = H, R5 = H, T1 = T2 = T3 = T4 = H, R6 = -(CH2)3NH(CH2)4NHR7, -(CH2)4NH(CH2)3NHR7, or -(CH2)3NH(CH2)4NH(CH2)3NHR7, and R7 = H. These compounds, named DX242, DX244, and DX302 respectively, are as follows. -5α-Hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholest-7-en-3β-ol (DX242) -5α-Hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholest-7-en-3β-ol (DX244) -5α-Hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholest-7-en-3β-ol (DX302)

[0038] According to one embodiment, the mitochondrial deficiency-related pathology is due to a deficiency in mitochondrial dynamics and / or transport and / or activity. Table 1 shows the specificity of the deficiency according to the pathology.

[0039] [Table 1]

[0040] In the case of the compound of formula (I) according to the present invention, the mitochondrial deficiency-related pathology is due to a deficiency in mitochondrial dynamics and / or mitochondrial transport and / or mitochondrial activity.

[0041] In one embodiment, the pathologies associated with deficiencies in mitochondrial dynamics and mitochondrial transport and activity are neuropathy, spinal cord injury, autism, ataxia, and alcohol-related psychopathology.

[0042] In another embodiment, the pathologies associated with mitochondrial dynamics and lack of mitochondrial activity are migraine, smoking-related brain and lung disorders, and MERRF syndrome.

[0043] In another embodiment, the pathology associated with lack of mitochondrial transport and activity is multiple sclerosis.

[0044] In yet another embodiment, the medical condition associated with lack of mitochondrial activity is NARP syndrome.

[0045] A second subject of the present invention is 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 deficiency-related medical conditions 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.

[0046] The compositions used according to the present invention can be administered in various forms suitable for the medical condition to be treated. Thus, the various administration forms include oral, topical, systemic, intravenous, subcutaneous, intraperitoneal, intramuscular, transdermal or transmucosal administration.

[0047] The compositions of the present invention can be in any form commonly used depending on whether they are for ingestion, injection, or application to the skin or mucosa.

[0048] The compositions of the present invention can include components commonly used in this type of formulation, such as binders, flavorings, preservatives, colorants, etc., and in the case of food supplements or pharmaceuticals, can be in the form of tablets, granules, or gel capsules. The formulations of the present invention can be in the form of foods such as beverages, or in the form of suspensions or syrups.

[0049] According to the first variant, various formulations are suitable for topical administration and include creams, water-in-oil and oil-in-water emulsions, milks, ointments, lotions, oils, balms, aqueous or aqueous-alcoholic or glycolic acid solutions, serums, powders, patches, sprays, or other products for external application such as medical devices or aerosol products containing a pressurized propellant.

[0050] According to the second variant, various compositions are suitable for injection and the composition may be in the form of an aqueous or oily lotion or in the form of a serum.

[0051] According to the third variant, various compositions are suitable for ingestion and the composition may be in the form of capsules, syrups, granules or tablets.

[0052] According to a preferred embodiment, the compositions of the present invention are more specifically intended for topical administration. Thus, these compositions must include a dermatologically acceptable medium, i.e., a medium that is compatible with the skin and mucous membranes and covers all dermatological forms. These compositions may in particular be in the form of creams, water-in-oil or oil-in-water emulsions or multiple emulsions, serums, solutions, suspensions, gels, milks, lotions, sticks or powders and may be suitable for application to the skin and mucous membranes. These compositions include excipients necessary for their formulation, such as solvents, emollients, thickeners, diluents, surfactants, antioxidants, bioactive agents, dyes, preservatives, fragrances, etc.

[0053] The compositions of the present invention also include any additives commonly used in the envisaged fields of application, as well as adjuvants necessary for their formulation, such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens, self-tanning agents, pigments, fillers, preservatives, fragrances, odor absorbers, dermatological or pharmaceutical active agents, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0054] In any case, the person skilled in the art takes care to ensure that these adjuvants and their proportions are selected so that the desired advantageous properties of the compositions of the present invention are not adversely affected.

[0055] According to one embodiment, the composition used according to the present invention is in the form of an aqueous solution, and the concentration of the compound of formula (I) is 1 pmol·L -1 ~1 mmol·L -1 , preferably 10 pmol·L -1 ~0.1 mmol·L -1 , more preferably 0.1 nmol·L -1 ~1 μmol·L -1 .

[0056] With reference to the accompanying drawings, from the following description of some specific embodiments of the present invention shown purely by way of non-limiting example, the present invention will be better understood and other objects, details, features and advantages of the present invention will become more clearly apparent.

Brief Description of the Drawings

[0057]

Figure 1

Figure 2A

Figure 2B

Figure 2C

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

DETAILED DESCRIPTION OF THE INVENTION

[0058] In this description, unless otherwise specified, when a range is indicated, it is understood that the range includes its upper and lower limits.

[0059] In the present invention, the following definitions apply:

[0060] "Prevention of a medical condition" means avoiding the occurrence of a disease or injury, or maintaining and improving health. It is also called preventive treatment.

[0061] "Improvement of a medical condition" means reducing the symptoms of the said medical condition. It is also called palliative treatment.

[0062] "Treatment of a medical condition" means curing a disease. It is also called curative treatment.

[0063] "Mitochondrial deficiency" means that the activity of mitochondria and / or the dynamics of mitochondria and / or the axonal transport of mitochondria decrease, energy production is lost, and harmful substances accumulate in the body.

[0064] "Activity of mitochondria" means the ability of mitochondria to regulate cell metabolism and produce energy (ATP) via the respiratory chain.

[0065] "Mitochondrial dynamics" means the ability of mitochondria to fuse or divide to maintain their shape and size and / or increase the amount (mass) of mitochondria in the cell.

[0066] "Mitochondrial axonal transport" means the ability of mitochondria to move to meet the necessary energy demands at precise locations within the cell.

[0067] "Mitochondrial fission" means that mitochondria divide into two different mitochondria.

[0068] To prove that the compound of formula (I) affects the mitochondrial function of cells, various experiments were conducted to demonstrate the beneficial effect of said compound on mitochondrial function in general, through its effects on the number, dynamics, stress, transport or activity of mitochondria. Below, several experimental protocols for demonstrating the beneficial effects of the compounds of formula (I) shown in Table 2 below, in particular, under various conditions and experimental models, will be described.

[0069]

Table 2

[0070] The concentration or molar concentration of the compound is expressed in moles per liter, and its symbol is mol.L -1 or M.

[0071] [Example 1: Test of relative metabolic activity on cortical neurons]

[0072] A protocol was developed to obtain a primary culture of cortical neurons from cells collected from the brains of wild-type mouse embryos. Step 1 was to collect the embryonic cortex and place the cortical neurons under appropriate culture conditions. Step 2 was to culture the collected cells in NeurobasalTM medium (Ref. 21103049 ThermoFisher Scientific), to which L-glutamine and B27 supplement 50X (Ref. 17504044 ThermoFisher Scientific) were added. Next, the neurons were separated and purified from the primary culture. Under the above culture conditions, a neuron culture purified from the separation of the embryonic cortex was obtained.

[0073] Tests based on the metabolic activity of neurons were conducted to evaluate the effect of the compounds of formula (I) on this metabolic activity. This test is based on the use of the tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). Tetrazolium is reduced by mitochondrial succinate dehydrogenase in active living cells to formazan, a purple precipitate. The amount of precipitate formed is proportional to the metabolic activity present in the culture. Thus, it is possible to determine the relative amount of metabolic activity and, by extension, mitochondrial activity by spectrophotometric analysis of the optical density at 550 nm. The results obtained are shown in Figure 1. This test on cortical neuron cultures was conducted using different compounds (DX101, DX243, DX244, DX245, DX249, DX301, DX302, and DX401) at the same concentration (100 nM) with reference to Table 2.

[0074] Referring to Figure 1, this test can highlight the positive effect of the test compounds of formula (I) on mitochondrial activity as compared to untreated neurons.

[0075] [Example 2: Test of relative metabolic activity on Neuro-2a neurons]

[0076] The same test as described in Example 1 was conducted on Neuro-2a cells, a mouse neuroblastoma cell line. Under physiological conditions, at approximately 50% confluence after culturing, Neuro-2a cells were contacted with two concentrations of compounds DX243 and DX245 for 24 hours. The MTT assay performed after 24 hours of treatment demonstrated that compound DX243, and particularly compound DX245, increased the metabolic activity of Neuro-2a cells, as shown in Figure 2A.

[0077] [Example 3: Test of metabolic activity after ethanol (EtOH) intoxication]

[0078] The same test described in Example 1 was performed on Neuro-2a cells, a mouse neuroblastoma cell line. When these cells were intoxicated with a high concentration (500 mM) of ethanol (EtOH) for 24 hours, it was shown that the metabolic activity of these cells decreased (X = P value < 0.0001 compared to the control). This metabolic activity was partially restored by treatment with 1 nM of compound DX243 for 24 hours.

[0079] Advantageously, as shown in Figure 2B, after intoxication with EtOH (200 mM and 500 mM), treatment with 100 nM of compound DX243 or compound DX245 for 24 hours resulted in a significantly higher metabolic activity compared to untreated cells.

[0080] Figure 2C shows the effect of compound DX243 on mitochondria in Neuro-2a cells after ethanol intoxication. This figure shows the results of examining the increase in mitochondrial surface area in the presence of compound DX243 24 hours after ethanol intoxication (500 mM), with the help of MitoTracker.

[0081] [- Example 4: Tests on mitochondrial dynamics in SHSY5Y and C2C12 cell lines]

[0082] Human neuroblastoma cell line (SHSY5y) and myoblast cell line (C2C12) were cultured in DMEM (trademark) medium (Ref. 21068028 ThermoFisher Scientific). After culturing, these two cell lines were treated with different concentrations of compound DX243 for 24 hours. A series of images were taken and labeled with Mitotracker (trademark) (Ref. M7512, ThermoFisher Scientific). This labeling highlights and visualizes the mitochondria present in these cells, enabling quantification of their amount. Quantification of the mitochondrial mass revealed by Mitotracker labeling emphasized the effect of compound DX243 on mitochondrial dynamics (an increase in the amount (mass) of mitochondria) (see Figure 3).

[0083] [Example 5: Test for evaluating inhibition of mitochondrial fission in snail tissue slices]

[0084] A snail tissue slice model was developed. To obtain cultured snails, which are auditory organs, the snails of 3-day-old mice were microdissected. Next, these snails were placed in DMEM medium supplemented with glucose (0.5%), N1 (0.5%), insulin (0.25%), and penicillin (0.1%) for 24 hours. Next, 100 nM of compound DX243 was added to this medium for 24 hours.

[0085] Next, the snails were collected, and RNA was extracted from them so as to enable analysis of the modulation of several genes after treatment with compound DX243. As a result, as shown in Figure 4, it was observed that when the snail tissue slices were treated with compound DX243 (100 nM), the expression of the mitochondrial fission gene drp1 was inhibited. This test demonstrates that compound DX243 plays a beneficial role in mitochondria, particularly by inhibiting mitochondrial fission.

[0086] [Example 6: Test for evaluating mitochondrial dynamics and mitochondrial stress in dopaminergic neurons]

[0087] The mitochondrial effect of compound DX243 was tested in a model of dopaminergic neurons treated with MPP+ (1-methyl-4-phenylpyridinium), a mitochondrial toxin that increases mitochondrial stress and reduces the number of mitochondria. MPP+ is known to have a harmful effect on mitochondria (increase in mitochondrial stress measured by the amount of cytochrome C, decrease in the number of mitochondria measured by the Mitotracker assay). Dopaminergic neurons were obtained from the microdissection of the midbrain of fetal rats cultured in Neurobasal™ medium (Ref. 21103049 ThermoFisher Scientific) supplemented with B27 supplement 50X (Ref. 17504044 ThermoFisher Scientific), penicillin / streptomycin, L-glutamine, and BDNF (brain-derived neurotrophic factor) and GDNF (glial cell-derived neurotrophic factor). These neurons were cultured for 5 days and then treated with MPP + for 48 hours. Also, these neurons were exposed to various concentrations of compound DX243 during the 48 hours of MPP + treatment and 48 hours after the removal of MPP + .

[0088] The results, as shown in Figure 5, indicate that compound DX243 can increase and recover the mitochondrial mass of these dopaminergic neurons that have lost a part of their mitochondrial mass after MPP+ treatment.

[0089] Furthermore, as shown in Figure 6, 10 nM of compound DX243 also brings about a reduction in mitochondrial stress caused by MPP + treatment.

[0090] [Example 7: Test for evaluating mitochondrial dynamics and mitochondrial stress in hippocampal neurons]

[0091] The mitochondrial effect of compound DX243 was tested in a hippocampal neuron model treated with peptide Aβ1-42, which induces mitochondrial stress (represented by a decrease in mitochondrial surface area). As shown in Figure 7, compound DX243 (1 nM) was shown to be able to restore this decrease in mitochondrial surface area.

[0092] The use of the verbs "comprise", "comprising" or "comprised of" and their conjugations does not exclude the presence of elements or steps other than those recited in the claims.

[0093] In the claims, reference signs in parentheses shall not be construed as limiting the claim.

Claims

1. A compound of formula (I). 【Chemical 1】 Here, R １ = 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 , 1 ≤ n ≤ 8, R ２ = H or OH, and R ３ = -NR ５ R ６ wherein R ５ is H or -(CH ２ ) ３ NH ２ and, R ６ is selected from the group consisting of -(CH ２ ), ３ NR ７ (CH ２ ), ４ NHR ７ -(CH ２ ), ３ NHR ７ -(CH ２ ), ４ NHR ７ -(CH ２ ), ４ NR ７ (CH ２ ), ３ NHR ７ -(CH ２ ), ３ NR ７ (CH ２ ), ４ NR ７ (CH ２ ), ３ NHR ７ -(CH ２ ), ２ -imidazol-4-yl,-(CH ２ ), ２ -indol-3-yl, R ７ =H, C(O)OCH ３ or C(O)OC(CH ３ ), ３ and R ４ is H or OH at the 20, 22, 24, 25, 26 or 27 position and is arranged to form an asymmetric center of the R or S configuration, Z １ and Z ２ represent the number of double bonds between carbon atoms C7 and C8 and C22 and C23, respectively (0 or 1), T １ T ２ and T ３ are independently of one another H or CH ３ and T ４ is H, CH ３ C ２ H ５ and is arranged to form an asymmetric center with an R or S configuration at position 24 It is used for the treatment of mitochondrial deficiency-related pathologies selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychopathology, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome, and NARP syndrome.

2. The compound of formula (I) is R １ = OH, F, OC ｎ H ２ｎ＋１ , OC(O)R, OC(O)OR, OC(O)NHR or OP(O)(OR) ２ , R = H or C ｎ H ２ｎ＋１ , 1 ≤ n ≤ 8, and R ２ = OH, and R ３ = -NR ５ R ６ wherein R ５ = H, and R ６ = -(CH ２ ) ３ NR ７ (CH ２ ) ４ NH ７ 、-(CH ２ ) ３ NH ７ 、-(CH ２ ) ４ NH ７ 、-(CH ２ ) ４ NR ７ (CH ２ ) ３ NH ７ 、-(CH ２ ) ３ NR ７ (CH ２ ) ４ NR ７ (CH ２ ) ３ NH ７ 、-(CH ２ ) ２ -imidazol-4-yl, -(CH ２ ) ２ -indol-3-yl, R ７ = H, C(O)OCH ３ or C(O)OC(CH ３ ) ３ and, Z １ is 0 or 1, and Z ２ = 0, and R ４ A compound for use according to claim 1, as defined by R = H.

3. The compound of formula (I) is Z １ = 0, Z ２ = 0, R １ = R ２ = OH, R ４ = H, R ５ = H, T １ = T ２ = T ３ = T ４ A compound for use according to claim 1 or 2, defined as = H

4. The compound of formula (I) is R ６ = -(CH ２ ) ４ NH(CH ２ ) ３ NHR ７ , R ７ = C(O)CH ３ as defined, and is the compound 5α-hydroxy-6β-[3-(4-aminobutylacetamido)propylamino]cholestan-3β-ol, the compound for use according to claim 3.

5. The compound of formula (I) is R ６ = -(CH ２ ) ２ NHR ７ 、R ７ is defined as imidazol-4-yl, and the compound is 5α-hydroxy-6β-[2-(1H-imidazol-4-yl)ethylamino]cholestan-3β-ol, a compound for use according to claim 3.

6. The compound of formula (I) is R ６ = -(CH ２ ) ３ NR ７ (CH ２ ) ４ NHR ７ -(CH ２ ) ４ NR ７ (CH ２ ) ３ NHR ７ -(CH ２ ) ３ NR ７ (CH ２ ) ４ NR ７ (CH ２ ) ３ NHR ７ or -(CH ２ ) ４ NHR ７ and R ７ is defined by R = H, and is 5α-hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholestan-3β-ol, 5α-hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholestan-3β-ol, 5α-hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholestan-3β-ol, 5α-hydroxy-6β-(4-aminobutylamino)cholestan-3β-ol, respectively, a compound for use according to claim 3.

7. The compound of formula (I) is R ６ = -(CH ２ ) ３ NR ７ (CH ２ ) ４ NHR ７ and R ７ = C(O)OC(CH ３ ) ３ as defined, and is 5α-hydroxy-6β-[3-(4-tert-butyloxycarbonylaminobutyl-tert-butyloxycarbonylamino)propylamino]cholestan-3β-ol, a compound for use according to claim 3.

8. The compound of formula (I) is Z １ = 1 and Z ２ = 0, R １ = R ２ = OH, R ４ = H, R ５ = H, T １ = T ２ = T ３ = T ４ A compound for use according to claim 2, defined as = H

9. The compound of formula (I) is R ６ = -(CH ２ ) ３ NR ７ (CH ２ ) ４ NHR ７ -(CH ２ ) ４ NR ７ (CH ２ ) ３ NHR ７ or -(CH ２ ) ３ NR ７ (CH ２ ) ４ NR ７ (CH ２ ) ３ NHR ７ and R ７ = H, and are respectively 5α-hydroxy-6β-[3-(4-aminobutylamino)propylamino]cholest-7-en-3β-ol, 5α-hydroxy-6β-[4-(3-aminobutylamino)propylamino]cholest-7-en-3β-ol, 5α-hydroxy-6β-{3-[4-(3-aminopropylamino)butylamino]propylamino}cholest-7-en-3β-ol, the compound for use according to claim 8.

10. A compound for use according to any one of claims 1 to 9, wherein the mitochondrial deficiency-related pathology is caused by a deficiency 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 mitochondrial deficiency-related pathologies selected from the group consisting of autism, spinal cord injury, multiple sclerosis, epilepsy, migraine, alcohol-related psychopathology, ataxia, neuropathy, smoking-related brain and lung disorders, MERRF syndrome, and NARP syndrome.

12. In the form of an aqueous solution, wherein the concentration of the compound of formula (I) is 1 pmol·L －１ to 1 mmol·L －１ , preferably 10 pmol·L －１ to 0.1 mmol·L －１ , more preferably 0.1 nmol·L －１ to 1 μmol·L －１ The composition according to claim 11, characterized in that it is so.

Images