Hydroxybenzoic acid derivatives, methods and uses thereof

Abstract

Hydroxybenzoic acid-based derivatives and analogs are provided for treating, for example, mitochondrial dysfunction or deficiency in the field of human and animal diseases. For example, the compound is represented by the following formula: JPEG2023051961000017.jpg4981 (R 1 ~R 5 is H, halogen, hydroxyl, methyl, etc.; R 7 represents an alkyl chain, an alkenyl chain, etc.; Z - is an acceptable anion.

Description

[Technical field]

[0001] The present disclosure provides novel mitochondrial-tropic antioxidants based on hydroxybenzoic acids and analogues. Further, the present disclosure relates to the design and synthesis of compounds for the treatment of, e.g., human and animal diseases. in the field of, for example, treating mitochondrial dysfunction or mitochondrial deficiency Methods and uses of hydroxybenzoic acid-based derivatives and analogs for the treatment of cancer, e.g. For example, it relates to cosmetics for preventing or delaying skin aging. [Background technology]

[0002] Mitochondria regulate energy metabolism, cytosolic calcium concentration, ROS production, and Plays a pivotal role in regulating cell death pathways. Excessive ROS production inhibits the innate defense If not prevented by mechanisms, it can cause oxidative damage to cellular components such as lipids, proteins, and nucleic acids. It can cause injury, which can lead to subsequent cell death by necrosis or apoptosis. Flow.

[0003] Mitochondrial alterations resulting from enhanced oxidative stress are associated with a number of risk factors including cancer, stroke, heart failure, obesity and It plays an important role in oxidative stress-related diseases such as osteoporosis and neurodegenerative diseases. 1 Organ Targeting mitochondria with Ras-specific drugs is believed to be an effective therapeutic strategy. More specifically, we aim to improve cellular ROS balance through selective delivery of antioxidants to mitochondria. Is regulating oxidative stress an effective method for preventing and / or treating oxidative stress-related diseases? has been described as a promising treatment strategy. 2 .

[0004] ROS production is tightly regulated by the endogenous antioxidant network, but its overproduction can result in oxidative damage and dysfunction of mitochondria. Oxidative mitochondrial dysfunction impairs multiple metabolic and signaling pathways and may cause cell death via apoptosis or necrosis.

[0005] Oxidative stress and mitochondrial dysfunction are associated with aging and several oxidative stress-related pathologies, such as diabetes, non-alcoholic fatty liver disease, cardiovascular disease, acute pancreatitis, and neurodegenerative diseases including Alzheimer's disease or Parkinson's disease, as well as amyotrophic lateral sclerosis. Therefore, prevention of oxidative damage to mitochondria is currently

[0006] an accepted pharmacological strategy for retarding disease progression. In pathological events, the pool of endogenous antioxidant defenses may not be sufficient to cope with increased oxidant production, and considering that exogenous antioxidants not only supplement the inadequacy of the endogenous defense system but also improve the overall antioxidant response, it has been suggested that administration of exogenous antioxidants may be beneficial in reducing cellular injury. Exogenous antioxidants could theoretically interrupt the complex network of oxidative damage pathways at different levels

[0007] and result in a therapeutic effect. As a result, antioxidants obtained exogenously from the diet may have important functions in redox cell Improvement of mitochondrial function by ROS / antioxidants is an effective and promising therapeutic strategy. Maintaining the ratio and redox balance is important for cell signaling, so targeting antioxidants to dysfunctional mitochondria is pharmacologically interesting. Since maintaining the ratio and redox balance is important for cell signaling, targeting antioxidants to dysfunctional mitochondria is pharmacologically interesting.

[0008] A number of mitochondrially targeted antioxidants, especially those using triphenylphosphonium (TPP) as a carrier, have been developed. This type of lipophilic cation can cross the mitochondrial membrane using the inner membrane potential gradient and accumulate within the mitochondrial matrix. A number of mitochondrially targeted antioxidants, especially those using triphenylphosphonium (TPP) as a carrier, have been developed. This type of lipophilic cation can cross the mitochondrial membrane using the inner membrane potential gradient and accumulate within the mitochondrial matrix. A number of mitochondrially targeted antioxidants, especially those using triphenylphosphonium (TPP) as a carrier, have been developed. This type of lipophilic cation can cross the mitochondrial membrane using the inner membrane potential gradient and accumulate within the mitochondrial matrix. A number of mitochondrially targeted antioxidants, especially those using triphenylphosphonium (TPP) as a carrier, have been developed. This type of lipophilic cation can cross the mitochondrial membrane using the inner membrane potential gradient and accumulate within the mitochondrial matrix. 3~5 .

[0009] One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. One of the most studied mitochondrially targeted antioxidants is mitoquinone (MitoQ, MitoQ10, [10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl]triphenylphosphonium methanesulfonate). MitoQ is composed of an endogenous antioxidant moiety (coenzyme Q) covalently linked to a 10-carbon alkyl chain (dTPP) spacer and a triphenylphosphonium (TPP) cation. MitoQ is in clinical trials for various pathological events, namely hepatitis C. However, disappointing results have been obtained in clinical trials using MitoQ as a therapeutic agent for neurodegenerative diseases. 4,5 .

[0010] Another related mitochondrially targeted antioxidant is SKQ1 [10-(4,5-dimethyl-3,6-dioxocyclohex-1,4-dien-1-yl)decyl)triphenylphosphonium bromide)], which is involved in the electron transport chain of plastoquinone, chloroplasts. Another related mitochondrially targeted antioxidant is SKQ1 [10-(4,5-dimethyl-3,6-dioxocyclohex-1,4-dien-1-yl)decyl)triphenylphosphonium bromide)], which is involved in the electron transport chain of plastoquinone, chloroplasts. Another related mitochondrially targeted antioxidant is SKQ1 [10-(4,5-dimethyl-3,6-dioxocyclohex-1,4-dien-1-yl)decyl)triphenylphosphonium bromide)], which is involved in the electron transport chain of plastoquinone, chloroplasts. It is based on quinone. SkQ1 has been shown to reduce oxidative stress in mitochondria and provide a significant protective effect against the dry state. It has been shown to provide a significant protective effect against the dry state.

[0011] Nevertheless, there is still a need for more effective and safe mitochondrial modulators for use in therapy, as well as in other applications such as supplements or nutraceuticals, and in the cosmetic field. Nevertheless, there is still a need for more effective and safe mitochondrial modulators for use in therapy, as well as in other applications such as supplements or nutraceuticals, and in the cosmetic field. There is still a need for more effective and safe mitochondrial modulators for use in therapy, as well as in other applications such as supplements or nutraceuticals, and in the cosmetic field.

[0012] Polyphenols are secondary metabolites of plants that are widely found in fruits, vegetables, grains, and beverages that make up the human diet and are generally involved in the defense against oxidative stress. Their daily dietary intake in a conventional Western diet was estimated to be about 1 g. Epidemiological studies and related meta-analyses strongly suggest an association between the intake of a polyphenol-rich diet and the prevention of oxidative stress-related diseases such as cancer, diabetes, cardiovascular disease, and neurodegenerative diseases. Polyphenols are secondary metabolites of plants that are widely found in fruits, vegetables, grains, and beverages that make up the human diet and are generally involved in the defense against oxidative stress. Their daily dietary intake in a conventional Western diet was estimated to be about 1 g. Epidemiological studies and related meta-analyses strongly suggest an association between the intake of a polyphenol-rich diet and the prevention of oxidative stress-related diseases such as cancer, diabetes, cardiovascular disease, and neurodegenerative diseases. Polyphenols are secondary metabolites of plants that are widely found in fruits, vegetables, grains, and beverages that make up the human diet and are generally involved in the defense against oxidative stress. Their daily dietary intake in a conventional Western diet was estimated to be about 1 g. Epidemiological studies and related meta-analyses strongly suggest an association between the intake of a polyphenol-rich diet and the prevention of oxidative stress-related diseases such as cancer, diabetes, cardiovascular disease, and neurodegenerative diseases. Polyphenols are secondary metabolites of plants that are widely found in fruits, vegetables, grains, and beverages that make up the human diet and are generally involved in the defense against oxidative stress. Their daily dietary intake in a conventional Western diet was estimated to be about 1 g. Epidemiological studies and related meta-analyses strongly suggest an association between the intake of a polyphenol-rich diet and the prevention of oxidative stress-related diseases such as cancer, diabetes, cardiovascular disease, and neurodegenerative diseases. Polyphenols are secondary metabolites of plants that are widely found in fruits, vegetables, grains, and beverages that make up the human diet and are generally involved in the defense against oxidative stress. Their daily dietary intake in a conventional Western diet was estimated to be about 1 g. Epidemiological studies and related meta-analyses strongly suggest an association between the intake of a polyphenol-rich diet and the prevention of oxidative stress-related diseases such as cancer, diabetes, cardiovascular disease, and neurodegenerative diseases.

[0013] Hydroxybenzoic acid (HBA), a subclass of phenolic acids, contains seven carbon atoms (C6-C1) bonded to at least one hydroxyl group. Some HBA derivatives are currently used as food antioxidant additives to prevent or minimize the oxidation of nutrients and maintain or improve the nutritional value of foods. Hydroxybenzoic acid and its derivatives are also used as excipients in the cosmetic and pharmaceutical industries due to their antioxidant properties. However, some drawbacks have been pointed out mainly related to their effectiveness. Hydroxybenzoic acid (HBA), a subclass of phenolic acids, contains seven carbon atoms (C6-C1) bonded to at least one hydroxyl group. Some HBA derivatives are currently used as food antioxidant additives to prevent or minimize the oxidation of nutrients and maintain or improve the nutritional value of foods. Hydroxybenzoic acid and its derivatives are also used as excipients in the cosmetic and pharmaceutical industries due to their antioxidant properties. However, some drawbacks have been pointed out mainly related to their effectiveness. Hydroxybenzoic acid (HBA), a subclass of phenolic acids, contains seven carbon atoms (C6-C1) bonded to at least one hydroxyl group. Some HBA derivatives are currently used as food antioxidant additives to prevent or minimize the oxidation of nutrients and maintain or improve the nutritional value of foods. Hydroxybenzoic acid and its derivatives are also used as excipients in the cosmetic and pharmaceutical industries due to their antioxidant properties. However, some drawbacks have been pointed out mainly related to their effectiveness. Hydroxybenzoic acid (HBA), a subclass of phenolic acids, contains seven carbon atoms (C6-C1) bonded to at least one hydroxyl group. Some HBA derivatives are currently used as food antioxidant additives to prevent or minimize the oxidation of nutrients and maintain or improve the nutritional value of foods. Hydroxybenzoic acid and its derivatives are also used as excipients in the cosmetic and pharmaceutical industries due to their antioxidant properties. However, some drawbacks have been pointed out mainly related to their effectiveness. Hydroxybenzoic acid (HBA), a subclass of phenolic acids, contains seven carbon atoms (C6-C1) bonded to at least one hydroxyl group. Some HBA derivatives are currently used as food antioxidant additives to prevent or minimize the oxidation of nutrients and maintain or improve the nutritional value of foods. Hydroxybenzoic acid and its derivatives are also used as excipients in the cosmetic and pharmaceutical industries due to their antioxidant properties. However, some drawbacks have been pointed out mainly related to their effectiveness. Hydroxybenzoic acid (HBA), a subclass of phenolic acids, contains seven carbon atoms (C6-C1) bonded to at least one hydroxyl group. Some HBA derivatives are currently used as food antioxidant additives to prevent or minimize the oxidation of nutrients and maintain or improve the nutritional value of foods. Hydroxybenzoic acid and its derivatives are also used as excipients in the cosmetic and pharmaceutical industries due to their antioxidant properties. However, some drawbacks have been pointed out mainly related to their effectiveness. However, some drawbacks have been pointed out mainly related to their effectiveness.

[0014] The antioxidant activity of HBA is related to their chelating and free radical scavenging properties, and That is, it is related by inhibiting the lipid peroxidation process. HBA derivatives play a role in the inhibition of several oxidation-promoting enzymes involved in the production of reactive oxygen species (ROS), which is now also recognized. According to scientific evidence, the antioxidant effect of HBA is related to the number and position of hydroxyl groups on the aromatic ring. These different mechanisms of action can result in the inhibition or reduction of ROS formation, the interference with the propagation of free radical chain reactions, or the delay of their initiation or

[0015] reaction rate. The usefulness of HBA and their derivatives, either alone or as adjuvants, in therapy is mainly limited due to the limitations of bioavailability and efficacy. Despite their putative health-promoting properties, the bioavailability of orally administered polyphenols is insufficient to allow sufficient concentrations for systemic therapy, mainly due to their physicochemical properties (e.g., lipophilicity) and problems related to extensive and rapid metabolism. To enhance the lipophilicity and stability of HBA, enable better bioavailability, and improve their delivery to intracellular

[0016] targets such as mitochondria, various strategies have been developed so far. These facts are disclosed to illustrate the technical problems addressed by the present disclosure.

[0017] General description Mitochondria are attractive targets for many molecules, which can minimize organelle damage in different pathological situations.

[0018] Increasing evidence shows that oxidation plays a decisive role in various pathologies and aging suggests that mitochondrial dysfunction amplifies stress events. Mitochondria iron-sulfur centers, membrane polyunsaturated fatty acids, proteins, and mitochondrial DNA are susceptible to oxidative damage and often cause organelle and cell destruction.

SUMMARY OF THE INVENTION

PROBLEMS TO BE SOLVED BY THE INVENTION

[0019] The present disclosure reports the design and synthesis of novel mitochondria-targeted antioxidants based on dietary hydroxybenzoic acids and analogs (AntiOxBEN).

[0020] As part of the present disclosure related to the development of effective antioxidants based on natural models, herein reports the development of novel mitochondria-directed antioxidants based on natural dietary HBA that possess strong antioxidant and iron chelation properties while maintaining a low cytotoxicity profile.

MEANS FOR SOLVING THE PROBLEMS

[0021] The present disclosure relates to a compound of formula I, or a salt, solvate, hydrate, tautomer, stereoisomer; preferably a pharmaceutically acceptable salt, solvate, hydrate, tautomer, stereoisomer for use in medicine:

CHEM.

[0022] The present disclosure relates to a compound of formula I, or a salt, solvate, hydrate, tautomer, stereoisomer; preferably a pharmaceutically acceptable salt, solvate, hydrate, tautomer, stereoisomer:

Chemical formula

[0023] According to the definition of the International Union of Pure and Applied Chemistry (IUPAC), an alkyl group is any carbon atom. A monovalent radical -C derived from an alkane by removing a hydrogen atom from the n H 2n+ 1 It is defined as the unit of alkane olefins, which are formed by removing hydrogen atoms from the terminal carbon atoms of unbranched alkanes. The radical derived from this is the straight chain alkyl (n-alkyl) radical H(CH 2 ) n Form a subclass of RCH 2 , R 2 CH (R is not H), and R 3 C (R is not H) is These are primary, secondary and tertiary alkyl groups, respectively. Aryl groups are groups that detach a hydrogen atom from a ring carbon atom. They are derived from arenes (monocyclic and polycyclic aromatic hydrocarbons) by removal of

[0024] "Alkyl" includes "lower alkyl", and refers to any carbon fragment having 30 or fewer carbon atoms. Examples of alkyl groups include octyl, nonyl, norbornyl, and the like. , undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, eicosyl, 3, 7-Diethyl-2,2-dimethyl-4-propylnonyl, 2-(cyclododecyl)ethyl , adamantyl, and the like.

[0025] "Lower alkyl" means an alkyl group of 1 to 7 carbon atoms. Examples include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and te tert-butyl, pentyl, hexyl, heptyl, cyclopropyl, cyclobutyl, cyclo pentyl, cyclohexyl, cycloheptyl, 2-methylcyclopropyl, cyclopropi lumethyl and the like are included.

[0026] In one embodiment, the compound of formula I is as follows:

Chemical formula

[0027] In one embodiment, the compound of formula I is as follows:

Chemical formula

[0028] In one embodiment, R 7 is a secondary amide of R 8 -(C=O)NH-R 9 amide; R 8 and R 9 are independently selected from each other; R 8 and R 9 are an alkyl chain, an alkenyl chain, an alkynyl chain or a substituted aryl .

[0029] In one embodiment, the substituted aryl is alkane-aryl substitution, alkene-aryl substitution , or alkyne-aryl substitution.

[0030] In one embodiment, the alkane-aryl substitution, alkene-aryl substitution, or alkyne -aryl substitution is as follows. C 1 ~C 6 -alkyl, C 3 ~C 8 -cycloalkyl, C 6 ~C 10 -aryl, C 6 ~C 10 -aryl-C 1 ~C 8 -alkyl, C 1 ~C 6 -alkoxy, C 6 ~C 10 - aryloxy, C 6 ~C 10 -aryl-C 1 ~C 8 -alkoxy, hydroxyl, C O 2 H, C 1 ~C 6 -alkoxycarbonyl, C 6 ~C 10 -aryloxycarbonyl 、C 6 ~C 10 -aryl-C 1 ~C 8 -alkoxycarbonyl, C 1 ~C 6 -alkyl carbonyl, C 6 ~C 10 -arylcarbonyl, C 6 ~C 10 -aryl-C 1 ~C 8 -alkylcarbonyl, C 1 ~C 6 -alkylcarboxy, C 6 ~C 10 -arylcar boxy, C 1 ~C 6 -alkylmercapt yl, C 6 ~C 10 -arylmecapt yl, C 1 ~C 6 -alkylmercaptocarbonyl, C 3 ~C 8 -cycloalkylmercaptocar bonyl, C 6 ~C 10 -arylmecaptocarbonyl, C 1 ~C 6 -alkylmercapt Tocarbonyloxy, C 6 ~C 10 -arylmercaptocarbonyloxy, C 1 ~C 6 -alkylsulfonyl, C ~C 6 ~C 10 -arylsulfonyl, C 1 ~C 6 -alkylsulfinyl, C 6 ~C 10 -arylsulfinyl; each of these is C 1 ~C 6 -alkyl, C 1 ~C 6 -alkoxy, COOH substituted once or several times; CONH 2 , C 1 ~C 6 -alkyl substituted once or twice ; SO 3 H, amino, thiol, hydroxyl, nitro, cyano, fluoro, chloro , bromo, iodo, CF 3 or OCF 3 ; wherein some of these optional substituents are combined to form anellated saturated, unsaturated or aromatic homocyclic or heterocyclic systems; or C 1 ~C 6 -alkyl, C 1 ~C 6 -alkoxy, COOH substituted once or several times forming a saturated, unsaturated or aromatic heterocyclic ring; CONH 2 substituted once or twice.

[0031] In one embodiment, the alkyl chain, alkenyl chain or alkynyl chain is a C 1 ~C 30 chain, preferably a C 1 ~C 18 chain.

[0032] In one embodiment, the alkyl chain, alkenyl chain or alkynyl chain is a C 2 ~C 16 chain, preferably a C 3 ~C 16 chain, more preferably a C 5 ~C 14 chain, even more preferably a C 6 ~C 14 chain.

[0033] In one embodiment, the alkyl chain is a C 5 alkyl chain, a C 6 alkyl chain, a C 7 alkyl chain, a C 8 alkyl chain, a C 9 alkyl chain, a C 10 alkyl chain, a C 11 alkyl chain, a C 12 alkyl chain, a C 13 alkyl chain, or a C 14 alkyl chain.

[0034] In one embodiment, R 1 and R 5 are H.

[0035] In one embodiment, R 2 and R 3 are OH.

[0036] In one embodiment, R 4 is H or OH.

[0037] In one embodiment, R 7 is a C 6 alkyl chain.

[0038] In one embodiment, R 8 and R 9 are independently of each other a C 5 alkyl chain or a C 6 alkyl chain.

[0039] In one embodiment, the halogen is F, Cl, Br, I, or At.

[0040] In one embodiment, the compound is 6-(3,4-dihydroxybenzamide)hexyltri phenylphosphonium bromide.

[0041] In one embodiment, the compound is 6-(3,4,5-trihydroxybenzamide)hexyl triphenylphosphonium bromide.

[0042] In one embodiment, the compound is 5-(6-(3,4,5-trihydroxybenzamide) hexylamino)carbonylpentyl]triphenylphosphonium bromide.

[0043] The present disclosure also relates to any of the compounds disclosed herein for use in medicine, veterinary medicine, or cosmetics, or related compounds.

[0044] In one embodiment, the disclosed compound, or related compound, is used to regulate at least one of the mitochondrial morphology and / or the expression of OXPHOS enzymes. It may be used.

[0045] In one embodiment, the disclosed compound, or related compound, is related to mitochondrial disorders symptoms, or symptoms related to mitochondrial dysfunction in general (including diseases caused by defects in the mitochondrial respiratory chain defects) can be used for the treatment, prevention, or suppression of.

[0046] In one embodiment, mitochondrial disorders include myoclonic epilepsy; red ragged fiber myoclonic epilepsy; Leber's hereditary optic neuropathy; neuropathic ataxia and retinitis pigmentosa; mitochondrial myopathy, encephalopathy, lactic acidosis, stroke; Leigh syndrome; Leigh-like syndrome; dominant optic atrophy; Kearns-Sayre syndrome; maternally inherited diabetes and deafness; Alpers-Huttenlocher syndrome; ataxia neuropathy spectrum; Friedreich's ataxia; chronic progressive external ophthalmoplegia; Pearson syndrome; mitochondrial neurogastrointestinal encephalopathy; Sengers syndrome; neuro-radiological findings of 3-methylglutaconic aciduria, sensorineural deafness, encephalopathy and Leigh-like syndrome; myopathy; mitochondrial myopathy; cardiomyopathy; cerebral myopathy, absence of Leigh syndrome due to cytochrome c oxidase subunit deficiency; isolated or combined OXPHOS deficiencies with hitherto unsolved genetic defects, including disturbances of pyruvate oxidation and ATP + PCR production rates, which are disorders selected from the group consisting of.

[0047] In one embodiment, mitochondrial function incomplete related conditions are , urine tubular acidosis; Parkinson's disease; Alzheimer's disease; amyotrophic lateral sclerosis; Huntington's disease; developmental pervasive disorders; hearing loss; deafness; diabetes; aging; inhibiting mitochondrial function drug are conditions selected from the group consisting of side effects.

[0048] In one embodiment, the compounds of the present disclosure, or related compounds, are used for neurodegenerative diseases, non-alcoholic steatohepatitis, tumors, cancer, kidney disease, scleroderma, hepatic hemosiderosis, hepatic copper overload, alopecia, human pregnancy, acute pancreatitis, fibromyalgia, mitochondrial disorders, or mitochondrial dysfunction or can be used for the treatment or prevention or suppression of conditions related to mitochondrial diseases .

[0049] In one embodiment, the compounds of the present disclosure, or related compounds, are used for neurodegenerative diseases, particularly amyotrophic It can be used for the treatment or prevention of amyotrophic lateral sclerosis.

[0050] In one embodiment, the compounds of the present disclosure, or related compounds, can be used for the treatment or prevention of cancer, wherein the cancer is liver cancer, pancreatic cancer or biliary tract cancer.

[0051] In one embodiment, the compounds of the present disclosure, or related compounds, can be used for the treatment or prevention of non-alcoholic fatty liver disease wherein the non-alcoholic fatty liver disease is non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, or cirrhosis. In one embodiment, the compounds of the present disclosure, or related compounds, can be used for the treatment or prevention of kidney disease,

[0052] wherein the kidney disease is kidney cancer or renal failure. In one embodiment, the tumor disease can be cancer, particularly basal cell carcinoma, bone cancer, intestinal cancer, brain tumor, breast cancer, cervical cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, thyroid cancer or biliary tract cancer.

[0053] In one embodiment, the compounds of the present disclosure, or related compounds, can be used as an antibacterial agent, particularly as a disinfectant. In one embodiment, the compounds of the present disclosure, or related compounds, can be used for the maintenance of pluripotent cell cultures, particularly as a supplement to cell cultures, particularly as a growth medium compound.

[0054] In one embodiment, the present disclosure also relates to a cell culture medium for maintaining pluripotent stem cells in an undifferentiated state, comprising any of the compounds of the present disclosure or related compounds. In one embodiment, the present disclosure also relates to a cell culture medium for maintaining pluripotent stem cells in an undifferentiated state, comprising any of the compounds of the present disclosure or related compounds.

[0055] In one embodiment, the compounds of the present disclosure, or related compounds, can be used for the maintenance of pluripotent cell cultures, particularly as a supplement to cell cultures, particularly as a growth medium compound. In one embodiment, the present disclosure also relates to a cell culture medium for maintaining pluripotent stem cells in an undifferentiated state, comprising any of the compounds of the present disclosure or related compounds.

[0056] The present disclosure also relates to a cell culture medium for maintaining pluripotent stem cells in an undifferentiated state, comprising any of the compounds of the present disclosure or related compounds. In one embodiment, the present disclosure also relates to a cell culture medium for maintaining pluripotent stem cells in an undifferentiated state, comprising any of the compounds of the present disclosure or related compounds.

[0057] In one embodiment, the compounds of the present disclosure, or related compounds, are muscle protectors or body that may be for use as muscle recovery after exercise.

[0058] In one embodiment, the compounds of the present disclosure, or related compounds, are for use in cosmetics, supplements, or nutraceuticals, i.e., as anti-aging agents, or as anti-wrinkle skin care products. that may be for use as.

[0059] In one embodiment, the compounds of the present disclosure, or related compounds, are for use as probes in imaging studies, especially for monitoring mitochondrial imaging studies. that may be for use as.

[0060] The present disclosure also relates to a composition comprising any of the compounds of the present disclosure or related compounds, and one or more pharmaceutically acceptable carriers, adjuvants, excipients, diluents, or combinations thereof, preferably a pharmaceutical composition or a cosmetic composition.

[0061] In one embodiment, the acceptable carrier may be selected from the following list: among others, saline water, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, urea, or combinations thereof.

[0062] In one embodiment, the adjuvant may be selected from the following list: among others, water-in-oil type emulsion adjuvants, aluminum adjuvants, TLR-4 ligands, saponins, and combinations thereof.

[0063] In one embodiment, the excipient may be selected from the following list: among others, glucose, la D - fructose, sucrose, glyceryl monostearate, sodium chloride, glycerol , propylene, glycol, water, ethanol, or combinations thereof.

[0064] In one embodiment, the pharmaceutical composition can be administered, by way of example, orally, parenterally, by inhalation or topically For non - pharmaceutical compositions, i.e., cosmetic compositions, the preferred route is topical .

[0065] In one embodiment, preferred pharmaceutical administration routes include, but are not limited to, oral, parenteral, intramuscular, intravenous, in situ injection, intranasal, sublingual, intratracheal, and inhalation or topical administration .

[0066] In one embodiment, the pharmaceutical composition can be used, for example, in methods for the treatment or prevention of neurodegenerative diseases, non - alcoholic fatty liver disease , tumors, kidney diseases, scleroderma, hepatic hemosiderosis, hepatic copper overload, alopecia, human infertility, acute pancreatitis or fibromyalgia, wherein the pharmaceutical composition is administered in a daily dose .

[0067] In one embodiment, the daily dose of the pharmaceutical composition can be, inter alia, 20 mg / day or 10 mg / day .

[0068] In some embodiments, the dosage or dosage form can be administered to a subject, for example, once a day, twice a day, or three times a day. In other embodiments, the dosage is administered once a week, once a month, once every two months, four times a year, three times a year, twice a year, or once a year .

[0069] In one embodiment, the composition is, in the present subject matter, immunotherapy or any pharmacological approach One or more compounds of the present disclosure, or related compounds, in an amount effective to improve the effectiveness of other therapies containing at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50 %, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, at least 95.7%, at least 98%, or at least 99% of the subject matter may be included.

[0070] In some embodiments, the composition comprises a dose of 0.1 to 1000 mg. For example, in some embodiments, the formulation is 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / k g, 0.4 mg / kg, 0.5 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0. 9 mg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 200 mg / kg, 250 mg / kg, 300 mg / kg, 400 mg / kg, 500 mg / kg, 600 mg / k g, 700 mg / kg, 750 mg / kg, 800 mg / kg, 900 mg / kg, or 1000 mg / kg of the dose. In some embodiments, the composition is 0.1 to 10 mg / kg, 0.1 to 100 mg / kg, 1 to 10 mg / kg, 1 to 100 mg / kg, 1 to 1000 mg / kg, 10 to 100 mg / kg, 10 to 1000 mg / kg, 100 to 1000 mg / kg, 10 to 50 mg / kg, 10 to 25 mg / kg, 10 to 20 mg / kg, 50 to 100 mg / kg, or 100 to 250 mg / kg of dosage.

[0071] The present disclosure also provides nanocarriers, such as liposomes, where the nanocarriers contain the compounds of the present disclosure, or related compounds, or compositions.

[0072] Throughout the detailed description and the claims, the term "comprising" and variations thereof are not intended to exclude other technical features, additives, components, or steps.

[0073] Further objectives, advantages, and features of the solutions disclosed herein will become apparent to those skilled in the art by examining the detailed description, or may be learned by practicing the solutions. .

[0074] Throughout the detailed description and the claims, the term "comprising" and variations thereof are not intended to exclude other technical features, additives, components, or steps. Further objectives, advantages, and features of the solutions disclosed herein will become apparent to those skilled in the art by examining the detailed description, or may be learned by practicing the solutions.

[0075] The following figures provide preferred embodiments for explaining the detailed description and should not be regarded as limiting the scope of the present disclosure.

Brief Description of the Drawings

[0076]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

[0077] Detailed Description

[0078] In one embodiment, and by way of example, a number of hydroxybenzoic acid derivatives (Anti The synthetic strategy pursued for the development of OxBEN) is shown in Figure 1.

[0079] In one embodiment, and by way of example, the mitochondrial tropic antioxidant AntiOxBEN 1 and AntiOxBEN 2 was obtained according to the four-step synthetic strategy shown in Figure 1A. In the example, the starting dimethoxybenzoic acid (1) or trimethoxybenzoic acid (2) is reacted with Bifunctional esters were prepared by amidation reactions using ethyl chloroformate as a coupling agent. The second step reaction was performed by coupling the aryl group to a substituted alkyl spacer (6-aminohexan-1-ol). converts the alcohol functionality (compounds 3 and 4) into a halide, which is a good leaving group. The purpose of the study was to use 1,2-bis(diphenylphosphino)ethane (diphos). The Appel modification reaction afforded the desired compounds (5 or 6) in high yields, especially 70–90%. In the third step, triphenylphosphine (PPh 3SN replaced by 2 Reaction yielded triphenylphosphonium salts (compounds 7 or 8). The hydroxylated analogs (AntiOxBEN 1 and AntiOxBEN 2 ) were synthesized by a demethylation method using boron tribromide (BBr 3 ).

[0080] In one embodiment, and by way of example, the mitochondrial-targeted antioxidant AntiOxBEN 3 was obtained according to the four-step synthetic strategy shown in Figure 1B. Here, trimethoxybenzoic acid (2 ) was coupled to a mono-protected diamine spacer to obtain derivative 9, which was then deprotected in an acidic medium to obtain compound 10. Amine 10 was coupled to the triphenylpho sphonium cationic compound 11 by an amidation reaction. There, an acylating agent was generated in situ. Then, compound 12 was demethylated using a tribromide (BBr 3 ) solution to obtain An tiOxBEN 3 . Overall, moderate yields have been obtained.

[0081] In one embodiment, and by way of example, the antioxidant and redox properties of AntiOxBEN have been reported. Protocatechuic acid and gallic acid were also examined. Vitamin E and trolox were used as standards.

[0082] In one embodiment, the antioxidant ranking activity hierarchy of AntiOxBEN was established by an in vitro non-cellular method. The selected total antioxidant capacity (TAC) assays (DPPH, AB TS and GO) are based on the in situ radical deactivation by antioxidants as a result of radical Spectrophotometric measurements of the decrease in absorbance were performed. Compounds with higher antioxidant activity have lower IC 50 values. The results are shown in Table 1.

[0083] The antioxidant data allows the conclusion that AntiOxBEN is an effective antioxidant and that the I C 50 values followed the same trend in three different assays. From the data obtained, compounds with a pyrogallol moiety, especially AntiOxBEN and AntiOxBEN 2 and AntiOxBEN 3 showed antioxidant activity superior to that of catechol-based, especially AntiOxBEN 1 and can be concluded to be more excellent in antioxidant activity. Generally, the introduction of a triphenylphosphonium (TPP) aliphatic side chain resulted in a slight decrease in antioxidant activity compared to protocatechuic acid and gallic acid.

[0084]

Table 1

[0085] In one embodiment, and by way of example, the redox properties of AntiOxBEN were evaluated (Table 1). The redox potential correlates with the ability of an antioxidant to donate hydrogen atoms and / or electrons to free radicals. Generally, a low oxidation potential (Ep) is associated with excellent antioxidant performance.

[0086] In one embodiment, a glassy carbon working electrode was used to evaluate the oxidation behavior of the parent antioxidants (protocatechuic acid and gallic acid) and AntiOxBEN at physiological pH 7.4 by differential pulse and cyclic voltammetry. The redox data are for protocatechuic Tartaric acid and AntiOxBEN 1 showed a redox potential (Ep ) characteristic of the presence of a catechol group (Ep = 0.257 and 0.224 V, respectively), allowing to conclude so (Table 1). For pyrogallol derivatives (gallic acid, AntiOxBEN 2 , AntiOxB EN 3 ), a significant decrease in the redox potential was observed (Ep = 0.163 - 0.1 68 V) (Table 1).

[0087] In one embodiment, in the differential pulse voltammetry investigation of the mitochondria - targeted antioxidant AntiOxBEN 1 , only one anodic wave was observed. The occurrence of a single voltammetric wave seems to indicate that AntiOxBEN has a lower tendency to adsorb on the electrode surface compared to the parent acid. The differential pulse voltammetry investigations of gallic acid and its derivatives (AntiOxBEN and Anti 1 OxBEN ) revealed the presence of two clearly defined anodic waves at physiological pH. The oxidation peaks are related to the oxidation of the pyrogallol units present in their structures. 2 and Anti OxBEN 3 ) revealed the presence of two clearly defined anodic waves at physiological pH. The oxidation peaks are related to the oxidation of the pyrogallol units present in their structures. 2 anodic waves. The oxidation peaks are related to the oxidation of the pyrogallol units present in their structures. units present in their structures.

[0088] In one embodiment, the cyclic voltammograms obtained for both protocatechuic acid and AntiOxBEN 1 show one anodic peak and a corresponding cathodic peak. The difference between the anodic peak potential value and the cathodic peak potential value indicates an irreversible electron transfer process. The cyclic voltammetry experiment shows a single oxidation peak and no distinct reduction wave is seen in the reverse sweep, for gallic acid and AntiOxBEN and AntiOxBEN showed a single oxidation peak and no distinct reduction wave was seen in the reverse sweep, for gallic acid and AntiOxBEN is seen in the reverse sweep, for gallic acid and AntiOxBEN 2 and AntiOxBEN 3also irreversibly indicates that it has been oxidized.

[0089] In one embodiment, the results obtained show that gallic acid, AntiOxBEN 2 and AntiO xBEN 3 have a lower redox potential than that observed for protocatechuic acid and AntiOxBEN 1 . It is possible to conclude that. The decrease in the oxidation potential is due to the presence of additional phenolic groups in gallic acid and its derivatives (pyrogallol units). It is thought that. The extra hydroxyl groups promote the stabilization of the radical intermediates generated by oxidation, which was converted into a substantial decrease in the obtained redox potential.

[0090] In one embodiment, the introduction of the triphenylphosphonium cation side chain has no notable effect on the redox potential obtained for AntiOxBE N. The data obtained suggest that the structural modifications carried out have a moderate effect on the electron density of the catechol or pyrogallol ring, or no effect at all.

[0091] In one embodiment, the data obtained in the TAC assay are consistent with the AntiOxBEN redox profile . The overall results strengthen the hypothesis that the number of hydroxyl substituents present on the benzoic acid aromatic ring is directly related to the antioxidant properties and the electrochemical properties.

[0092] In one embodiment, and by way of example, the lipophilic properties of AntiOxBEN 1 and AntiOxBEN 2 are evaluated using differential pulse voltammetry (DPV) at physiological pH. Thus. To mimic the movement of ionic drugs across biological membranes, the technique used is often used because this process occurs at the interface between two immiscible electrolyte solutions (ITIES). The ionic drug (C = 0.32 mM), which was initially present in the aqueous phase, transfers to the 1, 6-dichlorohexane (DCH) phase, and the transfer potential (E tr ) is measured by differential pulse voltammetry (DPV). In the ITIES model, as the lipophilicity of the drug increases, the transfer potential (E ) becomes less positive and decreases. tr

[0093] In one embodiment, and by way of example, the transfer potentials (E 1 ) of AntiOxBEN 2 and AntiOxBEN tr were 0.405 V and 0.495 V, respectively. From the data , it was concluded that the presence of additional OH functional groups in AntiOxBEN 2 (a mitochondrial-targeted antioxidant based on gallic acid) translated into an increase in hydrophilicity compared to AntiOxBEN (a mitochondrial-targeted antioxidant based on protocatechuic acid). As expected, hydroxybenzoic acids do not penetrate due to their hydrophilicity 1 . .

[0094] In one embodiment, and by way of example, the chelating properties of AntiOxBEN, i.e., their ability to chelate iron, were determined. Iron is a redox-active metal that can catalyze the Fenton and Haber-Weiss reactions that generate hydroxyl radicals (·OH), which are associated with oxidative damage events that have a profound impact on human health and disease . is a powerful oxidant species. Note that the loss of mitochondrial iron homeostasis and the resulting iron overload can contribute to mitochondrial dysfunction and, thus, different pathologies Therefore, the use of metal chelators or antioxidants that act by this or these mechanisms can function as a therapeutic approach to prevent metal-induced toxicity. Combinations of different mechanisms of action, i.e., by removing harmful reactive species, by hydrogen donation and / or electron transfer, and / or by chelation of pro-oxidant transition metals (i.e., Cu and Fe), phenolic antioxidants that can act may be of utmost significance.

[0095] In one embodiment, the iron (II) chelating ability of the novel AntiOxBEN was evaluated by a ferrozin assay using EDTA (ethylenediaminetetraacetic acid) as a reference. The iron chelating properties of protocatechuic acid, gallic acid, and the mitochondria-targeted antioxidant MitoQ were also evaluated (Figure 2). Since it could completely inhibit the formation of the colored ferrozin-fe(II) complex, EDTA was able to chelate all the iron available in the solution.

[0096] In one embodiment, in contrast to MitoQ, AntiOxBEN (catechol or pyrogallol-based) and hydroxybenzoic acids were able to chelate ferrous iron (Figure 2). Hydroxybenzoic acids (protocatechuic acid and gallic acid) were able to chelate iron efficiently, but those with a pyrogallol moiety were more effective. AntiOxBEN (catechol or pyrogallol- based) was also able to chelate ferrous iron Those with a pyrogallol moiety were more effective. The chelating properties of AntiOxBEN seem to be affected to some extent by the introduction of the TPP cationic spacer when compared to their respective precursors. Still, AntiOxBEN and AntiO 2 xBEN were able to chelate more than 80% of all the iron present in the solution. 3

[0097] In one embodiment, and in the face of the strong antioxidant capacity and iron chelating properties of AntiOxBEN, after the drug discovery optimization program, these innovative antioxidants are predicted to lead to drug candidates that can be applied to suppressing the effects of mitochondrial iron overload and / or reducing mitochondrial iron storage in oxidative stress-related diseases and conditions.

[0098] In one embodiment, and as an example, the mitochondrial uptake of some AntiOxBENs was evaluated in isolated rat liver mitochondria (RLM) in response to the membrane potential. AntiOxBEN can accumulate inside mitochondria driven by ΔΨ (Figure 3A). The profile of different AntiOxBEN accumulations within the mitochondrial matrix was measured. This process was found to be associated with the increase in spacer length and the aromatic substitution pattern (Figure 3B). The small accumulation ratio observed for the pyrogallol derivative AntiOxBEN was significantly improved by the increase in spacer length. The following ranking was achieved: AntiOxBEN 2 was significantly improved by the increase in spacer length. The following ranking was achieved: AntiOxBEN 2 <AntiOxBEN 1 <AntiOxBEN 3All AntiOxBENs show an accumulation rate equivalent to that of MitoQ (Figure 3B).

[0099] The mitochondrial membrane has a high concentration of polyunsaturated fatty acids, which are particularly prone to oxidation because they are located near the ROS generation sites.

[0100] In one embodiment, and by way of example, the anti-oxidation performance of AntiOxBEN against lipid peroxidation of the RLM membrane was determined. Two different oxidative stress agents, FeSO / H 4 / H 2 O 2 / ascorbate and ADP / FeSO 4 were used, as well as two endpoints, namely TBARS generation and oxygen consumption respectively. MitoQ was used as a reference (Figures 4 and 5).

[0101] In one embodiment, gallic acid, AntiOxBEN 4 / H 2 O 2 / ascorbate assay, AntiOxBEN and AntiOxBEN 2 and AntiOxBEN 3 were the most effective mitochondrial-targeted benzoic acid derivatives in preventing mitochondrial lipid peroxidation (Figure 4A). Ant iOxBEN iOxBEN 1 and protocatechuic acid were not effective in preventing TBARS formation in the RLM (Figure 4A). In the ADP / FeSO assay, none of the AntiOxBE 4 N effectively prevented lipid peroxidation (Figures 4B and 5). The ability of AntiOxBEN to suppress lipid peroxidation in the RLM compared to MitoQ decreased in the following order: Mito Q >> AntiOxBEN Q >> AntiOxBEN 3 with gallic acid and AntiOxBEN 2is approximately equal to > Anti OxBEN 1 is approximately equal to protocatechuic acid. Generally, pyrogallol-based AntiO xBEN (Figures 4 and 5) is even more effective in retarding the lipid peroxidation membrane process .

[0102] In one embodiment, and by way of example, the effect of several AntiOxBENs on the mitochondrial permeability transition pore (mPTP) opening was evaluated. Generally, the tested Anti OxBEN had no effect on mPTP opening per se at all tested concentrations.

[0103] In one embodiment, AntiOxBEN 3 was found to cause a concentration-dependent inhibition of calcium-dependent mPTP opening 1 , AntiOxBE N 2 and not MitoQ (Figures 6A - C). This effect is comparable to that of the classical mPTP desensitizer cyclosporin A (1 μM), and may be related to its antioxidant activity or possible chelation of calcium ions. This property can be therapeutically important for preventing and treating graft-versus-host rejection reactions in transplantation, for example, which usually involve mitochondrial destruction in the graft. Since cell metabolism depends on optimal mitochondrial function, the effect of compounds on mitochondrial function parameters can provide information about their toxicity profiles. Thus, by damaging the inner mitochondrial membrane

[0104] or by inhibiting the respiratory chain, ATP synthesis, mitochondrial permeability transition pore (mPTP) process or export machinery, mitochondria ​​Their ability to induce rear malfunction was evaluated.

[0105] In one embodiment, and by way of example, several AntiOxBEN and MitoQ toxic effects on mitochondrial bioenergy, namely RLMΔΨ and respiratory parameters, were measured. ΔΨ represents the main component of the electrochemical gradient generated by mitochondrial respiration and accounts for more than 90% of the total available energy. For mitochondrial respiration assays glutamate / malate (for complex I) and succinate (for complex II) were used as substrates. Furthermore, the mitochondrial oxidative phosphorylation coupling index known as the respiratory control ratio (RCR, state 3 / state 4 respiration) and the ADP / O index (the coupling between ATP synthesis and oxygen consumption) were also calculated. AntiOxBEN and MitoQ were tested at antioxidant-related concentrations, with 10 μM being the highest concentration. ADP / O index (the coupling between ATP synthesis and oxygen consumption) were also calculated. AntiOxBEN and Mi toQ were tested at antioxidant-related concentrations, with 10 μM being the highest concentration.

[0106] In one embodiment, the mitochondrial bioenergy data obtained for MitoQ are shown in Table 2. The results obtained were used for comparative analysis.

[0107]

Table 2

[0108] In one embodiment, it was observed that MitoQ caused a significant decrease in the RCR and ADP / O parameters at all concentrations tested (Table 2). Furthermore, when RLM was incubated with MitoQ concentrations up to 2. 5 μM, using glutamate / malate as a substrate, an increase in state 2, state 4, and oligomycin-inhibited respiration and a decrease in state 3 were observed. were observed. And a decrease in FCCP-uncoupled respiration was observed (Figure 7A). When succinate was used, RLM was completely decoupled in the presence of the highest concentration of MitoQ tested (Figure 7B) . Incubation with increasing concentrations of MitoQ resulted in a progressive decrease in the maximum ΔΨ obtained upon energization (Table 2). Since no repolarization occurred after ADP-induced depolarization, a ΔΨ collapse after ADP addition was observed at 10 μM MitoQ (Table 2).

[0109] In one embodiment, and by way of example, the highest concentration used in the AntiOxBEN toxicity test was such that MitoQ completely disrupted mitochondrial bioenergetics. The data of the AntiOxBEN toxicity test are shown in Tables 3 - 5.

[0110] In one embodiment, and by way of example, AntiOxBEN caused a slight ΔΨ dose-dependent depolarization (10 - 20 mV) after glutamate / malate activation, while promoting a slight overpolarization of 5 - 20 mV under succinate activation. Nevertheless, it is important to note that AntiOxBEN does not have a major impact on RLM ΔΨ.

[0111] [Table 3]

[0112] [Table 4]

[0113] [Table 5]

[0114] ​​In one embodiment, and by way of example, state 2, state 3, state 4, oligomycin-inhibited respiration and mitochondrial respiration assays, and the AntiOxBEN and MitoQ rates for succinate (used as substrate FCCP-stimulated respiration) are shown in FIGS. 7A and B . .

[0115] In one embodiment, it was found that AntiOxBEN induces dose-dependent changes in the respiratory chain. Generally, AntiOxBEN increases state 2, state 4, and oligomycin-inhibited respiration at concentrations higher than 2.5 μM in processes that mainly depend on their lipophilicity and do not rely on their aromatic patterns (catechol vs. pyrogallol). However, it must be emphasized that the observed effect was more evident by using complex I substrates. (FIGS. 7A and B).

[0116] In one embodiment, and by way of example, it was shown that AntiOxBEN induces dose-dependent changes in the respiratory profile of isolated RLM. Some of the observed effects may probably result from the membrane permeabilization effect or proton shuttle activity. This effect can lead to the stimulation of non-linear oxidation respiration and a small ΔΨ depolarization. As a result, AntiOxBEN caused a significant decrease in RCR at all test concentrations. Furthermore, AntiOxBEN (10 μM) also affected the mitochondrial phosphorylation system as evaluated by the change in the ADP / O ratio.

[0117] In one embodiment, and by way of example, the higher concentration observed AntiOxBEN mitochondrial toxicity may be related to the lipophilicity of the spacer and / or the presence of the TPP moiety. ​​​​​​​And even if they were, they would have little relevance to them (catechol vs. pyrogallol). Nonetheless, the presence of the TPP cation and the lipophilic spacer is essential for efficient and sometimes broad mitochondrial accumulation.

[0118] In one embodiment, and by way of example, the mitochondrial-targeted antioxidants, AntiOxBE N and MitoQ, at higher concentrations, cause damage to the inner mitochondrial membrane and thus, or inhibit the respiratory chain, ATP synthesis or export machinery, thereby interfering with mitochondrial respiration.

[0119] In one embodiment, MitoQ effectively inhibits lipid peroxidation in 5 μM RLM (Figs. 4 and 5), but it must be emphasized that it caused toxicity to the mitochondrial bioenergetic apparatus in 2.5 μM RLM (Figs. 7A and B and Table 2).

[0120] In one embodiment, for AntiOxBEN under investigation, it was concluded that RLM toxicity was detected at concentrations higher than those required to exert an antioxidant effect, independently of their mechanism.

[0121] In one embodiment, and by way of example, generally, it was concluded that AntiOxBEN showed a better safety profile than MitoQ.

[0122] In one embodiment, and by way of example, the cytotoxicity of AntiOxBEN was evaluated using monolayer cultures of human hepatocytes (HepG2) derived from hepatocellular carcinoma and the SRB method (Fig. 8). From the data, it was concluded that AntiOxBEN showed low toxicity to HepG2 cells (Fig. 8). AntiOxBEN ​​​​1 promotes slight inhibition of cell proliferation at lower concentrations but stimulates cell proliferation at concentrations higher than 100 μM. Notably, at concentrations lower than 250 μM, AntiOxBEN 2 significantly stimulated cell proliferation, but at concentrations higher than 250 μM, it significantly inhibited cell proliferation. AntiOxBEN 3 significantly inhibited cell proliferation at concentrations higher than 250 μM.

[0123] In one embodiment, and by way of example, it was concluded that the AntiOxBEN toxicity based on its properties (Table 1) and RLM accumulation rate (Figure 3) can be mediated by the lipophilicity of the compound. Generally, AntiOxBEN has a safety margin for HepG2 cells.

[0124] In one embodiment, and by way of example, it was concluded that the structural modifications of benzoic acid (protocatechuic acid and gallic acid) result in a significant improvement in their mitochondrial tropic properties. AntiOxBEN has high antioxidant activity, high mitochondrial accumulation, and low toxicity.

[0125] In one embodiment, the overall results indicate that AntiOxBEN accumulates within mitochondria driven by the mitochondrial membrane potential, prevents lipid peroxidation, and exhibits low intrinsic toxicity. AntiOxBEN shows higher lipophilicity than their parent compounds, such as protocatechuic acid and gallic acid, and

[0126] similar antioxidant and iron chelation properties. In one embodiment, AntiOxBEN is effective against aging and several pathological conditions, such as diabetes, Mitochondrial oxidation related to neurodegenerative diseases including Parkinson's disease and amyotrophic lateral sclerosis It is a mitochondria-targeted antioxidant aimed at preventing or delaying stress.

[0127] In one embodiment, and from the AntiOxBEN series used as an example, pyrogallol Roll-based analogs are potential candidates for the development of a class of drugs with therapeutic uses in mitochondrial oxidation-related disorders. It is predicted to be a potential candidate for the development of a class of drugs.

[0128] Examples of synthetic procedures followed to obtain, as well as several intermediates and AntiOxBEN are provided.

[0129] In one embodiment, the structural characterization of the compound was achieved by spectroscopic analysis. done. 1 H and 13 C spectra The NMR spectra were acquired at room temperature and recorded on a Bruker Avance III operating at 400 and 100 MHz, respectively. Chemical shifts are represented as δ (ppm) values relative to tetramethylsilane (TMS) as an internal standard, and coupling constants (J) are given in Hz. Assignments were also made from DEPT (distortionless e nhancement by polarization transfer) (underlined values). Mass spectra were recorded on a Bruker Microtof (ESI) or Varian 320 -MS (EI) instrument and represented by m / z (% relative) of important fragments.

[0130] In one embodiment, the progress of the reaction was monitored using aluminum silica gel sheets 60 F254 pre-coated with dichloromethane, ethyl acetate, and dichloromethane / methanol in several ratios. -Thin layer chromatography on silica gel 60 (Merck, Darmstadt, Germany) - (TLC) analysis. Spots were detected using UV detection (254 and 366 nm). Flash column chromatography was performed using silica gel 60 (0.040 ~0.063 mm) (Carlo Erba Reactifs - SDS, France ).

[0131] In one embodiment, the acquisition of AntiOxBEN 1 and AntiOxBEN 2 was carried out according to the four-step synthetic strategy shown in Figure 1A. First, intermediate compounds 3 and 4 were synthesized as follows: 3,4-dimethoxybenzoic acid (1) or 3,4,5-trimethoxybenzoic acid ( 2), especially 1 mmol, was dissolved in dichloromethane, especially 40 mL of dichloromethane, and triethylamine, especially 2 mmol of triethylamine, was added. While keeping it in an ice bath, ethyl chloroformate, especially 2 mmol of ethyl chloroformate, was added dropwise to the stirred solution. After stirring at room temperature for 2 hours, the mixture was cooled again, and 6-aminohexan-1-ol, especially 2 mmol of 6-aminohexan-1-ol, was added. The reaction product was stirred at room temperature for 10 hours. The mixture was extracted with dichloromethane, especially 3×20 mL. The organic phases were combined and washed with water, 5% NaHCO 3, especially 20 mL of 5% NaHCO 3 and 1 M HCl, especially 20 mL of 1 M HCl. The combined organic phases were dried and, after filtration, the solvent was evaporated to obtain a white residue. The reaction was monitored by TLC, especially silica gel and ethyl acetate. 3 3 and 1 M HCl, especially 20 mL of 1 M HCl. The combined organic phases were dried and, after filtration, the solvent was evaporated to obtain a white residue. The reaction was monitored by TLC, especially silica gel and ethyl acetate.

[0132] In one embodiment, N-(6-hydroxyhexyl)-3,4-dimethoxybenzamide (3) is characterized as follows. Yield: 74%. 1 H NMR (400 MHz, CDCl 3 ): δ = 1.39 - 1.41 (4H, m, (CH 2 )) 2 (CH 2 )) 2 OH), 1.55 - 1.63 (4H, m, NCH 2 CH 2 (CH 2 )) 2 CH 2 ), 1.99 (1H, s, OH), 3.40 - 3.45 (2H, m, NCH 2 ), 3.63 (2H, t, J = 6.5 Hz, CH 2 OH), 3.91 (6H, s, 2×OCH 3 ), 6.38 (1H, t, J = 5.2 Hz, CONH), 6.85 (1 H, d, J = 8.4 Hz, H(5)), 7.29 (1H, dd, J = 8.4 Hz, J = 2.0 Hz, H(6)), 7.43 (1H, d, J = 2.0 Hz, H(2)). 13 C NMR (100 MHz, CDCl 3 ): δ = 25.4 (CH 2 (CH 2 )) 2 OH), 26.7 (N (CH 2 )) 2 CH 2 ), 29.8 (NCH 2 CH 2 ), 32.6 (CH 2 CH 2 OH), 40.0 (NCH 2 ), 56.1 (2×OCH 3 ), 62.7 ( CH2 OH), 110.4 (C(5)), 110.7 (C(2)), 119.4 (C(6)), 127.5 (C(1)), 149.0 (C(3)), 15 1.7 (C(4)), 167.3 (CONH). EI-MS m / z (%): 281 (M·+), 208 (16), 195 (21), 194 (1 00), 180 (16), 165 (75), 164 (55), 121 (15).

[0133] In one embodiment, the characterization of N-(6-hydroxyhexyl)-3,4,5-trimethoxybenz enzamide (4) is as follows. Yield 82%. 1 H NMR (400 MHz, CDCl 3 ): δ = 1.40-1.43 (4H, m, (CH 2 ) 2 (CH 2 ) 2 OH), 1.54-1.66 (4H, m, NCH 2 CH 2 (CH 2 ) 2 CH 2 ), 1.81 (1H, s, OH), 3.41-3.46 (2H, m, NCH 2 ), 3.64 (2H, t, J = 6.4 Hz, CH 2 OH), 3.87 (3H, s, OCH 3 ), 3.89 (6H, s, 2×OCH 3 ), 6.28 (1H, t, J = 5. 1 Hz, CONH), 7.00 (2H, s, H(2) and H(6)); 13 C NMR (100 MHz, CDCl 3 ): δ = 25.4 ( CH 2 (CH 2 ) 2 OH), 26.7 (NCH2 CH 2 CH 2 ), 29.8 (NCH 2 CH 2 ), 32.6 (CH 2 CH 2 OH), 40.1 (NCH 2 ), 5 6.4 (2×OCH 3 ), 61.0 (OCH 3 ), 62.8 (CH 2 OH), 104.5 (C(2) and C(6)), 130.4 (C(1)) , 140.9 (C(4)), 153.3 (C(3) and C(5)), 167.5 (CONH) and EI-MS m / z (%): 312 (M·+), 225 (38), 224 (34), 211 (59), 196 (49), 195 (100).

[0134] In one embodiment, the general synthetic procedure for obtaining bromohexylbenzamide compounds 5 and 6 is as follows: N-(6-hydroxyhexyl)-3,4-dimethoxybenz amide (3), or N-(6-hydroxyhexyl)-3,4,5-trimethoxybenz amide (4), particularly 1 mmol of hydroxyhexylbenzamide 3, or hydroxy xyhexylbenzamide 4, and 1,2-dibromotetrachloroethane, particularly 1 mm ol of 1,2-dibromotetrachloroethane, are dissolved in THF, particularly 20 mL of THF . After adding 1,2-bis(diphenylphosphino)ethane (diphos), particularly 0.5 mmol, the reaction is stirred particularly at room temperature for 20 hours. Then, the reaction mixture is filtered particularly through a Celite pad. After evaporating the filtrate, an oily residue is obtained. The oil is purified particularly by silica gel flash chromatography using ethyl acetate as the elution system It was carried out. The fraction containing the target compound was collected, the solvent was evaporated, and the product was recrystallized from n-hexane. The reaction was monitored by TLC, especially on silica gel with ethyl acetate.

[0135] In one embodiment, the characterization of N-(6-bromohexyl)-3,4-dimethoxybenzamide (5 ) is as follows. Yield 66%. 1 H NMR (400 MHz, CDCl 3 ): δ = 1.38 - 1.53 (4H, m, (CH 2 )(CH 2 )(CH 2 )Br), 1.59 - 1.67 (2H, 2 m, NCH CH 2 CH 2 ), 1.83 - 1.90 (2H, m, CH 2 CH 2 Br), 3.39 - 3.46 (4H, m, NCH 2 (CH 2 )CH 4 CH 2 Br), 3. 92 (6H, s, 2×OCH 3 ), 6.25 (1H, t, J = 5.4 Hz, CONH), 6.85 (1H, d, J = 8.4 Hz, H( 5)), 7.27 (1H, dd, J = 8.4 Hz, J = 2.0 Hz,, H(6)), 7.43 (1H, d, J = 2.0 Hz, H( 2)); 13 C NMR (100 MHz, CDCl 3 ): δ = 26.2 (NCH 2 CH 2 CH 2 ), 28.0 (CH 2 (CH 2 ) 2 Br), 29.7 (NCH 2 CH 2 ), 32.7 (CH2 CH 2 Br), 33.9 (CH 2 Br), 40.0 (NCH 2 ), 56.1 (OCH 3 ×2), 110.3 (C( 5)), 110.7 (C(2)), 119.2 (C(6)), 127.5 (C(1)), 149.1 (C(3)), 151.7 (C(4)), 167.2 (CONH) and EI-MS m / z (%): 345 (M·+), 343 (24), 264 (36), 195 (34), 194 (19 ), 181 (40), 166 (24), 165 (100).

[0136] In one embodiment, the characterization of N-(6-bromohexyl)-3,4,5-trimethoxybenzamide (6) is as follows. Yield 75%. 1 3 H NMR (400 MHz, CDCl 2 ): δ = 1.37-1.52 (4H, m, (CH 2 ) 2 (CH 2 ) 2 Br), 1.59-1.66 (2H, m, NCH 2 CH 2 ), 1.83-1.90 (2H, m, CH 2 CH 2 Br), 3.39-3.45 (4H, m, NCH 2 (CH 4 ) 2 CH 3 Br), 3. 87 (3H, s, OCH 3 ), 3.88 (6H, s, 2×OCH 13 ), 6.40 (1H, t, J = 5.3 Hz, CONH), 7.01 (2 H, s, H(2) and H(6)); 1313C NMR (100 MHz, CDCl 3 ): δ = 26.2 (NCH 2 CH 2 CH 2 ), 27.9 (CH 2 (CH 2 ) 2 Br), 29.6 (NCH 2 CH 2 ), 32.6 (CH 2 CH 2 Br), 33.9 (CH 2 Br), 40.1 (NCH 2 ), 56.4 (2×OCH 3 ), 61.0 (OCH 3 ), 104.4 (C(2) and C(6)), 130.3 (C(1)), 140.8 (C(4)), 153. 2 (C(3) and C(5)), 167.3 (CONH) and EM / EI m / z (%): 374 (M·+), 372 (15), 22 5 (18), 224 (100), 210 (18), 195 (32), 194 (48).

[0137] In one embodiment, bromohexylbenzamide 5 or 6 (1 mmol l) was mixed with triphenylphosphine (PPh3) (1 mmol) in a round-bottom flask and heated to a temperature of about 12 0 °C for 48 hours. The residue was purified by silica gel flash chromatography using gradient elution (ethyl acetate:methanol). Fractions containing the desired compound were collected and the solvent was evaporated to dryness. The reaction was monitored by TLC (silica gel, ethyl acetate:methanol (9:1) and dichloromethane:methanol (9:1)).

[0138] In one embodiment, 6-(3,4-dimethoxybenzamide)hexyltriphenylphosph ​The characterization of honium bromide (7) is as follows. Yield 65%. 1 H NMR (400 MHz, CD 3 OD): δ = 1.40-1.72 (8H, m, NCH 2 (CH 2 ) 4 ), 3.33-3.37 (2H, m, C H 2 P + Ph 3 ), 3.42-3.49 (2H, m, NCH 2 ), 3.83 (6H, s, 2×OCH 3 ), 6.98 (1H, d, J = 8.5 Hz, H(5)), 7.46 (1H, d, J = 2.1 Hz, H(2)), 7.49 (1H, dd, J = 8.5, J = 2.1 Hz, Hz , H(6)), 7.73-7.89 (15H, m, PPh 3 ); 13 C NMR (100 MHz, CD 3 OD): δ = 22.7 (d, J CP = 51.0 Hz, CH 2 P + Ph 3 ), 23.5 (d, J CP = 4.3 Hz, CH 2 (CH 2 ) 2 P + Ph 3 ), 27.2 (CH 2 (CH 2 ) 3 P + Ph 3 ), 30.3 (NCH 2 CH 2 ), 31.2 (d, J CP = 16.3 Hz, CH 2 CH 2 P + Ph 3 ), 40.8 (NCH 2), 56.7 (2×OCH 3 ), 112.0 (C(5)), 112.2 (C(2)), 120.0 (d, J CP = 86.2 Hz, C(1’)), 122.0 ( C(6)), 128.1 (C(1)), 131.6 (d, J CP = 12.6 Hz, C(3’) and C(5’)), 134.9 (d, J CP = 10.0 Hz, C(2’) and C(6’)), 136.3 (d, J CP = 3.0 Hz, C(4’)), 150.2 (C(3 ))), 153.4 (C(4)), 169.5 (CONH) and EI-MS m / z (%): 511 (M·+), 277 (37), 263 (40), 262 (100), 183 (87), 165 (47), 151 (35), 108 (44), 107 (29), 77 (26), 52 ( 26).

[0139] In one embodiment, the characterization of 6-(3,4,5-trimethoxybenzamide)hexyltriphenyl phosphonium bromide (8) is as follows. Yield 79 %. 1 H NMR (400 MHz, CD 3 OD): δ = 1.41-1.73 (8H, m, NCH 2 (CH 2 )) 4 ), 3.37-3.40 (2H, m, C H 2 P + Ph3), 3.50-3.56 (2H, m, NCH 2 ), 3.94 (3H, s, OCH 3 ), 3.95 (9H, s, 2×OCH 3 ), 7 .28 (2H, s, H(2) and H(6)), 7.75 - 7.90 (15H, m, PPh 3 ); 13 C NMR (100 MHz, CD 3 OD ): δ = 22.5 (d, J CP = 50.8 Hz, CH 2 P + Ph 3 ), 23.3 (d, J CP = 4.0 Hz, CH 2 (CH 2 ) 2 P + P h 3 ), 27.1 (CH 2 (CH 2 ) 3 P + Ph 3 ), 30.0 (NCH 2 CH 2 ), 30.9 (d, J CP = 16.2 Hz, CH 2 CH 2 P + Ph 3 ), 40.6 (NCH 2 ), 57.0 (2×OCH 3 ), 61.1 (OCH 3 ), 106.0 (C(2) and C(6)), 119.7 (d, J CP = 86.1 Hz, C(1’)), 130.8 (C(1)), 131.4 (d, J CP = 12.5 Hz, C(3’) and C(5 ’)), 134.7 (d, J CP = 10.0 Hz, C(2’) and C(6’)), 136.1 (d, J CP = 2.8 Hz, C(4 ’)), 141.6 (C(4)), 154.1 (C(3) and C(5)), 168.7 (CONH) and EI-MS m / z (%): 448 (M·+), 446 (41), 278 (35), 277 (81), 276 (27), 275 (58), 263 (29), 262 (1 00), 185 (31), 184 (25), 183 (94), 152 (21), 108 (36), 96 (53), 94 (54), 77 (24) , 58 (41).

[0140] In one embodiment, triphenylphosphonium salt 7 or 8, particularly 1 mmol of triphenyl phosphonium salt 7, or 1 mmol of triphenylphosphonium salt 8 was dissolved in anhydrous dichloro methane, particularly 15 mL of anhydrous dichloromethane. The reaction mixture was stirred under argon and cooled to a temperature lower than -70 °C. Boron tribromide in dichloromethane, particularly 5 ~7 mmol of 1 M solution of boron tribromide was added to the solution, and the reaction was maintained at particularly -70 °C for 1 0 minutes. After reaching room temperature, the reaction was continued for 12 hours. Then, the reaction was terminated by carefully adding water, particularly 40 m L of water. After removing the water, the resulting product was dissolved in methanol, dried, filtered, and the solvent was evaporated. The residue was purified by silica gel flash chromatography using gradient elution, particularly dichloromethane:meth anol. After collecting the fractions containing the desired compound, the solvent was evaporated to dryness . The reaction was monitored by TLC, particularly silica gel, dichloromethane:methanol (9:1) . The resulting residue was crystallized from ethyl ether / methanol to obtain the corresponding triphenyl phosphonium bromide salt. In one embodiment, 6-(3,4-dihydroxybenzamide)hexyltriphenylpho

[0141] sphonium bromide (AntiOxBEN ​1 )'s structural characterization is as follows . Yield 60%. 1 H NMR (400 MHz, CD 3 OD): δ = 1.35 - 1.47 (2H, m, N(CH 2 ) 4 CH 2 ), 1.50 - 1.75 (6H, m, N CH 2 (CH 2 ) 3 ), 3.33 - 3.47 (4H, m, NCH 2 (CH 2 ) 4 CH 2 P + Ph3), 6.79 (1H, d, J = 8.3 Hz, H(5 )), 7.18 (1H, dd, J = 8.3 Hz, J = 2.2 Hz, H(6)), 7.26 (1H, d, J = 2.2 Hz, H(2)), 7.69 - 7.92 (15H, m, PPh 3 ); 13 C NMR (100 MHz, CD 3 OD): δ = 22.7 (d, J CP = 51.2 Hz , CH 2 P + Ph 3 ), 23.4 (d, J CP = 4.5 Hz, CH 2 (CH 2 ) 2 P + Ph 3 ), 27.0 (CH 2 (CH 2 ) 3 P + Ph 3 ), 3 0.1 (NCH 2 CH 2 ), 31.0 (d, J CP = 16.2 Hz, CH 2 CH 2 P+ Ph 3 ), 40.0 (NCH 2 ), 115.7 (C(5)), 115.8 (C(2)), 120.0 (d, J CP = 86.4 Hz, C(1’)), 120.5 (C(6)), 126.9 (C(1)), 131 .5 (d, J CP = 12.5 Hz, C(3’) and C(5’)), 134.8 (d, J CP = 9.9 Hz, C(2’) and also C(6’)), 136.3 (d, J CP = 3.0 Hz, C(4’)), 146.3 (C(3)), 150.1 (C(4)), 170.3 (C ONH) and ME / ESI m / z (%): 499 (M++H - Br, 51), 498 (M+-Br, 98), 399 (31), 397 (31), 291 (100), 277 (67).

[0142] In one embodiment, the structural characterization of 6-(3,4,5-trihydroxybenzamide)hexyltriphenyl phosphonium bromide (AntiOxBEN 2 ) is as follows: The yield is 50%. 1 2 H NMR (400 MHz, DMSO): δ = 1.23 - 1.50 (8H, m, NCH 2 (CH 2 ) 4 ), 3.11 - 3.16 (2H, m, CH 2 P + Ph3), 3.54 - 3.59 (2H, m, NCH 2 ), 6.81 (2H, s, H(2) and H(6)), 7.74 - 7.91 (15 H, m, PPh 3), 8.00 (1H, t, J = 5.1 Hz, CONH); 13 C NMR (100 MHz, DMSO): δ = 20.2 (d, J CP = 49.8 Hz, CH 2 P + Ph 3 ), 21.8 (d, J CP = 4.1 Hz, CH 2 (CH 2 ) 2 P + Ph 3 ), 25.6 (CH 2 (CH 2 ) 3 P + Ph 3 ), 28.9 (NCH 2 CH 2 ), 29.6 (d, J CP = 16.6 Hz, CH 2 CH 2 P + Ph 3 ), 38.9 (NCH 2 ), 106.7 (C(2) and C(6)), 118.6 (d, J CP = 85.6 Hz, C(1’)), 125.1 (C(1)), 130 .3 (d, J CP = 12.4 Hz, C(3’) and C(5’)), 133.6 (d, J CP = 10.1 Hz, C(2’) and C(6’)), 134.9 (d, J CP = 2.7 Hz, C(4’)), 136.1 (C(4)), 145.4 (C(3) and C (5)), 166.3 (CONH) and ME / ESI m / z (%): 526 (M++Na-Br, 62), 515 (M++H-Br , 30), 514 (M+-Br, 100), 277 (24).

[0143] In one embodiment, AntiOxBEN 3 was carried out according to the four-step synthetic strategy shown in Figure 1B First, 3,4,5-trimethoxybenzoic acid (2) (500 mg, 2.3 mmol) was dissolved in DMF (3.9 mL) at 4 °C, and then N, 2 Cl 2 in (3.9 mL), N, N-diethylpropan-2-amine (0.421 ml, 2.3 mL) and PyBOP( 1668 mg, 2.3 mmol) were added. The mixture was kept in an ice bath and stirred for 0.5 h. Then, tert-butyl (6-aminohexyl) carbamate (0.529 m l, 2.3 mmol) was added and the mixture was warmed to room temperature. The reaction was continued with stirring for 18 h. Next, the mixture was diluted with dichloromethane (20 mL) and washed with saturated NaHCO 3 solution (2 × 10 mL). The organic phase was dried over Na 2 SO 4 filtered, and then concentrated under reduced pressure. The residue was purified by flash chromatography (50% AcOEt / petroleum ether) to give a 73% yield.

[0144] In one embodiment, the structural characterization of the compound tert-butyl (6-(3,4,5-trimethoxybenzamido)hexyl) carbamate) (9) is as follows : 1 H NMR (400 MHz, CDCl 3 ): δ = 7.07 (2H, s, H5, H6), 6.55 (1H, s, H1’), 4.59 (1H , s, H8’), 3.91 (6H, s, 2×OCH 3 ), 3.88 (3H, s, OCH 3), 3.43 (2H, dd, J = 13.0, 6 .9 Hz, H2’), 3.13 (1H, dd, J = 12.6, 6.2 Hz, H7’), 1.67-1.58 (2H, m, H3’), 1. 53-1.32 (15H, m, H4’, H5’, H6’, NHCOOC(CH 3 )); and 13 C NMR (100 MHz, CDCl 3 ) : δ = 167.3 (CONH), 156.3 (NHCOOC(CH 3 )), 153.3 (C3, C5), 140.9 (C4), 130.4 (C1) , 104.5 (C2, C6), 79.3 (NHCOOC(CH 3 )), 61.0 (OCH 3 ), 56.4 (2×OCH 3 ), 40.0 (C7’), 39.7 (C1’), 30.2 (C2’), 29.5 (C6’), 28.5 (NHCOOC(CH 3 )), 26.1 (C3’), 25.8 (C4 ’).

[0145] In one embodiment, the synthesis of N-(6-aminohexyl)-3,4,5-trimethoxybenzamide (10) was as follows: The deprotection step was carried out by adding TFA (4 ml) to a solution of 9 (1 g, 2.4 mmol) in CH 2 Cl 2 (8 ml). The reaction was stirred at room temperature for 1 hour. After neutralization with saturated NaHCO solution, the organic phase was separated. The 3 organic phase was dried over Na SO 2 and filtered, and then concentrated under reduced pressure. The residue was purified by flash 4 chromatography (10% MeOH / CH Cl 2 ). 2) was purified with a 98% yield .

[0146] In one embodiment, the structural characterization of the compound N-(6-aminohexyl)-3,4,5-trimethoxybenz zuamide (10) is as follows. 1 H NMR (400 MHz, MeOD): δ = 7.19 (2H, s, H2, H6), 3.89 (6H, s, 2×OCH 3 ), 3.80 ( 3H, s, OCH 3 ), 3.39 (1H, t, J = 7.1 Hz, H2), 2.99 - 2.90 (2H, m, H7), 1.77 - 1.55 (4H , m, H3, H6), 1.50 - 1.36 (4H, m, H4, H5); and 13 C NMR (100 MHz, MeOD): δ = 169 .4 (CONH), 154.3 (C3, C5), 141.8 (C4’), 131.1 (C1), 105.9 (C2, C6), 61.2 (OCH 3 ) , 56.7 (2×OCH 3 ), 40.8 (C7’), 40.6 (C1’), 30.2 (C6’, 28.4 (C3’), 27.4 (C4’) , 27.0 (C5’).

[0147] In one embodiment, the synthesis of [5-(6-(3,4,5-trimethoxybenzamide)hexylamino)carbonylpentyl]triphenylphosphonium bromide (12) was as follows: To a solution of 10 (689 mg, 2.2 mmol) in DMF (7.4 mL) at 4 °C was added a solution of N,N-diethylpropan-2-amine (0 in CH Cl 2 (7.4 mL) 2 (7.4 mL). .476 mL, 2.7 mmol) and PyBOP (1572 mg, 2.7 mmol) were added. The mixture was kept in an ice bath and stirred for 0.5 h. Then, compound 11 (12 18, 2.7 mmol) was added, and then the reaction was heated to room temperature. The reaction was stirred continuously for 20 3 h. Next, the mixture was diluted with AcOEt (40 mL) and washed with saturated NaHCO 2 solution (2 × 10 mL). The organic phase was dried over Na 4 SO and filtered, and then 2 concentrated under reduced pressure. The residue was purified by flash chromatography (10% MeOH / CH 2 Cl

[0148] In one embodiment, the structural characterization of compound [5-(6-(3,4,5-trimethoxybenzamide)hex ylamino)carbonylpentyl]triphenylphosphonium bromide (12) is as follows. 1 H NMR (400 MHz, CDCl 3 ): δ = 7.85 - 7.76 (3H, m, H4”), 7.73 - 7.59 (12H, m, H2”, H3”, H5”, H6”), 7.12 (2H, s, H2, H6), 6.93 (1H, t, J = 5.7 Hz, H1’), 6.26 (1H , t, J = 5.7 Hz, H8’), 3.88 (6H, s, 2×OCH 3 ), 3.85 (1H, s, OCH 3 ), 3.39 (dd, J = 13.2, 6.7 Hz, 1H), 3.19 - 3.05 (4H, m, H7’, H14’), 2.14 (1H, t, J = 7.1 Hz, H10 ’), 1.69 - 1.26 (14H, m, H3’, H4’, H5’, H6’ H11’, H12’, H13’); and 13 C NMR (100 MHz, CDCl 3 ): δ = 173.3 (C9’), 167.2 (PhCONH), 153.1 (C3, C5), 140.4 ( C4), 135.4 (d, J CP = 2.9 Hz, C4”), 133.4 (d, J CP = 9.9 Hz, C2”,C6”), 130.7 (d , J CP = 12.6 Hz, C3”,C5”), 130.4 (C1), 117.9 (d, J CP = 86.2 Hz, C1”), 104.5 ( C2, C6), 60.9 (OCH 3 ), 56.4 (2×OCH 3 ), 39.7 (C2’), 39.0 (C7’), 36.3 (C10’), 30 .0 (C3’), 29.8 (C6’), 28.9 (d, J = 5.0 Hz, C12’), 26.6 (C4’), 25.9 (C5’), 2 4.9 (C11’), 22.3 (d, J = 43.5 Hz, C14’), 22.1 (d, J = 12.5 Hz, C13’).

[0149] In one embodiment, [5-(6-(3,4,5-trihydroxybenzamide)hexyl amino)carbonylpentyl]triphenylphosphonium bromide (AntiOxBEN 3 ) was synthesized as follows. 1.0 g, 1.4 mmol was dissolved in 7.6 ml of anhydrous dichloromethane . The reaction mixture was stirred under argon and cooled to a temperature lower than -75 °C . Boron tribromide (4.3 ml of a 1 M solution in dichloromethane; 4.3 mmol) solution was added dropwise and the reaction was maintained at -75 °C for 10 minutes. Once the addition was complete, the reaction was held at -7 0 °C for 10 minutes and then warmed to room temperature with continuous stirring for 12 hours. Thereafter, water (20 mL) was carefully added to terminate the reaction. After removing the water, the resulting product was dissolved in methanol and dried over anhydrous Na 2 SO 4 and filtered, and the solvent was evaporated. The residue was purified by flash chromatography (10% MeOH / CH 2 Cl 2 ) to give a 55% yield.

[0150] In one embodiment, the structural characterization of the compound [5-(6-(3,4,5-trihydroxybenzamide)he xylamino)carbonylpentyl]triphenylphosphonium bromide (AntiOx BEN 3 ) is as follows. 1 H NMR (400 MHz, MeOD): δ = 7.92 - 7.83 (3H, m, H4”), 7.82 - 7.70 (12H, m, 12H, m, H2”, H3”, H5”, H6”), 6.83 (2H, s, H2, H6), 3.43 - 3.34 (2H, m, H14’), 3.33 - 3.25 (2H, m, H2’), 3.14 (1H, t, J = 6.9 Hz, H7’), 2.15 (1H, t, J = 7.0 Hz, H1 0’), 1.72 - 1.28 (14H, m, H3’, H4’, H5’, H6’ H11’, H12’, H13’); 13 C NMR (1 00 MHz, MeOD): δ = 176.3 (C9), 170.5 (PhCOONH), 146.5 (C3, C5), 138.1(C4), 136. 1 (d, J = 2.9 Hz, C4”), 134.7 (d, J CP = 10.0 Hz, C2”, C6”), 131.5 (d, J CP = 12 .6 Hz, C3”, C4”), 125.4 (C1), 119.7 (d, J CP = 86.3 Hz, C1”), 107.8 (C2, C6), 40.8 (C2’), 40.5 (C7’), 36.2 (C10’), 30.9 (C3’), 30.8 (C6’), 30.2 (C4’), 2 9.9 (C5’), 27.4 (d, J CP = 2.5 Hz, C12’), 26.1 (C11’), 23.1 (d, J CP = 4.2 Hz, C13’), 22.6 (d, J CP = 51.3 Hz, C14’); ESI / MS m / z (%): 628 (M + +H - Br - , 38), 627 (M + - Br, 100), 556 (35), 547 (46); and ESI / HRMS m / z calculated value C 37 H 44 N 2 O 5 P + (M + - Br - ): 627.2982; measured value 627.2970.

[0151] The radical scavenging activity of AntiOxCIN was evaluated by total antioxidant capacity assays based on DPPH · , ABTS ·+ and GO · radicals. All of these methods are based on the decrease in absorbance due to the inactivation of radicals (DPPH by antioxidants (DPPH · , ABTS ·+ or GO · ). The results are shown in IC 50 This is expressed as a 50% reduction in the amount of radicals. The antioxidant assay was performed at BioTech. A multiplate reader (Powerwave XS) manufactured by H Instruments The data were collected using a Microplate Reader.

[0152] In one embodiment, DPPH radical scavenging activity was performed as follows: Solutions of test compounds (ranging from 0 μM to 500 μM) were prepared in ethanol. An ethanol solution (6.85 mM) was also prepared and then measured for an absorbance of 0.72 ± 0.02 at 515 nm. Each compound solution (20 μL) was diluted with 180 μL of DPPH· solution. The absorbance at 515 nm was recorded every minute for 45 minutes. The toxicity was measured using a blank (20 μL ethanol and 180 μL ethanol equivalent to 100% radicals) The IC was based on a comparison of the IC10 concentration of DPPH· in the presence of the test compound. 50 Dosage for determination of value Response curves were established and data are the mean ± SEM of three independent experiments.

[0153] In one embodiment, ABTS ·+ The scavenging activity was assessed as follows: A solution of the test compound in ethanol was prepared at 150 mM persulfate (range: 1 μM to 500 μM). A solution of potassium phosphate (163 μL) was added to 10 mL of 7 mM aqueous ABTS solution, followed by ABTS by storing in the dark at room temperature for 16 hours · Radical cation solution (2.45 The solution was then diluted with ethanol to obtain an absorbance of 0.72 ± 0.05 mM. Compounds (20 μL) were added in triplicate to ABTS·+ After adding to the solution (180 μL), spectrophotometric measurements were performed every minute over 15 minutes. The inhibition rate of radicals was based on the comparison between the blank corresponding to 100% radicals (20 μL of ethanol and 180 μL of ABTS solution ) and the test compound solution. A dose - response curve was established for the determination of the IC ·+ value. The data are the mean ± SEM of three independent experiments. ) and the test compound solution. IC 50 value. The data are the mean ± SEM of three independent experiments. value. The data are the mean ± SEM of three independent experiments.

[0154] In one embodiment, the GO · scavenging activity was evaluated as follows: Solutions of test compounds at concentrations of 10 μM - 100 μM were prepared in ethanol. An ethanol solution of 5 mM GO was prepared and diluted to reach an absorbance of 1.00 ± 0.02 at 428 nm. Following the addition of three consecutive compound solutions (20 μL) to the GO· solution ( · 180 μL), absorbance measurements at 428 nm were performed in the dark at room temperature over 30 minutes. The inhibition rate of radicals was based on the comparison between the blank corresponding to 100% radicals (20 μL of ethanol and 180 μL of GO solution) and the test compound solution. A dose - response curve was established for the determination of the IC value. The data are the mean ± SEM of three independent experiments. value. The data are the mean ± SEM of three independent experiments. solution) and the test compound solution. IC · value. The data are the mean ± SEM of three independent experiments. value. The data are the mean ± SEM of three independent experiments. 50 value. The data are the mean ± SEM of three independent experiments. value. The data are the mean ± SEM of three independent experiments.

[0155] In one embodiment, the redox and lipophilic properties of AntiOxBEN were evaluated by electrochemistry techniques.

[0156] In one embodiment, the electrochemistry analysis data were obtained using a computer - controlled potentiostat Aut olab PGSTAT302N (Metrohm Autolab, Utrecht, The Netherlands) It was obtained using . Generally, cyclic voltammetry (CV) data was acquired at a scan rate of 50 mVs -1 . The differential pulse voltammetry (DPV) results were obtained at a step potential of 4 mV, a pulse amplitude of 50 mV, and a scan rate of 8 mVs -1 . Electrochemical signals were monitored using the general-purpose electrochemical system (GPES) version 4.9, software package . To minimize the contribution of background noise to the analytical signal, all electrochemical experiments were conducted in an electrochemical cell placed inside a Faraday cage at room temperature .

[0157] In one embodiment, the evaluation process of the redox properties of AntiOxBEN was carried out as follows : The stock solution (10 mM) of each compound was prepared by dissolving an appropriate amount in ethanol . The voltammetry working solution was prepared in the electrochemical cell at a final concentration of 0.1 mM . The supporting electrolyte at pH 7.4 was prepared by diluting 6.2 mL of 0.2 M dipotassium hydrogen phosphate and 43 .8 mL of 0.2 M potassium dihydrogen phosphate to 100 mL . Voltammetry data was obtained in a three-electrode system consisting of a glassy carbon electrode (GCE, d = 2 mm ) as the working electrode, a platinum wire counter electrode, and a saturated Ag / AgCl reference electrode . In one embodiment, the evaluation of the lipophilic properties of AntiOxBEN was carried out as follows . The electrochemical cell was a four-electrode system having an array of micro liquid-liquid interfaces (μITIES) containing two Ag / AgCl reference electrodes and two Pt counter electrodes in each phase. The microporous membrane was placed on a glass cylinder filled with 4.0 mL of the aqueous phase with a fluorosilicone seal Sealed with Rant (Dow Corning 730), and an aliquot of the AntiOxBEN solution was added thereto. Next, the membrane was immersed in the organic phase contained in the cell. The organic phase reference solution (2 mM BTPPAC1 + 2 mM aqueous NaCl solution) was mechanically stabilized. The aqueous supporting electrolyte solution was 10 mM Tris-HCl buffer at pH 7.0. (2 mM of BTPPAC1 + 2 mM aqueous solution of NaCl) was mechanically stabilized. The aqueous supporting electrolyte solution was 10 mM Tris-HCl buffer at pH 7.0.

[0158] In one embodiment, the AntiOxBEN iron chelating properties were evaluated by the spectrophotometric ferrozine method performed with a multi-plate reader (Powerwave XS Microplate Reader) from Bio-Tech instruments. truments' multi-plate reader (Powerwave XS Microp late Reader).

[0159] In one embodiment, the AntiOxBEN iron chelating properties were evaluated as follows: To each well, a solution of the test compound (100 μM) and ammonium iron(II) sulfate in ammonium acetate (20 μM) was added, incubated for 10 minutes, and its absorbance was read at 562 nm. To each well, a solution of the test compound (100 μM) and ammonium iron(II) sulfate in ammonium acetate (20 μM) was added, incubated for 10 minutes, and its absorbance was read at 562 nm. Next, freshly prepared ferrozine solution (5 mM) was added to each well (final concentration of 96 μM). After a new incubation at 37 °C for 10 minutes, the absorbance of the [Fe(ferrozine)3] complex was measured at 562 nm. Blank wells were tested using DMSO instead of the test compound. EDTA was used as a reference. All compounds were tested at a final concentration of 100 μM. The absorbance of the first measurement was subtracted to the final value to eliminate the absorbance by the test compound. The data are the mean ± SEM of 3 independent experiments and are expressed as % of Fe(II) chelation (EDTA = 100%). 6 μM final concentration). After a new 10-minute incubation at 37 °C, the absorbance of the [Fe(ferrozine)3] complex was measured at 562 nm. Blank wells were tested using DMSO instead of the test compound. EDTA was used as a reference. All compounds were tested at a final concentration of 100 μM. The absorbance of the first measurement was subtracted to the final value to eliminate the absorbance by the test compound. The data are the mean ± SEM of 3 independent experiments and are expressed as % of Fe(II) chelation (EDTA = 100%). 2+ complex was measured at 562 nm. Blank wells were tested using DMSO instead of the test compound. EDTA was used as a reference. All compounds were tested at a final concentration of 100 μM. The absorbance of the first measurement was subtracted to the final value to eliminate the absorbance by the test compound. The data are the mean ± SEM of 3 independent experiments and are expressed as % of Fe(II) chelation (EDTA = 100%). was used to test the blank wells. EDTA was used as a reference. All compounds were tested at a final concentration of 100 μM. The absorbance of the first measurement was subtracted to the final value to eliminate the absorbance by the test compound. The data are the mean ± SEM of 3 independent experiments and are expressed as % of Fe(II) chelation (EDTA = 100%). The absorbance of the first measurement was subtracted to the final value to eliminate the absorbance by the test compound. The data are the mean ± SEM of 3 independent experiments and are expressed as % of Fe(II) chelation (EDTA = 100%). The absorbance of the first measurement was subtracted to the final value to eliminate the absorbance by the test compound. The data are the mean ± SEM of 3 independent experiments and are expressed as % of Fe(II) chelation (EDTA = 100%). (EDTA = 100%).

[0160] In one embodiment, the evaluation of the AntiOxBEN functional mitochondrial toxicity profile was performed in rat liver mitochondria (RLM). The RLM was prepared by tissue homogenization and subsequent fractionation by centrifugation in an ice-cold buffer containing 250 mM sucrose, 10 mM HEPES (pH 7.4), 1 mM EGTA, and 0.1% fat-free bovine serum albumin. After obtaining the crude mitochondrial preparation, the pellet was washed twice and resuspended in a wash buffer (250 mM sucrose and 10 mM HEPES, pH 7.4). The protein concentration was determined by the biuret assay using BSA as a standard.

[0161] In one embodiment, the AntiOxBEN uptake by mitochondria was evaluated.

[0162] In one embodiment, the uptake of AntiOxBEN by activated RLM was evaluated as follows: RLM (0.5 mg protein / mL) was incubated with AntiOxBEN at 37 °C in 1 mL of KCl medium (120 mM KCl, 10 μL HEPES, pH 7.2 and 1 mM EGTA). Five consecutive 1 μM additions of each AntiOxBEN were performed to calibrate the electrode response in the presence of rotenone (1.5 μM). Subsequently, succinate (10 mM) was added to generate ΔΨ. At the end of the assay, valinomycin (0.2 μg / mL) was added to dissipate ΔΨ. The measurements were performed using an ion-selective electrode that measures the distribution of tetraphenylphosphonium cation (TPP + ) and a reference Ag / AgCl electrode. The amount accumulated within the mitochondria was determined within the mitochondria 2 When assuming a capacity of 0.5 μL / mg protein, the disappearance of AntiOxBEN from the mitochondrial outer medium to the mitochondrial inner medium and the mitochondrial uptake of the TPP compound were calculated by binding correction.

[0163] The results of AntiOxBEN against RLM lipid peroxidation were evaluated. Two different methods were used.

[0164] In one embodiment, the effect of AntiOxBEN against RLM lipid peroxidation was measured by the following thiobarbituric acid reactive species (TBARS) assay: RLM (2 mg protein / ml) was incubated at 37 °C in 0.8 mL of medium containing 100 mM KCl, 10 mM Tris-HCl, pH 7.6 supplemented with 5 mM glutamate / 2.5 mM phosphate as substrate. RLM was incubated with each AntiOxBEN (5 μM) for 5 minutes, and then mitochondria were exposed to oxidative stress conditions by adding 100 μM FeSO / 500 μM H 4 / 5 mM ascorbate at 37 °C for 15 minutes. After exposure to oxidative stress, 60 μL of 2% (v / v) 2 butylated hydroxytoluene in DMSO was added, followed by 200 μL of 35% (v / v) perchloric acid and 200 μL 2 of 1% (w / v) thiobarbituric acid. The sample was then incubated at 10 0 °C for 15 minutes, cooled, and its supernatant was transferred to a glass tube. After adding 2 mL of MilliQ water and 2 mL of butan-1-ol, the sample was vortexed vigorously for several seconds. The two phases were separated. The fluorescence of an aliquot (250 μL) of the organic layer was measured for TBARS. ​​​​Subsequently, it was analyzed using a plate reader (λEx = 515 nm; λEm = 553 nm). Glu The TBARS background production in RLM activated with glutamate / glutarate was found to be negligible as much as possible. The data are the mean ± SEM of three independent experiments and are expressed as a percentage of the control (control = 100%).

[0165] In one embodiment, the effect of AntiOxBEN on RLM lipid peroxidation was measured by a second methodology as follows : The oxygen consumption of 2 mg of RLM in a total volume of 1 mL of reaction medium consisting of glutamate / glutarate (5 mM / 2.5 mM) as respiratory substrate, 100 mM KCl, 10 mM Tris-HCl and pH 7.6 was monitored at 37 °C using a Clark oxygen electrode. The RLM was incubated with each AntiOxBEN (5 μM) for 5 minutes, and then the lipid peroxidation process was initiated by adding 1 mM of ADP and 0.1 mM of FeSO (final concentration) 4 . The oxygen consumption over time due to the peroxidation of the RLM membrane by the oxidation promoter pair (1 mM ADP 2 / 0.1 mM FeSO 4 ) was recorded. The traces are the mean ± SEM recordings from six independent experiments. The effectiveness of AntiOxBEN in preventing lipid peroxidation was measured using the time lag phase associated with the slower oxygen consumption following the addition of ADP ) / Fe . The data are the mean ± SEM from six independent 2+ experiments and are expressed as a percentage of the control (control = 100%).

[0166] In one embodiment, the effect of AntiOxBEN on mitochondrial permeability transition pore opening was evaluated. The result was evaluated.

[0167] In one embodiment, the effect of AntiOxBEN on mitochondrial permeability transition pore opening was evaluated. The result was measured as follows. Mitochondrial swelling was estimated by measuring the change in light scattered from the mitochondrial suspension when monitored spectrophotometrically at 540 nm. Increasing concentrations of AntiOxBEN (2.5 - 10 μM) were added to a reaction medium (200 mM sucrose, 1 mM KH PO , 10 mM Tris (pH 7 2 PO 4 , 10 mM Tris (pH 7 .4), 5 mM succinate, and 1.5 μM rotenone added to 10 μM EGTA) in the presence of RLM (1 mg) and incubated for 5 minutes before the assay. The experiment was initiated by adding appropriate titrated daily concentrations of Ca (15 - 50 μM). Cyclosporin A (CsA), a PTP desensitizer, was added to demonstrate mPTP opening. The reaction was continuously stirred and the temperature was maintained at 37 °C. The data are the mean ± SEM of three independent experiments and are expressed as the Δ absorbance at 540 nm. In one embodiment, the effect of AntiOxCIN on mitochondrial respiration was evaluated. 2+ (15 - 50 μM). Cyclosporin A (CsA), a PTP desensitizer, was added to demonstrate mPTP opening. The reaction was continuously stirred and the temperature was maintained at 37 °C. The data are the mean ± SEM of three independent experiments and are expressed as the Δ absorbance at 540 nm. In one embodiment, the effect of AntiOxCIN on mitochondrial respiration was evaluated. In one embodiment, the evaluation of the AntiOxBEN effect on mitochondrial respiration was performed as follows: The respiration of isolated RLM was evaluated polarographically using a Clark-type oxygen electrode connected to an appropriate recorder in a 1 mL thermostatted water-jacketed chamber with magnetic stirring at 37 °C. The standard respiration medium was 130 mM sucrose, 50 mM KCl, 5 In one embodiment, the effect of AntiOxBEN on mitochondrial respiration was evaluated.

[0168] In one embodiment, the effect of AntiOxCIN on mitochondrial respiration was evaluated.

[0169] In one embodiment, the evaluation of the AntiOxBEN effect on mitochondrial respiration was performed as follows: The respiration of isolated RLM was evaluated polarographically using a Clark-type oxygen electrode connected to an appropriate recorder in a 1 mL thermostatted water-jacketed chamber with magnetic stirring at 37 °C. The standard respiration medium was 130 mM sucrose, 50 mM KCl, 5 In one embodiment, the evaluation of the AntiOxBEN effect on mitochondrial respiration was performed as follows: The respiration of isolated RLM was evaluated polarographically using a Clark-type oxygen electrode connected to an appropriate recorder in a 1 mL thermostatted water-jacketed chamber with magnetic stirring at 37 °C. The standard respiration medium was 130 mM sucrose, 50 mM KCl, 5 In one embodiment, the evaluation of the AntiOxBEN effect on mitochondrial respiration was performed as follows: The respiration of isolated RLM was evaluated polarographically using a Clark-type oxygen electrode connected to an appropriate recorder in a 1 mL thermostatted water-jacketed chamber with magnetic stirring at 37 °C. The standard respiration medium was 130 mM sucrose, 50 mM KCl, 5 In one embodiment, the evaluation of the AntiOxBEN effect on mitochondrial respiration was performed as follows: The respiration of isolated RLM was evaluated polarographically using a Clark-type oxygen electrode connected to an appropriate recorder in a 1 mL thermostatted water-jacketed chamber with magnetic stirring at 37 °C. The standard respiration medium was 130 mM sucrose, 50 mM KCl, 5 mM KH 2 PO 4 、 5 mM HEPES (pH 7.3) and 10 μM EGTA to make. Increasing concentrations of AntiOxBEN (2.5 - 10 μM) were added to the reaction medium containing the respiratory substrates glutamate / malate (10 mM and 5 mM respectively) or succinate (5 mM) and RLM (1 mg), and incubated for 5 minutes before the assay. State 2 was considered as the respiration during the 5 - minute incubation with AntiOxBEN To induce state 3 respiration, 125 nmol of ADP (using glutamate / malate ) or 75 nmol of ADP (using succinate) was added. State 4 was determined after the completion of ADP phosphorylation. The subsequent addition of oligomycin (2 μg / ml) inhibited ATP synthase and was derived from the oligomycin - inhibited respiratory state. Finally, 1 μM of FCCP was added to induce uncoupled respiration. RCR was 7.3 ± 0. 6 and 4.1 ± 0.3 for the control experiments using glutamate - malate or succinate respectively as the respiratory substrate. With the same respiratory substrate, the ADP / O ratio was 2.6 ± 0.1 and 1.5 ± 0.1 respectively. The data are the mean ± SEM of 7 independent experiments .

[0170] In one embodiment, the effect of AntiOxBEN on the transmembrane potential (ΔΨ) was evaluated .

[0171] In one embodiment, the evaluation of the AntiOxBEN effect on the mitochondrial transmembrane potential (ΔΨ) was performed as follows: The mitochondrial transmembrane potential (ΔΨ) was determined using safranine . ​Estimated through evaluation of fluorescence changes at 5 μM, and recorded on a spectrofluorometer operating at an excitation wavelength of 495 and an emission wavelength of 586 nm with a slit width of 5 nm. Increasing concentrations of AntiOxB EN (2.5 - 10 μM) were added to reaction media (200 mM sucrose, 1 mM KH PO 4, 10 mM Tris(pH 7.4) and 10 μM EGTA) containing the respiratory substrates glutamate / malate (5 mM and 2.5 mM respectively) or succinate (5 mM) and RLM (0.5 mg in a final volume of 2 mL ) and incubated for 5 minutes at 25 °C prior to assay. In this assay, safranine (5 μM) and ADP (25 nmol 2 PO 4 4, 10 mM Tris(pH 7.4) and 10 μM EGTA) containing the respiratory substrates glutamate / malate (5 mM and 2.5 mM respectively) or succinate (5 mM) and RLM (0.5 mg in a final volume of 2 mL ) were used at the start and to induce depolarization of the assay respectively. Subsequently, 1 μM FCCP was added at the end of all experiments to depolarize the mitochondria . ΔΨ was calculated using a calibration curve obtained when RLM was incubated in a K -free reaction medium containing 200 mM sucrose, 1 mM NaH PO 4, 10 mM Tris(pH 7.4) and 10 μM EGTA supplemented with 0.4 μg valinomycin. The amplification of the fluorescence change of safranine induced by ΔΨ was found to be similar in standard and K 2 PO 4 4, 10 mM T ris(pH 7.4) and 10 μM EGTA supplemented with 0.4 μg valinomycin. The amplification of the fluorescence change of safranine induced by ΔΨ was found to be similar in standard and K + -free media . "Repolarization" corresponded to the recovery of the membrane potential after complete phosphorylation of the added ADP. The delay phase reflected the time required to phosphorylate the added ADP . When glutamate / malate or succinate were used respectively, isolated RLM had a ΔΨ approximately equal to 226 mV and a ΔΨ (negative inside) approximately equal to 202 mV + . Similar results were found. "Repolarization" corresponded to the recovery of the membrane potential after complete phosphorylation of the added ADP. The delay phase reflected the time required to phosphorylate the added ADP . When glutamate / malate or succinate were used respectively, isolated RLM had a ΔΨ approximately equal to 226 mV and a ΔΨ (negative inside) approximately equal to 202 mV . The isolated RLM had a ΔΨ approximately equal to 226 mV and a ΔΨ (negative inside) approximately equal to 202 mV occurred. The data are the mean ± SEM of 5 independent experiments.

[0172] In one embodiment, the cytotoxicity profile of AntiOxBEN was evaluated in human hepatocellular carcinoma He pG2 cells. Human hepatocellular carcinoma HepG2 cells were cultured in high glucose medium consisting of Dulbecco's Modified Eagle Medium (DMEM; D5648) supplemented with sodium pyruvate ( 0.11 g / L), sodium bicarbonate (1.8 g / L), and 10% fetal bovine serum (F BS) and 1% antibiotic penicillin-streptomycin 100× solution. The cells were maintained at 37 °C in a humidified incubator with 5% CO . HepG2 cells 2 were seeded at a density of 4 × 10 cells / mL and allowed to grow for 24 hours prior to treatment. 4

[0173] In one embodiment, the cytotoxicity screening was performed as follows: The cells were placed in 48-well plates (2 × 10 4 cells / 500 μL) and then incubated with AntiOxBEN at concentrations ranging from 25 μM to 500 μM for 48 hours. After incubation, the sulforhodamine B (SRB) assay was used for cell density determination based on the measurement of cell protein content. Briefly, after incubation, the medium was removed and the wells were rinsed with PBS (1×). The cells were fixed by adding 1% acetic acid in 100% methanol at -20 °C for at least 2 hours. Then, the fixative was discarded and the plates were dried in an oven at 37 °C. 250 microliters of 0.5% SRB in 1% acetic acid solution was added and incubated at 37 °C for 1 hour. Then the wells were washed with 1% acetic acid in water and the plates were dried in an oven at 37 °C. The fixative was discarded and the plates were dried in an oven at 37 °C. 250 microliters of 0.5% SRB in 1% acetic acid solution was added and incubated at 37 °C for 1 hour. Then the wells were washed with 1% acetic acid in water Washed and dried. Then, 500 μl of Tris (pH 10) was added and the plate was stirred for 15 minutes. Finally, 200 μl of each supernatant was transferred to a 96-well plate and the optical density was measured at 540 nm. The data are the mean ± SEM of four independent experiments, and the results are expressed as the percentage of the control (control = 100%), which represents the cell density without any treatment at each time point.

[0174] In one embodiment, all biological data were analyzed as follows: using GraphPad Prism 5.0 software (GraphPad Software, Inc.) to represent all results as the mean ± SEM relative to the number of experiments shown. The data were analyzed by Student's t-test for comparison of two means and one-way ANOVA with Dunnett's multiple comparison post hoc test. The last test was used to compare more than two groups with one independent variable. Significance was determined at * P < 0.05, ** P < 0.01, *** P < 0.0005, **** P < 0.0001.

[0175] The present disclosure should not be considered to be in any way limited to the described embodiments, and those skilled in the art will foresee many possibilities for modification.

[0176] The above-described embodiments can be combined. The appended claims further illustrate specific embodiments of the present disclosure.

Prior Art Documents

Non-Patent Documents

[0177]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Claims

1. The following compound for use in medicine: 【Chemical Formula 1】 ; or [Chemical 2] or a salt, solvate, hydrate, tautomer, stereoisomer. (wherein, R １ and R ５ are H; R ２ and R ３ is OH; R ４ is H or OH; R ７ is a C ６ alkyl chain; Z is F, Cl, Br, I or At. )

2. The compound according to claim 1, wherein Z is Cl or Br.

3. The compound according to any one of claims 1 or 2, wherein the compound is 6-(3,4-dihydroxybenzamide)hexyltriphenylphosphonium bromide, or 6-(3,4,5-trihydroxybenzamide)hexyltriphenylphosphonium bromide.

4. The compound according to any one of claims 1 to 3 for use in the prevention or suppression of symptoms associated with mitochondrial disorders or symptoms associated with mitochondrial dysfunction or conditions associated with mitochondrial diseases.

5. The mitochondrial disorder is myoclonic epilepsy; red ragged fiber myoclonic epilepsy; Leber's hereditary optic neuropathy; neuropathic ataxia and retinitis pigmentosa; mitochondrial myopathy, encephalopathy, lactic acidosis, stroke; Leigh syndrome; Leigh-like syndrome; dominant optic atrophy; Kearns-Sayre syndrome; maternally inherited diabetes and deafness; Alpers-Huttenlocher syndrome; ataxia neuropathy spectrum; Friedreich's ataxia; chronic progressive external ophthalmoplegia; Pearson syndrome; mitochondrial neurogastrointestinal encephalopathy; Sengers syndrome; 3-methylglutaconic aciduria, sensorineural deafness, encephalopathy and neurological radiological findings of Leigh-like syndrome; myopathy; mitochondrial myopathy; cardiomyopathy; cerebral myopathy, absence of Leigh syndrome due to deficiency of surfactant protein of complex IV; disorders selected from the group consisting of isolated or combined OXPHOS deficiency with hitherto unsolved genetic defects including disturbances in pyruvate oxidation and ATP + PCR production rate; or The condition associated with the mitochondrial dysfunction is a condition selected from the group consisting of tubular acidosis; Parkinson's disease; Alzheimer's disease; amyotrophic lateral sclerosis; Huntington's disease; developmental pervasive disorders; deafness; deafness; diabetes; aging; side effects of drugs that inhibit mitochondrial function. The compound according to claim 4.

6. Treatment, prevention or suppression of neurodegenerative diseases, non-alcoholic fatty liver disease, tumors, cancer, scleroderma, hepatic hemosiderosis, hepatic copper overload, alopecia, human infertility, acute pancreatitis, fibromyalgia, mitochondrial disorders, or conditions associated with mitochondrial dysfunction or mitochondrial diseases; or Treatment or prevention of cancer, which is liver cancer, pancreatic cancer or biliary tract cancer; or Treatment or prevention of non-alcoholic fatty liver disease, which is non-alcoholic fatty liver disease, non-alcoholic steatohepatitis or cirrhosis; or Treatment or prevention of kidney disease, which is kidney cancer or renal insufficiency; or The compound according to any one of claims 1 to 5 for use in tumors.

7. As an antibacterial agent; or In cosmetics, or supplements, or nutraceuticals, i.e., as an active ingredient for anti-aging agents; as an anti-wrinkle skin care product; The compound according to any one of claims 1 to 6 for use in maintaining pluripotent cell cultures and as a supplement for cell cultures.

8. Use as a muscle protector or for muscle recovery after physical exercise; or The compound according to any one of claims 1 to 7 for use as a probe in imaging studies.

9. Comprising any one of the compounds of claims 1 to 8 and a pharmaceutically acceptable carrier, adjuvant, excipient, diluent, or a combination thereof, Wherein the pharmaceutically acceptable carrier is selected from the following list: saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, urea, or a combination thereof; Wherein the adjuvant is selected from the following list: water-in-oil emulsion adjuvants, aluminum adjuvants, TLR-4 ligands, saponins, and combinations thereof; The composition, wherein the excipient is selected from the following list: glucose, lactose, sucrose, glycerol monostearate, sodium chloride, glycerol, propylene, glycol, water, ethanol, or a combination thereof.

10. A composition for use in a method for the treatment or prevention of neurodegenerative diseases, non-alcoholic fatty liver disease, tumors, kidney diseases, scleroderma, hepatic hemosiderosis, hepatic copper overload, alopecia, human infertility, acute pancreatitis or fibromyalgia, wherein the daily dose of the composition is 20 mg / day or 10 mg / day, the composition according to claim 9.

11. A nanocarrier or liposome comprising the compound of claims 1 to 8 or the composition of claim 9 or 10.

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