Composition for ameliorating mitochondrial dysfunction comprising KS compound

Abstract

The present invention relates to a composition for ameliorating mitochondrial dysfunction, comprising a KS compound. In the present invention, it was confirmed that the KS compound ameliorates mitochondrial dysfunction by stabilizing manganese superoxide dismutase (MnSOD) protein or increasing SOD activity, and thus the KS compound of the present invention can be effectively used in a composition for preventing, ameliorating or treating mitochondrial dysfunction-associated diseases or diseases caused by oxidative stress.

Description

Composition for improving mitochondrial dysfunction containing KS compounds

[0001] The present invention relates to a composition for improving mitochondrial dysfunction comprising a KS compound.

[0002]

[0003] Mitochondria serve as the 'cell's power plant' that produces ATP, the cell's energy currency, and are also the site where reactive oxygen species (ROS), byproducts of oxidative phosphorylation, particularly superoxide anions (superoxide radicals), are most abundantly generated (Murphy, MP (2009). How mitochondria produce reactive oxygen species. Biochemical Journal, 417(1), 1-13). Under normal physiological conditions, mitochondria continuously generate superoxide radicals through electrons leaked from complexes I and III of the electron transport chain (ETC). Although ROS at appropriate concentrations function as intracellular signaling molecules, excessive accumulation leads to mitochondrial dysfunction by causing mutations in mitochondrial DNA (mtDNA), membrane lipid peroxidation, and protein oxidative damage (Wallace, DC et al., A mitochondrial paradigm of metabolic and degenerative diseases, aging, and cancer. Genetics, 171(4), 1397-1410, 2005).

[0004] To defend against oxidative damage to mitochondrial DNA caused by these reactive oxygen species, mitochondria possess a powerful antioxidant enzyme system, among which manganese superoxide dismutase (MnSOD or SOD2) plays a key role. MnSOD is the only antioxidant enzyme specifically present in the mitochondrial matrix and is responsible for the primary defense mechanism of converting harmful superoxide radicals into hydrogen peroxide and oxygen. A deficiency of MnSOD is known to cause severe mitochondrial damage, as well as fetal death, cardiomyopathy, and neurodegeneration, suggesting that MnSOD is a key factor in maintaining mitochondrial homeostasis (Holley, AK et al., Biology and Medicine, 50(10), 1253-1262, 2011).

[0005] According to recent studies, not only a simple quantitative decrease in MnSOD but also the failure to regulate the protein's structural stability and activity are identified as major causes of mitochondrial dysfunction in various pathological conditions. In particular, the activity of MnSOD is determined by its acetylation state, a post-translational modification (PTM) process; it has been revealed that SIRT3, a mitochondrial deacetylase, enhances enzyme stability and maximizes activity by deacetylating specific lysine residues of MnSOD (Tao, R. et al., Molecular Cell, 40(6), 893-904, 2010). In patient populations with aging, diabetes, cancer, and neurodegenerative diseases, the breakdown of this SIRT3-MnSOD axis leads to a decrease in MnSOD stability and, consequently, a loss of the ability to control oxidative stress (Holley, AK, et al. Manganese superoxide dismutase: guardian of the powerhouse. International Journal of Molecular Sciences, 12(10), 7114-7162, 2011).

[0006] Therefore, pharmacological approaches that enhance the structural stability of endogenous MnSOD proteins present within mitochondria or inhibit their degradation to maintain enzyme activity over a long period are regarded as innovative therapeutic strategies capable of fundamentally improving various metabolic and degenerative diseases associated with mitochondrial dysfunction. Unlike conventional antioxidants, which were limited to simply directly scavenging ROS and thus exhibited low efficiency, this invention aims to improve mitochondrial function through sustained antioxidant activity by restoring the structural integrity of the endogenous antioxidant enzyme MnSOD.

[0007]

[0008] In the present invention, as a result of diligent efforts to select drugs capable of improving mitochondrial dysfunction, it was confirmed that KS compounds bind to MnSOD (Manganese superoxide dismutase) to stabilize the tetramer structure of proteins, thereby removing reactive oxygen species and improving mitochondrial function through the restoration of enzyme activity, and the present invention was completed.

[0009]

[0010] Accordingly, the objective of the present invention is to provide a composition for the prevention, improvement, or treatment of diseases related to mitochondrial dysfunction comprising a KS compound as an active ingredient.

[0011] Another objective of the present invention is to provide an antioxidant composition comprising a KS compound as an active ingredient.

[0012]

[0013] In order to achieve the aforementioned purpose,

[0014] The present invention provides a pharmaceutical composition for the prevention or treatment of diseases related to mitochondrial dysfunction, or a health functional food composition for the prevention or improvement of diseases related to mitochondrial dysfunction, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0015] [Chemical Formula 1]

[0016]

[0017] (In the above chemical formula 1

[0018] R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and

[0019] R3 and R4 are -H or halogens, respectively.

[0020]

[0021] In a preferred embodiment of the present invention, the compound represented by Chemical Formula 1 may be a compound represented by the following Chemical Formula 1-1.

[0022] [Chemical Formula 1-1]

[0023] .

[0024] (In the above chemical formula 1-1, R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl.)

[0025] In another preferred embodiment of the present invention, the compound represented by Formula 1-1 is a compound represented by the following Formula 1-1A;

[0026] [Chemical Formula 1-1A]

[0027]

[0028] Compound represented by the following chemical formula 1-1B;

[0029] [Chemical Formula 1-1B]

[0030]

[0031] A compound represented by the following chemical formula 1-1C;

[0032] [Chemical Formula 1-1C]

[0033]

[0034] Compound represented by the following chemical formula 1-1D;

[0035] [Chemical Formula 1-1D]

[0036]

[0037] It may be one or more selected from the group consisting of.

[0038]

[0039] In another preferred embodiment of the present invention, the compound can stabilize the tetramer structure of MnSOD (Manganese superoxide dismutase).

[0040] In another preferred embodiment of the present invention, the compound can increase intracellular superoxide dismutase (SOD) activity and inhibit oxidative stress to restore the oxygen consumption rate (OCR) of mitochondria.

[0041] In another preferred embodiment of the present invention, the disease associated with mitochondrial dysfunction is any one or more ophthalmic diseases selected from the group consisting of age-related macular degeneration (AMD), glaucoma, cataract, diabetic retinopathy, proliferative non-retinopathy, retinitis pigmentosa, and dry eye syndrome; any one or more neurodegenerative diseases selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and dementia; any one or more muscular diseases selected from the group consisting of sarcopenia, proximal muscle weakness, and exercise intolerance; any one or more cardiovascular diseases selected from the group consisting of ischemic heart disease, heart failure, arteriosclerosis, and hypertension; any one or more cerebrovascular diseases selected from the group consisting of cerebral hemorrhage and cerebral infarction; and any one or more metabolic diseases selected from the group consisting of obesity, type 2 diabetes, metabolic syndrome, dyslipidemia, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis. One or more renal diseases selected from the group consisting of diabetic nephropathy and renal failure; one or more inflammatory and autoimmune diseases selected from the group consisting of chronic inflammation, inflammatory bowel disease, arthritis, rheumatoid arthritis, and Crohn's disease; one or more cancers showing mitochondrial metabolic abnormalities selected from the group consisting of glioblastoma, acute myeloid leukemia, renal cell carcinoma, adrenal ganglion, pheochromocytoma, breast cancer, colorectal cancer, melanoma, pancreatic cancer, and chronic lymphocytic leukemia; or one or more primary mitochondrial rare diseases selected from the group consisting of Leigh syndrome, MELAS syndrome, MURFF syndrome, and Leber hereditary optic neuropathy (LHON).

[0042]

[0043] In order to achieve other purposes,

[0044] The present invention provides an antioxidant composition comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0045] In a preferred embodiment of the present invention, the antioxidant composition may be a pharmaceutical composition for the prevention or treatment of oxidative stress-related diseases, or a health functional food composition for antioxidant purposes.

[0046]

[0047] To achieve another purpose,

[0048] The present invention provides a method for treating a disease associated with mitochondrial dysfunction or a disease caused by oxidative stress, comprising the step of administering a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0049] In a preferred embodiment of the present invention, the structural stability of MnSOD (SOD2) within the target cell can be enhanced by administering the compound.

[0050] In another preferred embodiment of the present invention, by administering the compound, the superoxide dismutase (SOD) activity within the target cell can be increased and oxidative stress suppressed, thereby restoring the oxygen consumption rate (OCR) of mitochondria.

[0051]

[0052] To achieve another purpose,

[0053] The present invention provides a topical skin composition for the prevention, improvement, or treatment of skin aging, comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0054] In a preferred embodiment of the present invention, the compound can improve skin aging by enhancing mitochondrial activity within skin cells.

[0055]

[0056] In the present invention, it has been confirmed that KS compounds improve mitochondrial dysfunction through the stabilization of MnSOD (Manganese superoxide dismutase) protein or an increase in SOD (superoxide dismutase) activity. Therefore, the KS compounds of the present invention can be usefully utilized as a composition for the prevention, improvement, or treatment of diseases related to mitochondrial dysfunction or diseases caused by oxidative stress.

[0057]

[0058] Figure 1 is data confirming the structural stabilization of MnSOD protein when aging RPTEC cells were treated with KS101 compounds at different concentrations.

[0059] Figure 2 shows data confirming MnSOD protein activity when aged RPTEC cells were treated with KS101 compound at different concentrations.

[0060] Figure 3 is data confirming the structural stabilization of MnSOD protein by the KS101 compound when purified MnSOD protein was heat-treated.

[0061] Figure 4 is data confirming the activity of MnSOD protein by the KS101 compound when purified MnSOD protein was heat-treated.

[0062] Figure 5 is data confirming whether the structure of the MnSOD protein was stabilized by four compounds having a structure similar to the KS compound when the purified MnSOD protein was heat-treated.

[0063] Figure 6 is a schematic diagram showing the process for confirming the binding of the KS101 compound and the MnSOD protein.

[0064] Figure 7 is data confirming that the KS101 compound specifically increases only SOD enzyme activity among various antioxidant enzymes.

[0065] Figure 8 is data confirming the reactive oxygen species inhibitory effect by KS101, KS102, KS103, and KS104 compounds.

[0066] Figure 9 is data confirming the oxygen consumption rate by KS101 compound, KS102 compound, KS103 compound, and KS104 compound.

[0067] Figure 10 is data confirming the effect of increasing aconitase activity by the KS101 compound.

[0068] Figure 11 is data confirming the α-ketoglutarate increasing effect by the KS101 compound.

[0069] Figure 12 shows data confirming the effect of the KS101 compound on increasing mitochondrial membrane potential (MMP) levels. Red indicates aggregated JC-1 fluorescent dye that cannot leak out of the mitochondria, and green indicates monomers that can leak out of the mitochondria.

[0070] Figure 13 shows data confirming that mitochondria enlarged due to aging are restored to normal levels by the KS101 compound: (A) Image of mitochondria (arrow) analyzed by transmission electron microscopy (TEM). Scale bar: 500 nm.; (B) Size of 20 mitochondria (μm²) measured using ImageJ software. 2 (c) The size distribution of 20 mitochondria was classified into various size ranges. The number of cells corresponding to each size category was recorded.

[0071]

[0072] The present invention will be described in detail below.

[0073]

[0074] In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of diseases associated with mitochondrial dysfunction, comprising as an active ingredient a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0075] In another aspect, the present invention relates to a health functional food composition for the prevention or improvement of diseases associated with mitochondrial dysfunction, comprising as an active ingredient a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0076] [Chemical Formula 1]

[0077]

[0078] (In the above chemical formula 1

[0079] R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and

[0080] R3 and R4 are -H or halogens, respectively.

[0081]

[0082] In the present invention, the compound represented by the above chemical formula 1 may be a compound represented by the following chemical formula 1-1.

[0083] [Chemical Formula 1-1]

[0084]

[0085] (In the above chemical formula 1-1, R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl.)

[0086] In the present invention, the compound represented by the formula 1-1 is a compound represented by the following formula 1-1A;

[0087] [Chemical Formula 1-1A]

[0088]

[0089] Compound represented by the following chemical formula 1-1B;

[0090] [Chemical Formula 1-1B]

[0091]

[0092] A compound represented by the following chemical formula 1-1C;

[0093] [Chemical Formula 1-1C]

[0094]

[0095] Compound represented by the following chemical formula 1-1D;

[0096] [Chemical Formula 1-1D]

[0097]

[0098] It may be one or more selected from the group consisting of.

[0099]

[0100] In the present invention, the compound can increase intracellular SOD activity and suppress oxidative stress to restore the oxygen consumption rate (OCR) of mitochondria.

[0101] In addition, the above compound can stabilize the tetramer structure of MnSOD (Manganese superoxide dismutase).

[0102] MnSOD (Manganese superoxide dismutase) is an antioxidant enzyme expressed from the SOD2 gene that exists within mitochondria and possesses antioxidant capacity. Oxidative stress occurs when the balance between the production of reactive oxygen species (ROS) and the cell's ability to neutralize their toxicity is disrupted. The types of reactive oxygen species include superoxide ions (O2 - ), hydrogen peroxide (H2O2), hydroxyl radical, ● OH), singlet oxygen, 1 A total of four types are known, including O2.

[0103] Among these, superoxide generated through the electron transport chain within mitochondria is neutralized by MnSOD to become H2O2, which then exits the mitochondria and is converted into water and oxygen by other antioxidant enzymes (peroxidase, catalase) abundant in the cytoplasm. However, because superoxide has low cell membrane permeability, it often fails to exit the mitochondria and remains inside; therefore, the role of MnSOD, the enzyme capable of removing it, is crucial. If it is not removed and remains, it damages the mitochondrial membrane, including mitochondrial DNA and proteins, leading to the initiation of cellular aging originating from the mitochondria.

[0104] In the present invention, we sought to determine whether a KS compound (Korean Registered Patent No. 10-2890732), known to induce Klotho expression, also affects MnSOD protein. As a result, it was confirmed that the KS compound specifically binds directly to MnSOD protein, stabilizing the tetramer structure of MnSOD protein and increasing MnSOD protein activity (Figs. 1 to 6). Furthermore, when the KS101 compound was reacted with various antioxidant enzymes, it was confirmed that the KS101 compound specifically increased only SOD enzyme activity (Fig. 7). Moreover, it was confirmed that the KS101, KS102, KS103, and KS104 compounds all inhibit reactive oxygen species (Fig. 8) and improve mitochondrial function, thereby increasing the oxygen consumption rate in senescent cells (Fig. 9).

[0105]

[0106] In the present invention, it has been confirmed that a KS compound directly binds to MnSOD in aged cells to protect against damage to the tetramer structure caused by aging or physical causes, thereby stabilizing it. Therefore, it can be used as a composition for the prevention, improvement, or treatment of diseases related to mitochondrial dysfunction caused by oxidative stress or aging. In addition, the MnSOD tetramer stabilized by binding with the KS compound can also be used as a composition for improving mitochondrial function.

[0107] In the present invention, mitochondrial dysfunction refers to a condition comprising at least one of the following: loss of mitochondrial membrane potential, reduced ATP synthesis, reduced MnSOD activity, or increased mtDNA mutations.

[0108] The aforementioned diseases associated with mitochondrial dysfunction include ophthalmic conditions such as age-related macular degeneration (AMD), glaucoma, cataracts, diabetic retinopathy, proliferative non-retinopathy, retinitis pigmentosa, and dry eye syndrome. The retina is one of the tissues in the human body with the highest mitochondrial density per unit area, and retinal pigment epithelial cells (RPEs) frequently suffer mitochondrial damage due to photo-oxidative stress caused by light and high energy requirements. MnSOD stabilizers inhibit oxidative damage to RPE cells, thereby preventing photoreceptor cell death, and can prevent glaucoma and other conditions by mitigating optic nerve mitochondrial damage caused by elevated intraocular pressure.

[0109] The diseases associated with the aforementioned mitochondrial dysfunction include neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, ALS, and dementia. O2 within mitochondria - Accumulation accelerates the aggregation of amyloid beta and induces neuronal death. MnSOD stabilization maintains the mitochondrial integrity of neurons, preventing synapse loss caused by oxidative stress, and can delay disease progression by suppressing neuroinflammatory responses.

[0110] Diseases associated with the aforementioned mitochondrial dysfunction include muscular system diseases such as senile sarcopenia, proximal muscle weakness, and exercise intolerance. Aged muscle cells experience reduced ATP production due to decreased mitochondrial function, and increased ROS activate the muscle protein degradation pathway (Ubiquitin-Proteasome system). MnSOD stabilization can improve the mitochondrial dynamics of muscle fiber cells, thereby preventing muscle mass loss and enhancing exercise tolerance.

[0111] Diseases associated with the aforementioned mitochondrial dysfunction include cardiovascular diseases such as ischemic heart disease, heart failure, arteriosclerosis, and hypertension, or cerebrovascular diseases such as cerebral hemorrhage and cerebral infarction. O2 generated in vascular endothelial cells - It reacts with nitric oxide (NO), a vasodilator, to produce harmful peroxynitrite (ONOO - It forms ) and reduces vascular elasticity. MnSOD is O2 - By preemptively removing NO, the bioavailability of NO is increased, and vascular endothelial function is protected, thereby protecting organs from arteriosclerosis and ischemic damage.

[0112] Diseases associated with the aforementioned mitochondrial dysfunction include metabolic diseases such as obesity, type 2 diabetes, metabolic syndrome, dyslipidemia, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis. Under conditions of overnutrition, the mitochondrial electron transport chain becomes overloaded, releasing large amounts of ROS. This can interfere with insulin receptor signaling (IRS-1) and induce insulin resistance. By stabilizing MnSOD to normalize ROS concentrations within mitochondria, the insulin signaling pathway can be restored, and the progression of fat accumulation and fibrosis in liver cells (NASH) can be blocked.

[0113] Diseases associated with the aforementioned mitochondrial dysfunction include kidney diseases such as diabetic nephropathy and renal failure. Under hyperglycemic conditions, the filtration function of kidney glomerular cells is destroyed and fibrosis progresses due to an increase in mitochondrial ROS. Maintaining the activity of MnSOD can prevent structural damage to glomerular podocytes, thereby reducing proteinuria and inhibiting the progression to chronic renal failure.

[0114] Diseases associated with the aforementioned mitochondrial dysfunction include chronic inflammation, inflammatory bowel disease, arthritis, rheumatoid arthritis, Crohn's disease, and other inflammatory and autoimmune diseases. ROS and mtDNA leaked from mitochondria activate the NLRP3 inflammasome, an intracellular inflammatory complex, thereby causing chronic inflammation. MnSOD stabilization can fundamentally regulate the secretion of excessive inflammatory cytokines (such as IL-1b) by blocking ROS, which is the upstream signal of NLRP3 activation.

[0115] The diseases associated with the aforementioned mitochondrial dysfunction may be cancers exhibiting mitochondrial dysfunction or metabolic abnormalities, and such cancers may include glioblastoma, acute myeloid leukemia, renal cell carcinoma, adrenal nodule, pheochromocytoma, breast cancer, colorectal cancer, melanoma, pancreatic cancer, chronic lymphocytic leukemia, etc. Cells prefer glycolysis over mitochondrial oxidative phosphorylation (OXPHOS) even when oxygen is sufficient. The composition of the present invention can act as a 'metabolic anticancer drug' that inhibits tumor growth by suppressing the abnormal energy metabolism of cancer cells through the normalization of mitochondrial function. Furthermore, mitochondria serve as the gateway to apoptosis. For cancer cells in which apoptosis signals are blocked due to mitochondrial dysfunction, the present invention can contribute to anticancer efficacy and overcoming anticancer drug resistance by restoring mitochondrial membrane potential through the stabilization and activation of MnSOD and initiating a normal apoptosis process. Furthermore, anticancer drugs such as cisplatin or doxorubicin cause potent mitochondrial toxicity, leading to cardiotoxicity, nephrotoxicity, and peripheral neuropathy. The composition of the present invention can be used as an adjuvant to alleviate the side effects of anticancer treatment by protecting MnSOD in normal cells.

[0116] Diseases associated with the above-mentioned mitochondrial dysfunction may include primary mitochondrial diseases (PMD). Primary mitochondrial diseases include diseases caused by mutations in mitochondrial DNA (mtDNA) or nuclear genes, such as MELAS syndrome, Leigh syndrome, MURFF syndrome, and Leber hereditary optic neuropathy (LHON). These rare diseases involve the explosive accumulation of superoxide radicals within mitochondria, accompanied by a severe ATP deficiency, which destroys high-energy organs such as the brain, muscles, and heart. The KS compound of the present invention can increase the survival rate and alleviate symptoms in patients with rare diseases by increasing overall SOD activity and raising the threshold for apoptosis.

[0117] In addition, it may include secondary mitochondrial damage resulting from exposure to drugs, toxins, smoking, alcohol, and environmental toxins; frailty syndromes associated with aging, such as general frailty, fatigue, reduced exercise tolerance, and weight loss; and acute and chronic mitochondrial dysfunction under metabolic stress. Although the above diseases exhibit different clinical manifestations, they share a common pathological mechanism of 'cell death and functional decline caused by increased mitochondrial ROS.'

[0118] In particular, the composition according to the present invention selectively stabilizes MnSOD protein within mitochondria of organs (heart, brain, retina, kidney, etc.) that have high energy metabolism requirements and generate large amounts of reactive oxygen species, thereby inhibiting apoptosis and fundamentally blocking the structural and functional degeneration of tissues; thus, an integrated therapeutic approach is possible for the broad group of mitochondria-related diseases listed above.

[0119]

[0120] In another aspect, the present invention relates to an antioxidant composition comprising a compound represented by the above formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0121] Since it has been confirmed that the KS compound of the present invention exhibits an antioxidant effect by increasing SOD (superoxide dismutase) activity, it can be utilized as a pharmaceutical composition for antioxidant purposes or a composition for a health functional food for antioxidant purposes.

[0122] The composition of the present invention is characterized by not only enhancing the stability of MnSOD, a mitochondrial-specific antioxidant enzyme, but also significantly increasing overall Total SOD activity within the cell. This effectively inhibits systemic cell damage and functional decline induced by oxidative stress by comprehensively removing reactive oxygen species generated in the cytoplasm and extracellular matrix as well as mitochondria-derived oxidative stress.

[0123] The above-described pharmaceutical composition for antioxidant purposes is a pharmaceutical composition for the prevention or treatment of diseases caused by oxidative stress. Such diseases caused by oxidative stress may include diseases related to the aforementioned mitochondrial dysfunction, as well as other skin diseases and photoaging such as wrinkles and loss of elasticity due to photoaging, pigmentation (melasma, age spots), atopic dermatitis, psoriasis, and vitiligo. Exposure to external ultraviolet (UV) rays and environmental pollutants causes a rapid increase in reactive oxygen species (ROS) within skin cells, which activates enzymes (MMP-1) that degrade collagen and damages elastic fibers. The composition of the present invention can prevent oxidative DNA damage and restore skin barrier function by normalizing SOD / MnSOD activity in the skin's dermis.

[0124] In addition, diseases caused by the aforementioned oxidative stress include chronic fatigue syndrome, fibromyalgia, systemic inflammatory response syndrome, and aging-related inflammation. Excessive ROS constantly activates the inflammatory signaling pathway (NF-κB), leading to the overproduction of cytokines and maintaining a systemic low-intensity inflammatory state. The potent antioxidant mechanism of the active ingredient can restore overall health by blocking the expression of inflammation-inducing factors at an upstream level.

[0125]

[0126] In another aspect, the present invention relates to a topical skin composition for the prevention, improvement, or treatment of skin aging, comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0127] In the present invention, the KS compound of the present invention can enhance mitochondrial activity within skin cells, so it is possible to prevent, improve, or treat skin aging.

[0128]

[0129] The pharmaceutical compositions of the present invention may be formulated into various forms according to conventional methods and used. For example, they may be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, and may be formulated into external preparations, suppositories, and sterile injectable solutions. Depending on each formulation, they may further include pharmaceutically acceptable carriers, excipients, and diluents. Additionally, they may be formulated into external preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, and sterile injectable solutions according to conventional methods and used.

[0130] The above carriers, excipients, and diluents include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. When the above pharmaceutical composition is formulated or prepared, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

[0131] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc., with the above composition. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Liquid dosage forms for oral administration include suspensions, liquid formulations, emulsions, syrups, etc., and may include various excipients, such as humectants, sweeteners, flavorings, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.

[0132] The pharmaceutical composition of the present invention may be prepared in a sterile injectable form and may be administered via various parenteral routes, such as intravenous injection, intramuscular injection, or subcutaneous injection. The injectable may be prepared according to conventional pharmaceutical techniques and consists of an active ingredient, a pharmaceutically acceptable carrier, and an additive. The pharmaceutically acceptable carrier used in the injectable form may include an aqueous solvent such as water for injection, physiological saline, glucose solution, Ringer's solution, or a mixture thereof. Optionally, a water-soluble co-solvent or a non-aqueous solvent, such as polyethylene glycol, propylene glycol, or glycerin, may be further included.

[0133] The injectable composition of the present invention may be adjusted to be isotonic using ionic or nonionic compounds, and examples include sodium chloride, glucose, mannitol, etc. Additionally, to ensure the stability and solubility of the composition, it may further include a buffer (e.g., phosphate buffer, citrate buffer), a pH adjuster (e.g., hydrochloric acid, sodium hydroxide), an antioxidant or chelating agent (e.g., sodium sulfite, EDTA), or a surfactant (e.g., polysorbate 80). For multi-dose formulations, preservatives within the conventional range, such as benzyl alcohol and parabens, may be included as needed. The injectable composition of the present invention is sterilized by sterile filtration, high-temperature sterilization, or an equivalent process, and then aseptically filled and sealed in a sterile ampoule or vial. The composition may be provided in the form of a concentrate diluted immediately before administration, an aqueous solution available for immediate administration, or a lyophilized powder; in the case of the lyophilized formulation, it is reconstituted by adding water for injection or an appropriate diluent before administration.

[0134] The term "administration" as used in the present invention means providing the pharmaceutical composition of the present invention to an individual by any appropriate method. The pharmaceutical composition of the present invention may be administered at a therapeutically effective dose, which is the amount of the active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal, or human as conceived by a researcher, veterinarian, physician, or other clinician, i.e., an amount that induces the alleviation of symptoms of the disease or disorder being treated. It is obvious to those skilled in the art that the therapeutically effective dose and frequency of administration of the pharmaceutical composition of the present invention will vary according to the desired effect. Therefore, the optimal dose to be administered can be easily determined by those skilled in the art and may be adjusted according to various factors including the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of formulation, the patient's age, body weight, general health condition, gender and diet, the time of administration, the route of administration and the secretion rate of the composition, the duration of treatment, and concurrently used drugs. The pharmaceutical composition of the present invention may be administered to an individual by various routes. For example, it may be administered intravenously, intraperitoneally, intramuscularly, intra-arterially, orally, intracardiaclysmically, intramedullaryly, transdermally, intestinally, subcutaneously, sublingually, or topically, but is not limited thereto. The pharmaceutical composition of the present invention may be administered in an amount of 1 to 10,000 mg / kg / day, and may be administered once a day or divided into several doses.

[0135] The composition of the present invention may be prepared in the form of a topical preparation applied directly to the skin and is provided in formulations such as creams, ointments, lotions, gels, essences, ampoules, patches, sprays, or masks. Such topical preparations are designed to promote skin penetration and absorption of active ingredients and include a pharmaceutically acceptable base that minimizes skin irritation. The base of the topical preparation may be composed of one or more of an oleaginous base, an aqueous base, a water-in-water (w / o) emulsion base, or an oil-water (o / w) emulsion base to effectively deliver active ingredients to the skin. The composition may include ethanol, isopropanol, propylene glycol, butylene glycol, glycerin, sorbitol, hyaluronic acid, or a mixture thereof as a solvent or humectant for skin penetration and moisturization. In addition, vegetable oils (jojoba oil, squalane, argan oil, etc.), fatty acids (linoleic acid, etc.), or ceramide derivatives may be added to moisturize the skin and strengthen the skin barrier. If necessary, viscosity enhancers (carboxymethylcellulose, hydroxyethylcellulose, carbomer, etc.), pH adjusters (citric acid, sodium hydroxide), antioxidants (butyrylhydroxytoluene, tocopherol), sunscreens, or skin emollients (allantoin, panthenol) may be included. As preservatives, parabens (methylparaben, propylparaben), phenoxyethanol, or natural extracts (grapefruit seed extract, etc.) are used in typical amounts.

[0136]

[0137] The health functional food composition of the present invention may be used as a health functional food, a food additive, or a dietary supplement. When the composition of the present invention is used as a food additive, it may be appropriately used according to conventional methods, such as by adding it as is or by mixing it with other foods or food ingredients.

[0138] In addition, the amount of the above-mentioned health functional food composition may be appropriately changed according to the purpose of use (prevention, health, or therapeutic treatment). As a specific example, when manufacturing food or beverages, the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, relative to the raw materials. However, when consumed for a long period for the purpose of health and hygiene or for health control, it may be added in an amount less than the above range, and since there are no issues regarding safety, the active ingredient may also be used in an amount greater than the above range.

[0139] There are no special restrictions on the types of food mentioned above, but examples of food to which the composition of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, etc., and include all health foods in the conventional sense.

[0140] When the health functional food composition of the present invention is manufactured into a beverage, it may include additional ingredients such as various flavoring agents or natural carbohydrates, as in conventional beverages. The natural carbohydrates may include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; natural sweeteners such as dextrin and cyclodextrin; and synthetic sweeteners such as saccharin and aspartame. The natural carbohydrates are included in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight, based on the total weight of the food composition of the present invention.

[0141] The health functional food composition of the present invention may include various nutritional agents, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., and may include fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages, but is not limited thereto. These ingredients may be used independently or in combination. Although the proportion of the above additives is not significantly limited, it is preferable that they be included in the range of 0.01 to 0.1% by weight relative to the total weight of the food composition of the present invention.

[0142]

[0143] The present invention will be explained in more detail below through examples.

[0144] These examples are solely for illustrating the invention, and it will be obvious to those skilled in the art that the scope of the invention is not to be interpreted as being limited by these examples.

[0145]

[0146] Example 1: Confirmation of MnSOD tetramer structural stabilization by KS101 compound treatment

[0147] Manganese superoxide dismutase (MnSOD) is composed of a tetramer structure (quaternary structure), and maintaining this structure is known to be crucial for the enzyme's activity in removing superoxide. In other words, if the protein's tetramer structure is disrupted, superoxide within the mitochondria is not removed and remains, which can accelerate mitochondrial aging.

[0148] There has been no detailed research on how to increase the enzyme activity of MnSOD. Accordingly, in the present invention, it was confirmed whether the compound KS101, which was identified as increasing TET enzyme activity, can also increase MnSOD enzyme activity.

[0149]

[0150] Relatively young (#4~6) RPTEC cells and aged (#11~12) RPTEC cells were treated with the compound KS101 at concentrations of 0 (DMSO), 0.1, 0.5, and 2.5 μM for 6 hours, and then the expression level of MnSOD protein was measured. Since MnSOD protein exists in monomeric to tetrameric structures, it is possible to confirm the structure based on protein size.

[0151] As a result, as shown in Figure 1, the amount of protein with a MnSOD tetramer structure increased due to treatment with the KS101 compound, and as shown in Figure 2, it was confirmed that MnSOD activity increased in a concentration-dependent manner with the KS101 compound.

[0152]

[0153] In addition, after reacting the purified MnSOD protein at 37°C and 50°C for 6 hours and confirming the protein structure through electrophoresis, as shown in Figure 3, unlike the MnSOD protein with DMSO added, no increase in monomer, dimer, or trimer forms of the protein was observed in the MnSOD protein with added KS101 compound even after incubation at 50°C.

[0154] Furthermore, when the enzyme activity was measured after reacting the purified MnSOD protein at 37°C and 50°C for 6 hours, as shown in Figure 4, the enzyme activity of the MnSOD protein with added KS101 compound did not decrease even after incubation at 50°C, whereas the enzyme activity of the MnSOD protein with added DM(DMSO) decreased to a statistically significant degree.

[0155] In other words, it was confirmed that the tetramer structure of the MnSOD protein is stabilized by the KS101 compound, so that the structure is maintained and the enzyme activity is maintained even under conditions where the structure of the MnSOD protein is damaged.

[0156]

[0157] Example 2: Confirmation of MnSOD tetramer structural stabilization by a compound having a structure similar to the KS compound

[0158] In order to confirm whether the MnSOD tetramer structure is stabilized by a compound having a structure similar to a KS compound, the experiment was performed in the same manner as in <Example 1> using KS compounds (KS101 compounds and KS104 compounds) and four compounds having a similar structure (purchased from Enamine).

[0159] Compounds having a structure similar to KS compounds are compound A (Z55890521, Enamine) represented by the following chemical formula A, compound B (Z55889103, Enamine) represented by the following chemical formula B, compound C (Z55890353, Enamine) represented by the following chemical formula C, and compound D (Z351264382, Enamine) represented by the following chemical formula D.

[0160]

[0161] [Chemical Formula A]

[0162]

[0163] [Chemical Formula B]

[0164]

[0165] [Chemical Formula C]

[0166]

[0167] [Chemical Formula D]

[0168]

[0169]

[0170] As a result, as shown in Figure 5, it was confirmed that the structural stability of the MnSOD tetramer was compromised under heating conditions of 50°C, but was maintained in the presence of KS101 or KS104 compounds. On the other hand, it was found that the MnSOD tetramer structure was not maintained in the case of other compounds with similar structures.

[0171] In other words, this means that the KS compound of the present invention selectively binds to a specific domain of the MnSOD protein, effectively stabilizing the tetrameric structure, which is the active form of the enzyme, even in a thermodynamically unstable state.

[0172]

[0173] Example 3: Confirmation of binding between KS101 compound and MnSOD protein

[0174] In the present invention, it was confirmed whether the KS101 compound binds to the MnSOD protein and stabilizes the MnSOD structure.

[0175] Compound KS101 was used to create a compound (compound 14 bait) linked to a biotin linker (control bait) to induce a reaction with aged renal tubular epithelial cell (RPTEC) extract. Subsequently, the biotin-bound compound was detached using streptavidin beads, and the proteins that detached along with it through this binding were identified and their nucleotide sequences analyzed (Fig. 6).

[0176] As a result, it was confirmed that among the proteins that do not appear in the control bait but are identified only in the KS101 compound (compound 14 bait), there is MnSOD protein (sequence in Fig. 6, sequence number 1).

[0177]

[0178] In addition, when examining how KS101 affects the activities of antioxidant enzymes SOD (superoxide dismutase), CAT (catalase), GPX (glutathione peroxidase), GR (glutathione reductase), and TrxR (thioredoxin reductase), it was confirmed that only the enzyme activity of SOD increased in a dependency on the concentration of the KS101 compound (Fig. 7).

[0179] In the present invention, it was confirmed that the reduction of reactive oxygen species and the increase of oxygen in cells by treatment with the KS101 compound depend on the enzymatic activity of SOD.

[0180]

[0181] Example 4: Confirmation of reactive oxygen species inhibitory efficacy by KS compound

[0182] In the present invention, the reactive oxygen species inhibitory effect of KS101 compound, KS102 compound, KS103 compound, and KS104 compound was confirmed.

[0183] First, RPTEC cells were seeded into 96-well white plates at a density of 100 µL to a density of 13,000 cells / well (repeated 4 times) and cultured overnight at 37°C under 5% CO2 conditions. The next day, the medium was discarded and washed with PBS; then, each compound was dissolved in 0.1% DMSO to a final concentration of 2.5 μM and 100 µL was added to each well (complete medium exchange). After incubating for 6 hours at 37°C under 5% CO2 conditions, the plates were washed with PBS, 100 µL of 10 μM ROS working solution was added to each well, and the plates were cultured for 30 minutes at 37°C under 5% CO2 conditions. Next, 10 µl / well of Hoechst 33342 (for confirming apoptosis) diluted 1:200 was added and incubated for 10 minutes (Hoechst staining was performed at a final 1:2000 by adding it to 100 µl / well of the existing medium). Afterward, all treatment reagents were discarded and washed with PBS, then 100 µl of PBS was added to each well, and ROS was measured (ROS=492nm / 527nm, Hoechst(DNA)=350 / 461nm). ROS concentrations were corrected for Hoechst staining and expressed as relative levels compared to the control group (DMSO treatment).

[0184] As a result, as shown in Figure 8, it was confirmed that compounds KS101, KS102, KS103, and KS104 all reduced the ROS concentration.

[0185]

[0186] Example 5: Confirmation of Increase in Oxygen Consumption Rate by KS Compound

[0187] In the present invention, it was confirmed how compounds KS101, KS102, KS103, and KS104 change the oxygen consumption rate (OCR) in the mitochondria of aged cells.

[0188] Aged renal tubular epithelial cells (#7 RPTEC) that had been subcultured seven times were treated with compounds KS101 to KS104 at a concentration of 1 μM each. The compounds were treated for two weeks up to subculture number 9 (#9), and 0.1% DMSO was treated in equal amounts as a control. In addition, young cells, renal tubular epithelial cells that had been subcultured four times (#4 RPTEC), were used as a comparison group.

[0189] Next, the oxygen consumption rate of each cell was measured using an OCR kit (Extracellular OCR Plate assay kit; Dojindo.E297). First, the cultured RPTEC cells were seeded into a 96-well black clear-bottom plate at a density of 30,000 cells / well using 100 µl of medium, and then incubated overnight at 37°C under 5% CO2 conditions. The next day, the medium was discarded, the plates were washed with PBS, and 90 µl of probe working solution was added to each well. After incubating for 30 minutes on a 37°C microplate reader, 10 µl of medium was added to each well, followed by incubation for 5 minutes on a 37°C microplate reader. Subsequently, fluorescence intensity was measured under the conditions Ex=500nm / Em=650nm. The measurement was performed using a bottom-reading method, reading from the bottom of the plate, and conducted in Kinetic mode to track real-time fluorescence changes at 10-minute intervals for a total of 120 minutes. Finally, the cell's oxygen consumption rate (OCR) was calculated by analyzing the slope of the measured fluorescence intensity over time.

[0190]

[0191] Fluorescence Intensity Measurement Values ​​50~80 mins #4 #9 #9+101 #9+102 #9+103 #9+104 OCR 6 1.1 1 17.0 3 6 1.9 3 2 3.3 3 4 5.9 8 5 5.6 2 (pmol / min) 6 7.9 5 2 6.3 2 3 5.3 1 2 8.2 1 5 0.6 3 4 8.8 4 0.1 4 2 4.7 9 2 4.1 3 2 6.7 8 6 0.0 4 4 3.3 3 6.2 3 3 6.3 7 5 3.8 5 2 5.6 7 3 8.0 1 4 1.5 9 2 0.8 8 2.5 7 56.1534.1628.07Avg45.2621.4243.832.0345.7643.48Ratio10.470.970.711.010.96Std0.420.280.380.30.230.23t-test 0.0242350.0290310.1182260.0050360.007983p-value *# ####

[0192] As a result, as shown in Figure 9, it was observed that the oxygen consumption rate (OCR) of mitochondria decreased significantly in aged cells subcultured 9 times compared to young cells subcultured 4 times. This implies that as cellular aging progresses, the basal respiratory activity and energy metabolism capacity of mitochondria functionally decline.

[0193] On the other hand, when aged cells were treated with the KS compounds of the present invention, it was confirmed that the OCR values, which had decreased in all treatment groups except KS102, including KS101, KS103, and KS104, were significantly increased. In particular, the compounds showed excellent efficacy in restoring the mitochondrial respiration function of aged cells to a level equivalent to that of young cells (P4).

[0194] In other words, this demonstrates that the KS compound of the present invention can reactivate the mitochondrial metabolic activity of renal epithelial cells whose function has declined due to aging. This suggests that the compound can serve as a powerful therapeutic agent capable of fundamentally improving various degenerative diseases and senility syndromes associated with mitochondrial dysfunction by going beyond simple oxidative stress control and reversing the energy production efficiency of aged cells to the level of young cells.

[0195]

[0196] Example 6: Measurement of aconitase activity

[0197] Aconitase activity was measured in RPTEC lysates prepared from cells treated with DMSO or KS101 compound for 6 hours. The protein concentration of the lysates was measured using the BCA assay. This analysis was performed using an aconitase activity test kit (ab109712, abcam) according to the manufacturer's instructions.

[0198] Aconitase activity was reduced in aged cells (#11) compared to young cells (#6), but it was confirmed that aconitase activity increased in a concentration-dependent manner after treatment with KS101. This suggests that mitochondrial function was improved by treatment with KS101 (Fig. 10).

[0199]

[0200] Example 7: α-ketoglutarate measurement

[0201] To measure α-ketoglutarate levels in RPTECs, cell lysates were prepared from cells treated with DMSO or KS101 compound for 6 hours. The protein concentration of the lysates was measured using the BCA assay and used to normalize the α-ketoglutarate measurements. The α-ketoglutarate concentration in RPTECs was measured using an α-ketoglutarate assay kit (MET-5131, Cell Biolabs) according to the manufacturer's instructions.

[0202] Although the amount of alpha-kedoglutate was reduced in aged cells (#14) compared to young cells (#5), it was confirmed that the amount of alpha-kedoglutate increased in a concentration-dependent manner after treatment with KS101. In other words, it can be seen that the amount of alpha-kedoglutate, a product of the tricarboxylic acid cycle (TCA cycle), increased through the improvement of mitochondrial activity (Fig. 11).

[0203]

[0204] Example 8: Measurement of Mitochondrial Membrane Potential (MMP)

[0205] Relative mitochondrial membrane potential (MMP) levels were measured in RPTECs treated with increasing concentrations of the KS101 compound. Young (#6) RPTECs were used as a control.

[0206] RPTECs were cultured in 96-well plates and treated with various concentrations of the KS101 compound for 6 hours under standard cell culture conditions. Then, JC-1 fluorescent dye (Mitochondria Staining Kit; CS0390, Sigma-Aldrich) was added to the cells and cultured for an additional 20 minutes. After washing the cells, the green fluorescence intensity of JC-1 monomers (assumed to be dispersed throughout the cell) was measured at an excitation wavelength of 490 nm and an emission wavelength of 530 nm. The red fluorescence intensity of JC-1 aggregates, representing dye aggregated in the mitochondrial matrix, was measured at an excitation wavelength of 525 nm and an emission wavelength of 590 nm. Changes in monomer and aggregate levels indicate changes in mitochondrial membrane potential (MMP). Relative MMP levels were expressed as the fluorescence intensity of monomers or aggregates as a percentage of the total fluorescence intensity.

[0207] It was confirmed that mitochondrial membrane potential (MMP) levels were decreased in aged cells (#12) compared to young cells (#6), but increased in a concentration-dependent manner after treatment with KS101 (Fig. 12). In Fig. 12, red represents fluorescent material remaining in the mitochondria, and green represents fluorescent material that has escaped. Higher red fluorescence indicates a higher mitochondrial membrane potential and thus higher function.

[0208]

[0209] Example 9: Measurement of Mitochondrial Size Levels

[0210] RPTEC cells were treated with 2.5 μM of compound KS101 or an equal amount of DMSO during subcultures #3–#8. RPTEC cells were harvested and fixed in glutaraldehyde at 4°C for 24 hours. After fixation, the samples were washed with 0.1 M phosphate buffer (0.1 M PB) and post-fixed in osmium tetroxide (OsO4) for 2 hours at room temperature (RT). Subsequently, dehydration was performed using stepwise ethanol solutions (50–100%). The dehydrated samples were embedded in epoxy resin and prepared into ultrathin sections approximately 70 nm thick using an ultrasectioning machine. The sections were stained with 6% uranyl acetate solution for 10 minutes and lead citrate solution for 5 minutes, followed by imaging. TEM analysis was performed using a JEM-1010 transmission electron microscope (JEM-1010, Japan) operating at an acceleration voltage of 80 kV. Images were taken at magnifications of 10,000x and 30,000x to visualize the morphology of mitochondria.

[0211] It was confirmed that the mitochondrial size level was increased in aged cells (#8) compared to young cells (#3), but decreased after treatment with KS101 (Fig. 13). This suggests that the mitochondrial size, which had become enlarged due to aging, was restored to a normal level.

[0212]

[0213] In the present invention, it has been confirmed that KS compounds improve mitochondrial dysfunction by stabilizing MnSOD (Manganese superoxide dismutase) protein or increasing SOD (superoxide dismutase) activity, so they can be usefully utilized as a composition for the prevention, improvement, or treatment of diseases related to mitochondrial dysfunction or diseases caused by oxidative stress.

Claims

1. A pharmaceutical composition for the prevention or treatment of diseases associated with mitochondrial dysfunction, comprising as an active ingredient a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] . (In the above chemical formula 1 R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and R3 and R4 are -H or halogens, respectively.

2. In Paragraph 1, A pharmaceutical composition for the prevention or treatment of diseases associated with mitochondrial dysfunction, characterized in that the compound represented by the above chemical formula 1 is a compound represented by the following chemical formula 1-1: [Chemical Formula 1-1] . (In the above chemical formula 1-1, R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl.) 3. In Paragraph 2, A pharmaceutical composition for the prevention or treatment of diseases associated with mitochondrial dysfunction, characterized in that the compound represented by the above chemical formula 1-1 is any one selected from the group consisting of the compound represented by the following chemical formula 1-1A; the compound represented by the following chemical formula 1-1B; the compound represented by the following chemical formula 1-1C; and the compound represented by the following chemical formula 1-1D: [Chemical Formula 1-1A] [Chemical Formula 1-1B] [Chemical Formula 1-1C] [Chemical Formula 1-1D] .

4. In Paragraph 1, A pharmaceutical composition for the prevention or treatment of diseases related to mitochondrial dysfunction, characterized in that the above compound stabilizes the tetramer structure of MnSOD (Manganese superoxide dismutase) to improve mitochondrial dysfunction.

5. In Paragraph 1, A pharmaceutical composition for the prevention or treatment of diseases associated with mitochondrial dysfunction, characterized by the above compound increasing intracellular SOD activity and suppressing oxidative stress to restore the oxygen consumption rate of mitochondria.

6. In Paragraph 1, The diseases associated with the above-mentioned mitochondrial dysfunction include: one or more ophthalmic diseases selected from the group consisting of age-related macular degeneration (AMD), glaucoma, cataract, diabetic retinopathy, proliferative non-retinopathy, retinitis pigmentosa, and dry eye syndrome; one or more neurodegenerative diseases selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and dementia; one or more muscular system diseases selected from the group consisting of sarcopenia, proximal muscle weakness, and exercise intolerance; one or more cardiovascular diseases selected from the group consisting of ischemic heart disease, heart failure, arteriosclerosis, and hypertension; one or more cerebrovascular diseases selected from the group consisting of cerebral hemorrhage and cerebral infarction; and one or more metabolic diseases selected from the group consisting of obesity, type 2 diabetes, metabolic syndrome, dyslipidemia, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis. A pharmaceutical composition for the prevention or treatment of diseases associated with mitochondrial dysfunction, characterized by being any one or more renal diseases selected from the group consisting of diabetic nephropathy and renal failure; any one or more inflammatory and autoimmune diseases selected from the group consisting of chronic inflammation, inflammatory bowel disease, arthritis, rheumatoid arthritis, and Crohn's disease; any one or more cancers exhibiting mitochondrial metabolic abnormalities selected from the group consisting of glioblastoma, acute myeloid leukemia, renal cell carcinoma, adrenal ganglion, pheochromocytoma, breast cancer, colorectal cancer, melanoma, pancreatic cancer, and chronic lymphocytic leukemia; or any one or more primary rare mitochondrial diseases selected from the group consisting of Leigh syndrome, MELAS syndrome, MURFF syndrome, and Leber hereditary optic neuropathy (LHON).

7. A health functional food composition for the prevention or improvement of diseases associated with mitochondrial dysfunction, comprising as an active ingredient a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] . (In the above chemical formula 1 R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and R3 and R4 are -H or halogens, respectively.

8. In Paragraph 7, A health functional food composition for the prevention or improvement of diseases related to mitochondrial dysfunction, characterized in that the compound represented by the above chemical formula 1 is a compound represented by the following chemical formula 1-1: [Chemical Formula 1-1] . (In the above chemical formula 1-1, R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl.) 9. In Paragraph 8, A health functional food composition for the prevention or improvement of diseases related to mitochondrial dysfunction, characterized in that the compound represented by the above chemical formula 1-1 is any one selected from the group consisting of the compound represented by the following chemical formula 1-1A; the compound represented by the following chemical formula 1-1B; the compound represented by the following chemical formula 1-1C; and the compound represented by the following chemical formula 1-1D: [Chemical Formula 1-1A] [Chemical Formula 1-1B] [Chemical Formula 1-1C] [Chemical Formula 1-1D] .

10. In Paragraph 7, A health functional food composition for the prevention or improvement of diseases related to mitochondrial dysfunction, characterized in that the above compound stabilizes the tetramer structure of MnSOD (Manganese superoxide dismutase) to improve mitochondrial dysfunction.

11. In Paragraph 7, A health functional food composition for the prevention or improvement of diseases related to mitochondrial dysfunction, characterized by the above compound increasing intracellular SOD activity and suppressing oxidative stress to restore the oxygen consumption rate of mitochondria.

12. In Paragraph 7, The diseases associated with the above-mentioned mitochondrial dysfunction include: one or more ophthalmic diseases selected from the group consisting of age-related macular degeneration (AMD), glaucoma, cataract, diabetic retinopathy, proliferative non-retinopathy, retinitis pigmentosa, and dry eye syndrome; one or more neurodegenerative diseases selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and dementia; one or more muscular system diseases selected from the group consisting of sarcopenia, proximal muscle weakness, and exercise intolerance; one or more cardiovascular diseases selected from the group consisting of ischemic heart disease, heart failure, arteriosclerosis, and hypertension; one or more cerebrovascular diseases selected from the group consisting of cerebral hemorrhage and cerebral infarction; and one or more metabolic diseases selected from the group consisting of obesity, type 2 diabetes, metabolic syndrome, dyslipidemia, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis. A health functional food composition for the prevention or improvement of diseases associated with mitochondrial dysfunction, characterized by being any one or more renal diseases selected from the group consisting of diabetic nephropathy and renal failure; any one or more inflammatory and autoimmune diseases selected from the group consisting of chronic inflammation, inflammatory bowel disease, arthritis, rheumatoid arthritis, and Crohn's disease; any one or more cancers showing mitochondrial metabolic abnormalities selected from the group consisting of glioblastoma, acute myeloid leukemia, renal cell carcinoma, adrenal ganglion, pheochromocytoma, breast cancer, colorectal cancer, melanoma, pancreatic cancer, and chronic lymphocytic leukemia; or any one or more primary rare mitochondrial diseases selected from the group consisting of Leigh syndrome, MELAS syndrome, MURFF syndrome, and Leber hereditary optic neuropathy (LHON).

13. A pharmaceutical composition for the prevention or treatment of diseases related to oxidative stress, comprising as an active ingredient a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] . (In the above chemical formula 1 R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and R3 and R4 are -H or halogens, respectively.

14. A health functional food composition for antioxidant purposes comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical Formula 1] . (In the above chemical formula 1 R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and R3 and R4 are -H or halogens, respectively.

15. A topical skin composition for the prevention, improvement, or treatment of skin aging, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical Formula 1] . (In the above chemical formula 1 R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and R3 and R4 are -H or halogens, respectively.

16. A method for preventing or treating a disease associated with mitochondrial dysfunction, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof to a subject requiring improvement or treatment of a disease associated with mitochondrial dysfunction: [Chemical Formula 1] . (In the above chemical formula 1 R1 and R2 are each -H, a halogen, or a C1-6 straight-chain or branched-chain alkyl, and R3 and R4 are -H or halogens, respectively.

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