Anti-aging composition comprising oxazole analogue as active ingredient

Abstract

One embodiment of the present invention provides an anti-aging composition comprising an oxazole analogue as an active ingredient. The composition can attenuate various phenotypes caused by aging. Specifically, a composition according to one embodiment of the present invention can reduce reactive oxygen species (ROS) and reduce dependence on glycolysis. In addition, telomere length can be increased, and mitochondrial function can be restored by increasing mitochondrial membrane potential.

Description

Anti-aging composition containing an oxazole analogue as an active ingredient The present invention relates to an anti-aging composition, and more particularly, to an anti-aging composition comprising an oxazole analogue as an active ingredient. Research on aging has continued from the past to the present, as aging is considered a disease-causing factor. Cellular senescence is defined as a state in which cell division ceases permanently, and this is considered a fundamental mechanism of aging and a cause of age-related diseases such as cancer, arteriosclerosis, and degenerative arthritis. Early biologists hypothesized that tissue aging was caused by the suppression of the repair mechanisms of surrounding cells by cells that no longer divide. These cells, now called senescent cells (SNC), not only do not participate in the repair of damaged tissue, but their secreted substances are also known to induce inflammatory cell migration, restructuring of the extracellular matrix, unwanted cell death, and fibrosis. These senescent cells naturally accumulate with aging, accumulating damage to cellular tissue. Current research technologies can selectively eliminate senescent cells, offering significant potential for developing methods to prevent and treat numerous age-related diseases. These treatments broadly include senolysis, immune-mediated senescent cell removal, and neutralization of the SASP. Permanent senescent cell removal not only leads to a sustained decrease in SASP but also reduces the risk of cancer development if senescent cells survive. However, prior to applying these technologies, several considerations must be made. First, the method of inducing senescent cells from which cells and the disease models to be applied must be considered. Furthermore, when applying these technologies to mouse experiments, stress, surgery, or the use of aged mice can induce senescent cells. The method of induction is also crucial. However, research on aging has yet to yield results, necessitating the development of novel substances or methods capable of suppressing aging. The technical problem to be achieved by the present invention is to provide an anti-aging composition containing an oxazole analogue as an active ingredient. The technical problems to be solved by the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below. In order to achieve the above technical task, one embodiment of the present invention provides an anti-aging composition comprising an oxazole analogue represented by the following [chemical formula 1] as an active ingredient. [Chemical Formula 1] In an embodiment of the present invention, the composition can reduce ROS (Reactive Oxygen Species). In an embodiment of the present invention, the composition can reduce the dependence on the process (Glycolysis). In an embodiment of the present invention, the composition can increase telomere length. In an embodiment of the present invention, the composition can reduce mitochondrial mass. In an embodiment of the present invention, the composition can increase mitochondrial membrane potential. In an embodiment of the present invention, the composition can reduce lipofusin in the cytoplasm. In an embodiment of the present invention, the composition can bind to ENO1 (alpha-enolase) and phosphorylate ENO1. In order to achieve the above technical task, another embodiment of the present invention provides a pharmaceutical composition for preventing, suppressing or treating symptoms of a mitochondrial-related disease, comprising an oxazole analogue represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] In an embodiment of the present invention, the mitochondrial-related disease is a disease related to mitochondrial disorder, such as myoclonic epilepsy; Myoclonic Epilepsy Associated with Ragged-Red Fibers (MERRF), Leber's Hereditary Optic Neuropathy (LHON), Neuropathy ataxia and retinitis pigmentosa (NARP), Mitochondrial Myopathy, Encephalopathy, Lactacidosis, Stroke (MELAS), Leigh syndrome, Leigh-like syndrome, Dominant Optic Atrophy (DOA), Kearns-Sayre Syndrome (KSS), Maternally Inherited Diabetes and Deafness (MIDD), Alpers-Huttenlocher syndrome, Ataxia Neuropathy spectrum, Chronic Progressive External Ophthalmoplegia (CPEO), Pearson syndrome, Mitochondrial Neuro-Gastro-Intestinal Encephalopathy (MNGIE), Sengers syndrome, 3-methylglutaconic aciduria, Sensorineural Deafness, Neuroradiological Findings of Encephalopathy and Leigh-Like Syndrome (MEGDEL), Myopathy, Mitochondrial Myopathy, Cardiomyopathy,and encephalomyopathy, SURF1 (COX deficiency Leigh syndrome due to complex IV surfeit protein deficiency). In an embodiment of the present invention, the mitochondrial-related disease is a disease related to mitochondrial dysfunction, and may be selected from the group consisting of Friedreich's Ataxia (FRDA), renal tubular acidosis, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, developmental pervasive disorder, hearing loss, deafness, and diabetes. The oxazole analogue according to an embodiment of the present invention can inhibit aging. The effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the detailed description of the present invention or the composition of the invention described in the claims. Figure 1 shows the results of in-house HTS at concentrations of 4 μM and 8 μM. Compared with 4 μM KB2764, 8 μM KB2764 showed a relatively more significant increase in the results. *P<0.1, **P<0.01, Student's t-test. Mean ± SD, n=10. Figure 2 is a drawing showing the synthesis process of KB2764 of the present invention. Figure 3 is a drawing showing biotinylation of KB2764 of the present invention. Figure 4 is NMR data of the compound synthesized in the present invention. (a) is the NMR data of compound 2 measured at 600 MHz, CDCl3. 1 H NMR spectrum. (b) is the 600 MHz, CDCl3, of compound 3. 1 This is the H NMR spectrum. Figure 5 is NMR data of the compound synthesized in the present invention. (a) is the NMR data of compound 4 (KB2764) measured at 600 MHz, CDCl3. 1 H NMR spectrum. (b) is the 600 MHz, CDCl3, of compound 5. 1 This is the H NMR spectrum. Figure 6 is NMR data of the compound synthesized in the present invention. (a) is the NMR data of compound 6 measured at 600 MHz, CDCl3. 1 H NMR spectrum. (b) is the 600 MHz, CDCl3, of compound 7. 1 This is the H NMR spectrum. Figure 7 is NMR data of compound 8 (Biotinylated KB2764) synthesized in the present invention, specifically, compound 8 measured at 600 MHz, MeOD. 1 This is the H NMR spectrum. Figure 8 is HRMS data of compound 4 (KB2764). Figure 9 is HRMS data of compound 8 (Biotinylated KB2764). Figure 10 is HPLC data of compound 4 (KB2764). Figure 11 is HPLC data of compound 8 (Biotinylated KB2764). Figure 12 is a drawing showing the potential ability of KB2764 of the present invention to inhibit aging, and specifically, shows the results of quantitative analysis of cell proliferation using a cell proliferation assay. **P< 0.01, Student's t-test. Mean ± SD, n= 10. Figure 13 is a drawing showing the potential ability of KB2764 of the present invention to inhibit aging, specifically, showing cell proliferation evaluation on day 12 by varying the concentration of KB2764 (0-32 μM). ns (not significant), **P< 0.01, Student's t-test. Mean ± SD, n= 10. Figure 14 is a diagram showing the potential ability of KB2764 of the present invention to inhibit aging, and specifically, shows cell viability evaluation to investigate the toxicity of KB2764 at concentrations of 8 μM and 50 μM. ns (not significant), **P< 0.01, two-way ANOVA followed by Bonferroni's post-test. Mean ± SD, n= 3. Figure 15 is a diagram illustrating the potential anti-aging ability of KB2764 of the present invention. Specifically, it shows the morphology of senescent fibroblasts treated with DMSO or KB2764 for 12 days. When DMSO was applied to senescent fibroblasts, they exhibited a broad and flat shape, whereas when KB2764 was applied to senescent fibroblasts, they exhibited a thin shape. Scale bar: 100 μm. Figure 16 illustrates the potential anti-aging ability of KB2764 of the present invention. Specifically, it shows the results of cell morphology analysis on agarose medium to determine whether colonies were formed by drug treatment. Senescent fibroblasts treated with DMSO and KB2764 were compared with HeLa cells. Scale bar: 100 μm. Figure 17 is a drawing showing that KB2764 of the present invention attenuates the aging phenotype, specifically, the results of flow cytometric analysis of reactive oxygen species (ROS) using MitoSO. **P< 0.01, Student t-test. Mean ± SD, n= 3. Figure 18 is a drawing showing that KB2764 of the present invention attenuates the aging phenotype, specifically, the results of flow cytometric analysis of mitochondrial mass using MitoTracker green. **P< 0.01, Student t-test. Mean ± SD, n= 3. Figure 19 is a drawing showing that KB2764 of the present invention attenuates the aging phenotype, specifically, the results of flow cytometric analysis of autophagy flux using Cyto-ID green are shown. **P< 0.01, Student t-test. Mean ± SD, n= 3. ~. Figure 20 is a drawing showing that KB2764 of the present invention attenuates the aging phenotype, specifically, the results of flow cytometric analysis of mitochondrial membrane potential (MMP) using JC-10. **P< 0.01, Student t-test. Mean ± SD, n= 3.~ Figure 21 is a drawing showing that KB2764 of the present invention attenuates the aging phenotype, specifically, the results of flow cytometric analysis of lipofuscin in an unstained state. **P< 0.01, Student t-test. Mean ± SD, n= 3. Figure 22 is a drawing showing that KB2764 of the present invention attenuates the senescence phenotype. Specifically, it shows the results of DNA tail length analysis of senescent fibroblasts treated with DMSO and KB2764 for 12 days. The comet assay was used for DNA tail length analysis. **P< 0.01, Student t-test. Mean ± SD, n= 10. Figure 23 is a drawing showing that KB2764 of the present invention attenuates the aging phenotype, specifically, comparing DNA telomere length using quantitative PCR. **P< 0.01, Student t-test. Mean ± SD, n= 4. Figure 24 is a drawing showing that KB2764 of the present invention regulates the metabolic process of senescent fibroblasts. Specifically, it shows the results of comparing the pH change during glycolysis of senescent fibroblasts treated with DMSO for 12 days (senescent group on the graph). The comparison of glycolysis was performed using the extracellular acidification rate (ECAR) assay. **P< 0.01, two-way ANOVA followed by Bonferroni's post-test. Mean ± SD, n= 3. ~. Figure 25 is a diagram showing that KB2764 of the present invention regulates the metabolic process of senescent fibroblasts. Specifically, it shows the results of comparing the pH changes during glycolysis of senescent fibroblasts treated with KB2764, AZD7545 (AZD), and AZD alone for 12 days. The comparison of glycolysis was performed using the extracellular acidification rate (ECAR) assay. **P<0.01, two-way ANOVA followed by Bonferroni's post-test. Mean ± SD, n=3. Figure 26 is a diagram showing that KB2764 of the present invention regulates the metabolic process of senescent fibroblasts. Specifically, it shows the basal phenotype (bottom left of each group) and the stressed phenotype (top right of each group) of senescent fibroblasts treated with DMSO for 12 days (senescent group on the graph). The basal phenotype and stressed phenotype connected by dotted lines indicate metabolic potential. Mean ± SD, n = 3 Figure 27 is a diagram showing that KB2764 of the present invention regulates the metabolic process of senescent fibroblasts. Specifically, it shows the basal phenotype (lower left of each group) and the stressed phenotype (upper right of each group) of senescent fibroblasts treated with AZD or AZD alone together with KB2764 for 12 days. The basal phenotype and the stressed phenotype connected by the dotted line indicate the metabolic potential. Mean ± SD, n = 3 Figure 28 is the chemical structural formula of KB2764 of the present invention. Figure 29 is the chemical structure of biotinylated KB2764. Figure 30 is a schematic diagram of immunoprecipitation using biotinylated KB2764. Figure 31 shows the -10logP:-10log (p-value)), Avg. Mass (kDa): protein molecular mass average for pyruvate kinase M (PKM) and alpha-enolase (ENO1) proteins selected from a total of 382 proteins reacting with biotinylated KB2764. Ion Mobility Tandem Mass Spectrometer Time-of-Flight (IM-MS TOF) technology was used to detect a total of 382 proteins reacting with biotinylated KB2764. Figure 32 is a schematic diagram of the glycolysis process and proteins at each step. Figure 33 is a diagram showing co-immunoprecipitation of biotinylated KB2764 with Myc-tagged control, Myc-tagged pyruvate kinase M, or Myc-tagged alpha-enolase. The primary antibodies used in the immunoblotting process were Myc-tagged antibody and β-actin antibody. Figure 34 is a drawing showing that maintenance of PKM and ENO1 expression is essential for the rejuvenation mediated by KB2764 of the present invention. Specifically, (a) shows immunoblots of lysates from senescent fibroblasts transfected with lentivirus for control shRNA (control shRNA, shCTRL) or pyruvate kinase M shRNA (shPKM). The primary antibodies used in the immunoblotting process are anti-PKM antibody and β-actin antibody. (b) shows immunoblots of lysates from senescent fibroblasts transfected with lentivirus for control shRNA (control shRNA, shCTRL) or alpha-enolase shRNA (shENO1). The primary antibodies used in the immunoblotting process were anti-ENO1 antibody and anti-β-actin antibody. Figure 35 is a diagram showing that maintenance of PKM and ENO1 expression is essential for KB2764-mediated rejuvenation of the present invention. Specifically, it shows a comparison of pH changes during glycolysis in senescent fibroblasts transfected with shCTRL, shPKM, and shENO1. Comparison of glycolysis was performed using the ECAR assay. **P<0.01, two-way ANOVA followed by Bonferroni's post-test. Mean ± SD, n=3. Figure 36 is a diagram showing that maintenance of PKM and ENO1 expression is essential for KB2764-mediated senescence rejuvenation of the present invention. Specifically, it shows the results of comparing reactive oxygen species (ROS) in senescent fibroblasts transfected with shCTRL, shPKM, and shENO1 when treated with KB2764 of the present invention and when not treated. **P< 0.01, Student's t-test. Mean ± SD, n= 3. Figure 37 is a drawing showing that maintenance of PKM and ENO1 expression is essential for the rejuvenation mediated by KB2764 of the present invention. Specifically, (a) shows the results of microscale thermophoresis (MST) analysis to quantitatively evaluate the binding affinity between KB2764 of the present invention and pyruvate kinase M (PKM). The dissociation constant (K) of PKM d) was 9.32 μM, which shows the affinity between KB2764 of the present invention and PKM. (b) shows the results of microscale thermophoresis (MST) analysis to quantitatively evaluate the affinity (binding affinity) between KB2764 of the present invention and alpha-enolase (ENO1). The dissociation constant (K) of ENO1 d ) was 94.7 nM, which shows the affinity between KB2764 of the present invention and ENO1. Figure 38 is a diagram showing that KB2764 of the present invention regulates ENO1 phosphorylation, specifically, the effect of KB2764 of the present invention on ENO1 phosphorylation in cells. Cells were transfected with pCMV-Myc-ENO1 (WT) and then treated with DMSO or KB2764. Co-immunoprecipitation was then performed using an antibody targeting the Myc tag. Treatment with KB2764 of the present invention increased ENO1 (WT) phosphorylation (dotted box area). Figure 39 is a diagram showing that KB2764 of the present invention regulates ENO1 phosphorylation. Specifically, the use of multiple sequence alignments revealed the presence of conserved motifs in ENO1 across various species (human, Sumatran orangutan, mouse, cow). The use of a kinase prediction tool enabled the prediction of potential phosphorylation motifs in ENO1 (http: / www.cbs.dtu.dk / services / NetPhos / ). The ENO1 protein is known to contain putative phosphorylation sites at Ser 254, Ser 272, and Ser 353. Figure 40 is a drawing showing that KB2764 of the present invention regulates ENO1 phosphorylation. Specifically, it shows the results of an experiment in which cells were transfected with pCMV-Myc-ENO1 (WT) or pCMV-Myc-ENO1 (mutant construct), and then inoculated with DMSO or KB2764. Each transfected cell was co-immunoprecipitated with antibodies against Myc-tag. ENO1 (WT) and ENO1 (mutant construct). Phosphorylation was detected with p-SQ / TQ antibodies. Treatment with KB2764 of the present invention did not increase ENO1 (S254A) phosphorylation (dotted box area). Figure 41 shows the in silico binding mode of KB2764 of the present invention in alpha enolase (PDB ID: 3B97). The dotted lines indicate hydrogen bonding and cation-π reactions, respectively. KB2764 of the present invention was predicted to hydrogen bond with Ser 254 using oxazole (2.11 Å) and ester (2.14 Å) moieties. In addition, the 3-methylphenyl ring underwent a cation-π stacking reaction with Arg 253 (4.79 Å). Figure 42 shows immunoblotting of cross-linked ENO1. Enolase-1 and HRP-conjugated β-actin were used as primary antibodies. Figure 43 is a diagram confirming the in vivo KB2764-mediated aging rejuvenation of the present invention. Survival analysis was performed by continuously treating synchronized Caenorhabditis elegans (C. elegans) with DMSO or KB2764 (100 μM). KB2764 of the present invention extended the lifespan of C. elegans by an average of 3 days. **P<0.01, Kaplan-Meier analysis., n=60. Figure 44 shows lipofuscin observed under a fluorescence microscope in synchronized C. elegans treated with DMSO or KB2764 (100 μM) of the present invention for 10 days. More lipofuscin was observed in aged C. elegans (arrowheads). Figure 45 shows body bends observed on days 0, 7, and 14 after treatment of synchronized C. elegans with DMSO or KB2764 (100 μM) of the present invention. Body bends were measured as a single reciprocating motion and were evaluated as the number of times per minute. ns (not significant), *P<0.05, Student's t-test. Mean ± SD, n=5. Figure 46 illustrates the mechanism of action of KB2764, a compound of the present invention that has an anti-aging effect. KB2764 targets ENO1 and regulates metabolic processes. This controlled metabolic process restored the secretory phenotype associated with aging and aided DNA repair. KB2764 of the present invention demonstrated anti-aging effects both in vitro and in vivo. Hereinafter, the present invention will be described with reference to the attached drawings. However, the present invention can be implemented in various different forms and is therefore not limited to the embodiments described herein. In the drawings, irrelevant parts have been omitted for clarity of description, and similar parts have been designated with similar reference numerals throughout the specification. Throughout the specification, when a part is said to be "connected (connected, contacted, or coupled)" to another part, this includes not only cases where it is "directly connected," but also cases where it is "indirectly connected" with another part in between. Furthermore, when a part is said to "include" a component, this does not exclude other components, but rather implies that it may include other components, unless otherwise specifically stated. The terminology used herein is merely used to describe specific embodiments and is not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this specification, it should be understood that the terms "comprises" or "has" indicate the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but do not exclude in advance the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof. According to one aspect of the present invention, the present invention provides an anti-aging composition comprising an oxazole analogue represented by the following [chemical formula 1] as an active ingredient. [Chemical Formula 1] The above oxazole analogue is ethyl 5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylate (4); KB2764). High-Throughput Screening (HTS) is widely used to identify target compounds from compound libraries. In the present invention, HTS technology was used to measure the cell proliferation capacity of the compound library. Cell counts were determined using a method based on DNA content. Senescent fibroblasts from the compound library were treated with 8 μM of each compound, and cell proliferation was measured on day 12 (Figure 12). KB2764, the compound that showed the greatest increase in the number of senescent fibroblasts, was selected as a candidate compound (red bar in Figure 12). Figure 1 provides a comprehensive description of the reaction conditions and reagents used in the synthesis of KB2764. KB2764, as confirmed through HTS, may have other unintended effects in addition to attenuating senescence. Therefore, we further investigated whether KB2764 induces other changes in addition to attenuating cellular senescence. The cell proliferation effect of KB2764 was again evaluated at various concentrations (0-32 μM) (Figure 13). A marked increase in cell proliferation was observed at 2 μM, and a significant proliferation effect was observed at concentrations of 4-32 μM (Figure 13). The observed concentration-dependent effect on senescent fibroblast proliferation indicates that it was influenced by KB2764 (Figure 13). Next, the toxicity of KB2764 was confirmed by measuring cell viability from 0 to 20 days. Senescent fibroblasts treated with 8 μM KB2764 showed consistent toxicity compared to the negative control. In contrast, cells treated with 50 μM KB2764 showed decreased viability (Figure 14). Treatment with an appropriate concentration of KB2764 was not toxic (Figure 14). Cell viability decreased at a high concentration of KB2764 (50 μM), while the effect was minimal at a low concentration (2 μM). Therefore, the concentration of KB2764 used in the present invention is preferably 8 μM (Figure 1). One notable characteristic of aging is an increase in cell surface area. In one embodiment of the present invention, the effect of KB2764 on the morphology of senescent fibroblasts was investigated. After 12 days of treatment with 8 μM KB2764, senescent fibroblasts were observed using an inverted phase-contrast microscope. Senescent fibroblasts exhibited a broad, flat structure (Figure 15). In contrast, senescent fibroblasts treated with KB2764 exhibited small wrinkles, a characteristic feature of young cells (Figure 15). In one embodiment of the present invention, colony formation was confirmed, as drug-induced cell proliferation can induce tumorigenesis. HeLa cells, a well-known cervical cancer cell line, formed colonies after 12 days in a soft agar assay, whereas no colonies were observed in the aged cells and KB2764-treated group (Figure 16). These results indicate that KB2764 does not induce tumorigenesis. In summary, KB2764 was found to be non-toxic at intermediate concentrations and focused solely on attenuating aging. According to one embodiment of the present invention, the composition can reduce reactive oxygen species (ROS). Cellular aging caused by damaged mitochondria is characterized by the presence of excessive reactive oxygen species (ROS), which severely impairs cellular and organelle functions. Recent studies have confirmed that reducing ROS levels is an important target for attenuating aging (Tan, IL & Velarde, MC Targeting Mitochondria as a Strategy to Inhibit Cellular Senescence. Current Molecular Biology Reports 7, 20-29 (2021)). A significant reduction in cellular ROS can effectively explain the improvement of aging (Davalli, P., Mitic, T., Caporali, A., Lauriola, A. & D'Arca, D. ROS, Cell Senescence, and Novel Molecular Mechanisms in Aging and Age-Related Diseases. Oxid Med Cell Longev2016, 3565127 (2016)). In one embodiment of the present invention, ROS levels were measured in senescent fibroblasts treated with KB2764. Treatment with KB2764 significantly reduced ROS levels in senescent fibroblasts (Fig. 17). The reduction in ROS indicates the restoration of organelles, including mitochondria and the cell itself. According to one embodiment of the present invention, the composition can reduce dependence on glycolysis. Metabolic processes within cells generate adenosine triphosphate (ATP), an essential energy source for life. Aged cells are known to have difficulty generating ATP due to abnormal metabolic functions. The insufficient ATP is compensated for by excessive glycolysis. Therefore, in the present invention, the overall metabolic process of the cell was evaluated. Glycolysis was measured by the extracellular acidification rate (ECAR). The results showed that fibroblasts treated with KB2764 exhibited a lower dependence on glycolysis compared to senescent fibroblasts (Figure 24). It was confirmed that the decrease in ECAR caused by KB2764 was due to metabolic reprogramming. In the present invention, AZD7545 was applied as a pyruvate dehydrogenase kinase 2 inhibitor (PDK2), which effectively interferes with the acetyl-CoA cycle. Treatment with AZD7545 did not significantly change the ECAR ratio, and parallel treatment with KB2764 did not significantly change the ECAR ratio (Fig. 25). Based on the results showing that KB2764 may play a role in modulating this process, it was inferred that KB2764 may also play a role in regulating the efficiency of OXPHOS. To confirm this, oxygen consumption rate (OCR) was evaluated as an indicator of mitochondrial respiration. Senescent fibroblasts had slightly lower basal and maximal OCR than fibroblasts treated with KB2764, with no significant changes. However, AZD7545 treatment significantly reduced the OCR of fibroblasts, and KB2764 treatment significantly restored the decreased OCR values.Additionally, metabolic potential was assessed by comparing basal ECAR, OCR, and stress ECAR, OCR values. Fibroblasts treated with KB2764 were more active than senescent fibroblasts (Fig. 26). These data suggest that KB2764 increases the metabolic potential of treated fibroblasts. Senescent fibroblasts treated with AZD7545 exhibited a decrease in metabolic potential, while co-treatment with KB2764 restored this potential (Fig. 27). According to one embodiment of the present invention, the composition can increase telomere length. Telomeres are repetitive DNA sequences located at the terminal regions of chromosomes, which protect chromosomes from potential damage due to fraying and tangling. It is known that telomere length is shortened when DNA is damaged by external factors such as cell division or ROS. In one embodiment of the present invention, telomere length was measured using quantitative PCR. The telomere length of fibroblasts treated with KB2764 of the present invention was 24% longer than that of senescent fibroblasts (Figure 23). The significant increase in telomere length in fibroblasts treated with KB2764 indicates the repair of damaged DNA (Figure 23). According to one embodiment of the present invention, the composition can reduce the length of the nucleus. DNA plays a crucial role in life processes through replication and transcription. It is generally known that DNA is permanently damaged during cellular aging, accelerating aging. Damaged DNA exists in the nucleus in the form of small fragments. Therefore, after treating senescent fibroblasts with KB2764, a comet assay was performed to examine damaged DNA. Senescent fibroblasts exhibited significantly enlarged nuclei compared to fibroblasts treated with KB2764 (Figure 22). The decrease in nucleus length after KB2764 treatment indicated less DNA damage (Figure 22). According to one embodiment of the present invention, the composition can reduce mitochondrial mass. Increased mitochondrial mass and size are known to be a consequence of the aging process. Furthermore, the increase in mitochondrial mass is known to be caused by overexpression of ROS and a compensatory response to damaged mitochondria. In one embodiment of the present invention, mitochondrial mass was examined using Mito-Tracker green. Treatment with KB2764 significantly reduced mitochondrial mass, indicating that it inhibits the accumulation of dysfunctional mitochondria (Figure 18). According to one embodiment of the present invention, the composition can restore autophagy function. Autophagy, a crucial system for eliminating functions within cells, is for the decomposition of dysfunctional organs. Restoration of autophagy function is recognized as a key factor in attenuating aging. In the present invention, the status of autophagy function in aged cells was investigated. Autophagy flux was measured in cells treated with KB2764 using Cyto-ID green dye. Aged cells treated with KB2764 showed a significant increase in autophagy flux, indicating functional restoration of the autophagy system by KB2764 (Figure 19). In the present invention, indirect functional recovery of mitochondria was confirmed due to recovery of ROS, mitochondrial mass and autophagy function mediated by KB2764 of the present invention in aging fibroblasts. According to one embodiment of the present invention, the composition can increase mitochondrial membrane potential (MMP). Mitochondrial membrane potential (MMP) is known as an essential indicator of aging that maintains the electrochemical potential of hydrogen ions required for ATP synthesis through the electron transport chain. In one embodiment of the present invention, MMP was determined through JC-10 staining to confirm the recovery of mitochondrial function. It was confirmed that fibroblasts treated with KB2764 showed a significant increase in MMP compared to senescent fibroblasts (Figure 20). This result indicates direct mitochondrial recovery. In the present invention, indirect functional recovery of mitochondria was confirmed due to recovery of ROS, mitochondrial mass and autophagy function mediated by KB2764 of the present invention in aging fibroblasts. According to one embodiment of the present invention, the composition can reduce cytoplasmic lipofuscin. Lipofuscin is a yellow-brown fluorescent lipid-containing pigment that accumulates in the cytoplasm during cellular aging, and can be considered an indicator of cellular aging recovery. In one embodiment of the present invention, cytoplasmic lipofuscin was measured without staining. As a result, fibroblasts treated with KB2764 showed a significant decrease in cytoplasmic lipofuscin accumulation (Figure 21). KB2764 (Fig. 28) of the present invention was synthesized according to the method described in Fig. 2. Some IL-33 proteins associated with aging have not yet been clearly identified, and several studies have been conducted previously. In the present invention, proteins interacting with KB2764 were investigated to provide clues for the discovery of KB2764's interaction partner proteins and their functions. KB2764 biotinylated compounds (Figs. 3 and 29) were synthesized to obtain pull-down samples. Senescent fibroblasts were treated with 8 μM biotinylated KB2764 (biotinylated-KB2764) to obtain samples by streptavidin immunoprecipitation (streptavidin immunoprecipitation (IP)) (Fig. 30). IM-MS TOF experiments were performed to identify KB2764 and its interacting protein partners. 382 interacting proteins were identified in the sample. A detailed protein list is provided in Table 1 below. Candidate proteins were selected based on their affinity for KB2764 (as measured by p-value) and their association with aging. Summarizing the data, KB2764-induced metabolic changes were significant, with glycolysis-related proteins PKM (pyruvate kinase M) and alpha-enolase (ENO1) emerging as the most promising candidates (Figure 31). In particular, PKM and ENO1 are known to participate in glycolysis. PKM mediates the conversion of phosphophenol pyruvate to pyruvate, while ENO1 mediates the conversion of 2-phosphoglycerate to phosphophenol pyruvate (Figure 32). A pull-down assay was performed to confirm the interaction between the candidate proteins PKM and ENO1.The pCMV-Myc-PKM and pCMV-Myc-ENO1 plasmids were transfected into HEK293T cells, followed by treatment with biotinylated KB2764 and streptavidin-IP. The resulting cells were detected by immunoblotting using a Myc-tag antibody (Fig. 33). The immunoprecipitation (IP) results indicated that biotinylated KB2764 interacted with PKM and ENO1 proteins, confirming the IM-MS TOF results. Next, in silicodocking analysis was performed to predict which of the two candidates exhibited higher affinity. [Table 1] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] [Correction pursuant to Rule 91, October 17, 2024] The absence of a protein targeted by a compound provides the basis for identifying the target protein. In a previous study, the inventors of the present invention elucidated the compound-mediated senescence recovery mechanism by knocking down the target protein of the compound (Lee, YH et al. Targeting regulation of ATP synthase 5 alpha / beta dimerization alleviates senescence. Aging (Albany NY) 14, 678-707 (2022)). Therefore, to determine the roles of PKM and ENO1 in the senescence changes mediated by KB2764 of the present invention, the proteins were knocked down. After lentiviral infection of senescent fibroblasts with control shRNA (shCTRL), PKM shRNA (shPKM), and ENO1 shRNA (shENO1), protein knockdown was assessed and protein expression levels were evaluated. shRNA-mediated PKM and ENO1 knockdown decreased the endogenous expression levels of the proteins (Figures 34(a) and 34(b)). Furthermore, shRNA-mediated PKM and ENO1 knockdown significantly reduced the ECAR ratio in senescent fibroblasts (Fig. 35). These data indicate that PKM and ENO1 proteins can regulate the metabolism of senescent fibroblasts. Furthermore, the inventors of the present invention hypothesized that metabolic changes induced by the regulation of specific proteins through KB2764 might be associated with senescence. This hypothesis was confirmed by measuring senescence markers in PKM or ENO1 knockdown cells treated with KB2764. Control shRNA fibroblasts showed a significant decrease in ROS in response to KB2764 (Fig. 36(a)). Senescent fibroblasts with PKM knockdown showed a significant decrease in ROS upon KB2764 treatment (Fig. 36(a)). These results indicate that the effects of KB2764-induced senescence are independent of the presence or absence of PKM.However, senescent fibroblasts in which ENO1 was knocked down did not induce a decrease in ROS in response to KB2764 (Fig. 36(b)). These data demonstrate that ENO1 plays a crucial role in the KB2764-mediated anti-senescence effect. Subsequently, the affinity between KB2764 and ENO1 was determined by MST assay. His-tagged ENO1 was measured together with various concentrations of KB2764. The dissociation constant (K) was determined. d ) was measured as 94.7 nM, demonstrating a strong association between KB2764 and ENO1 (Fig. 37(a)). On the other hand, the dissociation constant between KB2764 and PKM was measured as 9.32 μM, indicating a small degree of interaction compared to ENO1 (Fig. 37(b)). In summary, the gene knock-down results suggest that KB2764 improves aging through the control of ENO1. The interaction between ENO1 and KB2764 has been confirmed in cells. However, the detailed mechanism of KB2764-mediated ENO1 regulation has not yet been investigated. Therefore, to investigate KB2764-mediated regulation of ENO1, we investigated the phosphorylation process. Identification of the novel phosphorylation site on ENO1 investigated here may help explain the glycolysis process in aging. Therefore, to investigate the phosphorylation process of ENO1, pCMV-Myc-ENO1 (WT) was transfected into HEK293T cells. Transfected HEK293T cells were treated with DMSO or KB2764, and then immunoprecipitated with a Myc-tag antibody. Immunoprecipitated ENO1 (WT) showed an increase in the phosphorylated form in response to KB2764 (box in Figure 38). The specific phosphorylation site of ENO1 mediated by KB2764 was investigated using the phosphorylation motif kinase prediction tool (http: / www.cbs.dtu.dk / services / NetPhos / ). In silico coding, KB2764 predicted a possible binding mode (biding mode) to form a hydrogen bond interaction with ENO1 (PDB ID: 3B97). The ENO1 Ser 254 residue was identified as a possible phosphorylation motif and showed minimal divergence between species (Fig. 39). To confirm the predicted phosphorylation site, the Ser 254 motif was point mutated to alanine (S254A). As controls, point mutated Ser 272 (S272A) and Ser 353 (S353A) were also performed (Fig. 39).Subsequently, pCMV-Myc-ENO1(WT), pCMV-Myc-ENO1(S254A), pCMV-Myc-ENO1(S272A), and pCMV-Myc-ENO1(S353A) were transfected, treated with DMSO or KB2764, and immunoprecipitated with Myc-tag antibody (Fig. 40). KB2764 treatment increased phosphorylation of ENO1 (WT, S272A, and S353A) (Fig. 40). In contrast, the predicted ENO1 (S254A) motif did not increase in response to KB2764, suggesting that KB2764-mediated phosphorylation of ENO1 occurs at Ser 254 (boxed area in Fig. 40). KB2764 predicted a possible binding mode to form a hydrogen bond interaction with ENO1 (PDB ID: 3B97). As shown in Figure 41, the oxazole and ester moieties interacted with Ser 254. Furthermore, the 3-methylphenyl moiety underwent a cation-π reaction with Arg 253 of ENO1 (Figure 41). These data suggest that KB2764 can tightly bind to ENO1 and is a potential target for its anti-senescence effect. Some proteins function by forming structural complexes within cells. ENO1 exists in monomer, dimer, and octamer forms within cells. ENO1 is functionally active in the octamer form. Therefore, immunoblotting was performed to confirm the KB2764-mediated conformational change of ENO1 after cross-linking. In KB2764-mediated senescent fibroblasts, the ENO1 octamer structure significantly increased compared to the negative control, while the ENO1 monomer structure significantly decreased compared to the negative control (Figure 42). These data suggest that the basic monomer unit assembles into the functional octamer complex.Therefore, according to one embodiment of the present invention, the composition can bind to ENO1 (alpha-enolase) and phosphorylate ENO1. In one embodiment of the present invention, aging changes due to KB2764 treatment were observed in fibroblasts. The present invention hypothesized that aging changes observed in vitro would also be observed in vivo. The use of C. elegans is suitable for studying aging regeneration due to its ease of observation and short life cycle. As with previous studies, a significant lifespan extension in C. elegans is significant. Therefore, KB2764-mediated lifespan extension in C. elegans was tested. For lifespan analysis, synchronized C. elegans were treated with DMSO or KB2764 (100 μM), and live populations were measured daily. The measured data were confirmed using Kaplan-Meier survival analysis. KB2764 treatment increased the median survival rate of C. elegans for 2 days, indicating a significant increase over the entire lifespan of approximately 20 days (Figure 43). Additional experiments were conducted to confirm the KB2764-mediated lifespan extension results. As in aging fibroblasts, lipofuscin accumulates in the cytosol during cellular senescence in C. elegans. A decrease in lipofuscin accumulation in C. elegans can also be considered an indicator of aging regeneration. Therefore, lipofuscin in C. elegans was examined using fluorescence microscopy. C. elegans treated with KB2764 accumulated less lipofuscin in their bodies compared to control C. elegans (Figure 44). The decrease in accumulated lipofuscin reaffirmed the KB2764-mediated lifespan extension in C. elegans. As aging progresses, activity in C. elegans decreases. For example, there are clear differences in body bending and movement between young and aged C. elegans. Therefore, C. elegans To quantify the activity of C. elegans, the body bending frequency per minute, body brightness, was measured.The luminance of synchronized C. elegans was assessed on days 0, 7, and 14 after exposure to DMSO or KB2764 (100 μM). On day 0, no statistically significant difference in luminance was observed between the two groups (Fig. 45). Conversely, a significant decrease in the activity of control C. elegans was observed over time on days 7 and 14 (Fig. 45). Although the activity of KB2764-treated C. elegans decreased significantly over time, it remained significantly higher than that of the control group (Fig. 45). The maintenance of high activity mediated by KB2764 supports the lifespan extension effect of C. elegans. Taken together, these results suggest that KB2764 treatment promotes recovery from aging in C. elegans. According to another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing, suppressing or treating symptoms of a mitochondrial-related disease, comprising an oxazole analogue represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] In the present invention, the condition associated with mitochondrial dysfunction or mitochondrial dysfunction may preferably be characterized by OXPHOS deficiency. All cells require energy. Therefore, energy deficiency affects the activity of all cells. In principle, all cells are affected by suboptimal amounts of one or more OXPHOS complexes. However, the actual suboptimal amount varies from cell to cell. For example, cells with relatively high energy expenditure, such as brain and muscle cells, typically require a higher amount of OXPHOS system than cells with low energy expenditure, such as residual T cells. Therefore, cells affected by OXPHOS deficiency are typically, but not limited to, muscle cells or brain cells. Mitochondrial disorders are pleiotropic in their clinical manifestations. Various tissues may be affected, such as the pancreas, heart, liver, eyes, inner ear, blood, colon, and kidneys. Additionally, cells from non-clinically affected tissues, such as fibroblasts, often exhibit mitochondrial defects. Cells affected by OXPHOS deficiency can have the OXPHOS deficiency inhibited, treated, or prevented by providing the KB2764 of the present invention to the cells. Specifically, a cell can be affected by OXPHOS deficiency if its OXPHOS capacity is lower than normal (i.e., in a comparable cell of the same type from a healthy individual). In addition to being derived from an individual with OXPHOS deficiency, several methods exist to determine whether a cell has an OXPHOS deficiency, including but not limited to, testing the oxygen consumption, ATP production capacity, and enzyme activity of individual OXPHOS complexes (Chretien and Rustin J Inherit Metab Dis. 2003;_26_(2-3): 189-98). According to one embodiment of the present invention, administration of KB2764 to cells can increase metabolic potential by modulating the OXPHOS process. Specifically, KB2764 of the present invention downregulates this process and increases mitochondrial membrane potential, thereby enabling the restoration of mitochondrial function and thereby enhancing metabolic potential. According to one embodiment of the present invention, the mitochondrial-related disease is a disease related to mitochondrial disorder, such as myoclonic epilepsy; Myoclonic Epilepsy Associated with Ragged-Red Fibers (MERRF), Leber's Hereditary Optic Neuropathy (LHON), Neuropathy ataxia and retinitis pigmentosa (NARP), Mitochondrial Myopathy, Encephalopathy, Lactacidosis, Stroke (MELAS), Leigh syndrome, Leigh-like syndrome, Dominant Optic Atrophy (DOA), Kearns-Sayre Syndrome (KSS), Maternally Inherited Diabetes and Deafness (MIDD), Alpers-Huttenlocher syndrome, Ataxia Neuropathy spectrum, Chronic Progressive External Ophthalmoplegia (CPEO), Pearson syndrome, Mitochondrial Neuro-Gastro-Intestinal Encephalopathy (MNGIE), Sengers syndrome, 3-methylglutaconic aciduria, Sensorineural Deafness, Neuroradiological Findings of Encephalopathy and Leigh-Like Syndrome (MEGDEL), Myopathy, Mitochondrial Myopathy, Cardiomyopathy,and encephalomyopathy, SURF1 (COX deficiency Leigh syndrome due to complex IV surfeit protein deficiency). According to one embodiment of the present invention, the mitochondrial-related disease is a disease related to mitochondrial dysfunction, and may be selected from the group consisting of Friedreich's Ataxia (FRDA), renal tubular acidosis, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, developmental pervasive disorder, hearing loss, deafness, and diabetes. In one embodiment of the present invention, the pharmaceutical composition may be prepared in any one of the following forms: injection, granule, powder, tablet, pill, capsule, suppository, gel, suspension, emulsion, drop or liquid, according to a conventional method. In one embodiment of the present invention, the pharmaceutical composition may further comprise one or more additives selected from the group consisting of suitable carriers, excipients, disintegrants, sweeteners, coating agents, bulking agents, lubricants, glidants, flavoring agents, antioxidants, buffers, bacteriostatic agents, diluents, dispersants, surfactants, binders, and lubricants commonly used in the manufacture of pharmaceutical compositions. Specifically, carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol,

[0031] Mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil can be used, and solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the above composition. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. According to one embodiment of the present invention, the pharmaceutical composition can be administered to a subject in a conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalation, topical, rectal, oral, intraocular or intradermal routes. The preferred dosage of the compound represented by the above chemical formula 1 may vary depending on the condition and weight of the subject, the type and severity of the disease, the drug form, the route and period of administration, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, although not limited thereto, the daily dosage may be 0.01 to 200 mg / kg, specifically 0.1 to 200 mg / kg, and more specifically 0.1 to 100 mg / kg. Administration may be administered once a day or divided into several doses, and the scope of the present invention is not limited thereby. In the present invention, the subject may be a mammal including a human, but is not limited to these examples. The present invention can provide articles of manufacture and kits containing substances useful for treating, preventing, or suppressing symptoms associated with conditions associated with mitochondrial disorders or mitochondrial dysfunction. The articles of manufacture include a container having a label. Suitable containers include, for example, bottles, vials, and test tubes. The container can be formed from various materials, such as glass or plastic. The container holds a composition having an active agent effective for treating, preventing, or suppressing symptoms associated with conditions associated with mitochondrial disorders or mitochondrial dysfunction. The active agent in the composition is one or more of the compounds of the present invention. The label on the container preferably indicates that the composition is used for treating, preventing, or suppressing symptoms associated with conditions associated with mitochondrial disorders or mitochondrial dysfunction, and may also indicate directions for in vivo or in vitro use, as described above. Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings. Example 1. Synthesis of KB2764 and biotinylated KB2764 (1) General matters All chemicals and solvents were purchased from Sigma-Aldrich, TCI, and Acros and used without further purification. The reaction was carried out on pre-coated silica gel plates (Silica gel 60F 254 ; Merek, Darmstadt, Germany) were monitored by thin-layer chromatography (TLC) and visualized by UV254 light and KMnO4 staining for detection. Column chromatography was performed with silica gel (Silica gel 60, 230-400 mesh ASTM; Merck, Darmstadt, Germany). Nuclear magnetic resonance (NMR) spectra were recorded at room temperature on a Bruker UltraShield 600 MHz Plus (=1H, 600 MHz; 13C, 150 MHz) spectrometer. All chemical shifts are reported in ppm from tetramethylsilane (δ= 0) and are measured relative to the solvent in which the sample was analyzed (CDCl3: 1 δ7.26 according to H NMR, 13 δ77.0 by C NMR; MeOH-d4: 1 δ3.31 according to H NMR, 13 (δ49.0 based on C NMR). 1H NMR shift values ​​are reported as chemical shift (δ integral), multiplicities (s = singlet, br = broad, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublets, td = triplet of doublets, qd = quartet of doublets), coupling constants (J in Hz), and assignments. High-resolution mass spectrometry (HRMS) was recorded on an Agilent 6530 Accurate Mass Q-TOF LC / MS spectrometer. (2) KB2764 synthesis Reagents and conditions: (i) t-BuONO, CuCl2, acetonitrile, 80 ℃ 6 h; (ii) m-CH3PhB(OH)2, Pd(PPh3)4, toluene, H2O, K2CO3, 90 ℃ 16 h; (iii) 3-bromotoluene, Pd(OAc)2, P(o-tol)3, Cs2CO3, toluene, 90 ℃ 16 h. Ethyl 2-chlorooxazole-4-carboxylate (2) Ethyl 2-aminooxazole-4-carboxylate (5.00 g, 32.0 mmol) was added in portions to a stirred solution of tert-butyl nitrite (5.70 mL, 48.0 mmol, 1.5 eq) and copper(II) chloride (6.50 g, 48.0 mmol, 1.5 eq) in acetonitrile (150 mL). The mixture was then heated at 80°C for 6 h. The solution was cooled and then partitioned between dichloromethane (170 mL), water (80 mL), and concentrated hydrochloric acid (8 mL). The aqueous layer was further extracted with dichloromethane, and the combined organics were washed with dried (MgSO4) and Evaporated. The residue was purified by column chromatography on silica gel (Hexane / Et2O = 11:1 to 10:1, v / v) to obtain ethyl 2-chlorooxazole-4-carboxylate (3.09 g, 49%). R f 0.43 (Hexane / Et2O = 3:1 v / v). 1 H NMR (600 MHz, CDCl3) δ 8.19 (s, 1H), 4.40 (q, J = 7.2 Hz, 2 H), 1.39 (t, J = 7.2 Hz, 3 H). Ethyl 2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylate (3) To a solution of 4-(trifluoromethyl)phenylboronic acid (1259 mg, 6.63 mmol, 1.5 eq), Pd(PPh3) (630 mg) and ethyl 2-chlorooxazole-4-carboxylate (776 mg, 4.42 mmol) were added together with K2CO3 in water (H2O) (2 mL) in toluene (20 mL). The solution was stirred at 80°C under argon for 16 h. The solution was cooled and then partitioned between ethyl acetate and water. The aqueous layer was further extracted with ethyl acetate (2 x 30 mL), and the combined organics were washed with dried (MgSO4) and evaporated. The residue was purified by column chromatography on silica gel (Hexane / Et2O = 9:1 to 6:1, v / v) to obtain ethyl 2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylate (1.01 g, 80%). R f 0.55 (Hexane / Et2O = 1:1 v / v). 1 H NMR (600 MHz, CDCl3): δ 8.35 (s, 1H), 8.25 (d, J = 7.8 Hz, 2H), 7.75 (d, J = 8.4 Hz, 2H), 4.47 (q, J = 7.2 Hz, 2H), 1.44 (t, J = 7.2 Hz, 3H) H). Ethyl 5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylate (4) Ethyl 2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylate (303 mg, 1.06 mmol), 3-bromotoluene (154 μL, 1.27 mmol, 1.2 eq), cesium carbonate (518 mg, 1.6 mmol, 1.5 eq), tri(o-tolyl)phosphine (65 mg, 0.21 mmol, 0.2 eq), and palladium acetate (47.6 mg, 0.21 mmol, 0.2 eq) were added and stirred at 90°C for 16 h under argon gas. The solution was cooled and then partitioned between ethyl acetate and water. The aqueous layer was then extracted with ethyl acetate (2 x 30 mL), and the combined organics were washed with dried (MgSO4) and evaporated. The residue was purified by column chromatography on silica gel (Hexane / EtOAc = 9:1 to 6:1, v / v) to obtain ethyl 5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylate (348 mg, 88%). R f 0.67 (Hexane / EtOAc = 2:1, v / v) ( 1H NMR (600 MHz, CDCl3) δ8.28 (d,J= 8.2 Hz, 2H), 7.90 (d,J= 7.9 Hz, 2H), 7.75 (d,J= 8.3 Hz, 2H), 7.40 (dd,J= 11.5, 4.2 Hz, 1H), 7.32 (d,J= 7.4 Hz, 1H), 4.46 (q,J= 7.1 Hz, 2H), 2.46 (s, 3H), 1.43 (t,J= 7.1 Hz, 3H). HRMSm / z: calcd for C 20 H 16 F3NO3[M+H] + 376.1116; found, 376.1158. (3) 비오틴화(biotinylated) KB2764 synthesis Conditions: (i)1 M NaOH, EtOH, rt, 6 h; (ii) N-(tert-butoxycarbonyl)-1,6-diaminohexane, HOBt, EDC-HCl, DMF, rt, 6 h; (iii) TFA, DCM, rt, 3 h; (iv) Biotin, HOBt, EDC-HCl, DMF, rt, 6 h. 5-(m-Tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylic acid (5) To a solution of ethyl 5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylate (330 mg, 0.88 mmol) in ethanol (20 mL) was added 1 M NaOH (2.64 mL, 2.64 mmol, 3 eq), and the mixture was stirred at room temperature for 6 h under argon gas. The solution was evaporated under reduced pressure. The residue was acidified with water and 3 N HCl to a pH = 1–2. The solid was filtered and washed with hexane to obtain 5-(m-Tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylic acid (275 mg, 91%). R f 0.4 (DCM / MeOH = 10:1, v / v) (275 mg 91%) 1 H NMR (600 MHz, CDCl3) δ8.26 (d,J= 8.2 Hz, 2H), 8.11 (d,J= 7.9 Hz, 1H), 8.07 (s, 1H), 7.79 (d,J= 8.2 Hz, 2H), 7.43 (t,J= 7.7 Hz, 1H), 7.34 (d,J= 7.5 Hz, 1H), 2.47 (s, 3H). tert-Butyl(6-(5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamido)hexyl)carbamate (6) To a solution of 5-(m-Tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxylic acid (100 mg, 0.29 mmol), N-(tert-butoxycarbonyl)-1,6-diaminohexane (78 μL, 0.35 mmol, 1.2 eq), HOBt (62 mg, 0.46 mmol, 1.6 eq), and EDC-HCl (88 mg, 0.46 mmol, 1.6 eq) in DMF (10 mL) were added, and the mixture was stirred at room temperature under argon gas for 6 h. The solution was evaporated under reduced pressure. The residue was partitioned between ethyl acetate and water. The aqueous layer was further extracted with ethyl acetate. The combined organic layers were dried over MgSO4 and concentrated. The residue was purified by flash chromatography ((0-20% EtOAc / Hexane, 25 g silica gel cartridge) to give tert-butyl(6-(5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamido)hexyl)carbamate (135 mg, 88%). R f 0.04 (DCM / MeOH = 10:1, v / v) 1H NMR (600 MHz, CDCl3) δ8.23 (d,J= 8.2 Hz, 2H), 8.20 - 8.16 (m, 2H), 7.77 (d,J= 8.3 Hz, 2H), 7.43 - 7.36 (m, 2H), 7.27 (s, 1H), 4.52 (s, 1H), 3.47 (dd,J= 13.5, 6.9 Hz, 2H), 3.12 (d,J= 6.2 Hz, 2H), 2.46 (s, 3H), 1.70 - 1.64 (m, 2H), 1.57 (s, 2H), 1.51 (dt,J= 14.3, 7.2 Hz, 2H), 1.44 (s, 9H), 1.41 - 1.36 (m, 2H). N-(6-Aminohexyl)-5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamide (7) To a solution of tert-butyl(6-(5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamido)hexyl)carbamate (122 mg, 0.23 mmol), trifluoroacetic acid (3 mL) was added in CH2Cl2 (9 mL). The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was evaporated under reduced pressure to obtain N-(6-aminohexyl)-5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamide (52 mg, 52%). R f 0.03 DCM / MeOH = 10:1, v / v) 1H NMR (600 MHz, CDCl3) δ8.19 (d,J= 8.0 Hz, 2H), 7.93 - 7.88 (m, 2H), 7.76 (d,J= 8.1 Hz, 2H), 7.65 (s, 1H), 7.37 (t,J= 7.6 Hz, 1H), 7.29 (d,J= 7.4 Hz, 1H), 3.47 (d,J= 5.6 Hz, 2H), 2.98 (s, 2H), 2.42 (s, 3H), 1.67 (s, 4H), 1.42 (d,J= 30.0 Hz, 4H). N-(6-(5-((3S,4S,6R)-2-Oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexyl)-5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamide (8) To a solution of N-(6-Aminohexyl)-5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamide (40 mg, 0.11 mmol), biotin (33 mg, 0.13 mmol, 1.2 eq), HOBt (24 mg, 0.17 mmol, 1.6 eq), and EDC-HCl (34 mg, 0.17 mmol, 1.6 eq) in DMF (10 mL) were added, and the mixture was stirred at room temperature under argon gas for 6 h. The solution was evaporated under reduced pressure. The residue was partitioned between ethyl acetate and water. The aqueous layer was further extracted with ethyl acetate. The combined organic layer was dried over MgSO4 and concentrated. The residue was purified by automatic flash chromatography (0-20% EtOAc / Hexane) to give N-(6-(5-((3S,4S,6R)-2-Oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)heptyl)-5-(m-tolyl)-2-(4-(trifluoromethyl)phenyl)oxazole-4-carboxamide (40 mg, 54%). R f 0.4 (DCM / MeOH = 10:1, v / v) 1H NMR (600 MHz, MeOD) δ8.37 (d,J= 8.2 Hz, 10H), 8.13 (s, 5H), 8.08 (d,J= 8.0 Hz, 5H), 7.89 (d,J= 8.3 Hz, 10H), 7.42 (t,J= 7.7 Hz, 5H), 7.33 (d,J= 7.5 Hz, 6H), 4.60 (s, 3H), 4.47 (dd,J= 7.8, 4.8 Hz, 5H), 4.29 (dd,J= 7.8, 4.5 Hz, 5H), 3.44 (t,J= 7.1 Hz, 11H), 3.20 (d,J= 5.9 Hz, 11H), 2.92 (dd,J= 12.7, 5.0 Hz, 5H), 2.70 (d,J= 12.7 Hz, 5H), 2.46 (s, 15H), 2.20 (t,J= 7.3 Hz, 10H), 1.77 - 1.47 (m, 50H), 1.43 (dd,J= 15.4, 8.0 Hz, 30H), 1.31 (s, 9H). HRMSm / z: calcd for C 34 H 40 F3N5O4S [M+H] + 672.2792; found, 672.2767. 실시예 2. 세포 배양 Human dichromatic fibroblasts (HDF; PCS-201-010; ATCC, Manassas, VA, USA), human embryonic kidney cells (HEK293T; CRL-11268; ATCC), and Hela cells (A1100001; Thermo Fisher Scientific, Waltham, MA, USA) were used in this study. Cells were cultured in Dulbecco's modified Eagle's medium containing 25 mM glucose, 10% fetal bovine serum (SH30919.03; Hyclone, Waltham, MA, USA), 100 U / ml penicillin, and 100 μg / ml streptomycin (SV30079.01; Hyclone). Cell culture was performed using the same method as previously described. The doubling time of the population of senescent fibroblasts was measured when it exceeded 14 days. Cell viability was assessed using a Cedex HiRes Analyzer (05650216001; Roche, Basel, Switzerland). Example 3. Compound library screening Aged HDFs were cultured in 96-well plates at a density of 2,000 cells per well. Library compounds were diluted to a final concentration of 8 μM and added to the wells every 4 days. Cell proliferation was measured 12 days after drug treatment. Cell proliferation was measured using a previously described method. Example 4. Flow cytometry (FACS) FACS analysis was performed on a BD LSR II (LSR II; BD Bioscience; San Jose, CA, USA) at a core facility for in vivo cell imaging. Experimental methods using FACS are described in detail elsewhere in the specification. Example 5. Measurement of ROS, mitochondrial mass, and lipofuscin in HDFs To measure the amount of mitochondrial ROS, cells were incubated in medium containing 5 μM MitoSOX (M36008; Life Technologies, Carlsbad, CA, USA) for 30 min at 37°C. To measure mitochondrial mass, cells were incubated in medium containing 50 nM MitoTracker Green (M7514; Life Technologies) for 30 min at 37°C. Cells were left unstained for lipofuscin measurements in HDFs. Cells were prepared for FACS analysis using the method used in a previous study. Example 6. Measurement of mitochondrial membrane potential (MMP) To measure mitochondrial membrane potential, cells were incubated with 0.6 μg / ml JC-10 (ab112134; abcam, Waltham, MA, USA) for 30 min at 37°C. After staining, cells were prepared for FACS analysis as previously described. Example 7. Measurement of autophagy flux Autophagy flux was measured using the CYTO-ID autophagy detection method as previously described. Briefly, cells were incubated in medium containing 30 μM chloroquine (CQ; 455240250; Thermo Fisher Scientific). After 24 h of incubation, cells were stained with CYTO-ID Green Detection Reagent (CYTO-ID; ENZ-51031-0050; Enzo Lifescience, Farmingdale, NY, USA) and 50 nM LysoTracker™ Deep Red (LTDR; L12492; Invitrogen, Waltham, MA, USA) for 30 min and prepared for FACS analysis. To measure background autofluorescence, cells were incubated in medium without the dye. Fluorescence from CYTO-ID was normalized to fluorescence from LTDR. Autophagy flux was calculated using the following formula: Δmean fluorescence intensity (ΔCYTO-ID = MFI CYTO-ID (+CQ) / MFI CYTO-ID (-CQ). Example 8. Comet assay 1×10 trypsinized HDF 5After preparing the cells / ml concentration, the cells were washed with PBS. 50 μl of cells were mixed with 500 μl of dissolved LMAgarose. 75 μl of the mixture was placed on each comet slide and incubated for 10 min at 4°C in a light-protected manner. The hardened comet slides were immersed in lysis solution and incubated for 1 h at 4°C. The comet slides were transferred from the lysis solution to 1× TBE buffer and incubated for 30 min at 4°C. The comet slides were then electrophoresed at 35 V for 45 min. The electrophoresed comet slides were incubated with 70% ethanol for 5 min at room temperature. The fixed comet slides were dried for 15 min at 37°C. SYBR Green staining solution (diluted 1:1000 with distilled water) at 50 μl per well was observed under a FITC microscope (484 nm / 521 nm). Example 9. Measurement of telomere length Telomere length was measured by genomic DNA quantitative PCR (qPCR) analysis, as previously described. Telomere qPCR was performed using the primers listed in Table 2 below. [Table 2] [Correction pursuant to Rule 91, October 17, 2024] Example 10. Hippocampal analysis A Seahorse XFe96 analyzer (Aglient Technology, Santa Clara, CA, USA) was used according to the manufacturer's protocol. Briefly, 1 × 10 4Cells were dispensed into each well of XFe96 cell culture plates (101085-004; Aglient Technolongy) and cultured in a 5% CO2 incubator at 37°C for 16 h. The medium was then replaced with Seahorse XF DMEM pH 7.4 (103680-100; Aglient Technolongy). Extracellular acidification rate (ECAR) was measured using the Seahorse XF Glycolysis Stress Test kit (103020-100; Aglient Technolongy). Oxygen consumption rate (OCR) was measured using the Seahorse XF Cell Mito Stress Test Kit (103015-100 Aglient Technolongy). ECAR was reported in mpH / min, and OCR was reported in pmoles / min. The sensor cartridge was used in the Seahorse XFe96 Extracellular Flux Assay Kit (102416-100; Agilent Technology). Seahorse assays were performed as previously described. Example 11. IM-MS / MS TOF analysis Senescent HDFs were treated with 8 μM biotinylated KB2764 for 12 days. Immunoprecipitation and data analysis were performed according to conventional methods. Example 12. Plasmid production Plasmid pCMV-Myc-ENO1 was constructed by inserting the cDNA encoding human alpha-enolase (GenBank Accession No. NM_001428.5) into the pCMV-Myc vector (631604; Clontech, Mountain View, CA, USA). Plasmid pCMV-Myc-PKM was constructed by inserting the cDNA encoding pyruvate kinase M (GenBank Accession No. NM_001206796.3) into the pCMV-Myc vector. The His tag-Myc-ENO1 plasmid was prepared by mutagenesis of pCMV-Myc-ENO1 using HiFi DNA Assembly Master Mix (E2621L; New England Biolabs, Ipswich, MA, USA). The pCMV-Myc-ENO1 (mutant construct) plasmid was prepared by mutagenesis. The pLKO.1-shENO1 and pLKO.1-shPKM plasmids were prepared by inserting the target gene into the pLKO.1 plasmid. The primers used for plasmid construction are listed in Table 3. [Table 3] [Correction pursuant to Rule 91, October 17, 2024] Example 13. Immunoprecipitation (IP) pCMV-Myc-ENO1, pCMV-Myc-PKM, pCMV-Myc-ENO1 (mutant construct), and His-tag-Myc-ENO1 plasmids were transfected into HEK293T cells (CRL-11268; ATCC). Transfected cells were selected with 1 μM puromycin (BML-GR312-0050; Enzo Lifescinece) for 14 days. The medium was changed every 2 days during the selection period. After selection, immunoprecipitation was performed as described previously. Primary antibodies used were anti-Myc tag antibody (2276s; 1:1,000 dilution; cell signaling), anti-p-SQ / TQ antibody (2851s; 1:1,000 dilution; cell signaling), HRP-conjugated β-actin (sc47778; 1:2,000 dilution; Santa Cruz Biotechnology, Dallas, TX, USA), and HRP-conjugated anti-rabbit IgG (sc-2004; 1:2,000 dilution; Santa Cruz Biotechnology). Example 14. In SilicoDocking Analysis Ligand Preparation and Optimization: KB2764 was generated as a 2D and 3D structure using ChemDrawUltra (version 12.0.2, CambridgeSoft) and Maestro (version 13.4.134, MMshare version 6.0.134, Schrodinger), respectively. The 3D structure of the ligand was generated using the LigPrep module in the Maestro builder panel by adding hydrogen atoms, removing salts, and ionizing at pH (7 ± 2). Protein Preparation: The ENO1 structure (PDB ID: 3B97) was obtained from the Protein Database (https: / / www.rcsb.org). The protein structure was prepared using the protein preparation workflow in the Maestro panel. During protein preparation, bond order was specified and hydrogen atoms were added. Water molecules were removed. The receptor grid box was generated using the "Receptor Grid Generation" module in Glide. Binding analysis of KB2764 with ENO1 was performed using the Standard Precision ligand docking protocol of the Maestro panel. Example 15. Lentiviral production and infection HEK 293T cells were transfected with 8 μg of shRNA plasmids (pLKO.1-ctrl, pLKO.1-ENO1, and pLKO.1-PKM), 4 μg of PAX2 plasmid, and 4 μg of VSV.G plasmid using Lipofectamine 2000 (11668019; Invitrogen). Viral supernatants were harvested 24 h after transfection. Polybrene (TR-1003-G; 8 μg / ml; Millipore, Burlington, Middlesex County, MA, USA) was added to the viral supernatants. Viral infections were performed on aged HDFs as described previously. Example 16. Microscale thermophoresis (MST) test 6×His-tagged ENO1 protein was purified using the Capturem™ His-Tagged Purification Maxiprep Kit (635713; Takara, Mountain View, CA, USA). 200 nM of 6×His-tagged ENO1 protein was labeled using the Monolith His-Tag Labeling Kit RED-tris-NTA 2nd Generation (MO-L018; Nano Temper Technologies, München, Germany), using 0.4 mM KB2764. Microscale thermophoresis (MST) experiments were performed as described previously. Example 17. Crosslinking of PKM and ENO1 Aged HDFs were treated with DMSO and KB2764, and then lysed using cell lysis solution (87787; Thermo Fisher Scientific) at 4°C for 30 min. The lysate was buffer-exchanged with DPBS (21-031-CVC; Corning, NY, USA). Crosslinking was then performed with 5 mM bis(sulfosuccinimidyl) suberate (BS3; 21580; Thermo Fisher Scientific) at room temperature for 30 min. Crosslinked samples were boiled at 100°C for 10 min and then confirmed by immunoblotting. The primary antibodies used were enolase-1 (1:1,000; 3810T; Cell Signaling Technology), PKM2 (1:1,000; 4053T; Cell Signaling Technology), and HRP-conjugated β-actin (sc47778; 1:2,000 dilution; Santa Cruz Biotechnology). The secondary antibody used was HRP-conjugated anti-rabbit IgG (sc-2004; 1:2,000 dilution; Santa Cruz Biotechnology). Example 18. Reagents and media for C. elegans culture M9 buffer contained 5.8 g Na2HPO47H20, 3.0 g KH2PO4, 5.0 g NaCl, 0.25 g MgSO47H2O, and 1 L distilled water (DW). The M9 buffer was filtered through a 0.22 μm filter. A 20% alkaline hypochlorite solution was prepared by mixing 3.75 ml NaOH (1 M), 3.0 ml bleach, and 8.25 ml DW (distilled water). For nematode growth medium (NGM), 3 g NaCl, 2.5 g peptone, 17 g agar, and 975 ml DW (distilled water) were mixed and autoclaved. The bottle was cooled to 55°C, and 1 ml CaCl2 (1 M), 1 ml MgSO4 (1 M), 25 ml KPO4 (1 M, pH 6.0), and 1 ml cholesterol in ethanol (5 mg / ml) were mixed to form a plate. NGM medium cultured with OP50 (#053114; EDVOTEK, Washington, DC, USA) was prepared by inoculating OP50 into NGM medium and then culturing overnight in a 37°C incubator. Example 19. C. elegans culture and synchronization C. elegans (WT) were inoculated onto NGM medium cultured with OP50 and incubated at 22°C. After C. elegans were grown for 2–3 days, eggs and worms on the plates were prepared with 3 ml of M9 buffer. After sedimenting the eggs and worms for 10 min, the supernatant was removed. The plates were gently inverted for 5 min while completing with 15 ml of 20% alkaline hypochlorite solution. The plates were then centrifuged at 760 × g for 2 min and washed twice with M9 buffer. Finally, 7 ml of M9 buffer was added, gently shaken, and left overnight. The following day, L1-stage C. elegans were grown on NGM medium cultured with OP50. The NGM medium cultured with OP50 was replaced every 2 days. Example 20. C. elegans viability assay (C. elegans lifespan) Synchronized C. elegans were treated with 5-fluoro-2'-deoxyuridine (FUdR, 50 μM) to prevent egg laying. Synchronized L1-stage C. elegans were incubated at 22°C for 2–3 days. Adult C. elegans were transferred to NGM medium plates cultured with OP50 containing DMSO or KB2764 (100 μM). Viability assays were performed at 22°C, and the medium was changed every 2 days. The survival rate of C. elegans was observed microscopically and determined daily. Viability analysis was performed using the Kaplan-Meier method. Example 21. Measurement of lipofuscin in C. elegans Lipofuscin detection was performed on day 10 on NGM medium plates cultured with OP50 containing DMSO or KB2764 (100 μM). The presence of lipofuscin was observed using an X-Cite 120Q (Excelitas Technologies, Waltham, MA, USA) and a Zeiss AxioVert 200 (Zeiss, Oberkochen, Land Baden-Württemberg, Germany) instrument. Lipofuscin in C. elegans was observed at 450–490 nm. Example 22. Measurement of C. elegans body bends Luminance measurements were performed on day 10 on NGM medium plates cultured with OP50 containing DMSO or KB2764 (100 μM). The wriggling (based on reciprocation) of C. elegans was measured using a microscope for 1 min. Example 23. Statistical Analysis Statistical analysis was performed using a standard statistical software package (SigmaPlot 12.5; Systat Software, San Jose, CA, USA). Student's t-test and Bonferroni's posttest were used to determine whether differences were significant. Two-way ANOVA was performed for column-wise data analysis. The Kaplan-Meier method was validated in GraphPad Prism 7 software (GraphPad, San Diego, CA, USA). Significance between survival curves was assessed using the Kaplan-Meier method. The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single entity may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined manner. The scope of the present invention is indicated by the claims described below, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. An anti-aging composition comprising an oxazole analogue represented by the following [chemical formula 1] as an active ingredient. [Chemical Formula 1] 2. In paragraph 1, The composition is characterized in that it reduces ROS (Reactive Oxygen Species).

3. In paragraph 1, The composition is characterized in that it reduces the dependence on the process (Glycolysis).

4. In paragraph 1, The composition is characterized in that it increases telomere length.

5. In paragraph 1, The composition is characterized in that it reduces mitochondrial mass.

6. In paragraph 1, The composition is characterized in that it increases mitochondrial membrane potential.

7. In paragraph 1, The composition is characterized in that it reduces lipofusin in the cytoplasm.

8. In paragraph 1, The composition is characterized in that it binds to ENO1 (alpha-enolase) and phosphorylates ENO1.

9. A pharmaceutical composition for preventing, suppressing or treating symptoms of a mitochondrial-related disease, comprising an oxazole analogue represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] 10. In paragraph 9, The above mitochondrial-related diseases are diseases related to mitochondrial disorders, such as myoclonic epilepsy; Myoclonic Epilepsy Associated with Ragged-Red Fibers (MERRF), Leber's Hereditary Optic Neuropathy (LHON), Neuropathy ataxia and retinitis pigmentosa (NARP), Mitochondrial Myopathy, Encephalopathy, Lactacidosis, Stroke (MELAS), Leigh syndrome, Leigh-like syndrome, Dominant Optic Atrophy (DOA), Kearns-Sayre Syndrome (KSS), Maternally Inherited Diabetes and Deafness (MIDD), Alpers-Huttenlocher syndrome, Ataxia Neuropathy spectrum, Chronic Progressive External Ophthalmoplegia (CPEO), Pearson syndrome, Mitochondrial Neuro-Gastro-Intestinal Encephalopathy (MNGIE), Sengers syndrome, 3-methylglutaconic aciduria, Sensorineural Deafness, Neuroradiological Findings of Encephalopathy and Leigh-Like Syndrome (MEGDEL), Myopathy, Mitochondrial Myopathy, Cardiomyopathy, and EncephalomyopathyA pharmaceutical composition selected from the group consisting of SURF1 (COX deficiency Leigh syndrome due to complex IV surfeit protein deficiency).

11. In paragraph 9, The above mitochondrial-related disease is a disease related to mitochondrial dysfunction, and is a pharmaceutical composition selected from the group consisting of Friedreich's Ataxia (FRDA), renal tubular acidosis, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, developmental pervasive disorder, hearing loss, deafness, and diabetes.

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