KR20240076352A - Pharmaceutical composition for preventing or treating heart diseases related to sumoylation dysfunction comprising phenol compounds as active ingredients - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating heart disease related to SUMOylation dysfunction, which contains phenolic compounds as active ingredients, and among the phenolic compounds, salicylic acid, maltol, and esculetin. ) improves heart disease through activities such as increasing the contractility of myocardial cells and accelerating the rate of calcium breakdown within myocardial cells. In particular, the above-mentioned activity was found to be higher in myocardial cells with dysfunctional myocardial cells, and the above three types of phenolic compounds A composition for preventing, treating or improving heart disease related to hydrolysis dysfunction by confirming that a synergistic effect on the above activities appears when combined treatment; Alternatively, it can be usefully used as a composition for promoting sumoylation activity.

Description

Pharmaceutical composition for preventing or treating heart diseases related to sumoylation dysfunction comprising phenol compounds as active ingredients}

The present invention relates to a pharmaceutical composition for preventing or treating heart disease related to SUMOylation dysfunction, comprising a phenolic compound as an active ingredient.

Heart failure, a representative geriatric disease, has a variety of causes, and the prevalence and related medical burden are increasing. Traditionally, hypoglycemic agents are prescribed, and although the clinical symptoms of patients improve, the drugs that show long-term prognosis improvement are very limited. Therefore, the development of new mechanisms of treatment is urgently needed.

SUMOylation is an essential element for the normal function of cells and is a molecular mechanism that directly regulates the function of proteins, the basic unit of life. Abnormal sumoylation is known to be associated with the occurrence of various diseases. In patients with chronic heart failure, the function of sumoylation is reduced, and the deficiency of sumoylation in the heart reduces the enzyme activity and protein stability of the cardiomyocyte endoplasmic reticulum calcium pump (SERCA2a), which is a determinant of heart relaxation and contraction, causing heart failure and disease. It has been reported that it promotes the progression of Therefore, research on disease treatments through regulating sumoylation activity is actively underway.

1. Republic of Korea registered patent KR 10-1054943 (registered on August 1, 2011)

The purpose of the present invention is to prevent or treat heart disease related to SUMOylation dysfunction comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. To provide a pharmaceutical composition for use.

Another object of the present invention is to prevent or prevent heart disease related to SUMOylation dysfunction, comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. The purpose is to provide a health functional food composition for improvement.

Another object of the present invention is to provide a reagent composition for promoting SUMOylation activity containing at least one phenolic compound selected from the group consisting of salicylic acid, maltol, and esculetin as an active ingredient. It is provided.

In order to achieve the above object, the present invention relates to SUMOylation dysfunction comprising at least one phenolic compound selected from the group consisting of salicylic acid, maltol, and esculetin as an active ingredient. A pharmaceutical composition for preventing or treating heart disease is provided.

In addition, the present invention prevents or improves heart disease related to SUMOylation dysfunction, comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. Provides a health functional food composition for use.

In addition, the present invention provides a reagent composition for promoting SUMOylation activity containing as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. .

According to the present invention, among phenolic compounds, salicylic acid, maltol, and esculetin improve heart disease through activities such as increasing the contractility of myocardial cells and accelerating the rate of calcium breakdown within myocardial cells. In particular, the activity was found to be higher in cardiomyocytes with SUMOylation dysfunction, and when the three types of phenolic compounds were combined, a synergistic effect on the activity was confirmed to appear, thereby preventing heart disease related to SUMOylation dysfunction. Compositions for prevention, treatment or improvement; Alternatively, it can be usefully used as a composition for promoting sumoylation activity.

Figure 1 shows the results of analyzing the cytotoxicity of phenolic compounds in cardiomyocytes. ANOVA analysis shows statistically different values (*p<0.05). NT; no treatment, SyA; syringic acid, GA; gentisic acid, FA; ferulic acid and SA; salicylic acid.
Figure 2A shows the effect of phenolic compounds on ATP hydrolysis by SUMO activating enzyme E1 (SUMO E1); Figure 2B shows the effect of phenolic compounds on the cardiomyocyte endoplasmic reticulum calcium pump (hereinafter referred to as SERCA2a) SUMOylation; and Figure 2C shows the results of analyzing the effect of phenolic compounds on nuclear sumoylation. ANOVA analysis shows statistically different values (*p<0.05 and **p<0.005). NT; no treatment, SA; salicylic acid, CGA; chlorogenic acid and VA; vanillic acid.
Figures 3A to 3D show the results of analyzing changes in expression of BNP (Brain natriuretic peptide) and IL-6 (Interleukin-6) after treatment of phenol compounds in a heart failure cell model to confirm the effect of phenolic compounds on preventing or treating heart failure; Figures 3E and 3F show images of ISO-treated cardiomyocytes treated with a phenol compound and then observed under a microscope to confirm the effect of the phenolic compound on the change in the surface area of cardiomyocytes; and the results of analyzing changes in cell surface area. ANOVA analysis shows statistically different values (*p<0.05, **p<0.005, ***p<0.001 and ****p<0.0001). NT; no treatment, SA; salicylic acid, BNP; brain natriuretic peptide, IL-6; interleukin-6 and ISO; isoproterenol.
Figure 4 shows the results of comparing and analyzing the expression of genes related to sumoylation in heart tissue derived from normal mice (WT) and Duchenne muscular dystrophy (DMD) mice (hereinafter referred to as MDX mice) (n=3). T-test analysis shows statistically different values (*p<0.05 and **p<0.005).
Figure 5A shows the results of analyzing the calcium decay rate in cardiomyocytes derived from normal mice (WT) and MDX mice (MDX), and Figure 5B shows the results of analyzing the calcium decay rate after treating cardiomyocytes with phenolic compounds at different concentrations. . ANOVA analysis shows statistically different values (*p<0.05, **p<0.005 and ***p<0.001).
Figures 6A-6B show the contractile force of cardiomyocytes derived from MDX mice after treatment with phenol compounds alone or in combination; and the results of analyzing the calcium decay rate. ANOVA analysis shows statistically different values (*p<0.05 and **p<0.005). The combination treatment group (SA+M, M+E, and SA+M+E) was treated with 0.01 μM of phenol compound each. NT; no treatment, SA; salicylic acid, M; maltol and E; esculetin.

Hereinafter, the present invention will be described in more detail.

The present invention provides a pharmaceutical for preventing or treating heart disease related to SUMOylation dysfunction, comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. A composition is provided.

The phenolic compound can promote SUMOylation activity.

The promotion of SUMOylation activity may occur through promotion of the activity of sumoylation initiating enzyme (SUMO E1).

Additionally, the phenolic compound may inhibit the expression of Brain natriuretic peptide (BNP) or Interleukin-6 (IL-6), but is not limited thereto.

Additionally, the phenolic compound causes contraction of myocardial cells; Calcium breakdown within cardiomyocytes; and calcium release and re-sequestration from the myocardial endoplasmic reticulum into the cytoplasm.

The heart disease related to SUMOylation dysfunction is treated with isoproterenol (ISO); Alternatively, it may be caused by dystrophin deficiency.

The heart disease related to SUMOylation dysfunction may be one or more selected from the group consisting of Duchenne muscular dystrophy (DMD), heart failure, cardiac hypertrophy, cardiomyopathy, cardiac fibrosis, arrhythmia, and electrocardiogram disease, but is limited thereto. That is not the case.

Duchenne muscular dystrophy (DMD) is a hereditary progressive muscle disease caused by a deficiency of the dystrophin protein, causing problems in both skeletal and cardiac muscle, and abnormal intracellular calcium homeostasis in dystrophin-deficient muscles is a representative mechanism involved in disease progression. am. The MDX mouse model, which cannot express dystrophin protein due to a mutation in exon 23 of the dystrophin gene, is the most widely used preclinical model for Duchenne muscular dystrophy research.

The pharmaceutical composition of the present invention is prepared in unit dose form or in a multi-dose container by formulating it using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be manufactured by internalizing it.

The pharmaceutically acceptable carriers are those commonly used in preparation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, Includes, but is not limited to, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. In addition to the above ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc.

In the present invention, the content of additives included in the pharmaceutical composition is not particularly limited and can be appropriately adjusted within the content range used in conventional formulations.

The pharmaceutical compositions include injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, paste agents, and cataplasmase agents. It may be formulated in the form of one or more external skin preparations selected from the group consisting of, but is not limited to this.

The pharmaceutical composition of the present invention may additionally contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose, hydroxypropylcellulose, gelatin, alginate, polyvinyl pyrrolidone. It includes, but is not limited to, binders such as talc, calcium stearate, lubricants such as hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone and crospovidone, and surfactants such as polysorbate, cetyl alcohol, glycerol, etc. The pharmaceutically acceptable carrier and diluent may be biologically and physiologically friendly to the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents, and/or dispersion media.

The pharmaceutical composition of the present invention can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method. For oral administration, it can be formulated as tablets, troches, lozenges, aqueous suspensions, oily suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups, elixirs, etc. In the case of parenteral administration, it can be formulated as an injection, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.

The dosage of the pharmaceutical composition of the present invention is determined by the patient's condition, weight, age, gender, health, dietary constitution specificity, nature of the preparation, degree of disease, administration time of the composition, administration method, administration period or interval, excretion rate, and The range may vary depending on the drug form and can be appropriately selected by a person skilled in the art. For example, it may range from about 0.1 to 10,000 mg/kg, but is not limited and may be administered once or in divided doses several times a day.

The pharmaceutical composition may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally, or topically applied) depending on the desired method. The pharmaceutically effective amount and effective dosage of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time, administration route, etc. of the pharmaceutical composition, and those skilled in the art will know that it is effective for the desired treatment. Dosage can be easily determined and prescribed. The pharmaceutical composition of the present invention may be administered once a day, or may be administered in several divided doses.

In addition, the present invention prevents or improves heart disease related to SUMOylation dysfunction, comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. Provides a health functional food composition for use.

The present invention can be generally used with commonly used foods.

The food composition of the present invention can be used as a health functional food. The term “health functional food” refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with the Health Functional Food Act, and “functionality” refers to food that is related to the structure and function of the human body. It means ingestion for the purpose of controlling nutrients or obtaining useful health effects such as physiological effects.

The health functional food composition may contain common food additives, and its suitability as a “food additive” is determined in accordance with the general provisions and general test methods of the food additive code approved by the Ministry of Food and Drug Safety, unless otherwise specified. The decision is made based on the specifications and standards for the item.

Items listed in the “Food Additives Code” include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as subchromic pigment, licorice extract, crystalline cellulose, high-liquid pigment, and guar gum; Examples include mixed preparations such as sodium L-glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations.

The food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. For example, among health functional foods in the form of capsules, hard capsules can be manufactured by mixing and filling the composition according to the present invention with additives such as excipients in a regular hard capsule, and soft capsules can be manufactured by mixing and filling the composition according to the present invention. It can be manufactured by mixing with additives such as excipients and filling it with a capsule base such as gelatin. The soft capsule may contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., if necessary.

Definitions of terms such as excipients, binders, disintegrants, lubricants, coagulants, flavoring agents, etc. are described in literature known in the art and include those with the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in the conventional sense.

In the present invention, the term “prevention” refers to all actions that suppress or delay the above-mentioned heart disease by administering the composition according to the present invention.

In the present invention, the term “treatment” refers to any action that improves or beneficially changes the symptoms of heart disease by administering the composition according to the present invention.

In the present invention, the term “improvement” refers to any action that improves the bad condition of the heart disease by administering or ingesting the composition of the present invention to an individual.

In addition, the present invention provides a reagent composition for promoting SUMOylation activity containing as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. .

Hereinafter, the present invention will be described in detail through examples to aid understanding. However, the following examples only illustrate the content of the present invention and the scope of the present invention is not limited to the following examples. Examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

[ Preparation example 1] Experiment preparation

1-1. phenolic compounds

As an evaluation material, a low-molecular-weight single phenol compound with a purity of 98% or higher and a molecular weight of 500 or lower was tested for solubility in DMSO (dimethyl sulfoxide), and then a 10mM stock solution was obtained. The phenolic compounds are as shown in Table 1 below.

compound Molecular Weight Salicylic acid (SA) 138.12 Vanillic acid (VA) 168.2 Ferulic acid (FA) 194.19 Gentisic acid (GA) 154.1 Maltol 126.1 Syringic acid (SyA) 198.2 Esculetin (6,7-Dihydroxycoumarin) 178.14 Chlorogenic acid (CGA) 354.31

1-2. cell

H9c2 cells (ATCC, cat no.: CRL-1446), a rat cardiomyocyte cell line widely used in cardiovascular disease research, and AC16 cells (Sigma-Aldrich (cat no.: SCC109), a human left ventricle-derived myocardial cell line, and SUMOylation ) For the analysis, HEK-293 cells (ATCC, cat no.: CRL-1573), a human embryonic kidney-derived cell line, were used as primary adult cardiomyocytes (ACM) from 2-4 month old normal male mice. Jackson Laboratory, strain no.: 000476 C57/BL10ScSnJ) or Duchenne muscular dystrophy (DMD) mice (MDX mice) (Jackson Laboratory, strain no.: 001801 C57BL/10ScSn-Dmdmdx/J) were isolated. Afterwards, primary cardiomyocytes were separated from the digested heart tissue and resuspended in cardiomyocyte medium at low density, followed by retrograde perfusion with collagenase solution using a Langendorf device. and cultured under 5% CO ₂ conditions.

[ Experiment example 1] Cytotoxicity evaluation

To confirm the cytotoxicity of phenolic compounds in cardiomyocytes, H9c2 cells were treated with phenolic compounds at different concentrations (10 μM and 30 μM) for 24 hours, and cytotoxicity was measured using a WST (water soluble tetrazolium salt) kit (Dongin Biotech). Measured. Cytotoxic reactions were assessed as severe (<30%), moderate (30-60%), mild (60-90%), or non-cytotoxic (>90%) based on activity relative to the value obtained for the control. did.

[ Experiment example 2] Sumohwa active analysis

2-1. ATP hydrolysis analysis ( in vitro )

To determine the effect of phenolic compounds on SUMOylation, 10 μM of phenolic compounds were treated with SUMO activating enzyme E1 (purchased from R&D), and ATP hydrolysis was analyzed using the Promega kit. was carried out.

2-2. cardiomyocytes Endoplasmic reticulum calcium pump and nucleus Sumohwa analyze

To determine the effect of phenolic compounds on SUMOylation, HEK-293 cells overexpressing SUMO and SERCA2a (cardiomyocyte endoplasmic reticulum calcium pump), known as a substrate for sumoylation, were treated with 10 μM of phenolic compounds. , 24 hours later, the cells were lysed and immunoprecipitated with SUMO1 beads, and the SERCA2a protein bound to SUMO1 was separated using a 7.5% SDS-PAGE gel and Western blot was performed. Images were acquired using the LI-COR system. For nuclear sumoylation, HEK-293 cells overexpressing SUMO1 tagged with a fluorescent dye were treated with 10 μM of a phenol compound, and after 24 hours, the cells were imaged through a fluorescence microscope and the number of granules formed in the nucleus was measured using ImageXpress Pico. It was quantified using a system (Molecular Device).

[ Example 3] Analysis of cardiac function protection effect

3-1. Heart failure prevention effect analysis

To confirm the heart failure prevention effect of phenolic compounds, AC16 cells were cultured in 6-well plates with DMEM containing 10% FBS one day in advance, pretreated with phenolic compounds for 24 hours each, and treated with isoproterenol (hereinafter). After inducing heart failure through treatment (referred to as ISO), the expression levels of genes that promote cardiac cell hypertrophy and inflammatory response were measured through qRT-PCR. When treated with phenol compounds or isoproterenol, the medium was replaced with DMEM medium containing 1% FBS (DMEM with low FBS; hereinafter referred to as DMEMLF). The experimental group was classified as follows.

1) Untreated group (NT): Prepared by treatment with DMEMLF for 48 hours.

2) ISO only treatment group (ISO): Prepared by treatment with DMEMLF for 24 hours and 10 μM isoproterenol (hereinafter referred to as ISO) for 24 hours.

3) Phenol compound treatment group (ISO+SA, ISO+Maltol, and ISO+Esculetin): Prepared by treating 10 μM phenolic compound in DMEMLF for 24 hours, replacing it with DMEMLF medium containing 10 μM ISO, and culturing for 24 hours.

For qRT-PCR, RNA was extracted using TRIzol (Invitrogen), and cDNA was synthesized using the iScript cDNA synthesis kit (Bio-rad). qPCR was performed on a Qiagen real-time detection system using SYBR green PCR master mix (enzynomics), and changes in gene expression were compared, quantified by the ΔΔCt method and normalized to 18S ribosomal RNA.

3-2. Heart failure treatment effect analysis

To confirm the heart failure treatment effect of phenolic compounds, AC16 cells were cultured in 6-well plates with DMEMLF, treated simultaneously with ISO and each phenolic compound, and genes that promote cardiac cell hypertrophy and inflammatory response were identified through qRT-PCR. The expression level was measured. The experimental group was classified as follows. qRT-PCR was performed in the same manner as Experimental Example 3-1.

1) Untreated group (NT): Prepared by treatment with DMEMLF for 24 hours.

2) ISO only treatment group (ISO): Prepared by treatment with DMEMLF containing 10 μM ISO for 24 hours.

3) Phenol compound treatment group (ISO+SA, ISO+Maltol, and ISO+Esculetin): Prepared by treating DMEMLF containing 10μM ISO with 10μM phenol compound for 24 hours.

3-3. Cell surface area change analysis

To determine the effect of phenolic compounds on changes in the surface area of cardiomyocytes, ISO-treated AC16 cardiomyocytes were treated with 10 μM phenol compounds, and then cells were stained with alpha-actinin and observed under a microscope. Image J Cell surface area was quantified using software.

[ Experiment example 4] In cardiomyocytes Bioactive effects of phenolic compounds

To confirm the physiological activity effect of phenolic compounds on cardiomyocytes, single cardiomyocytes directly isolated from mouse hearts were treated with phenolic compounds using IonOptix imaging equipment (Milton, MA, USA), and then the sarcomere movement (contractile force) of the cells was measured. and changes in intracellular calcium levels were simultaneously tracked and analyzed in real time. Cardiomyocytes derived from normal mice and cardiomyocytes derived from MDX mice were used. Cardiomyocytes around 100 to 120 μm were identified to measure the degree of cell contraction, and intracellular calcium was loaded with Fura-2AM (0.5 μM, Invitrogen), deesterified, and measured through real-time imaging.

[ Experiment example 5] Statistical analysis

All experimental data were expressed as mean value ± standard error (mean + SEM). Statistical analysis used GraphPad Prism software, pairwise comparison used 2-tailed unpaired student's t-test, and differences between 3 or more groups were analyzed using 1-way or 2-way analysis of variance (ANOVA) using post-hoc test. was used. A P value of 0.05 or higher was considered statistically significant.

[ Example 1] Cytotoxicity analysis

According to Experimental Example 1, as a result of analyzing the cytotoxicity of phenolic compounds in cardiomyocytes, as shown in Figure 1, salicylic acid (SA), maltol, and esculetin were treated in two ways. It did not show cytotoxicity at any concentration. In addition, syringic acid (SyA), ferulic acid (FA), and gentisic acid (GA) each decreased or increased cell viability compared to the control group at a treatment concentration of 30 μM, but all were less than 10%. Therefore, it was judged to be non-cytotoxic (>90%).

[ Example 2] Sumohwa active analysis

2-1. ATP hydrolysis analysis ( in vitro )

According to Experimental Example 2-1, ATP hydrolysis analysis was performed to confirm the sumoylation activity of phenolic compounds. As shown in Figure 2A, salicylic acid (SA), maltol, and esculetin It was confirmed that it increases the ATP hydrolysis activity of sumoylation initiator enzyme (SUMO E1).

2-2. SERCA2a and nucleus Sumohwa analyze

According to Experimental Example 2-2, SERCA2a and nuclear sumylation analysis were performed to confirm the sumylation activity of phenolic compounds. As shown in Figures 2B and 2C, salicylic acid (SA), maltol, and Escul. It was confirmed that Esculetin increases SERCA2a and nuclear sumoylation. From the above results, it was confirmed that the three types of phenolic compounds have sumoylation activity ability in human cells.

[ Example 3] Analysis of cardiac function protection effect

3-1. Heart failure prevention effect analysis

According to Experimental Example 3-1, as a result of analyzing the heart failure prevention effect of the phenolic compound, as shown in Figures 3A and 3B, Brain natriuretic peptide (BNP) and Interleukin-6 (IL-6) in the ISO-only treatment group (ISO) 6) While expression increased, it was confirmed that the expression was suppressed in the three phenolic compound treatment groups (SA+ISO, Maltol+ISO, and Esculetin+ISO). From the above results, it was confirmed that the three types of phenolic compounds have a heart failure prevention effect.

3-2. Heart failure treatment effect analysis

According to Experimental Example 3-2, as a result of analyzing the heart failure treatment effect of the phenol compound, as shown in Figures 3C and 3D, Brain natriuretic peptide (BNP) and Interleukin-6 (IL-6) in the ISO only treatment group (ISO) 6) While expression increased, it was confirmed that the expression was suppressed in the three phenolic compound treatment groups (SA+ISO, Maltol+ISO, and Esculetin+ISO). From the above results, it was confirmed that the three types of phenolic compounds were effective in treating heart failure.

3-3. Cell surface area change analysis

According to Experimental Example 3-3, the effect of phenolic compounds on changes in cardiomyocyte surface area was analyzed. As shown in Figures 3E and 3F, the cell surface area increased in the ISO-only treatment group (IS0), whereas the three types Compared to the control group (NT), the group treated with phenolic compounds (salicylic acid, maltol, and esculetin) reduced the increased cell surface area caused by ISO, and it was confirmed that there was no significant difference between the phenolic compounds. From the above results, it was confirmed that the three types of phenolic compounds can inhibit cardiac hypertrophy caused by heart disease.

[ Example 4] In cardiomyocytes Analysis of physiological activity of phenolic compounds

4-1. MDX of mouse Sumohwa Dysfunction Analysis

In order to determine whether MDX mice have a dysfunction in medulosis, the expression of medulosis-related genes was analyzed in the heart tissue of MDX mice. As shown in Figure 4, it is involved in calcium metabolism in MDX mouse heart tissue (MDX), SUMO1 gene expression, a component of the sumoylation process, was significantly decreased compared to normal mouse heart tissue (WT), and expression of miR-146a, known as a regulatory mechanism that suppresses SUMO1 expression, was significantly decreased compared to normal mouse heart tissue (WT). increased significantly. From the above results, it was confirmed that MDX mice have sumoylation dysfunction.

4-2. MDX mouse origin In cardiomyocytes Analysis of physiological activity of phenolic compounds

According to Experimental Example 4, the physiological activity effect of phenolic compounds was analyzed in MDX mouse-derived cardiomyocytes. As shown in FIG. 5A, compared to normal mouse-derived cardiomyocytes (WT), MDX mouse-derived cardiomyocytes (MDX ), it was confirmed that the rate of calcium decay (tau) in the cytoplasm was slow.

In addition, as shown in Figure 5B, salicylic acid decreased the tau value by 3.47% compared to the control group (NT) in normal mouse-derived cardiomyocytes (WT) under the 0.1 μM treatment condition, whereas there was a significant change in the 1 μM treatment condition. There wasn't. On the other hand, in MDX mouse-derived cardiomyocytes (MDX), it was reduced by 23.13% and 27.70% under 0.1μM and 1μM treatment conditions, respectively. In particular, salicylic acid showed a tendency to increase the effect of improving tau values in a concentration-dependent manner.

In addition, maltol increased tau values by 11.87% and 9.37%, respectively, compared to the control group (NT) under 1 μM and 10 μM treatment conditions in normal mouse-derived cardiomyocytes (WT), but in MDX mouse-derived cardiomyocytes (MDX). In 1μM and 10μM treatment conditions, it was reduced by 27.91% and 29.85%, respectively, compared to the control group (NT).

In addition, Esculetin increased tau values by 20.48% and 25.24% in normal mouse-derived cardiomyocytes (WT) compared to the control group (NT) at 0.01 μM and 0.1 μM treatment conditions, respectively, but in MDX mouse-derived cardiomyocytes ( MDX) decreased by 23.78% and 14.40% compared to the control group (NT) at 0.01μM and 0.1μM treatment conditions. In particular, the tau value improved by salicylic acid, maltol, and esculetin treatment was similar to the tau value measured in the control group (NT) of normal mouse-derived cardiomyocytes (WT). From the above results, it was confirmed that the physiological activity of the three types of phenolic compounds increased more in myocardial cells with dysfunction than in normal myocardial cells.

[ Example 5] Synergy effect analysis through combined treatment of phenol compounds

In order to determine whether a synergistic effect on physiological activity in cardiomyocytes occurs when combined treatment of three types of phenolic compounds (salicylic acid, maltol, and esculetin), cardiomyocytes derived from MDX mice were treated with the three types of phenolic compounds alone or in combination. Afterwards, as a result of analyzing the physiological activity, as shown in Figure 6, the contractile force of cardiomyocytes was higher in the combination treatment group (SA+M, M+E, and SA+M+E) than the single treatment group even at a lower treatment concentration. It showed an excellent contractile effect, and it was confirmed that the combined treatment group showed a similar calcium decay rate even at a lower treatment concentration than the single treatment group. From the above results, it was confirmed that there is a synergistic effect through combined treatment of the three types of phenolic compounds in terms of physiological activity in cardiomyocytes.

As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. do. That is, the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (9)

A pharmaceutical composition for preventing or treating heart disease related to SUMOylation dysfunction, comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. The pharmaceutical composition according to claim 1, wherein the phenol compound promotes SUMOylation activity. The pharmaceutical composition according to claim 2, wherein the promotion of SUMOylation activity occurs through promotion of the activity of sumoylation initiating enzyme (SUMO E1). The pharmaceutical composition according to claim 1, wherein the phenol compound inhibits the expression of Brain natriuretic peptide (BNP) or Interleukin-6 (IL-6). The method according to claim 1, wherein the phenol compound promotes contraction of cardiomyocytes; Calcium breakdown within cardiomyocytes; And a pharmaceutical composition characterized in that it promotes one or more activities selected from the group consisting of calcium release and re-sequestration from the myocardial endoplasmic reticulum into the cytoplasm. The method according to claim 1, wherein the heart disease related to SUMOylation dysfunction is treated with isoproterenol (ISO); Or a pharmaceutical composition characterized in that it is caused by dystrophin deficiency. The method of claim 1, wherein the cardiac disease related to SUMOylation dysfunction is at least one selected from the group consisting of Duchenne muscular dystrophy (DMD), heart failure, cardiac hypertrophy, cardiomyopathy, cardiac fibrosis, arrhythmia, and electrocardiogram disease. Characterized pharmaceutical composition. A health functional food composition for preventing or improving heart disease related to SUMOylation dysfunction, comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin. . A reagent composition for promoting SUMOylation activity comprising as an active ingredient one or more phenolic compounds selected from the group consisting of salicylic acid, maltol, and esculetin.