Pharmaceutical composition for preventing or treating atherosclerosis

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating atherosclerosis and, more specifically, to a pharmaceutical composition for preventing or treating atherosclerosis that comprises a beta agonist or a pharmaceutically acceptable salt thereof, thereby promoting apoptotic cell removal and inhibiting plaque formation.

Description

Pharmaceutical composition for preventing or treating atherosclerosis

[0001] The present invention relates to a pharmaceutical composition for preventing or treating atherosclerosis.

[0002]

[0003] Atherosclerosis is a disease characterized by the development of lesions in the walls of blood vessels. These lesions can lead to narrowing of the artery walls due to the accumulation of atheromatous plaque. While atherosclerosis often has no symptoms in its early stages, as the disease progresses, it can lead to coronary artery disease, stroke, peripheral artery disease, or kidney failure, depending on the location of the affected artery.

[0004] Treatment for atherosclerosis involves dietary modification, eliminating risk factors such as smoking and alcohol abstinence, lipid-lowering drugs like statins, thrombolytics, and angioplasty. However, these treatments are dependent on the patient's will and are limited to temporary measures after the onset of the disease. Consequently, new treatment technologies and treatments are urgently needed.

[0005] Meanwhile, unlike normal tissues where apoptotic cells are rapidly removed, the removal of apoptotic cells is significantly reduced in lesioned blood vessels. This is due to a defect in efferocytosis, a cellular phenomenon in which apoptotic cells are removed by phagocytes such as macrophages. It has been revealed that this defect in efferocytosis leads to the accumulation of apoptotic cells, resulting in plaque formation and secondary necrosis of apoptotic cells within plaques, which in turn leads to the development of atherosclerosis.

[0006] The present inventors, taking note of the pathogenesis of atherosclerosis, conducted extensive research to treat atherosclerosis by promoting the removal of apoptotic cells and thereby inhibiting plaque formation, and completed the present invention.

[0007]

[0008] [Prior Art Literature]

[0009] [Patent Document]

[0010] (Patent Document 1) KR 10-2024-0070331 A

[0011] [Non-patent literature]

[0012] (Non-patent Document 1) Kojima Y, Weissman IL, Leeper NJ. The Role of Efferocytosis in Atherosclerosis. Circulation. 2017 Jan 31;135(5):476-489.

[0013] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating atherosclerosis.

[0014] The purpose of the present invention is to provide a health functional food for preventing or improving atherosclerosis.

[0015]

[0016] 1. A pharmaceutical composition for preventing or treating atherosclerosis, comprising a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof.

[0017] 2. In the above 1, the beta-agonist is any one selected from the group consisting of the following chemical formulas 1 to 6, a pharmaceutical composition for preventing or treating atherosclerosis:

[0018] [Chemical Formula 1]

[0019]

[0020] [Chemical Formula 2]

[0021]

[0022] [Chemical Formula 3]

[0023]

[0024] [Chemical Formula 4]

[0025]

[0026] [Chemical Formula 5]

[0027]

[0028] [Chemical Formula 6]

[0029]

[0030] 3. A pharmaceutical composition for preventing or treating atherosclerosis, wherein the prevention or treatment of atherosclerosis in the above 1 is achieved by promoting the removal of apoptotic cells.

[0031] 4. A pharmaceutical composition for preventing or treating atherosclerosis, wherein the prevention or treatment of atherosclerosis in the above 1 is achieved by inhibiting plaque formation.

[0032] 5. A pharmaceutical composition for preventing or treating atherosclerosis, further comprising a pharmaceutically acceptable carrier, excipient or diluent in the above 1.

[0033] 6. A health functional food for preventing or improving atherosclerosis, containing a beta-agonist or a food-based salt thereof.

[0034] 7. In the above 6, the beta-agonist is any one selected from the group consisting of the following chemical formulas 1 to 6, a health functional food:

[0035] [Chemical Formula 1]

[0036]

[0037] [Chemical Formula 2]

[0038]

[0039] [Chemical Formula 3]

[0040]

[0041] [Chemical Formula 4]

[0042]

[0043] [Chemical Formula 5]

[0044]

[0045] [Chemical Formula 6]

[0046]

[0047] 8. In the above 6, a health functional food that prevents or improves atherosclerosis by promoting the removal of apoptotic cells.

[0048] 9. A health functional food according to 6 above, wherein prevention or improvement of atherosclerosis is achieved by suppressing plaque formation.

[0049] 10. A method for preventing, treating, improving or suppressing atherosclerosis by administering a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof to an animal other than a human.

[0050]

[0051] The pharmaceutical composition and health functional food of the present invention are effective in promoting the removal of apoptotic cells by macrophages.

[0052] The pharmaceutical composition and health functional food of the present invention are effective in improving, inhibiting and / or treating atherosclerosis.

[0053] The pharmaceutical composition and health functional food of the present invention contain FDA-approved drugs with proven safety as active ingredients, and thus have fewer side effects, including adverse reactions.

[0054]

[0055] Figure 1 shows the results of a cytotoxicity test of ractopamine hydrochloride.

[0056] Figure 2 shows the results of an experiment to find the optimal concentration of ractopamine hydrochloride for promoting the removal of apoptotic cells.

[0057] Figure 3 shows the results of an experiment evaluating the efficacy of ractopamine hydrochloride in promoting the removal of apoptotic cells.

[0058] Figure 4 shows the results of an experiment evaluating the efficacy of ractopamine hydrochloride in promoting the removal of apoptotic cells in vivo.

[0059] Figures 5 to 7 show the results of experiments to confirm the mechanism of promoting the removal of apoptotic cells by ractopamine hydrochloride.

[0060] Figure 8 shows the experimental schedule for confirming the efficacy of ractopamine hydrochloride based on an animal model of atherosclerosis.

[0061] Figure 9 shows the results confirming the plaque size reduction effect of ractopamine hydrochloride in an animal model of atherosclerosis.

[0062] Figure 10 shows the results confirming the effect of ractopamine hydrochloride on reducing the size of necrotic cores in an animal model of atherosclerosis.

[0063] Figures 11 and 12 show the results of experiments confirming the efficacy of various beta-agonists in promoting the removal of apoptotic cells.

[0064]

[0065] The present invention provides a pharmaceutical composition for preventing or treating atherosclerosis, comprising a beta-agonist or a pharmaceutically acceptable salt thereof as an active ingredient.

[0066] In the present invention, the beta agonist is not limited to a specific type as long as it is a drug that acts on the beta adrenergic receptor. For example, beta-agonists include Denopamine, Dobutamine, Dopexamine, Ractopamine, Levosalbutamol, Ritodrine, Buphenine, Salmeterol, Salbutamol, Arformoterol, Orciprenaline, Isoetarine, Isoprenaline, Indacaterol, Xamoterol, Clenbuterol, Clorprenaline, Terbutaline, Fenoterol, Formoterol, Prenalterol, Procaterol, and Pirbuterol, and salts thereof.

[0067] In one embodiment, the beta-agonist can be ractopamine, salmeterol, indacaterol, cloprenaline, or procaterol, or a salt thereof.

[0068] In one embodiment, the beta-agonist can be ractopamine hydrochloride, salmeterol, salmeterol xinafoate, indacaterol, indacaterol maleate, cloprenaline hydrochloride, or procaterol hydrochloride.

[0069] In the present invention, “prevention” means any act of inhibiting or delaying atherosclerosis.

[0070] In the present invention, “treatment” means any action that improves or beneficially changes the symptoms of an individual suspected of having or having developed atherosclerosis.

[0071] In one embodiment, the pharmaceutical composition of the present invention can prevent or treat atherosclerosis by promoting apoptotic cell removal.

[0072] In one embodiment, the pharmaceutical composition of the present invention may prevent or treat atherosclerosis by reducing and / or inhibiting the formation of plaque.

[0073] In the present invention, “subject” means any animal, such as livestock and mice, that has developed or may develop atherosclerosis, and may be a mammal, including a human.

[0074] The pharmaceutical composition of the present invention may contain the active ingredient alone, or may additionally contain one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0075] The carrier, excipient or diluent that may be included in the pharmaceutical composition of the present invention may be, but is not limited to, lactose, dextrose, sucrose, dextrin, maltodextrin, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

[0076]

[0077] The present invention provides a health functional food for preventing or improving atherosclerosis, comprising a beta-agonist or a food-based acceptable salt thereof.

[0078] In the present invention, the type of beta-agonist or a food-based acceptable salt thereof included in the health functional food for preventing or improving atherosclerosis may be the same as the effective ingredient of the pharmaceutical composition for preventing or treating atherosclerosis described above.

[0079] The health functional food of the present invention refers to a food manufactured and / or processed in various forms to provide useful functionality to the human body.

[0080] The health functional food of the present invention can be included in various foods or medicines known in the art.

[0081] There are no specific limitations on the types of foods that can contain the health functional food of the present invention. For example, the health functional food of the present invention can be contained in meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

[0082] The health functional food of the present invention includes all forms such as functional food, nutritional supplement, health food, and food additives, and these types of food can be manufactured in various forms according to conventional methods known in the art. For example, the health food can be manufactured in the form of a liquid drink for consumption, or can be ingested by being granulated, encapsulated, spherical tableted (pills, etc.), or powdered, and can also be manufactured in the form of a powder, capsule, soft capsule, tablet, gum, or adhesive-type liquid composition for consumption. In addition, functional foods include beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meat and their processed foods (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, taffy, dairy products (e.g., butter, cheese, etc.), edible vegetable oils, margarine, vegetable proteins, retort foods, frozen foods, seasonings, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.).

[0083] The health functional food of the present invention may further include ingredients commonly added during food manufacturing, as long as it does not deviate from the ultimate purpose of the present invention, and may further include, for example, proteins, carbohydrates, fats, other nutrients, seasonings, and flavorings.

[0084] The health functional food of the present invention may additionally contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

[0085] The health functional food of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients may be used independently or in combination.

[0086] Ultimately, the present invention provides a pharmaceutical and / or food use of a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof for the treatment, prevention, improvement, and / or inhibition of atherosclerosis. Treatment encompasses the improvement, alleviation, etc. of symptoms associated with atherosclerosis, and prevention encompasses the inhibition of the progression of disease from a pre-disease stage.

[0087] Accordingly, the present invention also provides a method for treating, preventing, improving or inhibiting atherosclerosis, comprising a step of administering a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof to an animal (an animal including a human or an animal other than a human).

[0088] The above animal may be a mammal.

[0089] The above animal may be an animal requiring administration of a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof.

[0090] The above animal may be an animal that has developed or is likely to develop atherosclerosis.

[0091] Additionally, the beta-adrenergic agonist or a pharmaceutically acceptable salt thereof administered above may be an effective amount of a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof.

[0092] The above beta-adrenergic agonist or a pharmaceutically acceptable salt thereof can be appropriately administered to a subject according to a conventional method or administration route used in the art according to the purpose or need, and oral administration is included.

[0093]

[0094] In addition, the present invention provides a use for manufacturing a preparation of a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof for treating or preventing atherosclerosis.

[0095] Unless otherwise stated, the matters mentioned in the compositions, foods, methods, and uses of the present invention apply equally to each other unless they are contradictory.

[0096] The dosage of the beta-agonist, or a pharmaceutically acceptable salt thereof, included in the composition of the present invention or used in the purpose or method is 1 μg / kg / day to 300 μg / kg / day when administered orally, and preferably 1 μg / kg / day to 100 μg / kg / day.

[0097] In addition, an appropriate dosage and frequency of administration can be selected according to a method known in the art, and the amount and frequency of administration of the beta-adrenergic agonist of the present invention or a pharmaceutically acceptable salt thereof actually administered can be appropriately determined by various factors such as the type of symptom to be treated, administration route, sex, health condition, diet, age and weight of the subject, and severity of the disease.

[0098]

[0099] Hereinafter, the present invention will be described in detail with examples to specifically illustrate the invention. However, the following examples are provided merely to facilitate a better understanding of the present invention and are not intended to limit the scope of the present invention.

[0100]

[0101] Example

[0102] [Example 1] Cell preparation

[0103] J774A.1 and RAW264.7 cell lines were cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin-glutamine (PSQ), and LR73 CHO cell line was cultured in alpha-MEM containing 10% FBS and 1% PSQ. Murine bone marrow derived macrophages (BMDMs) were collected from the femoral bone marrow of mice and cultured in RPMI 1640 containing 20% ​​L929 conditioned medium, 10% FBS, and 1% PSQ. THP-1 cell lines were cultured in RPMI 1640 medium containing 10% FBS and 1% PSQ, and differentiated with 100 nM PMA (phorbol-12-myristate-13-acetate) for 48 h before use.

[0104] Mice used in this experiment were purchased from Taconic Biosciences (Germantown, NY, USA) through Damul Science. Mice aged 8–10 weeks were used to obtain BMDM, and 4–6 weeks were used to obtain apoptotic thymocytes. All experiments using mice were approved by the Animal Care and Ethics Committee of the Laboratory Animal Resource Center (LARC) at GIST.

[0105]

[0106] [Example 2] Efferocytosis Assay

[0107] The apoptotic cell removal assay was performed by treating pHrodo (P36600)-stained apoptotic thymocytes with cultured plates containing phagocytes such as J774A.1, RAW264.7, THP-1, or LR-73 for 30 minutes to 2 hours and analyzing them using flow cytometry. Phagocytes were cultured in 24-well culture plates, and one day later, they were treated with ractopamine hydrochloride at 250 nM for 24 hours. Afterwards, pHrodo-stained apoptotic thymocytes were treated at a ratio of 1:20 for 30 minutes to 2 hours. Then, they were washed with pre-chilled PBS, removed from the plates, and measured using a flow cytometer (BD FACS canto II) and analyzed using the equipment's software, FLOWJO (Flowjo LLC).

[0108]

[0109] [Example 3] Apoptotic Cell Binding Assay

[0110] J774A.1 cells were cultured on 18 mm coverglass plates in 12-well plates and treated with 250 nM ractopamine hydrochloride for 24 h. Cy3-labeled beads in cold medium were then added and incubated at 4°C for 30 min. The cells were then washed with PBS, fixed with 4% paraformaldehyde for 15 min, and imaged using an Axio Imager D2 (Zeiss, Jena, Germany). The number of beads bound to phagocytes was then counted.

[0111]

[0112] [Example 4] Time-Lapse Confocal Microscopy

[0113] J774A.1 cells were cultured in 35 mm confocal dishes and treated with 250 nM ractopamine hydrochloride for 24 h. Cells were then stained with Celltraker (CellTracker™ Green CMFDA Dye, C7025, Thermo Fisher) for 30 min or with Lysotracker (Lysotracker Green DND-26, L7526, Invitrogen) for 1 h, followed by treatment with beads stained with Cy3 or Cy5 + pHrodo. Images were then immediately observed using a confocal microscope (FV3000, Olympus) and captured at 10-s intervals for 30 min to 1 h. For internalization studies, the time from target binding to phagocytes until complete incorporation was measured.

[0114]

[0115] [Example 5] Degradation assay

[0116] J774A.1 cells were cultured in 24-well plates and treated with 250 nM ractopamine hydrochloride for 24 h. Afterwards, the cells were treated with Cy3 beads for 15 min, washed with cold PBS, treated with culture medium, and incubated for 30 and 60 min. The cells were then detached and measured using a flow cytometer, and analyzed using the equipment's software, FLOWJO.

[0117]

[0118] [Example 6] Atherosclerotic lesion analysis

[0119] 8-week-old Ldlr for atherosclerosis research - / - Mice were fed a Western diet for 14 weeks. To determine the atherosclerotic-attenuating effect of ractopamine hydrochloride, mice were treated with ractopamine hydrochloride in their drinking water at concentrations of 0.4, 2, and 10 μg / kg for 2–14 weeks while on the Western diet. The control group received vehicle in their drinking water. The drinking water of both the control and experimental groups was replaced once a week. After euthanizing the mice, the heart and artery were harvested and perfused with PBS via the left ventricle. The organs were embedded in frozen sectioning compound (3801480, Leica) and frozen to prepare 7-μm-thick frozen sections. Each section was stained with hematoxylin & eosin (H&E) to count the necrotic core and with Oil red O to measure the plaque lesion area. Tissues stained with each method were photographed using a slide scanner and analyzed.

[0120]

[0121] [Example 7] Statistical Analysis

[0122] Data are presented as the mean ± standard error of the mean (SEM). All experiments were performed independently at least three times, and statistically significant differences were analyzed using an unpaired student's two-tailed t test, one-way ANOVA, or two-way ANOVA using GraghPad Prism 7 software. Significance was considered when the p value was less than 0.05.

[0123]

[0124] Experimental examples 1 to 7

[0125] Experimental examples 1 to 7 were performed using the materials and methods according to the above-described examples.

[0126]

[0127] [Experimental Example 1] Cytotoxicity Test

[0128] The cytotoxicity of ractopamine hydrochloride was confirmed by measuring cell metabolic activity using the MTT assay.

[0129] DMSO or various concentrations of ractopamine hydrochloride were added to J774A.1 macrophages (6x10 3 After incubation for 24 hours with 96 well cells, cell viability was measured using the MTT assay. As shown in Fig. 1, it was confirmed that ractopamine hydrochloride did not affect cell viability.

[0130]

[0131] [Experimental Example 2] Experiment to find the optimal concentration of ractopamine hydrochloride to promote the removal of apoptotic cells.

[0132] Various concentrations of ractopamine hydrochloride were administered to J774A.1 macrophages (4x10 4After incubation with 24 wells (cells / 24 well) for 24 hours, the solution was incubated with pHrodo-stained apoptotic cells for 15 minutes. Then, the level of apoptotic cell removal was measured using flow cytometry. As a result, as shown in Fig. 2, ractopamine hydrochloride 250 nM showed the highest apoptotic cell removal ability. In the following examples, ractopamine hydrochloride 250 nM was used.

[0133]

[0134] [Experimental Example 3] Experiment to evaluate the efficacy of ractopamine hydrochloride in promoting the removal of apoptotic cells.

[0135] Phosphatidylserine (PS) is a representative ligand exposed on the surface of apoptotic cells and recognized by macrophages. PS beads with PS attached to their surface can act as targets for macrophages instead of apoptotic cells.

[0136] The effect of ractopamine hydrochloride on phagocytosis was assessed using PS beads. Macrophages were treated with DMSO or ractopamine hydrochloride for 24 h, then incubated with Cy3-labeled PS beads for 30 min, and phagocytosis was assessed by flow cytometry.

[0137] As a result, as shown in Fig. 3, it was confirmed that ractopamine hydrochloride promotes the removal of apoptotic cells.

[0138]

[0139] [Experimental Example 4] Evaluation of the efficacy of ractopamine hydrochloride in promoting the removal of apoptotic cells in vivo.

[0140] Mice were injected intraperitoneally with ractopamine hydrochloride and incubated for 24 hours. Then, pHrodo-stained apoptotic cells were injected intraperitoneally. After 15 minutes, peritoneal cells were collected and the ability of peritoneal macrophages to remove apoptotic cells was measured using a flow cytometer.

[0141] As a result, as shown in Fig. 4, it was confirmed that ractopamine hydrochloride enhances the apoptotic cell removal ability of macrophages in vivo.

[0142]

[0143] [Experimental Example 5] Experiment to confirm the mechanism of promoting apoptotic cell removal by ractopamine hydrochloride.

[0144] 5-1. Evaluation of the Effect of Ractopamine Hydrochloride on Apoptotic Cell Degradation

[0145] To elucidate the mechanism by which ractopamine hydrochloride promotes the removal of apoptotic cells, we confirmed whether ractopamine hydrochloride increases the internalization of apoptotic cells.

[0146] After treating macrophages with ractopamine hydrochloride for 24 hours, the macrophages were incubated with Cy3-labeled PS beads for 15 minutes, and the Cy3 fluorescence intensity was measured after 30 and 60 minutes using a flow cytometer to evaluate the degree of decrease in Cy3 fluorescence intensity over time.

[0147] As shown in Figure 5, ractopamine hydrochloride exhibited a lower Cy3 mean fluorescent intensity (MFI) compared to the control (DMSO). This confirmed that ractopamine hydrochloride promoted the removal of apoptotic cells by enhancing the degradative capacity of macrophages.

[0148]

[0149] 5-2. Confirmation of increased phagosome-lysosome fusion by ractopamine hydrochloride

[0150] To elucidate the mechanism by which ractopamine hydrochloride increases the macrophage degradation capacity, phagosome-lysosome fusion was measured.

[0151] After treating macrophages with ractopamine hydrochloride for 24 h, phagosome-lysosome fusion was observed and quantified by time-lapse microscopy after incubation of Cy5-labeled PS beads and Lysotracker-stained macrophages.

[0152] As shown in Fig. 6, it was confirmed that fusion signals (lysotracker-pHrodo colocalization signals) increased in ractopamine hydrochloride-treated macrophages.

[0153]

[0154] 5-3. Confirmation of increased lysosome activity due to ractopamine hydrochloride

[0155] Macrophages were treated with ractopamine hydrochloride for 24 hours, stained with Lysotracker, and the Lysotracker intensity was measured and quantified using a confocal microscope.

[0156] As shown in Fig. 7, high Lysotracker fluorescence intensity was confirmed in macrophages treated with ractopamine hydrochloride.

[0157] We confirmed that the increase in fusion signals (lysotracker-pHrodo colocalization signals) in ractopamine hydrochloride-treated macrophages was due to an increase in lysosomes.

[0158]

[0159] [Experimental Example 6] Experiment to confirm the efficacy of ractopamine hydrochloride in an animal model of atherosclerosis.

[0160] To evaluate the effect of ractopamine hydrochloride on atherosclerosis, an atherosclerosis animal model was established using Ldlr knockout mice.

[0161] As shown in Fig. 8, 2-month-old Ldlr KO mice fed a Western-style diet were administered ractopamine hydrochloride (0.4, 2, and 10 μg / kg / day) in drinking water once a week for 13.5 weeks. The arteries were then prepared, and plaque size, necrotic core size, and lipid deposition were determined using H&E and oil red O staining.

[0162] As shown in Figure 9, it was confirmed that the size of plaques was reduced in the ractopamine hydrochloride treatment group compared to the control group.

[0163] Additionally, as shown in Figure 10, it was confirmed that the size of the necrotic core (sky blue dotted line) was reduced in the ractopamine hydrochloride treatment group compared to the control group.

[0164]

[0165] [Experimental Example 7] Experiment to confirm the efficacy of beta-agonists in promoting the removal of apoptotic cells.

[0166] To determine whether beta-agonists other than ractopamine have the effect of promoting apoptotic cell removal, apoptotic cell removal assay was performed using various beta-agonists.

[0167]

[0168] 7-1. Confirmation of the efficacy of salmeterol, salmeterol xinafoate, indacaterol, indacaterol maleate, cloprenaline hydrochloride, and procaterol hydrochloride in promoting the removal of apoptotic cells.

[0169] 100 nM salmeterol, salmeterol xinafoate, indacaterol, indacaterol maleate, cloprenaline hydrochloride and procaterol hydrochloride were administered to J774A.1 macrophages (2x10 4 After incubation with 100 cells / well for 48 hours, the cells were incubated with pHrodo-stained apoptotic cells for 40 minutes. Then, the level of apoptotic cell removal was measured using a flow cytometer.

[0170] As a result, as shown in Fig. 11, it was confirmed that salmeterol, salmeterol xinafoate, indacaterol, indacaterol maleate, cloprenaline hydrochloride, and procaterol hydrochloride promoted the removal of apoptotic cells.

[0171]

[0172] 7-2. Confirmation of the efficacy of fenoterol in promoting the removal of apoptotic cells.

[0173] Fenoterol at a concentration of 100 nM was administered to J774A.1 macrophages (2x10 4After incubation with 100 cells / well for 48 hours, the cells were incubated with pHrodo-stained apoptotic cells for 30 minutes. Then, the level of apoptotic cell removal was measured using a flow cytometer.

[0174] As a result, as shown in Fig. 12, it was confirmed that fenoterol promotes the removal of apoptotic cells.

[0175] This suggests that beta-agonists have preventive or therapeutic effects on atherosclerosis.

Claims

1. A pharmaceutical composition for preventing or treating atherosclerosis, comprising a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof.

2. In claim 1, the beta-agonist is any one selected from the group consisting of the following chemical formulas 1 to 6, a pharmaceutical composition for preventing or treating atherosclerosis: [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] 3. A pharmaceutical composition for preventing or treating atherosclerosis, wherein the prevention or treatment of atherosclerosis according to claim 1 is achieved by promoting efferocytosis.

4. A pharmaceutical composition for preventing or treating atherosclerosis according to claim 1, wherein the prevention or treatment of atherosclerosis is achieved by inhibiting plaque formation.

5. A pharmaceutical composition for preventing or treating atherosclerosis, further comprising a pharmaceutically acceptable carrier, excipient or diluent according to claim 1.

6. A health functional food for preventing or improving atherosclerosis, containing a beta-agonist or a food-based salt thereof.

7. In claim 6, the beta-agonist is any one selected from the group consisting of the following chemical formulas 1 to 6, a health functional food: [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] 8. A health functional food according to claim 6, wherein the prevention or improvement of atherosclerosis is achieved by promoting the removal of apoptotic cells.

9. A health functional food according to claim 6, wherein the prevention or improvement of atherosclerosis is achieved by suppressing plaque formation.

10. A method for preventing, treating, improving or suppressing atherosclerosis by administering a beta-adrenergic agonist or a pharmaceutically acceptable salt thereof to an animal other than a human.

Images