KR20190126214A - Composition for preventing and treating cardiovascular disease comprising lipid extract of Mytilus galloprovincialis - Google Patents

Abstract

The present invention relates to a composition for preventing or treating cardiovascular diseases including Mediterranean mussel lipid extract as an active ingredient and, more specifically, to a pharmaceutical composition for inhibiting vascular inflammation including Mediterranean mussel lipid extract and a sterol compound isolated therefrom. In the composition for preventing or treating cardiovascular diseases, the Mediterranean mussel lipid extract and the active ingredient can be used as a material of medicine and functional food capable of preventing, improving and treating cardiovascular diseases by having stability without causing toxicity to cells as desmosterol, suppressing the expression of cell adhesion molecules, and being excellent in inhibiting vascular inflammation known as the cause of cardiovascular disease.

Description

Composition for preventing and treating cardiovascular disease comprising lipid extract of Mytilus galloprovincialis}

The present invention relates to a composition for the prevention or treatment of cardiovascular diseases comprising a Mediterranean mussel lipid extract as an active ingredient, and more particularly to a pharmaceutical composition for inhibiting vascular inflammation comprising a Mediterranean mussel lipid extract and a sterol compound isolated therefrom. About

The present invention is a cardiovascular disease comprising mediterranean mussel lipid extract and the active ingredient is a Mediterranean mussel lipid extract, characterized in that it has a significant inhibitory effect on the inflammatory response generated in vascular endothelial cells of desmosterol (desmosterol) It relates to a composition for the prophylaxis or treatment of.

Metabolic syndromes such as hypertension, hyperlipidemia, and diabetes caused by overnutrition, imbalance, lack of exercise, and stress are major risk factors for atherosclerosis, and mortality due to cardiovascular disease caused by an increase in these diseases. Increasingly, social problems are emerging (Lee et al., Int. J. Mol. Sci 13, p5628-5644, 2012). In particular, atherosclerosis of representative vascular disease is a major cause of myocardial infarction and stroke, the leading cause of death worldwide (Funatsu et al., Jpn. J. Ophthalmol. 45, p577-584, 2001).

Atherosclerosis is an inflammatory disease of vascular endothelial cells and is the most common pathological condition of cardiovascular disease. Adhesion of vascular endothelial cells by stimulation of proinflammatory cytokines with monocytes or macrophages circulating in the bloodstream indicates the onset of atherosclerosis (Packard and Libby, Clin Chem 54, 24-38. 2008 ). The onset and progression of atherosclerosis has recently been reported to be caused by chronic inflammation (Packard and Libby, Clin Chem 54, 24-38. 2008; Madge et al., Exp. Mol. Pathol. 70, 317-325, 2001).

Attachment to vascular endothelial cells of monocytes is primarily regulated by intracellular signaling promoted by chemokines such as interleukin (IL) 8 and monocyte chemoattractant protein (MCP) 1. In vascular endothelial cells of monocytes, adhesion is also regulated by endothelial cell adhesion factors such as intercellular adhesion molecule (ICAM) 1 and vascular adhesion molecule (VCAM) 1 (Betera et al., Rev. Neuropsiquiatr. 37, 177- 195). MCP-1 acts as a chemoattractant for macrophages and monocytes, and IL-8 acts as a chemoattractant for neutrophils and plays an important role in the initiation and progression of atherosclerosis (Sprague and Khalil, Biochem. Pharmacol., 78, 539). -552, 2009; Yang et al., Ann. Rheum. Dis., 63, 1511-1513, 2004). Cholesterol deposits in the lining of blood vessels, and when endothelial cells proliferate, angiomas are formed, which narrow the vessel's lumen and form blood clots, and other heart diseases such as angina, myocardial infarction, endocarditis, and arrhythmia, and cerebral vessels such as cerebral infarction and cerebral hemorrhage. Diseases and other complications such as kidney failure and ischemic limb disease (Lee et al., Inflammation 35, 584-93. 2012; Cho et al., Korean J. Plant Res 29, 525-531. 2016) .

TNF-α (Tumor Necrosis Factor-α) is an atherosclerotic activity and nuclear factor kappa B (NF-κB) is a key pro-inflammatory cytokine in the inflammatory cascade, which plays an important role in vascular inflammation and induces atherosclerosis. The transcription factor NF-κB up-regulates chemotactic cytokines, including adhesion molecules such as VCAM-1 and ICAM-1 and MCP-1 (Boyle et al., Circulation, 98, 282-288, 1998). Nallasamy et al., J. Nutr. Biochem., 25, 824-833, 2014). Therefore, the regulation of TNF-a-induced inflammation is an important indicator of alleviation of vascular inflammatory diseases such as atherosclerosis.

Therefore, due to the increasing incidence and mortality of cardiovascular disease, a lot of medical attention and research is progressing, the treatment of non-causing side effects with the more fundamental treatment is the urgent development of such therapeutics.

The Mediterranean Sea ( Mytilus galloprovincialis ) is native to Europe, the coast of the Atlantic Ocean, the Mediterranean Sea, Hong Kong, southern Japan, Korea, and all over the world. It is usually attached to the rock or solid material of intertidal zone as a foot bark. It is a two-year-old, and is used for food as a large-scaled species in Korea.

Antibiotic, antifungal, antimicrobial, antiprotozoal and antiviral effects of mediterranean mussels using proteolytic enzymes and peptide components isolated therefrom are known (Hubert et al., Euro. J. Biochem., 240, 302- 306, 1996; Mitta et al., J. Cell Sci., 112, 4233-4242, 1999; Roch et al., Evidenced-based Complementary and Alternative Medicine, 1, 167-174, 2004; Romestand et al., Euro J. Biochem., 270, 2805-2813, 2004), the association of cardiovascular disease prevention due to vascular inflammation is unknown.

Therefore, in the present invention, attention was focused on mediterranean mussel lipid extract as a material for new cardiovascular treatment, Mediterranean mussel is indigenous to Europe, the coast of the Atlantic, Mediterranean, Hong Kong, southern Japan, Korea, etc. A two-year-old who lives in the form of a horticultural attachment to an indigo plant and is used for food as a large-sized farming species in Korea.

Antibiotic, antifungal, antibacterial, antiprotozoal, and antiviral effects of hydrolyzate using protease and peptide components isolated from mediterranean mussels are known, but there is no known association with cardiovascular disease prevention due to vascular inflammation. .

It is therefore an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of cardiovascular diseases comprising a lipid extract of mediterranean mussels as an active ingredient.

Another object of the present invention to provide a functional health food for the prevention or improvement of cardiovascular disease comprising mediterranean mussel lipid extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cardiovascular diseases comprising desosterol as an active ingredient.

In addition, another object of the present invention is to provide a functional health food for the prevention or improvement of cardiovascular disease comprising des-sterol as an active ingredient.

In order to achieve the object of the present invention as described above, the present invention is a pharmaceutical composition for the prevention or treatment of cardiovascular diseases comprising mediterranean mussel lipid extract as an active ingredient is a blood vessel containing a mediterranean mussel lipid extract and a sterol compound separated therefrom It relates to a pharmaceutical composition for inhibiting inflammation,

The mediterranean mussel lipid extract of the present invention is a lipid extract obtained by the supercritical extraction method and the active ingredient contains a desmosterol compound, does not cause toxicity to the cell and is stable, expression of cell adhesion molecules Mediterranean mussel lipid extract, characterized in that it can inhibit the vascular inflammation known to be the cause of cardiovascular disease, and can be used as a material of medicines and functional foods that can prevent, improve and treat cardiovascular diseases. To provide a composition for the prevention or treatment of cardiovascular diseases comprising as an active ingredient.

The present invention also provides a functional health food for the prevention or improvement of cardiovascular disease comprising mediterranean mussel lipid extract as an active ingredient, and the desostrol compound is isolated and purified from mediterranean mussel.

The present invention relates to a pharmaceutical composition for preventing or treating cardiovascular diseases comprising mediterranean mussel lipid extract as an active ingredient, wherein the composition is toxic to cells as a composition comprising meditated mussel lipid extract and a compound isolated from the extract as an active ingredient. It is stable and does not induce the expression of cell adhesion molecules, and it has an excellent effect of inhibiting vascular inflammation known as the cause of cardiovascular disease, and it is a medicine and functional food that can prevent, improve and treat cardiovascular disease. It can be used as a material.

1 is a measure of mediterranean mussel lipid extract cytotoxicity of the present invention
Figure 2-4 is a measure of the inhibitory effect of cell adhesion factor expression of mediterranean mussel lipid extract of the present invention
5 is a measure of the inhibitory effect of nitric oxide (NO) production of mediterranean mussel lipid extract of the present invention
6-9 is a measure of the inhibitory effect of the inflammatory cytokines of the Mediterranean mussel lipid extract of the present invention
10 is a measure of the inhibitory effect of expression of MMP-9 by mediterranean mussel lipid extract of the present invention
12 is a cytotoxic effect analysis measurement of the des Mosterol of the present invention
Figure 13-15 is a measurement of the cell adhesion factor expression inhibitory effect of the des Mosterol of the present invention
Figure 16 is a measurement of the effect of inhibiting the production of Nitric oxide (NO) of the desosterol of the present invention
17-20 is a measure of the inhibitory effect of the inflammatory cytokines of the destestolol of the present invention.

The present invention is characterized in that mediterranean mussel lipid extract can prevent or treat cardiovascular diseases.

The present invention relates to a composition for the prevention or treatment of cardiovascular diseases comprising a Mediterranean mussel lipid extract as an active ingredient, and more particularly to a pharmaceutical composition for inhibiting vascular inflammation comprising a Mediterranean mussel lipid extract and a sterol compound isolated therefrom. The mediterranean mussel lipid extract and the active ingredient is desostosterol (desmosterol) does not cause toxicity to the cell is stable, can inhibit the expression of cell adhesion molecules, and known vascular inflammation as a cause of cardiovascular disease In the composition for the prevention or treatment of cardiovascular diseases, including Mediterranean mussel lipid extract as an active ingredient, which is excellent in the suppressed effect, which can be used as a material of medicines and functional foods that can prevent, improve and treat cardiovascular diseases. It is about.

Therefore, below, preferred embodiment of this invention is described in detail with reference to an Example. However, the scope of the present invention is not limited to these examples.

Example 1 Preparation of Mediterranean Sea Mulberry Lipid Extract

Mediterranean mussels used in the present invention were purchased from Masanhappo-gu, Changwon-si, and Mediterranean mussel lipid extracts were obtained by the following method. First, mediterranean mussels were prepared by lyophilization of mediterranean mussels for extraction, and then 2.0 kg of the powder obtained by grinding was extracted with a carbon dioxide supercritical extraction method at a temperature of 50 ° C., a pressure of 400 bar and a time of 120 minutes to obtain a lipid extract of about 70 g. Taking a part, the following experiment was performed.

Experimental Example 1 Analysis of Cytotoxic Effects of Mediterranean Sea-Solid Lipid Extracts

Mediterranean mussel lipid extracts obtained in Example 1 were measured.

In order to confirm the cytotoxicity of the mediterranean mussel lipid extract, vascular endothelial cells were cultured at 37 ° C. and 5% CO ₂ using EGM ^TM -2 Medium (Endothelial Growth Medium-2) containing 2% FBS and VEGF. It was. Cell survival rate was measured using WST-1 in vascular endothelial cells. After dispensing HUVEC cells into 5 × 10³cells / well in a 96 well plate, treat the samples by concentration and incubate for 40 hours. The absorbance was measured at 450 nm using an ELISA reader (US / MQX 200; Bio-Tek Instruments Inc., Winooski, VT, USA). The measured value was analyzed as the average value of three replicate experiments.

As a result, as shown in FIG.

Cytotoxicity of methanol lipid extract against HUVEC cells did not show up to 200 μg / mL concentration.

Therefore, the lipid extract used in the present invention was found to be stable without causing any particular toxicity to the cells.

Experimental Example 2 Inhibitory Effect of Mediterranean Sea Soil Lipid Extracts on Cell Attachment Factor

High concentrations of cholesterol in the blood are deposited by lipoproteins, extracellular matrix, calcium and immune cells by vascular endothelial cells through the mediator of cell adhesion factors. In order to measure the inhibitory effects of VCAM-1, ICAM-1, and E-selectin on the adhesion of mediterranean-seasoned lipid extracts involved in adhesion of vascular endothelial cells of monocytes, vascular endothelial cells were placed on 6 well plates for 24 hours. After incubation, mediterranean mussel lipid extracts were treated by concentration, and after 2 hours, TNF-α (20 ng / mL) was treated and cultured for 40 hours. The supernatants were collected and ELISA kit (R & D systems, Inc., Minneapolis, MN, USA) was used to express the expression levels of VCAM-1, ICAM-1, and E-selectin in ELISA reader (US / MQX 200; Bio-Tek Instruments Inc.). , Winooski, VT, USA) at 450 nm.

As a result, as shown in Figs. 2-4, mediterranean mussel lipid extracts were cell adhesion factors VCAM-1 (see Fig. 2), ICAM-1 (see Fig. 3), and E-selectin (Fig. 3) at a concentration of 100 μg / mL. 4) showed a concentration-dependent significant inhibition.

Therefore, mediterranean mussel lipid extract inhibited the adhesion of monocytes to endothelial cells by inhibiting adhesion factors such as ICAM-1, VCAM-1, and E-selectin, and inhibited the migration of monocytes to vascular endothelium. It was found that it can be suppressed.

Experimental Example 3 Inhibition Effect of Nitric Oxide (NO) Formation of Mediterranean Sea-Soaked Lipid Extracts

In order to measure the inhibitory effect of nitric oxide (NO) production, which is an inflammatory mediator of mediterranean mussel lipid extracts, vascular endothelial cells were cultured in 6 well plates for 24 hours, and methane lipid extracts were treated in different concentrations. After treatment with α (20 ng / mL) it was incubated for 40 hours. After removing the medium, 120 μL of lysis buffer and scraping the cells, centrifuged for 10 minutes at 14,000 rpm. After 100 μL of the supernatant and 100 μL of Griess reagent were reacted in the dark for 20 minutes, the absorbance was measured at 540 nm. NO concentration was determined using the concentration-specific standard curve of the nitrite standard.

As a result, as shown in Figure 5, mediterranean mussel lipid extract showed a concentration-dependent significant inhibition of the production of inflammatory reaction mediator Nitric oxide (NO) from the concentration of 100 μg / mL. Therefore, it can be seen that mediterranean mussel lipid extracts can suppress vascular inflammation by inhibiting inflammatory mediators.

Experimental Example 4 Inhibitory Effect of Mediterranean Sea-Solid Lipid Extracts on Pro-inflammatory Cytokines

Cytokines, substances that induce inflammatory cells to move into blood vessels and induce the action of inflammatory cells, are interleukin-6 (IL-6), interleukin-1β (IL-1β), interleukin-8 (IL-8), and MCP-1. Etc. IL-6 and IL-1β are secreted in the early stages of inflammation and cause various diseases including inflammatory diseases during overproduction. IL-8 is a neutrophil chemoattractant, and MCP-1 (monocyte chemoattractant protein-1) is a macrophage and mononuclear. To measure the inhibitory effect of mediterranean-branched lipid extract on the expression of cytokine involved in vascular inflammation, which acts as a chemoattractant for cells and plays an important role in the initiation and progression of atherosclerosis,

After culturing vascular endothelial cells in 6 well plates for 24 hours, mediterranean mussel lipid extracts were treated by concentration, and 2 hours later, TNF-α (20 ng / mL) was treated. After incubation for 40 hours, the supernatants were collected and ELISA kit (R & D systems, Inc., Minneapolis, MN, USA) was used to measure the expression levels of IL-6, IL-8, and MCP-1 using ELISA reader (US / MQX 200 (Bio-Tek Instruments Inc., Winooski, VT, USA) at 450 nm.

As a result, as shown in Figs. 6-9, mediterranean mussel lipid extracts express the expression of inflammatory cytokines IL-6 (see Fig. 6) and IL-1β (see Fig. 7) at a concentration of 100 μg / mL. 8 (see FIG. 8) showed a concentration-dependent significant inhibition from 150 μg / mL concentration, but did not significantly inhibit the expression of MCP-1 (see FIG. 9). Thus, it was found that mediterranean mussel lipid extracts can inhibit vascular inflammation by inhibiting the expression of inflammatory cytokines, which induce inflammatory cells to move into blood vessels and induce the action of inflammatory cells.

Experimental Example 5 Inhibitory Effect of MMP-9 Expression by Mediterranean Sea Soil Lipid Extract

The transcriptional activity of NF-κB is known to regulate not only proinflammatory cell adhesion molecules but also the expression of matrix metalloproteinase (MMP) -9, called gelatinase. MMP-9 plays an important role in the migration, extravasation and infiltration of immune cells, including monocytes. After culturing vascular endothelial cells in 6 well plates for 24 hours, mediterranean mussel lipid extracts were treated by concentration, and 2 hours later, TNF-α (20 ng / mL) was treated. After incubation for 40 hours, the supernatants were collected and ELISA kit (R & D systems, Inc., Minneapolis, MN, USA) was used to measure the expression level of MMP-9 using ELISA reader (US / MQX 200; Bio-Tek Instruments Inc., Winooski, VT, USA) at 450 nm.

As a result, as shown in Figure 10, mediterranean mussel lipid extract showed a concentration-dependent significant inhibition of MMP-9 involved in the migration, extravasation, infiltration of immune cells including monocytes from the concentration of 100 μg / mL . Therefore, it was found that mediterranean mussel lipid extracts can inhibit vascular inflammation by inhibiting immune cell migration, extravasation and invasion.

Example 2 Identification of Compound

Mediterranean mussel lipid extracts were qualitatively analyzed using a gas chromatography mass spectrometer (GC-MS, Shimadzu QP-2010 Ultra), and the compounds of desostrol were identified.

Experimental Example 6

Desmosterol and TNF-α were purchased from Sigma Chemical Company (St. Louis, MO, USA). Endothelial cells (HUVECs) were purchased from Lonza (Walkersville, MD, USA), Cell Proliferation Reagent WST-1 for Roche (Mannheim, Germany) and ELISA kit for R & D systems, Inc. Purchased from Minneapolis, MN, USA.

<Formula 1>

Desmosterol (5,24-cholestadien-3β-ol)

Experimental Example 7 Analysis of Cytotoxic Effects of Desmosterol

Cytotoxicity was measured by treating the desostosterol obtained in Example 2 to HUVEC, which is a vascular endothelial cell.

In order to confirm the cytotoxicity of the sample, cell viability was measured using WST-1 in cultured vascular endothelial cells. Dispense the HUVEC cells into a 96 well plate at 5 × 10³ cells / well, incubate for 40 hours with desmosterol concentration, and treat 10 μL of WST-1 solution for each well. After the reaction, the absorbance at 450 nm was measured using an ELISA reader (US / MQX 200; Bio-Tek Instruments Inc., Winooski, VT, USA). The measured value was analyzed as the average value of three replicate experiments.

As a result, as shown in Figure 12, the cytotoxicity of Desmosterol against HUVEC cells did not appear up to 250 μM concentration. Therefore, it can be seen that the desmosterol used in the present invention is stable without causing specific toxicity to cells.

Experimental Example 8 Inhibitory Effect of Desmosterol on Cell Attachment Factor Expression

The high concentration of cholesterol in the blood is mediated by cell adhesion factors, and lipoproteins, extracellular matrix, calcium and immune cells are attached to vascular endothelial cells. In order to measure the inhibitory effect of the expression of VCAM-1, ICAM-1, and E-selectin, the cell adhesion factors of Desmosterol involved in the adhesion of vascular endothelial cells of mononuclear cells, 24 vascular endothelial cells were placed on 6 well plates. After incubation for a time, desosterol was treated by concentration, and after 2 hours, TNF-α (20 ng / mL) was treated and then incubated for 40 hours. The supernatants were collected and ELISA kit (R & D systems, Inc., Minneapolis, MN, USA) was used to express the expression levels of VCAM-1, ICAM-1, and E-selectin in ELISA reader (US / MQX 200; Bio-Tek Instruments Inc.). , Winooski, VT, USA) at 450 nm.

As a result, as shown in Figs. 13-15,

Desmosterol showed a concentration-dependent significant inhibition of the expression of cell adhesion factors VCAM-1 (see FIG. 13), E-selectin (see FIG. 15) from 31.3 μM concentration, and ICAM-1 (FIG. Expression showed significant inhibitory activity at concentrations above 125 μM.

Therefore, desmosterol inhibited the adhesion of monocytes to endothelial cells by inhibiting adhesion factors such as ICAM-1, VCAM-1, and E-selectin, and inhibited the migration of monocytes to vascular endothelial blood vessels. It was found that inflammation can be suppressed.

Experimental Example 9 Inhibitory Effect of Desmosterol on Nitric Oxide (NO) Formation

In order to measure the inhibitory effect of desmosterol on nitric oxide (NO) production, an inflammatory mediator, vascular endothelial cells were cultured in 6 well plates for 24 hours. 40 hours after treatment with -α (20 ng / mL). After removing the medium, 120 μL of lysis buffer and scraping the cells, centrifuged for 10 minutes at 14,000 rpm. After 100 μL of the supernatant and 100 μL of Griess reagent were reacted in the dark for 20 minutes, the absorbance was measured at 540 nm. NO concentration was determined using the concentration-specific standard curve of the nitrite standard.

As a result, as shown in Figure 16, Desmosterol showed a concentration-dependent significant inhibition of the production of inflammatory mediator NO from 31.3 μM concentration. Therefore, it was found that desosterone can suppress vascular inflammation by inhibiting inflammatory mediators.

Experimental Example 10 Inhibitory Effect of Desosterol on Pro-inflammatory Cytokines

Cytokines, substances that induce inflammatory cells to move into blood vessels and induce the action of inflammatory cells, are interleukin-6 (IL-6), interleukin-1β (IL-1β), interleukin-8 (IL-8), and MCP-1. Etc. IL-6 and IL-1β are secreted in the early stages of inflammation and cause various diseases including inflammatory diseases during overproduction. IL-8 is a neutrophil chemoattractant, and MCP-1 (monocyte chemoattractant protein-1) is a macrophage and mononuclear. Vascular endothelial cells were cultured in 6 well plates for 24 hours to measure the inhibitory effect of destestolol on the expression of cytokine involved in vascular inflammation, which plays an important role in the initiation and progression of arteriosclerosis by acting as a chemoattractant of cells. After, mediterranean mussel lipid extract was treated by concentration and treated with TNF-α (20 ng / mL) after 2 hours. After incubation for 40 hours, the supernatants were collected and ELISA kit (R & D systems, Inc., Minneapolis, MN, USA) was used to measure the expression levels of IL-6, IL-8, and MCP-1 using ELISA reader (US / MQX 200 (Bio-Tek Instruments Inc., Winooski, VT, USA) at 450 nm.

As a result, as shown in Figs. 17-20,

Desosterol showed inflammatory cytokines IL-6 (see FIG. 17) and MCP-1 (see FIG. 20) at a concentration of 31.3 μM, showing a concentration-dependent significant inhibition, IL-1β (see FIG. 18) and IL -8 (see Figure 19) showed a concentration-dependent significant inhibition from 62.5 μM concentration.

Therefore, it has been found that desosterone can inhibit vascular inflammation by inhibiting the expression of a substance inflammatory cytokine that induces inflammatory cells to move into blood vessels and induces the action of inflammatory cells.

Therefore, according to the embodiment and the experimental results,

In the present invention, it can be confirmed through various experiments that mediterranean mussel lipid extracts can treat cardiovascular diseases, and in particular, blood vessels are mainly composed of endothelial cells and smooth muscle cells, and inflammatory cells such as monocytes circulating in the early stage of inflammation are endothelial. Monocytes are differentiated into macrophages by cells moving into the blood vessel wall. For this purpose, monocytes are attached to the surface of endothelial cells to express cell adhesion molecules such as ICAM-1 or VCAM-1.

In addition, TNF-α (Tumor Necrosis Factor-α) is an inflammatory mediator cytokine known to play an important role in systemic inflammation and immune response, and induces the expression of these adhesion molecules in damage of cardiovascular diseases including atherosclerosis.

In addition, the Mediterranean mussel lipid extract according to the present invention effectively inhibited the inflammatory cytokine production and expression and activity of ICAM-1 or VCAM-1 by TNF-α composition comprising mediterranean mussel lipid extract as an active ingredient is a cardiovascular disease Can be effectively prevented or treated.

In the present invention, "cardiovascular disease" includes hypertension, arrhythmia, heart failure, heart attack, myocardial infarction, arteriosclerosis, angina pectoris, stroke, cerebral hemorrhage, hemangioma, coronary artery disease, aortic disease, cerebrovascular disease, ischemic disease and peripheral vascular disease You can,

In the present invention, mediterranean mussel lipid extract may be obtained by using a supercritical extraction method, 'lipid extract' defined in the present invention includes that extracted from the mediterranean sea mussel using a supercritical extraction method including carbon dioxide supercritical extraction method do.

The composition of the present invention comprising the mediterranean mussel lipid extract as an active ingredient can be used as a pharmaceutical composition or a food composition, the pharmaceutical composition is prepared using a pharmaceutically acceptable and physiologically acceptable adjuvant in addition to the active ingredient. Excipients, disintegrants, sweeteners, binders, coatings, swelling agents, lubricants, lubricants or flavoring agents may be used as the adjuvant.

In addition, the pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the active ingredient described above through administration, wherein the pharmaceutical form is a capsule Preparations, liquids, syrups, suspensions, emulsions, drops, granules, powder tablets, coated tablets or liquids available for injection. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, nontoxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include but are not limited to natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, corn sweeteners, acacia, trackercance or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers in compositions formulated in liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can be formulated according to each disease or component, as appropriate in the art.

In one embodiment of the invention, the composition of the present invention may also be a food composition,

Mediterranean mussel lipid extract of the present invention may be included in the composition at a concentration of 10 to 300μg / mL or more, and also the Mediterranean mussel lipid extract of the present invention may be included in 0.1 to 95% by weight relative to the total weight of the composition. In particular, mediterranean mussel lipid extract according to the present invention was confirmed through one embodiment that it does not induce toxicity to cells.

Therefore, the food composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as well as the conventional Mediterranean food composition, in addition to the mediterranean mussel lipid extract as an active ingredient. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The aforementioned flavoring agents can advantageously be used natural flavoring agents (tautin), stevia extracts (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).

The food composition of the present invention may be formulated in the same manner as the pharmaceutical composition, used as a functional food, or added to various foods. Foods to which the composition of the present invention may be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice creams, alcoholic beverages, vitamin complexes, and health supplements. There is this.

In addition, the food composition is a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, such as Mediterranean soybean lipid extract as an active ingredient, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and Salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain a fruit flesh for producing natural fruit juice and fruit juice beverage and vegetable beverage.

Mediterranean mussel lipid extract, an active ingredient of the present invention, is a natural substance and has almost no side effects such as chemicals, and thus can be used safely even when taken for a long time for the purpose of preventing or treating inflammatory diseases.

The present invention also provides a health functional food for improving cardiovascular disease comprising mediterranean mussel lipid extract as an active ingredient.

The health functional food of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like for the purpose of improving inflammatory diseases.

In the present invention, "health functional food" refers to a food prepared and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, and nutrients for the structure and function of the human body. It is meant to be consumed for the purpose of regulating or obtaining a useful effect for health use such as physiological action.

The health functional food of the present invention may include a conventional food additive, and the suitability as a food additive, unless otherwise specified, in accordance with the General Regulations of the Food Additives and General Test Methods approved by the Food and Drug Administration, etc. Judging by the standards and standards.

Examples of the items listed in the "Food Additive Reduction" include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid and cinnamic acid; Natural additives such as dark blue pigment, licorice extract, crystalline cellulose, high color pigment and guar gum; Mixed preparations, such as a sodium L- glutamate preparation, a noodles addition alkali preparation, a preservative preparation, and a tar pigment preparation, etc. are mentioned.

For example, a dietary supplement in the form of tablets is a mixture of the Mediterranean seaweed lipid extract, an active ingredient of the present invention, with excipients, binders, disintegrants and other additives, and then granulated in a conventional manner, and then a lubricant is added. Compression molding or the mixture can be compression molded directly. In addition, the health functional food in the form of tablets may contain a mating agent or the like as necessary.

Hard capsules of the health functional foods in the form of capsules can be prepared by filling a mixture of a mixture of additives such as mediterranean mussel lipid extract, which is the active ingredient of the present invention, into a conventional hard capsule, and soft capsules are mediterranean mussel lipid extracts. The mixture mixed with additives such as excipients can be prepared by filling in a capsule base such as gelatin. The soft capsule agent may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, as necessary.

The health functional food in the form of a cyclic form may be prepared by molding a mixture of a Mediterranean soybean lipid extract, an active ingredient of the present invention, an excipient, a binder, a disintegrant, and the like by using a conventionally known method. It can be zeroed away or the surface can be coated with materials such as starch, talc.

The health functional food in the form of granules can be prepared by granulating a mixture of the Mediterranean sea urine lipid extract and the excipient, the binder, the disintegrant, etc., which are the active ingredients of the present invention, and, if necessary, flavoring agent, copulation, It may contain an agent and the like.

The health functional food may be beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements.

The present invention also provides a method for preventing or treating cardiovascular disease, comprising administering mediterranean mussel lipid extract to a mammal.

As used herein, the term "mammal" refers to a mammal that is the subject of treatment, observation or experiment, preferably human.

As used herein, the term “therapeutically effective amount” means an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as thought by a researcher, veterinarian, doctor or other clinician, which Amounts that induce alleviation of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dosages and frequency of administrations for the active ingredients of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be readily determined by one skilled in the art and includes the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, general health of the patient. , Sex and diet, time of administration, route of administration and rate of composition, duration of treatment, and drugs used concurrently. In the treatment method of the present invention, in the case of an adult, when mediterranean mussel lipid extract of the present invention is administered once to several times a day, it is preferable to administer at a dose of 0.01 mg / kg to 250 mg / kg.

In the treatment method of the present invention, the composition comprising the mediterranean-sealed lipid extract of the present invention as an active ingredient may be administered by oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. Can be administered in a conventional manner.

In addition, the present invention can provide a pharmaceutical composition for the prevention or treatment of cardiovascular diseases, using the desmosterol compound isolated and purified from mediterranean mussel lipid extract as well as mediterranean mussel lipid extract.

The desmosterol compound according to the present invention may be used in the form of a salt, preferably a pharmaceutically acceptable salt. As the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is preferable, and an organic acid and an inorganic acid may be used as the free acid. The organic acid is not limited thereto, citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, Glutamic acid and aspartic acid. In addition, the inorganic acid includes, but is not limited to, hydrochloric acid, bromic acid, sulfuric acid and phosphoric acid.

Desmosterol compounds according to the present invention may be isolated from nature or prepared by chemical synthesis known in the art, preferably may be isolated and purified from mediterranean mussel lipid extract. Separation and purification of these compounds from the extract is performed alone or in combination with column chromatography and high performance liquid chromatography (HPLC) filled with various synthetic resins such as silica gel or activated alumina. Can be used. However, the extraction and separation and purification method of the active ingredient is not necessarily limited to the above method.

Claims (7)

Mediterranean mussels ( Mytilus galloprovincialis ) A pharmaceutical composition for the prevention or treatment of cardiovascular diseases comprising a lipid extract as an active ingredient. The method of claim 1,
Pharmaceutical composition for the prevention or treatment of cardiovascular diseases comprising desmosterol (Desmosterol) isolated from the Mediterranean extract ( Mytilusgalloprovincialis ) lipid extract.
The method of claim 2,
Desmosterol (Desmosterol) is a pharmaceutical composition for the prevention or treatment of cardiovascular diseases, characterized in that represented by the formula (1).
[Formula 1]

The method of claim 2,
Desmosterol (Desmosterol) is a pharmaceutical composition for the prevention or treatment of cardiovascular diseases, characterized in that separated using a supercritical extraction method.
The method of claim 1,
Cardiovascular disease is selected from the group consisting of hypertension, arrhythmia, heart failure, heart attack, myocardial infarction, arteriosclerosis, angina, stroke, cerebral hemorrhage, hemangioma, coronary artery disease, aortic disease, cerebrovascular disease, ischemic disease and peripheral vascular disease A pharmaceutical composition for the prevention or treatment of cardiovascular diseases.
The method of claim 1,
Desmosterol (Desmosterol) is a pharmaceutical composition for the prevention or treatment of cardiovascular diseases, characterized in that to inhibit vascular inflammation in vascular endothelial cells.
Food composition for the prevention or improvement of cardiovascular diseases comprising desmosterol isolated from the Mediterranean extract ( Mytilusgalloprovincialis ) lipid extract.

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