KR20110063129A - A pharmaceutical composition for preventing or treating hypercholesterolemia or cardiovascular disease and a health functional food for preventing or improving hypercholesterolemia or cardiovascular disease, containing an extract of hericium erinaceum hypha cultivated with artemisia capillaries as an effective ingredient - Google Patents

Abstract

PURPOSE: A composition containing Hericium erinaceum mycelium extract is provided to prevent or treat hyperlipidemia or cardiovascular diseases without side effects. CONSTITUTION: A pharmaceutical composition for preventing or treating hyperlipidemia or cardiovascular diseases contains Hericium erinaceum mycelium cultured in Artermisia capillaris medium and solvent extract of the mixture of the Artermisia capillaris medium as an active ingredient. The Artermisia capillaris medium additionally contains rice bran. The extraction solvent is water, alcohol of 1-5 carbon atoms, hexane, chloroform, methylene chloride, acetonitrile, acetone, ethyl acetate.

Description

A pharmaceutical composition for the prevention or treatment of hyperlipidemia or cardiovascular disease, and the health functional food for the prevention or improvement of hyperlipidemia or cardiovascular disease, containing the extract of the scaber mycelium mycelium mycelium cultured in Injin mugwort medium as an active ingredient hypercholesterolemia or cardiovascular disease and a health functional food for preventing or improving hypercholesterolemia or cardiovascular disease, containing an extract of Hericium erinaceum Hypha cultivated with Artemisia capillaries as an effective ingredient}

The present invention relates to a pharmaceutical composition for the prevention or treatment of hyperlipidemia or cardiovascular disease, and to the health functional food for the prevention or improvement of hyperlipidemia or cardiovascular disease, containing the extract of the Roeden mushroom mycelium cultured in Injin mugwort medium as an active ingredient.

Hyperlipidemia refers to a condition where the amount of fat in the blood is higher than normal. Too much fat in the blood deposits a mass of fat on the walls of blood vessels, and over time the fat mass causes a condition called atherosclerosis, which narrows the inner diameter of the arteries and blocks blood flow. As a result, stenosis occurs in the coronary artery, an artery that supplies blood to the heart muscle, and the heart fails to receive enough oxygen and nutrients, resulting in coronary artery disease such as angina pectoris or myocardial infarction. Stroke is induced, and less blood flow to the lower limbs, necrosis of the limbs is caused by a lack of blood supply. Such hyperlipidemia can be classified into hypercholesterolemia, hypertriglyceridemia, low density lipoprotein cholesterolemia and the like.

The cardiovascular disease caused by the hyperlipidemia is the leading cause of death in Korea, which continues to increase due to lack of exercise, stress, and eating out of modern people. The average blood cholesterol level in Koreans was 60s (139-166mg / dl), 70s (154-189mg / dl), 80s (175mg / dl), 90s (184mg / dl), 2000s (203mg / dl) This is increasing year by year, similar to the increase in the incidence of cardiovascular disease.

Various drugs have been developed for the treatment of hyperlipidemia. Blood cholesterol reduction was 5-10% in pyruvate derivatives, 15-20% in nicotinic acid derivatives and 15-25% in bile acid-binding resins, whereas HMG CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase inhibitors were 25 -45% showed higher activity than other drugs. Statin-based drugs, HMG CoA reductase inhibitors, are widely used as the first drug for people with hypercholesterolemia. Depending on the formulation, they lower blood levels of LDL, the primary target for treating hyperlipidemia, by 18-55%. Known. Statins are structural analogs of HMG CoA, an intermediate of cholesterol biosynthesis, which inhibit the activity of HMG CoA reductase, which catalyzes the reduction of HMG CoA to mevalonic acid during the cholesterol biosynthesis process. These drugs include lovastatin, pravastatin, simvastatin, fluvastatin, cerivastatin, atorvastatin, rosuvastatin and pitavastatin (pitavastatin) pitavastatin). According to 4S-study (Scandinavian Simvastatin Survival Study Group, 1994), 4444 patients with a history of angina or myocardial infarction were treated with 20-40 mg of simvastatin for 5.4 years, resulting in total cholesterol, LDL-cholesterol, and HDL- Cholesterol levels varied by -25%, -37%, and 8%, respectively, and deaths from coronary artery disease decreased by 42%. In addition, when comparing atorvastatin and other HMG CoA reductase inhibitors, total cholesterol was lower than atorvastatin compared to pravastatin (-16%), lovastatin (-18%), simvastatin (-21%), and placebo (1%). -29% showed a significant reduction compared to other drugs, LDL-cholesterol compared to lovastatin (-21%), pravastatin (-23%), simvastatin (-26%), placebo (-1%) Statins showed a further decrease of -37%. In a domestic study of HMG CoA reductase inhibitors, pravastatin showed a change of -17 to -23% in total cholesterol and -24 to -34% in LDL, lovastatin -30.9% in total cholesterol, and -34% in LDL. There was a change. Both lovastatin and simvastatin are effective in improving lipid levels in hyperlipidemia. Simvastatin has a similar effect with 50% dose of lovastatin, and atorvastatin significantly reduces serum total cholesterol, LDL and triglycerides in Koreans. There was a clinical report. In addition to statin drugs, ethyl linoleate is currently sold as a treatment for hyperlipidemia. However, in 2001, serious side effects, such as rhabdomyolysis and myopathy, resulted in increased interest in the side effects of these statin preparations after cerivastatin was canceled by the US Food and Drug Administration. Common side effects of statin preparations include gastrointestinal symptoms, myalgias, rare side effects such as myopathy, rash, peripheral neuropathy, insomnia, symptoms related to sleep or concentration, and the most interesting side effects associated with statins. It is known that there is a risk of causing symptoms such as liver toxicity, myopathy, indigestion, abdominal pain. This is a problem in the treatment of hyperlipidemia, which should be managed by steadily administering drugs in the long term.

Korean Patent Laid-Open Publication No. 2004-0066344 discloses an extracellular polysaccharide extract produced from a mycelium liquid culture of a roe deer fungus having an hypolipidemic effect. However, since the extracellular polysaccharide extract disclosed in this patent uses a liquid medium and requires several separation or purification processes, the manufacturing process is not only cumbersome and complicated, but also has a medicinal effect to replace a conventional chemical synthesis drug. It wasn't practically useful.

Therefore, there is a need for the development of a new drug that can significantly reduce the possibility of toxicity or side effects when taken for a long time, and has an excellent effect on the prevention and treatment of hyperlipidemia and cardiovascular disease compared to existing drugs, and can be mass-produced in a simple manner. .

The present invention is to solve the problems of the prior art as described above, while significantly lowering the possibility of toxicity or side effects when taken for a long time, and has an excellent effect in the prevention and treatment of hyperlipidemia and cardiovascular disease compared to conventional drugs, It is a technical task to provide a pharmaceutical composition for the prevention or treatment of hyperlipidemia or cardiovascular disease containing natural extracts which can be mass-produced in a simple manner, and the health functional food for the prevention or improvement of hyperlipidemia or cardiovascular disease.

In order to solve the above technical problem, the present invention is a pharmaceutical for the prevention or treatment of hyperlipidemia or cardiovascular disease containing a solvent extract of a mixture of Roeden mushroom fungus mycelium cultured in Ingeria mugwort medium and the Ingeria mugwort medium used in the culture as an active ingredient. To provide a composition.

According to another aspect of the present invention, there is provided a health functional food for the prevention or improvement of hyperlipidemia or cardiovascular disease containing a solvent extract of the wormwood mycelium mycelium cultured in the ingeria mugwort medium and the mixture of the intake mugwort medium used in the culture as an active ingredient do.

According to the present invention, it can be prepared by a simple method, exhibits a remarkably superior effect to the conventionally used drugs in the prevention or treatment of hyperlipidemia or cardiovascular disease, and the problem of side effects caused by long-term administration of existing synthetic drugs Because it contains a natural extract that can be solved as an active ingredient, it is possible to obtain a pharmaceutical composition and health functional food which is very useful for the prevention or treatment of hyperlipidemia or cardiovascular disease. In particular, the extracellular polysaccharide as disclosed in Korean Patent Laid-Open No. 2004-0066344 shows an effect at the level of 200 mg / Kg / Day, but the extract (HEAC) according to the present invention is administered at only 100 mg / Kg / Day, which is only half thereof. The amount can also give a better effect.

The present invention is based on the new finding that the extract of Roebencus mycelium mycelium cultured in Injinguru medium has an excellent effect of reducing the blood fat content, and the prophylaxis or treatment of hyperlipidemia or cardiovascular disease It relates to a new use. Hereinafter, the present invention will be described in more detail.

<Rose Mushroom>

Roe deer mushroom (Hericium erinaceum) belongs to Aphyllophorales bearded mushroom (Hydnaceae), and is known to have anti-cancer, anti-oxidant, nourishing tonic, immune regulation and biofunction functions. It is used as a raw material for nutraceuticals. In the present invention, the mycelium of roe deer mushroom commonly used in health functional food, etc. can be used without particular limitation.

<Injin mugwort badge>

Injin mugwort (Artermisia capillaris) in Korea is called as iron, mugwort, wormwood, etc., and is commonly known to have a beneficial effect on the liver is used as a raw material of health functional food. In the medium for culturing the Roe deer fungus mycelium in the present invention can be used without any particular limitation to the jinjinjin usually used in health foods.

In the present invention, the term "injin mugwort medium" refers to all mediums including injin mugwort, which may be used as in itself, in addition to rice bran, and may further include water, if necessary. It is not particularly limited. According to one embodiment of the present invention, the Injin mugwort medium preferably contains 10 to 50% by weight, more preferably 15 to 30% by weight, based on the total medium weight of rice bran in addition to the indium mugwort, in which case the extraction yield and activity of the extract An advantageous effect can be obtained in the improvement.

<Cultivation of mycelium>

In the present invention, there is no particular limitation on the method of culturing the Roe worm fungus mycelium on the wormwood medium, for example, mushroom mycelium known by conventional literature such as Korean Patent Application No. 10-2007-0101831, which is incorporated herein by reference. The method of inoculating and incubating in a solid medium can be used as it is or by modifying it appropriately.

The scab mushroom mycelium to be inoculated in the medium is, for example, YMPG (yeast extract malt extract-peptone-glucose, KH ₂ PO ₄ 2.0 g / L, MgSO ₄ .7H ₂ O 1.0 g / L, glucose 10.0 g / L, thiamine- In mushroom mycelium culture medium such as HCl 1.0g / L, DL-asparagine 1.0g / L, peptone 2.0g / L, malt extract 10.0g / L, yeast extract 2.0g / L, agar 20.0g / L) medium After incubation, it may be inoculated in the wormwood medium.

Although not limited thereto, the amount of Rupper fungus mycelium inoculated on the Rhizome medium in this culture step is 5 to 20% by weight, preferably 7 to 15% by weight based on the weight of the medium, and the culture temperature is 20 to 30 ° C, Preferably from 23 to 26 ℃, the incubation time 40 to 50 days may be advantageous in terms of economic efficiency, yield and biological activity of the extract.

The solid medium in which the cultured mushroom mycelium mycelium was formed was used as such or dried to be used in a subsequent extraction step.

According to one embodiment of the present invention, the dried Root wormwood, water and rice bran mixed in sterilized at 110 ℃ to 130 ℃ sterilized at 110 ℃ to 130 ℃, the wormwood mycelium precultured in YMPG medium 10 weight of the weight of the medium Cultivated mycelium at 23 ° C. to 26 ° C. for about 40-50 days after inoculation by%, and drying the solid medium in which the rhinobacteria mushroom mycelium was formed before the fruiting body formation obtained by the cultivation. Water ( Hericium erinaceum Hypha cultivated with Artemisia capillaris , HEAC) can be obtained, which is then used for subsequent extraction.

<Extraction of mycelium>

In the present invention, there is no particular limitation on the method for solvent extraction of the culture cultured on the Root wormwood mycelium fungus mycelium medium, for example, conventional documents such as Korean Patent Application No. 10-2007-0101831, which is incorporated herein by reference. The extraction method known by the above can be used as it is or as appropriately modified.

The extraction solvent may be selected from the group consisting of water, alcohol having 1 to 5 carbon atoms (eg, ethanol), hexaform, chloroform, methylene chloride, acetonitrile, acetone, ethyl acetate, and mixtures thereof. It is not necessarily limited.

Although not limited thereto, the amount of the extraction solvent is preferably 2 to 20 times (volume / weight), more preferably 5 to 15 times (volume / weight) in the volume ratio per weight of the culture to be extracted. If the amount of the extraction solvent is too small than the above-mentioned range, the extraction efficiency may be lowered. If the amount of the extraction solvent is too high, the cost and time required for post-treatment such as filtration / concentration increase are not preferable. According to a preferred embodiment of the present invention, it is very suitable in terms of economics and extraction yield to extract 80% ethanol using 10 times (volume / weight) in the volume ratio per weight of the culture to be extracted.

There is no limit to the number of extractions. Therefore, one extraction may be sufficient, and one or more of the above solvents may be added to the extract once, and may be reextracted two or more times. Extraction may be performed by methods commonly used in the preparation of plant extracts, such as stirring extraction, hot water extraction, and the like. The extraction temperature and time may be appropriately selected depending on the amount of the object and / or the extraction solvent and the working conditions, for example, extraction may be performed at 40 to 60 ° C. for 12 to 48 hours.

The extraction result can be filtered and / or concentrated by conventional workup methods. Filtration and / or concentration steps may also be carried out by conventional extract workup methods, for example at 20 to 60 ° C., preferably at 30 to 50 ° C.

According to one embodiment of the present invention, 80% (v / v) ethanol was added 10 times (volume / weight) to the dried product of the culture cultured in the wormwood mycelium medium, and then extracted at 40 ℃ for 24 hours, Filtration and concentration give HEAC ethanol extract.

The extract obtained as described above is preferably 0.05 to 0.15% by weight, more preferably 0.08 to 0.1% by weight of ethyl palmitate as an active ingredient identified by GC; 0.1-0.2 wt% of 1-octanol (1-Octanol), more preferably 0.12-0.15 wt%; 0.05 to 0.15% by weight of ethyl oleate, more preferably 0.09 to 0.12% by weight; 0.05 to 0.15% by weight of ethyl stearate, more preferably 0.09 to 0.12% by weight; Phytol 0.05 to 0.15% by weight, more preferably 0.08 to 0.1% by weight (UV quantification); 0.05 to 0.15% by weight of ethyl linoleate, more preferably 0.08 to 0.12% by weight; 0.05 to 0.15% by weight of ethyl linolenate, more preferably 0.08 to 0.12% by weight; 0.05 to 0.12% by weight of ethyl eicosapentanoate, more preferably 0.07 to 0.1% by weight; Eicosanol 0.05 to 0.15% by weight, more preferably 0.07 to 0.1% by weight; And 6,7-dimethoxycoumarin (6,7-dimethoxycoumarin, also known as 'scoparone') from 0.04 to 0.12 wt%, more preferably from 0.05 to 0.1 wt% (HPLC quantitative).

Among the active ingredients identified above, 6,7-dimethoxycoumarin and ethyl linoleate are open to blood fat reduction effect. However, these compounds are present in traces (1% or less) in the HPLC analysis results of the extracts of the Roeden mushroom mycelium cultures cultured in the Insamui wormwood medium of the present invention, and the extract of the present invention is better than the case in which each of them is used alone in the same amount ( That is, in the case of the component is used in an amount of 100 times or more) excellent hyperlipidemia and cardiovascular disease treatment effect (see Experimental Example here), this excellent effect represented by the extract of the present invention is 6,7-dimethoxycoumarin From the nature of the natural extracts, rather than only by ethyl linoleate itself, it can be interpreted as a synergistic effect between the other active ingredients in the extract or these components.

<Pharmaceutical composition>

The pharmaceutical composition according to the present invention contains a solvent extract of a mixture of Roeden mushroom mycelium cultured in Ingeria mugwort medium obtained from the above, and a mixture of Ingeria mugwort medium used for cultivation thereof as an active ingredient, and is used for hyperlipidemia or cardiovascular disease. It has a prophylactic or therapeutic effect.

In view of the fact that the active ingredient extract of the present invention is a natural extract, the pharmaceutical composition according to the present invention is expected to have less concern of side effects during long-term in vivo administration than other synthetic pharmaceuticals.

The term "cardiovascular disease" as used herein more specifically includes, but is not limited to, diseases such as arteriosclerosis, angina pectoris, myocardial infarction, and hypertension.

As used herein, the term “prevention” is a term used to mean prophylactically administering an agent, such as a pharmaceutical composition, to a healthy patient to prevent the onset of the diseases and symptoms referred to herein. It is also meant to include prophylactic administration of such agents to patients who are in the pre-stage of the disease to be treated.

As used herein, the term “treatment” refers not only to the treatment of the diseases referred to herein, but also to the prophylactic administration of a combination formulation, such as a combination or pharmaceutical composition, to healthy patients for the development of the diseases and symptoms referred to herein. To prevent.

The composition of the present invention, in addition to the solvent extract of the mixture of the Roe mushroom fungus mycelium cultured in the Root Artemisia medium and the Root Artemisia medium used for the cultivation of the present invention, auxiliaries such as pharmaceutically suitable and physiologically acceptable carriers, excipients and diluents It may contain. As used herein, a "pharmaceutically acceptable carrier" is a term that refers to a composition, specifically an ingredient other than the active substance of the pharmaceutical composition.

Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants can be used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like can be mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, transdermal agents and the like. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The dosage of the composition can be appropriately adjusted according to the condition, age, weight, degree of disease, route of administration, etc. of the patient, and may be administered once a day at regular time intervals or several times a day at regular time intervals according to the judgment of a doctor or pharmacist. It can also be administered in divided doses. For example, the dosage is based on the active ingredient content, 0.0001 mg / kg / day to 1000 mg / kg / day, preferably 0.01 mg / kg to 500 mg / kg, more preferably 0.01 mg / kg to 100 mg / kg may be, but is not limited thereto. The above dosages are illustrative of the average case and may be high or low depending on individual differences. If the daily dose of the composition of the present invention is less than the dose, a significant effect may not be obtained, and if it exceeds the above, it may be uneconomical and out of the range of normal dose, which may cause undesirable side effects. It is good to set it as the said range.

The composition according to the invention can be administered to a mammal, including humans, by various routes. The mode of administration can be any of the routinely used forms and can be administered, for example, by oral, rectal or intravenous, intramuscular, or subcutaneous injection. The compositions of the present invention may be formulated in oral dosage forms, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, or parenteral formulations in the form of transdermal, suppository, and sterile injectable solutions according to conventional methods. Can be used.

In an embodiment in which the composition of the present invention is applied to a human, the composition of the present invention may be administered alone, but is generally mixed with a pharmaceutical carrier selected in consideration of the mode of administration and standard phamaceutical practice. May be administered. For example, the compositions of the present invention may be in the form of tablets containing starch or lactose, in the form of capsules alone or in the form of excipients, or in the form of elixirs or suspending agents containing chemicals to flavor or color. It can be administered orally, orally or sublingually. Such liquid preparations may be suspending agents (e.g., semisynthetic glycerides such as methylcellulose, witepsol or mixtures of apricot kernel oil and PEG-6 esters or PEG-8 and caprylic / capric Pharmaceutically acceptable additives such as glyceride mixtures such as mixtures of glycerides).

<Health functional food>

The health functional food according to the present invention contains a solvent extract of a mixture of Roeden mushroom fungus mycelium cultured in the Root wormwood medium obtained in the above manner and the Root wormwood medium used for cultivation thereof as an active ingredient, and the hyperlipidemia or cardiovascular disease It has a preventive or improving effect.

Health functional food according to the present invention may be a variety of food, beverages, food additives and the like. The content of the extract as an active ingredient contained in the health functional food may be appropriately, for example, 0.01 to 15% by weight of the total food weight, without particular limitation depending on the form of the food, the desired use, etc. It may be a ratio of 0.02 to 10 g, preferably 0.3 to 1 g based on 100 ml.

The health functional food of the present invention may be prepared and processed in a form including, but not limited to, tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of preventing or improving hyperlipidemia or cardiovascular disease. The term "health functional food" used in the present invention refers to a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Act No. 6727 of the Health Functional Food Act. It means the ingestion for the purpose of obtaining a beneficial effect for health use such as nutrient control or physiological action.

The health functional food of the present invention may include a conventional food additive, and the suitability as the "food additive" is applicable in accordance with the General Regulations of the Food Additives Code and General Test Methods, etc. approved by the Food and Drug Administration, unless otherwise specified. Judge according to the standards and standards for the item. Examples of the items listed in the "Food Additive Revolution" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid, natural additives such as color pigments, licorice extracts, crystalline cellulose, high color pigments, guar gum, And mixed preparations such as sodium L-glutamate preparation, noodle addition, preservative preparation, and tar coloring preparation.

The health functional food of the present invention may include additives such as suitable carriers, excipients, diluents, and the like used in conventional food production in conventional contents. For example, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protection Sex colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages and the like. These ingredients may be used independently or in combination, and the amount thereof is not particularly limited, but is generally selected from 0 to 20 parts by weight per 100 parts by weight of the active ingredient extract.

For example, a dietary supplement in the form of tablets may be obtained by granulating a mixture of excipients, binders, disintegrants, and other additives in a conventional manner, and then compressing the mixture with a lubricant or the like. Can be directly compression molded. In addition, the health functional food in the form of tablets may contain a mating agent and the like, if necessary, may be coated with a suitable coating agent.

Hard capsules of the health functional foods in the form of capsules may be prepared by filling an extract, which is an active ingredient, in a conventional hard capsule with a mixture of additives such as excipients, or granular or peeled granules thereof, and the soft capsule is an active ingredient. The extract may be prepared by filling a mixture with an additive such as an excipient into 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 cyclic health functional food can be prepared by molding a mixture of an active ingredient extract, excipient, binder, disintegrant, etc. by a suitable method. You can also be blessed with a substance.

The health functional food in the form of granules may be prepared by granulating a mixture of an extract, an excipient, a binder, a disintegrant, and the like as an active ingredient in a suitable manner, and may contain a flavoring agent, a copper, and the like as necessary.

The term definitions of the excipients, binders, disintegrants, glidants, copulation agents, flavoring agents, etc. of the present invention are described in documents known in the art and include those having the same or similar functions. Sacrament, Korean College of Pharmacy, 5 revised edition, p33-48, 1989).

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

[Example]

1. Preparation Example of Extract

1 kg of dried ginseng mugwort and 2 L of water (including 20% by weight of rice bran) were mixed and sterilized at 110 ° C. to 130 ° C. for about 3 hours to prepare a ginseng solid medium. YMPG (yeast extract malt extract-peptone-glucose, KH ₂ PO ₄ 2.0g / L, MgSO ₄ · 7H ₂ O 1.0g / L, glucose 10.0g / L, thiamine-HCl 1.0g / L, DL-aspa 1.0 g / L of Razine, 2.0 g / L of Peptone, 10.0 g / L of Malt Extract, Yeast Extract 2.0 g / L, 20.0 g / L of Agar) Mycelia were incubated for about 40-50 days at 23 ℃ to 26 ℃.

The solid medium in which the rotiferous mycelium mycelium was formed before the fruiting body formation obtained by the cultivation was dried to obtain a Hericium erinaceum Hypha cultivated with Artemisia capillaries (HEAC).

400 g of 80% (v / v) ethanol was added to 40 g of the HEAC dried product, which was extracted at 40 ° C. for 24 hours, filtered, and concentrated to obtain 4.4 g of HEAC ethanol extract (yield: 11.0%).

Gas chromatography (GC) was performed on the HEAC ethanol extract obtained above to obtain a result as shown in FIG. 1.

The GC analysis conditions are as follows:

Column: HP-10, Carrier gas: Helium, Flow rate: 1.5ml / min

Injection volume: 1.0 μl,

Injector temp .: 280 ℃

Oven temp .: start at 80 ℃, stand for 5.0min, increase 20 ℃ / min,

stand for 5.0 min

Termanal auxl temp .: 300 ℃, Detector voltage: 1.3kV

Each peak component and content found by analyzing mass spectrometery (MS) spectra for each of the obtained peaks of FIG. 1 are as follows.

Ethyl palmitate: 0.0925 wt%

② 1-octanol (1-Octanol): 0.135% by weight

③ ethyl oleate: 0.1 wt%

④ ethyl stearate: 0.104 wt%

⑤ Phytol: 0.092% by weight

⑥ Ethyl linoleate: 0.095 wt%

⑦ Ethyl linolenate: 0.0975 wt%

⑧ Ethyl eicosapentanoate: 0.0775 wt%

⑨ Eicosanol: 0.0875% by weight

⑩ 6,7-dimethoxycoumarin (0.07 wt%)

The MS analysis conditions above are as follows:

Column: HP-10, Carrier gas: Helium, Flow rate: 1.5ml / min

Injection volume: 1.0 μl, Injector temp .: 280 ° C

Oven temp .: start at 80 ℃, stand for 5.0min, increase 20 ℃ / min,

stand for 5.0min

Termanal auxl temp .: 300 ℃, Detector voltage: 1.3kV

Library: NIST. shimadzu Co.

In addition, the structure of the 11-minute peak separation material (peak # 7) obtained by performing HPLC on the obtained HEAC ethanol extract was confirmed by GC-MS. As a result, the substance was found to be 6,7-dimethoxycoumarin.

The HPLC analysis conditions are as follows:

Mobile Phase-Acetonitrile: Buffer (400)

(Buffer solution-3.0 mL acetic acid, 6.0 g ammonium acetate, 600.0 mL H ₂ O)

Flow rate-0.5 mL / min

UV Absorbance-360 nm

Column-Capcell pak C ₁₈ (4.6 mm × 250 mm, 5 μm)

Injection volume-20.0 μL

Sample solution-70% (v / v) ethanol

Sample concentration-sample: solution = 20.0 mg: 20.0 mL

The analyzed components are as follows.

2. Experimental Example of Pharmacological Effects

Induction of hyperlipidemia using high fat diet in white rats, after administration of HEAC ethanol extract and control agent alone or in combination, serum was separated and serum total cholesterol, triglyceride, HDL- Changes in arteriosclerosis risk index, which is a risk index of cholesterol, LDL-cholesterol, glutamic oxaloacetic transaminase (GOT) index, glutamic pyrovic transaminase (GPT) index, and cardiovascular disease, were analyzed. The details are as follows.

2.1. Experimental Materials and Methods

2.1.1. Set up experimental animals and normal, control and experimental groups

As the experimental animals, male 7-week-old SPF Sprague-Dawley rats (210 ± 10 g in weight, orient bio) weighing 210 ± 10 g were used (eight in each group). The general diet used AIN-93G type (pellet form, Feedlab Korea), and the high fat diet for hyperlipidemia was used High Fat Diet 45Kcal% type (pellet form, Feedlab Korea). The general diet and high fat diet are summarized in Table 1 below.

Normal, control, and experimental groups were prepared as follows (n = 8 for each group):

1) Normal group: A group of white rat males who were raised on a regular diet without inducing hyperlipidemia and did not receive therapeutic drugs.

2) Negative Control Group-1: A group analyzed by serum analysis after completion of hyperlipidemia by high fat diet.

3) Negative Control Group-2: A group fed with a high-fat diet, induced hyperlipidemia, without administration of a therapeutic drug and until the completion of the treatment period.

4) positive control group

Induced hyperlipidemia and then administered with each of the following substances (dose: 100 mg / kg):

Simvastatin: Coronary artery disease, hyperlipidemia (40mg as Simvastatin, product name: Simbarod tablet, Chong Kun Dang)

Atorvastatin: Coronary artery disease, hyperlipidemia (40 mg as Atorvastatin, product name: Lipirow tablet, Chong Kun Dang)

-Ethanol extracts of the scaber mycelium mycelium alone: 80% (v / v) ethanol extracted at 40 ℃

-Ethanol extract of Insam wormwood alone: extracted at 40 ° C with 80% (v / v) ethanol

6,7-dimethoxycoumarin (Scoparone) alone (Sigma-Aldrich, 98%)

Ethyl linoleate alone (Sigma-Aldrich, at least 99%)

5) Experiment group

Induction of hyperlipidemia, the group administered with 200 mg / kg, 100 mg / kg and 50 mg / kg of the HEAC ethanol extract obtained in the preparation example, respectively

The normal, control, and experimental groups are summarized in Table 2 below:

2.1.2. Induction of Hyperlipidemia

After 1 week of adaptive breeding with the general diet before the start of the experiment, the normal group raised the diet and purified water for 4 weeks (28 days), and the control group and the experimental group fed the high fat diet and purified water for 4 weeks ( 28 days) to induce hyperlipidemia. Daily dietary intake, dietary efficiency, and weight change were measured.

2.1.3. Treatment after hyperlipidemia

After 4 weeks of hyperlipidemia induction, control-1 was immediately dissected for 16 hours and serum was analyzed to investigate the level of physiological activity after lesion induction was completed.

The normal group was fed for 2 weeks (14 days) by autonomous feeding of normal diet and purified water. Negative control group-2 was further fed for 2 weeks (14 days) by oral administration (3 times a day) of 3 mL of saline solution without administration of any treatment drug, while receiving the autonomic diet and purified water after completion of hyperlipidemia. ). After completion of hyperlipidemia, the positive control group and the experimental group autonomously fed the diet and purified water, and then raised orally (once a day) each of the therapeutic drugs for two weeks (14 days) to raise them (treatment group). The therapeutic drug was suspended orally in 3 mL of saline solution according to the dose. Their daily dietary intake, dietary efficiency and weight change were measured.

2.1.4. Blood collection and serum separation

1) Sampling of Negative Control-1

After the hyperlipidemia induction was completed, eight experimental animals of negative control group-1 fasted for 16 hours were killed by diethyl ether anesthesia, and then opened along the median of the abdomen and collected through the aorta. Blood was left at 25 ° C. for 30 minutes, centrifuged at 3000 rpm for 20 minutes, and serum obtained through centrifugation was used for hemochemical analysis.

2) Sampling of normal group, negative control group-2, positive control group and experimental group

After the induction and treatment of hyperlipidemia was completed, the experimental animals of each group were fasted for 16 hours, and then serum obtained using the same method as the sampling method of the negative control group-1 was used for hemochemical analysis. Serum total cholesterol, triglyceride, HDL-cholesterol, GOT / GPT kit kit solution used in this analysis was used from Asan Pharmaceutical.

2.1.5. Biochemical Analysis of Serum

1) Determination of Total Cholesterol (Active Levels related to Fatty Liver and Hyperlipidemia), Triglycerides (Active Levels Related to Fatty Liver and Hyperlipidemia), HDL-Cholesterol (Activity Levels Related to Atherosclerosis Prevention):

It was measured using kit solution (Asan Pharmaceuticals) prepared by enzymatic method and expressed in mg per dL of serum.

2) Determination of LDL-cholesterol (activity level related to atherosclerosis):

It was calculated by the Friedwald method of the following formula and expressed in mg per dL of serum.

LDL-cholesterol = total cholesterol-HDL cholesterol- (triglyceride / 5)

3) Measurement of Atherosclerosis Risk Index (AI):

It calculated by the Haguland method of the following formula.

Atherosclerosis risk index (AI) = (total cholesterol-HDL cholesterol) / HDL cholesterol

4) Determination of GOT / GPT index (fatty liver, liver damage, general liver activity indicator):

Prepared by Reitman-Frankel's method (Reitman S., Frankel SA: Colorimetric method for the determination of serum glutamic oxaloacetic acid and glutamic pyruvic transaminases, Am. J. Clin. Patrol., 1957; (28): 58-63) Measured using a kit (Asan Pharmaceuticals), 1 unit per ml of serum (unit: 1 ml of serum at 32 ° C, 1 min when absorbance decreased by 0.001 at 340 nm in 1 min, reference: same as Reitman-Frankel method) ).

2.1.6. Statistical processing of results

The experimental results obtained by the biochemical analysis of the serum are expressed as mean and standard deviation. In addition, the significance test of each population was carried out by Student's t-test (P <0.005).

2.2. result

The total cholesterol, triglycerides, HDL-cholesterol and LDL-cholesterol obtained above are shown in Table 3 below, and the atherosclerotic risk index, GOT index, and GPT index, respectively, in Table 4 below.

As can be seen from the above experimental results, the ethanol extract of the mixture of the Roe mushroom fungus mycelium cultured in the Roots of the Roots of the present invention and the Roots of the Roots used in the culture is simvastatin, a conventional drug in the treatment and prevention of hyperlipidemia or cardiovascular disease And HDL-cholesterol, an active level related to cardiovascular disease prevention, is much higher than that of atorvastatin as well as known active ingredients 6,7-dimethoxycoumarin and ethyl linoleate alone in the same amount. In addition, the arterial risk index was much lower (good effect), and the synergistic effect was also significantly superior to that of the extract of Rhizome mycelia and Rhizome alone.

1 is a gas chromatographic result spectrum of the HEAC ethanol extract prepared in the embodiment of the present invention.

<Explanation of symbols in the drawings>

[Horizontal axis: time (minutes), vertical axis: relative peak intensity (arbitrary units)]

① ethyl palmitate: 5.7 minutes

② 1-octanol: 6.2 minutes

③ ethyl oleate: 6.5 minutes

④ ethyl stearate: 8.2 min

⑤ Pytol: 9.3 minutes

⑥ ethyl linoleate: 9.9 minutes

⑦ ethyl linoleate: 10.2 minutes

⑧ Ethyl Eicosaptanoate: 11.63 minutes

⑨ Eicosanol: 13.6 minutes

⑩ 6,7-dimethoxycoumarin: 8.8 minutes

Claims (12)

A pharmaceutical composition for the prevention or treatment of hyperlipidemia or cardiovascular disease, which comprises a solvent extract of a mixture of Roeden mushroom mycelium cultured in Ingeria mugwort medium and the mixture of Ingeria mugwort medium used in the culture thereof as an active ingredient. 2. The pharmaceutical composition of claim 1, wherein the erythropoiete medium further comprises rice bran. The pharmaceutical composition according to claim 1, wherein the solvent for extraction is selected from the group consisting of water, alcohols having 1 to 5 carbon atoms, hexane, chloroform, methylene chloride, acetonitrile, acetone, ethyl acetate, and mixtures thereof. . The pharmaceutical composition of claim 1, wherein the extraction solvent is ethanol. The method of claim 1, wherein the extract is ethyl palmitate, 1-octanol, ethyl oleate, ethyl stearate, phytol, ethyl linoleate, ethyl linoleate, ethyl eicosaptanoate, eicosanol and 6, A pharmaceutical composition comprising 7-dimethoxycoumarin. The pharmaceutical composition of claim 1, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, angina pectoris, myocardial infarction, and hypertension. A health functional food for the prevention or improvement of hyperlipidemia or cardiovascular disease, which contains a solvent extract of a mixture of Roeden beet mushroom mycelium cultured in Ingeria mugwort medium and the Ingeria mugwort medium used for cultivation thereof as an active ingredient. 8. The dietary supplement of claim 7, wherein the Injin mugwort medium further comprises rice bran. The health functional food according to claim 7, wherein the extraction solvent is selected from the group consisting of water, alcohols having 1 to 5 carbon atoms, hexane, chloroform, methylene chloride, acetonitrile, acetone, ethyl acetate, and mixtures thereof. . The health functional food according to claim 7, wherein the extraction solvent is ethanol. The method of claim 7, wherein the extract is ethyl palmitate, 1-octanol, ethyl oleate, ethyl stearate, phytol, ethyl linoleate, ethyl linoleate, ethyl eicosaptanoate, eicosanol and 6, A dietary supplement comprising 7-dimethoxycoumarin. The health functional food according to claim 7, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, angina pectoris, myocardial infarction, and hypertension.

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