KR100829691B1 - Composition for promoting degradation of lipid containing plant extracts - Google Patents

Abstract

Composition for promoting lipid degradation according to the present invention by inducing the activity of the IDH3α gene promoter by containing one or more extracts selected from the group consisting of liver extract, rake extract, silk extract, Ulleung chrysanthemum extract and Paddungmul extract as an active ingredient It activates the TCA circuit and thereby promotes the oxidative degradation of lipids, and can further be used for the treatment or prevention of obesity, the treatment or prevention of hyperlipidemia.

Obesity, IDH3α, TCA Circuits, Rake, Yeast, Ulleung Chrysanthemum

Description

Composition for promoting degradation of lipid containing plant extracts

1 is treated with 1.25, 2.5, 5, and 10 ppm of liver extract, rake extract, yeast extract, Ulleung chrysanthemum extract, pine nut extract, and paddock sprout extract to Huh7 cells, a hepatocyte cell line containing IDH3α promoter and luciferase fusion construct It shows luciferase activity according to the concentration at the time of doing.

Figure 2 shows the effect of reducing total cholesterol by the pine leaf extract and bark extract.

Figure 3 shows the LDL cholesterol reduction effect by the pine leaf extract and bark extract.

The process of breaking down lipids and using them as energy sources can be divided into three stages. In the first stage, these are decomposed into constituents such as fatty acids, and the second stage is oxidative (-oxidation), and then acetyl-CoA (acetyl-CoA, hereinafter referred to as acetyl-CoA). in the tricarboxylic acid cycle. Various energy sources are metabolized into acetyl-CoA, which is oxidized to carbon (CO ₂ ) and water (H ₂ O) (TCA cycle), all of which are enzymes present in the mitochondria matrix or in the inner membrane. Caused by the military The TCA cycle is a ring-shaped metabolic process, also called the citric acid cycle (citrate cycle or Krebs cycle), and the reaction first involves a four-carbon compound, oxaloacetate, and an acetyl-CoA with two carbon atoms. Is combined to produce six carbon atoms citric acid having three carboxyl groups (-COOH). Citric acid is converted to an isocitrate by shifting the position of a hydroxyl group (-OH) by an aconitase enzyme, which is an isocitrate dehydrogenase (“IDH3α”). Oxalosuccinate, an intermediate in the state bound to the enzyme, reduced coenzyme NAD +, and is immediately decarbonized to produce alpha-ketoglutarate and carbon (CO ₂ ). Create The generated NADH is immediately transferred to the electron transport system present in the mitochondrial substrate to generate ATP, which is an active energy in vivo. Therefore, if IDH3α activity is induced, the activity of the TCA circuit is enhanced, and thus lipids can be finally oxidized and decomposed to generate ATP energy (Cupp JR, MaAlister Henn L., Journal of Biological Chemistry. 22199-22205 (1991). )). On the other hand, when the TCA circuit has a problem, it is known that ATP synthesis, which is an active energy in vivo, decreases, fatty acid synthesis increases, oxidation also decreases, and eventually obesity is formed (Wlodek D, Gozales M. , Journal of Theoretical Biology., 33-44 (2003)).

The present invention relates to herbal and native plant extracts that increase the activity of the TCA cycle in the liver, and more particularly to increase the activity of a gene promoter called IDH3α, a protein that plays an important role in the activity of the TCA cycle in human liver cell lines. In the end, it is related to the extract of native plants, such as rakes, yeasts, Ulleung chrysanthemums, pine, wild herbs and herbal extracts, which play a role in activating the TCA circuit.

Accordingly, the present inventors searched for hundreds of herbal and native plant extracts to activate plant promoters that activate the IDH3α promoter in hepatocytes, thereby activating the TCA circuit and promoting oxidative degradation of lipids. We found the efficacy of Ulleung chrysanthemum, pine, parsnip and herbal extract.

Accordingly, it is an object of the present invention to provide a composition that activates the TCA cycle in the liver to promote complete oxidation and degradation of lipids. Furthermore, the present invention provides a composition containing a plant extract as an active ingredient effective for the prevention or treatment of obesity, the prevention or treatment of hyperlipidemia through the promotion of complete oxidation and degradation of lipids.

In order to achieve the above object, the composition for preventing or treating obesity according to the present invention contains one or more extracts selected from the group consisting of liver extract, rake extract, yeast extract, Ulleung chrysanthemum extract and Paddungmul extract as active ingredients. Characterized in that.

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In addition, the composition for preventing or treating hyperlipidemia according to the present invention is characterized in that it contains one or more extracts selected from the group consisting of soybean extract, rake extract, yeast extract, Ulleung chrysanthemum extract and Paddungmul extract as an active ingredient.

In the composition according to the present invention described above, the composition preferably activates the TCA circuit in the liver.

In the composition according to the present invention described above, the composition preferably increases the activity of the isocitrate dehydrogenase enzyme (IDH3α) gene promoter.

In the above composition according to the present invention, the extract is not particularly limited but is preferably an extract of ethanol or methanol.

Most preferably the ethanol is 95% ethanol.

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

Belamcandae Rhizoma ( Belamcanda chinensis Leman. ) Is a rhizome herb of panax irises (Iris and Iridaceae), divided into uneven chunks, 3-10 cm long and 1-2 cm in diameter. The outer side is yellowish brown with crumpled and dense patterns. There are some traces of several stems that have fallen off, and some have remained. The lower part has thin roots or traces of roots. The quality of this drug is hard and the cut side is yellow. This medicine has no smell and tastes bitter and spicy. The main ingredients include belamcandin, iridin, tectoridin, tectorigenin, magniferink and isotectoridine. As pharmacological action, anti-inflammatory action, diuretic action, expectorant action, antifungal action is mentioned.

Galium spurium is a two-year-old vine plant of the genus Magnolia of the dicotyledonous plant. It lives in Korea, Japan, Sakhalin, etc., and its main stem is 60 ~ 90cm long and has thorn hairs facing down to each ridge. Stick. In spring, young sprouts are eaten as herbs, and herbs are harvested from July to September, and dried soybeans are called mountain peas, and bruises, pain, neuralgia, gonorrhea, hematuria, enteritis, boils, and carcinomas. Used to treat the back. The rake extract contained in the composition according to the present invention preferably uses outpost.

Persicaria hydropiper , also known as a medicinal plant, a medicinal plant, and a medicinal plant, grows in wetlands or streams, grows from 40 to 80 cm in height, has no hairs, and branches off. The leaves are spicy, and in Japan they are used for fish dishes that are sprouting. It has a continuous hemostatic effect and is used for uterine bleeding, hemorrhoidal bleeding and other internal bleeding. Leaves and stems have excellent antibacterial action, lower blood pressure and strengthen the tension of the small intestine and uterus. Civilians crush this and use it to catch fish. In the composition according to the present invention, the extract is preferably used outpost.

Ulleung Chrysanthemum ( Chrysanthemum zawadskii var. Lucidum ) is a perennial herb of the dicotyledon plant Lantern, a native species of Korea, distributed in Ulleungdo. It is a kind of wild chrysanthemum, grows in the mountains of the seashore, and is about 30cm high. Flowers are used for ornamental purposes, and in oriental medicine, the whole abandonment is used as medicinal herbs such as gynecological diseases, stroke, anorexia and neuralgia. In the composition according to the present invention, the Ulleung chrysanthemum extract is preferably used outpost.

Pine tree ( Pinus koraiensis ) is an evergreen tree of the pine tree family, the soybean plant, and is also called Hongsong. It grows above 1,000m above sea level, and is a large tree with a height of 20-30m and a diameter of 1m. Seed without wings, ripens in October of next year, 12-18mm long, 12mm diameter, edible or medicinal. The belly milk contains 74% fat oil and 15% protein and has a nutritious tonic effect. In the composition according to the present invention, it is preferable to use a stem-bark of the pine tree extract.

Cryptotaenia japonica is a perennial herb of the dicotyledonous cultivars Asteraceae , also called bandinae . It grows in mountainous areas and is 30 ~ 60cm high. The stem is straight and slightly cracked, and there is no hair and fragrance all over. Stems and leaves are green with roots thick. Leaves are alternate, petioles are 5 ~ 17cm long and consist of three small leaves with glossy on the back. Young leaves are edible and grown as vegetables. What is collected in summer is dried in the shade and used as a medicine for goiter. In the composition according to the present invention, it is preferable to use an outpost for the extract.

In the composition according to the present invention, ethanol, methanol, butanol, ethyl acetate, chloroform or hexane may be used as a solvent for preparing each extract. Among them, 95% ethanol or methanol is most preferred.

The composition according to the present invention may be a pharmaceutical composition or health functional food composition.

Each extract of the present invention can be prepared by the following method. First, each medicine or plant is dried for 3 to 6 days, preferably 5 days in the shade at room temperature, and then 40 to 60 ° C using ethanol, preferably 95% ethanol or methanol, butanol, ethyl acetate, chloroform, or nucleic acid. Preferably it is extracted for 24 to 48 hours at 50 ℃ and concentrated under reduced pressure to 40 to 50 ℃, preferably 45 ℃. Then, in order to remove traces of solvents that have not been removed in the extraction and concentration process, a small amount of distilled water is added to each active fraction content to homogeneously suspend and freeze-dried to obtain each active fraction in powder form. have. It is preferable to use 95% ethanol as the extraction solvent in the case of liver extract, which is a medicinal herb, and methanol extract in the case of extracts of native plants such as rakes, yeast, Ulleung chrysanthemums, pine trees, and wild herbs.

Each of the extracts according to the present invention may be prepared and used directly according to the preparation method described above, but may also be obtained according to various methods commonly known in the art. In addition, each extract according to the present invention can be easily obtained as a product of a freeze-dried powder state. For example, it can be used to receive each of the extracts from the Korea Plant Extract Bank in Daejeon, South Korea.

The present inventors measured the activity of the IDH3α promoter by measuring the activity of luciferase by treating the Huh7 cell line containing the IDH3α-luciferase fusion gene with each of the extracts in order to determine the effect of promoting the IDH3α promoter activity of each extract. , Luciferase activity was confirmed to increase the effect.

The composition according to the present invention preferably contains any one or more of the extract as an active ingredient in an amount of 0.0001 to 99.9% by weight based on the total weight of the total composition. More preferably 0.1 to 60.0% by weight, considering effective effects and stability, and formulation stability.

The pharmaceutical composition according to the present invention may be administered orally, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous.

In addition, the dosage of the active ingredient will vary depending on the age, sex and weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of those skilled in the art, and generally the dosage ranges from 0.001 mg / kg / day to approximately 2000 mg / kg / day. More preferred dosages are from 0.5 mg / kg / day to 2.5 mg / kg / day.

On the other hand, depending on the intended dosage form, the pharmaceutical composition may be in solid, semisolid or liquid dosage form. Examples of dosage forms include, but are not limited to, tablets, capsules, suppositories, small bags, granules, powders, creams, lotions, ointments, gels, patches, sprays, plasters, liquid solutions, suspensions, dispersions, emulsions, syrups, and the like. . The active ingredient may be encapsulated in liposomes, microparticles or microcapsules.

Solid compositions such as tablets, pills, granules and the like in the formulations may be conveniently coated and typically compositions for intravenous administration are solutions in sterile isotonic aqueous buffer and include local anesthetics to relieve pain at the injection site.

In addition, if desired, the medicament may also contain small amounts of nontoxic auxiliary substances, such as wetting agents, emulsifiers, pH buffers, and the like, examples of which include, but are not limited to, sodium acetate, sorbitan monolaurate, triethanolamine and triethanolamine oleate. It is not limited.

In addition, the pharmaceutical compositions of the present invention may further comprise excipients, carriers and diluents such as stabilizers, antioxidants, binders, colorants, flavors, preservatives and thickeners.

In addition, the present invention provides a dietary supplement composition comprising a food supplement acceptable food additive in addition to the extract. Examples of the food composition include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

At this time, the amount of the extract in the food or beverage can be added in 0.001 to 99.9% by weight of the total food weight, the health beverage composition is based on 100 ml of 0.001 to 0.1 g, more preferably 0.05 to 0.1 g ratio Can be added.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the extract as an essential ingredient in the indicated ratios, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

On the other hand, the extract of the present invention is not so serious toxicity and side effects can be used with confidence even for long-term use for the purpose of prevention.

Hereinafter, the present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1. Analysis of IDH3α Promoter Activity of Plant Extracts

Step 1: Cell Line and Cell Culture

Huh7 (Japanese Collection of Research Bioresources, Cat No JCRB0403), a human hepatoma cell line (Dulbecco's modified Eagle's Medium, Gibco Cat No 1210-0038) containing 10% fetal calf serum Were incubated in 37 ° C., 5% CO ₂ incubator. 1 μg of a fusion vector fused with neomycin (neoselective label) and 3 μg of lipofectamine (Invitrogen Cat No 18324-012) with IDH3α promoter-luciferase for continuous expression of the gene in cells were added to 0.2 ml DMEM. After mixing, the mixture was maintained for 45 minutes. Thereafter, 0.8 ml of DMEM was further added, followed by transformation to Huh 7 cells. Two days later, the cells were replaced with 10% bovine serum DMEM medium containing 0.5 mg / ml G418 (also called neomycin) and cultured for more than one month to obtain cells resistant to G418.

Step 2: increase promoter activity by plant extracts

Sogan extract (preparation number: CA01-094, site: outpost), rake extract (preparation number: 005-001, site: outpost) obtained from Korea Plant Extract Bank (Daejeon, Korea), extract (preparation number: 008) -065, site: outpost), Ulleung chrysanthemum extract (preparation number: 001-166, site: outpost), pine tree extract (preparation number: 015-080, site: stem-bark) and paddock sprout extract (preparation number: 006- 094, site: outpost) using the following experiment.

Each extract of the dry powder state was dissolved in a DMSO (dimethyl sulpholxide) solvent and then treated to the cells prepared in step 1.

The cell lines cultured in step 1 were each treated with 3 ml / flask of trypsin to make a single cell suspension and incubated in 96 well plates at an amount of 1.0 × 10 ⁴ cells / well and incubated for 24 hours. Thereafter, each extract prepared above in DMEM medium was added at a concentration of 1.25, 2.5, 5, 10 ppm, and the cells were treated for 24 hours. After treatment, the activity of the promoter was determined by measuring the activity of luciferase expressed using a Luciferase assay kit (Promega, USA, Cat No # E1501). In order to compare the effects of the extracts according to the present invention, the activity of IDH3α was analyzed in the same manner to the group treated with no treatment (negative control) and the positive control treated with 10 ppm of genistein. The results are shown in FIG. As shown in Figure 1, compared with the negative control (positive control) and positive control (Genystein) it was confirmed that the extracts according to the present invention increased luciferase activity depending on the concentration.

Example 2 Total Cholesterol Inhibitory Effect and LDL Cholesterol Inhibitory Effect by Pine Tree Extract

 Eight weeks-old male ob / ob mouse genetically induced obese bark and leaf extracts were administered for two weeks, and blood was collected to measure total cholesterol and LDL cholesterol in blood. The control group was free to consume AIN93G (American Institute of Nutrition 93G) feed (Reeves, PG (1997) Components of the AIN-93 diets as improvements in the AIN-76A diet.J. Nutr. 127: 838S-841S). The group was free to consume AIN93G feed containing 0.5% of pine bark and leaf extracts. After breeding mice for 2 weeks, blood was collected and serum was separated by centrifugation. Total cholesterol and LDL cholesterol were measured using a cholesterol measurement kit of sigma. In the group treated with pine needles extract and bark extract, it was confirmed that the total cholesterol level in serum was decreased and that the amount of LDL cholesterol was decreased. This means that the substance activating IDH3 alpha reduces the total cholesterol and LDL cholesterol levels in the blood, thereby treating and preventing atherosclerosis and cardiovascular disease.

Examples of the formulation of the composition will be described below, but the present invention is not intended to be limited thereto, but is intended to be described in detail. In the following, the active ingredient is at least one selected from the group consisting of one or more extracts selected from the group consisting of liver extract, rake extract, silk extract, Ulleung chrysanthemum extract, pine tree extract, Paddungnamul extract.

Formulation Example 1 Soft Capsule

An active ingredient 80 mg, soybean oil 180 mg, palm oil 2 mg, vegetable hardened oil 8 mg, lead 4 mg and lecithin 6 mg were mixed and filled with 400 mg per capsule according to a conventional method to prepare a soft capsule.

Formulation Example 2 Tablet

80 mg of active ingredient, 200 mg of galactooligosaccharide, 60 mg of lactose and 140 mg of maltose were mixed and granulated using a fluidized bed dryer, and then, 6 mg of sugar ester was added to a tablet press. The final weight of the content was 600 mg.

Formulation Example 3 Granules

80 mg of active ingredient, 250 mg of anhydrous glucose and 550 mg of starch were mixed, molded into granules using a fluidized bed granulator, and then filled into fabric. The final weight of the content was 1 g.

[Formulation Example 4] Drinks

80mg of active ingredient, 10g of glucose, 0.6g of citric acid and 25g of liquid oligosaccharides are mixed, and 300ml of purified water is added to each bottle to make 200ml. After filling the bottle sterilized for 4 to 5 seconds at 130 ℃ to prepare a beverage.

Formulation Example 5 Caramel Formulations

Caramel mold was formed by mixing 80 mg of active ingredient, corn syrup 1.8 g, skim milk 0.5 g, soy lecithin 0.5 g, butter 0.6 g, vegetable hardened milk 0.4 g, sugar 1.4 g, margarine 0.58 g and salt 20 mg. The final weight of the content was 6 g.

Formulation Example 6 Diet Bar Formulation

Caramel molding was performed by mixing 80 mg of active ingredient, 20 g of starch, 9 g of wheat starch, 11 g of starch syrup, 11.6 g of malt, 6 g of margarine, 30 mg of salt, 30 mg of citric acid, 140 mg of sodium bicarbonate and 2 g of sugar ester. The final weight of the content was 60 g.

As described above, the composition containing the plant extract according to the present invention serves to activate the TCA circuit by inducing the activity of the IDH3α gene promoter, thereby promoting the oxidative degradation of lipids. Thus, the plant extract containing composition according to the present invention is effective in the treatment or prevention of obesity, and is also effective in the treatment or prevention of hyperlipidemia.

Claims (6)

delete Composition for the prevention or treatment of obesity, containing one or more extracts selected from the group consisting of gangan extract, rake extract, vulgaris extract, Ulleung chrysanthemum extract and Paddungmul extract as an active ingredient. A composition for preventing or treating hyperlipidemia, which contains one or more extracts selected from the group consisting of soybean extract, rake extract, wild extract, Ulleung chrysanthemum extract, and Paddungnamul extract as active ingredients. The composition of claim 2 or 3, wherein the composition activates the tricarboxylic acid cycle in the liver. 5. The composition of claim 4, wherein the composition increases the activity of an isonate dehydrogenase enzyme (IDH3α) gene promoter. The composition of claim 2 or 3, wherein the extract is an extract of ethanol or methanol.

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