KR101007088B1 - Fatty acid oxidation-promoting composition containing herbal and native plant extracts - Google Patents

Abstract

Fatty acid oxidation promoting composition according to the present invention is a human extract by containing at least one selected from the group consisting of extracts, white porcelain extract, white root extract, Jeonho extract, Cheonphu extract, soybean extract, Paekkuk extract and bamboo shoot extract as an active ingredient, Increasing the activity of the Carnitine Palmitoyl Transferase 1A (CPT1A) promoter, a gene that plays an important role in fatty acid oxidation control in hepatocytes, increases the expression of the CPT1A gene in the liver, thereby promoting the oxidation of fatty acids in the liver. It can be used for the prevention or treatment of obesity, the prevention or treatment of hyperlipidemia.

Chinese cabbage, baekja, baekbukeun, Jeonho, Cheon-oh, Padu, Paek-geun, Bamboo, Fatty Acid, Oxidation, CPT1A, Obesity, Hyperlipidemia

Description

Composition for promoting oxidation of fatty acid comprising extract of crude drugs and native plants}

1 is a human extract of the human liver cancer cell line Huh7 cells containing a CPT1A promoter and luciferase fusion structure, respectively, when the extracts of Doin, Baekja extract, Paekjakeun extract, Jeonho extract, Cheon oh extract, Padu extract, Pajanggeun extract, It is a graph showing the activity of luciferase.

Figure 2 is a graph showing the oxidation amount of fatty acids when treated with Hein extract, white extract, white root extract, Jeonho extract, Cheonphu extract, Padu extract, Paekkuk extract and bamboo shoot extract to HepG2 cells, human liver cancer cell line.

The present invention relates to a composition containing a herbal and autologous plant extract that functions to promote the oxidation of fatty acids, and more particularly, to CPT1A (Carnitine) which is a gene that plays an important role in the regulation of fatty acid oxidation in human hepatocytes. Palmitoyl Transferase 1A) increases the activity of the promoter to increase the expression of the CPT1A gene in the liver and thereby promotes the oxidation of fatty acids in the liver. The present invention relates to a composition containing the self-derived deadwood extract and its use.

Carnitine palmitoyl transfrase 1A (hereinafter referred to as “CPT1A”) enzyme is a protein present in the mitochondria in cells and serves to deliver long chain fatty acids into the mitochondria. The enzyme is known to be involved in the rate determining step in the oxidation of long chain fatty acids in the mitochondria in cells.

In cell-level and animal-level experiments, it has been found that increasing the expression of CPT1A promotes oxidation of fatty acids and increases energy consumption. The treatment of hyperlipidemia induced by high-fat diet was improved when ZTA rats, which induced obesity through the administration of high-fat diet, were treated with TTA (Tetradecylthioacetic acid), a drug that promotes fatty acid beta oxidation by increasing CPT1 expression. Increasing susceptibility has been known (CT Cramer et al. Progress in Lipid Research , 40 , pp231-268, 2001), and ragaglitazar, which has been shown to increase the activity of CPT1 in the liver, was treated in high-fat diet hamsters. It has been shown to reduce the amount of fat (R. Chakrabarti et al. British Journal of Pharmacology , 140, pp 527-537, 2003).

In light of these results, it is expected that substances that increase the expression of CPT1 may contribute to the treatment and prevention of obesity and hyperlipidemia by promoting the oxidation of fatty acids in cells, increasing energy consumption and reducing the amount of fat in the body.

CPT1 is known to exist in two forms, CPT1A, which exists in hepatocytes, and CPT1B, which exists in muscles. Gene expression of CPT1 in animals and cells is regulated by cellular and hormonal conditions. Increasing glucagon and decreasing insulin increase the expression of the CPT1 gene, and it is known that eicopentaenoic acid (EPA) in fish oil increases the expression of CPT1 and promotes oxidation of fatty acids.

Therefore, the present inventors searched for hundreds of herbal and native plant extracts to find plant extracts that increase the expression of CPT1A gene in hepatocytes and promote the oxidation of fatty acids. The present invention was completed by confirming that the extracts of the roots and the extracts of native bamboo shoots, which increase the activity of the promoter for the expression of the CPT1A gene in hepatocytes, promote the oxidation of fatty acids.

Accordingly, it is an object of the present invention to provide a composition for promoting fatty acid oxidation, a composition for preventing or treating obesity, a composition for preventing or treating hyperlipidemia, which is effective in preventing or treating obesity and hyperlipidemia by promoting oxidation of fatty acids in the liver.

In order to achieve the above object, the composition for promoting fatty acid oxidation according to the present invention is one selected from the group consisting of Doin extract, Baekja extract, Paekkukeun extract, Jeonho extract, Cheon oh extract, Padu extract, Pajanggeun extract, and bamboo shoot extract It is characterized by containing the above extract as an active ingredient.

In addition, the composition for preventing or treating obesity according to the present invention contains one or more extracts selected from the group consisting of cabbage extract, white baekja extract, white root extract, jeonho extract, green onion extract, janggeun extract and bamboo shoot extract as an active ingredient. It features.

In addition, the composition for preventing or treating hyperlipidemia according to the present invention contains one or more extracts selected from the group consisting of Doin extract, Baekja extract, Paikeun extract, Jeonho extract, Padu extract, Paekjanggeun extract and bamboo shoot extract as an active ingredient. It features.

The composition for preventing or treating obesity or hyperlipidemia preferably further comprises a cheonho extract.

In any one of the above compositions according to the invention, the composition preferably increases the expression of the CPT1A gene by increasing the activity of the CPT1A gene promoter in the liver.

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

Most preferably the ethanol is 95% ethanol.

In any one of the above compositions according to the invention, the composition may be a pharmaceutical composition or health functional food composition.

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

Persicae Semen is a deciduous arborescent tree belonging to the Rosaceae family and is the core of mature fruit of peach or mountain copy. It grows or grows in wild fields all over Korea. It is 6m high and flowers are light red in April ~ May. It blooms first, the medicinal form is a flat, uneven left and right oval shape, one end is sharp and the other is round, and there is a confluence here. Written and sweet in nature, without poison. It is used mainly for menstrual disorders and dysmenorrhea as it actively moves the blood and removes blood.

The Baekja ( Sinapis Semen ) is a mature seed of mustard and mustard, a herbaceous or young herbaceous herb that belongs to the cruciferous family. It is cultivated all over Korea. The medicinal form is 1.5 ~ 2.5mm in diameter and the surface is pale yellow. Is thin and brittle. Stop coughing or asthma and stop the swelling, stuffiness and soreness of the chest, which is effective for chronic bronchitis or bronchial expansion.

Stemonae Radix is a vine root of perennial herb, a perennial herb, belonging to the encyclopedia . It is not found in Korea and grows or grows in all regions except the western part of China. 60 ~ 90cm high, roots thick and fusiform, dozens gather. Medicinal herbs are usually waterproof, 4 ~ 18cm long, 7 ~ 10mm in diameter, straight or slightly curved, and thin at both ends. Has a very good effect on cough and asthma.

Jeonho ( Peucedani Radix ) is a dried root of the main body of the Araaceae family, and the roots of the scarlet bodice. It grows in the alpine region of Korea, has a height of 1m, the root is thick, and the trunk is hollow and splits from the top. The herb is fusiform and stems and leaves are left in the head. Jeonho is a medicine that is effective in removing coughs and stopping coughs.

Cheon-oh ( Aconiti Tuber ) is a dried tuber of Odu belonging to the buttercup family, dried in the sun from late June to early July, dried in the sun, 1.5 ~ 3cm long, and the surface is grayish brown and wrinkled. Cheon-O is used to eliminate the scams, keep the meridians warm, stop the pain, acute rheumatoid polyarthritis, swelling and soreness of the limbs, soreness, paraplegia, and swelling of the stomach and the stomach.

Padu ( Tiglii Semen ) is a dried fruit of Padu, belonging to the Great Fruit family. It grows or grows in southern China, and its height is 5m and leaves are displaced. The medicinal herb is flat and elliptical, and the outer surface of the epidermis is dark reddish brown to grayish brown with black spots. Padu is a spicy and poisonous herb that has the property of clearing and blocking the jangjangyuk.

Patriniae Radix , also known as matari , is a perennial herb of the dicotyledonous plywood grouped in the horned tree, Matariaceae , which is distributed from north to south of the Japanese archipelago in Taiwan, China and eastern Siberia, and is about 60-150 cm high and has a thick root stem. And stretches to the side. Flowers bloom yellow and bloom from summer to autumn. Use mild shoots as herbs and use outpost as anti-inflammatory, fish-blood or pus-free medicine.

Styrax japonica is a deciduous arborescent tree of the dicotyledon plant Plywood group, Prunus japonica , which is distributed in Korea (Southern Central), Japan, the Philippines, and China, and its height is around 10. There is a virgin on the branch, but it disappears and the epidermis is peeled off and becomes dark brown. Leaves are alternate, egg-shaped or long oval, flat or slightly serrated. The flowers are unisexual and have a bell shape. When the bamboo extract contained in the composition according to the present invention can use all of the leaves, the bark of the stem, the heart of the stem, the fruit is not particularly limited, it is most preferred to use the fruit.

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 a shade at room temperature, and then 40 to 60 ° C. using ethanol, preferably 95% ethanol or methanol, butanol, ethyl acetate, chloroform, or hexane, 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 was added to each active fraction content, homogeneously suspended, and then lyophilized to obtain powdered active fractions. Can be. 95% ethanol is preferably used as an extraction solvent for extracts of Chinese herbs, Baekja, Paekkukeun, Jeonho, Cheonho, Padu and Paenggeun, and methanol extract is preferred for extracts of wild bamboo, which are native plants. .

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.

In order to determine the effect of promoting the CPT1A promoter activity of each of the extracts, the present inventors treated Huh7 cells, which are human liver cancer cell lines containing the CPT1A promoter-luciferase fusion gene, with the respective extracts to measure the activity of luciferase. As a result of measuring activity, the effect of increasing luciferase activity was confirmed.

In addition, in order to determine the effect of increasing the fatty acid beta oxidation by the extract, as a result of measuring the beta oxidation by quantifying the ³ H ₂ O contained in the medium by treating the human liver cancer cell line HepG2 cells with each of the extracts, fatty acid beta oxidation The increase effect was confirmed.

The composition according to the present invention is any one or more selected from the group consisting of Doin extract, white back extract, white root extract, Jeonho extract, Cheonphu extract, Padu extract, Paekkuk extract and Staphylococcus extract, which are active ingredients, based on the total weight of the composition It is preferable to contain in quantity of 0.0001 to 99.9 weight%. 0.1-60.0% by weight is more preferable in view of 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 skill in the art and generally dosages range 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, sol, 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 the extract and a food supplement acceptable food supplement. Examples of the food to which the extract can be added 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 may be added at 0.001 to 99.9% by weight of the total food weight, the health beverage composition is based on 100 ml in a ratio of 0.001 to 0.1 g, more preferably 0.05 to 0.1 g Can be added.

The health functional beverage composition of the present invention has no particular limitation on the other ingredients other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary 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 in 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 the following Examples and Experimental Examples, but the present invention is not limited to these Examples and Experimental Examples.

Experimental Example 1. Analysis of CPT1A promoter activity of herbal and native plant extracts

<1-1 steps. Cell Lines and Cell Cultures>

In step 1-1, Huh7 cells (see prior patent publication No. 2004-0060749), which are human hepatoma cell lines containing the CPT1A promoter-luciferase fusion gene, are treated with 10% bovine serum (fetal bovine). 10 ml / flask of DMEM medium (Dulbecco's modifided Eagle's Medium, Gibco 1210-0038) containing the cells was inoculated with 1.0 × 10 ⁶ cells and cultured at 37 ° C. in a 5% CO ₂ incubator.

<1-2 steps. Increased Activity of CPT1A Promoter by Herbal and Natural Plant Extracts>

Persicae Semen Extract (Par. No .: CA01-035), Sinapis Semen Extract (Par. No .: CA01-064), Stemonae Radix Extract (Par. No., obtained from Korea Plant Extract Bank, Daejeon, Korea) : CA01-070), Peucedani Radix Extract (Par. No .: CA03-049), Aconiti Tuber Extract (Par. No .: CA04-051), Pungdu ( Tiglii Semen ) Extract (Par .: CA03-079) , Patriniae Radix extract (preparation No .: CA04-073), when using the jukki extract (preparation No .: 001-064) was performed as follows. At this time, each extract of the dry powder state was dissolved in DMSO (dimethyl sulpholxide) solvent and treated to cells.

Huh7 cells cultured in step 1-1 were each treated with 3 ml / flask of trypsin to make a single cell suspension and incubated in a 96 well plate at 1.0 × 10 ⁴ cells / well and incubated for 24 hours. Thereafter, DMEM medium without bovine serum was treated with 100 microL / well and incubated for 24 hours (control). Then, in the 100 microL / well medium containing no bovine serum, the prepared Doin extract, Baekja extract, Paek-geun extract, Jeonho extract, Cheon oo extract, Padu extract, Paeng-geun-geun, and bamboo shoots were added at a concentration of 10 ppm each. The cell culture was treated for 24 hours. After treatment, the activity of the promoter was measured by measuring the activity of luciferase expressed using a Luciferase assay kit (promega, USA). Luciferase activity was measured by the same method as above for the control group to compare the effect of each extract according to the present invention. The results are shown in FIG.

As shown in Figure 1, compared with the control group, the extract of the present invention, white dog extract, white root extract, Jeonho extract, Cheonphu extract, green onion extract, Paekkukeun, sichuan tree extract 2.1 times, 2.7 times, 2.4 times, 2.5 times, 2.6 times, 2.5 times, 2.3 times, 3.0 times, it was confirmed that there is an effect of increasing luciferase activity.

Experimental Example  2. Increased fatty acid beta oxidation by herbal and native plant extracts

<2-1 step. Cell Lines and Cell Cultures>

In step 2-1, HepG2 cells, a human hepatoma cell line, were 1.0 × 10 ⁶ cells in 10 ml / flask of DMEM medium (Gibco 1210-0038) containing 10% fetal bovine serum. Inoculation was incubated and all cultured in 37 ° C., 5% CO ₂ incubator.

<2-2 steps. Increased fatty acid beta oxidation by each extract>

HepG2 cells cultured in step 2-1 were each treated with 3 ml / flask of trypsin to make a single cell suspension, and the cells were incubated for 24 hours by dispensing 2 × 10 ⁵ cells / well in 6 well plates. Thereafter, DMEM medium without bovine serum was treated with 1 ml / well and incubated for 24 hours (control). Then also in 1ml / well of medium containing no serum, doin extract, white baekja extract, baekeungup extract, Jeonho extract, Cheonphu extract, Padu extract, Paekjanggeun, and bamboo shoots prepared in step 1-2 of Experimental Example 1 Each was added at a concentration of 10 ppm and treated for 24 hours in the cell culture. Genistein was treated with 10 microM as a positive control. After 24 hours after each extract and genistein treatment, the medium was removed and washed with PBS, and 1 mM L-carnitine and [9,10]-in 1ml / well of DMEM medium containing no bovine serum. ³ h palmitic acid 3 mCi / well ([9,10] ^-3 H palmitic acid 3 mCi / well) was added to react for 3 hours. After the reaction, the medium was collected, and fatty acid beta oxidation could be measured by quantifying ³ H ₂ O contained in the medium using a scintillation counter (Scintillation counter, 1450 MicroBeta, Wallac). The results are shown in FIG.

As shown in Figure 2, in the case of each treatment group of Doin extract, Baekja extract, Paekkugeun extract, Jeonho extract, Cheonphu extract, Padu extract, Paekjanggeun, and bamboo shoot extract according to the present invention significantly fatty acid beta oxidation compared to the control group It was confirmed that the increase, and showed a better effect than the positive control, Genistein.

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 Doin extract, Baekja extract, Paekbugeun extract, Jeonho extract, Cheonphu extract, Padu extract, Pajanggeun extract, and bamboo shoot 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, followed by tableting with a tablet press by adding 6 mg of sugar ester. The final weight of the content is 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 is 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 Formulation

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 is 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 is 60 g.

As described above, the composition containing one or more selected from the group consisting of the above-mentioned doin extract, white baekja extract, white root extract, Jeonho extract, Cheonphu extract, green onion extract, Paekkuk extract, and oleander extract according to the present invention is human hepatocyte By increasing the activity of the CPT1A promoter, a gene that plays an important role in fatty acid oxidation control in the state, it increases the expression of the CPT1A gene in the liver, thereby promoting the oxidation of fatty acids in the liver. Thus, the herbal and native 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 (7)

Fatty acid oxidation-promoting composition containing at least one extract selected from the group consisting of cabbage extract, white baekja extract, baekbugeun extract, Jeonho extract, Cheonphu extract, green onion extract, and oleander extract. Composition for the prevention or treatment of obesity, containing one or more extracts selected from the group consisting of Paekjakja extract, Paekkugeun extract, Padu extract, Paekkugeun extract, and bamboo shoot extract. A composition for the prevention or treatment of hyperlipidemia, which comprises at least one extract selected from the group consisting of white back extract, white root extract, Jeonho extract, green bean extract, and oleander extract. 4. A composition according to claim 2 or 3, wherein the composition further comprises a cheol extract. The composition of claim 1, wherein said composition increases CPT1A gene expression by increasing the activity of the CPT1A gene promoter in the liver. The composition according to any one of claims 1 to 3, wherein the extract is an extract of ethanol or methanol. The composition of any one of claims 1 to 3, wherein the composition is a pharmaceutical composition or a nutraceutical composition.

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