KR20190071270A - Anti-obesity composition comprising pepper leaf extract produced by ultra high pressure homogenization as effective component - Google Patents

Abstract

The present invention relates to a composition for anti-obesity containing a pepper leaf extract obtained through an ultra-high pressure homogenization process as an active ingredient. More particularly, the extraction yield and the total polyphenol content of the pepper leaf extract manufactured by the manufacturing method comprising an ultra-high pressure homogenization step are increased than those without the ultra-high pressure homogenization process. Since the extract has an excellent effect of inhibiting fat accumulation, the extract can be usefully used as a health functional food for preventing or alleviating obesity or as a therapeutic agent for obesity.

Description

[0001] The present invention relates to an anti-obesity composition comprising pepper leaf extract obtained by an ultra-high pressure homogenization process as an active ingredient,

The present invention relates to an anti-obesity composition containing an extract of a green leaf obtained through an ultra-high pressure homogenization process as an active ingredient, and more particularly, to a composition for an anti-obesity containing an extract of a green leaf obtained through an ultra high pressure homogenization process which can be used as a health functional food composition or a pharmaceutical composition The present invention relates to a composition for preventing, ameliorating or treating obesity contained as an active ingredient.

Obesity is a metabolic disorder caused by an imbalance in the intake and consumption of calories, and refers to a state of abnormally increased fat tissue due to excessive calories. Recently, the population of obese people is rapidly increasing due to the decrease in eating habits and physical activity. Obesity is a cause of various diseases such as hypertension, elevated blood cholesterol, diabetes, kidney disease, stroke, arteriosclerosis, fatty liver, arthritis, cancer, sleep and apnea.

Obesity is caused by accumulation of triglyceride (TG) in the adipocyte by differentiation and adipogenesis of preadipocyte, and regulating this lipogenesis mechanism is called obesity inhibition It is known as an effective treatment method. However, existing obesity remedies are used only by a small number of obese patients because of their ineffective effect or various side effects. The appetite suppressant that acts on the central nervous system includes the noreadrenaline drug phentermine, the serotonin drug fluoxetine, the noreadrenaline drug and the serotonin drug sibutramine, Orlistat, the endogenous cannabinoid receptor antagonist rimonabant, and taranabant. However, the above-mentioned obesity treatment drugs have side effects such as nausea, dizziness, diarrhea, constipation and hypertension due to mental disorders such as depression, anxiety and insomnia. In particular, rimonabant and sibutramine are not commercially available due to side effects At present, only orlistat, belviq and qsymia are approved by the FDA. Therefore, it is required to develop a drug for obesity which has a high anti-obesity effect and a low side effect among natural products expected to minimize the possibility of a safety problem.

On the other hand, the pepper leaves are composed of 79.4% moisture, 4.1% protein, 1% lipid, 3.8% cellulose, 8.2% carbohydrate and 3.5% ash and contains abundant vitamin A, vitamin C, polyphenol and flavonoids. DPPH Radical scavenging ability, hydrogen peroxide scavenging ability and high antioxidant ability. In addition, it is known that it has an anticancer effect such as breast cancer, colorectal cancer and stomach cancer, cancer cell proliferation inhibitory effect, complement system activity effect, immunity enhancement, antimicrobial activity and tyrosinase inhibitory activity. These leafy leaves are used for some edible purposes, but most of the leaves and stems are discarded. Since pepper leaves contain useful substances with no spicy taste unlike pepper pulp, it is a valuable resource for industrial use because it can develop high value-added products with high functionality when developing functional materials using them.

On the other hand, studies for improving the efficacy and extraction yield of the materials reported so far have shown improvements in yield due to increase in surface area due to fine grinding in the physical aspect, application of ultrasonic waves and homogenization for cell disruption, increase in solubility according to the polarity of the solvent And biologically, fermentation and enzyme treatment processes for the solubilization of insoluble materials have been reported. However, the ability of the material to improve the extraction efficiency and yield has been limited.

The ultrahigh pressure homogenization technique is applied to the dispersing solution by passing through the micro-orifice module under high pressure (100 MPa or more) pressure, and by rapid increase of physical energy due to cavitation and high shear, It is a technique to convert a dispersion solution into a homogeneous phase by deagglomeration and disassociation of the dispersions in solution (depolymerization, low molecular weight). Particularly, it is known that the fiber is made into a homogeneous dispersed phase and is effective for low molecular weight of protein and non-aggregated carbohydrate polymer.

Korean Patent Laid-Open Publication No. 2004-0095949 discloses a composition for inhibiting obesity including plant leaves, Korean Patent Publication No. 2008-0114454 discloses a composition for inhibiting obesity using a Cheongyang pepper extract, An anti-obesity composition containing an extract of a green leaf obtained through an ultra-high pressure homogenization process as an active ingredient has not been disclosed.

The present invention provides an anti-obesity composition containing an extract of a green leaf extract obtained through an ultra-high pressure homogenization process as an active ingredient, and has an effect of inhibiting lipogenesis of a green leaf extract extracted through an ultra-high pressure homogenization process And that the extraction yield and the total polyphenol content were higher than those obtained without the ultra-high pressure homogenization process, thereby completing the present invention.

In order to solve the above problems, the present invention provides a health functional food composition for preventing or ameliorating obesity, which comprises an extract of green leafy leaves produced by a method comprising the following steps:

(1) drying a green leaf and then pulverizing the green leaf to produce a green leaf powder;

(2) adding an extraction solvent to the peppery leaf powder prepared in the step (1) and stirring the mixture;

(3) centrifuging and extracting the extract of step (2) to prepare a filtrate; And

(4) After the step (3), the filtrate is subjected to ultra-high pressure homogenization.

In addition, the present invention provides a pharmaceutical composition for preventing or treating obesity containing the above-described green leaf extract as an active ingredient.

The present invention relates to an anti-obesity composition containing an extract of a green leaf obtained through an ultra-high pressure homogenization process as an active ingredient. The extract of the green leaf extract obtained through the ultra-high pressure homogenization process of the present invention has a higher total polyphenol content The extraction yield is increased and the fat production is inhibited, thereby exhibiting an anti-obesity effect. Therefore, the composition of the present invention comprising the extract of green leafy which is obtained through the ultra-high pressure homogenization process as an active ingredient can be used as a material for prevention, improvement or treatment of obesity.

FIG. 1 shows the result of confirming the cytotoxicity of the green leaf extract prepared according to the method of the present invention. CON, a green tea leaf extract-free treated with a green tea leaf extract, Comparative Example 1, a pepper leaf extract, a Pectinex enzyme-treated green leaf extract, a Pectinex enzyme treatment and an ultra-high pressure homogenization treatment; And Experimental Example 1 is a green leaf extract through an ultra-high pressure homogenization process.
Fig. 2 shows the result of confirming the degree of inhibition of lipogenesis of the green leaf extract prepared according to the method of the present invention. MDI (a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin) was used as a differentiation inducing substance to induce the differentiation of 3T3-L1 as a lipogenic precursor cell. CON, MDI and green leaf extract-free control, Comparative Example 1, Ethanol extract of pepper leaf extract, Comparative Example 2, Pectinex enzyme-treated green leaf extract, Comparative Example 3, Pectinex enzyme treatment and Extra-high pressure homogenization; And Experimental Example 1 is a green leaf extract through an ultra-high pressure homogenization process.

The present invention

(1) drying a green leaf and then pulverizing the green leaf to produce a green leaf powder;

(2) adding an extraction solvent to the peppery leaf powder prepared in the step (1) and stirring the mixture;

(3) centrifuging and extracting the extract of step (2) to prepare a filtrate; And

(4) a step of subjecting the filtrate to ultra-high pressure homogenization after the step (3), and a health functional food composition for preventing or ameliorating obesity containing the extract of green tea leaf as an active ingredient.

The extraction solvent added in the step (2) is preferably a C ₁ to C ₄ lower alcohol, water or a mixture thereof, more preferably ethanol, but is not limited thereto.

The ultra-high pressure homogenization in the step (4) of the present invention is performed by repeating the process of passing through the orifice tube at a pressure of 100 to 120 MPa for 1 to 3 times, preferably passing through the orifice tube at a pressure of 110 MPa twice, I never do that.

As a preferred example of the present invention,

(1) drying a green leaf and then pulverizing the green leaf to produce a green leaf powder;

(2) adding 40 parts by weight of 70% (v / v) ethanol to 1 part by weight of the green leaf powder prepared in the step (1), followed by stirring and extracting;

(3) centrifuging and extracting the extract of step (2) to prepare a filtrate; And

(4) After the step (3), the filtrate is passed through an orifice tube twice under a pressure of 110 MPa to prevent or improve the obesity containing the extract of the green leafy leaf as an active ingredient Health functional food composition, but are not limited thereto.

In the health functional food composition according to one embodiment of the present invention, the health functional food composition can inhibit fat accumulation, but is not limited thereto.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method. The amount of the active ingredient can be suitably used depending on its use purpose (prevention or improvement). Generally, the health functional food composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight based on the raw material, when the food or beverage is produced. However, in the case of long-term consumption intended for health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount of more than the above range.

There is no particular limitation on the kind of the health functional food. Examples of the foods to which the health functional food composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, soups, Drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

In addition, the health functional food composition of the present invention can be produced as a food, particularly a functional food. The functional food of the present invention includes components that are ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when it is made of a drink, it may contain, in addition to the active ingredient, a natural carbohydrate or a flavoring agent as an additional ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the health functional food composition, it is also possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like. Although the ratio of the above-mentioned ingredients is not critical, it is generally selected in the range of 0.01 to 0.1 part by weight based on 100 parts by weight of the health functional food composition of the present invention.

In addition,

(1) drying a green leaf and then pulverizing the green leaf to produce a green leaf powder;

(2) adding an extraction solvent to the peppery leaf powder prepared in the step (1) and stirring the mixture;

(3) centrifuging and extracting the extract of step (2) to prepare a filtrate; And

(4) a step of subjecting the filtrate to ultra-high pressure homogenization after the step (3), and a pharmaceutical composition for preventing or treating obesity containing the extract of green tea leaf as an active ingredient.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier in addition to the active ingredient. Such a carrier is usually used at the time of formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, Calcium carbonate, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals Oils, and the like, but are not limited thereto. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences, 19th ed., 1995.

The appropriate dosage of the pharmaceutical composition according to the present invention may vary depending on such factors as the formulation method, the administration method, the patient's age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, .

The pharmaceutical composition of the present invention may be administered orally or parenterally. In the case of parenteral administration, the composition may be administered topically to the skin, intravenously, subcutaneously, intramuscularly, intraperitoneally, or transdermally.

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the inhibition and treatment of obesity.

The concentration of the active ingredient contained in the composition of the present invention can be determined in consideration of the purpose of treatment, the condition of the patient, the period of time required, and the like, and is not limited to a specific range of concentration.

The pharmaceutical composition of the present invention may be manufactured in a unit dosage form by formulating it using a pharmaceutically acceptable carrier or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs Into a capacity container. The formulations may be formulated into any one of the formulations selected from injectables, creams, patches, sprays, ointments, alerts, lotions, liniments, pastes and cataplasms, and may additionally contain dispersing or stabilizing agents have.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples and Examples. It is to be understood by those skilled in the art that these preparations and examples are merely intended to explain the present invention more specifically and that the scope of the present invention is not limited thereto.

Manufacturing example  1. Preparation of green leaf extract

(1) Preparation of Extra-high pressure homogenized green leaf extract

100 ml of 70% (v / v) ethanol prepared by using distilled water adjusted to pH 4.5 was added to 2.5 g of dried and pulverized green leaf, and the mixture was stirred at room temperature for 180 minutes at 180 rpm for extraction. The extracted extract was centrifuged at 12,000 rpm for 15 minutes, filtered to prepare a filtrate, and then passed through an orifice tube twice under a pressure of 110 MPa to prepare an ultra-high pressure homogenized green leaf extract. The extract was used as Experimental Example 1.

(2) Preparation of ethanol extract of hot pepper leaves

100 ml of 70% (v / v) ethanol prepared by using distilled water adjusted to pH 4.5 was added to 2.5 g of dried and pulverized green leaves, extracted with stirring at 180 rpm at room temperature for 2 hours, and then extracted at 12,000 rpm for 15 minutes Centrifuged and filtered to prepare ethanol extract of hot pepper, which was used as Comparative Example 1 of the present invention.

(3) Preparation of Pectinex enzyme-treated green leaf extract

30 ml of distilled water adjusted to pH 4.5 and 500 units / ml of pectinex were added to 2.5 g of dried and pulverized green leaves, and the mixture was stirred at 37 ° C for 2 hours at 180 rpm. Thereafter, 70 ml of 100% (v / v) ethanol was added and the mixture was extracted at 180 rpm for 2 hours at room temperature. The mixture was centrifuged at 12,000 rpm for 15 minutes and filtered to prepare an enzyme-treated green leaf extract.

(4) Preparation of Pectinex enzyme treatment and high pressure homogenization parallel treatment of green leaf extract

30 ml of distilled water adjusted to pH 4.5 and 500 units / ml of pectinex were added to 2.5 g of dried and pulverized green leaf, and the mixture was stirred at 37 ° C for 2 hours at 180 rpm. Thereafter, 70 ml of 100% (v / v) ethanol was added thereto, stirred and extracted at 180 rpm for 2 hours at room temperature, centrifuged at 12,000 rpm for 15 minutes and filtered to prepare a filtrate. After passing through the orifice tube twice, the Pectinex enzyme treatment and the high pressure homogenization treatment were carried out. The extract was used as Comparative Example 3.

Example  1. Comparison of yield of green leaf extract according to extraction method

The extraction yields of the green leaf extracts extracted for each condition in Production Example 1 were compared. As a result, as shown in Table 1, extraction yield was increased when enzyme treatment, ultra-high pressure homogenization, or enzyme treatment and ultrahigh pressure homogenization were performed in comparison with Comparative Example 1, which is an ethanol extract of a leafy green leaf. The extraction yield was calculated by the following formula 1, and the increase rate was the degree of increase with reference to the comparative example 1. [

[Formula 1]

Extraction yield (%) = (weight of extracted powder (g) / weight of initial sample (g)) x 100

Treated group Extraction yield (%) Growth rate (%) Comparative Example 1
(Ethanol extraction) 31.60 - Comparative Example 2
(Pectinex treatment) 50.58 60.06 Comparative Example 3
(Ultra-high pressure homogenization + pectinex processing) 51.15 61.87 Experimental Example 1
(Ultra-high pressure homogenization treatment) 37.60 18.99

Example  2. Changes in polyphenol contents of green leaf extracts by extraction method

In Preparation Example 1, the total polyphenol content of the green leaf extract extracted by each condition was confirmed. As a result, as shown in Table 2, when the enzyme treatment, the ultrahigh pressure homogenization or the enzyme treatment and the ultrahigh pressure homogenization were performed in parallel, it was confirmed that the total polyphenol content was increased compared to the ethanol extract of the pepper leaf of Comparative Example 1.

Treated group Polyphenol content (mg / extract 1 g) Comparative Example 1
(Ethanol extraction) 36.50 Comparative Example 2
(Pectinex treatment) 40.39 Comparative Example 3
(Ultra-high pressure homogenization + pectinex processing) 40.10 Experimental Example 1
(Ultra-high pressure homogenization treatment) 37.61

Example  3. Cell viability ( cell viability ) Measure

In order to examine the effect of the green leaf extract on the cell survival of each condition in Production Example 1, MTT (3- (4,5-dimeth-ylthiazol-2-yl) -2,5 -diphenyltetrazolium bromide) analysis. 3T3-L1, a lipogenic precursor cell, was cultured in a 96-well plate containing 100 μl of DMEM medium for 1 day and then treated with 320 ppm of a green leaf extract and cultured for 24 hours. Then, 10 μl of 5 mg / ml MTT solution was added to each well, followed by culturing for 3 hours to induce a reduction reaction. 100 μl of DMSO (dimethyl sulfoxide) solution was added to form purple formazan ) Crystals were completely dissolved. After that, the absorbance was measured at 570 nm using a spectrophotometer, and the relative cell viability of the group treated with the green leaf extract was calculated based on the survival rate of 100% of the green leaf extract-free group.

As a result, as shown in Fig. 1, it was found that the extract of the green tea leaves of each of the conditions of Preparation Example 1 did not significantly affect the cell viability, indicating that there was almost no cytotoxicity.

Example  4. Inhibitory Effect of Extracts from Extracts

The lipogenesis of 3T3-L1, a lipid precursor cell, was measured by staining with a lipid (Oil Red-O) staining kit (Sigma, USA). The extract was treated with 320 ppm of the green leaf extract of Preparation Example 1 and the differentiation inducer (MDI) to differentiate and then treated with 10% formalin-buffered phosphate buffer (PBS), fixed at room temperature, After washing with isopropanol, it was dried. After that, Oil Red-O was added, stained for 45 minutes, washed 4 times with distilled water, and the distilled water was completely removed. After that, 100% isopropanol was added to the cells, and Oil Red-O stained with fat was extracted for 10 minutes at 180 rpm, and the value was measured using absorbance at 520 nm.

As a result, as shown in Fig. 2, the fat-generation inhibitory effect was not observed in the ordinary ethanol extract (Comparative Example 1) and the enzyme alone treatment group (Comparative Example 2) (Comparative Example 3) in which the treatment with pectinex treatment and ultra-high pressure homogenization was performed in parallel, showed no inhibitory effect on lipogenesis.

Claims (6)

A health functional food composition for preventing or ameliorating obesity containing an extract of green tea leaf produced by a method comprising the following steps as an active ingredient:
(1) drying a green leaf and then pulverizing the green leaf to produce a green leaf powder;
(2) adding an extraction solvent to the peppery leaf powder prepared in the step (1) and stirring the mixture;
(3) centrifuging and extracting the extract of step (2) to prepare a filtrate; And
(4) After the step (3), the filtrate is subjected to ultra-high pressure homogenization. The health functional food composition according to claim 1, wherein the extraction solvent in step (2) is a C ₁ -C ₄ lower alcohol, water or a mixture thereof. The health functional food composition for preventing or improving obesity according to claim 1, wherein the ultra-high pressure homogenization of step (4) is performed at a pressure of 100 to 120 MPa. The health functional food composition for preventing or improving obesity according to claim 1, wherein the ultra-high pressure homogenization of step (4) is repeated one to three times. The health functional food composition according to claim 1, wherein the green leaf extract inhibits fat accumulation. A pharmaceutical composition for preventing or treating obesity comprising an effective component of a green leaf extract prepared by a method comprising the steps of:
(1) drying a green leaf and then pulverizing the green leaf to produce a green leaf powder;
(2) adding an extraction solvent to the peppery leaf powder prepared in the step (1) and stirring the mixture;
(3) centrifuging and extracting the extract of step (2) to prepare a filtrate; And
(4) After the step (3), the filtrate is subjected to ultra-high pressure homogenization.

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