KR102034183B1 - Anti-obesity composition comprising pepper leaf extract produced by ultra high pressure homogenization as effective component - Google Patents

Abstract

The present invention relates to an anti-obesity composition containing the pepper leaf extract obtained through the ultrahigh pressure homogeneous process as an active ingredient, and more particularly, the extraction yield and the total polyphenol content of the pepper leaf extract prepared by the manufacturing method including the ultrahigh pressure homogeneous step. This ultra-high pressure homogeneous process is increased, and the extract is excellent in the effect of inhibiting fat accumulation, and thus can be usefully used as a dietary supplement for preventing or improving obesity or as a therapeutic agent for obesity.

Description

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

The present invention relates to an anti-obesity composition containing the pepper leaf extract obtained through the ultra-high pressure homogeneous process as an active ingredient, and more particularly to the pepper leaf extract obtained through the ultra-high pressure homogeneous process that can be used as a health functional food composition or a pharmaceutical composition. It relates to a composition for preventing, improving or treating obesity, which is contained as an active ingredient.

Obesity is a metabolic disease caused by an imbalance between calorie intake and consumption, and an abnormal increase in fat tissue due to excess calories. Recently, the obese population is rapidly increasing due to westernized eating habits and decreased 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, apnea rather than its own problems, it is important to prevent and treat obesity.

Obesity is caused by the accumulation of triglycerides (TG) in adipocytes by the process of differentiation and adipogenesis of preadipocytes. It is known to be an effective treatment. However, existing obesity treatments are only used by a small number of obese patients due to their ineffectiveness or various side effects. Appetite suppressants that act on the central nervous system include phentermine, a noradrenaline drug, fluoxetine, a serotonin drug, and sibutramine, a mixed drug with noradrenaline and serotonin. Phosphorus orlistat, an endogenous cannabinoid receptor antagonist rimonabant, taranabant and the like. However, the anti-obesity agent causes mental disorders such as depression, anxiety and insomnia, and side effects such as nausea, dizziness, diarrhea, constipation, and high blood pressure, in particular, rimonabant and sibutramine (sibutramine) stop marketing due to side effects. Currently only orlistat, belviq and qsymia are approved and commercially available from the FDA. Therefore, the development of anti-obesity drugs that have high anti-obesity effects and low side effects among natural products that are expected to minimize the possibility of occurrence of safety problems are required.

On the other hand, red pepper leaves are composed of 79.4% moisture, 4.1% protein, 1% lipid, 3.8% fibrin, 8.2% sugar, and 3.5% ash, and contain abundant vitamin A, vitamin C, polyphenols, flavonoids, and DPPH. It has been reported to have radical scavenging activity, hydrogen peroxide scavenging activity and high antioxidant activity. In addition, it is known to have anticancer effects such as cancer cell proliferation inhibitory effect, complement system activation effect such as breast cancer, colorectal cancer and gastric cancer, immune enhancement, antibacterial activity and tyrosinase inhibitory activity. The young shoots of these pepper leaves are used for some food, but most of the pepper leaves and stems are discarded. Red pepper leaves, unlike red pepper flesh, do not have a spicy taste and contain useful substances, so they can develop high-value, high value-added products when developing functional materials using them.

On the other hand, studies to improve the efficacy and extraction yield of the material reported so far, the physical improvement in the yield by the surface area increase according to the fine grinding, the use of ultrasonic and homogenizer for cell disruption, solubility according to the polarity of the solvent In the biological aspect, fermentation and enzyme treatment processes for solubilizing insoluble materials have been reported, but the efficacy of the material and the ability to improve extraction yields have been relatively limited.

Ultra-high pressure homogeneous technology disperses due to rapid increase in physical energy due to cavitation and high shear caused by rapid change of pressure by passing micro orifice module under pressure of dispersion solution at high pressure (over 100MPa). It is a technique that converts a dispersion into a homogeneous phase by deagglomeration or depolymerization of the dispersion in the solution. In particular, it is known that the fibers are made into a uniform dispersed phase and effective for low molecular weight of protein and non-starchy carbohydrate polymer.

On the other hand, Korean Patent Publication No. 2004-0095949 discloses a composition for inhibiting obesity including plant leaves, and Korean Patent Publication No. 2008-0114454 discloses an obesity inhibitory composition using the Chungyang pepper extract, There is no disclosure about an anti-obesity composition containing a pepper leaf extract obtained through an ultrahigh pressure homogeneous process as an active ingredient.

The present invention was derived by the above requirements, provides an anti-obesity composition containing the pepper leaf extract extracted through the ultra-high pressure homogeneous process as an active ingredient, the fat production inhibitory effect of the pepper leaf extract extracted through the ultra-high pressure homogeneous process is the most Excellent, complete the present invention by confirming that the extraction yield and total polyphenol content is increased than without the ultra-high pressure homogeneous process.

In order to solve the above problems, the present invention provides a health functional food composition for the prevention or improvement of obesity containing red pepper leaf extract prepared by a method comprising the following steps as an active ingredient:

(1) drying the red pepper leaves and then grinding to prepare red pepper leaf powder;

(2) adding the extraction solvent to the pepper leaf powder prepared in step (1), followed by stirring extraction;

(3) preparing a filtrate by centrifuging and extracting the extract of step (2); And

(4) after step (3), ultra-high pressure homogenizing the filtrate.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of obesity containing the pepper leaf extract as an active ingredient.

The present invention relates to an anti-obesity composition containing the pepper leaf extract obtained through the ultra-high pressure homogeneous process as an active ingredient, the pepper leaf extract extracted through the ultra-high pressure homogeneous process of the present invention has a total polyphenol content and The extraction yield increases and inhibits adipogenesis, resulting in an anti-obesity effect. Therefore, the composition of the present invention comprising a pepper leaf extract obtained through the ultra-high pressure homogeneous process as an active ingredient can be used as a material for the prevention, improvement or treatment of obesity.

1 is a result confirming the cytotoxicity of the pepper leaf extract prepared according to the method of extracting pepper leaves of the present invention. CON is red pepper leaf extract untreated negative control group, Comparative Example 1 pepper leaf ethanol extract, Comparative Example 2 pectinex enzyme treatment pepper leaf extract, Comparative Example 3 pepper leaf extract through pectinex enzyme treatment and ultra-high pressure homogenization process; And Experimental Example 1 is pepper leaf extract through the ultra-high pressure homogenization process.
Figure 2 is a result confirming the degree of inhibition of fat production of pepper leaf extract prepared according to the method of extracting pepper leaves of the present invention. MDI (a mixture of IBMX (3-isobutyl-1-methylxanthine), dexamethasone, and insulin) was used to induce differentiation of 3T3-L1, which is a progenitor cell. CON is MDI and pepper leaf extract untreated negative control group, Comparative Example 1 pepper leaf ethanol extract, Comparative Example 2 pectinex enzyme-treated pepper leaf extract, Comparative Example 3 pepper leaf extract through pectinex enzyme treatment and ultra-high pressure homogenization process; And Experimental Example 1 is pepper leaf extract through the ultra-high pressure homogenization process.

The present invention

(1) drying the red pepper leaves and then grinding to prepare red pepper leaf powder;

(2) adding the extraction solvent to the pepper leaf powder prepared in step (1), followed by stirring extraction;

(3) preparing a filtrate by centrifuging and extracting the extract of step (2); And

(4) after the step (3), and the high-pressure homogenization of the filtrate; relates to a health functional food composition for the prevention or improvement of obesity containing the pepper leaf extract prepared by the method comprising an active ingredient.

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

The ultra-high pressure homogenization of the step (4) of the present invention is to repeat the process of passing through the orifice tube 1 to 3 times under a pressure of 100 ~ 120MPa, preferably to pass through the orifice tube twice under a pressure of 110MPa but limited thereto I never do that.

One preferred embodiment of the present invention

(1) drying the red pepper leaves and then grinding to prepare red pepper leaf powder;

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

(3) preparing a filtrate by centrifuging and extracting the extract of step (2); And

(4) after the step (3), the filtrate is passed through an orifice tube twice under a pressure of 110MPa; for preventing or improving obesity containing the pepper leaf extract prepared by the method as an active ingredient It is a nutraceutical composition, but is not limited thereto.

In the health functional food composition according to an embodiment of the present invention, the health functional food composition may 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 used with other foods or food ingredients, and may be appropriately used according to a conventional method. The amount of the active ingredient may be appropriately used depending on the purpose of use (prevention or improvement). In general, in the preparation of food or beverages, the nutraceutical composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less with respect to the raw material. However, in the case of long-term intake for health purposes, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

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

In addition, the nutraceutical composition of the present invention may be prepared as a food, in particular a functional food. Functional foods of the present invention include ingredients that are commonly added in food production, and include, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when prepared with a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates can be monosaccharides (e.g. glucose, fructose, etc.), disaccharides (e.g. maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g. dextrins, cyclodextrins, etc.) or sugar alcohols (e.g. , Xylitol, sorbitol, erythritol and the like). The flavourant may be a natural flavourant (eg, taumartin, stevia extract, etc.) and a synthetic flavourant (eg, saccharin, aspartame, etc.).

Various nutritional supplements, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonic acid The carbonation agent etc. which are used for a drink can be contained further. Although the ratio of the above-mentioned ingredients is not critical, it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food composition of the present invention.

In addition, the present invention

(1) drying the red pepper leaves and then grinding to prepare red pepper leaf powder;

(2) adding the extraction solvent to the pepper leaf powder prepared in step (1), followed by stirring extraction;

(3) preparing a filtrate by centrifuging and extracting the extract of step (2); And

(4) after the step (3), and the high-pressure homogenization of the filtrate; relates to a pharmaceutical composition for the prevention or treatment of obesity containing the pepper leaf extract prepared by a method comprising as an active ingredient.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient, and such carriers are commonly used in the preparation of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, Calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals Oils and the like, but are not limited thereto. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences, 19th ed., 1995.

Suitable dosages of the pharmaceutical compositions according to the present invention may be variously prescribed by such factors as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and response to the patient. Can be.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered topically to the skin, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration and the like.

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 included 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, etc., and is not limited to a specific range of concentration.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier or excipient according to methods which can be easily carried out by those skilled in the art. It can be prepared by incorporation into a dose container. In this case, the formulation may be prepared in any one formulation selected from injection, cream, patch, spray, ointment, warning, lotion, linen, pasta and cataplasma, and may further include a dispersing or stabilizing agent. have.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples and Examples. These preparations and examples are only intended to explain the present invention more specifically, it will be apparent to those skilled in the art that the scope of the present invention is not limited thereto.

Production Example  1. Preparation of Red Pepper Leaf Extract

(1) Preparation of Ultra High Pressure Homogenized Red Pepper Leaf Extract

100 ml of 70% (v / v) ethanol prepared using distilled water adjusted to pH 4.5 was added to 2.5 g of dried and ground pepper leaves, followed by extraction at 180 rpm at room temperature for 3 hours. 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 ultrahigh pressure homogenized pepper leaf extract. The extract was used in Experimental Example 1.

(2) Preparation of Red Pepper Leaf Ethanol Extract

100 ml of 70% (v / v) ethanol prepared by distilled water adjusted to pH 4.5 was added to 2.5 g of dried and ground red pepper leaves, followed by extraction at 180 rpm for 2 hours at room temperature, followed by 15 minutes at 12,000 rpm. After centrifugation and filtering, pepper leaf ethanol extract was prepared, which was used as Comparative Example 1 of the present invention.

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

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

(4) Preparation of Pectinex Enzyme Treatment and Ultra High Pressure Homogenization Parallel Treatment of Red Pepper Leaf Extract

2.5 g of dried and ground pepper leaves were added with 30 ml of distilled water adjusted to pH 4.5 and 500 units / ml of Pectinex, and stirred at 180 rpm for 2 hours at 37 ° C. Thereafter, 70 ml of 100% (v / v) ethanol was added thereto, followed by stirring extraction at 180 rpm for 2 hours at room temperature, followed by centrifugation at 12,000 rpm for 15 minutes, followed by filtering to prepare a filtrate, and then, under a pressure of 110 MPa. Two orifices were passed through the tube to prepare Pectinex enzyme treatment and ultra high pressure homogenization in parallel pepper leaf extract. The extract was used as Comparative Example 3.

EXAMPLE  1. Comparison of Yield of Red Pepper Leaves by Different Extraction Methods

The extraction yield of the red pepper leaf extract extracted for each condition in Preparation Example 1 was compared. As a result, as shown in Table 1, the extraction yield was increased when the enzyme treatment, ultrahigh pressure homogenization or enzyme treatment and ultrahigh pressure homogenization were compared with those of Comparative Example 1, which is the pepper leaf ethanol extract. Extraction yield was calculated by the following formula 1, the increase rate refers to the degree of increase based on Comparative Example 1.

[Equation 1]

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

Treatment group Extraction yield (%) % Increase Comparative Example 1
(Ethanol extraction) 31.60 - Comparative Example 2
(Pectex processing) 50.58 60.06 Comparative Example 3
(Ultra High Pressure Homogenization + Pectex) 51.15 61.87 Experimental Example 1
(Ultra High Pressure Homogenization) 37.60 18.99

EXAMPLE  2. Changes in Polyphenol Contents of Red Pepper Leaf Extracts According to Extraction Methods

The total polyphenol content of the pepper leaf extract extracted for each condition in Preparation Example 1 was confirmed. As a result, as shown in Table 2, when the enzyme treatment, ultra-high pressure homogenization or the enzyme treatment and ultra-high pressure homogenization were combined, it was confirmed that the total polyphenol content was increased compared to the pepper ethanol extract of Comparative Example 1.

Treatment group Polyphenol Content (mg / extract 1g) Comparative Example 1
(Ethanol extraction) 36.50 Comparative Example 2
(Pectex processing) 40.39 Comparative Example 3
(Ultra High Pressure Homogenization + Pectex) 40.10 Experimental Example 1
(Ultra High Pressure Homogenization) 37.61

EXAMPLE  3. Cell survival rate ( cell viability ) Measure

In order to confirm the effect of red pepper leaf extract extracted under each condition in Preparation Example 1 on the cell survival, MTT (3- (4,5-dimeth-ylthiazol-2-yl) -2,5 using Mosmann's (1983) method -diphenyltetrazolium bromide) analysis was performed. Adipocytes 3T3-L1 were incubated for one day in a 96-well plate containing 100 μl of DMEM medium, and then cultured for 24 hours by treating with 320ppm pepper leaf extract. Then, 10 μl of 5 mg / ml MTT solution was dispensed into each well, followed by incubation for 3 hours to induce a reduction reaction, and 100 μl of DMSO (dimethyl sulfoxide) solution was added to formazan in purple. ) The crystals were completely dissolved. Thereafter, the absorbance was measured at 570 nm using a spectrophotometer, and the relative cell viability of the pepper leaf extract treatment group was calculated based on the survival rate of 100% of the pepper leaf extract untreated group.

As a result, as shown in Figure 1, the pepper leaf extract of each condition of Preparation Example 1 was found to have little cytotoxicity, as it did not significantly affect the cell viability.

EXAMPLE  4. Inhibitory Effects of Red Pepper Leaf Extracts on Fat Production

Adipogenesis of 3T3-L1, adipose precursor cells, was measured by staining using Lipid (Oil Red-O) staining kit (sigma, USA). Prepared and differentiated by treating the pepper leaf extract of Preparation Example 320ppm and differentiation inducer (MDI), then treated with phosphate buffer saline (PBS) containing 10% formalin, fixed at room temperature, and then removed, 60 Washed with% isopropanol and dried. Then, Oil Red-O was added, dyed for 45 minutes, washed four times with distilled water, and distilled water was completely removed. Thereafter, 100% isopropanol was added to the cells, and Oil Red-O stained with fat for 10 minutes was extracted at 180 rpm, and the value thereof was measured using 520 nm absorbance.

As a result, as shown in Figure 2, the normal ethanol extract (Comparative Example 1) and the enzyme alone treatment group (Comparative Example 2) did not show the effect of inhibiting fat production, but the ultra-high pressure homogeneous treatment group (Experimental Example 1) has a It was inhibited by about 80%, and when combined with pectinex treatment and ultra-high pressure homogenization (Comparative Example 3), it was confirmed that the effect of inhibiting fat production did not appear.

Claims (6)

Health functional food composition for the prevention or improvement of obesity containing red pepper ethanol extract prepared by the method comprising the following steps as an active ingredient:
(1) drying the red pepper leaves and then grinding to prepare red pepper leaf powder;
(2) adding ethanol to the pepper leaf powder prepared in step (1), followed by stirring extraction;
(3) preparing a filtrate by centrifuging and filtering the ethanol extract of step (2); And
(4) after step (3), ultra-high pressure homogenizing the filtrate. delete According to claim 1, wherein the ultra-high pressure homogenization of step (4) is a health functional food composition for the prevention or improvement of obesity, characterized in that carried out under a pressure of 100 ~ 120MPa. According to claim 1, Ultra-high pressure homogenization of the step (4) is a health functional food composition for the prevention or improvement of obesity, characterized in that repeated 1 to 3 times. The health functional food composition for preventing or improving obesity, characterized in that the red pepper leaf ethanol extract inhibits fat accumulation. Pharmaceutical composition for the prevention or treatment of obesity containing red pepper ethanol extract prepared by the method comprising the following steps as an active ingredient:
(1) drying the red pepper leaves and then grinding to prepare red pepper leaf powder;
(2) adding ethanol to the pepper leaf powder prepared in step (1), followed by stirring extraction;
(3) preparing a filtrate by centrifuging and filtering the ethanol extract of step (2); And
(4) after step (3), ultra-high pressure homogenizing the filtrate.

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