WO2012077975A2

WO2012077975A2 - Anti-aging composition containing velvet apple extracts or fractions thereof as active ingredients

Info

Publication number: WO2012077975A2
Application number: PCT/KR2011/009429
Authority: WO
Inventors: 이형규; 안경섭; 권옥경; 박지원; 오세량; 정혁
Original assignee: 한국생명공학연구원
Priority date: 2010-12-07
Filing date: 2011-12-07
Publication date: 2012-06-14
Also published as: WO2012077975A3

Abstract

【Abstract】 The present invention relates to an anti-aging and aging retarding composition which contains velvet apple extracts or fractions thereof as active ingredients. More particularly, the velvet apple extracts of the present invention exhibit DPPH free radical scavenging effects and ABTS free radical scavenging effects in a dose-dependent manner. Particularly, ethyl acetate of velvet apple and butanol fractions thereof exhibit excellent antioxidant effects, and therefore, the velvet apple extracts of the present invention may be valuably used as active ingredients of an anti-aging and aging retarding composition, a pharmaceutical composition and a composition for health food.

Description

【Specification】

[Name of invention]

Anti-aging composition containing velvet apple extract or fractions thereof as an active ingredient

The present invention relates to an anti-aging composition, specifically, anti-aging and delaying cosmetic composition, pharmaceutical composition and health food containing velvet apple extract (· ο> το5 blancoi A. DC.) Or fractions thereof as an active ingredient. To a composition.

Background Art

Free radicals, which cause aging and disease in the human body, are formed as biological reactions during normal metabolic processes in the human body, and are known to cause various diseases by causing harmful toxicity to cells and tissues (Folin 0 & Denis W, 1912, J Biol Chem 12: 239-249). The physiological action of inhibiting such harmful free radicals is a superoxide anion with a very similar reaction and destructive ability as it has a role similar to a superoxide dismutase (SOD) and an electron donating ability that inhibits oxidation by donating electrons to oxidative free radicals. It is responsible for converting radicals (superoxide anion radicals) into normal oxygen. Many bioactive compounds with strong antioxidant effects contribute greatly to the prevention of ulcers and spasms caused by oxidative stress, control of gastric juice secretion and diarrhea, prevention of diabetes, prevention of cancer, heart disease and degenerative diseases. (Azuma K et al., 1999, J Agric Food Chem 47: 3963-3966, Ham SS et al., 1997, J Food Sci Nutr 2: 155-161). In recent years, with increasing interest in health, we have selected resources containing a large amount of functional substances, such as anti-cancer, anti-allergic, anti-obesity, antioxidant, and antibacterial, from various plant sources, as drugs, food additives, or cosmetics. I want to develop from raw materials Globally (Shin DB, et al., 2006, Food Sci Biotechnol., 15: 357-362, Park et al., 2004, Korean J Nutr, 37: 364-372, Kim et al., 2006, Kor J Food Sci.Techno 1. 38: 799-804, Ali KA et al., 2005, Food Chem., 89: 27-36) Superoxide dismutase, an antioxidant, is a free radical produced by the respiratory process of cells. Decomposes superoxide radicals into hydrogen peroxide and oxygen. When hydrogen peroxide reacts with metal ions such as iron or copper ions, it generates more powerful active oxygen called hydroxy radicals. The antioxidants catalase and glutathione peroxidase, which remove hydrogen peroxide before it reacts with metal ions, decompose hydrogen peroxide into water and carbon dioxide. The antioxidant that removes hydrogen peroxide does not completely remove 5% of hydrogen peroxide, and it reacts with metal silver in the body, turning it into hydroxy radicals, aging our bodies and causing them to die. The found exposure of radiation or ultraviolet light converts hydrogen peroxide into hydroxy radicals, which promotes aging, and the method of delaying this is the use of antioxidants. Superoxide radicals (0 ^{2 "} ) themselves are relatively weak in reaction, but are easily converted to ¾0 ₂ and eventually produce highly reactive hydroxyl radicals or react with nitric oxides (NO) It produces strong peroxy nitrite (0N00), which causes oxidation of SH-groups, nitrification of protein tyrosine, lipid peroxidation, DNA damage, etc. Antioxidants are widely used in the plant system. It is distributed in many fruits and vegetables, including phenolic compounds, flavonoids, tocopheres, antioxidants such as vitamin C and selenium, which delay or prevent the oxidation of fat and prevent and delay cancer and cardiovascular diseases. Tocopherols and vitamin C, which are natural antioxidants in the plant kingdom, are also used in food, medicine and cosmetics. Li is used. Oxidation of maintenance or maintenance-containing food is ileonaneunde primarily by combining with oxygen in the air, In order to prevent this, many synthetic or natural antioxidants have been developed. However, due to its effectiveness and economical efficiency, synthetic antioxidants are actually used a lot. Synthetic antioxidants include BHT (Butylated Hydroxy Toluene), BHA (Butylated Hydroxy Anisole), Propyl Gal late (Propyl Gal late). Synthetic antioxidants are widely used in commercial foods because of their excellent antioxidant power and low price, but their usage is legally regulated due to concerns about stability such as human side effects. Natural antioxidants, such as tocopherol, are safe for the human body but have the ability to block oxidative chain reactions alone, and are expensive. Therefore, in recent years, research has been actively conducted to develop safer and more natural antioxidants. Velvet apple belongs to the Ebenaceae family, is called Mabolo or Kagong, and is a plant of the genus hawthorn. It is native to the Philippines, and the velvet apple tree is colored in quality, and its structure is hard, and it is also called “iron-wood.” The fruit of the velvet apple is surrounded by velvety reddish brown shell, and its taste and aroma are fruit cream. It is similar to cheese (Benedikt Mandl, Jimmy Wales, Wikimedia Foundation, 2004) .U.S. Patent 20080199533 has been reported to be used as an ingredient in skin improvement cosmetics. The antioxidant efficacy of the extracts has not been reported, and thus, the inventors of the present invention screened plants having excellent antioxidant effects, while the velvet apple extract was effective in eliminating DPPH (l, l-diphenyl_2-picryl—hydrazyl) free radicals. (2, 2 '-az i nob i s- (3-et hy 1 benzo thi azo 1 i ne-6-su 1 phon ic acid)) Free radical scavenging effect and superoxide radical scavenging effect Since the velvet apple ethyl acetate and butanol dissolving fractions exhibited excellent radical scavenging effect, the present invention was completed by revealing that the velvet apple extract or its fractions could be usefully used as an active ingredient for anti-aging and delaying compositions. It was. [Detailed Description of the Invention]

[Technical problem]

The object of the present invention is a velvet apple

blancoi A. DC.) To provide an anti-aging and delayed cosmetic composition, pharmaceutical composition and health food composition containing the extract or fraction thereof as an active ingredient.

Technical Solution

In order to achieve the above object, the present invention provides a cosmetic composition for anti-aging and delaying containing velvet apple ( ^' ο το5 blancoi A. DC.) Extract or fraction thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for anti-aging and delaying containing the velvet apple extract or a fraction thereof as an active ingredient.

In addition, the present invention provides a composition for anti-aging and delaying health food containing velvet apple extract or a fraction thereof as an active ingredient.

The present invention also provides a method for preventing and delaying aging comprising administering a pharmaceutically effective amount of a velvet apple extract or a fraction thereof to a subject. The present invention also provides a velvet apple extract or a fraction thereof for use as an anti-aging and delaying cosmetic composition.

The present invention also provides a velvet apple extract or a fraction thereof for use as a pharmaceutical composition for anti-aging and delaying.

In addition, the present invention provides a velvet apple extract or fractions thereof for use as an anti-aging and delaying health food composition. Hereinafter, the present invention will be described in detail. The present invention provides a cosmetic composition for anti-aging and delaying containing velvet apple ( ^' ο το5 blancoi A. DC.) Extract or fractions thereof as an active ingredient. The aging is preferably skin aging but is not limited thereto.

The present invention also provides a cosmetic composition for antioxidant containing velvet apple extract or a fraction thereof as an active ingredient.

The velvet apple extract is preferably prepared by a manufacturing method comprising the following steps, but not always limited thereto:

1) Velvet Apple

blancoi A. DC.) extract by adding an extraction solvent;

2) filtering the extract of step 1);

3) concentration of the filtered extract of step 2) under reduced pressure and drying to prepare an extract of velvet apple; And

4) velvet apple extractol of step 3) additionally extracting with an organic solvent to prepare a velvet apple fraction.

In the above method, the velvet apple bJancoi ^' A. DC.) Of step 1) can be used without limitation, such as grown or commercially available. The velvet apple is a leaf, beetle or root all available, it is most preferably a leaf, but is not limited thereto.

In the above method, the extraction solvent of step 1) is preferably water, alcohol or a mixture thereof. As the alcohol, it is preferable to use d to c ₂ lower alcohol, and it is preferable to use ethanol or methanol as the lower alcohol. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but is not limited thereto. The extraction solvent is preferably extracted by adding 1 to 10 times the amount of dried velvet apple, and more preferably by adding 2 to 3 times the extraction solvent. The extraction temperature is preferably 20 ° C to 100 ° C, more preferably 20 ° C to 40 ° C, most preferably room temperature, but is not limited thereto. In addition, the extraction time is preferably 10 to 48 hours, more preferably 15 to 30 hours, most preferably 24 hours, but is not limited thereto. In addition, the number of extraction is preferably 1 to 5 times, repeated extraction 3 to 4 times It is more preferable, and it is most preferable that it is three times, but it is not limited to this. In the above method, the decompression concentration in step 3) preferably uses a vacuum decompression concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably dried under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying, but not always limited thereto.

In the above method, the organic solvent of step 4) is preferably, but not limited to, normal-nucleic acid, chloroform, ethyl acetate or butanol. The fraction is obtained by suspending the velvet apple extract in water and then systematically fractionating the mixture into normal-nucleic acid, chloroform, ethyl acetate, normal-butanol, and water. Or it is preferably any one of the water fraction, and more preferably, ethyl acetate fraction, but is not limited thereto. The fraction may be obtained by repeating the fractionation process from 1 to 5 times, preferably 3 times from the velvet apple extract, and preferably concentrated under reduced pressure after the fraction, but is not limited thereto.

In order to confirm the antioxidant effect of the velvet apple extract, the present inventors investigated the DPPH free radical scavenging effect of 1,1-diphenyl-2-picrylylhydrazyl (l, l-diphenyl-2-picryl ᅳ hydrazyl, DPPH, Sigma). ), The velvet apple extract showed significant DPPH free radical scavenging effects of 18 ᅳ 32, 49, 68, 79 and 84% at 2.5, 5, 10, 20, 40 and 80 ^ g / mi concentrations, respectively. It was confirmed (see FIG. 1).

In addition, the present inventors measured the ABTS free radical scavenging effect according to various concentrations of the velvet apple extract, in order to confirm the antioxidant effect of the velvet apple extract, each velvet apple extract 2.5, 5, 10, 20, 40 and 80 It was confirmed that the g / nrf concentration showed a significant ABTS free radical scavenging effect of 17, 25, 39, 62, 83 and 97%, respectively (see FIG. 2).

In addition, the present inventors measured the superoxide radical scavenging effect of each concentration of the velvet apple extract using the SOD kit to confirm the SOE Superoxide Di smut ase) activity of the velvet apple extract, Velvet apple extract 2.5, 5, 10 ， 20 ， 40 and 80 It was confirmed that a significant superoxide radical scavenging effect of 12, 16, 24, 37 and 46% at / ig / i concentration, respectively (see FIG. 3).

In addition, the present inventors measured the DPPH and ABTS free radical scavenging effect in order to confirm the antioxidant effect of the velvet apple fraction, the velvet apple pole fraction was found to have excellent DPPH and ABTS free radical scavenging effect, in particular velvet apple ethyl acetate Fractions and butane fractions were found to have a significantly higher DPPH and ABTS free radical scavenging effect. In addition, when compared with vitamin C, a positive control group derived from natural products, it was confirmed that the chloroform layer, ethyl acetate layer, butanol layer fractions showed significantly superior activeols (see Table 1).

Therefore, the velvet apple extract showed excellent DPPH free radical scavenging effect, ABTS free radical scavenging effect and superoxide radical scavenging effect, and the velvet apple dissolving fraction showed excellent radical scavenging effect. And it can be seen that it can be usefully used as an active ingredient of the delayed cosmetic composition. The cosmetic composition may be, for example, an emulsion, suspension, microemulsion, microcapsules, microgranules or ionic (liposomes), nonionic, obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, or water phase. It may be provided in the form of a vesicle dispersant, cream, skin, lotion, powder, ointment, spray or cone stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition is in addition to the velvet apple extract or fractions thereof of the present invention, fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, fragrances , Surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics Cosmetics such as any other ingredient commonly used in It may contain adjuvants commonly used in the field of PT / KR2011 / 009429.

In the cosmetic composition containing the velvet apple extract of the present invention or a fraction thereof, the extract or fraction of the present invention is added to the cosmetic composition usually contained in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight. Can be.

When the velvet apple extract of the present invention or a fraction thereof is used as an external preparation for skin, in addition to fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, Fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients commonly used in external preparations for the skin. In addition, the ingredients can be introduced in amounts generally used in the field of dermatology.

The aging is preferably skin aging but is not limited thereto.

The present invention also provides a pharmaceutical composition for antioxidant containing velvet apple extract or a fraction thereof as an active ingredient.

The velvet apple extract of the present invention exhibited excellent DPPH free radical scavenging effect, ABTS free radical scavenging effect, and superoxide radical scavenging effect. It can be seen that it can be usefully used as an active ingredient of the preventive pharmaceutical composition. The composition containing the velvet apple extract or fractions thereof of the present invention may further contain one or more active ingredients exhibiting the same or similar functions in addition to the above components. have.

The composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate. , Lactose, Manni, Peel, Arabian rubber Pregelatinized starch, Corn starch, Powdered salose, Hydroxypropyl cellulose, Opadry, Sodium starch glycolate, Carnauba lead, Synthetic aluminum silicate, Stearic acid, Magnesium stearate ， Aluminum stearate, calcium stearate, sucrose, dextrose, sorbitan and talc may be used. The pharmaceutically acceptable additive according to the present invention is preferably included in the composition of 0.1 to 90 parts by weight, but is not limited thereto. It doesn't happen.

In other words, the composition of the present invention can be administered in various oral and parenteral dosage forms in actual clinical administration, and when formulated, diluents such as layering agents, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. that are commonly used, or It can be formulated using excipients. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like. Such solid preparations include at least one excipient such as starch, Calcium carbonate, velvet apple extract or fractions thereof. It can be prepared by mixing sucrose, lactose or gelatin. In addition to simple excipients, lubricants such as magnesium styrate talc may also be used. Oral liquid preparations include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. Non-aqueous solvents and suspending solvents may include propylene glycol, polyethylene glycol, vegetable oils such as lyb oil, and injectable esters such as ethyl acrylate. Suppository bases include witepsol, Macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like can be used. The composition of the present invention can be administered orally or parenterally according to the desired method, and when parenteral administration, external skin or intraperitoneal injection, rectal injection subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method It is desirable to choose. Dosage ranges depending on the patient's weight, age, sex, health status, diet administration time, administration method, excretion rate and the severity of the disease.

The dosage of the composition of the present invention varies depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate and severity of the disease, the daily dosage is the velvet Apollo extract or its 0.0001 to 100 mg / kg, preferably 0.001 to 10 rag / kg, based on the amount of fraction, and may be administered 1 to 6 times per day. The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological reaction modifiers.

The present invention also provides a composition for preventing and delaying health food containing velvet apple extract or fractions thereof as an active ingredient.

The aging is preferably skin aging but is not limited thereto.

The present invention also provides an antioxidant health food containing the velvet apple extract or a fraction thereof as an active ingredient.

Velvet apple extract showed excellent DPPH free radical scavenging effect, ABTS free radical scavenging effect, and superoxide radical scavenging effect, and velvet apple pole dissolving fraction showed excellent radical scavenging effect. It can be seen that it can be usefully used as a positive component of.

The health food of the present invention may be added as it is, velvet apple extract or fractions thereof, or may be used with other foods or food ingredients, and may be suitably used according to a conventional method.

There is no particular limitation on the kind of the health food. Examples of foods to which the velvet apple extract or fractions thereof 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 and vitamin complexes, and includes all healthy foods in the ordinary sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01-0.04 g, preferably about 0.02-0.03 g per 100 m of the composition of the present invention.

In addition to the above, the health food of the present invention includes various nutrients, 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. And carbonation agents used in carbonated beverages. Others may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention. In addition, the present invention provides an anti-aging and delaying method comprising administering a pharmaceutically effective amount of a velvet apple extract or a fraction thereof to a subject. The aging is preferably skin aging, but is not limited thereto.

The pharmaceutically effective amount is 0.00001 to 10 mg / kg, preferably 0.0001 to 1 mg / kg, but is not limited thereto. Dosage may vary depending on the weight, age, gender, health, diet, duration of administration, rate of removal, severity of the disease, and the like of a particular patient. The subject is a vertebrate and preferably a mammal, more preferably an experimental animal such as a rat, rabbit, guinea peak, hamster, dog or cat, and most preferably an ape-like animal such as a chimpanzee or gorilla.

The administration method is preferably oral or parenteral, the skin application is most preferred. The present invention also provides a velvet apple extract or fractions thereof for use as an anti-aging and delaying cosmetic composition.

In addition, the present invention provides a velvet apple extract or fractions thereof for use as an anti-aging and delaying health food composition.

The velvet apple extract of the present invention exhibited excellent DPPH free radical scavenging effect, ABTS free radical scavenging effect and superoxide radical scavenging effect, and the velvet apple dissolving fraction showed excellent radical scavenging effect, thus aging the velvet apple extract or fractions thereof. It can be seen that it can be usefully used as an active ingredient of the preventive and delayed cosmetic compositions, pharmaceutical compositions and compositions for health food.

Advantageous Effects

Velvet Apple? / O¾Dyros blancoi A. DC.) Extract or fractions thereof exhibits excellent antioxidant effects due to concentration-dependent inhibition of DPPH free radical production, ABTS free radical production and superoxide radical inhibition. By confirming, it can be usefully used as an active ingredient of anti-aging and delaying cosmetic compositions, pharmaceutical compositions and health food compositions.

[Brief Description of Drawings]

1 Free radical scavenging of velvet apple extract Diospyros blancoi A. DC. The graph shows the effect.

Figure 2 is a graph showing the ABTS scavenging effect of the velvet apple extract.

Figure 3 is a graph showing the effect of superoxide dismutase (SOD) activity of the velvet apple extract.

Figure 4 is a graph showing a method for producing a solvent-specific fractions in Bellbat Apple methanol extract.

[Best form for implementation of the invention]

Hereinafter, the present invention will be described in detail by Examples, Experimental Examples and Preparation Examples.

However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following Examples, Experimental Examples, and Preparation Examples.

Example 1 Preparation of Velvet Apple Extract

The present inventors purchased the velvet apple (Z? / O¾ TOS blancoi A. DC.) Extract from the Overseas Biomaterials Herb Center located in Korea Research Institute of Bioscience and Biotechnology. Looking at the extraction process, 10 kg of the leaves were dried and pulverized, and then powdered, 20 ¬ of methanol was added to the velvet apple leaves, and the filtration supernatant was recovered by extracting for 24 hours while circulating at room temperature. This process was repeated three times to collect the supernatant, and then concentrated under reduced pressure to obtain 1.2 kg of velvet apple methanol extract.

Example 2 Preparation of Velvet Apple Solvent Fraction

In 1,068 mg of the velvet apple methanol extract obtained in <Example 1> was suspended by addition of water, and the mixture was shaken for 3 times to obtain the nucleic acid fraction 363.5. Then, the nucleic acid fractions were removed, and the same amount of chloroform was added to the remaining water layer. The mixture was left to shake three times to obtain 195.9 mg of the chloroform fraction. The same amount of ethyl acetate was added to the remaining water layer to obtain 97.8 mg of ethyl acetate fraction in the same manner. Obtained and left over The same amount of butane was added to TKR2011 / 009429 water buffalo to obtain 199.9 mg of butanol fraction in the same manner, and the remaining water layer was concentrated to obtain 130.3 mg of water fraction (FIG. 4).

Experimental Example 1 Confirmation of DPPH Free Radical Scavenging Effect of Velvet Apple Extract

The free radical scavenging effect of the velvet apple extract obtained in Example 1 was 1,1-diphenyl-2-picryl-hydrazil (1,1-131 ^ 1-2- ^ 1-1 ^^ 32 1, DPPH, Sigma) was measured. The free radical scavenging effect was measured by placing 90 {Λ of 0.15 mM DPPH solution dissolved in ethanol in a 96-well plate, and dispensing the extract at concentrations of 2.5, 5, 10, 20, 40 and 80 / g / urf, respectively. After standing at room temperature for 20 minutes, the absorbance was measured at 517 nm with an ELISA reader. The free radical scavenging effect ^) was calculated by the following equation. At this time, the group treated with DMS0 was used as the negative control group, and the group treated with the synthetic antioxidant BHA was used as the positive control group.

Equation 1 free radical scavenging activity (%) 二

^χ ΐοο

A: absorbance of the negative control group not treated with the velvet apple extract of the present invention; And B: absorbance of the experimental group treated with velvet apple extract of the present invention. As a result, as shown in FIG. 1, DPPH free radical scavenging effect of the velvet apple extract was measured, and each of the velvet apple extracts 2.5, 5, 10, 20, 40, and 80 iLglxwi concentrations was 18, 32, and 49 , 68, 79 and 84% of the DPPH free radical scavenging effect was shown, it was confirmed that the activity similar to the positive control BHA (Fig. 1). Experimental Example 2 Confirmation of ABTS Free Radical Scavenging Effect of Velvet Apple Extract The velvet apple extract obtained in <Example 1> was used to confirm the ABTS free radical scavenging effect. ABTS free radical scavenging effect was mixed with ABTS and potassium persulfate to a final concentration of 7 mM and 2.45 mM, respectively, and left for 12 hours in the dark to form ABTS radicals. It was distilled with distilled water so that the absorbance was 0.7 at 734 nm, and the plate was divided into 96 well plates, and the sample was dispensed at a concentration of 2.5, 5, 10, 20, 40, or 80 ^ ag / iii, and then stored at room temperature. The reaction was carried out in the dark for 7 minutes. Absorbance was measured using a microplate meter at 734 nm. At this time, the group treated with DMS0 was used as the negative control group, and the group treated with the synthetic antioxidant BHA was used as the positive control group.

As a result, as shown in Figure 2 measured the ABTS free radical scavenging effect on the velvet apple extract, velvet apple extract 2.5, 5, 10, 20, 40 and 80 jug / n concentrations of 17, 25, 39 respectively , 62, 83, 97% showed a significant ABTS free radical scavenging effect (Fig. 2). Experimental Example 3 SOD Activity of Velvet Apple Extract

In order to confirm the antioxidant effect of the extract of Example 1, the scavenging effect of the superoxide radical acting as a precursor of other harmful active oxygen species was measured. The superoxide radical scavenging effect can be detected by measuring the activity of the superoxide dismutase (SOD), an enzyme that eliminates superoxide by dismutating the superoxide. In the present invention, the kit S0D (kitXDojindo ) Was used. Specifically, a sample of each concentration was dispensed into a 96-well plate, and a water-soluble WST-1 (2- (4- indophenyl) -3- (4-nitrophenyl) -5 ᅳ (2,4) reacted with a superoxide radical. After the addition of disulfophenyl) -2H-tetrazolium), the SOD enzyme solution was added and reacted at 37 ° C. for 20 minutes, and the absorbance of the reaction solution was measured at 450 nm using a microplate meter. The superoxide radical scavenging effect of each sample was calculated by reducing reduction of WST relative to the control group, and the concentration of the sample which scavenged the superoxide radical 50% was set as IC50. At this time, DMS0 The group was used as a negative control group and the group treated with the synthetic antioxidant BHA was used as a positive control group.

As a result, as shown in Figure 3, the result of measuring the superoxide radical scavenging effect on the velvet apple extract, each velvet apple extract 2.5, 5, 10, 20, 40 and 80

The superoxide radical scavenging effects of 12, 16, 24, 37 and 46% were also observed at the concentrations, and the positive control group ΒΗΑ inhibited 20.5 土 1.03% at a concentration of 10 ug / ml. 23.4 士 2.36% was confirmed to inhibit the same concentration was confirmed that the activity is excellent (Fig. 3).

Experimental Example 4 DPPH and ABTS Free Radical Scavenging Ability of the Velvet Apricot Fraction

For the nucleic acid for each solvent fractionated in <Example 1>, chloroform, ethyl acetate, butanol, and water fractions, DPPH and the same method as in <Experimental Example 1> and <Experimental Example 2>

ABTS free radical scavenging ability was measured. Each fraction was dispensed at a concentration of 40 / g / m £, reacted at room temperature, and absorbance was measured with a microplate absorber. At this time, the group treated with DMS0 was used as a negative control group, and the group treated with vitamin C, a natural antioxidant, was used as a positive control group.

As a result, as shown in Table 1, it was confirmed that the velvet apple fraction was excellent in DPPH and ABTS free radical scavenging effect, and in particular, the velvet apple ethyl acetate fraction and butanol fraction were remarkably high in DPPH and ABTS free radical scavenging effect. . In addition, when compared with vitamin C, a positive control group derived from natural products, it was confirmed that the chloroform layer, ethyl acetate layer, butanol layer fractions showed significantly superior activity (Table 1). Table 1

Sample (40 // g / itrf) DPPH radical scavenging activity (%) ABTS radical scavenging activity (%)

Hll tfll ofl S. gll— tLsil S- 28.21 ± 0.07 28.63 ± 0.96 Velvet Apple

55.68 ± 0.71 63.35 ± 3.22

Chloroform fraction

Velvet Apple

89.18 ± 2.00 99.97 土 0.26

Ethyl acetate fraction

Velvet Apple Butanol Fraction 83.04 ± 1.42 99.81 ± 0.12

Velvet Apple Water Fraction 29.81 ± 0.88 45.47 ± 1.40

Vitamin C 37.14 ± 1.47 61.00 ± 3.61

Preparation Example 1 Preparation of Anti-aging Cosmetics Containing Velvet Apple Extract or Fractions thereof as an Active Ingredient

<1-1> Preparation of Flexible Lotion

The flexible lotion containing the velvet apple extract of <Example 1> or a fraction thereof as an active ingredient was prepared as shown in [Table 2].

Table 2

Raw Material 口 ᄋ ό ᄋ 丁) Velvet Apple Extract of the Invention

10.00

Or fractions thereof

1,3—butylene glycol 1.00

Dessert ID 0.05

Allantoin 0.10

Depotam glycylizate 0.05

Citrix Acid 0.01

Sodium Citrate 0.02

Glyceres -26 1.00

Arbutin 2.00 1 Hydrogenated castor oil 1.00 ethane 30.00

Preservative Trace

Trace amount of colorant

Trace amount of flavor

Purified water level

<1-2> Preparation of nourishing cream

The nutrition cream containing the velvet apple extract of <Example 1> or a fraction thereof as an active ingredient was prepared as shown in the following [Table 3].

[Table 3]

Raw material content (part by weight) Velvet Apple Extract of the present invention or

i

10.0

Fractions thereof

1,3-butylene glycol 7.0

Glycerin 1.0

D-panthenol 0.1

Botanical extracts 3.2

Magnesium Aluminum Silicate 0.3

PEG-40 Stearate 1.2

Stearic Acid 2.0

Polysorbate 60 1.5

Lipid Glyceryl Stearate 2.0

Sorbitan Sesquioleate 1.5 Cetearyl Alcohol 3.0

Mineral Oil 4.0

Squalane 3.8 Carlylic / Caprictriglycerides 2.8 Vegetable Oil 1.8

Dimethicone 0.4

Dipotassium glycylizate traces

Allantoin Trace

Sodium Hyaluronate Trace

Tocopheryl acetate appropriate amount

Triethanolamine appropriate amount

Preservative

Flavoring Agent

Purified water level

Preparation Example 2 Preparation of a Formulation Containing Velvet Apple Extract or Fractions thereof as an Active Ingredient

<2-1> Preparation of powder

Velvet apple extract of the present invention 2 g

1 g lactose

The above ingredients were mixed and layered in an airtight cloth to prepare a powder.

<2-2> Preparation of Tablet

100 nig of velvet apple ethyl acetate fraction of the present invention

Corn starch 100 mg

Lactose 100 mg Stearic Acid Magnesium 2 mg

The above-mentioned components were tableted-made by tableting according to the conventional manufacturing method of a common tablet.

<1-3> Preparation of the capsulant

100 mg of velvet apple extract of the present invention

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above ingredients was filled into gelatin capsules in accordance with the conventional method for producing a capsule to prepare a capsule.

<1-4> Preparation of the ring

Velvet apple extract of the present invention

ᄋ

1.5 g per ΤΓ

1 g of glycerin

0.5 g of Xili

After mixing the above components, it was prepared to be 4 8 per ring according to a conventional method.

<1-5> Preparation of Granules

Velvet Apple Extract 150 mg of the present invention

Soy extract 50 mg

Glucose 200 nig

Starch 600 nig

After mixing the above ingredients, 100 mg of 30% ethanol was added and dried at 60 ^° C. to form granules, and then filled into fabrics. Preparation Example 3 Preparation of Anti-Aging Health Food Containing Velvet Apple Extract or Fractions thereof as an Active Ingredient

<3-1> Preparation of Flour Food

0.5-5.0 parts by weight of the velvet apple extract or fractions thereof of the present invention was added to the flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture.

<3-2> Preparation of Soup and Gravy

0.1-5.0 parts by weight of the velvet Apollu extract or fractions thereof of the present invention was added to soups and gravy to prepare health products, noodle soup and gravy.

<3-3> Preparation of ground beef

10 parts by weight of the velvet apple extract or fractions thereof of the present invention was added to the ground beef to prepare a ground beef for health promotion.

<3-4> Manufacture of dairy products

5 to 10 parts by weight of the velvet apple extract or fractions thereof of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<3-5> Preparation of wire

Brown rice, barley, glutinous rice, and yulmu were roasted by alpha in a known method and then dried to prepare a powder having a particle size of 60 mesh.

Black beans, black sesame seeds, and sesame seeds were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh.

The velvet apple extract or fractions thereof of the present invention were concentrated under reduced pressure in a vacuum concentrator, dried by spraying and drying with a hot air dryer, and then pulverized with a particle size of 60 mesh to obtain a dry powder. The grains, seeds and the velvet apple extract of the present invention or fractions thereof were prepared by combining the following ratios.

Cereals (30 parts by weight brown rice, 15 parts by weight of radish, 20 parts by weight of barley) ，

Seeds (7 parts by weight perilla, 8 parts by weight black beans, 7 parts by weight black sesame),

Velvet apple extract or fractions thereof of the present invention (3 parts by weight),

Manor (0.5 parts by weight) ，

Foxglove (0.5 part by weight)

Preparation Example 4 Preparation of Anti-Aging Beverage Containing Velvet Apple Extract or Fractions thereof as an Active Ingredient

<4-1> Preparation of Health Beverage

Homogeneous blend of subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%) and the velvet apple extract of the present invention or fractions thereof After instant sterilization it was prepared by packaging in a small packaging container such as glass bottles, plastic bottles.

<4-2> Preparation of Vegetable Juice

Vegetable juice was prepared by adding 5 g of the velvet apple extract of the present invention or a fraction thereof to 1,000 tomato or carrot juices. <4-3> Preparation of fruit juice

Apple or grape juice of 1 g of the velvet apple extract or fractions thereof of the present invention

1,000 ml was added to prepare fruit juice.

Claims

[Range of request]

[Claim 11

Anti-aging and delaying cosmetic composition containing velvet apple ( ^' ο¾σ «blancox ^' A. DC.) Extract or fractions thereof as an active ingredient.

[Claim 2]

The anti-aging and delaying cosmetic composition according to claim 1, wherein the velvet apple extract is extracted using water, a lower alcohol of C ₄ or a mixture thereof as a solvent.

[Claim 3]

3. The anti-aging and delaying cosmetic composition according to claim 2, wherein the lower alcohol is ethane or methanol.

[Claim 4]

The method of claim 1, wherein the fraction is n-hexane, chloroform, ethyl acetate, _n- butanol water obtained by sequentially fractionating n-nucleic acid fraction, chloroform fraction, ethyl acetate fraction, n-butane fraction or water Anti-aging and delaying cosmetic composition, characterized in that the fraction.

[Claim 5]

Anti-aging and delaying pharmaceutical composition containing velvet apple extract or a fraction thereof as an active ingredient.

[Claim 6]

According to claim 5, The velvet apple extract is anti-aging and characterized in that the extraction using water, lower alcohol of C ₂ or a mixture thereof as a solvent Delayed pharmaceutical composition.

[Claim 71

7. The anti-aging and delaying pharmaceutical composition according to claim 6, wherein the lower alcohol is ethanol or methane.

[Claim 8]

The _n -nucleic acid fraction, chloroform fraction, ethyl acetate fraction, n—butanol fraction or water obtained by sequentially fractionating the velvet apple extract with n-nucleic acid, chloroform, ethyl acetate, _n -butanol and water. Anti-aging and delayed pharmaceutical composition, characterized in that the fraction.

[Claim 9]

Anti-aging and delaying health food composition containing velvet apple extract or fractions thereof as an active ingredient.

[Claim 10]

10. The anti-aging and delaying health food composition according to claim 9, wherein the velvet apple extract is extracted using water, ethanol or a mixture thereof as a solvent.

[Claim 11]

10. The method according to claim 9, wherein the fraction is _n -nucleic acid fraction, chloroform fraction, ethyl acetate fraction, n-butanol fraction obtained by sequentially fractionating velvet apple extract with n-nucleic acid, chloroform, ethyl acetate, _n- butane, and water or Anti-aging and delaying health food composition, characterized in that the water fraction.

[Claim 12]

A method of preventing and delaying aging comprising administering to a subject a pharmaceutically effective amount of a velvet apple extract or a fraction thereof.

[Claim 13]

Velvet apple extract or fractions thereof for use as anti-aging and delaying cosmetic composition.

[Claim 14]

Velvet apple extract or fractions thereof for use as a pharmaceutical composition for anti-aging and delaying.

[Claim 15]

Velvet apple extract or fractions thereof for use as an antiaging and delaying health food composition.