KR20190116837A - Composition Comprising Radish Extract for Improving Skin Wrinkle and Elasticity - Google Patents

Abstract

The present invention relates to a composition that is not irritating to the human body and is effective for reducing skin wrinkles and improving elasticity. More particularly, the present invention relates to a health functional food composition and cosmetic composition for reducing skin wrinkles and improving elasticity containing a radish extract.

Description

Composition Comprising Radish Extract for Improving Skin Wrinkle and Elasticity

The present invention relates to a composition that is not irritating to the human body and is effective for improving skin wrinkles and elasticity.

Wrinkles can be defined as overlapping skin phenomena caused by the loss of tension and elasticity of the skin. At first, the fine wrinkles are formed, but gradually become large and deep wrinkles. Due to the change in the characteristics and amount of extracellular matrix protein of the dermis, wrinkles are formed by changing physical properties including elasticity of skin or decrease of moisture and subcutaneous fat, and it starts to appear around 20 years old and gradually increases with age. The harder you use the exercise, the more wrinkles you get, so the face that uses a lot of muscles for facial expressions tends to wrinkle. Skin aging represented by wrinkles can be divided into two types.

The first is intrinsic aging, which is an unavoidable aging phenomenon over time. Clinical features are relatively mild and include fine lines, dry skin, and reduced elasticity.

The second is photoaging, which is the result of a combination of endogenous aging and the effects of UV light, which can be prevented by avoiding UV exposure. It is mainly observed on the skin of the face, back of the hand, back of the neck, etc., which has been exposed to the sun for a long time, and its clinical characteristics are more severe and observed earlier than endogenous aging. For example, irregular pigmentation occurs on the skin exposed to sunlight, pigment diseases such as sun light side increases, the skin becomes very rough and dry, elasticity decreases and the skin sags in severe cases. Wrinkles are also thicker and deeper than endogenous aging and have fine lines.

It is known that collagen synthesis decreases in the skin exposed to ultraviolet rays even if it is a small amount when walking outside in everyday life, and expression of matrix metalloproteinases (MMPs), which are collagen and extracellular matrix proteins, is increased. . The breakdown of matrix proteins by MMPs is a type of wound that the skin receives from the sun, and the wound healing process such as synthesizing new collagen continues, but the wound healing process is not always perfect. Aging of the skin, including wrinkles, is getting worse.

Due to the destruction of the ozone layer due to environmental pollution, the amount of ultraviolet rays reaching the ground is increased, and the area of ultraviolet rays is also changed to increase photoaging, and the risk is expected to increase further. Sunscreens are used to prevent the harmful effects of UV rays, but recently, sunscreens have been reported to have a possibility of skin cancer due to mutations. In addition, in case of sudden exposure to the sun, an appropriate amount of anti-irritants and antioxidants are used for soothing the skin. However, these methods have insufficient effects such as soothing or anti-inflammatory effects on the skin damaged from ultraviolet rays. Since skin damage caused by ultraviolet rays is ultimately associated with skin aging, a way to protect them is one of the cosmetic industry's concerns.

Therefore, researches to improve skin wrinkles and to promote elasticity have been conducted in various fields. For example, Patent No. 10-0454757 (Photoaging Anti-aging Agent and Skin Cosmetic) and Patent No. 10-1040507 (Containing Magnolia Egg Extract) Cosmetic composition for preventing photoaging and improving wrinkles of skin), Patent No. 10-0858630 (Cosmetic composition for preventing ultraviolet rays and photoaging containing mung bean extract), Patent No. 10-0537915 (including persimmon extract as an active ingredient) Cosmetic compositions for improving skin wrinkles and elasticity).

Radish (Raphanus sativus L.) is a cruciferae vegetable that has a volatile sulfur content and has a unique spicy taste. The unique spicy taste of radishes is due to the separation of thiocyanates and isothiocyanates by an enzyme called glucosidase in itself when the thioglucosides contained in the radish are cut off or the cells are destroyed. In Korea, with the introduction of Buddhism, it began to be cultivated in the Three Kingdoms period, and the largest cultivated area among Korean vegetables is 50,000 ha and the annual output reaches 2.2 million tons. Radish has a variety of uses because it tastes not only in side dishes such as kimchi, diced radish, radish, radish, and radish, but also in soup, stew, and stewed foods.

Radish contains more free amino acids, sugars, calcium and phosphorus than other vegetables. The sugars contained in the roots are mainly glucose and fructose, and also contain many organic acids and amino acids. In particular, the high content of vitamin C has been an important source of vitamins in winter since ancient times. Radishes, nabok (羅蔔) is said to be effective in sputum, coughing, dysentery, etc., and also in relieving poisoning of seafood or noodles. Diastase contained in radish is effective for promoting digestion, food poisoning, and hangover relief, and rapine is known to have antibacterial effects against bacteria, fungi and parasites.

Radish skin and radish are also used for food, but due to the tough and coarse texture, they are often eaten after removal, and are used as by-products or feed for livestock. The present inventors have completed the present invention while studying the processing method that can further increase the physiological activity of the radish can be useful for improving the wrinkles and elasticity of the processed radish including the by-products.

Patent Registration No. 10-0454757 Patent Registration No. 10-1040507 Patent Registration No. 10-0858630 Patent Registration No. 10-0537915

An object of the present invention is to provide a cosmetic composition that can be used safely without irritation or toxicity to the skin, while preventing wrinkle formation of the skin and improving skin wrinkles and elasticity.

In another aspect, the present invention to provide a dietary supplement composition effective for improving skin wrinkles and elasticity even when taken orally.

The present invention for achieving the above object relates to a cosmetic composition for improving skin wrinkles and elasticity containing a radish extract.

The radish extract was more preferably an extract of heat-treated radish, heat treatment of radish increased the yield of the extract, antioxidant properties and the inhibition of expression of MMPs was also improved (data not shown). The heat treatment is to heat the radish, it is preferable to process at a high temperature as possible, but it is preferable to heat treatment at 100 ~ 200 ℃ for 10 minutes to 6 hours, because the carbonization can occur when the temperature increases, 110 ~ 150 ℃ of More preferred was treatment at temperature. Naturally, the heat treatment time can be properly adjusted according to the size of the sample and the heat treatment temperature.

The radish may be root of radish or radish or mixtures thereof. Radishes and radishes are cleaned and used after removing contaminated parts. Whole radish and radish can be cut into whole or proper size. In the case of cutting to a small size, the useful materials may leak during the heat treatment, so it is good to treat them in containers so as to prevent the leakage of useful materials.

The radish may be used in any form, such as raw, dried or frozen products. In the case of using dried radish, it may be carbonized during the heat treatment during heat treatment at the temperature, so that it is preferable to heat the mixture by being called water or mixed. When the radish is heat-treated in a raw state or in a frozen state, carbonization does not occur because radish has its own moisture even without adding water. Heat treatment at the temperature in the presence of water, and in order to prevent the loss of useful components in the heat treatment process, it is preferable to heat treatment under high pressure conditions, radish is cooked in the heat treatment process.

 Extract of radish in the present invention can be obtained using the juice as it is, supercritical extraction, or using a separate solvent. In the case of extraction using a separate solvent, the solvent may be water or a lower alcohol of C1 ~ C4 or a mixture thereof. More preferably, the radish heat-treated during extraction with a solvent is extracted by grinding or grinding to facilitate extraction. Extraction may use any of the methods used for extraction of natural products in the art, for example, but may be used, such as cold extraction, heat extraction, ultrasonic extraction, but is not limited thereto. The extract obtained by extracting with juice or a solvent can be used by itself, concentrated or dried. It is obvious that the drying method may be a conventional method such as spray drying, heat drying, and freeze drying.

The composition of the present invention has the effect of inhibiting the expression of MMPs, which are enzymes of extracellular matrix proteins, inhibiting and improving the formation of wrinkles, and improving elasticity, and thus can be usefully used as a cosmetic composition. In particular, the composition of the present invention can be used for suppressing and improving skin wrinkles caused by photoaging caused by exposure to ultraviolet light.

In order to use the composition of the present invention as a cosmetic, it may further include components conventionally used in cosmetic compositions, for example, with conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances and Carrier and the like. Cosmetic composition of the present invention may contain an extract of radish in a proportion of 0.1 to 100% by weight. In addition to extracts of radish, it may further contain other useful ingredients.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, nutrition cream, massage cream, essence, eye cream, cleansing detergent, cleansing foam, mask pack, foundation, powder It may be prepared in the form of lotion, but is not limited thereto. The cosmetic composition of the present invention may be used alone or in duplicate, or may be used in combination with other cosmetic compositions other than the present invention. In addition, the cosmetic composition with excellent skin protection effect according to the present invention can be used according to a conventional method of use, the number of times of use can be varied according to the user's skin condition or taste.

The composition of the present invention is effective in improving skin wrinkles when applied to the skin as well as orally ingested, and can be usefully used as a health functional food composition. In particular, radish is a safe food that does not show any side effects even when taken for a long time can be obtained by improving the skin wrinkles and elasticity by taking it steadily for a long time. The food composition may be used in the form of tablets, capsules, pills or liquids as a health food, or may be used as additives of various foods, beverages, gums, teas, vitamin complexes. The extract of radish may be added to the food composition of the present invention preferably 0.01 to 100% by weight.

According to the composition containing the heat-treated radish extract of the present invention as described above, the biosafety is ensured when taken orally or applied to the skin, without side effects, to prevent and improve the skin wrinkles, and to improve the skin elasticity is excellent cosmetic It can be usefully used as a composition or nutraceutical composition.

1 is a graph showing the weight change of the experimental animal.
Figure 2 is a graph showing the dietary intake and dietary efficiency of the experimental animals.
Figure 3 is a graph showing the degree of wrinkles of experimental animals over time.
Figure 4 is a graph showing the expression level of skin wrinkles related genes of the experimental animal.
Figure 5 is a graph showing the expression level of skin wrinkles related proteins of the experimental animal.
Figure 6 is a photograph showing the stability of the cream according to an embodiment of the present invention.
Figure 7 is an image showing the elasticity measurement site for the clinical trial of the cream according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the accompanying examples. However, these embodiments are merely illustrative for easily describing the content and scope of the technical idea of the present invention, and thus the technical scope of the present invention is not limited or changed. It will be apparent to those skilled in the art that various modifications and variations are possible within the scope of the present invention based on these examples.

EXAMPLE

Example 1 Heat Treatment of Radish and Preparation of Extract

1) heat treatment of radish

Radishes discarded at the agricultural and fishery wholesale market were purchased and used after washing, including skin and radish.

The heat treatment apparatus was designed and manufactured to withstand the pressure of 10 kg / cm ² or more was used (Jisco, Seoul, Korea). The whole sample was placed in a container, and then placed in an external container to which a certain amount of water was added, and then heated according to a predetermined temperature and time to prevent carbonization of the sample by direct heat transfer. . The heat treatment temperatures were set to 110, 120, 130, 140 and 150 ° C., and the heat treatment time was set to 2 hours.

2) Preparation of Heat Treated Radish Extract (HRE)

After cooling the heat-treated sample, it was crushed using a grinder and extracted 10-14 times (v / v) of distilled water for 14-16 hours. The extract was filtered and then lyophilized.

The yield of ethanol extract was 5.709 w% when heat treated at 130 ° C. but decreased slightly to 5.584 and 5.186 w% at 140 and 150 ° C., respectively.

3) Preparation of heat treated supercritical extract (HO)

The heat-treated radish was dried in a well ventilated place for 24 hours and then ground to a size of 200 mesh or less. The pulverized product was placed in a supercritical fluid extractor, and supercritical CO ₂ was extracted using butylene glycol as an auxiliary solvent under the extraction temperature of 40-80 ° C. and the extraction pressure of 200-500 bar. The collected extract was likewise lyophilized and used.

Example 2 Animal Experiment

1) Preparation and treatment of experimental animals

Seven-week-old HRM-2 mice were fed and purified in the nursery for one week, and then used for the experiment. The experimental animals were kept at a temperature of 22 ° C. and a humidity of 50% while maintaining a 12-hour contrast cycle, and feed and water were freely fed.

After one week of purification, the sample prepared in Example 1 was orally administered once a day or the HRM-2 mouse was divided into two parts and the sample was applied to the right side. The effect of the sample on skin aging was evaluated by UVB irradiation. The experimental groups were classified as shown in Table 1 below, and the number of experimental animals per group was five.

UVB is used with a UVB lamp (15 W type, UV maximum wave length 312 nm; UV intensity 100 μW cm ^-2 , Ieda Boeki Co., Tokyo, Japan) at a luminous intensity of 100 mJ / cm ² daily for the first week. , From 2 weeks to 5 weeks, UVB was irradiated three times a week (Mon, Wed, Fri) at a distance of 15 cm at a brightness of 200 mJ / cm ² to induce skin aging. Table 2 below shows the experimental schedule.

2) weight and dietary intake

In order to determine the effect of treatment and UVB irradiation on the dietary intake and body weight of HRM-2 mice, body weight and dietary intake were measured and recorded at 10 pm every Thursday during the treatment of experimental animals of 1).

Figure 1 is a graph showing the change in weight over time, Figure 2 is a graph showing the dietary intake and dietary efficiency over five weeks. Dietary efficiency (FER%) is the ratio of weight gain in five weeks to dietary intake in five weeks.

1 shows that no significant weight change was observed between groups by irradiation of UVB or treatment of samples. In addition, there was no significant difference in dietary intake and dietary efficiency in the negative control group compared to normal group. The dietary intake of oral or oint-treated radish extracts was similar to that of the normal group, but the dietary efficiency was significantly decreased in all groups compared to the negative control group.

3) Effect of Radish Extracts on Wrinkle Formation by UVB Irradiation

To evaluate the effect of radish extract on wrinkle formation caused by UVB irradiation, skin dermobella 3D analyzer was used to measure the depth of skin wrinkles at 3, 4 and 5 weeks after the start of the experiment. Figure 3 is a graph showing the results, it can be observed that the depth of wrinkles of the negative control group compared to the normal group by UVB irradiation. On the other hand, the depth of wrinkles was reduced in the treatment group of radish extract compared to the negative control.

4) Evaluation of the Effect of Radish Extracts on the Expression of Wrinkle-related Genes and Proteins by UVB Irradiation

In the experiment of 3), it was confirmed that the radish extract has an effect of alleviating the wrinkle depth by photoaging, and thus quantitative evaluation was made by measuring the expression level of wrinkle-related genes and proteins by UVB irradiation.

For this purpose, the mice were sacrificed at the 5th week of UVB irradiation according to the experiment of 1), and then the skin tissues were separated by UVB irradiation. 500 μl of Trizol was added to the isolated back tissue, and the cells were pulverized with a homogenizer, and 100 μl of chloroform (CHCl ₃ ) was added thereto, followed by mixing for 15 seconds. After standing on ice for 15 minutes, the mixture was centrifuged at 13,000 rpm, and about 200 μl of the supernatant was collected, mixed with 200 μl of 2-propanol in the same amount, slowly shaken, and left for 15 minutes on ice. After centrifugation at 13,000 rpm again, washed with 80% EtOH and dried for 3 minutes in a vaccum pump to extract RNA.

3 μg of the extracted RNA was denatured at 75 ° C. for 10 minutes, followed by 2.5 μl of 10 mM dNTPs mix, 1 μl random sequence hexanucleotides (25 pmole / 25 μl), and 1 μl RNase inhibitor (20 U / μl) as RNA inhibitor. 1 μl 100 mM DTT, 4.5 μl 5 × RT buffer (250 mM Tris-HCl, pH 8.3, 375 mM KCl, 15 mM MgCl ₂ ) was added, followed by 1 μl of M-MLV RT (200 U / μl). It was added again and distilled water treated with DEPC (diethyl pyrocarbonate) to the final volume of 20 μl. 20 μl of the reaction mixture was mixed well and centrifuged at 2,000 rpm for 5 seconds to react for 60 minutes in a 37 ° C. heating block to synthesize first-strand cDNA, and then left at 95 ° C. for 5 minutes to burn M-MLV RT. Activated.

RT PCR was performed using the synthesized cDNA. Primer sequences used for PCR are described in Table 3 below, and the Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, USA) was used. TaqMan probe (FAM dye-labeled, ABi, USA) was used as an internal standard for mouse GAPDH probe set; Endogenous Control (VIC® / MGB Probe, Probe Limited, Applied Biosystems 4352339E) was used and reacted to a final concentration of 200 nM of primer. The conditions of real time quantitative PCR were pre-denaturation for 2 minutes at 50 ℃, 10 minutes at 94 ℃, and 40 cycles for 15 seconds at 95 ℃ and 1 minute at 60 ℃.

4 is a graph showing the results of RT PCR, UVB irradiation negative control group significantly increased the expression of MMP-2 and MMP-9 genes compared to the normal group, but the expression of the gene by the treatment of radish extract was significant It was confirmed that the enemy decreases. In addition, the expression level of the gene decreased depending on the throughput of the radish extract.

The expression level of MMP-2 protein from the skin tissues isolated from the above was measured by ELISA using the MMP-2 ELISA (R & D System, USA) kit for mice according to the manufacturer's manual. As shown in FIG. 5, the expression of MMP-2 protein was also greatly increased by irradiation of UVB, and the extract of radish inhibited the expression of MMP-2 protein depending on the throughput.

Example 3 Clinical Evaluation of a Cream Containing Radish Extract

Sweet almond oil and sunflower seed oil were mixed with 30 mL of each, heated to 80 ° C., and 80 mL of purified water was added thereto to emulsify the blender. As an antioxidant, 100 mg of tocopherol and 3% of the radish extract prepared in Example 2) were added to prepare a cream.

In order to confirm the stability of the cream during the experimental period of 30 days, the pH and degree of discoloration of the cream for 30 days were measured. The pH was almost unchanged from 5.67 on the first day to 5.70 after 30 days, and no discoloration was observed as shown in FIG. 6.

The safety of the cream was evaluated by patch test. Specifically, 1 mL of cream was applied to 1 cm 2 of the center of the back to check the skin reaction after 24 hours. As a result, no erythema, crusting and edema were observed, and it was concluded that the cream prepared therefrom was "non-irritant".

The number of subjects for clinical evaluation was calculated by G * Power 3.1 program, which consisted of 5 experimental groups and 5 control groups. For this purpose, ten women in their 40s who did not have a history of skin diseases such as atopic dermatitis, psoriasis and allergic reactions within the last 6 months were selected. The experimental period was designed for 4 weeks. After applying the cream with heat-free extract that was heat treated 7 times a week, the entire face was washed with the same cleansing product before the experimental treatment, 2 weeks and 4 weeks, and the elasticity was mexameter MX18 (CK electronics, Germany). The measurement site is shown in FIG.

Table 4 and Table 5 below show the results of measuring the elasticity of the experimental group and the control group, respectively. As the elasticity is closer to 1, the elasticity is higher, and the experimental group has an average increase in elasticity of 0.03386, and the control group has an average increase of 0.0255, indicating that the experimental group has a higher increase in elasticity than the control group.

<110> SEOWON UNIVERSITY INSTITUTE OF INDUSTRY-ACADEMY COLLABORATION <120> Composition Comprising Radish Extract for Improving Skin Wrinkle          and Elasticity <130> P0418-251 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 1 cagggaatga gtactgggtc tatt 24 <210> 2 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 actccagtta aaggcagcat ctac 24 <210> 3 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 aatctcttct agagactggg aaggag 26 <210> 4 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 agctgattga ctaaagtagc tgga 24

Claims (6)

Cosmetic composition for improving skin wrinkles and elasticity containing radish extract.
The method of claim 1,
The radish extract is a cosmetic composition for improving skin wrinkles and elasticity, characterized in that the extract of radish heat-treated at 100 ~ 200 ℃ for 10 minutes ~ 6 hours.
The method according to claim 1 or 2,
The radish is a cosmetic composition for improving skin wrinkles and elasticity, characterized in that the root or radish of radish or a mixture thereof.
The method according to claim 1 or 2,
The extract is extracted using water or alcohol of C1 ~ C4 or a mixture thereof, or a cosmetic composition for improving skin wrinkles and elasticity, characterized in that the supercritical extraction.
The method according to claim 1 or 2,
Cosmetic composition for improving skin wrinkles and elasticity by photoaging.
Health functional food composition for improving skin wrinkles and elasticity comprising an extract of radish.

Images