KR20190090242A - A composition improving skin wrinkle or striae distensae and having anti-inflammation effect comprising ailanthus altissima root bark extracts - Google Patents

Abstract

The present invention relates to a composition for reducing skin wrinkles, preventing and improving striae distensae, or having an anti-inflammatory effect, comprising an Ailanthus altissima root bark extract as an active ingredient. The composition comprising the Ailanthus altissima root bark extract of the present invention as an active ingredient is excellent in preventing and reducing skin wrinkles, improving skin color and skin texture of striae distensae, having an excellent anti-inflammatory effect, and having no cytotoxicity and skin side effects. In addition, the Ailanthus altissima root bark extract of the present invention can be safely used in cosmetics, pharmaceutical and food composition because there is no cytotoxicity and skin side effects.

Description

Technical Field [0001] The present invention relates to a composition for improving skin wrinkles, prevention and improvement of stretch marks, or a composition for anti-inflammation, which contains low molecular weight bark extract as an active ingredient, or a composition for anti-

The present invention relates to a low geunbaek blood extract on skin wrinkles comprising as an active ingredient, stretch marks prevention and improvement, or anti-inflammatory composition, more specifically, root bark of low geunbaek validate the blood extract in the skin tree (Ailanthus altissima) A cosmetic composition for preventing or treating skin wrinkles or stretch marks, a pharmaceutical composition, and a food composition.

As wellbeing and emotional consumption increase as the level of living improves and society ages, the value and aspiration for health and beauty grows and the appearance changes competitiveness, and healthy and beautiful skin is cultivated The desire of consumers to maintain is increasing. Therefore, consumers' interest and preference for functional cosmetics that emphasize efficacy and efficacy rather than merely beauty by using effective and stable natural materials in accordance with nature-friendly consumption tendency is increasing (Hyun et al., 2007). Natural products contain a large amount of useful physiologically active ingredients and have various pharmacological effects such as antioxidant, antibacterial, anti-inflammatory, anti-aging and anti-cancer activity. In particular, medicinal plants used as herbal medicinal herbs strengthen the body's immune system and help to maintain the body's homeostasis from external stresses, ultraviolet rays, environmental pollution and various chemicals including environmental hormones (Lee et al. 2000). Therefore, various studies on functional materials and products capable of delaying or inhibiting aging have been conducted (Kang et al., 2007; Jung et al., 2016). In recent years, physiologically active ingredients that can be applied to foods, cosmetics, medicines, and medicinal plants as a substitute for conventional synthetic chemical materials have been activated to minimize side effects and stimuli (Tsukahara et al , 2006).

Skin is the largest organ covering the external surface of human body. It protects the body from extrinsic factors such as ultraviolet ray, temperature change, dust, moisture, air pollution, It is a very important institution that is indispensable to.

Dermatological atrophy is a typical aging phenomenon after 70 years of age. Changes in the dermis are caused by a change in the number of fibroblasts and a change in the molecular weight of the extracellular matrix due to a decrease in their ability to synthesize. Specific changes include separation of collagen bundles, reduction of point polysaccharide synthesis, reduction of collagen and elastin number and diameter, degradation of collagen and elastic fibers, and expansion of blood vessels. Generally, among the various complex factors such as moisture content of the skin, collagen content, and immunity to the external environment, collagen degradation enzymes such as collagenase, collagenase and collagen, which reduce collagen production and collagen content, It is the active amount and activity of the genesis. Collagenase belongs to the group of matrix metalloproteinases (MMPs), which play an important role in substrate proteolysis. Such collagenase is known to decompose collagen 1, 2, 3, 5 by promoting its expression by ultraviolet rays, growth factors, inflammatory cytokines, and phorbol esters. Particularly, It is known that it plays a major role in photoaging due to the presence of light. In addition, metalloproteinases (MMPs) are composed of matrix proteins such as 4-type collagen fibers (MMP-2, -3), elastic fibers (MMP-2), or fibronectin or laminin (MMP-3) Lt; RTI ID = 0.0 > 1, < / RTI > and significantly increased expression in aged skin. Therefore, development of a material capable of reducing the activity of metalloproteinases has been actively carried out.

In addition, elastin fibers form crosslinks with collagen, and are skin constituents that are important for wrinkle formation that is involved in skin elasticity. The deficiency and aggregation of elastin fibers and the dramatic increase in activity of the elastase, elastase, have been found to be one of the causes of skin wrinkles. Elastase is the only enzyme capable of degrading elastin, and inhibition of it is known to be able to radically reduce wrinkles.

In addition, stretch marks are known in medical terms as Striae Distensae, and the precise mechanisms for their development have not yet been elucidated. In general, the most common mechanism of stretch formation is that the skin becomes excessively stretched and the collagen fibers are loosened or broken in the injured area and the skin is cleaved to form an atrophic linear band.

The stretch formed by the above-mentioned mechanism is classified into two stages: an initial stage in which atrophic pink or violet purple stripe rubra appears on the skin, and then a wrinkled and atrophied skin (Striae albae), which is the first to appear.

In addition, blue ancestors (Striae caeruleae, blue) and black ancestors (Striae nigrae, black) with increased melanization and darker color of the stretch marks are also found in later stages. The cause of stretch marks is sudden growth of body, weight gain, pregnancy and increase of body adrenocortical hormone in the adolescent period. Among them, swelling ancestor usually occurs after 6 months of pregnancy, Muscles and fats increase rapidly in the epidermis of the belly or chest. In addition, the dilated ancestors appearing in adolescents in the growing stage mainly occur around the arms, legs and groin, as the skin can not keep up with the growth rate of the musculoskeletal due to the rapid body growth rate of puberty.

As described above, dermatological treatment is most commonly used for treatment of stretch marks for various reasons. However, dermatological treatment is mainly applied to treatment of stretch marks at an early stage and does not show any effect at the late stage .

On the other hand, inflammation refers to various inflammatory mediators such as external stimuli such as tissue damage or external infectious agents (bacteria, fungi, viruses, etc.) through various pathways or internal stimuli such as in vivo metabolism, In addition to symptoms such as erythema, edema, fever, and pain associated with immune cells, it also causes various diseases such as allergies, atopy, brain diseases, circulatory disorders, and cancer. The causes of inflammation involve various biochemical phenomena. In particular, iNOS (Induced NOS) induced by stimulation of macrophages with various cytokines, TNF-alpha (Tumor necrosis factor) and LPS (Lipopolysaccharide) Nitric Oxide (NO) and Prostaglandin E2 (PGE2) are produced by Cyclooxygenase, an enzyme involved in the synthesis of prostaglandins from arachidonic acid. In addition, NO produced by NOS (NO synthase) from L-arginine is a biologically active molecule with high reactivity and functions as neurotransmitter, blood coagulation, and immune function. However, when it is produced more than necessary, it exhibits harmful effects on the living body such as excessive vasodilation, cytotoxicity and tissue damage, and wound healing inhibition. Almost all diseases that appear on the skin are accompanied by inflammation, so efforts are being made to develop a material exhibiting a anti-inflammatory effect that can be improved or treated.

1999; 江蘚新醫學院, 1979). 보통 가죽나무와 참죽나무 명칭이 혼동되어 사용되는 경우가 많은데, 이른 봄에 잎의 어린순을 부각이나 산채나물 등 요리의 재료로 이용하며 가죽나무라 일컫는 식물은 멀구슬나무과(Meliaceae)의 참죽나무(Cedrela sinensis A.)이며(Kim, 1996; 최영전, 1992), 본 연구소재인 가죽나무와는 구별되는 식물이다. 저근백피의 한방학적 효능으로는 맛은 쓰고 떫으며 성질은 서늘하고 독은 없어 열을 내리고 습을 제거하며, 설사와 출혈을 멈추게 하여 이질, 혈변 등의 지사, 지혈제로 사용하며 산후출혈, 장출혈, 위궤양 등의 치료에 사용한다(구본홍, 1994;

1999; 江蘚新醫學院, 1979). On the other hand, the leather tree ( Ailanthus altissima ) is a deciduous tree of the Simaroubaceae. It is also called a weighted tree. It is wild in all parts of the country, resistant to air pollution and insect pests, and recently it has been planted for wood, roadside trees and ornamental plants (Kim, 1996). In Oriental medicine, the root bark of the leather tree is called the hypoalbuminuric bark, the stem of the stem is called the bark, the leaf of the leather tree is the low leaf, and the fruit is the bamboo fang Grass). The herbaceous bark, which is a herbal medicine herbal medicine, is the root shell of the skin. It is used as a flavor, and its properties as a medicinal substance are cool, And hemorrhage, which are used as hemorrhagic agents such as dysentery and blood, and are used for the treatment of postpartum hemorrhage, intestinal hemorrhage, and gastric ulcer (Guobonghong, 1994;

1999; Jiang Xin New Medical Academy, 1979). In the early spring, young leaves of the leaves are used as ingredients for dishes such as reliefs and wild herbs. Plants, which are called "leather trees", are used for the cedar trees of Meliaceae ( Cedrela Sinensis A.) (Kim, 1996; Choi, Young, 1992) is a plant distinguished from the leather tree of this institute. It is used as a hemostatic agent such as dyspepsia and blood stain, and as a hemostatic agent. It is used for postpartum hemorrhage, intestinal hemorrhage, It is used for the treatment of gastric ulcer (Goo Hong, 1994;

1999; Jiang Xin New Medical Academy, 1979).

The root bark of leather wood that geunbaek blood astringent, antibacterial, antiviral, anti-inflammatory, and reported that the insecticidal action (Kim et al, 1994;. Kubota et al, 1997;.. Jeong et al, 2003), Phenolic compounds such as 3,4,5-trimethoxyphenol, p-coumaric acid, vanillin, and vanillic acid (Kazuya et al., 1994) , merosin, tannin phlobaphen, and ailanthone (Barakat, 1998). (Kim et al ., 1994), and anti-cancer activity against acute lymphoblastic leukemia (Kim and Lee et al ., 2005) , 1997;.. Hwang et al , 2001), cycle control cells (Hwang et al, 2002) and antioxidant activity (Lee et al, 2007;. Lee, 2007) represents, Chang et al. (2003) have ocotillone ingredients 70.76 % Virus cell fusion inhibitory activity. In addition, Ko and Chung et al. (1970) investigated the constituents and properties of leather seed oil, which is the origin of low-density lipoproteins for use as a fragrance resource. Lee (2008) reported on the analysis of components such as amino acids as food materials There is a bar.

Therefore, the inventors of the present invention have continued to develop a raw material for cosmetics having a skin-improving effect on a naturally occurring compound having high safety and no skin side effects. As a result, the present inventors have found that the extract of collagenase and Ela The present invention has been accomplished by discovering that it has anti-oxidizing activity to improve skin wrinkles, as well as to improve skin color and skin texture at the stretch marks area and exhibit anti-inflammatory activity.

Accordingly, a technical problem to be solved by the present invention is to provide a material having excellent human stability and having skin wrinkle or stretch marks improvement and anti-inflammatory activity.

In order to solve the above-mentioned technical problems, the present invention provides a composition for improving skin wrinkles, prevention and improvement of stretch marks, or a composition for anti-inflammation, which comprises a low molecular weight extract as an active ingredient.

Preferably, the composition comprising the low molecular weight extract of the present invention may be a cosmetic composition, a pharmaceutical composition or a food composition.

The term "extract " of the present invention means a preparation which is obtained by squeezing a herbal medicine with an appropriate leaching solution and concentrating by evaporating the leaching solution, and is not limited thereto. The extract, the diluted solution, A dried product obtained by drying, a controlled preparation thereof, or a purified product thereof.

The low molecular weight extract of the present invention can be prepared using general extraction, isolation and purification methods known in the art. As the above extraction method, there may be used a method such as hot water extraction, hot water extraction, cold extraction, reflux extraction, high temperature high pressure extraction or ultrasonic extraction. According to one embodiment of the present invention, , Reflux organic solvent extract, hot water extract or high temperature high pressure extract. These extraction methods are not particularly limited as general extraction methods known in the art.

The organic solvent may be selected from the group consisting of methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethylsulfoxide , 3-butylene glycol, propylene glycol, or a mixed solvent thereof. The extract may be extracted at room temperature or with heating under conditions where the active ingredient of the extract is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and the degree of loss of the active ingredient of the extract may differ. Therefore, it is preferable to select a suitable organic solvent.

The composition of the present invention may contain 0.1 to 90% by weight, more preferably 10 to 80% by weight, based on the total weight of the low molecular weight extract. When the content of the low molecular weight extract is less than 0.1% by weight, the skin wrinkle improving effect and anti-inflammatory effect of the present invention can not be obtained. When the content is more than 90% by weight, the effect is not proportional to the increase of the content, There is a problem that the stability of the shape is not ensured.

The composition containing the low molecular weight extract of the present invention exhibits skin wrinkle improvement, prevention and improvement of stretch marks, or anti-inflammatory effect and has little cytotoxicity as a natural substance.

According to one embodiment of the present invention, there is provided a cosmetic composition for prevention or improvement of skin wrinkles or stretch marks comprising a low molecular weight extract as an active ingredient, or a composition for anti-inflammation.

The cosmetic composition of the present invention may contain, in addition to the low molecular weight extract as an active ingredient, conventional additives such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, can do.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be prepared as a nutritional cream, a convergent lotion, a soft lotion, a lotion, an essence, a nutritional gel or a massage cream.

When the formulation of the present invention is a paste, cream or gel, use is made of an animal oil, vegetable oil, wax, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide .

When the formulation of the present invention is a powder or a spray, tosse, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Castellulose, aluminum metahydroxide, bentonite, agar or tracert, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

The cosmetic composition of the present invention may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, Any of those commonly used in ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics Such as other ingredients of the cosmetic or dermatological science. And, the above ingredients can be introduced in amounts commonly used in the field of dermatology.

According to one embodiment of the present invention, there is provided a pharmaceutical composition for skin wrinkle improvement, prevention and improvement of stretch marks, or anti-inflammation comprising an extract of low molecular weight bark as an active ingredient.

The pharmaceutical composition according to the present invention includes a pharmaceutically acceptable carrier in addition to the low molecular weight extract. The pharmaceutically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one usually used at the time of formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably applied by parenteral administration, more preferably topical application by application.

A suitable dosage of the pharmaceutical composition of the present invention may vary depending on such factors as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate, . The dosage of the pharmaceutical composition of the present invention is in the range of 0.001-100 mg / kg on an adult basis. When the composition is an external preparation, it is preferably applied in an amount of 1.0 to 3.0 ml on an adult basis once to five times a day, and continued for 1 month or more. However, the dose is not intended to limit the scope of the present invention.

The pharmaceutical composition of the present invention may be prepared in unit dosage form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art, Into the container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in an oil or aqueous medium, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

According to one embodiment of the present invention, there is provided a food composition for prevention and improvement of skin wrinkles or stretch marks, or a composition for anti-inflammation, which comprises low-coccinellar extract as an active ingredient.

The food composition according to the present invention may further comprise, as an active ingredient, not only low-molecular-weight extracts but also components normally added in the manufacture of foods such as proteins, carbohydrates, fats, nutrients, flavors and flavors .

Examples of such carbohydrates are monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings.

For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, etc., .

The composition comprising the low molecular weight extract of the present invention as an active ingredient is harmless to the human body because it contains a natural substance as an active ingredient and can be safely used for long-term use because there is little toxicity and side effects. It can be safely applied to cosmetics, pharmaceuticals and food compositions having anti-inflammatory activity as well as improvement.

As described above, the composition comprising the low molecular weight extract of the present invention as an active ingredient is excellent in anti-inflammatory activity as well as preventing and improving skin wrinkles or stretch marks, and has no cytotoxicity and skin side effects. Further, the low molecular weight extract of the present invention has no cytotoxicity and skin side effects and can be safely used in cosmetic, pharmaceutical and food compositions.

Figure 1 shows the contents of flavonoids, polyphenols and reducing sugars of the low molecular weight extract.
2 is a graph showing collagenase inhibition rates of the low molecular weight bark extract.
Fig. 3 is a graph showing the effect on the synthesis of procollagen of low molecular weight bark extract.
Fig. 4 is a graph showing the effect of the low molecular weight extract on collagen synthesis.
Fig. 5 is a graph showing the inhibition rate of elastase of the low molecular weight bark extract.
6 is a graph showing the ABTs radical scavenging activity of the low molecular weight bark extract.
7 is a graph showing the tyrosinase inhibitory activity of the low molecular weight bark extract.
8 is a graph showing the convergent activity of the low molecular weight bark extract.
9 is a graph showing the cytotoxicity of the low molecular weight bark extract.
10 is a graph showing the NO production activity of the low molecular weight bark extract.
FIG. 11 is a graph showing the amount of tumor necrosis factor-.alpha. Produced in the low molecular weight bark extract.
12 is a graph showing the amount of interleukin (IL) -1? Production of the low molecular weight bark extract.
Fig. 13 is a graph showing the amount of IL-8 produced in the low molecular weight bark extract.
FIG. 14 shows the results of ferulic acid content analysis of the low molecular weight extract from the crude extract.
15 is a graph showing the effect of the cosmetic composition containing the low molecular weight extract on the skin color of the stretch marks region.
FIG. 16 is a graph showing the effect of the cosmetic composition containing the low molecular weight extract on the skin texture of the stretch marks.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

<Example 1> Preparation of low molecular weight extract

A. altissima was produced in Korea in 2016 on herbal medicinal materials in Daegu Metropolitan City in 2017 and purchased as a herbal medicine material and finely cut and used as a sample for the extraction of low molecular weight blood.

In the case of low molecular weight bovine blood, 10 times per sample weight was added to a round flask equipped with a reflux condenser, and extraction and extraction at 80 ° C were repeated three times, and this was converted into low molecular weight reflux water (WE) .

Low molecular weight ethanolic ethanol extract (EE 70% ethanol was extracted with the solvent at 60 ℃ in the same manner as above.

Extract of water (WNPE), which is 30 times the sample weight, was extracted with solvent for 6 hours at 95 ℃.

The WHPE was extracted with water for 30 hours under the conditions of 110 ℃ and 1.5 atm.

The obtained extracts were filtered through a filter paper (Advantec No 2, Toyo Roshi Kaisha Ltd. Japan) and concentrated under reduced pressure to 1/50 using a rotary vacuum evaporator (Eyela 400 series, Tokyo, Japan) Dried powder (FD 5510 SPT, Ilsin, Korea).

&Lt; Test Example 1 > Measurement of useful ingredient content of low molecular weight bark extract

1) Content of flavonoid compound

The total amount of flavonoid compounds was determined by varying the method of Nieva Moreno et al. (2000), adding 0.4 mL of 80% ethanol, 0.1 mL of 10% aluminum nitrate and 1 mL of 1 M acetic acid to 0.1 mL of the four low- 0.1 mL of potassium and 4.3 mL of 80% ethanol were added, reacted at 25 DEG C for 40 minutes, and the absorbance was measured at 415 nm. Total flavonoid compounds were quantified from the calibration curves of quercetin (Sigma-Aldrich Co., St. Louis, Mo., USA) as the standard material.

2) Polyphenol compound content

Total polyphenol compounds contained in low molecular weight extracts with different extraction temperatures were prepared by mixing 1.8 mL of distilled water and 0.2 mL of Folin-ciocalteu's phenol reagent in 0.2 mL of the sample extracted with the Folin-Denis method (AOAC, 2005) Lt; / RTI &gt; To this solution was added 0.4 mL of 2 M Na ₂ CO _3, mixed with 1.4 mL of distilled water, and reacted for 1 hour at room temperature. The absorbance was measured at 725 nm using a spectrophotometer (Mecasys Optizen POP, Korea) The content of polyphenol compounds in the low molecular weight extract was determined by comparing with a calibration curve using tannic acid (Sigma-Aldrich Co., St Louis, MO, USA).

3) Reducing sugar content

Reduced sugar was prepared by adding 1 mL of the mixed reagent to 1 mL of the extract by Somogyi-Nelson (Nelson, 1944) method, heating it for 20 minutes, adding 1 mL of C solution, reacting at room temperature, mixing 5 mL of distilled water, Absorbance was measured. Reduced sugar content was calculated using glucose (Sigma-Aldrich Co., St. Louis, Mo., USA) to calculate the reducing sugar content of the four extracts of low-molecular-weight red blood cells.

The results are shown in Fig. Figure 1 shows the contents of flavonoids, polyphenols and reducing sugars of the low molecular weight extract. As shown here, the flavonoid contents of four extracts of low molecular weight extracts obtained under different conditions were measured. The water extract (WE) extracted at 80 ℃ showed 7.24 mg / g flavonoid and 148.11 mg / g polyphenol (EE) contained 8.59 mg / g and 165.98 mg / g, respectively. (WNPE) containing 4.8 mg / g of flavonoid and 136.89 mg / g of polyphenol were extracted under the condition of 95 ℃ for 6 hours. The low molecular weight hot press water extract (WHPE) extracted at 110 ℃ EE extracts containing 5.26 mg / g and 147.31 mg / g, respectively, contained the highest amounts of flavonoids and polyphenol compounds.

On the other hand, WNPE extracted with water at 95 ℃ for 6 hours contained the highest amount of reducing sugar. The reducing sugar was 53.03 mg / g of WE, 40.30 mg / g of EE, 171.40 mg / g of WNPE and 69.94 mg / g of WHPE.

&Lt; Test Example 2 > Measurement of skin wrinkle improving activity of the low molecular weight bark extract

1) Collagenase inhibitory activity

The collagenase inhibitory effect associated with the wrinkle formation of the skin was modified by W ㆌ nsch & Heindrich (1963) and 4 mM CaCl _{2 was added} to 0.1 M Tris-HCl buffer (pH 7.5) with 4-phenyl azobenzyloxy 0.15 mL of collagenase (0.2 mg / mL) was added to a mixture of 0.25 mL of the substrate solution in which carbonyl-Pro-LeuGly-Pro-D-Arg (0.3 mg / mL) was dissolved and 0.1 mL of the diluted sample solution And then reacted at room temperature for 20 minutes. 0.5 ml of 6% citric acid was added to the reaction mixture, and the reaction was stopped. 1.5 ml of ethyl acetate was added to the supernatant. Absorbance was measured at 320 nm using a UV / VIS spectrophotometer (Mecasys Optizen POP, Daejeon, Korea) , And the inhibition rate was expressed as the collagenase inhibition rate with the percent decrease (%) of the absorbance of the addition of the low molecular weight extract and the non-additive.

Collagen, which accounts for 70-80% of the extracellular matrix of skin dermis, is a representative connective tissue in the body and induces resistance and binding force of skin and connective tissue, mechanical rigidity, cell adhesion and support and cell differentiation Jeroma et al., 1998). Collagenase is a matrix metalloproteinase (MMPs) that specifically acts on collagen, and the expression of MMPs is increased by active oxygen and ultraviolet rays. When the expression is excessively high, collagen is excessively decomposed and deep wrinkles (Drmina & Lysenko, 1996; Fisher et al., 1999), which causes skin sagging and aging. Therefore, inhibiting the activity of collagenase may have a positive effect on delayed skin aging, especially prevention and improvement of skin wrinkles.

2 is a graph showing collagenase inhibition rates of the low molecular weight bark extract. As shown here, the collagenase inhibitory activity, which can be used as a wrinkle improving agent, was measured at a concentration of 0.1 to 1.0 mg / . The inhibition rate of WE, EE and WHPE was 46% and there was no significant difference among the four extracts. At the concentration of 0.1 mg / mL, EE extracted with 70% ethanol as solvent showed the best collagenase inhibitory activity as 27.48%.

2) Measurement of prolactin type I and collagen type I biosynthesis

When HS68 cells were treated by WNPE concentration, the amount of pro-collagen I and collagen I synthesized was measured, and the cells were planted in a 96-well plate at 1 × 10 ⁴ cells / well per well and stabilized for 24 hours. Then, the cultured medium was removed, and the WNPE extract to be used as a cosmetic raw material was treated by concentration and cultured for 48 hours. The supernatant was collected from each well, and the results were analyzed according to the procollagen type I EIA kit (FineTest) and the collagen type I EIA kit (FineTest) according to the protocol of each kit, and the absorbance was measured at 450 nm with a microplate reader .

Collagen, a major constituent of the extracellular matrix, is a major substrate protein produced by the fibroblasts of the skin. It has functions such as mechanical rigidity of the skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, induction of cell division and differentiation have. Collagen (types I, II, III, IV, and V) are synthesized in the form of precursors called procollagen. It is known that procollagen has a peptide base sequence called a propeptide at its amino terminal and carboxy terminus and has a function of cleaving and separating from collagen molecules when collagen polymerization reaction occurs. Therefore, by measuring the amount of pro peptide, the degree of collagen biosynthesis in the cell can be grasped (Parfitt et al., 1987; Kim et al., 2008b). Collagen is closely related to the formation of wrinkles in the skin and wrinkles can be caused by lack of collagen.

Fig. 3 is a graph showing the effect on the synthesis of procollagen of low molecular weight bark extract.

Fig. 4 is a graph showing the effect of the low molecular weight extract on collagen synthesis.

As shown in FIG. 3 and FIG. 4, UVB irradiation of the fibroblasts showed 28% inhibition of the prolactin biosynthesis rate compared to the normal group, and WNPE was 79, 101 and 117% at 125, 250 and 500 μg / And the collagen biosynthesis rate was 49, 72, and 85% at the normal level, indicating collagen biosynthesis. Thus, it was confirmed that WNPE significantly promotes the prolactin type I and collagen I type biosynthesis rates in HS68, one of the skin cells, in a concentration-dependent manner.

3) Elastase inhibitory activity

Elastase inhibitory activity of the four extracts of low molecular weight peptides was determined by Cannell et al. 0.5 mL of porcine pancrease elastase (2.5 U / mL) dissolved in 50 mM Tris-HCl buffer (pH 8.6) and 50 mL of 50 mM (L-Ala) ₃ -p-nitroanilide (0.5 mg / mL) was added to Tris-HCl buffer solution at 25 ° C for 20 minutes to prepare p -Nitroanilide was measured at 405 nm. The inhibition rate of the absorbance of the low molecular weight extract and the low density lipoprotein of the low molecular weight extract was expressed as percentage (%) and expressed as the inhibition rate of elastase.

Elastase is a nonspecific hydrolytic enzyme capable of degrading various proteins including elastin, pibro-pectin and collagen, which are matrix proteins that maintain skin elasticity in the dermis, and is a direct cause of wrinkle formation due to skin cell destruction (Balo & Banga, 1950; Tsukahara et al., 2006). Since the elastase inhibitor improves the wrinkles of the skin and plays an important role in suppressing the aging, uric acid is widely used as an elastase inhibitor. However, it is difficult to formulate uric acid because it has insoluble properties in solvents such as water and oil (Lee & Lee, 2008).

Fig. 5 is a graph showing the inhibition rate of elastase of the low molecular weight bark extract. As can be seen from the results, the inhibition rate of the elastase for wrinkle improvement of the four extracts of low molecular weight bark was found to be 36.15% at a concentration of 1.0 mg / mL, Inhibition rate of EA, WNPE 18.02%, and WHPE 15.69% of EE extracted with 70% ethanol as a solvent.

&Lt; Test Example 3 > Measurement of antioxidative activity of low molecular weight extract

The elimination ability of ABTS [2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)] radicals was measured by modifying the method of Re et al. (1999). 7 mM ABTS and 2.45 mM potassium sulfide were added and reacted at room temperature for 12-16 hours to produce ABTS cations. The absorbance at 734 nm was adjusted to be 0.70 ± 0.02 by using 99.9% ethanol. Then 100 μL of ABTS solution was added to 100 μL of the sample, reacted at room temperature for 7 minutes, and the absorbance was measured at 734 nm. The ABTS radical scavenging activity (%) was calculated from the absorbance ratio of the sample to which no sample was added.

The ABTs radical scavenging activity is determined by the fact that the cationic ABTS radicals produced by the reaction with potassium persulfate are removed by the antioxidative substances contained in the extract and the radical specific cyan color is discolored to light green due to the radical inhibition action of the antioxidant Antioxidant activity can be measured without distinguishing between polar and nonpolar samples (Awika et al., 2003).

6 is a graph showing the ABTs radical scavenging activity of the low molecular weight bark extract. As shown here, the ABTs radical scavenging activity of the four extracts of low molecular weight bacteriocin was 52.66% ~ 59.68% at 100 μg / mL, which was lower than that of the control L-ascorbic acid, WHPE extracts at high temperature and high pressure showed the best ABTs radical scavenging ability.

Test Example 4 Whitening activity of low molecular weight bark extract

The tyrosinase inhibitory activity, which is one of the methods to confirm the whitening activity of the low molecular weight extract, was measured according to the method of Yagi et al. (1986). A substrate solution in which 10 mM of 3,4-dihydroxy-L-phenylalanine (L-DOPA; Sigma-Aldrich Co., St Louis, MO, USA) was dissolved in 0.5 mL of a 175 mM sodium phosphate buffer solution (pH 6.8) 0.2 mL of mushroom-type tyrosinase (110 U / mL) was added to the mixture of 0.2 mL of the sample and 0.1 mL of the diluted sample at a constant concentration. After reacting at 25 ° C for 2 minutes, the resulting DOPA chromium was absorbed at 475 nm , And the inhibition rate of tyrosinase was expressed as a percentage (%) of the absorbance of the sample without and with no addition.

Tyrosinase contains Cu ²⁺ , which uses a wide range of phenolic compounds as a substrate, and is a major enzyme that produces dark brown melanin, a pigment component of the skin. When tyrosinase activity is inhibited, melanin production (Imokawa & Mishima, 1980; Laskin and Piccinini, 1986). Melanin is a polymeric substance of phenols widely distributed in nature, and biosynthesis is promoted by a mechanism against ultraviolet rays in human skin (Yoneta et al., 2004). The melanin pigment blocks ultraviolet absorption and protects the skin by increasing the resistance. However, the melanin pigment itself generates active oxygen. Excessive melanin pigment formation is caused by pigmentation such as spots, freckles, skin spots and toxicity of melanin precursors (Kameyama et al., 1993). In addition, it is possible that skin cancer and skin aging may be promoted.

7 is a graph showing the tyrosinase inhibitory activity of the low molecular weight bark extract. As shown in the table, tyrosinase inhibitory activity of tyrosinase inhibiting tyrosinase inducing melanin pigmentation was evaluated in a concentration of 1.0 mg / mL of WNPE 53.20%> WHPE 49.09%> WE> 47.74%> EE 43.06% of water extracts showed slightly higher tyrosinase inhibitory activity than ethanol extracts.

&Lt; Test Example 5 > Measurement of astringent activity of low molecular weight blood extract

Astringent activity was measured according to the method of Lee et al. (2002). Hemoglobin similar to the skin protein was used as the reaction mixture. Each sample solution and hemoglobin solution (Sigma-aldrich Co.) were added to the centrifuge vessel in a ratio of 1: 1, , And then the absorbance was measured at 576 nm after centrifugation at 5000 rpm for 5 minutes. Convergence activity was calculated according to the following equation (1).

Convergence basically means wrinkle or shrink, and the principle of convergence is the phenomenon that the skin contracts due to the skin protein forming the crosslinking with the polymer flavonoid, and the astringent agent has excellent protein binding properties. Such convergent action has the effect of forming an insoluble film on the surface of skin and mucous membrane by external use or contents and consequently protecting the locality or densifying the tissue to reduce the permeability of the cell membrane. Skin and pores that have lost elasticity are given a convergence effect on the enlarged skin, giving the skin a sense of tension and elasticity at the same time. Using hemoglobin, a pigment protein, the degree of binding of hemoglobin to the extract was measured to determine the convergence effect.

8 is a graph showing the convergent activity of the low molecular weight bark extract. As shown in the graph, the convergence effect of EE was the highest at 44.24% at the concentration of 100 μg / mL, and that of WNPE and WHNPE was 36.34% and 31.93% Effect.

&Lt; Test Example 6 > Cytotoxicity measurement of low molecular weight blood extract

The mouse macrophage cell line RAW264.7 and the human keratinocyte cell line HaCaT cells were treated with 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic (10 units / mL penicillin, 100 μg / mL streptomycin, 0.25 μg / (Forma series II, Water Jacketed M311, Thermo Electron Co., USA) fed with 5% CO ₂ at 37 ° C in Dulbecco's modified Eagle's medium (DMEM) containing Dulbecco's modified Eagle's medium The cytotoxicity and nitric oxide (NO) production inhibitory effect of low molecular weight extracts were investigated.

The cytotoxicity of the low molecular weight extracts from RAW264.7 macrophages and HaCaT keratinocytes using MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) Were measured. HaCaT and RAW264.7 cells were 1X10 ⁵ (Falcon Co., USA) at a concentration of 10 &lt; RTI ID = 0.0 &gt; ml / ml &lt; / RTI &gt; After 24 hours, the cells were treated with 100 μL of DMEM medium containing low-molecular weight extracts diluted to a constant concentration for 24 hours. Then, 5 μg / mL of 3- (4,5-dimethyl (100 μL each) was added to each well of a 5% CO ₂ incubator at 37 ° C. in a 3% (v / v) Lt; / RTI &gt; After removing after the culture supernatant and dissolved formazan was added to 100 μL DMSO (dimethyl sulfoxide) to each well ELISA (Xmark microplate absorbance spectrophotometer ^(Bio-Rad, XMark ^TM, absorbance at 570 nm using a Japan) And the cell viability for the control group was expressed as a percentage.

9 is a graph showing the cytotoxicity of the low molecular weight bark extract.

MTT assay was used to confirm the cytotoxicity of four low molecular weight extracts from RAW264.7 macrophages and dermal keratinocytes, HaCaT cells. The cell viability of Raw 264.7 cells was about 55% at 100 μg / mL of ethanol extract, EE, and that of WE, WNPE (100 μg / mL) And WHPE extracts showed cell viability above 85%. In HaCaT cells, cell viability of 85% or more was observed at 100 μg / mL, and the low-molecular-weight extract of water-extracted cells showed a safe concentration of 100 μg / mL, Respectively.

< Test Example  7> Low-grade blood  Determination of anti-inflammatory activity of extract

1) NO production measurement

RAW 264.7 macrophages were plated on 24-well cell culture plates at 2.5 × 10 ⁵ cells / mL per well and incubated for 18 hours at 37 ° C. in a 5% CO ₂ incubator. The cultured cells were exchanged with medium containing 1 μg / mL of lipopolysaccharide (LPS) to induce the production of NO, and then cultured for 24 hours with addition of low molecular weight extract. Subsequently, the amount of NO produced was determined by mixing 100 μL of the cell culture supernatant with 100 μL of Griess reagent (0.1% naphthylethylenediamine dihydrochloride and 1% 1% sulfanylamine in 2.5% phosphoric acid, 1: 1) Min, and the absorbance was measured at 540 nm using an ELISA reader (Xmark microplate absorbance spectrophotometer, Bio-Rad, XMark ^™, Japan). The amount of NO produced was calculated by comparing with the standard curve for the concentration of sodium nitrate (NaNO ₂ ).

NO is one of the reactive oxygen-enriched active oxygen species, which is synthesized by NOS as L-citrulline with L-arginine as a substrate. It can kill bacteria or induce apoptosis, signal transduction, blood coagulation and platelet suppression , Neurotransmitters involved in vasodilation, and so on. However, over-formed NO can cause inflammation leading to tissue damage, genetic mutations, nerve damage, and immune disorders such as arthritis and bronchitis (Weise & Nathan 1991; Hyun et al., 2015).

10 is a graph showing the NO production activity of the low molecular weight bark extract. As shown here, to investigate the effect of low molecular weight extracts on the production of NO, which is known to play an important role in inflammation induction in the four extracts of low molecular weight bacterium, the nitroxide (nitro oxide) NO production in one mouse macrophage RAW264.7 cell was determined. As a result, the LPS - treated group showed a higher NO expression level than the LPS - treated group, and the NO expression was decreased in the group treated with the low molecular weight extract. WNPE extracted with water at 95 ℃ and WHPE extracted at 110 ℃ with 1.5 atmospheric pressure showed inhibition rates of 14% and 12%, respectively.

2) Measurement of cytokine production

Enzyme-Linked Immunosorbent Assay (ELISA) To examine the effect of low-molecular weight extracts on the production of inflammatory cytokines such as TNF-α and IL-8, LPS (1 μg / (25, 50, and 100 μg / mL), and cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours. Then, the culture solution was recovered to obtain the respective TNF-α, IL- IL-8 content was measured by ELISA cytokine kit (R & D system, Minneapolis, MN, USA) according to manufacturer's instructions.

 The standard solution (100 μL) and the cell culture solution (100 μL) were added to a 96-well plate coated with the antibody, reacted at room temperature for 2 hours, washed with a washing solution, and 100 μL of a detection antibody . 100 μL of avidin-HRP was added to each well, followed by reaction for 30 minutes. After washing with a washing solution, 100 μL of TMB was added, and the reaction was allowed to proceed for 20 minutes in a dark room. The secretion of cytokines was calculated by measuring the absorbance at 450 nm.

The macrophage is involved in innate immunity and acquired immunity, and maintains the homeostasis. When the antigen enters the body, it removes the antigen by phagocyte action or induces the antigen to appear on other immune cells to induce the inflammation reaction. (Lee et al., 2007). The macrophages are activated by immune enhancing substances such as interferon (IFN) and glycolipid (LPS), and are involved in tumor necrosis factor-α (TNF-α), interleukin , Nitric oxide (NO), and the like (Hibbs et al., 1998). TNF-α is a multifunctional cytokine that is secreted when mast cells are activated and is involved in the immune and inflammatory responses and promotes the secretion of other inflammatory cytokines such as IL-1β, IL-6, and IL-8 Adcock, 1993; Butler et al., 1995). IL-8 is a chemoattractant that promotes the mobilization of neutrophils, lymphocytes, and eosinophils, activates neutrophils to release lysosomal enzymes, induce active coral production, and is known to be involved in inflammatory diseases such as arthritis and sepsis (Harada et al., 1996). In addition, IL-1β is a cytokine produced by monocytes and macrophage-derived cells, which activates T cells and promotes the activity of cytokines such as IL-2 secreted from T cells. (Cah et al., 2010).

The secretions of cytokines (TNF-α, IL-1β, IL-8) secreted from the culture supernatant of RAW 264.7 macrophages treated with LPS (1 μg / Respectively.

FIG. 11 is a graph showing the amount of tumor necrosis factor-.alpha. Produced in the low molecular weight bark extract.

12 is a graph showing the amount of interleukin (IL) -1? Production of the low molecular weight bark extract.

Fig. 13 is a graph showing the amount of IL-8 produced in the low molecular weight bark extract.

As shown in FIGS. 11 to 13, in the case of TNF-α, WE, EE and WNPE showed a inhibition rate of about 23 to 30% at a concentration of 100 μg / mL, and a WHPE inhibition rate of 19%. In the case of IL-1β, ethanol extract EE showed the highest inhibition rate at 100 μg / mL and water extract showed similar inhibitory activity within about 20%. The inhibitory activity of IL-8 on RAW 264.7 cells stimulated by LPS was inhibited by 45-66% of IL-8 in all extracts regardless of concentration.

3) Ferulic acid content analysis

UPLC analysis was performed for the analysis of ferulic acid, which is a low molecular weight component. The UPLC was a Waters H-Class and the column was C ₁₈ (4.6 x 150 mm, 2.7 μm, CORETECS ㄾ). The detector was UV-280 nm, the gradient was acetonitrile and water, and the flow rate was 0.25 mL / min. For the preparation of the standard solution, 4.0 mg of ferulic acid as a standard was accurately weighed and dissolved in 1.0 mL of acetonitrile for HPLC. The diluted solution was diluted to 15.625, 31.25 62.5, and 125 μg / mL with a stock solution and used as a standard solution Prepare the calibration curve and analyze the sample solution of leather wood from the prepared calibration curve by 5.0 μL of each UPLC condition. The peak area corresponding to each standard product is substituted into the y value of the regression equation to obtain the concentration of each compound Respectively.

FIG. 14 shows the results of ferulic acid content analysis of the low molecular weight extract from the crude extract. The linearity was confirmed as a result of diluting standard substances by concentration. The concentration of ferulic acid was estimated to be about 62.5 μg / mL in the range of 25 to 200% with the target concentration set at 100%. As a result, the linearity was confirmed by concentration and y = 39258X + 32222 (R ₂ = 0.9999) appear. The peak area of the compound was 76415, and the peak area corresponding to the compound was substituted for the y value of the regression equation to determine the concentration of ferulic acid. As a result, It was analyzed that ferulic acid was contained at 70 μg / g per 1 mg of leather wood sample.

&Lt; Test Example 8 > Safety confirmation test for human skin of low molecular weight bark extract

In order to confirm the safety of the extract from the skin, it was investigated whether or not the extract of the low molecular weight extract stimulates the skin for 48 hours in a healthy 30 adults. 20 μL of the test substance was dropped into the IQ chamber, and the test site of the study subject was wiped with 70% ethanol, followed by drying and adhering. The test material was closed for 48 hours. After removal of the patch, the test area was marked with a skin marker. After 30 minutes and 24 hours of removal of the patch, the skin reaction under a magnifying glass (SK101-3X, SeKi optical, Respectively.

Skin response evaluation was evaluated according to the criteria shown in Table 1 below, which reflected Frosch & Kligman 1979, PCPC Guidelines and ICDRG.

The skin response to each substance at 30 minutes and 24 hours after the removal of the patch was calculated according to the following formula (2), and the skin irritation was judged according to the following Table 2 based on this. The results are shown in Table 3 below.

As shown in the above Table 3, no skin reaction was observed in all subjects participating at the time of observation at 30 minutes and 24 hours after the removal of the patch.

&Lt; Test Example 9 > Measurement of improvement effect of stretch marks on low-coccinellar blood extract

In order to measure the skin color and skin texture improving effect of the stretch marks of the cosmetic composition containing the low molecular weight extract, the cream containing the low molecular weight extract was prepared by the method of Preparation Example 1-1, Twenty-three women with stretch marks were allowed to use the cream for 4 weeks. The subjects forbid the use of body products 12 hours before the visit. Twice a day (morning and evening) Apply the appropriate amount of sample to the designated application site (left or right) and absorb it sufficiently.

In the human body test, the test area was washed under the conditions of no temperature and humidity (20 ~ 24 ㅀ C, 40 ~ 60% RH) without air movement and stabilized for 30 minutes before proceeding. The test sites were designated as non-adjacent stretch sites and sample application sites, respectively. The sample application site was designated as the left and right side of the subject. Acetate film was adhered to the designated test site and the points were marked so that the same site could be measured at each measurement.

1) Evaluation of skin color in stretch zone using a chroma meter

The chroma meter CR-400 is a device that converts colors recognized by humans into digital codes composed of three factors. It is widely used for skin color measurement in cosmetics research. The a * value is a chrominance parameter. To evaluate the degree of redness of the stretch marks due to swelling, the mechanical evaluation was performed by measuring the test area three times with a chroma meter and then averaging the a * values. The reduction rate of the a * value using a chromatic meter was calculated using Equation (3).

The results of the measurement of the a * value of the stretch zone using the chromatic meter are shown in Tables 4 to 8 (Table 4: Results of end-to-end endurance test, Table 5: Table 7: Results of pre-homogeneity test of the a * value of a * values, Table 7: Pre-homology test results of a * values between test sites. : the percentage of subjects whose a * values were reduced (%)).

In addition, the weekly effect of the sample application site was statistically significant ( p <0.01 ) as a result of the endogenous effect test before and after application of the sample to the a * value of the stretch zone using the chroma meter.

As a result of the individual-contrast test, the a * value of the sample application site was statistically significant ( p <0.01 ) compared to that before application and decreased by 1.718% after 8 weeks of application.

The homogeneity test was performed to confirm the difference of a * value between test sites before application of the sample. As a result, there was no statistically significant difference in the a * value between the sample application site and the unused site.

The results of Site - Week interaction effect of a * value showed statistically significant difference ( p <0.01 ).

15 is a graph showing the effect of the cosmetic composition containing the low molecular weight extract on the skin color of the stretch marks region. As a result of the contrast test between the groups, the a * value of the sample application area showed a statistically significant decrease (p <0.01) after 8 weeks of application compared to the unused area.

As a result of analyzing the ratio of the subjects whose a * value was decreased, the a * value of the sample application site decreased by 82.609% after 8 weeks of application.

2) Evaluation of skin texture of stretch area using PRIMOS Lite

end. Measurement of skin texture of stretch marks using PRIMOS Lite

PRIMOS Lite (Phase Shift Rapid in vivo Measurement of Skin, GFMesstechnik GmbH, Germany). To measure the same area at each measurement, the equipment was measured at the same distance and height at the test site.

I. Analysis of skin texture of stretch marks using image analysis

The skin texture was analyzed using PRIMOS software (PRIMOS version 5.8E). The three-dimensional measurement of PRIMOS Lite changes the parallel projection stripes projected onto the skin according to the height difference of the skin surface, and the degree of change is quantitatively computed by computer. The image of the stretch zone measured by PRIMOS Lite was matched to the measurement site using 3D matching. The values of the skin texture parameters from the roughness analysis are as follows (Ra (average roughness): average value of the absolute values of the lengths from the center line to the surface curves of the surface when drawing the center line at the roughness height of the section; Which means that the skin texture is improved).

The reduction rate of the Ra value due to the PRIMOS high resolution was obtained using the equation (4).

All. Evaluation of the proportion of subjects whose measured values were reduced compared to those before the sample application

The ratio of the subjects whose measured values were decreased after application of the sample was obtained by using the following equation (5).

The results of the skin roughness measurement, reduction rate and statistical analysis of the stretch zone using PRIMOS Lite are shown in the following Tables 11 to 17 (Table 11: Results of end-to-end endurance test, Table 12: Table 13: Average of reduction rate of individual Ra value (%), Table 14: Average of decrease rate (%) of individual Ra value, Table 15: : Inter-group individual-internal comparison test, Table 17: Ratio of subjects with reduced Ra (%).

As a result of the pre - and post - treatment tests of the Ra value of the stretch zone using PRIMOS Lite, the Week effect of the sample application area showed a statistically significant difference ( p <0.001 ).

As a result of the individual-contrast test, the Ra value of the sample application site was statistically significant ( p <0.01 ), and decreased by 5.120% and 8.552% after 4 weeks and 8 weeks after application . FIG. 16 is a graph showing the effect of the cosmetic composition containing the low molecular weight extract on the skin texture of the stretch marks.

The homogeneity test was conducted to confirm the difference of Ra values between the test sites before application of the sample. As a result, the Ra values of the applied region and the unused region were not statistically significant.

The results of Site - Week interaction effect on the Ra value were statistically significant ( p <0.01 ).

As a result of the contrast test between groups, the Ra value at the application site showed a statistically significant decrease (p <0.01) after 8 weeks of application compared to the unused area.

As a result of the analysis of the ratio of the subjects whose Ra values were decreased, the subjects of the sample application areas showed a decrease in Ra value after 4 weeks of application and after 8 weeks of application, 69.565% and 91.304%, respectively.

3) Sensory evaluation

After 4 weeks of application and after 8 weeks of application, subjects were asked to rate their likelihood of evaluating the efficacy of the samples.

The results are shown in the following Tables 18 to 20 (Table 18: Effect of improving stretch marks, Table 19: Effect of improving skin texture of stretch marks, and Table 20: Improvement of redness of stretch marks).

As shown in Tables 18 to 20, the sensory evaluation of the cosmetic composition containing low-molecular-weight extracts showed that the effect of improving the stretch marks, improving the skin texture of the stretch marks and improving the redness of the stretch marks were excellent.

&Lt; Formulation Example 1 > Preparation of cosmetic material

1-1. Manufacture of cream

As shown in the following Table 21, a cream containing the low molecular weight extract as an active ingredient was prepared by a conventional method.

ingredient Content (% by weight) Low molecular weight extract 0.03 or 0.01 Carbomer 940 0.12 Trehalose 3.00 Sorbitol 1.00 1,3-butylene glycol 5.00 Bis-phage-18 methyl ether dimethylsilane 1.00 Glycerol 4.00 DIEL - Panthenol 0.30 Edith Aie 0.02 Polysol Bait 60 0.80 Glycerol monostearate 1.20 Glycerol stearate / phage-100 stearate 0.90 Solvant Seth Quilletate 0.30 Rannet-O 1.60 Cetyl ethylhexanoate 3.00 Caprylic / Carrick Triglyceride 1.00 Cyclomethicone 6.50 Tririthanolamine 0.13 Pernonip 0.70 Incense, preservative a very small amount Purified water Balance Sum 100

1-2. Manufacture of Toner

Toners containing low molecular weight extracts as active ingredients as shown in Table 22 below were prepared according to a conventional method.

ingredient Content (% by weight) Low molecular weight extract 0.05 or 0.01 Dysodium EDTA 0.01 Amino coat 1.00 DIEL - Panthenol 0.10 Trehalose 0.10 Glycerol 3.00 Carbopol ITD 2020 0.07 Alcohol 3.00 Bis-phage-18 methyl ether dimethylsilane 0.20 Phenyl trimethicone 0.10 Phage-40 Hydrogenated Cosol Oil 0.15 Methyl paraben 0.05 Triethanolamine 0.13 Pernonip 0.70 Phenoxyethanol 0.50 Fragrance, preservative, pigment a very small amount Purified water Balance Sum 100

1-3. Manufacture of lotions

A lotion containing the low molecular weight extract as an active ingredient was prepared according to a conventional method as shown in Table 23 below.

ingredient Content (% by weight) Low molecular weight extract 0.05 or 0.01 Carbomer 941 0.05 Allantoin 3.00 Hub X-Hyaluron 1.00 Methyl paraben 0.10 Glycerol 1.10 Dysodium EDTA 0.05 Polysorbate 60 1.00 Solitanseciolate 1.00 Stearic acid 1.00 Glycerol stearate / phage-100 stearate 1.50 Cetyl alcohol 1.30 Propylparaben 0.05 Butyl paraben 0.05 Macadamia oil 0.05 Dimethicone 0.50 Caprylic / capric triglyceride 1.00 Mineral oil 4.50 Triethanolamine 0.10 Phenoxyethanol 0.50 incense a very small amount Purified water Balance Sum 100

Claims (3)

A cosmetic composition for skin wrinkle improvement, prevention and improvement of stretch marks, or anti-inflammation comprising low molecular weight bark extract as an active ingredient. A pharmaceutical composition for improving skin wrinkles, prevention and treatment of stretch marks, or antiinflammation, which comprises low molecular weight bark extract as an active ingredient. A composition for improving skin wrinkles, prevention and improvement of stretch marks, or a composition for anti-inflammation, which comprises low molecular weight bark extract as an active ingredient.

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