KR102741204B1 - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. A mixed extract of an extract of Cheonmun-dong, an extract of Rehmannia glutinosa, and an extract of Baekbokryeong according to the present invention significantly reduces the total amount of melanin, thereby exhibiting a whitening effect, promotes collagen synthesis of skin fibroblasts, inhibits elastase activity, thereby improving wrinkles and enhancing elasticity, inhibits nitric oxide (NO) production, thereby exhibiting an anti-inflammatory effect, and through an antioxidant effect by scavenging free radicals and an antiglycation effect by inhibiting glycation, can be used in the manufacture of medicines, cosmetics, or foods for skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant, and/or antiglycation.

Description

Composition for improving skin

The present invention relates to a composition for improving skin, which exhibits skin whitening, elasticity enhancement, wrinkle improvement, anti-inflammation, antioxidant, and anti-glycation effects.

It is a common desire to have fair and fair skin. The color or brightness of the skin is genetically determined by the concentration and distribution of melanin in a person's skin, but it is also affected by environmental or physiological conditions such as ultraviolet rays from the sun, fatigue, or stress. Melanin is created by sequentially converting tyrosine, a type of amino acid, into DOPA and dopaquinone through an enzyme called tyrosinase, which acts as a catalyst, and then undergoing a non-enzymatic oxidation reaction. The pathway by which melanin is created is known, but the cause of tyrosinase being triggered in the mechanism that induces melanin synthesis is still not clearly known.

Meanwhile, commonly known whitening ingredients include substances that inhibit tyrosinase enzyme activity, such as kojic acid or arbutin, hydroquinone, vitamin C (L-ascorbic acid) or their derivatives, and various plant extracts. These can achieve skin whitening by brightening the skin tone by inhibiting the synthesis of melanin pigment, and can also improve skin hyperpigmentation such as blemishes and freckles caused by ultraviolet rays, hormones, or genetics. However, when applied to the skin, there is a problem that the amount used is limited due to safety issues such as irritation and redness, or the effect is minimal and no actual effect can be expected.

In addition, collagen is a major matrix protein produced by fibroblasts in the skin and exists in the extracellular matrix, and its important functions are known to be mechanical strength of the skin, resistance of connective tissue, cohesion of tissues, support of cell adhesion, and induction of cell division and differentiation (growth of organisms or wound healing). It is known that this collagen decreases with age and photoaging caused by ultraviolet irradiation, and this is closely related to the formation of wrinkles in the skin. In addition, as extensive research on skin aging has been developed in recent years, the important functions of collagen in the skin are being revealed.

Effective ingredients that promote collagen synthesis and have an effect of improving wrinkles are known. For example, retinoic acid, TGF (transforming growth factor) [Non-patent document 1], protein derived from animal placenta [Patent document 1], betulinic acid [Patent document 2], chlorella extract [Patent documents 3, 4], etc. are known as substances that promote collagen synthesis. However, the above effective ingredients have limitations in the amount of use due to safety issues such as irritation and redness when applied to the skin, or have minimal effects, so there is a problem that it is not possible to expect the effect of improving skin function by actually promoting collagen synthesis in the skin.

In addition, inflammation is an immune response of the human body in response to injury or disease, and oxidative stress such as ultraviolet rays, active oxygen, and free radicals activate inflammatory factors, causing various diseases and skin aging. Vasoactive polypeptides such as kinin, plasmin, and complement cause vasodilation and contraction and chemotaxis, and lymphokines such as interleukin-6 (IL-6) and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized into prostaglandins or leukotrienes, which are inflammatory mediators, through two pathways, cyclooxygenase or lipooxygenase, and mediates various inflammatory responses.

An anti-inflammatory agent is one that works to eliminate the source of inflammation, reduce biological responses, and reduce symptoms in order to eliminate inflammation. Currently, non-steroidal substances such as flufenamic acid, ibuprofen, benzydamine, or indomethacin are used for anti-inflammation purposes, and steroidal substances such as prednisolone or dexamethasone are used. In addition, allantoin, azulene, and hydrocortisone are known to be effective in anti-inflammation, but these substances have limitations in the amount of use due to safety issues on the skin, or they have minimal effects, making it difficult to expect a practical anti-inflammatory effect.

Meanwhile, active oxygen that is introduced from outside the body or generated inside the body causes many problems such as accelerating the aging of the body or causing cancer. Therefore, much development and research is being done on antioxidant substances that suppress oxidation by active oxygen. Antioxidants are widely distributed in the animal and plant kingdoms, and many phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium are known to be present in fruits and vegetables. However, it is difficult to expect sufficient effects from antioxidant substances that exist naturally when applied to the skin. Therefore, synthetic antioxidants with excellent antioxidant properties and low prices are widely used, but their use is limited due to safety concerns such as side effects on the human body.

Glycation generally refers to a reaction in which simple sugars such as glucose or fructose form covalent bonds with proteins or fats without the involvement of enzymes. The accumulation of advanced glycation end products changes proteins into a hard and more brittle state, and glycated collagen prevents collagen from forming a proper structure in the extracellular matrix of the dermis, thereby causing skin to lose elasticity and promoting wrinkle formation [Non-patent document 2]. In addition, advanced glycation end products generated by glycation are brownish substances, and are thought to be the cause of the skin complexion gradually turning yellow as skin ages, and it is known that the more advanced glycation end products accumulate, the less reflected light from the skin is [Non-patent document 3]. Aminoguanidine, pyridoxamine, and aspirin are known as substances that inhibit glycation, but these substances have limitations in the amount of use due to safety issues on the skin, or their effects are minimal, making it difficult to expect a practical effect.

In addition, there is an urgent need for the development of ingredients that are safe for the body, have stable active ingredients, and, above all, have skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, and/or antioxidant activities that are more effective than existing skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, and/or antioxidant effects.

Meanwhile, Asparagus cochinchinensis MERR is the tuberous root of Asparagus cochinchinensis Merrill (Liliaceae), which is boiled or steamed in hot water, the outer skin is removed, and then dried. Its effective ingredients include asparagine and beta-sitosterol. In animal experiments, diuretic and tonic effects were confirmed, and in vitro, it showed an antibacterial effect against Staphylococcus aureus, hemolytic Streptococcus, Diplococcus pneumoniae, and Diphtheria spp.

Also, Rehmannia Radix Preparat is the root of Rehmannia glutinosa Liboschitz ex Steudel (Scropophulariaceae). Rehmannia Radix Preparat is the main medicinal ingredient of Samultang (四物湯) and is used for symptoms such as internal fever, dry throat, and thirst that occur due to weakness in various chronic diseases.

Also, poria sclerotium is the sclerotium of Poria cocos Wolf (Polyporaceae family).

Japanese Special Publication No. 8-231370 Japanese Special Publication No. 8-208424 Japanese Special Publication No. 9-40523 Japanese Special Publication No. 10-36283

Cardinale G. et al, Adv. Enzymol., 41, p. 425, 1974 Dyer, D. G. et al, The Journal of Clinical Investigation., 9, p. 2463, 1993 Hiroshi, O. et al, Skin Research and Technology, 15, p. 496, 2009

Accordingly, the inventors of the present invention have completed the present invention by confirming that a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract significantly reduces the total amount of melanin, exhibits a whitening effect, promotes collagen synthesis of skin fibroblasts, inhibits elastase activity, improves wrinkles and increases elasticity, exhibits an anti-inflammatory effect by inhibiting nitric oxide (NO) production, and exhibits an antioxidant effect that scavenges free radicals and an antiglycation effect that inhibits glycation.

Accordingly, the purpose of the present invention is to provide a composition for skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, anti-oxidation and/or anti-glycation, which contains as an effective ingredient a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract and Baekbokryeong extract.

As a means for solving the above problems, the present invention provides a composition for skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, anti-oxidation and/or anti-glycation, comprising as an effective ingredient a mixed extract of an extract of Cheonmun-dong, an extract of Rehmannia glutinosa and an extract of Baekbokryeong, for the manufacture of a medicine, cosmetic or food.

The mixed extract of the extract of Cheonmun-dong, the extract of Rehmannia glutinosa and the extract of Baekbokryeong according to the present invention significantly reduces the total amount of melanin, thereby exhibiting a whitening effect, promotes collagen synthesis of skin fibroblasts, inhibits elastase activity, thereby improving wrinkles and enhancing elasticity, exhibits an anti-inflammatory effect by inhibiting nitric oxide (NO) production, and exhibits an antioxidant effect and an antiglycation effect by scavenging free radicals, thereby being usable in the manufacture of pharmaceuticals, cosmetics or foods.

Hereinafter, the composition of the present invention will be described in detail.

In order for a skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation ingredient to exhibit excellent effects when applied to actual skin, it is desirable that the ingredient exhibit highly active skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation activities at a low concentration, have excellent ability to penetrate and be absorbed through the skin, have low volatility so that they can remain for a sufficient time to exhibit skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation effects, be stably maintained as an active ingredient on the composition or skin, be easy to manufacture into a pharmaceutical, cosmetic or food, and be safe for the skin. However, it is rare for a known ingredient to satisfy all of the above characteristics. For example, some skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation ingredients exhibit excellent skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation activities even at low concentrations in in vitro experiments, but have poor ability to penetrate and be absorbed through the skin, making it difficult to apply to actual skin. Other active ingredients have low hydrophilicity, making them difficult to formulate into medicines, cosmetics, or foods. In addition, some skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, antioxidant, and/or anti-glycation ingredients may decompose or transform into other compounds when exposed to heat, light, or oxygen, thereby losing their effectiveness before they are applied to the skin.

As can be seen in the examples below, the extracts of Cheonmun-dong, Rehmannia glutinosa and Baekbokryeong can be used as effective ingredients of pharmaceutical, cosmetic or food compositions for the excellent effects of reducing the total amount of melanin, anti-glycation, promoting collagen synthesis, anti-inflammation, anti-oxidation and/or anti-glycation at low concentrations.

Accordingly, the present invention provides a composition for skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, anti-oxidation and/or anti-glycation, which contains as an effective ingredient a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract and Baekbokryeong extract.

In the present invention, Asparagus cochinchinensis MERR is a perennial herbaceous plant belonging to the lily family. In addition, Rehmannia glutinosa is a herbal medicine obtained by steaming and drying the root of Rehmannia glutinosa (Gaertner) Libosch. Baekbokryeong is the dried tuber of Poria cocas Wolf.

The above extracts of Cheonmun-dong, Sukjihwang, or Baekbokryeong can be extracted by methods known in the art, and the methods are not particularly limited. Alternatively, commercially available extracts can be used.

Preferably, the extract of Cheonmun-dong, Rehmannia glutinosa or Poria cocos may be an extract obtained by extracting Cheonmun-dong, Rehmannia glutinosa or Poria cocos with water and/or an organic solvent, and the organic solvent may be a polar organic solvent, a non-polar organic solvent or a mixed solvent thereof. The polar organic solvent may be a lower alcohol having 1 to 5 carbon atoms, ethyl acetate or acetone, and the non-polar organic solvent may be ether, chloroform, benzene, hexane or dichloromethane. For example, the lower alcohol having 1 to 5 carbon atoms may be methanol, ethanol, propanol, butanol or isopropanol.

In one specific example, the Cheonmun-dong extract, the Rehmannia glutinosa extract, or the Poria cocos extract may include a fraction obtained by fractionating a primary extract extracted using the aforementioned extraction solvent using an extraction solvent having a different polarity. For example, the Cheonmun-dong extract, the Rehmannia glutinosa extract, or the Poria cocos extract may be a fraction extracted with an alcohol having 1 to 5 carbon atoms, and then fractionated again with a solvent having a different polarity, such as ether, benzene, or hexane. Two or more solvents may be used in the fractionation, and each solvent extract may be prepared by using them sequentially or in combination depending on the polarity of the solvents.

In the present invention, the solvent can be removed by filtering, concentrating, or drying the prepared extract or the fraction obtained by performing the fractionation process, and the filtration, concentration, and drying can all be performed. Specifically, the filtration can be performed using a filter paper or a pressure reducing filter, the concentration can be performed by pressure reducing concentration using a pressure reducing concentrator, etc., and the drying can be performed by a freeze-drying method.

In addition, additional purified fractions can be obtained by purifying the above-mentioned manufactured extract or the fraction obtained by performing the above-mentioned fractionation process using various chromatography methods such as silica gel column chromatography, thin layer chromatography, or high performance liquid chromatography.

The mixed extract of the extract of Cheonmun-dong, the extract of Rehmannia glutinosa and the extract of Baekbokryeong of the present invention is preferably a mixture of the extract of Cheonmun-dong, the extract of Rehmannia glutinosa and the extract of Baekbokryeong in a weight ratio of 1:1 to 5:1 to 5. In particular, it is more preferable that the extract of Cheonmun-dong, the extract of Rehmannia glutinosa and the extract of Baekbokryeong are mixed in a weight ratio of 1:1:1.

The composition of the present invention can be used for manufacturing a pharmaceutical formulation.

The above pharmaceutical formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granules, tablets or capsules.

In addition, the composition may additionally contain one or more active ingredients exhibiting the same or similar functions. For example, it may include known skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation components. If additional skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation components are included, the skin regeneration, wrinkle improvement, elasticity enhancement, anti-inflammation, antioxidant and/or anti-glycation improvement effects of the composition of the present invention may be further enhanced. When adding the above components, skin safety according to complex use, ease of formulation, and stability of the active ingredients may be considered. In one specific embodiment of the present invention, the composition may further include one or more components selected from the group consisting of skin regeneration components known in the art, such as retinoic acid, TGF, protein derived from animal placenta, betulinic acid and chlorella extract, antioxidant components known in the art, such as tocopherol, selenium, vitamin C and phenolic compounds, whitening components known in the art, substances that inhibit tyrosinase enzyme activity, such as kojic acid and arbutin, hydroquinone, vitamin-C (L-Ascorbic acid) and derivatives thereof, and various plant extracts. The additional component may be included in an amount of 0.0001 wt% to 10 wt% with respect to the total weight of the composition, and the content range may be adjusted according to requirements, such as skin safety and ease of formulation of the effective ingredient.

Additionally, the composition of the present invention may further comprise a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain a number of ingredients such as buffers, sterile water for injection, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.

The composition of the present invention can be administered orally or parenterally, and can be administered in the form of general pharmaceutical preparations, for example, in various oral and parenteral formulations for clinical administration. When formulated, it can be prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants that are commonly used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations can be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions, emulsions, and syrups, and in addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives may be included.

Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin.

In the present invention, the term 'whitening effect' refers to not only brightening the skin tone by inhibiting the synthesis of melanin pigment, but also improving skin hyperpigmentation such as blemishes or freckles caused by ultraviolet rays, hormones, or genetics.

In the present invention, the 'wrinkle improvement effect' refers to suppressing or inhibiting the formation of wrinkles on the skin or alleviating wrinkles that have already formed.

In the present invention, the 'elasticity-promoting effect' means increasing the elasticity of the skin, suppressing or inhibiting the loss of skin elasticity, or alleviating elasticity that has already been reduced.

In the present invention, the 'anti-inflammatory effect' refers to suppressing inflammation. The inflammation is one of the defense responses of living tissues to a certain stimulus, and refers to a complex lesion that causes three things: tissue degeneration, circulatory disorder, exudation, and tissue proliferation. More specifically, inflammation is a part of innate immunity, and like other animals, human innate immunity recognizes patterns of cell surfaces that exist specifically in pathogens. Phagocytes recognize cells with such surfaces as non-self and attack pathogens. If pathogens break through the physical barrier of the body and enter, an inflammatory response occurs. The inflammatory response is a non-specific defense action that creates a hostile environment for microorganisms that have invaded the wound site. In the inflammatory response, when a wound occurs or an external infectious agent enters the body, white blood cells responsible for the initial immune response rush in and express cytokines. Therefore, the amount of cytokine expression in cells is an indicator of the activation of the inflammatory response. Examples of skin diseases related to inflammation include atopic dermatitis, psoriasis, erythematous diseases caused by radiation, chemicals, or burns; Itching due to mountain ash, bullous dermatoses, lichen planus-like diseases or allergies; inflammatory hair loss such as seborrheic dermatitis, acne rosacea, pemphigus vulgaris, erythema multiforme exudative, erythema nodosum, balanitis, vulvitis or alopecia areata; cutaneous T-cell lymphomas, etc., but are not limited to these.

In the present invention, the term 'antioxidant effect' refers to the inhibition of oxidation of cells by highly reactive free radicals or reactive oxygen species (ROS) due to oxidative stress caused by intracellular metabolism or the influence of ultraviolet rays, and includes the reduction of damage to cells caused by removing free radicals or reactive oxygen species.

In the present invention, the term 'anti-glycation effect' means preventing or inhibiting glycation, a phenomenon in which glucose and proteins present in cells react and protein function in the body deteriorates.

In the present invention, the term "effective amount" means the amount of an extract that can exhibit a damaged whitening effect, improve wrinkles, increase elasticity, suppress inflammation, inhibit or alleviate cell oxidation, or inhibit the production of glycation. When the composition of the present invention contains an effective amount of a mixed extract of the Cheonmun-dong extract, the Rehmannia glutinosa extract, and the Poria cocos extract, it can provide desirable skin whitening effects, wrinkle improvement effects, elasticity promotion effects, anti-inflammatory effects, antioxidant effects, and anti-glycation effects. The effective amount of the mixed extract of the Cheonmun-dong extract, the Rehmannia glutinosa extract, and the Poria cocos extract included in the composition of the present invention will vary depending on the form in which the composition is manufactured, the method by which the compound is applied to the skin, and the time it remains on the skin. For example, when the composition is manufactured as a pharmaceutical formulation, it can contain the effective ingredient at a higher concentration than when it is manufactured as a cosmetic that is routinely applied to the skin. Therefore, the daily dosage is 0.1 to 100 mg/kg, preferably 30 to 80 mg/kg, and more preferably 50 to 60 mg/kg, based on the amount of the mixed extract of the above Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract, and can be administered 1 to 6 times a day.

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

The pharmaceutical formulation of the present invention may include a skin topical formulation.

When the mixed extract of the above Cheonmun-dong extract, Rehmannia glutinosa extract and Baekbokryeong extract is used as a skin external agent, it may additionally contain adjuvants commonly used in the field of dermatology, such as fatty substances, organic solvents, solubilizers, thickening and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in skin external agents. In addition, the above ingredients may be introduced in amounts commonly used in the field of dermatology.

When the mixed extract of the above Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract is provided in the form of a skin external preparation, it may have a formulation such as, but not limited to, an ointment, a patch, a gel, a cream, or a spray.

In addition, the skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, anti-oxidation and/or anti-glycation composition of the present invention can be used for manufacturing a cosmetic formulation.

The above cosmetic formulation may be in the form of a general emulsified formulation and a solubilized formulation. For example, it may have a formulation of a toner such as a flexible toner or a nourishing toner, an emulsion such as a facial lotion or a body lotion, a cream such as a nourishing cream, a moisture cream, an eye cream, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation in the form of a liquid type, a solid type or a spray type, a makeup remover such as a powder, a cleansing cream, a cleansing lotion, a cleansing oil, a cleanser such as a cleansing foam, a soap, a body wash, etc.

In addition, the cosmetic may contain, in addition to the mixed extract of the extract of Cheonmun-dong, the extract of Sukjihwang and the extract of Baekbokryeong, excipients commonly used in the field of cosmetics, such as fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics.

The above cosmetic formulation may contain a relatively high concentration of the active ingredient in the case of wash-off type cosmetics such as makeup removers and cleansers, in which the active ingredient remains on the skin for a short period of time. On the other hand, in the case of leave-on type cosmetics such as toners, emulsions, creams, and essences, in which the active ingredient remains on the skin for a long period of time, the active ingredient may be contained in a lower concentration than in wash-off type cosmetics. Although not limited thereto, in one specific embodiment of the present invention, the composition may contain the active ingredient in an amount of 0.0001 wt% to 10 wt% (preferably 0.0001 wt% to 1 wt%) based on the total weight of the composition. If the composition of the present invention contains less than 0.0001 wt% of the mixed extract of the extract of Cheonmun-dong, the extract of Rehmannia glutinosa, and the extract of Baekbokryeong, sufficient skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, anti-oxidation, and/or anti-glycation effects cannot be expected, and if it contains more than 10 wt%, unwanted reactions such as allergies may occur or problems with skin safety may arise, so this is to prevent this.

In addition, the skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammation, anti-oxidation and/or anti-glycation composition of the present invention can be used for manufacturing a food formulation.

The above food formulation refers to a food manufactured by adding a mixed extract of the above Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract to food materials such as beverages, teas, spices, gum, and confectionery, or by manufacturing the food in the form of encapsulation, powder, suspension, etc.

The above food formulation is very useful because it can be consumed on a daily basis and can be expected to have high skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, antioxidant and/or anti-glycation effects.

When the above-mentioned effective ingredient is used as a food additive, the mixed extract of the above-mentioned Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract can be added as it is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The mixed amount of the effective ingredient can be appropriately determined depending on its intended use (prevention, health, or therapeutic treatment). Generally, when manufacturing food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount can be below the above range, and since there is no problem in terms of safety, the effective ingredient can also be used in an amount greater than the above range.

There are no special restrictions on the types of the above foods. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the conventional sense.

When the food formulation is a beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, just like a typical beverage. The natural carbohydrates mentioned above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As a sweetener, a natural sweetener such as thaumatin and stevia extract, or a synthetic sweetener such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

In addition to the above, the food formulation may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the food formulation may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in mixtures. The proportion of these additives is not particularly important, 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.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are only intended to illustrate the present invention, and the content of the present invention is not limited to the following examples.

Manufacturing Example 1: Manufacturing of Cheonmundong extract

After thoroughly drying and cutting the Cheonmun-dong, 100 g of the dry weight was placed in a flask and extracted by steeping for 3 days with 1,000 g of extraction solvent (distilled water). The steeped extract was filtered through a filter with a pore size of 0.2 μm to prepare the Cheonmun-dong extract.

Manufacturing Example 2: Manufacturing of Rehmannia glutinosa extract

After thoroughly drying and cutting the Sukjihwang (Sukjidunhwang), 100g of the dry weight was placed in a flask and extracted by quenching with 1000g of extraction solvent (distilled water) for 3 days. The quenched extract was filtered through a filter with a pore size of 0.2㎛ to produce the Sukjihwang extract.

Manufacturing example 3: Baekbokryeong Preparation of extracts

After thoroughly drying and cutting the Poria cocos, 100 g of the dried weight was placed in a flask and extracted by quenching with 1,000 g of extraction solvent (distilled water) for 3 days. The quenched extract was filtered through a filter with a pore size of 0.2 μm to produce the Poria cocos extract.

Manufacturing example 4: Cheonmun-dong , Rehmannia glutinosa and Baekbokryeong's Preparation of mixed extracts

10 g of the extract of Cheonmun-dong of Manufacturing Example 1, 10 g of the extract of Sukjihwang of Manufacturing Example 2, and 10 g of the extract of Baekbokryeong of Manufacturing Example 3 were mixed to prepare a mixed extract of Cheonmun-dong, Sukjihwang, and Baekbokryeong.

Example 1: Inhibitory effect on melanin production

To confirm the whitening effect through inhibition of melanin production, the extract was added to the culture medium of mouse melanoma cells (B-16 mouse melanoma cells) and the total amount of melanin was measured according to the method described by Lotan R. et al. (Cancer Res. 40:3345-3350, 1980). In the experiment, toxicity to mouse melanoma cells was first evaluated and whitening evaluation was performed at a non-toxic concentration. DMSO was used as a negative control and arbutin was used as a positive control.

Specifically, the sample was added to the medium so that the final concentration was 100 ppm, and arbutin was added to the medium so that the final concentration was 100 ppm, and then melanoma cells were cultured for 3 days. Thereafter, the cells were detached from the culture vessel by trypsin treatment, centrifuged, and melanin was extracted. The detached cells were boiled in 1 ml of sodium hydroxide solution (1N concentration) for 10 minutes to dissolve the melanin, and the amount of melanin produced was measured by measuring the absorbance at 400 nm using a spectrophotometer.

The above melanin amount was measured by the method of expressing it as absorbance per unit cell number (1×10 ⁶ cells), and the total melanin amount relative to the control group was calculated as the inhibition rate (%), and the results are shown in Table 1 below. Each experiment was performed three times and expressed as the average value.

Effect of concentration on reduction of total melanin amount at cellular level (repetition = 3) Sample Melanin production (abs) Inhibition rate (%) Control (DMSO, 10ppm) 0.33 - Positive control (Arbutin, 100ppm) 0.228 30.91 Manufacturing Example 1 (100 ppm) 0.285 13.64 Manufacturing Example 2 (100 ppm) 0.290 12.12 Manufacturing Example 3 (100 ppm) 0.291 11.82 Manufacturing Example 4 (100 ppm) 0.237 28.18

As can be seen from the results in Table 1 above, the mixed extract of Manufacturing Example 4 showed an excellent effect of reducing the total amount of melanin even at a low concentration, indicating that it can be used for whitening purposes.

Example 2: Collagen synthesis promotion effect

The sample was added to the culture medium of human-derived fibroblasts to confirm the effect of promoting type I collagen synthesis at the cellular level. The synthesized collagen was quantitatively measured using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT).

The sample was added to the culture medium (DMEM medium) of fibroblasts at a final concentration of 10 ppm and cultured for 48 hours. The culture medium was then taken and the degree of type 1 collagen synthesis at each concentration was measured using a spectrophotometer at 450 nm using a PICP EIA kit.

To compare the effects, the level of collagen synthesis was measured using the same method for the culture medium of fibroblasts that were not treated with the sample (negative control group) and for the sample to which vitamin C (positive control group) was added to a final concentration of 52.85 ㎍/ml.

The increase rate of collagen production was calculated as the ratio of the relative collagen production to the negative control group, and the results are shown in Table 2 below. The experiment was performed four times and presented as the average value.

Collagen synthesis promotion effect (repetition = 4) Sample Type 1 collagen production (ng/ml) Increase in collagen production (%) Voice control group 150.2 - Positive control (vitamin C) 246.8 64.3 Manufacturing Example 1 (10 ppm) 164.3 9.39 Manufacturing Example 2 (10 ppm) 194.5 29.49 Manufacturing Example 3 (10 ppm) 198.2 31.96 Manufacturing Example 4 (10 ppm) 243.1 61.85

As shown in Table 2 above, when the mixed extract of Manufacturing Example 4 was treated, collagen synthesis was increased, and a collagen synthesis effect similar to that of when vitamin C, which is generally known to induce collagen synthesis, was applied was exhibited.

Example 3: Elastase Active inhibitory effect

The activity inhibition effect of elastase, an enzyme that breaks down elastin, was confirmed as follows.

Elastase was derived from human white blood cells, and MeOSuc-Ala-Ala-Pro-Val-pNA, a synthetic substrate, was used as a substrate of elastase. A 100 mM Tris (pH 7.5) solution was used as the buffer solution. Elastase was finally used at 0.2 mU using the buffer solution. In addition, the synthetic substrate of elastase was diluted using a buffer solution to a final concentration of 0.5 mM after making a 100 mM solution using DMSO. At this time, the positive control was set to 10 ppm of quercetin, known as an elastase inhibitor. The elastase inhibition candidate was added to a final concentration of 10 ppm. The reaction was performed in a 96-well plate and reacted at room temperature for 20 minutes. The absorbance was measured at 405 nm at 1-minute intervals using a spectrophotometer, and the slope of the absorbance versus time was calculated to determine the enzyme activity. The elastase inhibition rate was calculated using the following mathematical formula 1.

[Mathematical Formula 1]

Elastase activity inhibition effect (repetition = 3) Sample Enzyme activity Inhibition rate (%) Manufacturing Example 1 (10 ppm) 6.4 38.46 Manufacturing Example 2 (10 ppm) 6.9 33.65 Manufacturing Example 3 (10 ppm) 4.3 58.65 Manufacturing Example 4 (10 ppm) 3.7 64.42 Positive control (Quercetin, 10ppm) 3.5 66.35 Control (DMSO, 20ppm) 10.4 -

As can be seen in Table 3 above, when the mixed extract of Manufacturing Example 4 was treated, the effect was less than that of the positive control group, but it was found to have excellent elastase activity inhibition. That is, the mixed extract exhibited excellent elastase activity inhibition effect. Therefore, it was found that the mixed extract can be used for the purpose of increasing elasticity and improving wrinkles.

Example 4: Anti-inflammatory effect

To confirm the anti-inflammatory effect and skin trouble improvement effect, a nitric oxide (NO) production inhibition test was conducted using the GRIESS method using RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are mouse macrophages, were passaged several times, placed into a 24-well plate at a density of 3 × 10 ⁵ cells per well, and cultured for 24 hours. Then, the cell medium containing the sample was replaced with a final concentration of 10 ppm. At this time, L-NMMA (L-NG-Monomethylarginine), an NO production inhibitor, was treated together as a positive control and cultured for 30 minutes, and 1 ㎍ of LPS (Lipopolysaccharide) was treated as a stimulus and cultured for 24 hours. The supernatant was taken at 100 ㎕ and transferred to a 96-well plate, and 100 ㎕ of GRIESS solution was added at a time, reacted at room temperature for 10 minutes, and the NO inhibition effect was determined by measuring the absorbance at 540 nm. The NO production inhibition rate (%) was calculated using the following mathematical formula 2 and shown in Table 4 below. Each experiment was performed three times and shown as the average value.

[Mathematical formula 2]

NO production inhibition rate (%) = {(absorbance of negative control group - absorbance of each extract) / absorbance of negative control group} x 100

NO production inhibition effect (repetition = 3) Sample NO production inhibition rate (%) Control (DMSO, 10 ppm) - L-NMMA (positive control, 10 ppm) 41.02 Manufacturing Example 1 (10 ppm) 27.63 Manufacturing Example 2 (10 ppm) 26.44 Manufacturing Example 3 (10 ppm) 28.71 Manufacturing Example 4 (10 ppm) 39.26

As can be seen in Table 4 above, the mixed extract of Manufacturing Example 4 has lower activity than L-NMMA, a representative anti-inflammatory pharmaceutical substance, but exhibits excellent activity as a natural substance, so that a synergistic effect can be expected as the anti-inflammatory effect plays a supplementary role in improving skin troubles.

Example 5: Antioxidant effect - Free radical scavenging rate

To confirm the antioxidant activity of the sample, free radical scavenging activity was measured. Free radical scavenging activity was measured using DPPH (Blois, Nature 181, 1190, 1958).

DPPH is a relatively stable free radical that exhibits maximum absorption at 517 nm when in a radical state and loses absorbance when the radical is eliminated. DPPH used was from Sigma.

First, a standard DPPH ethanol solution of 1.5 mM (0.06 mg/ml) was prepared. Then, a 1000 μg/ml (0.1%) vitamin C solution was prepared using triple-distilled water, and then serially diluted by 1/2 each to prepare vitamin C samples with concentrations of 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 3.91, 1.95, 0.98, and 0.49 μg/ml. In addition, diluted solutions of Preparation Examples 1 to 4 were prepared at the same concentration as vitamin C. Then, the sample and the standard DPPH solution were added at the same ratio, stirred well, and reacted at 37 °C for 30 minutes. The absorbance at 517 nm was measured using a microplate reader (BioTek EL-340) to obtain the IC ₅₀ , which is shown in Table 5 below.

IC ₅₀ (half maximal inhibitory concentration) is the concentration of a substance required to remove 50% of the free radicals of the unadded control (DPPH in this case), and is a common way to express free radical scavenging activity.

Free radical scavenging effect (repetition number = 3) Sample IC ₅₀ (%w/v) Positive control (vitamin C) 0.0005 Manufacturing example 1 0.0016 Manufacturing example 2 0.0023 Manufacturing example 3 0.0019 Manufacturing example 4 0.0008

As shown in Table 5 above, the mixed extract of Manufacturing Example 4 had excellent free radical scavenging activity compared to Manufacturing Examples 1 to 3, and was confirmed to have an antioxidant effect similar to that of the positive control, vitamin C.

Example 6: Anti-glycation effect

To confirm the anti-glycation efficacy, glycation inhibition activity was measured using L-arginine and glucose.

First, 1 M L-arginine and 1 M glucose were dissolved in 1 M phosphate buffer (pH 7.4) to prepare, and the sample was diluted to 50 ppm using 1 M phosphate buffer to prepare. 1 M L-arginine and 1 M phosphate buffer were mixed in a ratio of 1 to 4, and 80 μl was dispensed into a 96-well plate. To this, 100 μl of the sample diluted to 50 ppm and 0.01 M aminoguanidin to be used as a positive control were added. After mixing these samples well, glucose diluted with 1 M phosphate buffer to a final glucose concentration of 0.1 M was added, and the reaction was performed at 70 °C for 4 hours. The degree of glycation was measured by measuring the absorbance at 420 nm using a spectrophotometer in a 96-well plate.

The Glycation experimental group in the following mathematical formula 1 is an experimental group in which glycation was induced by adding 1 M L-arginine and 1 M glucose, and in order to measure the absorbance of the sample itself, only 1 M L-arginine and the sample were added without adding glucose, and the absorbance was measured at 420 nm.

The glycation inhibition activity can be obtained using the following mathematical formula 3. The experiments were performed three times each and the average value was expressed.

[Mathematical Formula 3]

Sample (repeat =3) Glycosylation inhibition rate (%) Control (DMSO, 50 ppm) - Positive control (Aminoguanidine, 55 ppm) 54.89 Manufacturing Example 1 (50 ppm) 33.11 Manufacturing Example 2 (50 ppm) 49.17 Manufacturing Example 3 (50 ppm) 38.69 Manufacturing Example 4 (50 ppm) 69.97

As can be seen from the results in Table 6 above, the mixed extract of Manufacturing Example 4 according to the present invention can be seen to have a more excellent antiglycation effect than Aminoguanidine, which is known as an antiglycation substance. In other words, the mixed extract exhibits an excellent antiglycation effect, so that the antiglycation effect can play a supplementary role in whitening or improving wrinkles, and thus a synergistic effect can be expected.

Example of preparation 1: Manufacturing of pharmaceutical preparations

1. Preparation of tablets

0.2 mg of mixed extract of manufacturing example 4

Corn starch 100 mg

100 mg of lactose

Magnesium stearate 2 mg

After mixing the above ingredients, tablets were manufactured by pressing them according to the usual method for manufacturing tablets.

Example of preparation 2: Manufacturing of cosmetics

1. Preparation of nourishing cream

A nutritional cream containing the mixed extract of Manufacturing Example 4 as an effective ingredient was manufactured according to a conventional method, as shown in the composition below.

0.2 wt% of mixed extract of manufacturing example 4

Beta-1,3-glucan 5.0 wt%

Beeswax 10.0 wt%

Polysorbate 60 1.5 wt%

PEG 60 Hardened Castor Oil 2.0 wt%

Sorbitan sesquioleate 0.5 wt%

Liquid paraffin 10.0 wt%

Squalane 5.0 wt%

Caprylic/Capric Triglyceride 5.0 wt%

5.0 wt% glycerin

Butylene glycol 3.0 wt%

Propylene glycol 3.0 wt%

Triethanolamine 0.2 wt%

Preservative 0.05 wt%

Pigment 0.05 wt%

Fragrance 0.05 wt%

Purified water to 100 wt%

Example of preparation 3: Manufacturing of external skin preparations

1. Preparation of ointment

An ointment containing the mixed extract of Manufacturing Example 4 as an effective ingredient was manufactured according to a conventional method, as shown in the following composition.

0.5 wt% of mixed extract of manufacturing example 4

Beta-1,3-glucan 10.0 wt%

Beeswax 10.0 wt%

Polysorbate 60 5.0 wt%

PEG 60 Hardened Castor Oil 2.0 wt%

Sorbitan sesquioleate 0.5 wt%

Vaseline 5.0 wt%

10.0 wt% liquid paraffin

Squalane 5.0 wt%

Shea butter 3.0 wt%

Caprylic/Capric Triglyceride 5.0 wt%

10.0 wt% glycerin

Propylene glycol 10.2 wt%

Triethanolamine 0.2 wt%

Preservative 0.05 wt%

Pigment 0.05 wt%

Fragrance 0.05 wt%

Purified water to 100 wt%

Claims (22)

A skin whitening composition for the manufacture of cosmetics or foods, comprising a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract as an effective ingredient.
In paragraph 1,
Cheonmundong extract is an extract obtained by extracting Cheonmundong with one or more selected from the group consisting of water and organic solvents.
The extract of Rehmannia glutinosa is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents.
The Baekbokryeong extract is a skin whitening composition obtained by extracting Baekbokryeong with one or more selected from the group consisting of water and organic solvents.
In the second paragraph,
A composition for skin whitening, wherein the organic solvent is a polar solvent including a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; a nonpolar solvent including ether, chloroform, benzene, hexane, or dichloromethane; or a mixed solvent thereof.
An external skin preparation for preventing or treating skin hyperpigmentation, containing as an active ingredient a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract, and Poria cocos extract.
A composition for improving skin elasticity or wrinkles, comprising a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract as an effective ingredient, and intended for the manufacture of cosmetics or foods.
In paragraph 5,
Cheonmundong extract is an extract obtained by extracting Cheonmundong with one or more selected from the group consisting of water and organic solvents.
The extract of Rehmannia glutinosa is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents.
The Baekbokryeong extract is a composition for improving skin elasticity or wrinkles, which is an extract obtained by extracting Baekbokryeong with one or more selected from the group consisting of water and organic solvents.
In paragraph 6,
A composition for improving skin elasticity or wrinkles, wherein the organic solvent is a polar solvent including a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; a nonpolar solvent including ether, chloroform, benzene, hexane, or dichloromethane; or a mixed solvent thereof.
delete An anti-inflammatory composition comprising a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract, and Poria cocos extract as an effective ingredient.
In Article 9,
Cheonmundong extract is an extract obtained by extracting Cheonmundong with one or more selected from the group consisting of water and organic solvents.
The extract of Rehmannia glutinosa is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents.
The Baekbokryeong extract is an anti-inflammatory composition obtained by extracting Baekbokryeong with one or more selected from the group consisting of water and organic solvents.
In Article 10,
An anti-inflammatory composition wherein the organic solvent is a polar solvent including a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; a nonpolar solvent including ether, chloroform, benzene, hexane, or dichloromethane; or a mixed solvent thereof.
In Article 9,
An anti-inflammatory composition, the composition being for the manufacture of an anti-inflammatory medicine, cosmetic or food.
An antioxidant composition for the manufacture of cosmetics or foods, comprising a mixed extract of Cheonmun-dong extract, Rehmannia glutinosa extract, and Baekbokryeong extract as an active ingredient.
In Article 13,
Cheonmundong extract is an extract obtained by extracting Cheonmundong with one or more selected from the group consisting of water and organic solvents.
The extract of Rehmannia glutinosa is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents.
The Baekbokryeong extract is an antioxidant composition obtained by extracting Baekbokryeong with one or more selected from the group consisting of water and organic solvents.
In Article 14,
An antioxidant composition wherein the organic solvent is a polar solvent including a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; a nonpolar solvent including ether, chloroform, benzene, hexane, or dichloromethane; or a mixed solvent thereof.
delete delete delete delete delete In paragraph 4,
Cheonmundong extract is an extract obtained by extracting Cheonmundong with one or more selected from the group consisting of water and organic solvents.
The extract of Rehmannia glutinosa is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents.
Poria cocos extract is an external skin preparation for preventing or treating skin hyperpigmentation, which is obtained by extracting Poria cocos with one or more selected from the group consisting of water and organic solvents.
In Article 21,
An external skin preparation for preventing or treating skin hyperpigmentation, wherein the organic solvent is a polar solvent including a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; a nonpolar solvent including ether, chloroform, benzene, hexane, or dichloromethane; or a mixed solvent thereof.