KR20230023267A - Cosmetic composition comprising mixed extrracts of pinus koraiensis wood and kenaf - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a mixed extract of Pinus koraiensis wood and kenaf (Hibiscus cannabinus). One aspect of the present invention provides the cosmetic composition comprising a mixture of a pine nut (Pinus koraiensis) wood extract and a kenaf (Hibiscus cannabinus) extract as an active ingredient. The cosmetic composition of the present invention exhibits antioxidant, skin whitening and anti-wrinkle effects.

Description

COSMETIC COMPOSITION COMPRISING MIXED EXTRACTS OF PINUS KORAIENSIS WOOD AND KENAF COSMETIC COMPOSITION COMPRISING MIXED EXTRACTS OF PINUS KORAIENSIS WOOD AND KENAF

The present invention relates to a cosmetic composition comprising a mixed extract of pine nut wood and kenaf (yangma).

Pine tree is a plant of the coniferous pine family, and its scientific name is Pinus koraiensis . When the tree is cut down, the heartwood is red, so it is called Hongsong (紅松). It is distributed in the Korean Peninsula, Northeast China, Far East Russia, Japan Honshu and Shikoku, and in Korea, it grows wild in most alpine areas. In Korea, 230,000 ha of pine nuts are planted nationwide, which is one of the country's major forest income resources.

However, unlike pine trees, which are similar species, and are used in various fields such as cosmetic materials and functional foods, pine nuts are currently used mainly for food and wood for forestry, so their utilization is not high.

In particular, it is difficult to supply and demand materials for pine trees because there are not many felling or thinning, whereas pine trees have the advantage of being able to easily supply materials through large amounts of felling or thinning. There is a need.

On the other hand, kenaf is an annual or biennial plant of the dicotyledonous malvaceae family (Malvaceae), and its scientific name is Hibiscus cannabinus . It is also called 'yangma' in Korea, and is native to Africa and India. The stem is erect, 1.5 to 5 meters high, and the stem is 1 to 2 centimeters thick. Leaves are alternate, palm-shaped, deeply split, and have long petioles. Initially, the reason for introducing kenaf to Korea was to use this leaf as a forage for livestock such as cattle. Flowers bloom in summer, and 2 or 3 flowers similar to Mugunghwa hang in the axils of the leaves. Flowers are 10 cm in diameter, yellowish white, and dark red in the middle. The calyx is divided into 5 pieces, the tip is sharp, and there are many projections on the surface. There are 5 petals, and the stamens run together like a barrel. The flower is a type of hibiscus and is presumed to be edible. The fruits are legumes, 1 cm long, egg-shaped, divided into 5 rooms, and each room contains 3 to 5 seeds. Seeds are grayish-brown and contain 20% fat. This fat is refined and used as edible or industrial oil.

Fiber with a length of about 22 cm obtained by fermenting kenaf after harvesting can be used as a substitute for hemp or jute. is not being produced.

That is, kenaf leaves have been tried to be used as forage, but the stem part, which occupies most of the aerial part of kenaf, has only been tried to be used as a mixture for bioplastics, but there is no actual commercial use, especially in cosmetics. never used in the field.

The above background art is possessed or acquired by the inventor in the process of deriving the disclosure of the present application, and cannot necessarily be said to be known art disclosed to the general public prior to the present application.

The present invention is to solve the above problems, and an object of the present invention is to provide a functional cosmetic composition using a mixed extract of pine nut wood and kenaf.

However, the problem to be solved by the present invention is not limited to those mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

One aspect of the present invention provides a cosmetic composition comprising a mixture of a pine tree ( Pinus koraiensis ) wood extract and a kenaf ( Hibiscus cannabinus ) extract as an active ingredient.

According to one embodiment, the weight ratio of the pine wood extract and the kenaf extract may be 1:10 to 10:1.

According to one embodiment, the mixture may be included in an amount of 0.1% to 50% by weight, based on the total amount of the cosmetic composition.

According to one embodiment, the extract is composed of water, lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, chloroform, 1,3-butylene glycol, n-hexane, diethyl ether, butyl acetate and dichloromethane It may be extracted by using at least one selected from the group as an extraction solvent.

According to one embodiment, the extraction may be performed at a temperature of 1 °C to 40 °C for 4 hours to 60 hours.

According to one embodiment, the cosmetic composition contains 1,1-diphenyl-2-picrylhydrazyl (1,1-diphenyl-2-picrylhydrazyl, DPPH) radical scavenging activity, 2,2'-azinobis-(3 -Ethylbenzothiazoline-6-sulfonic acid) (2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid, ABTS) may have radical scavenging activity or SOD (superoxide dismutase)-like activity.

According to one embodiment, the cosmetic composition inhibits MITF (microphthalmia associated transcription factor) expression, TRP-1 (tyrosinase related protein-1) expression, TRP-2 (tyrosinase related protein-2) expression inhibition, tyrosinase (Tyrosinase) activity inhibition and elastase (elastase) activity may have at least one or more performance.

According to one embodiment, the cosmetic composition, pine nut ( Pinus koraiensis ) It may be to further include a leaf ethanol extract as an active ingredient.

According to one embodiment, with respect to 100 parts by weight of the mixture, the pine nut leaf ethanol extract may be included in 10 parts by weight to 100 parts by weight.

According to one embodiment, the cosmetic composition may further include, as an active ingredient, one or more vegetable extracts selected from the group consisting of centella asiatica extract, propolis extract, houttuynia cordata extract, green tea leaf extract, walnut extract, and wolfberry extract. .

According to one embodiment, based on 100 parts by weight of the mixture, the vegetable extract may be included in 1 part by weight to 30 parts by weight.

According to one embodiment, the cosmetic composition may be for antioxidant, whitening, wrinkle improvement, convergence, pore contraction, or inflammation inhibition.

According to one embodiment, the cosmetic is any one of basic cosmetics selected from the group consisting of lotion, toner, skin, lotion, emulsion, cream, essence, serum, gel, and pack, lipstick, tint, lip glow, lip balm, Cosmetics for washing, shampoo, At least one selected from the group consisting of cosmetics for hair, which is any one selected from the group consisting of rinse and treatment, and cosmetics for body, which is any one selected from the group consisting of body scrub, body lotion, body cream, and body wash It may contain.

The cosmetic composition according to the present invention, by containing a mixture of pine wood extract and kenaf extract, has the effect of exhibiting antioxidant, whitening and wrinkle improvement performance.

In particular, by simultaneously mixing the pine nut wood extract and the kenaf extract, it has an effect of exhibiting superior performance compared to the case where the pine wood extract or the kenaf extract is separately contained.

In addition, by including the pine nut leaf extract or vegetable extract at a certain component ratio, there is an effect of improving wrinkle improvement and anti-inflammatory performance.

Furthermore, by developing new products using pine nut, a forest resource that is currently not used as a material for industrial products, and kenaf, an agricultural product, it can contribute to the creation of new income for farmers, forestry workers and local companies.

1 is a result of measuring the electron donating ability of a cypress wood extract, a kenaf extract, and a mixture thereof.
2 is a result of measuring the ABTS radical scavenging activity of a pine nut extract, a kenaf extract, and a mixture thereof.
3 is a result of measuring SOD-like activities of pine nut extract, kenaf extract, and mixtures thereof.
4 is a result of measuring Tyrosinase inhibition rate of pine wood extract, kenaf extract, and mixtures thereof.
5 is a result of measuring the Elastase inhibition rate of pine wood extract, kenaf extract, and mixtures thereof.
6 is a result of measuring skin astringent activity of pine wood extract, kenaf extract, and mixtures thereof.
Figure 7 shows the cell viability of a mixture of pine wood extract and kenaf aerial part extract according to concentration in melanoma cells (B16F10).
Figure 8 shows the expression levels of MITF, tyrosinase, TRP-1 and TRP-2 proteins in melanoma cells (B16F10) by concentrations of a mixture of pine wood extract and kenaf aerial part extract.
9 shows the expression levels of MITF, TRP-1, TRP-2, and tyrosinase mRNA in melanoma cells (B16F10) by concentrations of a mixture of pine wood extract and kenaf aerial part extract.

Hereinafter, embodiments will be described in detail with reference to the accompanying drawings. However, since various changes can be made to the embodiments, the scope of the patent application is not limited or limited by these embodiments. It should be understood that all changes, equivalents or substitutes to the embodiments are included within the scope of rights.

Terms used in the examples are used only for descriptive purposes and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "include" or "have" are intended to designate that there is a feature, number, step, operation, component, part, or combination thereof described in the specification, but one or more other features It should be understood that the presence or addition of numbers, steps, operations, components, parts, or combinations thereof is not precluded.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiment belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and unless explicitly defined in the present application, they should not be interpreted in an ideal or excessively formal meaning. don't

In addition, in the description with reference to the accompanying drawings, the same reference numerals are given to the same components regardless of reference numerals, and overlapping descriptions thereof will be omitted. In describing the embodiment, if it is determined that a detailed description of a related known technology may unnecessarily obscure the gist of the embodiment, the detailed description will be omitted.

In addition, in describing the components of the embodiment, terms such as first, second, A, B, (a), and (b) may be used. These terms are only used to distinguish the component from other components, and the nature, order, or order of the corresponding component is not limited by the term. When an element is described as being “connected,” “coupled to,” or “connected” to another element, that element may be directly connected or connected to the other element, but there may be another element between the elements. It should be understood that may be "connected", "coupled" or "connected".

Components included in one embodiment and components having common functions will be described using the same names in other embodiments. Unless stated to the contrary, descriptions described in one embodiment may be applied to other embodiments, and detailed descriptions will be omitted to the extent of overlap.

One aspect of the present invention provides a cosmetic composition comprising a mixture of a pine tree ( Pinus koraiensis ) wood extract and a kenaf ( Hibiscus cannabinus ) extract as an active ingredient.

The cosmetic composition according to the present invention, by containing a mixture of pine wood extract and kenaf extract, has antioxidant, whitening and anti-wrinkle effects.

In particular, by simultaneously mixing the pine nut wood extract and the kenaf extract, it has an effect of exhibiting superior performance compared to the case where the pine wood extract or the kenaf extract is separately contained.

In the present invention, the pine nut wood extract and the kenaf extract include both extracts of pine nut wood or kenaf, fractions thereof, or active fractions separated from the fractions.

The pine tree wood is characterized by clear growth rings as the neck of the pine tree, heartwood is yellowish red, and sapwood is pale reddish yellowish white, so that the heartwood and sapwood are clearly distinguished.

Pinitol, pinosylbin, chrysin, pinobaxin and pinocembrin are contained in pine wood.

Pinitol is known as a substance that helps improve insulin resistance by supplementing chiro-inositol, which plays a role in blood sugar control, and also helps suppress aging.

Pinosylvin inhibits the production of various inflammation-inducing mediators by blocking the activation of NF-kB, an inflammation-inducing signal, and is a substance that can be usefully used in inflammatory diseases such as atopy, psoriasis, and acne.

Chrysin blocks the conversion of testosterone to estrogen. In addition, pinobanksin and pinocembrin are known as components exhibiting antifungal efficacy.

The cosmetic composition according to the present invention may exhibit anti-aging and anti-inflammatory effects by including a pine wood extract in which pinitol, pinosylbin, chrysin, pinovaxin and pinocembrin components contained in pine wood are effectively extracted. there is.

According to one embodiment, the weight ratio of the pine wood extract and the kenaf extract may be 1:10 to 10:1.

Preferably, the weight ratio may be 1:5 to 5:1, more preferably, 1:3 to 3:1, and even more preferably, 1:2 to 2:1 it could be

The weight ratio range is a range that can maximize the functions of the pine wood extract and the kenaf extract, and is a range that can maximize the efficacy compared to a single extract.

According to one embodiment, the mixture may be included in an amount of 0.1% to 50% by weight, based on the total amount of the cosmetic composition.

Preferably, of the total cosmetic composition, it may be included in 0.3% to 30% by weight, more preferably, it may be included in 0.5% to 20% by weight, and even more preferably, 1% to 10% by weight may be included in weight percent.

When the content of the mixture is less than the above range, effects such as antioxidant, whitening, wrinkle improvement, convergence, pore contraction, and inflammation inhibition may be insignificant.

On the other hand, when the above range is exceeded, formulation stability may be reduced or skin irritation may be increased, and may be uneconomical.

According to one embodiment, the extract is composed of water, lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, chloroform, 1,3-butylene glycol, n-hexane, diethyl ether, butyl acetate and dichloromethane It may be extracted by using at least one selected from the group as an extraction solvent.

Preferably, the extract may be a lower alcohol extract having 1 to 4 carbon atoms, and more preferably, an ethanol extract.

Ethanol as the extraction solvent may use 50% (v/v) to 90% (v/v) ethanol, preferably 60% (v/v) to 80% (v/v) ethanol. can

The concentration range of the ethanol may be within a range capable of maximizing the extraction efficiency of the active ingredient in the ethanol of the pine wood extract and the kenaf extract.

According to one embodiment, the extraction may be performed at a temperature of 1 °C to 40 °C for 4 hours to 60 hours. Preferably, it may be carried out for 24 hours to 48 hours at a temperature of 20 ℃ to 30 ℃.

If the temperature and time are out of range, the active ingredient in the extract may not be sufficiently extracted or the active ingredient may be destroyed.

According to one embodiment, the extraction may be performed 1 to 5 times. This is to ensure sufficient extraction of the active ingredient in the extract.

The extract may be prepared in a powder form by additionally performing filtration, concentration, and freeze-drying processes.

According to one embodiment, the cosmetic composition contains 1,1-diphenyl-2-picrylhydrazyl (1,1-diphenyl-2-picrylhydrazyl, DPPH) radical scavenging activity, 2,2'-azinobis-(3 -Ethylbenzothiazoline-6-sulfonic acid) (2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid, ABTS) may have radical scavenging activity or SOD (superoxide dismutase)-like activity.

According to one embodiment, the cosmetic composition inhibits MITF (microphthalmia associated transcription factor) expression, TRP-1 (tyrosinase related protein-1) expression, TRP-2 (tyrosinase related protein-2) expression inhibition, tyrosinase (Tyrosinase) activity inhibition and elastase (elastase) activity may have at least one or more performance.

In general, melanin is produced to protect the skin from external stimuli such as ultraviolet rays. Excessive deposition of melanin in the skin can result in unwanted freckles, senile spots, liver spots, melasma, brown or sunspots, solar pigment spots, wounds including scratches and burns, or post-inflammatory hyperpigmentation due to dermatitis, phototoxic reactions, or other similar small It can cause fixed pigmented lesions.

Melanin production is regulated by three enzymes: tyrosinase, TRP-1 (tyrosinase related protein-1), and TRP-2. The biosynthetic mechanism of melanin is that tyrosine, the starting material, is transformed into DOPA (3,4-dihydroxyphenylalanin) and DOPA quinone by the rate-limiting enzyme tyrosinase, and the intermediate 5,6-dihydroxyindole- It is oxidized to 2-carboxylic acid to produce eumelanin. In addition, when keratinocytes of the skin are stimulated by ultraviolet rays, signal transmitters such as α-MSH (α-melanocyte sitmulating hormone) and ACTH (adrenocorricotropic hormone) are secreted into melanocytes. MC1R (melanocortin type 1 receptor) receptors in melanocytes accept these stimuli and increase intracellular cAMP (cyclic adenosine monophosphate) levels, and the increase in cAMP causes activation of PKA (protein kinase A), followed by CREB (cAMP response). element-binding protein) increases phosphorylation. Phosphorylated CREB causes the expression of MITF (microphthalmia associated transcription factor), a specific transcription factor of melanocytes. The expression of MITF eventually promotes binding to the promoters of tyrosinase, TRP-1, and TRP-2, which are major enzymes for melanin production, and increases melanin biosynthesis.

As UV rays exposed to the skin increase, matrix metalloproteinases (MMPs), which degrade substrates among various proteolytic enzymes, play an important role in the occurrence of skin photoaging. Extracellular matrix proteins in the skin dermis play an important role in maintaining the structure and elasticity of the skin, of which 80 to 85% is composed of type I procollagen. Type I procollagen is synthesized in the form of procollagen, which is a precursor, and then released out of the cell to make finished collagen after amino and carboxyl ends are cut off. Reconstitution of extracellular matrix proteins in the skin dermis involves several biological reactions, among which MMPs play an important role in degrading matrix proteins, and intestitial collagenase (MMP-1), 72kD gelatinase (MMP-2), and 92kD gelatinase ( MMP-9), neutrophil collagenase (MMP-8), collagenase (MMP-13), and more than 26 MMPs are already known. The expression of MMPs increases in the skin exposed to UV rays, and the increased MMPs degrade collagen constituting the skin, resulting in a deficiency of matrix protein, resulting in skin aging and wrinkles.

The cosmetic composition according to the present invention is a 1,1-diphenyl-2-picrylhydrazyl (1,1-diphenyl-2-picrylhydrazyl, DPPH) radical inhibition, 2,2'-azinobis- (3-ethylbenzo Thiazoline-6-sulfonic acid) (2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid, ABTS) radical inhibition, Thiobarbituric acid reactive substances (TBARs) inhibition, antioxidant protection By having an antioxidant protection factor (PF) increasing activity, it shows an antioxidant effect.

According to one embodiment, the cosmetic composition, pine nut ( Pinus koraiensis ) It may be to further include a leaf ethanol extract as an active ingredient.

The pine nut leaf ethanol extract maintains the form of elastin molecules by inhibiting esterase activity, activates Pro COL1A2, MMP-1 (Matrix Metalloproteinase-1) and MMP-9 (Matrix Metalloproteinase By suppressing the expression of -9), the anti-wrinkle effect can be improved.

In addition, by containing abundant polyphenol-based physiologically active substances, it is possible to exhibit a skin inflammation inhibitory effect.

The cosmetic composition according to the present invention is characterized in that anti-oxidation, whitening, anti-wrinkle and anti-inflammatory functions can be secured at the same time by including pine nut wood extract, kenaf extract and pine nut leaf extract at the same time.

According to one embodiment, with respect to 100 parts by weight of the mixture, the pine nut leaf ethanol extract may be included in 10 parts by weight to 100 parts by weight.

Preferably, with respect to 100 parts by weight of the mixture, the pine nut leaf ethanol extract may be included in 10 parts by weight to 80 parts by weight, more preferably, 30 parts by weight to 70 parts by weight.

The content range of the pine nut leaf ethanol extract is an optimal content range that can secure anti-wrinkle and anti-inflammatory functions at the same time without reducing the efficacy of the pine nut extract and the kenaf extract.

According to one embodiment, the cosmetic composition may further include, as an active ingredient, one or more vegetable extracts selected from the group consisting of centella asiatica extract, propolis extract, houttuynia cordata extract, green tea leaf extract, walnut extract, and wolfberry extract. .

According to one embodiment, based on 100 parts by weight of the mixture, the vegetable extract may be included in 1 part by weight to 30 parts by weight.

Preferably, based on 100 parts by weight of the mixture, the vegetable extract may be included in 1 part by weight to 20 parts by weight.

The vegetable extract can further enhance the antioxidant, whitening, and antifungal effects of the cosmetic composition according to the present invention, and provides other active ingredients not included in the pine nut extract, kenaf extract, or pine nut leaf extract at an appropriate level. , It can play a role in improving antioxidant, whitening, wrinkle improvement, convergence, pore contraction, and anti-inflammatory function.

Centella Asiatica is a perennial plant belonging to the Apiaceae family and contains Asiatic acid, Asiaticoside, and Madecassic acid as main components. It is effective for prevention, anti-aging, and skin regeneration.

Propolis is a yellowish green, dark brown sticky substance that is applied around the beehive as one of the substances constituting the bee's nest, and the propolis extract has antibiotic, anti-inflammatory, and antioxidant effects.

Eoseongcho is a plant in the family of more than 300 and is called medicinal buckwheat. Its main components include quercetin, quercitrin, isoquercetin, afzerin, rutin as flavonoids, and aristolactam A, AII, B, BII, piperolactam A, norcepharadione B, cepharadione B, and splendidine as alkaloids. there is. The Eoseongcho extract is effective in antioxidant, anti-inflammatory, whitening, and skin regeneration.

Green tea leaves contain 5 to 8 times more vitamin C than Levon, so it is effective in suppressing freckles and freckles, soothing the skin and removing free radicals to prevent skin aging. It can enhance pore contraction and anti-inflammatory effects.

Liriopeplatyphylla Wang et Tang is a perennial herbaceous plant belonging to the Liliaceae family, and the extract of Liriopeplatyphylla Wang et Tang improves the anti-inflammatory effect.

It is a fruit of the goji berry tree ( Lycium chinense ) and contains various functional ingredients such as betaine, cholin, physalien, rutin, and β zeazanthin, and the goji berry extract can improve antioxidant and antibacterial effects.

According to one embodiment, the vegetable extract, Centella Asiatica extract 1 part by weight to 10 parts by weight, propolis extract 1 part by weight to 8 parts by weight, houttuynia cordata extract 1 part by weight to 5 parts by weight, green tea leaf extract 1 part by weight to 5 parts by weight Part by weight, it may include 1 part by weight to 3 parts by weight of the extract of Mammoth sinensis and 1 part by weight to 3 parts by weight of the goji berry extract.

The content range of each component included in the vegetable extract corresponds to a range that can minimize side effects that may occur due to interactions between each component while maximizing the function of the active ingredient in each extract.

According to one embodiment, the cosmetic composition may be for antioxidant, whitening, wrinkle improvement, convergence, pore contraction, or inflammation inhibition.

The cosmetic composition according to the present invention may further include conventional adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, etc., which are commonly used in cosmetic compositions, in addition to the above active ingredients. For example, the cosmetic composition may further include auxiliary ingredients such as glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, and allantoin.

In addition, oil and fat ingredients, humectants, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling agents , Antiperspirants, purified water and other ingredients may be included.

Examples of the oil and fat component include ester oils, hydrocarbon oils, silicone oils, fluorine oils, animal fats and vegetable oils.

Examples of ester oils include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid. Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, adipine Diisopropyl acid, isoalkyl neopentanoate, tri(capryl, capric acid) glyceryl, trimethylolpropane tri2-ethylhexanoate, trimethylolpropane triisostearate, pentaelislitol tetra2-ethylhexanoate , Cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinooleate, isostea laurate Lil, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, 2-ethylhexanoate Aryl, 2-ethyl stearyl hexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di(caprylic, capric acid) propylene glycol, propylene glycol dicaprylate, dicap Neopentyl glycol prinate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate , Octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleate, polyglycerin isostearate, Triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, dimalate Isostearyl, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, Cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phyteryl isostearate, phyteryl oleate, 12-stealoyl and esters such as isocetyl hydroxystearate, stearyl 12-stealoylhydroxystearate, and isostearyl 12-stealoylhydroxystearate.

Examples of hydrocarbon-based fats and oils include hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybuden, microcrystalline wax, and vaseline.

Examples of silicone oils include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane/methylcetyloxysiloxane copolymer, dimethylsiloxane/methylstealoxysiloxane copolymer, alkyl Modified silicone oil, amino modified silicone oil, etc. are mentioned.

Examples of fluorine-based fats and oils include perfluoropolyether and the like.

Animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, birdflower oil, soybean oil, corn oil, rapeseed oil, algae oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil , cottonseed oil, palm oil, kukui nut oil, wheat germ oil, rice germ oil, shea butter, moonshine colostrum, marker damian nut oil, meadowsweet oil, egg yolk oil, beef tallow, horse oil, mink oil, orange raffia oil, jojoba oil , animal or plant fats and oils such as candelilla wax, carnaba wax, liquid lanolin, and hydrogenated castor oil.

Examples of the moisturizer include water-soluble low-molecular moisturizers, fat-soluble molecular moisturizers, water-soluble polymers, and oil-soluble polymers.

Examples of water-soluble low-molecular moisturizers include serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n = 2 or more), polyglycerol B (polymerization degree n = 2 or more), lactic acid, lactate, etc. are mentioned.

Cholesterol, cholesterol ester, etc. are mentioned as a fat-soluble low-molecular moisturizer.

Examples of water-soluble polymers include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water-soluble chitin, chitosan, dextrin, and the like. can Examples of the fat-soluble polymer include polyvinylpyrrolidone/eicosene copolymer, polyvinylpyrrolidone/hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and high molecular silicone.

Examples of the emollient agent include long-chain acyl glutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester, and the like.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants.

Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE·POP (polyoxyethylene·polyoxypropylene) copolymer, POE·POP alkyl ether, polyether modified silicone, lauric acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Anionic surfactants include fatty acid soaps, alpha-acyl sulfonates, alkyl sulfonates, alkyl allyl sulfonates, alkyl naphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkyl amide sulfates, alkyl phosphates, POE alkyl phosphates, and alkyl amide phosphates. , alkyloylalkyl taurine salts, N-acylamino acid salts, POE alkyl ether carboxylate salts, alkyl sulfosuccinic acid salts, sodium alkyl sulfoacetate, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, and the like. .

As the cationic surfactant, alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, chloride Benzalkonium, stearic acid diethylaminoethylamide, stearic acid dimethylaminopropylamide, lanolin derivative quaternary ammonium salt, etc. are mentioned.

Amphoteric surfactants include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amide sulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amideamine type, etc. An amphoteric surfactant etc. are mentioned.

As organic and inorganic pigments, silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide , inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine and their complexes, polyamide, polyester, polypropylene, polystyrene, polyurethane , vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene styrene copolymer, silk powder, cellulose, CI pigment yellow, CI pigment organic pigments such as orange, composite pigments of these inorganic pigments and organic pigments, and the like are exemplified.

As the organic powder, metal soaps such as calcium stearate, zinc sodium cetylate, zinc laurylate, calcium laurylate, etc., metal alkylphosphates, N-lauroyl-beta-alanine calcium, N-lauroyl-beta- Acylamino acid polyvalent metal salts such as zinc alanine and calcium N-lauroylglycine, polyvalent metal salts of amide sulfonic acids such as calcium N-lauroyl-taurine and calcium N-palmitoyl-taurine, N-epsilon-lauroyl-L- N-acyl basic amino acids such as lysine, N-epsilon-palmitoylizine, N-alpha-paritoylolnithine, N-alpha-lauroylarginine, and N-alpha-hydrogenated beef tallow fatty acid acylarginine, N-lauroyl glycol N-acyl polypeptides such as lysylglycine, alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid, polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene/styrene copolymer A polymer, ethylene tetrafluoride, etc. are mentioned.

Examples of UV absorbers include para-aminobenzoic acid, ethyl para-aminobenzoate, amyl para-aminobenzoate, octyl para-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, and benzyl cinnamate. , paramethoxycinnamic acid-2-ethoxyethyl, paramethoxycinnamic acid octyl, diparamethoxycinnamic acid mono-2-ethylhexane glyceryl, paramethoxycinnamic acid isopropyl, diisopropyl-diisopropylcinnamic acid ester mixture, uro Cannic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and its salts, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone sodium disulfonate, dihydroxybenzophenone , tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1 ,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, etc. are mentioned.

Bactericides include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zincophylthione, benzalkonium chloride, photosensitizer So No. 301, mononitroguacol sodium, undecyrenic acid, etc. are mentioned.

Examples of antioxidants include butylhydroxyanisole, propyl gallic acid, and elisorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumalate, succinic acid, sodium succinate, sodium hydroxide, and sodium monohydrogenphosphate.

Examples of alcohol include higher alcohols such as cetyl alcohol.

The ingredients in the cosmetic composition are not limited thereto, and any of the above ingredients can be blended within a range that does not impair the object and effect of the present invention, but is preferably 0.01% to 5% by weight relative to the total weight, more preferably may be blended in an amount of 0.01% to 3% by weight.

The cosmetic composition according to the present invention is a solution, external ointment, cream, foam, nutrient lotion, softening lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid cleanser, bath additive, sunscreen cream, sun oil, Suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays may be prepared in formulations selected from the group consisting of It is not limited thereto.

According to one embodiment, the cosmetic is any one of basic cosmetics selected from the group consisting of lotion, toner, skin, lotion, emulsion, cream, essence, serum, gel, and pack, lipstick, tint, lip glow, lip balm, Cosmetics for washing, shampoo, At least one selected from the group consisting of cosmetics for hair, which is any one selected from the group consisting of rinse and treatment, and cosmetics for body, which is any one selected from the group consisting of body scrub, body lotion, body cream, and body wash It may contain.

Hereinafter, the present invention will be described in more detail by examples and comparative examples.

However, the following examples are only for illustrating the present invention, and the content of the present invention is not limited to the following examples.

<Example> Preparation of a cosmetic composition containing pine wood extract and kenaf extract

1) Preparation of a mixture of pine wood extract and kenaf extract

5 kg each of pine nut timber and kenaf aerial parts were collected, dried in a natural shade, and then pulverized. After adding 70% (v/v) of ethanol 10 times the weight of the sample to the pulverized sample, soaking at room temperature for about 24 hours, the supernatant and precipitate were separated and extracted three times in the same manner. Each sample extract was filtered using a filter paper (Whatman No. 2), concentrated under reduced pressure with an EYELA evaporator, and freeze-dried after removing the solvent, and the sample was stored at -20 ° C. Then, a mixture (sample solution) was prepared by mixing the freeze-dried pine wood extract and the aerial part extract of kenaf in a ratio of 1:1.

2) Preparation of cosmetic composition

A cosmetic composition was prepared by mixing the prepared mixture with a mixed solvent including purified water, ethanol, glycerin, and the like, and other additives.

<Experimental Example 1> Confirmation of antioxidant effect: Measurement of electron donating ability of pine nut wood and kenaf mixed extract

In order to confirm the antioxidant effect of the mixture of the above-prepared pine wood extract and the aerial part extract of Kenaf, the electron donating ability was measured. As a comparative example, pine wood extract and kenaf aerial part extract were used alone.

Electron donating abilities (EDA) were measured by modifying the method of Blois. After adding 60 μl of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 120 μl of each extract for each concentration, they were mixed and allowed to stand for 15 minutes. Then, the absorbance was measured at 517 nm using a microplate reader. The electron-donating ability is expressed as the rate of decrease in absorbance between the added and non-added portions of the sample solution.

Electron donating ability (%) = (1 - (absorbance of sample added group/absorbance of non-added group)) × 100

1 is a result of measuring the electron donating ability of a cypress wood extract, a kenaf extract, and a mixture thereof.

1A is the result of measuring the electron donating ability of the pine nut extract, FIG. 1B is the result of measuring the electron donating ability of the kenaf extract, and FIG. 1C is the result of measuring the electron donating ability of the mixture.

Referring to FIG. 1, as a result of measuring the electron donating ability of the pine wood extract, the kenaf stem extract, and a mixture thereof, it can be seen that the electron donating ability increases as the concentration increases. In addition, the pine tree extract and the mixture showed 91.6% and 93.7% of the effect at the highest concentration of 1,000 μg/ml, respectively, and the kenaf stem extract showed an effect of 38.2% at 100 μg/ml, but at 1,000 μg/ml, It was confirmed that the effect was 84.8%.

<Experimental Example 2> Confirmation of antioxidant effect: ABTS of pine tree wood and kenaf mixed extract ⁺ Verification of radical scavenging ability

In order to confirm the antioxidant effect of the mixture of the above-prepared pine tree extract and the aerial part extract of kenaf, ABTS ⁺ radical scavenging activity was confirmed. As a comparative example, pine wood extract and kenaf aerial part extract were used alone.

ABTS ⁺ radical scavenging activity assay is an in vitro test method that can measure antioxidant activity indirectly by replacing radicals generated in the human body, and the experimental principle is the same as DPPH assay.

After mixing 7 mM 2,2-azino-bis (3-ethyl-benthiazoline-6-sulfonic acid) and 2.45 mM potassium persulfate, leave it at room temperature for 24 hours to form ABTS ⁺ radical. 100ul of the solution diluted with ethanol. 100 μl of the extract for each concentration was mixed and the absorbance at 735 nm was measured.

ABTS radical scavenging ability was calculated by the following formula.

ABTS radical scavenging ability (%) = (1 - (absorbance of sample added group / absorbance of non-added group)) × 100

2 is a result of measuring the ABTS radical scavenging activity of a pine nut extract, a kenaf extract, and a mixture thereof.

FIG. 2A is the result of measuring the ABTS radical scavenging activity of the pine nut extract, FIG. 2B is the result of measuring the ABTS radical scavenging activity of the kenaf extract, and FIG. 2C is the result of measuring the ABTS radical scavenging activity of the mixture.

Referring to FIG. 2, as a result of measuring the ABTS scavenging ability of pine nut wood extract, kenaf stem extract, and mixtures thereof, pine nut wood extract showed 90.8% scavenging ability at a concentration of 1,000 μg/ml, and kenaf stem extract and mixture showed an effect of 98.8% and 99% at a concentration of 1,000 μg/ml, respectively, confirming that the ABTS scavenging ability was excellent.

<Experimental Example 3> Confirmation of antioxidant effect: Measurement of SOD-like activity of pine nut wood and kenaf mixed extract

In order to confirm the antioxidant effect of the mixture of the above-prepared pine wood extract and kenaf aerial part extract, 130 μl of Tris-HCl buffer (50 mM Tris-HCl buffer, pH 8.5) and 7.2 mM pyrogallol were added to 20 μl of sample solution for each concentration. After adding and mixing 20 μl, the mixture was reacted at 37° C. for 10 minutes. Thereafter, the absorbance at 420 nm of the amount of oxidized pyrogallol was measured using a microplate reader.

SOD-like activity was calculated by the following equation.

SOD-like activity (%) = (1 - (absorbance of sample added group / absorbance of non-added group)) × 100

3 is a result of measuring SOD-like activities of pine nut extract, kenaf extract, and mixtures thereof.

Fig. 3A is the measurement result of SOD-like activity of pine nut extract, Fig. 3B is the measurement result of SOD-like activity of kenaf extract, and Fig. 3C is the measurement result of SOD-like activity of the mixture.

Referring to FIG. 3, as a result of measuring the SOD-like activity of pine nut wood extract, kenaf stem extract, and mixtures thereof, the pine wood extract was 1.91% at 5 μg/ml concentration, 3.81% at 100 μg/ml concentration, and 1000 The activity of 10.99% at μg/ml and the kenaf stem extract were 1.52% at 5 μg/ml, 3.62% at 100 μg/ml, and 7.56% at 1000 μg/ml. It can be seen that the mixture exhibits an activity of 0.95% at a concentration of 5 μg/ml, 2.28% at a concentration of 100 μg/ml, and 3.23% at a concentration of 1000 μg/ml.

<Experimental Example 4> Confirmation of whitening effect: Tyrosinase inhibition rate measurement

Tyrosinase is an enzyme that plays a key role in the melanin synthesis inhibition mechanism. It oxidizes tyrosine in the presence of oxygen to produce melanin, and the whitening effect can be confirmed by measuring the tyrosinase inhibitory activity of the extract.

Tyrosinase inhibition rate of the above-prepared pine wood extract, kenaf aerial part extract, and mixtures thereof was measured according to the method of Yagi et al. To 40 μl of the sample solution, 80 μl of 67 mM sodium phosphate buffer (pH 6.8), 40 μl of 10 mM L-DOPA, and 40 μl of mushroom tyrosinase (200 U/ml) were added and reacted at 37 °C for 10 minutes. DOPA chrome was measured at 475 nm.

Tyrosinase inhibitory activity was expressed as the rate of decrease in absorbance between the sample solution added and unadded.

Tyrosinase inhibition rate (%) = (1 - (absorbance of sample added group / absorbance of non-added group)) × 100

4 is a result of measuring Tyrosinase inhibition rate of pine wood extract, kenaf extract, and mixtures thereof.

Figure 4A is the Tyrosinase measurement result of the pine wood extract, Figure 4B is the Tyrosinase inhibition rate measurement result of the kenaf extract, and Figure 4C is the Tyrosinase inhibition rate measurement result of the mixture.

Referring to FIG. 4, as a result of measuring the Tyrosinase inhibitory activity of pine nut wood extract, kenaf stem extract, and mixtures thereof, pine nut wood extract showed 40.9% activity at the highest concentration of 1,000 μg/ml, and kenaf stem extract At the highest concentration of 1,000 μg/ml, the activity was 27.4%, and the mixture showed 42.6% inhibitory activity at 1,000 μg/ml. That is, it was confirmed that the inhibitory activity increased as the concentration of all extracts increased.

<Experimental Example 5> Confirmation of wrinkle improvement effect: Elastase inhibition rate measurement

Elastase inhibition rate of the above-prepared pine wood extract, kenaf aerial part extract, and mixtures thereof was measured by modifying the Cannell method.

Using N-succinyl-(L-Ala)3-p-nitroanilide as a substrate, the amount of p-nitroanilide generated from the substrate was measured at 445 nm at 37°C for 30 minutes. That is, 40 μl of each test solution was prepared to a certain concentration, and 40 μl of a solution of porcine pancreas elastase (0.5 U/ml) dissolved in 50 mM tris-HCl buffer (pH 8.6) and 0.4 M tris-HCl buffer (pH 8.6) were added. 8.6) After adding 40 μl and heating at 37 ° C for 2 minutes, 80 μl of N-succinyl-(L-Ala)3-p-nitroanilide (3.2 mM) dissolved in 50 mM tris-HCl buffer (pH 8.6) was added as a substrate. and reacted for 30 minutes to measure.

Elastase inhibition rate was expressed as the rate of decrease in absorbance between the sample solution added and non-added.

Elastase inhibition rate (%) = (1 - (absorbance of sample added group / absorbance of non-added group)) × 100

5 is a result of measuring the Elastase inhibition rate of pine wood extract, kenaf extract, and mixtures thereof.

FIG. 5A is the result of measuring the elastase inhibition rate of the pine nut extract, FIG. 5B is the result of measuring the elastase inhibition rate of the kenaf extract, and FIG. 5C is the result of measuring the elastase inhibition rate of the mixture.

Referring to FIG. 5, the pine wood extract exhibited an activity of 0.58% at 10 μg/ml, 4.44% at 100 μg/ml, and 13.08% at 1,000 μg/ml, and the kenaf stem extract had an activity of 10 μg/ml. The activity was 2.8% at 100 μg/ml concentration, 6.83% at 100 μg/ml concentration, and 11.35% at 1,000 μg/ml. It can be confirmed that the extract exhibited an activity of 20.15% at ug/ml and exhibited higher inhibitory activity than that of a single extract.

<Experimental Example 6> Confirmation of convergence and pore contraction effect

The skin astringent activity of the above-prepared cypress wood extract, kenaf aerial part extract, and mixtures thereof was measured according to the method of Lee et al.

0.5 ml of a 0.5% henoglobin solution dissolved in 1X PBS was added to 0.5 ml of the sample solution prepared to a certain concentration, mixed with shaking for 1 minute, and then centrifuged at 1,500 rpm for 3 minutes. After taking 150 μl of the supernatant, the absorbance at 575 nm was measured. The skin astringent activity was expressed as the rate of decrease in absorbance between the sample solution added and non-added.

Skin astringent activity (%) = (1 - (absorbance of sample added group / absorbance of non-added group)) × 100

6 is a result of measuring skin astringent activity of pine wood extract, kenaf extract, and mixtures thereof.

Figure 6A is the result of measuring the skin astringent activity of the pine nut extract, Figure 6B is the result of measuring the skin astringent activity of the kenaf extract, and Figure 6C is the result of measuring the skin astringent activity of the mixture.

Referring to FIG. 6, the pine wood extract exhibited an astringent activity of 8.57% at a concentration of 50 μg/ml, 47.53% at a concentration of 500 μg/ml, and 68.4% at a concentration of 5,000 μg/ml, and the kenaf stem extract exhibited an astringent activity of 100 μg/ml. The astringent activity was 0.8% at ㎖ concentration, 12.4% at 500 ug/mL concentration, and 113.33% at 5,000 ug/mL concentration. , it was confirmed that the activity of 111.51% was exhibited at 5,000 μg/ml.

<Experimental Example 7> Cytotoxicity evaluation: cell viability measurement

1) Cell line and cell culture

B16F10, a melanoma cell used in this experiment, was purchased from ATCC (USA) and used. For the culture of B16F10 cells, DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin (100 U/ml) was used, and the cells were subcultured in a 37°C, 5% CO incubator.

2) Measurement of cell viability by MTT assay

To measure cell viability, 180 μl of melanoma cells (B16F10) were dispensed in a 96 well plate to be 1×105 cells/well, samples were prepared by concentration, 20 μl was added, and cultured for 24 hours in a 37 °C, 5% CO ₂ incubator. did After adding 40 μl of MTT solution prepared at a concentration of 2.5 mg/ml and incubating for 4 hours, the culture medium was removed, 100 μl of DMSO was added to each well, reacted at room temperature for 10 minutes, and absorbance was measured at 540 nm with an ELISA reader. did

Cell viability was measured by the decrease in absorbance of the sample solution added group and non-added group.

Cell viability (%) = (1 - (absorbance of sample added group / absorbance of non-added group)) x 100

Figure 7 shows the cell viability of a mixture of pine wood extract and kenaf aerial part extract according to concentration in melanoma cells (B16F10).

Referring to Figure 7, it can be seen that the mixture exhibits a cell viability of 97% at a concentration of 1000 μg/ml.

Therefore, the measurement of whitening-related signaling factors in the following melanoma cells was performed at concentrations of 25, 50, and 100 μg/ml, which are concentrations close to 100% viability.

<Experimental Example 8> Confirmation of whitening effect and anti-inflammatory effect: Protein expression measurement through Western Blot

In order to investigate the mRNA expression inhibitory effect of the mixture on MITF, TRP-1, TRP-2 and tyrosinase, which are factors related to melanin synthesis, B16F10 mouse melanoma cells were treated with 25, 50, and 100 μg/ml of the extract by concentration. The control was treated with α-MSH to overexpress melanin, and the normal part was set to a section where α-MSH was not treated. 24 hours later, mRNA expression was measured by RT-PCR, and at this time, GAPDH, a house keeping gene with little difference in expression level under various cell conditions, was used as a positive control.

In a specific way, in order to examine the activity of the whitening-related factors of MITF, TRP-1, TRP-2, and tyrosinase, the cell line (B16F10) was dispensed in a 100 mm tissue culture dish at 1×10 ⁶ cells/well and then 24 hours Cells were stabilized by culturing for a period of time. After removing the medium, α-MSH (100 nM) was treated for 2 hours, and the extract was incubated for 24 hours with a medium treated by concentration, and then the medium was removed again and washed twice with 1 × PBS. 100 μl of Complete mini 1 tab was dissolved in 10 ml of RIPA buffer and centrifuged at 4° C. and 13,200 rpm for 20 minutes. The supernatant obtained by centrifugation was quantified with a BCA protein assay kit, and 20 μl of protein was separated by electrophoresis on a 10% acrylamide gel. The separated protein was transferred to a polyvinylidene fluoride (PVDF) membrane using a transfer device and then incubated in blocking buffer (5% skim milk in TBST) for 1 hour at room temperature. The primary antibody was diluted and incubated overnight at 4°C, then washed three times with tris-buffered saline and tween 20 (TBST) at 10-minute intervals. The secondary antibody was diluted 1:1,000, incubated at room temperature for 2 hours, washed three times with TBST, and bands were identified and quantified using the Davinch-Chemi™ Imager CAS-400SM System.

Figure 8 shows the expression levels of MITF, tyrosinase, TRP-1 and TRP-2 proteins in melanoma cells (B16F10) by concentrations of a mixture of pine wood extract and kenaf aerial part extract.

Figure 8A shows the expression level of MITF at different concentrations of a mixture of pine nut wood extract and kenaf aerial part extract in melanoma cells (B16F10), and Fig. 8B shows pine nut wood extract and kenaf aerial part extract in melanoma cells (B16F10). Figure 8C shows the expression level of tyrosinase by concentration of the mixture, Figure 8C shows the expression level of TRP-1 by concentration of the mixture of pine wood extract and kenaf aerial part extract in melanoma cells (B16F10), Figure 8D is melanoma It shows the expression level of TRP-2 in cells (B16F10) at different concentrations of the mixture of pine wood extract and kenaf aerial part extract.

Referring to FIG. 8, MITF and TRP-1 increased by α-MSH in the B16F10 group treated with a mixture of pine nut wood extract and kenaf aerial part extract at concentrations of 25 μg/ml, 50 μg/ml, and 100 μg/ml. , it was confirmed that the protein expression of TRP-2 and tyrosinase was reduced, and a particularly significant result was confirmed in the protein expression of tyrosinase when compared to the control kojic acid.

<Experimental Example 9> Confirmation of whitening effect: mRNA expression inhibition measurement through reverse transcription-PCR

In order to investigate the mRNA expression inhibitory effect of the mixed extract on MITF, TRP-1, TRP-2 and tyrosinase, which are factors related to melanin synthesis, 25 μg/ml, 50 μg/ml, and 100 μg/ml of B16F10 mouse melanoma cells were tested. After processing the extract by concentration, the control was treated with α-MSH to overexpress melanin, and the normal part was set to a section not treated with α-MSH. 24 hours later, mRNA expression was measured by RT-PCR, and at this time, GAPDH, a house keeping gene with little difference in expression level under various cell conditions, was used as a positive control.

The primer sequences used in the experiment are shown in Table 1. 5 × green Go Taq Flexi buffer, MgCl2, PCR Nucleotide Mix (10 mM), primer, Go Taq DNA Polymerase, nuclease free water, and synthesized cDNA were added to the PCR tube, mixed well, and then PCR was performed. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was 96 ℃ for 10 seconds, 64 ℃ for 30 seconds, 72 ℃ 1 minute (40 cycles), and Tyrosinase was 96 ℃ 10 seconds, 64 ℃ 30 seconds, 72 ℃ 1 minute. (40 cycles). MITF, TRP-1, and TRP-2 were subjected to 96 °C for 10 seconds, 56 °C for 30 seconds, and 72 °C for 1 minute (40 cycles), respectively. After synthesis by PCR, electrophoresis was performed on a 1.5% agarose gel with 0.002% ethidium bromide at 100 V for 30 minutes, and then bands were confirmed and quantified using a UV transilluminator.

Table 1 shows the primer sequences used for RT-PCR.

Gene Primer Sequence (5’ → 3’) GAPDH Sense TGA AGG TCG GTG TGA ACG GAT TTG GC Anti-sense CAT GTA GGC CAT GAG GTC CAC CAC MITF Forward AGC GTG TAT TTT CCC CAC AG Reverse TAG CTC CTT AAT GCG GTC GT TRP-1 Forward ACT TCA CTC AAG CCA ACT GC Reverse AGC TTC CCA TCA GAT GTC GT TRP-2 Forward GCT CCA AGT GGC TGT AGA CC Reverse AAT GCA GTG GCT TGG AAA TC Tyrosinase Forward GAC GGT CAC TGC ACA CTT TG Reverse GCC ATG ACC AGG ATG AC MMP-1 Sense AGC GTG TGA CAG TAA GCT AA Anti-sense GTT TTC CTC AGA AAG AGC AGC AT MMP-2
Sense TTG CCA TCC TTC TCA AAG TTG TAG G Anti-sense CAC TGT CCA CCC CTC AGA GC MMP-3
Sense TTG TTC TTT GAT GCA GTC AGC Anti-sense GAT TTG CGC CAA AAG TGC

9 shows the expression levels of MITF, TRP-1, TRP-2, and tyrosinase mRNA in melanoma cells (B16F10) by concentrations of a mixture of pine wood extract and kenaf aerial part extract.

Figure 9A shows the expression level of MITF mRNA at different concentrations of the mixture of pine nut wood extract and kenaf aerial part extract in melanoma cells (B16F10), and Fig. 9B shows the expression level of pine nut wood extract and kenaf aerial part in melanoma cells (B16F10). Figure 9C shows the expression level of tyrosinase mRNA for each concentration of the extract mixture, and Figure 9C shows the expression level of TRP-1 mRNA for each concentration of the mixture of pine wood extract and kenaf aerial part extract in melanoma cells (B16F10), Figure 10D shows the expression level of TRP-2 mRNA in melanoma cells (B16F10) at different concentrations of the mixture of pine wood extract and kenaf aerial part extract.

Referring to FIG. 9, mRNA was overexpressed in the control part of the B16F10 group treated with a mixture of pine nut wood extract and kenaf aerial part extract at concentrations of 25 μg/ml, 50 μg/ml, and 100 μg/ml, and MITF and TRP-1 As a result of measuring the mRNA expression of TRP-2 and tyrosinase, the mRNA expression was suppressed as the concentration increased, and when compared to the control group, kojic acid, the expression of TRP-2 and tyrosinase was significantly suppressed.

As described above, although the embodiments have been described with limited drawings, those skilled in the art can apply various technical modifications and variations based on the above. For example, the described techniques may be performed in an order different from the method described, and/or components of the described system, structure, device, circuit, etc. may be combined or combined in a different form than the method described, or other components may be used. Or even if it is replaced or substituted by equivalents, appropriate results can be achieved.

Therefore, other implementations, other embodiments, and equivalents of the claims are within the scope of the following claims.

Claims (13)

Containing a mixture of a pine tree ( Pinus koraiensis ) wood extract and a kenaf ( Hibiscus cannabinus ) extract as an active ingredient,
cosmetic composition.
According to claim 1,
The weight ratio of the pine wood extract and the kenaf extract is
1: 10 to 10: 1,
cosmetic composition.
According to claim 1,
The mixture is
Of the total cosmetic composition, contained in 0.1% to 50% by weight,
cosmetic composition.
According to claim 1,
The extract is
At least one selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, acetone, ethyl acetate, chloroform, 1,3-butylene glycol, n-hexane, diethyl ether, butyl acetate and dichloromethane as an extraction solvent which is extracted by
cosmetic composition.
According to claim 4,
The extraction is
Which is carried out for 4 hours to 60 hours at a temperature of 1 ℃ to 40 ℃,
cosmetic composition.
According to claim 1,
1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid, ABTS) having radical scavenging activity or SOD (superoxide dismutase)-like activity,
cosmetic composition.
According to claim 1,
Inhibition of microphthalmia associated transcription factor (MITF) expression, inhibition of tyrosinase related protein-1 (TRP-1) expression, inhibition of tyrosinase related protein-2 (TRP-2) expression, inhibition of tyrosinase activity and elastase ) having at least one performance of activity inhibition,
cosmetic composition.
According to claim 1,
Pine tree ( Pinus koraiensis ) Which further comprises leaf ethanol extract as an active ingredient,
cosmetic composition.
According to claim 8,
Based on 100 parts by weight of the mixture, the pine nut leaf ethanol extract is contained in 10 parts by weight to 100 parts by weight,
cosmetic composition.
According to claim 1,
Further comprising, as an active ingredient, at least one vegetable extract selected from the group consisting of centella asiatica extract, propolis extract, houttuynia cordata extract, green tea leaf extract, vermiculite extract, and goji berry extract,
cosmetic composition.
According to claim 10,
With respect to 100 parts by weight of the mixture, the vegetable extract is contained in 1 part by weight to 30 parts by weight,
cosmetic composition.
According to claim 1,
The cosmetic composition,
For antioxidant, whitening, wrinkle improvement, convergence, pore contraction or inflammation inhibition,
cosmetic composition.
According to claim 1,
The cosmetics,
Any one of basic cosmetics selected from the group consisting of lotion, toner, skin, lotion, emulsion, cream, essence, serum, gel and pack, lipstick, tint, lip glow, lip balm, BB cream, foundation, powder, mascara, eye Make-up cosmetics selected from shadows and eyeliners;
A cosmetic for washing, which is any one selected from the group consisting of dead skin scrub, cleansing water, cleansing oil, cleansing cream, and foam cleansing,
A cosmetic for hair that is any one selected from the group consisting of shampoo, rinse and treatment, and
Any one cosmetic for body selected from the group consisting of body scrub, body lotion, body cream, and body wash;
To include one or more selected from the group consisting of,
cosmetic composition.

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