KR20160020384A - Cosmetic Composition Comprising Extract of cone scale of Pinus koraiensis Siebold et Zucc - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising an extract of a hull of Pinus koraiensis Siebold et Zucc. as an active component, and a skin external agent comprising the same. Also, the present invention relates to a food composition and a pharmaceutical composition comprising an extract of a hull of Pinus koraiensis Siebold et Zucc. as an active component. The extract of a hull of Pinus koraiensis Siebold etZucc. of the present invention has excellent effects on anti-oxidation, whitening, or anti-wrinkle, so the extract of the hull of Pinus koraiensis Siebold et Zucc. according to the present invention is used as a functional raw material for anti-oxidation, whitening, and anti-wrinkle.

Description

[0002] The present invention relates to a cosmetic composition comprising an extract of Pine nuts and shells as an active ingredient. [0002] The present invention relates to a cosmetic composition comprising an extract of Pinus koraiensis Siebold et Zucc

The present invention relates to a cosmetic composition comprising as an active ingredient an extract of zug and a shell and an external preparation for skin containing the same.

The present invention also relates to a food composition and a pharmaceutical composition comprising as an active ingredient an extract of zug and shell.

As well-being culture has become a trend in recent years, research on various fields such as cosmetics, foods, and pharmaceuticals has been actively carried out with the aim of improving the skin problems caused by ultraviolet rays, food, environment, etc., .

As the protective layer of the outermost layers of the body, the skin is an organ that functions to protect the internal organs from the external environment such as ultraviolet rays and harmful substances and to prevent loss of moisture and electrolytes. Because the skin is always exposed to the external environment, excessive UV exposure can cause skin burns, melanin deposition, wrinkles and photoaging, skin irritation and, in severe cases, skin cancer.

In general, melanin is made to protect the skin from external stimuli such as ultraviolet light. Over-deposition of melanin pigment on skin may result in hyperpigmentation, phototoxic response or other similar small-sized post-inflammatory reactions due to scarring or dermatitis, including unwanted freckles, senile spots, liver, spots, brown or black spots, It can cause fixed pigmented lesions.

Melanogenesis is regulated by three enzymes, tyrosinase, tyrosinase related protein-1 (TRP-1) and TRP-2. The mechanism of biosynthesis of melanin is that the starting material melanin biosynthesis is transformed into DOPA (3,4-dihydroxyphenylalanine) and DOPA quinone by tyrosinase, which is a starting material, tyrosine, as a rate-limiting enzyme, It is oxidized to the intermediate 5,6-dihydroxyindole-2-carboxylic acid to produce eumelanin. When keratinocytes in the skin are stimulated by ultraviolet rays, signaling substances such as α-MSH (α-melanocyte-sitmulating hormone) and ACTH (adrenocorricotropic hormone) are secreted into melanocytes. Melanocortin type 1 receptor (MC1R) receptors increase the level of cyclic adenosine monophosphate (cAMP) by inducing these stimuli. The increase of cAMP causes the activation of PKA (protein kinase A), followed by CREB element-binding protein). The phosphorylated CREB causes the expression of MITF in the microphthalmia associated transcription factor (MITF), a specific transcription factor of melanocytes. The expression of MITF ultimately enhances melanin biosynthesis by promoting the binding of tyrosinase, TRP-1, and TRP-2, which are major enzymes of melanin production, to the promoter.

MMPs (matrix metalloproteinase), which degrades substrates among various proteolytic enzymes, play an important role in the development of skin photo-aging as the ultraviolet rays exposed to the skin increase. Extracellular matrix proteins in the dermis play an important role in maintaining the structure and resilience of the skin, 80% to 85% of which 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, resulting in a complete collagen after the amino and carboxyl ends are cleaved. In particular, MMPs play an important role in the degradation of matrix proteins, including intestitial collagenase (MMP-1), 72 kD gelatinase (MMP-2), 92 kD gelatinase (MMP-9), neutrophil collagenase (MMP-8), and collagenase (MMP-13) In skin exposed to ultraviolet light, the expression of MMPs is increased, and the increased MMPs break down the collagen that constitutes the skin, resulting in the deficiency of the substrate protein, resulting in skin aging and wrinkles.

Pinus pinus koraiensis Siebold et Zucc) is an evergreen tree belonging to the pine tree family. It grows at an elevation of 1,000 m above sea level and grows up to 30 m in height and 1 m in diameter. The bark is gray-brown and the sculpture falls. The leaves are 3 ~ 5 at the end of the short branch, and there is a white pore line on the back, light green color, with serrate on the edge.

Traditionally, pine wood has been classified as a waste resource except for pine nuts. In particular, in the case of the cones which are the pine nut of the pine by-product, the husks (seeds) enclosed in the outer husks can be harvested only by removing the shells (outer shells), and the white nuts (endosperm) (Shells and outermost shells) are left untouched or discarded. Therefore, it is necessary to study the effective utilization methods.

Accordingly, the present inventors have found that there is a substance having an effect of whitening and wrinkle-removing in extracts of zygote and shell (zucchini and exocytosis), which are abandoned after being stripped for harvesting white nuts And have completed the present invention.

KR Application number: 10-2012-0106848

One aspect of the present invention is to provide a cosmetic composition comprising an extract of Zingiber shell and shell extract as an active ingredient and a skin external preparation containing the same.

Another aspect of the present invention is to provide a food composition and a pharmaceutical composition comprising an extract of Zygoda crustacea and a shell extract as an active ingredient.

One aspect of the present invention provides a cosmetic composition comprising as an active ingredient an extract of zug and shell.

Hereinafter, the present invention will be described in detail.

The pine nut cone of the present invention is pine (Pinus as fruit of pine koraiensis Siebold et Zucc) is an evergreen tree belonging to the pine tree family. In the present invention, the shell of the zodiacal gland refers to a piece of pine nut which is removed from the husks surrounded by the outer skin.

The pine nuts can be mature pine nuts.

The pine nuts and shell extract of the present invention may be extracted with water, C1 to C4 lower alcohols or a mixed solvent thereof. In the specific example of the present invention, extraction was performed using hot water and ethanol. The above-mentioned extract is a concept that includes all of the extracts of the above zodiacal crustaceans, the fractions thereof and the active fractions separated from the fractions.

The pine nuts and shell extracts of the present invention can be used for antioxidant, whitening, shrinkage of pores or wrinkles. In the examples of the present invention, various activities were confirmed on the extracts of zug and peel. As a result, it was confirmed that the extracts of zug and peel inhibited DPPH, PF, ABTS and TBARS and had antioxidative effect. In addition, the extract inhibited tyrosinase and inhibited melanin production by inhibiting the expression of tyrosine and MITF proteins in a B16F10 cell experiment, thereby exhibiting a whitening effect. In addition, the extracts of zygote and bark showed wrinkle-reducing effects by inhibiting esterase activity, maintaining the elastin molecular shape, activating ProCOL1A2, and inhibiting the expression of MMP-1 and MMP-9. Therefore, the pine bark extract of the present invention can be used as a functional ingredient useful for antioxidative, whitening and wrinkle-reducing effects.

The cosmetic composition of the present invention may further contain conventional auxiliary agents and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances and the like which are conventionally used in cosmetic compositions, in addition to the above-mentioned effective ingredients. For example, the cosmetic composition may further contain auxiliary ingredients such as glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, and allantoin.

The content of the zitta extract and the actual floss extract may be 0.1 to 50% by weight, more preferably 0.3 to 10% by weight, based on the total weight of the cosmetic composition, and the content thereof may be freely selected depending on the formulation, application and purpose.

For example, at this time, based on one pack (30 g) of cosmetics containing zuggu and horsetail extract,

Based on the total weight of the cosmetic composition

1) The amount of the powder of zapper and flock extract is 0.03 g at 0.1 wt%, 0.09 g at 0.3 wt%, 3 g at 10 wt%, and about 15 g at 50 wt%.

2) The amount of phenolic compounds is 0.1521 mg at 0.1% content, 0.4563 mg at 0.3% content, 15.21 mg at 10% content and 76.05 mg at 50% content

3) The weight of the raw material of the zug and the actual pieces relative to the total weight of the cosmetic composition is 9g at the content of 0.1%, 27g at the content of 0.3%, 90g at the content of 10%, and 450g at the content of 50% .

The cosmetic of the present invention may be blended with other essential ingredients, if necessary, in combination with the essential ingredients.

Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.

Examples of the oil retaining component include ester-based oil retaining, hydrocarbon-based oil retaining, silicone-based oil retaining, fluoric oil retaining, animal retention and plant retention.

Examples of ester-based fats include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearyl isostearate, Propyl isostearate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, isopropyl myristate, butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isosilyl myristate, Trimethylol propane, triisostearic acid trimethylol propane, tetra 2-ethylhexanoic acid pentaerythritol tetra (2-ethylhexanoate) , Decyl caprylate, decyl laurate, hexyl laurate, myristate decyl, myristyl myristate, myristine monoethyl stearate, stearyl stearate, decyl oleate, ricinoleic acid tri , Isostearyl stearate, isostearyl stearate, isodecyl stearate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, isopropyl stearate, isopropyl stearate, isopropyl stearate, -Hexyl stearate, stearyl ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol, Propyleneglycol propionate, propyleneglycol propionate, dicaproic acid neopentyl glycol, dioctanoic acid neopentyl glycol, tricarboxylic acid glyceryl, triunsaturated glyceryl, triisopalmitic acid glyceryl, triisostearic acid glyceryl, neopentanoic acid octyldodecyl Octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, octanoic acid octanoate, Octyldecyl lactate, octyldecyl lactate, octyldecyl lactate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisobutyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, But are not limited to, ethyl, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, But are not limited to, dioctyl sebacate, stearic acid cholesteryl, isostearic acid cholesteryl, hydroxystearic acid cholesteryl, oleic acid cholesteryl, oleic acid dihydrocholesteryl, isostearic acid pitostearyl, Stearoyl hydroxystearic acid isostearyl, 12-stearoyl stearyl hydroxystearate, 12-stearo And monohydroxystearic acid and esters such as sostearyl.

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

Examples of silicone based oils include polymethyl silicone, methylphenyl silicone, methyl cyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstarchoxysiloxane copolymer, alkyl Modified silicone oils, and amino-modified silicone oils.

Examples of the fluorine-based oil include perfluoropolyether and the like.

Examples of animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, , Corn oil, palm oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, shea butter, coltsfoot colostrum, marker daisy nut oil, mead home oil, egg oil, , Canned wax, carnauba wax, liquid lanolin, hardened castor oil, and the like.

Examples of the moisturizing agent include water-soluble low-molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and oil-soluble polymers.

Examples of the water-soluble low-molecular moisturizing agent include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B Glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactic acid salt and the like.

Examples of the lipid-soluble low-molecular moisturizing agent include cholesterol and cholesterol ester.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartic acid, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, etc. .

Examples of the oil-soluble polymer include polyvinylpyrrolidone / eicosene copolymer, polyvinylpyrrolidone / hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymer silicone.

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

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

Examples of the nonionic surfactant include self emulsifying monostearate glycerin, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbit fatty acid ester, POE (Polyoxyethylene / polyoxypropylene) copolymer, POE.POP alkyl ether, polyether-modified silicone, polyether-modified silicone, polyoxyethylene-polyoxypropylene (POE) Alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Examples of the anionic surfactant include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylallylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkylether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphate, alkylamide phosphate , Alkyloyl taurine salts, N-acyl amino acid salts, POE alkyl ether carboxylates, alkylsulfosuccinates, sodium alkylsulfoacetates, acylated hydrolyzed collagen peptide salts, and perfluoroalkyl phosphate esters .

Examples of the cationic surfactant include alkyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium bromide, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium chloride, behenyl trimethyl ammonium chloride, Benzalkonium, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salts of lanolin derivatives, and the like.

Examples of the amphoteric surfactant include carboxybetaine type, amide betaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type and amide amine type Amphoteric surfactants and the like.

Examples of the organic and inorganic pigments include inorganic pigments such as silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, Bengala, clay, bentonite, titanium mica, titanium oxide, bismuth chloride, zirconium oxide, magnesium oxide, Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, chromium oxide, chromium oxide, chromium hydroxide, But are not limited to, polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, Silk powder, cellulose, CI Pigment Yellow, CI Pigment Orange, and composite pigments of inorganic pigments and organic pigments thereof.

As the organic powder, metallic soap such as calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; Such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylidene, N-alpha-paratyylnitine, N-alpha-lauroyl arginine, Acyl basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid, alpha-aminoaurauric acid, and the like; Polyethylene, polypropylene, nylon, polymethylmethacrylate, polystyrene, divinylbenzene-styrene copolymer, ethylene tetrafluoride, and the like.

Examples of ultraviolet absorbers include paraaminobenzoic acid, ethyl parnamobenzoate, amyl paranobenzoate, octyl paranobenzoate, ethyleneglycol salicylate, phenyl salicylate, benzyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, benzyl cinnamate , Octyl methoxycinnamate, dioctyl methoxycinnamate, mono-2-ethylhexane glyceryl dipyrromethoxycinnamate, isopropyl paratumoxycinnamate, diisopropyl-diisopropyl cinnamate ester mixture, Carninoic acid, ethyl urocanoate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and salts thereof, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone , Tetrahydroxybenzophenone, 4- tert -butyl-4'-methoxydibenzoylmethane, 2,4,6-trianylino- p- (carbo-2'-ethylhexyl-1'- , 3,5-triazine, 2- (2- And the like can be mentioned hydroxy-5-methylphenyl) benzotriazole.

Examples of the disinfectant include hinokitiol, trichloroacid, trichlorohydroxydiphenyl ether, crohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zinc filitione, benzalkonium chloride, No. 301, mononitro and eicol sodium, and undecylenic acid.

Examples of the antioxidant include butylhydroxyanisole, gallic acid propyl, and eicosorbic acid.

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

Examples of the alcohol include higher alcohols such as cetyl alcohol.

In addition, any of the above components may be blended to the extent that the object and effect of the present invention are not impaired, but it is preferably 0.01-5% by weight relative to the total weight , More preferably 0.01-3% by weight.

The cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture or the like.

The ingredients contained in the cosmetic composition of the present invention may contain, in addition to the above-mentioned extracts, the components conventionally used in cosmetic compositions, such as stabilizers, solubilizers, vitamins, pigments and fragrances, And a carrier.

In addition, one aspect of the present invention provides an external preparation for skin comprising the zodiac and bark extracts. The external preparation can be used as a cosmetic for antioxidant, whitening or wrinkle improvement, and can be used as a medicine for preventing or treating pigmentation diseases.

The external preparation of the present invention can be prepared into any formulation conventionally produced by referring to the known art in the art. For example, the compositions may be formulated as solutions, suspensions, emulsions, suppositories, suppositories, pastes, gels, creams, lotions, essences, emulsions, packs, cleansers, gels, powders, Emulsion foundation, wax foundation and spray, but are not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a mask pack, a spray or a powder.

In order to use the extract as an external preparation for skin, it may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, foaming agents, Any of those commonly used in emulsifying or emulsifying agents, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, ≪ / RTI > and other ingredients of the skin. In addition, the components can be introduced in amounts commonly used in the dermatology field.

When the formulation of the present invention is a paste, cream or gel, the carrier component may include an animal oil, vegetable oil, wax, paraffin, starch, tracer, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, have.

When the formulation of the present invention is a powder or a spray, the carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and the like. Particularly in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane, dimethyl ether, and the like.

When the formulation of the present invention is a solution or an emulsion, the carrier component may include a solvent, a solubilizing agent, and an emulsifying agent. Specific examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.

When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol, propylene glycol or the like as a carrier component; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, trachant, and the like.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether Aliphatic alcohols, fatty acid glycerides, fatty acid diethanol amides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like.

In addition, one aspect of the present invention provides a food composition for antioxidant, whitening, or antiaging agent comprising an extract of Pine Nut and Peel as an active ingredient.

When the extract of the present invention is used as a food additive, it can be suitably used according to a conventional method such as adding the extract as it is, mixing it with another food or food ingredient, and the like.

The amount of the extract as the active ingredient may be suitably changed according to the purpose of use (prevention, health or therapeutic treatment), and the extract may be contained in an amount of 0.001 to 50% by weight based on the total weight of the food composition By weight, more preferably 0.1 to 30% by weight.

As a specific example, the extract of the present invention is added in an amount of 30% by weight or less, preferably 10% by weight or less, with respect to the raw material when the food or beverage is produced. However, when it is intended for health and hygiene purposes or for the purpose of controlling health, it can be added in an amount below the above range, and there is no problem in terms of safety. Therefore, the active ingredient can be used in an amount exceeding the above range have.

There is no particular limitation on the kind of the food. Examples of the food to which the extract of the present invention can be added include meat products, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, Drinks, alcoholic beverages, vitamin complexes, and the like, all of which include health foods in a conventional sense.

When the food composition of the present invention is prepared as a beverage, it may contain additional ingredients such as various flavors or natural carbohydrates such as ordinary beverages. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame, and the like. The natural carbohydrate is contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , Carbonating agents used in carbonated beverages, and the like. In addition, the compositions of the present invention may include flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of the above additives is not limited to a great extent, but may be in the range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

In addition, one aspect of the present invention provides a pharmaceutical composition for treating or preventing a pigmentation disease comprising an extract of Zygomugoschus sinensis and a skin extract as an active ingredient. These pigmentation disorders include, but are not limited to, freckles, aging spots, liver, spots, dark spots, sunspots, daylight pigmentation, cyanic melasma, hyperpigmentation after drug use, gravidic chloasma, Hyperpigmentation after inflammation due to wounds or dermatitis involved, and the like.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The present invention relates to pharmaceutical compositions comprising an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, RTI ID = 0.0 > effective < / RTI > amount. It will be apparent to those skilled in the art that the therapeutically effective dose and frequency of administration for the pharmaceutical compositions of the present invention will vary with the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. For a desired effect, the pharmaceutical composition of the present invention may be administered in an amount of 0.001 to 100 mg / kg / day, and may be administered once a day, or divided into several doses.

The pharmaceutical compositions of the present invention can be administered to a subject in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The zit composition and the peel extract of the present invention can be used for antioxidant, whitening or wrinkle improvement. In the examples of the present invention, various activities were confirmed on the extracts of zug and peel. As a result, it was confirmed that the extracts of zug and peel inhibited DPPH, PF, ABTS and TBARS and had an antioxidative effect. In addition, the extract inhibited tyrosinase and inhibited melanin production by inhibiting the expression of tyrosine and MITF proteins in B16F10 cell experiments, thereby exhibiting a whitening effect. In addition, the extracts of zygote and bark showed wrinkle-reducing effects by inhibiting esterase activity, maintaining the elastin molecular shape, activating ProCOL1A2, and inhibiting the expression of MMP-1 and MMP-9. Therefore, the pine bark extract of the present invention can be used as a functional ingredient useful for antioxidative, whitening and wrinkle-reducing effects.

FIG. 1 shows the result of measuring the content of a physiologically active component eluted by ethanol concentration using a phenolic compound as a main target.
FIG. 2 is a graph showing the oxidative free radical reactivity of water and ethanol extracts obtained from the paddy field by DPPH method.
FIG. 3 is a graph showing changes in DPPH scavenging ability according to Phenolic concentration of extracts of zug and shell. FIG.
Fig. 4 shows the results of antioxidative effects of extracts of zug and shells using ABTS radical cation decolorization.
FIG. 5 shows the results of ABTS radical cation decolorization measured according to the Phenolic concentration of the extracts of Zug and Pseudomonas sp.
FIG. 6 shows the result of measurement of the antioxidant protection factor (PF) of the extract of Zug and the actual flock.
FIG. 7 shows the result of measuring the antioxidant protection factor (PF) according to the Phenolic concentration of the extracts of zug and obsidian.
FIG. 8 is a graph showing the results of TBARS measurement of the extracts of zug and obsidian.
FIG. 9 is a graph showing the results of TBARS measurement according to the Phenolic concentration of the extracts of zug and obsidian.
FIG. 10 is a graph showing the convergence effect of the extract of Zug and the actual grain fractions. FIG.
FIG. 11 is a graph showing the tyrosinase inhibitory activity of the extract of zug and obsidian.
FIG. 12 is a graph showing the survival rate of melanoma cell (B16F10) according to the treatment of the extract of Zygometaceae and arthropods. A: water extract, B: ethanol extract.
FIG. 13 is a graph showing tyrosinase inhibitory activity in melanoma cell (B16F10) according to treatment of zug and obsolete extracts. A: water extract, B: ethanol extract.
FIG. 14 is a graph showing inhibition of melanin biosynthesis in melanoma cell (B16F10) according to the treatment of extracts of zug and obsidian. A: water extract, B: ethanol extract.
FIG. 15 is a graph showing the inhibitory effect of tyrosinase on melanin synthesis in extracts of Zygote and seedlings. 1, 5, and 10 μg / mL of the extracts were treated for 24 hours. Tyrosinase was detected by Western blot using antibody. Diagrams were derived from normalization based on β-actin. Each value was expressed as mean ± SD of three independent experiments.
FIG. 16 is a graph showing the effect of suppressing the expression of microphthalmia-associated transcription factor (MITF) involved in the synthesis of melanin in the extracts of Zug and Actival.
FIG. 17 is a view showing the elastase inhibitory activity of the extracts of zug and obsidian.
FIG. 18 is a graph showing collagenase inhibitory activity of extracts of Zug and Actival.
19 is a graph showing the results of MTT measurement of CCD986sk fibroblast cells according to treatment of zug and obsolete extracts. A: water extract, B: ethanol extract.
FIG. 20 shows the result of confirming the pro-collagen biosynthesis rate according to the treatment of the extracts of zug and obsidian. A: water extract, B: ethanol extract.
FIG. 21 is a graph showing the inhibitory effect of procollagenase protein on the treatment of zug and obsolete extracts.
FIG. 22 shows the results of confirming the amount of wrinkle-related protein (MMP-1) and mRNA expression of fibroblast cells (CCD-986sk) treated with the extracts of zug and obsidian. A: water extract, B: ethanol extract.
23 and 24 are graphs showing the inhibitory effect of intestitial collagenase (MMP-1) and 92 kD gelatinase (MMP-9) on protein expression in fibroblast (CCD-986sk)
Fig. 25 shows the results of TIMP-1 activity according to the treatment of zug and obelisk extract. A: water extract, B: ethanol extract.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example 1. Preparation of materials

The materials used in this study were naturally grown in Pinus koraiensis in Gapyeong, Gyeonggi Province, and the matured pine nuts were collected from the pine nuts and separated from the shells and shells surrounding the outer surface of the pine nuts. . The seeds were removed from the husk as a fruit (white pine nut) by removing the outer husks. The separated seeds and outer slices were washed, washed, and then pulverized to about 40 mesh.

Example 2. Pine nut  skin( Cone Actual piece ) Preparation of extract

In the case of hot water extract of zug and shell, 100 g of the sample is added with 10 times of distilled water and refluxed for 3 hours at 85 ° C. The supernatant is separated from the precipitate and extracted three times. In the case of the ethanol extract, And the mixture was immersed and extracted at room temperature for 24 hours, followed by repeating the above three times by separating the supernatant and the precipitate. Each extract was centrifuged at 12,000 rpm for 10 minutes. 1 filter paper, and each extract was concentrated and stored in the freezer after freeze drying, and used as a sample of this experiment.

* Example 3. Total polyphenol content measurement

Polyphenol compounds were measured by the Folin-Denis method, the sample 1 mL added to 95% ethanol 1 mL) and distilled water (5 mL to give mixed well into a 1N Folin-ciocalteu reagent 0.5 mL, and then allowed to stand for 5 minutes, Na ₂ After adding 1 mL of CO ₃ , the absorbance was measured at 725 nm within 1 hour and the amount was converted from the standard curve using gallic acid.

Example 4: pine nuts, conifers bark (coniferous Silpyeon and oejongpi) electron donating ability (EDA extracts of: electron donating ability) measurement

The electron donating ability (EDA) is measured by modifying the Blois method. Add 1 ml of 0.2 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) to 2 ml of each sample solution, stir and incubate for 30 minutes, and measure the absorbance at 517 nm. The electron donating ability is represented by the absorbance reduction rate of the sample solution and the no-added solution.

Example  5. ABTS  radical cation decolorization ( ABTS ) Measure

ABTS radical cation decolorization was measured by Pellegrin et al. That is, 7 mM ABTS and 140 mM K ₂ S ₂ O ₈ were mixed with 5 μL of 88 μL, left in the dark for 14-16 hours, and then mixed with absolute ethanol at a ratio of 1:88. The absorbance value of the control was 734 nm 0.7 ± 0.002 was used. As a positive control, 50 μL of BHT and 1 mL of ABTS solution were mixed for 30 seconds, incubated for 2.5 minutes, and the absorbance was measured at 734 nm. The inhibition rate (%) was calculated by the following equation.

Example  6. Antioxidant protection factor ( PF ) Measure

PF was measured by the method of Andarwulan and Shetty. 10 mL of β-carotene was dissolved in 50 mL of chloroform, and 1 mL of the solution was added to the evaporator. The chloroform was distilled in a water bath at 40 ° C., and 20 μL of linoleic acid, 184 μL of Tween 40 and 50 mL of H 2 O 2 were added to make an emulsion , 5 mL of the emulsion was mixed with 100 μL of the sample solution, vortexed well, reacted at 50 ° C. for 30 minutes, and then the absorbance was measured at 470 nm.

Example  7. TBARs  ( Thiobarbituric  acid reaction substance

TBARS was measured according to the method of Blurge and Aust. The emulsion was prepared with 1% linoleic acid and 1% Tween 40, mixed with 0.8 mL of emulsion and 0.2 mL of sample, and reacted in a water bath at 50 ° C for 10 hours. After the reaction, add 2 mL of TBA regent [3 mL of 2% BHT / EtOH mixture] to 100 mL stock solution (15% trichloroacetic acid (TCA), 0.375% thiobarbituric acid, 0.25 M HCl mixture) After boiling and cooling for 10 minutes, the mixture was centrifuged for 15 minutes at 1000 rpm (vision, VS-5500N), and incubated at room temperature for 10 minutes. Absorbance was measured at 532 ㎚ for TBARS. The product was labeled with μM of 1,1,3,3-tetraethoxy propane (TEP) produced.

Example 8. Measurement of convergence effect

Astringent activity was measured by Lee et al. Using hemoglobin similar to skin protein, each sample solution and hemoglobin solution were mixed in a ratio of 1: 1 into a centrifuge tube, mixed with shaking, centrifuged at 1,500 rpm for 3 minutes, and absorbance was measured at 576 nm. The astringent activity measurement was expressed by the absorbance reduction ratio of the sample solution addition group and the no addition group.

Example 9. Measurement of whitening effect

(One) Tyrosinase  Measurement of inhibitory activity

Tyrosinase inhibitory activity was measured by Yagi et al. To the reaction mixture, 0.2 mL of mushroom tyrosinase (110 U / mL) was added to a mixture of 0.2 mL of the substrate solution in which 10 mM L-DOPA was dissolved in 0.5 mL of 0.175 M sodium phosphate buffer (pH 6.8) and 0.1 mL of the sample solution and the mixture was reacted at 25 ° C. for 2 minutes And the DOPA chrome produced in the reaction solution was measured at 475 nm. The tyrosinase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample.

(2) Pine nut  skin( Cone Actual piece ) Cell line whitening efficacy of extract

1) Cell viability measurement

(A) Cell culture

Cultures of each cell used for this experiment is 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (100U / mL) a dulbeco's modified eagle's medium (DMEM ) was used as the medium, 37 ℃, 5% CO ₂ was added to incubator and subcultured.

(B) Measurement of cell viability by MTT assay

Cell viability was measured by the method of Carmichael et al. Cells were seeded in 96-well plates at a concentration of 0.6 to 8 × 10 ³ cells / well (0.18 ml), and the concentrations of each cell line (melanoma (B16F10), fibroblast (CCD-986sk) and macrophage mL, and incubated at 37 ° C in a 5% CO ₂ incubator for 24 hours. In the control group, the same amount of distilled water as that of the sample was added and the cells were cultured under the same conditions. After adding 0.02 mL of MTT solution (5 mg / mL) for 4 hours, the culture solution was removed and 0.15 mL of DMSO: ethanol (1: 1) was added to each well. Absorbance at 550 nm.

2) Measurement of cellular tyrosinase inhibitory activity

Melanoma (B16F10) cells were cultured in each culture dish, and the medium was removed and washed with PBS. 100 μl of lysis buffer (67 mM sodium phosphate buffer, 1% triton X-100, 0.1 mM phenylmethyl sulfonyfluoride) was added to each dish, and the cells were disrupted by ice, and cells were collected and subjected to ultra sonication. The supernatant obtained by centrifugation at 13,200 rpm for 20 minutes was used as an enzyme solution. 120 uL of 8.0 mM L-DOPA dissolved in 67 mM phosphate buffer (pH 6.8) and 40 uL of the sample solution were added to a 96-well plate and then 40 uL of the enzyme solution dissolved in 67 mM phosphate buffer (pH 6.8) After minute incubation, the amount of DOPA chrome produced was measured at 490 nm.

3) Inhibition rate of melanin biosynthesis in melanoma cell (B16F10)

Measurement of inhibition of melanin biosynthesis from skin melanoma cells was performed according to the method of Hosoi et al. B16F10 cells cultured in DMEM medium were divided into 1 × 10 ⁶ cells / dish in a 100 mm culture dish. After culturing for 24 hours, 2 mL of each sample was prepared and the cells were washed with phosphate buffer (pH 7.4) 48 hours later. After the cells were desorbed with 0.25 M trypsin-EDTA solution, the harvested cells were treated with 1 mL of 5% TCA per 1 × 10 ⁶ cells, centrifuged twice at 2,500 rpm, and the separated melanin was washed with phosphate buffer Then, 1 mL of ether: ethanol (1: 3) was added, and the mixture was centrifuged twice. 1N NaOH was added to the dried melanin, reacted at 80 ° C for 1 hour, and the absorbance was measured at 405 nm by a spectrophotometer. The inhibition of melanin biosynthesis was expressed by the absorbance reduction ratio of the sample solution and the non - added sample.

4) Expression of intracellular pigmentation-related proteins by Western blot

The amount of MITF and tyrosinase protein expression was measured by the following method. In other words, B16F10 was cultured in a culture dish, and the sample was treated. After 48 hours, the cells were washed with PBS, centrifuged, and cell lysate was prepared using RIPA buffer. The amount of protein contained in the cell lysate was measured by the Bradford method. 15 μg of protein was separated by SDS-PAGE containing 8% polyacrylamide and electroblotted with PVDF membrane at 120 V for 1 hour. The PVDF membrane is then left in 5% skim milk for 1 hour and then overnight at 4 ° C in primary antibody. After washing three times with TBST, the secondary antibody was reacted at room temperature for 1 hour, washed three times with TBST and exposed to X-ray film by reaction with ECL kit (Amersham-Pharmacia). The developed bands on the X-ray film were quantified using a LAS 4000 image analyzer.

Example 10 Effect of improving wrinkles

(One) Elastase  Measurement of inhibitory activity

Elastase inhibitory activity was measured by Cannell et al. N-succinyl- (L-Ala) ₃ - p- nitroanilide was used as a substrate and the amount of p- nitroanilide produced from the substrate at 37 ° C for 20 minutes was measured at 405 nm. 0.5 mL of each test solution was added to 0.5 mL of each test solution. 0.5 mL of porcine pancreas elastase (2.5 U / mL) dissolved in 50 mM tris-HCl buffer (pH 8.6) was added to the test tube. 50 mM tris- N-succinyl- (L-Ala) ₃ - p- nitroanilide (0.5 mg / mL) dissolved in HCl buffer (pH 8.6) was added and the reaction was allowed to proceed for 20 minutes. Elastase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample.

(2) Collagenase  Measurement of inhibitory activity

Collagenase inhibitory activity was measured by Wunsch et al. That is the reaction substrates dissolved phrase 0.1M tris-HCl buffer subject to 4mM CaCl ₂ in (pH 7.5) was added, 4-phenylazobenzyloxy carbonyl-Pro- Leu-Gly-Pro-D-Arg (0.3mg / mL) solution, and 0.25mL 0.1 mL of collagenase (0.2 mg / mL) was added to the mixture of 0.1 mL of the sample solution. The mixture was allowed to stand at room temperature for 20 minutes, and then the reaction was stopped by adding 0.5 mL of 6% citric acid. Absorbance was measured. Collagenase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample.

(3) Pine nut  skin( Cone Actual piece ) Extract's wrinkle-reducing effect on cell line

1) Cell viability measurement

A) Cell culture

Cultures of each cell used for this experiment is 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (100U / ml) a dulbeco's modified eagle's medium (DMEM ) was used as the medium, 37 ℃, 5% CO ₂ was added to incubator and subcultured.

B) Measurement of cell viability by MTT assay

Cell viability was measured by the method of Carmichael et al. Cells were seeded in 96-well plates at a concentration of 0.6 to 8 × 10 ³ cells / well (0.18 ml), and the concentrations of each cell line (melanoma (B16F10), fibroblast (CCD-986sk) and macrophage mL, and incubated at 37 ° C in a 5% CO ₂ incubator for 24 hours. In the control group, the same amount of distilled water as that of the sample was added and the cells were cultured under the same conditions. After adding 0.02 mL of MTT solution (5 mg / mL) for 4 hours, the culture solution was removed and 0.15 mL of DMSO: ethanol (1: 1) was added to each well. Absorbance at 550 nm.

(2) Skin regeneration effect of extracts of zug and skin

A) Measurement of wrinkle improvement effect using ELISA kit

i) Procollagen type I biosynthetic kit measurement

Cells were inoculated into 12 well plates at a concentration of 5x10 ⁴ cells / well, and samples were added to each well and cultured in a CO ₂ incubator for 48 hours. The cultures of the cells thus obtained were collected and used for the experiments. The degree of collagen biosynthesis in the cell culture medium was measured by using procollagen type-Ⅰ C peptide (PIP) EIA kit.

ii) Measurement of MMP-1 & TIMP-1 active kit

Cells were inoculated into 12 well plates at a concentration of 5x10 ⁴ cells / well, and samples were added to each well and cultured in a CO ₂ incubator for 48 hours. At this time, TNF-α was added at a concentration of 10 ng / mL to increase the activity of MMP-1 and TIMP-1. Cell cultures were collected and MMP-1 was measured using Amersham matrix metallproteinase-1, Human, Biotrak ELISA system (GE Healthcare).

B) Expression of intracellular wrinkle-related protein by Western blot

The amounts of MMP-1, MMP-2, and ProCOL1A2 proteins were measured using Western blot. After culturing in a culture dish using CCD-986sk, the cells were sampled and washed for 48 hours with PBS. Cells were collected by centrifugation and cell lysate was prepared using RIPA buffer. The amount of protein contained in the cell lysate was measured by the Bradford method. 20 μg of protein was separated by SDS-PAGE containing 10% polyacrylamide and electroblotted with PVDF membrane at 120 V for 1 hour. The PVDF membrane is then left in 5% skim milk for 1 hour and then overnight at 4 ° C in primary antibody. After washing three times with TBST, secondary antibody was reacted at room temperature for 1 hour, washed three times with TBST, and exposed to X-ray film by reaction with ECL kit (Amersham-Pharmacia). The bands developed on the X-ray film were quantified using a LAS4000 image analyzer.

Example 11 waste pine nut (Pinus koraiensis Siebold et Zucc) coniferous bark (coniferous Phenol extraction of material from the chamber side)

In order to harvest the fruit from the pine nuts harvested mainly from the pine trees cultivated in the Gyeonggi province, the effective ingredients of the samples can be extracted more efficiently by using the spent waste nuts and shells Water and ethanol were selected as non - harmful solvents for human body. The physiologically active components eluted by ethanol concentration were determined by using phenolic compound as a main target. The results are shown in Fig.

As shown in FIG. 1, the elution of the phenolic compound was the highest when 60% ethanol was used as a solvent, and the dissolution rate was similar in the concentration range of 40 to 60%. Table 1 shows the elution amounts of phenolic compounds under optimum conditions. As shown in Table 1, the content of water extract was 8.84 ± 0.51 mg / mL, and that of 60% ethanol extract was 5.07 ± 0.31 mg / mL.

Example  12. Pine nut Actual piece  Identification of antioxidant activity of extract

(One) Pine nut Actual piece  Extract DPPH Scatters  Measure

The DPPH method, which is an analysis of reducing materials using the oxidative free radical reaction of zapper, seaweed and ethanol extracts, was conducted. The results are shown in FIG.

As shown in FIG. 2, the maximum DPPH elimination ability was obtained in the 60% ethanol extract of the phenolic compound, and the ethanol extract and the water extract were superior in both effects.

The results of examining the DPPH scavenging ability according to the phenolic concentration are shown in FIG. As shown in FIG. 3, DPPH elimination was also increased in a concentration-dependent manner as the amount of phenolic added was increased. Also, at a low concentration of 50 μg / mL, water extracts showed high inhibition rate of 60.95% and ethanol extracts showed 88.46% The antioxidant activity of the extracts of Zingiber fragrant and Pseudomonas sp. In addition, ethanol extracts showed more excellent antioxidant ability than BHA, which is a control antioxidant, and it could be used as a material for anti - aging cosmetics.

(2) Pine nut Actual piece  Extract ABTS  radical cation decolorization  Measure

ABTS radical cation decolorization was measured for the relative antioxidant activity of the extracts. ABTS radical cation decolorization can measure both hydrogen-donating antioxidants and chain breacking antioxidants and is applicable to both aqueous and organic phases. This is based on the principle that ABTS ⁺ free radicals generated by the reaction with potassium persulfate are removed by antioxidants in the extract so that relative comparison of the extracts is possible by the use of standard substances, and the radical specific cyan color is discolored. The results are shown in Fig.

As shown in FIG. 4, it was confirmed that the hydrothermal extract showed an excellent inhibitory power of 99.21% and the ethanol extract showed the maximum inhibitory effect of 50% ethanol extract to 56.71%.

The antioxidant activity of the extracts of Zug and Pseudomonas sp. Were examined according to the contents of phenolic compounds. The results are shown in FIG.

As shown in FIG. 5, the ABTS inhibitory potency was increased as the concentration of the added phenolics was increased, and the antioxidant activity was higher than that of the control, BHA, in the range of 50-200 μg / mL phenolics , And water extracts showed high antioxidant activity (77.08%) and ethanol extract (64.96%) at low concentration of 50 μg / mL phenolics.

(3) Pine nut Actual piece  The antioxidant protection factor of the extract ( PF ) Measure

β-carotene was added to the linoleic acid emulsion to measure the antioxidant activity of the extracts of zug and obsidian for lipid-soluble substances. The results are shown in FIG.

As shown in FIG. 6, the ethanol extract was higher than the 2.54 PF of the water extract by 2.76 PF in the 60% ethanol extract.

The results of examining the antioxidant activity according to the contents of the phenolic compounds added in the extracts of Zug and Pyrite are shown in FIG.

As shown in FIG. 7, it was confirmed that as the concentration of the added phenolics increases, the PF increases in a concentration-dependent manner. The water extracts and alcohol extracts showed 1.5 ~ 2.66 PF and 1.77 ~ 3.75 PF, respectively, in the total concentration of 50 ~ 200 μg / mL phenolics. The PF was higher than 1.01 ~ 1.16 PF of the control BHA, Was very high.

(4) Pine nut  Extract TBARS  Measure

In order to measure the antioxidant activity against lipid-soluble substances, the TBARS of the extract of Zug and the actual piece was measured and the results are shown in FIG.

As shown in FIG. 8, inhibition of 66.07% and 93.10% of water extract and 60% ethanol extract was confirmed, respectively.

The results of the comparison of the antioxidant power according to the contents of the phenolic compounds added in the extracts of Zugu and Pseudomonas are shown in Fig.

As shown in FIG. 9, the higher the concentration of the added phenolics, the higher the concentration-dependent PF, and the higher antioxidant power was 90% even at a low concentration of 50 μg / mL phenolics. The water extracts were slightly lower in the whole concentration range than the control BHA, but the ethanol extracts showed better antioxidative activities than the control BHA in the concentration range of 50-200 μg / mL phenolics.

Example 13. Pine nut Actual piece  Convergence effect of extract

The principle of convergence is the phenomenon that the skin is contracted by binding with the polymer flavonoids of the skin protein to form crosslinks. Convergence means basically wrinkled or funny. Therefore, astringent agent has a function of contraction of skin and mucosal blood vessels, and has an effect of inhibiting secretion of mucus by blocking cell gap and lymphatic gap. In addition, since astringent agent has a property of binding to protein, it can generally judge the degree of astringent effect according to the degree of binding of hemoglobin protein to the extract. Such convergent action has the effect of forming an insoluble film on the surface of skin and mucous membranes by external use or contents and consequently protecting the structure or densifying the tissue to reduce the permeability of the cell membrane.

Fig. 10 shows the results of the convergence of the extracts prepared by controlling the concentration of the extracts of zug and obsidian with 50 to 200 μg / mL phenolics.

As shown in FIG. 10, the convergence effect was less than 20% in the extract of Zuggu and Pseudomonas aeruginosa. However, when ethanol extract was added at a concentration of 200 μg / mL phenolics, 62.4% of the tannic acid was slightly And 37.1%, respectively, and it could be used as a cosmetic material for reducing pores.

Example 14. Pine nut Actual piece  The result of whitening effect measurement of extract

(1) Mushroom-derived tyrosinase  Confirm inhibitory activity

Melanin, a key determinant of skin tone, is biosynthesized in melanosomes in pigmented cells called melanocytes in the epidermal basal layer. The starting material for the synthesis of melanin is tyrosine, an amino acid. Tyrosine is oxidized by tyrosinase in melanocytes to L-3,4-dihydroxyl phenylalanine (DOPA) and DOPA quinone. It is known that DOPAquinone then becomes DOPA chrome, 5,6-dihydroxyindole, and indole-5,6-quinone, and then melanin by polymerization to indole-5,6-quinone. Tyrosinase plays an important role in the formation of melanin in the skin, tyrosine hydroxylase which oxidizes tyrosine in the melanosome to form DOPA, which acts as a DOPA oxidase which oxidizes DOPA to form DOPA quinone and is an important enzyme for synthesizing melanin polymer .

The skin color of a person is determined by the pigment content such as melanin or carotene, the distribution of blood vessels, the thickness of the stratum corneum, etc. This is because melanin is the most important factor for determining skin color, Excessive synthesis or anomalies in melanin production may lead to melasma, freckle and hyper-pigmentation. Therefore, by inhibiting tyrosinase, which degrades tyrosine, a substrate of melanin synthesis, it inhibits the synthesis of melanin, and thus the whitening effect of the skin can be seen. Recently, natural and synthetic whitening agents such as vitamin C, kojic acid, arbutin, and hydroquinone have been developed and applied to whitening cosmetics for the tyrosinase inhibition effect, which is closely related to antioxidant activity.

In the present invention, tyrosinase inhibition test was carried out by adjusting the concentrations of extracts of zug and obsidian to 50, 100, 150 and 200 μg / mL. The results are shown in Fig.

As shown in FIG. 11, the inhibition effect of pine nut extract was 13.8 ~ 40% and the extract of 60% ethanol showed a high tyrosinase inhibitory effect of 18.4 ~ 41.1%. The higher the concentration of phenolic compounds, the higher the whitening effect, the lower the concentration of 100 μg / mL, the better or similar level of whitening effect than the control kojic acid, and 150 μg / mL At higher concentrations than kojic acid, tyrosinase inhibitory effect was slightly lower than that of kojic acid.

(2) Confirmation of survival rate of melanoma cell (B16F10)

MTT detection method is widely used as a cytotoxicity and cell proliferation detection method because many samples can be easily read using an ELISA reader using a 96-well plate, and the dehydrogenase activity of mitochondria in cell metabolism To reduce the yellow water-soluble MTT tetrazolium to purple colored water-insoluble MTT formazan.

The survival rate of melanoma cells by the extracts of Zug and Pseudomonas were confirmed by MTT assay at the concentrations of 1, 5, 10 and 25 μg / mL in B16F10 cells. The results are shown in Fig.

As shown in FIG. 12, cytotoxicities of 20% and 45% were obtained at a concentration of 25 μg / mL of water and ethanol extract of zucchini, respectively, and cell viability of 95% or more was observed at 1, 5 and 10 μg / Respectively.

(3) Melanoma cell (B16F10) tyrosinase  Confirm inhibitory activity

Tyrosinase is an enzyme that is most important for melanin production in tyrosine when skin is exposed to ultraviolet light. By inhibiting tyrosinase activity, it can inhibit melanin production, thereby preventing the occurrence of spots, freckles, and erythema .

tyrosinase is an enzyme that binds to Cu ^{2 +} and is a polyphenol oxidase widely distributed in plants, microorganisms, and humans. It is an enzyme that represents a major regulatory step in melanin synthesis, such as limiting the rate of melanin synthesis.

The amount of L-DOPA produced by adding zapper and roots extract to the concentrations of 1, 5, and 10 μg / mL was measured. The results are shown in FIG.

As shown in FIG. 13, inhibition rates of 20% and 30% were obtained at 10 μg / mL concentration of the extracts of Zug, Opuntia, and Alcohol, respectively, as in FIG.

(4) Confirmation of inhibition of melanin biosynthesis in melanoma cell (B16F10)

Melanin, which is the most important factor in determining the skin color of a person, not only suppresses skin photoaging and sunburn keratosis but also causes partial hyperpigmentation of spots, freckles, and black spots. Melanin is transmitted to the surrounding keratinocyte through the dendrites of the melanocyte of the epidermal basal layer and migrates to the stratum corneum of the skin. Skin blackening is due to the fact that melanocytes present in the skin increase the production of melanin in response to external conditions such as UV exposure. In order to confirm the whitening effect of raw materials and substances, the inhibition of enzyme activity of tyrosinase, which is a melanin-producing enzyme, has been mainly used. However, in recent years, in addition to suppression of enzyme activity, whitening agents have been shown to increase tyrosinase and related enzyme expression in melanocyte There have been many reports on the mechanism studies that disturb

In order to measure the content of melanin in B16F10 cells, the results of treatment with the extracts of pine needle, spruce, and alcohol extracts at the concentrations of 1, 5, and 10 μg / mL are shown in FIG.

As shown in Fig. 14, at the concentration of 10 μg / mL, the amount of melanin was lowered to 40% of that of the control group, and melanin synthesis was inhibited to the level of the normal group. In addition, it inhibited the biosynthesis of melanin by treatment with a low concentration of 1 and 5 μg / mL, so that it could be industrialized as a functional cosmetic material using a whitening effect.

(5) pine nuts, conifers Tyrosinase expression inhibitory effect involved in melanin synthesis in extracts silpyeon

To investigate the effect of tyrosinase, an enzyme involved in the synthesis of melanin, on the synthesis of melanin, the tyrosinase protein expression was determined by western blotting after 48 h of treatment with B10F10 mouse melanoma cells at a concentration of 1 to 10 μg / mL. Β-actin, a house keeping gene, was used as a positive control with little difference in the expression level of the cells under various conditions. The results are shown in Fig.

Fig. As shown in Fig. 15, the protein expression of each transcription factor was increased in the untreated B16F10 group, but the expression of tyrosinase protein was decreased in the B16F10 group treated with zapper and scrapie extract than in the untreated group, / mL inhibited the expression of water and alcohol extracts by over 60%. Therefore, it was confirmed that the extracts of Zugu and Pseudomonas spp. Inhibited the expression of mushroom tyrosinase in a dose - dependent manner.

(6) microphthalmiaassociatedtranscription involved in Melanin Synthesis factor (MITF) expression inhibitory effect

A wide variety of signaling molecules are involved in the melanin biosynthetic signaling system. Melanin is synthesized through several intracellular signal transduction mechanisms. Among them, cAMP (cyclic adenosine monophosphate) / PKA (protein kinase A) pathway is the main pathway for melanin synthesis. And activates PKA, a downstream signaling substance, and increases the expression of MITF (microphthalmia -associated transcription factor) via CREB (cAMP response element binding protein). MITF promotes the transcription of tyrosinase as an important transcription factor in melanin synthesis.

To investigate the effect of MITF on the expression of MITF for the effect of melanin synthesis on the whitening effect of extracts of Zug and Sophorae, B16F10 mouse melanoma cells were treated with 1 ~ 10 μg / blotting. At this time, β-actin, a house keeping gene with little difference in expression level, was used as a positive control even in various cell conditions. The results are shown in Fig.

As shown in Fig. 16, the protein expression of each transcription factor was increased in the B16F10 group not treated with the extract, but the expression of MITF protein was decreased in a concentration-dependent manner in the B16F10 treated group At a concentration of 10 μg / mL, the water extract showed about 30% inhibition and the alcohol extract showed about 60% inhibitory effect. Thus, it was confirmed that the alcohol extract was more effective in inhibiting MITF expression as a transcription factor than water extract. Therefore, it was confirmed that the extracts of Zugu and Pseudomonas spp. Were effective in inhibiting melanogenesis, as the result of concentration - dependent reduction of MITF expression. It was confirmed that the protein expression of tyrosinase was decreased by suppression of MITF expression in a study on the whitening effect of zug and camellia extract.

Example  15. Pine nut Actual piece  Wrinkle improvement effect measurement result of extract

(One) Elastase  Confirm inhibitory activity

Skin is a representative form of aging, such as wrinkles increase, elasticity decrease, pigmentation, etc. The most typical phenomenon of aging is wrinkles. The elastase present in the neutrophil granules of the human body is an enzyme that decomposes elastin, which is the main substrate protein for maintaining skin elasticity in the dermis. As the elastase acts, it decomposes elastin in the skin tissue, thereby losing elasticity and freshness of the skin. It causes roughness and promotes aging of the skin. Thus, inhibition of elastase activity may delay skin aging.

Elastase is a nonspecific hydrolytic enzyme capable of degrading collagen, another important substrate protein. Elastase inhibitors improve skin wrinkles. Ursolic acid is currently used as an elastase inhibitor. In addition, Cannell et al. Is one of the leucocyte granulase enzymes that degrade elastin in the body. In the abnormal tissues, the activity of the enzymes is extremely high, which is a direct cause of tissue destruction, resulting in wrinkles and loss of elasticity. Therefore, the effect of improving wrinkles can be expected by measuring the elastase inhibitory activity. Elastase activity is measured by measuring the change in color caused by the decomposition of nitroanilide (NA) using the substrate N-succinyl- (Ala) -3-p-nitroanilide using absorbance. It has the disadvantage of measuring the activity using the most widely used method or synthesized substrate instead of bioelastic.

Elastase inhibitors include ursolic acid is used as an elastase inhibitor. ursolic acid is difficult to formulate because it has insoluble properties in a solvent such as water or oil, so that it is generally difficult to use.

The elastase inhibitory activity was measured by adjusting the concentration of the extract of Zug and the actual flake to a concentration of 50 to 200 μg / mL of phenolics. The results are shown in FIG.

As shown in FIG. 17, it was confirmed that the inhibitory activity against elastase was increased in a concentration-dependent manner as the concentration of 50 μg / mL phenolis was increased from 200 μg / mL to that of the extract of zapugi and scrapie, and the addition of 200 μg / mL phenolics The inhibitory power of about 20% was confirmed.

(2) Collagenase  Inhibitory activity

Collagen, a major component of the extracellular matrix, is a major substrate protein produced by the fibroblasts of the skin. It is also an important protein that accounts for about 30% of the total biomass protein weight, and has a solid triple helix structure. Collagen forms most of the organic matter in the skin, tendons, bones and teeth, especially in the bones and dermis of the skin. The main functions of collagen are known to be mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, induction of cell division and differentiation. Such collagen is reduced by aging and photoaging by ultraviolet irradiation, which is known to be closely related to wrinkling of the skin. In addition, collagen is not affected by proteases such as trypsin but is reported to be degraded by collagenase.

The collagenase inhibitory activity was measured by controlling the concentration of the extract of Zug and the actual flock with 50 to 200 μg / mL phenolics. The results are shown in FIG.

As shown in FIG. 18, inhibition effect of collagenase was 5.02 ~ 29.83% in the extract of Zuggu and Pseudomonas sp., And inhibition effect of 5.17 ~ 46.1% in the ethanol extract. The inhibitory effect of alcohol extract on collagenase Was better. These results suggest that the extracts of garlic and tangle flour inhibit the activity of collagenase in the skin, thus preventing decomposition of collagen.

(3) Confirmation of survival rate of fibroblast cell (CCD-986sk)

1) MTT measurement results on CCD986sk fibroblst cells

MTT detection method is widely used as a cytotoxicity and cell proliferation detection method because many samples can be easily read using an ELISA reader using a 96-well plate, and the dehydrogenase activity of mitochondria in cell metabolism To reduce the yellow water-soluble MTT tetrazolium to purple colored water-insoluble MTT formazan.

In order to measure the cell survival rate of the extracts of Zug and Pseudomonas sp., CCD-986sk cells were treated at the concentrations of 1, 5, 10 and 25 μg / mL, and the results are shown in FIG.

As shown in FIG. 19, both the water extract and the alcohol extract showed cytotoxicity of about 40% at the concentration of 25 μg / mL, but the survival rate was high at the concentrations of 1, 5, and 10 μg / mL.

(4) Measurement of procollagen type I biosynthesis kit in fibroblast (CCD-986sk) using ELISA kit

Most of the collagen is present in the dermis of the skin and accounts for about 70 to 80% of the dry weight of the skin. It plays a role of supporting the skin occupying most of the extracellular matrix. However, degradation of collagen is accelerated by extrinsic factors such as decrease of collagen biosynthesis due to internal factors such as decrease of cell activity due to natural aging, increase of stress due to various harmful environments, and increase of active oxygen species by sunlight And the skin substrate is destroyed to form wrinkles. Collagen also plays an important role in healing wounds and can quickly restore scars without scarring, promoting the synthesis of collagen in damaged epithelium. The most prominent examples of collagen synthesis promoting substances that have been found so far include asiaticoside, asiatic acid, and madecasic acid, which are the major components of centella asiatica.

The synthesis of pro-collagen was measured at 1, 5, and 10 μg / mL concentrations of chitosan, disintegrant, and alcohol extracts in CCD-986sk cells. The results are shown in FIG.

As shown in FIG. 20, the extract showed pro-collagen biosynthesis in a concentration dependent manner at a concentration of 1 to 10 μg / mL. Especially, at a concentration of 10 μg / mL, the biosynthesis rate was about 70%, and the same concentration of epigallocatechin -3-O-gallate (EGCG) at a concentration of 72%.

(5) Inhibition of procollagenase protein expression in fibroblast (CCD-986sk) by Western blotting analysis

The wrinkles of the skin are formed by collagen biosynthesis being reduced, collagen degradation accelerated by various extrinsic factors such as harmful environment, and destruction of the skin substrate. The biosynthesis of collagen plays an important role in wound healing and promotes the synthesis of collagen in the damaged epithelium, thus promptly restoring the wound.

Therefore, we investigated the effect of chitosan extracts on the protein expression of procollagenase, an enzyme involved in the biosynthesis of collagen, by Western blotting the expression of procollagenase protein in CCD-986sk cells. GAPDH, a house keeping gene, was used as a positive control with little difference in the expression level even in various cell conditions. The results are shown in Fig.

As shown in Fig. 21, in the control group irradiated with UV-B, it was confirmed that the degree of procollagenase expression was remarkably decreased compared to the normal group. In the CCD-986sk cell group treated with zug and camphor extract, the amount of protein expressed by each transcription factor increased in a concentration-dependent manner, and it was confirmed that the expression level of the normal group was recovered at a concentration of 10 μg / mL. Therefore, it was confirmed that the extracts of Zugu and Pseudomonas spp. Promoted the collagen synthesis by activating the damaged procollagenase and showed the effect of promoting the recovery of the damaged epithelium.

(6) Fiber  The wrinkle-related protein (CCD-986sk) MMP -1) and mRNA  Check the expression level

Collagen is a major constituent protein that accounts for 90% of skin dry weight. Therefore, degradation of collagen directly affects the elasticity of the connective tissue and wrinkle formation. In addition, the factor most affected by MAPK, which is an aging-related factor, is c-fos, which is affected by p38. Activation of these factors strongly regulates the expression of MMPs. Among the several MMPs produced in the body, MMP-1 is a protease specifically acting on collagen. It is known that inhibition of MMP-1 activity reduces collagen degradation, maintains elasticity of skin tissue and prevents wrinkle formation .

The activity of MMP-1 in CCD-986sk cells was measured and the results are shown in FIG.

As shown in FIG. 22, it was confirmed that both zug, loose-leaf and alcohol extracts showed inhibition rates in a concentration-dependent manner. In addition, alcohol extracts were more effective in inhibiting the expression of MMP-1 than water extracts, and were similar to those of epigallocatechin-3-O-gallate (EGCG) as a control.

(7) Inhibitory effect of intestitial collagenase (MMP-1) and 92kD gelatinase (MMP-9) on fibroblast (CCD-986sk) protein expression by Western blotting analysis

Reconstruction of extracellular matrix proteins in the dermis involves a variety of biological reactions, and these substrate proteins play an important role in maintaining the structure and resilience of the skin. (MMP-1), 72 kD gelatinase (MMP-2), 92 kD gelatinase (MMP-9), neutrophil collagenase (MMP-8) and collagenase (MMP-9) 13) and more than 26 MMPs are already known. In skin exposed to excessive ultraviolet rays, the expression of MMPs is increased, and the increased MMPs break down the collagen that constitutes the skin, resulting in the deficiency of the substrate protein, thereby causing wrinkles along with aging of the skin.

Inhibition of MMP-1 activity as a protease specifically acting on collagen, which accounts for 90% of skin connective tissues, can reduce collagen degradation, maintain skin elasticity and prevent wrinkle formation. In addition, MMP-9, a 92 kD gelatinase in the same order as 72 kD gelatinase (MMP-2) among enzymes, induces the activity of gelatinase, which causes decomposition of gelatin existing as an intercellular material of the dermal layer. Is an important factor. Therefore, inhibiting the activity of MMP-9 in combination with MMP-1 has been shown to reduce collagen degradation, maintain skin elasticity and prevent wrinkle formation.

Therefore, the expression of MMP-1 and MMP-9 protein in CCD-986sk cells was confirmed by western blotting in order to investigate the effect of extracts of Zug and Pyrite on MMP-1 and MMP-9 protein expression. Β-actin, a house keeping gene, was used as a positive control with little difference in the expression level of the cells under various conditions. The results are shown in Fig.

As shown in FIG. 23, it was confirmed that the expression level of MMP-1 was markedly increased in the control group irradiated with UV-B compared with the normal group. In the CCD-986sk cell group treated with zug and camellia extract, the amount of protein expressed by each transcription factor decreased in a concentration-dependent manner. At a concentration of 10 μg / mL, the inhibitory rate was lower than that of EGCG used in functional cosmetics I could confirm the better. Therefore, it was confirmed that the extracts of waste pomace and mosquito repress the expression of MMP-1 protein, resulting in suppression of collagenase, thereby exhibiting an effect of improving wrinkles and promoting recovery from damaged epithelium I could.

In the case of MMP-9, it was also confirmed that the expression level of the protein in the control group was significantly increased as compared with that in the normal group. In the CCD-986sk cell group treated with the extract of Zugu and Activalrum, protein expression of each transcription factor In the case of 10 μg / mL of water extract, the inhibition rate of EGCG was similar to that of EGCG, which is currently used in functional cosmetics. In alcohol extracts, EGCG is more effective than control EGCG And the like. Therefore, it was confirmed that the extracts of Zug and Sophorae suppressed the expression of MMP-9 protein, and as a result, the wrinkles were improved.

(8) TIMP -1 Active kit measurement result

MMP activation can be specifically inhibited by tissue inhibitors of metallo-proteinase as well as non-specific inhibitors such as α2-macroglobulin and RECK. TIMPs are specific inhibitors that specifically bind 1: 1 in stoichiometry. TIMP-1 is an important inhibitor of MMP-1 in tissues.

In order to measure the activity of TIMP-1, CCD-986sk cells were irradiated with UV-B, and the extracts were treated with 1, 5, and 10 μg / mL concentrations and cultured for 48 hours. The cell culture was taken and the activity of TIMP-1 was measured. The results are shown in FIG.

As shown in FIG. 25, at a treatment concentration of 10 μg / mL, a high expression rate of 85% in the case of the extract of Zug and the seed wool and 97% in the case of the alcohol extract was confirmed. These results indicate that TIMP-1 activity is restored to normal by treatment of zug and scrapie extracts, thereby inhibiting the activity of MMP-1, thereby reducing collagen degradation, maintaining skin elasticity and preventing wrinkle formation .

Formulation example  One. Cosmetics  Manufacture of pharmaceutical preparations

Flexible longevity manufacturing

0.1% by weight of the crude extract of the zodiac in Example 2, 5.2% by weight of 1,3-butylene glycol, 1.5% by weight of oleyl alcohol, 3.2% by weight of ethanol, 3.2% by weight of polysorbate 20, , 1.0% by weight of a carboxyl vinyl polymer, 3.5% by weight of glycerin, a small amount of fragrance, a small amount of preservative and a remaining amount of purified water were mixed to prepare a softening liquid.

Milk lotion  Produce

0.1% by weight of the zug and the actual fraction extract of Example 2, 5.1% by weight of glycerin, 4.2% by weight of propylene glycol, 3.0% by weight of tocopheryl acetate, 4.6% by weight of liquid paraffin, 1.0% by weight of triethanolamine, 3.1% by weight of squalane, 2.5% by weight of polysorbate 60, 1.6% by weight of polysorbate 60, 1.6% by weight of sorbitan sesquioleate, 0.6% by weight of propylparaben, 1.5% by weight of carboxyl vinyl polymer, a small amount of fragrance, a small amount of preservative, To prepare a milk lotion.

Nutrition cream manufacturing

The composition of Example 2 was prepared in the same manner as in Example 2 except that 0.5% by weight of the zug and the actual fraction extracts, 4.0% by weight of glycerin, 3.5% by weight of petroleum jelly, 2.1% by weight of triethanolamine, 5.3% by weight of liquid paraffin, 3.0% by weight of squalane, 2.6% by weight of beeswax, 3.2% by weight of polysorbate 60, 1.0% by weight of a carboxyl vinyl polymer, 3.1% by weight of sorbitan sesquioleate, a small amount of fragrance, a small amount of preservative and a remaining amount of purified water were mixed to prepare a nutritional cream by a conventional method.

Massage cream manufacturing

Example 1 was repeated except that the crude extract of Example 2 and 0.5% by weight of glycerin, 4.0% by weight of glycerin, 3.5% by weight of petroleum jelly, 0.5% by weight of triethanolamine, 24.0% by weight of liquid paraffin, 3.0% by weight of squalane, 2.1% by weight of beeswax, , 2.4% by weight of Polysorbate 60, 1.0% by weight of a carboxyl vinyl polymer, 2.3% by weight of sorbitan sesquioleate, a small amount of fragrance, a small amount of preservative and a remaining amount of purified water were mixed to prepare a massage cream.

Formulation example  2. Manufacture of food preparation

Health food manufacturing

100 mg of the zitta extract of the Example 2, a proper amount of vitamin A, 70 g of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 g of vitamin B12, 10 g of biotin, 1.7 mg of nicotinic acid amide, 50 g of folate, 0.5 mg of calcium pantothenate, 1.75 mg of ferrous sulfate, 0.82 mg of zinc oxide, 25.3 mg of magnesium carbonate, 15 mg of monobasic potassium phosphate, , 90 mg of potassium citrate, 100 mg of calcium carbonate and 24.8 mg of magnesium chloride were mixed and granules were prepared, and a health food was prepared according to a conventional method. At this time, although the composition ratio of the vitamin and mineral mixture is relatively mixed with the ingredient suitable for health food, it may be arbitrarily modified.

Health drink  Produce

100 mg of vitamin C, 100 g of vitamin E (powder), 19.75 g of iron lactate, 3.5 g of zinc oxide, 3.5 g of nicotinic acid amide, 0.2 g of vitamin A, 0.25 g of vitamin B1, 0.3 g of vitamin B2 and a predetermined amount of water were mixed and heated at 85 DEG C for about 1 hour with stirring. The resulting solution was filtered and sterilized in a sterilized 2 L container, . At this time, although the composition ratio of the ingredients suitable for the beverage is comparatively mixed, the mixture ratio may be arbitrarily varied according to the demand, the demanded country, the intended use, and the regional or national preference.

Formulation example  3. Manufacture of pharmaceutical preparations

Powder  Produce

20 mg of the zug and the actual grain extract of Example 2, 100 mg of lactose and 10 mg of talt were mixed and filled in an airtight container to prepare powders by a conventional method.

Tablet manufacture

The tablets were prepared by mixing 10 mg of the zug and obsolute extract of Example 2, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate and then tableting them by a conventional method.

Capsule  Produce

A capsule was prepared by mixing 10 mg of the zug and the actual fraction extract of Example 2, 3 mg of crystalline cellulose, 14.8 mg of lactose and 0.2 mg of magnesium stearate and filling the mixture into gelatin capsules by a conventional method.

Injection manufacturing

(2 mL) per ampoule: 10 mg of the zug and the actual fraction extract of Example 2, 180 mg of mannitol, 2,974 mg of sterile distilled water for injection, and Na ₂ HPO ₄ ? 2H ₂ O (26 mg).

Liquid  Produce

To the purified water was added 20 mg of the crude extract and the false-dead extract of Example 2, 10 g of isomerized sugar and 5 g of mannitol and dissolved, and the lemon flavor was added in an appropriate amount. Then, purified water was further added thereto, adjusted to a total volume of 100 mL, filled in a brown bottle, and sterilized to prepare a liquid preparation by a conventional method.

Claims (7)

A cosmetic composition for whitening, shrinking pores or improving wrinkles comprising an extract of pine nuts and shells as an active ingredient. The cosmetic composition according to claim 1, wherein the pine nuts are mature pine nuts. [Claim 2] The cosmetic composition according to claim 1, wherein the zodiac and bark extracts are extracted with water, C1 to C4 lower alcohols or a mixed solvent thereof. A skin external preparation for whitening, shrinking pores or improving wrinkles, comprising an extract of pine nuts and shells as an active ingredient. The external preparation according to claim 4, wherein the external preparation is selected from the group consisting of a soft lotion, a nutritional lotion, a gel, a cream, an essence, an emulsion, a pack, a cleanser and a powder for a bath. A composition for preventing whitening or aging, which comprises an extract of pine nuts and shells as an active ingredient. The present invention relates to a pharmaceutical composition for the treatment or prevention of a pigmentation disorder comprising an extract of Zygomugovia bark and a skin extract as an active ingredient, wherein the pigmentation disease is freckles, senile spots, melasma, hyperpigmentation after drug use, gravidic chloasma, and post-inflammatory hyperpigmentation due to wound or dermatitis including scratches and burns.

Images