KR102594246B1 - Skin anti-pollution composition containing a novel organic compound as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for skin anti-pollution containing a novel organic compound as an active ingredient, which not only effectively suppresses inflammatory reactions caused by pollutants, but also has excellent antioxidant, anti-aging, and skin whitening improvement effects. A composition for anti-pollution can be provided.
Additionally, cosmetic and food compositions having the above effects can be provided.

Description

Composition for skin anti-pollution containing a novel organic compound as an active ingredient {SKIN ANTI-POLLUTION COMPOSITION CONTAINING A NOVEL ORGANIC COMPOUND AS AN ACTIVE INGREDIENT}

The present invention relates to a composition for skin anti-pollution containing a novel organic compound as an active ingredient. More specifically, it relates to a composition containing a novel organic compound based on natural products that can suppress skin inflammatory reactions that may be caused by fine dust, etc., and also has excellent antioxidant, anti-aging, and skin whitening effects.

As the quality of life has improved due to the extension of average sleep due to recent advances in medical technology, the desire for a healthy life is increasing, and interest in skin care is also increasing.

The skin is a part of the body that is directly exposed to the external environment. It not only acts as a protective layer to protect the important organs of our body, but also regulates moisture evaporation and protects the body from external infections. However, no matter how well the skin blocks virus penetration from the outside, external stress such as excessive ultraviolet rays and a polluted environment causes skin irritation and ultimately leads to skin aging.

As we enter modern society, environmental pollution is worsening and the concentration of pollutants around the living area is increasing, and the issue of protecting the skin from pollutants is emerging as a social issue. Examples of these pollutants include formaldehyde, nitrogen oxides, and benzopyrene, which enter the body and accumulate, causing various diseases.

Meanwhile, one of the causes of skin damage that has been receiving attention in recent years is skin damage caused by fine dust or ultrafine dust. This refers to pollutants with a diameter of 2.5 to 10 ㎛ (PM) or pollutants of 2.5 ㎛ or less, and are known to directly damage the skin barrier function and cause worsening of dermatitis when they come into contact with the skin.

Because the particle size of fine dust is very small, it can penetrate under the skin through hair follicles, so it can affect even those who do not have atopic dermatitis, allergic diseases, or sensitive skin. When fine dust penetrates the skin, it produces active oxygen, damaging the mitochondria of our skin, reducing collagen synthesis, increasing decomposition, and stimulating melanocytes, causing pigmentation, wrinkle formation, and pore expansion. Skin aging may worsen.

Accordingly, various functional cosmetics are being developed, and in particular, investment and research on skin protection cosmetics with anti-pollution functions are actively underway.

Compared to the effects of pollutants mentioned above on the skin, the public's awareness of the risk is still low, but with the recent emergence of issues such as fine dust, the risk is gradually attracting attention, and the number of people who are gradually aware of the risk of pollutants is increasing. The number is increasing.

Accordingly, the development of cosmetic compositions, cleansers, and external agents with anti-pollution effects that prevent environmental pollutants such as those mentioned above from attaching to the surface of the skin or penetrating into the body through the skin is actively underway.

However, the majority of products currently being developed simply remove pollutants from the skin and simply remove contaminants that have adhered to or penetrated the skin. Not only do they not fundamentally protect the skin from environmental pollutants, they also do not fundamentally protect the skin from environmental pollutants. There was a problem that the ability to protect the skin from

In particular, because it contains chemicals as its main ingredient, there is a problem that it acts as a cause of skin troubles and skin diseases. Accordingly, there is a need to develop a composition using natural substances that is mild to the skin and has an excellent anti-pollution effect.

Meanwhile, a bright skin tone symbolizes health and youth, and regardless of age or gender, people's interest in skin whitening is gradually increasing, with people using various cosmetics or undergoing skin treatments to have white, clean skin.

Human skin color is influenced by environment, race, gender, etc., but is generally determined by the content of melanin. Melanin pigment is the biggest factor in determining human skin color, and plays a major role in preventing skin damage from ultraviolet rays by absorbing or scattering ultraviolet rays (Gertrude-E. Costin, et al., The FASED Journal 2001, 21 , 976-994; Yuji Yamaguchi, et al., Journal of Biological Chemistry 2007, 282, 27557-27561).

However, when melanin present in the basal layer of the skin is excessively produced due to various environmental factors such as ultraviolet rays and skin aging, skin pigmentation increases (Iwata M, et al., J Invest Dermatol 1990, 95, 9-15). Diseases that cause abnormalities in skin color include melasma, freckles, and seborrheic keratosis, and this pigmentation causes cosmetic problems, skin aging, and health problems such as skin cancer. FR De Gruijl, European Journal of Cancer 1999, 35, 2003-2009; Bruce K. Armstrong, Anne Kricker, Journal of Photochemistry and Photobiology B: Biology 2001, 63, 8-18).

Ascorbic acid and hydroquinone have already been found to be effective in suppressing the formation of melanin pigment, and are being used as whitening cosmetics. However, due to their poor stability, they are decomposed and discolored, resulting in a minimal whitening effect.

Therefore, the most important issue is developing a whitening material for skin health that can effectively suppress the formation of melanin pigment and is harmless to the human body.

Therefore, in order to solve the above problems, the present applicant confirmed the excellent anti-inflammatory, antioxidant and skin whitening effects of new organic compounds based on natural products and developed a composition for skin anti-pollution containing them as an active ingredient. .

KR 10-1571323 B1 KR 10-1458887 B1

The purpose of the present invention is to provide a composition for skin anti-pollution containing a novel organic compound as an active ingredient.

Another object of the present invention is to provide a composition, which is a novel organic compound based on natural products, that has excellent anti-inflammatory effects against inflammatory reactions caused by pollutants and has excellent antioxidant, anti-aging, and skin whitening effects.

Another object of the present invention is to provide a cosmetic composition and a food composition containing the above composition.

In order to achieve the above object, a composition for skin anti-pollution according to an embodiment of the present invention contains a compound represented by the following Chemical Formula 1 as an active ingredient, and has excellent anti-inflammatory, antioxidant, anti-aging and skin whitening improvement effects. :

[Formula 1]

here,

L ₁ is a single bond or a substituted or unsubstituted alkylene group having 1 to 10 carbon atoms,

Ar ₁ is a substituted or unsubstituted cycloalkyl group with 3 to 30 carbon atoms, a substituted or unsubstituted aryl group with 5 to 30 carbon atoms, a substituted or unsubstituted heteroaryl group with 2 to 30 carbon atoms, or a substituted or unsubstituted heteroaryl group with 3 to 30 carbon atoms. selected from the group consisting of an arylalkyl group and a substituted or unsubstituted aryloxy group having 6 to 30 carbon atoms,

The substituents of L ₁ and Ar ₁ are hydrogen, deuterium, cyano group, nitro group, halogen group, hydroxy group, alkyl group with 1 to 30 carbon atoms, alkenyl group with 2 to 30 carbon atoms, alkynyl group with 2 to 24 carbon atoms, and 2 to 2 carbon atoms. Heteroalkyl group of 30, aralkyl group of 6 to 30 carbon atoms, cycloalkyl group of 3 to 20 carbon atoms, heterocycloalkyl group of 3 to 20 carbon atoms, aryl group of 5 to 30 carbon atoms, heteroaryl group of 2 to 30 carbon atoms, 3 to 3 carbon atoms It is substituted with a substituent selected from the group consisting of a heteroarylalkyl group of 30, an alkoxy group of 1 to 30 carbon atoms, an alkylsilyl group of 1 to 30 carbon atoms, an arylsilyl group of 6 to 30 carbon atoms, and an aryloxy group of 6 to 30 carbon atoms, When substituted with multiple substituents, they may be the same or different from each other.

Ar ₁ may be a compound represented by Formula 2 or Formula 3 below:

[Formula 2]

[Formula 3]

here,

* means the part that is combined.

When measuring ^{a 1} H-NMR spectrum, the compound represented by Formula 1 may exhibit a first peak at 6.86 to 6.88 ppm and a second peak at 6.85 to 6.87 ppm.

The compound represented by Formula 1 may be an active substance isolated and purified from a mountain bone extract.

The compound represented by Formula 1 may be an active substance obtained by fractionating a mountain golchwi extract with n-BuOH and separating and purifying it.

A cosmetic composition for skin anti-pollution according to another embodiment of the present invention may include the above composition.

A food composition for skin anti-pollution according to another embodiment of the present invention may include the above composition.

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In the present invention, “hydrogen” refers to hydrogen, light hydrogen, deuterium, or tritium, unless otherwise specified.

In the present invention, the “halogen group” is fluorine, chlorine, bromine, or iodine.

In the present invention, “alkyl” refers to a monovalent substituent derived from a straight-chain or branched-chain saturated hydrocarbon having 1 to 40 carbon atoms. Examples thereof include methyl, ethyl, propyl, isobutyl, sec-butyl, pentyl, iso-amyl, hexyl, etc., but are not limited thereto.

In the present invention, “alkenyl” refers to a monovalent substituent derived from a straight-chain or branched-chain unsaturated hydrocarbon having 2 to 40 carbon atoms and having one or more carbon-carbon double bonds. Examples thereof include vinyl, allyl, isopropenyl, 2-butenyl, etc., but are not limited thereto.

In the present invention, “alkynyl” refers to a monovalent substituent derived from a straight or branched chain unsaturated hydrocarbon having 2 to 40 carbon atoms and having at least one carbon-carbon triple bond. Examples thereof include ethynyl, 2-propynyl, etc., but are not limited thereto.

In the present invention, “cycloalkyl” refers to a monovalent substituent derived from a monocyclic or polycyclic non-aromatic hydrocarbon having 3 to 40 carbon atoms. Examples of such cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, and adamantine.

In the present invention, “heterocycloalkyl” refers to a monovalent substituent derived from a non-aromatic hydrocarbon having 3 to 40 carbon atoms, and at least one carbon in the ring, preferably 1 to 3 carbons, is N, O, S or Se. It is substituted with a hetero atom such as Examples of such heterocycloalkyl include, but are not limited to, morpholine and piperazine.

In the present invention, “aryl” refers to a monovalent substituent derived from an aromatic hydrocarbon having 6 to 60 carbon atoms, either a single ring or a combination of two or more rings. In addition, a form in which two or more rings are simply attached to each other (pendant) or condensed may also be included. Examples of such aryl include phenyl, naphthyl, phenanthryl, anthryl, fluonyl, dimethylfluorenyl, etc., but are not limited thereto.

In the present invention, “heteroaryl” refers to a monovalent substituent derived from a monoheterocyclic or polyheterocyclic aromatic hydrocarbon having 6 to 30 carbon atoms. At this time, at least one carbon, preferably 1 to 3 carbons, of the ring is replaced with a heteroatom such as N, O, S or Se. In addition, a form in which two or more rings are simply pendant or condensed with each other may be included, and a condensed form with an aryl group may also be included. Examples of such heteroaryls include 6-membered monocyclic rings such as pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, and triazinyl, phenoxathienyl, indolizinyl, and indolyl ( Polycyclic rings such as indolyl, purinyl, quinolyl, benzothiazole, carbazolyl, and 2-furanyl, N-imidazolyl, 2-isoxazolyl , 2-pyridinyl, 2-pyrimidinyl, etc., but are not limited thereto.

In the present invention, “aryloxy” is a monovalent substituent represented by RO-, where R refers to aryl having 6 to 60 carbon atoms. Examples of such aryloxy include phenyloxy, naphthyloxy, diphenyloxy, etc., but are not limited thereto.

As used herein, “aralkyl” refers to an aryl-alkyl group where aryl and alkyl are defined above. Preferred aralkyl contains lower alkyl groups. Non-limiting examples of suitable aralkyl groups include benzyl, 2-phenethyl, and naphthalenylmethyl. Bonding to the parent moiety is via the alkyl.

In the present invention, “alkoxy” may be straight chain, branched chain, or ring chain. The number of carbon atoms of alkoxy is not particularly limited, but is preferably 1 to 20 carbon atoms. Specifically, methoxy, ethoxy, n-propoxy, isopropoxy, i-propyloxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, n-pentyloxy, neopentyloxy, Isopentyloxy, n-hexyloxy, 3,3-dimethylbutyloxy, 2-ethylbutyloxy, n-octyloxy, n-nonyloxy, n-decyloxy, benzyloxy, p-methylbenzyloxy, etc. It may be possible, but it is not limited to this.

In the present invention, “alkylsilyl” refers to silyl substituted with alkyl having 1 to 40 carbon atoms, and “arylsilyl” refers to silyl substituted with aryl having 6 to 60 carbon atoms.

In the present invention, "substitution" means changing a hydrogen atom bonded to a carbon atom of a compound to another substituent. The position to be substituted is not limited as long as it is the position where the hydrogen atom is substituted, that is, a position where the substituent can be substituted, and if two or more substituents are substituted. , two or more substituents may be the same or different from each other. The substituents include hydrogen, cyano group, nitro group, halogen group, hydroxy group, alkyl group with 1 to 30 carbon atoms, alkenyl group with 2 to 30 carbon atoms, alkynyl group with 2 to 24 carbon atoms, heteroalkyl group with 2 to 30 carbon atoms, and 6 to 6 carbon atoms. Aralkyl group of 30, aryl group of 5 to 30 carbon atoms, heteroaryl group of 2 to 30 carbon atoms, heteroarylalkyl group of 3 to 30 carbon atoms, alkoxy group of 1 to 30 carbon atoms, alkylamino group of 1 to 30 carbon atoms, 6 to 6 carbon atoms It may be substituted with one or more substituents selected from the group consisting of an arylamino group of 30, an aralkylamino group of 6 to 30 carbon atoms, and a hetero arylamino group of 2 to 24 carbon atoms, but is not limited to the above examples.

According to the composition for skin anti-pollution containing the novel organic compound of the present invention as an active ingredient, not only can it effectively suppress the inflammatory response caused by pollutants, but it is also a skin anti-pollution agent with excellent antioxidant, anti-aging and skin whitening improvement effects. A composition for solution can be provided. Furthermore, cosmetic and food compositions having the above effects can be provided.

Figure 1 shows the results of ¹ H-NMR analysis of an active material according to an embodiment of the present invention.
Figure 2 shows the results of ¹³ C-NMR analysis of the active material according to an embodiment of the present invention.
Figure 3 shows the results of a cytotoxicity test of a Sangolchwi extract according to an embodiment of the present invention.
Figure 4 shows the results of an experiment showing the inhibitory effect on the inflammatory response induced by fine dust according to an embodiment of the present invention.
Figure 5 shows the results of an experiment showing the inhibitory effect on the inflammatory response induced by fine dust according to an embodiment of the present invention.
Figure 6 shows the results of an experiment on the antioxidant effect according to an embodiment of the present invention.
Figure 7 shows the results of an experiment on the antioxidant effect according to an embodiment of the present invention.
Figure 8 shows the results of an experiment on the antioxidant effect according to an embodiment of the present invention.
Figure 9 shows the results of an experiment on the antioxidant effect according to an embodiment of the present invention.
Figure 10 shows the results of an experiment on the secretion effect of nerve cell active substances according to an embodiment of the present invention.
Figure 11 shows the results of an experiment on the secretion effect of nerve cell active substances according to an embodiment of the present invention.
Figure 12 shows the results of an experiment on the skin whitening effect according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement it. However, the present invention may be implemented in many different forms and is not limited to the embodiments described herein.

In the present invention, “improvement” may mean any action that at least reduces the degree of symptoms, for example, parameters related to the alleviation or treatment of a condition.

In the present invention, “skin anti-pollution” means improving skin contamination, skin cell death or inhibition of skin cell proliferation, or deterioration of skin condition caused by pollutants such as fine dust.

The composition for skin anti-pollution of the present invention contains a compound represented by the following formula (1) as an active ingredient and has excellent anti-inflammatory, antioxidant, anti-aging and skin whitening improvement effects:

[Formula 1]

here,

L ₁ is a single bond or a substituted or unsubstituted alkylene group having 1 to 10 carbon atoms,

Ar ₁ is a substituted or unsubstituted cycloalkyl group with 3 to 30 carbon atoms, a substituted or unsubstituted aryl group with 5 to 30 carbon atoms, a substituted or unsubstituted heteroaryl group with 2 to 30 carbon atoms, or a substituted or unsubstituted heteroaryl group with 3 to 30 carbon atoms. selected from the group consisting of an arylalkyl group and a substituted or unsubstituted aryloxy group having 6 to 30 carbon atoms,

The substituents of L ₁ and Ar ₁ are hydrogen, deuterium, cyano group, nitro group, halogen group, hydroxy group, alkyl group with 1 to 30 carbon atoms, alkenyl group with 2 to 30 carbon atoms, alkynyl group with 2 to 24 carbon atoms, and 2 to 2 carbon atoms. Heteroalkyl group of 30, aralkyl group of 6 to 30 carbon atoms, cycloalkyl group of 3 to 20 carbon atoms, heterocycloalkyl group of 3 to 20 carbon atoms, aryl group of 5 to 30 carbon atoms, heteroaryl group of 2 to 30 carbon atoms, 3 to 3 carbon atoms It is substituted with a substituent selected from the group consisting of a heteroarylalkyl group of 30, an alkoxy group of 1 to 30 carbon atoms, an alkylsilyl group of 1 to 30 carbon atoms, an arylsilyl group of 6 to 30 carbon atoms, and an aryloxy group of 6 to 30 carbon atoms, When substituted with multiple substituents, they may be the same or different from each other.

Specifically, Ar ₁ may be a compound represented by Formula 2 or Formula 3 below:

[Formula 2]

[Formula 3]

here,

* means the part that is combined.

More specifically, the compound represented by Formula 1 may be a compound represented by Formula 4 or Formula 5 below:

[Formula 4]

[Formula 5]

The composition not only has excellent anti-inflammatory effects against inflammatory reactions caused by pollutants, but also has excellent antioxidant, anti-aging, and skin whitening improvement effects.

In addition, the compound represented by Formula 1 may exhibit a first peak at 6.86 to 6.88 ppm and a second peak at 6.85 to 6.87 ppm when measuring a ¹ H-NMR spectrum.

More specifically, the compound represented by Formula 1 shows a first peak at 6.86 to 6.88 ppm, a second peak at 6.85 to 6.87 ppm, and a third peak at 6.58 to 6.60 ppm when measuring the ¹ H-NMR spectrum. peaks, a fourth peak at 5.78 to 5.80 ppm, a fifth peak at 4.56 to 4.58 ppm, and a sixth peak at 3.73 to 3.75 ppm.

Meanwhile, the compound represented by Chemical Formula 1 may be an active substance isolated and purified from a mountain golch extract.

Specifically, the compound represented by Formula 1 may be an active substance obtained by fractionating a mountain golchwi extract with n-BuOH and separating and purifying it.

In other words, the compound represented by Formula 1 is a compound isolated from a fraction of a mountain golchwi extract.

Saussurea neoserrata grows in deep mountains and is 50 to 110 cm tall. The stem stands upright and sometimes produces branches from the upper part. The base of the leaf flows into the stem and becomes the stem's wings. The leaves on the roots and the leaves on the bottom fall off when flowers bloom, and the leaves on the stems are alternate, lanceolate or oval-shaped, and are 12 to 15 cm long. The end is sharp and the bottom is narrow. It gets smaller toward the top, has curly hairs on the back, and has sharp sawtooth edges. Also, the purple flowers bloom in August or September. Head flowers grow in corymbs at the ends of branches and main stems, and the flowers are 8 to 10 mm in diameter. The involucre is barrel-shaped, 8 to 9 mm long and 4 to 5 mm in diameter. The bract pieces are arranged in four rows, have purple edges and have white hairs like spider webs. The fruit is an achene, about 5 mm long, and has black-brown lines. The hairs have two rows and are brown. Young shoots are edible. It is a species endemic to Korea and is distributed in the northern region.

More specifically, the compound represented by Formula 1 can be separated from a fraction fractionated from a hot water extract extracted by a hot water extraction method.

The thermal water extract of Sanggolchwi can be produced using the same method as the production method of a conventional hot water extract.

The thermal water extract of Sangolkchi was partition-extracted with water and ethyl acetate, and the water layer was partition-extracted again with n-butanol. Each layer was concentrated under reduced pressure to prepare EtOAc, n-BuOH, and H ₂ O fractions.

The compound represented by Formula 1 was identified from the butanol fraction of mountain golchwi, and was analyzed through the NMR data shown in Figures 1 and 2.

When measuring ^{a 1} H-NMR spectrum, the compound represented by Formula 1 shows a first peak at 6.86 to 6.88 ppm, a second peak at 6.85 to 6.87 ppm, and a third peak at 6.58 to 6.60 ppm, It may be said to correspond to a compound characterized by exhibiting a fourth peak at 5.78 to 5.80 ppm, a fifth peak at 4.56 to 4.58 ppm, and a sixth peak at 3.73 to 3.75 ppm.

A cosmetic composition for skin anti-pollution according to another embodiment of the present invention may include the above composition.

'Cosmetics' as used in the present invention is not limited to cosmetics and is defined to include all pharmaceuticals or quasi-drug external preparations.

The cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the field, for example, solution, suspension, paste, gel, cream, lotion, powder, soap, surfactant-containing clenosis, oil. , may be formulated as powder foundation, emulsion foundation, wax foundation, and spray, but is not limited thereto. More specifically, flexible lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation, oil-in-water (O/W) type or water-in-oil type. It can be prepared as a (W/O) type formulation or ointment.

The cosmetics can be prepared in the form of supplements that can be applied topically or systemically, as commonly used in the art, by containing a dermatologically acceptable medium or base in addition to the extract of mountain lanceolate of the present invention.

Suitable cosmetic formulations include, for example, solutions, gels, solid or pasty anhydrous products, emulsions obtained by dispersing the oil phase in the aqueous phase, suspensions, microemulsions, microcapsules, microgranules or in ionic (liposome) or non-ionic form. It may be provided in the form of a vesicular dispersion, cream, skin, lotion, powder, ointment, spray or conceal stick. Additionally, it can be prepared in the form of foam or in the form of an aerosol composition further containing compressed propellant.

Products to which the cosmetic composition of the present invention can be added include, for example, cosmetics such as astringent lotion, softening lotion, nourishing lotion, various creams, essences, packs, foundations, etc., and cleansers, face washes, soaps, treatments, or serums. etc. Specific formulations include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, essence, nutritional essence, pack, soap, shampoo, and cleansing foam. , cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, pressed powder, loose powder or eye shadow.

A food composition for skin anti-pollution according to another embodiment of the present invention may include the above composition.

'Food' as used in the present invention includes not only general foods and beverages, but also functional foods or health functional foods.

At this time, the amount of the extract in the food or beverage can be added at about 0.01 to 15% by weight of the total weight of the food, and the health drink composition can be added at a rate of about 0.01 to 10g based on 100 ml, but is not limited thereto. .

When the food composition of the present invention is a beverage composition, other than containing the extract as an essential ingredient in the ratio indicated, there are no particular restrictions on other ingredients, and various flavoring agents or natural carbohydrates may be contained as additional ingredients like ordinary beverages. You can. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used, but are not limited to these. The ratio of the natural carbohydrate may generally range from about 1 to 20 g per 100 ml of the composition of the present invention, but may not be limited thereto.

In addition to the above, the composition of the present invention contains various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its It may contain salt, organic acid, protective colloidal thickener, pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonating agent used in carbonated beverages, etc., but may not be limited thereto.

In addition, the composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks, and these ingredients can be used independently or in combination. The proportion of these additives is not critical, but may be selected in the range of about 0.1 to about 20 parts by weight per 100 parts by weight of the composition of the present invention, but is not limited thereto.

“Functional food” as used in the present invention refers to food manufactured and processed using specific ingredients as raw materials or specific ingredients contained in food raw materials for the purpose of health supplementation, including extracting, concentrating, refining, mixing, etc., and includes health supplements. In addition, it is a food designed and processed to fully exert the bioregulatory functions of food ingredients, such as biodefense, regulation of biorhythm, and disease prevention and recovery, to the living body, and functions related to disease prevention and disease recovery. It is also assumed to include everything that it has.

The above health functional food may additionally contain food additives, and its suitability as a “food additive” is determined by the specifications for the relevant item in accordance with the general provisions and general test methods of the Food Additives Code approved by the Food and Drug Safety Administration, unless otherwise specified. and standards.

Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as pigment reduction, plant extracts, crystalline cellulose, cold pigment, and guar gum, L -Mixed preparations such as sodium glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations are included.

Carriers, excipients, and diluents that may be included in the functional food or health functional food include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, Examples include calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract and its fractions with at least one excipient such as starch, calcium carbonate, It is prepared by mixing sucrose, lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups, and may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives in addition to the commonly used simple diluents such as water and liquid paraffin. there is. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used.

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[Preparation example: Preparation of mountain bone extract]

The dried and pulverized mountain bonech was subjected to hot water extraction and then filtered to prepare a mountain bonech extract, concentrated under reduced pressure at 45°C, and the concentrate was partitioned and extracted with water and ethyl acetate, and the water layer was again treated with n-butanol. Distribution extraction was performed. Each layer was concentrated under reduced pressure to prepare EtOAc, n-BuOH, and H ₂ O fractions.

The n-BuOH fraction was subjected to silica gel column (ϕ5×20 cm) chromatography (CHCl ₃ -MeOH = 1:9) to obtain the fraction. Among these, silica gel column chromatography (CHCl ₃ -MeOH = 1:9, ϕ 3 x 20 cm) was performed on the 2fr. (82 mg) fraction to obtain a fraction. Afterwards, the small fractions were separated and purified. The separated and purified subfraction was purified by Sephadex LH-20 column chromatography (CHCl ₃ :MeOH = 1:9, ϕ2.5×50 cm) to isolate Compound 1 (10.4 mg).

[Experimental Example 1: Analysis of active substances isolated from mountain golchwim]

The active ingredient was analyzed through ¹ H-NMR and ¹³ C-NMR analysis. Analysis was performed using Bruker AVANCE II 400 ( ¹ H NMR at 400 MHz, ¹³ C NMR at 100 MHz).

The analysis results are shown in Figures 1 and 2.

¹ H-NMR spectrum measurement results, δ _H 6.87 (2H, br.s, H-2,6), δ _H 6.86 (1H, d, J= 15.6Hz, H-7) and δ _H 6.59 (1H, dt) , J = 15.6, 4.8Hz, H-8), the presence of one symmetric tetra-substituted aromatic ring and one trans-double bond was confirmed.

In addition, the presence of one sugar molecule was confirmed at _δH 5.79 (1H, d, J= 6.4Hz, H-1'), and oxygenated methylene was found at _δH 4.57 (1H, d, J= 3.8Hz, H-9). The signal was confirmed, and the methoxy signal was confirmed at δ _H 3.74 (6H, s, H-OCH ₃ ). Through this, the above compound was predicted to be an aromatic glycoside derivative.

Meanwhile, 17 carbon signals were confirmed in the ¹³ C-NMR spectrum, including one sugar molecule and two methoxy groups.

Oxygenated olefin quaternary carbon was confirmed in the signal of δc 154.3 (C-3,5), δc 136.4 (C-4), δc 134.4 (C-3), δc 131.6 (C-1), δc 129.8 (C-6) ) and the signal of δc 105.6 (C-2,6) confirmed the presence of one symmetric tetra-substituted aromatic ring and one trans-double bond.

The anomer carbon originating from carbon number 1 of the sugar was confirmed in the signal of δc 105.3 (C-1'), δc 79.2 (C-3'), δc 78.8 (C-5'), δc 76.5 (C-2') , the structure of the sugar was determined to be glucose at the sugar moiety signal of δc 72.0 (C-4') and δc 63.0 (C-6').

In addition, one oxygenated methylene and two methoxy groups were confirmed through signals of δc 63.2 (C-9) and δc 57.0 (C-OCH ₃ ).

Based on the above analysis results, it was analyzed that Syringin, a compound represented by the following formula 4, is the active substance:

[Formula 4]

[Experimental Example 2: Materials and Reagents]

1. Sample

Fine dust (urban particulate matter) used as a pollutant had an average particle size of 10 μm or less and was collected from urban air pollution sources (PM10, Sigma-Aldrich, Darmstadt, Germany). As natural products, sea bamboo shoots and mountain golchwi obtained through hot water extraction were used, and the green leaves were provided by Kang Cosmetique. The compound isolated in Experimental Example 1 was used as a substance for identifying mountain bone odor, and salidroside, an analogue, was purchased and used.

2. Cell lines

The HaCaT, Raw 264.7, Melan-a, and HDF cell lines used in this experiment were purchased from the Korean Cell Line Bank, and were grown in DMEM medium containing 1% antibiotic-antimycotic and 10% FBS for 37 days. °C and cultured in a CO ₂ incubator adjusted to 5% CO ₂ .

[Experimental Example 3: Cytotoxicity experiment]

1. Measurement of cytotoxicity (LDH assay)

Cytotoxicity for HaCaT cells was measured using a non-radioactive cytotoxicity assay using lactase dehydrogenase (LDH) release, and Raw264.7 cells and HaCaT cells were seeded at 1×10 ⁵ cells/in a 48-well plate (SPL, Korea). It was inoculated into a well, and after 18 hours of culture, the hot water extract of Sangolkchi was simultaneously treated at different concentrations and cultured for 24 hours.

45 minutes before recovering the culture medium, control cells were treated with 1×lysis buffer (Promega, USA) and incubated for 45 minutes, and then the culture medium was centrifuged at 3,000 rpm for 5 minutes. 50 μL of culture medium obtained by centrifugation and 50 μL of reconstituted substrate mix (Promega, USA) were added, reacted at room temperature for 30 minutes, and then 50 μL of stop solution (Promega, USA) was added to stop the reaction. Absorbance was measured at 490 nm using a microplate reader.

The average absorbance value for each sample group was obtained, and cytotoxicity was measured by comparing it with the absorbance value of the control group.

The results of the experiment are shown in Figure 3, and it was confirmed that there was no cytotoxicity of the Sangolchwi hot water extract.

[Experimental Example 4: Anti-inflammatory effect experiment on inflammatory response caused by fine dust]

The effect of microparticle PM10 on inflammation was measured by testing the effect of prostaglandin E2 (PGE ₂ ) expression. Epithelial keratinocyte HaCaT cells were inoculated into a 48-well plate at 1×10 ⁵ cells/well, and after 18 hours of culture, fine particles PM10 were treated at different concentrations and cultured for 24 hours. 45 minutes before recovering the culture medium, control cells were treated with 1×lysis buffer (Promega, USA) and incubated for 45 minutes, and then the culture medium was centrifuged at 3,000 rpm for 5 minutes. The expression level of PGE ₂ was measured in the obtained supernatant using a PGE ₂ ELISA assay kit (R&D system, USA). All samples were stored frozen below -20°C until quantification.

Figure 4 confirms the effect of inducing prostaglandin E2 (PGE ₂ ) expression in the inflammation effect caused by fine particles PM10, and it was confirmed that the effect of inducing PGE ₂ expression was highest at 25 μg/mL.

Using the above 25 μg/mL as a control, Saussurea neo serrata, Syringin, and Salidroside were treated at 1 μg/mL and 2 μg/mL, respectively, to confirm the PGE ₂ inhibition effect by fine particles PM10.

The experimental results are shown in Figure 5, and it was confirmed that the PGE ₂ inhibitory effect was superior when treated alone with Syringin and Salidroside, the active substances isolated in Experimental Example 1, compared to the control group.

In light of the above experimental results, it can be said that the active substance isolated from mountain golchwi has an excellent inhibitory effect on inflammatory mediators caused by fine dust.

[Experimental Example 5: Intracellular antioxidant effect experiment]

1. Ultraviolet B irradiation

UVB irradiation is performed after removing the culture medium, washing HaCaT cells with PBS, and adding enough PBS to slightly cover the cells. Cells were placed in a UV irradiator and irradiated with a wavelength of 275 to 380 nm (maximum 290 to 320 nm) using a TL 20W/12 RS fluorescent sun lamp (Philips, Netherlands). At this time, a TA 401/407 Kodacel filter (Kodak, USA) was used to remove UVC from the lamp, and the intensity of UV irradiated to the cells was measured with a UV meter (Ultra-Violet Products Ltd., UVP, USA). did.

2. Measurement of the effect of suppressing intracellular oxygen radical production (hydroxyl radical)

To measure the effect of UVB irradiation on the inhibition of intracellular hydroxyl radical production in HaCaT cells, UVB (100 mJ/cm ² ) was irradiated for 130 min and then the fluorescent probe 2',7'-dichlorofluorescein diacetate (DCF-DA: Changes in the concentration of hydroxyl radicals in cells were measured using a molecular probe.

DCF-DA, a non-fluorescent substance, reacts with hydroxyl radicals and hydrogen peroxide in cells and is converted to fluorescent DCF, which emits green fluorescence. ROS can be measured using DCF-DA. 1 × 10 ⁴ HaCaT cells were inoculated into a 96-well plate (SPL, Korea) and cultured for 18 hours, and simultaneously treated at each concentration and cultured for 24 hours. After completion of incubation, the medium was removed and washed twice with PBS. 10 μM DCF-DA was added to each well and incubated for 90 minutes in a 5% CO ₂ incubator at 37°C. Then, the fluorescence intensity was measured through flow cytometry, and the fluorescence intensity was measured using a laser fluorescence microscope (LSM 510, Carl Zeiss, Germany). At 400 magnification, the excitation wavelength was observed to be 488 nm (argon laser) and the emission wavelength was observed to be 505 nm. After drug administration, changes in fluorescence intensity within cells were observed.

Syringin and Salidroside, the active substances isolated from the mountain golch extract, were treated at 1 mg/mL and 2 mg/mL, respectively, and their efficacy was compared and analyzed using the same method as the experiment on the inhibition effect of PGE ₂ by fine particles PM10.

The experimental results are shown in Figures 6 to 9. Figure 6 shows the change in fluorescence intensity due to UVB irradiation as a control group, and then tested the effect of inhibiting the production of active oxygen radicals for each.

As a result of the experiment, it was confirmed that Syringin and Salidroside, which are active substances isolated from mountain golchwi extract, had a superior effect on suppressing oxygen radical production compared to the control group.

[Experimental Example 6: Nerve cell active substance secretion experiment]

1. Cell culture

Human-derived skin fibroblasts used the HDF cell line, and were cultured in DMEM medium containing 0% FBS, 100 mg/L streptomycin, 100,000 U/L penicillin, in a 10% CO ₂ incubator at 37°C.

2. Extraction sample processing and UVB irradiation

After stabilizing the cells by distributing them in a 6-well plate at a density of ⁵ was added and then UVB was irradiated.

Cells were placed in a UV irradiator and irradiated with a wavelength of 275 to 380 nm (maximum 290 to 320 nm) using a TL 20W/12 RS fluorescent sun lamp (Philips, Netherlands). At this time, a TA 401/407 Kodacel filter (Kodak, USA) was used to remove UVC from the lamp, and the intensity of UV irradiated to the cells was measured with a UV meter (Ultra-Violet Products Ltd., UVP, USA). did.

3. Effects on survival and development of nerve cells

HDF cells were treated with the extract sample and UVB, cultured for 48 hours, then the culture medium was collected, only the supernatant was collected, and the production concentration of nerve growth factor-β and neurotrophin-3 was measured using an enzyme linked immunosorbent assay (ELISA) kit.

The experimental results are shown in Figures 10 and 11.

As a result of the experiment, it was confirmed that Syringin and Salidroside, the active substances isolated from the extract of mountain golchwi, showed higher production concentrations of nerve growth factor-β and neurotrophin-3 compared to the control group.

[Experimental Example 7: Whitening efficacy experiment]

1. Cell culture

Melan-a cells are an immortalized melanocyte cell line and were cultured in 10% FBS, 200 nM TPA, 100 mg/L streptomycin, 100,000 U/L penicillin, and RPMI medium in a 10% CO ₂ incubator at 37°C.

2. Melanin production inhibition test in melanocytes

Dispense to a 6-well plate at a concentration of ² Absorbance was measured in nm, and the efficacy of melanin production inhibition in the sample treatment group was expressed as a ratio to the control value.

The experimental results are as shown in Figure 12, and compared to Arbutin, which is known to have an excellent effect on skin whitening, it can be confirmed that the active substance isolated from mountain golch exhibits an equal or superior melanin production inhibition effect, showing an excellent whitening activity effect. You can see that there is.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims are also possible. falls within the scope of rights.

Claims (8)

Contains a compound represented by the following formula 4 as an active ingredient,
The compound represented by the following formula 4 is separated and purified from the n-BuOH fraction of the thermal water extract of Sangolkchi,
Excellent skin whitening improvement effect
Composition for improving skin whitening:
[Formula 4]
delete delete delete delete Comprising a composition according to claim 1
Cosmetic composition for improving skin whitening. Comprising a composition according to claim 1
Food composition for improving skin whitening. delete