Cosmetic composition for pore improvement, skin barrier improvement, wrinkle improvement and glycosylation resistance containing artichoke extract having improved 1, 5-dicaffeoylquinic acid and chlorogenic acid

Abstract

The present invention relates to an anti-glycosylation composition for improving pores, improving skin barrier, improving wrinkles and resisting glycosylation, containing an artichoke extract in which 1, 5-dicaffeoylquinic acid and chlorogenic acid are increased. According to the artichoke, the state that the content of 1, 5-dicaffeoylquinic acid and chlorogenic acid in the extract is increased through raw material pretreatment engineering is achieved, and the pore size reducing effect and the sebum adjusting effect of the extract are remarkably improved; in addition, various skin health improving effects, skin barrier improving effects and the like, which are obtained by anti-glycosylation effects, are more excellent than those of the conventional artichoke extract, thereby exhibiting suitability of a highly functional cosmetic composition which can improve sensitivity and complex skin.

Description

Technical Field

[0001] This invention relates to a cosmetic composition for improving pores, skin barrier, wrinkles, and anti-glycation of artichoke (Cynarascolymus) extract containing enhanced 1,5-dicaffeoylquinic acid and chlorogenic acid, characterized in that the artichoke is pretreated by low-temperature vacuum drying. Background Technology

[0002] As an organ in contact with the external environment, the skin protects the body from physical damage and chemical substances, prevents bacteria, fungi, and viruses from penetrating the body, and acts as a barrier to prevent moisture loss. Furthermore, the skin consists of three layers: the epidermis, dermis, and subcutaneous fat. The epidermis, as the thinnest layer, plays a crucial protective role. Generally, collagen and elastin, which make up the majority of the dermis, are the main proteins produced by fibroblasts. Deep within the dermis, they participate in aspects such as the skin's mechanical strength, tissue cohesion, and elasticity. Research on the genes involved in their synthesis is also actively underway.

[0003] Glycation, a non-enzymatic reaction between sugars, proteins, and lipids, involves the reaction of the carbonyl group of a reducing sugar with the free amino group of a protein to form Schiff bases, Amadori products, and Maillard products. The compounds formed in this manner can then undergo a series of reactions to generate brown, irreversible advanced glycation end products (AGEs). These AGEs can lead to darkening of the skin and induce dryness and wrinkles. Therefore, the accumulation of AGEs has become a major concern in recent years as a primary cause of endogenous aging. In the Chinese beauty market, cosmetics targeting anti-glycation rather than anti-oxidation have also gained significant attention.

[0004] Keratinocytes, as specialized cells that continuously proliferate from the basal cells at the bottom of the epidermis, undergo phased morphological and functional changes before rising to the skin surface. Over time, the oldest keratinocytes shed from the skin, replaced by new ones. This recurring series of changes is called "epidermal cell differentiation" or "keratinization." During keratinization, keratinocytes form the stratum corneum by producing natural moisturizing factor (NMF) and intercellular lipids (ceramides, cholesterol, fatty acids), giving the stratum corneum its strength and flexibility, thus enabling it to function as a skin barrier. NMF functions as a moisture reservoir in the stratum corneum. After the production of a protein called filaggrin in skin cells, filaggrin is converted into NMF in the stratum corneum. The increased synthesis of filaggrin, as described above, directly represents an increase in NMF, thus implying improved moisturizing function. Furthermore, the moisture within the stratum corneum helps maintain the activity of various physiologically active enzymes and also contributes to maintaining a healthy skin barrier. When the skin's moisture content decreases, it can damage the skin barrier, ultimately leading to decreased elasticity and wrinkle formation. In other words, the loss of skin's moisturizing ability is a cause of reduced skin elasticity.

[0005] In order to solve the problems that occur on the skin as described above, there is a need to develop new and useful substances from natural substances whose safety has been ensured, and therefore related research activities are being actively carried out.

[0006] Artichokes are perennial plants belonging to the Asteraceae family, native to the Mediterranean coast. Their heads are shaped like a receptacle surrounded by bracts, and their stamens are mainly used for food.

[0007] 1,5-Dicaffeoylquinic acid, a naturally occurring polyphenolic compound, has been reported to possess antioxidant and anti-inflammatory properties; however, research on the efficacy of 1,5-dicaffeoylquinic acid in artichokes is still insufficient. Therefore, this invention confirms that artichoke extract possesses excellent sebum / pore regulation, skin barrier improvement, wrinkle / elasticity improvement, and anti-glycation activity, thereby providing a cosmetic composition for skin improvement containing said artichoke extract.

[0008] Prior technology documents

[0009] Patent documents

[0010] (Patent Document 0001) Korean Patent Publication No. 10-2024-0082416 (Invention Title: Anti-atopic, anti-acne or anti-pruritus composition containing caffeoylquinic acid as an active ingredient; Applicant: FIFTEEN D GRIZ Co., Ltd.; Publication Date: 2024.06.11)

[0011] (Patent Document 0002) Korean Patent Registration No. 10-2335297 (Invention Title: Cosmetic Composition for Relieving or Soothing Skin from External Stimuli by Compound Extracts of Artichoke Leaf, Camellia Leaf and Capernaum Fruit Treated under Ultra-High Pressure, Applicant: Hyundai Broland Co., Ltd., Registration Date: 2021.12.01)

[0012] (Patent Document 0003) Korean Patent Registration No. 10-2197684 (Invention Title: Composition for Strengthening Skin Barrier and Preventing Skin Barrier Damage Containing 3,5-Dicaffeoylquinic Acid as an Active Ingredient; Applicant: Korea University Industry-Academia Collaboration Group; Registration Date: 2020.12.24)

[0013] (Patent Document 0004) Korean Patent Registration No. 10-2154927 (Invention Title: Functional Cosmetic Composition for Skin Improvement Containing Chlorogenic Acid, Ferulic Acid, Resveratrol and Streptococcus thermophilus Fermentation as Active Ingredients, Applicant: Theranostics Co., Ltd., Registration Date: 2020.09.04)

[0014] (Patent Document 0005) Chinese Patent Publication No. 101691330 (Invention Title: Method for Separation and Purification of High-Purity Antiviral Active Components from Artichokes, Applicant: Central South University, Publication Date: 2010.04.07) Summary of the Invention

[0015] The purpose of this invention is to provide a cosmetic composition for improving pores, skin barrier, wrinkles, and anti-glycation of an artichoke extract containing enhanced 1,5-dicaffeoylquinic acid and chlorogenic acid, characterized in that the artichoke is pretreated by low-temperature vacuum drying.

[0016] This invention relates to an anti-glycation cosmetic composition containing an artichoke extract with enhanced 1,5-dicaffeoylquinic acid and chlorogenic acid.

[0017] The artichokes can be dried at 4 to 37°C and under low-temperature vacuum conditions of 0 to -760 mmHg, preferably at 4 to 20°C and under low-temperature vacuum conditions of -700 to -760 mmHg. Furthermore, the drying time should preferably be maintained at 8 to 48 hours.

[0018] The artichoke extract can be extracted using a 20-50% (v / v) aqueous solution of 1,3-butanediol as a solvent.

[0019] The artichoke extract may contain 2.8 to 4.0% by weight of 1,5-dicaffeoylquinic acid and 2.8 to 4.0% by weight of chlorogenic acid.

[0020] The extract has the effect of improving pores or regulating sebum, preferably reducing the size and number of skin pores and sebum secretion.

[0021] The extract may have skin barrier improving effects, preferably enhancing the gene expression of hyaluronic acid synthase 3 (HAS3), caspase 14 (CAS14), filaggrin (FLG) or ceramide synthase 3 (CERS3).

[0022] In addition, the extract may have wrinkle-improving or elasticity-improving effects, preferably enhancing the gene expression of type I collagen (COL1A1), type III collagen (COL3A1), or elastin.

[0023] The present invention will now be described in detail.

[0024] The extract of the present invention, at a concentration of 100 to 150 μg / ml, can enhance the gene expression of type I collagen (COL1A1) by 180 to 260%, type III collagen (COL3A1) by 120 to 170%, and elastin by 120 to 180%. Furthermore, it can enhance the gene expression of hyaluronic acid synthase 3 (HAS3) by 150 to 220%, caspase 14 (CAS14) by 125 to 180%, filaggrin (FLG) by 135 to 180%, and ceramide synthase 3 (CERS3) by 125 to 180%.

[0025] The extract of the present invention is characterized by having a glycosylation inhibition rate of more than 40%, preferably 40-65%, at 100 to 150 μg / ml.

[0026] The extract of the present invention also has the effects of reducing sebum by 25 to 30%, reducing pore size by 28 to 33%, and reducing wrinkle depth by 16 to 20%.

[0027] The extract can be obtained by extracting artichoke raw material that has undergone low-temperature vacuum drying pretreatment using water, C1-C4 alcohols, 1,3-butanediol, or a mixture thereof as solvents. The C1-C4 alcohols can be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, and isobutanol. The extract can also be an aqueous extract of 20 to 90% (v / v) alcohol or 1,3-butanediol, preferably 20 to 50% (v / v) alcohol or 1,3-butanediol, and more preferably 20 to 50% (v / v) 1,3-butanediol.

[0028] As the solvent used in manufacturing the extract, 1 to 40 times the volume (1 to 40 l based on 1 kg) or 1 to 40 times the weight of the raw material sample can be used, preferably 5 to 40 times the volume or 5 to 40 times the weight, and more preferably 10 to 40 times the volume or 10 to 40 times the weight. The extraction conditions for the extract can be at 40 to 90°C for 3 to 96 hours, preferably at 40 to 60°C for 3 to 48 hours. The process can be repeated 1 to 4 times.

[0029] After extraction, the material can be pre-treated and filtered using materials such as non-woven fabric, paper, and cloth, or filtered using filters with different pore sizes, such as filters with pore sizes of 10, 5, 2, 1, 0.6, 0.45, and 0.2 μm. Other filters can be used, added, or omitted as needed.

[0030] In addition, the extract can also be extracted by adding buffer solutions such as physiological saline, tris(hydroxymethyl)aminomethane hydrochloride (Tri-HCl), physiological saline solution, phosphate buffer (PBS), and Hank's balanced salt solution (HBSS) to the raw material sample; buffer solutions containing one or more of the following: arginine, vitamin C, tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl), glycine, bisphosphonate, phosphate, potassium dihydrogen phosphate, and potassium phosphate; or buffer solutions containing ionized components such as calcium, iodine, iron, magnesium, selenium, zinc, sodium, phosphorus, sulfur, chlorine, copper, manganese, iron, and guanidine.

[0031] As the extraction equipment for the extract, general extraction equipment, ultrasonic pulverizers, or fractionators can be used. The extract produced by the method described above can be solvent-removed by hot air drying, vacuum drying, or freeze-drying. Furthermore, the extract can be used after purification using column chromatography.

[0032] The extracts described above can be used after fractionation or purification using conventional methods, either alone or in appropriate combinations, known methods used in the separation and extraction of plant components, such as extraction with organic solvents (alcohol, ether, acetone, etc.), partitioning with hexane and water, and column chromatography.

[0033] The chromatographic method can be selected from silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium-pressure liquid chromatography, thin-layer chromatography (TLC), silica gel vacuum liquid chromatography, and high-performance liquid chromatography.

[0034] Furthermore, the present invention provides a skin-improving pharmaceutical composition containing an extract of artichoke pretreated by low-temperature vacuum drying and a pharmaceutical excipient. In the pharmaceutical composition of the present invention, the extract may be added at 0.001–30% by weight.

[0035] The pharmaceutical composition can be formulated into oral dosage forms, external dosage forms, suppositories, and sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, using conventional methods. Carriers, excipients, and diluents that can be included in the pharmaceutical composition may include, for example, lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. During formulation, commonly used fillers, expanders, binders, humectants, disintegrants, and surfactants are used as diluents or excipients. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules. As described above, these solid dosage forms are prepared by mixing the extract of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid dosage forms for oral administration include suspensions, internal solutions, emulsions, and syrups. Besides commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as humectants, sweeteners, flavorings, and preservatives. Dosage forms for non-oral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Non-aqueous solvents and suspensions can use vegetable oils such as propylene glycol, polyethylene glycol, and olive oil, as well as injectable esters such as ethyl oleate. As a base for suppositories, materials such as Witepsol, polyethylene glycol, macrogol, Tween 61, cocoa ester, lauryl ester, and glycerin gelatin can be used.

[0036] The dosage of the pharmaceutical composition of the present invention may vary depending on the age, sex, weight, specific disease or pathological condition requiring treatment, severity of the disease or pathological condition, route of administration, and the prescribing physician's judgment of the recipient. Determining the dosage based on these factors is within the competence of a person skilled in the art, and generally, the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. More preferably, the dosage can be from 1 mg / kg / day to 500 mg / kg / day. Administration may be once daily or multiple times daily. All aspects of the dosage are not intended to limit the scope of the invention.

[0037] The pharmaceutical compositions of the present invention can be administered to mammals such as mice, livestock, and humans via a variety of routes. All routes of administration are predictable, including oral, rectal or intravenous, intramuscular, intradural, or intracerebral injection. The compositions of the present invention have virtually no toxicity or side effects, and therefore can be used safely even for prolonged use as a preventative measure.

[0038] As a dosage form of the cosmetic composition, it can be manufactured in any dosage form commonly manufactured in the industry, such as serums, toners, creams, lotions, masks, hand creams, foot creams, body lotions, lip balms, lipsticks, eyeshadows, eyeliners, eyebrow pencils, blushes, highlighters, regular toners, lotions, creams, serums, beauty soaps, softening lotions, medicated toners, body cleansers, facial foams, cleansing milks, gels, cleansing oils, cleansing creams, cleansing wipes, cleansing waters, masks, and various other hair care products, especially those used in... When it comes to various hair cosmetic products such as those for hair growth, improving hair loss, and improving scalp condition, the formulation is not subject to any particular limitation. More specifically, you can choose from any formulation such as shampoo bars, shampoos, conditioners, hair serums, hair treatments, hair essences, hair toners, hair growth serums, hair lotions, hair creams, hair massage creams, hair waxes, hair masks, hair oils, hair dryers, hair conditioning treatments, hair dyes, hair bleaching agents, hair gels, hair shine products, mousses, hair sprays, and hair ampoules.

[0039] More specifically, when the cosmetic composition of the present invention is in the form of a paste, cream, or gel, carrier components such as animal oils, vegetable oils, waxes, paraffin wax, starch, astragalus gum, cellulose derivatives, polyethylene glycol, organosilicon, bentonite, silica, talc, or zinc oxide can be used. When the cosmetic composition of the present invention is in the form of a powder or spray, carrier components such as lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used. Especially in the case of a spray, propellants such as chlorofluorocarbons, propane-butane, or dimethyl ether can be added. When the cosmetic composition of the present invention is in the form of a solution or emulsion, carrier components such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol oil, glycerol aliphatic esters, polyethylene glycol, or dehydrated sorbitol fatty acid esters can be used as carrier components, including solvents, solubilizers, or emulsifiers. When the cosmetic composition of the present invention is in the form of a suspension, liquid diluents such as water, ethanol, or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol and polyoxyethylene sorbitol ester, microcrystalline cellulose, aluminum hydroxide, bentonite, agar, or xanthan gum can be used as carrier components. When the cosmetic composition of the present invention is in the form of a surfactant-containing detergent, carrier components such as fatty alcohol sulfates, fatty alcohol ether sulfates, sulfosuccinate monoesters, acetoacetate, imidazolinetonium derivatives, methyl taurate, sarcosinate, fatty acid amide ether sulfates, alkylamide betaine, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic acid derivatives, or ethoxylated glycerol fatty acid esters can be added. The cosmetic composition of the present invention may also contain excipients including fluorescent substances, fungicides, hydrotropic inducers, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, opacifiers, and plant extracts. The amount of the ingredient added may be selected in a range that does not impair the inherent effects of the cosmetic composition, depending on the dosage form or intended use. The amount of the ingredient added may be from 0.1 to 10% by weight relative to the total weight of the composition, preferably from 0.1 to 6% by weight, but is not limited thereto.

[0040] This invention relates to an anti-glycation composition containing artichoke extract with enhanced 1,5-dicaffeoylquinic acid and chlorogenic acid, for improving pore size, skin barrier function, wrinkle reduction, and anti-glycation effects. The artichoke extract is prepared by pre-processing the raw material to increase the content of 1,5-dicaffeoylquinic acid and chlorogenic acid. The extract exhibits significantly improved pore size reduction and sebum regulation effects, and its various skin health improvement and skin barrier enhancement effects are superior to existing artichoke extracts, demonstrating its suitability as a high-functionality cosmetic composition for improving sensitive and complex skin. Attached Figure Description

[0041] Figure 1 illustrates the high-performance liquid chromatography (HPLC) results of the extracts of Example 1 (a), Example 2 (b), Example 3 (c), Comparative Example 1 (d), Comparative Example 2 (e), Comparative Example 3 (f), and Comparative Example 4 (g).

[0042] Figure 2 The results confirming the expression of type I collagen (COL1A1) gene in the artichoke extract of the present invention are illustrated.

[0043] Figure 3 The results confirming the expression of type III collagen (COL3A1) gene in the artichoke extract of the present invention are illustrated.

[0044] Figure 4 The results of confirming the expression of the elastin gene in the artichoke extract of the present invention are illustrated.

[0045] Figure 5 The results of confirming the expression of the hyaluronic acid synthase 3 (HAS3) gene in the artichoke extract of the present invention are illustrated.

[0046] Figure 6 The results confirming the expression of the caspasin 14 (CAS14) gene in the artichoke extract of the present invention are illustrated.

[0047] Figure 7 The results confirming the expression of the filaggrin (FLG) gene in the artichoke extract of the present invention are illustrated.

[0048] Figure 8 The results confirming the expression of the ceramide synthase 3 (CERS3) gene in the artichoke extract of the present invention are illustrated.

[0049] Figure 9 The results confirming the anti-glycation activity of the artichoke extract of the present invention are illustrated.

[0050] Figure 10 The results confirming the pore improvement rate after using the serum formulation containing the artichoke extract of the present invention are illustrated.

[0051] Figure 11 The results confirming the reduction in sebum secretion after using an essence formulation containing the artichoke extract of the present invention are illustrated.

[0052] Figure 12 The results confirming the wrinkle improvement rate after using a face cream formulation containing the artichoke extract of the present invention are illustrated. Detailed Implementation

[0053] The preferred embodiments of the present invention will now be described in detail. However, the present invention is not limited to the embodiments described herein, but can be implemented in other forms. These embodiments are merely intended to make the description more thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

[0054] <Example 1. Preparation of Artichoke Extract by Low-Temperature Vacuum Drying>

[0055] Freshly harvested artichoke stamens were subjected to low-temperature vacuum drying at 15°C for 24 hours under a vacuum of -750 to -760 mmHg. When subjected to low-temperature vacuum drying at 10 to 20°C for 24 to 48 hours, the physical properties of the extract did not change significantly.

[0056] Next, using artichoke feedstock that had been vacuum-dried at low temperature as a baseline, 20 times its weight of a 30% (v / v) 1,3-butanediol aqueous solution was added as the extraction solvent, and extraction was performed at 60°C for 48 hours. After extraction, the extract was separated using filter paper. Finally, final filtration was performed sequentially using membrane filters with pore sizes of 0.45 μm and 0.22 μm.

[0057] For use in experiments, the extract may be concentrated or powdered, or its concentration may be adjusted to match that of other extracts after the content of the solid component has been confirmed by spectrophotometry.

[0058] <Comparative Example 1. Manufacturing of Artichoke Extract via Hot Air Drying>

[0059] Freshly harvested artichoke stamens were hot-air dried at 60°C for 24 hours. Next, the hot-air dried artichoke raw material was used to produce an extract by adding an extraction solvent in the same manner as in Example 1.

[0060] <Comparative Example 2. Manufacturing of Artichoke Extract After High Temperature and High Pressure Processing and Hot Air Drying>

[0061] Freshly harvested artichoke stamens were subjected to high-temperature and high-pressure treatment at 150°C and 20 MPa (approximately 150,012 mmHg) for 20 minutes. After the high-temperature and high-pressure treated artichoke raw material was dried with hot air at 60°C for 24 hours, an extraction solvent was added in the same manner as in Example 1 to produce an extract.

[0062] <Comparative Example 3. Preparation of Artichoke Extract After Freeze-Drying>

[0063] Freshly harvested artichoke stamens were freeze-dried at -70°C and 0.1 mmHg for 48 hours. The freeze-dried artichoke raw material was then used to produce an extract by adding an extraction solvent in the same manner as in Example 1.

[0064] <Comparative Example 4. Preparation of Artichoke Extract After Room Temperature Drying>

[0065] The freshly harvested artichoke stamens were dried in a cool, shaded place at room temperature (25°C) for 96 hours. The extract was then prepared by adding an extraction solvent to the artichoke raw material that had been dried at room temperature, following the same method as in Example 1.

[0066] <Examples 2 and 3, Comparative Examples 5 to 10>

[0067] By altering the conditions shown in Table 1 below, additional comparative extracts were produced. Furthermore, to facilitate easy comparison of the conditions of Example 1 and Comparative Examples 1 to 4, they are described in Table 1.

[0068] Table 1

[0069]

[0070]

[0071] The ethanol (99.9%) solvent and the 70% (v / v) ethanol aqueous solution solvent in Table 1 are used as representative of various alcohol solvents. Since the extraction yields of methanol (99.9%) or 70% (v / v) methanol aqueous solution solvents are similar, and the extract is to be used for cosmetic purposes, the methanol solvent is only confirmed in the preliminary test steps, and only ethanol solvents are used in the test.

[0072] <Experimental Example 1. Analysis of Active Components Using High Performance Liquid Chromatography (HPLC)>

[0073] The artichoke extracts from Examples 1 to 3 and Comparative Examples 1 to 10 were analyzed by high-performance liquid chromatography (HPLC) under the conditions shown in Table 2. The representative results of the gas chromatography confirmed by the results are shown in Figure 1, and the quantitative values ​​of the effective compounds in each extract are shown in Table 3.

[0074] In Table 3 below, "-" indicates that the corresponding compound was not detected.

[0075] Table 2

[0076]

[0077]

[0078] Table 3

[0079]

[0080] As confirmed by Figure 1 and Table 3, the extracts of Examples 1 to 3 contained approximately 3.2 to 3.5% by weight of 1,5-dicaffeoylquinic acid and chlorogenic acid. The extracts of Comparative Examples 1 and 2 contained only trace amounts or no of these compounds, while the extract of Comparative Example 3 contained approximately 2.7% by weight of each compound. The extract of Comparative Example 4 contained even lower amounts of the active compounds compared to the extract of Comparative Example 3.

[0081] Furthermore, the results of Comparative Examples 4 to 10 also confirmed that the distribution of 1,5-dicaffeoylquinic acid and chlorogenic acid can vary depending on the drying conditions or solvent extraction conditions of the artichoke.

[0082] <Experimental Example 2. Confirmation of Gene Expression Related to Skin Elasticity>

[0083] To confirm the efficacy on skin elasticity, the expression of skin elasticity-related genes in the extracts of Examples 1 to 3 and Comparative Examples 1 to 4 was evaluated.

[0084] First, human fibroblast cell line CCD-1064SK cells, purchased from the American Type Culture Collection (USA), were cultured at 5 × 10⁻⁶ cells per cell line. 5 Cells were seeded at a density of 100 cells / well in 6-well plates and cultured for 24 hours. The culture medium used for the cells was Iscove's Modified Dulbecco's Media (IMDM, Korea) containing fetal bovine serum (FBS, Gibco, USA). Next, each well was replaced with serum-free medium and treated with extracts from the samples and ascorbic acid (TCI, Japan) as a positive control. After 24 hours of treatment with each sample, the expression of skin elasticity-related genes (type I collagen (COL1A1), type III collagen (COL3A1), and elastin) was observed.

[0085] The method for extracting ribonucleic acid (RNA) from CCD-1064SK cells is described below. Cells treated with extracts from various samples were washed with phosphate-buffered saline (PBS, Samchun, Korea), and the cultured cells were then... Cell lysis was performed using (QIAGEN, USA). Next, ribonucleic acid (RNA) was isolated following the procedures provided by the manufacturer, QIAGEN. The isolated RNA was then analyzed using a ribonucleic acid (RNA) quantification kit. After quantification using a fluorometer with an RNABR Assay kit (Invitrogen, USA), complementary deoxyribonucleic acid (cDNA) was synthesized and real-time polymerase chain reaction (Real-time PCR) was performed. The cDNA synthesis was performed using a qPCRBIO cDNA Synthesis Kit (Pcrbiosystems, UK). Real-time PCR amplification was performed using a Real-time PCR kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, Pcrbiosystems UK), and the amplified products were analyzed. Forty cycles of Real-time PCR were performed at 95°C for 5 seconds and 60°C for 30 seconds. All experiments using the kit were performed according to the manufacturer's instructions. The expression levels of type I collagen (COL1A1), type III collagen (COL3A1), and elastin genes are presented proportionally based on the gene expression levels of the control group.

[0086] Table 4

[0087]

[0088]

[0089] The results are shown in Table 4. Figures 2 to 4 As shown, it can be confirmed that the expression of genes related to skin elasticity was increased in the extracts of Examples 1 to 3. These results are superior to the positive control group, ascorbic acid.

[0090] <Experimental Example 3. Evaluation of Gene Expression Related to Skin Barrier Strengthening>

[0091] To confirm the skin barrier strengthening efficacy, the skin barrier strengthening efficacy was evaluated using extracts from Examples 1 to 3 and Comparative Examples 1 to 4.

[0092] First, the human keratinocyte cell line HaCaT cells, purchased from the German cell line service company CLS (Cell Lines Service, Germany), were used at a concentration of 5 × 10⁻⁶ cells / mL. 5 Cells were seeded at a density of 100 cells / well in 6-well plates and cultured for 24 hours. The culture medium used for the cells was Dulbecco Modified Eagle Medium (DMEM, Korea) containing fetal bovine serum (FBS, Gibco, USA). Next, each well was replaced with serum-free medium and starved for 24 hours. Following this, each well was treated with the aforementioned extracts and a positive control, calcium chloride. After 24 hours of treatment with each sample, the expression of skin barrier-related genes (hyaluronic acid synthase 3 (HAS3), caspase 14 (CAS14), filaggrin (FLG), and ceramide synthase 3 (CERS3)) was observed.

[0093] The method for extracting ribonucleic acid (RNA) from HaCaT cells is described below. Cells treated with various extract samples and calcium chloride were washed with phosphate-buffered saline (PBS, Samchun, Korea). The cultured cells were then... Cell lysis was performed using (QIAGEN, USA). Next, ribonucleic acid (RNA) was isolated following the procedures provided by the manufacturer, QIAGEN. The isolated RNA was then analyzed using a ribonucleic acid (RNA) quantification kit. After quantification using a fluorometer with an RNA BR Assay kit (Invitrogen, USA), complementary deoxyribonucleic acid (cDNA) was synthesized and real-time polymerase chain reaction (PCR) was performed. The cDNA synthesis was performed using a qPCRBIO cDNA Synthesis Kit (Pcrbiosystems, UK). The amplified products were analyzed after gene amplification using a real-time polymerase chain reaction kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, Pcrbiosystems UK). Forty cycles of PCR were performed at 95°C for 5 seconds and 60°C for 30 seconds. All experiments using the kit were performed according to the manufacturer's instructions. Gene expression levels are presented relative to the control group.

[0094] Table 5

[0095]

[0096] The results are shown in Table 5. Figures 5 to 8 As shown, it can be confirmed that the expression of genes related to the skin barrier was increased in the extract treatment groups of Examples 1 to 3.

[0097] <Experimental Example 4: Evaluation of Anti-glycation Activity>

[0098] To confirm the anti-glycation effect of the extracts from Examples 1 to 3 and Comparative Examples 1 to 10, the glycation inhibition activity was measured using L-arginine and glucose.

[0099] First, 1M L-arginine and 1M glucose were dissolved in 1M phosphate buffer (pH 7.4). Then, each extract was processed to 50 to 150 μg / ml using 1M phosphate buffer.

[0100] After mixing 1M L-arginine and 1M phosphate buffer at a ratio of 1:4, the mixture was aliquoted into 96-well plates in 80 μL increments. 100 μL of each sample and 0.01M aminoguanidine (positive control) were added and mixed thoroughly. Next, glucose was diluted to a final concentration of 0.1M using 1M phosphate buffer, and the reaction was carried out at 70°C for 4 hours. The absorbance of the 96-well plate at 420 nm was measured using a spectrophotometer to confirm the degree of glycosylation.

[0101] The glycosylation test group was induced by adding 1M L-arginine and 1M glucose. In order to measure the absorbance of the sample itself, only 1M L-arginine and the sample were added without adding glucose, and the absorbance was measured.

[0102] The glycosylation inhibition rate (%) was calculated using the following formula [Formula 1], and the average value was obtained by performing three experiments. The results are shown in Table 6 below. Figure 9 As shown.

[0103] [Formula 1]

[0104] Glycosylation inhibition rate (%) = {(absorbance of the sample after reaction - absorbance of the sample before reaction) / absorbance of the glycosylation test group} × 100 [Table 6]

[0105]

[0106]

[0107] See Table 6 and Figure 9 The experimental results showed that the extracts of Examples 1 to 3 exhibited glycosylation inhibition rates of over 40% at 100 and 150 μg / ml.

[0108] The results described above confirm that artichoke extract processed by low-temperature vacuum drying can be used more effectively for anti-glycation cosmetic applications.

[0109] <Example 1 of Dosage Form and Comparative Dosage Form 1. Manufacturing of Test Products for Pore and Sebum Improvement - Serum>

[0110] To conduct subsequent tests on pore and sebum improvement, an essence formulation containing 3% by weight of the artichoke extract from Example 1 was manufactured, while purified water was added to the control group instead of the extract.

[0111] As the artichoke extract, the concentration of the extract in the essence was achieved by dissolving it in a 30% (v / v) aqueous solution of 1,3-butanediol, resulting in an extract concentration of approximately 150 μg / ml.

[0112] The specific ingredients of the serum formulation are shown in Table 7 below. Specifically, butylene glycol, propylene glycol, glycerin, disodium EDTA, carbomer, and purified water were homogenized at 1500 rpm for 3 minutes at a temperature of 45°C to 50°C. Triethanolamine was added and then stirred for 3 minutes to neutralize it. Next, 1,2-hexanediol and ethylhexylglycerin were added and stirred for 3 minutes, followed by cooling to 30°C. Then, the extract from Example 1 was added to 3% by weight and stirred and defoamed for 3 minutes to produce the serum formulation.

[0113] Table 7

[0114] raw material Dosage form example 1 (by weight %) Comparative dosage form example 1 (by weight %) Extract solution of Example 1 3.00 0 Butylene glycol 4.00 4.00 Propylene glycol 4.00 4.00 glycerin 3.00 3.00 1,2-Hexanediol 2.00 2.00 Carbomer 0.20 0.20 Triethanolamine 0.16 0.16 Ethylhexylglycerin 0.05 0.05 Disodium ethylenediaminetetraacetate 0.02 0.02 purified water margin margin total 100.00 100.00

[0115] <Example 5. Confirmation of the pore and sebum-improving effects of a serum containing artichoke extract that has undergone low-temperature vacuum drying>

[0116] Experimental Example 5-1: Confirmation of Pore-Improving Efficacy

[0117] After using the serum test product of dosage form 1 or comparative dosage form 1 daily for 4 weeks, the size and number of pores were measured using a skin diagnostic instrument (ASW-100, Aramhuvis).

[0118] The skin diagnostic instrument (ASW-100, Aramhuvis) is a diagnostic system designed to measure the condition of hair, pores, sebum, and wrinkles. Results are scored from 1 to 100, with higher scores indicating more relevant factors. In this experimental case, the score measured before use was used as a baseline, and the changes were calculated as an improvement rate, or reduction rate.

[0119] Table 8

[0120] Pore ​​score (ASW-100 measurement value) Dosage Form Example 1 Comparative Dosage Form Example 1 Improvement rate (%) 31.5 7.7

[0121] As shown in Table 8 and Figure 10 As shown, compared with before using the test product, there was an average improvement rate of 31.5% after 4 weeks of use, which confirms that the pore size was significantly reduced.

[0122] Experimental Example 5-2. Confirmation of Sebum Regulation Efficacy

[0123] After 4 weeks of daily use of the serum test product in dosage form 1 or comparative dosage form 1, sebum secretion was measured using the ASW-100 (Aramhuvis) and expressed as the rate of reduction in sebum secretion after 4 weeks, compared to the sebum secretion level before use. The results are shown in Table 9 below. Figure 11As shown.

[0124] Table 9

[0125] Sebum score (ASW-100 measurement) Dosage Form Example 1 Comparative Dosage Form Example 1 Improvement rate (%) 28.0 5.3

[0126] As shown in Table 9 and Figure 11 As shown, compared with before using the test product, after 4 weeks of use, the group using dosage form 1 showed an average improvement rate of 28%, which confirms that sebum secretion was significantly reduced.

[0127] <Example 2 of Dosage Form and Comparative Dosage Form 2. Manufacturing of a Test Product for Wrinkle Improvement - Face Cream>

[0128] Next, a face cream formulation for testing its wrinkle-improving efficacy was prepared using the same raw material extract used in Formulation Example 1. Purified water was added in place of the extract in the control group. The specific ingredients of the face cream formulation are shown in Table 10 below.

[0129] Specifically, an oil phase was prepared by melting glyceryl stearate, synthetic beeswax, arachidonic acid, behenol, arachidonic acid glucoside, cetyl ethylhexanoate, and caprylic / capric triglycerides at 75°C to 80°C. An aqueous phase was prepared by dispersing disodium EDTA, cetearyl oleate, sorbitan oleate, glycerol, propylene glycol, and carbomer in purified water. The molten oil phase was added to the aqueous phase heated to 70°C to 75°C, followed by a first emulsification at 3500 to 5000 rpm for 5 minutes using a homogenizer. Triethanolamine was added at 60°C to 65°C, followed by neutralization by stirring at 3000 to 3500 rpm for 3 minutes using a homogenizer. A second emulsification was then completed by adding sodium polymethacryloyl dimethyl taurate and stirring at 3000 to 3500 rpm for 3 minutes. 1,2-hexanediol and ethylhexylglycerin were added at 45°C and stirred for 3 minutes before cooling to 30°C. Next, the extract from Example 1 was added and stirred and defoamed for 3 minutes to produce a cream formulation.

[0130] Table 10

[0131]

[0132]

[0133] <Example 6. Confirmation of the wrinkle-improving efficacy of a face cream containing artichoke extract that has undergone low-temperature vacuum drying>

[0134] After four weeks of daily use of the face cream test product of dosage form Example 2 or comparative dosage form Example 2, the depth of wrinkles was measured using ASW-100 (Aramhuvis), and the results are shown in Table 11 below as improvement rates.

[0135] Table 11

[0136] Wrinkle score (ASW-100 measurement value) Dosage Form Example 2 Comparative Dosage Form Example 2 Improvement rate (%) 18.1 -2.2

[0137] The results are shown in Table 11 and Figure 12 As shown, compared with before using the test product, the wrinkle depth of the face cream group of dosage form 2 was significantly reduced after 4 weeks of use. The numerical results showed an average wrinkle improvement rate of 18.1%, and it can be confirmed that the face cream group of dosage form 2 showed almost no change in skin wrinkles.

[0138] The results described above confirm that cosmetic compositions containing artichoke extracts processed by low-temperature vacuum drying according to the present invention have excellent efficacy in improving skin elasticity, wrinkles, pores, and sebum regulation.

[0139] *The primer sequences used in the experiments of this invention are shown below.

[0140] Table 12

[0141]

[0142]

Claims

1. A cosmetic composition for anti-glycation, characterized in that: Artichoke (Cynara scolymus) extract containing enhanced 1,5-dicaffeoylquinic acid and chlorogenic acid.

2. The cosmetic composition for anti-glycation according to claim 1, characterized in that: The artichokes were dried under low-temperature vacuum conditions of 4 to 37°C and 0 to -760 mmHg.

3. The cosmetic composition for anti-glycation according to claim 1, characterized in that: The artichoke extract has the effects of improving pores and regulating sebum.

4. The anti-glycation cosmetic composition according to claim 1, characterized in that: The artichoke extract has skin barrier improving effects.

5. The cosmetic composition for anti-glycation according to claim 1, characterized in that: The artichoke extract is said to enhance the gene expression of hyaluronic acid synthase 3 (HAS3), caspase 14 (CAS14), filaggrin (FLG), or ceramide synthase 3 (CERS3).

6. The anti-glycation cosmetic composition according to claim 1, characterized in that: The artichoke extract has the effect of improving wrinkles or elasticity.

7. The anti-glycation cosmetic composition according to claim 1, characterized in that: It has the effect of enhancing the gene expression of type I collagen (COL 1A 1), type III collagen (COL3A1), or elastin.

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