Skin-mproving cosmetic composition comprising andrographolide and extract of bidens pilosa l.

Abstract

The present invention relates to a cosmetic composition comprising andrographolide and an extract of Bidens pilosa L. as active ingredients. The cosmetic composition of the present invention exhibits excellent effects superior to those of when each of andrographolide or an extract of Bidens pilosa L. is used alone on wrinkle alleviation, skin irritation alleviation, elasticity improvement, inflammation, and antioxidation by using andrographolide and the extract of Bidens pilosa L. together.

Description

Cosmetic composition for skin improvement comprising andrographolide and Bidens pilosa extract

[0001] The present invention relates to a cosmetic composition for skin improvement comprising andrographolide and Bidens pilosa extract as active ingredients, and a method for preparing the same.

[0002] Skin aging includes natural aging caused by genetic factors and external aging caused by the living environment. The biggest external cause of skin aging is ultraviolet (UV) radiation. UV rays penetrate the skin, inducing inflammatory and oxidative reactions that cause skin damage, and also directly affect DNA, disrupting skin homeostasis. UV rays promote the expression of Matrix Metalloproteinase-1 (MMP-1), an enzyme that breaks down Collagen Type 1 and contributes to the formation of skin wrinkles.

[0003] Meanwhile, retinol is a type of Vitamin A known for its excellent anti-aging effects and is widely used in anti-wrinkle cosmetics. Retinol is known to act on skin fibroblasts to inhibit MMP-1 enzyme expression and increase collagen type-1 synthesis, thereby improving skin wrinkles. However, retinol has the disadvantage of being difficult to use during the day due to its susceptibility to skin irritation and photodegradation.

[0004] To solve these problems, there is a need to develop effective materials that exhibit excellent skin improvement like retinol while being non-irritating.

[0005] Accordingly, the inventors have made diligent research efforts to develop an effective material that exhibits excellent skin improvement effects without irritation.

[0006] As a result, the present invention was completed by preparing a cosmetic composition containing andrographolide and Bidens pilosa extract as active ingredients.

[0007] Accordingly, the objective of the present invention is to provide a cosmetic composition comprising andrographolide and Bidens pilosa L. extract as active ingredients.

[0008] Another objective of the present invention is to provide a cosmetic composition for improving skin comprising andrographolide and Bidens pilosa L. extract as active ingredients.

[0009] Another objective of the present invention is to provide skin-improving uses for andrographolide and Bidens pilosa L. extracts.

[0010] Another objective of the present invention is to provide a skin improvement method comprising the step of applying a cosmetic composition containing an effective amount of andrographolide and Bidens pilosa L. extract to the skin of a subject in need thereof.

[0011] The present invention will be described in detail below.

[0012] In the present invention, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may additionally include other components.

[0013] One aspect of the present invention is a cosmetic composition comprising andrographolide and Bidens pilosa L. extract as active ingredients.

[0014] In the present invention, andrographolide is a diterpenoid-based substance extracted from plants. It is a material extracted from the stems or leaves of *Andrographis paniculata* and possesses anticancer and anti-inflammatory effects. Andrographolide may have a structure such as the following chemical formula I.

[0015] [Chemical Formula I]

[0016]

[0017] In the present invention, the method of obtaining andrographolide is not particularly limited and may be obtained from nature, synthesized by methods known in the art, or commercially available material. Furthermore, the andrographolide may include all derivatives within a range predictable by those skilled in the art, and may be included without limitation as long as they have the same effect in the present invention.

[0018] In the present invention, “Bidens Pilosa L.” is an annual herb belonging to the genus Bidens and the family Compositae. Bidens Pilosa has been used for centuries in traditional medicine and as food in many parts of the world, and Bidens Pilosa extract has been traditionally used in Africa to treat malaria.

[0019] In the present invention, the extract of *Devil's Needle* may mean that it is extracted from one or more selected from the group consisting of the leaves, stems, roots, and whole plant of the *Devil's Needle* plant, and preferably, it may mean that it is extracted from the whole plant of the *Devil's Needle* plant, but is not limited thereto.

[0020] According to an embodiment of the present invention, when skin fibroblasts were treated with both andrgrafoliide and Bidens pilosa extract, the expression of the RXRA (Retinoid X receptor alpha) gene was promoted by more than 1.2 times compared to the untreated group (Example 1). In addition, when skin fibroblasts were treated with both andrgrafoliide and Bidens pilosa extract, collagen synthesis increased compared to the untreated group, and collagen synthesis increased statistically significantly (P<0.05) when treated at ratios of 1:3, 1:5, and 1:10 (Example 2-1). Furthermore, when skin fibroblasts were treated with a mixture of andrgrafoliide and Bidens pilosa extract at different concentrations, MMP-1 synthesis decreased (Example 2-2) and Fibrillin-1 increased (Example 2-3) compared to treatment with each extract alone. In addition, when skin keratinocytes (HaCaT) were simultaneously treated with andrgrapolid and Bidens pilosa extract, IL-1a, which increases due to UV, decreased compared to when treated alone (Example 3-1). In addition, when skin fibroblasts were simultaneously treated with andrgrapolid and Bidens pilosa extract, the cell viability, which decreases due to hydrogen peroxide, increased compared to when treated alone (Example 3-2).

[0021] Through this, it was confirmed that the andrographolide and Ulsan Dokkaebi extract of the present invention are highly effective for skin improvement, such as wrinkle reduction, skin irritation reduction, elasticity improvement, inflammation relief, and antioxidant effects.

[0022] As used herein, the term “collagen” refers to a fibrous protein that constitutes the majority of various tissues in the body, such as skin and cartilage, and is produced and secreted by skin fibroblasts, which are common cells in the skin. Collagen is reduced by UV radiation, and this reduction in collagen decreases skin elasticity and promotes wrinkle formation. Furthermore, UV radiation promotes the expression of MMP-1, one of the collagen-degrading enzymes, thereby accelerating collagen degradation.

[0023] The term “Fibrillin-1” as used herein refers to a protein distributed in the extracellular matrix that is important for maintaining tissue structure. Fibrillin-1 is also known to be distributed within the skin and plays an important role in maintaining skin elasticity. Fibrillin-1 is reduced by UV radiation.

[0024] As used herein, the term “improvement of skin irritation” means alleviating and soothing erythema, itching, pain, etc., associated with mild irritation, and may be a concept that includes the alleviation of skin irritation. Examples include, but are not limited to, alleviation of skin itching, alleviation of skin pain, reduction of skin moisture loss and / or reduction of redness.

[0025] As used herein, the term “inflammation” refers to a defense mechanism designed to protect tissues from damage or infection from outside or inside. The purpose of inflammation is to remove substances that cause damage, but an excessive inflammatory response leads to tissue damage. In the present invention, the improvement, soothing, and regeneration of skin irritation may be achieved by inhibiting inflammatory substances (IL-1a), but are not limited thereto.

[0026] The term “antioxidant” in this specification means inhibiting oxidative stress caused by hydrogen peroxide. When oxidative stress occurs, cell death takes place, and inhibiting such cell death is also an antioxidant action.

[0027] In this specification, the term “extract” refers to a product such as a liquid component obtained by immersing a target substance in various solvents and then extracting it for a certain period of time at room temperature or under heated conditions, or a solid component obtained by removing the solvent from said liquid component. The extraction may be performed one or more times, but is not limited thereto. Furthermore, it may be comprehensively interpreted to include all of the filtered liquid, diluted liquid, concentrates thereof, modified products thereof, purified products, dried products thereof, etc. of said product. Accordingly, the Ulsan Devil’s Needle extract provided in the present invention may be interpreted to include the extract itself, extracts of all formulations that can be formed using said extract, and / or dried products thereof, such as an extract obtained by extracting Ulsan Devil’s Needle, a purified product obtained by enzymatic hydrolysis of said extract, or a mixture thereof.

[0028] In addition, the method for obtaining the above extract is not limited thereto, as long as it can yield an extract that exhibits skin-improving effects, for example, wrinkle improvement, skin elasticity enhancement, anti-inflammatory or antioxidant effects.

[0029] In the present invention, the extract of *Devil's Needle* may be extracted by one or more methods selected from the group consisting of supercritical extraction, hot water extraction, cold maceration extraction, ultrasonic extraction, and reflux cooling extraction, and preferably may be extracted by a supercritical CO2 extraction method, but is not limited thereto.

[0030] In the present invention, andrographolide may be included in an amount of 0.0001 to 10 weight%, for example, 0.01 to 1 weight%, relative to the weight of the total composition, but is not limited thereto.

[0031] In the present invention, the Ulsan Dokkaebi extract may be included in an amount of 0.0001 to 10 weight%, for example, 0.1 to 2 weight%, relative to the weight of the total composition, but is not limited thereto.

[0032] In the present invention, the cosmetic composition may contain andrographolide and Bidens pilosa extract in a ratio of 1:2 to 1:12, for example, 1:3 to 1:10, but is not limited thereto.

[0033] The ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the active ingredients as active ingredients, such as, for example, antioxidants, stabilizers, solubilizers, vitamins, UV absorbers, preservatives, pH adjusters, colorants, pigments, and fragrances, and carriers.

[0034] The cosmetic composition of the present invention may be prepared in any formulation conventionally manufactured in the art, for example, as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream, and spray, but is not limited thereto. More specifically, it may be prepared in the form of a skin toner, lotion, softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder.

[0035] In the present invention, the andrographolide and Bidens pilosa extract in the cosmetic composition may be loaded into liposomes, but are not limited thereto.

[0036] In this specification, the term “liposome” refers to a colloidal particle formed by the mutual spontaneous bonding and alignment of particles composed of amphiphilic molecules having a water-soluble head and an insoluble tail, having a double membrane structure similar to a cell membrane, and compositions containing such a liposome can be expected to have effects such as restoring damaged skin barriers, strengthening skin barriers, and alleviating skin irritation. In addition, the liposome has the effect of stably maintaining the efficacy of actin, actigenin, or mixtures thereof for a long period of time. In the present invention, the liposome refers to a liposome having the form of a conventional liposome, with an average particle diameter of 1 to 100 nm.

[0037] In the present invention, liposomes can be prepared by a mixture comprising an oily component, a phospholipid, a surfactant, a polyol, and water.

[0038] In the above, the oil component may be any of the various oils known in the art. Preferably, hydrocarbon oils such as hexadecane and paraffin oil; ester-based synthetic oils; silicone oils such as dimethicone and cyclomethicone; animal and vegetable oils such as sunflower oil, corn oil, soybean oil, avocado oil, sesame oil, and fish oil; ethoxylated alkyl ether oils; propoxylated alkyl ether oils; sphingonoid lipids such as phytosphingosine, sphingosine, and sphinganine; cerebroside; cholesterol; sitosterol; cholesteryl sulfate; sitosteryl sulfate; C10-40 fatty alcohols and mixtures thereof may be used. More preferably, sitosterol may be used.

[0039] The phospholipids used in the preparation of the liposomes of the present invention and in the examples and comparative examples are used as amphoteric lipids and may include natural phospholipids such as egg yolk lecithin, soybean lecithin, or sphingomyelin, and / or synthetic phospholipids such as hydrogenated lecithin, hydrogenated phosphatidylcholine, dipalmitoyl phosphatidylcholine, or hydrogenated lecithin.

[0040] Any surfactant known in the art may be used for the preparation of the liposomes of the present invention and for the examples and comparative examples. For example, anionic surfactants, cationic surfactants, amphoteric surfactants, or nonionic surfactants may be used, and preferably, nonionic surfactants may be used. Specific examples of nonionic surfactants may include sorbitan stearate, sorbitan monostearate, sorbitan sesquioleate, polysorbate 20, glyceryl monostearate, polyglyceryl-10 laurate, PEG-100 stearate, alkoxylated alkyl ether, alkoxylated alkyl ester, alkyl polyglycoside, polyglyceryl ester, or sugar ester.

[0041] The polyols mentioned above are not particularly limited, but may preferably be selected from the group consisting of glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, methylpropanediol, isopropylene glycol, pantylene glycol, erythritol, xylitol, sorbitol, and mixtures thereof.

[0042] Another aspect of the present invention is a cosmetic composition for improving skin comprising andrographolide and Bidens pilosa L. extract as active ingredients.

[0043] In the present invention, skin improvement may be wrinkle improvement, skin irritation improvement, skin elasticity enhancement, anti-inflammatory, or antioxidant, but is not limited thereto.

[0044] Another aspect of the present invention is the use of andrographolide and Bidens pilosa L. extracts for skin improvement.

[0045] Another aspect of the present invention provides a skin improvement method comprising the step of applying a cosmetic composition containing an effective amount of andrographolide and Bidens pilosa L. extract to the skin of a subject in need thereof.

[0046] The present invention relates to a cosmetic composition comprising andrographolide and Bidens pilosa extract as active ingredients. The cosmetic composition of the present invention, when using andrographolide and Bidens pilosa extract together, exhibited superior effects in improving wrinkles, skin irritation, elasticity, inflammation, and antioxidant properties compared to when each was used alone.

[0047] Figure 1 is a graph showing the comparison of the expression levels of the RXRA (Retinoid X receptor alpha) gene after treatment with andrographolide and Ulsan devil's needle extract.

[0048] Figure 2 is a graph showing the collagen protein secretion amount (%) measured and compared after treatment with andrographolide and Ulsan devil's needle extract.

[0049] Figure 3 is a graph showing the comparison of MMP-1 protein secretion (%) measured after treatment with andrographolide and Ulsan devil's needle extract.

[0050] Figure 4 is a figure showing the comparison of fibrillin-1 protein expression after treatment with andrographolide and Ulsan devil's needle extract.

[0051] Figure 5 is a graph showing the area (%) of fibrillin-1 protein measured and compared after treatment with andrographolide and Devil's Needle extract.

[0052] Figure 6 is a graph showing the IL-1a protein secretion amount (%) measured and compared after treatment with andrographolide and Ulsan devil's needle extract.

[0053] Figure 7 is a graph showing the cell viability (%) reduced by hydrogen peroxide after treatment with andrographolide and Bidens pilosa extract.

[0054] The advantages and features of the present invention and the methods for achieving them will become clear by referring to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below but may be implemented in various different forms; these embodiments are provided merely to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention, and the present invention is defined only by the scope of the claims. The raw materials used in the present invention were purchased from raw material suppliers that are typically available for purchase.

[0055]

[0056] Preparation Example 1. Preparation of andrographolide and Bidens pilosa extract

[0057] I purchased and used both Ulsan devil's needle extract (Revinage®, Chemyunion, Brazil) and andrographolide (AlphaTriptec).

[0058]

[0059] Preparation Example 2. Liposomalization of andrographolide and Bidens pilosa extract

[0060] Dipropylene glycol, hydrogenated lecithin, cholesterol, stearic acid, sorbitan stearate, andrographolide, and Bidens pilosa extract were mixed and dissolved at 350 rpm at 80 ℃ as shown in the ingredients of Table 1 below. Then, purified water at 80 ℃ was added and mixed at 350 rpm for 30 minutes, followed by natural cooling at room temperature to prepare liposomes containing andrographolide and Bidens pilosa extract.

[0061] Ingredient Content (Unit: Weight%) Dipropylene Glycol 33.33 Hydrogenated Lecithin 2 Cholesterol 0.1 Stearic Acid 0.1 Sorbitan Stearate 0.1 Andrographolide 0.001 Devil's Needle Extract 0.1 Purified Water Residue

[0062]

[0063] Examples

[0064] In this example, the efficacy of andrographolide and Bidens pilosa extract in promoting retinoid X receptor alpha gene expression (Example 1), collagen synthesis (Example 2-1), MMP-1 expression reduction (Example 2-2), Fibrillin-1 expression promotion (Example 2-3), skin inflammation improvement (Example 3-1), and inflammation relief and antioxidant efficacy was confirmed.

[0065]

[0066] Example 1. Evaluation of Retinoid X Receptor Alpha Gene Expression of Andrographolide and Bidens pilosa Extract

[0067] To measure the expression level of RXRA, a retinoid X receptor gene, in the cell nucleus, human dermal fibroblasts were seeded into 6-well plates at a density of 3 x 10^5 cells / well and cultured overnight in a CO2 incubator. After 24 hours, andrographolide and Bidens pilosa extracts were mixed into DMEM media (Dulbecco Modified Eagle Medium) at the concentrations shown in Table 1 below and then treated with the cells. After 24 hours, RNA was extracted according to the protocol using a Total RNA Extraction Kit (Bioneer, Korea), and Real-time qPCR was performed to compare and analyze the expression levels of the RXRA (Retinoid X receptor alpha) gene. The measurement results are shown in Table 2 and Figure 1.

[0068] Treatment Group Applied Concentration RXR Gene Expression Colby Equation Predicted Values ​​Untreated - 1 - Andrographolide 1 ppm 1.07 - Devil's Needle Extract 10 ppm 1.03 - Andrographolide + Devil's Needle Extract - 1.21 1.10

[0069]

[0070] Experimental results showed that when 1 ppm of andrographolide and 10 ppm of Ulsan devil's needle extract were treated together, RXRA expression was promoted by more than 1.2 times compared to the untreated group (Fig. 1).

[0071]

[0072] Example 2. Evaluation of the efficacy of andrographolide and Bidens pilosa extract in improving wrinkles or elasticity

[0073] 2-1. Evaluation of Collagen Synthesis Enhancement by Andrographolide and Bidens pilosa Extract

[0074] Andrographolide and Bidens pilosa extracts were mixed into DMEM media at the concentrations shown in Table 2 below, and then treated with cells.

[0075] Specifically, to evaluate collagen synthesis ability, skin fibroblasts were first seeded into a 24-well plate at a density of 2 x 10^4 cells / well and cultured overnight in a CO2 incubator. After 24 hours, the substances were mixed into DMEM media (Dulbecco Modified Eagle Medium) at the concentrations shown in Table 2 below and then treated with the cells. Next, the culture medium was collected, and the procollagen secreted into the culture medium was measured using a Human Pro-Collagen I alpha 1 DuoSet ELISA kit (DY6220-05, R&D systems, USA), and the results are shown in Table 3 and Figure 2.

[0076] Treatment Group | Applied Concentration | Collagen Secretion Amount (%) | Colby Formula Predicted Value (%) Untreated - 100 | Andrographolide 1 ppm | 118 | Devil's Needle Extract 10 ppm | 107 | Andrographolide + Devil's Needle Extract - 132 | 123.7

[0077]

[0078] Experimental results showed that when andrgrapolid and Bidens pilosa extract were treated simultaneously, the amount of collagen synthesis increased compared to when treated alone (Fig. 2).

[0079] In addition, andrographolide and Bidens pilosa extracts were mixed in DMEM media according to the concentrations and ratios shown in Table 3 below, and then treated with cells. Cells were cultured and treated in the same manner as in the evaluation of collagen synthesis ability to obtain culture medium, and the procollagen secreted into the culture medium was measured using a Human Pro-Collagen I alpha 1 DuoSet ELISA kit (DY6220-05, R&D systems, USA). Student's t-test was used for statistical analysis to obtain the P-value.

[0080] Treatment Group Collagen Secretion Amount Colby Equation Predicted Value t-test p-value (vs 1:1) t-test p-value (vs 1:20) Untreated 100---Andrographolide 1ppm 136---Ulsan Devil's Needle Extract 1ppm 99---3ppm 110---5ppm 100---10ppm 105---20ppm 102---Combination 1:1130135.36-p=0.1517 1:3158142.4p=0.0116p=0.007 51:5145136p=0.0425p=0.0154 1:10169139.2p=0.0193p=0.0105 1:20121137.28p=0.1517-

[0081]

[0082] Experimental results confirmed that when andrographolide and Bidens pilosa extract were simultaneously treated at ratios of 1:3, 1:5, and 1:10, the amount of collagen synthesis increased compared to treatment with each individual. Additionally, it was confirmed that the amount of collagen synthesis increased statistically significantly (P<0.05) when treated at ratios of 1:3, 1:5, and 1:10 compared to treatment at ratios of 1:1 and 1:20.

[0083] 2-2. Evaluation of MMP-1 Expression of Andrographolide and Bidens pilosa Extract

[0084] Andrographolide and Bidens pilosa extracts were mixed into DMEM media at the concentrations shown in Table 5 below and then treated with cells. Specifically, to evaluate MMP-1 expression, skin fibroblasts were first seeded at 2 x 10^4 cells / well in a 24-well plate and cultured overnight in a CO2 incubator. After 24 hours, the substances were mixed into DMEM media (Dulbecco Modified Eagle Medium) at the concentrations shown in Table 5 below and then treated with the cells. Subsequently, the culture medium was collected, and MMP-1 secreted into the culture medium was measured using a Human Total MMP-1 DuoSet ELISA kit (DY901B-05, R&D systems, USA). The results are shown in Table 5 and Figure 3.

[0085] Treatment Group Applied Concentration MMP1 Secretion Amount (%) Colby Equation Predicted Value (%) Untreated - 100 - Andrographolide 1 ppm 110 - Devil's Needle Extract 10 ppm 69 - Andrographolide + Devil's Needle Extract - 58 8 2.1

[0086]

[0087] Experimental results confirmed that when andrographolide and Bidens pilosa extracts were treated simultaneously, the amount of MMP-1 synthesized decreased compared to when treated alone (Fig. 3).

[0088] 2-3. Evaluation of Fibrillin-1 Expression by Andrographolide and Bidens pilosa Extract

[0089] Andrographolide and Bidens pilosa extracts were mixed into DMEM media at the concentrations shown in Table 5 below, and then treated with cells.

[0090] Specifically, to evaluate fibrillin-1 expression, skin fibroblasts were first seeded into 12-well plates at a density of 1 x 10^5 cells / well and cultured overnight in a CO2 incubator. After 24 hours, the substances were mixed into DMEM media (Dulbecco Modified Eagle Medium) at the concentrations shown in Table 5 below and then treated with the cells. After 24 hours, cells were fixed by treating with 4% paraformaldehyde (P2031, Biosesang) for 10 minutes, followed by treatment with 0.1% Triton X-100 for 10 minutes. After washing three times with PBS (Phosphate buffered saline), the blocking process was performed using PBS containing 5% FBS. Subsequently, the cells were treated with Anti-Fibrillin 1 antibody (ab53076, Abcam) for 24 hours, followed by treatment with Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) (ab150077, Abcam) for 1 hour. To stain the cell nuclei, DAPI (4',6-diamidino 2-phenylindole) was diluted to a concentration of 5 mg / ml in PBS and treated for 10 minutes. Next, the fluorescence signal was measured using a fluorescence microscope, and the elastic protein expression was compared by quantitatively measuring the intensity of the fluorescence signal.

[0091] Treatment Group | Applied Concentration | Elasticity | Protein (%) | Colby Equation | Predicted Value (%) Untreated | Untreated - 100 | UV Untreated - 60 | Andrographolide 1 ppm | 90 | Devil's Needle Extract 10 ppm | 92 | Andrographolide + Devil's Needle Extract - 112 | 106

[0092]

[0093] Experimental results confirmed that when andrgrapolid and Bidens pilosa extracts were treated simultaneously, fibrillin-1 increased compared to when treated alone (Fig. 4).

[0094] In Example 2, it was confirmed that when andrographolide and Ulsan devil's needle extract were treated, the collagen secretion amount (%) increased, the expression of the collagen enzyme MMP-1 decreased, and the expression of fibrillin-1, a protein important for maintaining skin elasticity, increased (Fig. 5).

[0095] Through this, it was confirmed that the andrographolide and Ulsan Dokkaebi extract of the present invention have excellent effects in improving wrinkles and elasticity.

[0096]

[0097] Example 3. Evaluation of skin irritation, skin inflammation improvement, and antioxidant effects of andrographolide and Bidens pilosa extract

[0098] 3-1. Evaluation of Skin Inflammation Improvement by Andrographolide and Bidens pilosa Extract

[0099] To evaluate IL-1α expression, skin keratinocytes (HaCaT) were first seeded into 24-well plates at a density of 2*10^4 cells / well and cultured overnight in a CO2 incubator. After 24 hours, DMEM media (Dulbecco Modified Eagle Medium) was mixed with andrographolide at a concentration of 1 ppm and Bidens pilosa extract at a concentration of 10 ppm, and the cells were treated. Subsequently, the culture medium was collected, and IL-1α secreted into the culture medium was measured using a Human IL-1 alpha / IL-1F1 DuoSet ELISA kit (DY200-05, R&D systems, USA). The results are shown in Table 7 and Figure 6.

[0100] Treatment Group Applied Concentration IL-1a Secretion (%) Colby Prediction (%) Untreated Untreated -100 -UV Untreated -841 -Andrographolide 1ppm 800 -Believing Devil's Needle Extract 10ppm 744 -Andrographolide + Believing Devil's Needle Extract -596698

[0101]

[0102] Experimental results showed that when andrgrapolid and Bidens pilosa extract were treated simultaneously, IL-1a, which increases with UV, was reduced compared to when treated alone (Fig. 6).

[0103] 3-2. Evaluation of Antioxidant Activity of Andrographolide and Bidens pilosa Extract

[0104] To evaluate the antioxidant activity, skin fibroblasts were first seeded into a 96-well plate at a density of 1 x 10^4 cells / well and cultured overnight in a CO2 incubator. After 24 hours, 0.8 mM hydrogen peroxide and the substance were mixed into DMEM media (Dulbecco Modified Eagle Medium) at the concentrations shown in Table 6 below and then treated with the cells. After 24 hours, to measure cell viability, Cell Counting Kit-8 (CK04, Dojindo) was additionally applied, and after 1 hour, the optical density at a wavelength of 450 nm was measured. The results are shown in Table 8 and Figure 7 below.

[0105] Treatment Group | Applied Concentration | Cell Viability (%) | Colby Predicted Value (%) Untreated | Untreated | Untreated | -100 | -Hydrogen Peroxide H2O2 | -83 | -Andrographolide 1ppm | -94 | -Bell's Needle Extract 10ppm | -86 | -Andrographolide + Bell's Needle Extract | -97 | 96

[0106]

[0107] Experimental results showed that when andrgrapolid and Bidens pilosa extracts were treated simultaneously, the cell viability reduced by hydrogen peroxide increased compared to when treated alone (Fig. 7).

[0108] In Example 3, it was confirmed that IL-1a, a skin inflammatory factor, was reduced and cell viability, which is reduced by hydrogen peroxide, was increased.

[0109] Through this, it was confirmed that the andrographolide and Ulsan Dokkaebi extract of the present invention are excellent for alleviating inflammation and providing antioxidant effects.

Claims

1. A cosmetic composition comprising andrographolide and Bidens pilosa L. extract as active ingredients.

2. In Paragraph 1, The above-mentioned Ulsan devil's needle extract is a cosmetic composition extracted by one or more methods selected from the group consisting of supercritical extraction, hot water extraction, cold maceration extraction, ultrasonic extraction, and reflux cooling extraction.

3. In Paragraph 1, A cosmetic composition in which the above andrographolide is included in an amount of 0.0001 to 10 weight% relative to the weight of the total composition.

4. In Paragraph 1, A cosmetic composition in which the above-mentioned Ulsan devil's needle extract is included in an amount of 0.0001 to 10 weight% relative to the weight of the total composition.

5. In Paragraph 1, The above cosmetic composition comprises andrographolide and Bidens pilosa extract in a ratio of 1:2 to 1:

12.

6. In Paragraph 1, A cosmetic composition in which the andrographolide and the extract of Bidens pilosa are contained within liposomes.

7. A cosmetic composition for skin improvement comprising andrographolide and Bidens pilosa L. extract as active ingredients.

8. In Paragraph 7, A cosmetic composition for skin improvement in which the above skin improvement is wrinkle improvement, skin irritation improvement, skin elasticity enhancement, anti-inflammatory or antioxidant.

9. A skin improvement method comprising the step of applying a cosmetic composition containing an effective amount of andrographolide and Bidens pilosa L. extract to the skin of a subject in need.

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