KR102403451B1 - A multifunctional cosmetic composition comprising peptides complex and natural complex extracts - Google Patents

Abstract

The present invention relates to a multifunctional cosmetic composition containing a peptide complex and a natural complex extract as active ingredients. According to the present invention, provided is a functional cosmetic composition with less simulation and minimized skin damage by external stimulation.

Description

A multifunctional cosmetic composition containing a peptide complex and a natural complex extract

The present invention relates to a multifunctional cosmetic composition comprising a low molecular weight peptide complex and natural ingredients as active ingredients.

The skin is a membrane that covers the outside of the body and performs various physiological functions to protect the body from environmental factors such as various external stimuli, obstacles, and dryness, thereby protecting and controlling internal organs and other internal organs. do.

However, due to aging in the living body, the secretion of various hormones that control metabolism internally is decreased, and the function of immune cells and the activity of cells are reduced.

In addition, externally, the UV content in the sun's rays increases due to the destruction of the ozone layer, and the excessive physical and chemical stimulation and stress caused by the increase of free radicals and active harmful oxygen as environmental pollution intensifies is the normal function of the skin. It can damage the skin by promoting the formation of spots, freckles, etc. and aging of the skin due to deterioration, melanin deposition.

Our body performs a skin regeneration process on damaged skin. Skin regeneration is a tissue response to damage and is a process of tissue repair, such as chemotaxis, cell differentiation and replication, matrix protein synthesis, vascularization, and wound reconstitution. It is a complex biological process involving

Therefore, in order to prevent skin aging and damage caused by internal and external factors to maintain healthy and beautiful skin, physiologically active substances obtained from various animals, plants, microorganisms, etc. are added to cosmetics and used.

Efforts to maintain skin health by maintaining the inherent function of the skin and activating skin cells through various physiologically active substances as described above and promoting skin metabolism are continuing, and cosmetics and external preparations for skin are being developed accordingly.

However, these substances have limited usage due to safety issues such as irritation, erythema, and redness when applied to the skin, or have insignificant effects, making it difficult to actually expect skin function improvement effects.

Therefore, there is a demand for the development of a new composition for external application for skin that is safer and more effective than the existing composition for external application for skin.

Recently, in the cosmetic industry, a number of products using natural products have been developed to reduce skin irritation caused by chemicals.

Natural ingredients have fewer side effects on the skin, and as consumers' response to cosmetics using natural ingredients increases, the value of development as a cosmetic raw material is increasing.

Therefore, the demand for cosmetics using natural ingredients with few side effects and high consumer response is increasing, and various studies on the development of multi-functional cosmetics with excellent biosafety and skin improvement effects are being actively conducted. have.

The present invention is to solve the problems of the prior art described above, and an object of the present invention is to provide a functional cosmetic composition that has little irritation to the skin and minimizes skin damage caused by external stimuli.

According to an aspect of the present invention, a peptide complex comprising H-decapeptide-7 and H-peptapeptide-19; A natural complex extract comprising a lotus callus culture extract and a fermented callus culture extract; and raspberry ketone; as an active ingredient, a cosmetic composition is provided.

In one embodiment, the peptide complex further comprises H-octapeptide-4 and H-oligopeptide-9, and the natural complex extract further comprises a rice callus culture extract and a Myrotamnus flabellifolia callus culture extract. can do.

In one embodiment, the peptide complex further comprises acetylhexapeptide-8, and the natural complex extract may further comprise centella asiatica extract.

In one embodiment, the cosmetic composition may further include niacinamide, adenosine, allantoin, tranexamic acid, and dipotassium glycyrrhizate.

In one embodiment, the cosmetic composition further comprises alpha-bisabolol, hydrolyzed glycosaminoglycan, sodium hyaluronate, hydrolyzed hyaluronic acid, and hyaluronic acid. may include

In one embodiment, the cosmetic composition may be for skin whitening or wrinkle improvement.

In one embodiment, the cosmetic composition may be for acne improvement.

In one embodiment, the cosmetic composition may be for skin moisturizing or skin texture improvement.

The cosmetic composition according to one aspect of the present invention has less skin irritation and has a remarkably excellent skin improvement effect due to the synergistic effect of each active ingredient.

It should be understood that the effects of the present invention are not limited to the above-described effects, and include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

The terms used in this specification have been selected as currently widely used general terms as possible while considering the functions in the present invention, but these may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, and the like.

In addition, in a specific case, there is a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the corresponding invention. Therefore, the term used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, rather than the name of a simple term.

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

Numerical ranges are inclusive of the values defined in that range. Every maximum numerical limitation given throughout this specification includes all lower numerical limitations as if the lower numerical limitation were expressly written. Every minimum numerical limitation given throughout this specification includes all higher numerical limitations as if the higher numerical limitation were expressly written. Any numerical limitation given throughout this specification shall include all numerical ranges within the broader numerical range, as if the narrower numerical limitation were expressly written.

Hereinafter, examples of the present invention will be described in detail, but it is obvious that the present invention is not limited by the following examples.

One aspect of the present invention is a peptide complex comprising H-decapeptide-7 and H-peptapeptide-19; A natural complex extract comprising a lotus callus culture extract and a fermented callus culture extract; and raspberry ketone; provides a cosmetic composition comprising as an active ingredient.

The cosmetic composition contains a low molecular weight complex peptide component as an active ingredient, so that it has excellent biosafety and skin improvement activity, as well as less skin irritation.

A general rule for naming such peptides may be based on a three-letter or one-letter amino acid code, unless specifically indicated otherwise.

For example, the center of the amino acid structure is indicated by a three-letter code (eg, Ala, Lys), and a D-conformation (eg, D-Ala, D-Lys) is specified by writing “D-” in front of the three-letter code. If not indicated, it can be assumed to be an L-stereoscopic form. The amino acid residues constituting the peptide may be natural or non-natural amino acid residues.

The present inventors have identified that the peptides and natural ingredients interact to effectively inhibit skin damage and aging that may be caused by various factors, and through this, various skin improvement uses have been derived.

The peptide can be prepared according to chemical synthesis methods known in the art, in particular solid-phase synthesis techniques (Merrifield, J. Amer. Chem. 1963, Soc. 85:2149-54; Stewart, et al. Solid Phase Peptide Synthesis, 2nd. ed., Pierce Chem. 1984, Co.: Rockford, 111).

The "comprising as an active ingredient" may mean containing an effective amount that can exhibit a skin improvement effect, for example, wrinkle improvement, skin elasticity improvement, skin whitening, antioxidant effect, and the like.

The “SH-decapeptide-7” is a single-chain human peptide that is synthesized identically to a part of enkephalin, a type of protein, and consists of 10 amino acids consisting of glycine, leucine, lysine, phenylalanine, serine, threonine and tyrosine.

The “SH-peptapeptide-19” is a single-chain human peptide synthesized identically to the met-enkephalin protein consisting of five amino acids: glycine, methionine, phenylalanine, and tyrosine.

The “callus” is a tissue formed when a plant is injured, and refers to a cell mass in an initial stage cultured in a solid medium by inducing a callus.

The "cells differentiated from the callus" refer to the stage from the initial stage of induced in the callus cultured in a solid medium to the stage of differentiation into a specific organ by treatment with a plant hormone. The specific organ may be a leaf, a stem or a root.

The cells differentiated from the shingi callus may include a cell differentiated from a callus to a leaf, a cell differentiated from a callus to a stem, and a cell differentiated from a callus to a root.

Typically, the time required for differentiation into a specific organ of woody plants is about 1 to 60 days, and specifically may have a differentiation period of 1 to 45 days.

The “lotus flower ( Nelumbo nucifera )” is a perennial plant planted in ponds and reservoirs, and is native to southern Asia and northern Australia. The main components of the lotus are d-(-)-N-norarmefabine, liriodenine, N-nornuciferin, and nuciferin in the calyx, and flavonoids luteolin-7-glucoside, quercetin- There are 7-glucoside and kaempferol-3-glucosylglucoside (neluboside), and it is known to be used as an astringent, hemostasis, and tonic for oil wells, blood clots, fistula, hematemesis, and nervous breakdown.

The above “ Pungran (Neofinetia falcate Hu )” is a native Korean orchid that grows wild in the southern coastal area, Jeju Island, the archipelago island, and Hongdo Island on the west coast. The root of the fern is a thick string, and it shows the strong vitality of the epiphyte that sucks nutrients and moisture from rocks or trees.

The raspberry ketone is a major aromatic compound of red raspberry and is known as a component that regulates adiponectin, a protein used by the body to control metabolism.

The peptide complex further comprises H-octapeptide-4 and H-oligopeptide-9, and the natural complex extract may further include a rice callus culture extract and a Myrotamnus Flabellifolia callus culture extract.

The “SH-octapeptide-4” is a single-chain human peptide that is synthesized identically to a part of enkephalin, a type of protein, and consists of 8 amino acids consisting of glycine, histidine, leucine, lysine, phenylalanine and tyrosine.

The “SH-oligopeptide-9” is a single-chain synthetic human peptide obtained by fermenting E. coli and contains 11 amino acids.

The "rice ( Oryzasativa L. )" is an annual herb, and is divided into rice, paddy rice, and sticky rice according to the viscosity of rice grains. According to botanical taxonomy, rice belongs to the angiosperm plant in which the ovule (the organ that becomes a seed after it is formed in the ovary) among seed plants is surrounded by the ovary.

The rice contains about 75% of starch, about 8% of gray matter, 0.5 to 1% of fat, and a small amount of B vitamins. The fat includes ester-type cholesterol and free cholesterol, brassicasterol, stigmasterol, sitosterol, monoglyceride, diglyceride, triglyceride, phospholipid, Contains free fatty acids.

The " Myrothamnus flabellifolia " is a shrub that has a unique strategy to adapt to harsh environments as a plant that grows mainly in the African wilderness. The Myrotamnus flabellifolia is also nicknamed as an Easter plant because of its characteristic ability to sprout green leaves and bloom in a short time even with very little rain after a long drought. The Myrotamnus flabellifolia is traditionally used for coughing, cold (tea form), wounds, burns, abrasions (leaf parts), and skin irritation improvement (ointment form).

The peptide complex further comprises acetylhexapeptide-8, and the natural complex extract may further comprise a Centella asiatica extract.

The “acetyl hexapeptide-8 (Acetyl Hexapeptide-8)” may be a reaction product of acetic acid and hexapeptide-8 (Ac-Glu-Glu-Met-Gln-Arg-Arg-NH ₂ ).

The " Centella asiatica L Urban " is a creeping perennial herb belonging to the Umbrella family, which is native to the African island of Madagascar, but is widely used in hot and humid places in the coastal areas of the Indian Ocean, northern Australia, and parts of Central and South America. In Korea, it grows wild on Jeju Island belonging to the temperate region and island regions in the southern region.

The cosmetic composition may further include niacinamide, adenosine, allantoin, tranexamic acid, and dipotassium glycyrrhizate. have.

In addition, the cosmetic composition may further include alpha-bisabolol, hydrolyzed glycosaminoglycan, sodium hyaluronate, hydrolyzed hyaluronic acid, and hyaluronic acid. , the complex components may exhibit synergistic activity with the components of the present invention.

The cosmetic composition may be for skin whitening or wrinkle improvement.

The "skin whitening" suppresses the production of various biomolecules such as melanin, oxidation-reduction hemoglobin, carotene, and melanoids that affect skin color, thereby alleviating skin pigmentation phenomena such as blemishes, freckles, and melasma. It refers to the effect of improving skin brightness and uniformity by alleviating redness.

The "wrinkle improvement" refers to a phenomenon that suppresses or inhibits the generation of wrinkles on the skin, or alleviates the already generated wrinkles.

The cosmetic composition may be for improving acne.

The “acne” is an inflammatory disease that occurs in the hair follicles and sebaceous glands of the skin, and refers to an inflammatory reaction caused by the accumulation of sebum in the hair follicles and the growth of acne bacteria therein.

The cosmetic composition can improve acne by inhibiting the acne-causing bacteria, thereby inhibiting clogging of pores and excessive secretion of sebum.

The cosmetic composition may be for skin moisturizing or skin texture improvement.

The "skin moisturizing" refers to any action that maintains the homeostasis of the skin tissue by appropriately controlling the loss of moisture (water evaporation) of the skin. The skin moisturizing effect may be accompanied by additional skin improvement effects in various aspects, such as a keratin improvement effect, a skin irritation reduction effect, and the like.

The “improvement of skin texture” may refer to smoothing and softening the skin surface by suppressing or inhibiting roughening of the skin surface due to internal or external influences such as aging and stress.

The cosmetic composition includes a softening lotion, a nourishing lotion, a moisture cream, a nourishing cream, a massage cream, a nourishing lotion, an essence, an ampoule, a gel, an eye cream, an oil, a foundation, a cleansing cream, a cleansing foam, a cleansing water, a shampoo, a conditioner, a pack, It may be formulated with one or more selected from the group consisting of sprays and powders.

The cosmetic composition may be formulated by a conventional method. In the formulation of external skin preparations, reference may be made to the contents disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, and in the formulation of cosmetic compositions, International cosmetic ingredient dictionary, 6th ed (The cosmetic, Toiletry and Fragrance Association) , Inc., Washington, 1995) may be referred to.

Specifically, the cosmetic composition can be prepared in general emulsified formulations and solubilized formulations. For example, lotion, such as a softening lotion or a nourishing lotion; emulsions such as facial lotions and body lotions; creams such as nourishing creams, moisturizing creams, and eye creams; essence; makeup ointment; spray; gel; pack; sunscreen; makeup base; foundations such as liquid type, solid type or spray type; powder; makeup removers such as cleansing creams, cleansing lotions, and cleansing oils; Alternatively, it may be formulated as a cleaning agent such as a cleansing foam, soap, body wash, but is not limited thereto.

In addition, the external preparation for skin may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.

In the cosmetic composition, in addition to the essential components in each formulation, other components may be appropriately blended within the range that does not impair the purpose according to the present invention, depending on the type or purpose of use, etc. of the formulation.

The cosmetic composition may include a conventionally acceptable carrier, for example, oil, water, surfactant, humectant, lower alcohol, thickener, chelating agent, colorant, preservative, fragrance, etc. may be appropriately formulated, but is limited thereto not.

The acceptable carrier may vary depending on the formulation. For example, when formulated into an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or their Mixtures can be used

When the cosmetic composition is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or mixtures thereof may be used as a carrier component, and in the case of a spray, chlorofluorohydro It may further contain a propellant such as carbon, propane, butane or dimethyl ether.

When the cosmetic composition is formulated as a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-butylglycol oil may be used.

When the cosmetic composition is formulated as a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier component, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like can be used.

The cosmetic composition is a fatty substance, organic solvent, solubilizer, thickening agent, gelling agent, softening agent, antioxidant, suspending agent, stabilizing agent, foaming agent, and fragrance commonly used in the industry according to the quality or function of the final product. , surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, commonly used in cosmetics It may additionally contain adjuvants commonly used in the field of cosmetics or dermatology, such as any other ingredients.

However, the adjuvant and its mixing ratio may be appropriately selected so as not to affect the desirable properties of the cosmetic composition according to the present invention.

Through the following examples, the present invention will be described in more detail, but it is obvious that the present invention is not limited by the following examples.

Examples and Comparative Examples

In order to verify the stability and functionality of the peptide complex of the present invention, samples were mixed as shown in Tables 1 and 2 below to set Examples and Comparative Examples. Each sample was obtained from the market and used.

Functional ingredient complex 1 was mixed with niacinamide, adenosine, allantoin, tranexamic acid, and dipotassium glycyrrhizate in the same ratio.

Functional ingredient complex 2 was mixed with alpha-bisabolol, hydrolyzed glycosaminoglycan, sodium hyaluronate, hydrolyzed hyaluronic acid, and hyaluronic acid in the same ratio.

[Content (parts by weight)] division Example One 2 3 4 5 6 7 8 SH-decapeptide-7 25 20 10 10 10 10 10 10 SH-Peptapeptide-19 25 20 10 10 10 10 10 10 Lotus callus culture extract 25 20 20 10 10 10 10 10 Pungran callus culture extract 25 20 20 10 10 10 10 10 Raspberry Ketone - 20 20 20 10 10 10 10 SH-octapeptide-4 - - 10 10 10 10 10 10 SH-oligopeptide-9 - - 10 10 10 10 10 10 Rice callus culture extract - - - 10 10 10 10 5 Resurrection plant callus culture extract - - - 10 10 10 5 5 Acetylhexapeptide-8 - - - - 10 5 5 5 Centella asiatica extract - - - - - 5 5 5 Functional Ingredient Complex 1 - - - - - - 5 5 Functional Ingredient Complex 2 - - - - - - - 5

[Content (parts by weight)] division comparative example One 2 3 4 5 6 7 8 9 10 SH-decapeptide-7 50 25 - - 20 - - - - - SH-Peptapeptide-19 50 25 - - 20 - - - - - Lotus callus culture extract - - 25 20 - 20 - - - - Pungran callus culture extract - - 25 20 - 20 - - - - Raspberry Ketone - - - 20 20 20 - - - - SH-octapeptide-4 - 25 - - 20 - - - - - SH-oligopeptide-9 - 25 - - 20 - - - - - Rice callus culture extract - - 25 20 - 10 - - - - Resurrection plant callus culture extract - - 25 20 - 10 - - - - Acetylhexapeptide-8 - - - - - 10 25 - - - Centella asiatica extract - - - - - 10 25 - - - Functional Ingredient Complex 1 - - - - - - 25 100 - 50 Functional Ingredient Complex 2 - - - - - - 25 - 100 50

Experimental Example 1: Skin safety test

In order to verify the skin safety of Examples and Comparative Examples, MTT assay experiments were performed on fibroblasts (HDF), melanocytes (B16F10), and keratinocytes (HaCaT).

After each of the samples of Examples and Comparative Examples were suspended in purified water at concentrations of 1, 5, 10, 50 and 100 mg/L, cell viability was measured.

Cytotoxicity was performed by modifying Mosmann's method for measuring cell viability using MTT {3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide} reagent.

Fibroblasts (HDF), melanocytes (B16F10), and keratinocytes (HaCaT) were each dispensed in a 96-well plate at a concentration of 1 × 10 ⁴ cells/well and cultured at 37° C., 5% CO ₂ for 48 hours. did.

After removing the culture medium and culturing the sample in a medium treated for each concentration for 48 hours, the medium was removed and washed repeatedly with PBS (Phosphate buffered saline).

MTT was dissolved in PBS at 5mg/mL, 50L was added, and incubated at 37°C, 5% CO ₂ for 48 hours. 100 L of DMSO (dimethyl sulfoxide) was added per well, and the absorbance was measured at 540 nm after stirring for 10 minutes.

As a result of the measurement, cytotoxicity was not confirmed at all concentrations of each sample. The above results suggest that the sample is not only harmless to the human body, but also has excellent human safety.

Experimental Example 2: Elastase inhibitory effect evaluation

The wrinkle improvement effect of the samples of Examples and Comparative Examples was evaluated through the inhibitory activity of elastase. The samples of Examples and Comparative Examples were suspended in purified water and elastase inhibitory activity was measured as follows.

1.6 mM elastase substrate (N-succinyl-(Ala)3-p-nitroanilide; SANA) and 0.2 mM Tris-HCl buffer (pH 8.0) were prepared. In a 96 well-plate, 0.165 mL of the Tris-HCl buffer, 0.005 mL of SANA and 0.01 mL of elastase were dispensed.

After putting the samples of Examples and Comparative Examples together with a positive control and reacting at 25° C. for 15 minutes, absorbance was measured at 405 nm.

The percentage (%) of elastase inhibitory activity was calculated as the ratio of the difference between the absorbance to which the sample was added compared to the absorbance to which the sample was not added.

Oleanolic acid was used as a positive control, and the concentration (IC ₅₀ ) that inhibited elastase by 50% of the samples of Examples and Comparative Examples was measured and shown in Table 3.

[Elastase 50% inhibitory concentration] division IC ₅₀ (μg/mL) Example 1 130.3 Example 2 126.7 Example 3 123.8 Example 4 121.2 Example 5 120.2 Example 6 116.1 Example 7 112.2 Example 8 102.2 Comparative Example 1 182.4 Comparative Example 2 204.8 Comparative Example 3 232.1 Comparative Example 4 199.3 Comparative Example 5 171.6 Comparative Example 6 198.4 Comparative Example 7 206.1 Comparative Example 8 194.0 Comparative Example 9 196.6 Comparative Example 10 204.6 positive control 95.8

The lower the IC ₅₀ value, the higher the elastase activity inhibition rate. Elastase is an enzyme that breaks down elastin, an important substrate for maintaining skin elasticity in the dermis.

In addition, elastase is a non-specific hydrolase capable of degrading collagen (collagen), which is another important matrix protein.

Therefore, the high elastase activity inhibition rate means that elastin and collagen, which are important for maintaining skin elasticity, are maintained for a long time, suggesting that the skin wrinkle improvement effect is excellent.

Referring to Table 3, the sample of Example inhibited elastase activity more effectively than Comparative Example, and the sample of Example was confirmed to have a synergistic action according to the combination of active ingredients.

Experimental Example 3: Wrinkle improvement effect evaluation

The skin wrinkle improvement effect was evaluated through the following experiment.

Adult women over 30 years of age were divided into 18 groups, and the test subject was thoroughly absorbed by applying the cream-formed cosmetic to the face after cleansing every morning and evening for 4 weeks.

ANTERA 3D (Miravex, Ireland) was applied to evaluate wrinkle improvement, and device measurements were made before and 3 weeks after use of the test substance.

The same test person measured the left corner of the eye of all subjects, and for reproducibility of the measurement, the same part was measured by overlapping the image measured before using the test substance.

The captured images were matched using ANTERA pro software, which is a dedicated software for ANTERA 3D, and then the matched measurement area was used for analysis.

The measured value was analyzed using the indentation index, which is a measurement variable, to analyze the Wrinkles small value indicating skin wrinkles.

Table 4 shows the amount of change in Wrinkles small value after 4 weeks of the test subject according to the use of cosmetics formulated with samples of Examples and Comparative Examples. The greater the wrinkle improvement effect, the greater the change in the Wrinkles small value.

division Wrinkles small average Example 1 1.44 Example 2 1.48 Example 3 1.52 Example 4 1.57 Example 5 1.58 Example 6 1.61 Example 7 1.66 Example 8 1.77 Comparative Example 1 0.84 Comparative Example 2 0.89 Comparative Example 3 0.76 Comparative Example 4 0.84 Comparative Example 5 0.97 Comparative Example 6 0.73 Comparative Example 7 0.76 Comparative Example 8 0.82 Comparative Example 9 0.77 Comparative Example 10 0.75

Referring to Table 4, the cosmetic formulated with the sample of Example was significantly superior in skin wrinkle improvement effect compared to Comparative Example, and the synergistic effect of the sample of Example was confirmed according to the combination of active ingredients.

Experimental Example 4: Evaluation of skin elasticity improvement effect

The skin elasticity improvement effect of the cosmetic compositions formulated with the samples of Examples and Comparative Examples was evaluated.

Each of the samples was formulated as a cream of the same composition, and only the samples of Examples and Comparative Examples were different.

The creams of Examples and Comparative Examples prepared by dividing into groups of healthy women (average age 37 years) over 20 years of age at a temperature of 24 to 26° C. and a humidity of 75% were applied, respectively.

The formulated cream was applied twice a day (morning and evening) around the eyes for 12 weeks, and skin elasticity was measured using a skin elasticity meter (Cutometer MPA580, Conrage + Khazaka, Germany).

The test results were described as the R8 (R8 (12 weeks)-R8 (0 weeks)) value of the cutometer MPA580, and the R8 value represents the property of skin viscoelasticity, and the results are shown in Table 5 below.

division Skin elasticity improvement effect Example 1 0.76 Example 2 0.78 Example 3 0.80 Example 4 0.83 Example 5 0.84 Example 6 0.85 Example 7 0.88 Example 8 0.94 Comparative Example 1 0.44 Comparative Example 2 0.47 Comparative Example 3 0.40 Comparative Example 4 0.44 Comparative Example 5 0.51 Comparative Example 6 0.39 Comparative Example 7 0.40 Comparative Example 8 0.43 Comparative Example 9 0.41 Comparative Example 10 0.40 Positive control (vitamin C) 0.86 negative control (no treatment) 0.31

Referring to Table 5, the sample of Example was significantly superior to the skin elasticity improvement effect compared to Comparative Example.

In particular, Example 1 including SH-decapeptide-7, SH-peptapeptide-19, lotus callus culture extract, and Pungran callus culture extract at the same time showed significantly superior skin elasticity improvement effect compared to the comparative example lacking some parts, The above results suggest a synergistic effect according to the combination of each composition.

Experimental Example 5: Evaluation of melanin production inhibitory activity

In order to evaluate the melanogenesis inhibitory effect of Examples and Comparative Examples, melanocytes B16F10 were cultured.

B16F10 was dispensed into each well of a 6-well plate to make 8 x 10 ⁴ cells, and then incubated for 18 hours in a cell incubator under 5% CO ₂ , 37°C conditions.

After 18 hours, the medium of the B16F10 cells was removed, and the samples of Examples and Comparative Examples were treated at the same concentration and cultured for 2 days in a cell incubator at 5% CO ₂ , 37° C. conditions.

After 2 days, cells were recovered from the plate using a scraper and centrifuged for 1 minute to obtain cells in the form of pellets.

1N NaOH was added to the obtained cells and reacted at 95° C. for 10 minutes to dissolve melanin.

After 10 minutes, melanin production was measured at a wavelength of 475 nm using a microplate reader, and the amount of protein was measured at a wavelength of 595 nm using a BCA assay.

Each experimental group was expressed numerically as “absolute amount of melanin / absolute amount of protein”, and the experimental group and control group were quantified relative to each other.

α-MSH was used as a melanogenesis inducer, and arbutin was used as a positive control.

division Melanin Relative Value (%) Example 1 55.7 Example 2 54.4 Example 3 53.1 Example 4 53.4 Example 5 52.7 Example 6 51.6 Example 7 50.1 Example 8 48.4 Comparative Example 1 72.7 Comparative Example 2 69.0 Comparative Example 3 76.5 Comparative Example 4 72.3 Comparative Example 5 61.6 Comparative Example 6 70.1 Comparative Example 7 69.6 Comparative Example 8 71.3 Comparative Example 9 70.7 Comparative Example 10 70.5 positive control 52.0 control 100.0

Referring to Table 6, the samples of Examples 1 to 8 were excellent in melanogenesis inhibitory activity compared to Comparative Examples 1 to 10.

In particular, Example 1 including SH-decapeptide-7, SH-peptapeptide-19, lotus callus culture extract, and Pungran callus culture extract at the same time had significantly superior melanin production inhibitory activity compared to the comparative example lacking some, The results suggest a synergistic effect according to the combination of each component of the present application.

Experimental Example 6: Evaluation of skin moisturizing activity

In Examples, the sample was suspended in purified water to evaluate the skin moisturizing effect.

Expression levels of HAS2 gene and AQP3 gene involved in skin moisturizing in human keratinocytes were measured at the mRNA level.

Human keratinocytes (HaCaT) were cultured with Dulbecco's Modified Eagle's Medium (DMEM), 10% Fatal bovine serum (FBS), and 1% Antibiotic-Antimycotic in a 100 mm/60.1 cm culture dish at 37°C and 5% CO ₂ conditions. did.

Keratinocytes were dispensed in a 96-well plate at a concentration of 1.0 x 10 ⁶ cells/mL, and cultured for 24 hours.

Samples of Examples and Comparative Examples diluted after exchange with DMEM-free medium were treated and cultured for 24 hours.

Cells were harvested, washed with chilled phosphate buffer (PBS), 1 mL of TRIzol™ reagent [TRIzol™ reagent, Life Technologies, Inc.] was added, and total RNA was extracted.

Changes in gene expression were measured by qRT-PCR (quantitative real time PCR), which binds a fluorescent material to a DNA product amplified in polymerase chain reaction (PCR) and continuously detects the fluorescent material.

To quantify changes in HAS2 and AQP3 gene expression, fluorescence values emitted by PCR products were measured through SYBR green I (Invitrogen).

A reaction solution was prepared by mixing 0.2 μM primer, 50 mM KCl, 20 mM Tris/HCl pH 8.4, 0.8 mM dNTP, 0.5U Extaq DNA polymerase, 3 mM MgCl ₂ , and 1×SYBR green in a PCR tube.

Primary denaturation was performed at 94°C for 3 minutes using Linegene K (BioER, China), followed by denaturation, annealing, and polymerization at 94°C for 30 seconds and 58°C for 30 seconds. Second, a total of 40 cycles were performed for 30 seconds at 72°C, and the fluorescence intensity was measured after each cycle was completed.

PCR results were verified with a melting curve for each result. After normalizing the threshold cycle (Ct) value of each gene to the Ct value of β, the amount of change in the Ct value was compared and analyzed.

The Ct value is the number of cycles when the amount of fluorescence (the number of amplified PCR products) generated by the PCR product reaches a constant reference value greater than or equal to the basic value, and the gene expression level can be confirmed through the Ct value.

The HAS2 and AQP3 mRNA expression level measurement results are shown in Table 7 below, and the mRNA expression level was measured after treatment with the sample at a concentration of 20 μg/mL in Example.

It can be evaluated that the higher the HAS2 and AQP3 expression promotion ability, the better the skin moisturizing effect.

[Fold of control] division HAS2 expression level AQP3 expression level Example 1 1.43 1.50 Example 2 1.48 1.55 Example 3 1.49 1.56 Example 4 1.54 1.62 Example 5 1.62 1.70 Example 6 1.64 1.72 Example 7 1.66 1.74 Example 8 1.68 1.76 Comparative Example 1 1.19 1.25 Comparative Example 2 1.13 1.18 Comparative Example 3 1.16 1.19 Comparative Example 4 1.11 1.21 Comparative Example 5 1.28 1.34 Comparative Example 6 1.21 1.24 Comparative Example 7 1.17 1.21 Comparative Example 8 1.14 1.23 Comparative Example 9 1.24 1.27 Comparative Example 10 1.27 1.30 negative control 1.00 1.00

Referring to Table 7, the expression levels of HAS2 gene and AQP3 gene increased according to the sample treatment of Examples.

The above results suggest that the sample of Examples can effectively enhance skin moisture by promoting the expression of aquaporin protein and the synthesis of hyaluronic acid according to the synergistic action of active ingredients.

Experimental Example 7: Evaluation of acne improvement effect

A group of 19 to 28-year-old women with acne was allowed to apply the formulated samples of Examples and Comparative Examples twice daily for 1 month to the face and the first half, and the degree of improvement was determined according to the user's opinion, and the results are shown in the table below 8 is shown.

As a result of the evaluation, it was confirmed that the sample of Example was significantly superior in the improvement of acne compared to the sample of Comparative Example.

division 2 weeks 4 weeks Example 1 + ++ Example 2 ++ ++ Example 3 ++ ++ Example 4 ++ +++ Example 5 ++ +++ Example 6 ++ +++ Example 7 ++ +++ Example 8 ++ +++ Comparative Example 1 ± + Comparative Example 2 ± ± Comparative Example 3 ± + Comparative Example 4 ± ++ Comparative Example 5 + ++ Comparative Example 6 ± + Comparative Example 7 ± + Comparative Example 8 + + Comparative Example 9 ± + Comparative Example 10 + +

<Evaluation criteria>

+++: There is a very good improvement effect / ++: There is a significant improvement effect / +: There is a slight improvement effect / ±: There is no improvement effect, but it does not deteriorate / -: There is no improvement effect, but rather weakened

Experimental Example 8: Evaluation of skin brightness improvement activity

After application to the skin using the samples of Examples and Comparative Examples, the skin brightness improvement effect was confirmed.

Cream formulations containing the samples of Examples and Comparative Examples were prepared, and the subjects were divided into 18 groups and applied to each, and then the change in skin brightness was confirmed.

After washing before taking a bath, a formulated cosmetic was applied, and it was used every day for 3 weeks. Skin brightness was checked with a LAB measuring device (Konica Minolta CR-300 Chroma Meter).

Then, the difference (L*) in skin color at the measurement time point and the application start time point after application of each test substance was calculated according to Equation 1 below, and the results are shown in Table 9 below.

The control group was formulated except for the samples of Examples and Comparative Examples, and arbutin was added to the positive control group.

The whitening effect is determined by comparing the L* between the sample application site and the control site. If the L* value is about 2, whitening of the deposited pigment is clear, and if it is about 1.5 or more, it can be determined that there is a whitening effect.

[Equation 1]

ΔL = L value at the time of measurement after application - L value at the beginning of application

division Brightness change after 3 weeks (ΔL) control - positive control 2.7 Example 1 2.3 Example 2 2.5 Example 3 2.6 Example 4 2.6 Example 5 2.6 Example 6 2.8 Example 7 2.9 Example 8 2.9 Comparative Example 1 1.7 Comparative Example 2 1.5 Comparative Example 3 1.4 Comparative Example 4 1.6 Comparative Example 5 2.0 Comparative Example 6 1.7 Comparative Example 7 1.8 Comparative Example 8 2.0 Comparative Example 9 1.9 Comparative Example 10 1.9

Referring to Table 9, when the cosmetic formulation formulated with the sample of Example was applied to the skin, skin brightness was remarkably improved after 3 weeks, and the sample of Example had an excellent skin brightness improvement effect compared to Comparative Example, so that the combination of active ingredients synergistic activity was confirmed.

Experimental Example 9: Evaluation of skin texture improvement activity

The skin texture improvement effect of the cosmetic compositions formulated with the samples of Examples and Comparative Examples was evaluated.

Each sample was formulated as a cream of the same composition, and the skin texture improvement effect was verified after dividing the subjects into 18 groups and applying each.

For a selected test subject among women aged 20 to 40, the skin of the test subject was measured before use using PRIMOS light (field of view 45, flexible 3D measuring, GFMesstechnik GmbH), and samples of Examples and Comparative Examples were formulated After using one cosmetic, the skin was measured again.

The measured value was expressed as an average value of three times except for the maximum and minimum values of Ra. The higher the measured value, the more uneven the skin is, and the evaluation results are shown in Table 10 below.

division Change after 4 weeks (Ra, Arbitrary unit) Example 1 13.44 Example 2 13.91 Example 3 14.01 Example 4 14.48 Example 5 15.23 Example 6 15.42 Example 7 15.60 Example 8 15.89 Comparative Example 1 11.19 Comparative Example 2 10.23 Comparative Example 3 10.65 Comparative Example 4 10.41 Comparative Example 5 11.59 Comparative Example 6 11.17 Comparative Example 7 10.81 Comparative Example 8 11.52 Comparative Example 9 11.26 Comparative Example 10 10.66

Referring to Table 10, the cosmetic composition formulated with the sample of Example significantly decreased the measured value after use, and thus the skin texture improvement effect was evaluated to be remarkably excellent, and synergistic activity according to the combination of active ingredients was confirmed.

The description of the present invention described above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a dispersed form, and likewise components described as distributed may also be implemented in a combined form.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present invention.

Claims (8)

a peptide complex comprising H-decapeptide-7 and H-peptapeptide-19;
A natural complex extract comprising a lotus callus culture extract and a fermented callus culture extract; and
A cosmetic composition comprising; raspberry ketone as an active ingredient. The method of claim 1,
The peptide complex further comprises SH-octapeptide-4 and SH-oligopeptide-9,
The natural complex extract further comprises a rice callus culture extract and Myrotamnus Flabellifolia callus culture extract, cosmetic composition. 3. The method of claim 2,
The peptide complex further comprises acetylhexapeptide-8,
The natural complex extract further comprises a Centella asiatica extract, cosmetic composition. 4. The method of claim 3,
Niacinamide, adenosine, allantoin, tranexamic acid, and further comprising dipotassium glycyrrhizate, cosmetic composition. 5. The method of claim 4,
Alpha-bisabolol, hydrolyzed glycosaminoglycan, sodium hyaluronate, hydrolyzed hyaluronic acid, and a cosmetic composition further comprising hyaluronic acid. 6. The method according to any one of claims 1 to 5,
A cosmetic composition for skin whitening or wrinkle improvement. 6. The method according to any one of claims 1 to 5,
A cosmetic composition for improving acne. 6. The method according to any one of claims 1 to 5,
A cosmetic composition for moisturizing or improving skin texture.