KR102022216B1 - Antiaging cosmetic composition comprising phytoplacenta complex extract - Google Patents

Abstract

The present invention relates to an anti-aging cosmetic composition comprising a phytoplacenta complex extract of beans, lotus, peas, magnolia, camellia, plum and peony. The composition according to the present invention shows a strong antioxidant effect and an effect of protecting the skin from aging factors such as, various oxidative toxicity and phototoxicity and thus can be favorably used as a functional anti-aging cosmetic composition.

Description

Anti-aging cosmetic composition containing phytoplacenta complex extract {ANTIAGING COSMETIC COMPOSITION COMPRISING PHYTOPLACENTA COMPLEX EXTRACT}

The present invention relates to a cosmetic composition comprising a phytoPlacenta complex extract, and more particularly to an anti-aging cosmetic composition comprising a phytoPlacenta complex extract of beans, lotus, peas, magnolia, camellia, plum and peony.

The aging process of the skin involves free radicals, which correspond to free radicals. Free radicals play an important role in eliminating or inhibiting invading substances in the body, such as pathogens or viruses, while free radicals are enzymatic and non-enzymatic antioxidant defense systems when overproduced because of their high energy and reactivity. Is known to accelerate skin aging (Yang et al., J. Soc. Cosmet. Sci. Korea, 34: 275-286, 2008). Free radicals and free radicals are produced more when the skin is exposed to ultraviolet light, and thus it is reported to cause phototoxicity, photosensitivity, skin aging and various immune-related diseases (Norins, J. Invest. Dermatol., 39: 445, 1962; Cadenas, Ann. Rev. Biochem., 58:79, 1989).

In the skin aging process, lipids, proteins, polysaccharides, nucleic acids, and the like, which are major constituents of the skin, are oxidized, thereby destroying skin cells and tissues. In particular, oxidation of proteins damages skin connective tissues such as collagen, elastin, proteoglycans, fibronectin, hyaluronic acid, resulting in excessive inflammatory reactions and reduced skin elasticity.

Therefore, in order to quickly recover the aged skin and prevent the aging of the skin, it is important to protect the cell membrane by eliminating active oxygen mediated by the body's metabolic process and ultraviolet irradiation.

And the like (superoxide anion radical), OH ( hydroxyl radical), 1 O 2 (singlet oxygen), H 2 O 2 (hydrogen peroxide) - free radical and active oxygen species (reactive oxygen species, ROS) is typically O ₂ . Antioxidants for preventing skin aging by inhibiting the enzymatic and non-enzymatic oxidation by the reactive oxygen species have been developed in various ways, can be divided into synthetic antioxidants and natural antioxidants.

Representative examples of the synthetic antioxidants include butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA) and the like, which have excellent antioxidant effects but have a high carcinogenic problem, which may adversely affect the human body. Compared to the known enzymatic antioxidant effect is not good.

Recently, researches on natural antioxidants that can supplement the problems of synthetic antioxidants have been actively conducted. For example, Korean Patent Publication No. 10-1773456 (August 31, 2017) contains a combination of ginseng, Hyunsam, Dansam, Gosam, Sasam, Mansam, and Dannersam, and fermented herbal extracts fermented sequentially using five microorganisms. It is disclosed that there is an antioxidant, skin wrinkle improvement and skin inflammation relieving effect. Although natural antioxidants have less side effects such as carcinogenicity than synthetic antioxidants, they still show limitations in meeting both enzymatic and non-enzymatic antioxidant effects.

In addition, in addition to free radicals, aging involves an enzyme called matrix metalloproteinase (MMP). In healthy skin, the synthesis and degradation of extracellular matrix such as collagen is properly regulated in vivo, whereas as aging progresses, the synthesis of collagen decreases and the elasticity of the skin decreases and wrinkles are formed. At this time, since the MMP enzyme has a function of degrading collagen, when the expression of MMP is promoted by ultraviolet irradiation or the like, aging of the skin is accelerated.

On the other hand, animal-derived placental ingredients have been used as raw materials for cosmetics due to the effects of strong skin regeneration, elasticity regeneration, and moisturizing, but may cause disease infection, skin irritation and / or allergy phenomenon, and as an alternative to this recently, There is a lot of research and development on the placenta of plants (phytoplacenta). Korean Patent Publication No. 10-1549299 (2015.09.01) discloses that the composition containing the lotus cell culture of lotus plant or its extract as an active ingredient has excellent skin collagen synthesis ability without being toxic to skin cells, and shrinking pores and whitening. It has been shown to have sebum secretion, moisturizing, anti-inflammatory and acne-improving effects. In addition, Korean Patent Registration No. 10-1810410 (December 19, 2017) shows that the mixed culture extract extracted by mixing and cultivating Cheongho wheat pollen with mushroom mycelium exhibits excellent anti-aging, skin moisturizing, skin wrinkle and elasticity improvement and skin cell regeneration activity. It is disclosed that it can.

As described above, in order to develop a highly functional natural anti-aging cosmetics, a new natural substance, a new combination of natural ingredients, a blending ratio, and a method of producing a powerful antioxidant effect and having no side effects and at the same time efficiently suppressing MMP expression There is a continuing need for research.

Registered Patent Publication No. 10-1773456 (2017.08.31) Registered Patent Publication No. 10-1549299 (2015.09.01) Registered Patent Publication No. 10-1810410 (2017.12.19)

The present inventors are aware of the problems of the prior art and can exhibit the scavenging efficacy of free radicals that are a direct cause of aging among the various mechanisms associated with skin aging and trigger the operation of other mechanisms to develop high functional anti-aging cosmetic compositions. As a result of long experiments with phytoPlacenta, the present invention has been achieved.

According to one embodiment of the present invention, there is provided an anti-aging cosmetic composition comprising a phytoPlacenta complex extract of beans, lotus, pea, magnolia, camellia, plum and peony.

As used herein, the term "soybean" is an annual herbaceous plant belonging to the legume family, the scientific name is Glycine max. It can be frosted, seomoktae, blacktae, yellow, green pea (cheongtae), kidney beans, green beans, hedge beans, green beans, red beans, soybeans, bean sprouts beans, sesame beans or soybeans, the type is not limited. Beans are grown in Korea, China, and Japan.

As used herein, the term “lotus flower” is a perennial aquatic plant belonging to the family Lotus, and its scientific name is Nelumbo nucifera. Lotus is grown in Korea and India.

As used herein, the term "peas" is a biennial herbaceous plant belonging to the legume, scientific name Pisum sativum. Peas are grown in China, Russia, and Korea.

The term "magnolia" as used herein is a deciduous textbook belonging to the Magnolia family, and its scientific name is Magnolia kobus. Magnolia is grown in Korea, China and Japan.

As used herein, the term "camellia" is an evergreen tree of the family Tea Tree, the scientific name is Camellia Japonica. Camellia is grown in Korea, China, and Japan.

As used herein, the term “plum” is a deciduous arborescent of the Rosaceae family, and its scientific name is Prunus Mume. Plums are grown in Korea and China.

As used herein, the term "peony" is a deciduous shrub belonging to the peony family, scientific name is Paeonia suffruticosa. Also called a neck. Peonies are grown in Korea, China, Japan and France.

As used herein, the term "phytoplacenta" refers to a site that manages seed growth of a plant, and refers to a site where seeds are attached in an embryonic region or an ovary of pistil.

As used herein, the term "extract" refers to a result obtained by extraction with an appropriate extraction solvent, which includes an extract obtained by extraction treatment, a dilution or concentrate of the extract, a dried product obtained by drying the extract, a crude product thereof, or It is understood that a purified product, or fraction, is included. Preferably, phytoplacental complexes of beans, lotus, peas, magnolia, camellia, plums and peonies (for example, in the form of dry forms of phytoplacens of beans, lotus, peas, magnolia, camellia, plums and peonies) By extracting, it is possible to obtain a phytoplacenta complex extract according to the present invention.

The beans, lotus, pea, magnolia, camellia, plum and peony phytoplacenta are 1 to 91% by weight, 1 to 91% by weight of lotus, 1 to 91% by weight of pea, 1 to 91% by weight, respectively, based on the total weight of the complex. 91 weight percent, camellia 1-91 weight percent, plum 1-91 weight percent, peony 1-91 weight percent.

Extract according to the present invention can be prepared using a common extraction method known in the art, for example, hot water extraction, hot water extraction, cold needle extraction, reflux cooling extraction, ultrasonic extraction, hot pressurized extraction, etc. can be used. have.

Extraction solvents used to prepare the extract according to the present invention include water; Lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol, and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; Hydrocarbon solvents such as chloroform, methyl acetate, ethyl acetate, benzene, hexane, diethyl ether, petroleum ether and dichloromethane; Or mixed forms thereof, but is not limited thereto.

The cosmetic composition according to the present invention includes phytoplacenta complex extract of soybean, lotus, pea, magnolia, camellia, plum and peony as active ingredients. The content of the extract in the cosmetic composition may be suitably determined in consideration of various factors such as the purpose of use, duration of use, type of formulation, route of administration, and skin condition of the subject. Preferably, the content of the extract is 0.0005 to 10% by weight relative to the total weight of the cosmetic composition. Bean, lotus, pea, magnolia, camellia, plum and phyto placenta complex extract of the peony as the active ingredient may be a complex of the weight ratio of 1 to 3: 1 to 2: 1: 1: 1: 1: 1. For example, a weight ratio of 1: 1: 1: 1: 1: 1: 1, or a weight ratio of 2: 1: 1: 1: 1: 1: 1, or 3: 2: 1: 1: 1: 1: It may be that the composite of the weight ratio of 1.

The cosmetic composition according to the present invention exhibits an excellent effect of protecting the skin from aging factors such as antioxidant effect, various oxidative toxicity and phototoxicity. In particular, the cosmetic composition according to the present invention exhibits a very good anti-aging effect by affecting various mechanisms of skin aging.

As used herein, the term "prevention" may be interpreted to include the acts of delaying or alleviating, as well as the acts of inhibiting or inhibiting the parameters associated with the skin condition, such as the extent of symptoms or progression.

The cosmetic composition according to the present invention can be prepared in the form of a general emulsion formulation, solubilized formulation, etc. using a conventionally known manufacturing method. At this time, the cosmetic composition of the present invention, patches, ointments, skin adhesive gels, creams, packs, lotions, essences, sprays, masks, foundations, makeup bases, detergents, water (W) type, Oil (O), silicon (S), oil in water (O / W), water in water (W / O), silicon in water (W / S), silicon in water (S / W), solid, It can be prepared in a variety of formulations, such as liquid, it may be applied to a cosmetic manufacturing method commonly used.

The cosmetic composition according to the present invention may further comprise additional ingredients in addition to the extract for enhancing efficacy. For example, the additional ingredient is not limited unless it counteracts or reduces the efficacy of the extract. Optionally, it may further include adjuvants, carriers, and the like, commonly used in the cosmetic field. Optionally, ingredients typically used to add or enhance cosmetic function may also be added. For example, stabilizers, emulsifiers, thickeners, moisturizers, liquid crystal film strengthening agents, pH regulators, antibacterial agents, water-soluble polymers, coating agents, metal ion sequestrants, amino acids, organic amines, polymer emulsions, pH regulators, skin nutrients, antioxidants, One or more aqueous additives selected from antioxidants, preservatives, fragrances and the like; And one or more oily additives selected from fats and oils, waxes, hydrocarbon oils, higher fatty acid oils, higher alcohols, synthetic ester oils, silicone oils, and the like.

The active ingredient of the cosmetic composition according to the present invention can be prepared through the step of extracting the active substance by adding a solvent to the phytoplacenta complex of beans, lotus, peas, magnolia, camellia, plum and peony.

Optionally, separating and filtering the extract; Primary concentration step of concentrating the extract; It may further comprise one or more of the secondary concentration steps to obtain a high content of the active substance. The first concentration step and the second concentration step may be used for the solvent removal action. The first concentration step may be, for example, heat drying, reduced pressure drying, spray drying, freeze drying, but is not limited thereto.

The extraction process may be carried out by adding a phytoPlacenta complex to the solvent at a rate of 0.5 to 20.0% to extract for 0.5 to 5 hours.

The extract included in the cosmetic composition according to the present invention may be in a dried form, a concentrated form, a solution form and the like.

Soybean, lotus, pea, magnolia, camellia, plum and peony phytoplacenta complex extract according to the present invention shows a significantly superior effect than the individual extract. In particular, the cosmetic composition comprising a complex extract consisting of a specific composition ratio according to the present invention shows a significant synergistic effect compared to other composition ratios.

Specifically, the composition according to the present invention has a very high content of representative antioxidant components such as phenolic compounds and flavonoids. In particular, the free radical scavenging test not only showed excellent non-enzymatic antioxidant effects, but also the active oxygen scavenging test demonstrated excellent enzymatic antioxidant effects. In addition, the extract according to the present invention does not cause side effects such as skin irritation and carcinogenicity, which are represented by synthetic antioxidants, as the components are all made of natural substances.

Moreover, when irradiated with ultraviolet light, one of the main causes of skin aging, it can alleviate the cytotoxicity caused by ultraviolet light.

In addition, collagen degrading enzyme MMP (Matrix metalloproteinase) is promoted by ultraviolet irradiation, but the extract according to the present invention can significantly inhibit the expression of MMP, thereby effectively preventing the breakdown of collagen to improve skin elasticity Improve and inhibit aging progression.

1 is a graph showing the cytotoxicity relaxation rate (%) by the ultraviolet rays represented by each sample.
2 is a graph showing MMP-1 expression inhibition rate (%) indicated by each sample.

Hereinafter, specific examples will be described for better understanding of the present invention. However, the following examples should not be construed as limiting the present invention, and conventional variations of those skilled in the art are possible within the scope of the present invention.

Example 1

Soybeans, lotus, peas, magnolia, camellia, plum and peony embryo (seed) portions were stripped with razor blades to obtain respective phytoplacentas. The obtained phytoplacenta complex (1 part by weight of beans, 1 part by weight of lotus, 1 part by weight of peas, 1 part by weight of magnolia, 1 part by weight of camellia, 1 part by weight of plum, 1 part by weight of peony) in a 70% aqueous ethanol solution in 10% by weight Was added in the amount of. Subsequently, the immersed phytoplacenta complex was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 1" and used in the following test example.

Example 2

Soybeans, lotus, peas, magnolia, camellia, plum and peony embryo (seed) portions were stripped with razor blades to obtain respective phytoplacentas. The obtained phytoplacenta complex (2 parts by weight of beans, 1 part by weight of lotus, 1 part by weight of peas, 1 part by weight of magnolia, 1 part by weight of camellia, 1 part by weight of plum, 1 part by weight of peony) in a 70% aqueous ethanol solution in 10% by weight Was added in the amount of. Subsequently, the immersed phytoplacenta complex was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 2" and was used in the following test example.

Example 3

Soybeans, lotus, peas, magnolia, camellia, plum and peony embryo (seed) portions were stripped with razor blades to obtain respective phytoplacentas. The obtained phytoplacenta complex (3 parts by weight of beans, 2 parts by weight of lotus, 1 part by weight of pea, 1 part by weight of magnolia, 1 part by weight of camellia, 1 part by weight of plum, 1 part by weight of peony) in 70% ethanol aqueous solution 10% by weight Was added in the amount of. Subsequently, the immersed phytoplacenta complex was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 3" and was used in the following test example.

Comparative Example 1

Dried bean seeds were added to a 70% aqueous ethanol in an amount of 10% by weight. Subsequently, the soaked beans were extracted for 2 hours under a temperature condition of 80 ℃ in a reflux extractor equipped with a cooling capacitor. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 4" and was used in the following test example.

Comparative Example 2

The embryo (sea) part of the bean was stripped with a razor blade to obtain a phytoplacenta. The soybean phytoplacenta obtained was added to an aqueous 70% ethanol solution in an amount of 10% by weight. Subsequently, the soaked phytoplacenta was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 5" and was used in the following test example.

Comparative Example 3

The dried lotus flower was added to the 70% aqueous ethanol in an amount of 10% by weight. Subsequently, the immersed lotus was extracted for 2 hours under a temperature condition of 80 ℃ in a reflux extractor equipped with a cooling capacitor. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 6" and was used in the following test example.

Comparative Example 4

The embryo (sea) part of the lotus was stripped with a razor blade to obtain a phytoplacenta. The obtained lotus phytoplacenta was added to an aqueous 70% ethanol solution in an amount of 10% by weight. Subsequently, the immersed lotus phytoplacenta was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 7" and was used in the following test example.

Comparative Example 5

Dried peas were added to an aqueous 70% ethanol solution in an amount of 10% by weight. Subsequently, the soaked peas were extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 8" and was used in the following test example.

Comparative Example 6

The embryo (seed) portion of the pea was stripped with a razor blade to obtain a phytoplacenta. The obtained pea phytoplacenta was added to an aqueous 70% ethanol solution in an amount of 10% by weight. Subsequently, the soaked pea phytoplacenta was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 9" and was used in the following test example.

Comparative Example 7

Dried magnolia was added to the 70% aqueous ethanol in an amount of 10% by weight. Subsequently, the immersed magnolia was extracted for 2 hours under a temperature condition of 80 ℃ in a reflux extractor equipped with a cooling capacitor. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 10" and was used in the following test example.

Comparative Example 8

The embryonic (seed) portion of the magnolia was stripped with a razor blade to obtain a phytoplacenta. The obtained magnolia phytoplacenta was added to 70% aqueous ethanol in an amount of 10% by weight. Subsequently, the immersed magnolia phytoplacenta was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 11" and was used in the following test example.

Comparative Example 9

Dried camellia was added to an aqueous 70% ethanol solution in an amount of 10% by weight. Subsequently, the immersed camellia was extracted for 2 hours under a temperature condition of 80 ° C. in a reflux extractor equipped with a cooling capacitor. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 12" and was used in the following test example.

Comparative Example 10

The embryonic (eye) part of the camellia was stripped with a razor blade to obtain a phytoplacenta. The camellia phytoplacenta was added to an aqueous 70% ethanol solution in an amount of 10% by weight. Subsequently, the immersed camellia phytoplacenta was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 13" and was used in the following test example.

Comparative Example 11

Dried plums were added to an aqueous 70% ethanol solution in an amount of 10% by weight. Subsequently, the immersed plums were extracted for 2 hours under a temperature condition of 80 ℃ in a reflux extractor equipped with a cooling capacitor. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 14" and was used in the following test example.

Comparative Example 12

An embryo (sea) part of the plum was stripped with a razor blade to obtain a phytoplacenta. The resulting plum phytoplacenta was added to the 70% aqueous ethanol in an amount of 10% by weight. Subsequently, the immersed plum phytoplacenta was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 15" and was used in the following test example.

Comparative Example 13

Dried peony was added to the 70% aqueous ethanol in an amount of 10% by weight. Subsequently, the immersed peony was extracted for 2 hours under a temperature condition of 80 ℃ in a reflux extractor equipped with a cooling capacitor. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 16" and was used in the following test example.

Comparative Example 14

The embryo (sea) part of the peony was stripped with a razor blade to obtain a phytoplacenta. The resulting peony phytoplacenta was added to the 70% aqueous ethanol in an amount of 10% by weight. Subsequently, the immersed peony phytoplacenta was extracted in a reflux extractor equipped with a cooling capacitor for 2 hours under a temperature condition of 80 ° C. After the extraction was completed, it was filtered through a 80 mesh, 1.2 μm filter. The filtrate thus obtained was concentrated using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) for 1 hour at 65 ℃, 20 hPa conditions.

The resulting concentrated powder was referred to as "Sample 17" and was used in the following test example.

Test Example 1 Content of Antioxidant Components

With respect to the extracts prepared in Examples and Comparative Examples, the content of the antioxidant component was confirmed. To this end, the content of phenolic compounds and flavonoids, which are well known in the art to exhibit antioxidant effects, were measured in each sample.

First, the total content of phenolic compounds was measured as follows.

10 mL of distilled water was added to 1 mL of each sample, 2 mL of Folin-Ciocalteu phenol reagent (Sigma) was added, mixed, and reacted at room temperature for 5 minutes. 2 mL of 20% sodium carbonate was added thereto, mixed and allowed to react at room temperature for 1 hour. Thereafter, absorbance was measured at 680 nm using a microplate reader (UVT-06685, Thermo max, USA). In this case, gallic acid (gallic acid, Sigma, USA) was used as the indicator, and the total content of the phenolic compound was calculated by substituting absorbance values of each sample by creating a calibration curve for each concentration of the indicator. The results are shown in Table 1 below.

Total Phenolic Content (mg / g Extract) Sample 1 205.3 ± 7.6 Sample 2 234.2 ± 2.3 Sample 3 259.8 ± 0.7 Sample 4 128.5 ± 2.1 Sample 5 173.1 ± 3.2 Sample 6 131.9 ± 0.5 Sample 7 163.4 ± 7.1 Sample 8 99.9 ± 0.8 Sample 9 121.1 ± 1.2 Sample 10 106.4 ± 2.2 Sample 11 145.9 ± 3.2 Sample 12 111.1 ± 6.3 Sample 13 145.1 ± 3.4 Sample 14 60.1 ± 0.6 Sample 15 88.9 ± 0.7 Sample 16 103.7 ± 5.5 Sample 17 142.3 ± 1.8

Next, the total content of flavonoids was measured as follows.

To 1.5 mL of each sample, 2% AlCl ₃ .6H ₂ O dissolved in the same amount of methanol was mixed and reacted at room temperature for 10 minutes. Thereafter, absorbance was measured at 367 nm using a microplate reader (UVT-06685, Thermo max, USA). At this time, catechin (catechin, Sigma, USA) was used as a marker, and the total content of flavonoids was calculated by substituting absorbance values of each sample by creating a calibration curve for each concentration of the marker. The results are shown in Table 2 below.

Total Flavonoid Content (mg / g Extract) Sample 1 200.0 ± 2.8 Sample 2 218.1 ± 7.2 Sample 3 234.7 ± 1.9 Sample 4 110.5 ± 0.1 Sample 5 123.1 ± 2.1 Sample 6 103.4 ± 3.5 Sample 7 132.6 ± 1.8 Sample 8 99.9 ± 4.8 Sample 9 121.1 ± 3.0 Sample 10 94.2 ± 1.2 Sample 11 103.4 ± 2.7 Sample 12 91.1 ± 4.4 Sample 13 102.3 ± 0.4 Sample 14 84.1 ± 0.9 Sample 15 94.8 ± 4.3 Sample 16 70.6 ± 3.5 Sample 17 80.7 ± 5.2

Through the test results of Table 1 and Table 2, it was confirmed that even in the same natural extract, the content of the phenolic compound and the flavonoids in the sample extracted phytoplacenta site compared to other sites. Surprisingly, the phytoPlacenta complex extracts (Samples 1 to 3) showed significantly higher phenolic compounds and flavonoid contents compared to the phytoPlacenta extracts (Samples 5, 7, 9, 11, 13, 15 and 17) of the single substance. Could. This means that the antioxidant power of the extract according to the present invention is remarkably large. In particular, the phytoflavent complex extract of Sample 3 had the highest phenolic compound and flavonoid content.

Test Example 2 Antioxidant Effect

In order to assay the skin aging improvement effect of the present invention, to determine the antioxidant capacity of each sample. To this end, free radical scavenging test (non-enzymatic antioxidant effect test) and active oxygen scavenging test (enzymatic antioxidant effect test) were conducted.

(1) Free radical scavenging test (non-enzymatic antioxidant effect test)

The free radical scavenging test frequently used in the art to confirm the antioxidant effect is based on the principle that the absorbance of stable 2,2-diphenyl-1-picryl-hydrazyl (DPPH) shows a maximum at 540 nm. Thus, as free radical DPPH is erased by the sample and becomes purple to transparent color, i.e. as the free radical scavenging rate is increased, the absorbance at 540 nm decreases.

First, 1 mL of a solution in which the samples prepared in Examples and Comparative Examples were diluted in methanol was mixed with 1 mL of 0.1 mM DPPH (Sigma) solution. Subsequently, after standing at 37 ° C. for 15 minutes, the absorbance was measured at 540 nm using a microplate reader (UVT-06685, Thermo max, USA).

In the control group, 1 ml of DPPH and 1 ml of methanol were measured to measure absorbance in the same manner, and 1 ml of methanol and 1 ml of sample were added to obtain color correction values of the sample and the control. The free radical removal rate was calculated by the following Equation 1, and in Table 3, SC ₅₀ indicates the concentration of the sample required to remove 50% of the free radicals.

[Equation 1]

% Free radical scavenging ratio = 100-((absorbance of sample / absorbance of control) x 100)

Free radical scavenging test (SC ₅₀ , μg / ml) Sample 1 32.6 ± 4.3 Sample 2 28.1 ± 1.8 Sample 3 16.7 ± 2.3 Sample 4 84.3 ± 3.5 Sample 5 67.5 ± 1.1 Sample 6 85.6 ± 2.2 Sample 7 71.4 ± 4.3 Sample 8 100.3 ± 1.7 Sample 9 84.6 ± 0.8 Sample 10 113.8 ± 7.9 Sample 11 91.0 ± 3.7 Sample 12 123.7 ± 0.8 Sample 13 100.3 ± 3.8 Sample 14 102.1 ± 3.6 Sample 15 91.9 ± 4.0 Sample 16 101.9 ± 2.1 Sample 17 89.9 ± 0.2

In Table 3, the smaller the SC ₅₀ value means that the antioxidant activity is higher. The SC ₅₀ values of Samples 1 to 3 of the Examples were significantly smaller than the SC ₅₀ values of Samples 4 to 17 of the Comparative Examples, which means that the antioxidant activity of the phytoPlacenta complex extract according to the present invention is excellent. . In particular, SC ₅₀ of Sample 3 showed the smallest value.

(2) free radical scavenging test (enzymatic antioxidant effect test)

The free radical scavenging test is based on the principle that free radicals are generated by the enzymatic reaction of xanthine / xanthine oxidase (Sigma). The scavenging ability of active oxygen can be confirmed by measuring the change in absorbance using oxidation of nitroblue tetrazolium (NBT) by active oxygen.

2.4 mL of Na ₂ CO ₃ , 0.1 mL of Xantine (Sigma), 0.1 mL of EDTA, 0.1 mL of BSA (Bovine Serum Albumin, Sigma), 0.1 mL of NBT and 0.1 mL of sample were mixed, and a vortex mixer (Type 37600 Mixer, Mini mix, USA) and then left at 25 ° C. for 10 minutes for voltexing. Subsequently, 0.1 mL of xanthine oxidase was added thereto and reacted at 25 ° C. for 20 minutes. 6 mM CuCl ₂ was added to stop the reaction, and the absorbance was measured at 540 nm using a microplate reader (UVT-06685, Thermo max, USA).

In the control group, the absorbance was measured in the same manner by adding tertiary distilled water, and the third distilled water was added instead of the xanthine oxidase solution.

The oxygen free removal rate was calculated by Equation 2 below, and in Table 4, IC ₅₀ indicates the concentration of the sample required to remove 50% of the free radicals.

[Equation 2]

% Of free radicals = 100-((absorbance of sample / absorbance of control) x 100)

Free radical scavenging test (IC ₅₀ , μg / ml) Sample 1 58.4 ± 0.8 Sample 2 43.2 ± 7.8 Sample 3 25.4 ± 3.0 Sample 4 92.9 ± 1.3 Sample 5 79.4 ± 6.6 Sample 6 97.0 ± 2.2 Sample 7 82.3 ± 3.9 Sample 8 113.8 ± 0.7 Sample 9 99.6 ± 3.4 Sample 10 121.7 ± 3.1 Sample 11 90.0 ± 0.1 Sample 12 115.9 ± 3.6 Sample 13 97.8 ± 5.6 Sample 14 120.0 ± 9.4 Sample 15 104.4 ± 4.4 Sample 16 115.8 ± 0.7 Sample 17 89.0 ± 8.2

In Table 4, the smaller the IC ₅₀ value means that the antioxidant activity is higher. The IC ₅₀ values of Samples 1 to 3 of the Examples showed significantly smaller values than the IC ₅₀ values of Samples 4 to 17 of the Comparative Examples, which means that the antioxidant activity of the phytoplacenta complex extract according to the present invention is excellent. . In particular, the IC ₅₀ of Sample 3 showed the smallest value.

Test Example 3 Cytotoxic Alleviation Effect by UV Irradiation

In order to confirm the cytotoxic alleviation effect by UV irradiation of the phytoplacenta complex extract according to the present invention was tested as follows.

Fibroblasts were dispensed 1 × 10 ⁵ into 24-well plates and attached for 24 hours. Each well was washed once with PBS and 500 μL of PBS was added to each well. Then, 10 mJ / cm <^2> of ultraviolet-rays were irradiated to fibroblasts using the ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki, Japan). Then PBS was removed and 1 mL of cell culture medium (DMEM with 10% FBS) was added.

Each sample produced above was treated with a concentration of 0.01%, and then incubated for 24 hours. The medium was then removed and 500 μL of cell culture medium and 60 μL of MTT solution (2.5 mg / mL) were added per well, followed by incubation in a 37 ° C. CO ₂ incubator for 2 hours. The medium was removed and 500 μL of isopropanol-HCl (0.04 N) was added. The cells were lysed by shaking for 5 minutes and 100 μL of the supernatant was transferred to a 96-well test plate. Thereafter, the absorbance at 565 nm was measured using a microplate reader (UVT-06685, Termo max, USA).

Cell survival rate (%) was calculated by Equation 3 below, and cytotoxicity relaxation rate (%) by UV was calculated by Equation 4 below. Based on the sample treatment concentration 0.01%, the cytotoxicity relaxation rate (%) by ultraviolet rays is shown in Table 5 below and also graphically shown in FIG.

[Equation 3]

Cell survival rate (%) = ((St-Bo) / (Bt-Bo)] x 100

Bo: 565 nm absorbance of the wells, only by color reaction of the cell culture medium

Bt: 565 nm absorbance of the wells, which developed and reacted the wells untreated sample

St: 565 nm absorbance of the wells, the color reaction of the wells treated with the sample

[Equation 4]

Reduction rate of cytotoxicity by ultraviolet ray (%) = [1- (St-Bo) / (Bt-Bo)] x 100

Bo: cell viability of wells without UV irradiation

Bt: cell viability of wells that have not been irradiated with UV

St: Cell survival rate of wells that were irradiated with UV

Cytotoxic alleviation rate (%)
Sample 1 22.4 Sample 2 30.9 Sample 3 35.8

As shown in Table 5, it was confirmed that the extract according to the present invention has an effect of alleviating the cytotoxicity caused by ultraviolet light. In particular, sample 3 showed a markedly high cytotoxic alleviation rate.

Test Example 4 Effect of Inhibiting MMP-1 Expression After Ultraviolet Irradiation

As mentioned above, the matrix metalloproteinase (MMP), which degrades collagen, can be promoted by UV irradiation, in which case collagen degradation is activated, resulting in decreased elasticity of the skin and wrinkle formation. This leads to.

In this test example, the effect of inhibiting MMP-1 expression by the sample was confirmed by measuring the concentration after UV irradiation and sample addition of MMP-1 related to skin elasticity reduction, wrinkle formation, and aging. To this end, enzyme immunoassay (ELISA) was performed as follows.

Human dermal fibroblasts were irradiated with UVA at an energy of 5 J / cm ² using a UV chamber. In order to determine the amount of UV irradiation and incubation time, preliminary experiments confirmed the conditions for the maximum amount of MMP expression in fibroblasts. Negative controls were wrapped in tinfoil and maintained for the same time in a UVA environment. UVA emission was measured using a UV radiometer. The cells during the UVA irradiation were previously dispensed, and then irradiated with UVA, exchanged with the medium containing the sample, and then cultured for 24 hours, and the medium was recovered and coated in 96-well.

MMP-1 (Ab-5) monoclonal antibody and MMP-2 (Ab-3) monoclonal antibody were treated as primary antibodies and reacted at 37 ° C. for 60 minutes. After reacting the secondary antibody anti-mouse IgG (whole mouse, alkaline phosphatase conjugated) for 60 minutes, the alkaline phosphatase substrate solution (1 mg / ml ρ-nitrophenyl phosphate in diethanolamine buffer) for 30 minutes at room temperature. Then, absorbance at 405 nm was measured using a microplate reader (UVT-06685, Termo max, USA).

At this time, a sample without addition of a sample was used as a negative control, and retinol, which was known to be excellent in inhibiting MMP-1 expression, was used as a positive control. Table 6 shows the inhibition rate (%) of MMP-1 expression based on 0.01% sample concentration.

MMP-1 expression inhibition rate (%)
Sample 1 32.8 Sample 2 38.4 Sample 3 42.3 Positive Control-Retinol 22.5

As shown in Table 6, Samples 1 to 3 according to the present invention showed a significantly higher value compared to the inhibition rate of MMP-1 expression of the retinol used as a positive control, especially in the case of Sample 3 22.5% The effect was nearly twice that of. This means that the extract according to the present invention can effectively inhibit collagen degradation by inhibiting the expression of the MMP enzyme, thereby inhibiting skin elasticity degradation and wrinkle formation.

Hereinafter, although other formulation examples of the present invention are exemplified, the formulation of the cosmetic composition according to the present invention should not be construed as being limited thereto, and conventional variations of those skilled in the art are possible within the scope of the present invention.

Formulation Example 1. Softening Longevity number ingredient content(%) One glycerin 5.00 2 1,3-butylene glycol 3.00 3 PEG 1500 1.00 4 Allantoin 0.10 5 DL-panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 ethanol 10.00 9 Octicdodeceth-16 0.20 10 Polysorbate 20 0.20 11 Sample 3 5.0 12 Preservative, fragrance, coloring a very small amount 13 Distilled water Remaining amount

Formulation Example 2. Converging Cosmetic Water number ingredient content(%) One glycerin 2.00 2 1,3-butylene glycol 2.00 3 Allantoin 0.10 4 DL-panthenol 0.30 5 EDTA-2Na 0.02 6 Benzophenone-9 0.04 7 ethanol 15.00 8 Polysorbate 20 0.20 9 Sample 3 1.0 10 Citric acid a very small amount 11 Preservative, fragrance, coloring a very small amount 12 Distilled water Remaining amount

Formulation Example 3. Nutrients number ingredient content(%) One Cetearyl alcohol 1.00 2 Glyceryl Stearate 0.50 3 Polysorbate 60 1.00 4 Sorbitan sesquioleate 0.30 5 Cetyloctanoate 6.00 6 Squalane 4.00 7 Flower oil 4.00 8 Butylene Glycol 4.00 9 glycerin 4.00 10 Carbomer 0.10 11 Triethanolamine 0.10 12 Sample 3 1.00 13 Preservative, fragrance, coloring a very small amount 14 Distilled water Remaining amount

Formulation Example 4. Essence number ingredient content(%) One glycerin 10.00 2 Betaine 5.00 3 PEG 1500 2.00 4 Allantoin 0.10 5 DL-panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 Hydroxyethyl cellulose 0.10 9 Carboxyvinyl Polymer 0.20 10 Triethanolamine 0.18 11 Octyldodecanol 0.30 12 Octyldodeceth-16 0.40 13 ethanol 6.00 14 Sample 3 1.00 15 Preservative, fragrance, coloring a very small amount 16 Distilled water Remaining amount

Formulation Example 5 Pack number ingredient content(%) One Polyvinyl alcohol 15.00 2 Cellulose gum 0.15 3 glycerin 3.00 4 PEG 1500 2.00 5 Chicdestrin 0.15 6 DL-panthenol 0.30 7 Allantoin 0.10 8 Glycyrisin Monoammonium 0.20 9 Nicotine amide 0.40 10 ethanol 5.00 11 PEG 40 Cured Castor Oil 0.30 12 Sample 3 1.00 13 Preservative, fragrance, coloring a very small amount 14 Distilled water Remaining amount

Claims (5)

As a cosmetic composition for antioxidant, wrinkle improvement and skin elasticity improvement,
A cosmetic composition comprising a phytoPlacenta complex extract of beans, lotus, peas, magnolia, camellia, plum and peony in a weight ratio of 3: 2: 1: 1: 1: 1: 1. The method of claim 1,
The composite extract is a cosmetic composition, characterized in that extracted with ethanol. The method of claim 1,
The complex extract is a cosmetic composition, characterized in that contained in an amount of 0.0005 to 10% by weight based on the total weight of the composition. delete delete

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