KR20180010765A - Cosmetic composition comprising the extract of ishige okamurae - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin elasticity in skin, emulsion, essence or cream formulations by comprising an extract of Ishige okamurae which is a marine plant in brown algae. The cosmetic composition of the present invention has excellent texture, has no stimulation and can improve skin elasticity.

Description

[0001] The present invention relates to a cosmetic composition comprising an extract of rosemary extract,

The present invention relates to an extract of Ishige okamurae, a marine plant belonging to the group of brown algae, and is useful for improving the skin elasticity of skin, emulsion, essence or cream formulations, improving dermal elasticity, improving elastic restoring force, improving skin pore, To a cosmetic composition having an effect of improving skin texture.

The phenomenon of 'well-being', which has a great influence on modern society, includes not only the primary meaning of 'eating well and living well', but also the meaning of 'the health of the body is expressed by the beauty of skin'. This phenomenon is also seen in the field of skin care, and the approach to skin care now requires a complex function and a scientific approach, rather than seeking a piecemeal efficacy (Kim, Eun Young, Korean Society of Skin Care, 2005).

Skin aging can be roughly classified into two types. One of them is intrinsic aging, which is an inevitable aging phenomenon as time goes by. The second is photoaging, which refers to the aging phenomenon observed in the skin such as the face, the back of the hand, and the back of the neck exposed to sunlight for a long time, resulting from the combination of the effects of endogenous aging and ultraviolet rays (Kim, JG Sci Nutr, 2010).

The pores are distributed throughout the face skin, but the nose and the ball area can be distinguished by the naked eye. There are individual differences in the appearance of the pores due to the endogenous and exogenous factors such as sex, genetic, aging, acne and exposure to chronic ultraviolet rays. The reason for the widening of the pores is that as the sebaceous glands are excessively secreted or the aging of the skin begins, collagen fibers and elastic fibers supporting the pore walls are denatured or decreased, and the skin elasticity is lost and stained Society, 2010).

It is known that the cause of skin aging is a collapse of matrix proteins such as collagen and elastic fibers, resulting in deficiency of collagen in the skin and denaturation of elastic fibers. In addition, during the progress of skin aging, collagen synthesis is reduced through various signal transduction pathways, and expression of matrix metalloproteinases (MMPs), which are degradation enzymes of extracellular matrix proteins including collagen, is increased It was announced. As such, collagen has been recognized as a major factor related to skin aging, and inhibition of collagen synthesis and inhibition has been an important index for skin aging inhibition and skin care.

Various studies have been conducted to improve collagen synthesis and to inhibit its degradation to improve skin beauty. Natural products having excellent skin stability are attracting interest in comparison with compounds.

Korean Patent No. 10-100898758 (entitled " Extract of Lactobacillus plants showing antidiabetic activity)

An object of the present invention is to provide a cosmetic composition in the form of a skin, an emulsion, an essence or a cream containing a decolorized or deodorized cordierite extract as an active ingredient.

Hereinafter, this specification will be described in detail.

Whenever a component is referred to as "comprising ", it is understood that it may include other components as well, without departing from the other components unless specifically stated otherwise.

The present invention provides a cosmetic composition comprising a decolorized or deodorized Lard extract and in the form of a skin, emulsion, essence or cream.

In this specification, the beard is a plant belonging to brown algae among marine plants, and is known to contain various minerals and vitamins. The plaque contains a large amount of diphloretho-hydroxycarmalol (DPHC), a kind of phlorotannin, which acts as an index component of the extract of Lach.

In the present specification, decolorization refers to the absorption or decomposition of contaminating colors or pigments of a material. In this specification, deodorization may mean the removal of volatile substances, which are unpleasant to the olfactory sense.

The decolorization or deodorization of the present invention can be carried out using activated carbon, but it is not limited thereto as long as it can decolorize or deodorize and does not affect the extract of Lach. However, the use of activated carbon is more preferable in view of not being affected by diphloretho-hydroxycarmalol (DPHC), which is a kind of phlorotannin.

The present invention can provide a cosmetic composition which is excellent in feeling when used as a pollutant contained in a Lactuca extract or commercialized by containing a decolorized or deodorized Lactuca extract. In the present specification, it is more preferable to include the decolorized and deodorized Cord extract.

The composition of the present invention can be applied by a conventional method of applying a cosmetic composition according to purposes. Preferably, the composition can be directly applied to the skin as an external preparation for skin and then transdermally applied. When applied to the skin, the treated area may be the entire skin surface of the subject or only the required area. Application of the composition may be repeated periodically.

In the present invention, the Linalool extract can be obtained by a conventional method known in the art, for example, using a solvent extraction method using a conventional solvent under the conditions of ordinary temperature and pressure. The extraction process may be carried out by extraction with about 2 to 20 times, preferably about 5 to 10 times, the primary extraction solvent with respect to the weight of the tablet. Extraction can be carried out by methods known in the art such as, for example, cold-pressing, hot water extraction, ultrasonic extraction, reflux-cooling extraction, and the like. The extraction temperature can be varied by a person skilled in the art in a variety of temperature ranges suitable for the extraction method, for example, but not limited to, from 20 캜 to 100 캜. In addition, the extraction time differs depending on the extraction method, and a person skilled in the art can adopt an appropriate extraction time, and can be performed in a single or multiple cycles within a range of about 1 hour to several days, though not limited thereto. The extract obtained by performing the extraction with the primary extraction solvent may be obtained in the form of a liquid in which impurities are removed by filtration according to a conventional method, or the obtained liquid form extract may be concentrated under reduced pressure and / Can be obtained.

As an extraction solvent preferred in the present invention, ethanol, more preferably aqueous 30% to 70% (v / v) ethanol solution may be used. More preferably, an aqueous 50% (v / v) ethanol solution can be used.

The extraction step may further include, if necessary, obtaining a fraction having a high active ingredient content. That is, the step of dispersing the extract obtained by extracting with the primary extraction solvent into water and then extracting with an appropriate secondary extraction solvent, for example, alcohol of water-saturated C4, is performed to further increase the active ingredient content in the resulting extract .

In addition, the Lactobacillus extract of the present invention may contain not only the extract obtained by the above-mentioned extraction solvent, but also an ordinary purified and fermented extract. For example, decompression by carbon dioxide, extraction by supercritical extraction with high temperature, extraction by extraction using ultrasound, separation by ultrafiltration membrane with constant molecular weight cutoff value, various chromatography (size, charge, hydrophobic or affinity And the active fraction obtained through various purification and extraction methods, such as those extracted by fermentation products using natural or various microorganisms, are also included in the extract of the present invention.

The present invention may comprise 0.1 to 10% by weight of the decolorized or deodorized cord extract on the basis of the total weight of the cosmetic composition. In the present invention, it is more preferable that the skin care composition contains 0.1% by weight to 1% by weight of the decolorized or deodorized cord extract on the basis of the total weight of the cosmetic composition in terms of skin improving efficacy and / or stability.

In the case of a composition containing the composition ratio, it corresponds to a composition ratio found as a result of a study on composition ratios showing excellent skin elasticity improvement effect without skin irritation. However, the content is not limited thereto.

The decolorized or deodorized plaque extract of the present invention may further comprise a high pressure dispersion treatment process.

In the present specification, it is possible to minimize the physiological activity by stabilizing and absorbing the skin of the Lactuca extract, which is an active ingredient contained in the cosmetic composition, through the high-pressure dispersion treatment.

In the present invention, the high-pressure dispersion treatment is preferably performed using a high-pressure homogenizer (microfluidizer). The present inventors have confirmed in the examples described below that the Cordyceps extract can provide a fine particle size when a high pressure homogenizer is used.

The high-pressure homogenizer refers to a device for finely atomizing particles by applying impact force, shearing force, or cavitation to the particles in the fluid by using a velocity generated by passing the fluid through the micro dispersion device.

In the present invention, the high-pressure dispersion treatment is preferably carried out using 1,3-butanediol. When 1,3-butanediol is used as a dispersion solvent in the high-pressure dispersion treatment process, it is possible to provide a fine particle size.

In the present specification, the cosmetic composition has the effect of improving skin elasticity, improving dermal elasticity, improving elastic restoring force, improving skin pore, improving dermal density, or improving skin texture.

The method for measuring skin elasticity, dermal elasticity, elastic restoring force, skin pore, dermal density, skin texture improvement can be measured by a method described later, but is not limited thereto.

In the present specification, the term 'cosmetic material' refers to a substance which can be a cosmetic or a quasi-drug.

Cosmetics refers to products that are used in the human body to maintain or improve the health of skin and hair by adding cleanliness and beauty to the body, changing appearance, brightening the appearance, and acting on the human body. Functional cosmetics are products that help to whiten the skin, help to improve the wrinkles of the skin, protect the skin from ultraviolet rays, and include those prescribed by the Ministry of Health, Welfare and Family Affairs. do.

Quasi-drugs refer to medicines classified by the Ministry of Health and Welfare for products that have a slight effect on the human body, rather than medicines used to treat or prevent diseases. According to the Pharmaceutical Affairs Law, textile or rubber products used for the treatment or prevention of diseases of humans or animals, excluding those used for medicinal purposes, , Sterilization and insecticides to prevent infectious diseases, and the like. Typical examples of quasi-drugs include toothpaste, functional soap, and functional shampoo.

The cosmetic composition of the present invention has formulations of skins, emulsions, essences or creams. In addition, the composition of the present invention may be prepared in any formulations conventionally produced in the art, and may be in the form of, for example, emulsion, skin, essence, cream, eye cream, pack, gel, powder, lipstick, The composition may be a formulation selected from lotions, ointments, patches, serum, cleansing foam, cleansing cream, cleansing water, body lotion, body cream, body oil, body essence, shampoo, rinse, body cleanser, soap and spray.

The cosmetic composition of the present invention, in addition to the above-mentioned components, may be blended with other components, which are usually added to cosmetics as required. For example, it is possible to use water, water, a moisturizing agent, a sequestering agent, a pH adjusting agent, a surfactant, a skin softening agent, a moisture blocking agent, an antioxidant, a conditioning agent, a sterilizing preservative, , One or more additives selected from the group consisting of alcohol, solubilizing agent, perfume, oil, synthetic high molecular substance, neutralizing agent, natural extract, pigment, pigment, stabilizer, vitamin and chelating agent.

The additive may comprise conventional materials used in the art. For example, the cosmetic composition of the present invention can be used in cosmetics, cosmetics, cosmetics, cosmetics, cosmetics, pharmaceuticals, cosmetics, pharmaceuticals, Aloe vera leaf extract, acetyloctapeptide-3, 1,2, -hexanediol, phenyl triethoxysilane, phenylglycine, phenylglycine, ethylhexanoate, hydrogenated vegetable oil, tocopheryl acetate, sunflower seed oil, fragrance, Hydrogenated castor oil, ornithine, salicylic acid, clophenecin, panthenol, glycereth-26, sodium hyaluronate, ethanol, tamarind seed polysaccharide, butylene glycol But are not limited to, carrageenan, corn, inulin, cetearyl alcohol, cyclopentasiloxane, dimethicone, meadowfoam seed oil, caprylyl glycoll, acetyloctapeptide-4, glycolipid and caprylic / capric triglyceride Of one or more additives selected from the group consisting of Jethro it may further include.

 The preparation method of these formulations can be produced according to a conventional method of producing a cosmetic formulation.

The composition of the present invention may also be applied in an amount effective to the skin of a human or other animal to improve skin elasticity.

As used herein, "effective amount" means the amount of active ingredient or composition that elicits a biological or medical response in an animal or human considered by a researcher, veterinarian, physician or other clinician. It will be apparent to those skilled in the art that the duration and frequency of application of the cosmetic composition of the present invention will vary with the desired effect.

The cosmetic composition of the present invention is excellent in feeling of use and can significantly improve skin elasticity without stimulation.

Fig. 1 is a photograph taken before and after decolorization of the extract of Lard using activated carbon.
FIG. 2 (A) is a graph showing the DPHC content of the extract obtained by extracting 1 kg of the glandular material with 50% alcohol, by HPLC, and FIG. 2B is a graph showing the DPHC content as an index material when the extract is decolorized with activated carbon It was confirmed by HPLC.
Fig. 3 is a schematic view showing a method for producing a Lycopersicon esculentum extract.
4 is a photograph (right side) and a working principle (left side) of a high pressure homogenizer used in the production of the present invention.
5 is a schematic view showing the process of measuring the content of the indicator material and the particle size after the preparation of the extract of the present invention and the high pressure dispersion treatment.
FIG. 6 is an HPLC chart showing the indicator material (DPHC) content of the extract of the present invention after treatment with a high-speed homogenizer or a high-pressure homogenizer.
FIG. 7 is a view showing particle size distribution after treatment with a high-speed homogenizer or a high-pressure homogenizer.
FIG. 8 is a graph showing the indicator material (DPHC) content of the Lactobacillus sp. Extract when the dispersion solvent is differently dispersed under high pressure. Cond. 1 is distilled water (DW), Cond. 2 is ethanol (EtOH), and Cond. 3 is 1,3-butanediol as a dispersion solvent.
FIG. 9 is a graph showing the particle size distribution when the dispersing solvent is changed and dispersed under high pressure. 9 (a) shows distilled water, FIG. 9 (b) shows ethanol, and FIG. 9 (c) shows a case where 1,3-butanediol is used as a dispersion solvent.
10 is a graph showing the results of confirming cytotoxicity against human skin fibroblasts.
Fig. 11 shows the result of confirming the expression of Pro-Collagen gene by Competetive RT-PCR.
Fig. 12 shows the results of confirming the expression of Pro-Collagen gene by qPCR.
Fig. 13 shows the result of confirming the expression of Elastase gene by qPCR.
14 is a photograph of a 15% crude extract (pre-M / F) (left) and a 15% crude extract (post-M / F) (right).
Fig. 15 is a graph showing changes in skin elasticity of a test subject when four kinds of cosmetic compositions (skins, emulsions, essences, creams) containing the extract of the present invention are used.
FIG. 16 is a graph showing change in dermal elasticity of a test subject when four kinds of cosmetic compositions (skins, emulsions, essences, creams) containing the extract of the present invention are used.
Fig. 17 is a graph showing changes in elastic restoring force of a test subject when four kinds of cosmetic compositions (skins, emulsions, essences, creams) containing the extract of the present invention are used.
18 is a graph showing changes in skin pores of a subject to be tested when four types of cosmetic compositions (skins, emulsions, essences, creams) containing the extract of the present invention are used.
FIG. 19 is a graph showing changes in dermal density of a subject to be tested when four kinds of cosmetic compositions (skins, emulsions, essences, creams) containing the extract of the present invention are used.
20 is a graph showing changes in skin texture (skin roughness) of a test subject when four kinds of cosmetic compositions (skins, emulsions, essences, creams) containing the extract of the present invention are used.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

Example  1. Method for manufacturing Linalool extract

A plaque produced in May and July was supplied from the fishing village of Ayeol-eup in Jeju Island and washed and prepared.

(1) Establishment of drying conditions

100 g of the washed pellets were placed in a freeze dryer (reflow) and dried at a low temperature of vacuum (-50 ° C) for 12 to 15 hours. Further, 100 g of the washed pellets were placed in a high-temperature drier (Vision Science, PL-S11) and dried at 60 DEG C for 12 to 15 hours. After drying, weights were measured and found to be 97-98 g in both cases. Thus, it was confirmed that both of the above two drying conditions were suitable for use in the preparation of the Lactobacillus extract of the present invention.

(2) crushing and extracting

50 g of the lyophilized cake was placed in a grinder (super grinder JL-1000) and repeatedly turned on / off five times at intervals of 30 seconds / 10 seconds. After grinding, Were weighed and weighed. 100 ml of alcohol alcohol was added to 10 g of the pulverized powder, and the mixture was extracted at 50 캜 for 2 hours.

(3) Color and deodorization

The activated charcoal was separated by centrifugation at 8,000 rpm for 10 minutes, and the supernatant was filtered with a filter paper. The filtrate was filtered through filter paper And the activated carbon was completely removed.

As a result, the odor of the extract was reduced due to the removal of the fishy odor which is characteristic of the extract of Lard. It was also confirmed that the color of the Lactobacillus extract was changed from turbid to clear brown (Fig. 1).

In addition, before and after the decolorization and deodorization with activated carbon, the cord extract was subjected to HPLC analysis under the following conditions.

- Detector: Waters 996

- solvent conditions

A: DW + 0.1% TFA

B: ACN + 0.1% TFA

- Injection volume: 20 μl

- Detector wavelength: 230 nm

- use column: UG80 5 ㎛ size

(Shiseido / C18 / 4.6 mm.D x 250 mm / Japan)

- flow rate: 0.7 ml / min

The HPLC results are shown in Table 1 and FIG. 2 below.

[Table 1]

In Table 1 and FIG. 2, when the area ratio of DPHC peak as an index substance of the extract of Lach is compared with the result of HPLC analysis, the DPHC content after the final decolorization (61,599,658) (FIG. 2B) after the 50% alcohol alcohol extraction (47,601,731) ) Was increased about 1.3 times. In other words, the indicator material content of the Lactobacillus extract is hardly affected by the decolorization and deodorization process using activated carbon.

(4) Process for manufacturing Linalool extract

Fig. 3 is a schematic diagram of the process for producing the Lactobacillus extract of the present invention. The prepared lobes were washed, dried and pulverized, extracted, centrifuged, filtered and concentrated. The lyophilized lyophilized product was referred to as a second lyophilized product 3). The second lyophilized extract is decolorized and deodorized with activated charcoal, and then concentrated. The lyophilized lyophilized product is referred to as a third lyophilized product (the first, second and third Process of heat). In the following, in vitro experiments were performed using a second lyophilized product, and each formulation and experimental example used a third lyophilized product.

Example 2. Application of high pressure dispersion process to rosemary extract

The high-pressure dispersion process requires initial investment costs, but is advantageous in terms of long-term production management. To investigate the feasibility of applying the high pressure dispersion process to the extract of rosemary to make cosmetics containing rosin extract using nano emulsification system,

Specifically, experiments were conducted mainly on the application of high speed homogenizer and high pressure homogenizer, which are typically used as mechanical nanoemulsification and high pressure dispersion. The high-speed homogenizer is generally similar to the ultrasonic method, and it is easy to crush and disperse the sample by the force by cavitation and the mechanical shear force. Typical high-speed homogenisers are blenders that are usefully used for sampling in plant and animal tissues by cutting blades mounted above or below. In general, the high pressure homogenizer is a dispersion and emulsification system having a narrowest particle size distribution under a constant pressure condition.

The operation principle of the high-pressure homogenizer (ISA-N-30A, Ilshin Autoclave Co., Ltd., Korea) used in the present invention (see FIG. 4) is as follows. Ultrahigh pressure generated in the intensifier pump installed in the high pressure homogenizer is applied to the sample, It moves through the pipe to the interaction chamber and passes through the micro-channel inside the chamber, causing nano dispersion and nano emulsification. The pressure of the sample or target components is not released immediately but is transferred through the micro-pipe and the micro-channel. Unlike high-speed homogenizer, homogenizer of high pressure homogenizer is different from homogenizer of high-pressure homogenizer because pressure constantly changes in one pressure stroke. Therefore, particle size and particle size distribution in one pressure stroke The particle size distribution is very uniform when the pressure applied to the sample per pressure stroke is kept constant.

(1) High pressure dispersion process condition search

The mechanical high pressure dispersion method suitable for the present invention was investigated, and various high pressure dispersion conditions were investigated for the final high pressure dispersion process to establish optimal conditions for high pressure dispersion.

Lactobacillus extract was prepared in the same manner as in Example 1, and used in this experiment while refrigerated.

A schematic diagram of the experimental procedure is shown in FIG.

The HPLC analysis conditions for the indicator component (DPHC) of the Lactobacillus root extract are shown in Table 2 below.

[Table 2]

First, the application of L - extracts to high - speed homogenizer and high - pressure homogenizer was investigated by mechanical high - pressure dispersion method. The samples were diluted 10-fold with 10% ethanol extracts against 10% extracts and used in this experiment. The high-speed homogenizer was an Ultra Turrax (T18) from IKA (Germany) and was treated at 5,000 rpm for 30 min. The high-pressure homogenizer was ISA-N-30A from Ilshin autoclave. The pressure of the high-pressure dispersion was controlled to 1,000 bar (14,500 psi) and the micro-channel or nanocell was maintained at 4 ° C for 30 min. The control (control) was a non-treatment condition without high pressure dispersion treatment. All the experiments were carried out while thoroughly washing with distilled water, methanol or the like in consideration of the internal volume and the like in order to prevent cross contamination by the condition. The conditions for setting the high-speed homogenizer and the high-pressure homogenizer are as follows.

[Table 3]

Results confirm the content change of the indicator component DPHC according to the diluting solvent of ethanol using a high pressure dispersion for L extract (Fig. 6), the untreated (control), a high speed homogenizer, t _R 10.6 of the HPLC chromatograms from high-pressure homogenizer treatment conditions (DPHC) as an indicator component of the extracts of high-pressure liquid was increased according to the order of high-pressure homogenizer> high-speed homogenizer> control in the peak area of 239,572, 265,475 and 285,846 respectively Especially, the high pressure homogenizer was increased to 119% compared to the non - treatment condition, and it was judged that the high pressure homogenizer process was suitable for the high pressure dispersion process.

As a result of particle size analysis, the particle size of 15.08, 14.44, and 14.35 μm was found to be 15.08, 14.44 and 14.35 μm, respectively, in the control, high-speed homogenizer and high-pressure homogenizer (Fig. 7) It was confirmed that the particle size was smaller in the mechanical high pressure dispersion than in the untreated water.

The results of DPHC and particle size change according to high pressure dispersion conditions for Lahoo extract are shown in Table 4 below.

[Table 4]

(2) High pressure homogenizer  Distributed solvent review for application

As a result of application to rope extract for high pressure homogenizer process, it is possible to select the optimal dispersion solvent which is one of the important factors in application of high pressure homogenizer, distilled water which can be used as base material of cosmetics, ethanol and 1,3 -butanediol as a dispersing solvent for the high pressure homogenizer for Lard extract as shown in Table 5 below.

[Table 5]

As the evaluation method, the high pressure homogenization process was applied to each dispersed solvent in the same manner and the DPHC content, which is an indicator active ingredient of the extract of Lard, was compared and analyzed. As shown in the graph of FIG. 8, the HPLC analysis showed that the profiling of the chromatograms for each condition was the same, and the results of the DPHC content by each condition were as follows: 1,3-butanediol>EtOH> DW , 1,716 × 10 ³ , 758 × 10 ³ , and 288 × 10 ³ , respectively, and 1,3-butanediol was selected as the optimal dispersion solvent for the high pressure homogenizer of the extract. 1,3-butanediol is also considered to be suitable for application to the cosmetic composition of the present invention because it has the effect of decreasing viscosity, improving dry or damaged skin, improving flexibility, and the like as a skin conditioner.

The results of particle size analysis (FIG. 9 and Table 6) for the application of the high pressure homogenizer to each disperse solvent condition were 0.64, 14.35, and 0.02 um in DW, EtOH and 1,3-butanediol, And the size of the particles of 1,3-butanediol <DW And EtOH in the order of 1, 3-butanediol. In the case of EtOH, the solubility of the extract was lower than that of DW and 1,3-butanediol. It is determined that the particle size is relatively large regardless of the particle size.

[Table 6]

Example 3. Evaluation of cytotoxicity and skin elasticity-related genes of Lactobacillus root extract

(1) Evaluation of cytotoxicity

15% Lactobacillus root extract was prepared and suspended in HS27 cells (Human Skin Fibroblast cell) at a concentration of 0, 0.01, 0.1, 0.5 or 1 mg / ml before microemulsion treatment (pre-MF) Respectively.

In addition, lyophilized 100% - Lilac extract and lyophilized 100% - Lilac extract were applied to HS27 cells in the same manner.

The results are shown in FIG. In FIG. 10, a, b, and c mean significant differences, and P <0.05 (the same in the following drawings). 10 showed no cytotoxicity up to the concentration of 0.1 mg / ml in both the pre-MF and post-MF of the second-tall roe extract in the form of solution, and in the lower two stages, the powdery second- And the third lyophilized lyophilisate showed no cytotoxicity up to the concentration of 0.01 mg / ml.

(2) Evaluation of effect on pro-collagen gene expression

Increased collagen is known to increase skin elasticity. Therefore, the effect of lyophilized pre-MF and post-MF, freeze-dried second-tall extract of lard extract and lyophilized third-tall extract on the expression of COLA1 gene, a Pro-Collagen gene, was confirmed by competitive-RT-PCR and qPCR Respectively.

The results of Competetive RT-PCR are shown in the left side of FIG. 11, and are shown graphically on the right side. In the left and right diagrams of FIG. 11, the expression of the COLA1 gene was significantly increased by the pre-MF and post-MF extracts of the second-tall cell extracts, and the expression of COLA1 gene was significantly increased by the third- The freeze - dried extract showed no significant effect compared with the control group.

The qPCR results are shown in Fig. 12, the same results as in the case of Competetive RT-PCR were obtained in the case of qPCR.

(3) Evaluation of effects on elastase gene expression

It is known that when elasticity increases, skin elasticity increases, and when elasticity is broken down by elastase, skin elasticity decreases. Therefore, qPCR was used to confirm the effect of lyophilization of pre-MF and post-MF, lyophilized lyophilized powder extract and lyophilized third-generation LH extract on the expression of Elastase gene, a Pro-Collagen gene.

The results are shown in Fig. In FIG. 13, the post-MF of the second tannin extract significantly reduced Elastase gene expression. However, other samples showed no significant effect.

(4) Theorem

The results of the above tests (1) to (3) are summarized as follows. As a result, it was confirmed that the ingredients of the cosmetic composition for improving the skin elasticity in the order of the second-tannin extract, the pre- Respectively.

Photographs of 15% crude extract (pre-M / F) and 15% crude extract (post-M / F) are shown on the left and right sides of FIG.

Example 4. Development of Skin Elastic Cosmetic Formulation Containing Lard Extract

(1) Emulsion Formulation

1) Preparation of emulsion formulations

An emulsion formulation having the composition shown in the following table containing 0.1%, 0.5%, or 1.0% of the Linalool extract prepared in Example 1 was prepared. The manufacturing method is also shown in the right column of the table below.

[Table 7]

2) Stability test of emulsion formulations

The prepared emulsion was added to the emulsion under the conditions of 0 ° C, room temperature (25 ° C), 45 ° C, 50 ° C, or cycling (extreme conditions: 4, 0, 25, 45, After storage for 1 ~ 7 days, after 8 ~ 14 days storage, after 15 ~ 21 storage, for 22 ~ After storage for 30 days, the stability was checked by confirming the stability of the formulation, discoloration and detachment, etc. The stability of the emulsion formulation containing 1.0% of the extract of Lard was found to be the best.

[Table 8]

(2) Skin Formulation

1) Preparation of skin formulations

A skin formulation having the composition of the following table containing 0.1%, 0.5%, or 1.0% of the Linalool extract prepared in Example 1 was prepared. The manufacturing method is also shown in the right column of the table below.

[Table 9]

2) Stability test of skin formulations

The stability of the prepared skin formulations was tested in the same manner as in (1) above, and as a result, there was no problem in stability under all conditions.

The physical properties (pH) of the skin formulations containing 0.1%, 0.5% or 1.0% of the extract of Lard were found to be pH 5.05, 5.07 and 5.06, respectively. As a result, it was found that the effect of Lard extract on the pH of the skin formulations was insignificant I could.

(3) Formulations of cream

1) Manufacture of cream formulations

A cream formulation having the composition of the following table was prepared. The manufacturing method is also shown in the right column of the table below.

[Table 10]

2) Stability test of cream formulations

A cream formulation containing 0.1%, 0.5%, or 1.0% of the Lard extract prepared in Example 1 was prepared. The stability of the prepared cream formulation was tested in the same manner as in (1) above. As a result, it was found that the stability of the emulsion formulation containing 1.0% of Lard extract was the best in all conditions.

As a result of investigation of the physical properties (pH and viscosity) of the cream formulations containing 0.1%, 0.5%, or 1.0% of the extracts of rosemary extracts, it was found that the effect of the roots extract on the pH and viscosity of the cream formulations was insignificant there was.

[Table 11]

(4) Essence formulation

1) Preparation of essence formulation

An essence formulation having the composition of the following table containing 0.1%, 0.5%, or 1.0% of the Lard extracts prepared in Example 1 was prepared. The manufacturing method is also shown in the right column of the table below.

[Table 12]

2) Stability test of the essence formulation

The stability of the prepared essence formulation was tested in the same manner as in (1) above. As a result, the stability of the essence formulation containing 1.0% of the extract was found to be the best in all conditions.

The physical properties (pH and viscosity) of the essence formulation containing 0.1%, 0.5%, or 1.0% of the extract of Lard were respectively examined. As a result, it was found that the effect of Lard extract on the pH and viscosity of the essence formulation was insignificant there was.

[Table 13]

Test Example 1. Skin elasticity improvement test

Four types of skin elastic cosmetic formulations (skins, emulsions, essences, creams) containing the 1.0% extract prepared in Example 4 were subjected to the following clinical tests.

(1) Test subject and test method

At the KC Skin Clinical Research Center, a total of 21 adult women aged 19-59 years were tested. The average age of subjects was 43.429 ± 6.630 years.

The subjects were asked to visit the center three times in total. At the first visit, subjects were selected by obtaining the consent of the subjects and confirming the exclusion criteria. After cleansing with a cleanser prepared before use of the test product, the skin is stabilized in a space of constant temperature and humidity condition (temperature 20 ~ 24 ℃, humidity 40 ~ 60%) for about 30 minutes, The site to be applied was photographed or measured. The test product and logbook were then issued.

The test subjects were allowed to use the test product by cleansing the entire face twice a day, followed by spreading an appropriate amount of the skin-essence-emulsion-cream in order.

 After 2 weeks of using the test product, the subject is allowed to visit the center for the second time, and the skin is stabilized for about 30 minutes in a space of constant temperature and humidity condition (temperature 20 to 24 ° C, humidity 40 to 60% And adapted to the measurement environment, and then the test product application site was photographed or measured. In addition, the adverse reaction was confirmed and the subjects' questionnaire was evaluated. After 4 weeks of using the test product, the test subject is made to visit the center for the third time, and the same researcher shoots or measures the test product application area in the same way as the second visit, confirms the adverse reaction, Respectively.

(2) Skin efficacy evaluation item

1) Skin elasticity measurement

Skin elasticity was measured by using a cutometer, and skin elasticity was measured before, 2, and 4 weeks after the test product. The R2 value was used as the skin elasticity evaluation data. The measured skin elasticity unit is A.U (Arbitrary Unit), and the measured value is proportional to the skin elasticity.

2) Dermal elasticity measurement

Dermal elasticity was measured by using dermal torque, and skin elasticity was measured before, 2, and 4 weeks after the test product. Ur / Ue values were used as skin dermal elasticity evaluation data. The measured skin elasticity unit is A.U (Arbitrary Unit), and the measured value is proportional to the skin elasticity.

3) Elastic restoring force measurement

Elastic restorative force was measured by ballistometer before and after 2 weeks and 4 weeks. The average value measured three times was used as elastic restoring force evaluation data. The measured elastic restitution force is a CoR (Coefficient of Restitution) value, and the measured value is proportional to the degree of improvement in elastic restoring force.

4) Skin pore measurement

Using the Janus 1 Mark II, the entire face was taken before, 2 weeks after, and 4 weeks after using the test product. The area of the pores of the photographed images was analyzed to analyze the degree of skin pore and used as evaluation data. The measured skin pore unit is%, and the average pore size of the analyzed area appears in 100% of the total, and the% value and the skin pore degree are in inverse proportion, and the smaller the measured value, the better the pore improvement.

5) Measurement of dermal density

Dermal density was measured by using Skin Scanner USB. The dermal density of the dorsal surface of the face was measured before, 2, and 4 weeks after the test product was used. The density of the dermis of the measured image was analyzed and used as the dermal density. The unit of dermal density is%, and the measured value is proportional to the improvement of dermal density.

6) Measurement of skin texture

Skin texture measurements were taken using Antera 3D and the skin roughness was measured before, during, 2 weeks and 4 weeks after the test product. The degree of roughness was analyzed by specifying specific parts of the photographed photographs and used as skin texture evaluation data. The measured roughness parameter is Ra, and the analytical value and the degree of roughness are in inverse proportion, which means that the lower the analysis value, the better the skin texture.

(3) Evaluation of skin adverse reaction

To ensure the safety of the subjects, surveys and visual evaluations of adverse events were conducted at each visit, and the occurrence and symptom severity were checked. Adverse reactions and adverse events were determined in appropriate measures and participation of subjects in the test.

(4) Assessment of compliance with test product use

The test subjects were instructed to use the product when using the test product according to the prescribed usage and frequency of use, and to write it in the compliance logbook distributed with the test product. The test subjects who used less than 80% of the 100% compliance level of the test product should be excluded from the analysis of the measurement data.

(5) Evaluation of Questionnaire

The subjects were asked to directly evaluate the effectiveness and likelihood of skin improvement after using the test product at each visit with a total of 5 steps (very satisfied ~ very unsatisfactory).

(6) Method of statistical analysis

SPSS 23.0 was used for statistical analysis to determine the significance of changes in skin measurements before and after the test product use. Statistical significance was confirmed at p <0.05 at 95% confidence interval.

1) Comparison before and after test product use (single measurement)

- Normalized test satisfaction: Paired samples T-test (parametric method)

- Normality test dissatisfaction: Wilcoxon signed ranks test (nonparametric method)

2) Comparison of test products after n hours (multiple measurements)

- Normality test satisfaction: One-way ANOVA (parametric method)

- Normality test Dissatisfied: Kruskal-wallis test (non-parametric method)

(7) Test results

A total of 21 subjects were dropped from the test, and a total of 20 subjects completed the test. The average compliance of the subjects with the product use was 98.928%.

Skin efficacy evaluation and anomalous evaluation results are as follows.

1) Skin elasticity measurement result

[Table 14]

As shown in Table 14 and FIG. 15, skin elasticity values were significantly increased after 2 weeks and 4 weeks of use of the test product compared to before use of the product (p <0.05).

2) Results of dermal elasticity measurement

[Table 15]

As shown in Table 15 and FIG. 16, after 2 weeks and 4 weeks of using the test product, the dermal elasticity value was significantly increased compared to before use (p <0.05)

3) Elastic restoring force measurement result

[Table 16]

As shown in Table 16 and FIG. 17, after 2 weeks and 4 weeks of using the test product, the elastic restoring force was statistically significantly increased compared to before use (p <0.05)

4) Results of skin pore measurement

[Table 17]

As shown in Table 17 and FIG. 18, after two weeks and four weeks of using the test product, the skin pores were significantly decreased compared to before use (p <0.05)

5) Results of dermal density measurement

[Table 18]

As shown in Table 18 and FIG. 19, after 2 weeks and 4 weeks of use of the test product, the dermis density was significantly higher than that of the product before use (p <0.05)

6) Skin texture measurement result

[Table 19]

As shown in Table 19 and FIG. 20, after 2 weeks and 4 weeks of using the test product, the skin roughness was significantly lower than that of the product before use (p <0.05)

7) Test subjects' abnormal reaction evaluation results

None of the subjects reported skin adverse events (erythema, edema, stinging, itching, irritability, burning sensation, stiffness, tingling) when evaluated 2 weeks and 4 weeks after use of the product.

8) Results of the questionnaire survey

Table 20 shows the results of the questionnaire evaluation after 2 weeks of using the product, and Table 21 shows the results of the questionnaire evaluation after 4 weeks of using the product.

[Table 20]

[Table 21]

As shown in the above Tables 20 and 21, the score of each item and the overall satisfaction score of the test subjects were 2 or more weeks after using the product. 4 points) or more.

Test example 2. Skin irritation test

(1) Test subject and test method

A patch was prepared by dropping 20 μl of each test substance (skin, emulsion, essence, tension cream) into the IQ chamber. The following tests were conducted using this patch.

At the KC Skin Clinical Research Center, a total of 32 adult women aged 19 to 59 years were tested. The average age of subjects was 42.188 ± 7.876 years.

The subjects were asked to visit the center three times in total. At the first visit, subjects were selected by obtaining the consent of the subjects and confirming the exclusion criteria. The subjects were allowed to adapt to the measurement environment by taking the skin stability for about 30 minutes in a space of constant temperature and humidity condition (temperature 20 ~ 24 ℃, humidity 40 ~ 60%) after changing the topcoat into the prepared gown. Then, after confirming the skin condition of the test subject, the back part of the test subject was wiped with 70% ethanol and dried. Then, four kinds of test product patches were attached and fixed to the test site (respectively). After the patch was removed 24 hours later, the subject was made to visit for a second time and the patch was removed. After 1 hour of patch removal, 2 experts confirmed skin irritation according to the criteria of International Contact Dermatitis Research Group (ICDRG) And the adverse reaction was confirmed. After the patch was removed 24 hours later, the third visit was made and skin irritation was confirmed according to the same criteria as the second visit and the adverse reaction was confirmed.

(2) Method of calculating skin irritation index

The ICDRG criteria are shown in the following table.

[Table 22]

The skin irritation index was calculated according to the following formula.

The skin irritation index calculated according to the above formula was classified according to the following table.

[Table 23]

(3) Evaluation of skin adverse reaction

To ensure the safety of the subjects, surveys and visual evaluations of adverse events were conducted at each visit, and the occurrence and symptom severity were checked. Adverse reactions and adverse events were determined in appropriate measures and participation of subjects in the test.

(4) Test results

All 32 subjects completed the test without dropping out.

SKIN, EMULSION, ESSENCE, Cream SKIN PEPTIDER Primary Stimulation Human body application test result was judged to be non-stimulant product as shown in the following table.

[Table 24]

In addition, no subjects reported skin adverse events (erythema, edema, stinging, itching, soreness, burning sensation, stiffness, tingling).

Claims (9)

&Lt; RTI ID = 0.0 &gt; decolorized &lt; / RTI &gt;
A cosmetic composition in the form of a skin, emulsion, essence or cream. The method according to claim 1,
Wherein said decolorization or deodorization is carried out using activated carbon. The method according to claim 1,
Wherein the cosmetic composition comprises 0.1 to 10% by weight of the decolorized or deodorized cord extract on the basis of the total weight of the cosmetic composition. The method according to claim 1,
Wherein the decolorized or deodorized Lard extract further comprises a high pressure dispersion treatment. 5. The method of claim 4,
Wherein the high-pressure dispersion treatment is carried out using a high-pressure homogenizer (microfluidizer). 5. The method of claim 4,
Wherein the high-pressure dispersion treatment is carried out using 1,3-butanediol. The method according to claim 1,
The cosmetic composition may be at least one selected from the group consisting of purified water, seawater, a moisturizing agent, a metal ion blocking agent, a pH adjusting agent, a surfactant, a skin softening agent, a moisture evaporation blocking agent, an antioxidant, a conditioning agent, a sterilizing preservative, Wherein the cosmetic composition further comprises one or more additives selected from the group consisting of an alcohol, a solubilizing agent, a flavoring agent, an oil, a synthetic high molecular substance, a neutralizing agent, a natural extract, a pigment, a pigment, a stabilizer, a vitamin and a chelating agent . The method according to claim 1,
The cosmetic composition may be selected from the group consisting of purified water, seawater, disodium edetate, maltose, glycerin, betaine, carbomer, tromethamine, glyceryl stearate, stearic acid, sorbitan sesquioleate, Citric acid, vegetable oil, hydrogenated vegetable oil, tocopheryl acetate, sunflower seed oil, fragrance, mash extract, succulent tea leaf extract, aloe vera leaf extract, acetyloctapeptide-3, 1,2-hexanediol, phenyl trimethicone, Panthenol, glycereth-26, sodium hyaluronate, ethanol, tamarind seed polysaccharide, butylene glycol, inulin, sorbitan monolaurate, Cetearyl alcohol, cyclopentasiloxane, dimethicone, meadowfoam seed oil, capryliliglycol, acetyloctapeptide-4, glycolipid and caprylic / capric triglyceride The cosmetic composition further comprises a binary of one or more additives selected from the group. The method according to claim 1,
Wherein the cosmetic composition has an effect of improving skin elasticity, improving dermal elasticity, improving elastic restoring force, improving skin pore, improving dermal density, or improving skin texture.

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