KR100552269B1 - A cosmetic composition containing extract of Campsis grandiflora and/or Torreya nucifera seed - Google Patents

Abstract

The present invention relates to a cosmetic composition for external skin containing at least one selected from the genus hibiscus or non-extracted extract as a main active ingredient, and more specifically, a larva vine plant or a deciduous vine plant of Bignoniaceae. Anti-oxidant effect, skin aging effect, whitening effect and ultraviolet light, characterized in that it contains 0.001 ~ 30.0% by weight, preferably 0.01 ~ 5% by weight, as the active ingredient It relates to an external skin cosmetic composition excellent in alleviating the skin damage caused by the effect, the stimulating effect. According to the present invention, the extracts of Lilium vulgaris and Visage have not only antioxidant activity but also human fibroblast activity effect, skin fine wrinkle improvement effect, whitening effect, skin damage relieving effect and irritation relieving effect by using this substance. It can manufacture.

Gold leaf extract, Visa extract, Anti-aging cosmetic, Whitening effect, Skin irritation relieving effect, Skin external cosmetic

Description

A cosmetic composition containing extract of Campsis grandiflora and / or Torreya nucifera seed}

The present invention relates to a cosmetic composition for external skin containing at least one selected from the genus hibiscus or non-extracted extract as a main active ingredient, and more specifically, a larva vine plant or a deciduous vine plant of Bignoniaceae. Anti-oxidant effect, skin aging effect, whitening effect and ultraviolet light, characterized in that it contains 0.001 ~ 30.0% by weight, preferably 0.01 ~ 5% by weight, as the active ingredient It relates to an external skin cosmetic composition excellent in alleviating the skin damage caused by the effect, the stimulating effect. According to the present invention, the extracts of Lilium vulgaris and Visage have not only antioxidant activity but also human fibroblast activity effect, skin fine wrinkle improvement effect, whitening effect, skin damage relieving effect and irritation relieving effect by using this substance. It can manufacture.

Human skin undergoes various physical and chemical changes during the aging process. The causes are largely divided into internal aging and photo-aging, and research on this has been actively conducted. Free radicals may be activated by UV rays, stress, disease conditions, environmental factors, wounds, and age. If this condition is exacerbated, it destroys antioxidant defenses in vivo and damages cells and tissues. And aging. More specifically, lipids, proteins, polysaccharides, and nucleic acids, which are major constituents of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging. In particular, the oxidation of proteins is caused by severe cuts in the connective tissues of collagen, hyaluronic acid, elastin, proteoglycan, fibronectin, and so on, resulting in severe over-inflammatory reaction and skin elasticity. If this worsens, mutations in the DNA can lead to mutations, cancer, and reduced immune function.

Therefore, it protects the cell membrane by eliminating the free radicals caused by metabolic processes of the body, irradiation by UV irradiation and inflammatory reactions, and the already damaged cells should be regenerated by active metabolism to proliferate the cells. You can recover quickly and maintain healthy skin. Aging involves not only free radicals, but also an enzyme called matrix metalloprotease (MMP) .In vivo, the synthesis and degradation of extracellular substrates such as collagen are properly regulated, but their synthesis decreases as aging progresses. Expression of matrix metalloproteinase (MMP), which is an enzyme that degrades, is promoted, thereby decreasing the elasticity of the skin and forming wrinkles. Ultraviolet irradiation also activates these degrading enzymes. Therefore, there is a need for the development of a substance capable of regulating MMP expression that inhibits activation or inhibiting its activity in cells. Until now, most of the raw materials used as materials for cosmetics simply inhibited enzyme activity. Therefore, we tried to regulate the expression and activity of MMP expression induced by intracellular natural aging and photoaging.

Next, the skin changes due to pigmentation. Factors that determine skin color are basically the areas such as tanning due to blemishes, freckles and sun exposure, or overall pigmentation, other than acne, scars, keratin distribution, Blood circulation, stress, and health. Among the above factors, the most important factor is pigmentation. The pigments that affect skin color include melanin, melazoids, carotene, and hemoglobin, the most important of which is melanin. The biggest factor affecting the biosynthesis of melanin is ultraviolet rays and hormones in the body. Melanin plays an important role in absorbing or scattering ultraviolet rays to prevent damage to the skin from ultraviolet rays. There is no special maximum absorption wavelength and it absorbs light in all areas. In addition, it is excellent in removing the active oxygen species, but sometimes melanin itself generates free radicals, and other substances are reduced or oxidized by catechol or quinone in the melanin structure, and melanin itself shows the properties of free radicals. Pray.

The biosynthesis of melanin is a series of oxidation processes, starting with the conversion of tyrosine, an amino acid, from the melanocytes of melanocytes to tyrocinase and dihydroxy phenylalanine. eumelanin). This biosynthesis process is carried out in a special type of brown intracellular organelles called melanosomes. Melanosomes, including melanin granules, move from the periphery of the nucleus to the dendritic end of the dendritic projections and into the cytoplasm by the phagocytosis of keratinocytes. Accumulate around the nucleus.

The synthesis of melanin, the number of melanosomes, and migration to the surrounding keratinocytes are partially influenced by hormones and ultraviolet rays, and primarily by genetic effects. In addition, it is known that metal ions such as cytokines, copper, zinc, and iron, interferon, prostaglandin, histamine, etc., which are involved in the expression of tyrosinase, the synthesis and transfer of melanin, are involved.

Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A 6-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-93150), and some extracts are already tyrosinase. Although it is used as a whitening cosmetic because it has inhibitory activity, its use is restricted due to its low stability in cosmetic formulations, which is degraded or colored, causing off-flavor, efficacy at the biological level, unclear effect, and safety problems. . And the inhibitory effect of tyrosinase has been demonstrated, but the effect is low in experiments similar to the actual biologic level. Therefore, the evaluation of the inhibitory effect by melanoma cell culture similar to the biological level is required. In addition, hydroquinone is defined as a carcinogenic substance and its use is prohibited or restricted in cosmetics. Kojic acid and ascorbic acid are very unstable substances. Cosmetics containing a small amount of the above components occur browning at room temperature for several weeks.

In order to solve the problem of the skin, a lot of cosmetics using natural products have recently been developed to reduce skin irritation caused by various chemicals. Natural materials have less side effects on the skin, and as the consumer's response to cosmetics using natural materials has increased recently, the development value of cosmetics as a raw material for cosmetics is increasing. Therefore, the present inventors have studied the possibility of applying to cosmetics in a variety of natural products that have not been known so far, select the deciduous vine plants of Bignoniaceae and the evergreen tree of the evergreen tree of the Taxaceae Extracts were prepared from them, and the results of measuring skin antioxidant effects, collagen synthesis effects, skin fine wrinkles relieving effect, whitening effect, UV damage relieving effect and skin irritation relieving effect were excellent. Was done.

Geumdeunghwa is a deciduous vine plant of the dicotyledon plant Tung-Tung-Hwa, the scientific name Campsis grandiflora , also known as Jacaranda. It is native to China and has adsorbed roots on its branches. The leaves face each other and are odd one-time quills. 7 ~ 9 small leaves, egg-shaped or egg-shaped basso, 3 ~ 6cm long, with sharp tip, with serrated hairs on the edge. Flowers bloom in August-September and form 5 cones in conical inflorescences at the end of branches. The diameter of the flower is 6 ~ 8cm, the color is orange, and the inside is orange. Calyxes are 3cm long, divided into 5 pieces, and the fragments are lancet shaped and pointed. Corollas are bell-shaped, similar to funnels. Fruits are capsules, square, split into two and ripen in October. It has been planted in sub-central districts and also for ornamental purposes. In herbal medicine, it was used for fecal bulge, skin ulcer, acne, amenorrhea, mild fever, abdominal pain, blood fever.

The jasmine tree is an evergreen tree of the herbaceous genus yew, and its scientific name is Torreya nucifera . Similar species include Cephalotaxus koreana Nakai and Cephalotaxus koreana var. Nana Nak.

Distributed in Korea and Japan, it grows in deep and fertile sandy loam. An evergreen coniferous tree of male oyster, egg-shaped bark is grayish brown, its stem spreads in all directions, and the tree's tree is shallowly separated. The leaves are usually 25mm long and 3mm wide, feather-shaped, leathery, linear, sharp, hairless, dark green on the back, brown on the back, but green on the middle and edges, only on the back. There are yellowish white pore lines on both sides of the posterior vein of the back. The leaf disease is 3mm long and falls in 6-7 years. Seeds are nucleus (核果) and ripen red in October of the following year. The seeds are 2 ~ 3cm long and 1 ~ 2cm in diameter. Peeled and dried dried outer seed skin of the seed is called Torreyae Semen . Since ancient times, wood has been used as a building material, furniture material, sculptures, and it can be used as memorial water, park water, bonsai, and only seeds are called visas. It is also used for food (snacks, gangjeong) and oil.

The present invention is to determine the efficacy of the deciduous vines of the deciduous vine plant of the Jacaranda family or the extract of the jasminoides of the evergreen tree of evergreen as a raw material of skin cosmetics, and to develop a method of using the same By using this raw material, it aims at providing the skin external cosmetics which have predetermined | prescribed functionality.

That is, an object of the present invention is to provide a natural extract that is applicable to skin cosmetics and exhibits excellent anti-aging effects such as antioxidant effects, collagen synthesis effects, skin fine wrinkles effects when contained in cosmetics.

It is also an object of the present invention to provide a natural extract exhibiting a whitening effect by ultraviolet irradiation, a skin damage alleviation effect by ultraviolet rays, and a skin stimulation effect by an inflammatory mediator when applied to skin cosmetics.

In order to achieve the above object, the present inventors confirmed the skin improvement effect of the gold lantern, non-extract extract and used to prepare the external skin cosmetics using them.

The present invention relates to a cosmetic composition for external skin containing at least one selected from a hibiscus extract or non-visa extract as a main active ingredient.

In addition, the present invention is characterized in that the gold hydrangea extract or non-visa extract contains 0.001 to 30.0% by weight based on the whole composition. When the content of the extract is less than 0.001% by weight does not appear to improve the skin, when the content of more than 30.0% by weight is not economical because the degree of effect increase for the content is insignificant.

In addition, the present invention is characterized by using at least one selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane or hexane as the extraction solvent.

In another aspect, the present invention provides a cosmetic composition for external skin, characterized in that the extract is obtained by cold washing at room temperature, heat filtration, and further obtained by concentrating the solvent under reduced pressure or lyophilization.

In addition, the present invention, the skin cosmetic composition is characterized in that it has an antioxidant, anti-aging, anti-wrinkle, whitening, skin damage suppression and irritation relief function by ultraviolet light.

In addition, the present invention, the cosmetic composition for external skin is selected from cosmetic composition or ointment of the lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) type or water-in-oil (W / O) type formulation. It features.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only illustrative description of the present invention and the scope of the present invention is not limited to these Examples.

Example 1 Preparation of Geumdeunghwa Extract

After distillation, the dry gold lanterns were refluxed three times for 5 hours with 95% (V / V) ethanol aqueous solution, and then cooled and then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C or lower. The extract was dissolved / dissolved using one or more solvents selected from purified water, ethanol, butylene glycol, and propylene glycol so as to contain 0.001-30.0% by weight of the reduced pressure concentrate and the lyophilisate.

Example 2: Preparation of Visa Extract

The dry visa was refluxed three times for 5 hours with an aqueous 95% (V / V) ethanol solution, and the mixture was cooled and then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C or lower. Dispersed / dissolved non-extracts were prepared using one or more solvents selected from purified water, ethanol, butylene glycol, and propylene glycol so as to contain 0.001-30.0% by weight of the vacuum concentrate and the lyophilisate.

Example 3: Antioxidant Effect Measurement Experiment Using NBT Method

In this Example, to compare the antioxidant effect of the hibiscus extract and non-extract extract obtained in Examples 1, 2 and other antioxidants, such as retinol and BHT (Butylated Hydroxytoluene) under laboratory conditions, the sample was compared The antioxidant activity was measured using the method.

In order to measure the antioxidant effect, the active oxygen produced by xanthine and xanthine oxidase is measured by the NBT method, and the effect of the test substance removing the active oxygen, that is, the active oxygen scavenging effect, is evaluated. Free radicals are produced by xanthine and xanthine oxidase. This active oxygen reacts with Nitro Blue Tetrazolium (NBT) to measure the active oxygen scavenging rate by measuring the blue color produced by this at a wavelength of 560 nm.

As a measuring method

0.05M Na ₂ CO ₃ ------------------------------------ 2.4ml

 2.3mM xanthine solution ---------------------------------- 0.1ml

 3.3mM EDTA Solution ------------------------------------ 0.1ml

 4.BSA solution ---------------------------------------- 0.1ml

 5.0.72 mM NBT solution ---------------------------------- 0.1ml

 6.Xanthine Oxidase Solution ----------------------------- 0.1ml

7.6 mM CuCl ₂ solution ----------------------------------- 0.1ml

① Add 1,2,3,4,5 to Bayer's bottle and add 0.1ml of sample solution to it and leave at 25 ℃ for 10 minutes.

② Add 6 and stir quickly, and start incubation for 20 minutes at 25 ℃.

③ After that, add 7 to stop the reaction and measure the absorbance St at 560nm.

④ For the blank test, measure the absorbance Bt by using distilled water instead of the sample solution as above.

⑤ Also, the blank of the sample solution is operated in the same manner using distilled water instead of 6 to measure the absorbance Bo.

The result of the effect was calculated by Equation 1, the results are shown in Table 1.

% Inhibition = [1- (St-So) / (Bt-Bo)] X 100

St: absorbance at 560 nm after enzyme reaction of sample solution

Bt: absorbance at 560 nm after enzymatic reaction of blank test solution

So: absorbance at 560 nm before reaction without enzyme addition of sample solution

Bo: absorbance at 560 nm before reaction without enzyme in blank test solution

As a result, as shown in Table 1, it was confirmed that the excellent antioxidant effect in all the samples tested at 0.1% concentration. It was confirmed that the extract of Goldfinaceae has excellent antioxidant power similar to retinol and BHT, and the non-extract extract showed better antioxidant effect than retinol and BHT at 0.1% concentration.

 Sample Name  Treatment concentration (%)  Antioxidative effect(%)  Goldback Extract (Example 1)  0.1  92  Visa extract (Example 2)  0.1  98  Retinol  0.1  93  BHT  0.1  90

Example 4 Experiment for Measuring Antioxidant Effect Using DPPH Method

This Example is to compare the antioxidant activity such as green tea extract and vitamin E in laboratory conditions to determine the antioxidant effect of the gold and yellow extract obtained in Examples 1 and 2 by using the DPPH method Measured.

The DPPH method measures the antioxidant activity by reducing power using a free group called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The free radical scavenging rate is measured at a wavelength of 560 nm by comparing the degree to which the DPPH is reduced by the test substance to reduce the absorbance.

As a reagent used, 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) 0.1 mM solution was dissolved in methanol to make 100 ml.

As a measuring method

① Add 0.15ml of sample solution to 0.15ml of 0.1mM DPPH solution in 96-well plate, stir rapidly, and incubate for 10 minutes at 25 ℃.

(2) Then measure the absorbance St at 560nm.

③ In the blank test, measure the absorbance Bt by using distilled water instead of the sample solution as above.

④ Again, the blank of the sample solution was operated in the same manner using methanol instead of the 0.1 mM DPPH solution to measure the absorbance Bo.

The result of the effect was calculated by Equation 2, the results are shown in Table 2.

% Inhibition = [1- (St-So) / (Bt-Bo)] X 100

St: absorbance at 560 nm after free radical scavenging of sample solution

Bt: absorbance at 560 nm after free radical scavenging of blank test solution

So: Absorbance at 560 nm before reaction without free radicals in sample solution

Bo: absorbance at 560 nm before reaction without free radicals in blank test solution

As a result, as shown in Table 2, the extract of Visa was found to have excellent antioxidant power similar to retinol and BHT at 0.01% concentration, and the gold leaf extract had a much better antioxidant effect than the retinol and BHT at 0.01% concentration.

 Sample Name  Treatment Concentration (%)  Antioxidative effect(%)  Goldback Extract (Example 1)  0.01  82  Visa extract (Example 2)  0.01  30  Retinol  0.01  20  BHT  0.01  23

Example 5 Measurement of MMP-1 Enzyme Inhibitory Effect

Substrate metalloproteinase (MMP-1) inhibitory activity was determined in a biochemical model based on the use of collagen and gelatin conjugated to purified collagenase and its substrate, fluorescein (EnzChek ™ gela) Tinase / collagenase kit, molculaprobe). Collagenase purified from Clostridium historiescom was supplied in the EnzChek ™ gelatinase / collagenase kit. A reaction buffer consisting of DQ-collagen purified from porcine skin and conjugated to fluorescein with 0.05M Tris-HCl, 0.15M NaCl, 5mM CaCl ₂ and 0.2mM sodium azide (pH 7.6) was EnzChek (TM) gelatinase. Collagenase kit (molecular probes) was used. Goldfinch and Visage extracts were dissolved in the reaction buffer. It is 4; 2; 0.4; 0.2; Test at 0.1% (w / v). Dilutions of the test extracts were incubated with 25 ug / ml of DQ-collagen and 0.1 U / ml of collagenase for 15 minutes, 45 minutes, and 120 minutes at room temperature.

In each experimental condition, the control corresponding to the collagenase and DQ-collagen mixture was also incubated in the same manner. Blanks, hereinafter referred to as 'enzyme-free blanks', were also incubated in the presence of DQ-collagen and in the absence of collagenase at each experimental condition. Each experiment was performed three times.

After 15, 45, and 120 minutes, the signal corresponding to the degradation of DQ-collagen was measured with a fluorometer (excitation: 485 nm, emission: 505 nm). The fluorescence value of each sample was measured based on the 'enzyme free blank' fluorescence value. Results are expressed as percent variance for fluorescence units per sample and control.

In conclusion, under the selected experimental conditions, gold and green extracts tested at 0.04 to 4% (V / V) retained anticollagenase / gelatinase activity in a dose dependent manner.

The results are shown in Table 3. The non-extract extract inhibited 80% of the collagen degrading activity of Clostridium collagenase when treated with 0.04%, which was superior to the inhibitory effect of green tea extract, and the gold leaf extract also exhibited 45% of the collagen degrading activity of 0.04%. By inhibiting the green tea extract showed a similar degree of inhibitory activity.

 Experimental Sample  Treatment concentration (%, V / V)  % Inhibition  Goldback Extract (Example 1) 0.02% 26 0.04% 45  Visa extract (Example 2) 0.02% 55 0.04% 80  Retinol 0.02% 0 0.04% 5  Green Tea Extract 0.02% 52 0.04% 56

Example 6: Evaluation of MMP-1 Expression Inhibition by UV Irradiation

This Example was subjected to ELISA to measure the concentration of MMP-1 after UV irradiation and sample addition of the gold and non-visa extract obtained in Examples 1 to 2.

Human dermal fibroblasts are irradiated with UVA at an energy of 5 J / cm 2 using a UV chamber. UV irradiation amount and incubation time were established through preliminary experiments to maximize the expression of MMP in fibroblasts. Negative controls were wrapped in tinfoil and kept in the UVA environment for the same time. UVA emission was measured using a UV radiometer. The cells during the UVA irradiation are intact with the previously dispensed medium, and are irradiated with UVA and then exchanged with the medium containing the sample for 24 hours of incubation, and then the medium is recovered and coated in 96-well. The primary antibody (MMP-1 (Ab-5) monoclonal antibody and MMP-2 (Ab-3) monoclonal antibody) is treated and reacted at 37 ° C. for 60 minutes. After reacting the secondary antibody, an anti-mouse IgG (whole mouse, alkaline phosphatase conjugated) for about 60 minutes, the alkaline phosphatase substrate solution (1 mg / ml -nitrophenyl phosphate in diethanolamine buffer solution) was reacted at room temperature for 30 minutes and then the microplate reader was reacted. Measure the absorbance at 405 nm. As a control, the sample without addition is used.

Inhibition of MMP-1 expression induced by UV irradiation showed more than 20% inhibition of the hibiscus extract and non-extract extract compared to the control group, which is superior to the inhibition rate of retinol used as a control.

 Test group  Treatment concentration (%)  MMP-1 expression inhibition rate (%)  Control  -  -  Goldback Extract (Example 1)  0.1  25  Visa extract (Example 2)  0.1  21  Retinol  0.1  18

Example 7: Determination of melanin production inhibitory effect using B16F1 melanogenesis cells

This Example is to determine the whitening effect by looking at the degree of inhibition of melanin production on B16F1 melanocytes in order to confirm the whitening effect of the extracts and non-extracted extract obtained in Examples 1 and 2.

B16F1 melanocytes used in this example are cell strains derived from mice, and cells that secrete a black pigment called melanin. Samples were treated during the artificial culture of these cells to evaluate the degree of melanin black pigment reduction. B16F1 melanocytes used in this example were used by receiving from the ATCC (American Type Culture Collection, Accession No .: 6323).

The melanin biosynthesis inhibitory effect of B16F1 melanogenesis cells was measured as follows. B16F1 melanocytes were dispensed in 6-well plates at a concentration of 2 X 10 ⁶ per well, and the cells were attached and incubated for 72 hours by treating the sample at a concentration that does not cause toxicity. After 72 hours of incubation, the cells were detached with trypsin-EDTA, the cell number was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was performed by slightly modifying the method of Rotan: Cancer Res. , 40: 3345-3350, 1980. The cell pellet was washed once with PBS, and then 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt cells. Melt the extracted melanin by adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes), and measure the absorbance of melanin at 405 nm with a microplate reader. Production inhibition rate (%) was measured. Melanin inhibition rate (%) of B16F1 melanogenesis cells was calculated by the equation (3), IC ₅₀ value is the concentration of the substance inhibiting 50% melanogenesis.

% Inhibition = [(A-B) / A] X100

A: Melanin amount in wells without sample

B: Melanin amount in the wells to which the sample was added

The IC ₅₀ value of the test extract was 0.08%, and the IC ₅₀ value of the non-extract extract was 0.1%, indicating that the B16F1 melanocyte-forming melanin-producing effect was 0.1%. It showed a similar or superior effect compared to the extracts of the lettuce extract (Table 5).

 Sample Name  Treatment concentration (%) Inhibitory effect of melanin synthesis (IC ₅₀ )  Goldback Extract (Example 1)  0.1  0.08%  Visa extract (Example 2)  0.1  0.1%  Hydroquinone  0.1  0.03%  Arbutin  0.1  0.2%  Lettuce extract  0.1  5%  Soluble Licorice Extract  0.1  0.03%

Example 8: Cytotoxic Alleviation Effect by UV Irradiation

This example was carried out to evaluate the mitigating effect of cytotoxicity by ultraviolet irradiation of the hibiscus extract and non-extract extract obtained in Examples 1 and 2. Fibroblasts (fibroblasts) were placed in a 24-well test plate ⁵ times 1 × 10 ⁵ and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with 10 mJ / cm 2 of UV light using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), after which PBS was removed and 10% FBS was added to the cell culture medium (DMEM). 1 ml) was added. Herein, the treatment was performed for 24 hours after treatment of the extract and Visage extract to be evaluated. After 24 hours, the medium was removed, and 500 μl of culture medium and 60 μl of MTT solution (2.5 mg / ml) were added to each well, followed by incubation in a 37 ° C. CO ₂ incubator for 2 hours. The medium was removed, and 500 μl of isopropanol-HCl (0.04N) was added thereto. After shaking for 5 minutes, the cells were lysed and 100 μl of the supernatant was transferred to a 96-well test plate, followed by measurement of 565 nm absorbance in a microplate reader. Cell survival rate (%) was measured by the equation (4) and cytotoxicity relaxation by ultraviolet light was calculated by the equation (5).

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

Bo: 565 nm absorbance of wells that only developed cell culture media

Bt: 565 nm absorbance of the wells that developed the colorless reaction wells

St: 565 nm absorbance of the wells that reacted the sample treated wells

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

Bo: cell viability of wells without UV irradiation and no sample treatment

Bt: cell viability of wells that were not irradiated with UV light

St: Cell survival rate of wells treated with UV

Experimental results showed that the extract of Goldfingerum inhibits the cytotoxicity caused by UV rays by 26% at 0.013% concentration, and the non-extract extract mitigates the cytotoxicity of 59% at 0.013% concentration, effectively defending the cytotoxicity caused by UV rays. . Through the above experiments, it can be seen that cell damage caused by ultraviolet rays is effectively protected at low concentration (Table 6).

 Sample Name  Treatment concentration (%)  Cytotoxic alleviation rate (%)  Goldback Extract (Example 1) 0.013 26 0.025 27  Visa extract (Example 2) 0.13 59 0.025 72

Example 9 Inhibitory Effect of Inflammatory Cytokine Expression by UV Irradiation

In this example, 24-year-old fibroblasts isolated from human epidermal tissues were evaluated in order to evaluate the inhibitory effect of inflammatory cytokine expression expressed by UV irradiation of the extracts of gold and green extracts obtained in Examples 1 and 2. Four ⁵ × 10 ⁴ cells were placed in a well test plate and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with 10 mJ / cm 2 of ultraviolet rays using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), after which PBS was removed and cell culture medium (FMEM was not added to DMEM). Medium) 350 μl was added. After treatment with the gold lantern extract and the non-extract extract to be evaluated therein was incubated for 5 hours. 150 µl of the culture supernatant was taken to quantify IL-1α, and the inhibitory effect of inflammatory cytokine expression was determined by the extracts of Lilium vulgaris and non-leaved extracts. The amount of IL-1α was quantified using Enzyme-linked Immunosorbent Assay, and the production rate of IL-1α was calculated by Equation 6.

Inhibitory rate of inflammatory cytokine expression (%) = [1- (St-Bo) / (Bt-Bo)] X 100

Bo: IL-1α production in wells without UV irradiation

Bt: IL-1α production in wells that were not irradiated with UV light

St: IL-1α production amount of wells that were irradiated with UV light

Experimental results showed that the extract of gold larvae inhibits the production of inflammatory cytokines, IL-1α, by 58% at a concentration of 0.013%, effectively preventing the occurrence of inflammation caused by UV at low concentrations (Table 7).

 Sample Name  Treatment concentration (%)  Inhibitory rate of inflammatory cytokine expression (%)  Goldback Extract (Example 1) 0.013 58 0.025 80  Visa extract (Example 2) 0.13 5 0.025 7

Examples 10, 11 and Comparative Example 1

The present Example was prepared by the cosmetics containing the extracts and non-extractable extracts obtained in Examples 1 and 2 to evaluate the skin elasticity improvement effect in a comparative experiment with Comparative Example 1.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 8. First, the phase b) recorded in Table 8 is heated and stored at 70 ° C. To this, a) phase is added, and after pre-emulsification, it is emulsified uniformly with a homomixer, and then cooled slowly to prepare a cream (Examples 10, 11 and Comparative Example 1). For 20 experimenters (20-35 year old female), the creams prepared in Examples 10 and 11 were applied to the right side of the face and the creams prepared in Comparative Example 1 were applied twice a day for 3 consecutive months to the left side of the face. It was.

The skin elasticity improvement effect after the completion of the experiment was measured using a skin elasticity measuring instrument (cutometer SEM 575, C + K Electronic Co., Germany) before and after using the product for 2 months. The experimental results are described in Table 9 below as the ΔR8 value of the cutometer SEM 575, where the R8 value represents the nature of the viscoelasticity of the skin. As shown in Table 8, it can be seen that the skin elasticity improving effect of the tester coated with the cream containing the hibiscus extract and non-extract extract.

 Raw material  Example 10  Example 11  Comparative Example 1  end  Stearyl alcohol  8  8  8  Stearic acid  2  2  2  Stearic cholesterol  2  2  2  Squalane  4  4  4  2-octyldodecyl alcohol  6  6  6  Polyoxyethylene (25 mol addition) alcohol ester  3  3  3  Glyceryl monostearic acid ester  2  2  2  I  Goldback Extract (Example 1)  One  - -  Visa extract (Example 2) -  One -  Propylene glycol  5  5  5  Quantity  Quantity  Quantity  Quantity

Note) Unit: Weight%

 Experimental product  Skin elasticity effect (△ R8)  Example 8  0.21  Example 9  0.26  Comparative Example 1  0.12

n = 20, p <0.05

Examples 12-13 and Comparative Example 2

The present Example was prepared by the cosmetics containing the extract and non-extracted extract obtained in Examples 1 and 2 to evaluate the skin whitening effect in humans in Comparative Example 2dhk comparative experiments.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 10. First, the phase b) recorded in Table 10 is heated and stored at 70 ° C. To this, a) phase is added, and after pre-emulsification, it is emulsified uniformly with a homomixer, and then cooled slowly to prepare a cream (Examples 12, 13 and Comparative Example 2). 20 experimenters (women aged 20-35 years) were treated with creams prepared in Examples 12 and 13 on the right side of the face, and creams prepared in Comparative Example 2 on the left side of the face for 2 consecutive days, 2 times a day. Applied. After completion of the test, the skin of the applied areas on both sides of the face was measured using an image analyzer and the color of the skin using chromameter (Minolta CR300) to measure the change in brightness of color (△ L). Subjective visual observation was conducted to determine the effect according to the following classification. The results are shown in Table 11 below. At this time, the degree of whitening efficacy was classified into the following seven ratings.

Whitening efficacy evaluation criteria:

-3: very worse, -2: worse, -1: slightly worse,

0: no change, 1: slightly improved, 2: improved, 3: greatly improved

As shown in Table 11, it can be seen that the whitening effect is excellent in the facial skin of the experimenter coated with the cream containing the hibiscus extract and non-visa extract.

 Raw material  Example 12  Example 13  Comparative Example 12  end  Stearyl alcohol  8  8  8  Stearic acid  2  2  2  Stearic Acid Cholesterol  2  2  2  Squalane  4  4  4  2-octyldodecyl alcohol  6  6  6  Polyoxyethylene (25 mol addition) alcohol ester  3  3  3  Glyceryl monostearic acid ester  2  2  2  I  Goldback Extract (Example 1)  One  - -  Visa extract (Example 2) -  One -  Propylene glycol  5  5  5  Quantity  Quantity  Quantity  Quantity

Note) Unit: Weight%

 Skin color change (L)  Objective evaluation of the skilled person  Subjective Evaluation of the Subject  Example 12  Example 13  Comparative Example 2  Example 12  Example 13  Comparative Example 2  Example 12  Example 13  Comparative Example 2  medium  5.32  4.82  2.03  2.9  2.3  1.9  2.7  2.2  1.7

Other examples are shown below. That is, the lotion, milky lotion and essence containing the golden yellowing extract and the non-extract extract obtained in Examples 1 and 2 were prepared in Examples 14 to 19. The lotion, milky lotion and essence containing these extracts and non-extracted extracts had excellent effects on skin improvement such as antioxidant effect, collagen synthesis promoting effect, skin fine wrinkle improvement effect and skin damage inhibition or irritation relief function by UV light. These hibiscus extracts and non-extract extracts can be added to formulations of lotion, gel, water soluble liquid, cream, essence, oil-in-water (O / W) and water-in-oil (W / O) forms to prepare cosmetics. It can be easily applied in the technical field to which it belongs, but the content of the present invention is not limited thereto.

Example 14 Preparation of Lotion Containing the Goldback Extract Obtained in Example 1

To 8 g of 95% ethanol, 0.05 g of polypyrrolidone, 0.1 g of oleyl alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of fragrance, 0.1 g of paraoxybenzoic acid methyl ester, a small amount of antioxidant, and a small amount of pigment are dissolved. . 0.05 g of goldfinch extract obtained in Example 1 and 5 g of glycerin were dissolved in 85.33 g of purified water, and then the mixed solution was added and stirred to obtain a lotion having a skin improvement effect.

Example 15 Preparation of Lotion Containing the Visage Extract Obtained in Example 2

To 8 g of 95% ethanol, 0.05 g of polypyrrolidone, 0.1 g of oleyl alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of fragrance, 0.1 g of paraoxybenzoic acid methyl ester, a small amount of antioxidant, and a small amount of pigment are dissolved. . 0.05 g of the non-extract extract obtained in Example 2 and 5 g of glycerine were dissolved in 85.33 g of purified water, and then the mixed solution was added and stirred to obtain a lotion having a skin improvement effect.

Example 16 Preparation of an Milky Milk Containing the Goldfinch Extract Obtained in Example 1

1.2 g of cetyl alcohol, 10 g of squalane, 2 g of petrolatum, 0.2 g of paraoxybenzoic acid ethyl ester, 1 g of glycerin monoesteralide, 1 g of polyoxyethylene (20 mole added) monooleate and 0.1 g of fragrance were mixed and dissolved at 70 ° C. , 0.5 g of distilled gold extract obtained in Example 1, 5 g of dipropylene glycol, 2 g of polyethylene glycol-1500, 0.2 g of triethanolamine, and 76.2 g of purified water were dissolved by heating to 75 ° C. Both mixtures were emulsified and cooled to obtain an emulsion having an oil-in-water (O / W) type skin improvement effect.

Example 17 Preparation of Latex Containing the Visage Extract Obtained in Example 2

1.2 g of cetyl alcohol, 10 g of squalane, 2 g of petrolatum, 0.2 g of paraoxybenzoic acid ethyl ester, 1 g of glycerin monoesteralide, 1 g of polyoxyethylene (20 mole added) monooleate and 0.1 g of fragrance were mixed and dissolved at 70 ° C. , 0.5 g of the non-extract extract obtained in Example 2, 5 g of dipropylene glycol, 2 g of polyethylene glycol-1500, 0.2 g of triethanolamine, and 76.2 g of purified water were heated to 75 ° C. to dissolve it. Both mixtures were emulsified and cooled to obtain an emulsion having an oil-in-water (O / W) type skin improvement effect.

Example 18 Preparation of Cosmetic Liquid Containing the Goldfinch Extract Obtained in Example 1

5 g of 95% ethyl alcohol, 1.2 g of polyoxyethylene sorbitan monooleate, 0.3 g of kitulose, 0.2 g of sodium hyaluronate, 0.2 g of vitamin E-acetate, 0.2 g of sodium licorice, 0.1 g of paraoxybenzoic acid ethyl ester, 1 g of the sternum extract obtained in Example 1 and a suitable amount of pigment were mixed to obtain a cosmetic solution having an effect of improving skin.

Example 19 Preparation of Cosmetic Liquid Containing the Visage Extract Obtained in Example 2

5 g of 95% ethyl alcohol, 1.2 g of polyoxyethylene sorbitan monooleate, 0.3 g of kitulose, 0.2 g of sodium hyaluronate, 0.2 g of vitamin E-acetate, 0.2 g of sodium licorice, 0.1 g of paraoxybenzoic acid ethyl ester, 1 g of the non-extract extract obtained in Example 2 and an appropriate amount of pigments were mixed to obtain a cosmetic liquid having a skin improvement effect.

As described above, the extract according to the present invention has an excellent anti-aging effect such as antioxidant effect, MMP inhibitory effect, MMP expression control effect by UV irradiation, fine wrinkle improvement effect, and relieve skin irritation caused by UV irradiation. Indicated.

In addition, the hibiscus and Visa extract showed a whitening effect such as melanin production inhibitory effect, which is a substance causing blemishes, freckles and skin pigmentation.

Therefore, cosmetic compositions such as lotion, cream, milky lotion, packs, powders, etc. containing such extracts and non-extracted extracts have excellent skin externalizing effects such as antioxidant effect, collagen degrading activity control effect, skin fine wrinkle improvement effect, anti-inflammatory and whitening effect. It can be seen that.

Claims (6)

Cosmetic composition for external use of skin containing at least one selected from the hibiscus extract or non-extract extract as the main active ingredient. The method of claim 1, wherein the jinhwahwa extract or Visage extract contains 0.001 to 30.0% by weight based on the whole composition, the skin external containing at least one selected from the hwahwahwa extract or Visage extract as the main active ingredient Cosmetic composition. The gold lamp according to claim 1 or 2, wherein at least one selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane or hexane is used as an extraction solvent. Cosmetic composition for external skin containing at least one selected from the extract or non-visa extract as the main active ingredient. The method according to claim 1 or 2, wherein the extract is obtained by cold condensation at room temperature, heating and filtration, and further obtained by concentrating the solvent under reduced pressure or lyophilization. Cosmetic composition for external skin containing the above as the main active ingredient. According to claim 1, wherein the external cosmetic composition for skin has the function of anti-aging, anti-wrinkle, whitening, skin damage by irritation and stimulation alleviation, at least one selected from the gold leaf extract or visa extract Cosmetic composition for external skin, containing as an active ingredient. The cosmetic composition according to claim 1 or 2, wherein the cosmetic composition for external skin is a cosmetic composition of a lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) type or water-in-oil (W / O) type or A cosmetic composition for external application of skin, comprising one or more selected from gold hydrangea extract or non-visa extract as a main active ingredient, which is selected from ointments.