KR20190010299A - Cosmetic composition for comprising Siegesbeckia pubescens - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin moisturizing or improvement of skin inflammation, and more specifically, to a composition which is excellent in skin soothing, skin moisturizing or skin inflammation improvement effect. The composition comprises Siegesbeckia pubescens extract or diterpene compounds having kauran skeleton isolated from the Siegesbeckia pubescens extract as an active ingredient.

Description

[0001] Cosmetic composition for making Siegesbeckia pubescens [0002] The present invention relates to a cosmetic composition for improving skin soothing, skin moisturizing or skin inflammation,

The present invention relates to a cosmetic composition for improving skin soothing, skin moisturizing or skin irritation, and more particularly, to a cosmetic composition for skin soothing, skin moisturizing, or skin irritation which contains, as an active ingredient, a diterpene compound isolated from an extract To a cosmetic composition for improving inflammation.

When exposed to the external environment, the skin plays a very important role in protecting the body from physical and chemical stimuli and pathogens. The skin section is divided into three layers: epidermis, dermis, and subcutaneous tissue. Among them, the stratum corneum of the epidermis contains about 10 to 20% of water and is present at the outermost part of the human body to suppress water evaporation from the body, Lt; / RTI > However, due to various stresses from environmental changes, lifestyle, and social environment, environmental pollution caused by pollutants, frequent cleansing due to makeup habits, and natural skin degeneration caused by aging, the skin becomes dry, the surface becomes rough, The problem of skin health becomes more and more important.

As a method of maintaining skin health from skin stress, many efforts have been made to develop a moisturizing material for preventing drying of skin moisture and improving water retention ability in stratum corneum. When the dry skin is formed, the barrier function of the skin surface is lost, and it is not only easy to penetrate the skin of the irritant or allergen from the outside, but also cause the skin to reject the penetration.

Skin rejection is caused by activation of keratinocytes, keratinocytes, which constitute the majority of the surface components, by a small number of cells such as Langerhans and melanocytes, and cytokines produced and released by these cells (cytokine). The cytokine acts to attract inflammatory cells. In particular, it stimulates constitutive cells of the skin to cause an inflammatory reaction mainly in the upper layer of the skin and to form an eczema reaction, which is called non-allergic skin disease.

Therefore, there is a possibility that a substance having a strong skin-soothing and skin-moisturizing effect can be extended to an anti-inflammatory effect and conversely, a substance having an anti-inflammatory effect can be expected to be effective for skin soothing and skin moisturizing, There has been a problem that has to go through a very difficult process.

On the other hand, the present inventors have separated various diterpene derivatives from "comedium", which is a prelude to a hairy origin or a contest of a composite plant (Chemical Constituents of Sigesbeckia pubescens Herb, Thesis, Master's Degree, Seoul National University, Feb. 2009). It is an annual plant growing at a height of 40-100 centimeters. It is common in mountains and fields. It has dense short hairs all over. It has a pointed end and sawtoothed leaves on the stem. It grows in August-September, but at the end of the stem, a yellow flower runs in a flower bed, and the fur on the outer surface of the five spatula-shaped scabbins surrounding the fruit sticks to other objects with sticky liquid. It has been used for the treatment of limb paralysis, hypertension, rheumatism and the like because it has the effect of eliminating (removing) customs from the herb, strengthening the muscles and bones, and lowering the blood pressure.

However, it has been known that the extract is effective for improving skin damage, promoting wrinkles, promoting hair growth, etc., and has not been studied to be effective for skin soothing, skin moisturizing or skin irritation improvement.

The present inventors have searched various compounds derived from natural materials for substances having skin soothing, skin moisturizing or skin irritating effects. As a result, it was found that the extract of the comedium contains a specific diterpene derivative having a kauran skeleton among various diterpene derivatives, and these substances have a skin soothing effect, a skin moisturizing effect or a skin inflammation improving effect.

Accordingly, it is an object of the present invention to provide a cosmetic composition for skin soothing, skin moisturizing or skin irritation improvement comprising a comedium extract containing a diterpene derivative as an active ingredient.

The present invention provides a cosmetic composition for skin soothing, skin moisturizing or skin irritation improvement comprising a comed extract as an active ingredient.

Also, the present invention provides a cosmetic composition for skin soothing, skin moisturizing or skin inflammation improvement wherein the comed extract comprises at least one diterpene compound represented by the following general formulas (1) to (7).

    &Lt; Formula 1 > < EMI ID =

        &Lt; Formula 3 > < Formula 4 >

     &Lt; Formula 5 > < EMI ID =

In the present invention, it is preferable that the flavor extract is extracted with water, C ₁ -C ₆ alcohol or a mixture thereof, and the diterpene compound is preferably contained in an amount of 0.0001 to 20% by weight based on the total weight of the composition .

The cosmetic composition may be a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, The composition may be formulated into various forms such as a soap, a shampoo, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion, a body cleanser, a latex, a press powder, a loose powder, ), An animal placenta-derived protein, betulinic acid and chlorella extract, as well as skin wrinkle-improving ingredients.

In another embodiment, the present invention provides a method for preparing a cosmetic composition for skin soothing, skin moisturizing or skin inflammation improvement, which comprises the step of adding the extract and / or the diterpene compound.

According to the present invention, a specific diterpene derivative having a kauran skeleton isolated from the extract of the present invention and the extract, that is, a compound represented by the general formulas (1) to (7) is used as a cosmetic composition to provide skin soothing, skin moisturizing or skin inflammation There is an effect to improve.

Fig. 1 shows the moisture content with time according to the concentration of the flavor extract.
Fig. 2 shows the amount of collagen production in human skin fibroblasts by concentration of the comedium extract.
Fig. 3 shows the melanin inhibitory activity by concentration of the comedium extract.
Figure 4 shows the anti-allergic activity according to the concentration of the extract.
Fig. 5 shows cytotoxicity according to the concentration of the tryptic extract.

Hereinafter, preferred embodiments of the present invention will be described in detail. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the invention as claimed. It will be obvious to those who have knowledge of. In the following description of the present invention, detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

It has been found by the present inventor that the diterpene compounds of the kaurans family isolated from the comed extract and the extracts, namely the compounds of the formulas (1) to (7), are effective for improving skin soothing, skin moisturizing or skin inflammation.

    &Lt; Formula 1 > < EMI ID =

        &Lt; Formula 3 > < Formula 4 >

     &Lt; Formula 5 > < EMI ID =

It is preferable that the flavor extract is extracted with water, a C ₁ -C ₆ alcohol, or a mixed solvent thereof, more preferably a mixture obtained by extraction with a mixed solvent of water and methanol. As the mixed solvent of water and methanol, for example, 70 to 90% by volume, preferably about 80% by volume of methanol may be used. The extraction method may be carried out by a conventional method, for example, extraction with stirring at room temperature for 2 to 5 hours, or extraction using a high frequency extractor for 2 to 5 hours. If necessary, the extraction may be carried out at a temperature (for example, about 40 캜 to 60 캜), and the extraction may be performed once or plural times (for example, about 3 times). The obtained extract can be dried by a conventional drying method, for example, vacuum drying, hot air drying and the like.

The compounds of the formulas (1) to (7) are known substances and can be isolated from the compound according to a known method. In other words, the method reported by the present inventors (Chemical Constituents of Sigesbeckia pubescens Herb, Master Thesis, Seoul National University, Feb. 2009) or Jiang X, Yunbao M and Yunlong X 1992) Diterpenoids from Siegesbeckia pubescens. Phytochemistry 3 917-921, or a method disclosed in Published Public Publication No. 10-1026070 and the like.

The diterpene compound is preferably contained in an amount of 0.0001 to 20% by weight based on the total weight of the composition.

The composition of the present invention can be formulated in the form of a cosmetic composition. The cosmetic composition includes the above-mentioned comic extract or a comedium extract containing the compounds of the above formulas (1) to (7) as an active ingredient, and its form is not limited. That is, the form of the cosmetic composition of the present invention may be in the form of a usual cosmetic composition such as cream, pack, lotion, essence, lotion, foundation and make-up base. , &Lt; / RTI &gt; For example, the skin lotion, the skin softener, the skin toner, the astringent, the lotion, the milk lotion, the moisturizing lotion, the nutrition lotion, the massage cream, the nutrition cream, the moisturizing cream, And may be formulated into formulations such as mask sheets, exfoliants, soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, emulsions, press powders, loose powders and eye shadows.

 The content of the above-mentioned trynaerian extract or the comedium extract containing the compounds of the above-mentioned formulas 1 to 7 contained in the unit cosmetic composition may vary depending on the form of extract / compound and the like, for example, 100 nM to 1 mM per unit cosmetic May range from 1 to 1000 [mu] M, preferably from 1 to 200 [mu] M.

In addition, it may further include skin wrinkle improving components such as vitamin C, retinoic acid, transforming growth factor (TGF), animal placenta-derived proteins, betulinic acid and chlorella extract.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example  1> Celebration  Extract preparation

5 kg of the dried mixture was added to 10 L of 70% methanol (water: ethanol = 30: 70, v / v) and extracted with a high frequency extractor at room temperature for 3 hours. The extract was filtered to obtain a filtrate, and the remaining residue was further extracted twice with 70% ethanol added thereto in the same manner as above. The filtrates were combined and dried under reduced pressure at 40 ° C to obtain 433 g of a tryptic extract.

< Example  2> Skin moisturizing  Efficacy evaluation

(1) Measurement of skin capacitance

The skin moisture content of the stratum corneum was measured by measuring the capacitance of the stratum corneum to evaluate the skin moisturizing effect of the extract of the present invention. The healthy subjects were selected and the test group containing 2%, 3%, and 4% (v / v) of the test extracts were added to the test strips at 4 hours and 6 hours after the test. The skin's electrical capacity was measured using a moisture checker MY-808S (Scalar, Tokyo) and the results were expressed as moisture content (%).

As shown in FIG. 1 and Table 1 below, the water content after 4 hours and 6 hours in the experimental group supplemented with 3% of the tryna extract was 20% and 18%, respectively, about 12% higher than those of the other experimental groups and the control group And it was confirmed that it exhibited excellent moisturizing effect.

After 4 hours After 6 hours Control 17 17 2% 16 15 Rice extract 3% 20 18 Flavor extract 4% 17 16

(2) Moisture measurement using moisture meter

Moisturizing power was measured using a moisture meter (Corneometer CM825, Courage + Khazaka, Germany) in order to evaluate the moisturizing effect of the extract of the present invention. As a comparative group, Sasa extract, Jujube extract, Third-order distilled water was used. Ten women in their twenties were divided into 2 × 2 ㎝ ² and 8 compartments per each sample. After washing several times with water, the water was removed without stimulation. After 15 minutes, 10 μl of each sample was applied evenly to each of the two compartments. After 2 hours, measurements were made five times per compartment using a moisture analyzer. The experiments were carried out in a constant temperature room with room temperature of 21 ± 2 ℃ and relative humidity of 50 ± 5%. The mean values of the five measurements were used, and the mean values of the five subjects were shown in Table 2 below. The calculation formula of the moisturizing power and the moisture increasing rate is as follows.

Corneometer value = S - ΔS

(S: corneometer value before sample application, ΔS: corneometer value after 2 hours of sample application)

Moisture power increase rate (%) = Experimental group Moisture power - Control group Moisture power

As a result of the experiment, the moisture content of the extract of the present invention was the highest at 42%, and the moisture increase rate of 40% was found to have an excellent moisturizing effect.

Moisture power (%) Growth rate (%) Sashou extract 32.4 8 Jujube extract 35.3 18 Sulfur urging extract 33.4 12 Extract 42 40 Control group (distilled water) 29.8 -

< Example  3> Collagen production experiment

Human dermal fibroblasts (HDFn) were inoculated in a culture medium at a concentration of 5 × 10 ⁴ cells / well in a 48-well plate and cultured at 37 ° C and 5% CO ₂ . After 24 hours, the culture broth was cultured for 24 hours in a fresh medium containing 10%, 50, 100 μg / ml of colchicine extract. The cultures were collected and analyzed by using an enzymes-linked immunoassay kit of Takara Co. The amount was measured. A standard curve between collagen concentration of 0 / / ㎖ and 640 / / ㎖ was obtained using 640 ㎍ / ㎖ of collagen solution, and the amount of collagen in the medium was calculated. The results are shown in Fig. 2 and Table 3 below.

As a result of the experiment, it was confirmed that the extract of the present invention has the effect of improving the wrinkle by the effect of increasing the collagen production by 23% at the concentration of 100 ㎍ / ml of the extract of the present invention.

control 10 [mu] g / ml 50 [mu] g / ml 100 [mu] g / ml C-terminal peptide
(PIP,% of control) 100 103 114 123 Viability (% of control) 99 99 99 99

Further, in order to evaluate the wrinkle-reducing effect on the diterpene compound of the cauren series extracted from the comedium, a collagen production experiment was carried out in the same manner as in the above comedium extract. At this time, the concentration of diterpene compound (the above Chemical Formulas 1 to 7) was 100 ppm, and the experimental results are shown in Table 4 below. As a result, the diterpene compound showed an effect of increasing collagen production by about 39%, confirming that the diterpene compound according to the present invention is excellent in improving wrinkles.

control Formula 1 (2) (3) Formula 4 Formula 5 6 Formula 7 C-terminal peptide
(PIP,% of control) 100 143 138 140 142 135 137 141 Viability (% of control) 99 99 99 99 99 99 99 99

< Example  4> Evaluation of whitening efficacy

B16-F1 murine melanoma cells were seeded at a concentration of 1 × 10 ⁵ cells / well in a 6-well plate using DMEM (Dulbecco's Modified Eagle's Medium) medium containing 10% FBS (fetal bovine serum) Under% CO ₂ conditions, the cells were cultured until approximately 80% of the cells were attached to the well bottom. After the culture, the medium was removed and the tryptic extract was diluted with the medium (10, 50, 100 ㎍ / ㎖) and diluted. The cells were cultured at 37 ° C and 5% CO ₂ . After the culture, the medium was removed and the cells were washed with PBS (phosphate buffered saline) and treated with trypsin to recover the cells. The recovered cells were counted using a hematocytometer, centrifuged at 5,000-10,000 rpm for 10 minutes, and the supernatant was removed to obtain a pellet. The cell pellet was dried in a 60 ° C. dry oven, and 100 μl of 1 M sodium hydroxide solution containing 10% by weight of DMSO or cell lysis buffer was added thereto to obtain intracellular melanin in a 60 ° C. thermostat. The absorbance of the melanin solution was measured at 405 nm using a microplate measuring device to determine the amount of melanin per cell constant or the amount of melanin per constant protein. The results are shown in FIG. 3 and Table 5 below.

Melanin formation was reduced by 20% when the concentration of the extract was 100 μg / ml, as shown in FIG. 3, and thus the extract of the present invention was found to contain It was confirmed that the whitening effect was excellent.

control 10 [mu] g / ml 50 [mu] g / ml 100 [mu] g / ml Intra-Melanin (% of control) 100 99 90 80 Viability (% of control) 100 101 102 100

In addition, the whitening efficacy evaluation of the diterpene compound of the kauran series extracted from the comedium was carried out in the same manner as the above-mentioned comedium extract. At this time, the concentration of diterpene compound (the above Chemical Formulas 1 to 7) was 100 ppm, and the experimental results are shown in Table 6 below. As a result, melanin formation in the diterpene compound was inhibited by about 28%, confirming that the whitening effect of the diterpene compound according to the present invention was excellent.

control Formula 1 (2) (3) Formula 4 Formula 5 6 Formula 7 Intra-Melanin (% of control) 100 73 71 74 72 71 71 72 Viability (% of control) 100 100 100 101 100 102 100 101

< Example  5> Anti-allergy  Efficacy evaluation

RBL-2H3 cells were suspended in DMEM containing 10% by weight of FBS and inoculated at a concentration of 2 × 10 ⁵ cells / well in a 24-well plate. The cells were sensitized with mouse monoclonal IgE 100 ㎍ / ㎖ in each well and cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours. Cells were washed with 500 μl of tyrode's buffer, treated with 10, 50, and 100 μg / ml of concentrated extract, and incubated for 15 min at 37 ° C in a 5% CO ₂ incubator. DNP-BSA 100 Mu] g / ml. The supernatant was treated with 50 μl of 1 mM p-nitrophenyl-N-acetyl- substrate, incubated at 37 ° C in a 5% CO ₂ incubator for 1 hour, and absorbance was measured at 405 nm using an ELISA reader. As shown in FIG. 4 and FIG. 7, 21% anti-allergic effect was observed in the group treated with 100 μg / ml of the extract of the present invention, compared to the control without the extract of the present invention.

control 10 [mu] g / ml 50 [mu] g / ml 100 [mu] g / ml β-hexosaminidase release (% of control) 100 99 90 80

< Example  6> Anti-inflammatory cytotoxicity assay

For the anti - inflammatory cytotoxicity assay, human dermal fibroblast (HDF) was used for MTT assay and the absorbance was measured at 570 ㎚ ~ 690 ㎚. First, fibroblasts were inoculated into each well of a 96-well plate at a concentration of 1 × 10 ⁴ cells / well and cultured in DMEM medium at 37 ° C. for 24 hours. Then, the control group was changed to DMEM medium containing 2.5% of serum with 0.5%, 1%, 2% and 4% concentration of the extract, and the control group was replaced with serum-free DMEM medium for 24 hours Respectively. After cultivation, MTT solution (3 mg / ml) was added to measure cell viability, and the cytotoxicity was calculated by the following formula.

Cytotoxicity (%) = (absorbance at the time of treating the extract / control absorbance) × 100 (%)

As shown in FIG. 5 and Table 8 below, cytotoxicity tests on human skin cells according to concentrations (0.5%, 1%, 2% and 4%) of the comedium extract showed no cytotoxicity up to 2% .

Extract concentration cell 0.5% E 1.0% E 2% E 4% E Cytotoxicity (%) 0 -70 -48 89 68

< Example  7> Skin soothing  Efficacy evaluation

In order to evaluate the skin soothing effect of the diterpene compounds of the kaurans family extracted from the comedones, the inhibitory effect of the inflammatory cytokine (IL-1α) on HaCaT cell (human keratinocyte cell), which is a human skin keratinocyte, was analyzed.

HaCaT cells were inoculated in DMEM medium supplemented with 10% FBS at a concentration of 2 × 10 ⁵ cells / well in a 24-well plate, and cultured in a 5% CO ₂ incubator at 37 ° C. for 24 hours. After the medium was removed from the plate, the cells were washed once with PBS (phosphate buffered saline), exchanged with serum-free DMEM medium to which no FBS was added, and then the diterpenes of the above formulas (1) to (7) ppm for 24 hours, treated with sodium dodecyl sulfate (SDS), and cultured for another 4 hours. After 4 hours, a portion of the culture was taken and used for IL-1α quantification and protein quantification. The quantification of IL-1α was performed according to the kit's IL-1α assay using the human IL-1α kit from Endogen Co. The amount of protein was quantitated by the BCA method of Sigma, and the quantified IL- The amount of IL-1? Secretion per cell was measured. Ketoprofen, which has been used as a skin irritation mitigating agent, was used as a positive control, and the final IL-1α production rate (%) was calculated by the following equation. This is the percentage of IL-1α produced in each sample when the amount of IL-1α was 100 when SDS alone was administered to the skin cells. The results are shown in Table 9 below.

IL-1? Production rate (%) = (IL-1? Production amount of sample / IL-1? Production amount of sample treated only with SDS)

 This test was performed to analyze the biosynthesis of IL-1α which can cause erythema on the skin through SDS treatment. The IL-1α production amount in the diterpene compounds of the above Chemical Formulas 1 to 7 was determined by measuring the amount of IL- 1α production. As a result, it can be seen that the effect of relieving troubles such as erythema occurring on the skin is great, which means that the skin soothing effect is excellent.

IL-1? Production rate (%) Control group 34.42 ± 1.78 Sample treated only with SDS 100.00 Formula 1 68.11 ± 3.63 (2) 67.05 + - 4.80 (3) 68.83 + - 3.65 Formula 4 70.01 + - 4.92 Formula 5 69.21 + - 5.03 6 67.17 + - 3.05 Formula 7 69.64 ± 2.89 Ketoprofen 64.12 + - 4.66

< Example  8> Manufacturing cosmetics

(1) Manufacture of skin lotion

0.1% by weight of a tryptic extract, 1.0% by weight of beta-1,3-glucan, 2.0% by weight of butylene glycol, 2.0% by weight of propylene glycol, 0.1% by weight of carboxyvinyl polymer, 0.2% Skin lotion was prepared by mixing 0.4 wt% of Bait 80, 10.0 wt% of ethanol, 0.1 wt% of triethanolamine, 0.05 wt% of preservative, 0.1 wt% of fragrance and a balance of purified water.

(2) Nutrition cream manufacturing

0.2% by weight of the flavor extract, 5.0% by weight of beta-1,3-glucan, 3.0% by weight of butylene glycol, 3.0% by weight of propylene glycol, 0.2% by weight of triethanolamine, 10.0% 2.0% by weight of hardened castor oil, 0.5% by weight of sorbitan sesquioleate, 10.0% by weight of liquid paraffin, 5.0% by weight squalane, 5.0% by weight of caprylic / capric triglyceride, 5.0% by weight of glycerin, 0.05% , 0.1% by weight of perfume, and purified water were mixed to prepare a nutritional cream according to a conventional method.

The foregoing description is merely illustrative of the present invention, and various modifications may be made without departing from the essential characteristics of the present invention. Accordingly, the embodiments disclosed herein are intended to be illustrative rather than limiting, and the spirit and scope of the present invention is not limited by these embodiments. The scope of protection of the present invention should be construed according to the claims, and all the techniques within the scope of the same should be construed as being included in the scope of the present invention.

Claims (9)

A cosmetic composition for skin soothing, skin moisturizing or skin irritation improvement comprising an extract of Comvitae as an active ingredient
The cosmetic composition according to claim 1, wherein the flavor extract comprises at least one diterpene compound represented by the following general formulas (1) to (7)
&Lt; Formula 1 >< EMI ID =

&Lt; Formula 3 &gt;&lt; Formula 4 &gt;

&Lt; Formula 5 &gt;&lt; EMI ID =

The cosmetic composition according to claim 1, wherein the flavor extract is extracted with water, C ₁ -C ₆ alcohol, or a mixed solvent thereof
The cosmetic composition according to claim 2, wherein the diterpene compound is contained in an amount of 0.0001 to 20% by weight based on the total weight of the composition
The composition according to claim 2, wherein the composition is at least one selected from the group consisting of a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, And is characterized in that it is made of any one of the formulations selected from the group consisting of mask sheet, exfoliating agent, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder and eye shadow Cosmetic composition
The composition according to claim 2, wherein the composition further comprises at least one skin wrinkle-improving ingredient selected from the group consisting of vitamin C, retinoic acid, transforming growth factor (TGF), animal placenta-derived protein, betulinic acid and chlorella extract A cosmetic composition
A method for preparing a cosmetic composition for skin soothing, skin moisturizing or skin irritation, which comprises the step of adding, as an active ingredient, a comedium extract comprising at least one of diterpene compounds represented by the following formulas
&Lt; Formula 1 >< EMI ID =

&Lt; Formula 3 &gt;&lt; Formula 4 &gt;

&Lt; Formula 5 &gt;&lt; EMI ID =

8. The method of claim 7,
Wherein the diterpene compound is contained in an amount of 0.0001 to 20% by weight based on the total weight of the composition [8] The method of claim 7, wherein after the adding step, at least one skin wrinkle-reducing component selected from the group consisting of vitamin C, retinoic acid, transforming growth factor (TGF), animal placenta-derived protein, betulinic acid, and chlorella extract Comprising the steps of: &lt; RTI ID = 0.0 &gt;

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