KR101906143B1 - A cosmetic composition for the extract of golden cocoon with improved anti-aging activity by far-infrared method - Google Patents

Abstract

The present invention provides a cosmetic composition for preventing skin aging comprising, as an active ingredient, a golden silkworm extract having enhanced skin wrinkle reducing effect, skin whitening effect, skin barrier improving effect and skin moisturizing effect by a far-infrared ray extraction method.

Description

[0001] The present invention relates to a cosmetic composition for preventing skin aging, which comprises an extract of golden cocoons having enhanced skin efficacy by a far-infrared ray extraction method,

The present invention relates to a method for increasing skin efficacy of a golden silkworm extract.

Conventionally, it has been generalized to contain extracts of natural materials in cosmetics related to skin efficacy, and various extracting methods and processing methods have been studied to obtain extracts having more excellent effects, including various effects, among such natural materials.

In the present invention, a far-infrared ray extraction method is used. Far infrared rays have long wavelengths (25 ~ 1,000㎛) of infrared rays. They are not visible to the naked eye and are well absorbed by materials. They are characterized by strong resonance and resonance of organic compound molecules. In addition, light generally has a good reflection when its wavelength is short, and it has a property of being absorbed when it reaches an object if its wavelength is long. Therefore, it has strong penetration and is used directly in human body for hospital treatment and esthetic. When applied to natural materials, the extracting method to be applied by utilizing the advantage of far-infrared ray light also activates the cell tissue by shaking the cell 2,000 times per minute when it comes in contact with the moisture and protein molecules constituting the cell, Extraction method by ordinary heat which is progressively heated by conduction phenomenon Far infrared ray extraction method that is extracted by heating by uniform heat due to self-heating inside the cell due to penetration of far-infrared rays deep inside, even if it is peeled from one direction. It is expected that the components can be extracted more efficiently.

Under these circumstances, the present inventors have used a variety of solvents and extraction methods from golden silkworm cocoons and have produced extracts from them, and have found that MMP-1 inhibition, collagen synthesis, melanin synthesis inhibition, cell activation effect, The present inventors have found that the present invention is superior to the extraction method used in the prior art.

* Prior art literature *

Patent Document 10-2014-0060749

The present invention is based on the finding that the far-infrared extracts of various extracting and processing methods are effective to inhibit MMP-1, collagen synthesis, inhibition of melanin synthesis, cell activity, It is confirmed that the moisturizing effect is superior to that of the conventional invention, thereby completing the present invention.

Accordingly, it is an object of the present invention to provide a cosmetic composition containing the golden silkworm extract extracted by a far-infrared ray extraction method as an active ingredient and having enhanced skin effect.

Another object of the present invention is to apply a cosmetic composition containing a golden silkworm extract extracted by the far-infrared ray extraction method to human skin.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention and claims.

The present invention provides a cosmetic composition for preventing aging of the skin, wherein the skin effect is enhanced by containing a golden silkworm extract extracted by a far-infrared ray extraction method as an active ingredient. The inventor of the present invention invented that the golden cocoon extract extracted by the far-infrared ray extraction method improves the efficacy about twice as much as the extract extracted by other extraction methods.

FIG. 1 is a graph showing cell survival rates at four concentrations for each example culled in order of efficacy according to the extraction method in Experimental Example 1. FIG.
FIG. 2 is a graph showing inhibition rates of MMP-1 expression at three concentrations for each of the examples culled in the order of potency by extraction method in Experimental Example 1. FIG.
FIG. 3 is a view showing a reduction rate of moisture loss by time in each example culled in order of efficacy according to the extraction method in Experimental Example 1. FIG.

The present invention provides a cosmetic composition for preventing aging of the skin, wherein the skin effect is enhanced by containing a golden silkworm extract extracted by a far-infrared ray extraction method as an active ingredient.

The silkworm silkworm, which is an active ingredient of the cosmetic composition of the present invention and has been transformed from a general silkworm cocoon and modified as a breed, has been reported to have superior skin effect than the conventional white silkworm cocoon -2014-0060749].

The golden silkworm cocoon used in the present invention can be obtained from larvae, pupa, silkworm, silkworm, silkworm, silk and the like, most preferably from silkworm cocoon. In addition, the golden silkworm extract used in the present invention can be obtained by various known extraction methods commonly used in the technical field of the present invention, preferably water, methanol, ethanol, propanol, butanol An anhydrous or a lower alcohol having 1 to 4 carbon atoms, a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, chloroform or 1,3-butylene glycol as an extraction solvent.

On the other hand, it is obvious that the extract of the present invention can obtain an extract showing substantially the same effect not only by using the above-mentioned extraction solvent but also by using other extraction solvent.

In addition, the gold cocoon extract used in the present invention includes not only the extract obtained by the above-mentioned extraction method, but also the extract which has undergone the conventional purification process after the far-infrared ray extraction. For example, by separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies (made for separation by size, charge, hydrophobicity or affinity), and the like, Fractions are also included in the extract of the present invention.

The extract of the present invention can be prepared in powder form by an additional process such as vacuum distillation and freeze-drying or spray-drying. In addition, the cosmetic composition of the present invention preferably contains 0.001 to 80.0% by weight of the golden cocoons extracted by the far-infrared ray extraction method with respect to the entire cosmetic composition. When the extract of the golden coco peat extract extracted by the far-infrared ray extraction method is used in an amount of less than 0.001% by weight based on the total weight of the composition, substantial skin improving effect of the cosmetic composition is hardly expected. When it is used in excess of 80.0% by weight, And it is not economical because there is little synergy effect with excess input.

The cosmetic composition containing the golden cocoon extract extracted by the far-infrared ray extraction method may be prepared into any of the commonly used formulations, and examples thereof include softening agents, convergent lotion, nutritional lotion, eye cream, A cream, a cleansing cream, a cleansing foam, a cleansing water, a powder, an essence, a pack, and the like.

Korean Patent Publication No. 10-2014-0060749 discloses an antioxidative effect extracted by an enzyme, an inhibitory effect on MMP-1 production, an inhibitory effect on MMP-1 production induced by ultraviolet ray, an effect of enhancing collagen synthesis, an effect of inhibiting melanin generation, Discloses a method for producing a cosmetic composition containing a golden silkworm extract having an effect of mitigating cytotoxicity, an effect of inhibiting the expression of inflammatory cytokines by ultraviolet irradiation, and an effect of improving skin elasticity.

Korean Patent Laid-Open Publication No. 10-2013-0108805 discloses a manufacturing method for a cosmetic composition for improving wrinkles, whitening, or moisturizing, which comprises an extract of a golden silkworm-derived substance extracted by hydrolysis as an active ingredient.

In another prior art, a golden silkworm, which is excellent in skin moisturizing, skin wrinkle alleviation, skin wrinkle alleviation, speckling alleviation, freckle alleviation, skin keratinization, alopecia reduction, hair loss mitigation, hangover prevention, Discloses a production method for a cosmetic composition containing an extract.

However, there is no report on the skin effect on the extract of the golden silkworm cocoon extracted using the far-infrared ray extraction method. Accordingly, the present invention has been accomplished by confirming that the extract of the golden silkworm extract promotes the skin effect in the far-infrared ray extraction method.

< Example >

Example  1-1 ~ 3: Preparation of golden cocoon extract (room temperature extraction)

Dried golden cocoons were crushed and 1000 g of dry weight was placed in a flask and each extraction solvent was applied as shown in the following Table 1 to immerse the raw material at room temperature so as to have a concentration of 10% (w / v), and the mixture was extracted for 1 day and filtered to obtain an extract .

Room temperature
extraction Heat number
extraction Ultrahigh pressure
extraction ultrasonic wave
extraction Far infrared ray
extraction After boiling, room temperature extraction Hydrolysis
(HCL) Purified water Example
1-1 Example
1-4 Example
1-7 Example
1-10 Example
1-13 Example
1-16 Example
1-19 50% ethanol Example
1-2 Example
1-5 Example
1-8 Example
1-11 Example
1-14 Example
1-17 - 100% ethanol Example
1-3 Example
1-6 Example
1-9 Example
1-12 Example
1-15 Example
1-18 -

Examples 1-4 to 6: Preparation of golden silkworm extract (hot water extraction)

The dried cocoons were crushed and the dried weight of 1000 g was placed in a flask. Each extract solvent was applied to the flask as shown in Table 1, so that the raw material was soaked in 10% (w / v) To obtain an extract.

Examples 1-7 to 9: Preparation of golden silkworm extract (ultra-high pressure extraction)

After drying the dried cocoons, 1000 g of dry weight is placed in a special pack for ultrahigh-pressure treatment, and each extraction solvent is applied as shown in Table 1, to thereby obtain a 10% (w / v) raw material. The dried powder of the golden silkworm cocoa was sealed in a special pack so that there was no air bubbles, and then it was put into an ultra high pressure reactor and subjected to ultra high pressure treatment at 300 MPa for 1 day to obtain an extract.

Examples 1-10 to 12: Preparation of golden silkworm extract (ultrasonic extraction)

Dried golden cocoons were pulverized and 1000 g of dry weight was placed in a flask and each extraction solvent was applied as shown in Table 1 to immerse the raw material in an amount of 10% (w / v) to prepare a sonicator (40 kHz, 40) Sonicator) for one day and filtered to obtain the extract.

Examples 1-13 to 15: Preparation of golden silkworm extract (far-infrared ray extraction)

Dried golden cocoons were crushed, and 1000 g of dried weight was placed in a flask, and each extraction solvent was applied as shown in Table 1 to immerse the raw material so as to have a concentration of 10% (w / v) And a far infrared ray having a wavelength of from 쨉 m to 1,000 쨉 m was added thereto, followed by extraction for one day and filtering to obtain an extract.

Examples 1-16 to 18: Preparation of golden silkworm extract (at room temperature after steaming)

Dried golden silkworm cocoons were crushed and dried, and the dried weight of 1000 g was immersed in a mesh net, and the mixture was shaken in a steamer at a temperature of 60 to 150 ° C for 10 minutes to 2 hours. The golden silkworm cooked powders thus prepared were immersed in 10% (w / v) of the raw material by applying each of the extraction solvents as shown in Table 1, and then extracted and filtered at room temperature for one day to obtain an extract.

Example 1-19: Preparation of golden silkworm extract (HCL)

The dried cocoons were pulverized, and 1000 g of hydrochloric acid was added thereto at a ratio of 1:10 (w / v), followed by hydrolysis at 100 for 3 hours to prepare a hydrolyzate. The hydrolyzate thus produced was filtered and neutralized by adding 3N sodium hydroxide thereto, followed by desalting to produce a silkworm hydrolyzate.

Experimental Example  1. Radical scavenging ability of golden cocoon extract DPPH Antioxidative Evaluation Using

The DPPH (2,2-diphenyl-1-picrylhydrazyl) radical in the antioxidant assay is dissolved in an organic solvent and has the maximum absorbance at 540 nm. The antioxidant dissolves the DPPH radical to change its color to yellow .

For the measurement of DPPH radical scavenging activity, the antioxidant activity of the golden silkworm extract extracted by the same procedures as in Examples 1 to 19 and the same amount of 0.01% of L-ascorbic acid as a positive control were measured for 30 minutes. The results of the free radical scavenging rate (%) were calculated by the following equation, and the results are shown in the following table.

Free radical scavenging ratio (%) = [1- (St-So) / (Bt-Bo)] x 100

St: absorbance at 540 nm after free radical scavenging of the sample solution

Bt: Absorbance at 540 nm after free radical scavenging of blank test solution

So: the absorbance at 540 nm before the reaction in the absence of the free radical of the sample solution

Bo: absorbance at 540 nm before the reaction in the absence of the free radical of the blank test solution

sample Radical scavenging ability (%) sample Radical scavenging ability (%) Example 1-1 83.3 Example 1-11 89.5 Examples 1-2 82.5 Examples 1-12 88.2 Example 1-3 80.9 Examples 1-13 99.2 Examples 1-4 95.4 Examples 1-14 98.7 Examples 1-5 95.2 Examples 1-15 97.6 Examples 1-6 94.1 Examples 1-16 87.8 Examples 1-7 95.1 Examples 1-17 86.2 Examples 1-8 94.8 Example 1-18 84.5 Examples 1-9 92.8 Example 1-19 83.6 Example 1-10 90.2 L-ascorbic acid 92.3

Experimental Example 2. Assessment of promoting keratinocyte differentiation of the golden silkworm extract

In this experimental example, the effect of the golden silkworm extract on cell proliferation was examined. 5.0, 10.0, 20.0 (%, w / v)) at a concentration of 5,000 cells / well in a 96 well plate incubator and cultured in a thermostat for 30 minutes. And cultured for 72 hours. Thiazoline blue was added and the cells were further cultured for 4 hours. After the culture medium was discarded, the reaction stop solution was added to each well of the microplate, and the mixture was stirred for 5 minutes, and the absorbance was measured at 570 nm. The control group was co-cultured with 10% Fetal Bovine Serum (FBS) medium as the optimal condition for cell growth, and the cell proliferation rate of the test sample was calculated with the cell proliferation of the control group as 100% . The measurement results are shown in Fig. 1 below.

As shown in FIG. 1, the effect of accelerating keratinocyte differentiation of gold cocoons used in the present invention was 154.02% at 20% concentration. In addition, the cell proliferation effect means that the golden silkworm extract does not have toxicity to the cells. From the cell proliferation effect test, the golden silkworm cocoon extract (1-13) of the present invention is superior to the extract It was confirmed that it is safe without cytotoxicity in addition to excellent cell proliferation effect.

Experimental Example 3. Evaluation of MMP-1 Inhibition Rate of Golden Silkworm Extract

Human dermal fibroblast neonatal (HDFn) was inoculated into a 96-well plate incubator at a concentration of 5 10 ⁴ cells / well, and then stabilized in a 37 incubator for 24 hours. Then, after rinsing twice with PBS, UVA was irradiated with 6 J / cm ² in the presence of PBS. Then, the culture was supplemented with 0.2% FBS supplemented with gold cocoon extract (25, 50, 100 / / ml) and 1 mM L-ascorbic acid as a positive control. After 48 hours, the supernatant was measured at 450 nm using a human total MMP-1 ELISA kit (human total MMP-1 Cat. No. DY901, R & D systems).

As a result, it was confirmed that all of the golden silkworm cocoons extract inhibited MMP-1 in a concentration-dependent manner. Among them, the golden silkworm extract (1-13) It was confirmed that MMP-1 inhibition rate is superior to that of L-ascorbic acid, which is a control group. [Figure 2]

Experimental Example  4. Evaluation of collagen synthesis increase rate of golden cocoon extract

Human normal fibroblasts were cultured in a 96 well plate incubator at a concentration of 1 × 10 ⁶ cells / well, and then stabilized under a medium supplemented with 10% FBS for 24 hours. Then, the golden cocoon extract was added to a final concentration of 100 μg / ml In serum-free DMEM medium for 48 hours. After the incubation, the supernatant of each plate was collected, and the amount of procollagen type I C-peptide (PICP) was measured in terms of ng / ml using a Collagen Kit (Takara, Japan) Were measured. As a comparative group, TGF-β (10 ng / ml, USA), which is known to have an effect of promoting collagen synthesis, was used as a comparative group. The rate of increase of collagen biosynthesis was calculated according to the following formula Are shown in Table 3 .

sample Increase in collagen biosynthesis (%) sample Increase in collagen biosynthesis (%) Example 1-1 84.2 Example 1-11 86.8 Examples 1-2 83.8 Examples 1-12 86.5 Example 1-3 81.0 Examples 1-13 99.9 Examples 1-4 98.9 Examples 1-14 99.3 Examples 1-5 98.5 Examples 1-15 99.1 Examples 1-6 97.8 Examples 1-16 86.1 Examples 1-7 88.3 Examples 1-17 85.5 Examples 1-8 88.1 Example 1-18 85.0 Examples 1-9 87.5 Example 1-19 97.0 Example 1-10 87.2 Positive control group
(TGF-β 10 ng / mL) 90.2

Table 3 shows the results of measuring the effect of enhancing collagen synthesis in the culture solution containing the golden silkworm extract and the positive control TGF-β by the extraction method using the fibroblast. The golden cocoon extract (1-13) of the present invention showed a higher rate of collagen biosynthesis when compared with the extracts extracted by other extraction methods at the same concentration of 0.01%.

Experimental Example  5. Skin moisturizing and barrier function recovery effect of golden cocoon extract

In order to examine the skin barrier function recovery effect of the cosmetic composition containing the golden silkworm extract of the present invention, the following experiment was conducted. Experiments were carried out by using a taping stripping method to damage the skin barrier and measuring the skin water loss (TEWL, transdermal water loss) to 4.0 g / m 2 / h measured by servomed (Sweden) evaporimeter EP 1 , And golden silkworm extract were applied to an area of 5 cm 2, respectively. After the skin moisture loss was measured after 2, 10, 20, and 60 minutes, the degree of decrease in barrier function was evaluated . At this time, all of the extracts extracted with ethanol were concentrated under reduced pressure and dissolved in water at the same concentration. As a positive control, hyaluronic acid sodium salt was used and water as an extraction solvent was used as a comparative group. The results are shown in FIG. 3 for comparison of changes over time, assuming skin moisture loss in the initial damaged barrier state as a reference (50%).

Experimental Example  6. Inhibitory Effect of Golden Cocoon Extract on Melanogenesis

In order to confirm the whitening effect of the golden cocoon extract of the present invention, experiments were carried out to examine the degree of inhibition of melanogenesis using B16F10 melanocyte-forming cells. B16F10 mouse-derived melanin-forming cells were cultured in a ^6- well plate incubator at a concentration of 2 × 10 ⁶ , and then treated with a golden silkworm extract (0.1%) at a concentration not causing toxicity and cultured for 72 hours. After incubation, the cells were recovered and the intracellular melanin was quantitatively modified by the Rotan method (Lotan: Cancer Res., 40: 3345-3350, 1980). The recovered cells were washed with PBS, homogenized solution was added, vortexed for 5 minutes, centrifuged, NaOH was added to the cell filtrate, absorbance was measured at 405 nm, melanin was quantified and the inhibition rate of melanin formation (%) was measured. Melanin formation inhibition rate (%) of melanin-forming cells of B16F1 was calculated by the following formula, and the results are shown in Table 4 below. Arbutin, a substance known to have an inhibitory effect on melanin production, was used as a comparative group.

Melanin formation inhibition rate (%) = [(A-B) / A] 100

A: Amount of melanin in wells to which no sample was added

B: Amount of melanin in the well to which the sample was added

sample Melanin inhibition rate (%) sample Melanin inhibition rate (%) Example 1-1 0.03 Example 1-11 0.08 Examples 1-2 0.01 Examples 1-12 0.05 Example 1-3 0.01 Examples 1-13 0.29 Examples 1-4 0.18 Examples 1-14 0.25 Examples 1-5 0.17 Examples 1-15 0.24 Examples 1-6 0.16 Examples 1-16 0.05 Examples 1-7 0.17 Examples 1-17 0.03 Examples 1-8 0.16 Example 1-18 0.03 Examples 1-9 0.15 Example 1-19 0.12 Example 1-10 0.10 Arbutin 0.03

As can be seen from the results of Table 4, it can be seen that the golden cocoon extract (1-13) of the present invention most inhibits melanin production in B16F10 melanin-forming cells. Extracts extracted by the far infrared ray extraction method showed better melanin formation inhibitory effect than arbutin, compared to arbutin, which is known to have a known whitening effect at the same concentration. Therefore, it can be seen that the cosmetic composition comprising the golden silkworm extract of the present invention has an excellent effect on inhibition of melanin formation.

Having described specific portions of the present invention in detail, those skilled in the art will appreciate that these specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (10)

A cosmetic composition comprising a golden silkworm extract extracted by a far-infrared ray extraction method as an active ingredient. 3. The method according to claim 1, wherein the golden silkworm extract
Crushing the dried cocoons of the golden cocoons and then putting a dry weight of 1000 g into the flask,
A step of applying an extraction solvent to the golden silkworm cocoa powder put into the flask and soaking the raw material so as to have a concentration of 10% (w / v)
Immersing the immersed gold cocoons powder in water at 70 to 100 ° C, adding a far infrared ray having a wavelength of 25 to 1,000 μm, extracting the powder for 1 to 10 days, and filtering to obtain an extract; . &Lt; / RTI &gt;
The cosmetic composition according to claim 2, wherein the extraction solvent of the cosmetic composition is selected from the group consisting of purified water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms, acetone, butylene glycol, dichloromethane, chloroform, ethyl acetate and benzene An extracting solvent or at least one extracting solvent.
The cosmetic composition according to claim 1, wherein the golden silkworm extract is used for preventing skin aging, and skin aging prevention is wrinkle improvement.
The cosmetic composition according to claim 1, wherein the golden silkworm extract is used for preventing skin aging, and skin aging prevention is skin whitening.
The cosmetic composition according to claim 1, wherein the golden silkworm extract is used for preventing skin aging, and the skin aging prevention is skin moisturizing effect.
The cosmetic composition according to claim 1, wherein the golden silkworm extract is used for preventing skin aging, and the skin aging prevention is a skin barrier improvement.
The cosmetic composition according to claim 1, wherein the cosmetic composition comprises an extract containing at least one of a caterpillar, a pupa, a cocoon, a silkworm, a silkworm, and silk.
The cosmetic composition according to claim 1, wherein the golden silkworm extract is contained in an amount of 0.001 to 80.0% by weight based on the total weight of the cosmetic composition.
The cosmetic composition according to claim 1, wherein the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, lotion, patch, mask sheet, soap, surfactant- , A wax foundation, and a spray.

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