KR20140039370A - Cosmetic composition including hydrolysate from silkworm gland - Google Patents

Abstract

The present invention relates to a cosmetic composition including a hydrolysate from a silkworm silk gland. A cosmetic composition of the present invention is characterized in including a hydrolysate from a silkworm silk gland, and more specifically, the hydrolysate is prepared by separating a gland from a silkworm, dissolving the separated gland into water, centrifuging and obtaining a supernatant, adding an alkaline compound to conduct hydrolysis, neutralizing and desalting. According to the present invention, provided is a cosmetic composition comprising a hydrolysate from a silkworm silk gland which is harmless to humans and having good antioxidant activity, dermal fibroblast proliferating properties, skin affinity, etc.

Description

[0001] The present invention relates to a cosmetic composition comprising a hydrolyzate derived from silkworm gum,

The present invention relates to a cosmetic composition comprising a hydrolyzate derived from a silkworm gum, and more particularly, to a cosmetic composition comprising a silkworm hydrolyzate derived from a silkworm gum which is harmless to the human body and is excellent in antioxidant ability, skin cell proliferating ability and skin affinity.

Conventionally, cosmetic compositions used in cosmetic packs are mostly composed of artificial synthetic compositions. Therefore, the effect of damaging fragile skin or regenerating the skin with healthy and rich skin is inferior.

In severe cases, it has serious disadvantages that can cause side effects such as skin rash and allergies. Accordingly, in recent years, attempts have been made to use natural products having a moisturizing effect and a whitening effect on cosmetic products. However, the natural products known to date can be expected to have some degree of skin whitening effect, but they do not show satisfactory characteristics in terms of moisture absorption and moisturizing power.

The proteins that make cocoon are fibroin and sericin. Among them, sericin has excellent cell proliferation effect and can be used as a serum substitute. It is known to be a main ingredient of cosmetic products because it has effects of skin coating, moisturizing, antioxidant ability and skin affinity.

However, sericin obtained by a conventional method known to date can not be used as a raw material for a cosmetic as a raw material as a collagen material, and conventional sericin separation and recovery methods can be carried out by using a harmful solvent such as lithium bromide (LiBr) , There is a problem in that the process of obtaining such a complicated and low yield is performed by using high temperature heating of 90 占 폚 or more.

As a related prior art, Korea Patent No. 10-0494229 (registered on May 31, 2005, name: Sericin manufacturing method and mask pack using the same) and Korea Patent No. 10-0542266 (registered on Jan. 2006 2006 March 03, name: cosmetic pack composition containing sericin protein extracted from silkworm cocoon).

It is an object of the present invention to provide a method of extracting silkworm glands from silkworm glands, which is isolated from silkworm glands and can be obtained at a high yield through a simple process of extracting at room temperature without using a harmful solvent and is excellent in antioxidant ability, skin cell- And to provide a cosmetic composition comprising the hydrolyzate.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

To achieve the above object, the cosmetic composition of the present invention is characterized by comprising a hydrolyzate derived from silkworm.

The hydrolyzate is characterized in that the hydrolyzate is obtained by separating the false gland from the silkworm, dissolving the separated gland in water, centrifuging the supernatant, adding the alkali to the supernatant, hydrolyzing and then neutralizing and desalting.

The hydrolyzate is characterized in that sericin and fibroin are hydrolyzed.

The hydrolyzate is contained in an amount of 0.001 to 50% by weight based on the weight of the cosmetic composition.

The hydrolysis is performed at 15 to 30 占 폚.

It is characterized in that NaOH is used as the alkali.

Phosphoric acid is used in the neutralization process.

The cosmetic composition is characterized by having formulations of lotion (skin lotion), nutrition lotion, nutritional cream, massage cream, or nutritional essence.

The present invention provides a cosmetic composition comprising a hydrolyzate derived from a silkworm-derived silkworm which is capable of obtaining a hydrolyzate by extraction at room temperature without using a harmful solvent.

Also provided is a cosmetic composition comprising a silkworm-derived hydrolyzate derived from silkworm germ, which is excellent in antioxidant ability, skin cell growth potential, skin affinity, and the like.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the antioxidant ability of the hydrolyzate of the present invention according to the difference in solvent. FIG.
FIG. 2 is a graph showing the cytotoxicity test results according to the concentration of the hydrolyzate added. FIG.
3 is a graph showing the effect of hydrolyzate on human skin cell proliferation in serum-free medium.
FIG. 4A is a diagram showing cell attachment and proliferation according to hydrolyzate coating concentration in a cell culture dish. FIG.
FIG. 4B is a graph showing cell attachment and proliferation effects according to the hydrolyzate coating concentration in a cell culture dish. FIG.
5 is a view showing a result of skin irritation test of the hydrolyzate.
A: rabbit head left "Health (insomnia) skin - test substance (hydrolyzate of Example 1)"
B: rabbit head right "scratch and skin-test substance (hydrolyzate of Example 1)"
C: Rabbit head left "Scratch and skin - no treatment"
D: rabbit hair right "Health (non-tanning) skin - no treatment"

The terms, techniques, and the like described in this specification are used in the meaning commonly used in the technical field to which the present invention belongs, unless otherwise specified.

Hereinafter, the cosmetic composition comprising the hydrolyzate derived from the silkworm mucilage of the present invention will be described in detail as follows.

1. Preparation of hydrolyzate

A hydrolyzate obtained from silkworm gland is prepared.

The hydrolyzate is obtained by hydrolyzing a silk protein in which the fibroin and sericin are mixed and the silk protein is dissolved at room temperature. In particular, the hydrolyzate is water-soluble to be easily dissolved in water to be.

Thus, the hydrolyzate means a mixture of substances obtained by hydrolysis of sericin and fibroin, and includes peptides and amino acids.

The hydrolyzate may be any as long as it is separated from silkworm gland secretion. Preferably, the hydrolyzate is one isolated from silkworm ultrafine powder.

The method of obtaining the hydrolyzate will be described in detail as follows.

First, seals are separated from silkworms. At this time, the silkworm is preferably lyophilized to separate the false glands.

Species of the silkworm include Bombix mori Although all species considered to be silkworms capable of obtaining false glands in general can be used, they are excellent in cell proliferation effect and can be obtained by reducing the breeding time and labor cost to obtain hydrolysates It is preferred that the silkworm be used for the silkworm.

Second, the supernatant obtained by dissolving the separated glands in water and centrifuging them is used.

At this time, it is preferable that the false glands are processed into ultrafine powder. Preferably, the lyophilized silk gland is milled and then processed into an ultrafine powder.

This is because when the false gland is in an ultrafine powder state, dissolution can be easily performed at room temperature (15 to 30 ° C) so as to minimize component changes.

The reason for solubilizing the false gland in water in the state of ultrafine powder is that it dissolves well by maximizing the contact area with water molecules at the time of dissolution, and can only isolate dissolved cytosm proteins at room temperature. More preferably, the particle size of the ultra fine silica powder is 600 to 800 탆.

Third, the alkali is added to the supernatant obtained above, and then hydrolyzed.

It is advisable to use sodium hydroxide (NaOH) as the alkali. This is because sodium hydroxide (NaOH) causes the supernatant to decompose into peptides having a small molecular weight.

NaOH is added so that the final concentration is 0.2-1 M, more preferably 1 M, and the mixture is heated at room temperature (15-30 ° C) for 0.5-1 hour, more preferably for 30 minutes to hydrolyze do. In other words, since the silkworm gland can be completely decomposed even at room temperature, it is possible to prevent the hydrolyzate obtained at the high temperature treatment from being partially deformed, and to separate the hydrolyzate without using a harmful solvent such as LiBr It is possible. The hydrolyzate means that the peptide is hydrolyzed in sericin and fibroin.

Fourth, hydrolysis after hydrolysis and neutralization and desalting are carried out to obtain a hydrolyzate derived from silkworm.

At this time, the neutralization process is carried out by adding an acid, preferably phosphoric acid, and neutralizing by adding until the pH is about 7.0 to 7.5.

The desalting process can be generally used as a conventional desalting method. However, in the case of a polymer having a molecular weight of several thousands or more, various methods using a difference in molecular size with a salt, such as gel filtration, ultrafiltration, and dialysis, are performed.

The hydrolyzate of the present invention thus obtained is excellent in the effect of adding serum-free serum, can be used as a serum substitute because of its excellent cell proliferation effect, and can also be used as a material having superior physical properties such as skin coating effect, moisturizing effect, antioxidant ability and skin affinity Do.

2. Preparation of cosmetic composition of the present invention

In the present invention, the hydrolyzate having excellent cell proliferation effect is dissolved to prepare a liquid state, and a cosmetic composition containing the hydrolyzate is prepared.

At this time, since the hydrolyzate is dissolved well in water, it is preferable to dissolve it in water.

The cosmetic composition preferably contains the hydrolyzate as an active ingredient singly, but economically, the hydrolyzate is contained in an amount of 0.001 to 50% by weight, preferably 0.1 to 15% by weight, based on the total weight of the cosmetic composition, More preferably 0.5 to 5% by weight, and the remaining balance may contain a material acceptable as other cosmetics.

At this time, when the hydrolyzate is contained in an amount of less than 0.001% by weight, the antioxidant ability, skin cell proliferating ability, skin affinity and the like are insufficient. When the hydrolyzate content is more than 50% by weight, the effect is equal to or less than 50% by weight.

The formulation of the cosmetic composition can be applied differently depending on the method and purpose of use.

Examples of the formulations include lotions, liquid preparations, emulsions, emulsions, suspensions, tablets, and capsules.

When used as cosmetics, it can be used as cosmetics such as foundation lotion, lipstick, mascara or makeup base such as soft lotion, milk lotion, nutrition cream, massage cream, essence, cleansing foam, cleansing water, pack or body oil And can be blended with components commonly used in external preparations for skin such as an aqueous component, an oily component, a surfactant, a moisturizing agent, a thickening agent, an antiseptic, a perfume pigment and the like.

The present invention will be described in more detail with reference to the following examples and experimental examples, but the scope of protection is not limited to the following examples and experimental examples.

Example 1: Hydrolysis product derived from silkworm gland

The silk was prepared with the raw material and lyophilized.

The lysosomes were separated from the lyophilized product, and the separated lysosomes were administered to a cryogenic mill to be processed into ultrafine powder.

The thus-prepared silkworm ultrafine powder was dissolved by stirring at room temperature (15 to 30 ° C) for 30 minutes. After centrifugation, the supernatant was taken, and NaOH was added to the supernatant. Lt; / RTI > for 30 minutes.

Then, the solution was neutralized to pH 7 with phosphoric acid, and desalted by ultrafiltration.

Thereafter, it was freeze-dried again to obtain the hydrolyzate used in the present invention.

<Example 2> Production of cream type cosmetics of the present invention

After dissolving the hydrolyzate of Example 1 in water, cosmetics having the following composition were prepared.

Hydrolyzate solution of Example 1 ------- 5.0 wt%

Cetanol 2.0 wt%

&Lt; tb &gt; &lt; tb &gt;

Sobutan monostearate ------- 1.0 wt%

Mineral oil -------------------- 10.0 wt%

Trioctanoate -------------- 5.0 wt%

Triethanolamine ---------------- 0.5 wt%

Carbomer ------------------------ 0.2 wt%

Glycerin 5.0 wt%

Propylene glycol - 3.0 wt%

Preservative, fragrance ---------------------

The remaining purified water

Example 3 Preparation of ointment-type cosmetics of the present invention

After dissolving the hydrolyzate of Example 1 in water, cosmetics having the following composition were prepared.

The hydrolyzate solution of Example 1 5.0 wt%

Diethyl sebacate 8.0 wt% &lt; RTI ID = 0.0 &gt;

Sintered ----------------------------------------- 5.0 wt%

Polyoxyethylene oleyl ether phosphate - 6.0 wt%

Sodium benzoate --------------------------------

The total amount of the remaining vaseline

Example 4: Preparation of an essence type cosmetic product of the present invention

After dissolving the hydrolyzate of Example 1 in water, cosmetics having the following composition were prepared.

The hydrolyzate solution of Example 1 5.0 wt%

Propylene glycol 10.0 wt%

Glycerin 10.0 wt%

Aqueous solution of sodium hyaluronate (1%) -------------- 5.0 wt%

Ethanol 5.0 wt%

Polyoxyethylene hydrogenated castor oil - 1.0 wt%

0.1% by weight &lt; RTI ID = 0.0 &gt;

Carbomer - 0.4 wt%

Incense ---------------------------------------

The remaining purified water

<Experimental Example 1> Antioxidant ability analysis by hydrolyzate separation process

The DPPH test was conducted to measure the antioxidant activity of the hydrolyzate of Example 1, and the degree of antioxidant activity with the vitamin C solution was compared as a control.

The results are shown in Table 1 below.

Antioxidant activity (%) Frequency Average One 2 3 15% vitamin c 97 97 97 97 The hydrolyzate of Example 1 82 82 81 84

As shown in Table 1, the hydrolyzate of Example 1 showed similar results to the control group.

In other words, vitamin C is one of the trace elements for maintaining the function and health of the human body. It helps the human body to resist infection, heals the wounds and keeps the tissues healthy, and as one of the antioxidants, It is a known ingredient.

Thus, the present invention has a similar antioxidant activity to that of the control group.

As shown in FIG. 1, the antioxidative ability of the extract of the present invention was higher than that of extracts of silkworm gland secreted with lithium bromide (LiBr) and urea (organic compound of CO (NH ₂ ) ₂ ) 1 hydrolyzate showed the highest antioxidant activity.

<Experimental Example 2> Cytotoxicity test of hydrolysates

To investigate the cytotoxicity of hydrolysates, stem cells were cultured and hydrolysates were added and cytotoxicity was confirmed by CCK-8 assay.

The CCK-8 assay can be used to determine whether a tetrazolium salt that has been treated in cell culture has been transformed into a dark red-colored formazan by the mitochondrial succinate-tetrazolium-reductase system (formazan), and the amount of viable cells can be quantitatively measured by measuring at 450 nm.

The hydrolyzate at a concentration of 0 to 5 mg / ml based on the IC ₅₀ concentration was treated on the cultured cells and CCK-8 assay was performed after further culturing for 2 days.

As a result, as shown in Fig. 2, it was confirmed that the hydrolyzate of Example 1 of the present invention was not toxic because no inhibition of cell proliferation occurred.

Experimental Example 3 Measurement of proliferation effect of human dermal fibroblast using hydrolyzate

The hydrolyzate of Example 1 was dissolved in DPBS (WelGENE, cat. LB 001-02) at a concentration of 20 mg / ml, and then treated with the concentrations of 0 mg / ml and 0.1 mg / ml in the serum-free medium.

To compare growth with 10% serum added cells (10% FBS), WST-1 assay (Daeillab) was measured for 4 days at intervals of one day.

The WST-1 assay reagent was added at 10% of the culture medium, incubated for about 3 hours in the same incubation condition, and absorbance was measured at 450 nm.

As a result, as shown in FIG. 3, when the 1 mg / ml silkworm-derived hydrolyzate was added to the serum-free culture solution, the cell proliferation effect was superior to that of the other company's cocoon-derived sericin of the same concentration. The hydrolyzate derived from the sebaceous glands prepared in the present invention has an excellent effect on the proliferation of human skin cells.

<Experimental Example 4> Cell adhesion test of hydrolysates

The hydrolyzate prepared in Example 1 was dissolved in DPBS (WelGENE, Cat. LB 001-02) at a concentration of 0.2, 0.5 and 1.0 mg / ml, and then treated on the surface of a culture dish (SPL, 10035) Lt; / RTI &gt; incubator.

Then, the hydrolyzate was removed, washed twice with phosphate buffer solution and completely dried at room temperature

Cell adhesion test was carried out for 2 days after cell inoculation by the coating concentration of the hydrolyzate, and the number of cells proliferated was directly measured.

As a result, as shown in FIG. 4a, it was confirmed that the cells were adhered to the culture dish with increasing amount of the hydrolyzate after 30 minutes of inoculation.

As can be seen from FIG. 4B, the total number of cells proliferated after 2 days of culture showed that the higher the concentration of the coated hydrolyzate, the greater the amount of proliferated cells.

Therefore, the hydrolyzate obtained in the present invention not only serves as an additive for promoting cell proliferation, but also has the effect of promoting proliferation of cells when coated on a culture dish.

<Experimental Example 5> Skin irritation test of hydrolysates

The test was carried out in accordance with the "Toxicological Test Standards for Drugs, etc." notified by the Food and Drug Administration.

To investigate the skin irritation of the hydrolyzate of Example 1, 1% (w / v) hydrolyzate of Example 1 was applied to the normal skin of the New Zealand male white rabbit The non - treated normal skin (non - irritation and skin) and the irritated skin were compared and evaluated.

Twenty-four hours before the test, healthy epidermal skin was prepared using an electric epilator to prevent skin damage on the dorsal skin of the rabbit, and the skin was divided into left and right sides to prepare four areas of 2.5 cm × 2.5 cm.

The hydrolysates of Example 1 were sufficiently applied to healthy skin and cheek skin, and scored according to skin reaction standards at 24, 48 and 72 hours, and recorded.

As a result, as shown in FIG. 5, in the evaluation of the final irritation, the hydrolyzate of Example 1 of the present invention was evaluated as non-magnetic (evaluation index 0 to 0.5), and the hydrolyzate was found to be a material free from skin irritation.

As described above, according to the present invention, it is possible to provide a cosmetic composition using a hydrolyzate derived from silkworm-derived hydrolyzate having excellent antioxidative ability, skin cell proliferating ability, skin-affinity and the like without harming human body by being able to obtain hydrolyzate by room temperature extraction without using harmful solvent .

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. You can implement the examples. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.

Claims (8)

Cosmetic composition containing a silkworm-derived hydrolyzate
The method according to claim 1,
The hydrolyzate is obtained from the silkworm by separating the silk glands from silkworms, dissolving the isolated glands with water, and then adding the alkali to the supernatant obtained by centrifugation, followed by hydrolysis and neutralization and desalting.
Cosmetic composition.
The method according to claim 1,
The hydrolyzate comprises a peptide hydrolyzed by sericin and fibroin,
Cosmetic composition.
The method according to claim 1,
The hydrolyzate is characterized in that it contains 0.001 to 50% by weight based on the weight of the cosmetic composition,
Cosmetic composition.
The method according to claim 2,
The hydrolysis is characterized in that made at 15 ~ 30 ℃,
Cosmetic composition.
The method according to claim 2,
Using NaOH as the alkali,
Cosmetic composition.
The method according to claim 2,
When neutralizing using phosphoric acid,
Cosmetic composition.
The cosmetic composition according to any one of claims 1 to 7, which has a formulation of any one of lotion, essence, lotion, cream, pack, foundation, gel, ointment or spray,
Cosmetic composition.

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