KR20170061509A - Cosmetic composition containing egg yolk having antibody for prevention or improvement of athlete's foot - Google Patents

Abstract

The present invention relates to a cosmetic composition for prevention or improvement of athlete's foot comprising an egg yolk-containing antibody against an athlete's fungus. More specifically, the present invention relates to a cosmetic composition for preventing or improving athlete's foot comprising an egg yolk containing IgY against tricophyton rubrum (KCTC No. 6375, Trichophyton rubrum ) To a cosmetic composition for prevention or improvement of athlete's foot.
The cosmetic composition for prevention or improvement of athlete's foot according to the present invention contains egg yolk having a specific antibody against tricofitone rubrum, which is a fungus bacterium, so that athlete's foot can be effectively prevented and improved, Can be reduced.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a cosmetic composition for prevention or improvement of athlete's foot comprising an egg yolk-

The present invention relates to a cosmetic composition for preventing or improving athlete's foot, and more particularly, to a cosmetic composition for preventing or improving athlete's foot containing an egg yolk specific antibody for athlete's foot.

Dermatophytosis is an infection caused by dermatophytes which are classified as fungus. According to the site of occurrence, it is classified into tofu tenderness, body tenderness, cysts, facial tenderness, tender tenderness, Among them, the most common infectious disease is called athlete's foot, which accounts for 35% to 40% of the total.

Generally, dermatomyositis refers to fungi that live on the keratin of the skin and grow in a thread shape. It is a kind of fungus that contains mold, mushroom, yeast, etc. It melts the skin's keratin to make nutrients. It is a disease caused by foot, rarely parasitic and breeding on the skin of hair, and it occurs in the area where the keratin is abundant, moist and warm. The reason these molds cause problems on the feet is because the shoes provide a warm, humid environment that fungus likes.

There are three types of symptoms of athlete's foot. The first type is a rash type, with blisters on the instep or on the soles, and most of the time it is accompanied by itching. The blisters burst, the secretions come out, and the skin becomes red. The second type occurs between the toes. The thighs are swollen and become whitish. Initially, the skin is a little peeled off, but if you do not take care of the secretion. If it is characterized by itching, it may swell around the wound and cause pain and pain, and the limbs and lymph glands may swell, causing the legs to slough. Finally, there is an exocrine proliferative type, which does not have much secretion but it is accompanied by itching, thickening of the soles, redness and peeling of the skin. The mold causing this symptom is easy to infiltrate the toenails and is often caused by the combination of the ringworm.

It is known that there are 43 species belonging to three genus of Trichophyton (T), Microsporum (M) and Epidermophyton (E) in the world, 10 species are found. Among them, Trichophyton rubrum is the causative organism found in more than 80% of athlete's foot patients. In addition, T. mentagrophytes or E. floccosum may be the cause.

The cause of skin fungal growth in skin is the outermost layer of skin that protects the body from harmful external factors, composed of dead keratinous dead cells. Most bacteria do not pass through the stratum corneum, It has enzymes that can break down (Keratinase), destroys the stratum corneum, uses it as nutrients, and lives on the skin parasitically. On the other hand, fungi are less resistant to human immunity than germs, so infection to the human body is mostly restricted to the skin. However, when the fungus penetrates and weakens the stratum corneum, other bacteria may infiltrate and cause inflammation (secondary bacterial infection), which causes the athlete's foot to fail.

In the case of a polyclonal antibody to be compared with an egg yolk antibody, a method of producing an antibody is a conventional method in which a vaccine containing a desired antigen is injected into a mammal such as a rabbit, a horse, a goat, a sheep, a dog, And then the blood of the animal is extracted from the serum. This method is not easy to obtain a large amount of blood, and therefore, it is difficult to expand to an industrial scale. On the other hand, the egg yolk antibody method is advantageous in that it can easily obtain a large amount of antibody only by egg yolk separation without an effort such as blood sampling by immunizing a chicken.

In the case of mammals, the immune function of the offspring is handled until the active immune function, which is able to produce and defend itself by transferring the immune antibody to the offspring through the placenta or breast milk, is matured. However, in the case of birds, blood antibodies are accumulated in egg yolk and egg whites in order to transmit the immune ability possessed by the hen to the offspring. IgA, which is responsible for innate immunity (passive immunization), immune antibody IgY is enriched in egg yolk. Therefore, when egg yolk is separated and purified to produce IgY which plays a similar role to IgG in mammalian blood, it is possible to produce 50 to 100 mg of IgY with 1 egg and 5 to 7 eggs per week, Month, it is possible to produce egg yolk antibody of 1.5g efficiently. In addition, the egg yolk antibody has a molecular weight of about 200 to 220 kDa, which is larger than that of mammalian IgG and has advantages over mammalian antibodies.

As a method for treating athlete's foot, there is a method of using acetic acid or a derivative thereof which has a strong inhibitory effect against reproductive fungi, and a method of using antioxidant activity such as resorcinol, fluo, and benzoyl peroxide Suppression method, and the like. Although these methods have achieved some results in the prevention and treatment of athlete's foot, they still have many problems in terms of skin safety such as irritation of skin, concern of contact dermatitis, damage of skin tissue itself, and destruction.

On the other hand, the prior art related to a cosmetic composition for treating athlete's foot is as follows.

Korean Patent Registration No. 10-0768401 (patent holder: Dr. SHOP Co., Ltd., filed on March 22, 2001) relates to a cosmetic composition for preventing athlete's foot, more specifically, a composition containing urethropropyl butylcarbamate Which is capable of inhibiting athlete's foot by selectively inhibiting the growth of athlete's foot. The urethropropyl butylcarbamate according to the present invention has excellent antimicrobial activity against acebacteria, and a cosmetic composition such as a cosmetic composition containing the same exhibits the most excellent effect as compared with the conventional antimicrobial agent.

Korean Patent Registration No. 10-0553330 (patentee: Kolmar Co., Ltd., filed on March 20, 2003) discloses a citrus essential oil having antimicrobial activity against a causative agent of athrochinoid and a cosmetic composition for prevention of athlete's foot comprising the same. Specifically, an essential oil component having antimicrobial activity against Trichophyton rubrum and Trichophyton mentagrophytes, which are major pathogens causing skin diseases such as athlete's foot, is efficiently extracted from citrus unshiu The present invention relates to a cosmetic composition for preventing and treating athlete's foot, which contains citrus essential oil without side effects and is a component isolated from natural products. The citrus essential oil extracted and extracted from citrus fruits according to the present invention and the cosmetic composition containing the citrus essential oil have excellent antimicrobial activity against tricofitone rubrum or tricophyton mentagrophite, It can be usefully used in a composition.

However, in the prior art, there is no teaching or research on a cosmetic composition for prevention or improvement of athlete's foot containing egg yolk containing antibody IgY against acebacteria as in the present invention.

In order to achieve the above object, the inventors of the present invention found that egg yolk containing antibody IgY against fungal athletes has antimicrobial activity against causative agents of athymic fungi by measuring antimicrobial activity against tricofitone rubrum Thus completing the present invention.

Accordingly, the present invention provides an antimicrobial activity against tricofitone rubrum, which is a major causative agent of a skin disease such as athlete's foot, by using it in an egg yolk having an antibody against acebobotia and a cosmetic composition containing it, It is an object of the present invention to provide a cosmetic composition for prevention or improvement of athlete's foot which does not have a sense of resistance and does not cause antibiotic resistance and side effects.

The present invention relates to tricophyton rubrum (KCTC No. 6375, Trichophyton The present invention provides a cosmetic composition for prevention or improvement of athlete's foot comprising an egg yolk containing IgY against rubrum as an active ingredient.

Preferably, the active ingredient of the present invention is 0.10 to 20% by weight based on the total weight of the cosmetic composition for improving athlete's foot.

Preferably, the composition of the present invention is manufactured by a formulation selected from the form of a hand and foot cream, a gel, a lotion, a skin, a tissue, a lotion and a cleanser formulation.

As described below, the cosmetic composition for prevention or improvement of athlete's foot according to the present invention contains egg yolk having a specific antibody against tricofitone rubrum, which is a fungal anomalous bacterium, so that athlete's foot can be effectively prevented and improved , And by using natural materials, it is possible to reduce consumers' rejection.

Fig. 1 shows the results of confirming the content of IgY contained in egg yolk obtained from immunized laying hens by inoculating vaccine-1 with a protein chip.
FIG. 2 shows the results of analysis using a vaccine prepared by the vaccine production method-2 of Example 1 of the present invention.
FIG. 3 shows the results of confirming whether egg yolk antibodies were produced using egg yolks obtained from immunized laying hens by inoculating the vaccine-3 prepared in step 2 of Example 1 of the present invention.

The present invention relates to tricophyton rubrum (KCTC No. 6375, Trichophyton The present invention provides a cosmetic composition for prevention or improvement of athlete's foot comprising an egg yolk containing IgY against rubrum as an active ingredient.

Hereinafter, the present invention will be described in detail.

The cosmetic composition of the present invention contains, as an active ingredient, an anabolic agent, more specifically, tricofitone rubrum (KCTC No. 6375, Trichophyton and egg yolk containing IgY to rubrum .

Egg yolk lutea is a form of passive immunization using the avian immune system. In the case of ovaries below algae, the immune antibody obtained by active immunization of the mother chicken is transferred to the egg yolk and immunity is transmitted to the offspring. The IgGdhk-like IgY in the blood is the most abundant of the antibodies in egg yolk. It has a molecular weight of about 200 to 220 kDa, which is larger than that of mammalian IgG and has advantages over mammalian antibodies.

The yolk of the present invention can be produced by the following method. First, athymobacteria, more specifically tricofitone rubrum (KCTC No. 6375, Trichophyton rubrum ) is cultured and diluted to prepare a cultured strain solution. In order to disrupt the athlete's fungus, the cultured strain solution is treated with an ultrasonic cooker, lyophilized, the lyophilized strain is pulverized with liquid nitrogen, the resulting strain is mixed with the adjuvant at a certain ratio, Thereafter, the vaccine can be prepared by emulsification using a homomixer. By performing the above-described disruption, the protein can be prevented from being denatured, thereby maintaining the role of the vaccine. The vaccine thus obtained is inoculated into layer chicken, and if necessary, can be inoculated several times over a certain period of time. Eggs obtained from these laying hens are separated from egg yolk and lyophilized to obtain egg yolk powder.

With regard to the disruption process of athlete's fungi, fungi is a class of eukaryotes including yeast, fungi, and mushroom as fungi, and is an independent system distinguishable from animals, plants, and bacteria, and is a member of various cells including nucleus and mitochondria Since the organelle is a multicellular organism composed of chitin and the mycelium wall is composed of chitin, it is impossible to destroy the mycelial wall of athlete's fungus by disrupting the cell membrane of a bacterium, which is a single cell organism and a prokaryote, It seems that the immune response to several proteins did not occur. Although there are various experimental methods for crushing aphthous fungi known in the prior art, buffer solutions that do not have harmful effects when the buffer solution is injected into the laying hens should be used, and methods of chemically treating such as high temperature, Is modified and can not serve as a vaccine, it is preferable to use a method of pulverizing by applying a physical force in order to produce the vaccine. Accordingly, in the present invention, a method of pulverizing plants or animal tissues by using a mortar bowl after being frozen with liquid nitrogen, which is a method of pulverizing plants or animal tissues, was used.

The egg yolk containing the vaccine obtained in the present invention exhibited a tendency to increase in the amount of yolk antibody when the antibody titer was measured and the number of weeks of antibody vaccination was increased. In the 5th week, (See FIG. 3). Accordingly, the cosmetic composition for prevention or improvement of athlete's foot according to the present invention contains egg yolk having an antibody specific for athlete's foot obtained through such an antibody, so that athlete's foot can be effectively prevented and improved, Can be reduced.

The cosmetic composition of the present invention preferably contains 0.10 to 30% by weight, preferably 0.10 to 20% by weight, based on the total weight of the cosmetic composition for the improvement of athlete's foot, of egg yolk containing IgY against fungal athletes of the present invention. If the amount is less than the above range, there is a disadvantage that the antimicrobial activity against anthracobacteria is insufficient, and if it exceeds the above range, problems may arise in the form.

In addition to the egg yolk, the cosmetic composition of the present invention may further contain a natural or synthetic substance exhibiting activity against acebacteria known in the art, and the content thereof may be appropriately controlled. Such additional materials and contents are not particularly limited and may be appropriately selected and adjusted according to knowledge in the field.

The cosmetic composition of the present invention can be manufactured into cosmetic formulations that can be used in athlete's foot. Preferably, the cosmetic composition can be manufactured into a form selected from the form of hand and foot cream, gel, lotion, skin, tissues, lotion and cleanser. Methods and components for the preparation of each of these formulations are well known in the art and are not specifically described herein.

Hereinafter, the structure and function of the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following examples, but includes modifications of equivalent technical ideas.

Example  One: Athlete's foot  Antibody against IgY  Egg yolk production

Example 1 produced egg yolk containing IgY against Trichophyton rubrum .

Step 1: Antigen production

Trichophyton rubrum (KCTC No. 6375, Trichophyton rubrum) was cultured in a Sabourauds Agar medium for 3 to 4 days at 37 ° C in an incubator. After confirming the colony, the cells were inoculated into the prepared medium and cultured in a shaking incubator at 25 ° C for 10 days. The obtained culture was inactivated with 0.2% formalin, centrifuged at 20000 x g for 15 minutes, and then the supernatant was removed. The resulting precipitate was again inactivated with 1X PBS, 0.1% formalin buffer solution. Then, the sample was diluted to a OD value of 0.5 at a wavelength of 400 nm using a spectrophotometer.

Step 2: Preparation of vaccine

Using the strain solution cultured in step 1, a vaccine was prepared by the following three vaccine production methods.

Step 2-1: Vaccine production method-1

The strains cultured in step 1 and Adjuvant (ISA 70 VG, Montanide, Seppic) were mixed at a ratio of 3: 7 and then emulsified for 5 minutes at 3000 rpm using a homo-mixer. 1.

Step 2-2: Vaccine Manufacturing Method-2

The strain solution cultured in step 1 was repeated 10 times with a pulse of 30 seconds at 70% intensity and a stop of 20 seconds using an ultra-sonicator. This solution and Adjuvant (ISA 70 VG, Montanide, Seppic) were mixed at a ratio of 3: 7 and emulsified at 3000 rpm for 5 minutes using a homo-mixer to prepare vaccine-2 [ 2].

Step 2-3: Vaccine Manufacturing Method 3

The solution of the strain cultured in step 1 was repeated 30 times for 30 seconds at 70% intensity using an ultrasonic cleaner and for 10 seconds for 20 seconds. These solutions were frozen at -83 ° C and lyophilized in a freeze dryer for 24 hours. The lyophilized strains were collected and transferred to a mortar, and then pulverized with a mortar while pouring liquid nitrogen (-196 ° C) little by little. The mixture was pulverized about 10 times until the aggregated mass was not visible. The mixture was mixed with Adjuvant (ISA 70 VG, Montanide, Seppic) at a ratio of 3: 7 and homomixed with a homo-mixer And then emulsified at 3000 rpm for 5 minutes to prepare vaccine-3.

Step 3: Production of yolk with a specific antibody against acebacteria

The vaccine-1 prepared in step 2 was inoculated into the breast muscle at 0.5 ml in a 19-week-old laying hens and then inoculated two weeks later.

Egg yolk was separated using a commercially available egg yolk separator, and after separation and freeze drying, the egg yolk powder was added to the mill to produce egg yolk powder.

< Experimental Example  1 > of a fungal specific antibody in egg yolk Potency  Measure

 Reverse transcription of anthracoblast-specific antibody in egg yolk was measured by replacing the plate on which the experiment was performed with a protein chip in a 96-well plate in the same manner as the 'Indirect ELISA Method'.

Egg yolks collected for 1 week before inoculation were used as a control group. Egg yolks collected for 1 week (total 4 weeks) after the first inoculation were measured by week (control, 1, 2, 3).

The antigen was coated on a protein chip having 96 wells at a concentration of 200ng / ml in 1 占 퐇, and left overnight at 4 占 폚 in a wet box. After washing with PBS-T (phosphate buffer saline, 0.5% Tween-20, pH 7.4), the cells were blocked with PBS buffer containing 1% BSA in a shaking incubator at room temperature for 1 hour. Washed with tertiary distilled water and dried with nitrogen gas. The negative control and sample (week 3) were diluted 1/10 with blocking buffer, and 1 당 / well was added and allowed to stand at room temperature for 1 hour. (Anti-Chicken IgY, Alexa-635 labeled) diluted 1/100 was reacted for 1 hour in the same manner as the sample treatment, washed and dried in the same manner, and the resultant was analyzed with a fluorescence microarray scanner (Genepix4000B, Axon) was used to measure the degree of binding of a fungal specific antibody contained in each well.

The results of measuring the activity of the IgY antibody in egg yolk obtained from immunized laying hens by inoculation with vaccine-1 are shown in FIG. Fig. 1 shows the result of confirming the content of IgY contained in egg yolk obtained from immunized laying hens by inoculation with vaccine-1 with a protein chip. As can be seen from FIG. 1, the acyclovir exhibited more than 7-fold increase in the IgY antibody titer of egg yolk after 3 weeks of immunization compared to the control group, but the production of specific antibody was not observed in the case of anthracobacteria I did.

Vaccine-2 was prepared by using a stronger ultrasonication method on the assumption that the immune reaction did not occur due to the problem of cell lysis during the production of vaccine, but no specific antibody was produced. More specifically, FIG. 2 shows the result of analysis using the vaccine prepared by the above-mentioned vaccine production method-2. In order to increase the concentration of athlete's antigens above the method shown in FIG. 1, The results were obtained by treating the yolk antibody of the control group on the left three columns and the egg yolk antibody of the nasophilus on the right three columns, respectively. As the concentration of the antigen decreases, the degree of binding of the yolk antibody of anthracnose appears to be higher than that of the control, but the yolk antibody does not appear to have actually occurred because it is within the standard deviation.

In addition, 0.5 ml of the vaccine-3 prepared in step 2 was inoculated into the chest muscles and then inoculated two weeks later. Three weeks after the second vaccination, egg yolks were collected for up to 5 weeks. The result of measuring the activity of the antibody using the antibody was shown in FIG.

FIG. 3 shows the results of confirming the production of egg yolk antibody using the yolk obtained from the immunized laying hens by inoculating the vaccine-3 prepared in the step 2 above. After treatment with the same concentration of athlete's antigens, the egg yolk antibody collected weekly for each row was examined. As the number of weeks increased, the egg yolk antibody tended to increase gradually. In the 5th week, It was confirmed that the antibody was generated.

Formulation Example 1: Preparation of gel containing yolk powder

To 5 g of 95% ethanol were added 2.5 g of propylene glycol, 2.5 g of glycerin, 0.05 g of allantoin, 0.05 g of disodium iodide, 0.6 g of Fizi-40 hydrogenated castor oil, 0.2 g of menthol, 0.2 g of methylparaben, Were mixed and dissolved. 5 g of the egg yolk powder obtained in Example 1 and 0.8 g of Carbomer 940 were dissolved in 77.9 g of purified water. The mixture was added to the mixture and stirred. To the 4.3 g of purified water, 0.8 g of triethanolamine was added and dissolved. .

Formulation Example 2: Preparation of cream containing yolk powder

1.5 g of stearyl alcohol, 1 g of beeswax, 5 g of vaseline, 1 g of glyceryl monostearate, 0.8 g of sorbitan stearate, 2 g of polysorbate 60, 37 g of mineral oil, 5 g of macadamia nut oil, 0.3 g of silicone oil, g was mixed with heat at 70 to dissolve, and 5 g of egg yolk powder obtained in Example 1, 0.2 g of methylparaben, 0.15 g of carbomer-940, 5 g of glycerin, 0.07 g of allantoin, 0.03 g of disodium iodide, , And 35.7 g of purified water were heated to 75 to dissolve. The mixture was emulsified by mixing them and then cooled to prepare oil-in-water (O / W) type emulsion.

Claims (4)

Trichophyton rubrum (KCTC No. 6375, Trichophyton The present invention relates to a cosmetic composition for preventing or improving athlete's foot comprising an egg yolk containing IgY against rubrum as an active ingredient.
The cosmetic composition for preventing or improving athlete's foot according to claim 1, wherein the effective ingredient is 0.10 to 20% by weight based on the total weight of the cosmetic composition for improving athlete's foot.
The cosmetic composition for preventing or improving athlete's foot according to claim 1, wherein the composition is prepared from a form selected from the group consisting of hand cream, gel, lotion, skin, tissue, lotion and cleanser.

The method according to claim 1, wherein the yolk sac is cultured and diluted to prepare a cultured strain solution, and then the cultured strain solution is treated with an ultrasonicizer, lyophilized, and the lyophilized strain is immersed in a liquid The resulting vaccine was inoculated into a layer chicken, and an egg obtained from this layering system was inoculated with the vaccine. The obtained vaccine was inoculated into a layer chicken, and the resulting egg was mixed with an adjuvant, followed by emulsification using a homomixer to prepare a vaccine. And the yolk powder is obtained by lyophilizing the yolk powder. The cosmetic composition for prevention or improvement of athlete's foot.

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