KR101140573B1 - External Skin Preparation Using an Extract of Horse Plancenta - Google Patents

Abstract

The present invention relates to an external composition for skin using horse placenta extract. Specifically, the present invention relates to an external composition for skin having skin whitening, skin moisturizing and atopic dermatitis improving activity using horse placenta extract.

Horse Placenta, Skin, Whitening, Moisturizing, Atopy

Description

External skin preparation using an extract of horse plancenta

The present invention relates to an external composition for skin using horse placenta extract. Specifically, the present invention relates to an external composition for skin having skin whitening, skin moisturizing and atopic dermatitis improving activity using horse placenta extract.

Human skin is changed by internal and external factors. Internally, the elasticity decreases due to a decrease in the activity of skin cells, a decrease in skin immunity, externally by ultraviolet rays, environmental pollutants, etc., and the aging of the skin progresses due to the occurrence of spots, freckles, and mushrooms.

In particular, atopic dermatitis caused by disturbance of the skin immune system is widespread due to the use of automobile exhaust gas and various chemicals whose toxicity is not precisely identified, and the age of onset continues to decrease. This phenomenon is further accelerated by changes in diets focused on instant food and meat.

On the other hand, since the placenta is a tissue responsible for material exchange between the fetus and the mother and contains various bioactive substances necessary for the growth of the fetus, there have been studies on the efficacy of the placenta, for example, the treatment effect of acquired vitiligo and antioxidants. Activity, cell regeneration and the like are known as the placenta's efficacy.

The present invention has been completed by confirming that horse placenta extract and the like have activity against skin diseases such as blemishes, freckles, and atopic dermatitis.

Accordingly, an object of the present invention is to provide an external composition for skin having skin disease improvement activity such as blemishes, freckles, atopic dermatitis using horse placenta extract and the like.

Specific objects or aspects of the present invention will be presented below.

In one aspect, the present invention relates to a skin external preparation composition for skin whitening.

The inventors of the present invention, as confirmed in the following Examples and Experimental Examples, when the horse placenta extract and / or Moonjuran leaf extract was administered to the mouse melanoma cells B16F10 together with α-MSH, a melanogenesis-stimulating agent, all of the above extracts melanin production It could be confirmed that suppressing.

Therefore, the skin external preparation composition for skin whitening of the present invention is characterized in that it comprises a horse placenta extract and / or Moonju egg extract as an active ingredient.

In the present specification, the "horse placenta extract" refers to an extract obtained by any extraction method and extracts the horse placenta (especially the chorionic tissue from which the amniotic membrane and the umbilical cord are removed from the horse placenta), and preferably, An extract obtained by the extraction method used in the example, that is, an extract obtained by acid hydrolysis, an extract obtained by decomposition with protease or an extraction solvent, in particular, an extract obtained by immersion in water, ethanol or a mixed solvent thereof. Such an extract may be in a purified form by filtration, in the case of an extract obtained by acid hydrolysis, in a neutralized form by a base, or in a concentrated form or a solid form in which an extraction solvent or the like is removed.

In addition, in the present specification, the "moonjuran leaf extract" means an extract obtained by using the moonjuran ( Crinum asiaticum ) leaf as an extraction target and using water, ethanol or a mixed solvent thereof. Munjuran leaf extract may also be in a purified form or a concentrated form or solid form in which the extraction solvent is removed. The leaves of Munjuran have been known to have effects such as analgesic, detoxification, and seedling.

Also herein, "skin whitening" can be understood as a result of the inhibition of melanin production. Specifically, as the production of melanin is inhibited, the symptoms resulting from the increase of melanin (when the composition for skin whitening of the present invention is identified as a pharmaceutical composition may be regarded as a disease) are prevented or suppressed. It is understood as a result. Such symptoms may include blemishes, freckles, blotch, skin aging, etc., but the horse placenta extract and / or moonshine egg leaf extract included in the skin whitening composition of the present invention as shown in the following Examples and Experimental Examples of the present invention. As long as it is evident to show an effect that inhibits the production of melanin, the symptom is a symptom resulting from an increase in melanin and should be understood as all symptoms leading to skin whitening by preventing or inhibiting it.

In addition, in the present specification, the "active ingredient" alone means a component that can exhibit the desired activity or the activity itself with a carrier that is not active.

In the present invention, the following Examples and Experimental Examples show that the effect of inhibiting melanogenesis is increased when the horse placenta extract is used in combination with the Moonjuran extract. Therefore, it is preferable that the skin external preparation composition for skin whitening of the present invention includes both the horse placenta extract and the moonju egg extract which are the active ingredients.

The skin external preparation composition for skin whitening of the present invention may include a compound or a plant extract that is known to have a skin whitening effect so as to enhance or enhance the skin whitening effect, in addition to the horse's placenta extract and / or moonshine egg leaf extract, which are active ingredients, It may also include other known compounds or plant extracts known to have a skin whitening effect, and may further include compounds or plant extracts having a skin whitening effect that will be revealed in the future.

Herein, the other compounds or plant extracts include mercaptosuccinic acid, mercaptodextran, tefrenone, dihydroxy-isoquinoline, indomethacin, 3-hydroxymanul, vitamin K, thiazolidone, kynurenine, and lemon. Extract, cucumber extract, mulberry extract, licorice extract, rosemary extract, acerola cherry extract, ginkgo extract, carob (carob) extract, geranium extract, herbal extract, and the like, but is not limited thereto.

Compounds or plant extracts as exemplified above may be included in the skin whitening composition for skin whitening of the present invention together with one or more horse placenta extracts and / or moonshine eggs extract.

On the other hand, in the skin external preparation composition for skin whitening of the present invention, the horse placenta extract and / or moon julan leaf extract as an active ingredient does not have toxicity to the human body depending on the use, formulation (cosmetic, soap, ointment, etc.). It may be included in any amount within, specifically, may be included from 0.05% to 99.99% by weight relative to the total weight of the composition.

In another aspect, the present invention relates to a skin external preparation composition for moisturizing the skin.

The following Examples and Experimental Examples show the skin moisturizing effect of horse placenta extract and / or moonjuran leaf extract as well as the skin whitening activity, especially when both horse placenta extract and moonjuran leaf extract are used. Is showing.

Therefore, the skin external preparation composition for moisturizing the skin of the present invention is characterized in that it comprises a horse placenta extract or moonju egg extract, preferably characterized in that it comprises all of the above extract.

In the skin moisturizing composition for skin moisturizing of the present invention, the meaning of the horse placenta extract, the meaning of the munjoran leaf extract, the meaning of the active ingredient, the content of these active ingredients, etc. are as described above for the skin whitening composition for skin whitening of the present invention. It is still valid.

In another aspect, the present invention relates to a composition for external application for skin to improve atopic dermatitis.

Based on the following Examples and Experimental Examples, the composition for improving atopic dermatitis of the present invention is characterized in that it comprises a horse placenta extract or moonju egg extract, preferably comprises all of the above extract .

In the topical dermatitis composition for improving atopic dermatitis of the present invention, the meaning of the horse placenta extract, the meaning of the leaf extract extract, the meaning of the active ingredient, the content of these active ingredients, etc. The description is valid as it is.

The composition of the present invention (collectively, the skin external preparation composition for skin whitening, the skin external preparation composition for moisturizing the skin and the atopic dermatitis composition) may be regarded as a cosmetic composition in a specific embodiment.

When the composition of the present invention is conceived as a cosmetic composition, the cosmetic composition can be prepared in various forms, for example, emulsions, lotions, creams (oil-in-water, water-in-oil, multiphase), solutions, suspensions (anhydrous and Aqueous), anhydrous products (oil and glycol based), gels, masks, packs or powders and the like.

The composition of the present invention may include an acceptable carrier in cosmetic preparations in addition to the horse placenta extract and / or moonshine egg leaf extract as its active ingredient.

Herein, "acceptable carriers in cosmetic preparations" are compounds or compositions already known and used that may be included in cosmetic preparations, or compounds or compositions which will be developed in the future, which have no toxicity, instability or irritation more than the body can adapt to in contact with the skin. Say that.

The carrier may be included in the cosmetic composition for skin whitening of the present invention at about 1% by weight to about 99.99% by weight, preferably about 50% to about 99% by weight of the composition.

However, since the ratio depends on the form of the above-described preparation of the cosmetic of the present invention and its specific application site (face or hand) or its preferred application amount, the ratio is in any aspect the scope of the present invention. It should not be understood as limiting.

Meanwhile, examples of the carrier include alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscosity modifiers, emulsions, stabilizers, sunscreen agents, colorants, fragrances, and the like.

Alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, sunscreens, colorants, compounds / compositions that can be used as fragrances, etc., which can be used as the carrier are already known in the art. As a result, those skilled in the art can select and use appropriate materials / compositions.

On the other hand, the composition of the present invention (particularly, a topical dermatitis composition for improving atopic dermatitis) may be regarded as a pharmaceutical composition in another specific embodiment.

The pharmaceutical composition of the present invention is administered directly to the skin in a topical formulation.

In addition, the pharmaceutical composition of the present invention can be formulated using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants commonly used in the formulation, creams, lotions, ointments (semi Solid external preparations), microroemulsions, gels, pastes, transdermal agents (TTS) (such as patches, bandages, etc.) and the like.

The daily dosage of the pharmaceutical composition of the present invention is usually 0.001 ~ 150 mg / kg body weight range, it can be administered once or divided into several times. However, since the actual dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, and the severity of the patient, the dosage is in any aspect the scope of the present invention. It should not be understood as limiting.

The composition of the present invention may be regarded as a soap composition in another specific embodiment.

When the composition of the present invention is understood as a soap composition, the soap composition of the present invention may be prepared by including a horse placenta extract and / or moonjulan extract, which are active ingredients on a soap base, and as an additive, a skin moisturizer, an emulsifier, a hard water softener. And the like can be prepared.

As the soap base material, vegetable oils such as palm oil, palm oil, soybean oil, perm oil, olive oil, palm kernels, or animal oils such as tallow, pork, sunny and fish oils can be used, and the skin moisturizing agent is glycerin, erythritol and polyethylene glycol. , Propylene glycol, butylene glycol, pentylene glycol, hexyl glycol, isopropyl myristate, silicone derivatives, aloe vera, sorbitol and the like can be used, the emulsifiers include natural oils, wax fatty alcohols, hydrocarbons, natural plant extracts, etc. May be used, and tetrasodium idieti may be used as the water softener.

The soap composition of the present invention may further include an antimicrobial agent, a dispersant, a foam inhibitor, a solvent, a scale inhibitor, a corrosion inhibitor, a perfume, a pigment, a metal ion blocking agent, an antioxidant, a preservative, and the like as an additive.

In the soap composition of the present invention, the soap base or additives may be included in an amount generally used in the art, and the soap base is generally 99.9 to 50 wt% based on the total content of the soap composition. It may be added, the additive may be added in 1% to 20% by weight.

According to the present invention, it is possible to provide an external composition for skin having a skin whitening activity, a skin moisturizing activity and an atopic dermatitis improving activity using a horse placenta extract and / or moonju egg extract.

The external preparation composition for skin of the present invention may be applied to a cosmetic composition, a pharmaceutical composition or a soap composition.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

<Examples> Preparation of Equine Placenta Extract and Moonjulan Leaf Extract

<Example 1> Preparation Example 1 of horse placenta extract by acid hydrolysis

The frozen horse placenta was thawed to remove the amnion and cord from the placenta, and only placental chorionic tissue was collected and washed several times with water to remove blood. The placental chorionic tissue was then finely chopped and mixed with 500 ml of 3M HCl per kg of chorionic tissue and acid hydrolyzed at a temperature of 11 ° C. for 20 hours. Thereafter, the filtrate was filtered with a 5 μm filter paper, and the filtrate was concentrated under reduced pressure to evaporate hydrochloric acid. 1% (w / w) of activated carbon was added thereto, stirred for 20 minutes, and filtered again with a 5 μm filter paper. NaOH was added to neutralize the pH 6.0 ~ 7.5, filtered through a Membrane filter (0.45㎛), desalted with an electric desalter and sterilized by autoclaving (120 ℃, 20 minutes) to prepare a horse placenta extract.

Example 2 Preparation Example 2 of Equine Placenta Extract by Protease

The frozen horse placenta was thawed to remove the amnion and cord from the placenta, and only placental chorionic tissue was collected and washed several times with water to remove blood. Next, the placental chorionic tissue was finely chopped, and 1 kg of chorionic tissue, 1 L of distilled water, and 5 g of protease (NOVO) were added and reacted with stirring at 55 ° C. for 2 hours in a constant temperature water bath. Then increase the temperature to stop the enzyme reaction, filter with 5㎛ filter paper, concentrate the filtrate under reduced pressure to evaporate water, add 1% (w / w) activated carbon and stir for 20 minutes, and filter with 5㎛ filter paper. It was filtered again. After filtering with a membrane filter (0.45㎛), desalination with an electric desalter and sterilized by autoclaving (120 ℃, 20 minutes) to prepare a horse placenta extract.

Example 3 Preparation Example 3 of Solvent Extract in Horse Placenta

The frozen horse placenta was thawed to remove the amnion and cord from the placenta, and only placental chorionic tissue was collected and washed several times with water to remove blood. After the blood was removed, the villi were lyophilized to form a powder, and extracted with 3% of 80% ethanol to 300 g of the placenta while stirring at room temperature for 24 hours. After extraction, the filtrate was filtered with a 5 μm filter paper, the filtrate was concentrated under reduced pressure, water was evaporated, 1% (w / w) of activated carbon was added thereto, stirred for 20 minutes, and filtered again with 5 μm filter paper. After filtering with a membrane filter (0.45㎛), desalination with an electric desalter and sterilized by autoclaving (120 ℃, 20 minutes) to prepare a horse placenta extract.

Example 4 Preparation of Solvent Extract of Moonju Egg

2L of 80% ethanol was added to 500 g of munjoran leaf pulverized, and the mixture was extracted with stirring at room temperature for 48 hours, filtered through a 5 μm filter paper, and filtered again using a Membrane filter (0.45 μm).

<Production Example> Preparation of Cosmetic Compositions Containing Equine Placenta Extract and / or Moonjuran Leaf Extract

The formulation components shown in Table 1 below were uniformly mixed in the blending amount (% by weight) shown in Table 1 to prepare a cosmetic composition comprising the horse placenta extract and / or Moonju eggs extract of the embodiment.

Component and Compounding Amount (wt%) of Cosmetic Composition division Preparation Example 1 Preparation Example 2 Preparation Example 3 Preparation Example 4 Production Example 5 Preparation Example 6 Preparation Example 7 Example 1 3.0 Example 2 3.0 Example 3 3.0 Example 4 3.0 Example 1 + 4 3.0 Example 2 + 4 3.0 Example 3 + 4 3.0 Beta Glucan 7.0 7.0 7.0 7.0 7.0 7.0 7.0 Liquid paraffin 10.0 10.0 10.0 10.0 10.0 10.0 10.0 Beeswax 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Polysorbate 60 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Sorbitan sesquioleate 2.5 2.5 2.5 2.5 2.5 2.5 2.5 Squalane 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Butylene glycol 6.0 6.0 6.0 6.0 6.0 6.0 6.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Cetearyl alcohol 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Cetearyl Glucoside 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Incense, preservative a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water to 100 to 100 to 100 to 100 to 100 to 100 to 100 Example 1 + 4 is a mixture of the same amount of the extract of Example 1 and the extract of Example 4.
Example 2 + 4 is an equivalent mixture of the extract of Example 2 and the extract of Example 4.
Example 3 + 4 is a mixture of the same amount of the extract of Example 3 and the extract of Example 4.

Experimental Example Skin Functional Experiment

Experimental Example 1 Measurement of Melanin Synthesis Inhibition Rate

Mouse melanoma cells, B16F10 cells are penicillin-streptomycin 100 units / mL and 10% fetal bovine serum (FBS) containing a DMEM medium 37 ℃ using (GIBCO, Grand Island, NY, USA), 5% CO 2 thermostat Cultured at, and subcultured once every 4 days.

B16F10 cells were adjusted to 1 × 10 ⁵ cells / mL using DMEM medium added with 10% FBS, and then inoculated into 6 well plates, and cultured 18 hours before, as shown in <Table 1>. Example extracts or mixtures thereof (100 µg / mL) were incubated for 3 days by simultaneously treating melanin-forming stimulant α-MSH (50 nM). After 3 days, the plate was removed and washed twice with PBS. The washed cells were harvested, and the cells were completely dissolved by adding 1 N NaOH and measured by ELISA at 405 nm.

The results are shown in FIG. 1. It can be seen that both the extract and the mixture of the above example inhibit melanin production, and in particular, when the mixture of the horse placenta extract and moonjuran extract was treated, the melanin production inhibitory effect was high.

In order to confirm whether the melanin production inhibitory effect of the extract or mixture of the above example is an effect on cytotoxicity, the effect on the survival rate of B16F10 cells of the extract or the mixture of the above example was evaluated by MTT assay. B16F10 cells (2 × 10 ⁴ cells / ml) were inoculated in a 96 well plate, 18 hours before incubation, and then treated with the extract or mixture (100 μg / mL) of the above example at 37 ° C., 10% CO ₂ cells for 3 days. Cultured in the incubator. 200 μl of the MTT solution prepared at a concentration of 2 mg / ml PBS was added thereto, followed by incubation for 4 hours under the same culture conditions. The culture was removed and absorbance was measured at 570 nm by adding 200 µl of DMSO per well.

The results are shown together in FIG. 1. The results of Figure 1 show that the melanin production inhibitory effect of the extracts and mixtures of the above examples is not due to cytotoxicity.

Experimental Example 2 Skin Moisturizing Effect Experiment

The skin moisturizing effect was measured using the cosmetic composition of the preparation example and the cosmetic composition of the comparative example (the cosmetic composition of the comparative example was prepared by further comprising purified water instead of the extract or mixture content of the example).

After eighty healthy women were divided into eight groups, each composition of the preparation example and the composition of the comparative example were used on the upper part of the arm three times a day for 6 weeks, using TEWAMETER TM210 (C + K electronic GmbH. Germany). The change value (ΔTEWL) of transepidermal water loss (“TEWL”) per hour was measured. The experimental results are shown in Table 2 below.

Skin moisturizing effect Test Product △ TEWL Composition of Preparation Example 1 2.3 Composition of Preparation Example 2 2.5 Composition of Preparation Example 3 2.2 Composition of Preparation Example 4 1.7 Composition of Preparation Example 5 1.2 Composition of Preparation Example 6 1.1 Composition of Preparation Example 7 1.5 Composition of Comparative Example 4.7

The results in Table 3 show that the extracts or mixtures of the above examples all have moisturizing effects. In particular, the mixture of the extract extracts has a high moisturizing effect.

Experimental Example 3 Atopic Dermatitis Treatment Effect Experiment

Eighty patients aged 5 to 30 years with atopic dermatitis over two years were randomly selected and divided into eight groups.The cosmetic composition of each preparation and the cosmetic composition of the comparative example were compared to each group. Or a mixture of purified water instead of the mixture content), which is applied twice a morning and evening for a total of 4 weeks to the lesion site, and then the degree of improvement of atopic dermatitis as a percentage of the change in the atopic severity index (EASI score). Was measured. Here, the atopic severity index is graded into four levels of erythema, swelling, swelling, papules, scratches, and thyroiditis in the head, neck, upper extremity, torso, and lower extremities of patients with atopic dermatitis. In addition, the area score is summed to indicate the degree of skin lesions objectively, and the higher the value, the more severe atopic symptoms, and the lower, the better the skin improvement effect.

Percentage EASI score change = (earliest EASIscore after 4 weeks) / earliest EASIcore X100

The results are shown in Table 3 below.

Percentage Change in Atopic Severity Index (EASIscore) Test Product % Change in EASI score Composition of Preparation Example 1 32.5% Composition of Preparation Example 2 35.3% Composition of Preparation Example 3 29.7% Composition of Preparation Example 4 42.1% Composition of Preparation Example 5 57.3% Composition of Preparation Example 6 56.8% Composition of Preparation Example 7 52.3% Composition of Comparative Example 11.2%

Table 3 shows that the extracts and mixtures of the above examples all have atopic dermatitis improvement effects, in particular, the mixtures of the above example extracts show a high atopic dermatitis improvement effect.

Figure 1 shows the melanin production inhibitory effect and cytotoxicity evaluation results of the horse placenta extract and / or Moonjuran extract. 1, 1 is an extract of Example 1, 2 is an extract of Example 2, 3 is an extract of Example 3, 4 is an extract of Example 4, 1 + 4 is a mixture of Example 1 extract and 4 extract, 2 +4 means a mixture of Example 2 and 4 extracts and 3 + 4 means a mixture of Example 3 and 4 extracts.

Claims (12)

Includes both horse placenta extract and moonju egg extract as an active ingredient, The horse placenta extract is one of the following (I) to (III), (I) extract obtained by acid hydrolysis and neutralization of horse placental chorionic tissue from which amniotic membrane and cord were removed from horse placenta; (II) extract obtained by hydrolysis of horse placental chorionic tissue from which amniotic membrane and umbilical cord were removed from horse placenta with protease; And (III) Extract obtained by immersing the horse's placenta chorionic tissue from the amnion and umbilical cord in the placenta of water, ethanol or a mixed solvent thereof The moonjuran leaf extract is an external composition for skin whitening, characterized in that the extract obtained by immersing the moonjuran leaves in a mixed solvent of water and ethanol. delete delete Includes both horse placenta extract and moonju egg extract as an active ingredient, The horse placenta extract is one of the following (I) to (III), (I) extract obtained by acid hydrolysis and neutralization of horse placental chorionic tissue from which amniotic membrane and cord were removed from horse placenta; (II) extract obtained by hydrolysis of horse placental chorionic tissue from which amniotic membrane and umbilical cord were removed from horse placenta with protease; And (III) Extract obtained by immersing the horse's placenta chorionic tissue from the amnion and umbilical cord in the placenta of water, ethanol or a mixed solvent thereof The moonjuran leaf extract is an external composition for skin moisturizing, characterized in that the extract obtained by immersing the moonjuran leaves in a mixed solvent of water and ethanol. delete delete Includes both horse placenta extract and moonju egg extract as an active ingredient, The horse placenta extract is one of the following (I) to (III), (I) extract obtained by acid hydrolysis and neutralization of horse placental chorionic tissue from which amniotic membrane and cord were removed from horse placenta; (II) extract obtained by hydrolysis of horse placental chorionic tissue from which amniotic membrane and umbilical cord were removed from horse placenta with protease; And (III) Extract obtained by immersing the horse's placenta chorionic tissue from the amnion and umbilical cord in the placenta of water, ethanol or a mixed solvent thereof The moonjuran leaf extract is a topical composition for improving atopic dermatitis, characterized in that the extract obtained by immersing the moonjuran leaves in a mixed solvent of water and ethanol. delete delete The method according to any one of claims 1, 4 and 7, The external preparation composition for external skin is a cosmetic composition, characterized in that the cosmetic composition. The method according to any one of claims 1, 4 and 7, The external preparation composition for external skin is a soap composition, characterized in that the composition. The method according to any one of claims 1, 4 and 7, The external preparation composition for external skin, characterized in that the pharmaceutical composition.

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